Specific heteromeric association of four transmembrane peptides derived from platelet glycoprotein Ib-IX complex

As the receptor on the platelet surface for von Willebrand factor, glycoprotein (GP) Ib-IX complex is critically involved in hemostasis and thrombosis. How the complex is assembled from GP Ibα, GP Ibβ and GP IX subunits, all of which are type I transmembrane proteins, is not entirely clear. Genetic and mutational analyses have identified the transmembrane (TM) domains of these subunits as active participants in assembly of the complex. In this study, peptides containing the transmembrane domain of each subunit have been produced and their interaction with one another characterized. Only the Ibβ TM sequence, but not the Ibα and IX counterparts, can form homo-oligomers in SDS-PAGE and TOXCAT assays. Following up on our earlier observation that a Ibβ-Ibα-Ibβ peptide complex (αβ₂) linked through native juxtamembrane disulfide bonds could be produced from isolated Ibα and Ibβ TM peptides in detergent micelles, we show here that addition of the IX TM peptide facilitates formation of the native αβ₂ complex, reproducing the same effect by the IX subunit in cells expressing the GP Ib-IX complex. Specific fluorescence resonance energy transfer was observed between donor-labeled αβ₂ peptide complex and acceptor-conjugated IX TM peptide in micelles. Finally, the mutation D135K in the IX TM peptide could hamper both the formation of the αβ₂ complex and the energy transfer, consistent with its reported effect in the full-length complex. Overall, our results have demonstrated directly the native-like heteromeric interaction among the isolated Ibα, Ibβ and IX TM peptides, which provides support for the four-helix bundle model of the TM domains in the GP Ib-IX complex and paves the way for further structural analysis. The methods developed in this study may be applicable to other studies of heteromeric interaction among multiple TM helices.     

双歧杆菌及其完整肽聚糖对脐血来源树突状细胞分泌IL-12的影响

目的探讨两歧双歧杆菌和不同剂量双歧杆菌的完整肽聚糖（WPG）对脐血来源树突状细胞（DC）分泌IL-12的影响。方法以双歧杆菌全菌（量）和不同剂量双歧杆菌WPG（1-8μg/ml）与脐血来源树突状细胞共培养,用ELISA的方法测定培养上清中IL-12的量。结果双歧杆菌和其WPG（1-6μg/ml）与树突状细胞共培养后,树突状细胞分泌的IL-12的量显著高于阴性对照组（P〈0.01）,当WPG量为1-5μg/ml时树突状细胞分泌的IL-12的量呈剂量依赖性,其中WPG量为5μg/ml时作用最为显著,WPG量为6μg/ml时分泌IL-12量减少,WPG量为8μg/ml时,分泌的IL-12量与阴性对照组差异无显著性（P〉0.05）。结论两歧双歧杆菌及其WPG能够刺激脐血来源的树突状细胞IL-12分泌;双歧杆菌WPG的免疫刺激作用呈一定的量效关系     

饰纹姬蛙7个Dmrt基因DM结构域的克隆及序列分析

采用简并PCR技术扩增了饰纹姬蛙(Microhyla ornata)Dmrt基因的DM结构域,经分子克隆、SSCP和DNA序列分析,获得了该基因家族的7个成员并分别命名为MoDmrtla、MoDmrtlb、MoDmrt3al、MoDmrt3a2、MoDmrt3a3、MoDmrt3b和MoDmrt5.与不同进化地位其他动物相关的Dmrt基因进行比对和聚类分析,结果显示该基因在系统进化上具有高度的保守性.     

芜菁原生质体的分离及绿色荧光蛋白的瞬时表达

目的：分离芜菁叶片原生质体,建立蛋白质在芜菁原生质体的瞬时表达系统。方法：以津田芜菁成叶为试材,酶解分离原生质体;通过PEG介导的转化,将编码绿色荧光蛋白（GFP）的瞬时表达载体转入原生质体中,用激光扫描共聚焦显微镜检测原生质体中GFP的表达情况。结果：分离出大量的津田芜菁原生质体,并获得了较高的转化效率,GFP在整个原生质体中都有表达。结论：建立了津田芜菁原生质体瞬时表达系统。     

