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Joint Congress for

International Conference for Microcirculation and

the 13th Annual Conference of the Professional Committee for

Microcirculaion, Chinese Association of Integrative Medicine

(ICM-PCMCAIM)

(September 6-8, 2013, Beijing, China)

Organizer

The Professional Committee for Microcirculation, Chinese Association of Integrative

Medicine

Undertakers

Department of Integration of Chinese and Western Medicine/Tasly Microcirculation

Research Center, Peking University Health Science Center

President of ICM-PCMCAIM

Jing-Yan Han M.D., Ph.D.

Professor and Chairman, Department of Integration of Chinese and Western Medicine

/Tasly Microcirculation Research Center, Peking University Health Science Center

第八届全国中西医结合微循环学术会议 2008 年北京大会主席：韩晶岩

第九届全国中西医结合微循环学术会议 2009 年杭州执行主席：吕圭源

第十届全国中西医结合微循环学术会议 2010 年厦门执行主席：管昌益

第十一届全国中西医结合微循环学术会议 2011 年烟台执行主席：王世军

第十二届全国中西医结合微循环学术会议 2012 年南京执行主席：蔡辉

微循环国际研讨会暨第十三届全国中西医结合微循环学术会议

2013 年北京大会主席：韩晶岩

The 8th Annual Conference of the Professional Committee for Microcirculaion, Chinese Association of Integrative Medicine

2008, Beijing, Jing-Yan Han


2009, Hangzhou, Gui-Yuan Lv


2010, Xiamen, Chang-Yi Guan


2011, Yantai, Shi-Jun Wang


2012, Nanjing, Hui Cai

Joint Congress for International Conference for Microcirculation and the 13th Annual Conference of the Professional Committee for Microcirculaion, Chinese Association of Integrative Medicine 2013, Beijing, Jing-Yan Han

International Scientific Committee of ICM-PCMCAIM

Qi-Min Zhan, Professor and Vice-President, Chinese Academy of

Medical Sciences and Peking Union Medical College

Jing-Yan Han, Professor and Chair, Department of Integration of Chinese

and Western Medicine, Peking University Health Science

Center

Qiao-Bin Huang, Department of Pathophysiology, Southern Medical

University

Sarah Yuan, Professor and Chair, Department of Molecular Pharmacology

& Physiology, University of South Florida Morsani

College of Medicine

Makoto Suematsu, Professor and Dean, School of Medicine, Keio

University

Masato Yasui, Professor and Chair, Department of Pharmacology, School

of Medicine, Keio University

Welcome address Dear Colleagues and Friends,

On behalf of the International Scientific Committee, I am very pleased to

welcome you coming to Peking University Health Science Center for the Joint Congress of the International Conference for Microcirculation and the 13th Annual Conference of the Professional Committee for Microcirculation, Chinese Association of Integrative Medicine (ICM-PCMCAIM).

Microcirculatory disorders related diseases, such as cardiovascular disease, ischemic stroke, diabetic vascular complications and shock, have became a major impact on public health and financial expenditure worldwide. As one of the essential parts of Chinese medicine theory, the theory of qi-blood-body fluid has been effective in dealing with the problems encountered in the treatment of microcirculatory disorders clinically. However, the theory qi-blood-body fluid has not been interpreted in terms of contemporary biology.

The congress is scheduled to have plenary lectures, symposiums and poster presentations, in these sessions we are going to present and discuss the results generated from studies on the actions and mechanisms of Chinese medicine in the prevention and theraputical treatment of cardio- and cerebrovascular diseases, infection, autoimmune diseases and metabolism syndromes. In particular, we will explore the scientific basis of the theory qi-blood-body fluid with emphasis on the correlation between the theory and microcirculation. I believe these activities will certainly contribute to the understanding and cooperation between scientists and researchers in the field of microcirculation from different countries.

Autumn is the best season in Beijing. I wish you, our dear guests from United States, Japan, Britain, Switzerland, Hong Kong and other cities of China, have a merry weekend in Beijing. President of ICM-PCMCAIM

Jing-Yan Han M.D., Ph.D.

President of PCMCAIM

Professor and Chairman Department of Integration of Chinese and Western Medicine/Tasly Microcirculation Research

Center, Peking University Health Science Center

北京大学医学部西门 Peking University

Health Science Center

首都机场Capital Airport

北京西站北京站

北京南站

交通图

Traffic

会场、宾馆 Venue and Hotel

Peking University Health Science Center

YiFu Building

Master Inn

Schedule

6th Sep Friday

Room 414 Yifu Building, campus of Peking University Health Science Center

9:00-17:00 Registration 4 F, Yifu Building, campus of Peking University Health Science Center

Vision Hotel 1th F Master Inn 1th F

14:00-14:05 Opening Speech 14:05- 16:10

Symposium 1 Permeability Chairs: Jerome W. Breslin

University of South Florida Masato Yasui

School of Medicine, Keio University

S1-1 Jerome W. Breslin

University of South Florida S1-2

Jian-Gang Shen University of Hong Kong

S1-3 Qiao-Bing Huang

Southern Medical University S1-4

Chun-Shui Pan Peking University

S1-5 Masato Yasui


S1-6 Ying-Chun Yu


16:10-16:20 Tea Break 16:20-17:00 Plenary Lecture 1

The Autodigestion Hypothesis for Acute Shock: Digestive Enzymes Lurking in the Microcirculation

Speaker: Erik Bruce Kistler University of California, San Diego

Chair: Qiao-Bing Huang Southern Medical University

17:30- 20:00

Opening Reception Master Inn 2th F

20:30- 21:30

Council Meeting Room 114 Yifu Building, campus of Peking University Health Science Center

7th Sep Saturday



8:30- 9:50

Symposium 2 Metabolic cardiovascular injury and microcirculation Chairs: Pierre Maechler

University of Geneva Fu-Long Liao

China Academy of TraditionalChinese Medicine

S2-1 Fei Ye

Chinese Academy of Medical Sciences

S2-2 Li Yan


S2-3 Hai-Bo Zhu

Chinese Academy of Medical Sciences

S2-4 Yan Zhu Tianjingi University of TCM

Symposium 3 Organ injury and microcirculation Chairs: Mack Wu

University of South Florida Qiao-Bing Huang


Yu-Ling Ma University of Oxford

S3-2 Dan Zhu

Huazhong University of Science & Technology

S3-3 Xiao-Yuan Yang


S3-4 Bao-Liang Sun Taishan Medical College

9:50- 10:00

Tea Break

10:00- 10:40

Plenary Lecture 2 AQP and Brain Edema Speaker: Masato Yasui

School of Medicine, Keio

University Chair: Bao-Xue Yang

Peking University 10:40- 11:20

Plenary Lecture 3 Microvascular Permeability in Inflammation Speaker: Sarah Yuan

University of South Florida Morsani College of Medicine

Chair: Chang-Man Zhou Peking University Health Science Center

11:20 12:00

Plenary Lecture 4 Gas Biology and Medicine: How Tiny Molecules Transfer Signals Speaker: Suematsu Makoto


Chair: Xiao-Jie Xu Peking University

12:05- 12:10

Group photo Out of 1th floor Yifu Building

12:15- 13:20

Lunch Master Inn F2

13:20- 14:40

Posters P1, P2, P3, P4, P5, P6, P7, P8

14:40- 16:00

Symposium 4 Oxidative stress and microcirculation Chairs:

David Zawieja,

Texas A & M University.

Makoto Suematsu, School of Medicine, Keio University

S4-1 Xiao-Hua Guo


Rui Tan Southwest of Jiaotong University

S4-3

Symposium 5 Inflammation and microcirculation Chairs: Sarah Yuan

University of South Florida Xue-Jun Li


S5-1

Hui Cai School of Medicine, Second Military University

S5-2 Kai Sun


S5-3 Ning Yang

Chang-Man Zhou Peking University Health

Science Center S4-4

Xue-Jun Li Peking University Health Science Center

The Institute of Quality Control of Medical Material

S5-4 Hong-Quan Zhang


16:00 16:10

Tea Break

16:10- 16:50

Plenary Lecture 5 The Role and Mechanisms of Tonifing Qi and Activating Blood in Treatment for Qi Deficiency and Blood Stasis Speaker: Jing-Yan Han


Chair: Sarah Yuan University of South Florida Morsani College of Medicine

16:50- 17:30

Plenary Lecture 6 Functional Interactions of the Lymphatic and Immune Systems Speaker: David Zawieja

Texas A&M University System Health Science Center

Chair: Suematsu Makoto School of Medicine, Keio University

18:00- 20:00

Dinner Master Inn F2

Schedule 8th Sep Sunday



8:30- 9:50

Symposium 6 Thrombus Chairs: Ke-Sheng Dai

Soochow University Jian-Bo Wu Luzhou Medical College

S6-1 Ke-Sheng Dai

Soochow University S6-2

Jian-Bo Wu Luzhou Medical College

S6-3 Quan Li

Peking University S6-4

Xun-Bin Wei

Shanghai Jiao Tong University

9:50- 10:10

Tea Break

10:10- 10:50

Plenary Lecture 7 Nitric oxide: fundamental endothelial signal in vascular permeability and systemic blood pressure Speaker: Walter N. Duran

New Jersey Medical School; Rutgers, The State University of New Jersey

Chair: Jun -Bao Du Peking University Health Science Center

10:50- 11:30

Plenary Lecture 8 Mitochondria as sensor and generator of signals for the control of metabolic homeostasis Speaker: Pierre Maechler

University of Geneva. Chair: Jing-Yan Han


11:30- 12:10

Plenary Lecture 9 β-Adrenergic Receptor Subtype Signaling in the Heart: From Bench to the Bedside Speaker: Anthony Yiu Ho Woo

Peking University Chair: Hong-Quan Zhang


12:10- 12:20

Closing Ceremony

12:30- 13:30

Lunch Master Inn F2

Notice for Attendees 1. Date: September 6 (Friday) - 8 (Sunday), 2013 2. Venue: Room 414 and 402, Yi Fu Teaching Building, Peking University Health

Science Center 3. Registration 1) September 6 (Friday), 9:00-17:00 Overseas attendees: Vision hotel, First floor Lobby (No.39, Xueyuan Road,

Haidian District, Beijing. Phone: 62308899) Domestic attendees: Master Inn, First floor Lobby (No.38, Xueyuan Road, Haidian District, Beijing. Phone: 010-82320101) 2) September 6 (Friday), 13:00-17:00

September 7 (Saturday), 7:50-17:00 September 8 (Sunday ), 7:50-12:00 in front of room 414 and 402 Yi Fu Teaching Building, Peking University Health Science Center, No.38, Xueyuan Road, Haidian District, Beijing. Phone: 15901185388 (Mei-Ying Qu)

4. Conference fee 1) Domestic attendees: 1200 RMB

Overseas attendees: 200 US$ Conference fee includes Conference Guide, Proceedings, Tea break, Welcome Reception, Lunches and Dinner.

2) Students: 500 RMB 5. Hotel (at your own expense)

1) Vision hotel No.39, Xueyuan Road, Haidian District, Beijing. Phone: 0086-10-62308899 Rooms: Standard Room, Twin

One person: RMB 568/ night (1 breakfast included, RMB 70) Two person: RMB 319/ night /person (1 breakfast included,

RMB 70) Deluxe Standard, Twin

One person: RMB 728/ night (1 breakfast included, RMB 70) Two person: RMB 399/ night /person (1 breakfast included,

RMB 70) 2) Master Inn

No.38, Xueyuan Road, Haidian District, Beijing. Phone: 0086-10-82320101 Rooms: Standard Room,

One person: RMB 268/ night (breakfast included) Deluxe Standard, Twin

One person: RMB 388/ night (breakfast included) Two person: RMB 194/ night / person (breakfast included)

6. Repast Opening Reception: September 6 (Friday), 17:30-20:00, Master Inn (F2)

Dinner: September 7 (Saturday), beginning at 18:00, Master Inn (F2) Breakfast: in each of the hotel accommodation, beginning at 7:00 am (with

room card) Lunch: beginning at 12:00, Master Inn (F2)

7. Transport A. Airport to Master Inn or Vision hotel

By taxi, about RMB 110 B. Beijing west railway station to Master Inn hotel or Vision hotel

By taxi, about RMB 40 By bus, bus No.47 to “xueyuanqiao”, the north gate of Peking University Health Science Center is on the opposite side of the road.

C. Beijing south railway station to Master Inn or Vision hotel

By taxi, about RMB 80

By subway line No.4 to “haidianhuangzhuang”, change line No.10 to

“xitucheng”, Peking University Health Science Center is about 400 m to the

north, while Vision hotel is about 100 m to the north.

8. Q&A For people who would like to ask questions, please move to the venue

microphone first, and then concisely ask questions after the approval of the moderator who informs your name and affiliation to the audience.

Notice for speakers

The conference language is English. 1. Plenary Lecture: 35 min for presentation, 5 min for discussion.

2. Symposium: 15 min for presentation, 5 min for discussion

3. PC and projector are available in the conference hall. Speakers please hand over

your Power Point (2003, 2007) or U-disk, mobile hard disk or notebook computer

to the conference secretariat 60 min before your presentation. Secretariat is in

front of the conference hall, Yifu Building F5, who makes sure that your data

will not be passed to others and deletes all your data after your report.

4. Each speaker please sat at the seat designated for the next speaker in the 1st row on

the left when the one before you starts presenting. Please comply strictly the time.

5. Poster: The posters should be prepared in English with a size of 90 (width) by 120

(height) cm. Please affix poster on your assignated number before 13:30 pm on

September 6 in the poster area on the 4th floor of Yi Fu Teaching Building of the

Health Science Center Peking University. Poster communication time:

13:20-14:20, September 7. Six posters as a group, under the auspices of the host,

each poster having reporting time of 6 minutes, and 4 minutes for discussion.

Please withdraw your poster before 12:30 on September 8.

Notice for the host

1. Please seat on the seat labeled as “the next host” on the right side of the first row of

the hall before the last presentation is over.

2. The host of plenary lecture and educational lecture can make a brief remark to

introduce the speakers, according to the speakers' CV provided by the committee,

and organize a discussion after the report. Please pay attention to control time.

3. Host of symposium should introduce the speakers' names, affiliations and the titles

of the reports only, and organize a discussion after the report within the time

scheduled.

4. From 13:20 to 14:20 on September 7, host of the poster should organize the

communications in front of the appointed poster. Each paper should be reported for

6 minutes, then discussed for 4 minutes. And the host should hand in the best paper

(only one) you chose to the secretariat poster group located at the 4th floor in the

Yifu Building of Peking University Health Science Center before 17:00, September

7, including the title of the paper, the author's name and affiliation.

优秀论文奖

1. 本届会议设优秀论文奖。

2. 优秀论文奖从壁报中产生。

3. 优秀论文奖的奖状和奖金，在 9 月 8 日 12:10 的闭幕式上颁发。未到场的获

奖者将视为放弃奖项。

Award

1. The award will be set up in this conference

2. The award will be selected among the posters

3. The certificate and prize will be issued in closing ceremony at 12:10 pm in

September 8.

大会秘书处人员及联系办法：

会务负责人王传社 13911003861

报到及住宿李娜 13810060510

曲玫颖 15901185388

财务曲玫颖 15901185388

壁报阎丽 18210336659

展区王明暇 15810641735

414 会场李泉 13717600907

402 会场孙凯 13683056412

Programme

September 6 Friday Room 414 Yifu Building Campus of Peking University Health Science Center

14:05-16:10 Symposium 1

Permeability

Chairs: Jerome W. Breslin

Department of Molecular Pharmacology and Physiology Morsani College of Medicine, University of South Florida Masato Yasui

Department of Pharmacology, School of Medicine, Keio University 14:05-14:30 Thrombin and sphingosine-1-phosphate alter local lamellipodia

dynamics to modulate endothelial barrier function Jerome W. Breslin Department of Molecular Pharmacology and Physiology University of South Florida, Morsani College of Medicine

14:30-14:50 Nitric oxide/Caveolin-1/MMP pathway: a novel therapeutic strategy for

drug discovery from herbal medicine targeting blood-brain-barrier

disruption during cerebral ischemia-reperfusion injury

Jian-Gang Shen

School of Chinese Medicine, University of Hong Kong

14:50-15:10 The effects of sphingosine 1-phosphate on modulation of endothelial barrier function Qiao-Bing Huang Department of Pathophysiology, Key Lab for Shock and Microcirculation Research, Southern Medical University

15:10-15:30 Salvianolic acid B binds to Src and ameliorates hyperpermeability of

mesenteric venules in rats with endotoxmia

Chun-Shui Pan

Tasly Microcirculation Research Center, Peking University Health

Science Center

15:30-15:50 Analysis of aquaporin-mediated diffusional water permeability by

coherent anti-stokes raman scattering microscopy

Masato Yasui

Department of Pharmacology, Keio School of Medicine

15:50-16:10 CARS microscope: a novel approach to observe epithelial water

movement

Ying-Chun Yu

Department of Pharmacology, Keio University School of Medicine

16:10-16:20 Break

16:20-17:00 Plenary Lecture 1

The Autodigestion Hypothesis for Acute Shock:

Digestive Enzymes Lurking in the Microcirculation

Erik Kistler, M.D., Ph.D.

Assistant Professor, Department of Bioengineering, The Institute

of Engineering in Medicine University of California San Diego

Chair: Qiao-Bing Huang, M.D., Ph.D.

Department of Pathophysiology, Key Lab for Shock and

Microcirculation Research, Southern Medical University

17:30-20:00 Opening Reception Master Inn 2th Floor

20:30-21:30 Council Meeting Room 414

Yifu Building, campus of Peking University Health Science Center

September 7 Saturday Room 414 Yifu Building

Campus of Peking University Health Science Center

8:30-9:50 Symposium 2

Metabolic Cardiovascular injury and microcirculation

Chairs: Pierre Maechler

Department of Cell Physiology and Metabolism,

University of Geneva

Fu-Long Liao

Institute of Chinese Materia Medica, China Academy of

Traditional Chinese Medicine

8:30-8:50 A novel compound CX08005, targeting on PTP1B, improves hepatic

microcirculation disturbance, insulin resistance and

hypercholesteremia in pre-diabetic mice

Fei Ye

Beijing Key Laboratory of New Drug Mechanism of Action and

Pharmacodynamic Evaluation, Chinese Academy of Medical Sciences,

Beijing Union Medical College, Institute of Meteria Medica

8:50-9:10 Notoginsenoside R1 protects heart from ischemia reperfusion injury

via energy regulation mechanism

Li Yan


Science Center

9:10-9:30 A 1H NMR-based metabonomic investigation of time-related

metabolic trajectories of the plasma, urine and liver extracts of

hyperlipidemic hamsters

Hai-Bo Zhu

State Key Laboratory of Bioactive Substance and Function of Natural

Medicines & Ministry of Health Key Laboratory of Biosynthesis of

Natural Products, Chinese Academy of Medical Sciences and Peking

Union Medical College

9:30-9:50 Danhong injection accelerates vascular injury repair by restoring

microcirculation and promoting endothelial progenitor cell-mediated

angiogenesis.

Yan Zhu

Tianjin State Key Laboratory of Modern Chinese Medicine Tianjin University of Traditional Chinese Medicine

9:50-10:00 Break


AQP4 and Brain Edema

Masato Yasui, M.D., Ph.D.

Professor and Chair, Department of Pharmacology,


Chair: Bao-Xue Yang, M.D., Ph.D.

Professor and Vice Chair, Department of Pharmacology,

School of Basic Medical Sciences, Peking University


Microvascular Permeability in Inflammation

Sarah Yuan, M.D., Ph.D.

Professor and Chair, Department of Molecular Pharmacology and

Physiology, University of South Florida Morsani College of

Medicine

Chair: Chang-Man Zhou, M.D., Ph.D.

Professor, Department of Anatomy, Peking University Health

Sciences Center


Gas Biology and Medicine: How Tiny Molecules Transfer Signals?

Makoto Suematsu, M.D., Ph.D. Professor and Dean, School of Medicine, Keio University

Chair: Xiao-Jie Xu, Ph.D. Professor, College of Chemistry and Molecular Engineering, Peking University

12:05-12:10 Group photo

12:15-13:20 Lunch Master Inn F2

13:20-14:40 Posters

14:40-16:00 Symposium 4

Oxidative stress and microcirculation

Chairs: David Zawieja,

Department of Medical Physiology

Division of Lymphatic Biology

College of Medicine - Texas A&M Health Science Center

Makoto Suematsu,

Department of Biochemistry and Integrative Medical

Biology, School of Medicine, Keio University

14:40-15:00 ERM protein moesin is phosphorylated by advanced glycation end

products and modulates endothelial permeability

Xiao-Hua Guo

Department of Pathophysiology, Key Lab for Shock and

Microcirculation Research, Southern Medical University

15:00-15:20 Establishment of high-throughput screening for active pharmaceutical

ingredients of acute cerebral ischemia and related applied research

Rui Tan

School of Life Science and Technology, Southwest of Jiaotong

University

15:20-15:40 The symphonic mechanism of survival or death of the neurons

following ischemia stroke

Chang-Man Zhou

Department of Anatomy, Peking University Health Sciences Center 15:40-16:00 Curcumin induces autophagy to protect vascular endothelial cell

survival from oxidative stress damage

Xue-Jun Li

Department of Pharmacology, School of Basic Medical Sciences,

Peking University

16:00-16:10 Tea Break


The Role and Mechanisms of zztonifing Qi and Activating Blood in

Treatment for Qi Deficiency and Blood Stasis

Jing-Yan Han, M.D., Ph.D.

Professor and Chair, Department of Integration of Chinese and

Western Medicine/Tasly Microcirculation Research Center,


Chair: Sarah Yuan, M.D., Ph.D.


Physiology, University of South Florida Morsani College of

Medicine


Functional Interactions of the Lymphatic and Immune Systems

David C. Zawieja, Ph.D.

Regents Professor and Vice Chair, Department of Medical

Physiology

Director, Division of Lymphatic Biology College of Medicine -

Texas A&M Health Science Center

Chair: Suematsu Makoto, M.D., Ph.D.

Professor and Chair, Department of Biochemistry and Integrative

Medical Biology, School of Medicine, Keio University

18:00-20:00 Dinner

September 7 Saturday Room 402 Yifu Building



Organ injury and microcirculation

Chairs: Mack Wu Department of Molecular Pharmacology and Physiology, University of

South Florida Morsani College of Medicine

Qiao-Bing Huang Department of Pathophysiology, Key Lab for Shock and Microcirculation

Research, Southern Medical University

8:30-8:50 Pharmacological studies of mudanpi and its main active component on

cardiac ischemia and ion channels

Yu-ling Ma

Department of Physiology, Anatomy and Genetics, University of

Oxford

8:50-9:10 Tissue optical clearing for high resolution blood flow monitoring

with laser speckle contrast imaging

Dan Zhu

Britton Chance Center for Biomedical Photonics, Wuhan National

Laboratory for Optoelectronics

9:10-9:30 3, 4-dihydroxyl-phenyl lactic acid restores NADH dehydrogenase

[ubiquinone] 1 alpha subcomplex subunit 10 expression ameliorating

cardiac reperfusion injury

Xiao-Yuan Yang


Science Center

9:30-9:50 Lymphatic drainage from CNS and cerebral vasospasm following

subarachnoid hemorrhage

Bao-liang Sun

Key Lab of Cerebral Microcirculation in Universities of Shandong

(Taishan Medical University); Department of Neurology, Affiliated

Hospital of Taishan Medical University

14:40-16:00 Symposium 5

Inflammation and microcirculation

Chairs: Sarah Yuan

Department of Molecular Pharmacology and Physiology, University of South Florida Morsani College of Medicine

Xue-Jun Li Department of Pharmacology, School of Basic Medical Sciences,

Peking University

14:40-15:00 Sinomenine decreases MyD88 expression and improves inflammation-induced joint damage progression and symptoms in rat

adjuvant-induced arthritis

Hui Cai

Department of integrated traditional and western medicine, School of

Medicine, Second Military Medical University (Shanghai), Jinling

Hospital

15:00-15:20 Ginsenoside Rb1 ameliorates caveolae-mediated albumin leakage from

rat mesenteric venules induced by lipopolysaccharide

Kai Sun


Science Center

15:20-15:40 Andrographolide pills® attenuates lipopolysaccharide-Induced

pulmonary microcirculatory disturbance and acute lung injury in rats

Ning Yang

The Institute of Quality Control of Medical Material and Equipment

under the Joint Logistic Department of Beijing Military Command

15:40-16:00 Role of an integrin-interacting protein in the control of EMT in cancer

and fibrosis

Hong-Quan Zhang

Laboratory of Molecular Cell Biology and Tumor Biology, Department

of Anatomy, Histology and Embryology, School of Basic Medical

Sciences, Peking University Health Science Center

September 8 Sunday Room 414 Yifu Building



Thrombus

Chairs: Ke-Sheng Dai

Jiangsu Institute of Hematology

The First Affiliated Hospital of Soochow University

Key Laboratory of Thrombosis and Hemostasis, Ministry of

Health Suzhou

Jian-Bo Wu

Drug Discovery Research Center, Luzhou Medical College

8:30-8:50 The interaction of GPIb-IX with VWF incurs platelet activation,

apoptosis, and GPIbα ectodomain shedding

Ke-Sheng Dai

Jiangsu Institute of Hematology, The First Affiliated Hospital of

Soochow University

Key Laboratory of Thrombosis and Hemostasis, Ministry of Health

Suzhou

8:50-9:10 Targeting plasminogen activator inhibitor-1 to prevent vascular

disease

Jian-Bo Wu


9:10-9:30 Caffeic acid inhibits platelet activation and thrombus formation

Quan Li Tasly Microcirculation Research Center, Peking University Health Science

Center

9:30-9:50 Trapping red blood cells within microvessels in living mice by optical

tweezers

Xun-Bin Wei

School of Biomedical Engineering, Shanghai Jiao Tong University

9:50-10:10 Tea Break


Nitric Oxide: Fundamental Endothelial Signal in Vascular

Permeability and Systemic Blood Pressure.

Walter N. Duran, Ph.D.

Professor and Vice Chair, Department of Pharmacology &

Physiology

Professor of Surgery, Division of Vascular Surgery

New Jersey Medical School Rutgers, The State University of

New Jersey

Chair: Jun-Bao Du, M.D., Ph.D

Professor, Department of Pediatrics, Peking University First

Hospital


Mitochondria as Sensor and Generator of Signals for the Control

of Metabolic Homeostasis

Pierre Maechler, Ph.D.

Professor, Department of Cell Physiology and Metabolism,


Chair: Jing-Yan Han, M.D., Ph.D.

Professor and Chairman, Department of Integration of Chinese

and Western Medicin/Tasly Microcirculation Research Center,



β-Adrenergic Receptor Subtype Signaling in the Heart: From

Bench to the Bedside

Anthony Yiu Ho Woo, Ph.D.

Institute of Molecular Medicine, Peking University

Chair: Hong-Quan Zhang, M.D., Ph.D.


12:10-12:20 Closing Ceremony

12:30-13:30 Lunch Master Inn F2

Posters September 7 Saturday 4th Floor Yifu Building Campus of Peking University Health Science Center

Poster 1 Myocardial damage and microcirculation Chair: Chun-Shui Pan

Tasly Microcirculation Research Center, Peking University Health Science Center

13:20-13:30 P1-1 Effect of inoculated Walker256 sarcoma on angiogenesis of myocardial tissue in rats with acute myocardial infarction Jian-Gang Liu Department of Cardiology, Xiyuan Hospital, China Academy of Chinese Medical Sciences 13:30-13:40 P1-2 The ameliorating effects of long term electroacupuncture at BaiHui (DU20) and ZuSanLi (ST36) on blood pressure and myocardial hypertrophy in spontaneous hypertension rat and underlying mechanisms Ze-Jun Huo School of Acupuncture and Moxibustion, Beijing University of Chinese Medicine 13:40-13:50 P1-3 The impact of Rho kinase inhibition on cardiac and plasma angiotensin (1-7) in pressure-overload rats Hui Cai Department of Integrated Traditional and Western Medicine, Nanjing General Hospital of Nanjing Military Region PLA 13:50-14:00 P1-4 Effect of panax quinquefolius saponin on protein expression of ischemic myocardial osteopontin and tenascin-C in rats after myocardial Infarction Lei Zhang Cardiology Department of Xiyuan Hospital, China Academy of Chinese Medical Sciences 14:00-14:10 P1-5 Sang-qi granula reduces blood pressure and myocardial fibrosis by suppressing inflammatory responses associated with the peroxisome proliferator-activated receptors and nuclear factor κB protein in spontaneously hypertensive rats Lan-Yu Chen

China-Japan Friendship Hospital 14:10-14:20 P1-6 QiShenYiQi Pills@, a compound chinese medicine, ameliorates doxorubicin-induced myocardial structure damage and cardiac dysfunction in rats Dong-Xin Tang Key laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Integrated Traditional and Western Medicine, Peking University Cancer Hospital & Institute

Poster 2 Pharmacology of Chinese medicine for microcirculatory disturbance

Chair: Fei Ye

Beijing Key Laboratory of New Drug Mechanism of Action and

Pharmacodynamic Evaluation, Chinese Academy of Medical Sciences,

Beijing Union Medical College, Institute of Meteria Medica

13:20-13:30 P2-1 QiShenYiQi Pills® prevent cardiac ischemia-reperfusion injury via energy modulation Se-Qi Lin Key Laboratory of Modern Preparation of Traditional Chinese Medicine, Ministry of Education, Jiangxi University of Traditional Chinese Medicine 13:30-13:40 P2-2 Effects of Guanxinkang on blood lipids, blood rheology, atheromatous plaque and related expression in atherosclerosis of apoE-/- mice He-Wei Qin Longhua Hospital Affiliated to Shanghai University of TCM, Shanghai 200032, China 13:40-13:50 P2-3 Observation about time-effect relation of extracts from ginseng notoginseng and chuanxiong on vascular endothelial cells senescence Ming Wang The Cardiology Department of Chongqing Hospital of Traditional Chinese Medicine and Chongqing the First Hospital 13:50-14:00 P2-4 HEB improves the functional damage of gastric smooth muscle in diabetic rats Zhang-Sen Hao School of Pharmacy, Hebei Medical University, Shijiazhuang, China 14:00-14:10 P2-5 Panax notogiseng saponins ameliorates rats mesenteric microcirculatory disturbance induced by ischemia and reperufusion Yu Zhang Tasly Microcirculation Research Center, Peking University Health Science Center 14:10-14:20 P2-6 The studies of Xuefuzhuyu oral liquid intervention on the SIRT1 signal transduction mechanism in ischemic cardiomyocyte apoptosis Kui-Wu Yao Guang’anmen Hospital, China Academy of Chinese Medical Sciences

Poster 3 Platelets

Chair: Yu-Ying Liu

Tasly Microcirculation Research Center, Peking University Health Science

Center

13:20-13:30 P3-1 Arsenic trioxide induces platelet apoptosis Yi-Cun Wu

Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Jiangsu Institute of Hematology 13:30-13:40 P3-2 Effect of caffeic acid on mouse cerebral venular thrombosis induced by photochemical reaction Yu Lu Department of Gynecological, Beijing Royal Integrative Medicine Hospital 13:40-13:50 P3-3 Platelet apoptosis in uremic patients Ming Li The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology 13:50-14:00 P3-4 Surface expressions of platelet glycoprotein Ibα, GpⅡb/Ⅲa, and P-Selectin are elevated in lung cancer patients Jian-Sheng Zhang The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology 14:00-14:10 P3-5 miR30c regulates plasminogen activator inhibitor-1 in platelet Mao Luo

Drug Discovery Research Center, Luzhou Medical College 14:10-14:20 P3-6 Aspirin induces platelet apoptosis Li-Li Zhao School of Biological Science and Medical Engineering, Beijing University of Aeronautics and Astronautics

Poster 4 Lymph Chair: Hui-Ling Huang Neuro-biochemical Lab

Neurological Institute of Tianjin Tianjin Neurological Hospital

13:20-13:30 P4-1 Nitric oxide is involved in the regulating of lymphatic reactivity following hemorrhagic shock Li-Min Zhang Institute of Microcirculation, Hebei North University 13:30-13:40 P4-2 Post-hemorrhagic shock mesenteric lymph is involved in cardiac dysfunction following hemorrhagic shock Hui-Bo Du Institute of Microcirculation, Hebei North University 13:40-13:50 P4-3 Role of hydrogen sulfide on post-hemorrhagic shock mesenteric lymph drainage alleviating multiple organ injury in rats Bo Han Institute of Microcirculation, Hebei North University 13:50-14:00 P4-4 Role of myosin light chain kinase on post-hemorrhagic shock mesenteric lymph blockade ameliorating the vascular reactivity and calcium sensitivity in rats following hemorrhagic shock Yu-Ping Zhang Institute of Microcirculation, Hebei North University

14:00-14:10 P4-5 Kudiezi injection adjust PKC δ/MARCKS signal pathway and protect vascular endothelial cells after hypoxia injury Fu-Qin Chen Department of Traditional Chinese Medicine, Peking University People’s Hospital

Poster 5 Ischemia-Reperfusion injury and microcirculation

Chair: Osamu Furukawa Tasly Microcirculation Research Center, Peking University Health Science Center

13:20-13:30 P5-1 Inhibition of NADPH oxidase mediates protective effect of cardiotonic pills® against rat heart ischemia/reperfusion injury Xiao-Yuan Yang Tasly Microcirculation Research Center, Peking University Health Science Center 13:30-13:40 P5-2 Protective effects of Notoginsenoside R1 on intestinal ischemia/reperfusion injury in rats Chong Li Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University 13:40-13:50 P5-3 Inhibition of PAI-1 rescues the defect in ischemia-induced neovasculature in type II diabetic mice Rong Li

Drug Discovery Research Center, Luzhou Medical College 13:50-14:00 P5-4 Mechanism underlying trichosanthes pericarpium injection on acute myocardial ischemia protection Qi-Tao Zhao School of Basic Medicine, Shandong University of Traditional Chinese Medicine 14:00-14:10 P5-5 Oregonin from the bark of alnus japonica improves ischemia-reperfusion induced microcirculatory disturbance in rat mesentery Nguyen Huu Tung Faculty of Pharmaceutical Sciences, Nagasaki International University 14:10-14:20 P5-6 Advanced glycation end products induce endoplasmic reticulum stress through reactive oxygen species in endothelial cells Li-Li Wu Department of Pathophysiology, Key Lab for Shock and Microcirculation Research, Southern Medical University

Poster 6 Shock and inflammation

Chair: Chun-Yu Niu

Institute of Microcirculation, Hebei North University

13:20-13:30 P6-1 The effects of several vasoactive agent on intestines mesentery microcirculation in haemorrhagic shock Shang-Bin Yu Center Laboratory of Functional Science, Tongji Medical College, Huazhong University of Science and Technolgy 13:30-13:40 P6-2 Berberine inhibits Chlamydia pneumoniae infection-induced VSMC migration through downregulating expression of MMP3 and MMP9 via PI3K Jun-Yan Wei Department of Pathophysiology, School of Basic Medical Sciences, Tianjin Medical University 13:40-13:50 P6-3 Determination of sex hormone in the chronic myeloid leukemia patients treated with TKI Xiao-Hui Chang The 210th Hospital of PLA, TCM Hematology Center 13:50-14:00 P6-4 Epidemical survey of PLGC of phlegm-accumulation stasis and association with the polymorphism of TLR4 Gene Mu-Xin Wei

Institute fo Integreted Medicine, Nanjing Medical University 14:00-14:10 P6-5 Huang Qi Jian Zhong Pellet® repairs colonic mucosal injury induced by 2,4,6-trinitrobenzene sulfonic acid in rats via mechanisms involving improvement of energy metabolism Duan-Yong Liu Jiangxi University of Traditional Chinese Medicine 14:10-14:20 P6-6 Protective effects of paeonol on lipopolysaccharide-induced mesenteric microcirculatory disturbance in rats Lei Dong

Department of Critical Care Medicine, Beijing Friendship Hospital, Capital Medical University

Poster 7 Brain injury and microcirculation Chair: Shi-Jun Wang Shandong University of Traditional Chinese Medicine 13:20-13:30 P7-1 Long-term stimulation with electroacupuncture at DU20 and ST36 attenuates cerebral blood flow and rescues hippocampal neuron in spontaneously hypertensive rats Gui-Hua Tian

School of Acupuncture and Moxibustion, Beijing University of Chinese Medicine 13:30-13:40 P7-2 Adoptive regulatory T-cell therapy prevents cerebral vasospasm and reduces early brain injury following experimental sah Li-Ping Zhang Department of Neurology and Key Lab oratory of Cerebral Microcirculation, Taishan Medical University 13:40-13:50 P7-3 Intravenous transplantation of In vitro expanded CD4+CD25+ regulatory T cells exerts protective effects and promotes the expression of Iba1, GFAP after cerebral Ischemia-Reperfusion in rats Yan Cheng Key Lab of Cerebral Microcirculation in Universities of Shandong (Taishan Medical University); Department of Neurology, Affiliated Hospital of Taishan Medical University 13:50-14:00 P7-4 Anti-inflammation treatment after subarachnoid hemorrhage: regulatory T-cells adoptive transfer Lei Zhang Key Lab of Cerebral Microcirculation in Universities of Shandong (Taishan Medical University); Department of Neurology, Affiliated Hospital of Taishan Medical University 14:00-14:10 P7-5 Blocking neurodegenerative damages of Alzheimer's disease by aromatic glycosides in multiple pathways Ying-Hong Wang Department of Combined Traditional Chinese Medicine with Western Medicine, School of Basic Medical Science, Peking University 14:10-14:20 P7-6 The involvement of programmed cell death 5 (PDCD5) in the regulation of apoptosis in cerebral ischemia/reperfusion injury Chun-Hua Chen Department of Anatomy and Embryology, Peking University Health Science Center

Poster 8 Clinic and microcirculation Chair: Hui Cai

Department of Integrated Traditional and Western Medicine, Nanjing General Hospital of Nanjing Military Region PLA

13:20-13:30 P8-1 The correlation analysis between the visualization of hemorrheologic changes and platelet function on the patients with acute coronary syndrome after percutaneous coronary intervention Jia-Qi Wang Xiyuan Hospital, China Academy of Traditional Chinese Medicine 13:30-13:40 P8-2 To observe the effect of Tong-Mai-Yang-Xin pill in adjuvant treatment of three kinds of blood diseases Ai-Hong Wei The 210th Hospital of PLA, TCM Hematology Center 13:40-13:50 P8-3 Analysis on the thinking of using non-traditional promoting blood circulation to remove blood stasis drugs to improve the microcirculation Wan-Li Gu Department of traditional Chinese medicine, Liaocheng People's Hospital 13:50-14:00 P8-4 Study for the treatment of integrative traditional Chinese and western medicine on acute-on-chronic or subacute-on-chronic liver failure in a multiregional randomized and controlled trial Jun Li The 302 military hospital of P.L.A 14:00-14:10 P8-5 Hemorheology visualization clinical research of elderly hypertension with different TCM syndromes Xiong Wei Xiyuan Hospital, China Academy of Chinese Medical Sciences 14:10-14:20 P8-6 Rhubarb root and rhizome-based Chinese herbal prescriptions for acute ischemic stroke: a systematic review and Meta-analysis of 12 randomized controlled trials Guo-Qing Zheng The Center of Neurology and Rehabilitation, The Second Affiliated Hospital of Wenzhou Medical University

Plenary Lecture 1

16:20-17:00 September 6 Friday

Room 414 Yifu Building

The Autodigestion Hypothesis for Acute Shock:

Digestive Enzymes Lurking in the Microcirculation

Erik Kistler, M.D., Ph.D.

