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  • Cell Research
    Institute of Sports Medicine The Third Hospital Peking University Beijing 100083 China 2 Department of Cell Biology Capital University of Medical Sciences Beijing 100054 China To investigate whether the expression of exogenous heme oxygenase 1 HO 1 gene within vascular smooth muscle cells VSMC could protect the cells from free radical attack and inhibit cell proliferation we established an in vitro transfection of human HO 1 gene into rat VSMC mediated by a retroviral vector The results showed that the profound expression of HO 1 protein as well as HO activity was 1 8 and 2 0 fold increased respectively in the transfected cells compared to the non transfected ones The treatment of VSMC with different concentrations of H 2 O 2 led to the remarkable cell damage as indicated by survival rate and LDH leakage However the resistance of the HO 1 transfected VSMC against H 2 O 2 was significantly raised This protective effect was dramatically diminished when the transfected VSMC were pretreated with ZnPP IX a specific inhibitor of HO for 24 h In addition we found that the growth potential of the transfected cells was significantly inhibited directly by increased activity of HO 1 and this

    Original URL path: http://www.cell-research.com/arts.asp?id=1363 (2016-02-14)
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  • Cell Research
    Hong XIA Jian Fang GUI State Key laboratory of Freshwater Ecology and Biotechnology Wuhan Center for Developmental Biology Institute of Hydrobiology Chinese Academy of Sciences Wuhan 430072 China A systemic study was initiated to identify stage specific expression genes in fish embryogenesis by using suppression subtractive hybridization SSH technique In this study we presented a preliminary result on screen for stage specific expression genes between tail bud stage TBS and heartbeat beginning stage HBS in gynogenetic silver crucian carp Carassius auratus gibelio Two SSH plasmid libraries specific for TBS embryos and HBS embryos were constructed and stage specific expression genes were screened between the two stages 1963 TBS positive clones and 2466 HBS positive clones were sampled to PCR amplification and 1373 TBS and 1809 HBS PCR positive clones were selected to carry out dot blots 169 TBS dot blot positive clones and 272 HBS dot blot positive clones were sequenced Searching GenBank by using these nucleotide sequences indicated that most of the TBS dot blot positive clones could not be found homologous sequences in the database while known genes were mainly detected from HBS dot blot positive clones Of the 79 known genes 20 were enzymes or kinases involved

    Original URL path: http://www.cell-research.com/arts.asp?id=1364 (2016-02-14)
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  • Cell Research
    JI Feng LIU Er Qiu LI Yu Xian ZHU The National Laboratory of Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing 100871 China The nucleotide sequence deduced from the amino acid sequence of the scorpion insectotoxin AaIT was chemically synthesized and was expressed in Escherichia coli The authenticity of this in vitro expressed peptide was confirmed by N terminal peptide sequencing Two groups of bioassays artificial diet incorporation assay and contact insecticidal effect assay were carried out separately to verify the toxicity of this recombinant toxin At the end of a 24 h experimental period more than 60 of the testing diamondback moth Plutella xylostella larvae were killed in both groups with LC 50 value of 18 4 m M and 0 70 m M respectively Cytotoxicity assay using cultured Sf9 insect cells and MCF 7 human cells demonstrated that the toxin AaIT had specific toxicity against insect cells but not human cells Only 0 13 m M recombinant toxin was needed to kill 50 of cultured insect cells while as much as 1 3 m M toxin had absolutely no effect on human cells Insect cells produced obvious intrusions from their plasma membrane before

    Original URL path: http://www.cell-research.com/arts.asp?id=1365 (2016-02-14)
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  • Cell Research
    Department of Biophysics Health Science Center Peking University Beijing 100083 China AIM The existence and properties of alpha fetoprotein AFP receptor on the surface of NIH 3T3 cells and the effects of AFP on cellular signal transduction pathway were investigated METHODS The effect of AFP on the proliferation of NIH 3T3 cells was measured by incorporation of 3 H TdR Receptor binding assay of 125 I AFP was performed to detect the properties of AFP receptor in NIH 3T3 cells The influences of AFP on the cAMP i and the activities of protein kinase A PKA were determined Western blot was used to detect the change of K ras P21 protein expression RESULTS The proliferation of NIH 3T3 cells treated with 0 80 mg L of AFP was significantly enhanced The Scatchard analysis indicated that there were two classes of binding sites with K D of 2 722 10 9 M Bmax 12810 sites per cell and 8 931 10 8 M Bmax 119700 sites per cell respectively In the presence of AFP 20 mg L the content of cAMP and activities of PKA were significantly elevated The level of K ras P21 protein was upregulated by AFP at the

    Original URL path: http://www.cell-research.com/arts.asp?id=1366 (2016-02-14)
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  • Cell Research
    160 Get effective polyclonal antisera in one month Yuan Xin HU Ju Yuan GUO Lu SHEN Yan CHEN Zu Chuan ZHANG Yong Lian ZHANG 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 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

    Original URL path: http://www.cell-research.com/arts.asp?id=1367 (2016-02-14)
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  • Cell Research

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    Original URL path: /artsmore1.asp?id=118 (2016-02-14)


  • Cell Research
    metalloproteinases in Hydra ichael P SARRAS JR 1 Li YAN 2 Alexey LEONTOVICH 3 Jin Song ZHANG 4 1 Department of Anatomy and Cell Biology University of Kansas Medical Center Kansas City Kansas 66160 7400 USA 2 Centocor Malvern PA 19355 USA 3 Department of Experimental Pathology Mayo Clinic Rochester MN 55904 USA 4 Pharmaceutical Chemistry University of Kansas Lawrence KS 66047 USA Metalloproteinases have a critical role in a broad spectrum of cellular processes ranging from the breakdown of extracellular matrix to the processing of signal transduction related proteins These hydrolytic functions underlie a variety of mechanisms related to developmental processes as well as disease states Structural analysis of metalloproteinases from both invertebrate and vertebrate species indicates that these enzymes are highly conserved and arose early during metazoan evolution In this regard studies from various laboratories have reported that a number of classes of metalloproteinases are found in hydra a member of Cnidaria the second oldest of existing animal phyla These studies demonstrate that the hydra genome contains at least three classes of metalloproteinases to include members of the 1 astacin class 2 matrix metalloproteinase class and 3 neprilysin class Functional studies indicate that these metalloproteinases play diverse and

    Original URL path: http://www.cell-research.com/arts.asp?id=1349 (2016-02-14)
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  • Cell Research
    Shanghai Cancer Institute LN 2200 25 Xie tu Road Shanghai 200032 China The human RNA methyltransferase like 1 gene RNMTL1 is one of thirteen newly discovered genes within a 116 Kb segment of the chromosome 17p13 3 that suffers from a high frequent loss of heterozygosity in human hepatocellular carcinoma in China 1 5 To understand the molecular mechanisms underlying transcription control of the RNMTL1 gene in human cancers we decline using of the conventional approach where the cis elements bound by the known transcription factors are primary targets and carried out the systematic analyses to dissect the promoter structure and identify characterize the key cis elements that are responsible for its strong expression in cell The molecular approaches applied included 1 the primer extension for mapping of the transcription starts 2 the transient transfection reporter assays on a large number of deletion and site specific mutants of the promoter segment for defining the minimal promoter and the crucial elements within and 3 the electrophoresis mobility shift assay with specific antibodies for reconfirming the nature of the transcription factors and their cognate cis elements We have shown that the interaction of an ATF CREB element 38 to 31 and its

    Original URL path: http://www.cell-research.com/arts.asp?id=1350 (2016-02-14)
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