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Acta Pharmacologica Sinica 2003年第12期 123-126页

作者： geng jian-guo institute of biochemistry and cell biology, shanghai institutes for biological sciences chinese academy of sciences, shanghai 200031, china

Adhesion and migration of mammalian cells are of crucial importance in a number of biological events, such as fertilization, embryogenesis, pattern, tissue and organ formation, and in a variety of physiological and pathological processes, including lymphocyte trafficking, leukocyte recruitment, hemostasis, wound healing, tumor angiogenesis and cancer metastasis. All these

关键词： of et as in Interaction of vascular endothelial cells with leukocytes thrombosis and cancer growth and metastasis with

Directional migration of leukocytes: their pathological roles in inflammation and strategies for development of anti-inflammatory therapies

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cell Research 2001年第2期11卷 85-88页

作者： geng jian guo institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, 320 Yue Yang Road, shanghai 200031, china institute of biochemistry and cell biology Chinese Academy of Sciences Shanghai. jggeng@***

Directional migration of leukocytes is indispensable to innate immunity for host defense. However, recruitment of leukocytes to a site of tissue injury the constitutes a leading cause for inflammatory responses. Mechanistically, it involves a cascade of cellular events precisely regulated by temporal and spatial present tation of a repertoire of molecules in the migrating leukocytes and their surroundings (microenvironments). Here I will summarize the emerging evidence that has shed lights on the underlying molecular mechanism for directional migration of leukocytes, which has guided the therapeutical development for innovative anti-inflammatory medicines.

关键词： Directional cell migration leukocytes inflammation therapy.

Adenovirus-mediated expression of pig α(1,3) galactosyltransferase reconstructs Gal α(1,3) Gal epitope on the surface of human tumor cells

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cell Research 2001年第2期11卷 116-124页

作者： XING LI, guo HONG XIA, jian FEI, FANG HUANG, LI HE guo institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200031, china 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

Gal α(1, 3) Gal (gal epitope) is a carbohydrate epitope and synthesized in large amount by α(1, 3) galactosyltransferase [α(1, 3) GT] enzyme on the cells of lower mammalian animals such as pigs and mice. Human has no gal epitope due to the inactivation of α(1, 3) GT gene but produces a large amount of antibodies (anti-Gal) which recognize Gal α(1, 3) Gal structures specifically. In this study, a replication deficient recombinant adenoviral vector Ad5sGT containing pig α(1, 3) GT cDNA was constructed and characterized. Adenoviral vector-mediated transfer of pig α(1, 3) GT gene into human tumor cells such as malignant melanoma A375, stomach cancer SGC-7901, and lung cancer SPC-A-1 was reported for the first time. Results showed that Gal epitope did not increase the sensitivity of human tumor cells to human complement-mediated lysis, although human complement activation and the binding of human IgG and IgM natural antibodies to human tumor cells were enhanced significantly after Ad5sGT transduction. Appearance of gal epitope on the human tumor cells changed the expression of cell surface carbohydrates reacting with Ulex europaeus I (UEA I) lectins, Vicia villosa agglutinin (VVA), Arachis hypogaea agglutinin (PNA), and Glycine max agglutinin (SBA) to different degrees. In addition, no effect of gal epitope on the growth in vitro of human tumor cells was observed in MTT assay.

关键词： Adenoviral vector galactosyltransferase Gal α(1 3) Gal gene expression human tumor cell.

Introduction of Foreign Genes into Silkworm Eggs by Electroporation and Its Application in Transgenic Vector Test

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Acta Biochimica et Biophysica Sinica 2004年第5期36卷 323-330页

作者： Xiu-Yang guo#, Liang DONG#, Sheng-Peng WANG, Ting-Qing guo, jian-Yang WANG, and Chang-De LU* institute of biochemistry and cell biology, shanghai institutes for biological sciences, the chinese academy of sciences, 320 Yueyang Road, shanghai 200031, china institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences

Electroporation as a methodology to introduce foreign genes into silkworm eggs was system- atically analyzed. The foreign gene in both the newly hatched and 3rd instar larva DNA can be detected by PCR. The amount of foreign gene in 3rd instar larva DNA was about 1/1000 of that in newly hatched larva DNA. The ratio of foreign gene entering into silkworm eggs was voltage dependent and showed significant difference between the tested silkworm strains. When the piggyBac transposon system was applied, the effect of nuclear localization signal (NLS) peptide and the in vitro transcribed transposase mRNA on the transposi- tion rate has been measured. Results showed that the in vitro transcribed transposase mRNA facilitated trans- position to take place earlier and NLS could result in higher transposition probability and earlier transposition as well. When linearized vectors containing varied length of flanking homologous sequences around a re- porter gene were introduced into silkworm eggs by electroporation, the one with 2.6 kb total arm length gave higher G1 positive ratio than that with total arm length of 1.5 kb and 800 bp.

