Preparation of Atherosclerosis Model Using Guangxi Bama Mini-pig

The atherosclerosis (AS) model was prepared in Guangxi Bama mini-pig,and the atherosclerosis index (AI) was preliminarily identified in occurrence of AS to provide the basis for the preparation of related models.20 Guangxi Bama mini-pigs were randomly divided into control group and experimental group which were fed with high fat and cholesterol diet to prepare the AS model.Blood biochemical indexes were detected in the process of model preparation and the correlation between AI and the results of vascular slices was analyzed to initially draw up AI of AS.The results of vascular slices showed that the incidence rates of AS of Guangxi Bama mini-pigs in experimental group and control group were 20% and 0,respectively.The association analysis between the results of vascular slices and AI in the pathogenic Guangxi Bama mini-pigs preliminarily suggested that AI was above 3.8 and lasted for more than 3 months during the attack.     

响应曲面法优化草莓汁花色苷促溶技术的研究

为了加速溶出草莓汁中的花青毒,以哈尼草莓为试验材料,原花青素含量为检测指标,依据中心组合响应面设计,研究果胶酶处理对草莓中色素物质溶出的影响。结果表明:果胶酶处理的最佳工艺条件为果胶酶添加浓度为(质量比)1.70‰,45℃处理2.87h;方差分析结果表明模型高度显著(P=0.0002),失拟性不显著(P0.1)。该模型可进行草莓汁加工中花青素溶出的分析及预测。通过该技术处理,可以得到花青素含量高、颜色较好的草莓汁。     

Infections Investigation of ALV and REV in Guangxi Local Poultry Breeds and Sequence Analysis of gp85 Gene of ALV Isolates

In order to investigate the infection status of avian leukosis virus (ALV) and avian reticuloendotheliosis virus (REV) in the major local breeds of Qinzhou,Guangxi,totally 953 samples of egg white,cloaca swab and serum of Ma duck,Shitou goose,Tiejiao-Ma chicken,turkey and pigeon were collected from the representing flocks and detected by the commercial ELISA kits.ALV was isolated for the ALV p27 positive samples by culturing on DF-1 cells,and gp85 gene was sequenced.The results showed that the detections of ALV were negative in the samples except those of Tiejiao-Ma chicken,while REV antibody was found positive in Ma duck,Tiejiao-Ma chicken and turkey.The nucleotide sequences of gp85 gene of two isolates shared 94.5% identity with each other,and shared 86.9% to 94.9% with reference strains.The amino acid sequences of gp85 gene of two isolates shared 91.5% identity with each other,and shared 84.0% to 91.6% with reference strains.There were many variable sites in the hyper variable region hr1 and hr2,and the vr2 and vr3 variable regions were relatively conservative.Phylogenetic tree analysis showed that the two isolates shared the highest homology with SCAU11-XG strain.     

长期培肥我国典型黑土玉米氮肥效应的演变趋势

【目的】依托中国东北典型农田设置的公主岭黑土长期培肥试验,研究长期培肥条件下黑土地区玉米产量和氮肥效应的时间演变趋势,阐明氮肥利用率和土壤养分之间的相关性,为指导我国建立长期培肥模式和提高氮肥利用率提供依据。【方法】本文选择不同有机肥用量(M0、M30和M60)下不施肥处理(CK)、施氮肥(N)、施磷钾肥(PK)和平衡施肥(NPK),共计12个处理,研究长期培肥对玉米的氮肥农学效率、氮肥当季利用率和氮肥累积利用率的影响。【结果】有机肥料和化肥(NPK)对玉米均具有显著增产效果,有机肥化肥配施可降低玉米产量的变异系数(CV)和提高可持续性指数(SYI)从而保证玉米的持续稳产高产。平衡施肥条件下的氮肥农学效率、当季利用率和累积利用率随着施肥年限的延长呈上升趋势并且达到显著或极显著水平;常量有机肥区和高量有机肥区的土壤肥力得到显著提升,氮肥效应随时间演变趋势分别为变化不显著和显著降低趋势。氮肥利用率与土壤有机碳、土壤全氮、土壤全磷和土壤速效钾呈显著负相关。【结论】在相对较高的肥力条件下,黑土地区玉米氮肥效应呈显著下降或变化不显著的趋势将为高肥力条件下有机肥部分替代化肥提供科学依据。     

马铃薯微型原种播种机关键部件设计

农业产业和经济发展速度的加快及科学技术水平的提高，使机械设计和制造技术在农业产业中大规模的推广应用。马铃薯作为农业产业重要的组成部分，通过提高其种植技术和水平可以间接地发展其种植潜力。该文在马铃薯种植期间以微型原种播种机为例，通过对原种种植农艺的分析，对关键部件的设计方案展开研究。      

