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  • Cell Research
    100 The expression and antigenicity identification of recombinant rat TGF β1 in bacteria GAO Chun Fang 1 Xian Tao KONG 1 Axel M GRESSNER 2 Ralf WEISKIRCHEN 2 1 Department of Laboratory Medicine Changzheng Hospital Second Military Medical University Shanghai 200003 China 2 Institute of Clinical Chemistry and Pathobiochemistry Central Laboratory RWTH University Hospital Pauwelsstr 30 D 52074 Aachen Germany In order to study structure function details of TGF β1 the recombinant mature form of rat TGF β 1 was expressed in bacteria Synthesis of the 112 amino acid carboxyl terminal part of TGF β1 amino acid 279 390 was controlled by an inducible gene expression system based on bacteriophage T7 RNA polymerase This system allowed an active and selective synthesis of recombinant TGF β1 The molecular weight of expressed TGF β1 monomer determined on SDS polyacrylamide gel under reducing conditions was about 13 kD Serial detergent washes combined with a single gel filtration purification step were sufficient to purify the expression product to homogeneity Amino terminal sequencing revealed that the N terminal of the recombinant protein was identical to the published data In Western blot analysis the recombinant polypeptide showed excellent antigenicity against polyclonal TGF b1 antibody The mature

    Original URL path: http://www.cell-research.com/arts.asp?id=1401 (2016-02-14)
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  • Cell Research
    C suppress the Trail induced apoptosis in Jurkat T cells GUO Ben Chang Yong Hua XU Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yue yang Road Shanghai 200031 China Trail a tumor necrosis factor related apoptosis inducing ligand is a novel potent endogenous activator of the cell death pathway through the activation of cell surface death receptors Trail R1 and Trail R2 Its role like FasL in activation induced cell death AICD has been demonstrated in immune system However the mechanism of Trail induced apoptosis remains unclear In this report the recombinant Trail protein was expressed and purified The apoptosis inducing activity and the regulation mechanism of recombinant Trail on Jurkat T cells were explored in vitro Trypan blue exclusion assay demonstrated that the recombinant Trail protein actively killed Jurkat T cells in a dose dependent manner Trail induced apoptosis in Jurkat T cells were remarkably reduced by Bcl 2 over expression in Bcl 2 gene transfected cells Treatment with PMA phorbol 12 myristate 13 acetate a PKC activator suppressed Trail induced apoptosis in Jurkat T cells The inhibition of apoptosis by PMA was abolished by pretreatment with Bis a PKC

    Original URL path: http://www.cell-research.com/arts.asp?id=1402 (2016-02-14)
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  • Cell Research
    Hong Zhi 1 Shu Fang WU 2 Zeng Hong TU 2 1 Department of Biological Regulation Weizmann Institute of Science Rehovot 76100 Israel Tel 00972 8 9343182 E mail hongzhi wicc weizmann ac il 7nbsp Fax 00972 8 9344116 2 State Key Laboratory of Drug Research Shanghai Institute of Materia Medica Chinese Academy of Sciences 294 Taiyuan Road Shanghai 200031 China A major problem which is poorly understood in the management of bladder cancer is low sensitivity to chemotherapy and high recurrence after transurethral resection Insulin like growth factor 1 receptor IGF 1R signaling plays a very important role in progression invasion and metastasis of bladder cancer cells In this study we investigated whether IGF 1R was involved in the growth stimulating activity and drug resistance of bladder cancer cells The results showed The mRNAs of IGF 1 IGF 2 and IGF 1R were strongly expressed in serum free cultured T24 cell line whereas normal urothelial cells did not express these factors receptors or only in trace levels T24 cell responded far better to growth stimulation by IGF 1 than did normal urothelial cells blockage of IGF1R by antisense oligodeoxynucleotide ODN significantly inhibited the growth of T24 cell and enhanced

    Original URL path: http://www.cell-research.com/arts.asp?id=1403 (2016-02-14)
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  • Cell Research
    Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China Gal a 1 3 Gal gal epitope is a carbohydrate epitope and synthesized in large amount by a 1 3 galactosyltransferase a 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 a 1 3 GT gene but produces a large amount of antibodies anti Gal which recognize Gal a 1 3 Gal structures specifically In this study a replication deficient recombinant adenoviral vector Ad5sGT containing pig a 1 3 GT cDNA was constructed and characterized Adenoviral vector mediated transfer of pig a 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

