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  • Cell Research
    interactions could reveal new potential targets for pharmacological interventions in respiratory epithelium carcinogenesis ORIGINAL ARTICLES In vitro derivation of functional insulin producing cells from human embryonic stem cells Wei Jiang 1 Yan Shi 1 5 Dongxin Zhao 1 Song Chen 1 Jun Yong 1 Jing Zhang 2 Tingting Qing 1 Xiaoning Sun 1 3 Peng Zhang 1 Mingxiao Ding 1 Dongsheng Li 4 and Hongkui Deng 1 2 3 Cell Research 2007 17 333 344 doi 10 1038 cr 2007 28 published online 10 April 2007 Full Text PDF The capacity for self renewal and differentiation of human embryonic stem ES cells makes them a potential source for generation of pancreatic beta cells for treating type I diabetes mellitus Here we report a newly developed and effective method carried out in a serum free system which induced human ES cells to differentiate into insulin producing cells Activin A was used in the initial stage to induce definitive endoderm differentiation from human ES cells as detected by the expression of the definitive endoderm markers Sox17 and Brachyury Further all trans retinoic acid RA was used to promote pancreatic differentiation as indicated by the expression of the early pancreatic transcription factors pdx1 and hlxb9 After maturation in DMEM F12 serum free medium with bFGF and nicotinamide the differentiated cells expressed islet specific markers such as C peptide insulin glucagon and glut2 The percentage of C peptide positive cells exceeded 15 The secretion of insulin and C peptide by these cells corresponded to the variations in glucose levels When transplanted into renal capsules of Streptozotocin STZ treated nude mice these differentiated human ES cells survived and maintained the expression of beta cell marker genes including C peptide pdx1 glucokinase nkx6 1 IAPP pax6 and Tcf1 Thirty percent of the transplanted nude mice exhibited apparent restoration of stable euglycemia and the corrected phenotype was sustained for more than six weeks Our new method provides a promising in vitro differentiation model for studying the mechanisms of human pancreas development and illustrates the potential of using human ES cells for the treatment of type I diabetes mellitus Xom interacts with and stimulates transcriptional activity of LEF1 TCFs implications for ventral cell fate determination during vertebrate embryogenesis Hong Gao 1 2 Bin Wu 1 Roger Giese 2 and Zhenglun Zhu 1 Cell Research 2007 17 345 356 doi 10 1038 cr 2007 20 published online 3 April 2007 Full Text PDF LEF1 TCFs are high mobility group box containing transcriptional factors mediating canonical Wnt β catenin signaling during early embryogenesis and tumorigenesis β Catenin forms a complex with LEF1 TCFs and transactivates LEF1 TCF mediated transcriptions during dorsalization Although LEF mediated transcription is also implicated in ventralization the underlying molecular mechanism is not well understood Using the vertebrate Xenopus laevis model system we found that Xom which is a ventralizing homeobox protein with dual roles of transcriptional activation and repression forms a complex with LEF1 TCF through its homeodomain and transactivates LEF1 TCF mediated transcription through its

    Original URL path: http://www.cell-research.com/artsmore.asp?id=75 (2016-02-14)
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  • Cell Research
    genes that were differentially regulated during fiber development Principal component analysis PCA using expressed genes as variables divided fiber samples into four groups which are diagnostic of developmental stages Similar grouping results are also found if we use non polar or polar metabolites as variables for PCA of developing fibers Auxin signaling wall loosening and lipid metabolism are highly active during fiber elongation whereas cellulose biosynthesis is predominant and many other metabolic pathways are downregulated at the secondary cell wall synthesis stage Transcript and metabolite profiles and enzyme activities are consistent in demonstrating a specialization process of cotton fiber development toward cellulose synthesis These data demonstrate that cotton fiber cell at a certain stage has its own unique feature and developmental stages of cotton fiber cells can be distinguished by their transcript and metabolite profiles During the secondary cell wall synthesis stage metabolic