鸡传染性支气管炎病毒(IBV)ZZ2004株核衣壳蛋白基因的克隆及原核表达

本研究旨在克隆并表达鸡传染性支气管炎病毒（IBV）的核衣壳蛋白（N）。根据IBV ZZ2004株的N基因设计1对引物,提取IBV ZZ2004株的RNA,利用RT-PCR技术扩增大小约1.23 kb的片段,将其插入克隆载体pGEM-T构建pGEM-T-N。将pGEM-T-N用EcoRⅠ和XhoⅠ进行双酶切,将酶切后的N基因片段克隆到表达载体pGEX-6P-1,转入大肠杆菌BL21（DE3）菌中,经IPTG诱导表达,进行SDS-PAGE电泳,结果显示外源基因获得了表达且为可溶性蛋白;Western blotting检测结果显示N蛋白可与相应的IBV ZZ2004阳性血清发生特异性反应,结果表明N蛋白有一定的生物活性。本研究为IBV诊断试剂盒及新型IBV重组亚单位疫苗研制奠定基础。     

猪蛋白质二硫化物异构酶A4基因克隆及密码子偏好性分析

试验旨在研究猪蛋白质二硫化物异构酶(protein disulfide isomerase A4,PDIA4)基因的密码子使用特性及其与其他物种密码子使用的差异。试验以香猪子宫组织为试验材料,采用RT-PCR方法扩增获得猪PDIA4基因,参考从NCBI数据库中下载的18个不同物种PDIA4基因序列,对猪PDIA4基因的密码子使用特性及其与其他物种密码子的使用差异和进化关系进行分析,同时对猪PDIA4基因与4个模式生物密码子使用偏性进行比较。结果表明,猪PDIA4基因编码区长1 941 bp,共编码646个氨基酸。与NCBI数据库公布的野猪PDIA4基因(GenBank登录号:NM_001267834.1)的相似性达99%。猪PDIA4基因偏好使用G/C结尾的密码子,27种密码子为猪PDIA4基因的偏好密码子(RSCU>1),其中使用频率最高的前3个密码子分别为CTG(RSCU=3.69)、GCC(RSCU=2.31)和CGC(RSCU=2.29)。比较19个物种PDIA4基因发现,不同物种间PDIA4基因的有效密码子数(ENc)、密码子适应指数(CAI)、GC含量以及T、C、A、G、G+C在密码子第3位含量(T3s、C3s、A3s、G3s、GC3s)均存在差异,且密码子偏好以G/C结尾;聚类分析发现,基于CDS序列的系统进化树更符合19个物种的真实分类系统。在密码子使用频率上,猪PDIA4基因与小鼠基因组间密码子使用偏好程度更为接近,故小鼠更适合作为猪PDIA4基因的遗传转化和异源表达的受体系统。     

气候变化对藏药独一味适生区分布格局的影响

本研究通过实地调查,运用最大熵(MaxEnt)模型和环境气候数据模拟未来气候变化对青藏高原地区藏药独一味(Lamiophlomis rotata(Benth.)Kudo)适生区空间分布格局的影响,探索其适生区变迁规律.通过ArcGIS和SPSS对模拟结果分析显示,当前气候条件下独一味最适宜生长地区主要分布于青藏高原地区...     

Cell-based cancer gene therapy: breaking tolerance or inducing autoimmunity?

This review examines the mechanisms involved in anti-tumor immunity and how peptides present in many tumor types (tumor-associated antigens) are recognized by T cells from tumor-bearing cancer patients. Tumor-associated antigens are derived from proteins that are also expressed in normal cells. It is predicted that immune responses to such peptides will be compromised by self-tolerance or that stimulation of effective immune responses will be accompanied by autoimmunity. We also consider that the immunity induced against two autoantigens, which are highly conserved in vertebrates, involve qualitatively different mechanisms, such as the production of antibodies and cell-mediated immune responses. However, both pathways lead to tumor immunity and identical phenotypic manifestations of autoimmunity. Appropriate selection of the optimal tumor antigen is critical for the induction of an anti-tumor immune response. Thus, we stress that the methods for antigen presentation using dendritic cells play a critical role in the development of tumor vaccines, to break immune tolerance and induce a strong immune response against them. The viability and feasibility of expansion of canine dendritic cells from bone marrow and peripheral blood ex vivo for the treatment of spontaneous cancers in dogs is also discussed.     

Identification of RSVP14 and RSVP20 components by two-dimensional electrophoresis and Western-blotting.

