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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

Phage display selection on whole cells yields a small peptide specific for HCV receptor human CD81

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cell Research 2003年第6期13卷 473-479页

作者： JIE CAO, PING ZHAO, XIAO HUI MIAO2, LAN JUAN ZHAO, LI JUN XUE, ZHONG TIAN QIdepartment of Microbiology, second military medical university, shanghai 200433, chinaE-mail: qizt@*** department of Infectious Diseases, Changzheng Hospital, second military medical university, shanghai200003, china department of Microbiology Second Military Medical University Shanghai 200433 China.

The human CD81 (hCD81), the most recently proposed receptor of hepatitis C virus (HCV), can especifically bind to HCV envelope glycoprotein 2 (E2). In this study, hCD81-expressing murine NIH/3T3 cells were used to select hCD81-binding peptides from a phage displayed nonapeptide library (PVIII9aaCys). Eighteen of the 75 clones selected from the library showed specific binding to the hCD/81-expressing NIH/3T3 cells by enzyme linked immunosorbent assay (ELISA) and competitive inhibition test. Twelve out of the 18 clones shared the amino acid motif SPQYWTGPA. Sequence comparison of the motif showed no amino acid homology with the native HCV E2. The motif-containing phages could competitively inhibit the ability of HCV E2 binding to native hCD81-expressing MOLT-4 cells, and induce HCV E2 specific immune response in vivo. These results suggest that the selected motif SPQYWTGPA should be a mimotope of HCV E2 to bind to hCD81 molecules. Our findings cast new light on developing HCV receptor antagonists.

关键词： viral receptor hepatitis C virus cell-based selection hCD81-binding peptide phage display.

The cell cycle related apoptotic susceptibility to arsenic trioxide is associated with the level of reactive oxygen species

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cell Research 2004年第1期14卷 81-85页

作者： Fei GAO~1, Jing YI~1, Jing Qi YUAN~2, Gui Ying SHI~1, Xue Ming TANG~(1,*) ~1department of cell biology, shanghai second medical university, shanghai 200025, china ~2department of Automation, shanghai Jiaotong university, shanghai 200031, china. Tang XM Shanghai Med Univ 2 Dept Cell Biol Shanghai 200025 Peoples R China

Double staining flow cytometry was performed using 7-amino actinomycin D and 6-carboxy-2’,7’-dichlorodihydrofluorescein diacetate, to detect the level fluctuation of reactive oxygen species （ROS） during the cell cycle of normal NB4 cells. Our results showed that NB4 cells possessed higher level of ROS in G2/M phase than in G1 and S phases. Double staining flow cytometry, with TdT mediated dUTP nick end labeling （Tunel） and propidium iodide （PI）, indicated that As2O3 （2μM） could induce apoptosis in NB4 cells prevailingly from G2/M phase, and this efficacy was enhanced upon co-administration of 2, 3-dimethoxy-1, 4-naphthoquinone （DMNQ）（2.5μM） which could produce the endogenous ROS. These results suggested that different ROS level in different cell cycle phases of NB4 cells might determin the selective induction of G2/M apoptosis and the cells’ susceptibility to apoptosis by As2O3.

关键词： arsenic trioxide apoptosis cell cycle reactive oxygen species (ROS).

Effects of rat urotensin Ⅱ on coronary flow and myocardial eNOS protein expression in isolated rat heart

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Acta Pharmacologica Sinica 2004年第11期 60-65页

作者： Ling LI, Wen-jun YUAN, Ding-feng SUdepartment of Pharmacology, department of Physiology, second military medical university, shanghai 200433, china

AIM: To examine the effects of urotensin Ⅱ, a recently discovered endogenous peptide, on coronary flow (CF), cardiac function, and endothelial nitric oxide synthase (eNOS) expression in isolated rat hearts. METHODS: Heart was isolated and perfused retrogradely via the aorta in Langendorff mode. Rat urotensin Ⅱ was administered in the perfusion solution. The eNOS content in myocardium was determined by Western blot. RESULTS: Rat urotensin Ⅱ had no effect on the heart rate, left ventricular systolic pressure, left ventricular end-diastolic pressure, or ±dp/dtmax. While rat urotensin Ⅱ dose-dependently increased CF. CF was increased by 11.43 %, 6.67 %, 6.62 %, 6.56 %, 6.36 %, and 5.86 % respectively in a time-dependent manner at 5, 10, 15, 20, 25, and 30 min after injection of rat urotensin II 6.66×10-2μg. The maximal effect on CF was found at 5 min following urotensin II administration. NG-nitro-L-arginine methyl ester (L-NAME) did not prevent the increased CF in response to urotensin II. Rat uroten

