电针诱导心肌缺血大鼠延髓头端腹外侧区nNOS和iNOS差异表达

许多研究表明,延髓头端腹外侧区(rostral ventrolateml medulla,RVLM)的NO/NOS系统参与心血管活动的中枢调节.本实验以结扎Wistar大鼠左冠状动脉前降支法建立急性心肌缺血(acute myocardial ischemia,AMI)动物模型,观察针刺"内关"穴改善AMI大鼠的心功能作用,同时检测大鼠RVLM区神经元型一氧化氮合酶(neuronal nitric oxide synthase,nNOS)和诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)表达的变化,进而探讨针刺治疗AMI的中枢机制.实验观察显示,AMI大鼠心功能各项指标减弱,伴随外周血去甲肾上腺素(norepinephrine,NE)和脑钠肽(brain natriuretic peptide,BNP)水平显著升高,同时RVLM区nNOS阳性神经元数和nNOS mRNA表达升高,而iNOS水平则降低.针刺"内关"穴(Pe 6)(每天30 min,连续5天)改善心功能,降低AMI大鼠血清中NE和BNP的水平,同时升高iNOS并降低nNOS在RVLM的表达.以上结果提示,针刺治疗心肌缺血的同时可以调节iNOS/NO和nNOS/NO在RVLM的变化,这可能与针刺通过调节RVLM区的NO含量进而降低交感传出,从而改善AMI大鼠的心功能有关.     

An intramolecular disulfide bond is required for the thermostability of methyl parathion hydrolase, OPHC2

OPHC2, a methyl parathion hydrolase (MPH) from Pseudomonas pseudoalcaligenes C2-1 (CGMCC 1150), can degrade a wide range of organophosphate pesticides. Compared with other MPHs, OPHC2 exhibits high thermostability. Its thermostability mechanism, however, remains unknown. In the present study, sequence analysis demonstrated that two cysteines (Cys110 and Cys146) exist in OPHC2, but not in other MPHs. The three-dimensional structural model of OPHC2 performed by computer-assisted homology modelling revealed a potential stacking network with residues Cys110 and Cys146, which probably formed an intramolecular disulfide bond. Furthermore, both sodium dodecyl sulphate-polyacrylamide gel electrophoresis and thiol-titration analyses indicated that OPHC2 contains a disulfide bond. Substitution of the disulfide bond-forming cysteines with alanine, leucine or methionine residues substantially decreased the thermostability of OPHC2, suggesting that disulfide bond formation affects conformational stability. These results, combined with three-dimensional structural modelling, demonstrated that the formation of a C110-C146 disulfide bond may stabilise the conformation of OPHC2, contributing to its thermostability.     

柯萨奇B组病毒感染与胰岛素依赖型糖尿病关系的研究

探讨柯萨奇B组病毒(CBV)感染与胰岛素依赖型糖尿病之间的关系。用PT-PCR方法检测38例IDDM患者外周血中CBV-RNA,同时用ELISA法测定血清中CBV特异性IgM。结果显示,病例组38例患者外周血CBV-RNA阳性率为42.10％,血清中CBV-IgM阳性率为36.84％。对照组的阳性率分别为7.5％和5％。两组比较有显著性差别(P<0.05)。说明胰岛素依赖型糖尿病与柯萨奇B组病毒感染有关。     

PACAP protects neuronal differentiated PC12 cells against the neurotoxicity induced by a mitochondrial complex I inhibitor, rotenone

In vivo and in vitro studies have suggested a neuroprotective role for Pituitary adenylate cyclase activating polypeptide (PACAP) against neuronal insults. Here, we showed that PACAP27 protects against neurotoxicity induced by rotenone, a mitochondrial complex I inhibitor that has been implicated in the pathogenesis of Parkinson's disease (PD). The neuroprotective effect of PACAP27 was dose-dependent and blocked by its specific receptor antagonist, PACAP6-27. The effects of PACAP27 on rotenone-induced cell death were mimicked by dibutyryl-cAMP (db-cAMP), forskolin and prevented by the PKA inhibitor H89, the ERK inhibitor PD98059 and the p38 inhibitor SB203580. PACAP27 administration blocked rotenone-induced increases in the level of caspase-3-like activity, whereas could not restore mitochondrial activity damaged by rotenone. Thus, our results demonstrate that PACAP27 has a neuroprotective role against rotenone-induced neurotoxicity in neuronal differentiated PC12 cells and the neuroprotective effects of PACAP are associated with activation of MAP kinase pathways by PKA and with inhibition of caspase-3 activity; the signaling mechanism appears to be mediated through mitochondrial-independent pathways.     

