目录

为了表达牛环形泰勒虫（Theileria annulata）截短HSP70基因编码蛋白并研究该蛋白的结构与功能特性,本研究扩增牛环形泰勒虫HSP70目的基因并构建重组质粒pMD18-T-HSP70,选取其他种的同源HSP70蛋白序列构建系统进化树;利用生物信息学方法分析HSP70基因编码蛋白的氨基酸组成、基本理化性质、亲疏水性、跨膜区结构、信号肽、可能的磷酸化位点、亚细胞定位及蛋白的二级结构和三级结构;对重组蛋白HSP70进行蛋白互作网络分析;构建原核表达载体pET28a-HSP70,筛选诱导表达条件,镍柱纯化重组蛋白及检测反应原性。结果显示,牛环形泰勒虫HSP70蛋白序列与小泰勒虫的序列同源性较高,蛋白分子质量为42 ku,理论等电点（pI）为5.61,属于酸性亲水性蛋白,无跨膜区及信号肽;蛋白功能预测结果显示,HSP70包含32个可能的磷酸化位点,亚细胞定位分析显示该蛋白主要分布于细胞质。蛋白质二级结构中α-螺旋、β-转角、无规则卷曲、延伸链分别占39.18%、8.51%、30.41%和21.91%。蛋白互作网络构建结果显示,与HSP70相互作用的蛋白主要为HSP90家族成员,另外还有伴侣蛋白GrpE同系物,预示着HSP70可能在细胞内与HSP90形成复合体发挥作用。本试验成功构建原核表达载体,获得了大小约为48 ku的融合蛋白,以0.6 mmol/L IPTG于37 ℃诱导5 h,蛋白表达较好;点状印迹及Western blotting结果表明,表达产物可被自然感染的牛环形泰勒虫阳性血清识别,具有良好的反应原性。本试验结果为进一步探讨牛环形泰勒虫HSP70功能机制提供了理论依据。     

奶牛常见繁殖疾病诊断与治疗

中国乳制品消费水平位于全球前三,奶牛养殖业在国内具有很大的发展潜力。随着奶牛养殖业的发展和扩大,繁殖疾病成为影响奶牛场经济效益的因素之一。繁殖疾病的发生和很多因素有关,包括营养、卫生等方面,体内激素不平衡也会诱发繁殖疾病的发生。本文对引起奶牛繁殖疾病的主要发病原因、临床症状、病理变化、临床诊断及治疗方法进行归纳总结,以期为繁殖疾病的诊断治疗提供理论基础。     

土地违法行为的经济学分析

行为人当其预期可以获得一定的经济利益时，便实施违法行为；相反，当其觉得无利可图时，便放弃这种行为。为此，试从经济分析的角度探讨土地违法行为的“成本”、“效益”关系，希望从成本、效益等因素人手，寻求遏制土地违法行为的有效途径以及健全土地法制的对策。     

抗滑桩优化设计在滑坡治理中的应用研究

基于传统剩余推力法，提出了新的抗滑桩优化方法。以位于凉山彝族自治州某县城南的滑坡体为例，说明了主要治理方法。采用剩余推力法和新的抗滑桩优化方对该滑坡体天然状态工况进行了稳定分析。结果显示，在设计安全系数为1．15的情况下，该滑坡处于临滑状态，因此，须采取相应的加固措施，最后推荐了抗滑桩设计方案，已在凉山州某县城滑坡工程中成功实施。     

Study on the Change Rule of Egg Yolk Antibody in Laying Hens after First Immunization by Japanese Encephalitis Virus

The purpose of this experiment was to study the immunization rule of the egg yolk antibody affected by different vaccines,immunization dose and injection ways and further to discuss the optimal immunization procedures of the laying hens for the preparation of egg yolk antibody against swine Japanese encephalitis virus.180 brown laying hens without any vaccines were selected and divided into 18 groups randomly,each group of 10 hens.Groups 1,2 were the control groups,injected with the sterile saline;Groups 3 to 10 were injected with subcutaneous or intramuscular injection,and the vaccine was injected with 0.2,0.5,1.0 and 1.5 mL successively.Groups 11 to 18 were also adopted two kinds of injection,followed by the same dose of vaccine immunization.Six eggs of each experimental group were gathered before immune day and after 3,7,10,14,18,21 and 28 days,the egg yolk antibody was extracted and the titer was determined.As a result,the egg yolk antibody titers of groups 1 to 6,11 and 12 were all 0,and no significant immune response produced;The hens from 7 to 10 groups were injected with the inactivated vaccine.After 7 days,the average antibody titer reached the peak,and the duration of the antibody was 14 days.The hens from 13 to 18 groups were injected with the attenuated virus vaccine.After 14 days,the average antibody titer reached the highest value,and the duration of the antibody was 21 days.The egg yolk antibody titers were not significantly different in the two compared experiment groups with the same injection dose but with different injection ways (P>0.05).With the same injection way of each experiment group,and the difference was significant (P>0.05).Compared with some groups with the same injection and vaccine,the titer of yolk antibody was gradually increased with the increase of the immune dose,and the difference was significant (P<0.05).The results showed that,no matter intramuscular or subcutaneous injection,in order to produce a significant immune response to hens,the immune antigen dose was 1.0 mL inactivated vaccine or 0.5 mL attenuated vaccine at least.Compared with the attenuated and inactivated vaccine,inactivated vaccine stimulated the body to produce the antibody faster,but the maintenance time was shorter;The lower dose of attenuated vaccine could stimulate the body to produce antibodies,but the speed was slower,the maintenance time was longer.     

