摄入不同能量的围产期乳牛肝低密度脂蛋白受体mRNA丰度的比较

将30头健康、经产、处于围产期的黑白花乳牛随机分为3组，每组10头。从产前28d开始，低能量组乳牛饲喂《中国奶牛饲养标准（2000）》减少20％日粮（能量摄入80％），对照组乳牛饲喂《标准》日粮（能量摄入100％），高能量组乳牛饲喂《标准》增加20％日粮（能量摄入120％），产后各组乳牛均饲喂标准日粮。至产后第56d结束试验；采用内对照RT-PCR方法检测摄入不同能量的围产期乳牛肝活体组织低密度脂蛋白受体（LDLR）mRNA丰度。结果，不同能量组乳牛肝LDLR mRNA丰度产前至产后均呈现先升高后降低的趋势。100％和120％能量组肝LDLR mRNA丰度在产后14d达最大值，且产后均高于产前（产后56d除外，P〈0．01或P〈0．05）；而80％能量组产后1d即达到最大值，产前14d至产后14d，LDLR mRNA相对表达量显著高于100％和120％能量组；产后28～56d，120％能量组显著高于80％和100％能量组（P〈0．01）。表明围产期乳牛能量摄入水平对肝LDLR mRNA丰度有显著影响。     

感染16型蓝舌病病毒后绵羊部分细胞因子消长特点研究

本试验旨在探明绵羊感染16型蓝舌病病毒（Bluetongue virus type 16,BTV16）后细胞因子IFN-γ、IL-2、IL-4和IL-10的消长特点。用实时荧光定量PCR检测方法对感染BTV16后3只绵羊上述4种因子mRNA进行检测,同时设立阴性对照绵羊,并以0 d mRNA为基准,计算mRNA的相对表达量,同时检测病毒抗体效价、测量绵羊体温。结果显示,接种BTV16的3只绵羊均不同程度产生抗体和体温症状,4种细胞因子的mRNA在接种病毒2~4 d内均出现显著上升,其中IFN-γ峰值在2.58~27.84倍之间,IL-2峰值在5.24~17.19倍之间,IL-4峰值在2.16~3.43倍之间,IL-10峰值在15.78~48.77倍之间,个体上升幅度存在显著差异,4种细胞因子均在高水平持续6 d左右后逐渐下降。对照绵羊上述参数在正常范围内波动。本研究阐明了接种BTV16后绵羊细胞因子IFN-γ、IL-2、IL-4、IL-10在转录水平上的消长特点,为进一步深入开展BTV感染特征、宿主机体免疫机制研究提供参考。     

Study on Expression Difference of IGFBP-5 Gene in Different Tissues of Kazakh and Yanqi Horses

The study aimed to explore the mRNA expression pattern of insulin-like growth factor binding protein-5 (IGFBP-5) gene in different tissues of Kazakh and Yanqi horses.The expression of IGFBP-5 gene in different tissues of heart,liver,spleen,lung,kidney,small intestine,large intestine,cecum,intercostal muscles,longissimus dorsi muscles,brachialis muscle and gluteus in two horses were detected by Real-time quantitative PCR and compared the mRNA expression in the same tissues of two breeds.The results showed that the expression of IGFBP-5 in longissimus dorsi muscle and brachialis muscle of two breeds were significantly higher than other tissues including heart,liver,spleen,lung,kidney,small intestine,large intestine and cecum (P<0.05),and was the lowest in large intestine.The expression of IGFBP-5 in kidney,small intestine,large intestine,cecum,longissimus dorsi muscle and brachialis muscles of Yanqi horse were higher than in the same part of Kazakh horse,and among those in longissimus dorsi muscle and large intestine of Yanqi horse were extremely significantly higher than in the same part of Kazakh horse (P<0.01),and in small intestine and cecum of Yanqi horse were significantly higher than in the same part of Kazakh horse (P<0.05).The test was for further researching the biological function of IGFBP-5 gene,and it could provide a theoretical basis for genetic improvement of production performance of horse in our country.     

