中国东部沿海水仙归化群体的遗传多样性

为揭示我国东部归化水仙(Narcissus tazetta var. chinensis)的群体遗传多样性,利用2个叶绿体基因mat K和trn H-psb A片段对采自沪、浙、闽的5个代表群体的49株水仙进行了评估。结果表明,双基因联合序列的总长为1443bp,共定义6个单倍型,各归化群体的遗传多样性水平为DLSYPTDNJD=ZZCMD。AMOVA分析表明,群体内变异为遗传变异的主要来源(91.98%),群体间的遗传分化较低(Fst=0.080 22)。群体物种水平上的谱系结构不显著(Nst=0.020Gst=0.031;P0.05)。Mantel检验表明水仙群体间的遗传距离与地理距离呈显著的线性相关(r=0.929,P=0.02 0.05)。中性检验和错配分布分析结果均暗示水仙群体背离了快速扩张模型的假设。单倍型分布的中介网络图结合系统发育NJ树均将所有群体划分为2大分支。因此,我国东部沿海水仙归化群体整体遗传多样性水平较低,各群体间遗传分化较弱,遗传变异主要来自群体内,物种近期未经历扩张事件,可能是基因流受海岛隔离、自身生物学特征、生境异质性与及人为干扰的综合作用影响。     

microRNA在结核分枝杆菌抗细胞自噬作用中的研究进展

结核分枝杆菌(Mycobacteria tuberculosis,MTB)是结核病的致病菌,其感染机体后能在细胞内长期生存并在合适的条件下引发疾病.非氧依赖性杀伤是巨噬细胞清除MTB的重要途径,主要表现为细胞内吞体和溶酶体融合,利用细胞自噬作用清除内部细菌.相应的,MTB可利用多种方式顽强抵抗细胞自噬作用,与细胞共存从而逃避宿主免疫杀伤作用. MicroRNA(miRNA)是一种内源性非编码单链小RNA分子,其能在转录后水平沉默相关基因表达,是介导MTB与炎性细胞许多反应的重要分子.近期研究发现,MTB能够通过诱导巨噬细胞特异表达一些miRNA分子并靶向自噬相关基因,阻碍自噬发生、发展,从而实现MTB的抗细胞自噬作用.本文就miRNA在MTB抗细胞自噬中的作用及机制的研究进展作一综述.     

人二氢乳清酸脱氢酶蛋白的表达、纯化及其与新型抑制剂的晶体结构

人二氢乳清酸脱氢酶（human dihydroorotate dehydrogenase, hDHODH）是催化嘧啶从头合成途径的一个关键酶。近年来,多种研究表明,抑制该酶可缓解类风湿性关节炎的症状。但该酶的抑制剂甚少,寻找该酶的高效抑制剂具有重要意义。本研究利用PCR技术扩增hDHODH基因,构建重组质粒pET-19b-hDHODH,并在大肠杆菌(Escherichia coli, E.coli ) BL21(DE3)中表达,获得可溶性蛋白质。用Ni2+-NTA亲和层析柱对蛋白质进行纯化,获得较高（90%）纯度的hDHODH蛋白,将蛋白质与抑制剂3-(5-乙硫基)-1H-1, 2, 4-三氮唑-3-)苯甲酸和底物DHO混合孵育。用Hampton试剂盒初筛晶体并用棋盘法进行优化,获得晶形完美、衍射能力很强的hDHODH蛋白复合物单晶。用X射线衍射晶体,用CCP4、Coot软件解析结构,获得hDHODH蛋白复合物晶体结构。从解析的结构中可以看出,抑制剂与蛋白质的吻合度非常高,且抑制剂通过亲水的羧基端与蛋白质356位和147位的酪氨酸形成氢键网络。抑制剂的5元环与蛋白质359位的亮氨酸和360位的苏氨酸相互作用,使抑制剂与蛋白质牢固结合。该复合物晶体结构的顺利解析,将为开发新型特异性抗类风湿性关节炎药物提供重要基础。     

Dynamic evaluation of blood flow microcirculation by combined use of the laser Doppler flowmetry and high‐speed videocapillaroscopy methods

The dynamic light scattering methods are widely used in biomedical diagnostics involving evaluation of blood flow. However, there exist some difficulties in quantitative interpretation of backscattered light signals from the viewpoint of diagnostic information. This study considers the application of the high‐speed videocapillaroscopy (VCS) method that provides the direct measurement of the red blood cells (RBCs) velocity into a capillary. The VCS signal presents true oscillation nature of backscattered light caused by moving RBCs. Thus, the VCS signal can be assigned as a reference one with respect to more complicated signals like in laser Doppler flowmetry (LDF). An essential correlation between blood flow velocity oscillations in a separate human capillary and the integral perfusion estimate obtained by the LDF method has been found. The observation of blood flow by the VCS method during upper arm occlusion has shown emergence of the reverse blood flow effect in capillaries that corresponds to the biological zero signal in the LDF. The reverse blood flow effect has to be taken into account in interpretation of LDF signals.      

