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染色质动态调整与基因转录

生物工程进展 1999年第4期19卷 23-27,10页

作者： Shen Yufei (national laboratory of medical molecular biology,institute of basic medical sciences chinese academy of medical sciences & peking union medical sciences beijing 100005,china) 中国医学科学院中国协和医科大学

核小体是真核细胞染色质的基本单位。真核基因组ＤＮＡ在细胞核中储存在染色质结构中。核小体的核心结构是由１４６个碱基对的ＤＮＡ盘绕四对核心组蛋白的八聚体外面近两周所构成。核小体核心组蛋白Ｈ３／Ｈ４两对异源二聚体是核小体亚... 详细信息

核小体是真核细胞染色质的基本单位。真核基因组ＤＮＡ在细胞核中储存在染色质结构中。核小体的核心结构是由１４６个碱基对的ＤＮＡ盘绕四对核心组蛋白的八聚体外面近两周所构成。核小体核心组蛋白Ｈ３／Ｈ４两对异源二聚体是核小体亚颗粒的核心，它能稳定地与ＤＮＡ结合...

关键词：染色质核小体动态调整基因转录

Protein expression profile in the differentiation of rat bone marrow stromal cells into Schwann cell-like cells

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science china(Life sciences) 2009年第3期52卷 267-277页

作者： LI WenTing1,SUN HuaLin2,XU ZengLu1,DING Fei2 & GU XiaoSong2 1 Department of Anatomy,Histology and Embryology,peking union medical college,institute of basic medical science,chinese academy of medical sciences,beijing 100005,china 2 Jiangsu Key laboratory of Neuroregeneration,Nantong University,Nantong 226001,china 1. Department of Anatomy Histology and Embryology Peking Union Medical College Institute of Basic Medical Science Chinese Academy of Medical Sciences Beijing 100005 China 2. Jiangsu Key laboratory of Neuroregeneration Nantong University Nantong 226001 China

During the last decade,increasing evidence suggested that bone marrow stromal cells(MSCs) have the potential to differentiate into neural *** studies have reported that MSCs showed morphological changes and expressed a limited number of neural proteins under experimental ***,no proteomic studies on MSCs differentiated into Schwann cell-like cells have been *** this study,we isolated MSCs from adult Sprague-Dawley rat femur and tibia bone marrows and induced the cells in vitro under specific *** using two-dimensional gel electrophoresis(2-DE),we compared the protein profiles of MSCs before and after induced *** obtained 792 protein spots in the protein profile by 2-DE,and found that 74 spots changed significantly before and after the differentiation using PDQuest software,with 43 up-regulated and 31 *** analyzed these 74 spots by a matrix assisted laser desorption ionization-time of flight mass spectrometry(MALDI-TOF-MS) and by database searching,and found that they could be grouped into various classes,including cytoskeleton and structure proteins,growth factors,metabolic proteins,chaperone proteins,receptor proteins,cell cycle proteins,calcium binding proteins,and other *** proteins also include neural and glial proteins,such as BDNF,CNTF and *** results may provide valuable proteomic information about the differentiation of MSCs into Schwann cell-like cells.

关键词： one marrow stromal cells(MSCs) differentiation Schwann cell-like cells proteomics

人ATP7B基因对Wilson病动物模型LEC大鼠暴发性肝炎的治疗

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解剖学报 2004年第3期35卷 282-286页

作者：孟雁苏牧中国医学科学院基础医学研究所中国协和医科大学基础医学院医学分子生物学国家重点实验室北京100005

目的　探讨人ATP7B基因对Wilson病动物模型LEC大鼠暴发性肝炎的治疗效果。　方法　将构建的 7.1kb含有鸡 β肌动蛋白启动子的人正常ATP7BcDNA ,经显微注射法导入人Wilson病动物模型LEC(long EvansCinnamon)大鼠受精卵 ,建立转基因功能... 详细信息