Assistant Professor, Department of Bioengineering, The Institute

of Engineering in Medicine, University of California San Diego

Chair : Qiao-Bing Huang, M.D., Ph.D.

Department of Pathophysiology, Key Lab for Shock and Microcirculation

Research, Southern Medical University

PL-1 The Autodigestion Hypothesis for Acute Shock: Digestive Enzymes Lurking in the Microcirculation Geert W. Schmid-Schönbein, Angelina E. Altshuler, Frank A. DeLano, Alex A. Penn, and Erik Kistler Department of Bioengineering, The Institute of Engineering in Medicine University of California San Diego, La Jolla, California 92093 – 0412, USA Physiological shock and multi-organ failure (after trauma, loss of blood volume, surgery, burns, infections for example) is a worldwide problem associated with high mortality. It is an emergency situation during which organs fail, for example by fluid accumulation in the lung, known as acute respiratory distress syndrome, a lack of urine production due to renal failure, or in form of blood pressure reduction due to cardiac and peripheral vascular failure. No effective treatment exist other than alleviation of symptoms. In spite of numerous suggestions no agreement exists for the microvascular and molecular mechanisms that lead to the relative rapid cell and organ failure during shock. We propose here a new hypothesis for the mechanisms that leads to cell dysfunction and organ failure in shock and multi-organ failure associated with the pancreatic digestive enzymes (Kistler et al., 2000). These powerful enzymes are synthesized in the pancreas and transported in the lumen of the small intestine as requirement for normal food digestion. They are fully activated and remain compartmentalized in the lumen of the intestine by the mucosal barrier. But in intestinal ischemia or after exposure to inflammatory mediators the permeability of the mucosal barrier increases and the pancreatic enzymes escape into the wall of the intestine (Chang et al., 2012). Entry of the digestive enzymes into the wall of the intestine precipitates an autodigestion process in which structures of the intestine are destroyed. The consequence is destruction of interstitial structures and cell membrane molecules, including E-cadherin between epithelial cells, thus compromising the tight intestinal barrier properties of the intestine. The pancreatic enzymes also generate breakdown products during autodigestion, of which unbound free fatty acids are especially cytotoxic. The digestive enzymes and the breakdown products they generate escape into the systemic circulation where they generate multiorgan failure. We present evidence to block the digestive enzymes in acute forms of shock as a potential therapeutic intervention (Mitsuoka et al., 2000; Fitzal et al., 2002; DeLano et al., 2013). The therapeutic value of blocking digestive enzymes in the lumen of the intestine remains to be determined in patients. References Chang, M., Kistler, E., Schmid-Schönbein, G.W.: Disruption of the intestinal mucin layer during ischemia allows early entry of digestive enzymes into the intestinal wall. Shock, 37:297-305, 2012. Fitzal, F., DeLano, F.A., Young, C., Rosário, H.S, Schmid-Schönbein, G.W.: Pancreatic protease inhibition during shock attenuates cell activation and peripheral inflammation. J. Vasc. Res., 39:320-329, 2002. Kistler, E.B., Hugli, T.E., Schmid-Schönbein, G.W.: The pancreas as a source of cardiovascular cell activating factors. Microcirculation, 7:183-192, 2000. Mitsuoka, H., Kistler, E.B., Schmid-Schönbein, G.W.: Generation of in vivo activating factors in the ischemic intestine by pancreatic enzymes. Proc. Nat. Acad. Sci. U.S.A., 97:1772-1777, 2000. DeLano, F.A., Hoyt, D.B., Schmid-Schönbein, G.W.: Digestive enzyme blockade in the lumen of the intestine increases long-term survival rate after shock. Science Translational Medicine, 5(169):169ra11, 2013.

Erik B. Kistler is Assistant Professor, Director of Intensive Care Research, in the

Department of Anesthesiology & Critical Care at the University of California, San Diego (UCSD) and Attending Anesthesiologist at the Veteran’s Administration (VA) Hospital, San Diego, California. Dr. Kistler received his Ph.D. degree in Bioengineering at UCSD under the mentorship of Professor Geert W. Schmid-Schönbein, and was a Post-doctoral Fellow in the Department of Immunology at The Scripps Research Institute in La Jolla, CA. Dr. Kistler received his M.D. degree at the George Washington University School of Medicine and Health Sciences in Washington, D.C. His internship was in the Department of Surgery at Harbor-UCLA and residency was in the Department of Anesthesiology & Critical Care at UCSD. He subsequently completed a Fellowship in Critical Care at the Massachusetts General Hospital, after which he accepted an appointment in the prestigious Professor ‘X’ series at UCSD, the only member of his Department in this track, which is designated for high performing physician-scientists in both clinical and research endeavors. Dr. Kistler also has degrees in Electrical Engineering and Music, and has worked in the aerospace industry prior to his medical education. Dr. Kistler is active in the Societies of Critical Care Medicine, Critical Care Anesthesiologists, Shock, and the American Society of Anesthesiologists, among other organizations. Dr. Kistler has received several awards for his research and has been independently funded since shortly after arriving on faculty. In addition to his full-time position as a clinician in the Intensive Care Unit and the operating theater, Dr. Kistler maintains a full-time laboratory presence and conducts clinical research in the field of critical care. His research interests are very translational, aimed at bringing bench-top therapies to the bedside and new hypotheses from the bedside back to the laboratory. Among his interests are inflammation and shock, particularly with relevance to the microcirculation, critical care outcomes optimization and goal-directed therapy. Much of Dr. Kistler’s laboratory work is in conjunction with Dr. Schmid-Schönbein, emphasizing a bioengineering analysis of the microcirculation in human disease. This effort has resulted in the discovery of a fundamental mechanism for cell dysfunction and inflammation due to “Auto-digestion”. The team proposed a previously unrecognized mechanism for Shock and Multi-organ Failure due to unchecked degrading protease activity.

Plenary Lecture 2

10:0010:40 September 7 Saturday



Masato Yasui, M.D., Ph.D.

Professor and Chair, Department of Pharmacology, School of Medicine,

Keio University

Chair：Bao-Xue Yang, M.D., Ph.D.

Professor and Vice Chair, Department of Pharmacology, School of Basic

Peking University

PL-2


Masato Yasui

Department of Pharmacology, School of Medicine, Keio University, Tokyo, JAPAN

Aquaporin-4 (AQP4) is the main water channel in the brain and is distributed with highest

density in the perivascular and subpial astrocyte end-feet. AQP4 is a critical component of an

integrated water and potassium homeostasis. Indeed, AQP4 has been implicated in several

neurologic conditions, such as brain edema, seizure and even mood disorders. Expression and

regulation of AQP4 have been studied to understand the roles of AQP4 in physiological and

pathological conditions. Molecular mechanisms how AQP4 is dynamically regulated have been

shown at different levels such as channel gating, subcellular distribution, phosphorylation,

protein-protein interactions and orthogonal array formation. We found that AQP4 is rapidly and

reversibly regulated by Zinc and propofol on a dose-dependent manner. Immunological

function of AQP4 was also demonstrated against inflammatory response in brain disease

models. Interestingly, AQP4 was identified as a target antigen of autoimmune disease,

neuromyelitis optica (NMO). We evaluated putative epitopes on AQP4 for NMO-IgG binding.

AQP4 may be a potential therapeutic target in several neurologic conditions including brain

edema. Further studies from different aspects are required to develop new drugs against AQP4.

Masato Yasui Personal Birth: June 28, 1964，Tokyo, Japan Residence: 2-2-14 Sasuke, Kamakura, Kanagawa, 248-0017 JAPAN Education Medical License (M.D.), Graduated from School of Medicine, Keio University (Tokyo, Japan) Doctor of Philosophy (Ph.D.) at the Karolinska Institute (Stockholm, Sweden). Thesis Title: Expression and Regulation of water channels and ion transporters (Ontogenic Aspects) Doctor of Medical Science (Ph.D.) at School of Medicine, Keio University (Tokyo, Japan) Research Training Guest Researcher, Dept. of Molecular Neurobiology University of Tokyo, Tokyo, Japan Postgraduate Student, Dept. of Woman and Child Health, Pediatric Unit, Karolinska Institute, Stockholm, Sweden (Prof. Anita Aperia) 1997-2000 Postdoctoral Research Fellow, Dept. of Biological Chemistry Johns Hopkins Univ. School of Medicine, Baltimore, USA (Prof. Peter Agre) Academic Positions Instructor, Dept. of Biol. Chem., Johns Hopkins School of Medicine, Baltimore, Maryland, USA 2001- Assistant Professor, Department of Pediatrics and Biological Chemistry, Johns Hopkins

School of Medicine, Baltimore, Maryland, USA Professor and Chair, Department of Pharmacology Keio University School of Medicine, Tokyo Japan

Honors Human Frontier Science Program, Long-term Fellowship Award American Heart Association MDA Fellowship Award 2000 Johns Hopkins Young Investigators Day,

A. McGehee Harvey Prize for Post-doctoral research 2002 Keio Medical School, Alumni Assoiciation Award, Tokyo

Biennial Spa Foundation Prize (shared with Peter Agre), Brussels 2004 S&R Foundation Award, Washington, D.C. Selected Publications Yasui, M., Kwon, T-H., Knepper, M. A., Nielsen, S. & Agre, P. Aquaporin-6: An intracellular vesicle water channel protein in renal epithelia. Proc. Natl. Acad. Sci. USA 96, 5808-5813 (1999). Yasui, M., Hazama, A., Kwon, T-H., Nielsen, S., Guggino, W.B. & Agre, P. Rapid gating and anion permeability of an intracellular aquaporin. Nature 402, 184-187 (1999). Liu, K., Kozono, D., Kato, Y., Agre, P., Hazama, A., and Yasui, M. Conversion of aquaporin-6 from an anion channel to a water-selective channel by a single amino acid substitution. Proc. Natl. Acad. Sci. USA 102, 2192-2197 (2005).

Plenary Lecture 3




Sarah Yuan, M.D., Ph.D.


Physiology, University of South Florida Morsani College of Medicine

Chair: Chang-Man Zhou, M.D., Ph.D.

Professor, Department of Anatomy, Peking University Health Sciences

Center

PL-3


Sarah Yuan

Department of Molecular Pharmacology and Physiology, University of South Florida Morsani

College of Medicine, Tampa, Florida 33612, USA.

The vascular endothelium lining on the inner surface of blood vessels serves as the first

interface for circulating blood components to interact with cells in the vascular wall and its

surrounding tissues. In addition to regulating blood delivery and perfusion, a major function

of vascular endothelia, especially those in exchange microvessels (capillaries and postcapillary

venules), is providing a semi-permeable barrier that controls blood-tissue exchange of fluids,

nutrients and metabolic wastes while preventing pathogens or harmful materials in the

circulation from entering into tissues. During host defense against infection or tissue injury,

endothelial barrier dysfunction occurs as a consequence as well as cause of inflammatory

responses. Plasma leakage disturbs fluid homeostasis and impairs tissue oxygenation, a

pathophysiological process contributing to multiple organ dysfunction associated with trauma,

infection, metabolic disorder, metastatic tumor development, and other forms of disease. This

lecture provides an updated overview of microvascular endothelial barrier structure and

function in health and disease. The discussion is initiated with the basic physiological principles

of fluid and solute transport across microvascular endothelium, followed by detailed

information on endothelial cell-cell and cell-matrix interactions and the experimental

techniques that are employed to measure endothelial permeability. Further discussion focuses

on the signaling and molecular mechanisms of endothelial barrier responses to various

stimulations or drugs, as well as their relevance to several common clinical conditions. This

comprehensive analysis of microvascular endothelial barrier pathophysiology would assist

research scientists as well as clinicians in advanced translational research for improved health

care.

Sarah Y. Yuan, M.D., Ph.D. Sarah Y. Yuan is Professor and Chair of the Department of Molecular Pharmacology and Physiology at the University of South Florida Morsani College of Medicine. She also holds a prestigious position as the Bob and Evelyn Deriso Endowed Chair in Cardiovascular Research and Professor of Surgery in USF. She was recently recruited from the University of California, Davis, where she served as the Pearl Stewart Professor in Surgery and Director, Division of Research. Dr. Yuan is an internationally recognized leader in microcirculation research. Her laboratory conducts translational research investigating the cellular and molecular regulation of cardiovascular function in health and disease states, including inflammation, trauma, infection, diabetes, atherosclerosis, and cancer metastasis. The lab employs multifaceted research approaches examining disease pathology in human subjects, testing physiological processes in animal models, and identifying key molecules from cultured cells. Her achievements in these areas are highlighted by a successful development of sophisticated microsurgery procedures and imaging techniques for in-depth analyses of microvascular barrier structure and function, and identification of novel signal pathways altering microvascular permeability as potential therapeutic targets for prevention or treatment of inflammatory diseases. Over the past twenty years, Dr. Yuan’s research has been continuously funded by the National Institutes of Health and has resulted in extensive publications in high impact journals. Her contributions to the scientific community worldwide are indicated by many accolades from various international societies and invitations by foreign countries to evaluate their national grants or research proposals. She was Associate Editor of Microcirculation and is currently an active member of several editorial boards. In addition, Dr. Yuan has served on multiple study sections or grant panels at the NIH, VA, and other countries. Dr. Yuan’s training background as a trauma surgeon along with her expertise in cellular and molecular biology enables her to closely interact with clinicians and basic scientists at different career stages while providing unique mentorship in translational research. Her lab has served as a training base for scholars from various programs to learn advanced experimental skills, where she has mentored numerous pre-doctoral and post-doctoral trainees across multiple disciplines. The majority of her trainees have successfully advanced in their academic careers. Currently, as the chair of a large multi-disciplinary academic department in a highly rated research university, Dr. Yuan has embraced a leadership role in the further development of cardiovascular research and educations programs toward scientific excellence and international prominence.

Plenary Lecture 4

11:30-12:10 September 7 Saturday


Gas Biology and Medicine: How Tiny Molecules Transfer Signals ?

Makoto Suematsu, M.D., Ph.D.

Professor and Dean, Department of Biochemistry and Integrative Medical


Chair: Xiao-Jie Xu, M.D., Ph.D.

Professor, College of Chemistry and Molecular Engineering,

Peking University

PL-4

Gas Biology and Medicine: How Tiny Molecules Transfer Signals ? Makoto Suematsu, M.D., Ph.D. Professor and Chair, Department of Biochemistry, School of Medicine, Keio University, JST ERATO Suematsu Gas Biology Project, Tokyo 160-8582, Japan Heme oxygenase (HO)/CO protects cancer cells against oxidative stress, while the

gas-responsive signaling mechanisms remain unknown. We have recently revealed that CO

constitutively generated in brain inhibits H2S-producing cystathionine -synthase (CBS) and

thereby regulate cerebral cortex microvessels tonically; the mechanisms contribute to hypoxic

vasodilation, since hypoxia downregulates CO acutely to activate CBS in astrocytes, causing

H2S-dependent microvascular dilatation. Since such an effect of CO on CBS might alter

methionine metabolism that resides in the upstream of the enzyme, we inquired whether

stress-inducible CO might alter protein methylation to regulate biological systems.

Metabolomics in cancer cells allowed us to show that CO-sensitive methylation of PFKFB3, an

enzyme producing fructose 2,6-bisphosphate (F-2,6-BP), serves as a switch to activate

phosphofructokinase-1, a rate-limiting glycolytic enzyme. In human leukemia U937 cells,

PFKFB3 constitutes asymmetrically di-methylated form at R131 and R134 through

modification by protein arginine methyltransferase-1. HO-1 induction or CO de-methylates

PFKFB3 in varied cancer cells to suppress F-2,6-BP, shifting glucose utilization from

glycolysis toward pentose phosphate pathway. PFKFB3 demethylation depends on inhibitory

effects of CO on heme-containing cystathionine -synthase (CBS) that modulate remethylation

metabolism, and increases NADPH to supply reduced glutathione that protects against

oxidative stress and anti-cancer reagents. Once demethylated, PFKFB3 undergoes

polyubiquitination to be degraded in proteasomes. These results suggest that

CO/CBS-dependent PFKFB3 demethylation determines directional glucose utilization to ensure

resistance against oxidative stress for cancer cell survival.

References

Morikawa T, et al., PNAS 109, 1293-1298, 2012

Nishida M, et al., Nat Chem Biol 8, 714-24, 2012

Makoto Suematsu, M.D., Ph.D.

Dean, School of Medicine, Keio University Birthday: November 30, 1957 （54 years old） Nationality: Japan Present Address: 2-35-8 Zenpukuji, Suginami-ku Tokyo 167-0041, Japan March 1983 Graduated from Keio University School of Medicine (M.D.) 1984-1988 Post-graduate School, Keio University School of Medicine (Ph.D.) （Gastroenterology and Microvascular Physiology) May 1991 Bioengineer Step IV, Institute for Biomedical Engineering,

University of California San Diego (Supervised by Professor Benjamin W Zweifach and Professor Geert W Schmid-Schoenbein)

April 2001 Professor and Chair, Department of Biochemistry and Integrative Medical Biology, Keio University School of Medicine

April 2003 Leader, National Leading Project for Biosimulation by Ministry of Education, Sciences and Technology

June 2007 Leader, Global Center of Exellence (GCOE) for Life Sciences, Human Metabolomic Systems Biology from MEXT

October 2007-present Dean, School of Medicine, Keio University October 2009-present Leader, JST, ERATO, Suematsu Gas Biology Project IV. Main Research Interests Gas Biology, Microcirculatory physiology Major publication Suematsu, M., et al. Carbon monoxide: An endogenous modulator of sinusoidal tone in the perfused rat liver. J. Clin. Invest. 96, 2431-2437, 1995. Goda, N., et al. Distribution of heme oxygenase isoforms in rat liver: Topographic basis for carbon monoxide-mediated microvascular relaxation. J. Clin. Invest. 101, 604-612, 1998. Hayashi, S., et al. Induction of heme oxygenase-1 suppresses venular leukocyte adhesion elicited by oxidative stress: Role of bilirubin generated by the enzyme. Circ. Res. 85, 663-671, 1999 Kyokane, T., et al. Carbon monoxide from heme catabolism protects against hepatobiliary dysfunction in endotoxin-treated rat liver. Gastroenterology 120, 1227-1240, 2001. Murai, M., et al. Subendothelial dome of gut Peyer’s patches is the essential anatomical site in initiating acute graft-versus-host disease. Nature Immunol. 4, 154-160, 2003. Soga, T., et al. Differential Metabolomics Reveals Ophthalmic acid as an oxidative stress biomarker indicating hepatic glutathione consumption. J. Biol. Chem. 281(24) 16768-16776, 2006. Shintani, T., et al. Cystathionine -synthase as a CO-sensitive regulator of bile excretion. Hepatology 49, 141-150, 2009. Kajimura M, et al. Interactions of multiple gas-transducing systems: Hallmarks and uncertainties of CO, NO and H2S gas biology. Antioxid. Redox Signal. 13, 157-192 (Invited review) 2010. Hattori K, et al. Paradoxical ATP elevation in ischemic penumbra revealed by quantitative imaging mass spectrometry. Antioxid Redox Signal 13, 1157-1167 (News & Views) 2010. Takubo K, et al. Regulation of the HIF-1 level is essential for hematopoietic stem cells. Cell Stem Cell 7, 391-402, 2010. Ishimoto T, et al CD44 variant regulates redox status in cancer cells by stabilizing the xCT subunit of system xc- and thereby promotes tumor growth. Cancer Cell 2011, 19(3), 387-400. Morikawa T, et al. Hypoxic regulation of the cerebral microcirculation is mediated by a carbon monoxide-sensitive hydrogen sulfide pathway. Proc Natl Acad Sci USA 2012 in press.

Plenary Lecture 5

16:10-16:50 September 7 Saturday


The Role and Mechanisms of Tonifing Qi and Activating Blood in

Treatment for Qi Deficiency and Blood Stasis


Professor and Chairman, Department of Integration of Chinese and

Western Medicine/Tasly Microcirculation Research Center, Peking

University Health Science Center

Chair: Sarah Yuan, M.D., Ph.D.


Physiology, University of South Florida Morsani College of Medicine

PL-5 The Role and Mechanisms of Tonifing Qi and Activating Blood in Treatment for Qi Deficiency and Blood Stasis Jing-Yan Han Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China Blood vessel occlusion due to thrombus, vessel contraction or atherosclerotic results in a decreased supply of the oxygen and nutritions in the perfused territory, which downregulates the expression of ATP synthase related proteins while increases ATP degradation, leading to ATP depletion.. ATP depletion not only causes cytoskeleton degradation, but also impairs cell functions, including dysfunction of myocardium contraction and vascular endothelium hyperpermeability. The dysfunction of myocardium contraction slows down the blood flow leading to blood stasis, wheres microvascular hyperpermeability allows extravasation of serum album and water which result in peri microvascular edema. After recanalization, hypoxanthine, the degradation product of ATP, may release negative oxygen ions in the presence of hypoxanthine oxidase and resupplied oxygen, water and nutrition, leading to the well known ischemia and reperfusion injury. Qi in Chinese medicine involves oxygen inhaled through lungs, nutrition absorbed from intestines, and ATP synthesized. The vessel occlusion and the resultant anoxia, lack of nutrition and depletion of ATP collectively coincide with the concept Qi deficiency. Accordingly, Qi deficiency decreases heart function and blood flow leading to microcirculatory disorders which belongs to Qi deficiency and blood stasis; while Qi deficiency caused microvascular hyperpermeability and albumin extravasation belong to qi failing to control blood. In Chinese medicine, tonifying Qi and activating blood is an efficient strategy for treatment of Qi deficiency and blood stasis, and tonifying Qi and consolidation is used to cope with Qi deficiency and failing to control. The speaker presents data showing the beneficial role of Qishenyiqi pills, a compound

Chinese medicine for tonifying Qi and activating blood, in rat myocardium injury and cardiac

microcirculatory hyperpermeability. Explored are also the mechanisms underlying the Qi

deficiency and blood stasis as well as the Chinese medicine for tonifying Qi and activating

blood. The results provide novel insights for therapy of serum albumin extravasation during

ischemia / reperfusion injury and shock.

Educations

Date and Place of Birth：1958.8.22. Shenyang


Western Medicine, School of Basic Medical Science, Peking University

Director, Tasly Microcirculation Research Center, Peking University Health


1977-1982 Liaoning University of Chinese Medicine (M.D.) 1986-1989 Graduate School of Liaoning University of Chinese Medicine (M.S.) 1993-1999 Postgraduate School, Keio University (2002 Ph.D.) Professional and Teaching Experiences 1982-1986 Assistant Professor, Department of Basic Chinese Medicine, Liaoning

University of Chinese Medicine 1989-1991 Assistant Professor, Department of Pathology and Physiology, 2nd Affiliated

Hospital, Liaoning University of Chinese Medicine 1991-1993 Visiting Researcher, Department of Internal Medicine, Keio University School

of Medicine 2000-2002 Visiting Researcher, Department of Internal Medicine, Keio University School

of Medicine 2002-2009 Co-Researcher, Department of Internal Medicine, Keio University School of

Medicine 2004-present Director, Tasly Microcirculation Research Center, Peking University Health

Science Center 2008-present Professor and Chairman, Department of Integration of Chinese and Western

Medicine, School of Basic Medical Science, Peking University 2009-present Guest Professor, Department of Internal Medicine, Keio University School of

Medicine 2010-present Dean, Department of Integration of Chinese and Western Medicine, Peking

University Health Science Center He is a President, Professional Committee for Microcirculation of Chinese Association of Integrative Medicine (PCMCAIM), Vice-President of Chinese Society for Microcirculation. Member of the International Liaison Committee for Microcirculation. He has published over 40 original articles and reviews in the last 10 years. He has demonstrated the pivotal role of microcirculatory disturbances rather than macrovascular deficits in the whole pathological course of blood stasis related diseases.

Plenary Lecture 6



Functional Interactions of the Lymphatic and Immune Systems

David C. Zawieja, Ph.D.

Regents Professor and Vice Chair, Department of Medical Physiology

Director, Division of Lymphatic Biology


Chair: Makoto Suematsu, M.D., Ph.D.

Professor and Chair, Department of Biochemistry and Integrative Medical


PL-6 Functional Interactions of the Lymphatic and Immune Systems David Zawieja Texas A&M University Health Science Center, College of Medicine, Department of Medical

Physiology and Division of Lymphatic Biology, Temple Texas, 76504, USA.

Many immune cells are classically thought to traffic through the lymphatics on their route to the

nodes as part of the normal innate and learned immune response. The architecture of the

mesenteric lymphatic network provides a unique site for entrance of macromolecules and cells

absorbed from the intestine to the lymph stream. These can be transported, via mesenteric

pre-nodal lymphatics (PreNL), to the nodes in which innate and learned immune response are

initiated. Agents that leave the node in lymph are then carried by post-nodal collecting

lymphatics (PstNL), and thoracic duct (TD) to the blood. Antigen-presenting cells (APCs) are

specialized cells that probe, recognize, phagocytose & display antigen complex with major

histocompatibility complex II (MHCII) on its surface, to start the immune response. They are

classically thought to sense antigens in the tissues, home to and enter initial lymphatics en route

to the node via lymph flow. We provide evidence of a resident population of APC in the PreNL

that are uniquely positioned to sense and present antigens directly from lymph. We measured

the distribution and morphology of the APCs in rat mesenteric PreNL, PstNL and TD by

fluorescence immunohistochemistry. We demonstrate that: 1. MHCII-positive APCs reside

within the muscularized lymphatic wall that have pseudopods that extend to the endothelium

subendothelium. 2. These APCs rapidly (<30 minutes) sense antigens within the lymph that

entered at sites far upstream. 3. Local histological microenvironments mould APCs, such that

histologically the APCs in the PreNL wall had the richest cytoplasm, longest processes, and

most irregular cellular shape with statistically highest cellular density and pseudopod number

per APC compared to PstNL and TD. These novel results indicate that muscularized lymphatics

rapidly sense antigens and may play an important role in the transition from an innate to a

learned immune response.

David C. Zawieja, Ph.D. David C. Zawieja, is a Texas A&M System Regents Professor and the Vice-Chair of the Department of Medical Physiology in the College of Medicine at the Texas A&M University Health Science Center. He also serves as the founding Director of the Division of Lymphatic Biology and the Director of the Integrated Microscopic Imaging Laboratory in the Department of Medical Physiology at TAMUHSC. He received his Bachelor of Science, degree from the University of Wisconsin - Green Bay with majors in: Biology, Chemistry, and Population Dynamics. He started his graduate career in the Graduate Program in Biomedical Engineering at Rensselaer Polytechnic Institute, Troy, NY. He obtained his Doctor of Philosophy degree in Physiology at the Medical College of Wisconsin, Milwaukee, WI in the Department of Physiology. He performed his Postdoctoral training at Texas A&M University, College of Medicine in the Department of Medical Physiology. Dr. Zawieja is recognized as an international leader in research in the microcirculation, particularly in lymphatic biology and microcirculatory exchange. His laboratories in Temple and College Station Texas, investigate the microcirculation, primarily the control and regulation of fluid and macromolecular transport throughout the three microcirculatory compartments: blood, interstitial and lymphatic. Much of his work specifically focuses on the function of the lymphatic system, defining the mechanisms responsible for the generation and regulation of lymph flow as well as delineating the roles of lymphatic function in body fluid homeostasis, lipid absorption/metabolism and immunity during health and disease. His expertise and leadership in lymphatic biology is acknowledged through his roles as the Chair of the Scientific and Medical Advisory Council of the Lymphatic Research Foundation, as a member of the Trans NIH Coordinating Committee on Lymphatic Research and the 2010 winner of the Lymphatic Research Foundation’s Lymphatic Research International Leadership Award. He has served the microcirculatory field as a long-standing member of the Executive Council of the Microcirculatory Society and the MCS President from 2010-2011. He was an Associate Editor of the journals Microcirculation and Frontiers in Vascular Physiology as well as a member of the Editorial Board of the journals Microcirculation and Lymphatic Research and Biology. He is a Fellow of the Cardiovascular Section of the American Physiological Society. He has had continuous grant support from NIH since 1987 as well as support from NASA, the National Space Biomedical Research Institute, and the American Heart Association among others. He has served as a grant reviewer for international, national and regional granting agencies including; the NIH, American Heart Association, NSF, the Lymphatic Research Foundation, American Cancer Society, Cancer Research UK among others. He has given over 100 invited lectures at meetings, conferences, symposia and departments around the world and organized and chaired numerous national and international meetings and symposia. He has published over 100 papers, reviews and book chapters as well as 200 abstracts. He has reviewed for numerous journals. He has trained over 30 graduate students as mentor/chair or graduate committee member and served as advisor for 10 postdoctoral fellows. In addition he has served as host and trainer for dozens of scientists from around the world in the fields of lymphatic biology, microcirculation and imaging.

Plenary Lecture 7

10:10-10:50 September 8 Sunday


Nitric Oxide: Fundamental Endothelial Signal in Vascular

Permeability and Systemic Blood Pressure


Professor and Vice Chair, Department of Pharmacology & Physiology

Professor of Surgery, Division of Vascular Surgery

New Jersey Medical School

Rutgers, The State University of New Jersey

Chair: Jun-Bao Du, M.D., Ph.D.

Professor, Department of Pediatrics, Peking University First Hospital

PL-7 Nitric Oxide: Fundamental Endothelial Signal in Vascular Permeability and Systemic Blood Pressure

Walter N. Duran

Department of Pharmacology & Physiology; New Jersey Medical School; Rutgers, The State

University of New Jersey; Newark, NJ 07013, U.S.A.

Nitric oxide (NO) derived from endothelial nitric oxide synthase (eNOS) plays a significant role

in vascular health and disease. Our work focuses mainly on NO as a fundamental signal in

developing a robust increase in endothelial permeability (hyperpermeability) to macromolecules.

We demonstrated that mice genetically lacking eNOS (eNOS-/-) did not develop

hyperpermeability in response to platelet-activating factor (PAF), a pro-inflammatory

phospholipid. Acetylcholine failed to induce vasodilation in eNOS-/- mice. We have also shown

that eNOS undergoes differential translocation from its plasma membrane compartment in

response to specific agonists, and that translocation is a function-targeting mechanism.

Translocation of eNOS to the cytosol serves to initiate hyperpermeability, while translocation to

Golgi is associated with vasodilation. We demonstrated that the onset of hyperpermeability is

associated in part to signaling through the activation of soluble guanylyl cyclase (with

production of cGMP) and in part to NO-induced S-nitrosation of junctional proteins - such as

p-120 catenin and beta-catenin. In addition to permeability studies, evidence will be presented

in support of the hypothesis that acupuncture and the Chinese herb Danshen reduce

hypertension by increasing the expression and the activity of eNOS. (Supported by NIH grants

5RO1 HL070634 and 5RO1 HL088479).


Dr. Walter N. Duran received his Ph.D. degree from

Duke University in 1974. He began his US academic

career as Assistant Medical Research Professor at Duke

University Medical School. Dr. Duran joined New

Jersey Medical School in July 1977 as Assistant

Professor in Physiology and was promoted to Associate

Professor with tenure in July 1979 and to Professor in

1988. Since 1990, he holds a joint appointment as

Professor in Surgery. Since May 2008, he serves as Vice Chair in the Department of

Pharmacology and Physiology. On July 1, 2013, New Jersey Medical School became a part of

Rutgers, The State University of New Jersey.

Dr. Duran’s research focuses on signaling mechanisms that regulate microvascular transport,

with emphasis on endothelial cells and the functional significance of the signaling cascades

associated with endothelial nitric oxide synthase. Dr. Duran’s research efforts have been

supported by NIH since 1981. His research career was assisted also by grants-in-aid from the

American Heart Association, and through an AHA-Established Investigatorship Award.

He served as member of AHA-Peer Review Committees and of the AHA-National Research

Program Evaluation Committee, where he chaired the Peer Review Subcommittee. Dr. Duran

is a member of the American Physiological Society (APS) and has served as a member of

several APS committees. He is also a member of The Microcirculatory Society (MCS), where

he served as member of several committees and as President (2000-2001).

Dr. Duran has also served as a regular member of four NIH Study Sections; he chaired the

NIH-CSR Hypertension and Microcirculation Study Section (2004-2006), and currently serves

on the Surgery, Anesthesiology and Trauma Study Section. He is currently also a member of

the editorial board of specialty journals: Microvascular Research, American Journal of

Physiology: Heart & Circulatory Physiology, and Circulation Research.

Dr. Duran’s research efforts have been recognized through the 1978 MCS Pharmacia Travel

Award, the 1995 UMDNJ University Excellence Award in Biomedical Research, a 2003

Fulbright Research/Teaching Scholarship, the 2003 MCS Eugene M. Landis Award for

Excellence in Research, the 2009 Foundation of the UMDNJ Excellence in Research Award,

and the 2011 Asian Congress for Microcirculation Award for Excellence and Service (Bangkok,

Thailand).

