利用反向遗传技术拯救H9N2重组病毒

为研制高效特异的H9N2亚型AI疫苗,选取我国具有代表性的毒株A/chicken/Shanghai/10/01(H9N2)(简称SH10),以12质粒系统为基础,利用反向遗传操作技术构建了1株表面基因由SH10提供、内部基因由12质粒系统提供的重组病毒SH/PR8,为新型疫苗的研制提供了新的毒株。     

Multiple amino acid substitutions are involved in the adaptation of H9N2 avian influenza virus to mice

To explore adaptation of avian influenza virus to mice we previously performed serial lung-to-lung passages of the influenza A/Chicken/Jiangsu/7/2002 (H9N2) strain, resulting in the isolation of a variant influenza strain lethal for mice. We now report that virulence correlates with improved growth characteristics on mammalian cells and extended tissue tropism in vivo. Sequencing of the complete genomes of the wild-type and mouse-adapted viruses revealed 25 amino acid substitutions. Some were found to reiterate known substitutions in human and swine H9N2 influenza isolates. Functions affected include nuclear localization signals and sites of protein and RNA interaction, while others are known determinants of pathogenicity and host specificity such as the viral polymerase PB2 E627K substitution. These observations suggest that enhanced growth characteristics and modified cell tropism may contribute to increased virulence in mice. We conclude that multiple amino acid substitutions are likely to be involved in the adaptation of H9N2 avian influenza virus to mice.     

血红素氧合酶-2基因真核表达载体的构建

为构建血红素氧合酶-2（HO-2）真核表达载体pEF1α-HO-2-AcGFP,并观察其在小鼠脑血管内皮细胞中的表达情况。试验利用PCR技术从C57BL/6小鼠海马组织中扩增出HO-2基因cDNA序列,用双酶切法将此序列克隆到真核表达载体pEF1α-IRES-AcGFP上,构建HO-2的真核表达载体pEF1α-HO-2-AcGFP,重组载体经EcoRI和BamHI双酶切和测序鉴定。电穿孔法转染小鼠脑血管内皮细胞,48h后,实时定量PCR、Western blot检测HO-2在小鼠脑血管内皮细胞中mRNA和蛋白质水平的表达情况。结果表明,pEF1α-HO-2-AcGFP载体构建正确;重组载体转染小鼠脑血管内皮细胞后HO-2在mRNA水平表达量较空载体转染极显著增高（P〈0.01）,在蛋白水平表达量较空载体转染显著增高（P〈0.05）。表明HO-2基因的真核表达载体pEF1α-HO-2-AcGFP构建成功,为进一步研究其机制和功能奠定了基础。     

Is the metabolic syndrome a useful clinical concept in dogs? A review of the evidence

The metabolic syndrome is a set of risk factors for the development of type 2 diabetes, atherosclerosis, coronary heart disease and stroke in human beings. The term has recently been applied to dogs that exhibit components of the human metabolic syndrome, specifically visceral obesity, hypercholesterolaemia, hypertriglyceridaemia, hypertension and fasting hyperglycaemia. Obese dogs, like obese humans, are known to develop resistance to the glucose-lowering effects of insulin, and develop increased circulating concentrations of triglycerides, cholesterol and blood pressure. Unlike humans, however, obese dogs do not develop fasting hyperglycaemia or atherogenic hyperlipidaemia. Importantly, there is no evidence that dogs develop type 2 diabetes. Atherosclerosis, coronary heart disease and stroke are rare and not known to be associated with obesity in dogs. On the basis of current knowledge, the use of the term ‘metabolic syndrome’ in dogs does not appear to have merit.     