Analysis on Genetic Diversity and Phyletic Evolution of mtDNA Cytb Gene in Zhongdian Yak

In order to investigate the genetic diversity and the origin of evolutionary relationship of Zhongdian yak,we analyzed the complete sequence of 15 individuals Cytb gene,its sequence polymorphism was analyzed,and the phylogenetic tree was constructed.The results showed that the length of the nucleotide sequence were 1 140 bp,with nucleotide frequencies of 26.3%,31.8%,13.1% and 28.8% for T,A,G and C,respectively.Three haplotypes were identified of 15 individuals,with 3 polymorphic sites,including two conversions,one transversion,haplotype diversity was 0.2571 and nucleotide diversity was 0.00035.Phylogenetic analysis suggested that Zhongdian yak and Bos mutusc clustered firstly,then gathered with Bison bison,which indicated that there were high genetic similarity and closer genetic relationship,genetic similarity with other cattle genus was relatively low.Combining with the proof of molecular biology and paleontology,the result supported the point that Bos grunniens and Bos mutus were classified as an alone genus in Bovinae.     

传染性法氏囊病病毒主要结构蛋白VP2、VP1和VP2-VP1串联基因的原核表达及其初步应用

为获得传染性法氏囊病病毒(IBDV)特异性抗体检测用抗原VP2、VP1及VP2-VP1蛋白,分别设计引物扩增IBDV野毒株NN1172的VP2和VP1基因,并扩增VP2和VP1基因中抗原性和亲水性较好的重要区域,通过PCR扩增基因串联方法对截短的VP2和截短的VP1基因进行串联,首次获得VP2-VP1串联基因,并对VP2、VP1和VP2-VP1串联基因进行了原核表达和鉴定。结果成功构建了原核表达载体pET-VP2、pET-VP1和pET-VP2-VP1;诱导表达条件显示,3个重组质粒分别转入BL21菌株后经0.05 mmol/L IPTG诱导表达,分别得到分子量为69、114和63 kDa的VP2、VP1和VP2-VP1重组蛋白,且均以包涵体形式表达,3个重组蛋白分别于诱导后5、3和6 h时表达量最多。Western blot结果显示,表达的VP2、VP1和VP2-VP1蛋白与鸡抗IBDV阳性血清均具有良好的反应原性。以纯化的VP2、VP1和VP2-VP1蛋白作为包被抗原对传染性支气管炎病毒(IBV)、呼肠孤病毒(ReoV)、禽白血病病毒(ALV)和新城疫病毒(NDV)4种阳性血清检测均为阴性,表明所获得的纯化蛋白具有高度的特异性;对免疫了IBD灭活疫苗,IBD基因工程疫苗和IBD弱毒疫苗的商业鸡群进行抗体检测,结果均能显示疫苗免疫后机体抗体水平的变化趋势。本研究表明利用该原核表达系统所表达的3个蛋白均具有良好的免疫反应活性,为IBDV特异性抗体的检测和新型亚单位疫苗的研发奠定基础。     

巨型艾美耳球虫偏菱形样蛋白EmRP免疫原性分析

在前期研究中,从巨型艾美耳球虫(Eimeria maxima)cDNA文库中筛选到了免疫保护性抗原偏菱形样蛋白EmRP,本研究为揭示该抗原的免疫保护机理,进一步检测了其免疫原性。以E.maxima卵囊cDNA为模板,用RT-PCR技术扩增EmRP,并构建真核表达质粒pVAX1-EmRP,将其经腿部肌肉注射2周龄的雏鸡,3周龄加强免疫。分别于首次免疫后和加强免疫后1周,采集血清,用ELISA方法检测血清特异性IgG水平,用荧光定量PCR方法(qPCR)检测细胞因子水平,用流式细胞术检测CD4^+T淋巴细胞和CD8^+T淋巴细胞的比例,分析EmRP诱导的免疫反应。序列分析表明,EmRP含有偏菱形样蛋白超家族成员保守区域,为非跨膜蛋白,分子量约为28.4 kD。真核表达质粒pVAX1-EmRP免疫鸡后,与对照组相比,鸡体IFN-γ、IL-2、IL-10和IL-17等细胞因子转录水平显著提升;CD8^+和CD4^+ T细胞比例显著提高,而血清特异性IgG水平与对照组差异不显著。结果表明,EmRP诱导的免疫保护作用主要是由其诱导的细胞免疫反应实现的,可以作为研制E.maxima新型疫苗的候选抗原。