pathways are streamed into cellulose synthesis A novel heterodimeric cytokine consisting of IL 17 and IL 17F regulates inflammatory responses Seon Hee Chang 1 and Chen Dong 1 Cell Research 2007 17 435 440 doi 10 1038 cr 2007 35 published online 24 April 2007 Full Text PDF CD4 helper T TH cells play crucial roles in immune responses Recently a novel subset of TH cells termed TH IL 17 TH17 or inflammatory TH THi has been identified as critical mediators of tissue inflammation These cells produce IL 17 also called IL 17A and IL 17F two most homologous cytokines sharing similar regulations Here we report that when overexpressed in 293T cells IL 17 and IL 17F form not only homodimers but also heterodimers which we name as IL 17A F Fully differentiated mouse THi cells also naturally secrete IL 17A F as well as IL 17 and IL 17F homodimeric cytokines Recombinant IL 17A F protein exhibits intermediate levels of potency in inducing IL 6 and KC CXCL1 as compared to homodimeric cytokines IL 17A F regulation of IL 6 and KC expression is dependent on IL 17RA and TRAF6 Thus IL 17A F cytokine represents another mechanism whereby T cells regulate inflammatory responses and may serve as a novel target for treating various immune mediated diseases Bcl 2 cleavages at two adjacent sites by different caspases promote cisplatin induced apoptosis Jianbei Zhu 1 Ying Yang 1 and Jiarui Wu 1 2 Cell Research 2007 17 441 448 doi 10 1038 cr 2007 36 published online 24 April 2007 Full Text PDF The protein encoded by bcl 2 proto oncogene plays an important role in the mitochondria mediated apoptotic pathway Although the general role of Bcl 2 is anti apoptotic previous work showed that Bcl 2 fragments cleaved by caspases could promote apoptotic process We report herein that Bcl 2 protein was cleaved to produce two fragments of around 23 kDa in human hepatocarcinoma BEL 7404 cells or in Bcl 2 overexpressing CHO cells induced by cisplatin Treating cells with the general caspase inhibitor z VAD fmk blocked the induced cleavage of Bcl 2 Mutagenesis analyses

    Original URL path: http://www.cell-research.com/artsmore.asp?id=74 (2016-02-14)
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  • Cell Research
    Bcl 2 is required for the progression of LNCaP prostate cancer cells from an androgen dependent to an androgen independent growth stage The mRNA and protein levels of Bcl 2 are significantly increased in androgen independent prostate cancer cells shRNA mediated gene silencing of Bcl 2 in androgen independent prostate cancer cells promotes UV induced apoptosis and suppresses the growth of prostate tumors in vivo Growing androgen dependent cells under androgen deprivation conditions results in formation of androgen independent colonies and the transition from androgen dependent to androgen independent growth is blocked by ectopic expression of the Bcl 2 antagonist Bax or Bcl 2 shRNA Thus our results demonstrate that Bcl 2 is not only critical for the survival of androgen independent prostate cancer cells but is also required for the progression of prostate cancer cells from an androgen dependent to an androgen independent growth stage Cilia containing 9 2 structures grown from immortalized cells Ming Zhang 1 2 and Jose G Assouline 3 Cell Research 2007 17 537 545 doi 10 1038 sj cr 7310151 published online 17 April 2007 Full Text PDF Cilia depend on their highly differentiated structure a 9 2 arrangement to remove particles from the lung and to transport reproductive cells Immortalized cells could potentially be of great use in cilia research Immortalization of cells with cilia structure containing the 9 2 arrangement might be able to generate cell lines with such cilia structure However whether immortalized cells can retain such a highly differentiated structure remains unclear Here we demonstrate that 1 using E1a gene transfection tracheal cells are immortalized 2 interestingly in a gel culture the immortalized cells form spherical aggregations within which a lumen is developed and 3 surprisingly inside the aggregation cilia containing a 9 2 arrangement grow from the cell s apical pole and protrude into the lumen These results may influence future research in many areas such as understanding the mechanisms of cilia differentiation cilia generation in other existing cell lines cilia disorders generation of other highly differentiated structures besides cilia using the gel culture