目的　探讨人ATP7B基因对Wilson病动物模型LEC大鼠暴发性肝炎的治疗效果。　方法　将构建的 7.1kb含有鸡 β肌动蛋白启动子的人正常ATP7BcDNA ,经显微注射法导入人Wilson病动物模型LEC(long EvansCinnamon)大鼠受精卵 ,建立转基因功能恢复大鼠模型。以Westernbolt检测人ATP7B在转基因大鼠肝内的表达 ,用同窝无转基因大鼠及正常野生型LEA大鼠作对照 ,对 6～ 16周龄的转基因大鼠的血清AST、ALT和总胆红素水平进行连续测定 ,同时取 13周龄转基因大鼠的肝组织进行病理学和组织化学分析。　结果　人ATP7B基因在转基因大鼠的肝组织中获得正确和完整的表达 :12周龄前后 ,与同窝无转基因大鼠相比 ,转基因大鼠的血清AST、ALT和总胆红素水平明显降低 ,肝组织的炎性病变显著减轻 ,肝细胞内铜的蓄积减少 ;至 16周龄 ,转基因大鼠的临床表型未见异常 ,存活率达 10 0 %。说明人ATP7B的导入 ,成功地抑制了Wilson病动物模型LEC大鼠的暴发性肝炎的发生。结论　Wilson病肝炎的发生与ATP7B基因功能缺陷引起的铜蓄积有直接关系。基因转移有可能是一种有效的治疗Wilson病的方法。

关键词：人ATP7B基因基因治疗暴发性肝炎 LEC大鼠

人ATP7B基因对Wilson病动物模型LEC大鼠肝硬化及肝癌的治疗

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解剖学报 2004年第4期35卷 405-408页

作者：孟雁苏牧中国医学科学院中国协和医科大学基础医学研究所医学分子生物学国家重点实验室北京100005

目的　探讨人ATP7B基因对Wilson病动物模型LEC(Long EvansCinnamon)大鼠肝硬化及肝癌的治疗效果。　方法　将 7 1kb含有鸡 β肌动蛋白启动子的人正常ATP7BcDNA ,经显微注射法导入Wilson病动物模型LEC大鼠受精卵 ,建立转基因功能恢复大... 详细信息

目的　探讨人ATP7B基因对Wilson病动物模型LEC(Long EvansCinnamon)大鼠肝硬化及肝癌的治疗效果。　方法　将 7 1kb含有鸡 β肌动蛋白启动子的人正常ATP7BcDNA ,经显微注射法导入Wilson病动物模型LEC大鼠受精卵 ,建立转基因功能恢复大鼠模型。以无转基因大鼠及正常野生型LEA大鼠为对照 ,对 17～ 30周龄的转基因大鼠的血清AST、ALT水平进行连续测定 ,同时取 30和 6 0周龄转基因大鼠的肝脏进行病理组织学和组织化学分析。　结果　 17～ 30周龄 ,转基因大鼠的血清丙氨酸氨基转移酶 (AST)、天门冬氨酸氨基转移酶 (ALT)保持在较低的水平。至 6 0周龄 ,雄性转基因大鼠肝组织未见胆管纤维化。在所有被检的转基因大鼠个体未见肝细胞癌性病变。转基因大鼠的存活率达 95 7%。此外 ,30和 6 0周龄的转基因大鼠肝细胞内铜着色颗粒的分布及数量无明显变化。　结论　人ATP7B的导入有效的延缓了Wilson病动物模型LEC大鼠肝硬化的形成、抑制了肝癌的发生。Wilson病的肝硬化及肝癌的发生可能与铜的蓄积无直接关系。

关键词：人ATP7B 基因治疗肝硬化肝癌 LEC大鼠

编码内质网膜蛋白的新基因家族RTN的研究进展

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生物化学与生物物理进展 2001年第1期28卷 40-43页

作者：黄晓玮阴彬袁建刚强伯勤中国医学科学院基础医学研究所中国协和医科大学基础医学院医学分子生物国家重点实验室北京100005 国家人类基因组北方研究中心北京100073

蛋白质的分泌渠道是近几年生命科学研究焦点之一 .RTN (reticulon)是定位于内质网膜上的一个基因家族 ,现已发现RTN1、RTN2、RTN3、RTN4等多个家族成员 .其中 ,这些基因在蛋白质分泌加工过程中的具体作用 ,以及RTN1(即NSP)作为神经内分... 详细信息

蛋白质的分泌渠道是近几年生命科学研究焦点之一 .RTN (reticulon)是定位于内质网膜上的一个基因家族 ,现已发现RTN1、RTN2、RTN3、RTN4等多个家族成员 .其中 ,这些基因在蛋白质分泌加工过程中的具体作用 ,以及RTN1(即NSP)作为神经内分泌特异性的标志或与神经元分化的关系都在进一步探讨之中 .

