单核细胞增多性李斯特菌氨基肽酶Lmo1711体外表达及其酶活分析

对单核细胞增多性李斯特菌(简称单增李斯特菌)lmo1711基因编码的氨基肽酶进行克隆表达与纯化,并研究该重组蛋白的体外酶学特性。首先通过生物信息学分析预测Lmo1711与氨基肽酶家族成员的亲缘关系及关键活性位点的保守性。利用SWISS-MODEL模拟预测该蛋白的空间结构;构建Lmo1711原核表达载体并转化入E.coli Rosetta中,诱导表达重组目的蛋白,并利用镍离子亲和层析方法纯化目的蛋白;以氨基酸-对硝基苯胺偶联物为底物,Lmo1711通过水解底物N端氨基酸残基产生游离对硝基苯胺单体,405 nm处检测吸光值对该产物进行检测从而分析Lmo1711的酶学特性。在此基础上系统研究Lmo1711对不同氨基酸残基底物的催化特异性,及不同金属离子对该酶活性的影响。经原核表达纯化获得49.3 kDa的重组Lmo1711蛋白,与预测分子量一致;生物信息学分析推测Lmo1711属于M29氨基肽酶家族,且存在保守关键氨基酸活性位点(Glu250、Glu316、His345、Tyr352、His378、Asp380);酶活分析显示,Lmo1711具有较强的氨基肽酶活性,针对不同底物的结合和催化能力差异较大,对亮氨酸残基的亲和程度最高;Lmo1711氨基肽酶活性具有金属离子依赖性,Co~(2+)、Cd~(2+)、Zn~(2+)等多种金属离子均能显著增强其活性,其中Co~(2+)的激活效应最显著。本试验首次发现并证实,单增李斯特菌Lmo1711属于M29氨基肽酶家族成员,具有较强的催化活性,且对金属离子具有不同程度的依赖性。     

Catalase, but not MnSOD, inhibits glucose deprivation-activated ASK1-MEK-MAPK signal transduction pathway and prevents relocalization of Daxx: hydrogen peroxide as a major second messenger of metabolic oxidative stress

Overexpression of catalase, but not manganese superoxide dismutase (MnSOD), inhibited glucose deprivation-induced cytotoxicity and c-Jun N-terminal kinase 1 (JNK1) activation in human prostate adenocarcinoma DU-145 cells. Suppression of JNK1 activation by catalase overexpression resulted from inhibition of apoptosis signal-regulating kinase 1 (ASK1) activation by preventing dissociation of thioredoxin (TRX) from ASK1. Overexpression of catalase also inhibited relocalization of Daxx from the nucleus to the cytoplasm as well as association of Daxx with ASK1 during glucose deprivation. Taken together, hydrogen peroxide (H(2)O(2)) rather than superoxide anion (O(2) (*-)) acts as a second messenger of metabolic oxidative stress to activate the ASK1-MAPK/extracellular signal-regulated kinase (ERK) kinase (MEK)-mitogen-activated protein kinase (MAPK) signal transduction pathway.     

Rht10基因对‘鲁麦15’农艺性状和光合生理特性的影响

利用鲁麦15、Rht10鲁麦15、Ms2鲁麦15及Ms2+Rht10鲁麦15近等基因系为试验材料,研究了Rht10基因对小麦农艺性状、净光合速率(Pn)、蒸腾速率(Tr)等光合生理特性的影响.结果表明,Rht10使小麦的挑旗、抽穗、开花期均推迟了4～5 d;Rht10对鲁麦15和Ms2鲁麦15的降秆强度分别为53.77%和53.00%,穗长显著缩短(P<0.05),千粒重显著减少,但对有效穂数无显著影响(P>0.05).含有Rht10基因的材料与不含此基因的材料之间的光合生理参数普遍存在显著差异,但在不同时期不同参数差异正负不同;在灌浆后期,Rht10对Pn有显著正效应(P<0.05);从开花期到灌浆后期,Pn与气孔导度(Gs)、胞间二氧化碳浓度(Ci)总体变化趋势相同,且相关性显著,表明Pn在一定程度上受气孔因素的限制;在灌浆中后期,Rht10对Gs有明显的正效应;在抽穗和开花期,Rht10对Tr有明显的正效应;Rht10对叶片水分利用效率(WUE)在4个生育期有明显的负效应.由此可见,Rht10基因对小麦的生育期、株高、穗长都有明显的不利影响,对各项光合生理参数也有普遍正向或负向的影响,在利用Rht10进行杂交育种时应充分考虑此基因带来的不利效应.     