immortalization of other ciliated cells with the E1a gene development of cilia motile function and establishment of a research model to provide uniform ciliated cells Early Growth Response gene 1 Egr 1 regulates HSV 1 ICP4 and ICP22 gene expression Gautam R Bedadala Rajeswara C Pinnoji and Shao Chung V Hsia Cell Research 2007 17 546 555 doi 10 1038 cr 2007 44 published online 15 May 2007 Full Text PDF The molecular mechanisms mediating herpes simplex virus type 1 HSV 1 gene silencing during latent infection are not clear Five copies of early growth response gene 1 Egr 1 binding elements were identified in the intron of HSV 1 ICP22 infected cell protein No 22 gene leading to the hypothesis that Egr 1 binds to the viral genome and regulates the viral gene expression Transient co transfection assays indicated that Egr 1 negatively regulated the transcription of both full length and intron removed ICP22

    Original URL path: http://www.cell-research.com/artsmore.asp?id=73 (2016-02-14)
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  • Cell Research
    novel integrator of spindle checkpoint signaling However it is unclear how NEK2A regulates kinetochore microtubule attachment in mitosis Here we show that NEK2A phosphorylates human Sgo1 and such phosphorylation is essential for faithful chromosome congression in mitosis NEK2A binds directly to HsSgo1 in vitro and co distributes with HsSgo1 to the kinetochore of mitotic cells Our in vitro phosphorylation experiment demonstrated that HsSgo1 is a substrate of NEK2A and the phosphorylation sites were mapped to Ser 14 and Ser 507 as judged by the incorporation of 32 P Although such phosphorylation is not required for assembly of HsSgo1 to the kinetochore expression of non phosphorylatable mutant HsSgo1 perturbed chromosome congression and resulted in a dramatic increase in microtubule attachment errors including syntelic and monotelic attachments These findings reveal a key role for the NEK2A mediated phosphorylation of HsSgo1 in orchestrating dynamic kinetochore microtubule interaction We propose that NEK2A mediated phosphorylation of human Sgo1 provides a link between centromeric cohesion and spindle microtubule attachment at the kinetochores Interactions of primary neuroepithelial progenitor and brain endothelial cells distinct effect on neural progenitor maintenance and differentiation by soluble factors and direct contact Miguel A Gama Sosa 1 3 Rita De Gasperi 1 3 Anne B Rocher 2 Gissel M Perez 1 3 Keila Simons 1 3 Daniel E Cruz 1 3 Patrick R Hof 2 and Gregory A Elder 1 3 Cell Research 2007 17 619 626 doi 10 1038 cr 2007 53 published online 26 June 2007 Full Text PDF Neurovascular interactions are crucial for the normal development of the central nervous system To study such interactions in primary cultures we developed a procedure to simultaneously isolate neural progenitor and endothelial cell fractions from embryonic mouse brains Depending on the culture conditions endothelial cells were found to favor maintenance of the neuroprogenitor phenotype through the production of soluble factors or to promote neuronal differentiation of neural progenitors through direct contact These apparently opposing effects could reflect differential cellular interactions needed for the proper development of the brain Downregulation of CD4 CD25 regulatory T cells may underlie enhanced Th1 immunity caused by immunization with activated autologous T cells Qi Cao 1 2 Li Wang 1 2 Fang Du 1 2 Huiming Sheng 1 2 Yan Zhang 1 2 Juanjuan Wu 1 2 Baihua Shen 1 2 Tianwei Shen 1 2 Jingwu Zhang 1 2 Dangsheng Li 3 and Ningli Li 1 2 Cell Research 2007 17 627 637 doi 10 1038 cr 2007 46 published online 12 June 2007 Full Text PDF Regulatory T cells Treg play important roles in immune system homeostasis and may also be involved in tumor immunotolerance by suppressing Th1 immune response which is involved in anti tumor immunity We have previously reported that immunization with attenuated activated autologous T cells leads to enhanced anti tumor immunity and upregulated Th1 responses in vivo However the underlying molecular mechanisms are not well understood Here we show that Treg function was significantly downregulated in mice that received immunization of attenuated activated

    Original URL path: http://www.cell-research.com/artsmore.asp?id=72 (2016-02-14)
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  • Cell Research
