氟喹诺酮类药物残留免疫学检测方法的研究

以沙拉沙星(SALX)为半抗原,牛血清白蛋白(BSA)偶联后免疫兔,获得对多种氟喹酮类(FQNS)药物有特异性的广谱性抗体.以SALX与卵清白蛋白(OVA)的偶联物作包被原,建立并优化了间接竞争ELISA检测方法,对SALX最低检出限达19.319 μg/L,且对诺氟沙星、环丙沙星和恩诺沙星均有特异性识别,最低检测限分别是22.646、26.181和19.054 μg/L.结果表明,该抗体可用于多种氟喹诺酮类药物残留筛选检测.     

猪源牛病毒性腹泻病毒套式PCR检测方法的建立

根据GenBank中BVDV-1、BVDV-2、CSFV及新出现瘟病毒的基因序列,设计2对特异引物,建立了能鉴别诊断BVDV与CSFV的Nested-PCR检测方法;确定其方法的特异性、敏感性、稳定性。建立的Nested-PCR能从BVDV标准毒株、猪源BVDV毒株中扩增198bp特异性片段,对猪瘟病毒、猪繁殖与呼吸综合征病毒检测结果为阴性。结果表明,该方法具有较好特异性;其敏感性可检测到0.195fg RNA模板量;经重复性试验表明该方法具有良好稳定性。初步应用检测表明,猪BVDV阳性率较高,达36.0%;牛血清及其制品、猪瘟活疫苗BVDV污染严重。本试验建立的Nested-PCR具有敏感、特异和稳定等特点,可用于临床诊断和流行病学调查。     

补铁剂及多肽铁复合物的研究进展

缺铁性贫血作为贫血中最常见的一种,越来越受到人们的关注。随着营养与健康需求的变化和安全意识的 增强,人们致力于补铁效果好、吸收好、副作用小、安全性更高的口服补铁剂的研发。本文对补铁剂的发展和相关 研究进行综述,概述新型补铁剂的研究进展,全面阐述多肽铁复合物蛋白琥珀酸铁的原料来源、结构、机理、制 备、应用、消化吸收机制,对多肽铁复合物和蛋白琥珀酸铁的发展前景进行展望,以期对补铁剂的研究与发展提供 理论依据。     

荒漠草原“科尔沁型”华北驼绒藜旱作技术研究

“科尔沁型”华北驼绒藜是优良的旱生牧草,但其果实带毛、粘连,不适宜机械播种,而且种子发芽要求较高的温度、湿度,在荒漠草原地区直接播种,建立旱作人工草地难度很大。采用育苗,机械化植苗技术,在撂荒、退耕地建立旱作人工草地,具有成活率高、速度快,操作性强、成本低、简单易行的特点,可实现大规模机械化作业,效益显著,为荒漠草原地区生态建设探索出了一条新的有效途径,可在北方干旱、半干旱地区大力推广应用。     

产肠毒素大肠埃希茵主要黏附素的研究进展

产肠毒素大肠埃希茵的主要黏附素抗原有K88、K99、987P和F41，在发病学和免疫学上扮演着重要的作用。文章就其理化特性、生物学特性、分子生物学特性及主要检测方法进行综述。     

Prokaryotic Expression of Brucella BAB Gene and Establishment of iELISA

In this study,the genome of Brucella Rev.1 strain was used as a template to amplify the BAB gene sequence,clone it into the prokaryotic expression vector pET-30a(+),obtain the recombinant plasmid pET-30a-BAB,and perform prokaryotic expression on the recombinant plasmid.The indirect ELISA method was constructed by using the expression products detected by Western blotting.The results showed that the BAB gene was successfully cloned and expressed in this experiment,and the purified expression product was analyzed by SDS-PAGE.This study obtained a relatively pure recombinant BAB protein;Western blotting test showed that the expressed protein could react specifically with Brucella sheep positive serum and had good reactogenicity;Using the recombinant BAB protein as the coating antigen,an indirect ELISA method for detecting BAB antibodies was established and optimized.The best determined coating conditions were as follows BAB protein coating amount was 0.25 μg/mL,serum dilution was 1:400;blocking solution was 3% pig-derived gelatin;secondary antibody dilution was 1:6 000;color development time was 10 min.The established method was used to detect 40 clinical sheep serums,and the cut-off value was calculated to be 0.607.That was,when the serum tested had P/N ≥ 1.5 and D_{450 nm} ≥ 0.607,it was judged as positive,when D_{450 nm} ≤ 0.561,it was judged as negative,and when 0.607<D_{450 nm}<0.561,it was judged as a suspect value,and retest was required.Compared with the Huhong plate test and the test tube agglutination test,the positive coincidence rate was 100%,the negative coincidence rate was 71.88%,and the total coincidence rate was 77.5%.     

家蚕马氏管组织构造的研究

通过对家蚕 (Bombyxmori) 5龄第 5日幼虫马氏管外部形态和内部组织结构的观察 ,发现不同部位马氏管的结构差异很大 :膀胱呈囊状体开口于小肠两侧 ,肌肉层厚、细胞层较薄 ,内腔很大 ;下行管靠膀胱处圆形 ,较膨大 ,其余大部分外型呈扁平形且表面光滑 ,细胞层较薄 ,内缘有微绒毛状突起、管腔较大 ;上行管前部管体稍扁平 ,结构与下行管相似 ,中后部管体圆形、细胞层肥厚、内缘呈蜂窝状、管腔狭小 ;隐肾管的外列马氏管细胞层薄、管腔较大 ,而内列马氏管细胞层肥厚、管腔很小。马氏管结构的差异与各部位的功能有密切关系。     

Study on Proliferation and Apoptosis of EBL Cells Induced by P48 Protein of Mycoplasma bovis

The aim of this study was to investigate the effect of P48 protein on proliferation and apoptosis of embryonic bovine lung (EBL) cells.In this study,samples which co-incubated with P48 protein and EBL cells in different concentrations at different time points were collected,the proliferation rate of the cells was detected by MTT method,and the changes in the nuclear morphology of EBL cells were observed by DAPI staining method.Meanwhile,flow cytometry was used to detect the apoptosis rate of EBL cells induced by P48 protein.Real-time quantitative PCR was used to detect the changes in mRNA level of apoptotic marker genes,and Western blotting tested the Bax and Beclin-1 protein expressions.The results showed that under the condition of 72 h and 10 μg/mL of protein concentration treatment,P48 extremely significantly inhibited EBL cells proliferation (P<0.01),while 0.1 and 0.5 μg/mL protein concentration had no inhibitory effect (P>0.05).The nuclear morphology showed no significant change after protein induction for 12 h,but wrinkled and condensed at 24 h.The nucleus was fragmented,and a sprouted apoptotic body was appeared at 48 and 72 h.Apoptosis related genes expression showed no obvious increase at 2 and 12 h at mRNA level,but gradually increased at 24,48 and 72 h,and it showed a time-dependent manner.Accordingly,the expression of apoptosis marker proteins Bax and Beclin-1 significantly increased.Flow cytometry analysis showed that the apoptosis rate of EBL cells induced by P48 protein was 48.44%.In conclusion,P48 recombinant protein of Mycoplasmas bovis inhibited the proliferation of EBL cells and promoted their apoptosis,which provided reference for revealing the pathogenic mechanism of Mycoplasmas bovis.     

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。