聚乙烯醇生物降解研究进展

聚乙烯醇(PVA)是一种在纺织和化工行业中广泛使用的难降解的高分子聚合物。随着人们对纺织工业清洁生产的关注,如何在退浆工艺中就实现对PVA的生物降解、减少PVA废水的排放,并避免化学退浆过程中高温和氧化造成的棉纤维损伤,是近年来纺织生物技术领域的研究热点。由于PVA降解菌种类不多、培养周期长,PVA降解酶酶活不高、提取不容易等原因,使PVA的生化降解研究还局限在PVA降解菌的筛选、PVA降解酶的酶学性质研究等方面,PVA降解酶还未在纺织工业上得到应用。本文综述了近年来国内外在PVA降解菌筛选、PVA降解酶提取及酶学性质、PVA生化降解机理等方面的研究进展,并讨论了PVA生化降解研究中存在的问题及发展方向。     

湖南产五步蛇蛇毒磷脂酶A_2的热稳定性

本文测定了湖南产五步蛇蛇毒磷脂酶Ａ２在不同温度保温后的酶活性,结果表明此酶具有一定的热稳定性。     

一株抗锰土壤芽胞杆菌的分离鉴定与生物学活性分析

从山东崅屿采集的黄棕壤中分离得到一株具有抗Mn(Ⅱ)和Mn(Ⅱ)氧化双重活性的芽胞杆菌,其最高Mn(Ⅱ)耐受浓度达到130mmol/L,对Mn(Ⅱ)的氧化活性为3.3μmol/(L·d)。通过个体形态与培养特征观测、生理生化反应、G+Cmol%测定和16SrDNA序列比对分析等鉴定,确定该菌株为巨大芽胞杆菌(Bacillus megaterium),命名为MB283。该菌株在添加Mn(Ⅱ)(10mmol/L)条件下比不添加Mn(Ⅱ)表现出相对较快的生长速率。采用高温培养并结合0.01%SDS处理,从MB283菌株筛选到一株发生内生质粒消除的突变株MB287,具有与野生菌株类似的锰耐受活性,且对Mn(Ⅱ)的氧化活性与野生菌株相比无明显改变,表明野生菌株MB283中与锰抗性和锰氧化相关的基因可能是定位于该菌的染色体上。     

苏云金芽孢杆菌和大肠杆菌穿梭质粒的构建和特性研究

将苏云金芽孢杆菌中的pHTA1030质粒与大肠杆菌中的pJH101质粒重组后,构建出pBHGA重组质粒。此质粒通过逐步酶切,缺失后重组得到了14个分子量大小不同的衍生重组质粒。经过对pBHG1重组穿梭质粒在E.coli HB101和B.subttlis 168受体中表达的分析,证明了它带有B.thuringiensis(简写作B.t.)质粒的启动区、启始复制区和对热分离稳定区基因片段,并能高频转化B.t.受体细胞和高表达外源cat基因,同时具有对热分离稳定的特性。为B.t.基因工程体系提供了高效转化表达载体。     

多基因SNP位点及多种危险因素与老年汉族冠心病的关联分析

目的:探索老年冠心病多基因遗传易感性基础及相关的危险因素。方法:采用病例-对照研究方法,共入选老年汉族冠心病患者246例,非冠心病患者185例,纳入性别、年龄、吸烟,饮酒,高血压史、糖尿病史、高脂血症史、同型半胱氨酸、氨基末端脑钠肽前体、超敏C反应蛋白、抗凝血酶III、胆固醇、甘油三酯、高密度脂蛋白、低密度脂蛋白共15种危险因素与冠心病的关联性进行logistic回归分析。同时使用美国Sequenom高通量基因多态性分型技术研究了10种基因11个单核苷酸基因多态性(SNP)位点与冠心病的关联性。结果:15种危险因素中,发现增龄、高血压、抗凝血酶III(ATIII)下降是冠心病主要的危险因素,P<0.05。11个SNPs中3个SNP,血小板糖蛋白GP1BA rs2243093(-5T/C),血管紧张素转化酶ACE rs4332(547C/T)与ATIII rs2227589(893C/T)与老年汉族患者冠心病相关联。rs2243093(-5T/C)突变基因型CC与TT+AT比较,P=0.029(OR=3.41,CI:1.19-9.75);rs4332(547C/T)杂合型TC与CC+TT相比,P=0.003(OR=0.56,CI:0.38-0.82);rs2227589(893C/T),CT+CT与野生基因型CC相比较,P=0.003(OR=1.79,CI:1.22-2.63)。结论:增龄、抗凝血酶III下降、高血压是影响老年冠心病的主要危险因素,血小板、抗凝血系统、肾素-血管紧张素系统三种机制参与了老年冠心病的发生与发展。