Characterization of bovine ileal epithelial cell line for lectin binding,susceptibility to enteric pathogens,and TLR mediated immune responses

In this study, primary and immortalized bovine intestinal epithelial cells (BIECs) were characterized for the expression of surface carbohydrate moieties. Primary BIEC-c4 cells showed staining greater than 90 % for 16 lectins but less than 50 % staining for four lectins. Immortalized BIECs showed significantly different lectin binding profile for few lectins compared to BIEC-c4 cells. BIEC-c4 cells were studied for infectivity to E. coli, Salmonella enterica, bovine rotavirus, bovine coronavirus, and bovine viral diarrhea virus. Bovine strain E. coli B41 adhered to BIEC-c4 cells and Salmonella strains S. Dublin and S. Mbandaka showed strong cell invasion. BIEC-c4 cells were susceptible to bovine rotavirus. LPS stimulation upregulated IL-10, IL-8, and IL-6 expression and Poly I:C upregulated TLR 8 and TLR 9 expression. This study provides important knowledge on the glycoconjugate expression profile of primary and immortalized BIECs and infectivity and immune responses of primary BIECs to bacterial and viral pathogens or ligands.     

鸡源株与人源株H9N2流感病毒血凝素受体结合位点氨基酸比较与分析

H19N2病毒可以感染多种禽类和哺乳动物，包括人类。流感病毒血凝素受体结合位点的氨基酸可以影响受体结合特性。为了解鸡源株与人源株H19N2病毒受体结合部氨基酸的差异，作者对二者进行了比较与分析，结果表明，鸡源株与人源株血凝素受体结合部位氨基酸的第137、183、190、226位点存在差异，鸡源株在这些位点分别为K、N、AorVorT、LorQ而人源株则分别为R、H、E、L。虽然尚不清楚这些位点的改变对病毒与细胞亲和力及宿主范围的影响，但由于H9N2在我国养鸡业的普遍存在，加强H9N2病毒的分子流行病学监测有重要意义。     

棉花黄萎病菌毒素结合位点初探

以纯化的棉花黄萎病菌毒素(PLPC)免疫新西兰白兔制备了PLPC特异性抗血清。将PLPC(15μg·ml 1)用不同浓度的抗体(1～20μg·ml 1IgG)吸附后处理泗棉3号的切根苗,毒素所引起的症状都有不同程度的减轻。表明所制备的抗体在与毒素发生特异性免疫学反应的同时,可部分封闭毒素分子上与毒素受体结合的位点。利用竞争ELISA测定了泗棉3号幼苗子叶的质膜制剂与PLPC的结合活性。结果表明,质膜制剂与毒素结合后能部分阻断毒素与其抗体的免疫学反应,即质膜制剂中含有毒素的结合位点。分别用胰蛋白酶和煮沸处理质膜制剂后,质膜制剂对毒素与其抗体的反应的抑制作用消失,初步表明质膜制剂中与毒素结合的是蛋白质。     

淡紫拟青霉发酵滤液对大豆胞囊线虫趋化性的影响

 实验室条件下,研究了大豆胞囊线虫幼虫对大豆根和根浸出液的趋化性以及淡紫拟青霉代谢产物对其趋化性的影响。结果表明,大豆胞囊线虫二龄幼虫(J₂)对大豆根存在着一定的趋化性,而淡紫拟青霉发酵滤液对J₂存在明显的驱避性。在根浸出液、真菌滤液及其混合液剂处理下,大豆胞囊线虫J₂在WA平板上靠近处理液的0~1cm区间中分布率存在极显著差异(P <0.0 1),而加样时间(液剂与线虫同时放置或提前2 4h处理)对其分布率没有显著影响。大豆幼根蘸取滤液后对线虫J₂在该区间的分布与不经处理的幼根差异显著(P <0.0 5),证明淡紫拟青霉发酵滤液可以明显地降低线虫与大豆根的亲和力,并强烈抑制线虫对大豆根的侵染。     

西瓜抗枯萎病基因同源序列的克隆与分析

本研究根据已克隆的抗枯萎病基因的NBS保守结构域设计了22条上游简并引物和17条下游简并引物,以西瓜抗枯萎病种质PI296341-FR和感枯萎病品种97103为材料,获得了7条来自基因组DNA的RGA序列（GenBank登录号：DQ156558-DQ156564）,均含有NBS保守区的P-环、kinase-2或kinase-3等抗病基因的特征序列结构,所编码的氨基酸序列与已知抗枯萎病基因Fom-2、I2C-1、I2C-2和I2等编码的氨基酸序列表现出11%～72%的同源性,其中来自PI296341-FR的RGA序列175R1与甜瓜抗枯萎病基因Fom-2的同源性最高,为72%。来自PI296341-FR与97103的RGA序列之间同源性较高（73%～97%）,证明了抗病基因在进化上的保守性。     