Plenary Lecture 8



Mitochondria as Sensor and Generator of Signals for the Control of

Metabolic Homeostasis

Pierre Maechler, Ph.D.

Professor, Department of Cell Physiology and Metabolism,


Chair: Jing-Yan Han, M.D., Ph.D.


Western Medicine/Tasly Microcirculation Research Center, Peking

University Health Science Center

PL-8

Mitochondria as Sensor and Generator of Signals for the Control of

Metabolic Homeostasis

Pierre Maechler

Department of Cell Physiology and Metabolism, University of Geneva Medical Centre, rue

Michel-Servet 1, 1211 Geneva 4, Switzerland, Email: [email protected]

In the endocrine fraction of the pancreas, the task of the beta-cell is to continuously and

perfectly adjust insulin secretion to fluctuating blood glucose levels, thereby maintaining

glycemia and nutrient homeostasis. This glucose sensing coupled to insulin exocytosis depends

on transduction of metabolic signals into intracellular messengers recognized by the exocytotic

machinery. Central to this metabolism-secretion coupling, mitochondrial signal transduction

refers to both integration and generation of metabolic signals, connecting glucose sensing to

insulin exocytosis. In response to a glucose rise, nucleotides and metabolites are generated by

mitochondria and participate, together with cytosolic calcium, in the stimulation of insulin

release. The presentation will describe the role of mitochondria in metabolic signal transduction

regulating insulin secretion with a special focus on the role of glutamate and its closely

associated enzyme glutamate dehydrogenase (GDH). GDH is a mitochondrial enzyme playing a

pivotal role between carbohydrate and protein metabolisms, controlling production and

consumption of the messenger molecule glutamate in neuroendocrine cells. GDH activity is

under the control of several regulators, conferring to this enzyme energy-sensor property. In

pancreatic beta-cells, inhibition of GDH activity decreases insulin release, while activating

mutations are associated with a hyperinsulinism syndrome. Although GDH enzyme catalyzes

the same reaction in every tissue, its function regarding metabolic homeostasis varies greatly

according to specific organs. Newly generated mouse models lacking GDH in specific tissues

offer the opportunity to decipher organ specificities of GDH regulation and glutamate

pathways.

Prof. Pierre Maechler (Ph.D.) Address: Department of Cell Physiology and Metabolism,

Faculty of Medicine, University of Geneva, 1 rue Michel-Servet, CH-1211 Geneva 4, Switzerland.

Phone + 41 22 379 55 54, Email: [email protected], Web site: http://www.maechler.unige.ch/ Education/diploma:1982-1986: Bachelor, University of Geneva, Faculty of Sciences, section of Biology. 1985-1986: Master degree: laboratory of Prof. M. Schapira under the supervision of Prof. U.K.

Laemmli: “Activation of human neutrophils by plasma kallikrein.” 1990-1993: Ph.D. thesis at Symphar-ILEX Pharmaceutical Research Laboratory (Versoix/San

Antonio) delivered by the University of Geneva, “Study on the mechanisms leading to diabetes-associated hypercholesterolemia in rats with experimental diabetes”.

2002: Privat Docent thesis (lecture on September 23rd): “Mitochondria as the conductor of metabolic signals for insulin exocytosis in pancreatic β-cells.”

Professional experience:1987-1995: Research Scientist at Symphar-ILEX Pharmaceutical Research Laboratory,

Versoix/San Antonio. 1995-2000: Senior Scientist (Maître-Assistant) at the Division of Clinical Biochemistry,

University Medical Center, Geneva. 2001-2006: Head of a junior group, fellow of the Dr Max Cloetta foundation: Maître

d'Enseignement et de Recherche Suppléant (SMER) at the Dept. of Cell Physiology and Metabolism, University Medical Center, Geneva.

2006-2011: Associate Professor at the Dept. of Cell Physiology and Metabolism, University Medical Center, Geneva.

2011-present: Full Professor at the Dept. of Cell Physiology and Metabolism, University Medical Center, Geneva.

Bibliometric data Peer-reviewed articles: 92 h-index: 34 Times cited: 3675

Awards 2000: Paul Langerhans Prize from the European Association for the Study of Diabetes 2001: Recipient of a career development award by the Cloëtta foundation 2001: Recipient of the 2001 research prize from the Swiss Diabetes Foundation. The 2002 Leenaards prize (Lausanne) The 2003 Bizot Price awarded by the Faculty of Medicine (Geneva). The 2003 prize of the Fondation Endocrinologie (Geneva). The 2005 prize of the Fondation Romande pour la Recherche sur le Diabète.

mailto:[email protected]

http://www.maechler.unige.ch/

Plenary Lecture 9



β-Adrenergic Receptor Subtype Signaling in the Heart: From

Bench to the Bedside

Anthony Yiu Ho Woo, Ph.D.

Visiting Associate Professor, Institute of Molecular Medicine,

Peking University

Chair: Hong-Quan Zhang


P

L-9

β-Adrenergic Receptor Subtype Signaling in the Heart: From Bench to

the Bedside

Anthony Yiu Ho Woo, Ph.D. and Rui-Ping Xiao, M.D., Ph.D.

Institute of Molecular Medicine, Peking University, Beijing 100871, China

　β-adrenergic receptor (βAR) stimulation by the sympathetic nervous system or

circulating catecholamines is broadly involved in peripheral blood circulation, metabolic

regulation, muscle contraction, and central neural activities. In the heart, acute βAR stimulation

serves as the most powerful means to regulate cardiac output in response to a fight-or-flight

situation, whereas chronic βAR stimulation plays an important role in physiological and

pathological cardiac remodeling.

There are three βAR subtypes, β1AR, β2AR and β3AR, present in cardiac myocytes. Over

the past two decades, we have systematically investigated the molecular and cellular

mechanisms underlying the distinctly different even opposite functional roles of β1AR and

β2ARsubtypes in regulating cardiac structure and function, with keen interest in the

development of novel therapies based on our bench discoveries. We have made three major

discoveries, including (a) dual coupling of β2AR to Gs and Gi proteins in cardiomyocytes, (2)

cardioprotection by β2AR signaling in improving cardiac function and myocyte viability, and (3)

PKA-independent, CaMKII-mediated βAR apoptotic and maladaptive remodeling signaling in

the heart. Based on our bench discoveries and salutary effects of βAR blockade on patients

with heart failure, we envision that activation of β2AR in combination with clinically used

β1AR blockade should provide a safer and more effective therapy for the treatment of heart

failure.

Anthony Yiu-Ho Woo, Ph.D.

Anthony Yiu-Ho Woo, Ph.D. was graduated in 1993 from the Department of Biochemistry at

The Chinese University of Hong Kong. After obtaining his Master degree in Biochemistry in

1998, he began his career as a research assistant in a project for the “Development of

over-the-counter Chinese herbal pharmaceutical products” in the School of Chinese Medicine in

The Chinese University of Hong Kong. He obtained a Diploma in Pharmaceutical

Management in Chinese Medicine (School of Professional and Continuing Education, The

University of Hong Kong) in 2002. He pursued on his Ph.D. study from 2000 to 2004 at The

Chinese University of Hong Kong focusing on the study of the cardioprotective effects of

Chinese medicinal materials. He received his post-doctoral training in Dr. Rui-Ping Xiao’s

laboratory in National Institute on Aging, National Institute of Health in the United States from

2005 to 2010. He is presently a Visiting Associate Professor in the Laboratory of Signal

Transduction, The Institute of Molecular Medicine at Peking University in Beijing, China.

Dr. Woo’s research interests include the signal transduction of beta adrenergic receptors in the

cardiovascular system and drug development.

Symposium 1

14:05-16:10 September 6 Friday Room 414 Yifu Building

Permeability

Chairs: Jerome W. Breslin

Department of Molecular Pharmacology and Physiology Morsani College of Medicine, University of South Florida

Masato Yasui Department of Pharmacology, School of Medicine, Keio University

S1-1 Thrombin and sphingosine-1-phosphate alter local lamellipodia dynamics to modulate

endothelial barrier function Jerome W. Breslin Department of Molecular Pharmacology and Physiology University of South Florida, Morsani College of Medicine

S1-2 Nitric oxide/Caveolin-1/MMP pathway: a novel therapeutic strategy for drug discovery from herbal medicine targeting blood-brain-barrier disruption during cerebral ischemia-reperfusion injury Jian-Gang Shen School of Chinese Medicine, University of Hong Kong

S1-3 The effects of sphingosine 1-phosphate on modulation of endothelial barrier function Qiao-Bing Huang Department of Pathophysiology, Key Lab for Shock and Microcirculation Research, Southern Medical University

S1-4 Salvianolic acid B binds to Src and ameliorates hyperpermeability of mesenteric venules in rats with endotoxmia Chun-Shui Pan Tasly Microcirculation Research Center, Peking University Health Science Center

S1-5 Analysis of aquaporin-mediated diffusional water permeability by coherent anti-stokes raman scattering microscopy Masato Yasui Department of Pharmacology, Keio School of Medicine

S1-6 CARS microscope: a novel approach to observe epithelial water movement Ying-Chun Yu Department of Pharmacology, Keio University School of Medicine

S1-1 Thrombin and Sphingosine-1-Phosphate Alter Local Lamellipodia Dynamics to Modulate Endothelial Barrier Function Jerome W. Breslin Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida Thrombin and sphingosine-1-phosphate (S1P) increase and decrease endothelial permeability, respectively, by modulating activities of Rho-family GTPases that control the actin cytoskeleton and integrity of intercellular junctions. Our recent finding that endothelial cells in tightly confluent monolayers frequently display local lamellipodia led us to test the hypothesis that these dynamic adhesive structures are important for control of endothelial barrier function. The endothelial barrier was assessed either by measuring transendothelial electrical resistance (TER) of confluent human umbilical vein endothelial cells (HUVEC), or by determining the apparent permeability to albumin in isolated rat mesenteric venules. We used time-lapse microscopy to monitor movement of structures containing GFP-actin or VE-cadherin-GFP expressed in HUVEC. The active, GTP-bound forms of RhoA, Rac1, and Cdc42 were detected by ELISA. The role of Rac1 was also tested, using the selective inhibitor NSC23766, or by transfecting HUVEC with GFP-wild type Rac1 or GFP-dominant negative Rac1. The results show that local lamellipodia persisted for only a few minutes and protruded over adjacent cells, beyond junctions containing VE-cadherin. Thrombin decreased TER and the lamellipodia protrusion frequency, and increased the number of actin filaments after 20-30 min. S1P increased TER and briefly increased the protrusion frequency and turnover time of local lamellipodia. Thrombin increased RhoA-GTP and decreased Rac1-GTP and Cdc42-GTP, while S1P increased both GTP-bound RhoA and Rac1 with no effect on Cdc42. The Rac1 inhibitor NSC23766 decreased lamellipodia formation and HUVEC TER, and also increased permeability of isolated venules. Likewise, HUVEC overexpressing a GFP-dominant negative Rac1 mutant developed a lower TER and had reduced lamellipodia formation compared to those expressing GFP. In contrast, overexpression GFP-wild type Rac1 increased lamellipodia formation and TER. Lastly, specific blockade of lamellipodia formation with the myosin II inhibitor (-)Blebbistatin significantly decreased TER of HUVEC and increased permeability of isolated rat mesenteric venules. Combined, these data suggest that local lamellipodia are 1) influenced by Rac1 activity, 2) require Myosin II activity, 3) are important for the maintenance of normal endothelial barrier integrity, and4) have a role in the thrombin and S1P-induced changes in endothelial permeability.

S1-2 Nitric Oxide/Caveolin-1/MMP Pathway: A Novel Therapeutic Strategy for Drug Discovery from Herbal Medicine Targeting Blood-brain-barrier disruption during Cerebral Ischemia-reperfusion Injury Jian-Gang Shen1,2, Yong Gu1, Ke-Jian Liu2

1. School of Chinese Medicine, University of Hong Kong, Hong Kong, China; 2. College of Pharmacy, University of New Mexico, Albuquerque, New Mexico, USA Aims: Blood-brain-barrier (BBB) disruption is a crucial process in stroke, but no effective drug is available. Thus, drug discovery targeting BBB permeability becomes timely important. In this study, we hypothesized that Nitric oxide (NO)/Cav-1/Matrix metalloproteinase (MMP) pathway is important therapeutic targets in preventing BBB disruption. Following, we investigated the effects of CG, an active compound isolated from Astragali Radix on regulating this pathway and protecting BBB permeability and brain damage during cerebral ischemia-reperfusion injury. Methods: SD rats were subject to different time courses of middle cerebral artery occlusion (MCAO). We investigated the expressions of cav-1 and NOS, the production of NO and peroxynitrite, activation of MMP-9, tight-junction (TJ) protein, BBB permeability and infarction volume in rat and mouse models of cerebral ischemia-reperfusion injury in vivo and hypoxic rat brain microvascular endothelial cells (BMECs) in vitro. CG (60 μmol/kg, i.p.) was used at 15 min before ischemia. Results: (1) NO production reduced cav-1 expression, and the decreased cav-1 was associated with further increases of NO and peroxynitrite production, MMP-2/9 activation, TJ protein degradation and BBB hyper-permeability. (2) L-NAME, a non-selective NOS inhibitor, abolished the cav-1 reduction, MMP-2/9 activations, microvascular hyperpermeability and reduced infarction sizes in the ischemic brains. (3) Cav-1 knockdown by siRNA increased the secretion of MMP-2 to the culture medium. (4) After focal cerebral ischemia-reperfusion, cav-1 deficiency mice displayed higher MMPs activities and BBB permeability than wild-type mice. (5) The effects of L-NAME on MMPs activity and BBB permeability was partly reversed in cav-1 deficiency mice. (6) CG treatment decreased NO and MMPs activation, protected cav-1, reduced infarction volume, BBB permeability and brain damage. Conclusion: Nitric oxide/Cav-1/MMP pathway is an important signal pathway in BBB disruption. The interaction of reactive nitrogen species, cav-1 and MMPs forms a positive feedback loop which provides amplified impacts on BBB dysfunction during cerebral ischemia-reperfusion injury. CG is a potential drug candidate for protecting BBB and reducing infarction volume in ischemic stroke treatment. Acknowledgements: This work is supported by RGC GRF Grant (No. 777611M)

S1-3 The Effects of Sphingosine 1-Phosphate on Modulation of Endothelial Barrier Function Qiang Li, Xiang-Lan Liu, Jing Du, Bo Chen, Li-Qun Wang, Xu-Liang Huang, Qiao-Bing Huang*

Department of Pathophysiology, Key Lab for Shock and Microcirculation Research, Southern Medical University, Guangzhou, 510515, P. R. China Objective：Sphingosine-1-phosphate (S1P)is a bioactive sphingolipid in plasma. S1P is mainly synthesized and secreted by platelets and some other cells. Concentration of S1P can be adjusted through a complex metabolic process, and its change in plasma concentration directly regulates its biological effects. This study was to investigate the effect and the underlying mechanism of different concentration S1P on endothelial barrier function. Methods: Human umbilical vein endothelial cell line (HUVECs) was used in this experiment. Trans endothelial electric resistance (TEER) was detected by resistance meter to clarify the alteration of endothelial barrier function. Distribution of cytoskeleton in HUVECs were observed with immunocytochemistry under confocal microscope. RT-PCR and immunoblotting were used to detect the expression and activation of indicated proteins. Results: Physiological level of S1P could activate S1PR1, then cause endoplasmic reticulum calcium release, resulting in the strengthening of the barrier integrity of endothelial cells by inducing Rac signaling pathway. Appropriate amount of S1P prevented or reversed burn plasma-induced disruption of adherens junction between cultured endothelial cells. The inhibition of S1P synthesis mimicked the burn plasma-evoked redistribution of VE-cadherin and reorganization of F-actin. Both pre- and post-burn applications of certain dose-S1P attenuated the increasing permeability in isolated and perfused skin venules after burn plasma stimulation. Over-dose S1P would bind to S1PR2 and the activation of S1PR2 would cause extracellular calcium influx, leading to destruction of the inter-endothelial junctions by evoking the RhoA and ROCK pathway. The increase expression of S1PR2 played a potential role in endothelial barrier dysfunction under inflammatory situation. Conclusion: S1P is essential in maintaining the basal endothelial barrier function and the imbalance of S1P synthesis and release and the diverse activation of different S1P receptors might affect the vascular permeability under inflammatory situation. [* This work was supported by National Natural Science Foundation of China (No.30971201, No.81170297)]

S1-4 Salvianolic acid B binds to Src and ameliorates hyperpermeability of mesenteric venules in rats with endotoxmia Chun-Shui Pan a,c,d,, Yu-Ying Liu a,c,d, Yu Zhang a,b,c,d, Ke He c,c,d,, Xiao-Yuan Yang a,b,c,d, Bai-He Hu a,c,d, Xin Chang a,c,d, Ming-Xia Wang a,c,d, Xiao-Hong Wei a, Jing-Yu Fan a, and Jing-Yan Han a,b,c,d

a) Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing

100191, China.

b) Department of Integration of Traditional Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing 100191, China.

c) Key Laboratory of Microcirculation, State Administration of Traditional Chinese Medicine of the People's Republic of China, Beijing 100191, China.

d) Key Laboratory of Stasis and Phlegm, State Administration of Traditional Chinese Medicine of the People's Republic of China, Beijing 100191, China.

Background: Microvascular hyperpermeability is a crucial contributor to pathogenesis of various diseases, for which the current clinical therapy remains unsatisfied. Src regulates the hyperpermeability-related proteins, including caveolin-1, VE-cadherin and Zonula occluden-1 (ZO-1). This study aimed to evaluate whether salvianolic acid B (SalB) binds to Src to regulate caveolin-1, VE-cadherin and ZO-1, and ameliorates hyperpermeability of mesenteric venules in rats with endotoxmia. Methods: The male Wistar rats were challenged by infusion of LPS (2 mg/kg/h) for 90 min, with or without SalB (5 mg/kg/h). Human umbilical vein endothelial cells were incubated with LPS or/and SalB. Microcirculation was assessed by intravital microscopy, caveolae in microvascular endothelial cells by electron microscopy, endothelial cell junctional proteins, caveolin-1 and Src by Western blot and confocal microscopy, and the interaction of Src and SalB by Surface Plasmon Resonance (SPR) and Bio Layer Interferometry (BLI). Results: SalB was able to bind to Src in a dose-dependent manner and inhibit the phosphorylation of Src, caveolin-1 and VE-cadherin and restore the distribution of ZO-1 and degradation of VE-cadherin in human umbilical vein endothelial cells exposed to LPS. Furthermore, SalB alleviated the increase of caveolae in endothelial cells and albumin leakage from venules in rat mesentery with endotoxmia. Conclusions: SalB prevents endothelial injury and hyperpermeability via binding to Src inhibiting the activity of Src. These findings suggest SalB to be a promising approach to protect endothelial barrier, and indicate Src as a novel target for vascular hyperpermeability treatment.

S1-5 Analysis of Aquaporin-Mediated Diffusional Water Permeability by Coherent Anti-Stokes Raman Scattering Microscopy Masato Yasui Department of Pharmacology, Keio School of Medicine

Water can pass through biological membranes via two pathways: simple diffusion through the

lipid bilayer, or water-selective facilitated diffusion though aquaporins (AQPs). Although

AQPs play an important role in osmotic water permeability (Pf), the role of AQPs in diffusional

water permeability remains unclear because of the difficulty in measuring diffusional water

permeability (Pd). Here, we report an accurate and instantaneous method for measuring the Pd

of a single mammalian cell using coherent anti-Stokes Raman scattering (CARS) microscopy

with a quick perfusion device for H2O/D2O exchange. The time constant, CARS, for the external

solution H2O/D2O exchange was 16.1 ms, whereas the intracellular H2O/D2O exchange was

100.7 ± 19.6 ms. To evaluate the roles of AQP in diffusional water permeability, AQP4 fused

with enhanced green fluorescent protein (AQP4-EGFP) were transiently expressed in HeLa S3

cells. The time constant, CARS, for the intracellular H2O/D2O exchange in the AQP4-EGFP-

HeLa S3 cells was 43.1 ± 15.8 ms. We also assessed the cell volume and the cell surface area to

calculate Pd. The average Pd values for the AQP4-EGFP- HeLa S3 cells and the control EGFP-

HeLa S3 cells were 2.7 ± 1.0 x 10-3 and 8.3 ± 2.6 x 10-4 cm/s, respectively. AQP4-mediated

water diffusion was independent of the temperature but was dependent on the expression level

of the protein at the plasma membrane. These results suggest the possibility of using CARS

imaging to investigate the hydrodynamics of single mammalian cells as well as the regulation

of AQPs.

S1-6 CARS Microscope: A Novel Approach to Observe Epithelial Water Movement Ying-Chun Yu1, Yoshiro Sohma1, Shinichi Takimoto2, Takayuki Miyauchi1, and Masato

Yasui1*

1 Department of Pharmacology, Keio University School of Medicine, Tokyo, Japan 2 Olympus Corporation, Tokyo, Japan

Coherent Anti-Stokes Raman Scattering (CARS) laser-scanning microscopy gives us an image

of the signal originated from vibration energy of inter-atomic bonds. Studying diffusive water

movements without any staining is useful in developing the mechanism of water transport in

epithelial biology. Observing the O-H stretch vibration with a rapid switching of the perfusion

system from H2O-based isotonic solution to D2O-based one and vice versa, CARS microscope

provided H2O images of biological cells and tissues, whereas no signal was released from D2O

under the same condition. It suggests that it could detect the water movement across cell

membranes or epithelia without any staining.

CARS data provided a time-lapse image of the H2O/D2O exchange and we analyzed the data

with a simulation model consisting of multiple shell-shape compartments. This method

succeeded to determine the diffusional water permeability (Pd) of luminal, basal, and

paracellular pathways in the three dimensional (3D) cyst. 3D-cysts were formed by MDCK

cells in the matrigel and succeeded to make a direct observation of H2O/D2O exchange process

across the MDCK cyst epithelia. Thus the combination of the CARS imaging and the computer

simulation is expected to enable a quantitative description for the water movement across

transport epithelia.

We will report our recent progress in the development of this novel method for measuring the

water diffusional movement.

Symposium 2

8:30-9:50 September 7 Saturday Room 414 Yifu Building

Metabolic Cardiovascular injury and microcirculation

Chairs:

Pierre Maechler

Department of Cell Physiology and Metabolism, University of Geneva

Fu-Long Liao

Institute of Chinese Materia Medica, China Academy of Traditional Chinese Medicine

S2-1 A novel compound CX08005, targeting on PTP1B, improves hepatic microcirculation

disturbance, insulin resistance and hypercholesteremia in pre-diabetic mice

Fei Ye

Beijing Key Laboratory of New Drug Mechanism of Action and Pharmacodynamic

Evaluation, Chinese Academy of Medical Sciences, Beijing Union Medical College,

Institute of Meteria Medica

S2-2 Notoginsenoside R1 protects heart from ischemia reperfusion injury via energy

regulation mechanism

Li Yan


S2-3 A 1H NMR-based metabonomic investigation of time-related metabolic trajectories of

the plasma, urine and liver extracts of hyperlipidemic hamsters

Hai-Bo Zhu

State Key Laboratory of Bioactive Substance and Function of Natural Medicines &

Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Chinese

Academy of Medical Sciences and Peking Union Medical College

S2-4 Danhong injection accelerates vascular injury repair by restoring microcirculation and

promoting endothelial progenitor cell-mediated angiogenesis.

Yan Zhu

Tianjin State Key Laboratory of Modern Chinese Medicine

Tianjin University of Traditional Chinese Medicine

S2-1 A novel compound CX08005, targeting on PTP1B, improves hepatic microcirculation disturbance, insulin resistance and hypercholesteremia in pre-diabetic mice Xiao-Ling Zhang, Juan Li, Li-Wei Huang, Yuan-Feng Tong, Song Wu, Fei Ye Beijing Key Laboratory of New Drug Mechanism of Action and Pharmacodynamic Evaluation, Chinese Academy of Medical Sciences, Beijing Union Medical College, Institute of Meteria Medica, Beijing 10050, China Background: Progression of the metabolic syndrome usually begins with obesity and/or insulin resistance; then, the metabolic risk factors are often marginally increased, like hyperlipidemia, impaired glucose tolerance, diabetes and its complications. Microcirculation disturbance is one of the main pathophysiological processes of metabolic syndrome. It is recognized that disturbances in microvascular functionality are present in early stage. Protein tyrosine phosphatase 1B (PTP1B) is the main negative regulator of insulin signaling. Hence this enzyme represents the potential target for anti-diabetes drugs. However, the effect of PTP1B inhibitor on microcirculation disturbance is unclearly. Objective: To explore the effects of a novel compound CX08005, which inhibits PTP1B activity in vitro and in vivo, on hepatic microcirculation, insulin resistance and hypercholesteremia in mice. Methods: The purity of the compound CX08005 is more than 97%. Recombinant human GST-PTP1B protein was overexpressed by hGST-PTP1B-BL21 E. Coli and purified by GST affinity chromatography. The enzymatic activity was measured by the pNPP assay. Dialysis assay was performed to verify the combine mode between compound CX08005 and PTP1B protein. And the expression of PTP1B in liver of mice was estimated by Western blotting. The pre-diabetic mice were induced by high fat diet in C57BL/6 mice. The compound CX08005 (100 mg/kg body weight/day) was administered orally. The insulin sensitivity was determined by insulin tolerance test (ITT), glucose tolerance test (GTT), and the value of glucose infuse rate (GIR) in hyperinsulinemic-euglycemic clamp test. The insulin response in liver was evaluated by a fluorescence 2-NBDG traced hyperinsulinemic-euglycemic clamp technique. Hepatic microcirculation was observed by inverted microscopy, and the red blood cell (RBC) velocity, sinusoids perfusion, rolling leukocytes and adhering leukocytes were estimated. Results: The hGST-PTP1B activity was almost completely blocked by 1×10-5 M CX08005 (IC50=5.98×10-7 M) with an irreversible inhibitory mode in vitro. Otherwise, the PTP1B expression was down-regulated significantly by CX08005 in liver of the pre-diabetic mice. The HFD-induced insulin resistance, combined with impaired glucose tolerance, damaged insulin tolerance, was markedly reversed by the treatment of CX08005 in model mice. In hyperinsulinemic-euglycemic clamp test, the decreased GIR values were significantly enhanced, and the insulin-dependent glucose uptake into the liver was increased significantly in CX08005 treated mice compared with that in model control mice. Furthermore, CX08005 could significantly increase the blood flow velocity in central and interlobular veins by 86.6% and 104.3%, increase the shear rate of the blood flow by 77.8% and 136.2%, respectively. The number of perfused sinusoids was increased by 49.4%, and the adhered leukocytes both in the center veins and in the hepatic sinusoids area were decreased. Additionally, the hypercholesteremia in model mice was reduced by 20.2% after the CX08005 administration. Conclusion: Compound CX08005, targeting on PTP1b, could be a potential anti-diabetes drug through not only enhancing the insulin sensitivity but also improving the hepatic microcirculation disturbance.

S2-2

Notoginsenoside R1 protects heart from ischemia reperfusion injury via energy regulation mechanism Li Yan1,3, Chun-Shui Pan1,3,Yu-Ying Liu1,3 and Jing-Yan Han1,2,3

1. Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China 2. Department of Integration of Chinese and Western Medicine, School of Basic Medicine Sciences, Peking University, Beijing, China 3. Key Laboratory of Microcirculation, State Administration of Traditional Chinese Medicine,China

Objective: This study was designed to investigate the protective effect of NG-R1 against I/R induced myocardial injury and microvascular hyperpermeability, with focusing on possible involvement of energy metabolism modulation in its action.

Methods: SD rats were subjected to 30 min occlusion of the left anterior descending coronary, followed by reperfusion. NG-R1 was intravenous injected starting from 30 min before ischemia to the end of experiment at the speed of 5 mg/kg·h. MBF, myocardial infarct size, cardiac function, RBC velocity, albumin leakage, ICAM-1 and CD18 were evaluated. Myocardial structure and myocardial apoptosis were assayed by immunofluorescence staining of F-actin and TUNEL. Contents of ATP, ADP, AMP, MPO and MDA in myocardium were determined by ELISA. Expression of ATP 5D, p-Src, and Caveolin-1, ZO-1, Claudin-5 and VE-cadherin were determined by western blot.

Results: Pretreatment with NG-R1 ameliorated I/R-induced MBF decrease, cardiac dysfunction, myocardial RBC velocity reduction, albumin leakage increase. Myocardial infarction size, the percentage of TUNEL positive cells, MPO, MDA, ICAM-1 and CD18 level, increased significantly after I/R, and it was ameliorated by NG-R1. I/R caused decrease in the ratio of ATP/ADP and ATP/AMP, accompanying with reduction of ATP 5D protein and mRNA, which implicated in energy metabolism were significantly alleviated by pretreatment with NG-R1. Moreover, NG-R1 ameliorated I/R-induced P-Src and Caveolin-1 level increase, ZO-1, Claudin-5 and VE-cadherin level decrease.

Conclusions: NG-R1 prevents I/R induced cardiac malfunction and microvascular hyperpermeability, maintains the integrity of myocardial structure through regulating energy metabolism.

S2-3 A 1H NMR-Based Metabonomic Investigation of Time-Related Metabolic Trajectories of the Plasma, Urine and Liver Extracts of Hyperlipidemic Hamsters

Chun-Ying Jiang, Kang-Min Yang, Liu Yang, Zhao-Xia Miao, Ying-Hong Wang*, Hai-Bo Zhu*

State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China The hamster has been previously found to be a suitable model to study the changes associated with diet-induced hyperlipidemia in humans. Traditionally, studies of hyperlipidemia utilize serum- or plasma-based biochemical assays and histopathological evaluation. However, unbiased metabonomic technologies have the potential to identify novel biomarkers of disease. Thus, to obtain a better understanding of the progression of hyperlipidemia and discover potential biomarkers, we have used a proton nuclear magnetic resonance spectroscopy (1H-NMR)-based metabonomics approach to study the metabolic changes occurring in the plasma, urine and liver extracts of hamsters fed a high-fat/high-cholesterol diet. Samples were collected at different time points during the progression of hyperlipidemia, and individual proton NMR spectra were visually and statistically assessed using two multivariate analyses (MVA): principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). Using the commercial software package Chenomx NMR suite, 40 endogenous metabolites in the plasma, 80 in the urine and 60 in the water-soluble fraction of liver extracts were quantified. NMR analysis of all samples showed a time-dependent transition from a physiological to a pathophysiological state during the progression of hyperlipidemia. Analysis of the identified biomarkers of hyperlipidemia suggests that significant perturbations of lipid and amino acid metabolism, as well as inflammation, oxidative stress and changes in gut microbiota metabolites, occurred following cholesterol overloading. The results of this study substantially broaden the metabonomic coverage of hyperlipidemia, enhance our understanding of the mechanism of hyperlipidemia and demonstrate the effectiveness of the NMR-based metabonomics approach to study a complex disease. Key Words: 1H NMR; Metabonomia; Plasma; Urine; Liver; Hamsters * Correspondence Authors: Ying-hong Wang*, Hai-bo Zhu* E-mail: [email protected] (YW); [email protected] (HZ) These authors contributed equally to this work.

S2-4 Danhong Injection Accelerates Vascular Injury Repair by Restoring Microcirculation and Promoting Endothelial Progenitor Cell-mediated Angiogenesis Yan Zhu Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine Center for Research and Development of Chinese Medicine, Tianjin International Joint Academy of Biotechnology & Medicine Ischemic cardiovascular and cerebrovascular diseases are a series of clinical syndromes whose etiology is closely associated with damage of cardiovascular or cerebrovascular walls. Drugs with the effect of promoting blood circulation and removing blood stasis are used to treat the ischemic cardiovascular and cerebrovascular diseases. Danhong injection (DHI) is a preparation composed of Chinese herbs red sage root (Savia miltiorrhiza) and safflower (Carthamus tinctorius L.). Endothelial cells (ECs) and Endothelial Progenitor Cells (EPCs) play an important role in the prevention and treatment of cardiovascular and cerebrovascular diseases, as they promote endothelial growth and neovascularization in vascular repair of ischemic cardiovascular and cerebrovascular diseases. However, the molecular mechanism of how DHI treat the ischemic cardiovascular and cerebrovascular diseases is still not well understood. To address this question, this study exerts a series of experiments to evaluate the repair effect and mechanisms of DHI on vascular injury in connection with the EPCs. In this study, rat hindlimb ischemia model were established to investigate the effect and mechanisms of DHI on microcirculation. Therapeutic doses of Danhong injection were indeed capable of accelerating the recovery of hindlimb blood flow in the ischemic rat model. This action may result from the combined effect of DHI in increasing the capillary density in the hindlimb gastrocnemius of the ischemic rat model and improving expression of chemokine receptor CXCR4. The effect of DHI on proliferation, migration, adhesion, and angiogenesis in ECs and EPCs were also studied. DHI promoted the EPCs proliferation, and improve some functions of migration, adhesion, and tube formation in vitro in the EA.hy 926 and EPCs. Finally, “angiogenic gene array” using QPCR revealed that appropriate dose of DHI upregulated gene expression of several key regulators, including 3 growth factors (VEGF A, VEGF C and endothelin 1)，three receptors and adhesion molecules (CXCR 4, KDR and Integrin αv)，and two enzymes related to angiogenesis (eNOS and MMP 9). At the same time, we found that some factors such as ANG, such as ANG, FGF 2, PECAM 1, MMP 2, MMP 14, COX-1 and AKT-1 could not be upregulated. In conclusion, our results showed that DHI is capable of acting on mature and progenitor Ecs, promoting the functions related to angiogenesis such as cell proliferation, migration, adhesion and cell tube formation in vitro. Based on these findings, DHI can accelerate the recovery of hindlimb blood flow by increasing the capillary density in the hindlimb gastrocnemius of the ischemic rat model. The mechanisms of DHI may at least be mediated by the activation of the SDF-1α/CXCR4, VEGF/KDR and eNOS/MMP 9 signal pathways.