线艺建兰的组织培养和快速繁殖(简报)

以线艺建兰茎尖、腋芽为外植体,接种在1/2MS 6-BA 3.0mg/L NAA 0.3mg/L培养基上形成原球茎,原球茎在1/2MS 6-BA 0.1mg/L NAA 1.0mg/L培养基上可大量增殖,并伸长生长形成丛生型根状茎;根状茎在MS 6-BA 2.0mg/L PP333 1.0mg/L NAA 0.5mg/L培养基上可分化成小苗,分化率达46.3%;小苗在MS IBA 1.0mg/L GA 0.5mg/L 香蕉泥100g/L的培养基上生根壮苗,生根率达100%.     

不同叶色红栌叶片中色素含量、酶活性及内含物差异的研究

为明确秋季不同叶色美国红栌叶片的生理差异,以秋季同一植株上红色、中间色、绿色三种颜色的美国红栌叶片为试材,测定了叶片中色素物质含量、酶活性以及叶片可溶性内含物的含量,结果表明：在红色叶片中,叶绿素含量较低,PAL、POD酶活性较高,花青素苷/叶绿素的比值较大,从而使叶色显现红色;而在绿色叶片中,叶绿素含量较高,PAL、POD酶活性较小,花青素苷/叶绿素的比值较低,叶片显现绿色。通过可溶性内含物测定可知,在红色叶片中的可溶性糖和蛋白质含量相对较高,均与花青素苷/叶绿素的比值达到显著相关水平,表明这些内含物的积累有利于花色素苷的合成。     

vWF与IL-6预测肾性高血压大鼠经历人工寒潮发生卒中的研究

目的:观察易卒中型肾血管性高血压大鼠(RHRSP)在经历人工寒潮中风前血液中vWF与IL-6含量变化从而预测中风的发生.方法:双肾双夹法制作RHRSP模型,经历人工寒潮并于寒潮前取血,酶联免疫吸附测定(ELISA)法检测vWF与IL-6含量.结果:脑梗塞组寒潮前vWF的表达明显高于非卒中组,脑梗死组和脑出血组寒潮前的IL-6含量明显高于非卒中组.结论:高血压患者血液中IL-6和vWF的含量可能预测经历寒潮时发生脑卒中的情况.     

Comparison of the Immunogenicities of HIV-1 Mutants Based on Structural Modification of env

Eleven env mutants were designed and generated by site-directed mutagenesis of the regions around Nab epitopes and deletions of variable regions in env.The immunogenicities of the generated mutants were evaluated using single-cycle infection neutralization assays with two pseudoviruses and IFN-γELISPOT.Overall,five mutants(dWt,M2,M5-2,M5-1 and dM7)induced highed neutralization activities for both pseudoviruses than plasmid Wt,while only two of the mutants(dWt and M5-2)showed significant differences(P<0.05).Two mutants(M2 and dM2)induced more Env-specific T cells than plasmid Wt.Statistically however,significance was only reached for mutant M2.Thus,properly modified HIV-1 Env may have the potential to induce potent cellular and humoral immune responses.     

模拟酸雨对小黑麦种子萌发及幼苗生理特性的影响

以小黑麦品种中饲237为材料。在不同pH值的模拟酸雨条件下对种子萌发及幼苗生理特性进行了研究。分别测定了发芽率、发芽势、发芽指数、活力指数和种子脱氢酶含量、幼苗鲜重、幼苗叶片叶绿素含量、叶片电导率等指标。结果表明,种子的各项发芽指标及种子脱氢酶含量、幼苗鲜重、叶绿素含量等随酸雨pH值减小整体呈降低趋势,叶片电导率则随酸雨pH值减小而有增加趋势。     

茶叶多糖的抗氧化活性研究

从婺源粗老绿茶叶中提取纯化出了茶叶多糖,研究了它对超氧自由基、DPPH自由基的清除作用,以及对β-胡萝卜素-亚油酸氧化体系的抑制作用,以此来评价该多糖的抗氧化活性.结果表明,茶叶多糖对超氧自由基、DPPH自由基有较好的清除作用,而对β-胡萝卜素-亚油酸氧化体系也有较明显的抑制作用.