肝素或钙离子载体诱导牛、羊冻精体外获能的研究

本文采用肝素或钙离子载体(I-A)诱导牛、羊冻精体外获能,根据对去透明带仓鼠卵的穿透情况评定获能效果,并用台盼蓝·姬姆萨(TG)染色观察了精子的死活及顶体状态.结果发现,牛、羊冻精经肝素或I-A处理后与卵子一起培养,2 h穿透仓鼠卵,4 h开始形成雄原核,6 h形成发育良好的雄原核.牛冻精用50μg/mL肝素及0.1μmol/L I-A处理效果最好,穿透率分别为72.8％和92.3％;羊则以20μg/mL肝素及0.3μmol/L I-A为宜,穿透率分别为73.1％和68.O％.咖啡因与I-A或肝素并用均有协同作用,能提高精子的穿透能力,并能促进卵内原核的发育.肝素和I-A均能在体外诱导牛、羊冻精的顶体反应,有顶体反应活精子百分率与穿透率呈强正相关,羊冻精比较脆弱,获能处理后活力较低.     

两个黄芪亚种染色体数目与核型分析

前人研究认为膜荚黄芪（Astragalus membranaceus（Fisch.）Bge.）与蒙古黄芪（Astragalus membranaceus（Fisch.）Bge.var.mongholicus （Bge.）Hsiao）染色体数目为2n=16。本试验采用根尖染色体常规制片方法,观察了膜荚黄芪与蒙古黄芪种子根尖有丝分裂相。结果显示：膜荚黄芪与蒙古黄芪染色体数目为2n=18,这2种植物各有4条小染色体。膜荚黄芪染色体核型公式为K（2n）=2x=18=4sm+14m,核型类型为2B型。蒙古黄芪染色体核型公式为K（2n）=2x=18=6sm+12m,核型类型为2C型。     

Acute phase proteins in cattle and swine: A review

The major acute phase proteins (APPs) in cattle are haptoglobin (Hp) and serum amyloid A (SAA), and in swine, are Hp, SAA, C-reactive protein (CRP), and Pig major acute phase protein (Pig-MAP). Many methodologic assays are presently available to measure these parameters, which are still being improved to increase their specificity, sensitivity, user-friendliness, and economic availability. In cattle, the main applications are the diagnosis and monitoring of frequent diseases such as mastitis and metritis in dairy cows and respiratory problems in young calves. In pigs, APPs are useful in the control of bacterial and viral infections, and they may be used at the slaughterhouse to monitor subclinical pathologies and improve food safety. The utility of APP in animal production must not be forgotten; optimization of protocols to improve performance, welfare, and nutrition may benefit from the use of APPs. Other sample types besides serum or plasma have potential uses; APP determination in milk is a powerful tool in the control of mastitis, saliva is a non-invasive sample type, and meat juice is easily obtained at the slaughterhouse. Increasing our knowledge of reference intervals and the influence of variables such as age, breed, sex, and the season is important. Finally, worldwide harmonization and standardization of analytical procedures will help to expand the use of APPs.     

猪链球菌2型分泌性核酸酶A(SsnA)活性检测及其基因序列的克隆与测序分析

对8株猪链球菌2型重庆分离株的核酸酶A全基因进行克隆测序,结果表明该基因长度为3 126 bp,与Gen-Bank发表的唯一的该基因的序列相比,核苷酸同源性高于98%,推导的氨基酸同源性高于94%。根据核酸酶A基因的测序结果建立扩增片段长度为301 bp的PCR检测方法,并用甲苯胺蓝DNA酶琼脂对猪链球菌分泌性核酸酶A的活性进行检测。检测了从病猪内脏分离的猪链球菌致病株35株,33株核酸酶A基因PCR检测阳性(阳性比例为94.3%),其中有24株分泌活性检测为阳性(阳性比例68.6%);正常猪扁桃体分离株14株,PCR检测为阳性的10株(阳性比例71.4%),其中有核酸酶A分泌活性为8株(阳性比例57.1%),检测菌株中有2型猪链球菌44株,38株扩增出核酸酶A基因的片段PCR检测阳性并且分泌活性为阳性的有32株,猪链球菌1型、7型、9型、13型、1/2型各1株,均能扩增出核酸酶A基因的片段,但只有13型猪链球菌有核酸酶A分泌活性。对猪链球菌在不同生长时期核酸酶活性的检测显示,猪链球菌在培养4、8、16、32、48 h均有核酸酶A的分泌,并且C a2 和M g2 为分泌性核酸酶A的活性必需因子。     