盐胁迫对星星草幼苗保护酶系统的影响

在盐胁迫下,星星草幼苗过氧化氢酶活性随着胁迫时间的延长而增强,并与盐浓度呈显著正相关。超氧化物歧化酶活性随着胁迫浓度的增强而增强,当超过一定浓度(3.0%Na₂CO₃)时则趋减弱,尔后随着胁迫时间的延长而增强。过氧化物酶活性的变化规律与超氧化物歧化酶活性相似,但变化幅度较小。丙二醛含量随着胁迫时间的延长和胁迫强度的增强而增加。     

犬新孢子虫NcSRS2基因原核表达质粒的构建

根据已发表的犬新孢子虫NcSRS2基因序列,设计1对含有EcoRⅠ和NotⅠ酶切位点的引物。以提取犬新孢子虫虫体基因组DNA为模板,应用PCR扩增获得NcSRS2 ORF基因片段,将此基因片段克隆到pMD18-T Simple载体上,用EcoRⅠ和NotⅠ双酶切该片段,回收得到含有两个酶切位点黏端的Nc-SRS2 ORF基因,将此基因片段克隆至相同酶切回收后的PGEM-4T-2原核表达载体中,获得重组质粒pGEX-NcSRS2,经PCR鉴定,限制性内切酶分析和克隆片段序列测定比较,证实了重组质粒的正确性。     

内蒙古草原N2O和CH4排放通量及其季节变化特征研究

１９９８年５－１２月,作者在内蒙古半干旱原用静态箱法进行原位Ｎ２Ｏ和ＣＨ４排放通量观测,分析了Ｎ２Ｏ、ＣＨ４排放量及其季节变化和相关过程,定时估计草原Ｎ２Ｏ和ＣＨ４收支在人类活动下的变化。重点讨论了内蒙古古草原型群落羊草,大针茅和退化草原Ｎ２Ｏ和ＣＨ４的排放通量及其季节变化特征。     

IGF2基因表达调控及其遗传变异在动物生长发育中的研究进展

胰岛素样生长因子2（insulin-like growth factor 2,IGF2）作为胰岛素类激素家族中重要成员之一,广泛参与机体众多生理代谢过程,在癌症发生、神经调节、糖代谢疾病、骨质疏松、肌肉发育和脂肪沉积等方面具有重要的作用,其功能行使主要通过与受体IGF1R和IGF2R结合或与胰岛素样生长因子结合蛋白IGFBPs和IGF2BPs竞争性结合发挥功能。鉴于IGF2基因在肌肉发育和脂肪沉积中的重要作用,发掘IGF2基因相关分子标记并解析其内在调控机理,在畜牧生产中具有重要的意义。研究发现,众多IGF2基因遗传变异与动物的生长发育之间存在显著相关关系,其可能通过影响IGF2基因印记、甲基化状态、转录因子结合或miRNA靶向结合型转录后调控等方式发挥作用。因此,文章综述了IGF2基因表达调控模式,包括IGF2与其受体的调控关系,基因印记与miRNA参与的表观遗传调控,转录因子对其的调节作用,遗传变异等方面的内容,以期为IGF2基因在动物生长发育调控相关研究中提供相应的借鉴,为分子育种提供有效线索。     

南阳牛IGF2基因第2外显子多态性及其与生长发育相关性研究

以126头纯种南阳黄牛为研究材料，利用PCR-RFLPs技术对胰岛素样生长因子2（IGF2）第2外显子的多态性及其与生长发育的相关性进行了分析。结果表明：南阳牛在该位点的遗传多态性丰富。检测到2种等位基因A和B，频率分别为0．6865和0．3135，A等位基因在群体中是优势等位基因；BB型个体的6、12、18、24月龄体斜长和胸围，12、18和24月龄的坐骨端宽，初生、6月龄和24月龄体重等体征性状显著地优于AA型；6月龄体斜长、12月龄的胸围、18月龄和24月龄坐骨端宽也显著优于杂合型。初步推断JGF2基因是非常有价值的辅助选择标记，可为南阳牛肉用型方向选择提供理论依据。     

仔猪产气荚膜梭菌肠毒血症病原诊断及疫苗研究IV．肠毒血症C型疫苗生产用菌种的确定

试验证明用单一菌株C59-2生产C型产气荚膜梭菌肠毒血症干粉灭活疫苗,其效力可以达到《规程》要求,并且简化了生产工艺,降低了生产成本,提高了生产效率。疫苗免疫的兔血清可分别中和原来3个制苗用菌株所产生的外毒素。