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Ohmefentanyl stereoisomers induce changes of CREB phosphorylation in hippocampus of mice in conditioned place preference paradigm

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cell Research 2003年第1期13卷 29-34页

作者： CAN GAO, Li WEI CHEN, JIE CHEN, XUE JUN XU, ZHI QIANG CHISecond Department of Pharmacology, shanghai institute of Materia Medica, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200031, chinaResearch Center for biochemistry and Molecular biology, Xuzhou Medical College, Xuzhou, Jiangsu 221002, china Second Department of Pharmacology Shanghai Institute of Materia Medica Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

The present study was designed to determine the changes of phosphorylation of cAMP- response element binding protein (CREB) in hippocampus induced by ohmefentanyl stereoisomers (F9202 and F9204) in conditioned place preference (CPP) paradigm. The results showed that mice receiving F9202 and F9204 displayed obvious CPP. They could all significantly stimulate CREB phosphorylation and maintained for a long time without affecting total CREB protein levels. The effect of F9204 was similar to morphine which effect was more potent and longer than F9202. We also examined the effects of ketamine, a noncompetitive N-mthyl-D-aspartate receptor (NR) antagonist, on morphine-, F9202- and F9204- induced CPP and phosphorylation of CREB in hippocampus. Ketamine could suppress not only the place preference but also the phosphorylation of CREB produced by morphine, F9202 and F9204. These findings suggest that alterations in the phosphorylation of CREB be relevant to opiates signaling and the development of opiates depende

关键词： cAMP-response element binding protein (CREB) ohmefentanyl stereo isomers opiates hippocampus conditioned place preference (CPP).

The associated regulators and signal pathway in rIL-16/CD4 mediated growth regulation in Jurkat cells

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cell Research 2002年第Z2期 363-372页

作者： XIAO MING ZHANG, YONG HUA XULab of Molecular and cellular Oncology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, 320 Yueyang Road, shanghai 200031, china

IL-16 is a ligand and chemotactic factor for CD4+ T cells. IL-16 inhibits the CDS mediated lymphocyte activation and proliferation. The effects of IL-16 on the target cells are dependent on the cell type, the presence of co-activators etc. To understand the regulation function and mechanism of IL-16 on target cells, we used a 130 a.a. recombinant IL-16 to study its effects on the growth of Jurkat T leukemia cells in vitro. We found that the rIL-16 stimulated the proliferation of Jurkat cells at low dose (10-9M), but inhibited the growth of the cells at higher concentration (10-5M). Results showed that 10-5 M of rIL-16 treatment induced an enhanced apoptosis in Jurkat cells. The treatment blocked the expression of FasL, but up-regulated the c-myc and Bid expression in the cells. Pre-treatment of PKC inhibitor or MEK1 inhibitor markedly increased or decreased the rIL-16 induced growth-inhibiting effects on Jurkat cells, respectively. The results suggested that the rIL-16 might be a regulator for the growth

关键词： IL-16 Jurkat cells growth regulation.

Cloning of rat sp56, the homologue of mouse sperm ZP3 receptor-sp56

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cell Research 2003年第2期13卷 121-129页

作者： SS KOIDE The Population Council Center for Biomedical Research 1230 York Avenue New York New York 100021 USA

Mouse sp56 is considered as one of the candidates for mouse zona pellucida 3 (mZP3) receptor. Up to date, its homologue has only been cloned from guinea pig. namely AM67. Based on the cDNA sequence of mouse sp56, we designed a pair of primer to amplify its homologue from rat testis cDNA. Using RT-PCR, two fragments of 743 bp and 938 bp were amplified. The PCR products show very high homology to mouse sp56. However, the 743 bp product completely lacks one of the seven Sushi domains of mouse sp56. Using the 743 bp product as the probe to detect the expression profile of sp56 in rat tissues, Northern blot shows that a -2.0 kb mRNA expresses specifically in testis. Employed the RACE method, two full cDNA sequences of rat sp56 were obtained. A Mr -42 KD band was detected in denatured and non-reducing protein sample of rat testis and sperm with anti-mouse sp56 monoclonal antibody by Western blot method. Rat sp56 was localized on rat sperm head by the indirect immunofluorescence method. Rat sp56 immunoreactivi

关键词： ZP3 receptor sp56 rat testis sperm cloning.