Genetics of equine bleeding disorders

Genetic bleeding disorders can have a profound impact on a horse's health and athletic career. As such, it is important to understand the mechanisms of these diseases and how they are diagnosed. These diseases include haemophilia A, von Willebrand disease, prekallikrein deficiency, Glanzmann's Thrombasthenia and Atypical Equine Thrombasthenia. Exercise-induced pulmonary haemorrhage also has a proposed genetic component. Genetic mutations have been identified for haemophilia A and Glanzmann's Thrombasthenia in the horse. Mutations are known for von Willebrand disease and prekallikrein deficiency in other species. In the absence of genetic tests, bleeding disorders are typically diagnosed by measuring platelet function, von Willebrand factor, and other coagulation protein levels and activities. For autosomal recessive diseases, genetic testing can prevent the breeding of two carriers.     

新型重配A（H7N9）流感病毒NA基因序列分析

试验旨在分析2013年新型重配A（H7N9）流感病毒分子流行病学特点。作者从GenBank数据库下载不同分离宿主的流感毒株NA全基因序列,应用分子生物学软件进行遗传进化分析。结果显示,2013年新型重配A（H7N9）流感毒株NA蛋白颈部氨基酸发生缺失,其唾液酸结合（HB）位点发生一定程度适应性变化,NA蛋白糖基化位点为7个,NA蛋白酶活性中心未发生变异。与A/Hangzhou/1/2013（H7N9）株中NA片段的核苷酸同源性较高的前10个序列均分离自亚洲5个国家的禽类,同源性达96%以上。至于此次新型重配A（H7N9）流感毒株NA基因是否是禽传给人尚不清楚,还有待进一步研究。     

白细胞介素-6对猪脂肪细胞脂肪分解的影响

为了研究白细胞介素(Interleukin,IL)-6对猪脂肪细胞脂肪分解的影响及其分子机制,分化的猪脂肪细胞用不同浓度的IL-6(0、25、50、75、100 ng·mL-1)处理24或48 h.通过测定培养液中的甘油浓度检测脂肪细胞的脂解率;采用形态学观察检测脂肪细胞中甘油三酯积聚量的变化;分别用RT-PCR和Western blot检测脂肪细胞中perilipin A、PPAR γ2(Peroxisome proliferato-activated receptor-gamma2)、HSL(Hormone-sensitive lipase)和AT-GL(Adipose triglyceride lipase)的mRNA及蛋白表达.结果显示,IL-6以剂量和时间依赖性的方式刺激猪脂肪细胞的脂肪分解,同时PPAR γ2和perilipin A的mRNA及蛋白表达被显著下调;HSL的mRNA表达达著上调,但其蛋白表达水平并未发生显著改变;ATGL的mRNA和蛋白表达均未发生显著改变.上述研究结果表明,IL-6通过抑制PPAR γ2及其靶基因perilipin A的表达直接刺激猪脂肪细胞的脂肪分解.     

A cytotoxic cardenolide and a saponin from the rhizomes of Tupistra chinensis

A new cardenolide tupichinolide (1) and a new steroidal saponin tupichinin A (2), together with seven known compounds, were isolated from the rhizomes of Tupistra chinensis. Their structures were established using spectroscopic analysis and chemical methods. Compound 1 was the first cardenolide isolated from Tupistra chinensis and exhibited potent cytotoxicity against five human cancer cell lines: HL-60, SMMC-7721, A-549, MCF-7 and SW480.     

阔叶猕猴桃遗传转化技术参数的优化

为了提高阔叶猕猴桃的遗传转化效率,以阔叶猕猴桃叶柄为试材,通过根癌农杆菌介导法进行了遗传转化技术参数的研究。结果表明,叶柄预培养3～4d、用农杆菌悬浮液(D600nm值0.5)感染10min、共培养48h、共培养时在培养基中加入200μmol/L乙酰丁香酮处理可以获得较高的gus基因表达率。在供试的1200个叶柄中获得了49个抗性芽,转化频率为4.1%。对转基因抗性材料进行PCR检测和gus基因组织化学染色,证实了外源基因已整合到阔叶猕猴桃的基因组中,并得到稳定表达。