大豆GmGBP1基因的原核表达及重组蛋白纯化

将大豆中已克隆的一个新的SKIP基因(GmGBP1)构建到原核表达载体pGEX-6p-1上,导入大肠杆菌BL21(DE3)中,对其进行IPTG诱导.结果表明:在IPTG浓度为0.1 mmol·L-1,诱导时间为8h,诱导温度为37℃时,重组蛋白得到表达,分子量大约为95 kDa,SDS-PAGE电泳结果表明重组蛋白主要以包涵体形式存在,用8 mol·L-1尿素对其进行溶解后经GST柱纯化,得到较好的纯化效果.     

教学督导听课技巧与评课策略分析——督导听课心得

通过3位优秀教师的讲课风格诠释了授课艺术,通过自己督导期间听课实践总结出听课技巧;吸纳前人经验,提出评课策略及应该注意的问题。     

金融消费者保护问题的框架性思考

金融消费者是为生活消费需要而购买、使用金融商品或接受金融服务的人,具有追求盈利最大化、抗风险能力差以及弱势地位突出等特点。现阶段我国有关金融消费者权益保护方面的工作同发达国家相比还很不完善,存在缺乏专项立法、相关法律滞后、缺乏专业纠纷解决机制以及对金融消费者合法权益保护不到位等问题,急需国家相关部门出台相关政策文件改善和解决。     

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。     

光叶紫花苕研究状况的文献分析——基于CNKI数据库的文献计量

以中国知网的文献资料为数据源,利用CiteSpace文献计量工具,对1972-2019年光叶紫花苕研究领域的相关文献从关键词、单位和区域贡献、载文期刊、作者群体等方面进行计量分析,同时结合文献的阅读,系统的阐述了48年来光叶紫花苕的发展历程.结果表明,近年来我国学者对光叶紫花苕的研究越来越重视,出现频次较高的关键词有"绿肥""产量""多花黑麦草""草粉"等;《四川畜牧兽医》为该领域研究论文载文量最多刊物,其次是《草业科学》《畜牧兽医》,分别为10,8,8篇.凌新康、徐载春、匡崇义等几位作者发文较多,但大多数作者群体分布较为分散.当前国内关于光叶紫花苕的研究热点主要集中在饲用和肥田,同时光叶紫花苕对生态的修复作用也崭露头角,大有发展利用之势.     

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。     

猪戊型肝炎病毒ORF3蛋白影响HepG2细胞核黄素代谢的lncRNA-mRNA调控网络的鉴定

为初步鉴定并挖掘出猪戊型肝炎病毒(Hepatitis E virus,HEV)ORF3蛋白影响HepG2细胞核黄素代谢信号通路的lncRNA-mRNA调控网络,本试验通过构建腺病毒过表达载体,制备高滴度过表达腺病毒,介导猪 HEV-ORF3在HepG2细胞中实现过表达。Western blotting检测ORF3蛋白过表达成功后,运用lncRNA高通量组学测序,筛选出差异表达的lncRNA并进行靶向差异基因预测,对lncRNA靶向差异基因进行GO功能和KEGG通路富集分析,初步鉴定出与核黄素代谢信号通路相关的lncRNA-mRNA调控网络。Western blotting结果显示,在约为12 ku处出现目的条带,说明成功实现腺病毒介导猪HEV-ORF3在HepG2细胞中过表达。lncRNA高通量组学测序结果显示,共发现102个显著差异表达lncRNAs表达量上调,80个显著差异表达lncRNAs表达量下调。GO功能和KEGG通路富集分析显示,初步挖掘出lncRNA(MSTRG.13995.2)和lncRNA(MSTRG.1960.1)可能是与核黄素代谢信号通路相关的显著差异表达lncRNA,分别通过顺式调控其靶向基因APC-5和FLAD1来影响核黄素代谢信号通路。本试验初步鉴定出lncRNA(MSTRG.13995.2)-APC5和lncRNA(MSTRG.1960.1)-FLAD1可能是影响HepG2细胞核黄素代谢信号通路的lncRNA-mRNA调控网络。     

