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Acta Pharmacologica Sinica 2004年第4期25卷 87-91页

作者： Qing ZHAO, Nan CHEN2, Wei-ming WANG, Jian LU3,Bing-bing DAI3 department of Nephrology, Ruijin Hospital 3department of biochemistry, shanghai second medical university, shanghai 200025, china department of Nephrology Ruijin Hospital

AIM: To investigate the regulatory mechanism of transforming growth factor-betaon activity of connective tissue growth factor promoter in mouse NIH/3T3 fibroblasts. METHODS: The regulation fragment of the 5' flanking region of the human CTGF gene was linked to pGL3-Basic vector, a firefly luciferase reporter construct without promoter. The recombinant plasmid pCTGF-luc was transiently transfected to NIH/3T3 fibroblasts. The activity of CTGF promoter after treatment of TGF-β1 and MAPK pathway inhibitors were assayed with luciferase reporter gene assay system. RESULTS: TGF-β1-induced increase of CTGF promoter activity was concentration-dependent, with a plateau at 5 μg/L by 2.67-fold vs control (P3580 (10 μmol/L), the p38 MAP kinaseinhibitor, decreased basal and TGF-β1-induced activation of CTGF promoter. However, inhibition of c-Jun-N-terminal kinase/stress-activated protein kinase by SP600125 (20 μmol/L) was without effect. CONCLUSION: TGF-β1stimulated the transcriptional activity of CTGF gene promoter in NIH/3T3 fibroblasts in a dose- and time-dependent manner. MAPK pathway may play a role in the regulation of TGF-β1-induced CTGF expression.

关键词： transforming growth factor-beta connective tissue growth factor promoter mitogen-activated protein kinases signal transduction

The convergent point of the endocytic and autophagicpathways in leydig cells

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Cell Research 1999年第4期9卷 243-253页

作者： YI JING XUE MING TANG(department of Biophysics and Cell Biology, shanghai second medical university, shanghai 200025, china) department of Biophysics and Cell Biology Shanghai Second Medical University Shanghai 200025 China

INTRODUCTIONEndocytosisandalltophagocytosis(alltophagy)arethemajorpathwaysformaterialstobetransportedtolysosolnesillanimalcells,theformerbeingresponsibleforuptakeofextracellular***... 详细信息

INTRODUCTIONEndocytosisandalltophagocytosis(alltophagy)arethemajorpathwaysformaterialstobetransportedtolysosolnesillanimalcells,theformerbeingresponsibleforuptakeofextracellular***...

关键词： Autophagocytosis endocytosis endosomes lysosomes.

Expression of Hepatitis C Virus E2 Ectodomain in *** and Its Application in the Detection of Anti-E2 Antibodies in Human Sera

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Acta Biochimica et Biophysica Sinica 2004年第1期36卷 57-63页

作者： Jing LIU, Xin-Xin ZHANG, Shen-Ying ZHANG, Min LU, Yu-Ying KONG, Yuan WANG, and Guang-Di LI ( State Key Laboratory of Molecular Biology, Institute of biochemistry and Cell Biology, shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, shanghai 200031, china 2 department of Infectious Diseases, Ruijin Hospital, second medical university of shanghai, shanghai 200025, china ) State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences Shanghai 200031 Chin

The second envelope glycoprotein （E2） of hepatitis C virus has been shown to bind human target cells and has become a major target for the development of anti-HCV vaccines. Anti-E2 antibodies have been suggested to be of clinical significance in diagnosis, treatment and prognosis of hepatitis C. However, large-scale expression and purification of E2 proteins in mammalian cells is difficult. As an alternative, E2 fragment （aa 385-730） with a four-amino-acid mutation （aa 568-571 PCNI to RVTS） was expressed as hexa-histidine-tagged full length protein [E2N730（m）] in E. coli and purified to over 85% purity. Purified E2N730（m） was specifically recognized by homologous hepatitis C patient serum in Western blot, suggesting that it displayed E2specific antigenicity. Rabbit antiserum raised against E2N730（m） recognized E2 glyco-proteins expressed in mammalian cells in Western blot. Purified E2N730（m） was used to detect anti-E2 antibodies in human sera and showed better specificity and sensitivity than previousl

关键词： hepatitis C virus envelope protein E2 expression and purification Escherichia coll

Changes in Autoantibodies against β1-Adrenoceptor and M2-Muscarinic Receptor during Development of Renovascular Hypertension in Rats

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Acta Biochimica et Biophysica Sinica 2004年第12期36卷 793-797页

作者： Jian-Ming ZHI*, Rong-Fang CHEN1, Heng WU2, Hui-Rong LIU2, and Rong-Rui ZHAO2 department of Physiology, shanghai second medical university, shanghai 200025, china 1department of Toxicology, second Military medical university, shanghai 200433, china 2 department of Physiology, Shanxi medical university, Taiyuan 030001, china department of Physiology Shanghai Second Medical UniversityShanghai 200025 China department of Toxicology Second Military Medical UniversityShanghai 200433 China department of Physiology Shanxi Medical UniversityTaiyuan 030001 China