关键词：内质肉膜蛋白蛋白质分泌神经内分泌特异蛋白分泌渠道

Calcitonin gene-related peptide inhibits interleukin-1β-induced interleukin-8 secretion in human type Ⅱ alveolar epithelial cells

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Acta Pharmacologica Sinica 2006年第10期27卷 1340-1345页

作者： Wen-jing LI Teng-ke WANG Xian WANG~2 Department of Physiology,School of basic medical sciences,peking University,Key laboratory of molecular Cardiovascular science,Ministry of Education,beijing 100083,china Department of Physiology School of Basic Medical Sciences Peking University Key Laboratory of Molecular Cardiovascular Science Ministry of Education Beijing 100083 China

Aim:Our previous data have shown that type Ⅱ alveolar epithelial(AEⅡ)cellsexpress neuropeptide calcitonin gene-related peptide(CGRP),and that pro-in-flammatory factor interleukin 1-β(IL-1β)induces CGRP secretion in the A549 hu-man AEⅡ cell *** the present study,we investigated the effect of endogenousand exogenous CGRP on IL-1β-induced chemokine interleukin-8(IL-8)***:We used enzyme-linked immunosorbent assay(ELISA)and RT-PCR todetect IL-8 protein and mRNA levels,respectively,siRNA and the stably trans-fected cell line were used to knock down and overexpress the CGRP gene,respectively,and chemiluminescence assay was used to detect reactive oxygenspecies(ROS)***:CGRP-1 receptor antagonist hCGRP8-37(0.1-1nmol·L-1)greatly amplified IL-1β-induced IL-8 *** inhibition of CGRPexpression by siRNA significantly increased IL-8 secretion upon IL-1β ***,cell clones stably transfected with CGRP showed significantly inhibitedmRNA and protein levels of IL-8 induced by 1L-1β.Conclusion:These data implythat AEⅡ cell-derived CGRP suppress IL-1β-induced IL-8 secretion in an autocrine/paracrine *** investigation showed that CGRP attenuated IL-1β-arousedROS formation,which is an early indication of pro-inflammatory factor signaling.

关键词： type Ⅱ alveolar epithelial cells interleukinl interleukin-8 calcitonin gene-related peptide anti-inflammation lung reactive oxygen species

Chromatin domain boundaries: insulators and beyond

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Cell Research 2005年第4期15卷 292-300页

作者： Chih Chuan LIANG national laboratory of medical molecular biology institute of basic medical sciences chinese academy of medicalsciences and peking union medical college 5 Dong Dan San Tiao beijing 100005 china

The eukaryotic genome is organized into functionally and structurally distinct domains, representing regulatory unitsfor gene expression and chromosome behavior. DNA sequences that mark the border between adjacent domains are theinsulators or boundary elements, which are required in maintenance of the function of different domains. Some insula-tors need others enable to play insulation activity. Chromatin domains are defined by distinct sets of post-translationallymodified histones. Recent studies show that these histone modifications are also involved in establishment of sharpchromatin boundaries in order to prevent the spreading of distinct domains. Additionally, in some loci, the high-orderchromatin structures for long-range looping interactions also have boundary activities, suggesting a correlation betweeninsulators and chromatin loop domains. In this review, we will discuss recent progress in the field of chromatin domainboundaries.

关键词： insulator chromatin domain boundary histone code chromatin loop domain.