农业机械化推广模式创新分析

随着农业生产规模的扩大和机械化水平的提高,人们在进行农业生产的时候,已经普遍接受并采用机械化的生产模式,文章就农业机械化发展出现的问题进行了介绍,并提出了在我国进行农业机械化的推广的具体措施。     

Research Lactobacillus rhamnosusFermented Herbal and Bacillus subtilis Complex on Immune Performance,E.coli Infection of Broiler

This experiment was designed to study the complex of Lactobacillus rhamnosus fermented herbal and Bacillus subtilis on White Feather broiler immunity performance and impact of Escherichia coli infection.360 one-day-old broiler chickens were randomly divided into 3 groups with 4 replicates in each group and 30 chickens per replicate.The pretrial period lasted for 7 d,and the experiment lasted for 35 d.The chickens in the group Ⅰ(positive control group) and group Ⅱ(negative control group)were all only fed a basal diet,group Ⅲ was test group,by additive 1% fermented herbal preparations,groups Ⅱ and Ⅲ were intraperitoneal injection of 1 mL E.coli at 35 d,broiler mortality,immune organ index,cecalmicroflora content,immunoglobulin levels,IL-2 and IL-6 content were tested.The results showed that injection of E.coli caused massive death of chickens,group Ⅱtook up to 75.00%,it was significantly higher than groups Ⅰ and Ⅲ (P < 0.05),the mortality in group Ⅲ was significantly lower than group Ⅱ(P < 0.05),was only 23.33%.Injection of E.coli maked spleen index and thymus index of group Ⅱincreased significantly (P < 0.05),the spleen index and thymus index of groups Ⅰ and Ⅲ were no significant difference (P > 0.05).E.coli counts was significantly decreased after injectionin group Ⅲ (P < 0.05),but the number of intestinal Lactobacilli of group Ⅲ was significantly increased (P < 0.05),and inhibited the propagation of E.coli,the counts of E.coli in groups Ⅰ and Ⅲ were no significant difference (P > 0.05).At 42 d,the sIgA of the intestinal fluid in group Ⅲ were higher than that of groups Ⅰ and Ⅱ with 11.99% and 36.56%,respectively(P < 0.05).The serum IgG concentrations of group Ⅲ was higher than that of groupsⅠand Ⅱwith 14.68% and 28.15%,respectively(P < 0.05).At 42 d,the IL-2 content of group Ⅱ was the lowest,it was significantly lower than group Ⅲ(P < 0.05),the IL-6 of group Ⅲ was significantly lower than group Ⅱ(P < 0.05).     

Isolation and Identification of Pathogenic Bacteria Causing Dairy Cow Mastitis in Liaoning and Drug Sensibility Test of Pathogenic E.coli

In order to understand the main bacterial pathogen species causing dairy cow mastitis in Liaoning, as well as the characteristics of drug sensitivity of the pathogenic E.coli,the milk samples from 75 dairy cows with clinical manifestations for mastitis in certain large-scale dairy farm in Liaoning were collected.The bacteria in milk were cultured and isolated with biochemical methods and in vitro drug sensitivity tests were processed with the isolated E.coli strains.The results showed that the main bacterial pathogen for dairy cow mastitis were E.coli（separation rate 58.7%),S.aureus（64.0%）and S.agalactiae（54.7%),and multiple infection including double and triple infection were identified.The drug sensitivity tests on the isolated E.coli indicated that the E.coli isolates were highly resistant to sulfonamides（resistance rate>85%）and chloramphenicol（resistance rate>30%),and they were relatively low resistant to ampicillin（9.5%),ciprofloxacin（9.5%),ceftiofur（7.1%）and ofloxacin（4.8%).The results was able to provide reliable theoretical basis for prevention and control of dairy cow mastitis in Liaoning area.