禽腺联病毒全基因组的克隆及感染性病毒的拯救

为了克隆禽腺联病毒(Avian adeno-associated virus,AAAV)全基因组用于构建基因转移载体研究,以鸡胚致死孤儿病毒(CELO)作为辅助病毒与AAAV共接种SPF鸡胚进行AAAV的增殖,将AAAV约4.7kb双链基因组DNA与pCR2.1载体连接,构建了含AAAV全基因组的重组质粒pAAAV并进行了测序。序列分析表明,AAAV YZ-1株的基因组为4684bp,两端具有141bp的末端倒置重复序列和Rep蛋白结合位点特征序列,与GenBank中收录的AAAV DA-1株和VR-865株的核苷酸序列同源性分别为95.0%和92.2%。将pAAAV质粒转染CELO病毒感染的鸡胚肝细胞系,获得了感染性AAAV病毒粒子,结果证明克隆的AAAV基因组中存在与病毒复制和包装相关的正确关键序列,可用于重组AAAV载体的构建。     

大熊猫犬冠状病毒部分S基因的克隆与全S基因序列分析

从重庆某动物园死亡的大熊猫肝脏中分离到一株病毒，经鉴定为犬冠状病毒（命名为CCV DXMV）。本实验根据CCV K378和CCV Insavc-1株基因序列设计合成了普通PCR引物SF1-SR1和SF2-SR2，以及半套式引物SF3-SR3、SR13、SF13、SF4-SR4和SF14，分段扩增了CCVDXMV株部分S基因，得到了368bp、792bp、737bp、1178bp和985bp5个基因片段。将纯化的RCR产物分别克隆到pGEM—T载体中，通过筛选和鉴定，获得了5个阳性重组质粒。将重组质粒送生物公司测序，将测得的基因序列与先前测定的该株病毒部分S基因序列，拼接成全S基因序列，并与其它株CCV及TGEV HN2002和FIPV79—1146株的S基因序列进行分析比较，绘制进化树。结果表明，该株病毒与CCV K378株的同源性最高，达到99．4％；而与CCV 23／03株的同源性最低，为56．9％；与FIPV和TGEV分别有90．4％和82．1％的同源性。     

茶花鸡种群遗传结构分析及其保种措施的探讨

利用14个微卫星标记对云南茶花鸡12个家系的等位基因频率、平均基因杂合度、平均多态信息含量以及家系间的亲缘关系进行了分析。结果表明:共检测到99个等位基因,每个位点的等位基因分布并不均匀,再对12个家系进行分析,将其聚为3类,一类为Z-1,Z-2,Z-3,Z-4,Z-5,Z-6,B-1,一类为Z-7,Z-8,Z-9,Z-10,矮脚系B-2自成一类,这表明了家系间的遗传相似性,同时我们对特殊的矮脚系B-2进行了相关的探讨,最后我们提出了对茶花鸡保种的相关建议。     

猪瘟病毒的分离鉴定及其E2基因序列分析

2004~2005年上海和江苏3家猪场发生仔猪急性死亡,用ELISA鉴别试剂盒证明为猪瘟感染。用PK15细胞分离病毒,传代至第3代无细胞病变,RT-PCR扩增E2基因为阳性,扩增片段经纯化后克隆入T载体并进行序列测定。通过DNAStar软件对3株病毒的基因序列进行了比较,同时构建了系统进化树,结果表明,三株病毒分离株均扩增出269bp片段并且核苷酸序列100%同源,这3株与Alfort、Shimen株、兔化弱毒株(HCLV)和Brescia4个标准毒株核苷酸序列同源性均在90%以上,说明在江苏、上海地区分离得到的3株猪瘟病毒与流行毒株在基因序列上没有发生大的变异。     

A comparison between multivariate Slash, Student's t and probit threshold models for analysis of clinical mastitis in first lactation cows