    Original URL path: http://www.cell-research.com/arts.asp?id=1404 (2016-02-14)
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  • Cell Research
    cells JIN Mei Lin 1 Ping ZHANG 3 Ming Xiao DING 2 Jing Ping YUN 3 Pei Fang CHEN 3 Yeuk Hon CHEN 3 Yin Qing CHEW 3 1 College of Animal Science and Veterinary Medicine Huazhong Agricultural University Wuhan 430070 China 2 College of Life Sciences Beijing University Beijing 100871 China 3 Chinese University of Hong Kong Hong Kong China The events of cell death and the expression of nuclear matrix protein NMP have been investigated in a promyelocytic leukemic cell line HL 60 induced with etoposide By means of TUNEL assay the nuclei displayed a characteristic morphology change and the amount of apoptotic cells increased early and reached maximun about 39 after treatment with etoposide for 2 h Nucleosomal DNA fragmentation was observed after treatment for 4 h The morphological change of HL 60 cells thus occurred earlier than the appearance of DNA ladder Total nuclear matrix proteins were analyzed by 2 dimensional gel electrophoresis Differential expression of 59 nuclear matrix proteins was found in 4 h etoposide treated cells Western blotting was then performed on three nuclear matrix acssociated proteins PML HSC70 and NuMA The expression of the suppressor PML protein and heat shock protein HSC70 were

    Original URL path: http://www.cell-research.com/arts.asp?id=1405 (2016-02-14)
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  • Cell Research
    China 2 Department of Public Health Second Military Medical University Shanghai 200433 China Effects of maternal dietary zinc deficiency on prenatal and postnatal brain development were investigated in ICR strain mice From d 1 of pregnancy E0 until postnatal d 20 P20 maternal mice were fed experimental diets that contained 1 mg Zn kg day severe zinc deficient SZD 5 mg Zn kg day marginal zinc deficient MZD 30 mg Zn kg day zinc adequately supplied ZA or 100 mg Zn kg day zinc supplemented ZS and pair fed PF Brains of offspring from these dietary groups were examined at various developmental stages for expression of nestin an intermediate filament protein found in neural stem cells and young neurons Immunocytochemistry showed nestin expression in neural tube 10 5 d post citrus dpc as well as in the cerebral cortex and neural tube from 10 5 dpc to postnatal d 10 P10 Nestin immunoreactivities in both brain and neural tube of those zinc supplemented control groups ZA ZS PF were stronger than those in zinc deficient groups SZD and MZD Western blot analysis confirmed that nestin levels in pooled brain extracts from each of the zinc supplemented groups ZA ZS PF

    Original URL path: http://www.cell-research.com/arts.asp?id=1406 (2016-02-14)
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  • Cell Research
    2001 142 148 HAL1 mediate salt adaptation in Arabidopsis thaliana YANG Su Xin 1 Yan Xiu ZHAO 2 Quan ZHANG 2 Yu Ke HE 1 Hui ZHANG 2 Da LUO 1 1 Institute of Plant Physiology Shanghai Institutes for Biological Sciences Chinese Academyof Science 300 Fenglin Road Shanghai 200032 China 2 Key Laboratory of Plant Stress Research Shandong Normal University Jinan 250014 China The yeast HAL1 gene was introduced into Arabidopsis thaliana by Agrobacterium tumefaciens mediated transformation with vacuum infiltration under the control of CaMV 35S promoter Thirty three individual kanamycin resistant plants were obtained from 75 000 seeds Southern blotting analysis indicated that HAL1 gene had been integrated into all of the transgenic plants genomes The copy number of HAL1 gene in transgenic plants was mostly 1 to 3 by Southern analysis Phenotypes of transgenic plants have no differences with wild type plants Several samples of transformants were self pollinated and progenies from transformed and non transformed plants controls were evaluated for salt tolerance and gene expression Measurement of concentrations of intracellular K and Na showed that transgenic lines were able to retain less Na than that of the control under salt stress Results from different tests indicated the

    Original URL path: http://www.cell-research.com/arts.asp?id=1407 (2016-02-14)
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  • Cell Research
    the spider insecticidal gene conferring resistance to leaffolder and striped stem borer HUANG Jian Qiu 1 Zhi Ming WEI 1 Hai Long AN 1 Yu Xian ZHU 2 1 National Laboratory of Plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institute for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China 2 National Laboratory of Plant Genetic Engineering and Protein Engineering College of Life Science Peking University Beijing 100871 China Immature embryos of rice varieties Xiushui11 and Chunjiang 11 precultured for 4d were infected and transformed by Agrobacterium tumefaciens strain EHA101 pExT 7 containing the spider insecticidal gene The resistant calli were transferred onto the differentiation medium and plants were regenerated The transformation frequency reached 56 72 measured as numbers of Geneticin G418 resistant calli produced and 36 60 measured as numbers of transgenic plants regenerated respectively PCR and Southern blot analysis of transgenic plants confirmed that the T DNA had been integrated into the rice genome Insect bioassays using T1 transgenic plants indicated that the mortality of the leaffolder Cnaphalocrasis medinalis after 7d of leaf feeding reached 38 61 and the corrected mortality of the striped stem borer Chilo suppressalis after 7d of leaf feeding reached 16

    Original URL path: http://www.cell-research.com/arts.asp?id=1408 (2016-02-14)
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