细胞质膜蛋白质组学研究技术进展

质膜蛋白在细胞中执行着非常重要的功能。随着蛋白质组学的发展,细胞质膜蛋白质组学成为蛋白质组学研究的重要组成部分,它为质膜蛋白的生物功能研究及药物靶标的发现提供了新的途径。然而,质膜蛋白丰度低、疏水性强,对现有蛋白质组学研究技术提出了挑战。简要综述了近年来质膜蛋白质组研究的相关技术进展,包括富集、提取分离鉴定方法及定量和生物信息学研究方法等。     

静态单轴拉伸应变刺激对细胞有丝分裂方向的影响

力学刺激对细胞发育具有重要意义,它如何对细胞分化及组织形态的发生产生影响是一个尚未完全阐明的问题.细胞的有丝分裂过程与细胞增殖、分化以及胚胎发育、组织器官形态形成和损伤组织的修复再生等特性密切相关,例如,细胞的有丝分裂方向就是影响细胞极性分化,乃至组织形态发生的因素之一.那么,力学刺激是否通过改变细胞有丝分裂方向从而影响细胞的分裂分化呢?以小鼠成骨细胞系MC3T3为模型,探讨了静态单轴拉伸应变刺激对细胞形态、应力纤维排布方向和有丝分裂方向的影响.结果显示,在4%及8%静态单轴拉伸应变条件下,48 h之内细胞形态发生明显变化,细胞呈梭状,长轴沿应变方向排列,细胞骨架微丝呈束状平行排列,方向与应变方向相关.统计学分析表明,4%应变刺激48 h后、8%应变6 h后、8%应变12 b后、8%应变24 h后,及8%应变48 h后,分别有49%,43%,54%,54%,和62%的细胞应力纤维排列方向与单轴拉伸应变方向的夹角在300以内,以及50%,48%,56%,53%和62%的细胞有丝分裂方向与单轴应变方向夹角在300以内.统计学分析表明,细胞形态、应力纤维排布及有丝分裂方向与拉伸方向相关,且应力纤维排列方向和有丝分裂方向之间呈现高的相关性,这种相关性在拉伸刺激48 h后表现很明显,由此推测,存在力学刺激影响细胞形态及细胞应力纤维排布方向,控制有丝分裂方向的机制.     

陡脉冲电场对恶性肿瘤细胞杀伤效应的研究

观察不同参数（频率、脉宽、峰值）组合的陡脉冲电场对恶性肿瘤细胞的杀伤效应,将癌细胞悬液3ml盛于玻璃瓶中,每次施加不同参数组合的电脉冲20min,用MTT法测细胞死亡率,并制作生长曲线;观察细胞的超微结构变化。结果发现：不同参数组合的陡脉冲对癌细胞的杀伤有较大影响,随着参数值的增大,细胞死亡率上升,细胞的增殖受到抑制;实验组的细胞肿胀,溶解性坏死,仅留细胞轮廓,细胞膜完整性破坏,线粒体空化、肿胀。     

中国版纳小型猪近交系动脉异种移植靶抗原研究

目的　研究中国版纳小型猪近交系动脉的异种靶抗原α -Gal的分布和半定量分析 ,为研究异种移植超急性排斥反应和异种生物材料提供资料。方法　通过亲和免疫组织化学法和图象分析对 10头版纳小型猪的四级动脉进行α Gal的分布和半定量研究。结果　 (1)各级血管组织中血管内皮细胞阳性表达明显 ,弹性纤维、胶原纤维和平滑肌细胞未见表达 ,外膜有微弱表达 ;(2 )图象分析显示 ,管径越细 ,其内皮细胞的表达越高 (P <0 .0 1) ,各级动脉内皮细胞阳性表达的面积密度比的组间差异显著 (P <0 .0 1) ,t检验发现每两组间的动脉内皮细胞阳性表达面积密度比相差显著 (P <0 .0 1)。结论　 (1)版纳小型猪动脉内皮细胞均有异种抗原α Gal分布 ,但是分布不均匀 ;(2 )微血管血栓是异种移植超急性排斥反应中的重要环节 ,小型猪的异种器官移植、彻底解决异种移植超急性排斥反应的关键是防治微血管栓塞     

A designed system for assessing how sequence affects alpha to beta conformational transitions in proteins.

The role of amino acid sequence in conformational switching observed in prions and proteins associated with amyloid diseases is not well understood. To study alpha to beta conformational transitions, we designed a series of peptides with structural duality; namely, peptides with sequence features of both an alpha-helical leucine zipper and a beta-hairpin. The parent peptide, Template-alpha, was designed to be a canonical leucine-zipper motif and was confirmed as such using circular dichroism spectroscopy and analytical ultracentrifugation. To introduce beta-structure character into the peptide, glutamine residues at sites away from the leucine-zipper dimer interface were replaced by threonine to give Template-alphaT. Unlike the parent peptide, Template-alphaT underwent a heat-inducible switch to beta-structure, which reversibly formed gels containing amyloid-like fibrils. In contrast to certain other natural proteins where destabilization of the native states facilitate transitions to amyloid, destabilization of the leucine-zipper form of Template-alphaT did not promote a transformation. Cross-linking the termini of the peptides compatible with the alternative beta-hairpin design, however, did promote the change. Furthermore, despite screening various conditions, only the internally cross-linked form of the parent, Template-alpha, peptide formed amyloid-like fibrils. These findings demonstrate that, in addition to general properties of the polypeptide backbone, specific residue placements that favor beta-structure promote amyloid formation.     