Mouse models of the laminopathies

The A and B type lamins are nuclear intermediate filament proteins that comprise the bulk of the nuclear lamina, a thin proteinaceous structure underlying the inner nuclear membrane. The A type lamins are encoded by the lamin A gene (LMNA). Mutations in this gene have been linked to at least nine diseases, including the progeroid diseases Hutchinson-Gilford progeria and atypical Werner's syndromes, striated muscle diseases including muscular dystrophies and dilated cardiomyopathies, lipodystrophies affecting adipose tissue deposition, diseases affecting skeletal development, and a peripheral neuropathy. To understand how different diseases arise from different mutations in the same gene, mouse lines carrying some of the same mutations found in the human diseases have been established. We, and others have generated mice with different mutations that result in progeria, muscular dystrophy, and dilated cardiomyopathy. To further our understanding of the functions of the lamins, we also created mice lacking lamin B1, as well as mice expressing only one of the A type lamins. These mouse lines are providing insights into the functions of the lamina and how changes to the lamina affect the mechanical integrity of the nucleus as well as signaling pathways that, when disrupted, may contribute to the disease.     

Application of cellulose-based self-assembled tri-enzyme system in a pseudo-reagent-less biosensor for biogenic catecholamine detection

Amorphous cellulose was used as a specific carrier for the deposition of self-assembled multienzyme complexes capable of catalyzing coupled reactions. Naturally glycosylated fungal cellobiohydrolases (CBHs) of glycosyl hydrolase families 6 and 7 were specifically deposited onto the cellulose surface through their family I cellulose-binding modules (CBM). Naturally glycosylated fungal laccase was then deposited onto the preformed glycoprotein layer pretreated by ConA, through the interaction of mannosyl moieties of fungal glycoproteins with the multivalent lectin. The formation of a cellulase-ConA-laccase composite was proven by direct and indirect determination of activity of immobilized laccase. In the absence of cellulases and ConA, no laccase deposition onto the cellulose surface was observed. Finally, basidiomycetous cellobiose dehydrogenase (CDH) was deposited onto the cellulose surface through the specific interaction of its FAD domain with cellulose. The obtained paste was applied onto the surface of a Clark-type oxygen electrode and covered with a dialysis membrane. In the presence of traces of catechol or dopamine as mediators, the obtained immobilized multienzyme composite was capable of the coupled oxidation of cellulose by dissolved oxygen, thus providing the basis for a sensitive assay of the mediator. Swollen amorphous cellulose plays three different roles in the obtained biosensor as: (i) a gelforming matrix that captures the analyte and its oxidized intermediate, (ii) a specific carrier for protein self-assembly, and (iii) a source of excess substrate for a pseudo-reagent-less assay with signal amplification. The detection limit of such a tri-enzyme biosensor is 50-100 nM dopamine.     

小麦种子中几丁质酶基因的扩增及序列分析

根据http://www.tigr.org中与小麦几丁质酶基因相关的序列TC187877,设计引物,分别从小麦品种Gamenya和苏麦3号中扩增到大小约为1 000bp的片段。经序列测定和软件分析比较,发现这些片段所编码的蛋白质氨基酸序列,都有CHITINASE-19.1和CHITINASE-19.2的基序,为第II类几丁质酶基因。扩增的核酸序列在GenBank上发表,登录号分别为AY973229和AY973230。     

青霉素酶发酵液的预处理和酶学特性

研究了在发酵液中添加絮凝剂对发酵液进行预处理,对蜡状芽孢杆菌（Bacillus cereus）产生青霉素酶的影响与酶学特性。实验结果表明,发酵液膜处理的难易程度与芽孢释放程度成正相关,在发酵液中添加0．02g／mL自制絮凝剂进行预处理,过滤效率提高10倍,而酶活损失仅5％;酶学特性研究结果表明,该酶最佳反应温度55℃时,酶的最适反应pH值7．0,金属离子锌、锰、镁对酶有激活作用,其浓度为0．25moL／L;酶热稳定性研究结果表明,在75℃下保温30min时,酶活损失85％。该酶在过量青霉素底物下,酶促反应为0级反应。     

3种荒漠盐生植物根系及根毛形态特征的比较研究

在水培条件下,针对梭梭、囊果碱蓬和钠猪毛菜3种荒漠盐生植物,研究它们苗期在不同盐浓度条件下根系和根毛形态的差异。结果表明：一定浓度的盐分可以促进3种盐生植物生长,但较高浓度的盐抑制其生长,特别是对根系生长的抑制作用更大。在同样盐浓度下,钠猪毛菜的生长最快,生物量也最大。在盐分浓度较低时,3种盐生植物的主根长和总根长都有所增加,与对照相比,囊果碱蓬增加的幅度较大,但高浓度的盐会抑制根系总长度的增加,其中囊果碱蓬较梭梭和钠猪毛菜抑制的程度轻。盐分对3种植物的根系平均直径没有显著的影响,但有减小的趋势。在水培条件下,梭梭和囊果碱蓬的根系上、中、下部分布的较均匀,而钠猪毛菜的根系中部比上部和下部有显著的增加,盐分对每种植物的根系的分布没有显著的影响。3种植物的根毛与根系生长环境中的盐浓度没有明显相关性;3种植物之间,根毛的长度和密度有显著差异。从根系和根毛的形态特征可以推断：囊果碱蓬比梭梭和钠猪毛菜具有较强的抗旱性和耐极薄能力;梭梭的耐盐能力较其它2种植物差,囊果碱蓬的耐盐性最强。