雌性骆驼生殖组织β-防御素-1基因的克隆及其组织表达量的检测

从雌性骆驼输卵管、子宫、子宫颈、阴道组织中提取总RNA，根据已发表的骆驼β-防御素-1基因的cDNA序列设计合成引物，采用RT—PCR扩增出了骆驼β-防御素-1基因；将扩增产物克隆于pBlueselect T载体后进行了序列分析。以伊肌动蛋白（pactin）基因作为内参，对扩增的β-防御素-1基因进行琼脂糖凝胶电泳后，应用凝胶成像分析系统，推断出了不同组织中β-防御素-1基因的表达量。结果，从雌性骆驼生殖各组织上皮均获得了203bp的β-防御素-1基因的扩增片段，且β-防御素-1基因在雌性骆驼生殖道各组织内的表达量不同。结果表明，β-防御素-1在雌性骆驼生殖组织的先天免疫中起重要作用。     

蒙古绵羊β-防御素基因相对表达水平实时定量PCR方法的建立

为了利用SYBR Green Ⅰ实时荧光定量PCR技术对蒙古绵羊雌性生殖道β-防御素基因表达水平进行相对定量测定,建立蒙古绵羊β-防御素基因相对表达水平的实时荧光定量PCR方法,试验根据GenBank中羊β-防御素基因序列设计合成引物,进行实时荧光定量PCR,同时以β-Actin为内参基因对β-防御素基因进行均一化处理,利用荧光阈值(Ct值)计算β-防御素基因的相对表达量.结果表明:扩增产物为β-防御素;利用SYBR Green Ⅰ实时荧光定量PCR技术可以测定蒙古绵羊β-防御素基因表达水平相对含量.     

绵羊妊娠期生殖组织Flt-1基因的克隆及其组织表达量的检测

试验从雌性绵羊妊娠期不同阶段的输卵管、子宫内膜、卵泡组织中提取总RNA,根据已发表的绵羊Flt-1基因的cDNA序列设计引物,以β-Actin基因为内参,采用RT-PCR扩增绵羊Flt-1基因,将扩增产物克隆于载体后进行序列分析,并应用Kodak Science 1 D、Bandscan图像分析软件,推断出不同组织中Flt-1基因的表达量。结果表明,从雌性绵羊妊娠期不同阶段生殖道各组织上皮均获得82 bp的Flt-1基因的扩增片段,且Flt-1基因在雌性绵羊生殖道各组织内的表达量不同。说明Flt-1基因对绵羊的妊娠维持起着重要的作用。     

绵羊妊娠期生殖组织VEGF基因的克隆及其组织表达量的检测

从雌性绵羊妊娠期不同阶段的输卵管、子宫内膜、黄体组织中提取总RNA,根据已经发表的绵羊的Flt-Ⅰ基因的cDNA序列设计合成引物,采用RT-PCR扩增出绵羊VEGF基因;将扩增产物克隆于裁体后进行序列分析,以基因为内参,对扩增的VEGF基因进行琼脂糖凝胶电泳后,应用,推断出不同组织中Flt-Ⅰ基因的表达量.结果表明:从雌性绵羊妊娠期不同阶段生殖道各组织上皮均获得82 bp的VEGF基因的扩增片段,且VEGF基因在雌性绵羊生殖道各组织内的表达量不同.说明VEGF基因对绵羊的妊娠维持起着重要的作用.     

绵羊生殖道中ghrelin基因的克隆及其表达

从雌性绵羊输卵管、子宫体、子宫颈、阴道组织中提取总RNA，根据已获得的绵羊ghrelin基因cDNA序列设计特异性引物，采用RT—PCR方法扩增出了绵羊ghrelin基因；将扩增产物克隆于pMD19-T载体后进行测序。以β-肌动蛋白（β-actin）基因作为内参，采用半定量RT—PCR法扩增ghrelin基因，经琼脂糖凝胶电泳后，应用凝胶成像分析系统计算雌性绵羊不同生殖道组织中ghrelin基因的表达量。结果显示，从雌性绵羊生殖道各组织均扩增出了233bp的ghrelin基因片段；ghrelin基因在子宫体的表达量最高，输卵管内次之，子宫颈和阴道内最低。表明，ghrelin对雌性绵羊生殖系统的调节及生殖激素的分泌等具有重要作用。     

蒙古绵羊β-防御素-1全长cDNA序列的扩增与分析

本研究以蒙古绵羊瘤胃为材料,提取瘤胃上皮组织总RNA,通过逆转录-聚合酶链式反应(RT-PCR)扩增出蒙古绵羊β-防御素-1(mgSBD-1)基因的核心片段序列,再应用5ˊ和3ˊ快速扩增cDNA末端(RACE)方法扩增蒙古绵羊β-御素-1基因的cDNA末端序列,最终获得mgSBD-1基因的全长cDNA序列.结果显示,该全长cDNA序列为325个碱基[不含poly(A)],5ˊ非翻译区为31个碱基,编码区(192个碱基)可编码64个氨基酸,3ˊ非翻译区为99个碱基[不含poly(A)].     