In an experimental rat’s renovascular hypertension model, we studied the genesis of anti- cardiac β1-adrenoceptor and M2-muscarinic receptor autoantibodies in relation to the changes in immunological function during the development of renal hypertension. The biological activities of these autoantibodies were also examined. It was shown that after two weeks of operation both the frequency of occurrence and the titre of autoantibodies to cardiac β1-adrenoceptor and M2-muscarinic receptor were significantly increased as compared with the control of pre-treatment. The increased autoantibodies lasted for several weeks and then automatically decreased gradually to the pre-clipping level at 10 weeks. Meanwhile the ratio of CD4 /CD8 + + was also undergone an initial increase followed by gradual recovery and correlated well with the changes in antibody titre. The biological effects of these autoantibodies displayed an “agonistic-like” activities on the beating frequency of cultured neonatal cardiomyocyte. It is suggested that autoimmune mechanisms are involved in the pathogenesis of renal hypertension and the cardiac receptor autoantibodies might be one of the mechanisms leading to cardiac dysfunction.

关键词： autoantibody renal hypertension cardiac receptor immunology

Effects of Ginkgolide B on action potential and calcium, potassium current in guinea pig ventricular myocytes

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Acta Pharmacologica Sinica 2004年第2期 77-81页

作者： Xiao-yan QI, Zhi-xiong ZHANG2, You-qiu XU3 department of Physiology, shanghai university of Traditional Chinese medicine, shanghai 200032 3department of Physiology, shanghai second medical university, shanghai 200025, china

AIM: To investigate the effect of Ginkgolide B (GB) on action potential (AP) , delayed rectifier potassium current (IK), and L-type calcium current (ICa-L) in guinea pig ventricular myocytes. METHODS: Single ventricular myocytes were isolated by an enzymatic dissociation method. AP, IK, ICa-L were recorded by whole-cell patch-clamp tech- nique in either current or voltage clamp mode. RESULTS: GB shortened APD in a concentration-dependent manner. GB 0.1, 1, and 10 μmol/L shortened APD50 by 7.9 % (n=5, P>0.05), 18.4 % (n=5, P05), 17.6 % (n=5, P3.1 % (n=6, P37.7 % (n=6, P<0.05), respectively at testing potential of +50 mV and shift the I-V curve of Ik upward. But GB had no significant effect on ICa-L at above concentrations. CONCLUSION: GB significantly shortened APD in a concentration-dependent manner which mainly due to increase of IK.

关键词： Ginkgo biloba patch-clamp techniques myocardium action potentials potassium channels cal- cium channels

About the evaluation of drug combination

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Acta Pharmacologica Sinica 2004年第2期 20-21页

作者： Zheng-jun JIN1department of Pharmacology, shanghai second medical university, shanghai 200025, china

Germ cell apoptosis induced by Ureaplasma urealyticum infection

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Asian Journal of Andrology 2001年第3期3卷 199-204页

作者： Chen XU Mei-Ge LU Jing-Sheng FENG Qang-Su Guo Y-Fei WANG department of Histology & Embryology, shanghai second medical university, shanghai 200025, china department of Histology & Embryology Shanghai Second Medical University Shanghai 200025 China

Aim: To study the effect of Ureaplasma urealyticum (UU) infection on germ cell apoptosis of male rats. Meth-ods: Male rats were infected artificially with UU serotype 8 (T_(960)). Morphological changes of germ cells in the sem-iniferous tubules and the lumen of the epididymides were observed under the light microscope. Fluorescence-conjugatedpolyclonal antibodies to Fas and Fas ligand (FasL) were used to localize Fas and FasL. TUNEL staining of germ cellsand Sertoli cells was performed by the AKPase method. TUNEL-positive rate (% positive cells) and TUNEL-positivearea (area occupied by stained cells) were analysed by KS400 Image Analysis System. The DNA laddering analysiswas performed by agarose gels electrophoresis. Results: In those rats infected with UU; (1) Exfoliated germ cellswere dramatically increased. Many multinucleated giant cells were found in the seminiferous tubules and the lumen ofthe epididymides. (2) The number of TUNEL-positive cells and the TUNEL-positive area were significantly increased.(3) The expression of Fas and FasL in germ cells and Sertoli cells was up-regulated. (4) Discrete bands of fragmentedDNA were found in the testicular cells. Conclusion: In male rats, germ cell apoptosis was increased in UU infec-tion . (Asian J Androl 2001 Sep; 3: 199 - 204)

关键词： germ cells apoptosis Ureaplasma urealyticum Fas/FasL DNA electrophoresis

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Predominant expression of β-chain variable region genes of T-cell receptor in TILs from human renal cell carcinoma