Symposium 3


Organ injury and microcirculation

Chairs:

Mack Wu

Department of Molecular Pharmacology and Physiology, University of South Florida

Morsani College of Medicine

Qiao-Bing Huang

Department of Pathophysiology, Key Lab for Shock and Microcirculation Research,

Southern Medical University

S3-1 Pharmacological studies of mudanpi and its main active component on cardiac ischemia

and ion channels

Yu-Ling Ma

Department of Physiology, Anatomy and Genetics, University of Oxford

S3-2 Tissue optical clearing for high resolution blood flow monitoring with laser speckle

contrast imaging

Dan Zhu

Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for

Optoelectronics

S3-3 3, 4-dihydroxyl-phenyl lactic acid restores NADH dehydrogenase [ubiquinone] 1 alpha

subcomplex subunit 10 expression ameliorating cardiac reperfusion injury

Xiao-Yuan Yang


S3-4 Lymphatic drainage pathway from CSF is impaired after subarachnoid hemorrhage

Bao-Liang Sun

Key Lab of Cerebral Microcirculation in Universities of Shandong (Taishan Medical

University); Department of Neurology, Affiliated Hospital of Taishan Medical

University

S3-1 Pharmacological studies of Mudanpi and its main active component on cardiac ischemia and ion channels Yu-Ling Ma, Penelope J. Noble, Soojin Lee, Robert Wilkins, Clive Ellory and Denis Noble Department of Physiology, Anatomy and Genetics, University of Oxford, Parks Road, Oxford OX1 3PT Mudanpi, a herbal medicine prepared from the root bark of Paeonia Suffruticosa, was traditionally used for thousands of years to ‘remove pathogenic heat from blood’. Previous studies showed that Mudanpi has a significant cardioprotective effect against cardiac ischemia (Ma et al 1984), and its active component, paeonol, inhibits the pacemaker potential of cultured myocytes (Ma et al 1986) suggesting that paeonol might influence the activity of cardiac ion channels. Further studies indeed showed that paeonol has a significant effect on the ion channels of cardiac myocytes (Ma et al 2006). We present here the findings obtained over many years from the East to the West; data were collected from heart ischemic model to ion channel activities of the isolated cardiac myocytes using patch clamp electrophysiological method. The cardiac ischemic model was established by coronary artery occlusion in canines; extract of mudanpi was infused intravenously. The epicardial ECG were recorded by applying 30 electrodes to predetermined epicardial sites. The electrodes were connected to an amplifier, and the epicardial ECG recorded. The elevation of S-T segments were used as a measure of the degree of ischemia. Aterial pressure, heart rate and heart oxygen consumption were recorded. The actions of paeonol on ionic channel were studied using the standard whole-cell configuration of the patch-clamp technique in isolated guinea-pig ventricular myocytes. The effects of paeonol on the action potential (AP) and a variety of ion channels were studied, and the possible mechanisms underlying the alteration of ion channel function were analysed. The results showed that Mudanpi had a significant protective effect against acute ischemic injury by reducing myocardial oxgen comsumption, reducing cardiac arrhythmic risk, lowering blood pressure and heart rate. The main effect of paeonol on the ventricular myocytes was to shorten the AP duration, suppress the AP upstroke velocity and amplitude, actions associated with the blockade of the voltage-gated, fast sodium current. The shortening effect of paeonol on the AP duration was independent of the blockade of the fast sodium current, and the blockage of calcium current, nor did it activate ATP-sensitive potassium current or chloride current. However, paeonol did block the slowly inactivating sodium current, and blockade of this current is known to result in AP shortening. Using the Noble Heart computational model (Noble et al 1998) we predicted that paeonol inhibits ADP of the AP, a strong indication of antiarrythmic action. These findings suggest that paeonol, and therefore Mudanpi, may possess antiarrhythmic activity, which may confer their cardioprotective effects. Acknowledgement We thank Professor Lian-da Li and Professor Alison Gurney for supervising Yu-Ling’s MSc and PhD studies in making the findings presented here. References Ma, YL., Bates, S. and Gurney, A. (2006). European Journal of Pharmacology. 545:87-92 Noble, D., Varghese, A., Kohl, P. and Noble, P. (1998). Can J Cardiol, 14: 123-134. Ma Yu-ling, Li Lian-da at al. (1986). Chinese J. Integrated Traditional and Western Medicine. 6(5):292-294 Ma Yu-ling, Li Lian-da et al. (1984). Shan Xi Medical and Pharmaceutical J. 18(4): 212-214

S3-2 Tissue Optical Clearing for High Resolution Blood Flow Monitoring with Laser Speckle Contrast Imaging Dan Zhu Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics Key Laboratory of Biomedical Photonics of Ministry of Education, Department of Biomedical Engineering, Huazhong University of Science and Technology, Wuhan, China E-mail: [email protected] Microvascular structure and function is tightly coupled with health status. For instance, the impairment in the skin microcirculation is one of the main features in advanced clinical stages of the peripheral vascular disease. Understanding of the cerebral blood flow distribution of a given cortex area is of considerably significant for revealing the information processing mechanisms of brain. In order to monitor the dynamics of microvasculature, various imaging techniques have been developed, including Magnetic Resonance Imaging, laser Doppler flowmetry, and Laser Speckle Contrast Imaging (LSCI), etc. The former two lack in temporal-spatial resolution, which cannot provide both structural and functional information of microvasculature simultaneously. In contrast, LSCI, a full-field optical imaging technique, can provide a two-dimensional map of blood flow at high temporal-spatial resolution, which has been playing an important role in studying cerebral blood flow, mesentery microcirculation, and angiogenesis in Chick Chorioallantoic Membrane, but is still limited to transparent tissues based on some window models.. Recently, Tissue Optical Clearing (TOC) technique has shown significant potential for improving biomedical optical imaging. Recently, the combination of in vitro TOC and various microscopical techniques made a breakthrough for the research of neuroscience. In vivo TOC technique needs to meet the follows: fast treating, enough transparent and safe to animals, so its development is relative lagging. Fortunately, some innovative optical clearing methods for in vivo were proposed and enhanced the contrast and resolution of laser speckle contrast imaging (LSCI) for blood flow monitoring. In this presentation, the current status is reviewed, which includes the principle of TOC induced improvement of LSCI, progress in tissue optical clearing for blood flow monitoring, the safety of optical clearing agents to animals. Up till now, various transparent skin windows and cranial window are established by topical application of optical clearing agents instead of surgery, which enable LSCI to monitor dermal or cerebral blood flow with high resolution and contrast. And safety investigations show that it is possible to repeatedly image dermal blood flow by a switchable transparent skin window without side effect. The future development trendy will focus on the safety and widely applications in biomedicine. Key words: blood flow, laser speckle contrast imaging, skin, skull, tissue optical clearing

S3-3 3, 4-dihydroxyl-phenyl lactic acid restores NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 10 expression ameliorating cardiac reperfusion injury Xiao-Yuan Yang1,2, Chun-Shui Pan1, Quan Li1, Yu-Ying Liu1, Li Yan1, Xiao-Hong Wei1, Bai-He Hu1, Xin Chang1, Xiao-Wei Mao1,2, Li-Jun Wang3, Shui-Wang Hu4, Yong Jiang4, Guo-Cheng Wang1, Jing-Yu Fan1, Tai-Ping Fan5 and Jing-Yan Han 1,2,6,7 *

1 Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China. 2 Department of Integration of Traditional Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China. 3 Department of Biophysics, Peking University Health Science Center, Beijing, China. 4 Department of Pathophysiology and Key Laboratory of Proteomics of Guangdong Province, Southern Medical University, Guangzhou, China. 5 Angiogenesis & Chinese Medicine Laboratory, Department of Pharmacology, University of Cambridge, UK. 6 Key Laboratory of Microcirculation, State Administration of Traditional Chinese Medicine of the People's Republic of China. 7 Key Laboratory of Stasis and Phlegm, State Administration of Traditional Chinese Medicine of the People's Republic of China. Traditional Chinese Medicine (TCM) has distinct superiority in treatment of complicated diseases, and identification of the target (s) for TCM component (s) is critically important for disclosing the underlying mechanisms. Our results on rats with cardiac ischemia and reperfusion (I/R) injury demonstrated 3, 4-dihydroxyl-phenyl lactic acid (DLA), an ingredient of TCM cardiotonic pills® that has been scheduled to undergo phase Ⅲ clinical trials for treatment of ischemic cardiovascular disease in U.S, prevented hearts from I/R induced reduction in NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, 10 (NDUFA10) protein and mRNA. DLA blunted I/R induced impairment of Complex Ⅰ activity, reduction in the ratio of ATP/ADP and ATP/AMP, MDA accumulation as well as deleterious alterations of mitochondrial ultrastructure. Furthermore, DLA diminished apoptotic myocardium after I/R by upregulating Bcl-2 and downregulating Bax and cleaved Caspase-3. Administration of DLA reduced infarct size and ameliorated the disordered cardiac function and blood flow by I/R. Moreover, the present study revealed DLA able to bind to SIRT1 and restore SIRT1 expression and activity after I/R. Detection of downstream Foxo-1 acetylation level and MnSOD expression showed that DLA enhanced SIRT1 activity. SIRT1 inhibitor, sirtinol and EX527, blunted the role of DLA in relieving NDUFA10 expression and Complex Ⅰactivity. In addition, DLA derivatives, which were synthesized by replacing both phenolic hydroxyl of DLA into methoxyl or hydroxyl, exerted lower affinity for SIRT1 and impaired ability to decrease I/R caused infarct size compared with DLA. This study clarified the target for DLA, providing insight for better understanding the mechanism behind the role of DLA.

S3-4 Lymphatic drainage pathway from CSF is impaired after subarachnoid hemorrhage Bao-Liang Sun1, Xuan Wang1, Li-li Jia1, Zong-Yong Zhang1, Ming-Feng Yang1, Da-Wei Li1,

Hui Yuan1, Yan-Bo Zhang1, Xiao-Yi Yang1, Jun Chen2

1Key Lab of Cerebral Microcirculation in Universities of Shandong (Taishan Medical

University); Department of Neurology, Affiliated Hospital of Taishan Medical University,

Taian, Shandong 271000, China 2Center of Cerebrovascular Disease research, University of Pittsburgh School of Medicine,

Pittsburgh, Pennsylvania 15261, USA

Objective: To investigate the importance of cerebral lymphatic drainage in the process of

cerebral vasospasm after subarachnoid hemorrhage (SAH).

Methods: In this experiment, lymphatic pathway blockade was made by ligating the cervical

lymphatic input and output tubes and removal of bilateral shallow and deep lymphatic nodes.

Rat SAH models were replicated by double cisterna magna injections of autologus arterial

blood. 100µg 125I labeled human albumin as macromolecular tracer was injected in to rat’s

lateral cerebral ventricle. A radioimmunoassay was used to sequentially detect plasma recovery

of 125I labeled human albumin. Based on the single compartment model, concentration-time

curve was obtained in the experiment. Some pharmacokinetic parameters, including peak

concentration [(Cmax)( µg/ml)], time of maximum concentration [(Tmax)(h)] , and elimination

rate constant [(Ka) (h-1)] were calculated. The clearance of macromolecular tracers in CSF by

lymphtic pathway was evaluated.

Results: It was found that in SAH rat, the lymphatic pathway blockade led to an decrease of

68.82% in Cmax. The lymphatic pathway blockade delayed Tmax by 5.47 hours. Ka was also

decreased by 43.24% after lymphatic pathway blockade. The results suggest that in the

condition of SAH, a good proportion of macromolecular substances in CSF were removed by

lymphatic pathway.

Conclusions: Cerebral lymphatic drainage pathway is impaired after SAH. Improving the

lymphatic drainage pathway of CSF may be a new approach for the therapeutic study on SAH

and related cerebral injury.

Symposium 4


Oxidative stress and microcirculation Chairs:

David Zawieja,

Department of Medical Physiology

Division of Lymphatic Biology


Makoto Suematsu,

Department of Biochemistry and Integrative Medical Biology, School of Medicine,

Keio University

S4-1 ERM protein moesin is phosphorylated by advanced glycation end products and

modulates endothelial permeability

Xiao-Hua Guo

Department of Pathophysiology, Key Lab for Shock and Microcirculation Research,

Southern Medical University

S4-2 Establishment of high-throughput screening for active pharmaceutical ingredients of

acute cerebral ischemia and related applied research

Rui Tan

School of Life Science and Technology, Southwest of Jiaotong University

S4-3 The symphonic mechanism of survival or death of the neurons following ischemia

stroke

Chang-Man Zhou

Department of Anatomy, Peking University Health Sciences Center

S4-4 Curcumin induces autophagy to protect vascular endothelial cell survival from oxidative

stress damage

Xue-Jun Li

Department of Pharmacology, School of Basic Medical Sciences, Peking University

S4-1 ERM protein moesin is phosphorylated by advanced glycation end products and modulates endothelial permeability Xiao-Hua Guo, Ling-Jun Wang, Bo Chen, Qiang Li, Ji-Ping Wang, Ming Zhao, Wei Wu, Xu-Liang Huang and Qiao-Bing Huang Department of Pathophysiology, Key Lab for Shock and Microcirculation Research, Southern Medical University, Guangzhou, People’s Republic of China Advanced glycation end products (AGEs) accumulated in different pathological conditions have the potent capacity to alter cellular properties that include endothelial structural and functional regulations. The disruption of endothelial barrier integrity may contribute to AGE-induced microangiopathy and macrovasculopathy. Previous studies have shown that AGEs induced the rearrangement of actin and subsequent hyperpermeability in endothelial cells (ECs). However, the mechanisms involved in this AGE-evoked EC malfunction are not well understood. This study directly evaluated the involvement of moesin phosphorylation in AGE-induced alterations and the effects of the RhoA and p38 MAPK pathways on this process. Using immortalized human dermal microvascular ECs (HMVECs), we first confirmed that the ezrin/radixin/moesin (ERM) protein moesin is required in AGE-induced F-actin rearrangement and hyperpermeability responses in ECs by knockdown of moesin protein expression with small interfering RNA. We then detected AGE-induced moesin phosphorylation by Western blot analysis. The mechanisms involved in moesin phosphorylation were analyzed by blocking AGE receptor binding and inhibiting Rho and MAPK pathways. AGE-treated HMVECs exhibited time- and dose-dependent increases in the Thr 558 hosphorylation of moesin. The increased moesin phosphorylation was attenuated by preadministrations of AGE receptor antibody, Rho kinase (ROCK), or p38 inhibitor. Suppression of p38 activation via the expression of dominant negative mutants with Ad.MKK6b or Ad.p38 also decreased moesin phosphorylation. The activation of the p38 pathway by transfection of HMVECs with an adenoviral construct of dominant active MKK6b resulted in moesin phosphorylation. These results suggest a critical role of moesin phosphorylation in AGE-induced EC functional and morphological regulations. Activation of the ROCK and p38 pathways is required in moesin phosphorylation. Key words: vascular permeability; receptor for advanced glycation end products; Rho kinase; mitogen-activated protein kinase; ezrin/radixin/moesin

S4-2

Establishment of high-throughput screening for active pharmaceutical

ingredients of acute cerebral ischemia and related applied research

Rui Tan

School of Life Science and Technology, Southwest of Jiaotong University

Ischemic cerebrovascular disease is a disease of the brain blood disorder caused by ischemia,

hypoxia of the corresponding region of the brain tissue, which results in brain damage, bringing

about a series of biochemical metabolic disorders and physiological loss of function. The

pathological response and effective treatment for acute cerebral ischemia mainly involve

adaption of body to hypoxia and ischemia, angiogenesis, neuronal protection and regeneration.

This study intends to conduct work in three aspects: 1 Establish fast, high-throughput screening

cell models for pharmaceutical active ingredients; 2 Use acute cerebral ischemia animal models

to identify drug efficacy in vivo; 3 Establish genetics modified rat brain stem cells for treatment

of acute cerebral ischemia rats. Results that have been obtained: 1 A stable and reliable rat

models of acute cerebral ischemia have been established. 2 Three functional eukaryotic

expression plasmids of HIF1α, VEGF, BDNF have been constructed to relieve core lesion of

cerebral hypoxia, ischemia and neuronal protection / regeneration. 3 HIF1α, VEGF and NGF

gene promoter have been cloned to construct a series of promoter-probe eukaryotic expression

plasmid and will be used to establish Stable cell lines and build a fast, high-throughput

screening system of pharmaceutical active ingredients in the next step. 4 Primary rat bone

marrow mesenchymal stem cells (rMSC) have been isolated and cultured. Next Steps: 1 Use

rMSC and HEK293 cell models of promoter-probe eukaryotic expression plasmids to conduct

drug screening and function confirmation. 2 Use HIF1α, VEGF and BDNF gene-modified stem

cells to treat the acute cerebral ischemia rat models.

S4-3 The orchestrating mechanism of survival or death of the neurons following ischemia stroke

Chang-Man Zhou

Department of Anatomy, Peking University Health Sciences Center, Beijing China 100191

At least three distinct modes of cell death have been identified and implicated with the ischemic

stroke process – they include necrosis, apoptosis, and autophagy. Delayed neuronal death is a

hallmark feature of stroke and the primary target for neuroprotective strategies. This is

evidenced by the fact that, even if cerebral blood flow is re-established quickly enough to

prevent immediate cell death after a stroke, many of the initially surviving neurons still die

hours to days after reperfusion. Autophagy is an essential process for cellular metabolism and

surviva, while on the other hand, it may also be involved in cell death, specifically orchestrating

the effects in cerebral ischemia. Of the many pathophysiological events involved in this delayed

neuronal injury, mitochondria-specific autophagy, also known as mitophagy, has recently been

shown to be a contributing factor. We showed that the HIF-1 and gene BNIP3 caused neuronal

death by inducing mitochondrial degeneration. Our recent preliminary data from BNIP3

knockout mice showed that BNIP3 deficiency robustly reduced ischemia-induced mitophagy in

neurons. BNIP3 appears, therefore, to be a regulator of mitophagy. In characterizing the BNIP3

pathway, we have obtained preliminary data showing that BNIP3 interacts with mitochondria

by binding to the voltage-dependent anion channel.

S4-4 Curcumin induces autophagy to protect vascular endothelial cell survival from oxidative stress damage

Xue-Jun Li

Department of Pharmacology, School of Basic Medical Sciences, Peking University, Beijing,

China

Autophagy is an evolutionarily conserved catabolic mechanism used by most kinds of

mammalian cells that involves the sequestration, transport, bulk degradation and recycling of

cytoplasmic components, such as long-lived proteins and organelles. In mammalian cells,

autophagy acts predominantly as a prosurvival pathway, protecting the cells from oxidative

stress. Our study first proposed that curcumin could protect human endothelial cells from the

damage caused by oxidative stress via autophagy. Furthermore, our results revealed that

curcumin causes some novel cellular mechanisms that promote autophagy as a protective effect.

Pretreatment with curcumin remarkably improves the survival of human umbilical vein

endothelial cells (HUVECs) from H2O2-induced viability loss, which specifically evokes an

autophagic response. Exposed to H2O2, curcumin-treated HUVECs upregulate the level of

LC3-II, the number of autophagosomes, and the degradation of p62. We show that this

compound promotes BECN1 expression and inhibits the phosphatidylinositol 3-kinase

(PtdIns3K)-AKT-mechanistic target of rapamycin (MTOR) signaling pathway. Curcumin can

also reverse FOXO1 (a mediator of autophagy) nuclear localization along with causing an

elevated level of cytoplasmic acetylation of FOXO1 and the interaction of acetylated FOXO1

and ATG7, under the circumstance of oxidative stress. These results suggest that curcumin has

the potential for use as an autophagic-related antioxidant for prevention and treatment of

oxidative stress. These data uncover a brand new protective mechanism involving FOXO1 as

having a critical role in regulating autophagy in HUVECs.

Symposium 5


Inflammation and microcirculation

Chairs: Sarah Yuan

Department of Molecular Pharmacology and Physiology, University of South Florida Morsani College of Medicine

Xue-Jun Li

Department of Pharmacology, School of Basic Medical Sciences, Peking University

S5-1 Sinomenine decreases MyD88 expression and improves inflammation-induced joint

damage progression and symptoms in rat adjuvant-induced arthritis

Hui Cai

Department of integrated traditional and western medicine, School of Medicine, Second

Military Medical University (Shanghai), Jinling Hospital

S5-2 Ginsenoside Rb1 ameliorates caveolae-mediated albumin leakage from rat mesenteric

venules induced by lipopolysaccharide

Kai Sun


S5-3 Andrographolide pills® attenuates lipopolysaccharide-induced pulmonary

microcirculatory disturbance and acute lung injury inr ats

Ning Yang

The Institute of Quality Control of Medical Material and Equipment under the Joint

Logistic Department of Beijing Military Command

S5-4 Role of an integrin-interacting protein in the control of EMT in cancer and fibrosis

Hong-Quan Zhang

Laboratory of Molecular Cell Biology and Tumor Biology, Department of Anatomy,

Histology and Embryology, School of Basic Medical Sciences, Peking University

Health Science Center

S5-1 Sinomenine decreases MyD88 expression and improves inflammation-induced joint damage progression and symptoms in rat adjuvant-induced arthritis

Hui Mu, Ru-Bing Yao, Ling-Jie Zhao, Si-Yu Shen, Zhi-Ming Zhao, Hui Cai*

School of Medicine, Second Military Medical University (Shanghai), Jinling Hospital,

Department of integrated traditional and western medicine, Nanjing, Jiangsu, China

* Corresponding author at: Department of integrated traditional and western medicine, Jinling

Hospital, 305 East Zhongshan Road, Nanjing 210002, Jiangsu Province, China.

E-mail address: [email protected] (Hui Cai).

objective

Sinomenine (SIN) is the active principle of the Chinese medical plant sinomenium acutum

which is widely used for the treatment of rheumatoid arthritis (RA) in China. Recently, several

groups indicated that Myeloid differentiation primary response protein88 (MyD88) might be

associated with disease progression of RA. Here, we mainly studied the effect of SIN on

MyD88 expression and showed its anti-inflammation role in arthritis.

Methods

MyD88 expression and the effect of SIN on its expression were evaluated by determinating the

protein and mRNA levels of MyD88 in the synovial tissues. The anti-inflammation effect of

SIN on AIA were assessed by histological changes and pro-inflammation cytokines expression.

The therapeutic effects of SIN on AIA were evaluated by synovial hypertrophied, cartilage and

joint damage, joint swelling and pain.

results

MyD88 was mainly located in characteristic pathological structures of RA synovial tissues. In

AIA rats, there was an increase in MyD88 expression in the synovial tissues, and SIN markedly

decreased the increase. SIN suppressed synovial inflammation and inflammation-induced joint

destructive progression and arthritis symptoms in AIA rats.

Conclusions

SIN is an effective therapeutic agent for RA. Targeting MyD88 signaling may provide new

methods for the treatment of RA.

Key words: sinomenine; MyD88; adjuvant-induced arthritis; synovial; Rheumatoid arthritis;

inflammation; pain

S5-2 Ginsenoside Rb1 ameliorates caveolae-mediated albumin leakage from rat mesenteric venules induced by lipopolysaccharide

Yu Zhang 1,3,4, Kai Sun 2,3,4, Yu-Ying Liu 2,3,4, Yun-Pei Zhang 1,3,4, Bai-He Hu 2,3,4, Xin-Chang 2,3,4, Li Yan 2,3,4, Chun-Shui Pan 2,3,4, Quan Li 2,3,4, Jing-Yu Fan 2,3,4, Ke He 1,3,4, Xiao-Wei Mao 1,3,4, Lei Tu 1,3,4, Chuan-She Wang 1,2,3,4 and Jing-Yan Han 1,2,3,4*

1 Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China 2 Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China 3 Key Laboratory of Microcirculation, State Administration of Traditional Chinese Medicine of China, Beijing, China 4 Key Laboratory of Stasis and Phlegm of State Administration of Traditional Chinese Medicine, Beijing, China * Reporter

Background: Lipopolysaccharide (LPS) is one of the common pathological factors which causes mesentery hyperpermeability and intestinal edema related diseases. This study evaluated whether ginsenoside Rb1 (Rb1), an ingredient of a Chinese medicine Panax ginseng, has beneficial effects on mesentery microvascular hyperpermeability induced by LPS and the underlying mechanisms.

Methods: Male Wistar rats were continuously infused with LPS (5 mg/kg/hr) via the left jugular vein for 90 min. In some rats, Rb1 (5 mg/kg/hr) was administrated through the left jugular vein 30 min after LPS infusion. The dynamics of fluorescein isothiocynate-labeled albumin leakage from mesentery venules was observed by intravital microscopy. Intestinal tissue edema was evaluated by hematoxylin and eosin staining. The number of caveolae in endothelium of microvessels was examined by electron microscopy. Confocal microscopy and Western blotting were applied to detect caveolin-1 (Cav-1) expression, Cav-1 phosphorylation and concerning signaling proteins in intestinal tissues and human umbilical vein endothelial cells.

Results: LPS infusion evoked an increased albumin leakage from mesentery venules, which was significantly ameliorated by Rb1 post-treatment. Mortality and intestinal edema around microvessels were also reduced by Rb1. Rb1 decreased caveolae number in endothelium of microvessels. Cav-1 expression and phosphorylation, nuclear factor-kappa B (NF-κB) activation and Src kinase phosphorylation were inhibited by Rb1.

Conclusion: Rb1 ameliorated caveolae-mediated albumin leakage after the onset of endotoxemia and improved intestinal edema through inhibiting Cav-1 expression and phosphorylation, which was correlated to suppression of NF-κB and Src activation.

S5-3 Andrographolide pills® Attenuates Lipopolysaccharide-Induced Pulmonary Microcirculatory Disturbance and Acute Lung Injury in Rats Ning Yang 1.2.3,4, Yu-Ying Liu 2, Chun-Shui Pan 2, Kai Sun 2, Xiao-Hong Wei 2, Xiao-Wei Mao 1,2, Fang Lin 2, Xue-Jun Li 3, Jing-Yu Fan 2 and Jing-Yan Han1.2*

1 Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China 2 Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China 3 Department of Pharmacology, School of Basic Medical Sciences, Peking University, Beijing, China 4 The Institute of Quality Control of Medical Material and Equipment under the Joint Logistic Department of Beijing Military Command, Beijing, China Aim: Lipopolysaccharide (LPS)-induced pulmonary micocirculatory disturbance (PMD) is the

pathological basis in LPS-induced ALI, manifesting as leukocytes adhesion, release of peroxide,

and pulmonary microvessel hyperpermeability. Andrographolide pills® (AP) is approved for

treatment of acute upper respiratory tract infection. The purpose of the present study was to explore

the protective effect of AP on LPS-induced PMD and ALI.

Material and Methods: Male Sprague-Dawley rats were continuously infused with LPS (5 mg/kg/

hr) for 1 hour to induce PMD and ALI. AP (0.18 g/kg or 1.8 g/kg) was administrated orally 1 h

before LPS exposure. Arterial blood pressure and heart rate were monitored. Blood gas analysis,

histological observation, cytokines in plasma and PMD included neutrophil adhesion to pulmonary

venules and infiltration into lung tissue, pulmonary oxidative stress, pulmonary microvessel

permeability and edema were evaluated 6 h after LPS challenge. Meanwhile, PMD and ALI related

proteins were assessed simultaneously.

Results: Rats receiving LPS exhibited significant alterations of mean arterial blood pressure and

heart rate, and dramatical increase in mortality and cytokines in the plasma. Neutrophil adhesion

and infiltration in lung tissue, pulmonary oxidative stress and microvessel permeability were all

increased significantly, ultimately leading to pulmonary injury, and lung dysfunction. Pretreatment

with AP (0.18 g/kg or 1.8 g/kg) significantly attenuated all aforementioned insults induced by LPS.

Moreover, Pretreatment with AP significantly inhibited expression of CD11b/CD18 on neutrophil

and intercellular adhesion molecule-1, toll-like receptor 4, caveolin-1, p-caveolin-1, Src, p-Src in

lung tissue induced by LPS, and maintained expression of junctional adhesion molecule-A and

claudin-5.

Conclusion: The results demonstrated that AP showed protective effect on LPS-induced PMD and

ALI, suggesting the potential of AP as a prophylactic strategy for LPS-induced ALI.

S5-4 Role of an integrin-interacting protein in the control of EMT in cancer and fibrosis

Hong-Quan Zhang

Laboratory of Molecular Cell Biology and Tumor Biology, Department of Anatomy, Histology

and Embryology, School of Basic Medical Sciences, Peking University Health Science Center #

38 Xue Yuan Avenue, Beijing 100191, China.

Email: [email protected]

Integrins are transmembrane proteins that link extracellular matrices with cytoskeleton and

mediate a variety of biological functions including cell adhesion and migration as well as tumor

cell invasion and metastasis. In this report, we identified a novel role of an integrin-interacting

protein Kind-2 in the control tumor metastasis in human breast carcinoma via activation of

canonical Wnt/TCF pathway. An array based systematic analyses indicated an overall initiation

of tumor invasion and metastasis program including upregulation of key Wnt/TCF signaling

targets TCF4, LEF1 and HOXB9 as well as epithelial-mesenchymal transition (EMT)

regulatory transcriptional factors Snail, Slug, Twist and Zeb1. The Kind-2/beta-catenin

signaling greatly accelerates tumor cell proliferation, migration, and invasion in vitro, and

promotes tumor growth and lung-metastasis in mice, demonstrating a role of Kind-2 in cancer

progression. On the other hand, we also identified that Kind-2 is involved in the regulation of

TGF-β signaling and promotes renal interstitial fibrosis by inducing EMT. Depletion of Kind-2

in kidney reduces renal interstitial fibrosis in UUO mice including reduced collagen deposition

and decreased α-SMA and fibronectin expressions. Overall, our results indicate an important

role of Kind-2 in the regulation of tumor metastasis and kidney fibrosis via an EMT program.

Symposium 6

8:30-9:50 September 8 Sunday Room 414 Yifu Building

Thrombus

Chairs:

Ke-Sheng Dai



Key Laboratory of Thrombosis and Hemostasis, Ministry of Health Suzhou

Jian-Bo Wu


S6-1 The interaction of GPIb-IX with VWF incurs platelet activation, apoptosis, and GPIbα

ectodomain shedding

Ke-Sheng Dai

Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University

S6-2 Targeting plasminogen activator inhibitor-1 to prevent vascular disease

Jian-Bo Wu


S6-3 Caffeic acid inhibits platelet activation ex vivo and thrombus formation in vivo

Quan Li


S6-4 Trapping red blood cells within microvessels in living mice by optical tweezers

Xun-Bin Wei

School of Biomedical Engineering, Shanghai Jiao Tong University

S6-1 The interaction of GPIb-IX with VWF incurs platelet activation, apoptosis, and GPIbα ectodomain shedding Ke-Sheng Dai



Key Laboratory of Thrombosis and Hemostasis, Ministry of Health

Suzhou 215006, China.

Under high shear rate flow conditions such as in arteries and capillaries, platelet adhesion to the

subendothelium is dependent on interaction between subendothelial-bound von Willebrand

factor (VWF) and its receptor, the platelet glycoprotein Ib-IX-V complex (GPIb-IX-V). It has

been well established that the interaction of GPIb-IX-V with VWF initials platelet adhesion,

activation, and thrombus formation. We found that Ristocetin-induced GPIba–VWF interaction

elicited apoptotic events in platelets, including phosphatidylserine exposure, elevations of Bax

and Bak, gelsolin cleavage, and depolarization of mitochondrial inner transmembrane potential.

Apoptotic events were also elicited in platelets exposed to pathologic shear stresses in the

presence of VWF; however, the shear-induced apoptosis was eliminated by the anti-GPIba

antibody AK2. Furthermore, washed platelets were exposed to VWF-coated glass capillary or

cone-and-plate viscometer at different shear rates, and GPIba ectodomain was shed from

platelets, while a small mass of GPIba COOH-terminal peptide, 17 kDa, was increased

correspondingly. The extent of GPIba shedding was enhanced with the concentration of

immobilized VWF and the time duration of constant shear stress, whereas it was obviously

reduced with the decreased number of adherent platelets. These results indicate the

GPIba–VWF interaction induces apoptotic events in platelets, and results in GPIba ectodomain

shedding. These findings not only have a physiological implication in understanding platelet

clearance and the presence of glycocalicin in normal circulation, but also suggest novel

mechanisms for the negative regulation of platelet function and the limitation of platelet

thrombus infinite formation under pathophysiological flow conditions.

S6-2 Targeting Plasminogen Activator Inhibitor-1 to Prevent Vascular Disease Jian-Bo Wu Drug Discovery Research Center, Luzhou Medical College, Luzhou, Sichuan, China

Plasminogen activator inhibitor-1 (PAI-1) is the primary inhibitor of urinary-type and

tissue-type plasminogen activators and a key regulator of fibrinolysis. PAI-1 also regulates the

function of vascular cells, including vascular smooth muscle cells (VSMCs). PAI-1 inhibits

VSMC migration by inhibiting plasmin formation and preventing degradation of extracellular

matrix (ECM) and elastic laminae. PAI-1 binds vitronectin (VN), an ECM protein whose PAI-1

binding site overlaps with those on VN for integrin αVβ3 and u-PA receptor (uPAR), cell

surface proteins that control VSMC migration.

http://atvb.ahajournals.org/content/29/10/1565.long - ref-4

Therefore, PAI-1 can competitively block VSMC-VN interactions and inhibit migration.

However, PAI-1 can also promote VSMC migration by binding to uPAR-bound u-PA, leading

to conformational changes in PAI-1 and exposure of its high-affinity binding site for LDL

receptor-related protein (LRP). Binding of PAI-1 to LRP triggers internalization of PAI-1,

along with associated u-PA, uPAR, and integrin. This internalization process, which appears to

occur predominantly at the trailing edge of cells, allows VSMCs to detach from the ECM, a

process necessary for migration. PAI-1 has been shown to exhibit both pro- and anti-angiogenic

activities. Based on available data, it is hypothesized that the pro-angiogenic effect of PAI-1 is

mediated by stabilizing the ECM through inhibition of plasmin-mediated proteolysis, whereas

the anti-angiogenic effects of PAI-1 are mediated by inhibition of VN-dependent cell adhesion.

Therefore, that the pharmacologic inhibition of elevated PAI-1 might restore the impairments in

neovasculature observed in type II diabetes.

http://atvb.ahajournals.org/content/29/10/1565.long#ref-4

S6-3 Caffeic acid inhibits platelet activation ex vivo and thrombus formation in vivo Yu Lua,b,c,d,e, Quan Lib,c,d,e, Yu-Ying Liub,c,d,e, Kai Sunb,c,d,e, Lei Tub,d,e, Jing-Yu Fanb,c,d,e, Chuan-She Wangb, c,d,e * and Jing-Yan Hanb, c,d,e *

a Department of Gynecological, Beijing Royal Integrative medicine hospital

b Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China c Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China d Key Laboratory of Microcirculation, State Administration of Traditional Chinese Medicine, China e Key Laboratory of Stasis and Phlegm, State Administration of Traditional Chinese Medicine of the People's Republic of China, Beijing 100191, China. Caffeic acid (CA), one of the active constituents of Radix Salvia miltiorrhizae, exhibits antioxidant and anti-inflammatory activities. Platelet activation is relevant to a variety of acute thrombotic events and coronary heart diseases. There have been few studies on the effect of CA on platelets. Therefore, the aim of this study was to examine the ability of CA to modulate agonist-induced platelet activation in vivo and in vitro. The antithrombotic effect of CA was evaluated in mice using intravital microscopy in topical application of adenosine diphosphate (ADP)-induced cerebral venule thrombosis model. CA in vitro effects on agonist-induced platelet integrin-αIIbβ3 activation and granule-secretion were examined. Src, MAPK expressions and levels of tyrosine phosphorylation of the αIIbβ3 signaling pathway components were also studied. Our results demonstrated that CA (1-100 μM) dose-dependently inhibited thrombin- or ADP-induced αIIbβ3 activation and P-selectin expression in washed platelets, and it attenuated Src, ERK, p38MAPK and β3 chain cytoplasmic tail phosphorylation. Furthermore, CA (5 mg/kg) significantly inhibited in vivo thrombus formation without affecting tail bleeding time. Most importantly, this study demonstrated for the first time that CA possesses potent antiplatelet activity, which may be through modulation of αIIbβ3 signaling events, involving inhibition of Src, ERK and p38-MAPK. These data suggest that CA may have therapeutic potential for the treatment of cardiovascular or cerebrovascular diseases involving aberrant platelet activation.