    Fe S protein 1 of NADH dehydrogenase dihydrolipoamide acetyltransferase or E2 component of the pyruvate dehydrogenase complex and lactate dehydrogenase 2 Further real time quantitative PCR analyses confirmed that the levels of the corresponding mRNAs were also remarkably reduced Consistent with these findings lower ATP levels and an impaired ability to convert glucose into ATP were also observed in the hippocampus of chronically treated mice Opioid antagonist naltrexone administrated concomitantly with morphine significantly suppressed morphine withdrawal jumping and reversed the downregulation of these proteins Acute exposure to morphine also produced robust morphine withdrawal jumping and significant memory impairment but failed to decrease the expression of these three proteins Intrahippocampal injection of D glucose before morphine administration significantly enhanced ATP levels and suppressed morphine withdrawal jumping and memory impairment in acute morphine treated but not in chronic morphine treated mice Intraperitoneal injection of high dose of D glucose shows a similar effect on morphine induced withdrawal jumping as the central treatment Taken together our results suggest that reduced expression of the three metabolic enzymes in the hippocampus as a result of chronic morphine treatment contributes to the development of drug induced symptoms such as morphine withdrawal jumping and memory impairment Kinetochore dynein generates a poleward pulling force to facilitate congression and full chromosome alignment Yan Li 1 Wei Yu 1 Yun Liang 1 and Xueliang Zhu 1 Cell Research 2007 17 701 712 doi 10 1038 cr 2007 65 published online 7 August 2007 Full Text PDF For proper chromosome segregation all kinetochores must achieve bipolar microtubule MT attachment and subsequently align at the spindle equator before anaphase onset The MT minus end directed motor dynein dynactin binds kinetochores in prometaphase and has long been implicated in chromosome congression Unfortunately inactivation of dynein usually disturbs spindle organization thus hampering evaluation of its kinetochore roles Here we specifically eliminated kinetochore dynein dynactin by RNAi mediated depletion of ZW10 a protein essential for kinetochore localization of the motor Time lapse microscopy indicated markedly reduced congression efficiency though congressing chromosomes displayed similar velocities as in control cells Moreover cells frequently failed to achieve full chromosome alignment despite their normal spindles Confocal microcopy revealed that the misaligned kinetochores were monooriented or unattached and mostly lying outside the spindle suggesting a difficulty to capture MTs from the opposite pole Kinetochores on monoastral spindles were dispersed farther away from the pole and exhibited only mild oscillation Furthermore inactivating dynein by other means generated similar phenotypes Therefore kinetochore dynein produces on monooriented kinetochores a poleward pulling force which may contribute to efficient bipolar attachment by facilitating their proper microtubule captures to promote congression as well as full chromosome alignment OsRRM a Spen like rice gene expressed specifically in the endosperm Shi Yan Chen 1 2 Zong Yang Wang 1 and Xiu Ling Cai 1 Cell Research 2007 17 713 721 doi 10 1038 cr 2007 43 published online 8 May 2007 Full Text PDF We used the promoter trap technique to identify a rice plant named 107 in

    Original URL path: http://www.cell-research.com/artsmore.asp?id=71 (2016-02-14)
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  • Cell Research
    RNAi adipocytes Glut 4 was localized predominantly in non caveolar lipid rafts After the removal of insulin caveolae localized Glut 4 was internalized faster than non caveolar lipid raft associated Glut 4 The internalization of Glut 4 from plasma membrane was significantly decreased in Cav 1 RNAi adipocytes These results suggest that insulin stimulated Glut 4 translocation and glucose uptake are caveolae independent events Caveolae play a role in the internalization of Glut 4 from plasma membrane after the removal of insulin Quantitative and qualitative in vitro analysis of the stem cell potential of hematopoietic cells purified from murine skeletal muscle Celine Haond 1 Françoise Farace 1 Martine Guillier 1 Yann Lécluse 1 Frederic Mazurier 2 William Vainchenker 1 and Ali G Turhan 1 Cell Research 