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  • Cell Research
    in B lymphocytes is a highly regulated event initiated by crosslinking of the B cell receptor BCR BCR engagement initiates several signaling events such as activation of PLCρ Ras and PI3K which generally speaking lead to survival However in the absence of survival signals CD40 or IL 4R engagement BCR crosslinking can also promote apoptotic signal transduction pathways such as activation of effector caspases expression of pro apoptotic genes and inhibition of pro survival genes The complex interplay between survival and death signals determines the B cell fate and consequently the immune response Genetic regulation of programmed cell death in Drosophila LEE Cheng Yu 1 2 Eric H BAEHRECKE 1 Full Text PDF Programmed cell death plays an important role in maintaining homeostasis during animal development and has been conserved in animals as different as nematodes and humans Recent studies of Drosophila have provided valuable information toward our understanding of genetic regulation of death Different signals trigger the novel death regulators rpr hid and grim that utilize the evolutionarily conserved iap and ark genes to modulate caspase function Subsequent removal of dying cells also appears to be accomplished by conserved mechanisms The similarity between Drosophila and human in cell death signaling pathways illustrate the promise of fruit flies as a model system to elucidate the mechanisms underlying regulation of programmed cell death DNA fragmentation in apoptosis ZHANG Jian hua Ming XU Full Text PDF Cleavage of chromosomal DNA into oligonucleosomal size fragments is an integral part of apoptosis Elegant biochemical work identified the DNA fragmentation factor DFF as a major apoptotic endonuclease for DNA fragmentation in vitro Genetic studies in mice support the importance of DFF in DNA fragmentation and possibly in apoptosis in vivo Recent work also suggests the existence of additional endonucleases for DNA degradation Understanding the roles of individual endonucleases in apoptosis and how they might coordinate to degrade DNA in different tissues during normal development and homeostasis as well as in various diseased states will be a major research focus in the near future ORIGINAL ARTICLES Effect of Bcl 2 and caspase 3 on calcium distribution in apoptosis of HL 60 cell ZHANG Min Hong Qing ZHANG Shao Bai XUE Full Text PDF Apoptosis manifests in two major execution programs downstream of the death signal the caspase pathway and organelle dysfunction An important antiapoptosis factor Bcl 2 protein contributes in caspase pathway of apoptosis Calcium an important intracellular signal element in cells is also observed to have changes during apoptosis which maybe affected by Bcl 2 protein We have previously reported that in Harringtonine HT induced apoptosis of HL 60 cells there s a change of intracellular calcium distribution moving from cytoplast especially Golgi s apparatus to nucleus and accumulating there with the highest concentration We report here that caspase 3 becomes activated in HT induced apoptosis of HL 60 cells which can be inhibited by overexpression of Bcl 2 protein No sign of apoptosis or intracellular calcium movement from Golgi s apparatus to nucleus in

    Original URL path: http://www.cell-research.com/artsmore.asp?id=126 (2016-02-14)
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  • Cell Research
    initiated by extracellular signals Protein tyrosine phosphatases which remove phosphate groups from tyrosine phosphorylated signaling molecules play equally important tyrosine roles as protein kinases in signal transduction SHP 2 a cytoplasmic SH2 domain containing protein tyrosine phosphatase is involved in the signaling pathways of a variety of growth factors and cytokines Recent studies have clearly demonstrated that this phosphatase plays an important role in transducing signal relay from the cell surface to the nucleus and is a critical intracellular regulator in mediating cell proliferation and differentiation ORIGINAL ARTICLES Temperature dependent expression of cdc2 and cyclin B1 in spermatogenic cells during spermatogenesis KONG Wei