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.     

O型口蹄疫病毒VP1 T细胞与B细胞表位基因双拷贝串联表达产物的免疫应答

以一株O型口蹄疫病毒(FMDV)外壳蛋白VP1基因为模板,合成与细胞免疫及体液免疫相关抗原表位肽基因:21-40肽(20AA)和141-160肽(20AA)基因序列,运用基因工程技术构建了含有串联结构21-40(20AA)～141-160(20AA)～21-40(20AA)～141-160(20AA)的2020-2020VP1融合基因表达载体r2020-2020,转化宿主菌BL21(DE3)RIL后诱导表达,表达产物经SDS-PAGE及Western Blot分析显示重组融合蛋白的分子量约为18Ku.动物实验表明,较小剂量的融合蛋白就能诱导豚鼠产生特异性T淋巴细胞增殖反应及抗FMDV中和抗体,证明该融合蛋白可同时激活细胞免疫及体液免疫反应,具有开发成为抗FMDV疫苗的应用价值.     

流感病毒M2基因变异与盐酸金刚烷胺耐药性的关系

盐酸金刚烷胺是治疗流感的常用药物,但流感病毒极易产生耐药性并发生变异.本文仅就使用盐酸金刚烷胺后流感病毒M2基因的变异情况作一综述.     

转基因植物疫苗研究进展

众多研究表明,病毒的抗原决定簇及细菌毒素亚单位均能在转基因植物中成功表达,并保持良好的免疫原性.与现有的疫苗生产体系相比,植物生产疫苗具有安全、经济、稳定、高效等优势.本文概述了利用不同受体植物生产疫苗的研究现状、免疫原性评价及免疫原基因的优化表达策略.     

复方磷霉素钙水溶性粉处方优化实验

为了优选磷霉素钙复方制剂,采用体外药物最小抑菌浓度测定法,测定了磷霉素钙与甲氧啶(TMP)、丁胺卡那霉素、恩诺沙星、庆大霉素、头孢噻肟钠、多西环素、氟甲砜霉素联合使用对大肠杆菌作用的协同指数.结果表明磷霉素钙与TMP的协同关系最好,协同指数为0.281,制剂中磷霉素钙与TMP的含量分别为5%和1%为最佳.     

乌兰布和沙漠流动沙丘不同部位水分动态研究

通过对乌兰布和沙漠流动沙丘的不同部位沙土含水量的测定研究，结果表明：（1）乌兰布和沙漠流动沙丘3个不同部位的水分垂直变化趋势是一致的，0～60cm含水量变化幅度大，60～100cm变化幅度小，含水量最大值出现在20～60CITI，降雨主要影响60cm以上的沙土水分；（2）3种部位在时间变化上都表现为4～5月含水量最小，从6月开始含水量升高；（3）迎风坡含水量较其他部位大0．65％～0．9％，较背风坡大1．8％～2．5％，其他部位较背风坡大1％～1．7％，且3种地貌部位20～60cm处较80cm和100cm处有较大差异，同时迎风坡的变化幅度最大。     

广东鹤山、东莞莲花山小蛾类（Ⅰ）

对广东省鹤山和东莞莲花山自然保护区的小蛾类昆虫进行调查，计有8科35属43种。其中广东新记录种14种。     

A COMPARISON OF THE ROSTROCAUDAL OPEN MOUTH AND ROSTRO 10° VENTRO-CAUDODORSAL OBLIQUE RADIOGRAPHIC VIEWS FOR IMAGING FLUID IN THE FELINE TYMPANIC BULLA

Fluid within the tympanic bulla is an indication of middle ear disease. Radiography has a relatively low accuracy for the detection of soft tissue opacification in the tympanic bulla, and the most useful radiographic projection, the rostrocaudal open mouth (RCd (open mouth)), is technically difficult to perform in dogs and cats. An alternative projection for the feline tympanic bulla, the rostro 10 degrees ventro-caudodorsal oblique (R10 degrees V-CdDO), was compared to the RCd (open mouth) in 41 feline cadaver heads with the tympanic bullae randomly filled with KY jelly. Computed tomography was used as the gold standard. Each tympanic bulla was recorded as being positive or negative for soft tissue opacification. Although there was no significant difference between the accuracy of the two views, the R10 degrees V-CdDO was subjectively more accurate and easier to perform, and in a live patient may be performed without the need for general anesthesia. The R10 degrees V-CdDO projection is a good alternative to the RCd (open mouth) projection for detecting otitis media in the cat.     

ECHOCARDIOGRAPHIC AND DOPPLER DIAGNOSIS: FIRST CASE OF ATRIAL SEPTAL ANEURYSM IN A CAT

A 6-year-old female cat developed pleural and abdominal effusion. Cardiac ultrasound and 2D color tissue Doppler imaging revealed restrictive cardiomyopathy with severe biatrial dilatation and hypertension. This cardiomyopathy was associated with atrial septal aneurysm and a patent foramen ovale. The atrial septal aneurysm involved the entire atrial septum. Necropsy and histological examination confirmed all these findings. ASA is a rare malformation and, as in this cat, it generally occurs concomitantly with congenital or acquired heart disease and may be explained by greater pressure in one atrium, leading to controlateral protrusion of the atrial septum.     

CONTRAST-ENHANCED ULTRASONOGRAPHY IN NORMAL CANINE LIVER. EVALUATION OF IMAGING AND SAFETY PARAMETERS

Contrast-enhanced ultrasonography, a new imaging modality in veterinary medicine, can provide data on tissue perfusion. The objective of this study was to use the ultrasonographic contrast agent SonoVue to evaluate various transit time indices in the normal canine liver, to examine the effect of anesthesia on these parameters, and to evaluate the safety of this agent in dogs. The liver of 11 healthy dogs was studied by ultrasound during an intravenous bolus injection of SonoVue. Each dog underwent the examination twice, first with and later without the use of anesthesia. A time-intensity curve was generated from a selected region of interest within the liver from each scanning session. Ratios derived from peak enhancement, time to peak enhancement, up-slope and full-width half-maximum (FWHM) of the curve were calculated from the time-intensity curves, and are reported. There were no statistically significant differences (P > 0.05) in peak enhancement, up-slope and FWHM between dogs that were anesthetized and dogs that were not. Time to peak enhancement, however, was significantly shorter when the dogs were anesthetized than when they were nonanesthetized (P < 0.05). There were no biologically significant changes in clinical laboratory findings. This study indicates that contrast-enhanced ultrasound using SonoVue gives reproducible liver perfusion data, and appears to be a safe and well-tolerated agent for use in dogs. When considering normal values, the use of anesthetic drugs has to be considered.