S6-4

Trapping red blood cells within microvessels in living mice by optical

tweezers

Xun-Bin Wei

School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, China

This research is to explore the possibility of establishing an animal model for thrombosis. The

recent development of non-invasive imaging techniques has enabled the visualization of

molecular events underlying cellular processes in live cells. Although microscopic objects can

be readily manipulated at the cellular level, additional physiological insight is likely to be

gained by manipulation of cells in vivo, which has not been achieved so far. Here we use

infrared optical tweezers to trap and manipulate red blood cells within subdermal capillaries in

living mice. We realize a non-contact micro-operation that results in the clearing of a blocked

microvessel. Furthermore, we estimate the optical trap stiffness in the capillary. Our work

xpands the application of optical tweezers to the study of live cell dynamics in animals. e

Poster 1 Myocardial damage and microcirculation Chair: Chun-Shui Pan

Tasly Microcirculation Research Center, Peking University Health Science Center 13:20-13:30 P1-1 Effect of inoculated Walker256 Sarcoma on angiogenesis of myocardial tissue in rats with acute myocardial infarction Jian-Gang Liu Department of Cardiology, Xiyuan Hospital, China Academy of Chinese Medical Sciences 13:30-13:40 P1-2 The ameliorating effects of long term electroacupuncture at BaiHui (DU20) and ZuSanLi (ST36) on blood pressure and myocardial hypertrophy in spontaneous hypertension rat and underlying mechanisms Ze-Jun Huo School of Acupuncture and Moxibustion, Beijing University of Chinese Medicine 13:40-13:50 P1-3 The impact of rho kinase inhibition on cardiac and plasma angiotensin (1-7) in pressure-overload rats Hui Cai Department of Integrated Traditional and Western Medicine, Nanjing General Hospital of Nanjing Military Region PLA 13:50-14:00 P1-4 Effect of panax quinquefolius saponin on protein expression of ischemic myocardial osteopontin and tenascin-C in rats after myocardial Infarction Lei Zhang Cardiology Department of Xiyuan Hospital, China Academy of Chinese Medical Sciences 14:00-14:10 P1-5 Sang-qi Granula reduces blood pressure and myocardial fibrosis by suppressing inflammatory responses associated with the peroxisome proliferator-activated receptors and nuclear factor κB protein in spontaneously hypertensive rats Lan-Yu Chen

China-Japan Friendship Hospital 14:10-14:20 P1-6 QiShenYiQi Pills@, a compound Chinese medicine, ameliorates doxorubicin-induced myocardial structure damage and cardiac dysfunction in rats Dong-Xin Tang Key laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Integrated Traditional and Western Medicine, Peking University Cancer Hospital & Institute

P1-1 Effect of inoculated Walker256 Sarcoma on angiogenesis of myocardial tissue in rats with acute myocardial infarction Jian-Gang Liu, Da-Wu Zhang, Da-Zhuo Shi, Jie Li Department of Cardiology, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091 China; This work was supported by the National Natural Science Foundation of China (81173384) Objective: To establish the animal model of combined acute myocardial infarction (AMI) with tumor, and to observe angiogenesis and expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in tumor-bearing rat myocardium after inoculated Walker256 Sarcoma. Methods: Wistar rat’s left anterior descending coronary artery was ligated and the AMI model was established, taking the ascites of the rat with Walker256 Sarcoma by passage in seventh day and diluting ascites to 2×106/ml cell suspension. The survived rats after AMI were randomly divided into AMI model group (AMI group), postinfarction inoculated Walker256 Sarcoma Group (injected subcutaneously 0.2ml tumor cell suspension with aseptic method in right Regio axillaris of each rat and established into focal tumor model, referred to as AMI tumor-bearing group), sham operation and inoculated tumor group (the suture was placed inferior to the left anterior descending coronary artery without ligation, and inoculated tumor cell, referred to as sham tumor-bearing group) and sham control group (the suture was placed inferior to the left anterior descending coronary artery without ligation, and did not inoculate tumor cell, referred to as sham control group ), each group comprises 8 rats. Rats were sacrificed after 15 days, serum contents of VEGF, transforming growth factor ß1 (TGFß1) were examined by ELISA, the changes of angiogenesis in rat myocardial tissue were observed by immunohistochemistry, the expression of VEGF and bFGF in rat myocardium were evaluated by immunohistochemistry, and the changes of microvessel density in the tumor tissue and ischemic myocardium were detected by optical microscope. Results: ① Compared with sham control group, the serum contents of VEGF and TGFß1 increased significantly in AMI tumor-bearing group and sham tumor-bearing group (P<0.01), which were significantly higher than that in AMI group (P<0.01). ② Compared with sham control group, the expression of VEGF and bFGF increased significantly in AMI tumor-bearing group and AMI group (P<0.05, P<0.01), and microvessel density in the ischemic myocardium were also higher (P<0.05, P<0.01). ③ Compared with AMI group, the expression of VEGF in ischemic myocardium of AMI tumor-bearing rats increased significantly (P<0.01), and microvessel density in the ischemic myocardium were also higher (P<0.05). Conclusion: after inoculated walker256 sarcoma on established AMI rat model, VEGF contents in the serum and ischemic myocardium increased, and microvessel density increased. The mechanism may be related to the effect of angiogenesis factor on the ischemic myocardium.

P1-2 The ameliorating effects of Long term electroacupuncture at BaiHui (DU20) and ZuSanLi (ST36) on blood pressure and myocardial hypertrophy in spontaneous hypertension rat and underlying mechanisms Ze-Jun Huoa,b,c, Quan Lib, Gui-Hua Tiana,b, Chang-Man Zhoud, Xiao-Hong Weib, Chun-Shui Panb, Lei Yangd, Yan Baie, You-Yi Zhange, Ke Heb, Zhi-Jun Lia and Jing-Yan Hana,f a School of Acupuncture and Moxibustion, Beijing University of Chinese Medicine, Beijing, China b Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China c Department of Traditional Chinese Medicine, Peking University Third Hospital, Beijing, China d Department of Anatomy, School of Basic Medical Sciences, Peking University, Beijing, China e Institute of Vascular Medicine, Peking University Third Hospital and Key Laboratory of Molecular Cardiovascular Science, Chinese Ministry of Education, Beijing, China f Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China The purpose of this study is to investigate the inhibitory effects of long term electroacupuncture stimulation at BaiHui (DU20) and ZuSanLi (ST36) on blood pressure, myocardial hypertrophy and myocardial fibrosis of spontaneous hypertension rat (SHR) and underlying mechanisms. 6-weeks-old SHR or Wistar male rats were randomly divided into 6 groups, the control group (SHR/Wistar), the non-acupoint electroacupuncture stimulation group (SHR-NAP/Wistar-NAP) and the electroacupuncture stimulation at DU20 and ST36 group (SHR-AP/Wistar-AP), with 24 rats in each group. For SHR-AP/Wistar-AP group and SHR-NAP/Wistar-NAP group, the penetrating accorded to the Chinese rat acupoint positioning standard. For the control groups, the rats only were binding up without acupuncture and electrical stimulation. Blood pressure was determined before and at 2, 4, 6, and 8 weeks after electroacupuncture treatment by the tail-cuff method. At the end of 8th weekend, the ventricular thickness and heart function of the rat were detected utilizing echocardiography. The content of angiotensin II (Ang II), endothelin-1 (ET-1) and NO in the plasma were determined using enzyme-linked immunosorbent assay. Histological studies were also performed on the heart and the ascending aorta. The expression of angiotensin II type 1 receptor (AT1R), endothelin-1 type A receptor (ETAR), eNOS and iNOS in rat myocardium and ascending aorta were investigated by the western blotting method. The results demonstrated that mean arterial pressure, aortic wall and ventricular thickness and heart/body weight ratio of SHR were significantly reduced with long term electroacupuncture stimulation at DU20 and ST36. The content of Ang II, ET-1 in SHR plasma decreased while the content of NO increased after electroacupuncture treatment. Long term electroacupuncture significantly inhibited protein expression of AT1R, ETAR and iNOS in rat myocardium and ascending aorta, while enhanced the eNOS expression in SHR. The beneficial effects of long term electroacupuncture stimulation at DU20 and ST36 on heart function may be related to decrease the content of Ang II and ET-1 while increase the content of NO in peripheral blood, and inhibit the protein expression of AT1R, ETAR, iNOS in cardiac muscle and aortic wall while enhance the expression of eNOS.

P1-3

The Impact of Rho kinase inhibition on cardiac and plasma Angiotensin (1-7) in pressure-overload Rats

Hui Cai, Pei-Yong Zhang, Xiao-Lei Dong, Ling-Jie Zhao, Zhi-Ming Zhao Department of Integrated Traditional and Western Medicine, Nanjing General Hospital of Nanjing Military Region PLA, Nanjing 210002, Jiangsu China Objective: Inhibition of Rho-kinase displays property against cardiac remodeling, although its

effect on has not been fully elucidated. The present study was designed to investigate the

impact of fasudil, a Rho kinase inhibitor, on cardiac and plasma AngiotensinⅡ (AngⅡ) and

Angiotensin (1-7) (Ang (1-7)) in pressure-overload Rats.

Methods: The SD rats were prepared with abdominal aorta constriction and fed under standard

condition. 4 weeks after operation, these rats were divided into 4 groups randomly: Sham

group, Model group, Fasudil High-dose group (FH, 30 mg/kg⋅d) and Fasudil Low-dose group

(FL, 10mg/kg⋅d). At 8 weeks after surgery, the Heart Weight/Body Weight Index (HWI) and

Left Ventricular Weight Index (LVWI) were calculated, and the content of cardiac

hydroxyproline (HYP) was measured using alkaline hydrolysis. Study the myocardial

histopathology and interstitial fibrosis with HE and Masson staining. Cardiac and plasma

AngⅡ and Ang (1-7) concentration were detected with ELISA.

Results: Comparing with Sham group, the LVWI and HYP content were significantly elevated

in model group (p<0.01). Much more collagen deposited in myocardial interstitium, associated

with increased cardiac and plasma AngⅡ. The levels of cardiac Ang (1-7) were reduced

(p<0.01), but plasma Ang (1-7) was similar to Sham group. Comparing with Model group,

Fasudil reduced the pressure-overload mediated elevation of LVMI and HYP content, and

reduced interstitial collagen deposition. Cardiac and plasma AngⅡ protein in FH group were

normalized. Moreover, cardiac and plasma Ang (1-7) were increased significantly (p<0.01). In

the FL group, cardiac and plasma AngⅡ protein were reduced (p<0.05), associated with Ang

(1-7) increased (p<0.01).

Conclusion: These results indicate that Fasudil can attenuates pressure-overload mediated

cardiac fibrosis, associated with reduction of AngⅡ and increased Ang (1-7) levels.

Key Words: Fasudil; Cardiac Remodeling; Cardiac Fibrosis; Renin-Angiotensin System.

P1-4

Effect of panax quinquefolius saponin on protein expression of ischemic myocardial osteopontin and tenascin-C in rats after myocardial Infarction

Lei Zhang, Qing-Xiang Zhang, Jian-Gang Liu, Guo-Ju Dong, Da-Zhuo Shi Cardiology Department of Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091, China Aim To investigate the effect of panax quinquefolius saponin (PQS) on protein expression of ischemic myocardial osteopontin (OPN), tenascin-C (TN-C) and heart function during prophase ventricular remodeling (VR) after acute myocardial infarction (AMI) in rats. Meanwhile, explore the possible mechanism on extracellular matrix (ECM) metabolism. Methods The AMI model was established by ligating the left anterior descending coronary artery of rats to induce myocardial infarction. 60 Wistar rats with model establishment were randomly divided into 4 groups, 15 rats in each group. They were sham group (the suture was penetrated around the left anterior descending coronary artery, but not tied), model group, valsartan group of which valsartan was given 7.2mg/(kg·d), PQS group of which panas quinquefolium saponin was given 162mg/(kg·d). All of the medicine was given by gavage from the second day after model establishment. Both sham and model group were administered of equivalent distilled water. After continuous intragastric administration of 4 weeks, ultrasonic cardiography was used to examine the morphologic change and heart function. HE staining was used to observe the myocardial pathological change. The protein expression of OPN and TN-C in myocardial tissue was tested semiquantitatively by immunohistochemical method. Myocardium contents of angiotensinⅡ (AngⅡ), aldosterone (ALD) and tissue growth factor-β1 (TGF-β1) were tested by radioimmunoassay method. Results Compared with model group: (1) 1eft ventricular end-systole dimension (LVED) reduced significantly and left ventricular ejection fraction (LVEF) increased significantly in PQS group (P<0.05); (2) both interventricular septum end-diastole thickness (IVSTd) and interventricular septum end-systole thickness (IVSTs) increased significantly in PQS group (P<0.05); (3) ischemic myocardium protein expression of OPN decreased significantly in PQS group (P＜0.01) and TN-C decreased significantly in PQS group (P<0.05); (4) myocardial tissue contents of AngⅡ, ALD and TGF-β1 decreased significantly in PQS group (P<0.05, P<0.01). Conclusion PQS can improve the heart function by attenuating the prophase VR of rats after AMI. The possible mechanism of the improvement is that PQS can regulate ECM metabolism by inhibiting neuroendocrine factors production, alleviating inflammation reaction and reducing ischemic myocardial protein expression of OPN and TN-C.

http://dict.cnki.net/dict_result.aspx?searchword=%e7%ad%89%e9%87%8f&tjType=sentence&style=&t=equivalent

http://dict.cnki.net/dict_result.aspx?searchword=%e5%85%8d%e7%96%ab%e7%bb%84%e5%8c%96%e6%b3%95&tjType=sentence&style=&t=immunohistochemical+method

P1-5

Sang-qi Granula Reduces Blood Pressure and Myocardial Fibrosis by Suppressing Inflammatory Responses Associated With the Peroxisome Proliferator-Activated Receptors and Nuclear Factor κB Protein in Spontaneously Hypertensive Rats Lan-Yu Chena , Chun-Shui Panb , Xiao-Hong Weib , Lin Lia , Jing-Yan Hanb , Li Huanga

a China-Japan Friendship Hospital b Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China Aim: Sang-qi Granula (SQ) is a compound preparation of Chinese medcine and currently used for treatment of hypertension in China. Given its protective effects on cardial function in decreasing blood pressure, we investigated the mechanism of protective effects of SQ on myocardium. Methods: 16 male normal Wistar-Kyoto rats and 16 Spontaneous Hypertension Rats (SHR) were employed without medical treatment. 16 SHR were employed with SQ treatment. Rats in each group were sacrificed at two time points (8 week-treatment and 16 week- treatment). Blood pressure (BP), Heart Weight/Body Weight (HW/BW) were measured. The expression of myeloperoxidase (MPO), ICAM-1, TNF-α and CD68-positive cells were assessed. The interstitial collagen volume fraction (CVF) and perivascular collagen volume area (PVCA), the expression of TGF-β, Smad-3 and PPARα, γ, NF-κB (P65 and P50) were observed. Results: SQ significantly inhibited the elevation of the blood pressure and HW/BW of SHR. Next, SQ prevented myocardial fibrosis. Finally, pro-inflammatory mediator associated with NF-κB (TNF-α, ICAM-1, MPO, CD68) and TGF-β, Smad-3 related to collagen deposition, which is upregulated in SHR group, was significantly suppressed by SQ. Expression of NF-κB and was decreased in SHQ+SQ group compared to PPARα, γ expression was increased by SQ. Conclusion: Treatment with SQ ameliorates cardial fibrosis induced by hypertension by attenuating the upregulation of ICAM-1, TNF-α, MPO, TGF-β, Smad-3, P65 and P50 expression and improving PPARα and PPARγ expression level. The results suggest that SQ may be an option for preventing cardial fibrosis through PPAR signal pathway. Key Words: hypertension, myocardial fibrosis, PPAR, NF-kappaB, inflammation

P1-6 QiShenYiQi Pills@, a Compound Chinese Medicine, Ameliorates Doxorubicin-Induced Myocardial Structure Damage and Cardiac Dysfunction in Rats

Dong-Xin Tang1,3,4, Hai-Ping Zhao3, Chun-Shui Pan3, Yu-Ying Liu3, Xiao-Hong Wei3, Xiao-Yuan Yang2,3, Yuan-Yuan Chen2,3, Jing-Yu Fan3, Chuan-She Wang2,3, Jing-Yan Han2,3, Ping-Ping Li1

1 Key laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Integrated Traditional and Western Medicine, Peking University Cancer Hospital & Institute, Beijing 100142, China 2 Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing 100191, China

3 Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing 100191, China 4 Department of Oncology, The First Affiliated Hospital of Guiyang College of TCM, Guiyang 550002, China QiShenYiQi Pills® (QSYQ) is a compound Chinese medicine used for treatment of cardiovascular diseases. The present study investigated the effects of QSYQ on the doxorubicin (DOX)-induced disorders in rat cardiac structure and function, and the possible mechanism underlying. A total of 24 male Sprague-Dawley rats were administrated by intraperitoneal injections with DOX at a dose of 2.5 mg/kg, once every other day for a total of 6 times. After the 6th injection, the rats were evaluated by echocardiographic analysis, and the animals with injured heart (n=14) were divided into 2 groups and further treated with (n=7) or without (n=7) QSYQ by gavage at a dose of 0.2g/day, once a day, over the next 2 weeks. Two weeks after QSYQ treatment, the following variables were assessed: myocardial blood flow (MBF) by Laser-Doppler Perfusion Imager, the ratio of heart weight to body weight (HW/BW), myocardial histology, myocardial content of ATP, AMP, free fatty acids (FFA) and AMP/ATP by ELISA, and expression of PPAR α, PGC-1 α, ATP 5D by Western blot. Statistical analysis was performed using one-way ANOVA followed by Turkey test for multiple comparisons. DOX challenge significantly increased left ventricular internal diameter and HW/BW, decreased the thickness of left ventricular posterior wall, left ventricle ejection fraction, and the left ventricle fractional shortening. DOX also increased AMP, FFA, AMP/ATP and decreased ATP, down-regulated the protein content of ATP 5D, PPAR α and PGC-1 α. All these DOX-induced cardiac insults were attenuated significantly by QSYQ treatment. These results show the potential of QSYQ to ameliorate DOX-induced disorders in cardiac structure and function, this effect may be related to the increase in myocardial ATP content via up-regulation of ATP 5D, PPAR α and PGC-1 α, and oxidation of FFA.

Poster 2 Pharmacology of Chinese medicine for microcirculatory disturbance

Chair: Fei Ye

Beijing Key Laboratory of New Drug Mechanism of Action and Pharmacodynamic

Evaluation, Chinese Academy of Medical Sciences, Beijing Union Medical College,

Institute of Meteria Medica

13:20-13:30 P2-1 QiShenYiQi Pills® prevent cardiac ischemia-reperfusion injury via energy modulation Se-Qi Lin Key Laboratory of Modern Preparation of Traditional Chinese Medicine, Ministry of Education, Jiangxi University of Traditional Chinese Medicine 13:30-13:40 P2-2 Effects of Guanxinkang on blood lipids, blood rheology, atheromatous plaque and related expression in atherosclerosis of apoE-/- mice He-Wei Qin Longhua Hospital Affiliated to Shanghai University of TCM, Shanghai 200032, China 13:40-13:50 P2-3 Observation about time-effect relation of extracts from ginseng notoginseng and chuanxiong on vascular endothelial cells senescence Ming Wang The Cardiology Department of Chongqing Hospital of Traditional Chinese Medicine and Chongqing the First Hospital 13:50-14:00 P2-4 HEB improves the functional damage of gastric smooth muscle in diabetic rats Zhang-Sen Hao School of Pharmacy, Hebei Medical University, Shijiazhuang, China 14:00-14:10 P2-5 Panax notogiseng saponins ameliorates rats mesenteric microcirculatory disturbance induced by I/R Yu Zhang Tasly Microcirculation Research Center, Peking University Health Science Center 14:10-14:20 P2-6 The studies of Xuefuzhuyu oral liquid intervention on the SIRT1 signal transduction mechanism in Ischemic cardiomyocyte apoptosis Kui-Wu Yao Guang’anmen Hospital, China Academy of Chinese Medical Sciences

P2-1 QiShenYiQi Pills® Prevent Cardiac Ischemia-reperfusion Injury via Energy Modulation Se-Qi Lin a, b, c, Xiao-Hong Wei c, Ping Huang c, Yu-Ying Liu c, Na Zhao c, Quan Li c, Chun-Shui Pan c, Bai-He Hu c, Xin Chang c, Jing-Yu Fan c, Xiao-Yuan Yang c, Chuan-She Wang

c, d, Hong-Ning Liu a*, Jing-Yan Han c, d*

aKey Laboratory of Modern Preparation of Traditional Chinese Medicine, Ministry of Education, Jiangxi University of Traditional Chinese Medicine, Nanchang, China bGraduate School, Hunan University of Chinese Medicine, Changsha, China cTasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China dDepartment of Integration of Traditional Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China

Background: QiShenYiQi Pills® (QSYQ) is a compound Chinese medicine used in China for

alleviating cardiac function. The present study was designed to explore the effect and

mechanism of QSYQ on ischemia-reperfusion (I/R)-induced disorders in myocardial structure

and function, with particularly focusing on the regulation of energy metabolism.

Methods: Sprague-Dawley rats, with or without QSYQ pretreatment, were subjected to 30 min

occlusion of the left anterior descending coronary artery and followed by 90 min reperfusion.

Myocardial blood flow (MBF), and cardiac function were evaluated at baseline, immediately

after ischemia and 30, 60, 90 min after reperfusion. Ninety minutes after reperfusion,

myocardial infarction, myocardial histology and ultrastructure were assessed. Terminal

deoxynucleotidyl transferase-mediated dUTP nick end labeling staining was conducted to

assess myocardial cell apoptosis. ATP, ADP and AMP content was determined by

Enzyme-Linked Immunosorbent Assay, F-actin in myocardial cells determined by

immunofluorescence microscopy, and expression of ATP synthase α, ATP5D, and

phosphorylated-Myosin Light Chain (P-MLC) determined by western blotting.

Results: Pre-treatment with QSYQ protected against I/R-induced MBF decrease, myocardial

infarct and apoptosis, and cardiac dysfunction. Moreover, I/R caused an increase in P-MLC,

meanwhile, an alteration in the ratio of ADP/ATP and AMP/ATP, and a reduction in expression

of ATP 5D, all of which were significantly ameliorated by pre-treatment with QSYQ.

Conclusions: The results of the present study suggest an involvement of regulation of energy

metabolism in the action of QSYQ to protect against myocardial I/R injury.

P2-2 Effects of Guanxinkang on Blood Lipids, Blood Rheology, Atheromatous Plaque and related Expression in Atherosclerosis of ApoE-/- mice He-Wei Qin, Ping Liu Longhua Hospital Affiliated to Shanghai University of TCM, Shanghai 200032, China Objective: To observe the effects of Guanxinkang on blood lipids, blood rheology atheromatous plaque and related expression in atherosclerosis of ApoE-/-mice, explore the mechanism of Guanxinkang prevent hyperlipidemia, atherosclerosis and coronary heart disease. Methods: ApoE-/- mice 50, C57BL/6J mice 10 as the normal group, SPF grade, male and female, 6 weeks old, body weight (18±2) g. All animals were housed in the Court of SPF animal center. C57BL/6J mice were given a normal diet, ApoE-/- mice were given a high fat diet. Each group shall be free access to water. To 12 weeks of age, the ApoE-/- mice were randomly divided into 5 groups according to the grouping method,the normal group and model group were given an equal volume of saline; Guanxinkang low, medium and high dose groups intragastric administration Guanxinkang 21.6, 43.2, 86.4 g / (kg·d); simvastatin group intragastric administration of simvastatin 3mg /(kg·d). During the experiment, according to the body weight-adjusted dose. Fasting before animals were sacrificed 12h, after orbital blood, the serum lipid levels, rheology was measured. Isolated aorta, Undergoing aortic pathology observed damage and the use of immunohistochemical staining for lesions of monocyte chemoattractant protein-l, nuclear factor-κB content. Results: 1. The effects of Guanxinkang on ApoE-/- mice lipid metabolism: (1) Mice compared with the normal group, model group, serum TC, TAG, LDL-C and ApoB levels increased, HDL-C, ApoAⅠ and ApoE content decreased, the difference was statistically significant (P<0.05). (2) Compared with model group, the treatment group serum TC, TAG, LDL-C and ApoB decreased, HDL-C and ApoA Ⅰ content increased, the difference was statistically significant (P<0.05). (3) High-dose group and middle dose Guanxinkang group, serum TAG content decreased, HDL-C levels increased (P<0.05), serum TC, LDL-C, ApoB and ApoAⅠ was no significant difference (P>0.05); Guanxinkang high dose group and low dose Guanxinkang group, serum HDL-C and ApoAⅠ levels rise, TAG, TC and ApoB levels decreased, the difference was statistically significant (P<0.05). Guanxinkang middle dose group and low dose Guanxinkang group, serum TC content decreased, TAG and ApoAⅠ content increased, the difference was statistically significant (P<0.05), serum LDL-C, ApoB, and HDL-C, the difference not statistically significant (P> 0.05). Compared with simvastatin group and high dose Guanxinkang serum HDL-C and ApoA Ⅰ levels rise, TAG, TC and ApoB levels decreased, the difference was statistically significant (P<0.05); middle dose Guanxinkang serum HDL-C content increased, TC and ApoB levels decreased, the difference was statistically significant (P<0.05); low dose Guanxinkang serum HDL-C levels rise, ApoB content decreased, the difference was statistically significant (P<0.05). (4) Treatment group and model group between the ApoB content pairwise comparisons, the difference was not statistically significant. 2 .The effect of Guanxinkang on ApoE-/- mice hemorheology indexes: ApoE-/- mice showed blood viscosity elevated state, and the rats Guanxinkang on whole blood viscosity and plasma viscosity indicators compared with the model group were significantly improved, (P<0. 01); and simvastatin group, the difference was significant. 3. The effects of Guanxinkang on atherosclerotic plaques of ApoE-/- mice : Compared with model group, the treatment group luminal area (LA) large intimal thickness (IMT) thin plaque area (PA) is small, fibromuscular component (FS) less lipid center area (CA) is small, the minimum thickness of the fibrous cap (FCT) thin plaque area and lumen area ratio (PA /LA) is small, central area of lipid plaque area ratio (CA/PA) is small, the center area and the fiber lipid muscular component ratio (CA/FS) Large. 4. The effects of Guanxinkang on Atheromatous Plaque and Peripheral vascular wall MCP-1, NF-κB Expression in Atherosclerosis of ApoE-/-

mice: MCP-1 and NF-κB mean optical density value is small (P<0.05). Guanxinkang high dose group was superior to simvastatin group (P<0.05). Guanxinkang low, medium and high dose groups compared between the dose-effect relationship does not exist, but the most obvious effect is the high dose group of Guanxinkang. Conclusion: Guanxinkang can regulate fat diet-induced lipid metabolism disorders and abnormal blood rheology changes, delay ApoE-/-mice atherosclerotic lesions, reduce expression of MCP-1 and NF-�B; this study showed that mechanism of Guanxinkang prevented hyperlipidemia, atherosclerosis and coronary heart disease Associated with these.

P2-3 Observation about time-effect relation of extracts from ginseng notoginseng and chuanxiong on vascular endothelial cells senescence Ming Wang 1, Yan Lei 2

Supported by National Natural Science Foundation of China (No. 30973828); Major State Basic Research Development Program of China (937 Program, No. 2007CB507400); Self-Project Foundation from China Academy of Chinese Medical Sciences (No. Z02151) Correspondence to: Prof. LEI Yan, E-mail: [email protected]. The Cardiology Department of Chongqing Hospital of Traditional Chinese Medicine and Chongqing the First Hospital, 400021 2. The Experimental Research Center of Chinese Academy of Chinese Medical Science, 100700

Objective: To observe the time-effect relation of extracts from ginseng notoginseng and chuanxiong on angiotensin Ⅱ-induced senescence of vascular endothelial cells and explore the feature of Chinese medicine against vascular diseases. Methods: Human umbilical vein endothelial cells (HUVECs) cultured in vitro were stimulated with 10-6mol/L AngⅡ to induce cell senescence, which were divided into 4 groups, the blank control group, the AngⅡ model group, the extracts group and the Telmisartan group. The β-gal was used to identify senescence of cells, the CCK-8 method was applied to assess the cell viability, the cell function was examined with the level of eNOS and the flow cytometry was used for analyzing the cell cycle changes. Results: Compared with the control cells, the cells positive for β-gal staining was significantly increased in AngⅡ model group, and showed cell cycle arrest at G0/G1 phase with decreased S and G2/M phase cell percentage, eNOS expression and cell viability (p＜0.05). The extracts and Telmisartan treatment of AngⅡ-induced cells resulted in decreased β-gal positive cells with a reduction in G0/G1 phase cells and an increasing in S, G2/M phase cells and eNOS expression (p＜0.05). At 24h, the extracts were more effective than Telmisartan (p＜0.05); while Telmisartan was more effective at 48h (p＜0.05). Conclusion: Extracts from ginseng notoginseng and chuanxiong can delay AngⅡ-induced aging of HUVECs and may play an important role in early senescence. KEYWORDS extracts from ginseng notoginseng and chuanxiong, angiotensin Ⅱ, vascular

endothelial cells, senescence

P2-4

HEB improves the functional damage of gastric smooth muscle in diabetic rats Zhang-Sen Hao, Ding Zhao School of Pharmacy, Hebei Medical University, Shijiazhuang, China Objective: To investigate the effects of HEB (a compound abstracted from a traditional Chinese medicine) on impaired gastric smooth muscle in diabetic rats. Methods: The gastric fundus and pylorus strips were isolated from streptozotocin-induced diabetic rats. The contractions and relaxations were induced by electrical field stimulation (EFS), and the changes of those reactions on gastric smooth muscle in diabetic rats were measured. Neurogenic reactions induced by EFS were confirmed by blocking action potentials in nerves with 0.1 µmol/L TTX. In contractile experiments, electrical impulses (50 V, 20 Hz, 1 ms for 10 s, train duration with 100-s intervals between the trains) were provided by a modified stimulator. HEB 1, 10 µmol/L or atropine 1 µmol/L were added to the organ bath to test the effects of HEB on the contractile responses to EFS in the rat isolated gastric fundus strips. In relaxant experiments, gastric fundus strips were pre-contracted with 0.1 μmol/L substance P (SP), and the relaxation response to transmural stimulation (50 V, 1-10 Hz, 1 ms for 5 s, train duration with 100-s intervals between the trains) was studied once the contractile response to SP had reached its plateau; the relaxation response of pyloric strips was studied in the same parameters of stimulation with gastric fundus strips at resting tension. HEB 1, 10 µmol/L or L-NAME 100 μmol/L, suramin 100 μmol/L was added to the organ bath to test the effects of HEB on the relaxation responses to EFS in isolated gastric fundus and pyloric strips. Results: EFS elicited reproducible contractions and relaxations in isolated gastric fundus and pylorus strips from both normal and diabetic rats. In the diabetic rats, the EFS-induced contractile response was decreased and disordered, EFS-induced relaxant response was increased on gastric fundus; but the EFS-induced relaxant response on pylorus strips was decreased, as compared with normal rats. All the responses induced by EFS were abolished by TTX. HEB significantly increased EFS-induced contractile response and improved disordered contraction by adjusting the neurotransmitter release on cholinergic nerve, but it had no obvious effect on the relaxation response on gastric fundus strips in diabetic rats; however, HEB inhibited EFS-induced relaxation response on pylorus strips in diabetic rats. Conclusion: HEB improves the functional damage of gastric fundus smooth muscle by affecting the neurotransmitter release on cholinergic nerve in diabetic rats.

P2-5 Panax notogiseng saponins Ameliorates rats mesenteric microcirculatory disturbance induced by I/R Yu Zhang1,2, Kai Sun1, Yu-Ying Liu1, Chun-Sui Pan1, Ke He1,2, Ming-Xia Wang1 and Jing-Yan Han1,2 1) Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing,

China 2) Department of Integration of Chinese and Western Medicine, School of Basic Medical

Sciences, Peking University, Beijing, China

AIMS: To investigate whether PNS has beneficial effects on microcirculatory disturbance induced by superior mesenteric artery ischemia and reperfusion, and the possible mechanism. METHODS: After anesthetized, male Wistar rats (200-250 g) ligating the superior mesenteric artery and vein for 10 min following reperfusion for 60 min to establish the microcirculatory disturbance model. Continuously infusion with PNS (5 mg/kg/hr) via left jugular vein, from 10 min before ischemia or 10 min after reperfusion, until 60 min after reperfusion. In vivo dynamics of endothelial peroxidation, leukocytes adhesion, and albumin leakage of rats mesenteric venules was observed by a CCD camera or a silicon intensified target camera connected to an inverted intravital microscopy. Mast cell degranulation was examined by 0.1% toluidine blue dying, 60 min after reperfusion. Serum MPO level was evaluated by ELISA. Intestinal ICAM-1 expression and Src phosphorylation were detected by western blotting. RESULTS: Superior mesenteric artery I/R lead to endothelial peroxidation, leukocyte adhesion, albumin leakage and interstitial mast cell degranulation of rats mesentery venule. Both pre- and post-administration of PNS ameliorated leukocyte adhesion and mast cell degranulation, while with no obvious effects to endothelial peroxidation and albumin leakage. In addition, PNS inhibited the serum MPO increase and intestinal ICAM-1 expression and Src phosphorylation induced by I/R. CONCLUSIONS: PNS ameliorated I/R induced leukocyte adhesion and interstitial mast cell degranulation of rat mesenteric venule. The inhibition of leukocyte adhesion of PNS is related to its inhibition of Src phosphorylation and ICAM-1 expression.

P2-6

The studies of Xuefuzhuyu oral liquid intervention on the SIRT1 signal transduction mechanism in Ischemic cardiomyocyte apoptosis Kui-Wu Yao, Zhang-Jing Liu Guang’anmen Hospital, China Academy of Chinese Medical Sciences Objective: To observe the morphological changes of H9c2 rat myocardial model cells under light microscope following intervention with Xuefuzhuyu oral liquid (XFZY), Oxygen-glucose deprivation (OGD) and silent information regulator 2-related enzymes 1 (SIRT1), as well as changes of Sirt1 pathway related genes (FOXO family, NF-kB, p53 etc) and cell activities, so as to clarify the gene target and corresponding mechanism of XFZY in the treatment of IHD under the TCM theory of eliminating blood stasis and promoting tissue regeneration. Method: H9c2 rat myocardial cells were divided into six groups: Normal group, OGD model group, SIRT1 transfection group, OGD+SIRT1 transfection group, OGD+Xuefuzhuyu oral liquid group, OGD+SIRT1 transfection+XFZY group. Cell counting kit-8 (CCK-8) was used to draw cell growth curve. The changes of cell morphology were observed with light microscope by the acridine orange (AO)/ethidium bromide (EB) doubly staining. The concentration of SIRT1 pathway related-genes and corresponding protein expression were tested by Real-Time PCR (RT-PCR) and Western-blot status after XFZY intervention and SIRT1 transfection. Results: 1. Result: The cells were observed after staining by light microscope. There was significant difference of cellular morphology among the 6 different groups. The chromatins exhibited green and normal structure in normal group and SIRT1 transfection group. The chromatins exhibited green and condensate in OGD model group .The chromatins were jacinth and condensate in OGD+SIRT1 transfection group. OGD+XFZY group showed that: the chromatins were green and condensate. The colour was lighter than OGD model group on cytomembrane. OGD+SIRT1 transfection+XFZY group showed that: the chromatins were lighter jacinth green and condensate. The colour was lighter than OGD+SIRT1 transfection group on cytomembrane. 2. RT-PCR results showed that the expression of SIRT1 mRNA decreased in OGD model group, SIRT1 transfection group, OGD+ SIRT1 transfection group, OGD+XFZY group, OGD+SIRT1 transfection+XFZY group, but increased after intervention; The expression of SIRT1 mRNA get similar in Normal group and OGD+XFZY group, but showed significant differences between Normal group and OGD+SIRT1 transfection+ XFZY group. The mRNA and corresponding protein expression of P53, NF-kB, FOXO1, FOXO3, FOXO4 increased significantly in OGD model group, SIRT1 transfection group, OGD+SIRT1 transfection group, OGD+XFZY group, OGD+SIRT1 transfection + XFZY group. 3. Western blot results showed that the expression of SIRT1 mRNA decreased in model group, but increased after intervention; mRNA and corresponding protein expression of p53, NF-kB, FOXO1, FOXO3, FOXO4 increased in model group；Protein expression of SIRT1, p53, NF-kB, FOXO1, FOXO3, FOXO4 showed significant differences (p<0.05). Conclusion: Our study proves that SIRT1 signal pathway has antiapoptotic effects in vitro OGD rat myocardial cells. SIRT1 signal pathway might be one of the targets of XFZY in anti-apotosis by modulating functions of its substrates and downstream genes. This study promotes the application research of XFZY in cardiac regeneration medicine and provides a clear target for ischemic heart disease treatment. Keywords: H9c2 rat myocardial cells; Ischemic heart diseases; Removing blood stasis for promoting tissue regeneration; SIRT1; Regulation mechanism.