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Chinese Science Bulletin 1997年第8期42卷 693-697页

作者： LI Xueyi, WANG Ying, ZHENG Zhongcheng, ZHANG Dongqing, FUSaili, LING Jianyu and ZHOU Guangyan (CHOU Kuangyen)1. shanghai Institute of Immunology, shanghai second medical university, shanghai 200025, china 2. shanghai Institute of biochemistry, Chinese Academy of Sciences, shanghai 200031, china 3. shanghai Ruijin Hospital, shanghai 200025, china shanghai Institute of Immunology Shanghai Second Medical University Shanghai China shanghai Institute of biochemistry Chinese Academy of Sciences Shanghai China shanghai Ruijin Hospital Shanghai China

THE repertoire of T-cell antigen receptor(TcR)can be effectively analysed by an approachwith quantitative polymerase chain reaction(qPCR),which allows to characterize the ex-panding T-cell clones by the patterns of re... 详细信息

关键词： renal cell carcinoma tumor-infiltrating lymphocyte T-cell receptor Vβ gene quantitative polymerase chain reaction.

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Dicoumarol Alters Cellular Redox State and Inhibits Nuclear Factor Kappa B to Enhance Arsenic Trioxide-Induced Apoptosis

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Acta Biochimica et Biophysica Sinica 2004年第3期36卷 235-242页

作者： Ya-Wu JING, Jing YI, Yu-Ying CHEN, Qing-Shen HU, Gui-Ying SHI, Hue LI, and Xue-Ming TANG* department of Cell Biology, shanghai second medical university, shanghai 200025, china department of Cell Biology Shanghai Second Medical University

The effects of a number of cytotoxic drugs are influenced by cellular reduction/oxidation (redox) state. In the present study, we attempt to explore if dicoumarol, an inhibitor of NADPH: quinone oxidoreductase (NQO1), alters the cellular redox state and how this alteration affects the redox-related apoptosis. Flow cytometry was used to assess the reactive oxygen species (ROS) level and apoptotic rates of HeLa cells treated with arsenic trioxide (As2O3) alone or in combination with natural anthraquinone emodin and dicoumarol or plus N-acetyl-cysteine. Western blot, immunofluorescence, electrophoretic mobility shift assay and luciferase assay were used to detect Nuclear Factor kappa B (NF-κB) activation. The results showed that dicoumarol synergized with emodin to sensitize HeLa cells to As2O3-induced apoptosis through raising the ROS level. More notably, this enhanced susceptibility was associated with a ROS-mediated inhi- bition of NF-κB activation in which the combinative treatment with dicoumarol prevented NF-κB from binding to target DNA. It was suggested that dicoumarol in combination with anthraquinones might be a novel strategy to expand the chemotherapeutic spectrum of As2O3 by means of interfering the cellular redox state.

关键词： dicoumarol arsenic trioxide reduction/ oxidation state apoptosis NF-κB

Upregulation of heparin-binding growth-associated molecule after spinal cord injury in adult rats

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Acta Pharmacologica Sinica 2004年第5期25卷 69-74页

作者： Yan-ting WANG2, Shu HAN2, Kai-hua ZHANG2, Yu JIN3, Xiao-ming XU2,4, Pei-hua LU2,52department of Neurobiology, shanghai second medical university, shanghai 200025, china 3department of Surgery,Xinhua Hospital Affiliated to shanghai second medical university 4Kentucky Spinal Cord Injury Research Center,department of Neurological Surgery, university of Louisville School of Medicine, Louisville, KY, 40292, USA department of Neurobiology Shanghai Second Medical University Shanghai 200025 China

AIM: To investigate whether traumatic spinal cord injury (SCI) induces changes of gene expression of heparin-binding growth-associated molecule (HB-GAM). METHODS: In a spinal cord transection model, HB-GAM ex-pression and cellular localization were examined using Northern blot, RT-PCR, immunohistochemistry and immun-ofluorescence double-labeling methods. RESULTS: HB-GAM mRNA was significantly upregulated in spinal cordtissues rostral and caudal to the injury at 7 d after SCI. HB-GAM gene expression was markedly increased at 3 d,peaked at 7 d, and declined to the baseline level at 28 d post-injury. During its peak expression, HB-GAM was co-localized in astrocytes, oligodendrocytes, and neurons in spinal cord tissues within 7 mm from the site of spinaltransection. CONCLUSION: SCI induces HB-GAM expression at both mRNA and protein levels in areas close tothe injury. Both neurons and glial cells expressed HB-GAM implying that HB-GAM played a role in the process ofinjury and/or repair following SCI.

关键词： heparin-binding growth-associated molecule spinal cord injuries immunohistochemistry reversetranscriptase polymerase chain reaction astrocytes neurons