Robust threshold models with multivariate Student's t or multivariate Slash link functions were employed to infer genetic parameters of clinical mastitis at different stages of lactation, with each cow defining a cluster of records. The robust fits were compared with that from a multivariate probit model via a pseudo‐Bayes factor and an analysis of residuals. Clinical mastitis records on 36 178 first‐lactation Norwegian Red cows from 5286 herds, daughters of 245 sires, were analysed. The opportunity for infection interval, going from 30 days pre‐calving to 300 days postpartum, was divided into four periods: (i) ?30 to 0 days pre‐calving; (ii) 1–30 days; (iii) 31–120 days; and (iv) 121–300 days of lactation. Within each period, absence or presence of clinical mastitis was scored as 0 or 1 respectively. Markov chain Monte Carlo methods were used to draw samples from posterior distributions of interest. Pseudo‐Bayes factors strongly favoured the multivariate Slash and Student's t models over the probit model. The posterior mean of the degrees of freedom parameter for the Slash model was 2.2, indicating heavy tails of the liability distribution. The posterior mean of the degrees of freedom for the Student's t model was 8.5, also pointing away from a normal liability for clinical mastitis. A residual was the observed phenotype (0 or 1) minus the posterior mean of the probability of mastitis. The Slash and Student's t models tended to have smaller residuals than the probit model in cows that contracted mastitis. Heritability of liability to clinical mastitis was 0.13–0.14 before calving, and ranged from 0.05 to 0.08 after calving in the robust models. Genetic correlations were between 0.50 and 0.73, suggesting that clinical mastitis resistance is not the same trait across periods, corroborating earlier findings with probit models.     

Influence of ovariectomy and estrogen replacement on gene expression profile of myocardium in rats

AIM:To investigate the influence of ovariectomy and estrogen replacement treatment on profile of gene expression in myocardium by cDNA microarray,and to characterize the targeting genes of estrogen.METHODS:cDNA microarray containing 1 400 rat cDNAs was used to study the genes differentially expressed in myocardium between sham (Ⅰ),ovariectomy (Ⅱ,OVX) and estrogen replacement treatment (Ⅲ,OVX+E2) group.Then down-regulated genes in myocardium of OVX rats were further confirmed by RT-PCR.RESULTS:177 genes were differentially expressed in myocardium between sham and OVX rats,with 91 genes up-regulated and 86 genes down-regulated in OVX rats.164 genes were differentially expressed in myocardium between OVX and OVX+E2 rats,with 113 genes up-regulated and 54 genes down-regulated in OVX rats.There were 54 genes differentially expressed in OVX compared to sham and OVX+E2.They are involved in membrane channels and transporters (18),cell receptors (9),intracellular transducers/effectors/modulator (7) and metabolism (6).Most of the genes (45) were down-regulated in OVX rats and up-regulated in OVX+E2 rats.RT-PCR test confirmed the results of cDNA microarray.CONCLUSIONS:Long-term estrogen replacement may influence the expression of genes involved in membrane channels and transporters,cell receptors,intracellular transducers/effectors/ modulator and metabolism.Long-term estrogen replacement has some beneficial effects on ionic concentration and cardiac function which partially comes from the results of influence of expression on Na+,K+-ATPase and Na+/H+ exchanger.Estrogen has an inhibitory effect on the expression of dopamine receptor,which partially clarify the myocardial protection of estrogen.     

苯醚甲环唑·丙环唑乳油气相色谱分析

本文介绍了采用气相色谱法用HP-5毛细管柱,以邻苯二甲酸二正辛酯为内标物,氢火焰离子化检测器对苯醚甲环唑.丙环唑乳油进行定量分析。结果表明,苯醚甲环唑、丙环唑的线性相关系数为1.000 0、0.999 9;平均回收率为99.87%、99.98%;标准偏差为0.12、0.04;变异系数为0.82%、0.28%。     

1%蛇床子素水乳剂的高效液相色谱分析

采用反相高效液相色谱法对蛇床子素进行分离定量,本方法的标准偏差为0．004, 变异系数0．41％;平均回收率为99．86％;线性相关系数为0．999 99。     

10%吡嘧磺隆可湿性粉剂的液相色谱分析

本文所述应用液相色谱法测定吡嘧磺隆可湿性粉剂有效成分含量的测定方法,此方法采用甲醇、水和冰乙酸为流动相,使用C18柱、紫外检测器的高效液相色谱,经验证其添加回收率为100.09%,标准偏差为0.133,变异系数为1.33%,线性方程相关系数为0.9997。本试验方法操作简便,分离效果好,响应时间短,是理想的分析方法。