Poster 3 Platelets

Chair: Yu-Ying Liu


13:20-13:30 P3-1 Arsenic trioxide induces platelet apoptosis Yi-Cun Wu

Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Jiangsu Institute of Hematology 13:30-13:40 P3-2 Effect of caffeic acid on mouse cerebral venular thrombosis induced by photochemical reaction Yu Lu Department of Gynecological, Beijing Royal Integrative Medicine Hospital 13:40-13:50 P3-3 Platelet apoptosis in uremic patients Ming Li The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology 13:50-14:00 P3-4 Surface expressions of platelet glycoprotein Ibα, GpⅡb/Ⅲa, and P-Selectin are elevated in lung cancer patients Jian-Sheng Zhang The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology 14:00-14:10 P3-5 miR30c regulates plasminogen activator inhibitor-1 in platelet Mao Luo

Drug Discovery Research Center, Luzhou Medical College 14:10-14:20 P3-6 Aspirin induces platelet apoptosis Li-Li Zhao School of Biological Science and Medical Engineering, Beijing University of Aeronautics and Astronautics

P3-1 Arsenic Trioxide Induces Platelet Apoptosis Yi-Cun Wu1, Jin Dai2, Wei-Lin Zhang1, Rong Yan1, Chang-Geng Ruan1 and Ke-Sheng Dai1*

1Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Jiangsu Institute of Hematology, Suzhou, China; 2School of life sciences, Peking University, Beijing, China *Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: 0086-512-67781370; Fax: 0086-512-67781370 [Objective] Arsenic trioxide, a component of Traditional Chinese Medicine, is known as an effective anticancer drug especially in the treatment of acute promyelocytic leukaemia (APL). APL has emerged as the most curable subtype of acute myeloid leukaemia since the widely use of arsenic trioxide-based chemotherapy. However, recent researches show that thrombocytopenia occurred in 79% of the relapsed or refractory APL patients treated with arsenic trioxide, and part of the APL patients had to be stopped treatment because of catastrophic bleeding, such as intracranial and pulmonary haemorrhage. Thrombocytopenia also occurred in 43% of the myelodysplastic syndrome patients treated with arsenic trioxide. Recently, arsenic trioxide has been proved to have a pro-apoptotic effect on various kinds of nucleated tumour cells or non-tumour cells. The effect of arsenic trioxide on enucleated platelet, however, still remains unclear. The aim of current study is to investigate whether arsenic trioxide induces platelet apoptosis. [Methods] Washed platelets (3 × 108/ml) were incubated with different concentrations of arsenic trioxide or vehicle at 37°C for 4 hours. Then, mitochondrial inner transmembrane potential (ΔΨm) and phosphatidylserine (PS) exposure were tested by flow cytometry. In the mean time, the treated platelets were analyzed by western blot for the expression levels of pro-apoptotic protein (Bax), and anti-apoptotic proteins (Bcl-2 and Bcl-XL). Activation of caspase-3 was also examined by western blot using an anti-caspase-3 antibody. [Results] ΔΨ depolarization and PS exposure were dose-dependently induced in platelets incubated with different concentrations (2 μM, 4 μM, 8 μM, 16 μM) of arsenic trioxide as detected by flow cytometry, and the lowest concentration of arsenic trioxide incurring ΔΨm depolarization and PS exposure was 4 μmol/L. Simultaneously, arsenic trioxide induced up-regulation of Bax and down-regulation of Bcl-2 and Bcl-XL in a dose-dependent manner. Furthermore, 17 kD cleaved caspase-3 fragments were dose-dependently induced in platelets treated with different concentrations of arsenic trioxide indicating that caspase-3 was activated by arsenic trioxide. [Conclusions] Taken together, the data indicate that arsenic trioxide induces platelet apoptosis in vitro, which might suggest a novel pathogenic mechanism of thrombocytopenia in the patients who treated with arsenic trioxide.

P3-2

Effect of Caffeic Acid on Mouse Cerebral Venular Thrombosis Induced

by Photochemical Reaction

Yu Lua,b,c, Quan Lib, Yu-Ying Liub, Kai Sunb, Chun-Shui Panb, Ke Heb, Xiao-Hong Weib,

Jing-Yu Fanb, Chuan-She Wanga,b* and Jing-Yan Hana,b*

a Department of Integration of Chinese and Western Medicine, School of Basic Medical

Sciences, Peking University, Beijing, China b Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing,

China c Department of Gynecological, Beijing Royal Integrative Medicine Hospital

The purpose of this study was to investigate the effect of caffeic acid (CA) on mouse cerebral

venular thrombosis induced by photochemical stimulation (PS). Photochemical thrombus

formation was induced by irradiation with filtered green light in combination with intravenous

injection of the dye Rose Bengal. Formation of thrombus was continuously monitored for 30

min by utilizing a microcirculation dynamic viewing system and the thrombus size was

measured via image analysis. The results demonstrated that CA pretreatment delayed

thrombus-initiation time, and significantly reduced the area ratio of thrombus/venule. The

experiment explored the underlying mechanism and found that reactive oxygen species

production and von Willebrand factor secretion in human umbilical vein endothelial cells after

PS challenge was diminished by pretreatment with CA. The antioxidant potential of CA is

likely to be responsible for its beneficial effects on thrombosis through directly scavenging the

peroxides produced and /or indirectly depressing the release of von Willebrand factor in

vascular endothelial cell.

P3-3 Platelet Apoptosis in Uremic Patients Ming Li, Tong-Ling Ma, Guo-Yuan Lu, Kun Deng, Rong Yan and Ke-Sheng Dai*

The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Suzhou, China *Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: 0086-512-67781370; Fax: 0086-512-67781370 Introduction: Apoptosis, which serves for the regulation of cell lifespan and is long attributed exclusively to nucleated cells, has been well-documented in anucleate platelets. Diverse cell-external chemical and physical stimuli have been reported to induce transformation of resting platelets to apoptotic state. Uremia, a clinical syndrome associated with retention of various solutes that would normally be excreted by the kidneys, is frequently accompanied by bleeding tendency, and the mechanism remains unclear. The aim of the present study is to investigate whether platelet apoptosis occurs in uremia patients. Methods: Venous blood was drawn from 13 patients with end stage renal disease (ESRD) who exhibited uremia syndrome and 13 health controls. Platelet-rich plasma (PRP) was prepared and then was detected for apoptotic events including depolarization of mitochondrial inner membrane potential (ΔΨm), phosphatidylserine (PS) exposure, variations of apoptotic Bcl-2 family proteins, activation of caspases-3 by Flow cytometry or Western-blot. Furthermore, normal washed platelets were incubated with the poor- platelet plasma (PPP) from uremic patients, and then were detected for apoptotic events. The comparisons of the data were made using paired Student’s t-test. Results: Compared with controls, ΔΨm significantly depolarizated in platelets from uremic patients (P<0.05). Furthermore, Bax was up-regulated, Bcl-2 and Bcl-XL were down-regulated, and caspase-3 was activated in platelets from uremic patients. However, there was not significant difference in PS exposure and P-selectin expression between the platelets from uremic patients and health controls (P>0.05). Next, PPP from uremic patients was incubated with normal platelets, and the platelets presented ΔΨm depolarization, up-regulation of Bax, down-regulation of Bcl-2 and Bcl-XL, and caspase-3 activation. Conclusion: The data demonstrate that platelets are incurred apoptosis in uremia patients. The finding might suggest a novel pathogenic mechanism for bleeding tendency in uremic patients.

P3-4

Surface Expressions of Platelet Glycoprotein Ibα, GpⅡb/Ⅲa, and P-Selectin Are Elevated in Lung Cancer Patients Jian-Sheng Zhang, Li-Li Zhao, Jia-Nan Huang, Ping-Ping Zhang and Ke-Sheng Dai* The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Suzhou, China *Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: 0086-512-67781370; Fax: 0086-512-67781370 Objective: Thromboembolic events are common symptoms in patients with cancer, and the mechanisms are not totally understood. The platelet adhesion molecules play important roles in the process of thrombus formation. The aim of the current study is to investigate the surface expression of platelet glycoprotein (GP) Ibα, GPIIb/IIIa, and P-selectin in patients with lung cancer. Method: Platelets were isolated from patients with lung cancer and normal volunteers, and then the surface expressions of platelet GP Ibα, GPIIb/IIIa, and P-selectin were measured by Flow cytometry. The glycocalicin (GC) fragments in the peripheral blood of patients with lung cancer were tested by Western blot. The surface expression of GP Ibα on normal volunteers' platelets incubated with the platelet-poor plasma (PPP) of patients with lung cancer was measured by Flow cytometry and Western blot. Results: The surface expressions of platelet GP Ibα, GPIIb/IIIa, and P-selectin were obviously increased in patients with lung cancer compared with that of normal volunteers, and the GC fragments in PPP were also increased in patients with lung cancer. Incubation of control platelets with the PPP from cancer patients resulted in the elevation of the surface expression of GP Ibα in control platelets. Furthermore, the expressions of GP Ibα, GPIIb/IIIa, and P-selectin on platelets from patients with advanced stage of lung cancer (III-IV stage) were significantly higher than those with early stage (I-II stage) (P＜0.05). However, there were not significantly differences in the surface expressions of GP Ibα, GPIIb/IIIa, and P-selectin among the pathological types of lung cancer (squamous cell carcinoma, adenocarcinoma, small cell lung carcinoma, large cell lung carcinoma) (P＞0.05). Conclusion: The surface expressions of platelet GP Ibα, GPIIb/IIIa, and P-selectin were obviously elevated in patients with lung cancer, and the plasma from cancer patients is responsible for the elevations. These findings provide a new view to understand the pathogenic mechanisms of thromboembolic events in the patients with lung cancer.

P3-5

miR30c regulates plasminogen activator inhibitor-1 in platelet

Mao Luo, Qin Wan, Fei Liu, Rong Li, Ji-Yi Xia, Mei-Ping Ren, Ni Chen, Jian-Bo Wu*

Drug Discovery Research Center, Luzhou Medical College, Luzhou, Sichuan, China

Objective PAI-1 mRNA and protein have been detected in human platelets. Recently some

miRNAs have been found in human platelets, which are involved in the regulation of genes and

the protein synthesis. However, little is known about the physiological roles of individual

miRNAs in platelets. In this study, we investigated whether miR30c can regulate

platelet-derived plasminogen activator inhibitor-1(PAI-1).

Methods and Results Expression of miR-30c, PAI-1, miR-21 and its targeted gene TIMP1

were found in healthy human leukocyte-depleted platelets (LDPs) by real time PCR. In

luciferase reporter gene assay, miR-30c targets the 3' untranslated region (3' UTR) of PAI-1

mRNA through a miR-30c binding site. Transfection of miR-30c mimic into MEG-01, a

megakaryoblastic cell line, significantly reduced PAI-1 protein level compared with negative

control. Inhibition of miR-30c by transfecting miR-30c inhibitor significantly increased PAI-1

protein level. Furthermore, miR-21 expression was significantly down-regulated after

transfecting with miR-30c mimic in PAI-1-/- mice LDPs, conversely, the expression of its target

gene TIMP1 was significantly up-regulated after transfecting with miR-30c mimic in PAI-1-/-

mice LDPs.

Conclusion These results provide a novel regulatory mechanism of miR30c- regulated PAI-1

protein through its influence on the downstream miR21 and its target gene TIMP1 expression

in platelet, suggesting that miR-30c might be a potential new strategy for anti-thrombosis.

P3-6 Aspirin Induces Platelet Apoptosis Li-Li Zhao1,2, Wei-Lin Zhang1, Meng-Xing Chen2, Jian-Sheng Zhang2, Ming-Yi Zhang2, Chang-Geng Ruan, MD, PhD2 and Ke-Sheng Dai2*

1School of Biological Science and Medical Engineering, Beijing University of Aeronautics and Astronautics, Beijing, China; 2Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, Jiangsu Institute of Hematology, Suzhou, China *Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: 0086-512-67781370; Fax: 0086-512-67781370 Aspirin is widely used in the treatment of a number of clinical conditions, such as fever, pain, strokes, and heart attacks. Although aspirin is being thought of a relatively “safety” medicine, it also has some side effects, particularly the risk of bleeding which may be severe and lead to death. The mechanisms, however, are not totally understood. The effectiveness of aspirin has been attributed to its ability to inhibit prostaglandin production by inhibiting the cyclooxygenase (COX) enzyme. Interestingly, it has been reported recently that aspirin induces apoptosis in many cell types. Platelet apoptosis induced by either physical or chemical compounds or platelet storage occurs widely. We have reported that the glycoprotein Ibα-von Willebrand factor interaction, hyperthermia, and calmodulin antagonists induce platelet apoptosis, suggesting that platelet apoptosis might play important roles in controlling the number and function of circulated platelet under physiological conditions, or in the development of platelet-related hemorrhagic diseases. Thus, the aim of the current study is to explore whether aspirin induces platelet apoptosis. Washed platelets (3×108 /ml) were incubated with various concentrations of aspirin (2.5 mM, 5 mM, 10 mM, 20 mM) at 37 °C for 2 hours. The effect of aspirin on platelet mitochondrial membrane potential (ΔΨm) depolarization and phosphatidylserine (PS) exposure were analyzed by cell-permeable lipophilic cationic dye tetramethylrhodamine ethyl ester (TMRE) and annexin V binding, respectively. The data show that ΔΨms depolarization and PS exposures were dose-dependently induced by aspirin in platelets. In order to further confirm that aspirin incurs platelet apoptosis, caspase-3 activation was measured in platelets incubated with aspirin by detecting 32 kD pro-caspase-3 and 17 kD cleaved caspase-3 fragment, and the result shows that aspirin induced caspase-3 activation. Furthermore, platelet shrinkage appeared in platelets incubated with aspirin (20 mM). Caspase inhibitor z-VAD-fmk inhibited apoptotic platelet shrinkage, and blocked ΔΨm depolarization, but had no effect on PS exposure, suggesting that caspase-3 acts upstream of ΔΨm depolarization, and plays a key role in aspirin-induced platelet apoptosis. Whereas, protein kinase C (PKC), protein kinase B (PKB), p38 mitogen-activated protein kinases (MAPK), protein 53 (p53), reactive oxygen species (ROS), and mitogen-activated protein kinase kinase (MAPKK) were not involved in aspirin-induced platelet apoptosis. In addition, platelets incubated with another COX inhibitor indomethacin did not incur ΔΨm depolarazation and PS exposure in platelets. Taken together, the data indicate that aspirin induces platelet apoptosis via caspase-3 activation, and independent of the COX signaling.

Poster 4 Lymph Chair: Hui-Ling Huang Neuro-biochemical Lab

Neurological Institute of Tianjin Tianjin Neurological Hospital

13:20-13:30 P4-1 Nitric oxide is involved in the regulating of lymphatic reactivity following hemorrhagic shock Li-Min Zhang Institute of Microcirculation, Hebei North University 13:30-13:40 P4-2 Post-hemorrhagic shock mesenteric lymph is involved in cardiac dysfunction following hemorrhagic shock Hui-Bo Du Institute of Microcirculation, Hebei North University 13:40-13:50 P4-3 Role of hydrogen sulfide on post-hemorrhagic shock mesenteric lymph drainage alleviating multiple organ injury in rats Bo Han Institute of Microcirculation, Hebei North University 13:50-14:00 P4-4 Role of myosin light chain kinase on post-hemorrhagic shock mesenteric lymph blockade ameliorating the vascular reactivity and calcium sensitivity in rats following hemorrhagic shock Yu-Ping Zhang Institute of Microcirculation, Hebei North University

14:00-14:10 P4-5 Kudiezi injection adjust PKC δ/MARCKS signal pathway and protect vascular endothelial cells after hypoxia injury Fu-Qin Chen Department of Traditional Chinese Medicine, Peking University People’s Hospital

P4-1

Nitric oxide is involved in the regulating of lymphatic reactivity following hemorrhagic shock

Li-Min Zhang, Yu-Ping Zhang, Zi-Gang Zhao, Chun-Yu Niu* Institute of Microcirculation, Hebei North University, Zhangjiakou, China Objective: To investigate the effects of Nitric oxide (NO) on the shock-induced biphasic change in lymphatic reactivity and the related mechanism. Methods: The contents of NO, cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), phospho-PKA (p-PKA), and p-PKG were detected in thoracic ducts tissue obtained from hemorrhagic shocked rats. The roles of cAMP-PKA- KATP and cGMP-PKG- KATP in NO regulation of lymphatic reactivity to gradient substance P (SP) were observed in isolated lymphatics from control, 0.5 h- and 2 h-shocked rats with an isolated lymphatic perfusion system at a transmural pressure of 3 cmH2O. Results: The concentrations of NO, cAMP, cGMP, p-PKA and p-PKG were increased gradually along with the development of shock. The differences between the pre- and post- administration values of contractile frequency (CF), tonic index (TI), and fractional pump flow (FPF) of shock 0.5 h-lymphatics were increased, and in shock 2 h group were decreased significantly compared with the control group at certain concentration conditions of SP, respectively. The NO donor (L-Arg), 8-Br-cAMP (PKA donor) decreased the differences of CF, TI and FPF of shock 0.5 h-lymphatics, and the PKA inhibitor (H89) and KATP inhibitor (glibenclamide) restrained the effects of L-Arg on these indices, glibenclamide abolished the effects of 8-Br-cAMP at certain concentration of SP. NOS antagonist (L-NAME) and PKG inhibitor (KT-5823) increased the differences of CF, TI and FPF, soluble guanylate cyclase inhibitor (ODQ) increased the differences of CF, TI of shock 2 h-lymphatics, respectively; KATP opener (pinacidil) decreased the effects which were elevated by L-NAME, ODQ and KT-5823. Conclusions: NO regulation of the lymphatic reactivity after shock is closely related to cAMP-PKA-KATP and cGMP-PKG-KATP pathways. These findings have potential significance for the treatment of lymphatic hypo-reactivity. Acknowledge: This work was supported by the National Natural Science Foundation of China (30770845), the Natural Science Foundation of Hebei Province (C2008000503) and the Key Research Program of Education Department in Hebei Province (ZH2007101 and ZD20100201).

P4-2 Post-hemorrhagic shock mesenteric lymph is involved in cardiac dysfunction following hemorrhagic shock Hui-Bo Du, Si-Hai Wang, Li-Feng Chen, Li-Min Zhang, Zi-Gang Zhao, Chun-Yu Niu*

Institute of Microcirculation, Hebei North University, Zhangjiakou, China Objective: The present study was designed to evaluate whether post-hemorrhagic shock mesenteric lymph (PHSML) involved in cardiac dysfunction following hemorrhagic shock. Methods: The hemorrhagic shock model (40±2 mmHg, 3 h) was established in the shock and shock+drainage groups; and the PHSML was drained from hypotension 1-3 h in shock+drainage rats. Firstly, the effect of PHSML drainage on ventricular pressure in vivo was observed. Secondly, the isolated hearts were obtained from the rats of sham, shock and shock+drainage groups, and the cardiac function was examined with Langendorff heart perfusion system in vitro. Finally, the isolated hearts were obtained from normal rats and perfused with PHSML or K-H liquid, respectively; and the changes of cardiac function were observed at different time points. Results: In vivo, in the shock and shock+drainage groups, the left ventricular systolic pressure (LVSP) was increased, the left ventricular end diastolic pressure (LVEDP), heart rate (HR) and ±dP/dtmax were decreased compared with the sham group, respectively; but the HR and ±dP/dtmax in the shock+drainage group were increased than that of the shock group. In vitro, the LVSP and ±dP/dtmax in the shock and shock+drainage groups were lower than that of the sham group; otherwise, these indices in the shock+drainage group were higher than the shock group. In addition, after isolated hearts obtained from normal rats perfusing with PSML, these cardiac function indices were gradual decline along with the extension of time, such as HR, LVSP, ±dP/dtmax, etc. Conclusions: PHSML is an important contributor to cardiac dysfunction following hemorrhagic shock, suggesting that blockade of PHSML may be applied as an option for preventing cardiac remodeling after hemorrhagic shock. Acknowledge: This work was supported by the Talent Cultivation Project Funds of Hebei Province.

P4-3 Role of hydrogen sulfide on post-hemorrhagic shock mesenteric lymph drainage alleviating multiple organ injury in rats Bo Han, Li-Min Zhang, Li-Qiang Xing, Shu-Guang Li, Zi-Gang Zhao, Chun-Yu Niu*

Institute of Microcirculation, Hebei North University, Zhangjiakou, China Objective: To investigate the role of hydrogen sulfide (H2S) on post-hemorrhagic shock mesenteric lymph (PHSML) drainage alleviating multiple organ injury in rats, by administering D,L-propargylglycine (PPG) and sodium hydrosulfide hydrate (NaHS) to rats exhibiting PHSML drainage. Methods: Hemorrhagic shock model was established in the shock, shock+drainage, shock+drainage+PPG (45 mg/kg, 0.5 h pre-hemorrhage) and shock+drainage+NaHS (28 µmol/kg, 0.5 h pre-hemorrhage) groups; fluid resuscitation was performed after 1 h hypotension, and PHMSL was drained in the last three groups 3 h upon resuscitation. Organic function indices and histomorphology were assessed, along with H2S, cystathionine-γ-lyase (CSE), Toll-like receptor 4 (TLR4), interleukin-10 (IL-10), IL-12, and tumor necrosis factor α (TNFα) levels in lung, kidney, liver and myocardium. Results: Hemorrhagic shock induced multiple organ injury, including increased aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bile acid (TBA), urea, creatinine (Cre), lactic acid dehydrogenase-1 (LDH-1), creatine phosphokinase isoenzyme (CK-MB) and H2S in plasma, higher numbers of cells in bronchoalveolar lavage fluid (BALF) and lung permeability index (LPI), and enhanced H2S, CSE, TLR4, IL-10, IL-12, and TNFα in lung, kidney, liver and myocardium. PHSML drainage significantly reduced numbers of cells in BALF and LPI, AST, ALT, TBA, urea, Cre, H2S in plasma, H2S, CSE and TNFα in renal tissue, H2S, CSE, IL-10, IL-12 and TNFα in hepatic tissue, H2S, CSE, IL-12 and TNFα in pulmonary tissue, H2S and TNFα in myocardial tissue. Meanwhile, PPG decreased numbers of cells in BALF and LPI, AST, ALT, TBA, Cre, H2S in plasma, H2S, IL-10 and TNFα in renal tissue, H2S and TNFα in hepatic tissue, H2S, IL-10, IL-12 and TNFα in pulmonary tissue, IL-10 and TNFα in myocardial tissue. However, these beneficial effects of PSHML drainage were significantly reversed by NaHS administration. Conclusions: PHSML drainage alleviated multiple organ injury following hemorrhagic shock, resulting in reduced H2S and alleviated H2S-mediated inflammation. Acknowledge: This work was supported by the Talent Cultivation Project Funds of Hebei Province.

P4-4 Role of myosin light chain kinase on post-hemorrhagic shock mesenteric lymph blockade ameliorating the vascular reactivity and calcium sensitivity in rats following hemorrhagic shock Yu-Ping Zhang, Li-Min Zhang, Zi-Gang Zhao*, Chun-Yu Niu* Institute of Microcirculation, Hebei North University, Zhangjiakou, China

Objective: To explore whether myosin light chain kinase (MLCK) involves in the post-hemorrhagic shock mesenteric lymph (PHSML)-mediated vascular hyporeactivity and calcium desensitization in rats. Methods: Rats were divided into the sham, shock, shock+ligation and shock+drainage groups, and the hemorrhagic shock model (40±2 mmHg, 3 h) was established in the shock, shock+ligation and shock+drainage groups, the mesenteric lymph duct (MLD) was ligated or PHSML was drained from immediate hypotension for 3 h in the shock+ligation or shock+drainage rats, respectively. Superior mesenteric artery (SMA) tissue was obtained from rats for determining the phospho-MLCK (p-MLCK) and MLCK levels. And the SMA rings were prepared for observing the vascular reactivity and calcium sensitivity to norepinephrine (NE) and Ca2+ by an isolated organ perfusion system. Results: The levels of p-MLCK and MLCK were significantly decreased in the shock group, and that of the shock+ligation and shock+drainage group were observably increased compared with the shock group. Moreover, the hemorrhagic shock resulted in a significant decrease in the SMA reactivity and calcium sensitivity at multiple concentrations and Emax, which was enhanced by MLD ligation and PHSML drainage. Meanwhile, this beneficial effect of the MLD ligation and PHSML drainage were further ameliorated by the substance P (SP, 1 nmol/L), an agonist of MLCK, were abolished by the ML-7 (0.1 nmol/L), an inhibitor of MLCK. Conclusions: MLCK is an important contributor to PHSML drainage or MLD ligation enhancing the vascular reactivity and calcium sensitivity in hemorrhagic shock rats, suggesting that blockade of PHSML may be applied as an option for preventing vascular hypo-reactivity and calcium desensitization after hemorrhagic shock. Acknowledge: This work was supported by the National Natural Science Foundation of China (30971203) and National Natural Science Foundation of Hebei Province (C2012405020).

P4-5 Kudiezi injection adjust PKC δ/MARCKS signal pathway and protect vascular endothelial cells after hypoxia injury Fu-Qin Chen and Jia-Lan Yu1, 2, Jia-Yi Yang2, Wen Bai2*

1 Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, China

2 Department of Traditional Chinese Medicine, Peking University People’s Hospital, Beijing,

China

Objective: The present study was designed to evaluate whether Kudiezi injection was beneficial for vascular endothelial cells after hypoxia injury and prove the role of PKC

δ/MARCKS signal pathway on the vascular endothelial cells after hypoxia injury;

Methods: Induce by CoCl2 establish human umbilical vein endothelial cells (HUVEC) hypoxia injury model, and apply Kudiezi injection and PKCδ specific inhibitor Rottlerin to

intervene. To observe the morphology of HUVEC after hypoxia injury. Determined by MTT

colorimetry to detect the vitality of HUVEC after hypoxia injury. The distribution of p-PKCδ,

p-MARCKS protein were determined by immunocytochemical method. The expression of

p-PKCδ, p-MARCKS protein were determined by western blotting.The expression of p-PKCδ,

p-MARCKS mRNA were determined by Quantitive Real time PCR.

Results: Kudiezi injection and Rottlerin can reduce the HUVEC morphology damage after hypoxia. Kudiezi injection and Rottlerin can significantly enhance the vitality of HUVEC after

hypoxia injury (P<0.05 or P<0.01). The p-PKCδ, p-MARCKS protein were mainly distributed

in the cell membrane. The expression of p-PKCδ, p-MARCKS protein increase anomalously in

HUVEC after hypoxia injury, Kudiezi injection and Rottlerin can reduce the expression

（P<0.05）. The expression of p-PKCδ, p-MARCKS mRNA increase anomalously in

HUVEC after hypoxia injury, Kudiezi injection and Rottlerin can reduce the expression

（P<0.05）, there was no significant difference between two groups.

Conclusions: Kudiezi injection can protect vascular endothelial cells after hypoxia injury, the mechanism related to the PKC δ/MARCKS signal path.

Poster 5 Ischemia-Reperfusion injury and microcirculation

Chair: Osamu Furukawa


13:20-13:30 P5-1 Inhibition of NADPH oxidase mediates protective effect of cardiotonic pills® against rat heart ischemia/reperfusion injury Xiao-Yuan Yang Tasly Microcirculation Research Center, Peking University Health Science Center 13:30-13:40 P5-2 Protective effects of Notoginsenoside R1 on intestinal ischemia/reperfusion injury in rats Chong Li Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University 13:40-13:50 P5-3 Inhibition of PAI-1 rescues the defect in ischemia-induced neovasculature in type II diabetic mice Rong Li

Drug Discovery Research Center, Luzhou Medical College 13:50-14:00 P5-4 Mechanism underlying trichosanthes pericarpium injection on acute myocardial ischemia protection

Qi-Tao Zhao School of Basic Medicine, Shandong University of Traditional Chinese Medicine 14:00-14:10 P5-5 Oregonin from the bark of Alnus japonica improves ischemia-reperfusion induced microcirculatory disturbance in rat mesentery Nguyen Huu Tung Faculty of Pharmaceutical Sciences, Nagasaki International University 14:10-14:20 P5-6 Advanced glycation end products induce endoplasmic reticulum stress through reactive oxygen species in endothelial cells Li-Li Wu Department of Pathophysiology, Key Lab for Shock and Microcirculation Research, Southern Medical University

P5-1

Inhibition of NADPH oxidase mediates protective effect of cardiotonic

pills® against rat heart ischemia/reperfusion injury

Xiao-Yuan Yang1,2, Na Zhao1, Yu-Ying Liu1, Bai-He Hu1, Kai Sun1, Xin Xue5, Xin Chang1,

Xiao-Hong Wei1, Jing-Yu Fan1, Jing-Yan Han1,2,3,4

1 Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing,

China 2 Department of Integration of Traditional Chinese and Western Medicine, School of Basic

Medical Sciences, Peking University, Beijing, China 3 Key Laboratory of Microcirculation, State Administration of Traditional Chinese Medicine of

the People's Republic of China 4 Key Laboratory of Stasis and Phlegm, State Administration of Traditional Chinese Medicine

of the People's Republic of China 5Institute of Basic Theory of TCM, China Academy of TCM

Cardiotonic pills® (CP) is a compound Chinese medicine currently used in China for treatment

of ischemic angina pectoris. Our previous results indicated that a single dosing of CP

pretreatment at 0.8 g/kg attenuates ischemia/reperfusion (I/R)-induced myocardial injury and

cardiac microcirculatory disturbance. The present study aimed to investigate the effect of CP at

low dosage in a multiple dosing manner, and to uncover the mechanism of antioxidative

activity of CP. Male Sprague-Dawley rats were subjected to left anterior descending artery

occlusion for 30 min followed by 60 min reperfusion. CP was administrated daily by gavage

for six days at 0.1, 0.4, and 0.8 g/kg/day before I/R. Results showed that multiple dosing of CP

at three doses significantly reduced I/R induced myocardial injury, microcirculatory

disturbance and oxidative stress. CP dramatically inhibited I/R-induced nicotinamide adenosine

dinucleotide phosphate (NADPH) oxidase subunit gp91phox expression, and p67phox and p47phox

translocation from cytosol to cell membrane. Translocation of cytosolic subunits to membrane

are required for the activation of NADPH oxidase. These data suggested that multiple dosing

of CP at doses ranging from 0.1 to 0.8 g/kg/day reduced I/R-induced rat myocardial injury and

microcirculatory disturbance, which was mediated by inhibition of NADPH oxidase activation.

P5-2

Protective effects of Notoginsenoside R1 on intestinal ischemia/reperfusion injury in rats Chong Li 1,2,3, Quan Li 2,3,4, Yu-Ying Liu 2,3,4, Ming-Xia Wang 2,3,4, Chun-Shui Pan 2,3,4, Li Yan 2,3,4, Yuan-Yuan Chen 1,2,3, Jing-Yu Fan 2,3,4 and Jing-Yan Han 1,2,3,4*

1Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China 2Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China 3Key Laboratory of Microcirculation, State Administration of Traditional Chinese Medicine, China 4Key Laboratory of Stasis and Phlegm of State Administration of Traditional Chinese Medicine, Beijing, China Intestinal ischemia and reperfusion (I/R) is a clinical problem occurred for diverse causes with high mortality. Prophylaxis and treatment of intestinal I/R remains a challenge for clinician. The present study was to explore the role of Notoginsenoside R1 (R1), a major component form Panax. Notoginseng, in management of intestinal I/R injury. Intestinal I/R was induced in male Sprague-Dawley rats by clamping the superior mesenteric artery for 90 min followed by reperfusion for 60 min or 3 days. R1 (10 mg/kg/h) was administered either 20 min before ischemia or 20 min after reperfusion. Intestinal microcirculation was evaluated by intravital microscopy over 60 min reperfusion. Sixty min or 3 days after reperfusion, rats were killed for histological examination of the jejunum tissue and immunohistochemical localization of myeloperoxidase and CD68. ATP, ADP and AMP content in jejunum tissue was assessed by ELISA. Activation of NF-κB, expression of ATP5D and tight junction proteins were determined by Western blotting. The results demonstrated that R1 is capable of attenuating intestinal I/R-induced microvascular hyperpermeability, inflammatory cytokine production, NF-κB activation and loss of tight junction proteins, as well as improving energy metabolism during I/R. The results of the present study suggest R1 as an option in protecting against intestinal I/R injury.

P5-3

Inhibition of PAI-1 rescues the defect in ischemia-induced neovasculature in type II diabetic mice Rong Li, Xiao Zhang, Mei-Ping Ren, Mao Luo, Kai Yan, Jian-Bo Wu*

Drug Discovery Research Center, Luzhou Medical College, Luzhou, Sichuan, China

Objective- Type II diabetes mellitus (DM) leads to the development of microvascular diseases

and is associated with impaired angiogenesis. Elevated plasma levels of plasminogen

activator inhibitor-1 (PAI-1) are associated with DM. However, the role of PAI-1 in the

development of DM-associated vascular disease is poorly defined.

Methods and Results- Hind-limb ischemia was generated by femoral artery occlusion in

diet-induced diabetic mice and age-matched normal chow (NC)-fed mice. Plasma was

analyzed for glucose and PAI-1. PAI-039, a specific, small molecule pharmacological

inhibitor of PAI-1, was orally administered to mice (1 mg/kg, twice daily for 14 days) to

determine if PAI-1 attenuates neovasculature formation in DM mice. PAI-1 activity and antigen

were significantly increased within 2 weeks of DM onset and remained elevated throughout the

experimental period. PAI-039 normalizes elevated plasma PAI-1 activity in the DM group.

Color Laser Doppler showed that the blood flow perfusion in ischemic hind-limb was

significantly decreased in the DM nontreated group compared to NC nontreated group at 5, 7,

and 14 days (n = 6/group, P< 0.05). Decreased blood flow in the DM group correlated to

decreased density of CD31-positive vessels in ischemic muscle compared with the NC tissue.

Pharmacologic inhibition of PAI-1 with PAI-039 rescued the ischemia-mediated impairments

in blood flow perfusion and vessel density in ischemic muscles in DM group compared to

vehicle-DM group (n = 6/group, P < 0.05).

Conclusions- Taken together, these findings illustrate that the pharmacologic inhibition of

elevated PAI-1 rescues the impairments in ischemia-mediated flow perfusion and

neovasculature observed in DM, and may have efficacy as a therapeutic strategy to prevent

diabetic microangiopathy.

P5-4

Mechanism underlying trichosanthes pericarpium injection on acute myocardial ischemia protection Qi-Tao Zhao School of Basic Medicine, Shandong University of Traditional Chinese Medicine, Jinan, 250355, China. Objective: Trichosanthes pericarpium injection is widely used to cure coronary heart disease, myocardial ischemia patients in China. In previous studies, we find that t. pericarpium injection could significantly reduce the myocardial infarction rate and the serum activity of creatine-kinase (CK) et al in acute myocardial ischemia model rats. This study was designed to find out the mechanism underlying this pharmacodynamical protective effect. Methods: To make acute myocardial ischemia animal model, 1 hour after the last administration, the left anterior descending coronary branch were ligated in male wistar rats that have received abdominal cavity administration for 7 days. Rats in different groups were given trichosanthes pericarpium injection, the radix salviae miltiorrhizae injection (the positive control drug), and 0.9 % sodium chloride solution respectively. 6 h after of operation, the serum content of endothelin (ET), nitric oxide (NO), endothelial nitric oxide synthase (eNOS), malondialdehyde (MDA), superoxide dismutase (SOD), 6-ketone-PGF1a, and thromboxane (TXB2) in these rats were measured. Results: Compared with that in the model group rats, the myocardial infarction rate of rats in the trichosanthes group decreased evidently (P<0.05), the serum ET, MDA, TXB2 content of those rats was markedly reduced (P<0.05 or P<0.01), but the serum level of NO, 6-ketone-PGF1a, eNOS, and the activity of SOD in myocardial tissue increased observably (P<0.05 or P<0.01). Conclusion: These results suggest that, in the acute myocardial ischemic rats, trichosanthes pericarpium injection could obviously activate SOD and eNOS, promote the production of NO and 6-ketone-PGF1a, inhibit ET-1 and TXB2 synthesis, decrease the content of MDA in myocardial tissue. These data provided substantial evidence that, trichosanthes pericarpium injection may protect myocardium from ischemic injury by improving vasodilation so as to increase coronary blood flow, reducing oxidative stress, and inhibiting thrombosis. Key words: myocardial ischemia, trichosanthes pericarpium injection, endothelin, superoxide dismutase, thromboxane

P5-5

Oregonin from the bark of Alnus japonica improves ischemia-reperfusion induced microcirculatory disturbance in rat mesentery

Nguyen Huu Tung1, Kai Sun2, Jing-Yu Fan2, Jing-Yan Han2,3* and Yukihiro Shoyama1*

1Faculty of Pharmaceutical Sciences, Nagasaki International University, 2825-7 Huis Ten

Bosch, Sasebo, Nagasaki 859-3298, Japan. 2Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing,

100191, China 3Department of Integration of Chinese and Western Medicine, School of Basic Medical

Sciences, Peking University, Beijing 100191, China

Ischemia-reperfusion (I/R) injury occurs in a wide range of situations, including trauma,

vascular reflow after contraction, percutaneous transluminal coronary angioplasty,

thrombolysis treatment, organ transplantation, and hypovolemic shock with resuscitation.