蒙古绵羊β-防御素-1cDNA的扩增及其序列分析

为了获得蒙古绵羊β-防御素-1（sBD-1）的cDNA序列,从蒙古绵羊瘤胃的上皮组织中提取总RNA,采用RT-PCR技术扩增出sBD-1的cDNA。结果 sBD-1的cDNA长为300bp,该cDNA包含由192bp组成的开放读码框（ORF）,ORF编码64个氨基酸的前原防御素。经过DNA序列分析。证实所扩增的cDNA为sBD-1。     

绵羊防御素(sBD-1)cDNA的克隆及其植物中表达载体的构建

为了获得在植物中表达的绵羊防御素 ,从绵羊舌表皮组织中分离提取总RNA ,经逆转录PCR (RT PCR)扩增出绵羊防御素sBD 1全长cDNA ,回收扩增产物连接到克隆载体 ,测序分析表明该cDNA为 2 1 5bp ,与报道序列完全一致。将该序列替换插入质粒PBI1 2 1中的GUS编码序列 ,构建植物表达载体PBIC 35SsBD1 ,为进一步进行转基因植物和绵羊防御素功能的研究奠定了基础。     

鸡β-防御素Gal-1 cDNA的克隆及序列分析

利用细胞总RNA提取试剂盒,从1月龄SPF鸡白细胞中分离提取总RNA,经过反转录PCR(RT-PCR)扩增出鸡β-防御素(Gal-1)cDNA片段。PCR产物经凝胶回收纯化后与pMD18-T载体连接,转化感受态JM109细胞,在含X-gal、IPTG的LB平板上筛选阳性克隆,经菌落PCR与质粒限制性酶切鉴定后,对目的片段进行测序。结果表明RT-PCR扩增出的cDNA片段ORF大小为198bp,用Blast程序进行检索比较表明该序列与Genbank中发表的鸡Gal-1 cDNA编码区序列同源性高达99.5%。该cDNA序列编码65个氨基酸,包括信号肽、前片段和成熟肽,成熟肽由40个氨基酸残基组成。     

绵羊β-防御素在毕赤酵母中的表达及活性鉴定

根据GenBank中绵羊β-防御素(SBD-1)的全长cDNA序列设计合成1对引物,PCR扩增SBD-1基因并与T载体成功连接,然后将其成熟肽编码片段亚克隆到毕赤酵母表达载体Ppic9k上,构建真核表达质粒Ppic9k-mSBD-1,将其在毕赤酵母GS115中进行分泌表达,表达产物纯化后进行活性鉴定。结果表明,mSBD-1对大肠杆菌和金黄色葡萄球菌均有抑制作用,而且对金黄色葡萄球菌的抑制作用更为明显。     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

中华人民共和国农业部公告 第267号

为贯彻落实《兽药生产质量管理规范》(简称《兽药GMP》),进一步推动兽药GMP实施进程,我部制定了《兽药生产质量管理规范检查验收办法》,现予公告。本公告自2003年6月1日起施行。附件:兽药生产质量管理规范检查验收办法二○○三年四月十日第一章 总则 第一条 为推动《兽药生产质量管理规范》(以下简称兽药GMP)的实施,规范兽药GMP检查验收工作,制定本办法。 第二条 农业部负责全国兽药GMP管理和检查验收工作;负责制修订兽药GMP检查验收管理规定;负责兽药GMP检查员队伍建设和监督管理工作,负责国际兽药贸易中GMP互认工作。 …     

鸡败血支原体垂直传播模式及其阻断方法

以国际标准强毒Ｒ株人工感染非免疫产蛋鸡,定时扑杀,分别从鼻窦、眶下孔、气管、肺、气囊、卵巢和输卵管分离ＭＧ,并收集感染鸡所产蛋分离ＭＧ。结果表明,人工感染４８小时后上、下呼吸道及肺已被全面感染,９６小时气囊已被感染,１２０小时输卵管已能分离到ＭＧ,卵巢始终分离不到ＭＧ。人工感染鸡自１４４小时便能在其所产蛋中分离出ＭＧ。药物治疗能在７２小时内消除感染,油乳剂苗则需２４天后逐渐降低蛋内ＭＧ分离率,药物卵内注射、种蛋药浴、高温处理均能杀死卵内ＭＧ,但以研制的种蛋浸泡剂药浴效果为最好。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.