鸡毒支原体特异膜抗原的制备

采用 Ｓ Ｄ Ｓ聚丙烯酰胺凝胶电泳分离纯化鸡毒支原体膜蛋白。选择切取１１０ Ｋ Ｄ、９８ Ｋ Ｄ、６６～６２ Ｋ Ｄ 和５６ Ｋ Ｄ 特异蛋白条带，以 Ｐ Ｂ Ｓ（ｐ Ｈ７．４）浸泡过夜，浸出液置于透析袋封口，透析袋外散布 Ｐ Ｅ Ｇ（ Ｍ ＝ ６ ０００）浓缩膜抗原后获得特异膜抗原，为鸡毒支原体特异诊断方法的建立和单克隆抗体的制备奠定了基础。     

Antigen-nonspecific macrophage factors modulating the antibody response in vitro

Antibody response to an antigen involves the co-operation between three types of cells: macrophages, T cells and B cells. The cognate interactions between these cells play a fundamental role in the expression of a specific antibody response, but the last is modulated by antigen-nonspecific soluble factors produced either by macrophages or by T cells. Macrophages elaborate a spectrum of molecules modulating the function of lymphoid cells; among them are IL1 and prostaglandins of the E series, which are respectively enhancer and inhibitor of the antibody response in vitro. These molecules alter T cell and B cell activities through different mechanisms involving activation or inhibition of IL2 production, or alteration of cells surface antigens. However, the cellular events following the fixation of soluble factor on its receptors are not known.     

担子菌原生质体融合子的免疫学鉴定

TritonX一100提取香菇单核体L52~(-adc)“和金针菇单核体F_(v23)的菌体壁抗原,分别注射兔子制备抗血清。同样方法制备L52~(-adc)“和F_(v23)原生质体融合子F_(02),F_(03)的菌体壁抗原,琼脂糖双向扩散显示融合子F_(02)、F_(03)的菌体壁抗原能与所制备的亲本抗血清发生特异性沉淀反应。表明两个融合亲本的菌体壁抗原基因在融合子中得到表达。从而也进一步证实融合子确为两亲本的杂合子。     

提高鸡传染性支气管炎病毒血凝抗原滴度途径及抗原灭活

将７株传染性支气管炎病毒（ＩＢＶ）标准株（Ｍ４１、Ｈ１２０、Ｈｏｌｔｅ、Ｇｒａｙ、Ｃｏｎｎｅｃｔｉｃｕｔ、Ｉｏｗａ６０９和Ｔ株）和６株分离株（ＮＩＢＶ、ＧＩＢＶ、Ｍ、ＳＨ、Ｊ和Ｈ株）分别接种于鸡胚，收获尿囊液，经浓缩后，用魏氏梭菌培养液处理，制备血凝抗原。其中，Ｈ１２０株血凝滴度最高，Ｔ、Ｍ、Ｊ和Ｈ株无血凝性。应用含有不同滴度ＩＢＶ母源抗体的鸡胚增殖病毒制备抗原，效价与用ＳＰＦ鸡胚增殖病毒制备的抗原效价一致。尿囊液经反复冻融后再制备抗原会使血凝价降低。抗原分别用甲醛、高碘酸钠、硼氢化钾和ＳＤＳ灭活，其中甲醛灭活效果最理想。抗原对氯仿敏感，对乙醚稳定。适宜浓度的Ｎａ＋、Ｍｇ２＋可显著提高抗原的血凝性     

Synthesis and Identification of Artificial Antigen for Imidacloprid

This study reported that a hapten of imidacloprid, 1-[(6-Carboxylethylthio-3- pyridinyl)methyl)-N-nitro-imidazolidinimine was synthesized using technical material of imidacloprid to react with 3-mercaptopropionic acid in hot alkaline solution. The hapten was conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to form the immuno-antigens and the coating antigen, respectively. The antibody with high titer (2.56 × 10⁴) was obtained after immuning rabbits. The results showed that the antibody had a high affinity to imidacloprid tested by enzyme-linked immunosorbent assay (ELISA), which IC₅₀ and IC₂₀ were 20.7 and 1.1 μg L⁻¹; and the cross reactibilities to those compounds related with imidacloprid were less than 1.4%.     