I/R-induced microcirculatory disturbance with diverse manifestations is considered to be the

pathological basis of I/R injury. This study aimed to investigate, for the first time, potential

effects of oregonin (ORG), a major diarylheptanoid from the bark of Alnus japonica, on

I/R-induced rat mesenteric microcirculatory injury. ORG (5 mg.kg-1.h-1) was continuously

infused starting from 10 min before ischemia. The venule diameter, number of adherent

leukocytes, and mast cell degranulation were determined using an intravital microscope. The

results showed that the ORG treatment exerted an ameliorating effect on I/R-induced disorders,

characterized by decreasing the number of adherent leukocytes, attenuating the I/R-induced

dihydrorhodamine 123 fluorescence intensity on the venular walls, and reducing mast cell

degranulation, respectively.

P5-6

Advanced Glycation End Products Induce Endoplasmic Reticulum Stress through Reactive Oxygen Species in Endothelial Cells

Li-Li Wu, Da Wang, Li-Qun Wang, Bo Chen, Qiang Li, Qiao-Bing Huang

Department of Pathophysiology, Key Lab for Shock and Microcirculation Research, Southern Medical University, Guangzhou, 510515, P. R. China

Objective：Both advanced glycation end products (AGEs) and endoplasmic reticulum (ER) stress play important roles in the development of various diseases. The concomitant presence of advanced glycation in the same conditions with ER stress suggests their crosstalk in the progression of diseases associated with hypoxic and oxidative stress. This study aimed to clarify whether AGEs can induce ER stress in human umbilical venous endothelial cells (HUVECs). Methods: AGEs-induced ER stress was observed by the expression and activation of ER stress marker proteins GRP78, IRE1α and JNK using immunoblotting. The consequence of AGE-induced ER stress was revealed by the translocation of NF-κB to nucleus using immunoblotting and immunocytochemistry. The mechanism of AGE-induced ER stress was also explored by interfering the effects of reactive oxygen species (ROS) using siRNA and antioxidant. Results: The results demonstrated an enhanced expression of GRP78 in time and dose-dependent pattern in HUVECs exposed to AGEs. The application of AGEs also triggered the increases of phosphorylation of IRE1α and JNK in HUVECs. NF-κB p65 was observed to translocate from cytoplasmic region into the nucleus area in HUVECs treated with AGEs. Usages of Nox4 siRNA and antioxidant reduced glutathione (GSH) attenuated the AGE-induced expression of GRP78. The AGE-evoked activations of IRE1α and JNK were inhibited by Nox4 siRNA and GSH either. Nox4 and GSH also abolished the nuclear translocation of NF-κB in HUVEVs. Conclusion: This study confirmed AGEs induced ER stress in HUVECs by focusing on the ER stress-enhanced inflammatory response through the activation of JNK and NF-κB. This present report also further revealed the involvement ROS in mechanism of AGE-evoked ER stress.

Poster 6 Shock and inflammation

Chair: Chun-Yu Niu

Institute of Microcirculation, Hebei North University

13:20-13:30 P6-1 The effects of several vasoactive agent on intestines mesentery microcirculation in haemorrhagic shock Shang-Bin Yu Center Laboratory of Functional Science, Tongji Medical College, Huazhong University of Science and Technolgy 13:30-13:40 P6-2 Berberine inhibits Chlamydia pneumoniae infection-induced VSMC migration through downregulating expression of MMP3 and MMP9 via PI3K Jun-Yan Wei Department of Pathophysiology, School of Basic Medical Sciences, Tianjin Medical University 13:40-13:50 P6-3 Determination of sex hormone in the chronic myeloid leukemia patients treated with TKI Xiao-Hui Chang The 210th Hospital of PLA, TCM Hematology Center 13:50-14:00 P6-4 Epidemical survey of PLGC of phlegm-accumulation stasis and association with the polymorphism of TLR4 Gene Mu-Xin Wei

Institute of Integreted Medicine, Nanjing Medical University 14:00-14:10 P6-5 Huang Qi Jian Zhong Pellet® repairs colonic mucosal injury induced by 2,4,6-trinitrobenzene sulfonic acid in rats via mechanisms involving improvement of energy metabolism Duan-Yong Liu Jiangxi University of Traditional Chinese Medicine 14:10-14:20 P6-6 Protective effects of paeonol on lipopolysaccharide-induced mesenteric microcirculatory disturbance in rats Lei Dong

Department of Critical Care Medicine, Beijing Friendship Hospital, Capital Medical University

P6-1

The effects of several vasoactive agent on intestines mesentery microcirculation in haemorrhagic shock Shang-Bin Yu, Ying Yang, Hui-Yun Zhu Center Laboratory of Functional Science, Tongji Medical College, Huazhong University of Science and Technolgy. Wuhan 430030

AIM: To investigate the effect of three vasoactive drugs on mesenteric microcirculation during

hemorrhagic shock.

METHODS: Male Sprague-Dawley rats were divided into 6 groups which were control group

(group A), hemorrhagic shock group (group B), hemorrhagic shock group with normal saline

treatment (group C), hemorrhagic shock group with dopamine treatment (group D),

hemorrhagic shock group with noradrenalin treatment (group E), hemorrhagic shock group

with anisodamine treatment (group F). We employed bloodletting to decrease the Bp to the

level of 60mmHg to establish the animal model of hemorrhagic shock, and recorded

parameters to evaluate the effect of different types of drugs on hemorrhagic shock.

RESULTS: Hemorrhagic shock decreased the Bp, calibres of both inlet and oulet of the

mesenteric microcirculation, and the velocity of blood flow. Treatment of normal saline during

hemorrhagic shock could only increase Bp back to the normal level without improving the

mesenteric microcirculation. Dopamine treatment deteriorate the condition by attenuating Bp

and blood velocity, meanwhile expand the microcirculation (P＜0.05). Compare with group A,

noradrenalin treatment could partly reverse the impairment of Bp and mesenteric

microcirculation during hemorrhagic shock (P ＜ 0.05-0.01). Interestingly, only could

anisodamine rescue the parameters to the normal level.

CONCLUSION: Anisodamine with transfusion treatment could improve mesenteric

microcirculation during hemorrhagic shock. These data suggest that anisodamine could serve

as a potential therapeutic agent for preventing shock pathological changes via modulating

mesenteric microcirculation.

Keyword: Vasoactive agent, Haemorrhagic shock, Microcirculation, Anisodamine, Dopamine,

Noradrenalin

P6-2 Berberine inhibits Chlamydia pneumoniae infection-induced VSMC migration through downregulating expression of MMP3 and MMP9 via PI3K Jun-Yan Wei, Li-Jun Zhang, Bei-Bei Wang, Jun-Xia Zhang, Hai-Wei Wang, Li-Jun Zhang*

Department of Pathophysiology, School of Basic Medical Sciences, Tianjin Medical University Objective: To investigate the roles of matrix metalloproteinases (MMPs) in vascular smooth muscle cell (VSMC) migration induced by Chlamydia pneufmoniae (C.pn) infection, to observe the antagonistic effects of berberine on C.pn infection-induced VSMC migration, and to explore the related molecular mechanisms. Methods: A Matrigel invasion assay were performed to explore the roles of MMPs in VSMC migration induced by C.pn infection, and to observe the effects of berberine on the infection-induced VSMC migration. RT-PCR was used to detect the mRNA expressions of MMPs. After VSMCs were administrated with a PI3K specific inhibitor LY294002 or berberine at the different concentrations, gelatin zymography and ELISA were performed to examine the changes in MMP9 enzymatic activity and the protein expression levels of MMP3 and MMP9. Western blot was used to observe the phosphorylated Akt (p-Akt) expression levels in the infected VSMCs after the treatment with berberine. Results: The results from the invasion assay showed that the cells migrated through Matrigel induced by C.pn infection, after pretreated with GM6001, decreased (P < 0.05). RT-PCR results showed that the mRNA expressions of MMP3 and MMP9 in the infected VSMCs began to increase at 4 h and 18 h post-infection, and reached a peak at 18 h and 36 h, respectively, and both remained elevated until 72 h (P < 0.05). The results of gelatin zymography showed that the MMP9 activity was positively correlated with the time of C.pn infection in the infection period (r = 0.9829, P < 0.05), and the protein expressions of MMP3 and MMP9 by ELISA respectively began to rise at 12 h and 24 h post-infection, both reached a peak 72 h (P < 0.05). When the infected VSMCs were pretreated with LY294002, the MMP9 activity and protein expressions of MMP3 and MMP9 induced by C.pn infection all decreased (P < 0.05). After the infected VSMCs were administrated with berberine at the indicated concentrations, the number of VSMCs migrated through Matrigel, the MMP9 activity, the protein expressions of MMP3 and MMP9 induced by C.pn infection all reduced gradually with the increase in berberine concentrations (P < 0.05). Western blot analysis showed that berberine significantly decreased the expressions of p-Akt induced by the infection (P < 0.05). Conclusions: C.pn infection promotes VSMC migration possibly through up-regulating the expression levels of MMP3 and MMP9 via PI3K, and berberine may suppress the infection-induced VSMC migration through downregulating the protein expressions of MMP3 and MMP9 via PI3K. This work was supported by the National Natural Science Foundation (No.30971225) *Corresponding author: Lijun Zhang. E-mail address: [email protected]

P6-3 Determination of sex hormone in the chronic myeloid leukemia patients treated with TKI

Xiao-Hui Chang, Ai-Hong Wei The 210th Hospital of PLA, TCM Hematology Center, Dalian 116021 Objective: To investigate the changes of serum sex hormone level and tongue microcirculation for long-term TKI therapy in chronic myeloid leukemia (CML) patients and to understand the effects of TKI on reproductive system. Methods: 56 cases of CML patients were treated with TKI for more than 3 years, including 24 males, 32 females, the median age was respectively males 47.5 (16-58), females 36.7 (21-52); 30 cases in each control group, who were healthy blood donors, the median age was respectively males 42.5 (26-28), females 32.4 (21-52); to detect the levels of sex hormones include: testosterone (T), progesterone (P), follicle stimulating hormone (FSH), Latinizing hormone (LH), estradiol (E2) and prolactin (PRL) in each group by the chemiluminescence immunoassay. Tongue tip organizations and microvascular structure were observed by using multi-site microscopic LAU type WX-6. Results: The sex hormone levels in male CML patients group and male control group were as follows: T 12.47±3.88/10.78±4.16 ug/l; P 2.37±0.97/ ng/ml; FSH 4.56±1.09/5.13±1.65 mIU/ml; LH 5.36±1.58/7.03±2.16 mIU/ml; E2 52.28±9.95/ 47.98±7.65 pg/ml; PRL 235.47±145.6/195.47±51.1 ng/ml; the sex hormone levels in female CML patients group and female control group were as follows: T 0.59±0.09/0.68±0.06 ug/l; P 1.41±0.39/ ng/ml; FSH 24.11±3.56/25.7±5.01 mIU/ml; LH 16.31±2.79/17.93±2.62 mIU/ml; E2 49.76±6.57/ 53.98±6.19 pg/ml; PRL252.35±127.64 /215.47±31.1 ng/ml; CML patients compared with normal controls after TKI treatment, appearing the following situation: mushroom nipple decreased significantly, spines desmosome connection structure clearly, cell arrangement structure messy, the epithelium cornification significantly increased, scattered basal cell, cell swelling, sizes, and gap base layer organization coarse, base layer under the capillary tube cavity expansion, endothelial cell swelling, capillary lumen, lumen leukemia cells increased. Conclusion: Long-term TKI therapy in the patients with CML had no obvious influence for hormone level, but there is a certain degree of tongue microcirculation dysfunction.

P6-4 Epidemical Survey of PLGC of Phlegm-Accumulation Stasis and Association with the Polymorphism of TLR4 Gene

Mu-Xin Wei1, 2, Yao-Fu Fan1, Yan-Min Wu2, Hao Liu1

1. Institute of Integreted Medicine, Nanjing Medical University 2. Department of Traditional Chinese Medicine, The First Affiliated Hospital with Nanjing Medical University Objective: To investigate the association between various risk factors and precancerous lesion of gastric cancer (PLGC) in patients from Jiangsu. This study aimed to assess Toll-like receptor 4 (TLR4) (rs4986790, rs4986791, rs10759932) polymorphisms at risk of precancerous lesion of gastric cancer (PLGC) of Phlegm-Accumulation Stasis in a Chinese Han population. Methods: 501 cases of PLGC of Phlegm-Accumulation Stasis and 523 cases of superficial gastritis were included. A comparative study of the relation between different risk factors and PGLC of Phlegm-Accumulation Stasis was performed. In this hospital-based case-control study, gene polymorphisms were analyzed in 333 patients (Intestinal metaplasia: n=193; Intraepithelial neoplasia: n=140) and 312 atopy-free controls of Chinese Han population. TLR4 (rs4986790, rs4986791, rs10759932) were performed using Taqman allelic discrimination assays. Result: 1. Statistical differences were noted in a series of indexes including Helicobacter pylori (HP) infection, family history of esophageal cancer (EC), gastric cancer (GC) and chronic atrophic gastritis (CAG), personal history of CAG, gastric polyps (GP) and gastric ulcer (GU), usage of non-steroids (e.g. aspirin), gastroesophageal reflux disease (GERD), drinking alcohol, eating food rich in nitroso compounds, irregular eating habits with no breakfast, ingestion of smoked meat, fried food and spicy food, anxiety and depression. 2. The risk factors associated with PLGC ranked in an order of personal history of CAG, GP, family history of GC, usage of non-steroids (e.g. aspirin), ingestion of spicy food frequently, personal infection with HP, family history of EC, drinking alcohol, anxiety, personal history of GU, GERD and family history of CAG. (3) The single nucleotide polymorphisms TLR4 rs4986790, rs4986791 was not associated with risk of chronic gastritis or gastric cancer and the homozygous genotypes TLR4 rs4986790 GG and TLR4 rs4986791 TT were absent in the studied population. In the single-locus analysis, the C allele of rs10759932 had a significantly reduced risk of IM and IN (adjusted odds ratio (OR) = 0.42; 95%CI = 0.29-0.62 and OR=0.62; 95%CI = 0.41-0.93, respectively), compared with the wild-type homozygote (TT). The frequency of the TLR4 rs10759932 TC tended to be lower in patients with IM and IN than in controls (OR= 0.37; 95%CI = 0.24-0.55 and OR=0.60; 95%CI = 0.39-0.93, respectively). The frequencies of TLR4 rs10759932 TC and T carrier were significantly lower in patients with Sydney’s slight IM and LGIN (P < 0.01 and P = 0.01, respectively), meanwhile the TC genotype showed a lower risk of moderate IM compared to healthy individuals (P=0.045). There were no significant differences between these groups in the distribution of age, sex and smoking status. But Helicobacter pylori infection, drinking, salt uptake were associated with a higher susceptibility to developing this neoplasm. Conclusion: 1. Personal history of CAG was most associated with PLGC in patients from eastern China, followed by personal history of GP and family history of GC. 2. Our data suggests that TLR4 rs10759932 TC and C carrier were associated with a lower risk of developing PLGC of Phlegm-Accumulation Stasis, and these genotypes may reduce the risk of IM and IN in a Chinese Han population.

P6-5 Huang Qi Jian Zhong Pellet® repairs colonic mucosal injury induced by 2,4,6-trinitrobenzene sulfonic acid in rats via mechanisms involving improvement of energy metabolism Duan-Yong Liu1.2, Chun-Shui Pan2, Yu-Ying Liu2, Xiao-Hong Wei2, Chang-Man Zhou2, Kai Sun2, Ke He2, Chong Li2, Li Yan2, Jing-Yu Fan2, Chuan-She Wang2.3.4.5, Hong-Ning Liu1* and Jing-Yan Han2.3.4.5*

1 Jiangxi University of Traditional Chinese Medicine, Nanchang, China. 2 Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China 3 Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China 4 Department of Anatomy, School of Basic Medical Sciences, Peking University, Beijing, China 5 Key Laboratory of Microcirculation, State Administration of Traditional Chinese Medicine, Beijing, China

Huang Qi Jian Zhong Pellet (HQJZ) is a famous Chinese medicine formula used to treat various gastrointestinal tract diseases. To explore the mechanism thereby HQJZ repairs the colonic mucosal injury, this study investigated the role of HQJZ in IBD and its underlying mechanism. Colonic mucosal injury was induced by 2,4,6-trinitrobenzene sulfonic acid in the Sprague-Dawley rats. In the HQJZ treatment group, HQJZ was administered (2 g/kg) for 14 days starting from 1 day after TNBS infusion. The effects of HQJZ were evaluated by macroscopic and microscopic examination, colonic microcirculation and blood flow by inverted intravital microscopy and a Laser-Doppler perfusion imager, respectively. Colonic mucosal injury occurred obviously 1 day after TNBS challenge, and did not recover distinctively till day 15, including an increase in macro- and microscopic score, colonic weight index, a decrease in colonic length, number of functional capillaries, and blood flow. TNBS-induced colonic microcirculatory disturbance included increased leukocyte rolling in, adhesion to and albumin leakage from colonic venules. Assessment by ELISA revealed an increment in TNF-α and IL-6, and a decrease in IL-10 in colonic tissue in response to TNBS. In an attempt to address the mechanism behind these injuries, occludin, RhoA and ROCK-I were observed to be decreased, while Rac-1, PAK-1 and phosphorylated myosin light chain increased by TNBS. Finally, ELISA and Western blot showed a reduction in ATP and ATP5D after TNBS challenge. Impressively, treatment with HQJZ for 14 days significantly attenuated all the alterations evoked by TNBS, promoting the recovery of colonic injury. The present study demonstrated HQJZ as a multitargeting management for colonic mucosal injury, which set in motion mechanisms involving improvement of energy metabolism.

P6-6

Protective Effects of Paeonol on Lipopolysaccharide -Induced Mesenteric Microcirculatory Disturbance in Rats

Lei Dong1, Ang Li1, Yu-Ying Liu2, Kai Sun2, Mei-Li Duan1 and Jing-Yan Han2,3

1 Department of Critical Care Medicine, Beijing Friendship Hospital, Capital Medical

University, Beijing, China 2 Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing,

China 3 Department of Integration of Chinese and Western Medicine, School of Basic Medical

Sciences, Peking University, Beijing, China

Aim: To investigate the effect of paeonol on lipopolysaccheride (LPS)-induced rat mesenteric

microcirculatory dysfunctions.

Methods: Male Wistar rats were randomly distributed into 5 groups (n=6 each): Sham group,

LPS group, paeonol group, paeonol +LPS group, LPS +paeonol group. Sepsis models were

conducted by continuous LPS infusion. Changes in mesenteric microcirculatory variables,

including diameter of venule, velocity of red blood cells in venule, leukocyte adhesion, free

radicals released from venule and albumin leakeage, were observed through an inverted

intravital microscope. Meanwhile, the expression of TLR4, NF-κB, ICAM-1, MPO, CD-18,

AP-1 and JNK was evaluated by western-blot.

Results: After infusion of LPS, the number of leukocytes adherent to venular wall, the

intensity of Dihydrorhodamine 123 (DHR) fluorescence in the venular walls, and albumin

leakage from venules were significantly increased, whereas the red blood cell velocity in

venule was decreased. All the manifestations were significantly reduced by pretreatment and

post-treatment with paeonol. Moreover, paeonol significantly attenuated the expression of

TLR4, NF-κB, ICAM-1, MPO, CD-18, AP-1 and JNK in rat mesentery.

Conclusion: The results demonstrated that paeonol could protect from and ameliorate the

microcirculation disturbance induced by LPS.

Poster 7 Brain injury and microcirculation Chair: Shi-Jun Wang

Shandong University of Traditional Chinese Medicine 13:20-13:30 P7-1 Long-term stimulation with electroacupuncture at DU20 and ST36 attenuates cerebral blood flow and rescues hippocampal neuron in spontaneously hypertensive rats Gui-Hua Tian

School of Acupuncture and Moxibustion, Beijing University of Chinese Medicine 13:30-13:40 P7-2 Adoptive regulatory T-cell therapy prevents cerebral vasospasm and reduces early brain injury following experimental SAH Li-Ping Zhang Department of Neurology and Key Lab oratory of Cerebral Microcirculation, Taishan Medical University 13:40-13:50 P7-3 Intravenous transplantation of In vitro expanded CD4+CD25+ regulatory T cells exerts protective effects and promotes the expression of Iba1, GFAP after cerebral Ischemia-Reperfusion in rats Yan Cheng Key Lab of Cerebral Microcirculation in Universities of Shandong (Taishan Medical University); Department of Neurology, Affiliated Hospital of Taishan Medical University 13:50-14:00 P7-4 Anti-inflammation treatment after subarachnoid hemorrhage: regulatory T-cells adoptive transfer Lei Zhang Key Lab of Cerebral Microcirculation in Universities of Shandong (Taishan Medical University); Department of Neurology, Affiliated Hospital of Taishan Medical University 14:0-14:10 P7-5 Blocking neurodegenerative damages of Alzheimer's disease by aromatic glycosides in multiple pathways Ying-Hong Wang Department of Combined Traditional Chinese Medicine with Western Medicine, School of Basic Medical Science, Peking University 14:10-14:20 P7-6 The involvement of programmed cell death 5 (PDCD5) in the regulation of apoptosis in cerebral ischemia/reperfusion injury Chun-Hua Chen Department of Anatomy and Embryology, Peking University Health Science Center

P7-1 Long-term stimulation with electroacupuncture at DU20 and ST36 attenuates cerebral blood flow and rescues hippocampal neuron in spontaneously hypertensive rats Gui-Hua Tian1,2,3, Kai Sun3,6, Ping Huang3,6, Chang-Man Zhou4, Hai-Jiang Yao1, Ze-Jun Huo1, Hui-Feng Hao6, Lei Yang4, Chun-Shui Pan3,6, Ke He3,6, Jing-Yu Fan3,6, Zhi-Gang Li1 and Jing-Yan Han3,5,6

1 School of Acupuncture and Moxibustion, Beijing University of Chinese Medicine, Beijing, China 2 Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, Beijing, China 3 Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, China 4 Department of Anatomy, School of Basic Medical Sciences, Peking University, Beijing, China 5 Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, China 6 Key Laboratory of Microcirculation, State Administration of Traditional Chinese Medicine of the People's Republic of China This study was designed to investigate the effect of long-term electroacupuncture at Baihui

(DU20) and Zusanli (ST36) on cerebral microvessels and neurons in CA1 region of

hippocampus in spontaneously hypertensive rats (SHR). A total of 45 male Wistar rats and 45

SHR were randomly grouped, with or without electroacupuncture at DU20 and ST36 once

every other day for a period of 8 weeks. The mean arterial pressure (MAP) was measured once

every 2 weeks. Cerebral blood flow (CBF) and the number of open microvessels in

hippocampal CA1 region were detected by Laser Doppler and immunohistochemistry,

respectively. Nissl staining and western blotting were performed to determine hippocampus

morphology. The results showed that the MAP in SHR increased linearly over the observation

period and significantly reduced following electroacupuncture as compared with Wistar rats.

The CBF, learning and memory capacity of SHR were improved by electroacupuncture. The

capillary rarefaction was improved and the up-regulation of angiotension II type I receptor

(AT1R), endothelin receptor (ETAR) and endothelin-1 (ET-1) in SHR rats was attenuated by

electroacupuncture, suggesting an implication of AT1R, ETAR and ET-1 pathway in the effect

of electroacupuncture.

P7-2 Adoptive regulatory T-Cell therapy prevents cerebral vasospasm and reduces early brain injury following experimental SAH Li-Ping Zhang, Yan Cheng, Lei Zhang, Ming-Feng Yang, Zong-Yong Zhang, Feng-Ze Wang, Bao-Liang Sun

Department of Neurology and Key Lab oratory of Cerebral Microcirculation, Taishan Medical University, Taian 271000, Shandong, China Objective: Recent evidence suggests regulatory T cells (Tregs), an innate immunomodulator, have Immunosuppressive effects. Besides, immune response and inflammation response play critical role in the development of cerebral vasospasm (CVS) and early brain injury after experimental subarachnoid hemorrhage (SAH). We, therefore, evaluated the effects of Tregs transfer in rat SAH models and further investigated the mechanism underlying Treg-afforded neuroprotection. Methods: Wistar rats were divided into four groups: sham group (time matched control, n=6), PBS+SAH group (n=6), SP+SAH Group (n=6) and Tregs+SAH group (n=6). The rats in PBS+SAH group were injected intravenously 0.2ml PBS one hour after SAH. The rats in SP+SAH group were given 2x106 splenocytes in 0.2ml PBS. The rats in Tregs+SAH group received 2x106 Tregs in 0.2ml PBS. The degree of vasospasm was determined by averaging the cross-sectional areas and vascular wall thickness of the basilar artery 3 days after SAH. The integrity of blood-brain barrier could be disrupted by extracellular matrix metalloproteinase 9 (MMP9), therefore, we examined the degree of disruption of the BBB by the expression of MMP9. Levels of MMP9 were studied in brain tissue by immunofluorescence staining and western blot. The expression of apoptotic indicators, including active caspase-3 and terminal deoxy nucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL), indicated the degree of brain damage in SAH model. Western blot and immunofluorescence were used for quantification and location of active caspase-3. Neuronal cell death was quantified with TUNEL staining. Results: Intravenous administration of purified Tregs at 1 hour after experimental SAH attenuated CVS and improve neurological function of SAH rat model. Treg-afforded neuroprotection was accompanied by attenuated blood–brain barrier (BBB) disruption during early stages of SAH and decreased cerebral injury. It was found that the wall thickness of basilar artery in PBS+SAH group and SP+SAH group was significantly increased than that in control group (sham group), whereas the wall thickness of basilar artery is decreased markedly in Tregs+SAH group than that in PBS+SAH and SP+SAH group. In addition, the cross-sectional areas of basilar artery in PBS+SAH group and SP+SAH group was significantly increased than that in control group (sham group), whereas the cross-sectional areas of basilar artery in Tregs+SAH group is decreased markedly, as compared with PBS+SAH and SP+SAH group. Immunofluorescence staining and western blot demonstrated a significant increase in MMP9 in PBS+SAH and SP+SAH group, and Tregs treatment abolished the SAH-induced MMP9 elevation almost to sham levels. Tregs treatment significantly decreased expression of active caspase-3, compared with PBS+SAH group and SP+SAH group. It indicated that Tregs treatment significantly reduced neuronal cell death after SAH. Conclusion: Tregs treatment attenuates CVS, prevents the disruption of BBB, decreased neuronal cell death and improved neurological status in a rat model of SAH. Our study suggests that Tregs transfer therapy is a novel and potential cell-based therapy targeting early brain injury and cerebral vasospasm after SAH.

P7-3 Intravenous transplantation of In vitro expanded CD4+CD25+ regulatory T cells exerts protective effects and promotes the expression of Iba1, GFAP after cerebral Ischemia-Reperfusion in rats Yan Cheng, Li-Ping Zhang, Lei Zhang, Ming-Feng Yang, Zong-Yong Zhang, Feng-Ze Wang, Bao-Liang Sun Key Lab of Cerebral Microcirculation in Universities of Shandong (Taishan Medical University); Department of Neurology, Affiliated Hospital of Taishan Medical University, Taian, Shandong 271000, China Objective: Iba1 and GFAP are specifically expressed in microglia and astrocytes. Ischemia-Reperfusion injury is associated with the proliferation of microglia and astrocytes. In the present study, we sought to explore the protective effects of intravenous transplantation of In vitro expanded CD4+CD25+ regulatory T cells (tregs) and the influence on the expression of Iba1 and GFAP after cerebral ischemia-reperfusion in rats. Methods: Two-steps method of Magnetic cell sorting (MACS) was used to extract CD4+CD25+ regulatory T cells from healthy adult Sprague-Dawley (SD) rats. The Tregs were expanded in vitro in response to anti-CD3-anti-CD28-coated microbeads and IL-2. Flow cytometry was used to analyze the purity of the Tregs. Trypan blue staining was used to demine the cell survival. Rat middle cerebral artery occlusion was induced by the intraluminal filament technique. The middle cerebral artery was occluded for 2h, followed by reperfusion. The rats were divided randomly into normal group, sham-operation group, and MCAO group (further divided into PBS treatment group, Splenocytes treatment group (SP) and Tregs treatment group). Rat received femoral vein injection of 2×106 Tregs or Splenocytes diluted in 0.2ml PBS in Tregs and SP groups, while equal volume of PBS was injected via the femoral vein in PBS group. Neurological behavior was evaluated by Longa’s scoring method. TTC staining was applied to evaluate the infarct volume. Immunohistochemistry and western blotting were used to observe the expression of Iba1 and GFAP protein in the ischemic brain tissue at different time of ischemia and reperfusion. Results: The purity of CD4+CD25+Treg cells obtained by MACS separation was 84.50%, the cell survival rate was 95.00%. After 3 weeks of culture, the purity of Tregs was 76.00%, the cell survival rate was 94.30%. Neurological deficits after modeling were peaked at 3d (P﹤0.05, compared with 7d), neurological function of Tregs treatment group improved significantly as compared with the PBS treatment group and SP treatment group at 3d and 7d (P﹤0.01, respectively). The infarct area of Tregs treatment group reduced significantly compared with the PBS treatment group and SP treatment group at 3d and 7d (P﹤0.01, respectively). In the normal and sham groups, there were scattered Iba1positive cells, while PBS and SP groups appeared numerous Iba1 positive cells at 3d after reperfusion in the peri-ischemia area. The immunoreactivity was further increased and peaked at 7d. Iba1 positive cells of Tregs treatment decreased significantly compared with PBS and SP treatment groups (P﹤0.01). Similarly, the number of GFAP positive cells increased at 3d after ischemia-reperfusion, and continued until 7 days. The expression of GFAP protein in Tregs treatment group reduced significantly compared with PBS and SP treatment groups (P﹤0.01). The results of western blotting for Iba1 and GFAP were similar to the immunohistochemical data. Conclusion: The In vitro expanded CD4+CD25+ regulatory T cells transplantation significantly decreased the cerebral infarct volume and improved the neurological function. Our findings suggested that microglial and astrocytes are targets for CD4+CD25+ regulatory T cells, and that modulation of glial response may be a mechanism of Tregs-mediated neuroprotection in ischemia-reperfusion.

P7-4 Anti-inflammation treatment after subarachnoid hemorrhage: Regulatory T-cells adoptive transfer Lei Zhang, Li-Ping Zhang, Yan Cheng, Ming-FengYang, Zong-Yong Zhang, Feng-Ze Wang, Bao-Liang Sun Key Lab of Cerebral Microcirculation in Universities of Shandong (Taishan Medical University); Department of Neurology, Affiliated Hospital of Taishan Medical University, Taian, Shandong 271000, China Objective: Inflammation and immunity play a vital role in the pathogenesis of brain injury after subarachnoid hemorrhage (SAH). Nuclear factor-kappa B (NF-κB) regulates many genes essential for inflammation and immunity and is activated by toll-like receptor (TLR). Regulatory T cells (Tregs) play a critical role in maintenance of immunologic self-tolerance as well as regulation of immune responses. This study aimed to detect the protective effects of Regulatory T-cells and the influence on the toll-like receptor 4/nuclear factor-kappa B (TLR4/NF-κB) signaling in the rat brain after SAH. Method: Regulatory T-cells were obtained from spleen of rat using immunomagnetic beads. Rat SAH models were replicated by a modified cistern magna injection with autologous arterial hemolysates. The rats were divided into the normal group, sham-operated control group (sham), and SAH group. Rats in SAH group were divided into PBS treated group, SP treated group and Tregs treated group. Seventy-two hours after SAH, the brain was prepared for paraffin sections and fresh sections. Immunohistochemical staining of TLR4 in brain tissue was carried out. Western blotting was used to evaluate the expression of NF-κB and IL-6. Result: Compared with the normal control group and sham group, intensive TLR4-like immunostaining was present extensively in the brain after SAH. At the same time, the level of NF-κB and IL-6 in SAH group was significantly increased. The level of TLR4, NF-κB and IL-6 in the brain tissue in Tregs treated group was significantly lower, compared with the PBS treated group and SP treated group. Conclusion: We have characterized the expression pattern of proinflammatory cytokines and TLR4 with a link of NF-κB signaling in the brain injury after experimental SAH. Inflammatory responses mediated mainly by the activation of the TLR4/NF-κB signaling pathway may play an important role in the pathogenesis of early brain injury following SAH. Regulatory T-cells transplantation significantly reduce the level of TLR4, NF-κB and IL-6 in the brain tissue. It suggested that Regulatory T-cells transplantation attenuates brain damage after SAH by limiting the inflammation in the central nervous system.

P7-5 Blocking neurodegenerative damages of Alzheimer's disease by aromatic glycosides in multiple pathways

Ying-Hong Wang a, Shao-Yang Zhao a, Ke Zhang b, Xu Ma b, Hai-Xia Li b, Shu-Yan Han b,

Yong Jiang b, Hou-Nan Wu c, Zhuo-Ping Huo d, Peng-Fei Tu b* , Zhi-Zhong Ma a*

a Department of Combined Traditional Chinese Medicine with Western Medicine, School of

Basic Medical Science, Peking University, Beijing 100191, China b Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University,

No.38. Xueyuan Road, Beijing 100191, China c Medical and Healthy Analytical Center, Peking University, No.38. Xueyuan Road, Beijing

100191, China c Institute of Child and Adolescent Health, School of public health, Peking University, No.38.

Xueyuan Road, Beijing 100191, China

* Corresponding authors

Blocking neurodegeneration of Alzheimer’s diseases is global puzzles in medicine. Single

blocking either oxidative stress or inflammation is not ideal strategy for this complicated

illness. Through our studies, we found some of phenolic glycosides (Xy-B-25, Xy-B-23,

BL-B-14, BL-E-8, SCI-Et-18, SC2-19 and IO-B-40) present significant inhibition on cortical

neuronal death induced by Aβ1-42. Simultaneously, the endogenous ROS, biomarker of DNA,

lipid and protein damages were all reduced dramatically. The ROS induced Aβ1-42 production,

as well as p-38MAPK phosphorylation, were markedly deduced. Additionally, the activation of

microglia and astrocyte, their releasing of inflammatory cytokines of COX-1, COX-2, IL-1β,

TNFα, as well as p-38MAPK phosphorylation were inhibited. In SAM-P8 mice, which is

expressed premature aging, senile demention and multiple neurodegenerations, were tested and

the results showed that one of our compounds was effective in improving the learning and

memory ability as well as reducing Aβ1-42 production and p-38MAPK phosphorylation.

Keywords: Alzheimer’s disease, phenolic glycosides, Oxidative stress, inflammation,

neurodegeneration

P7-6

The involvement of Programmed Cell Death 5（PDCD5）in the regulation of

apoptosis in cerebral ischemia/reperfusion injury

Chun-Hua Chen,1 Zhao Jiang,1 Xiao-Mei Yang,1 Jun-Hao Yan,1 Lei Yang,1 Ke Wang,1 Ying-Yu

Chen,2 Chang-Man Zhou1*

1Department of Anatomy and Embryology, Peking University Health Science Center, Beijing,

China

2Peking University Center for Human Disease Genomics, Beijing, China

Introduction: Programmed Cell Death 5 (PDCD5) is a protein that accelerates apoptosis in

different types of cells in response to various stimuli, and is down-regulated in many cancer

tissues.

Aims: In this study, we found for the first time that PDCD5 plays an essential role in brain

ischemic injury in a rat focal cerebral ischemia model. We hypothesized that down-regulating

PDCD5 can protect the brain from ischemic damage by inhibiting PDCD5 induced apoptotic

pathway.