We have already shown that RSVP14 and RSVP20, two ram seminal plasma (SP) proteins postulated to be involved in sperm capacitation and gamete interaction can protect spermatozoa against cold-shock. In this study, we use two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) for the analysis of SP proteins of Rasa Aragonesa rams, using enhanced protein solubilization in the presence of tributyl phosphine (TBP) and a polyacrylamide linear gradient gel with a narrow pH range (4-7). The image analysis of the 2D map detected 195 protein spots, with isoelectric points (pIs) ranging from 4.5 to 6.6, and molecular weight (M(r)) from 11.7 to 90.4. Staining of 2D gels with Pro-Q Emerald 300 Glycoprotein Stain revealed that most significant proteins in ram SP are glycosylated. The removing of protein N-linked oligosaccharides improved the gel resolution. 2D-PAGE analysis of the whole fraction 6 (F6) separated from ram SP by exclusion chromatography showed six main protein spots, four (a, b, c, d) in the 14 kDa and two (e, f) in the 20 kDa region. Western-blot analyses indicated that the anti-P14 antibody recognized four spots on the SP map, 4, 5, 6 and 7, that matched with spots a, b, c, d of F6 map. The anti-P20 antibody recognized spots 13 and 14 of SP map that corresponded to spots e, f of F6 map. The deduced sequences by de novo sequencing evidenced that protein spots 7 and 13 have significant similarities to BSP family, while protein spots 4 and 14 did not appear to be homologous with any reported protein in the current mammalian Proteinbank databases.     

夏季日粮中主要营养水平对肉种母鸡生产性能的影响

为研究夏季日粮中粗脂肪、粗蛋白、赖氨酸和蛋氨酸水平对肉种鸡生产性能的影响,27周龄良凤肉用种母鸡随机分为4个处理,每个处理3个重复,每个重复为490只,处理一饲喂对照日粮(CF2.7%,CP16.5%,Lys0.83%,Met0.40%),处理二饲喂高脂肪日粮(CF5.43%),处理三饲喂高蛋白日粮(CP18.2%),处理四饲喂高赖氨酸和蛋氨酸日粮(Lys0.91%,Met0.44%),试验期共112天。结果表明,对照日粮的营养水平是适合的,在此基础上分别增加日粮的粗脂肪水平、粗蛋白水平、蛋氨酸和赖氨酸水平,都不能提高肉种母鸡的生产性能,也不能减轻夏季炎热带来的影响。     

Effects of 5-lipoxygenase inhibitor zileuton on destruction of blood-brain barrier permeability induced by focal cerebral ischemia-reperfusion injury in rats

AIM: To investigate the influential factors of the blood-brain barrier (BBB) permeability and to observe the effects of zileuton, a selective 5-lipoxygenase inhibitor (5-LO), on focal cerebral ischemia-reperfusion injury (CIRI).METHODS: The right middle cerebral artery of the rat was occluded by inserting a thread through internal carotid artery for 2 h, and then reperfused for 24 h. Zileuton (10, 50 mg·kg-1, po) was orally administered 2 h before ischemia and at 0, 5, 10 h after reperfusion. The permeability of blood brain barrier (BBB) was detected by using Evans blue (EB) as a labelling compound. The degree of cerebral edema was estimated by AutoCAD image analysis software. The mRNA of cysteiny leukotrienes receptor1 (CysLTR1) was detected by RT-PCR. The content of LTB4 in serum was measured by enzyme linked immunosorbent assay (ELISA). The expressions of AQP4 and MMP-9 proteins were measured by immunohistochemical staining method. RESULTS: After middle cerebral artery occlussion 2 h/reperfusion 24 h, the permeability of BBB in the brain tissue of injured side and the brain edema degree were increased. The content of LTB4 in serum was elevated. The expression of CysLTR1 mRNA from the brain tissue of occluded side was enhanced. The expressions of MMP-9 and AQP4 proteins of the ischemia realm and ischemia penumbra (IP) of the infarct focus perimeter were increased. Both 10 and 50 mg·kg-1 doses of zileuton dramatically relieved the BBB permeability destruction and the degree of the brain edema, inhibited the expression of CysLTR1 mRNA in the brain tissue and also reduced the content of LTB4 in serum. The expressions of AQP4 and MMP-9 proteins in the brain tissue were also decreased.CONCLUSION: The permeability of BBB is destroyed after the focal CIRI. The mechanisms of protective effect of zileuton might be attributed to its effects by inhibiting the activation of 5-LO pathways on the brain tissue and circulatory blood, reducing the expressions of AQP4 and MMP-9 proteins of the ischemia and IP realm in the brain tissue.     

RADIOGRAPHIC FINDINGS IN DOGS WITH SPONTANEOUS PULMONARY THROMBOSIS OR EMBOLISM

Thoracic radiographs of 21 dogs with necropsy-confirmed pulmonary thrombosis or embolism (PTE) were reviewed. Two major lung patterns were seen: (1) regional oligemia, giving the impression of a hyperlucent zone; (2) alveolar pulmonary infiltrate, indicating hemorrhage or infarction. Hemorrhage and infarction could not be differentiated radiographically. Other common findings were attenuation or loss of visualization of a lobar artery or lobar vein and pleural effusion, particularly with infarction and cardiac disease. PTE was often associated with renal disease (glomerulonephritis, amyloidosis), septic processes, pancreatitis, systemic venous or right heart thrombosis, cardiac insufficiency, and hypercorticism.     

达乌里胡枝子花粉萌发特性研究

为揭示达乌里胡枝子(Lespedeza daurica)花粉的育性并为其花粉活力检测提供一种高效方法,本试验以达乌里胡枝子新鲜花粉为试验材料,采用液体培养基,研究了蔗糖、硼酸、氯化钙浓度、采样时间、温度、光照和培养时间等因素对花粉离体萌发和花粉管生长的影响.试验结果表明:单因素试验中,适宜花粉萌发的培养基配方为20 g...