Effects of lncRNA-GTL2 Transgenosis on Gut Microbiota Composition in Mice

This research was aimed to study whether the lncRNA would have effects on the structure and constitution of the intestinal microbial colonies in mice.High throughout sequencing was used to sequence and analyze the intestinal microbial colonies in both transgenic and non-transgenic mice of three months old.We conducted comparison and t-test at the level of phylum and genus.The result showed that transferred into long non-coding RNA genes GTL2 (lncRNA-GTL2),the mouse intestinal microbial colony structure and composition had no significant differences in the overall,within the same gender,but there were individual differences.Therefore,this study didn't find that long non-coding RNA transfered had a significant impact on the intestinal microflorain the phylum and genus level.     

东北地区奶牛乳房炎表皮葡萄球菌和腐生葡萄球菌的分子流行病学及耐药性研究

为调查表皮葡萄球菌和腐生葡萄球菌在东北地区的流行病学情况和耐药性,本研究对来自东北地区3个大型奶牛场采集的330份奶样进行葡萄球菌的分离、鉴定及其耐药表型的检测,并采用脉冲场凝胶电泳(PFGE)进行分离株的亲缘性分析,对表皮葡萄球菌进行多位点序列分型(MLST),同时应用PCR扩增分离株中携带的相关耐药基因。研究结果表明,在330份奶样中共分离到表皮葡萄球菌32株(9.7%),腐生葡萄球菌34株(10.3%);PFGE分析共获得9种不同谱型的表皮葡萄球菌和11种不同谱型的腐生葡萄球菌。药敏试验结果显示,两种菌对青霉素(70%)、苯唑西林(60%)和林克霉素(55%)的耐药率较高,主要耐药基因为lnu(B)(40%)、erm(B)(30%)和mec A(25%)。本研究结果揭示了东北地区奶牛乳房炎病原菌表皮葡萄球菌和腐生葡萄球菌的耐药谱和流行情况,为临床合理用药及奶牛乳房炎的防控提供了实验依据。     

桃在低温贮藏过程中品质变化研究

为比较不同类型桃在低温条件下的贮藏特性,以水蜜桃‘美香’和蟠桃‘银河’为试材,研究了低温贮藏过程中果皮颜色、果实硬度、可溶性糖和有机酸含量等的变化。结果表明:低温贮藏期间桃果皮颜色的变化在不同品种间表现有较大差异,‘银河’在贮藏过程中果皮颜色略微加深;2个品种果实硬度总体下降比较明显;不同品种果实可溶性糖含量的变化比较复杂,其中果实可溶性糖总量波动较小,蔗糖含量在贮藏后期明显下降;有机酸总量方面,‘美香’和‘银河’总体都呈现前期下降,至贮藏后期小幅上升的变化趋势,其中柠檬酸的含量在贮藏后期出现明显下降。     

International Practice of and Reference to the Standard Treaty Wording of Construction

Construction and relative service is one of the important categories in international service trade set down by WTO. In order to strike a construction agreement, the developed countries and districts have designed a lot of prevailing standard treaty wording, in which FIDIC plays the most important part. We should take advantages of their experience to improve our standard treaty wording.     

A New Method for Parameterized Design of Metallic Doors and Windows

According to the graphics characteristics of metallic doors & windows design and based on the research of graphics dimension restriction problems. This paper provides a new method for parameterized design of metallic doors & windows in virtue of expression solution. In this method, the restrictions of graphic elements have been changed into the parameter expressions of sizes and positions of graphic elements. Through the calculation of such expressions, the parameter driven method has been reached. Thus the parameterization of graphics can be easier and quicker than before. It can be used to setup the open parameterized graphics libraries.