关键词： silkworm eggs gene introduction electroporation transposon homologous sequences

Overexpression of γ-aminobutyric acid transporter subtype I leads to susceptibility to Kainic acid-induced seizure in transgenic mice

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cell Research 2001年第1期11卷 61-67页

作者： MA YINGHUA, JIA HUA HU, WEN JUAN ZHAO, jian FEI, YUN YU, XIAO GANG ZHOU, ZHEN TONG MEI, LI HE guo (institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200031, china) (shanghai Inst 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China 2shanghai institute of Physiology Chinese Academy of Sciences Shanghai 200031 China 1institute of biochemistry and cell biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China

γ-aminobutyric acid (GABA) is the principal inhibitory neurotransmitter, and the GABAergic synaptic transmission is normally terminated by the rapid uptake through GABA transporters. With transgenic mice ubiquitously overexpressing GABA transporter subtype I (GAT1), the present study explored the pathophysiological role of GAT1 in epileptogenesis. Though displaying no spontaneous seizure activity, these mice exhibit altered electroencephalographic patterns and increased susceptibility to seizure induced by kainic acid. In addition, the GABAA receptor and glutamate transporters are up-regulated in transgenic mice, which perhaps reflects a compensatory or corrective change to the elevated level of GAT1. These preliminary findings support the hypothesis that excitatory and inhibitory neurotransmission, and seizure susceptibility can be altered by neurotransmitter transporters.

关键词： γ-aminobutyric acid transporter seizure susceptibility kainic acid electroencephalography transgenic mice

Transgenic mice overexpressingγ-aminobutyric acid transporter subtypeⅠ develop obesity

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cell Research 2000年第4期10卷 303-310页

作者： MA YING HUA JIA HUA HU XIAO GANG ZHOU RUO WANG ZENG ZHEN TONG MEI jian FEI LI HE guo institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of Science, shanghai 200031, china shanghai institute of harmacy.ankara.edu.tr

Transgenic mice ubiquitously overexpressing murine γ aminobutyric acid transporter subtype Ⅰ were created. Unexpectedly, these mice markedly exhibited heritable obesity, which features significantly increased body weight and fat deposition. Behavioral examination revealed that transgeinc mice have slightly reduced spontaneous locomotive capacity and altered feeding pattern. Tills preliminary finding indicates that the inappropriate level of γ-aminobutyric acid transporters may be directly or indirectly involved in the pathogenic mechanism underlying certain types of obesity.

关键词： γ-aminobutyric acid transporter subtype Ⅰ transgenic mouse obesit€

GABA transporter 1 transcriptional starting site exhibiting tissue specific difference

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cell Research 2001年第2期11卷 161-163页

作者： JIN XIAO PING, FANG HUANG, NA YANG, BU FENG LU, jian FEI, LI HE guo institute of biochemistry and cell biology, shanghai institutes for biological Science, chinese academy of sciences, 320 Yueyang Road, shanghai 200031, china institute of biochemistry and cell biology Chinese Academy of Sciences Shanghai.

GABA transporter 1(GAT1) takes important roles in multiple physiological processes through the uptake and release of GABA, but the regulation of GAT1 gene expression in different tissues is rarely known. To address the question, first, 5’ Rapid amplification of cDNA end (RACE) was used to determine GAT1 transcriptional starting sites in neonatal mouse cerebral cortex and intestine, adult mouse brain and adult rat testis. The products of 5’RACE were confirmed by DNA sequencing. We found that the transcript of GAT1 in neonatal mouse cerebral cortex and adult mouse brain starts at the same site (inside of exon 1), while in mouse intestine, GAT1 starts transcription in intron 1, and in rat testis, the transcript of GAT1 has an additional untranslation exon to the 5’ direction.

关键词： GABA transporter 1(GAT1) transcriptional starting site 5’ RACE tissue spedcific difference DNA sequencing.

