水牛犊牛血浆外泌体的分离与鉴定

研究旨在从水牛犊牛血浆中分离外泌体,并对分离的外泌体进行分子生物学特征分析和鉴定。采用差速离心法分离3头水牛犊牛的血浆外泌体,并在透射电子显微镜下观察其形态,使用纳米粒度及Zeta电位仪对提取的外泌体进行粒径大小分析,应用Western blotting方法检测外泌体标记蛋白钙联蛋白（Calnexin）、肿瘤易感基因101（TSG101）、CD9、CD81在3头水牛犊牛血浆外泌体中的表达情况。结果显示,从水牛犊牛血浆中分离的外泌体形态多为圆形和椭圆形,直径在30~150 nm;Western blotting检测结果表明,在水牛血浆外泌体表面,特异性蛋白Calnexin、TSG101和CD81阳性表达,CD9不表达。本研究从形态学和分子生物学特征等方面证实研究获得的提取物为外泌体,由此说明,差速离心法可以成功分离提取水牛犊牛血浆中的外泌体,为水牛外泌体的后续研究提供了重要技术基础和参考。     

降水、氮沉降及放牧对草地生态系统凋落物分解的影响研究进展

草地生态系统对我国建设生态文明、提供生态系统服务、保障食品安全和维护边疆稳定有举足轻重的作用。全球气候变化和过度的人为干扰打破了草地生态系统原有的平衡及稳定性,造成草地退化、沙化和盐渍化现象严重。草地生态系统结构、功能与过程的稳定是保证草地生态系统稳定的基础。凋落物分解过程是草地物质循环的重要环节,对维持草地生态系统稳定性具有重要作用。草地生态系统凋落物分解对降水格局变化、大气氮沉降增加及放牧干扰的响应已有较多的研究。对目前国内外相关研究进行了梳理和总结,发现降水、氮沉降和放牧对草地生态系统凋落物分解的影响,因地理位置和气候变化以及草地类型的异质性而不同,也有针对同一地区开展的研究呈现不同结果的现象。目前对这三种干扰因素中的两因素或三因素交互作用对草地生态系统凋落物分解过程的影响研究较少,而研究气候变化及人为干扰对草地生态系统凋落物的影响对正确理解草地生态系统结构与功能,以及可持续利用与保护草地生态系统具有重要的意义。     

子长市果桑引种栽培管理技术

根据子长市近几年果桑引种试验示范的实践,总结提出要选栽抗旱耐寒性强的红果1号、红果2号、白玉王、黑珍珠等果桑品种,667㎡栽植180株为宜,中干树形养成,成龄果桑园宜春剪不宜夏伐,要重点做好桑椹菌核病防治工作。     

Cloning and Bioinformatic Analysis of L1 Gene of BPV from Guizhou Province

In order to study and analyze L1 gene of bovine papillomavirus（BPV）in Guizhou province,the L1 gene of BPV-GZ01 strain was amplified,cloned and sequenced using bioinformatic softwares and methods,and the secondary structure,tertiary structure,B-cell preponderant epitope,conserved domains analysis, transmembrane domain and signal peptide of L1 gene were predicted.The results showed that the length of L1 gene was 1 494 bp,encoding 497 amino acids.The L1 gene of BPV-GZ01 strain shared an amino acid identities of 98.6%,99.4%,98.4%,94.4% and 91.3%,and a nucleotide identities of 99.1%,99.8%,99.4%,87.6% and 82.8% with those of BPV2,BPV2-SW01,BPV2-AKS01,BPV13 and BPV1 strains,respectively.The results of phylogenetic tree analysis indicated that there was a close relationship between BPV-GZ01 and BPV2-SW01 strains.The prediction of secondary structure of L1 protein indicated that the random coil,extended strand and alphahelix took a higher percentage.The L1 protein was supposed contain 6 potential antigen epitopes.And no transmembrane domains and no signal peptide were found.The tertiary structure of L1 protein was curved spiral structure.These results provided a theoretical basis for immunologic diagnosis and further research of nucleic acid vaccine of BPV.     

