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Activation of PI3-K/Akt pathway for thermal preconditioning to protect cultured cerebellar granule neurons against low potassium-induced apoptosis

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Acta Pharmacologica Sinica 2007年第2期28卷 173-179页

作者： Lin CAO~(2,3) De-xiong CAO~3 Xing-wen SU~2 Li-jun ChEN~2 Ai-lin LIU~2 Wei-jian JIAN~2 Yu-ping WU~2 Peng-xin QIU~2 Guang mei YAN~(2,4)2 department of Pharmacology,Zhongshan Medical School,sun yat-sen university,guangzhou 510080,china 3 department of Anesthesiology The Second Affiliated hospital,sun yat-sen university,guangzhou 510120,china department of Pharmacology Zhongshan Medical School Sun Yat-sen University Guangzhou China department of Anesthesiology The Second Affiliated Hospital Sun Yat-sen University Guangzhou China

Aim:This study was designed to investigate whether the activation of thephosphatidylinositol 3-kinase(PI3-K)/Akt pathway is required for thermal pre-conditioning to protect rat cerebellar granule neurons(CGN)against apoptosisinduced by low potassium,and to explore the possibility of a link between theupregulated heat shock protein(hSP)70 expression and Akt activation in theacquisition of neuroprotection induced by thermal ***:CGN cultured for 8 d in vitro were switched to 5K medium for 24 h after thermalpreconditioning(TP;43.5 ℃ for 90 min,then 37 ℃ for 1 h).To study the role of thePI3-K/Akt pathway,a PI3-K inhibitor,LY294002(20 μmol/L)was added into thecultures 1 h before TP.3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bro-mide(MTT)assay and fluorescein diacetate staining were used to determine *** 33258 staining and agar gel electrophoresis were used to testthe morphological and biological characters of *** blot analysis wasemployed to detect the levels of phospho-Akt,phospho-glycogen synthase ki-nase 3β(GSK3β) Akt,GSK3β,and ***:TP protected CGN againstapoptosis induced by low ***294002 inhibited the neuroprotectiveeffect on CGN induced by *** induced a robust activation of Akt and theinactivation of GSK3β via ***,the activation of the PI3-K/Aktpathway by TP persisted for 24 h in the 5K ***294002(20 μmol/L)failedto inhibit the upregulated hSP70 expression induced by ***:Theactivation of the PI3-K/Akt pathway is required for TP to protect CGN againstapoptosis induced by low potassium,but the neuroprotective effect by Akt acti-vation is not mediated through the downstream induction of hSP70 expression.

关键词： thermal preconditioning heat cerebellum neurons apoptosis LY294002 phospho Akt phospho-GSK3β hSP70

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Effects of Qingkailing on Experimental Allergic Uveitis in rabbits

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眼科学报 1992年第3期8卷 138-142页

作者： Shixing hu Shaozhen Li Chufang Xie Suhua Pan Xiangkun huang Winifred Mao Zhongshan Ophthalmic center,sun yat-sen university of Medical Sciences guangzhou 510060,china Zhongshan Ophthalmic center Sun Yat-sen University of Medical Sciences Guangzhou China.

Experimental allergic uveitis(EAU)in rabbits was induced by singleintraocular injection of bovine serum albumin(BSA).The average of protein con-centration in aqueous humor of untreated group of rabbits was 14.33±1.21mg/mland the count of tritiated thymidine(~3h-Tdr)incorporated into lymphocyte T was3,987±1,156cpm/10~*** specific antibody responses to BSA in the serum andthe aqueous were 0.508±0.034 and 0.369±0.019(OD)***,the effect of Qingkailing on EAU was observed in comparison wi...

