鸡Hsc70基因5′侧翼区SNP与耐热性状的相关分析

为研究Hsc70基因作为鸡抗病育种中一种分子标记的可能性,本试验利用克隆测序技术对清远麻鸡、广东温氏矮脚黄鸡、灵山鸡和隐性白羽洛克鸡4个品种共20个个体Hsc70基因的5′侧翼区进行多态性筛查,发现10处变异,包括8个SNPs和2个插入/缺失突变。通过分析,选取C.-521A〉C位点,利用PCR-RFLP方法分析了此位点在灵山鸡和隐性白羽洛克鸡群体中的基因型分布,所得分型结果与测定的耐热性状（T3、皮质酮、CD3＋、CD4＋和CD8＋）进行关联分析,结果表明：常温状态下（15℃）,在隐性白羽洛克鸡群体中,位点C.-521A〉C与CD3＋、CD4＋和CD8＋显著相关（P〈0.05）,其中AA基因型个体CD3＋值显著低于AC基因型个体,而CD4＋与CD8＋值显著高于AC基因型个体;热应激状态下（35℃）,在灵山鸡群体中,位点C.-521A〉C与CD8＋极显著相关（P〈0.01）,CC基因型个体CD8＋值极显著高于AA和AC基因型。     

奶牛热应激与HSP70的表达

HSP70是应激条件下所诱导出的最为显著的热休克蛋白之一,是应激综合反应的代表。作者简要地介绍了HSP70的分类、结构、生理功能,及在热应激条件下奶牛HSP70的转录、翻译及检测方法。     

四种牛分枝杆菌特异性蛋白融合表达及在牛结核病诊断中的临床应用

以原核表达的MPB70-MPB83-CFP10-ESAT-6融合蛋白作为诊断抗原,建立牛结核病抗体检测间接ELISA(iELISA)。该iELISA与导致常见牛病的非相关抗原无交叉反应。用iELISA检测90份健康牛血清,确定样本阴阳性临界值(S/P)为0.17。与商品化胶体金试剂条(ICG)平行检测150份临床奶牛血清样本,结果与ICG的符合率为93.33%(140/150)。以结核菌素皮内试验(TST)为参考方法,检测了华中地区四个奶牛场的560头中国荷斯坦奶牛和90份进口奶牛结核阴性血清,结果iELISA与TST的总符合率为87.32%(489/560)。对其中22头奶牛进行鼻拭子分菌检验,4头分菌阳性奶牛的血清抗体检测也为阳性,iELISA与细菌培养的总符合率为77.27%(17/22)。以TST为参考方法,iELISA的敏感性和特异性分别为72.37%和89.67%。     

cDNA of Canine Heat Shock Protein 70 (HSP70)

Full-length canine HSP70 cDNA was sequenced and the expression of HSP70 mRNA was investigated. The full-length cDNA sequence of the HSP70 gene (2322 bp) contained a single long open reading frame (1920 bp) coding a protein of 640 amino acids. The amino acid sequence of the canine HSP70 gene shared about 90-95% sequence similarity with bovine, human and mouse HSP70 proteins. Southern blot analysis with HSP70 probe gave three distinct bands of 9.4 kb, 5 kb and 4.4 kb in BamHI digests and two distinct bands of 19 kb and 4 kb in EcoRI digests. Canine HSP70 mRNA was detectable in canine peripheral blood mononuclear cells and stomach but not in liver, kidney, spleen, small intestine, large intestine and skin of dogs.     

应激宁对HSP70在大鼠应激性溃疡中表达的影响

为了探讨热休克蛋白70(HSP70)在应激性溃疡中表达的变化及应激宁对其变化的影响,选用Wistar大鼠水浸-束缚应激(WRS)4 h的方法,建立应激性溃疡模型.用免疫组织化学方法检测胃黏膜组织HSP70的表达.结果表明,HSP70的表达主要分布在胃腺区,对照组大鼠胃黏膜组织中有散在的表达,应激组中HSP70阳性细胞数目较应激宁组增多,两者具有明显的差异(P＜0.05),而且,应激组和应激宁组中HSP70阳性细胞数目均比对照组明显增多(P＜0.01).表明应激宁对WRS后胃黏膜组织HSP70的表达具有调节作用,这也表明应激宁具有抗应激的作用.     

Effects of oxypolygelatin and dextran 70 on hemostatic variables in dogs

Objective To evaluate and compare coagulation variables following the administration of oxypolygelatin and dextran 70 to clinically healthy dogs. Study design Randomized cross‐over experimental study. Animals A total of eight healthy adult female Beagles aged 2–4 years old and weighing 11.8 ± 2.7 kg. Methods The dogs received a 15‐minute intravenous (IV) infusion of 5 mL kg^?1 oxypolygelatin or 10 mL kg^?1 6% dextran 70. Before (PRE) and at 2, 5, and 24 hours after administration, packed cell volume (PCV), total solids concentration (TS), prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen concentration (FIB), platelet numbers (Plat), factor VIII coagulant activity (VIII:C), von Willebrand factor antigen concentration (vWf:Ag) and platelet function and buccal mucosal bleeding time (BMBT) were measured. Platelet function was assessed using aggregation and by measuring ATP release from aggregating platelets over 6 minutes, with 20, 10, and 5 µm ADP and 5 and 10 µg of collagen mL^?1 as platelet activation agonists. Results All baseline values were within our normal ranges, except for one dog that had low vWf:Ag PRE values prior to both dextran and oxypolygelatin administration. Following dextran and oxypolygelatin administration, the PCV and TP were significantly (p < 0.05) decreased. Plat, FIB, and vWf:Ag decreased, while BMBT and VIII:C increased following dextran administration. Dextran also caused a significant decrease in platelet aggregation in response to ADP. Oxypolygelatin caused a significant decrease in vWf:Ag, Plat, and FIB compared to PRE values. The total amount of ATP released, standardized to platelet number, did not vary significantly for either group at any sampling time from PRE values. No significant changes from PRE values were noted at any time in either group for PT or APTT. Conclusion At the doses administered, both dextran and oxypolygelatin can interfere with hemostatic variables in healthy dogs, but dextran's effect is more profound and prolonged when compared to oxypolygelatin. Clinical relevance Oxypolygelatin causes fewer hemostatic abnormalities when compared to dextran, making it a superior colloid for administration at the doses tested.     

本地毛形线虫HSP70基因的克隆与序列分析

根据GenBank中已发表的布氏旋毛虫HSP70基因序列设计了1对引物，用Trizol法从本地毛形线虫肌幼虫虫体中提取总RNA，用RT-PCR方法扩增了HSP70基因，将目的基因克隆至pMD18-T载体，转化大肠杆菌DH5α。重组质粒用PCR、限制性内切酶BarnHⅠ和HindⅢ进行单、双酶切鉴定。测序结果表明，成功克隆了本地毛形线虫HSP70基因。序列分析表明，HSP70基因比较保守，不同物种之间氨基酸的同源性在40％～80％。与布氏旋毛虫HSP70序列相比，CDS区多出9个碱基，但核苷酸序列和氨基酸同源性分别为98%和94％，存在1个糖基化位点和1个潜在的信号肽位点。