牛分枝杆菌MPB70基因的克隆及其在大肠杆菌中的表达

本研究以牛分枝杆菌Vallee111染色体DNA为模板,以MPB70成熟蛋白基因特异性引物进行PCR扩增,获得约500bp的DNA片段.通过T-A克隆技术,将PCR产物克隆至pGEM-T Vector中,成功地构建出克隆载体pGEM-T-70.以BamH Ⅰ和EcoR Ⅰ双酶切pGEM-T-70和pET28a( ),并将纯化的MPB70基因亚克隆至pET28a( )中,构建出原核表达载体pET28a-70.将pET28a-70转化至感受态E.coli BL21(DE3)中,经IPTG诱导和SDS-PAGE分析,可见约25Ku外源蛋白带.Western blot分析发现,该蛋白具有牛分枝杆菌抗原性,从而为进一步研究MPB70的亚单位疫苗及DNA疫苗奠定基础.     

广西猪瘟流行毒株生长特性的初步研究

本实验以来自广西梧州、博自和北海地区的3株猪瘟野毒和中国标准石门株病毒、疫苗毒为材料,研究其在宿主体内和体外PK-15细胞中增殖的基本特性和差异.结果显示,广西的一部分毒株很可能在毒力和生长特性上已经发生了一些变异.本实验用免疫酶实验检测PK-15细胞中的猪瘟病毒抗原,发现其主要存在于细胞的核膜以及内质网和高尔基体富集的区域.     

油乳剂疫苗中硬脂酸铝含量与油相水相比例关系研究

进行了鸡新城疫、传染性支气管炎、产蛋下降综合征三联油乳剂疫苗生产中降低油相比例的试验.当油乳剂疫苗的油相中硬脂酸铝含量分别为2%、1%、0.5%,对应的油相:水相分别为3:1、2:1和1:1时进行的乳化试验结果表明:随着油相中硬脂酸铝含量降低,油相和水相的比例相应降低,呈正比例关系.当油相中硬脂酸铝含量与油/水比分别为2%和3:1、1%和2:1、0.5%和1:1时,制备的油苗质量均达到了国家标准.     

复方罗红霉素可溶性粉对鸡葡萄球菌病的疗效试验

采用人工诱发的病理模型,按推荐剂量治疗,验证复方罗红霉素可溶性粉对鸡(试验时30日龄)葡萄球菌病的疗效.试验结果表明,复方罗红霉素可溶性粉对鸡葡萄球菌病有很好的治疗效果,有效率达96.7%;推荐剂量治疗组的死亡率极显著地低于感染对照组(P<0.01);推荐剂量治疗组与健康对照组的增重差异不显著(P>0.05),但极显著高于感染对照组(P<0.01).     

疑似碘酊致牛死亡一例

笔者在临床上曾遇到一疑似“碘酊”致牛死亡的病例,由于未曾见过碘酊致牛死亡的报道,查阅资料也终未得到答案.笔者将其整理如下,供同行探讨。     

Effect of oxymatrine on mouse allergic contact dermatitis induced by DNFB and lymphocyte proliferation stimulated by Con A

AIM: To explore the inhibitory effect of oxymatrine （OMT） on the allergic contact dermatitis （ACD） stimulated by dinitrofluorobenzene （DNFB） and its effects on the proliferation of the lymphocytes. METHODS: ① An ACD mouse model was established by stimulation with DNFB, and then the mice were injected intraperitoneally with different dosages of OMT， PBS and hydrocortisone （HCT） respectively, the swelling degree of their auricles was examined. ② Carboxyfluorescein diacetate, succinimidyl ester (CFDA-SE) dye and flow cytometer were used to examine the fluorescence intensity changes of lymphocytes stimulated by polyclonal stimulator ConA and OMT. RESULTS: ① compared with PBS group, OMT possessed the strong inhibitory effect on the ACD caused by DNFB in a dose-dependent manner, and its inhibitory effect was equivalent to the HCT of the same dosage with fewer side effects. ② In vitro experiments proved that OMT (500, 125 and 31 mg/L) had the ability to restrain the proliferation of lymphocytes of mouse. CONCLUSION: OMT possesses an inhibitory effect on the ACD induced by DNFB, and OMT is a kind of immunosuppressor.     

Macaca irus bone mesenchymal stem cells differentiate into neuron-like cells induced by cryptotanshinone in vitro

AIM: To determine the optimal protocol and condition in which macaca irus mesenchymal stem cells (MSCs) are induced to differentiate into neuron-like cells by cryptotanshinone in vitro. METHODS: MSCs from macaca irus bone marrow were generated in vitro and induced with cryptotanshinone. The morphological changes of MSCs were evaluated by microscope. The positive percentages of neurofilament (NF), neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression were measured by immunocytochemistry with ABC staining. RESULTS: The result showed that MSCs were positive for CD29, CD44, CD105, CD166, and negative for CD34, CD71, CD80 and CD86. After induced with cryptotanshinone, MSCs began to display neuronal morphologies, such as contracted multipolar cell body and formed extensive networks. The percentages of positive NSE, NF expression were 68.3%±3.5%, 70.3%±1.5%， respectively. CONCLUSION: Macaca irus MSCs could be induced to differentiate into neuron-like cells in vitro by cryptotanshinone and might be applied in cell transplantation and gene therapy in nervous system disorders.     

毛叶枣病害调查及炭疽病的研究进展

本文对云南省毛叶枣的炭疽病、焦腐病、白粉病、疫病、黑斑病等真菌性病害和缺硼、缺镁及缺铁的发生情况作了介绍，并简单地概述了炭疽病的研究进展。