关键词： experimental allergic uveitis (EAU) tritiated thymidine (~3h-Tdr) qingkailing

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BOUNDArY PENALTY FINITE ELEMENT METhODS FOr BLENDING SUrFACES,I BASIC ThEOrY

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Journal of Computational Mathematics 1998年第5期16卷 457-480页

作者： Zi-cai Li (department of Applied Mathematics, National sun yat-sen university,Kaohsiung 80242, Taiwan) National sun yat-sen Univerisy 台湾

When parametric functions are used to blend 3D surfaces, geometric continuity of displacements and derivatives until to the surface boundary must be satisfied. By the traditional blending techniques, however, arbitrariness of the solutions arises to cause a difficulty in choosing a suitable blending surface. hence to explore new blending techniques is necessary to construct good surfaces so as to satisfy engineering requirements. In this paper, a blending surface is described as a flexibly elastic plate both in partial differential equations and in their variational equations, thus to lead to a unique solution in a sense of the minimal global surface curvature. Boundary penalty finite element methods (BP-FEMs) with and without approximate integration are proposed to handle the complicated constraints along the blending boundary. Not only have the optimal convergence rate O(h(2)) of second order generalized derivatives of the solutions in the solution domain been obtained, but also the high convergence rate O(h(4)) of the tangent boundary condition of the solutions can be achieved, where h is the maximal boundary length of rectangular elements used. Moreover, useful guidance in computation is discovered to deal with interpolation and approximation in the boundary penalty integrals. A numerical example is also provided to verify perfectly the main theoretical analysis made. This paper yields a framework of mathematical modelling, numerical techniques and error analysis to the general and complicated blending problems.

关键词： blending surfaces parametric surfaces plate mathematical modelling variational equations finite element methods boundary penalty method computer geometric aided design

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Thermal preconditioning protected cerebellar granule neurons of rats by modulating hSP70 expression

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Acta Pharmacologica Sinica 2004年第4期25卷 60-63页

作者： Li-jun ChEN, Xing-wen SU, Peng-xin QIU, Yi-junhUANG, Guang-mei YAN2 department of Pharmacology, Zhongshan MedicalCollege, sun yat-sen university, guangzhou 510089, china department of Pharmacology Zhongshan Medical College Sun YatSen University Guangzhou 510089

AIM: To explore the possibility that expressions of different heat shock proteins (hSPs) are specifically involved in the protection of thermal preconditioning against apoptosis of cerebellar granule neurons (CGNs) induced by repolarization. METhODS: Western blotting was used to detect expressions of hSP27, hSP70, and hSP90 in- duced by thermal preconditioning (TP) in CGNs; reverse transcription-polymerase chain reaction (rT-PCr) was used to detect the expression level of hSP70 mrNA; apoptosis of CGNs was induced by switching culture medium containing KCl 25 mmol/L to one containing KCl 5 mmol/L. rESULTS: No expression of hSP27 in cerebellar granule neurons was observed with TP at 44 oC. Expression of hSP90 was obvious in CGNs both without and with TP at 44 oC for different periods. Expression of hSP70 protein in CGNs was lower with TP at 44 oC for 5 min, but it increased gradually after the period was prolonged to 30, 60, or 90 min. hSP70 mrNA was detected after TP 44 oC for 30, 60, and 90 min and increased gradually with time. hSP70 antisense oligodeoxynucleotides 10 μmol/Lfor 72 h inhibited the protective effects of TP at 44 oC on apoptosis of CGNs induced by repolarization. CONCLUSION: hSP70 is involved in protective effects of thermal preconditioning on apoptosis in cerebellar granule neurons induced by repolarization.

关键词： thermal preconditioning heat heat shock proteins cerebellum neurons apoptosis antisense oligodeoxynucleotides Western blotting

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Microtubule stability and MAP1B upregulation control neuritogenesis in CAD cells

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Acta Pharmacologica Sinica 2006年第9期27卷 1119-1126页

作者： Wen LI~(2,5)Jin-tang XIA~3 Yue FENG~(4,5)2 Laboratory of Surgery,The First Affiliated hospital,sun yat-sen university,guangzhou 510080,china 3 department of General Surgery,The First People's hospital of guangzhou,guangzhou 510180,china 4 department of Pharmacology,Emory university School of Medicine,USA Laboratory of Surgery The First Affiliated Hospital Sun Yat-sen University Guangzhou China department of General Surgery The First People's Hospital of Guangzhou Guangzhou China department of Pharmacology Emory University School of Medicine USA