Hua 1 2 Zheng GU 1 Ji Ning LU 1 Jia Ke TSO 1 Full Text PDF p34 cdc2 and Cyclin B1 are key components of cell cycle controlling machine and are believed to play a fundamental role in gametogenesis It is also well known that in scrotal mammals spermatogenesis depends greatly on the maintenance of comparatively low temperature in the scrotum To investigate whether the expression of cdc2 and cyclin B1 in spermatogenic cells during spermatogenesis is actually a temperature dependent event in situ hybridization Western blotting and immunohistochemistry analysis were used to study the expression of cdc2 and cyclin B1 in normal and cryptorchid testis Results showed that the abdominal temperature had no significant influence on the transcription of cdc2 and cyclin B1 in the spermatogonia and pachytene diplotene primary spermatocytes but it blocked the translation of them Due to the deficiency of p34 cdc2 and Cyclin B1 the spermatogonia and pachytene diplotene primary spermatocytes were unable to form MPF hence they couldn t undergo karyokinesis The development of primary spermatocytes was arrested at the G2 to M phase transition We also found that testosterone could regulate the Cyclin B1 expression in spermatogenic cells Muscular injection of testosterone could recover spermatogenesis in the unilateral scrotal testis which was influenced by the contralateral cryptorchid testis but it could not salvage the spermatogenesis block in the cryptorchid testis Transgenic mice overexpressing γ aminobutyric acid transporter subtype I develop obesity MA Ying Hua 1 Jia Hua HU 1 Xiao Gang ZHOU 1 Ruo Wang ZENG 1 Zhen Tong MEI 2 Jian FEI 1 Li He GUO 1 Full Text PDF Transgenic mice ubiquitously overexpressing murine r 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 transgenic mice have slightly reduced spontaneous locomotive capacity and altered feeding pattern This 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 Genetic aberration in primary hepatocellular carcinoma correlation between p53 gene mutation and loss of hetero zygosity on chromosome 16q21 q23 and 9p21 p23 WANG Gang 1 2 Chang Hui HUANG 2 Yan ZHAO 2 Ling CAI 2 Ying WANG 2 Shi Jin XIU 2 Zheng Wen JIANG 2 Shuang YANG 2 Xin Tai ZHAO 1

    Original URL path: http://www.cell-research.com/artsmore.asp?id=125 (2016-02-14)
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  • Cell Research
    that benzylamine a membrane permeable weak base can mimick hydrogen peroxide H 2 O 2 to induce stomatal closure and butyric acid a membrane permeable weak acid can partly abolish the H 2 O 2 induced stomatal closure Confocal pH mapping with the probe 5 and 6 carboxy seminaphthorhodafluor 1 acetoxymethylester SNARF 1 AM revealed that H 2 O 2 leads to rapid changes in cytoplasmic and vacuolar pH in guard cells of Vicia faba L i e alkalinization of cytoplasmic areas occur red in parallel with a decrease of the vacuolar pH and that butyric acid pretreatment can abolish alkalinization of cytoplasmic areas and acidification of vacuolar areas of guard cells challenged with H 2 O 2 These results imply that the alkalinization of cytoplasm via efflux of cytosol protons into the vacuole in guard cells challenged with H 2 O 2 is important at an early stage in the signal cascade leading to stomatal closure Costimulation of resting B lymphocytes alters the IL 4 activated IRS2 signaling pathway in a STAT6 independent manner implications for cell survival and proliferation ZAMORANO Jose Ann E KELLY Jonathan AUSTRIAN Helen Y WANG Achsah D KEEGAN Full Text PDF IL 4 is an important B cell survival and growth factor IL 4 induced the tyrosine phosphorylation of IRS2 in resting B lymphocytes and in LPS or CD40L activated blasts Phosphorylated IRS2 coprecipitated with the p85 subunit of PI 3 kinase in both resting and activated cells By contrast association of phosphorylated IRS2 with GRB2 was not detected in resting B cells after IL 4 treatment although both proteins were expressed However IL 4 induced association of IRS2 with GRB2 in B cell blasts The pattern of IL 4 induced recruitment of p85 and GRB2 to IRS2 observed in B cells derived from STAT6 null mice was identical to that observed for normal mice While IL 4 alone does not