表达丙型肝炎病毒NS3抗原的重组腺病毒构建及体外表达

为探索研制丙型肝炎疫苗的新途径,以期获得防治丙型肝炎的重组腺病毒减毒活疫苗,我们构建了表达丙型肝炎病毒(Hepatitis C virus ,HCV)非结构蛋白3(non structural protein 3,NS3)抗原的重组腺病毒RAd NS3,并检测其在体外表达。应用PCR从真核表达质粒pRC/NS3 中扩增编码HCV NS3 蛋白(329-935aa)的基因片段,定向克隆到重组腺病毒AdEasy-1系统的穿梭质粒pAdTrack CMV上,采用细菌内同源重组"两步转化法"构建携带HCV NS3基因的重组腺病毒基因组质粒pAd HCV NS3,转染293 细胞,成功包装出重组腺病毒RAd NS3,利用它有效地感染人肝癌细胞株HepG2,经RT PCR及免疫印迹等不同方法检测表明,被感染细胞能表达HCVNS3蛋白,为后续进行重组腺病毒在动物体内诱导抗HCV免疫应答能力的研究奠定了基础。     

温度对人肺癌细胞A549蛋白质表达的影响

肺癌蛋白质组的研究,有助于阐明其发病机制并对肺癌的防治有益。本文从研究人肺癌细胞热休克蛋白的表达情况出发,通过比较37℃、42℃和45℃培养条件下的人肺癌细胞A549总蛋白质的双向电泳图谱,获得3个温度敏感的差异蛋白点,依次命名为P1、P2、P3。对差异蛋白进行MALDI－TOF－MS分析和采用SWISS－PROT数据库中的Peptident软件检索后,初步鉴定P1与2种醛酮还原酶家族成员相匹配,P2可能为一种新蛋白,P3为锌指蛋白11A。     

DGGE技术在动物胃肠道微生物研究中的应用

变性梯度凝胶电泳(DGGE)是通过聚丙烯酰胺凝胶中变性剂浓度梯度的不同,将序列不同的DNA分开。目前,该技术已广泛应用于瘤胃微生物、肠道微生物等多样性研究。综述了变性梯度凝胶电泳技术的基本原理、优缺点及其在动物营养中的应用。     

黄海特定断面夏秋季大型底栖动物生态学特征

根据2001年8月和2002年8、9、10月共4个航次,在黄海特定断面的10个站采集到的大型底栖动物样品,采用相对重要性指数、物种多样性指数、物种均匀度指数、物种丰度指数,Bray-Curtis相似性系数聚类分析和MDS标序等方法,分析了该区域大型底栖动物的生态学特性。调查共得到定量样品36个,经分析鉴定有大型底栖动物182种,其中多毛类环节动物54种,软体动物29种,甲壳动物66种,棘皮动物17种,其它类群生物16种。相对于邻近海区,物种丰富度指数较小,物种多样性指数相似,均匀度指数较高;各航次中4、5号站种数明显较少,而生物量和栖息密度没有表现出一定的规律性。通过该断面大型底栖动物的研究发现,可将研究区域的大型底栖动物群落分为沿岸广温性群落、温带性群落和暖水性群落,各群落结构保持相对稳定性;主要底栖动物种类组成在2001年和2002年出现较大变化,而在2002年的8、9、10月的种类组成保持相对稳定。     

两株地理隔离的桦褐孔菌深层发酵产生抗氧化次生代谢产物的代谢组学分析

采用基于核磁共振代谢组学的方法研究了两株地理隔离的桦褐孔菌中国牡丹江菌株(IOM)和芬兰万塔菌株(IOV)在深层培养过程中主要代谢产物组成的差异。IOM和IOV两菌株在培养过程中积累菌丝体生物量、菌丝体酚类(TMP)以及三萜化合物(TMT)的过程相似,但IOM菌株产生较高水平的TMP。代谢产物组成动力学比较结果表明,IOM在第3天含有较高水平的硬毛素类似物、桦褐孔菌二糖和TMT,但在第5天均有所下降。导致这些差异的代谢产物为fuscoparianolD,21,24-羊毛甾-7,9(11),23-三烯-3β,22,25-三醇,羊毛甾-8,23-二烯-3β,22,25-三醇,23,24-环戊羊毛甾-3β,21,25-三醇-8-烯,桦褐孔菌二糖,phelligridinsC,D及H,甲基地花菌素A和C,地花菌素B,davallialactone及methyl davallialactone。这些代谢产物在IOV的提取物中均参与清除自由基,而在IOM的提取物中主要是酚类化合物参与自由基的清除。两菌株的这些差异反应了环境条件对桦褐孔菌生长及代谢的影响,即不仅影响其次生代谢产物的合成,还影响代谢产物在防止细胞氧化损伤过程中所起的作用。     

Effect of recombinant bovine somatotropin (bST) on follicular population and on in vitro buffalo embryo production