EMT or apoptosis: a decision for TGF-β

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cell Research 2007年第4期17卷 289-290页

作者： Jianguo Song ~1laboratory of Molecular cell biology,institute of biochemistry and cell biology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031,china aboratoryofMolecularcellbiology InstituteofBiochemistryandCellBiologyShanghaiInstitutesforBiologicalSciencesChineseAcademyofSciencesShanghai200031China

Apoptosis and epithelial-to-mesenchymal transition(EMT)are very fundamental physiological *** are both independent and interrelated events in nor-mal development and in maintaining body *** recent years,EMT has emerged as a focus of studiesin cancer research,and increasing data indicate that EMTfunctions as a central step in the invasion and metastasisof some tumor *** the cytoskeleton rearrange-ment and subsequent morphological changes,EMT is oftencharacterized by the dissolution or attenuation of epithelialadhesion and other polarized structures,for example,tightjunctions,and by accordingly the acquisition of migratoryand invasive properties of the cells undergoing *** these changes,EMT can be associated with transcrip-tional or expressional alterations of some epithelial andmesenchymal *** of apoptosis and EMT are linked withvarious pathological processes,such as fibrosis of liver

关键词： a decision for TGF EMT or apoptosis

Identification of differentially expressed genes in human uterine leiomyomas using differential display

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cell Research 2002年第1期12卷 39-45页

作者： BIN LI, MEI SUN, BIN HE, JIN YU, You DUAN ZHANG, YONG LIAN ZHANG, State Key laboratory of Molecular biology, institute of biochemistry and cell biology, shanghai institute for biological Science, chinese academy of sciences, 820, Yue- Yang Road, shanghai, 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Science Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Science Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Science Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 2OB/GYN Hospital Shanghai (First) Medical University 419 Fang-Xie Road Shanghai China 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Science Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institute for Biological Science Chinese Academy of Sciences 320 Yue-Yang Road Shanghai 200031 China

In searching of differentially expressed genes in human uterine leiomyomas, differential display was used with twelve pairs of primers to compare human uterine leiomyomas with matched myometrium. False positives were eliminated by reverse Northern analysis. Positives were confirmed by Northern blot analysis. RESULTS: [1] Four of 69 cDNA fragments (3 up-regulated named L1, L2 and L3 and 1 down-regulated named Mi in leiomyoma) were confirmed by Northern analysis. [2] Sequence comparison and Northern analysis proved that Li is exactly the human ribosomal protein Si9. [3] It was present ubiquitously in i3 tissues tested but in various levels and even in different size. [4] Li was highly expressed in parotidean cystadenocarcinoma, pancreatic cancer and breast cancer examined. [5] No mutations have been found in human uterine leiomyomas (n=6). CONCLUSIONS: hRPSi9 overexpression might be a universal signal in rapid cell growth tissues.

关键词： Human uterine leiomyomas differentially expressed gene differential display (DD) human S19 ribosomal protein (hRPS19) up-regulated gene in human leiomyomas.

Antitumor Effect of a Novel Adeno-associated Virus Vector Targeting to Telomerase Activity in Tumor cells

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Acta Biochimica et Biophysica Sinica 2004年第7期36卷 492-500页

作者： Yi-Gang WANG1,2#, Jin-Hui WANG1#, Yan-Hong ZHANG1, Qing GU2*, and Xin-Yuan LIU1* 1institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200031, china School of Life Science, Nanchang University, Nanchang 330047, china institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences Shanghai 200031 China

Telomerase activity is a wide tumor marker. Human telomerase reverse transcriptase (hTERT), the catalytic subunit of the telomerase, is transcriptionally upregulated exclusively in about 90% of cancer cells. In this study, we constructed a novel adeno-associated virus (AAV) vector containing the human interferon-β (hIFN-β) gene under the control of hTERT promoter (AAV-hTERT-hIFN-β) and investigated its antitumor effect against various human cancer cells in vitro. AAV-hTERT-hIFN-β displayed cancer-specific hIFN-β expression and cytotoxicity. The cytotoxic ratio was positively correlated with the time length of infection. AAV-hTERT-hIFN-β-mediated apoptotic morphology was observed by transmission electron microscopy. Flow cytometry assay also revealed that the cytotoxicity of AAV-hTERT-hIFN-β was mainly an apoptotic process. These data indicate that AAV in combination with hTERT-mediated therapeutic gene expression may open new possibilities for long-lasting and targeting gene therapy of varieties of cancers.