Trigoxyphins H and I: two new daphnane diterpenoids from Trigonostemon xyphophylloides

Two new daphnane diterpenoids (1 and 2), together with four known analogues (3-6) were isolated from Trigonostemon xyphophylloides. Their structures were elucidated by spectroscopic analysis. Compounds 1 and 2 were evaluated for in vitro cytotoxic activities against the SPCA-1 (human lung cancer) and BEL-7402 (human hepatocellular carcinoma) cancer cell lines. Trigoxyphin I (2) showed modest cytotoxicity against two tumor cell lines.     

Simulation of sequential batch reactor (SBR) operation for simultaneous removal of nitrogen and phosphorus

Modeling of the operation of sequential batch reactor (SBR) was performed to find out optimum design parameters for simultaneous removal of nitrogen and phosphorus in a small-scale wastewater treatment plant. The models were set up with material balances on SBR operation and Monod kinetics. The model parameters were obtained to best fit the experimental results in a small scale SBR. The models were useful in optimizing hydraulic retention time (HRT) and successfully simulated operations of SBR in a larger scale. Especially the model predicted well the reactions occurring in the filling period as well as the effect of dilution, and evaluated the performance of SBR process under diverse operating conditions.     

Influence of magnesium ions on biofilm formation by Pseudomonas fluorescens

Mg²⁺ can potentially influence bacterial adhesion directly through effects on electrostatic interactions and indirectly by affecting physiology-dependent attachment processes. However, the effects of Mg²⁺ on biofilm structure are largely unknown. In this study, Pseudomonas fluorescens was used to investigate the influence of Mg²⁺ concentration (0, 0.1 and 1.0 mM MgCl₂) on biofilm growth. Planktonic and attached cells were enumerated (based on DAPI staining) while biofilm structures were examined via confocal laser scanning microscopy and three-dimensional structures were reconstructed. Mg²⁺ concentration had no influence on growth of planktonic cells but, during biofilm formation, Mg²⁺ increased the abundance of attached cells. For attached cells, the influence of Mg²⁺ concentration changed over time, suggesting that the role of Mg²⁺ in bacterial attachment is complex and dynamic. Biofilm structures were heterogeneous and surface colonization and depth increased with increasing Mg²⁺ concentrations. Overall, for P. fluorescens, Mg²⁺ increased initial attachment and altered subsequent biofilm formation and structure.     

Clinical manifestation, imaging, and genotype analysis of two pedigrees with spinocerebellar ataxia

The objective of this study was to analyze the clinical manifestation, imaging characteristics, genotype, and the relationship between the three aforementioned parameters in two pedigrees suffering from spinocerebellar ataxia. To evaluate the clinical manifestation of the two pedigrees and to compare the characteristics, we performed the MRI analysis of some patients from both pedigrees, while 2 ml of the peripheral blood sample was collected for gene analysis. The gene analysis data showed that pedigree 1 was certified spinocerebellar ataxia type-2 (SCA2); the CAG repeats in the proband, proband’s mother, and proband’s brother were 44, 36, and 38, respectively. The MRI revealed brainstem cerebellar atrophy and “cross sign” and “ordinate sign” of pons. Pedigree 2 was certified SCA1; the CAG repeats of the proband, proband’s aunt, and proband’s asymptomatic cousin were 60, 51, and 52, respectively. The MRI revealed cerebellar atrophy in these individuals. We, therefore, concluded that it was difficult to diagnose the SCA subset solely through the clinical manifestation. The imaging characteristics analysis and final diagnosis depended basically on gene analysis data.     

重金属镉、锌在菹草叶细胞中的超微定位观察

水环境污染中十分突出的是重金属的污染 ,主要来源为流入物、渗漏和大气沉降 (Ｌａｒｃｈｅｒ ,1995 )。由于重金属污染物不但不能被微生物所分解 ,而且能在生物体内富集 ,并通过水生食物链的生物放大作用而对高营养级的生物甚至人类造成危害 ,因此日益引起人们的特别关注。许多研究从超微结构损伤和生理生化的角度研究了重金属对植物的毒害机制。施国新等 (2 0 0 0 )观察了重金属汞、镉污染对水生植物黑藻叶细胞的超微结构损伤。彭鸣等 (1991)研究了重金属镉、铅诱导的玉米超微结构的变化。李荣春 (2 0 0 0 )研究了Ｃｄ、Ｐｂ及其复合污染…