Fas对镉激活PC12细胞线粒体凋亡通路的影响

旨在探究死亡受体Fas在镉致大鼠肾上腺嗜铬细胞瘤细胞（PC12）凋亡中的作用及其对线粒体通路的调控机制,用10 μmol·L^-1镉处理Fas基因沉默的PC12细胞株12 h,通过Western blot检测BH3相互作用域死亡激动剂（BID）、半胱氨酸蛋白酶-9（caspase-9）、半胱氨酸蛋白酶-3（caspase-3）、多聚二磷酸腺苷核糖聚合酶（PARP）的活化情况,Bcl-2相关X蛋白（Bax）、B细胞淋巴瘤/白血病-2基因（Bcl-2）、凋亡诱导因子（AIF）、核酸内切酶G（Endo G）的表达情况,以及细胞色素C（Cyt C）在细胞内的分布情况,免疫荧光染色检测AIF核转位。结果显示,镉极显著上调tBID/BID比值和Bax/Bcl-2比值,诱导Cyt C从线粒体释放到细胞质,激活caspase-9、caspase-3和PARP,增加AIF和Endo G蛋白表达水平（P<0.01）,并诱导AIF核转位;沉默Fas极显著抑制镉引起的tBID/BID比值和Bax/Bcl-2比值升高,Cyt C从线粒体释放到胞浆,caspase-3、PARP蛋白活化和AIF、Endo G蛋白表达水平极显著升高（P<0.01）,显著抑制镉激活的caspase-9（P<0.05）,并抑制AIF核转位。综上表明,Fas通过调控线粒体通路参与镉致PC12细胞凋亡。     

磷对铝胁迫紫花苜蓿幼苗根系生长和生理特征的影响

为了探究磷对铝胁迫下紫花苜蓿幼苗生长和生理特征的影响,分别用不含和含200 μmol·L^-1磷（P）、100 μmol·L^-1铝（Al）、200 μmol·L^-1 P+100 μmol·L^-1 Al的简易 ［Ca（NO₃）₂］营养液（pH=4.5）处理铝敏感紫花苜蓿品种‘Wl440’幼苗。结果表明,在铝处理中添加磷后,苜蓿幼苗根系和叶片中的铝含量分别比铝处理降低81.53%和61.47%,苜蓿幼苗的根长和根系活力显著提高,叶片电导率和丙二醛（MDA）含量显著下降;光合生理得到明显改善,与铝处理相比,磷添加处理幼苗叶片的叶绿素含量、蒸腾速率、气孔导度和光合速率明显提高,光系统Ⅱ和光系统I的电子传递速率增加;磷添加处理明显提高了铝胁迫苜蓿根系的草酸和苹果酸含量,体内有机酸螯合铝离子的能力增强,光合能力提高。因此,磷能够通过增加根系有机酸含量,改善铝胁迫苜蓿光合系统,从而缓解苜蓿铝毒害。     

宁夏地区荷斯坦牛成母牛淘汰情况及长寿性影响因素分析

试验通过收集出生、产犊和离群的记录,探究宁夏地区荷斯坦牛成母牛的淘汰情况,分析影响该地区奶牛长寿性的因素。对15 523头荷斯坦牛的生产寿命、在群寿命和利用胎次进行描述性统计,分析淘汰时的胎次、季节和泌乳阶段的分布情况;计算生产寿命、在群寿命和利用胎次之间的表型相关,利用固定模型分析场-出生年、出生季节、牧场规模、淘汰原因和头胎产犊月龄对长寿性的影响。结果显示,宁夏地区荷斯坦牛的平均利用胎次为2.36胎,平均生产寿命为736.59 d。荷斯坦牛成母牛在产后第1个月内的淘汰风险最高,随着泌乳阶段的延长,成母牛在每个胎次内的淘汰风险均逐渐降低;随着淘汰胎次的增加,奶牛淘汰更加集中发生在产后泌乳早期;生产寿命、在群寿命和利用胎次之间存在较高的表型相关（均>0.9）,场-出生年、出生季节、牧场规模、淘汰原因和头胎产犊月龄对长寿性有显著影响（P<0.05）,春季出生和22月龄头胎产犊的奶牛长寿性表现更好。本研究初步揭示了宁夏地区荷斯坦牛成母牛的淘汰规律和长寿性的影响因素,为宁夏地区牛群选育成母牛长寿性状奠定了基础。     

北京市规模化奶牛场运动场防雨现状及建议

运动场是保障奶牛生产力和动物福利的重要设施。2021年6—9月,频繁降雨导致北京地区多家奶牛场的运动场积水,为奶牛生产力、奶牛场生防安全、环境卫生、运动场舒适性和动物福利带来隐患。通过线上线下结合,调研了北京地区27 家规模化奶牛场的运动场防雨、排水及垫料现状。结果表明：调研奶牛场中,88.90%奶牛场配备了运动场,其中83.33%奶牛场通过设置地面倾角、设置挡雨棚、雨污分离槽及布设排水管道（沟渠）、雨后水泵排水等方式实现运动场防雨及排水。此外,针对运动场建设、运动场防雨、运动场舒适性展开讨论,以期改善运动场防雨现状,满足集约化奶牛场可持续发展需求和提升动物福利。     

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。     

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。