利用K562红细胞分化模型和cDNA芯片技术筛选参与红细胞分化的新基因(英文)

以氯高铁血红素 (hemin)诱导K5 6 2分化作为体外红细胞分化模型 ,结合cDNA大规模测序、生物信息学分析、基因芯片杂交和NorthernBlot分析等技术 ,筛选红细胞分化相关的新基因 .首先利用大规模测序技术从人胚肾cDNA文库中随机挑选克隆测得 192个EST(expressedsequencetags)片段 ,经在线生物信息学分析 ,得到 79个代表新基因的未知EST片段 ,并在NCBI(NationalCenterofBiotechnologyInformation)dbEST库中登录 .利用 79个ESTcDNA片段制备了基因芯片 .提取分化前后的K5 6 2细胞的mRNA作为荧光标记反转录的模板 ,反转录后的探针用于DNA芯片杂交 .分析杂交后的结果 ,得到了 2个差异表达较明显的基因 ,GenBank登录号分别为AF147772 (187bp)和AF4 776 2(6 30bp) ,并分别命名为EDRG1和EDRG2 (erythroiddifferentiationrelatedgene 1and 2 ) ,相似性检索表明它们属全新基因 ,基因组草图测序数据库检索表明了两个基因的染色体定位 .随后的Northern印迹用于验证了在分化前后的K5 6 2细胞中差异表达 .提示这两个基因参与了红细胞分化过程 .RT PCR检测了EDRG1和EDRG2在人胚胎多组织中的表达 .结果提示 ,EDRG1可能与多种胚组织的正常发育相关 ,尤其在胚脑中高丰度表达 ,而EDRG2则可能参与了胚心和胚肾的组织生成 .生物     

王锦蛇的人工养殖方法

王锦蛇是一种比较适合人工养殖的无毒蛇。王锦蛇体大、耐寒、适应性强、生长快、饲养周期短、易孵化 ,很多蛇场或养蛇户多以王锦蛇作为蛇类的养殖品种 ;王锦蛇特别适合在我国北方各省区养殖。作者以北方地区为例 ,浅述王锦蛇的养殖方法。1　王锦蛇的生长习性　　王锦蛇又名菜花蛇、大王蛇、臭薰领等 ,无毒 ,常栖息在山地、平原及丘陵地带 ,活动于河边、水塘边、库区及其它近水域的地方。王锦蛇动作敏捷、性情凶猛、爬行速度快且会攀树 ,属广食性蛇 ,常以蛙类、鸟类、鼠类及各种鸟蛋为食。食物缺乏时 ,它甚至吞食自己的幼蛇和同类。2　王锦蛇…     

Initial function analysis of a novel erythroid differentiation related gene EDRF1

Erythroid differentiation depends on the establishment of specific patterns of gene expression. Hypersensitive site 2 (HS2, serving as a major enhancer of globin genes)-binding proteins may be involved in its natural open chromosomal environment formation. Previously we prepared monoclonal antibodies against HS2-binding nuclear proteins of terminal differentiated erythroid cells. By utilizing the monoclonal antibodies, we screened λ-gt11 human fetal liver cDNA expression library and obtained one cDNA clone, which was named erythroid differentiation related gene (EDRF1, Genbank accession number AF040247) , encompassing an entire open reading frame. We investigated the expression pattern of EDRF1 by RT-PCR technique. And a clue to the function of EDRF1 has been found from confirmation of high levels of EDRF1 mRNA in differentiated K562 and human fetal liver tissue. To illuminate the function of EDRF1 in K562 cells, sense and antisense EDRF1 constructs were prepared and transfected into K562 cells, α-glob     

肝脏星形细胞的生物学特性研究进展

肝脏星形细胞(HSCs)是肝脏的非实质细胞之一,具有重要的生理功能,在病理情况下,HSCs被激活,具有许多重要的生物学特性,在肝纤维化与门静脉高压的形成中发挥重要作用.对影响HSCs激活的因素、HSCs激活及凋亡机制的研究成为近年来肝脏细胞学研究的热点之一,并取得了很大进展.     

Copia 类型反转录转座子在籽粒苋中的表现

利用籽粒苋为材料 ,通过PCR扩增、克隆研究了copia类型反转录转座子的反转录酶序列在籽粒苋中的表现 ,结果表明 :(1)反转录转座子在苋属的 30个品系中同时检测到 ,说明反转录转座子在籽粒苋的不同种中普遍存在 ;(2 )对野生苋 (A .quitensis)中克隆、序列分析 2 8个反转录转座子的反转录酶序列 ,碱基分析表明 ,它们存在高度异质性 ,不同的反转录酶序列均存在不同程度的碱基缺失突变和终止密码子的突变 ;(3)对 2 8个序列与已发表的一些不同物种 (如水稻、矮牵牛和果蝇等 )的同一类型序列聚类分析表明 ,存在于野生苋中的反转录转座子与来源于双子叶或单子叶植物的反转录转座子关系密切 ,它们与果蝇的copia因子和 1731因子可能具有相同的起源     