Results: The PDCD5 siRNA reduced the infarct volume, improved neurological deficits and

reduced Evans blue extravasation. Meanwhile, over-expression of PDCD5 protein using

recombinant human PDCD5 (rhPDCD5) had the opposite effect. But unfortunately, changed

PDCD5 expression had no significant effect on mortality. Immunohistochemistry and western

blot demonstrated PDCD5 siRNA decreased expressions of key pro-apoptotic proteins such as

p53, Bax/Bcl-2 and cleaved casepase-3 in the infarcted areas, whereas PDCD5 over-expression

increases cell apoptosis.Double fluorescence labeling showed PDCD5 positive immunoreactive

materials were partly colocalized with MAP2, GFAP, p53 and caspase-3 in the injured cerebral

cortex.

Conclusions: PDCD5 induced apoptosis and over-expression of PDCD5 is harmful to the

ischemic neurons in vivo. Meanwhile, inhibition of PDCD5 may be protective via reducing the

apoptotic-related protein such as p53, Bax and caspase-3. This observation may have potential

for the treatment of ischemic cerebral stroke.

Poster 8 Clinic and microcirculation

Chair: Hui Cai

Department of Integrated Traditional and Western Medicine, Nanjing General Hospital of Nanjing Military Region PLA

13:20-13:30 P8-1 The correlation analysis between the visualization of hemorrheologic changes and platelet function on the patients with acute coronary syndrome after percutaneous coronary intervention Jia-Qi Wang Xiyuan Hospital, China Academy of Traditional Chinese Medicine 13:30-13:40 P8-2 To observe the effect of Tong-Mai-Yang-Xin pill in adjuvant treatment of three kinds of blood diseases Ai-Hong Wei The 210th Hospital of PLA, TCM Hematology Center 13:40-13:50 P8-3 Analysis on the thinking of using non-traditional promoting blood circulation to remove blood stasis drugs to improve the microcirculation Wan-Li Gu Department of traditional Chinese medicine, Liaocheng People's Hospital 13:50-14:00 P8-4 Study for the treatment of integrative traditional Chinese and western medicine on acute-on-chronic or subacute-on-chronic liver failure in a multiregional randomized and controlled trial Jun Li The 302 military hospital of P.L.A 14:00-14:10 P8-5 Hemorheology visualization clinical research of elderly hypertension with different TCM syndromes Xiong Wei Xiyuan Hospital, China Academy of Chinese Medical Sciences 14:10-14:20 P8-6 Rhubarb Root and Rhizome-based Chinese herbal prescriptions for acute ischemic stroke: a systematic review and meta-analysis of 12 randomized controlled trials Guo-Qing Zheng The Center of Neurology and Rehabilitation, The Second Affiliated Hospital of Wenzhou Medical University

P8-1 The correlation analysis between the visualization of hemorrheologic changes and platelet function on the patients with acute coronary syndrome after percutaneous coronary intervention

Jia-Qi Wang, Jian-Gang Liu, Cheng-Long Wang Xiyuan Hospital, China Academy of Traditional Chinese Medicine, Beijing100091

AIM: The purpose of this study was to intuitionally observe the blood rheology characteristics on the patients with acute coronary syndrome (ACS) after percutaneous coronary intervention (PCI) for one year to three years by Micro Channel array Flow Analyzer (MC-FAN) combining with other platelet function indices, and to explore the correlations between the test results of MC-FAN and platelet function. METHODS: This study brought into 74 patients with ACS after PCI for one year to three years as test group, 21 healthy subjects as normal group, no drug intervention, detected the PAgT, PAdT, P-selection, PDGF-BB, vWF, and used MC-FAN HR300 to detect the transiting time (MC-FAN TT) of the in vitro blood passing through the model body capillaries. This study compared the differences of the test results between two groups, and explored the correlations between the test results of MC-FAN and platelet function on the patients with ACS after PCI. RESULTS: In the test group, compared with normal group, the MC-FAN TT were longer (P<0.01), the ability of erythrocyte deformation weakened, leucocyte attaching the vascular wall and platelet adhering and gathering relatively increased. The levels of PAgT, PAdT, P-selection, PDGF-BB in test group were all higher than normal group (P<0.01). The difference of vWF between two groups had no statistical significance (P>0.05). The intergroup correlations analysis showed: There were correlations between MC-FAN TT and platelet function. 10μl TT, 30μl TT had the most significant correlation with P-selection (r=0.601, P=0.000; r=0.334, P=0.009); 60μl TT had the most significant correlation with PAgT (r=0.527, P=0.007); 100 μl TT had the most significant correlation with PAdT (r=0.815, P=0.014). CONCLUSION: The visualization of hemorrheologic changes and platelet function on the patients with ACS after PCI are abnormal, which need antiplatelet therapy for a long time. MC-FAN can intuitionally observe the patients’ changes of blood rheology and objectively evaluate the condition of the patients’ blood rheology. There are correlations between MC-FAN TT and platelet function.

P8-2 To observe the effect of Tong-Mai-Yang-Xin pill in adjuvant treatment of three kinds of blood diseases Ai-Hong Wei, Xiao-Hui Chang The 210th Hospital of PLA, TCM Hematology Center, Dalian 116021

Objective: To observe the effect of Tong-Mai-Yang-Xin pill (TMYX) in adjuvant treatment of chronic regeneration barrier anemia (AA) with microcirculation disorder, idiopathic thrombocytopenic purpura (ITP), iron deficiency anemia (IDA), to alleviate the patients symptoms such as palpitations shortness of breath, improve heart function, alleviate microcirculation, improve patient quality of life. Methods: 60 cases were clinic patients, average age 50 + 21 years old, male 11, female 49 cases. In 25 patients with AA, 29 patients with ITP, 6 patients with IDA. In 60 cases with hypertension in 18 cases, 27 cases of coronary heart disease, 10 cases of gastrointestinal diseases, 9 cases of climacteric syndrome, 5 cases of diabetes, 2 patients with lung injury, 3 cases of liver injury. Oral TMYX (china-singapore tianjin pharmaceutical) 40 pills per time, 2 times a day, AA patients with stanozolol (Nanning Guangxi Baihui pharmaceutical) 4mg per time, 2 times a day, after meals; ITP patients with transfer factor oral solution (Gannon Hailing pharmaceutical) 20ml per time, 2 times a day, after meals; IDA patients with ferrous sulfate folic acid tablets (Jolliness Pharmaceutical) 3 tablets per time, 2 times a day, meals. After two months treatment, observed clinical symptoms, blood routine, and tongue microcirculation. Results: Before treatment, 48 cases with Palpitations, shortness of breath, chest tightness; 19 cases with dry mouth, 6 cases with epigastric discomfort, 3 cases with abnormal liver function; 10 cases with tongue light; 22 cases with tongue red; 28 cases with tongue dim; 39 cases with sublingual venous engorgement; 48 with tongue microcirculation moderately severe obstacles; 27 cases with pulse condition heavy fine; 33 cases with pulse condition string slide. To observe all patents’ blood routine: in AA white blood cells (3.48 + 0.10) x 109 / L, hemoglobin (105.7 + 8.83 g/L, platelet (45.29 + 17.42) x 109 / L; in platelet (50.04 + 10.54) x109 /L; in IDA hemoglobin (73.24 + 21.17) g/L. After treatment, the Palpitations, shortness of breath, chest tightness was relieved in 46 cases, the recovery was 82.76%; dry mouth was relieved in 16 cases, the recovery was 89.5%; the epigastric discomfort was relieved in 4 cases, the recovery was 66.7%. Abnormal hepatic function recovery rate was 100%. Pulse condition recovery rate was 89.1%; the tongue recovery rate was 77.59%; sublingual venous engorgement recovery rate was 62.07%; the tongue microcirculation recovery rate was 48.5%. To observe all patents’ blood routine: in AA white blood cells (3.62±0.11) x 109 / L, hemoglobin (107.41±8.16) g/L, platelet (44.70±15.78) x 109 / L; in platelet (48.85±13.29) x109 /L; in IDA hemoglobin (135.50±31.74) g/L. Conclusion: TMYX consists of 11 Chinese herbs, such as radix rehmanniae, radix glycyrrhizae, dangshen, fleece-flower root, dwarf lilyturf, fructus schisandrae, donkey-hide gelatin, tortoiseshell, caulis spatholobi, cassia twig, big jujube and is attending coronary heart disease and arrhythmia. Because of consisting the tortoise shell, radix polygoni multiflori, radix rehmanniae, caulis spatholobi, fructus schisandrae, which have the ziyin yangxue efficacy. We choose it for the treatment of blood diseases with microcirculation disorder. After 2 months treatment, the symptoms has improved significantly for blood disease accompanied with hypertension, coronary heart disease, gastrointestinal disease, menopausal syndrome, diabetes, lung injury, liver injury .The routine blood was relatively stable for AA patients, no elevated role for ITP. The hemoglobin was elevated for IDA patients. TMYX was safety for the above three kinds blood diseases, had adjuvant treatment affection, could improve the microcirculation and recovered the abnormal liver function.

P8-3

Analysis on the thinking of using non-traditional promoting blood circulation to remove blood stasis drugs to improve the microcirculation Wan-Li Gu Department of traditional Chinese medicine, Liaocheng People's Hospital, Shandong, 252000 Objective: To explore the application of research ideas of non traditional promoting blood circulation to remove blood stasis drugs improving microcirculation disorder. Methods: The traditional Chinese medicine has "Chronic illness into the network becomes blood stasis" theory, and blood stasis patients generally have the performance of microcirculation disorder, such as micro slow blood flow and blood stasis, and intravascular coagulation, micro vascular deformation (pipe loop twisted, deformity, the top expansion), microvascular peripheral blood oozing and bleeding, vascular contraction narrow or occlusion. Through the review of the traditional definition, promoting blood circulation to remove blood stasis type and classification, with angina pectoris of coronary heart disease as an example, from the Chinese medicine theory of Qi and blood, blood stasis syndrome, the theory of five viscera correlation analysis on non-traditional promoting blood circulation to remove blood stasis drugs, promoting blood circulation to remove blood stasis and improve microcirculation function are discussed. Results: The common non-traditional promoting blood circulation to remove blood stasis drugs include: Qi invigorating drugs (born licorice, astragalus, Codonopsis, Atractylodes rhizome), nourishing the blood medicine (radix paeoniae alba), tonifying yang drug (epimedium), nourishing Yin drug (Rehmannia glutinosa), the divergent cold medicine (cassia twig), Heat clearing and detoxifying drugs (Scutellaria baicalensis), resolving hard lump drug (seaweed), dampness medicine (Poria). The promoting blood circulation to remove blood stasis role there is a record of past dynasties literature, but the modern clinical neglected. Dialectical using non-traditional promoting blood circulation to remove blood stasis drugs can improve the micro circulation and obtain better curative effect. Conclusion: Appropriately expand the scope of drugs promoting blood circulation and removing blood stasis, expand the scope of application of drugs to improve microcirculation, can open microcirculation research field and deepen the study of microcirculation.

P8-4

Study for the treatment of integrative traditional Chinese and western medicine on acute-on-chronic or subacute-on-chronic liver failure in a multiregional randomized and controlled trial Jun Li The 302 military hospital of P.L.A, Beijing, 100039

Aim: To make sure of the change of fatality rate by the complicated treatment of integrative

traditional Chinese and western medicine on acute-on-chronic or subacute-on-chronic liver

failure in the early stage of disease.

Methods: The designs in this trial were multiregional randomized and controlled. The

randomly proportion of the amount of the patients that were separated into two groups either

western medicine or integrated traditional Chinese were 1:2. We gave them the physical and

laboratory examination at the 8th, 12th, 24th, 48th week.

Results: By CMH chi-squared test, it appears that the difference between the two groups is

exactly. The survival analysis shows that fatality rate in the integrated group is lower than that

in the western group. The test of homogeneity between the two groups’ survival curve shows

that the survival time in the integrated group is longer than that in the western group (p＜0.05).

Conclusion: The fatality rate could be degraded and the survival rate could be advanced

through the complicated treatment of western integrated traditional Chinese medicine on

acute-on-chronic or subacute-on-chronic liver failure in the early stage of disease.

Key words: liver failure; the treatment of western integrated traditional Chinese medicine; the

fatality rate

P8-5 Hemorheology visualization clinical research of elderly hypertension with different TCM syndromes

Xiong Wei, Jian-Gang Liu, Hao Li Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091 Objective Apply micro channel array flow analyzer (MC-FAN) to observe blood flow characteristics of elderly hypertension patients with different Traditional Chinese Medicine syndrome and testing the correlation between them and red blood cell rheological property. Methods Selected 109 elderly hypertension patients, and according to Traditional Chinese Medicine syndrome differentiation, divided them into phlegm-stasis blocking collateral group (35 cases), yin deficiency with yang hyperactivity group (42 cases), kidney deficiency group (32 cases). And there is a elderly healthy group (21 cases). Using MC-FAN to do the hemorheology visualization testing of elderly hypertension patients and observing the erythrocyte deformation indexes, erythrocyte aggregation indexes and erythrocyte related plasma ATPase activity and do the relevant analysis between micro channel array flow transiting times and red blood cell rheological indexes. Results Micro channel array flow transiting time comparison: Phlegm-stasis blocking collateral group compare with the micro channel array flow transiting times of yin deficiency with yang hyperactivity group and kidney deficiency group and elderly healthy group, the comparison of statistics has significant extended (P<0.05). When shear rate is 100 s-1, the erythrocyte deformation index comparison: phlegm-stasis blocking collateral group compare with yin deficiency with yang hyperactivity group and kidney deficiency group and elderly healthy group has significant reduced (P<0.05). Erythrocyte related plasma ATPase activity comparison: Plasma Na+-K+-ATPase activity and of Plasma Ca2+-Mg2+-ATPase activity of elderly hypertension group compare with elderly healthy group have significant reduced (P<0.01). Among the three groups, Plasma Ca2+-Mg2+-ATPase activity of phlegm-stasis blocking collateral group compare with kidney deficiency group compare has significant reduced (P<0.05). The correlation of elderly hypertension patients’ micro channel array flow transiting times and red blood cell rheological indexes: elderly hypertension patients’ micro channel array flow transiting times (10µL, 30µL, 60µL and 100µL) were respectively significant negative correlated with erythrocyte deformation index. 100µL transiting time was significant positive correlated with erythrocyte aggregation index. Conclusion Micro channel array flow transiting time of elderly hypertension patients has significantly changed. And it has the correlation with traditional erythrocyte rheological property.

P8-6

Rhubarb Root and Rhizome-based Chinese Herbal Prescriptions for Acute Ischemic Stroke: A Systematic Review and Meta-analysis of 12 Randomized Controlled Trials Guo-Qing Zheng*, Lin Lu, Deng-Lei Fu, Ling-Yan Lu, Ai-Ju Liu, Hui-Qin Li, Ji-Huang Li, Yuan-Yuan Wang The Center of Neurology and Rehabilitation, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325027, China Background: Traditional Chinese Medicine has been used in stroke victims for thousands of years, and the purging fu-viscera (Tongfu) method with rhubarb root and rhizome-based Chinese herbal prescriptions was one of the most typical and representative treatment principle for stroke from ancient to modern times. Rhubarb root and rhizome, its effective components and its prescriptions have been demonstrated to be beneficial effects on multi-aspects of the pathophysiology of stroke. The objective of this review is to assess clinical efficacy and safety of rhubarb root and rhizome-based prescriptions for stroke. Methods: A systematic literature search in six databases was performed to identify randomized controlled trials (RCTs) comparing rhubarb root and rhizome-based prescriptions with western conventional medicine (WCM) for acute ischemic stroke. The methodological quality of RCTs was assessed independently based on the 12 categories recommended by the Cochrane Back Review Group. Results: A total of 968 participants were included in 12 eligible studies. All trials were deemed to have a relative high risk of bias. Rhubarb root and rhizome-based prescriptions have a significant effect on improvement of Barthel Index scores (n=5), National Institutes of Health Stroke Scale scores (n=2), Glasgow Coma Scale scores (n=1), neurological deficit score (N=5), and the clinical efficacy rate (n=10) when compared with the WCM control (p <0.05 or p <0.01). Six trials reported that there were no adverse events, while not mention whether they monitored adverse effects at all in left 6 studies. Conclusions: Despite of the poor methodological quality, rhubarb root and rhizome-based prescription appeared to improve the clinical neurological impairments and seemed to be generally safe in patients with acute ischemic stroke. Rhubarb root and rhizome-based prescription as a promising candidate is worthy of improvement and development for further clinical stroke trials. Keywords: Acute ischemic stroke; Rhubarb root and rhizome; Tongfu method; Systematic review

参会名单序号姓名职称/职务单位

1 Erik Bruce Kistler

Assistant Professor

Department of Bioengineering, The Institute of Engineering in Medicine University of California San Diego

2 Masato Yasui

Professor and Chair

Department. of Pharmacology, School of Medicine, Keio University

3 Ying-Chun Yu

Doctor School of Medicine, Keio University

4 Sarah Yuan Professor and Chair

Department of Molecular Pharmacology & Physiology Bob and Evelyn Deriso Endowed Chair University of South Florida Morsani College of Medicine

5 David Zawieja

Regents Professor and Vice Chair

Dept. of Medical Physiology Director - Division of Lymphatic BiologyTexas A&M University Health Science Center, College of Medicine, Department of Medical Physiology and Division of Lymphatic Biology

6 Walter N Durán

Professor and Vice Chair

Pharmacology & Physiology Division of Vascular Surgery

7 Pierre Maechler

Professor Department of Cell Physiology and Metabolism, University of Geneva Medical Centre

8 Jerome W. Breslin

Associate Professor

Molecular Pharmacology and Physiology Morsani College of Medicine, University of South Florida

9 Mack Wu Professor of Surgery

University of South Florida College of Medicine

10 SuematsuMakoto

Professor and Dean

Department of Biochemistry and Integrative Medical Biology School of Medicine, Keio University

11 Yu-Ling Ma Ph.D. University of Oxford 12 韩晶岩教授、主任北大医学部中西医结合学系、北大医学

部天士力微循环研究中心

13 沈剑刚教授香港大学中医药学院

14 黄巧冰教授南方医科大学基础医学院病理生理教

研室

15 朱海波教授北京协和医学院药物研究所心脑血管

实验室

16 朱彦教授天津中医药大学

17 朱丹教授华中科技大学生命科学与技术学院，武

汉光电国家实验室

18 孙保亮教授泰山医学院

19 郭晓华讲师南方医科大学病生理教研室

20 周长满教授北大医学部基础医学院解剖学系

21 李学军教授北大医学部基础医学院

22 蔡辉教授南京军区南京总医院

23 谭睿教授西南交大生命学院

24 张宏权教授北大医学部基础医学院

25 戴克胜教授苏州大学

26 吴剑波中心主任，研究

员

四川省泸州医学院药物研究中心

27 魏勋斌教授上海交通大学

28 杜军保主任北京大学第一医院儿科

29 杨宝学教授、副主任北大医学部基础医学院药理学系

30 廖福龙教授中医研究院中药研究所药理室

31 霍则军副教授北京大学第三医院

32 张雷研究生西苑医院

33 陈兰玉研究生中日友好医院心内科

34 赵启韬副教授山东中医药大学基础医学院

35 魏艾红教授大连市解放军 210 医院中医血液科

36 谷万里主任医师聊城市人民医院

37 李筠教授中国人民解放军 302 医院

38 杨宁助理研究员北京军区联勤部药品仪器检验所

39 郑国庆教授温州医学院附属第二医院神经内科

40 Yi-Cun Wu 研究生苏州大学

41 Ming Li 研究生苏州大学

42 Jian-Sheng

Zhang

研究生苏州大学

43 王倩副主任医师首都医科大学附属北京中医医院心血

管科 44 金文副研究员中国医学科学院药用植物研究所

45 方晓艳副教授河南中医学院药学院 46 张晓云教授河北北方学院

47 张栋研究员中国中医研究院针灸研究所

48 陈彦静教授中国中医科学院中医基础理论研究所

实验中心生理室

49 任钧国副教授中国中医科学院西苑医院

50 徐明副研究员北京大学第三医院血管医学研究所

51 王丹巧副教授中国中医科学院医学实验中心

52 张学智主任，主任医师北京大学第一医院中医中西医结合科

53 刘红旭主任，主任医师首都医科大学附属北京中医医院

54 韩东主任，副研究员国家纳米科学中心

55 金明主任,主任医师卫生部中日友好医院眼科

56 刘育英教授北京大学医学部天士力微循环研究中

心

57 古川修访问学者北京大学医学部天士力微循环研究中

心

58 罗茂助理研究员四川省泸州医学院药物研究中心

59 张立民研究生河北北方学院

60 杜会波研究生河北北方学院

61 韩波研究生河北北方学院

62 张玉萍研究生河北北方学院

63 李蓉助理研究员四川省泸州医学院药物研究中心

64 常晓慧主治医师大连市解放军 210 医院中医血液科

65 魏睦新中医科主任、教

授

南京医科大学中西医结合研究所

南京医科大学第一附属医院

66 余上斌教授武汉同济医学院机能学中心实验室

67 张礼萍研究生泰山医学院脑科研究所

68 程焱研究生泰山医学院脑科研究所

69 张磊研究生泰山医学院脑科研究所

70 张丽莙教授天津医科大学基础医学院病理生理教

研室 71 Jun‐Yan Wei 研究生天津医科大学基础医学院病理生理教

研室 72 吴丽丽

研究生南方医科大学基础医学院病理生理教

研室

73 黄力教授中日友好医院中西医结合心内科

74 孙桂波教授中国医学科学院药用植物研究所药理

室

75 秦合伟上海中医药大学附属龙华医院

76 赵丁教授河北医科大学药学院生药室

77 郝璋森硕士研究生河北医科大学药学院生药室

78 秦鉴主任医师、博导

教育处处长

中山大学附属第一医院

79 刘萍处长、研究员上海中医药大学附属龙华医院人事处

80 李艳彦副教授山西中医学院

81 王永辉副教授山西中医学院

82 赵自刚教授河北北方学院

83 牛春雨教授河北北方学院

84 黄怀宇科主任，主任医

师

江苏省南通市第一人民医院神经内科

85 薛欣副研究员中国中医科学院基础理论研究所

86 施秋萍研究生北京大学第一医院心内科

87 叶菲教授中国医学科学院药物研究所

88 张晓玲研究生中国医学科学院药物研究所

89 徐筱杰教授北大化学院

90 陈春花副教授北京大学医学部基础医学院解剖教研

室

91 杨磊副主任技师北京大学医学部基础医学院解剖教研

室

92 范颖副院长、教授辽宁中医药大学基础医学院

93 霍勇教授北京大学第一医院

94 周亚伟教授北大世佳研究中心

95 赵智明教授南京军区南京总医院

96 于春泉主任、副研究员天津中医药大学期刊编辑部

97 高杉编辑天津中医药大学期刊编辑部

98 李琳编辑天津中医药大学期刊编辑部

99 秦大莲教授四川省泸州医学院药学院药理教研室

100 宁德科药师大连市解放军 210 医院中医血液科

101 王铭研究生中国中医科学院西苑医院

102 雷燕教授中国中医科学院西苑医院

103 姚魁武主任医师中国中医科学院广安门医院医教处

104 张鑫月研究生河北医科大学附属以岭医院

105 龙铟副主任医师第四军医大学西京医院中医科

106 赵树华主任,教授吉林大学中日联谊医院中医科

107 武俏丽天津市环湖医院

108 李小娟科主任辽宁中医附属医院

109 唐浩硕士研究生哈尔滨医科大学

110 刘端勇副教授江西中医学院

111 芦瑀住院医师北京王府中西医结合医院

112 姜旭光副教授山东高等医学专科学校

113 李彦川助理研究员天士力集团研究院药理所

114 林色奇副教授江西中医学院

115 黄慧玲教授天津环湖医院

116 白文主任医师北京大学人民医院中医科

117 熊伟研究生西苑医院

118 刘剑刚教授西苑医院

119 高天舒科主任辽宁中医附属医院

120 王家琪研究生西苑医院

121 张蕾研究生西苑医院

122 杨晓媛博士研究生北大医学部中西医结合教研室

123 李翀硕士研究生北大医学部中西医结合教研室

124 田贵华主治医师东直门医院

125 李红娟硕士研究生东直门医院

126 阎本永博士研究生北京大学第一医院中西医结合科

127 李琳博士研究生卫生部中日友好医院中西医结合心内

科

128 陈福勤硕士研究生北京大学人民医院中医科

129 潘春水助理研究员北大医学部天士力微循环研究中心

130 阎丽助理研究员北大医学部天士力微循环研究中心

131 孙凯助理研究员北大医学部天士力微循环研究中心

132 李泉助理研究员北大医学部天士力微循环研究中心

133 胡白和室主任北大医学部天士力微循环研究中心

134 常昕主管技师北京大学医学部医药卫生分析中心

135 赵新荣主管技师北京大学医学部医药卫生分析中心

136 王传社副教授北大医学部天士力微循环研究中心 137 李娜主任秘书北大医学部天士力微循环研究中心 138 曲玫颖中心秘书北大医学部天士力微循环研究中心 139 王明暇助理研究员北大医学部天士力微循环研究中心

141 黄娉助理研究员北大医学部天士力微循环研究中心

142 马丽千博士研究生北大医学部中西医结合教研室

143 李志新副教授北大医学部中西医结合教研室

145 马治中副教授北大医学部中西医结合教研室

146 赵少阳硕士研究生北大医学部中西医结合教研室

147 王英红硕士研究生北大医学部中西医结合教研室

148 黄丹丹硕士研究生北大医学部中西医结合教研室

149 毛小伟博士研究生北大医学部中西医结合教研室 150 黄丹丹硕士研究生北大医学部中西医结合教研室 151 牟洪娜博士研究生北大医学部中西医结合教研室 152 崔元辰博士研究生北大医学部中西医结合教研室 153 贺珂博士研究生北大医学部中西医结合教研室 154 陈媛媛博士研究生北大医学部中西医结合教研室 155 肖萌萌硕士研究生北大医学部中西医结合教研室 156 孙昊煜博士研究生北大医学部中西医结合教研室 157 郝会峰博士研究生北大医学部中西医结合教研室 158 肖瑞平所长、教授北京大学分子医学研究所

159 王珏教授北京大学分子医学研究所

160 胡耀豪副教授北京大学分子医学研究所

The Professional Committee for Microcirculation, Chinese

Association of Integrative Medicine

number Name Positions and Technical

Titles

Work unit

1 Jing-yan Han

Director,

professor

Department of the Integration Chinese and Western Medicine,/Tasly Microcirculation Research Center, Peking University Health Science Center

2 Qiao-bing Huang

Professor Department of physiological pathology, School of Basic Medical Sciences, Southern Medical University

3 Yu-ying Liu

Professor Tasly Microcirculation Research Center, Peking University Health Science Center

4 Li Huang Director,

archiater

Combine traditional Chinese and western medicine of cardiology, China-Japan friendship hospital

5 Zhong-min Liu

Professor Department of Thoracic Surgery of Tongji University dongfang hospital

6 Xue-qi Li President,

Professor

Harbin Medical University fourth hospital

7 Da-zhuo Shi

Researcher Xiyuan Hospital CACMS

8 Wei-kang Wu

professor Research institution of the Integration Chinese and Western Medicine, Faculty of medical sciences, Sun YAT-SEN University

9 Chuan-she Wang

Associate

professor

Department of the Integration Chinese and Western Medicine, School of Basic Medical Sciences, Peking University；Tasly Microcirculation Research Center, Peking University Health Science Center

10 Hui Cai Director,

Professor

Department of traditional Chinese medicine, Nanjing General hospital of Nanjing Military Command

11 Shi-jun Wang

President,

Professor

School of Basic Medical Sciences ,Shandong University of traditional Chinese medicine

12 Chang-yi Guan

archiater Xiamen hospital of T.C.M, Fujian University of traditional Chinese medicine

13 Gui-yuan

Lv

Professor Department of Materia Medica ,Zhejiang Chinese medical University

14 Yan Lei Professor Research management of CACMS 15 Ming Jin archiater ophthalmology department of China-Japan friendship

hospital 16 Ming-yu President Thrombosis combine traditional Chinese and western

Chi medicine medical center in liaoning province 17 Ming-qi

Gao Professor China medical university clinical medical college of basic

medical center, 18 Shu-hua

Zhao Archiater Professor

Department of traditional Chinese medicine, JILIN University of China-Japan friendship hospital

19 Mao-yuan Wei

professor Combine traditional Chinese and western medicine research institute in heilongjiang province

20 Ding-fang Cai

Professor Department of the Integration Chinese and Western Medicine of Neurological research , Zhongshan hospital of FUDAN University

21 Qing-fu Zhang

Professor Burn and Plastic Surgery，The first hospital, hebei medical university

22 Chun-yu Niu

Professor Hebei North University

23 Guang-ying Huang

Professor Department of the Integration Chinese and Western Medicine ，Tongji hospital, huazhong university of science and technology

24 Mu-xin Wei

Professor The first affiliated hospital of nanjing medical university

25 Jin-shui Chen

Professor The first affiliated hospital of fujian medical university

26 Ke-sheng Dai

Professor Beijing University of Aeronautics and Astronautics

27 Dong Han

Director, associate

professor

National Center for Nanoscience and Technology

28 Jun Li Archiater The people's liberation army 302th hospital

29 Hong-xu Liu

Archiater Beijing hospital of traditional Chinese medicine, Capital medical university

30 Zhen-huan Liu

Archiater, professor

Nanhai maternity and child healthcare hospital,Guangzhou university of Chinese medicine

31 Yi-qun Zhang

Archiater, Department of the Integration Chinese and Western Medicine ,Jiangxi province people's hospital

32 Bao-liang Sun

Professor Neurology Department of Mount taishan medical school affiliated hospital

33 Guang-xian Cai

Professor Hunan University Of Chinese Medicine

34 Zhong-ming Xie

Professor Wuhan university people's hospital

35 Xue-zhi Zhang

Archiater department of traditional Chinese medicine combine traditional Chinese and western medicine ,Peking University first hospital

36 Yan-yan Li

Associate

professor

Shanxi college of traditional Chinese medicine

37 Ai-hong Wei

Associate chief physician

Dalian 210 hospital

38 Guang-li Wu

Professor department of nephropathy ,ethune international peace hospital of Beijing military area command

39 Yue-feng Liu


First affiliated hospital of chongqing medical university

40 Qing-hui Qi

Subdecanal The first hospital affiliated to dalian medical university

41 Rui Liang Associate

chief physician

The second hospital affiliated to dalian medical university

42 Wan-li

Gu


Liaocheng city people's hospital

43 Lin-shan

Jia

Combine traditional Chinese and western medicine journal of cardio-cerebrovascular disease

44 Jian-ping

Li

Associate professor

Henan university of Chinese medicine

45 Su-hong

Chen

Associate professor

department of traditional Chinese medicine, wenzhou medical college

46 Dian-qiao

Wang

Associate professor

Experimental reseach center China academy of Chinese medical sciences,

47 Jun-guo

Ren

Associate professor

Xiyuan Hospital CACMS

48 Ming Xu Associate professor

Department of Vascular Medicine, Peking University third hospital

49 Gui-bo

Sun

Associate professor

The Institute of Medicinal Plant Development

50 Kui-wu

Yao


Guang’anmen hospital, China academy of Chinese medical

sciences

51 Yan-jing Chen

Professor Institute of basic theory of TCM, China academy of Chinese medical sciences

52 Zi-gang Zhao

Professor Hebei North University/ Institute of microcirculation

53 Dong Zhang

Researcher Institute of acupuncture and moxibustion, China academy of Chinese medical sciences

54 Fei Ye Researcher Institute of Materia Medica, China academy of Chinese medical sciences

55 Li-jun Zhang

Professor Pathological physiology teaching and research section, School of Basic Medical Sciences, Tianjin Medical University

56 Hui-ling Huang

Professor Tianjin around the hospital

57 Ping Liu Professor Longhua hospital of Shanghai university of TCM

58 Hai-bo Zhu

Professor Institute of Materia Medica, China academy of Chinese medical sciences

59 Jian-gang Liu

Associate professor

Xiyuan Hospital CACMS

60 Xiao-dong Lv

Professor The first affiliated hospital of liaoning medical university

61 Li Cao Professor The Institute of Medicinal Plant Development 62 Xiao-jian

Xia Associate chief physician

Jiangxi province people's hospital

63 Xiu-rong Gao

Archiater Cadres three wards, The first hospital of Harbin

64 Wen Bai Archiater department of traditional Chinese medicine , Peking University People's Hospital

65 Ding Zhao

Professor hebei medical university

66 Jia-yi Cheng

Associate professor

School of Basic Medical Sciences, Liaoning university of traditional Chinese medicine

67 Ying Fan professor Liaoning university of traditional Chinese medicine

68 Xin Liu Associate professor

Shanxi University of Traditional Chinese Medicine

69 Wei Ding Associate professor

department of traditional Chinese medicine, the first affiliated hospital with Nanjing Medical University

70 Tian-min Wu

Associate professor

department of traditional Chinese medicine combine traditional Chinese and western medicine, The first hospital affiliated to fujian medical university

71 Xiu-ping Bai

Archiater The second ward, Department of Cardiology,Four affiliated hospital, Harbin medical university

72 Wei-gang Jia


emergency department, Heilongjiang academy of TCM

73 Rong Lv Associate Shanghai University of Chinese Medicine

professor

74 Li-qun Xie

The Deputy Director Dr

the first affiliated hospital with Nanjing Medical University

75 Yong Yang

Associate professor

Shandong Traditional Chinese Medicine University

76 De-biao Chi

Associate professor

college of pharmacy, Southern Medical University

77 Chao-yang Wang

Director Xiamen hospital of T.C.M

78 Yin Long Associate chief physician

department of traditional Chinese medicine , Xiking hospital, fourth military medical university

79 Jiang Chang


Shanxi province hospital of traditional Chinese medicine

80 Guan-hai Dai

Associate

professor

Zhejiang institute of traditional Chinese medicine

81 Chun-yu Zeng

Archiater Department of Cardiology, Third military medical university, third affiliated hospital

82 Tao Zhou Archiater Henan college of traditional Chinese medicine the first affiliated hospital

83 Fan Li Associate professor

Pathological physiology teaching and research section, Kunming Medical University

84 Tao Xu Associate chief physician

Department of Cardiology, The second affiliated hospital of guiyang college of traditional Chinese medicine

85 Jian Wang

Archiater Changchun university of Chinese medicine affiliated hospital

86 Yong-hui Wang

Associate professor

Shanxi University of Traditional Chinese Medicine

87 Yan Xiao Associate chief physician

Urumqi hospital of traditional Chinese medicine

88 Wei Fang Director of TCM doctor

Chinese medicine hospital of hangzhou

89 Xiao-yun Zhang

Professor Hebei North University

90 Guo-qing Zheng

Associate professor

The second hospital affiliated to wenzhou medical college

91 Qian Wang


Cardiovascular Beijing hospital of traditional Chinese medicine, Capital medical university

92 Bao-qi Wang

Associate professor

Henan college of traditional Chinese medicine

93 Xiao-yan Associate Henan college of traditional Chinese medicine

Fang professor 94 Heng-hao

Nie Associate chief physician

Thrombosis combine traditional Chinese and western medicine medical center in liaoning province

95 Ze-jun Huo


department of traditional Chinese medicine , Peking University third hospital

96 Wen Jin Associate professor

The Institute of Medicinal Plant Development

97 Li-xia Zhang


department of nephropathy ,ethune international peace hospital of Beijing military area command

98 Zhe Zhang

Associate professor

Liaoning university of traditional Chinese medicine

Acknowledgments

天士力制药集团股份有限公司

Tasly Pharmaceutical Group Co., Ltd.

北京吉安得尔科技有限公司

Gene&I Scientific Ltd.

瑞典帕瑞医学（中国）公司

PERIMED CHINA LTD.

哈尔滨珍宝岛医药贸易有限公司

Harbin ZBD Medicine&Trade Co.,Ltd.

the 13th annual conference of the professional committee ... material/icm 2013.pdfinternational...

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