Recombinant Neural Protein PrP Can Bind with Both Recombinant and Native Apolipoprotein E In Vitro

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Acta Biochimica et Biophysica Sinica 2006年第9期38卷 593-601页

作者： Chen GAO~1 Yan-Jun LEI~(1,2) Jun HAN~1 Qi SHI~1 Lan CHEN~(1,3) Yan GUO~(1,4) Yong-Jun GAO~1 Jian-Ming CHEN~1 Hui-Ying JIANG~1 Wei ZHOU~1 Xiao-Ping DONG~1* 1 State Key laboratory for Infectious Disease Prevention and Control,national institute for Viral Disease Control and Prevention,chinese Center for Disease Control and Prevention,beijing 100052,china 2 School of Medicine,Xi'an Jiaotong University,Xi'an 710061,china 3 national laboratory of medical molecular biology,institute of basic medical science,chinese academy of medical sciences and peking union medical college,beijing 100005,china 4 college of science and Veterinary Medicine,Northwest Agriculture and Forest University,Yangling 712100,china StateKeylaboratoryforInfectiousDiseasePreventionandControl NationalInstituteforViralDiseaseControlandPreventionChineseCenterforDiseaseControlandPreventionBeijing100052China SchoolofMedicine Xi'anJiaotongUniversity

The most essential and crucial step during the pathogenesis of transmissible spongiformencephalopathy is the conformational change of cellular prion protein （PrPC） to pathologic isoform (PrPSc).Alot of data revealed that caveolae-like domains （CLDs） in the cell surface were the probable place where theconversion of PrP proteins *** E （ApoE） is an apolipoprotein which is considered toplay an important role in the development of Alzheimer's disease and other neurodegenerative diseases byforming protein complex through binding to the receptor located in the clathrin-coated pits of the cell *** this study,a 914-bp cDNA sequence encoding human ApoE3 was amplified from neuroblastoma cell *** human ApoE isomers were expressed and purified from Escherichia ***-specificantiserum was prepared by immunizing rabbits with the purified ***/His pull-down assay,immunoprecipitation and ELISA revealed that three full-length ApoE isomers interact with the recombinantfull-length PrP protein in *** regions corresponding to protein binding were mapped in the N-terminalsegment of ApoE （amino acid 1-194） and the N-terminal of PrP （amino acid 23-90）.Moreover,the recombinantPrP showed the ability to form a complex with the native ApoE from liver *** data provided directevidence of molecular interaction between ApoE and *** also supplied scientific clues for assessing thesignificance of CLDs on the surface of cellular membrane in the process of conformational conversion fromPrPCto PrPScand probing into the pathogenesis of transmissible spongiform encephalopathy.

关键词： prion disease PrP apolipoprotein E protein interaction caveolae-like domain

A competitive RT-PCR-based approach to quantifying promoter activity of stress gene

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Progress in Natural science:Materials International 2002年第10期 24-28页

作者： MO Zhicheng, LI Xiaoyan, LI Hongfan, CHENG Xiaokuan, WU Ninghua and SHEN Yufei(national laboratory of medical molecular biology, institute of basic medical sciences, chinese academy of medical sciences & peking union medical college, beijing 100005, china) national laboratory of medical molecular biology Institute of Basic Medical Sciences Chinese Academy of Medical Sciences & Peking Union Medical College Beijing 100005 China

Chloramphenicol acetyltransferase (CAT) as a stable reporter has been broadly adopted to detect promoter activity of many stress genes in eukaryotic cells. A simplified, convenient and relatively accurate assay to reveal CAT activities driven by heat shock gene promoter upon cell stress is reported here. A CAT mutant control plasmid pMCAT is constructed out of pBLCAT3, with a DNA segment within the CAT gene deleted. The control plasmid pMCAT along with the wild type CAT reporter gene driven by the regulatory sequence of hsp90β gene are co-transfected into Jurkat cells. Following total cellular RNA extracted and reverse transcribed, PCR is carried out to amplify two individual fragments with the same pair of synthesized primers, which bind to the identical sequences in both pMCAT and the CAT reporter plasmids. The two PCR fragments amplified from the reporter and control RNAs are identified by their size differences in agarose gel electropboresis. The ratio of intensity of the two PCR bands in the same

关键词： chloramphenicol acetyltransferase mRNA promoter activity competitive RT-PCR.