2007 17 783 791 doi 10 1038 cr 2007 74 published online 11 September 2007 Full Text PDF The murine skeletal muscle contains hematopoietic stem cells but this potential has so far not been studied quantitatively or qualitatively in vitro To quantify the hematopoietic stem cell potential we have used highly purified SP CD45 cells in long term culture initiating cell LTC IC assays The SP CD45 cell population purified from murine muscle was found to have significant stem cell activity with an LTC IC frequency of 1 640 Single cell sorted SP CD45 cells from muscle exhibited robust proliferative activity in vitro at day 16 380 fold amplification especially after culture with OP 9 layers that also support embryonic stem cells Amplified cell populations originating from single cells exhibited multilineage differentiation ability with evidence of myeloid lymphoid and NK cell markers Thus our results demonstrate that hematopoietic stem cells that can be quantified by LTC IC assays exist in the murine skeletal muscle and show also for the first time at the single cell level that these cells exhibit multilineage differentiation ability and major proliferative potential Activation of paternally expressed imprinted genes in newly derived germline competent mouse parthenogenetic embryonic stem cell lines Hua Jiang 1 2 3 Bowen Sun 1 2 Weicheng Wang 1 2 Zhihong Zhang 1 Furong Gao 1 2 3 Guilai Shi 1 2 3 Bing Cui1 Xiangyin Kong 1 Zhao He 4 Xiaoyan Ding 4 Ying Kuang 5 Jian Fei 5 Yi Juan Sun 6 Yun Feng 6 and Ying Jin 1 2 Cell Research 2007 17 792 803 doi 10 1038 cr 2007 70 published online 4 September 2007 Full Text PDF Parthenogenetic embryonic stem pES cells provide a valuable in vitro model system for studying the molecular mechanisms that underlie genomic imprinting However the pluripotency of pES cells and the expression profiles of paternally expressed imprinted genes have not been fully explored In this study three mouse pES cell lines were established and the differentiation potential of these cells in extended culture was evaluated The undifferentiated cells had a normal karyotype and homozygous genome and expressed ES cell specific molecular markers The cells remained undifferentiated after more than 50 passages and exhibited pluripotent differentiation capacity All

    Original URL path: http://www.cell-research.com/artsmore.asp?id=70 (2016-02-14)
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  • Cell Research
    histone demethylases has proved that histone methylation is a reversible process Through a candidate approach we have biochemically identified JMJD3 as an H3K27 demethylase Transfection of JMJD3 into HeLa cells caused a specific reduction of trimethyl H3K27 but had no effect on di and monomethyl H3K27 or histone lysine methylations on H3K4 and H3K9 The enzymatic activity requires the JmjC domain and the conserved histidine that has been suggested to be important for a cofactor binding In vitro biochemical experiments demonstrated that JMJD3 directly catalyzes the demethylation In addition we found that JMJD3 is upregulated in prostate cancer and its expression is higher in metastatic prostate cancer Thus we identified JMJD3 as a demethylase capable of removing the trimethyl group from histone H3 lysine 27 and upregulated in prostate cancer Rig I mice develop colitis associated with downregulation of Gαi2 Yi Wang 1 Hong Xin Zhang 1 Yue Ping Sun 1 Zi Xing Liu 1 Xue Song Liu 1 Long Wang 2 3 Shun Yuan Lu 2 3 Hui Kong 3 Qiao Ling Liu 3 Xi Hua Li 1 Zhen Yu Lu 1 Sai Juan Chen 2 Zhu Chen 2 Shi San Bao 4 Wei Dai 5 and Zhu Gang Wang 1 2 3 4 Cell Research 2007 17 858 868 doi 10 1038 cr 2007 81 published online 25 September 2007 Full Text PDF RIG I retinoid acid inducible gene I a putative RNA helicase with a cytoplasmic caspase recruitment domain CARD was identified as a pattern recognition receptor PRR that mediates antiviral immunity by inducing type I interferon production To further study the biological function of RIG I we generated Rig I mice through homologous recombination taking a different strategy to the previously reported strategy Our Rig I mice are viable and fertile Histological analysis shows that Rig I mice develop a colitis like phenotype and increased susceptibility to dextran sulfate sodium induced colitis Accordingly the size and number of Peyer s patches dramatically decreased in mutant mice The peripheral T cell subsets in mutant