约氏疟原虫Pys48核酸疫苗传播阻断效应的实验研究

探讨约氏疟原虫（Plasmodium yoelii17XL）Pys48核酸疫苗免疫BALB/c小鼠的特异性抗体产生特点及其效应。将Pys48核酸疫苗肌肉内注射免疫BALB/c小鼠,并以空质粒注射组作为对照,3次免疫后通过P.y17XL攻击小鼠;采用ELISA检测免疫后小鼠血清中特异性抗体水平;通过P.y17XL感染小鼠,取其感染后第3天含有配子体的血液进行体外培养,观察合子、动合子的形成数量。ELISA结果显示疫苗免疫组小鼠血清特异性抗体滴度明显高于对照组;而合子、动合子数量明显低于对照组。提示Pys48核酸疫苗具有良好的免疫原性,免疫小鼠后可以建立起有效的传播阻断效应。     

微波辅助提取荷叶生物碱条件的优化

对荷叶中主要生理活性物质生物碱的提取条件进行了初步研究,以稀盐酸水为溶剂结合微波照射浸渍提取,获得了较优化的提取工艺,即用pH2.5 HCl、微波辅助处理2.5 min、固液比1:40、提取温度90℃、提取时间3 h,可得荷叶生物碱量172.63μg.g-1。     

Pyrosequencing Reveals Contrasting Soil Bacterial Diversity and Community Structure of Two Main Winter Wheat Cropping Systems in China

Microbes are key components of the soil environment, playing an important role in maintaining soil health, sustainability, and productivity. The composition and structure of soil bacterial communities were examined in winter wheat–rice (WR) and winter wheat–maize (WM) cropping systems derived from five locations in the Low-Middle Yangtze River plain and the Huang-Huai-Hai plain by pyrosequencing of the 16S ribosomal RNA gene amplicons. A total of 102,367 high quality sequences were used for multivariate statistical analysis and to test for correlation between community structure and environmental variables such as crop rotations, soil properties, and locations. The most abundant phyla across all soil samples were Proteobacteria, Acidobacteria, and Bacteroidetes. Similar patterns of bacterial diversity and community structure were observed within the same cropping systems, and a higher relative abundance of anaerobic bacteria was found in WR compared to WM cropping systems. Variance partitioning analysis revealed complex relationships between bacterial community and environmental variables. The effect of crop rotations was low but significant, and interactions among soil properties, locations, and crop rotations accounted for most of the explained variation in the structure of bacterial communities. Soil properties such as pH, available P, and available K showed higher correlations (positive or negative) with the majority of the abundant taxa. Bacterial diversity (the Shannon index) and richness (Chao1 and ACE) were higher under WR than WM cropping systems.     

荒漠植物白刺属4个物种的生殖分配比较

选定乌兰布和沙漠地区白刺属4种植物为研究对象,通过对其样株在花期的各生殖构件的数量特征及生物量调查,系统研究了唐古特白刺(Nitraria tangutorum Bobr.)、西伯利亚白刺(Nitraria sibirica Pall.)、大白刺(Nitraria roborowskii Kom.)和泡泡刺(Nitraria sphaerocarpa Maxim.)4种白刺属植物在生殖枝水平上的生殖分配。结果表明:不同白刺属植物在分株高度、生殖枝长、生殖枝基径、单枝花数、花序干重、枝叶干重等生殖构件的数量性状方面均有显著差异,其中泡泡刺的各生殖构件的数量均最小;除了西伯利亚白刺的生殖分配值达到44.51%外,其余3种白刺的生殖分配值均没有超过20%。经统计分析,4种白刺种群的生殖枝长分别与分株高度呈显著(P<0.05)的直线性正相关关系;生殖枝花序干重与分株高显著正相关;4种白刺的生殖分配随着分株生殖枝生物量的增加而减少,即白刺的个体大小与生殖分配之间呈现负相关关系。这种生殖分配特点反映了不同白刺植物对生长环境的资源利用、与克隆繁殖的权衡及对生态适应的策略。     

基于TM的渤海海岸带1988～2000年生态环境变化

海岸带作为海陆之间的过渡地带,是全球生态环境最为复杂和特殊之处。研究海岸带土地利用变化对于了解该区域生态环境演变具有重要意义。利用1988和2000年的Landsat-TM数据,在GIS技术支持下,通过一系列空间分析,得到渤海海岸带土地利用/土地覆盖变化,结合社会经济统计资料分析该区域生态环境的动态变化情况及其驱动因素。结果表明,1988~2000年,由于渤海海岸带社会经济的快速发展,海岸带土地利用格局发生了巨大的变化。耕地大面积减少,城乡工矿用地、养殖池塘、盐田急剧扩张;林地、湿地等具有重要生态价值的土地类型面积显著下降。表明强烈的人类活动已经使自然生态系统受到破坏,渤海海岸带生态环境质量总体上呈现下降趋势。     