The objective of this study was to evaluate the effect of bovine somatotropin (bST) on ovarian follicular population in buffalo heifers and its influence on oocyte quality, recovery rates and in vitro embryo production. We tested the hypothesis that bST treatment in buffalo females submitted to an ovum pick-up (OPU) program would improve the number of follicles recruited, oocyte quality and in vitro embryo production. A total of 10 heifers were assigned into two treatment groups: group bST (n=5; receiving 500 mg of bST in regular intervals) and control group (n=5; without additional treatment). Both groups were subjected to OPU sessions twice a week (every 3 or 4 days), for a total of 10 sessions per female, although due to procedural problems, only the first five OPU sessions produced embryos. The number of follicles and the diameters were recorded at all OPU sessions. The harvested oocytes were counted and classified according to their quality as either A, B, C, D or E, with A and B considered good quality. Cleavage and blastocyst production rates were evaluated 2 and 7 days after in vitro fertilization, respectively. The bST treatment increased the total number of antral follicles (>3mm in diameter; 12.2 compared with 8.7; p<0.05) and of small antral follicles (<5mm; 9.1 compared with 6.5; p<0.05) per OPU session. The bST also tended to increase the number of oocytes recovered per session (5.2 compared with 4.1; p=0.07), and enhanced the percentage of good quality oocytes (48.8% compared with 40.6%; p=0.07). bST showed no effect on cleavage and blastocyst production rates (p>0.05). The significant effects of performing repeated OPU sessions were decreasing the follicular population (p<0.001) as well as the number of follicles aspirated (p<0.001), and oocytes recovered (p<0.02). In conclusion, bST treatment improves the follicular population, demonstrating its possible application in buffalo donors submitted to OPU programs.     

新型可移动RND家族外排泵TMexCD-TOprJ的研究进展

抗生素被认为是现代医学的基石之一,但包括抗生素在内抗菌药物的滥用也加速了可抵抗多种抗菌药物“超级细菌”的出现。耐药基因是导致细菌产生耐药性的关键因素,可通过质粒、转座子(transposon, Tn)、插入序列(insertion sequence, IS)等可移动元件(mobile genetic elements, MGEs)进行水平转移,严重威胁公共卫生安全。近年来,面对碳青霉烯类药物和多黏菌素耐药性的暴发,替加环素被视为人类面临多重耐药细菌感染的最后一道防线。近期发现了一种主要存在于质粒上的新型可移动外排泵基因簇tmexCD-toprJ,可编码耐药结节细胞分化家族(resistance-nodulation-cell division, RND)外排泵,排出菌体内包括替加环素在内的多种抗生素,大幅提升了细菌的耐药性。tmexCD-toprJ基因簇可以随质粒等可移动元件进行水平转移,已经传播至人、动物和环境中,给公共卫生健康造成了严重威胁。然而,目前人们对于其具体结构和功能作用机制等研究仍不透彻。本文系统总结tmexCD-toprJ耐药基因的分布特征、传播机制及外排泵结构等研究现状,并基于同一健康(One Health)理念提出了阻遏其扩散的措施,为减缓tmexCD-toprJ传播提供科学依据。     

A New Species of the GenusThermophis （Serpentes：Colubridae） from Shangri-La,Northern Yunnan,China, with a Proposal for an Eclectic Rule for Species Delimitation

The hot-spring snakes,Thermophis, were previously known only from the Tibet Autonomous Region and Western Sichuan, China. During the past two years, three adult hot-spring snakes （2 females, 1 male） were sampled in Shangri-La, northern Yunnan, China, thus expanding their known distribution region towards the southeast. This site is the southeastern-most corner of the Tibetan Plateau and the southernmost tip of the Hengduan Mountains （Mts.）. Phylogenetic analysis of mitochondrial （mt） and nuclear （n） DNA segments suggested that the three specimens belong to the genus ofThermophis. Morphologically, the new species is more similar toT. zhaoermii. However, it is distinguished fromT.zhaoermiiin the number of maxillary teeth （15）, distance between the two eyes/head width, rostral width/height, mental width/height, in one character limited to female： head width/length, and in four characters restricted to male： occurrence of the reduction from 10 to 8 （8 to 6, 6 to 4） scales in each dorsal row on the tail. There are differences in morphology, genetics （mtDNA, nDNA）, and geography between the putative new species andT.zhaoermii,the new species meets our proposed eclectic and feasible ＂four-differences＂ rule.     

中国人α珠蛋白基因3’HVR多态分布的研究

本文以3’HVR为探针,对来自全国17省市的51名无血缘关系的健康人进行P vuⅡ限制性片段长度的多态分析,发现3’HVR在片段大小的变异范围及基因频率的分布均有中国人的特点。结果表明,纯合子占15.7%,杂合子占84 .3%,102个等位基因共有34个长度不同的等位片段,大小从2.0-7.0kb之间呈连续分布, 主要分布在2.0-2.5kb及4.0-6.0kb,频率分别为0.43和0.34。并与刘国仰、余裕炉等的资料进行综合比较,初步揭示了中国人群中3’HVR的多态特点。 Abstract: 51 unrelated Chinese individuals from 17 different provinces and cities were typed for PvuII restiction fragment length polymorphism with the probe 3’HVR.The frequencies of homozygotes and heterozygotes were estimated as 15.7% and 84.3%,34 distinguishable allele sizes showed a continue distribution from 2.0-7.0kb.The main alleles of 2.0-2.5kb and 4.0-6.0kb gave the gene frequencies of 0.43 and 0.34 respectively.The preliminary characteristics of 3’HVR locus in Chinese population were revealed.     

云南锄足蟾科Pelobatidae二新种描述

1982年在云南景东哀牢山进行两栖类群落调查时,发现有两新种,现将其形态特征描述于后。正模标本保存在昆明动物研究所,副模标本保存在昆明分院生态研究室。景东齿蟾Oreolalax jingdongensis Ma,Yang et Li,新种(图1) 模式标本 正模:No.8200588,,云南省景东县哀牢山,海拔2400米,1982年5月24日采;配模:No.8200622,,1982年5月30日采;副模:4,1982年5月7