牙鲆血清免疫球蛋白的分离纯化及部分特性分析

运用Sephacryl S-200凝胶层析和HiTrap rProtein ASepharose亲和层析2种方法对牙鲆(Paralichthys olivaceus)血清免疫球蛋白进行分离纯化,结果表明,牙鲆免疫球蛋白分布于33%~50%的硫酸铵饱和溶液中,其中45%的分离效果最好。凝胶层析和亲和层析样品均出现2个蛋白峰,用还原SDS-PAGE检测确定牙鲆免疫球蛋白存在于第2个蛋白峰中。牙鲆免疫球蛋白重链分子量约为75.4 kD,轻链分子量约为29.9 kD和28.2 kD,推测牙鲆血清免疫球蛋白的分子量为836 kD。制备了兔抗牙鲆免疫球蛋白多克隆抗体,免疫双扩散法检测多克隆抗体效价为1∶32,免疫斑点法检测多克隆抗体效价至少为1∶1 600。运用免疫印迹法(Western-bloting)检测了兔抗牙鲆免疫球蛋白多克隆抗体的特异性,实验证明该抗体与牙鲆全血清中免疫球蛋白重链、轻链反应均成阳性。     

图书排版装帧对新书加工细节的影响

分析现代图书排版装帧的现状与对图书馆新书加工一些细节的影响,对图书的排版装帧设计提出一些设想     

Effect of miRNA-33 on ABCA1/ABCG1 expression in RAW264.7 macrophage-derived foam cells after Hcy treatment

AIM:To study whether homocysteine (Hcy) inhibits the expression of ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) by microRNA-33 (miRNA-33) signaling, and reduces the efficiency of reverse cholesterol transport (RCT).METHODS:RAW264.7 macrophages were induced by oxidized low-density lipoprotein (ox-LDL) to establish foam cell model. Oil red O staining was used to determine whether the model was established successfully. miRNA-33 mimics and miRNA-33 inhibitor were transfected into the cells by Lipofectamine 2000, and the cells were exposed to Hcy at concentration of 5 mmol/L for 24 h. The intracellular lipid droplets were observed by Oil red O staining. The expression of ABCA1 and ABCG1 at mRNA and protein levels was determined by real-time PCR and Western blot. The cellular cholesterol content was analyzed by HPLC, and effluent rate of cholesterol was detected by the method of liquid scintillation counting.RESULTS:Compared with blank control group, the lipid content in miRNA-33 mimics group was increased, and the expression of ABCA1 and ABCG1 at mRNA and protein levels was decreased (P<0.05). The intracellular cholesterol content was increased gradually (P<0.05), and the cellular cholesterol efflux rate was gradually decreased (P<0.05) in miRNA-33 mimics group. Compared with blank control group, the testing results in miRNA-33 inhibitor group were the opposition of those in miRNA-33 mimics group (P<0.05). No diffe-rence of the above indexes among blank control group, miRNA-33 mimics-NC group and miRNA-33 inhibitor-NC group was observed.CONCLUSION:Hcy inhibits the mRNA and protein expression of ABCA1 and ABCG1 through miRNA-33 signaling, and reduces the efficiency of RCT in RAW264.7 macrophage-derived foam cells.     

华北大黑鳃金龟气味结合蛋白HoblOBP2的生物信息学分析

探讨应用生物信息学技术预测华北大黑鳃金龟触角气味结合蛋白HoblOBP2三维结构及其与苯甲酸己酯的结合模式.利用生物信息学相关工具和网站预测HoblOBP2的基本性质,并对其三维结构进行同源建模后发现HoblOBP2的三维结构具有与昆虫普通气味结合蛋白结构的共同特点,即6个α螺旋,其中除了α2在外的5个α螺旋形成疏水结合口袋,3对二硫键起着稳定蛋白结构的作用.将苯甲酸己酯分子对接到HoblOBP2的活性区域,预测蛋白质与气味分子的结合模式,经分析可知蛋白质与气味分子主要通过范德华力和疏水作用相互影响,蛋白内部形成分子内氢键,根据蛋白与气味分子间的结合特性推测HoblOBP2与气味分子结合的三个关键结合位点分别为:LEU73,THR54和VAL137.     

斜纹夜蛾化学感受蛋白SlitCSP2的表达模式定位及配体结合特性分析

研究斜纹夜蛾Spodoptera litura化学感受蛋白Slit CSP2的表达模式、组织定位及与配体结合特性,为揭示SlitCSP2在化学通讯中的功能提供理论依据。利用RT-qPCR检测了SlitCSP2在蛹及雌、雄成虫各组织的基因表达水平;Westernblot方法检测了SlitCSP2在幼虫期和雄成虫头部、触角和足部的蛋白表达水平;利用免疫组织化学技术对Slit CSP2在幼虫各龄期的分布情况进行定位分析;利用荧光竞争结合试验检测Slit CSP2与13种非挥发性化合物的结合特性。RT-qPCR结果表明,SlitCSP2在3龄幼虫表达量最高,3～6龄表达量逐渐降低;成虫阶段雄成虫足部及胸部和雌成虫头部及翅的表达量最高,且雄成虫头部、足部和胸部的表达量显著高于雌成虫。Western blot结果显示,SlitCSP2在3龄幼虫头部（去触角）的蛋白表达水平较高。免疫组化结果表明,Slit CSP2在幼虫阶段主要分布于头部、口器和胸足;在雄成虫中,Slit CSP2主要分布于足部前跗节和足部外骨骼内侧。荧光竞争结合试验表明,Slit CSP2与单宁（Ki=1.77）、花青素（Ki=2...