反义CD151基因转染对大鼠血管平滑肌细胞迁移的影响

目的观察pcDNA3.1真核表达载体介导的反义CD151基因转染对培养的大鼠动脉平滑肌细胞(VSMCs)迁移的影响。方法构建携带全长正义和反义CD151的真核表达载体pcDNA3.1-CD151和pcDNA3.1-anti-CD151重组质粒,转染体外培养的VSMCs,以RT-PCR和Western blot方法检测CD151的表达,用Boyden趋化小室方法观察细胞迁移。结果与载体对照组、脂质体对照组和空白对照组3组均值比较,转染48h后,反义CD151组mRNA表达降低58%,蛋白表达降低51%,正义CD151组的VSMCs CD151mRNA表达增加171%,蛋白表达增加133%;趋化迁移的细胞数,反义CD151组为37.9±6.3,正义CD151组为86.5±12.4;载体对照组、脂质体对照组和空白对照组分别为60.3±7.1、61.8±7.6和67.3±9.6。反义CD151组显著低于其余各组(P<0.01),正义CD151组显著高于其余各组(P<0.01)。结论pcDNA3.1真核表达载体介导的反义CD151转染,通过抑制CD151的表达,能够显著抑制大鼠VSMCs的迁移。     

利用噬菌体随机肽库筛选可结合志贺毒素B亚单位的短肽序列

以制备的重组志贺毒素B亚单位(StxB)为靶标,利用噬菌体展示亲和淘选技术,经4轮筛选,从随机十二肽库中筛选到与StxB结合的一批噬菌体克隆,对特异结合活性较高的27个噬菌体克隆的表面展示肽进行序列测定,其中A6序列出现16次,A9和A3序列分别出现2次和3次。为评价筛选克隆中和毒素毒性的能力,将展示肽出现频率最高的A6噬菌体克隆,体外与志贺毒素孵育进行动物试验,动物存活率达33.3%,表明毒素的毒性得到部分抑制,A6短肽可能发展成为志贺毒素的拮抗剂。     

花生根瘤菌的抗逆性初步研究*

对３９株花生根瘤菌（其中６株为引进菌株）和９株参比菌株进行干旱、ＮａＣｌ、ｐＨ及低温的耐受性实验。结果表明我国花生根瘤菌资源中存在着耐盐、酸较强的菌株,且耐受性水平差异较大;在８℃条件下供试菌株未见生长;花生根瘤菌有着比大豆根瘤菌更强的抗干旱能力,抗干旱能力同耐盐能力之间没有明显的关系。另外,菌株的抗逆性同共生特性、分离地之间也未见有明显的关系。     

Regulation of cell-cell interactions by phosphatidic acid phosphatase 2b/VCIP

We identified vascular endothelial growth factor and type I collagen inducible protein (VCIP), also known as phosphatidic acid phosphatase 2b (PAP2b), in a functional assay of angiogenesis. VCIP/PAP2b exhibits an Arg-Gly-Asp (RGD) cell adhesion sequence. Immunoprecipitation and fluorescence-activated cell sorting analyses demonstrated that VCIP-RGD is exposed to the outside of the cell surface. Retroviral transduction of VCIP induced cell aggregation/cell- cell interactions, modestly increased p120 catenin expression and promoted activation of the Fak, Akt and GSK3beta protein kinases. Furthermore, expression of recombinant VCIP promoted adhesion, spreading and tyrosine phosphorylation of Fak, Shc, Cas and paxillin in endothelial cells. GST-VCIP-RGD, but not GST-VCIP-RGE, specifically interacted with a subset of integrins, and these interactions were effectively blocked by anti-alpha(v)beta(3) and anti-alpha(5)beta(1) integrin antibodies, and by PAP2b/VCIP-derived peptides. Interestingly, PAP2b/VCIP is expressed in close proximity to vascular endothelial growth factor, von Willebrand factor and alpha(v)beta(3) integrin in tumor vasculatures. These findings demonstrate an unexpected function of PAP2b/VCIP, and represent an important step towards understanding the molecular mechanisms by which PAP2b/VCIP-induced cell-cell interactions regulate specific intracellular signaling pathways.     

鲎源抗菌肽对大肠杆菌的抑杀机理

【目的】研究鲎源抗菌肽鲎素对大肠杆菌抑杀的作用机理,为防治大肠杆菌引起的肠道疾病提供新的潜在抗菌药物。【方法】利用牛津杯法和微量MH肉汤稀释法测定其抗菌活性,激光共聚焦扫描显微镜观察鲎素对大肠杆菌的结合、分布及杀伤过程,透射电镜观察鲎素对大肠杆菌超微结构的影响,并采用琼脂糖凝胶阻滞电泳研究其对大肠杆菌基因组DNA和RNA的影响。【结果】鲎素对大肠杆菌的最小抑菌浓度与最低杀菌浓度分别为5 mg/L和20 mg/L;激光共聚焦扫描显微镜和透射电镜观察发现鲎素能快速作用于细胞表面,并发生聚集现象,随着作用时间的延长能导致细胞膜结构的破坏和细胞内含物的释放;通过琼脂糖电泳结果显示鲎素也能够作用于大肠杆菌基因组DNA,并呈浓度依赖关系,10 mg/L鲎素对基因组DNA无明显影响,80 mg/L鲎素能导致DNA断裂;凝胶阻滞电泳显示鲎素也能与基因组DNA和RNA发生结合。【结论】研究结果为深入探讨鲎素抑杀大肠杆菌的分子机制提供重要的理论依据。