Aim:To study the role of microtubule dynamics and microtubule associated pro-tein 1B(MAP1B)in regulation of the neurite extension in CAD catecholaminergicneuronal cell ***:The neuritogenesis of the CAD cells was abolishedby inhibiting microtubule polymerization with nocodazole and by blocking micro-tubule depolymerization with ***1B and tubulin protein expression wasdetected by Western *** staining of tubulins was observedby fluorescent and confocal ***:Microtubule dynamics wasessential for CAD neurite *** analysis revealed that neurite exten-sion was much more sensitive to nocodazole than to taxol,suggesting a func-tional requirement for highly active microtubule assembly.A remarkableupregulation of MAP IB protein was detected during neurite extension accompa-nied with increased microtubule ***:Upregulation of MAP1Bleads to the stabilization of newly formed microtubules in the developing neurites,which in turn promotes neurite extension.

关键词： microtubule stability microtubule associated protein 1B neuritogenesis

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Studies on human Fetal Lens Crystallins Under Oxidative Stress and Protective Effects of Tea Polyphenols

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眼科学报 1998年第3期14卷 170-175页

作者： hui Xiang, Suhua Pan, Shaozhen LiGuangdong Provincial Corps hospital, Chinese People’ s Armed Police Forces, guangzhou 510010, china Zhongshan Ophthalmic center, sun yat-sen university of Medical Sciences, guangzhou 510060, china Guangdong Provincial Corps hospital Chinese People's Armed Police Forces Guangzhou 510010 China

Purpose: To investigate the oxidative modification of water-soluble crystallins of human fetal lens with h2O2 and fourteen metal ions with or without EDTA. Tea-polyphenols (TP) was added to above solutions in order to testing their antioxidative abilities. Methods: The experiments were performed at 37℃ with final concentration of 2.5mg/ml protein, 0.1 mM metal ions, 0.3 mM EDTA and 1.0 mM h2O2. Then the TP was added to the solution with CuSO4 and h2O2, after 5 or 24 hours, the crystallins were analysed with SDS-PAGE and *** : There were marked oxidative modifications of lens protein in h2O2 and copper without EDTA. In SDS-PAGE patterns, we found an increase in those species above of bands higher than 30 kD and some diffuse bands from 30 to 17 kD after 5 hours. In IEF patterns, there were a general increase on acidity with loss of the more basic species. When the TP was added, there was not any difference with control ***: The results indicate that exposure of water-soluble

关键词：人胎儿晶状体晶体蛋白氧化变性抗氧化作用多酚

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Muscarinic receptor Agonists Protect Cultured Bovine Trabecular Meshwork Cells against Apoptosis Induced by Dexamethasone

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眼科学报 2004年第1期20卷 42-47页

作者： Yajuan Gu1,Shujun Zeng2,PengXin Qiu2,Yuping Wu2,Dawei Peng2,Guangmei Yan11department of Pharmacology, Zhongshan Medical College, sun yat-sen university, guangzhou 510089, china2Zhongshan Ophthalmic center, sun yat-sen university, guangzhou 510060, china department of Pharmacology Zhongshan Medical College Sun Yat-sen University Guangzhou 510089 China.

Purpose:To study whether muscarinic receptor agonists can protect cultured bovine trabecular meshwork cells against apoptosis induced by ***:The third to fifth passags of bovine trabecular meshwork cells were grown to confluence and incubated for 1～14 days in growth media with dexamethasone or pretreatment of pilocarpine or *** cultures were evaluated for apoptosis by phase-contrast microscopy, fluorescence microscopy, DNA laddering and flow cytometric ***:Dexamethasone (0.24～0.96 mmol·L-1) induced apoptosis of trabecular mes-hwork cells in a dose and time-dependent *** 0.48 mmol·L-1 dexametha-sone-treatment, 1.84 mmol·L-1 of pilocarpine or 2.74 mmol·L-1 of carbachol added could significantly reduce apoptotic ***:Muscarinic receptor agonists can protect cultured bovine trabecular meshwork cells against apoptosis induced by dexamethasone. Eye Science 2004;20:42-47.