关键词： adeno-associated virus (AAV) telomerase hTERT promoter human interferon-β cancer

Expression, Purification and Sublocalization of SARS-CoV Nucleocapsid Protein in Insect cells

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Acta Biochimica et Biophysica Sinica 2004年第11期36卷 754-758页

作者： Ai-Xia REN, You-Hua XIE, Yu-Ying KONG, Guan-Zhen YANG, Yao-Zhou ZHANG,Yuan WANG, and Xiang-Fu WUinstitute of biochemistry and Biotechnology, College of Life Science, Zhejiang University, Hangzhou 310029, china State Key laboratory of Molecular biology, shanghai institute of biochemistry and cell biology, shanghai institutes for biological sciences,chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200031, china institute of biochemistry, Zhejiang University of sciences, Hangzhou 310018, china institute of biochemistry and Biotechnology College of Life Science Zhejiang University Hangzhou 310029 China.

The causative agent of severe acute respiratory syndrome (SARS) is a previously unidentified coronavirus, SARS-CoV. The nucleocapsid (N) protein of SARS-CoV is a major viral protein recognized by acute and early convalescent sera from SARS patients. To facilitate the studies on the function and structure of the N protein, this report describe the expression and purification of recombinant SARS-CoV N protein using the baculovirus

关键词： severe acute respiratory syndrome (SARS) SARS coronavirus nucleocapsid protein baculovirus insect cells

Identification and Characterization of a Rat Novel Gene RSEP4 Expressed Specifically in Central Nervous System

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Acta Biochimica et Biophysica Sinica 2004年第7期36卷 501-507页

作者： Xi-Dao WANG1,2, Ling-Wei KONG1, Zhi-Qin XIE1, Yu-Qiu ZHANG3, Zhi-Xin LIN2, Zhi-Qi ZHAO3,Lei YU4, and Nai-He JING1* 1 laboratory of Molecular cell biology, laboratory of Stem cell biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, Graduate School of the chinese academy of sciences, shanghai 200031, china School of Life Science and Biotechnology, 2 shanghai Jiaotong University, shanghai 200030, china institute of Neurobiology, Fudan University, shanghai 200433, china 3 4 Department of cell biology, Neurobiology and Anatomy, College of Medicine, University of Cincinnati, Cincinnati, Ohio, USA laboratory of Molecular cell biology and Stem cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Graduate School of the Chinese Academy of Sciences Shanghai China

The low-abundantly expressed genes composed the majorities of the mRNAs expressed in the central nervous system (CNS), and were thought to be important for the normal brain functions. Through differential screening a low-abundance cDNA sublibrary with mRNA from neuropathic pain of chronic constriction injury (CCI) model, we have identified a novel rat gene, rat spinal-cord expression protein 4 gene (RSEP4). The total length of RSEP4 cDNA is 2006 bp, with a 501 nucleotide open reading frame (ORF) that encodes a 167 amino acid polypeptide. Northern blot revealed that RSEP4 was expressed specifically in the CNS. In situ hybridization showed that the mRNA of RSEP4 was strongly expressed in the CA1, CA2, CA3 and DG regions of hippocampus, the Purkinje cells of cerebellum, and the small sensory neurons of dorsal horn and large motor neurons of ventral horn of spinal cord. Over-expression of RSEP4-EGFP fusion protein in the human embryonic kidney 293T cells showed that RSEP4 protein was mainly localized in the cell cytoplasm. These results suggest that RSEP4 may play some roles in the CNS.

关键词： central nervous system (CNS) rat Northern blot in situ hybridization

Get effective polyclonal antisera in one month

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cell Research 2002年第2期12卷 157-160页

作者： YUAN XIN HU, Ju YUAN QUO, Lu SHEN, YAN CHEN, Zu CHUAN ZHANG, YONG LIAN ZHANGState Key laboratory of Molecular biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, 320 Yue Yang Road, 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China 1State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue Yang Road Shanghai 200031 China

According to the traditional immunization procedure, after the first injection of the sample A (emulsion of aimed antigen and Freund’s complete adjuvant) to immunize rabbit, successive injections of the sample B (emulsion of aimed antigen and Freund’s incomplete adjuvant) were followed every 2-4 weeks. In general, high titer of the corresponding polyclonal antisera will be observed after 4-5 injections of sample B in 3-4 months. This report presents a simply modified procedure that was able to stimulate the antisera formation in one month and achieve enough avidity to satisfy either Western blot or immunohistochemistry analysis. It just applied an additional injection of the sample A to the rabbit at the 3rd day after the primary immunization injection. You could gain the high titer of the antisera right after the first sample B injection in one month. This method has produced the desired results in three different recombinant antigens with different molecular weight (5.9 KD-55 KD) expressed from prokary

关键词： Polyclonal antisera preparation Modified immunization method.