H18杂交瘤抗凋亡能力的改造

利用PCR从pGEMTbcl-X_L质粒中获得bcl-X_L基因,构建真核表达载体pEF-bcl-X^{L,脂质体法转染杂交瘤细胞,G418筛选稳定表达株,Western blotting检测目的蛋白表达,流式细胞仪检测Bcl-X_L提高杂交瘤抗正丁酸钠诱导凋亡的功能。 将构建的编码鼠bcl-X_L基因的真核表达载体pEF-bcl-X_L,转染H18细胞后,获得稳定的表达株细胞;稳定表达Bcl-X_L的细胞具有抗正丁酸钠诱导凋亡的功能。鼠bcl-X_L基因在杂交瘤细胞中稳定表达,提高了杂交瘤抗凋亡的能力,对高密度大规模培养杂交瘤细胞具有重要意义。}     

Integrating a global agro-climatic classification with bioregional boundaries in Australia

Aim Stratification of major differences in the biophysical features of landscapes at the continental scale is necessary to collectively assess local observations of landscape response to management actions for consistency and difference. Such a stratification is an important step in the development of generalizations concerning how landscapes respond to different management regimes. As part of the development of a comparative framework for this purpose, we propose a climate classification adapted from an existing broad scale global agro‐climatic classification, which is closely aligned with natural vegetation formations and common land uses across Australia. Location The project considered landscapes across the continent of Australia. Methods The global agro‐climatic classification was adapted by using elevation‐dependent thin plate smoothing splines to clarify the spatial extents of the 18 global classes found in Australia. The clarified class boundaries were interpolated from known classes at 822 points across Australia. These classes were then aligned with the existing bioregional classification, Interim Biogeographic Regionalization for Australia IBRA 5.1. Results The aligned climate classes reflect major patterns in plant growth temperature and moisture indices and seasonality. These in turn reflect broad differences in cropping and other land use characteristics. Fifty‐two of the 85 bioregions were classified entirely into one of the 18 agro‐climatic classes. The remaining bioregions were classified according to sub‐bioregional boundaries. A small number of these sub‐bioregions were split to better reflect agro‐climatic boundaries. Main conclusions The agro‐climatic classification provided an explicit global context for the analysis. The topographic dependence of the revised climate class boundaries clarified the spatial extents of poorly sampled highland classes and facilitated the alignment of these classes with the bioregional classification. This also made the classification amenable to explicit application. The bioregional and subregional boundaries reflect discontinuities in biophysical features. These permit the integrated classification to reflect major potential differences in landscape function and response to management. The refined agro‐climatic classification and its integration with the IBRA bioregions are both available for general use and assessment.     

硫灯和氙灯下生长的黄瓜幼苗植物激素水平的差异

就人工气候室两种光源硫灯和氙灯引起黄瓜幼苗生长差异的原因进行了初步探讨.与生长在氙灯下的幼苗相比,硫灯下生长的幼苗较矮.组织细胞观察显示,硫灯下生长的黄瓜幼苗中胚轴表皮和皮层细胞纵向单位毫米长度所含细胞数目比氙灯下多.内源激素分析结果表明,硫灯下生长的黄瓜幼苗内源激素吲哚乙酸(IAA)和赤霉素(GA3)含量较氙灯下分别低17%和24%,而脱落酸(ABA)含量高31%.推测硫灯和氙灯下黄瓜幼苗生长的差异可能与内源激素调控有关.     

Substituted 2,2-bisaryl-bicycloheptanes as novel and potent inhibitors of 5-lipoxygenase activating protein

The discovery and SAR of a novel series of substituted 2,2-bisaryl-bicycloheptane inhibitors of 5-lipoxygenase activating protein (FLAP) are herein described. SAR studies have shown that 2,5-substitution on the exo-aryl group is optimal for potency. The most potent compounds in this series have an ortho-nitrogen aryl linked with a methyleneoxy as the 5-substituent and a polar group such as a urethane as the 2-substituent. One of the most potent compounds identified is the 5-benzothiazolymethoxy-2-pyridinylcarbamate derivative 2 (FLAP IC(50)=2.8 nM) which blocks 89% of ragweed induced urinary LTE(4) production in dogs (at an I.V. dose of 2.5 microg/kg/min). This compound inhibits calcium ionophore stimulated LTB(4) production in both human polymorphonuclear (PMN) leukocytes and human whole blood (IC(50)=2.0 and 33 nM, respectively).