关键词： urotensin Ⅱ nitric oxide synthase nitric oxide heart function tests coronary circulation rats

Effects of esculentoside A on production of interleukin-1, 2, and prostaglandin E2

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Acta Pharmacologica Sinica 2004年第6期25卷 115-119页

作者： Zhen-yu XIAO, Qin-yue ZHENG, Yuan-ying JIANG, Bin ZHOU, Ming YIN2, Hong-bin WANG, Jun-ping ZHANG3 department of Pharmacology,second military medical university, shanghai200433 2College of Pharmacy,Jiao Tong university, shanghai 200030, china department of Pharmacology Second Military Medical University Shanghai 200433 China

AIM: To investigate the influence of esculentoside A (EsA) on immunological function and its mechanism of anti- inflammation. METHODS: Interleukin-1 production was measured by thymocyte co-stimulating assay; the radio- activity of [3H]arachidonic acid (AA) was used to evaluate the release of AA; prostaglandin E2 production was measured with radioimmunoassay (RIA); IL-2 and IFN-γ were detected by ELISA method. RESULTS: EsA (3-12 μmol/L)could potently inhibit the production of IL-1 and PGE2 from both silent and LPS induced macrophages. EsA had no significant effect on the release of AA from murine macrophages. EsA could inhibit the production of IL-2 from murine lymphocytes induced by ConA, but not affect the production from silent lymphocytes. EsA showed no effect on the production of IFN-γ from both silent and ConA induced lymphocytes. CONCLUSION: EsA could affect the immunological function through inhibiting the production of IL-2 from activated splenocytes and the inhibition of production of IL-1 and PGE2 might be one of the anti-inflammation mechanisms of EsA.

关键词： esculentoside A interleukin-1 interleukin-2 prostaglandins E interferon-gamma

Apoptosis in oncology

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cell Research 2001年第1期11卷 1-7页

作者： FAN XIAO QIANG, YA JUN GUO (International Joint Cancer Institute, The second military medical university, shanghai 200423, china) 1International Joint Cancer Institute The Second Military Medical University Shanghai 200433 China 1International Joint Cancer Institute The Second Military Medical University Shanghai 200433 China

Apoptosis is a complex process involving a large array of genes and mutation of any of these genes may lead to malignancy formation. Re-acquirement of FasL by tumor cells may enable them to evade the surveillance of immune system and thus contributes to the growth of tumor. Apart from traditional therapies, inducing apoptosis of tumor cell by new methods employing death receptor ligands and making use of Fas counterattack is also being developed.

关键词： Apoptosis mitochondria death receptor death receptor ligand tumor therapy.

Effect of α-melanocyte- stimulating hormone on proinflammatory cytokines production in neonatal rat astrocytes stimulated with lipopolysaccharide in vitro

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Acta Pharmacologica Sinica 1999年第5期 96-97页

作者： WU Xiu-Ju, TIAN Ye-Ping ( department of Immunology, the second military medical university, shanghai 200433 , china )

Macrophage migration inhibitory factor (MIF) is an

关键词： Effect of melanocyte stimulating hormone on proinflammatory cytokines production in neonatal rat astrocytes stimulated with lipopolysaccharide in vitro MSH

Construction,Expression,and Characterization of a Recombinant Annexin B1-Low Molecular Weight Urokinase Chimera in Escherichia coli

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Acta Biochimica et Biophysica Sinica 2004年第3期36卷 184-190页

作者： Hong-Li YAN, Wei-Ting WANG1, Yan HE , Zhuan-You ZHAO1,Yuan-Jian GAO, Yi ZHANG, and Shu-Han SUN department of medical Genetics, second military medical university, shanghai 200433, china 1department of Pharmacology, Tianjin Institute of Medicine, Tianjin 300193, china department of medical Genetics Second Military Medical University department of Pharmacology Tianjin Institute of Medicine