induce activation of MEK a MEK1 inhibitor suppressed the IL 4 induced proliferative response of LPS activated B cell blasts These results demonstrate that costimulation of splenic B cells alters IL 4 induced signal transduction independent of STAT6 leading to proliferation Furthermore proliferation induced by IL 4 in LPS activated blasts is dependent upon the MAP kinase pathway p53 independent upregulation of p21 WAF1 in NIH 3T3 cells malignantly transformed by mot 2 TAKANO Syuichi 1 2 Renu WADHWA 3 Youji MITSUI 1 2 Sunil C KAUL 1 Full Text PDF Mot 2 protein is shown to interact with p53 and inhibit its transcriptional activation function Mot 2 overexpressing stable clones of NIH 3T3 cells were malignantly transformed however they had a high level of expression of a p53 downstream gene p21 WAF1 The present study was undertaken to elucidate possible molecular mechanism s of such upregulation An increased level of p21 WAF1 expression was detected in stable transfectants although an exogenous reporter gene driven by p21 WAF1 promoter exhibited lower activity in these cells suggesting that some post transcriptional mechanism contributes to

    Original URL path: http://www.cell-research.com/artsmore.asp?id=124 (2016-02-14)
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  • Cell Research
    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 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 Altered expression of nuclear matrix proteins in etoposide induced apoptosis in HL 60 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 Full Text PDF 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 significantly upregulated after etoposide treatment while NuMA a nuclear mitotic apparatus protein was down regulated These results demonstrate that significant biochemical alterations in nuclear matrix proteins take place during the apoptotic process Maternal zinc deficiency impairs brain nestin expression in prenatal and postnatal mice WANG Fu Di 1 2 Wei BIAN 1 Ling Wei KONG 1 Fa Ji ZHAO 2 Jun Sheng GUO 2 Nai He JING 1 Full Text PDF 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

    Original URL path: http://www.cell-research.com/artsmore.asp?id=123 (2016-02-14)
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  • Cell Research
    acids in length were used to screen with the serum from a hepatitis B virus infected patient in the recovery phase After 3 rounds of biopanning the positive phages were confirmed by competitive ELISA using HBsAg P33 Two phagotopes were identified and one of them was confirmed as mimotope by competition experiment Based on the mimotpe a multiple antigenic peptide with four branches was synthesized by solid phase peptide synthesis The antiginicity and specificity of the synthesized antigen was tested in BALB c mice compared with the native epitope based antigen The results showed that the mimotope based antigen could evoke higher titer of antibodies with the same specificity of the epitope based antigen Those findings indicate mimotopes can be used in antigen and vaccine design Cloning and characterization of a novel gene C17orf25 from the deletion region on chromosome 17p13 3 in hepatocelular carcinoma QIN Wen Xin 1 Da Fang WAN 1 Fen Yong SUN 1 Ping Ping ZHANG 1 Li Wei HAN 1 Yi HUANG 1 Hui Qiu JIANG 1 Xin Tai ZHAO 1 Mei HE 2 Yun YE 2 Wen Ming CONG 3 Meng Chao WU 3 Li Sheng ZHANG 4 Nan Wu YANG 4 Jian Ren GU 1 Full Text PDF Using a combination of hybridization of PAC to a cDNA library and RACE technique we isolated a novel cDNA designated as C17orf25 Chromosome 17 open reading frame 25 previously named it HC71A from the deletion region on chromosome 17p13 3 The cDNA encodes a protein of 313 amino acids with a calculated molecular mass of 34 8 kDa C17orf25 is divided into 10 exons and 9 introns spanning 23 kb of genomic DNA Northern blot analysis showed that the mRNA expression of C17orf25 was decreased in hepatocellular carcinoma samples as compared to adjacent noncancerous liver tissues from the same patients The transfection of C17orf25 into the hepatocellular carcinoma cell SMMC7721 and overexpression could inhibit the cell growth The above results indicate that C17orf25 is a novel human gene and the