云南地区部分种猪场种公猪疫病的调查与分析

生产中,种公猪多种疫病会通过水平或垂直传播给种母猪或仔猪,造成种猪群繁殖障碍和呼吸道疾病暴发.为了解云南地区规模种猪场公猪疫病的感染情况,于2014年对云南10个地区63个种猪场4个品种的101头公猪血液和78头公猪精液进行了相关血清学和抗原检测.血清学检测结果显示:公猪血液中含有猪支原体、猪布氏杆菌病、猪弓形体病、猪传染性胸膜肺炎的抗体,其比例分别为32.67%、6.93%、8.91%、28.71%;其中,单独感染某一疫病30头占29.7%,二重混合感染有21头20.8%,三重混合感染2头1.98%,未见四重混合感染.对公猪精液进行PRV、CSFV、PRRSV、PCV-2、PPV和JEV的PCR检测发现:PRRSV感染检出率为12.82%,CSFV+PRRSV2二重混合感染检出率3.85%,PRRSV+PCV-2+JEV三重混合感染检出率2.56%.通过血清学和病原学共检测的179头种公猪,75头单独或混合感染以上10种病原,感染率42.13%,阳性猪场52家,占82.53%.     

Analysis of the distribution and clonality of TCR Vβ T cells in cord blood

AIM:To investigate the distribution and clonality of TCR Vβ subfamily T cells in cord blood. METHODS:The CDR3 of TCR Vβ 24 subfamily genes were amplified in mononuclear cells from 13 cases of cord blood. To observe the usage of TCR Vβ repertoire, the PCR products were further labeled with fluorescent and analyzed by genescan technique for the CDR3 size, to evaluate clonality of the detectable TCR Vβ T cells. Peripheral bloods from 10 cases of normal individuals and T cell line Molt-4 and Jurkat served as controls. RESULTS:Only 38.78%±16.26% of 24 Vβ subfamily T cell were selectively expressed in cord blood, predominantly in Vβ 3, 5, 8, 9 and 13, whereas all 24 Vβ subfamilies could be detected in T cells from peripheral blood of normal individuals. Genescan analysis showed that all PCR products of TCR Vβ subfamilies from cord blood or normal individual peripheral blood displayed multi-peaks. CONCLUSION:Some TCR Vβ subfamily T cells were absent in cord blood. All TCR Vβ subfamily T cells in cord blood displayed polyclonality.     

Preparation and Preliminary Application of Anti-cephalexin Monoclonal Antibody

The aim of the study was to prepare a monoclonal antibody (McAb) against cephalexin(CEX),and preliminarily establish an ELISA method to detect residues of CEX in milk. The CEX was conjugated to carrier bovine serum albumin (BSA) and ovalbumin (OVA) by two-step glutaraldehyde method. BALB/c mice were immunized with the prepared CEX-BSA conjugate. After five times immunization, we built and screened two strains of hybridoma cells (2G4 and 5B3) which secreted McAb against CEX by hybridoma technology.The 2G4 cell lines were injected into BALB/c mouse's abdominal cavity to produce ascites. Finally the ascites were purified and identified. The results showed that antibody titer of the McAb 2G4 was above 1∶1.28×10⁵,its antibody subtype was IgG2b(κ) and affinity constant K was 1.51×10⁹ L/mol. The cross experiment result evidenced that the cross reaction rate of cefradine was 52.55%, ceftiofur, ceftriaxone, cefotaxime, cephalothin, penicillin, streptomycin and tetracycline had no cross reaction. We established an ELISA method to detect CEX residues in milk and its linear range was 20 to 1 000 ng/mL, linear equation y=0.4674x-0.5359(R²=0.9909), its limit of detection (LOD) was 19.68 ng/mL and the average recovery rate was 91.31%, the average coefficient of variation was below 15%. The detection limit was lower than the maximum residue limit of cefalexin in China and European Union. So this detection method might have good application foreground.     