Enhancement of germ cell apoptosis induced by ethanol in transgenic mice overexpressing Fas Ligand

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cell Research 2003年第5期13卷 361-367页

作者： JIA HUA HU, JIE jianG, YING HUA MA, NA YANG, MAO HU ZHANG, MIN WU, jian FEI, LI HE guo 1 Laboratory of Molecular cell biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, shanghai 200031, china 2 College of life science, Zhejiang University, Hangzhou 310027, china 3 shanghai Research Center for Biomodel Organism, shanghai 201203, china 1Laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. E-mail: mhzhang@*** 2College of life science Zhejiang University Hangzhou 310027 China 1Laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. E-mail: mhzhang@*** 1Laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. E-mail: mhzhang@*** 1Laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. E-mail: mhzhang@*** 2College of life science Zhejiang University Hangzhou 310027 China 3shanghai Research Center for Biomodel Organism Shanghai 201203 China 1Laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China. E-mail: mhzhang@***

It was suggested that chronic ethanol exposure could result in testicular germ cell apoptosis, but the mechanism is still unclear. In the present study, we use a model of transgenic mice ubiquitously overexpressing human FasL to investigate whether Fas ligand plays a role in ethanol-induced testicular germ cell apoptosis. Both wild-type (WT) mice and transgenic (TG) mice were treated with acute ethanol (20％ v/v) by introperitoneal injection for five times. After ethanol injection, WT mice displayed up-regulation of Fas ligand in the testes, which was shown by FITC-conjugated flow cytometry and western blotting. Moreover, TG mice exhibited significantly more apoptotic germ cells than WT mice did after ethanol injection, which was demonstrated by DNA fragmentation, PI staining flow cytometry and TUNEL staining. In addition, histopathological examination revealed that degenerative changes of epithelial component of the tubules occurred in FasL overexpressing transgenic mice while testicular morphology was

关键词： Fas ligand ethanol apoptosis testes transgenic mouse.

The involvement of p38 MAPK in transforming growth factor β1-induced apoptosis in murine hepatocytes

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cell Research 2001年第2期11卷 89-94页

作者： LIAO JIN HUI, JUN SONG CHEN, MIN QIANG CHAI, SHENG ZHAO, jian guo SONG State Key Laboratory of Molecular biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of sciences, 320 Yue Yang Road, Shan 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

We reported in this manuscript that TGF-β1 induces apoptosis in AML12 murine hepatocytes, which is associated with the activation of p38 MAPK signaling pathway. SB202190, a specific inhibitor of p38 MAPK, strongly inhibited the TGF-β1-induced apoptosis and PAI-1 promoter activity. Treatment of cells with TGF-β1 activates p38. Furthermore, over-expression of dominant negative mutant p38 also reduced the TGF-β1-induced apoptosis. The data indicate that the activation of p38 is involved in TGF-β1-mediated gene expression and apoptosis.

关键词： Transforming growth factor β apoptosis p38 hepatocyte signal transduction.

Identification of glutamate transporters and receptors in mouse testis

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Acta Pharmacologica Sinica 2004年第03期25卷 112-117页

作者： Jia-hua HU,Na YANG2,Ying-hua MA,Jie jianG,Jin-fu ZHANG3,jian FEI4,Li-he guo4 Laboratory of Molecular cell biology,institute of biochemistry and cell biology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031 3Department of Urology,Tongji Hospital,Tongji University,shanghai 200065,china Laboratory of Molecular cell biology Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological Sciences Chinese Academy of Sciences Department of Urology Tongji Hospital Tongji University

AIM: To investigate the presence of glutamate transporters and receptors in mouse testis. METHODS: Glutamate uptake analysis was performed to study the function of glutamate transporters in mouse testis. Comparative RT-PCR technique and sequencing analysis were used to study the expression of glutamate receptors and transport- ers in mouse testis. RESULTS: Mouse testis possessed glutamate uptake capacity with sodium-dependence. Vmax value of glutamate uptake was (1.60 ± 0.21) pmol/min per mg protein and Km value of glutamate uptake was (11.0±1.6) μmol/L in mouse testis according to saturation analysis. Furthermore, the uptake activity could be inhibited by DHK (GLT1 selective inhibitor) and THA (glutamate uptake inhibitor). In addition, RT-PCR results revealed that glutamate transporters (GLT1 and EAAC1) and ionotropic glutamate receptors (NR1, NR2B, GluR6 and KA2) were expressed in mouse testis. CONCLUSION: Glutamate transporters and receptors do exist in mouse testis.

关键词： glutamates receptors amino acid transport systems testis uptake