牛传染性鼻气管炎病毒IBRV-LN01/08株的分离鉴定

从辽宁省某牛场出现疑似牛传染性鼻气管炎症状的牛鼻拭子样品中分离出一株病毒,该病毒能在MDBK细胞上增殖,能产生牛传染性鼻气管炎病毒(IBRV)典型性细胞病变。通过间接免疫荧光试验、微量血清中和试验、PCR及病毒形态学观察鉴定,证明该分离毒株为IBRV,并被命名为IBRV-LN01/08。根据IBRV的保守区核苷酸序列设计了1对鉴定引物,将扩增的目的片段进行克隆、测序及序列分析。结果表明,该片段序列与GenBank上登录的IBRV K22株的同源性为99.9%。动物回归试验结果显示,6~9月龄IBRV抗体阴性黄牛人工感染IBRV-LN01/08株F3代细胞培养物后,试验动物出现持续体温升高,流鼻涕,鼻内黏膜有白色糜烂溃疡及结节等牛传染性鼻气管炎的临床症状,表明该毒株具有一定的致病性。     

Dynamic changes of serum protein in rats with acute intoxication of Chinese cobra snake venom by proteomic analysis

To elucidate the toxic mechanism of snake venom at the protein level, proteomics technology was applied to investigate the effect of venom on circulation in the mammalian body. Temporal proteomic analysis was performed to profile the dynamic changes in the sera of Sprague–Dawley rats administered with Chinese cobra venom or saline. Using 8-plex iTRAQ analysis, 392 and 636 serum proteins were identified to be linearly upregulated or downregulated over time in the low-dose group and high-dose group, respectively. These proteins were mainly associated with the acute phase response pathway, complement system, and liver X receptor (LXR)/retinoid X receptor (RXR) and farnesoid X receptor (FXR)/RXR activation pathways. Compared with the low-dose group, the immune response and integrin pathways were inhibited in the high-dose group, although no obvious effect was observed. With consistently higher or lower expression in the high-dose group compared to the low-dose group throughout the whole process of venom poisoning, two proteins, Kininogen-1 (KNG1) and orosomucoid 1 (ORM1), which are involved in metabolism and immune response, occupied a core position in the pathway network and are considered venom dose-dependent biomarker candidates.     

猪源牛病毒性腹泻病毒的分离鉴定

从福建省某猪场疑似猪瘟的发病仔猪采集病料,处理后接种于牛肾细胞(MDBK),分离到1株病毒,并对其进行了系统鉴定.结果显示,病料接种MDBK细胞72 h后产生了明显的细胞病变;该分离毒株可以被牛病毒性腹泻病毒(BVDV)阳性血清中和,中和效价为1:106;该分离毒株对氯仿、乙醚、胰酶、酸敏感,不耐热,56℃30 min...     

不同培养系统牛体外受精囊胚的质量评价

采用双重荧光分染方法 ,结合免疫外科手术 ,对来自TCM 199和SOF培养系统的牛体外受精囊胚质量进行了评价。结果显示 ,TCM 199培养系统的囊胚发育率为 31.9% (335 / 10 74 ) ,平均细胞总数为 73.1,内胚团细胞数为 16 .8,其中生存细胞占 99.5 % ;SOF培养系统的囊胚发育率为 2 0 .9% (2 4 8/ 1186 ) ,平均细胞总数为 5 6 .2 ,内胚团细胞数为 11.3,其中生存细胞占 96 .5 %。证实 ,使用TCM 199生产的囊胚质量和囊胚发育率优于SOF培养系统。     

宁夏利通区乳牛育成期钙 磷盈亏状况调查

对宁夏吴忠市利通区 14个养牛场 (户 )进行了乳牛育成期钙、磷营养状况调查 ,测定了饲料钙、磷含量及育成期乳牛血中钙、磷、羟脯氨酸、碱性磷酸酶、甲状旁腺素、降钙素含量。结果表明 ,养牛小户的乳牛钙、磷代谢基本正常 ;散户、大户和中户的乳牛血钙不足 ;散户的乳牛血磷偏高 ,钙、磷比例失调。相关分析表明 ,钙与磷、甲状旁腺素呈中度正相关 ,相关系数分别为 0 .4 80、0 .4 5 0 ,钙与碱性磷酸酶呈中度负相关 ,相关系数为 - 0 .6 76 ;磷与羟脯氨酸呈中度正相关 ,相关系数为 0 .4 44。碱性磷酸酶与降钙素呈中度正相关 ,相关系数为 0 .4 85。降钙素处于正常范围 ,平均数为 4 .5 0 pmol/L± 6 .4 7pmol/L ,95 %的置信区间为 1.77～ 7.2 3pmol/L。     