牛皮蝇蛆病间接ELISA诊断方法的建立

以纹皮蝇Ⅰ期幼虫粗提蛋白为包被抗原,通过方阵试验确定了血清最佳稀释倍数为80倍,抗原最佳包被浓度为26.64μg/mL,并对其特异性、敏感性和重复性进行了试验,建立了牛皮蝇蛆病间接ELSIA诊断方法。再以建立的ELISA诊断方法对采自内蒙古地区的233份牛血清进行了检测。结果表明,所建立的牛皮蝇蛆病间接ELISA诊断方法具有较好的特异性、敏感性和可重复性,可用于牛皮蝇蛆病的血清学检测。     

酶标抗原Dot－ELISA检测鸭肝炎病毒抗体

酶标抗原Dot－ELISA检测鸭肝炎病毒抗体@杨奎@陈溥言...     

Inhibitory effect of berberine on the activation and proliferation of T lymphocytes

AIM: To investigate the effect of berberine (Ber) on the activation and proliferation of T lymphocytes and its mechanism of action. METHODS: Whole peripheral blood from normal subjects was stimulated with phytohemagglutinin (PHA) or phorbol ester (PDB) plus ionomycin (Ion) and the expression levels of CD69 and CD25 were evaluated with flow cytometry after the staining with appropriate fluorescent monoclonal antibody. The distribution of cell cycles was analyzed by propidium iodide staining and dead cells by 7-aminoactinomycin live staining. RESULTS: 100 μmol/L and 50 μmol/L of Ber had significant inhibition of the expression of CD69 on T cells stimulated with PDB plus Ion or PHA, while effect of 25 μmol/L Ber was not significant. And as time of action extended, the extent of inhibition decreased. For the expression of CD25, Ber at the concentrations as above all exerted significant inhibitory effect in a dose-dependent manner. Moreover, Ber could block lymphocytes cell cycle progression from G₀/G₁ phase to S and G₂/M phase without phase specificity. Besides, live staining analysis revealed that Ber did not have significant cytotoxicity on lymphocytes. CONCLUSIONS: Ber significantly inhibits the expression of early and mid activation antigens of T cells and also blocks the progression of lymphocytes cell cycles. These results suggest that Ber exerts immunosuppression effect through inhibiting the activation and proliferation of T cells.     

H5亚型禽流感病毒血凝素基因的克隆及表达

研究以RT-PCR方法从H5N1亚型禽流感病毒A/DKZJ/12/00(H5N1)中获得禽流感病毒HA部分基因,将目的基因定向克隆到原核表达载体pET-32a,将测序和酶切验证正确的阳性重组质粒转化大肠杆菌BL21,经IPTG诱导表达。结果表明该蛋白得到了可溶性的表达,表达蛋白的分子质量为25ku;Western-blot分析结果表明,该蛋白可以与禽流感H5阳性血清反应(禽流感的单克隆抗体所识别),检测HA的抗体时具有良好的免疫原性。     

马立克氏病病毒(MDV)糖蛋白gB在重组鸡痘病毒中的表达

将马立克氏病病毒gB基因用EcoR I从质粒pUR-MDVEgB中切下。两端补平后，插入到质粒pEFgpt12s的Pst I酶切位点。构建了含有完整的MDVgB基因的转移载体pEFgpt12s-gB。这个载体的特点是，它含有两个启动子，在强的复合启动子Spromoter的下游是插入的gB基因，另一个反向启动子vvP7.5的下游 是标记基因Ecogpt，它的两端是FPV的非必需区。携带gB基因的质粒pEFgpt12s-gB通过磷酸钙的方法转染用282e4株FPV感染3-4h的鸡胚成纤维细胞。通过药物筛选 ，得到含有gB基因的重组病毒。通过PCR、免疫荧光检测，证实了重组病毒中含有gB基因，并且gB糖蛋白也得到了表达。     

Salmonella abortus ovis (Etude de 112 souches)

The bacteriological examination of 112 strains of Salmonella abortus ovis shows some particular properties of this serotype, and the existence of two different types according to their geographical source.