不同浓度赖氨酸对猪IEC碱性氨基酸转运载体mRNA表达的影响

本研究通过建立猪肠粘膜上皮细胞（Intestinal epithelial cells, IEC）原代培养的方法,研究了赖氨酸对体外培养的猪小肠粘膜上皮细胞碱性氨基酸转运载体mRNA表达的影响。用机械刮取初生仔猪小肠粘膜加胶原酶Ⅱ消化的方法,成功地在体外培养了新生仔猪IEC;分别用浓度为0.5 mM、2 mM、8 mM赖氨酸处理细胞96 h,提取细胞总RNA,采用实时荧光定量PCR方法,检测了碱性氨基酸转运载体在不同处理细胞中的表达丰度。结果表明：高浓度的赖氨酸对CAT-1 mRNA的表达有上调作用,差异极显著(P<0.01),并且呈剂量依赖效应;不同浓度的赖氨酸对CAT-2 mRNA的表达影响不大,8 mM和2 mM赖氨酸组CAT-2 mRNA的表达丰度均有低于0.5 mM的趋势,但二者之间差异不显著(P>0.05);不同浓度的赖氨酸对y+LAT1 mRNA表达影响差异不显著(P>0.05);高浓度的赖氨酸可提高4F2hc mRNA的表达;0.5 mM和2 mM赖氨酸组4F2hc mRNA的表达丰度均低于8 mM,且差异显著(P<0.05);0.5 mM和2 mM赖氨酸组之间差异不显著(P>0.05)。     

中华鳖脾脏雄激素受体mRNA 的原位杂交检测

采用原位杂交技术（In situ hybridization,ISH）研究雄激素受体（Androgen receptor,AR）mRNA在中华鳖（Pelodiseus Sinensis）脾脏的存在及细胞定位。取5只健康成年中华鳖的脾脏,进行石蜡切片。用地高辛标记的寡核苷酸探针对组织切片进行原位杂交（In situ hybridization,ISH）,检测雄激素受体mRNA在脾脏免疫细胞内的表达定位。结果显示,大部分雄激素受体mRNA阳性细胞呈圆形或椭圆形,数量少胞体大且呈不规则形。阳性细胞在脾脏动脉周围淋巴鞘（Pefiatefial lymphatic sheath,PALS）和红髓（Red pulp,RP）内分布密集。椭球周围淋巴鞘（Periellipsoidal lymphatic sheath,PELS）内AR mRNA阳性细胞分布极少。杂交阳性信号物质在细胞质和细胞核均有分布。在标记为阳性的中华鳖脾脏中,推测圆形的阳性细胞可能为B淋巴细胞,而另一部分胞体大且形状不规则的阳性细胞则可能为巨噬细胞。脾脏内AR mRNA的存在进一步证实雄激素可能是调节脾脏等免疫器官功能的因素之一。[中国水产科学,2008,15（2）：337—341]     

Isolation, Identification of Differentially Expressed Sequence Tags in the Backfat Tissue from Meishan, large White and Meishan×Large White Cross Pigs

In order to detect the molecular mechanism of heterosis in pigs, the mRNA differential display technique was performed to investigate the differences of gene expression in the backfat tissue from Meishan,Large White and Meishan×Large White cross pigs.Nine 3'-end anchored primers in combination with ten 5'-end arbitrary primers were used to perform the differential display PCR and nearly 3 000 reproducible bands were examined .Fifteen expressed sepuence tags that were differentially expressed were isolated and then isdentifide through semi-quantitative RT-PCR.BLAST analysis revealed that the fifteen expressed sequence tags (ESTs) were ntt homologous to any of the known porcine genes or ESTs. These novel ESTs were then submitted to GenBank.     

小鼠单个冷冻胚胎银染mRNA差异显示方法的建立

为建立单个冷冻胚胎银染mRNA差异显示方法,用小鼠单个4C鲜胚和冻胚RT—PCR产物,验证了冻胚和鲜胚实验结果的一致性。并从小鼠4C和8C冻胚的银染聚丙烯酰胺凝胶中回收了8C期的特异条带,通过再扩增、同收、克隆及酶切鉴定,克隆到了小鼠8C期胚胎差异表达的小鼠RP23—20A6基因。结果表明,所研究建立的单个冻胚银染mRNA差异显示方法具有可行性和有效性,可以用于动物早期胚胎基因表达的研究。     

复方中药成分与IL-2的免疫协同作用及对鸡细胞因子表达的影响

以淫羊藿多糖加蜂胶黄酮（方1）、人参皂苷加黄芪多糖（方2）组成2个复方，各加IL-2配合新城疫苗免疫雏鸡，分别于免疫后第7、14、21和28d用微量法检测血清抗体效价的动态变化；同时将2个复方各分3种剂量加入到培养的鸡外周血T淋巴细胞中，分别培养7和24h后用半定量RT-PCR方法检测淋巴细胞IL-2和IFN-γ的mRNA表达的变化。结果表明，2个复方单用或与IL-2合用均能显著提高抗体效价，而复方与IL-2的免疫协同作用不明显；方1的3个剂量和方2的高剂量能显著促进IL-2的基因表达，而2个复方的3个剂量能显著促进IFN-γ的基因表达，解释了复方中药成分与IL-2免疫协同作用不明显的原因。