cloning and preliminary characterization of C17orf25 is a prerequisite for further functional analysis of this novel gene in human hepatocellular carcinoma TNF receptor associated factor 2 binding site is involved in TNFR75 dependent enhancement of TNFR55 induced cell death LU Fang Jia FANG Chang Qing CHEN Full Text PDF TNF recepter 55 is the main mediator of TNF induced apoptosis TNF receptor 75 dependent induction or enhancement of cytotoxicity has been explained by intracellular signaling ligand passing or induction of endogenous TNF To study the function of human TNF receptor 75 hTR75 and the interaction between human TNF receptor 55 hTR55 and hTR75 in hTNF a induced cytotoxicity HEp 2 cells were transfected with bicistronic expression vector of h TR 75 and its deletion mutants genes hTNF a induced cytotoxicity was determined by crystal violet colorimetric method The expression of hTR75 and its deletion mutants in HEp 2 cells was demonstrated by RT PCR and indirect ELISA We found that the overexpressed hTR75 could significantly

    Original URL path: http://www.cell-research.com/artsmore.asp?id=122 (2016-02-14)
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  • Cell Research
    confirmed that T DNA had been intergrated into the A pseudoalhagi genome Acetosyringone AS was found to be vital for successful transformation of A pseudoalhagi Overexpression of a novel gene Cms1 can rescue the growth arrest of a Saccharomyces cerevisiae mcm10 suppressor WANG Ji Wu Jia Rui WU Full Text PDF MCM10 protein is an essential replication factor involved in the initiation of DNA replication A mcm10 mutant mcm10 1 of budding yeast shows a growth arrest at 37oC In the present work we have isolated a mcm10 1 suppressor strain which grows at 37oC Interestingly this mcm10 1 suppressor undergoes cell cycle arrest at 14oC A novel gene YLR003c is identified by high copy complementation of this suppressor We called it as Cms1 Complementation of Mcm10 Suppressor Furthermore the experiments of transformation show that cells of mcm10 1 suppressor with high copy plasmid but not low copy plasmid grow at 14oC indicating that overexpression of Cms1 can rescue the growth arrest of this mcm10 suppressor at non permissive temperature These results suggest that CMS1 protein may functionally interact with MCM10 protein and play a role in the regulation of DNA replication and cell cycle control High IFN α expression is associated with the induction of experimental autoimmune uveitis EAU in Fischer 344 rat Yong Jun Lei ZANG Ya Di WU Bing SUN Full Text PDF Th1 response plays a crucial role in determining pathogenesis of organ specific autoimmune diseases It is believed that both IL 12 and INF a are initiators to regulate Th1 response In our experimental autoimmune uveitis EAU model both Lewis and Fischer 344 rats share the same MHC class II molecules while Lewis rat is EAU susceptible and Fischer 344 rat is EAU resistant However under the same condition of immunization if pertussis toxin PTX was injected intraperitoneally as an additional adjuvant Fischer 344 rat can develop EAU In this study we investigate which mechanisms are involved in the induction of EAU in CFA R16 PTX treated CRP treated Fischer 344 rats In vivo and in vitro data demonstrated that Th1 cytokine IFN g mRNA expression was significantly increased in disease target tissue eyes and in draining lymph node cells of CRP treated Fischer 344 rat When IL 12 and IFN a mRNA expression were compared in the experimental groups only IFN a mRNA expression was associated with EAU development To distinguish the sources of IFN a producing cells it was observed that IFN a expression was mainly produced by macrophages It was further confirmed that normal macrophage from Fischer 344 rat was able to produce significant IFN a in the presence of PTX The data strongly suggested that IFN a might be involved in initiating Th1 cell differentiation and in turn contribute to the induction of EAU High IFN a expression induced by PTX may represent a novel pathway to initiate Th1 response in Fischer 344 rat Function of GATA transcription factors in hydroxyurea induced HEL cells ZHANG Shu Bing Qi Ye HE Hui