关键词： apoptosis dexamethasone pilocarpine carbachol trabecular meshwork glaucoma

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Clinicopathological analysis of 14 patients with combined hepatocellular carcinoma and cholangiocarcinoma

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hepatobiliary & Pancreatic Diseases International 2011年第6期10卷 620-625页

作者： Xian-huan Yu,Lei-Bo Xu,hong Zeng,rui Zhang,Jie Wang and Chao LiuAuthor Affiliations:department of hepato-Pancreato-Biliary Surgery and department of Pathology,sun yat-sen Memorial hospital,sun yat-sen university,guangzhou 510120,china department of hepato-Pancreato-Biliary Surgery Sun Yat-Sen Memorial Hospital Sun Yat-Sen University Guangzhou 510120 China department of Pathology Sun Yat-Sen Memorial Hospital Sun Yat-Sen University Guangzhou 510120 China

BACKGrOUND:Combined hepatocellular carcinoma and cholangiocarcinoma (chCC-CC) is an uncommon subtype of primary hepatic carcinoma,and its prognosis is *** study was undertaken to investigate the prognosis and the clinicopathological characteristics of chCC-CC,including their possible cellular ***:Among 852 patients with a primary hepatic carcinoma who underwent hepatectomy from January 1998 to April 2008 at our hospital,chCC-CC was identified in 14 patients The clinicopathological characteristics of the 14 patients were analyzed *** expression of the liver stem cell markers (c-kit,CD90,CD133 and CK19) in the tumor tissue was detected by immunohistochemistry,and the Kaplan-Meier method was used to evaluate ***:Among the 14 patients,9 presented with abdominal pain,3 with anorexia and debilitation,and the remaining two patients were *** mean age was 53.6±3.0 (range 38-74) *** the included patients,11 had an elevated serum alpha-fetoprotein level,13 were infected with hepatitis B virus,9 had vascular invasion and 1 had lymph node metastasis The average diameter of the tumors was 9.9±1.1 (range 5.0-16.0) *** median overall survival time was 7.9±1.0 months In addition,the presence of the liver stem cell markers,c-kit CD90,CD133 and CK19 was 71.4%,85.7%,92.9% and 78.6% *** four markers were simultaneously expressed in eight ***:chCC-CC has aggressive characteristics and the prognosis is extremely *** high expression of liver stem cell markers in the tumor tissue suggests that these tumors may derive from liver stem cells.

关键词： liver neoplasms hepatocellular carcinoma cholangiocarcinoma prognosis stem cells

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Functional distinction of rat liver natural killer cells from spleen natural killer cells under normal and acidic conditions in vitro

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hepatobiliary & Pancreatic Diseases International 2012年第3期11卷 285-293页

作者： Li-hong Lv, Jian-Dong Yu, Guo-Lin Li, Tian-Zhu Long, Wei Zhang, Ya-Jin Chen, Jun Min and Yun-Le Wan department of hepatobiliary Surgery, sun yat-sen Memorial hospital, sun yat-sen university department of Breast Surgery, guangzhou Women and Children’s Medical center guangzhou 510120, china department of hepatobiliary and Pancreatic Surgery, First Affiliated hospital, Zhejiang university School of Medicine, hangzhou 310003, china department of hepatobiliary Surgery Sun Yat-Sen Memorial Hospital Sun Yat-Sen University (Lv LH Yu JD Li GL Chen YJ Min J and Wan YL) guangzhou 510120 China department of Breast Surgery Guangzhou Women and Children's Medical Center (Long TZ) department of hepatobiliary and Pancreatic Surgery First Affiliated Hospital Zhejiang University School of Medicine Hangzhou 310003 China