To produce a thrombi-targeting plasminogen activator, low molecular weight single-chain urokinase gene (scuPA32k) was spliced with the full-length cDNA of annexin B1 gene (anxB1) by overlap extension method. The fused gene anxB1scuPA was ligated into pET28a vector, transformed into E. coli BL21-RIL, and then induced to express under the control of T7 promoter. The AnxB1ScuPA protein ex- pressed amounted to 22% of the total bacterial proteins. The product was refolded, and then purified by using DEAE Sepharose fast flow ion-exchange column and Superdex S-200 gel-filtration column. HPLC analysis revealed that the final purity is about 95%. The specific activity of AnxB1ScuPA, measured as amidolytic activity, reached 100,000 IU/mg. It had a similar S2444 catalytic efficiency (kcat/Km) to ScuPA32k, and also showed high activated-platelet membrane-binding activity and anticoagulant activity, indicating that the chi- mera fully retained the components of enzymatic and membrane-binding activities of the parent molecules. In vivo test revealed that, the dogs administered with AnxB1ScuPA had less reperfusion time, higher reperfu- sion ratio, and less bleeding effects than those with urokinase. These findings indicated that AnxB1ScuPA might have advantages over current available thrombolytic agents.

关键词： annexin B1 thrombolytic agent low molecular weight single-chain urokinase chimera

Clinical analysis of uncinate process carcinoma of the pancreas

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Hepatobiliary & Pancreatic Diseases International 2003年第4期2卷 605-608页

作者： Chun Ye Peng-Cheng xi Xian-Gui Hu the department of General Surgery, Changhai Hospital, second military medical university, shanghai 200433, china department of General Surgery Changhai Hospital Second Military Medical University Shanghai 200433 China. thomas2000@***

OBJECTIVE: To analyse the clinical features of uncinate process carcinoma of the pancreas and the diagnosis and treatment of this malignancy. METHOD: Fifty-nine patients with pancreas uncinate process carcinoma treated from January 1998 to September 2002 at our hospital were analysed retrospectively. RESULTS: Major symptoms of these patients were upper abdominal pain accompanied with lumbar pain, body weight loss and jaundice. Thirty-seven patients received regional pancreaticoduodenectomy (RP), 16 partial resection of the superior mesenteric vein-portal vein (SMV-PV) or superior mesenteric artery (SMA) and reconstruction, 1 anhydrous alcohol injection in the celiac nerve plexus, regional chemotherapy via a chemotherapy pump, and liver biopsy, and 5 no operation. The survival of the patients after operation was 2-46 months (median 12.1 months). Eleven patients are still alive with a longest survival of 46 months. The 1- and 3-year survival rates were 37.7% and 5.6%. CONCLUSIONS: Pancreas uncinate process carcinoma invading the adjacent SMV/SMA-PV causes difficulty in early diagnosis and poor prognosis, which are related to its location, not tumor's aggressive nature. This carcinoma has a high resection rate of 89.8%.

关键词： uncinate process carcinoma pancreas pancreaticoduodenectomy regional

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Corticosterone activates Erk1/2 mitogen-activated protein kinase in primary hippocampal cells through rapid nongenomic mechanism

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Progress in Natural Science:Materials International 2005年第4期 38-43页

作者： QI Aiqun 1, QIU Jian 1, XIAO Lin 2 and CHEN Yizhang 2,3 *(1. department of Physiology, second military medical university, shanghai 200433, china 2. department of Neurobiology, second military medical university, shanghai 200433, china 3. Institute of Neuroscience, second military medical university, shanghai 200433, china) department of Physiology，second military medical university，shanghai 200433，china department of Neurobiology，second military medical university，shanghai 200433，china department of Neurobiology，second military medical university，shanghai 200433，china Institute of Neuroscience，second military medical university，shanghai 200433，china

Nongenomic effects of glucocorticoids (GC) in various cell types have been well documented, but it still remains unknown whether the mechanism also works in hippocampus which is a crucial target of glucocorticoids in neural system during physiological and/or pathophysiological processes. We present here that corticosterone (B) could rapidly activate Erk1/2 mitogen activated protein kinase (MAPK) in primarily cultured hippocampal cells within minutes, with a bell shaped time dependent curve which peaked at 15min and then went down to normal level in 30 min. This activation was blocked by protein kinase C (PKC) inhibitor (G6976), G protein inhibitor (GDPs), and MEK(MAPK/extracellular signal regulated kinase kinase) inhibitor(PD98059), but not by protein kinase A (PKA) inbibitor (H89), tyrosine kinase inhibitor (genistein), and glucocorticoid receptor (GR) antagonist (RU38486). Thus, the rapid activation of Erk1/2 MAPK in primary hippocampal cells induced by B was likely mediated by a G protein coupled receptor (GPCR) pathway with involvement of PKC, which belonged to the nongenomic rather than genomic mechanism of GC's effects.

关键词： glucocorticoid MAPK rapid effects nongenomic mechanism.