    Original URL path: http://www.cell-research.com/artsmore.asp?id=121 (2016-02-14)
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  • Cell Research
    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 Four of 69 cDNA fragments 3 up regulated named L1 L2 and L3 and 1 down regulated named M1 in leiomyoma were confirmed by Northern analysis 2 Sequence comparison and Northern analysis proved that L1 is exactly the human ribosomal protein S19 3 It was present ubiquitously in 13 tissues tested but in various levels and even in different size 4 L1 was highly expressed in parotidean cystadenocarcinoma pancreatic cancer and breast cancer examined No mutations have been found in human uterine leiomyomas n 6 CONCLUSIONS hRPS19 overexpression migh be a universal signal in rapid cell growth tissues The inhibition of lung cancer cell growth by intracellular immunization with LC 1 ScFv Liang CHEN Gang LI Lei TANG Jue WANG Xi Rui GE Full Text PDF A monoclonal antibody LC 1 recognizing lung cancer associated common antigens was obtained in authors laboratory Its single chain Fv fragment ScFv named LC 1 ScFv was constructed based on recombinant phage displayed techniques For expression on cell membrane LC 1 ScFv was cloned into pDisplay vector which directed the cloned gene to express as cell membrane bound protein The resulting plasmid was sequenced and then introduced by the lipofectin method into a lung adenocarcinoma cell line SPC A 1 G418 resistant cells were obtained by G418 selection After transfection LC 1 ScFv expression was observed by Western blot analysis and the expression of cognate antigens was down regulated as shown in ELISA assay SPC A 1 pDisplay ScFv cells grew in vitro at lower speed than the control intact cells and the cells transfected with vacant vector Flow cytometry analysis detected a substantial increase in G 1 phase and decrease in S phase in population of SPC A 1 pDisplay ScFv cells compared to SPC A 1 and SPC A 1 pDisplay cells Semi quantitative RT PCR analysis showed that c myc expression was down regulated in SPC A 1 pDisplay ScFv cells It seems that the antigens recognized by LC 1 may be in some way involved in a growth stimulating pathway and the antibody blocking of the function of the antigens shut down the pathway and thus down regulate the expression of c myc and growth of the cells Effects of cisplatin on telomerase activity and telomere length in BEL 7404 human hepatoma cells Ru Gang ZHANG Ru Ping ZHANG Xing Wang WANG Hong XIE Full Text PDF Telomerase activity was inhibited in a dose and time dependent manner with the treatment of cisplatin for 24 48 or 72 h in a concentration ranged from 0 8 to 50 μM in BEL 7404 human hepatoma cells There were no changes in expression pattern of three telomerase subunits its catalytic reverse transcriptase subunit hTERT its RNA component hTR or the associated

    Original URL path: http://www.cell-research.com/artsmore.asp?id=120 (2016-02-14)
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  • Cell Research
    of heme oxygenase 1 protects smooth muscle cells against oxidative injury and inhibits cell proliferation Min ZHANG 1 Bao Hui ZHANG 1 Li CHEN 2 Wei AN 2 Full Text PDF 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 effect might be related to decreased phosphorylation of MAPK These results suggest that the overexpression of introduced hHO 1 is potentially able to reduce the risk factors of atherosclerosis partially due to its cellular protection against oxidative injury and to its inhibitory effect on cellular proliferation Screen for stage specific expression genes between tail bud stage and heartbeat beginning stage in embryogenesis of gynogenetic silver crucian carp Yao Hua SHI Jun LIU Jian Hong XIA Jian Fang GUI Full Text PDF 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 in important metabolism of embryonic development Moreover specific expressions of partial genes were further confirmed by virtual northern blots This study is the first step for making a large attempt to study temporal and spatial control of gene expression in the

    Original URL path: http://www.cell-research.com/artsmore.asp?id=119 (2016-02-14)
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