BACKGrOUND: The microenvironment within solid tumors has often been shown to exhibit an acidic extracellular ph. Although the morphologic and functional differences in natural killer (NK) cells of the liver and spleen have been reported previously under physiological conditions, the difference under acidic conditions is still unclear. This study was to investigate the differences in the morphological and functional characteristics between rat liver and spleen NK cells under normal and acidic conditions in vitro. METhODS: Liver and spleen NK cells were isolated and purified from Sprague-Dawley rats by density gradient centrifugation and the Dynabeads FlowComp TM Flexi system, and stimulated for 4 days with or without IL-2 or treated with low ph or control for different times. Morphology was examined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM), cell death and proliferation assays were performed by flow cytometry, IFN-γ production was tested by ELISA, and cytotoxic activity was evaluated by lactate dehydrogenase (LDh) release assay. rESULTS: Liver NK cells had significantly higher levels of cytotoxic activity than spleen NK cells under normal and acidic conditions, and the maximum difference was observed at ph 5.6. Further analysis revealed that the cytotoxic activity of NK cells was correlated with morphology, cell death, proliferative activity and IFN-γ production. By TEM, liver NK cells contained a greater number of electron-dense granules per cell at ph ***, a modest elevation of cell death and reduction of proliferation of liver NK cells occurred within a range of 5.6-7.2. Interestingly, an acidic extracellular ph only marginally, and not significantly, suppressed IFN-γ production by liver NK cells. CONCLUSION: The sharp morphological and functional differences shown by the two types of NK cells in vitro indicate that liver NK cells are unexpectedly resistant to ph shock.

关键词： natural killer cells liver spleen tumor microenvironment extracellular ph

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relationship between apoptosis and invasive and metastatic potential of hepatocellular carcinoma

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hepatobiliary & Pancreatic Diseases International 2002年第4期1卷 574-576页

作者： En-hua Xiao Guo-Dong hu Jin-Qing Li the department of radiology, Second Xiangya hospital, Central South university, Changsha 410011, china department of radiology, Tongji hospital, Tongji Medical university, Wuhan 430030, china and hepatobiliary department, cancer center of sun yat-sen university of Medical Sciences, guangzhou 510060, china department of radiology Second Xiangya Hospital Central South University Changsha 410011 China. xiaogdk@***

Objective: To study relationship between apoptosis and invasive and metastatic potential of hepatocellu- lar carcinoma (hCC). Methods: Apoptotic rate (Ar), proliferative index (PI) and S-phase fraction (SPF) were measured by flow cytometry, and p170, p21 and nucleoside diphosphate kinase (ndpk) by strept avidin-biotin complex immunohistochemical technique in 57 pa- tients with hCC. results: In this group, Ar was 1.77% ±0.19%, SPF 12.55% ±0.68%, and PI 20.91% ±1.12% (r =-0.173). p170, p21 and ndpk positive rates were 61.36%, 68, 18%, 52.27% respectively in patients with a mean Ar of ≤1.77%, and 23.08%, 38.46%, 84.62% respectively in patients with a mean Ar of >1.77% (all Ph positive tumor invasiveness and metastasis, nd- pk (+) was 43.75%, p21 (+) 75.00%, p170 (+) 65.63%, Ar 1.12% ±0. 16%, PI 23.78% ±1.48%, and SPF 13.90 % ±0.99 %. In patients with negative invasiveness and metastasis, however, ndpk (+) was 80.00%, p21 (+) 44.00%, p170 (+) 36.00%, Ar 2,32%±0.52%, PI 18.53% ±0.82% and SPF 11.43% ±0.70%. Conclusion: Apoptosis of hCC is negatively correla- ted with its invasive and metastatic potential or other factors as proliferative activity, p21, p170 and ndpk.

关键词： hepatocellular carcinoma apoptosis metastatic potential flow cytometry immunohistochemistry

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