Isolation and Identification of BPIV3 Genotype C

To identify the infection agents from Ningxia Hui Autonomous region, where feedlot cattle indicated bovine respiratory disease complex (BRDC), the M gene of the bovine parainfluenza virus type 3 was amplified by RT-PCR.The PCR product was ligated to pMD18-T vector and cloned to E.coli DH5α.The positive clones were sequenced and compared with the reference strains in GenBank by the molecular biology software.Sequence alignment results showed that a BPIV3 strain was isolated from the samples and named NX49, the M gene of NX49 included 1 056 nucleotides.Evolutionary analysis showed that the NX49 belonged to BPIV3 C genotype and shared 99.4% nucleotide identity with that of the SD0835 isolated in Shandong province.The characterization of the NX49 demonstrated that it was sensitive to temperature, acid and organic matter.The presence of Mg²⁺ showed no protection against the treatment at high temperature.The HA test suggested that the NX49 enables to agglutinate the guinea pig RBC at 4 ℃ and the titer was 1∶4.The study isolated a BPIV3 genotype C strain successfully, which facilitate the study of molecular evolution and epidemiology of BPIV3 in China.     

Cloning and Sequence Analysis of HA and NA Genes of One Strain of H6N6 Subtype Avian Influenza Virus Isolated from Guizhou Province

To investigate the epidemic situation of H6N6 subtype avian influenza virus (AIV) in Guizhou province,A/duck/Guizhou/013/2014 was isolated from Sansui duck in live poultry market of Guizhou in 2014,the hemagglutinin (HA) and neuraminidase (NA) genes of DK/GZ/14 were subjected to clone and sequence analysis.The results showed that HA gene had the highest nucleotide homologies (97.5%) with the duck-origin H6N6 subtype AIV isolated from Eastern China in 2009,and the strains of HA gene proteolytic cleavage sites was P-Q-I-E-T-R-G,which accordeol with the molecular characteristic of low pathogenic AIV (LPAIV).However,NA gene of A/duck/Guizhou/013/2014 had the highest nucleotide homologies (98.2%) with the duck-origin H6N6 subtype AIV isolated from Fujian in 2007.The phylogenetic tree showed that A/duck/Guizhou/013/2014 and Hunan strains located in the same branch,while three duck-origin H6N6 subtype AIV isolated from Guizhou in 2007 and A/duck/Guizhou/013/2014 located in the different branch for HA and NA genes in genetic evolution,which suggested that A/duck/Guizhou/013/2014 was far with the local H6N6 subtype.The results also clearly indicated that duck-origin H6N6 subtype AIV had genetic diversity in duck population in Guizhou.     

安徽乡村生态旅游区景观规划方法探讨研究

基于乡村生态旅游区景观规划设计与建设的必要性,对乡村生态旅游区景观的特点以及当前景观规划中存在的问题进行了分析。在回顾总结城市意向和旅游目的地意向研究成果的基础上,提出了乡村生态旅游区景观意向具有双重多维结构,其中,显性结构要素包括景观元素特性、空间结构和文化意蕴,隐形结构要素包括主题形象、旅游产品和功能设施。以景观意向为引领,探讨了由6个循环阶段构成的乡村生态旅游区景观规划方法,最后进行了实证研究。     

基于GIS的城市绿地系统景观生态评价

以哈尔滨市群力新区绿地系统为研究对象,在遥感和GIS技术支持下,从绿地类型的基本斑块特征、斑块类型等级、景观异质性及景观破碎化4个层面上选取14个指数,对群力新区绿地景观格局进行分析。结果表明:研究区绿地率低,各类型绿地斑块数分布失衡,面积差异较大,形状复杂程度各异。景观多样性较好,优势度明显,但缺失绿地类型,且分布不均匀,破碎化水平较高。针对研究区绿地景观格局中存在的问题,提出群力新区绿地生态规划建议,使研究区绿地系统具有良好的景观格局,更有效的发挥城市绿地的生态功能。     

景天属植物耐非生物胁迫研究进展

景天属植物作为园林绿化的重要植物材料已被广泛应用。了解其对非生物胁迫逆境的适应性,对在生态园林城市的建设和环境修复中景天品种的选择和利用具有重大意义。现综述了近年来我国在景天属植物耐高温、低温、干旱、涝、酸、盐、光和重金属等非生物胁迫研究方面的进展,对存在的问题及今后的发展方向提出了一些建议。     

香葱连作障碍的成因及其修复试验

选取不同连作年限(0、2、4、6、10、15 a)的葱田土壤,测定土壤的养分、微量元素和重金属含量,发现不同连作年限土壤中的铁、硼、镁等微量元素含量丰富,但是连作葱田土壤的容重、有机质含量和铵态氮含量有不同程度降低,硝态氮含量显著提高。用5种土壤改良剂对连作年限为15 a的葱田进行土壤淋施处理,试验结果表明,奥普尔冲施肥使土壤中的硝态氮含量降低11.57%,铵态氮含量提高50.27%,香葱产量提高2.66%,是最佳的土壤改良剂。