Antisense EDRF1 gene inhibited GATA-1 transcription factor DNA-binding activity in K562 cells

Our previous studies showed that EDRF1 influenced expression of a-globin mRNA and synthesis of hemoglobin in K562 cells and modulated self-renewal of K562 cells. To illuminate the function of EDRF1 in K562 cells, sense and antisense EDRF1 constructs were prepared and transfected into K562 cells. By using microarray and dot blot assay, 60 cytokine receptors and some oncogenes sharing important functions in cell proliferation and differentiation were investigated. The results of this study demonstrated that IL-6 receptor, GM-CSF receptor, c-Jun/c-Fos, c-Myc and c-kit genes were regulated by antisense EDRF1 expression. The regulation was confirmed by RNA blot assay. GATA-1 mRNA expression was modulated by EDRF1 gene transfection. Electrophoretic mobility shift assay suggested that the DNA-binding activity of GATA-1 was remarkably inhibited in K562 cells expressing EDRF1 antisense gene. DNA binding activity of NF-E2 was at the same level as control experiment. Therefore EDRF1 may play a role in erythroid pro     

Initial hydrodynamic study on a new intraaortic axial flow pump: Dynamic aortic valve

Rotary blood pumps have been researched as implantable ventricular assist devices for years. To further reduce the complex of implanted axial pumps, the authors proposed a new concept of intraaortic axial pump, termed previously as "dynamic aortic valve (DAV)". Instead of being driven by an intraaortic micro-electric motor, it was powered by a magnetic field from outside of body. To ensure the perfusion of coronary artery, the axial flow pump is to be implanted in the position of aortic valve. It could serve as either a blood pump or a mechanical valve depending on the power input. This research tested the feasibility of the new concept in model study. A column, made from permanent magnet, is jointed to an impeller in a concentric way to form a "rotor-impeller". Supported by a hanging shaft cantilevered in the center of a rigid cage, the rotor-impeller can be turned by the magnetic field in the surrounding space. In the present prototype, the rotor is 8 mm in diameter and 15 mm in length, the impeller ha     

大蟾蜍冬眠前后血清中某些激素含量变化及其与糖代谢的关系

本文分析了大蟾蜍冬眠前，冬眠期和冬眠后三个不同时期血清皮质醇，血清Ｔ４和血清Ｔ３含量的变经并与同期血糖水平和肝糖元含量进行比较，结果表明：血清皮质醇在冬眠期呈显著下降趋势，冬眠后又明显回升，血糖浓度的变化也表现出这种；血清Ｔ４在冬眠期呈明显上升的趋势，冬眠后又降回皮冬眠前的水平上；血清Ｔ３一直保持上升趋势；肝糖元含一则一直保持大幅度下降的趋势。此外，本文不上述变化的生理学意义进行了讨论。     

靶向人白介素-6蛋白的治疗性单克隆抗体研究进展*

白介素-6(interleukin-6,IL-6)作为一种多效的细胞因子,参与机体内众多生理与病理过程。研究表明,IL-6首先与自身受体(IL-6R、gp130)形成异源六聚体复合物,进而激活下游信号转导通路,最终发挥生物学功能。 IL-6信号通路异常活化及功能失调与多种疾病密切相关,如自身免疫疾病、慢性炎症、恶性肿瘤等。另外IL-6的异常表达在新型冠状病毒肺炎(COVID-19)细胞因子风暴综合征(CSS)中也扮演重要角色。一般而言,阻断IL-6信号通路上的各关键节点,均可用于IL-6相关疾病的治疗。有别于阻断IL-6R或gp130等公共受体分子,阻断IL-6蛋白的治疗性单克隆抗体特异性更高,在临床研究中,部分品种显示出其独有的治疗特点及有益的疗效。现阶段只有1个靶向IL-6蛋白的单克隆抗体药物获美国FDA批准上市,以及超过8个治疗性单克隆抗体在临床研究阶段。重点对国内外靶向人IL-6蛋白的治疗性单克隆抗体及其临床应用进行综述。     

基于微卫星标记的鼋亲子鉴定技术

为研究鼋(Pelochelys cantorii)保种群体的遗传信息,利用鼋转录组文库, 筛选出了10对微卫星引物, 以此建立了基于 2 组各含5个微卫星位点多重 PCR 体系的鼋亲子鉴定技术, 并应用 2 组微卫星多重 PCR 体系, 通过 ABI3130 遗传分析仪及 PopGene3.2和CERVUS软件对56只子一代鼋进行个体基因型检测和群体遗传多样性分析, 结果显示, 鼋子一代群体的平均等位基因数为2.3, 微卫星位点的平均多态信息含量为0.3829, 平均观测杂合度和期望杂合度分别为0.6305和0.4767, 处中高度水平。对子代进行亲子分析, 10个微卫星位点的累积排除概率分别为73.14%(NE-1P)、90.98%(NE-2P)和98%(NE-3P), 能够满足亲子鉴定的需要。且4只亲本在生殖选择中表现出差异。鼋多重 PCR 亲子鉴定技术的建立为群体遗传多样性分析、家系鉴定管理和选择育种提供了有效的技术手段。