mice are characterized by an increase in effector T cells and a decrease in naïve T cells indicating an important role for Rig I in the regulation of T cell activation It was further found that Rig I deficiency leads to the downregulation of G protein αi2 subunit Gαi2 in various tissues including T and B lymphocytes By contrast upregulation of Rig I in NB4 cells that are treated with ATRA is accompanied by elevated G α i2 expression Moreover G α i2 promoter activity is increased in co transfected NIH3T3 cells in a Rig I dose dependent manner All these findings suggest that Rig I has crucial roles in the regulation of G α i2 expression and T cell activation The development of colitis may be at least in part associated with downregulation of G α i2 and disturbed T cell homeostasis Induction of the LRP16 gene by estrogen promotes the invasive growth of Ishikawa human endometrial cancer cells through the downregulation

    Original URL path: http://www.cell-research.com/artsmore.asp?id=69 (2016-02-14)
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  • Cell Research
    with PI 3 P PI 4 P and PI 3 5 P2 as substrates in vitro and a mutation in the catalytic core of the Sac domain abolishes its enzymatic activity The expression of rSac3 is upregulated during nerve growth factor NGF stimulated PC12 cell neuronal differentiation and overexpression of this protein promotes neurite outgrowth in PC12 cells Conversely inhibition of rSac3 expression by antisense oligonucleotides reduces neurite outgrowth of NGF stimulated PC12 cells and the active site mutation of rSac3 eliminates its neurite outgrowth promoting activity These results indicate that rSac3 promotes neurite outgrowth in differentiating neurons through its PIPPase activity suggesting that Sac domain PIPPase proteins may participate in forward membrane trafficking from the endoplasmic reticulum and Golgi complex to the plasma membrane and may function as regulators of this crucial process of neuronal cell growth and differentiation Andrographolide inhibits NF κB activation and attenuates neointimal hyperplasia in arterial restenosis Yu Jiu Wang 1 Jin Tao Wang 2 Quan Xin Fan 1 and Jian Guo Geng 2 Cell Research 2007 17 933 941 doi 10 1038 cr 2007 89 published online 16 October 2007 Full Text PDF The NF κB transcription factors modulate the expression of tissue factor TF E selectin CD62E and vascular cell adhesion molecule 1 VCAM 1 which are essential for thrombosis and inflammation We have previously shown that andrographolide Andro covalently modifies the reduced cysteine 62 of p50 a major subunit of NF κB transcription factors thus blocking the binding of NF κB transcription factors to the promoters of their target genes preventing NF κB activation and inhibiting inflammation in vitro and in vivo Here we report that Andro but not its inactive structural analog 4H Andro significantly suppressed the proliferation of arterial neointima κ60 reduction in a murine model of arterial restenosis Consistently p50 mice manifested attenuated neointimal hyperplasia upon arterial ligation Notably the same dosage of Andro did not further reduce neointimal formation in p50 mice which implicates the specificity of Andro on p50 for treating experimental arterial restenosis The upregulation of NF κB target genes including TF E selectin and VCAM 1 and the increased deposition of leukocytes mainly CD68 macrophages were clearly detected within the injured arterial walls all of which were significantly abolished by treatment with Andro or genetic deletion of p50 The expression of TF E selectin and VCAM 1 was also markedly upregulated in the patient sample of thrombotic vasculitis indicating the clinical relevance of NF κB activation in the pathogeneses of occlusive arterial diseases Our data thus indicate that by the downregulation of the NF κB target genes that are critical in thrombosis and inflammation specific inhibitors of p50 such as Andro may be therapeutically valuable for preventing and treating thrombotic arterial diseases including neointimal hyperplasia in arterial restenosis IL 4 protects the B cell lymphoma cell line CH31 from anti IgM induced growth arrest and apoptosis contribution of the PI 3 kinase AKT pathway Gregory B Carey 1 2 Elena Semenova 1 Xiulan Qi 1

    Original URL path: http://www.cell-research.com/artsmore.asp?id=68 (2016-02-14)
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