猪胚胎干细胞培养和建系的影响因素及对策

为解决胚胎干细胞的培养和建系受血清、饲养层、细胞因子、传代时机以及其他一些不明因素影响而很不稳定的问题 ,在总结猪胚胎干细胞 (EG)培养和建系的经验教训的基础上 ,对一些常规的实验方法进行了改进 ,提出了消除上述影响因素的对策。     

神经生长因子对牛腔前卵泡体外培养的影响

用改良的McCoy’s 5a培养液(含 3 mmol/L L-谷氨酰胺、1 g/L BSA、29 ng/mL睾酮、2.5μg/mL转铁蛋白、100 ng/mL胰岛素和4 ng/mL硒),在96孔培养板中,每孔250μL培养液条件下,研究了神经生长因子(NGF)对牛腔前卵泡体外发育的影响。结果表明:在培养液中添加 3种不同浓度(4、6、8 IU/mL)NGF对体外培养的d<60μm腔前卵泡有促进生长的作用。在体外培养10 d时,3种不同浓度NGF组的d<60μm腔前卵泡及其卵母细胞直径平均增长值显著高于对照组,但对60μm130μm的腔前卵泡作用不显著。同时,不同浓度的 NGF显著提高了d<60μm腔前卵泡的发育率。     

Postmortem serum selenium concentrations and their possible etiological role in Sudden Infant Death (SID)

The importance of selenium (Se) deficiency in the pathogenesis of human diseases such as Keshan Disease has been extensively studied. It is possible that low Se-levels could cause immunosuppression and be an etiological factor in Sudden Infant Death (SID). We investigated 50 serum samples (40 SID and 10 non-SID victims) by atomic absorption spectrometry. The results show that there is no evidence of a serum selenium deficiency in SID-victims in the region of Aachen. A relationship between selenium concentration and the infant immune system still remains speculative and 21 samples even showed increased serum levels. It could be necessary to define a ‘local’ normal range by examining a greater number of healthy infants.     

牛病毒性腹泻病毒p80基因的分段表达及其抗原性分析

为建立以重组p80蛋白为抗原的牛病毒性腹泻(BVD)鉴别诊断ELISA方法,采用PCR方法克隆了BVDV p80基因的A、B、C三段基因片段,分别连接到原核表达载体pGEX-6P1上,获得了重组质粒pGEX-6P1-p80A、pGEX-6P1-p80B和pGEX-6P1-p80C,将其分别转化感受态菌Rosetta和BL21,经1.0 mmol/L的IPTG诱导,分别得到大小为60、47和51 ku的目的蛋白。经Western-blotting分析,3个目的蛋白中,只有p80B(289~477 aa)基因片段所表达的融合蛋白能与BVDV阳性血清反应,表明p80蛋白的免疫优势区域集中在此部位。该融合蛋白可以作为诊断抗原用于建立ELISA诊断方法。     

Postmortem Serum Tryptase Levels with Special Regard to Acute Cardiac Deaths

An elevated serum tryptase concentration is considered a specific marker for systemic mast cell activation, a central feature of anaphylaxis. However, in some cases of acute cardiovascular death, high concentrations of serum tryptase are also observed. We compared the postmortem serum tryptase concentrations in 74 cases assigned to the following four groups: anaphylactic deaths (Group A, n = 20), acute cardiac deaths (Group ACD, n = 30), acute dissecting aneurysm ruptures (Group ADA, n = 10), and controls (Group C, n = 14). Additionally, the cutoff between Group A and the other groups was calculated using receiver‐operating characteristic (ROC) curve analysis. Tryptase concentrations were markedly elevated in Group A (p < 0.001), Group ACD (p = 0.015), and Group ADA (p = 0.005). The optimal cutoff was 43 ng/mL, the sensitivity was 90%, and the specificity was 98%. While elevated concentrations of tryptase were noted in practical autopsy cases, due attention should be paid to the differential diagnosis between anaphylactic and acute cardiovascular deaths.