磺胺对甲氧嘧啶全抗原的制备研究

采用戊二醛法 ,将半抗原磺胺对甲氧嘧啶 (SMD)与载体牛血清白蛋白 (BSA)偶联 ,制得磺胺对甲氧嘧啶全抗原 SMD-BSA。其紫外扫描光谱和红外光谱表明偶联成功。通过改变半抗原 SMD和偶联剂戊二醛的用量 ,制得多种结合比的全抗原 ,这为进一步制备抗 SMD抗体提供了良好的免疫原     

Effect of prostate-specific membrane antigen on phospho-ERK,growth and migration of prostate cancer LNCaP cells

AIM: To study the expression of prostate-specific membrane antigen (PSMA) on the level of phospho-ERK, cell growth and migration of prostate cancer LNCaP cells.METHODS: The method of silencing PSMA was established by lentivirus-mediated RNAi in our early experiment. The cells were divided into 3 groups.In experimental group, the expression of PSMA in LNCaP cells was stably blocked by lentivirus-mediated RNAi. In negative control group, the cells were transfected with lentivirus-mediated control RNAi (without any interference to PSMA).The normal LNCaP cells served as blank control. The cells in these 3 groups were cultured in both 2 environments: normal medium and medium with PD98059 (an inhibitor of ERK phosphorylation). The phospho-ERK was detected by Western blotting and immunocytochemistry. Furthermore, the growth and migration of the cells were evaluated by MTT and transwell assays,respectively.RESULTS: In normal medium, the expression of phospho-ERK was attenuated in experimental group (P<0.05) and the quantity of "positive" cells was less than those in other 2 groups (P<0.05). Furthermore, the growth curves of the cells showed that the growth ability in experimental group was significantly decreased (P<0.05, after 48 h) and the migration ability in experimental group was reduced (P<0.05). In the inhibitory medium, the cells in all 3 groups expressed phospho-ERK at a lower level. Moreover, the abilities of growth and migration in these 3 groups were poorly displayed. These inhibitory effects on phosphorylation of ERK were similar to the cells in experimental group cultured in normal medium.CONCLUSION: PSMA may play a role in up-regulation of phospho-ERK and it may take an advantage in growth and migration of prostate cancer LNCaP cells.     

云南地区部分种猪场种公猪疫病的调查与分析

生产中,种公猪多种疫病会通过水平或垂直传播给种母猪或仔猪,造成种猪群繁殖障碍和呼吸道疾病暴发.为了解云南地区规模种猪场公猪疫病的感染情况,于2014年对云南10个地区63个种猪场4个品种的101头公猪血液和78头公猪精液进行了相关血清学和抗原检测.血清学检测结果显示:公猪血液中含有猪支原体、猪布氏杆菌病、猪弓形体病、猪传染性胸膜肺炎的抗体,其比例分别为32.67%、6.93%、8.91%、28.71%;其中,单独感染某一疫病30头占29.7%,二重混合感染有21头20.8%,三重混合感染2头1.98%,未见四重混合感染.对公猪精液进行PRV、CSFV、PRRSV、PCV-2、PPV和JEV的PCR检测发现:PRRSV感染检出率为12.82%,CSFV+PRRSV2二重混合感染检出率3.85%,PRRSV+PCV-2+JEV三重混合感染检出率2.56%.通过血清学和病原学共检测的179头种公猪,75头单独或混合感染以上10种病原,感染率42.13%,阳性猪场52家,占82.53%.     

Analysis of the distribution and clonality of TCR Vβ T cells in cord blood

AIM:To investigate the distribution and clonality of TCR Vβ subfamily T cells in cord blood. METHODS:The CDR3 of TCR Vβ 24 subfamily genes were amplified in mononuclear cells from 13 cases of cord blood. To observe the usage of TCR Vβ repertoire, the PCR products were further labeled with fluorescent and analyzed by genescan technique for the CDR3 size, to evaluate clonality of the detectable TCR Vβ T cells. Peripheral bloods from 10 cases of normal individuals and T cell line Molt-4 and Jurkat served as controls. RESULTS:Only 38.78%±16.26% of 24 Vβ subfamily T cell were selectively expressed in cord blood, predominantly in Vβ 3, 5, 8, 9 and 13, whereas all 24 Vβ subfamilies could be detected in T cells from peripheral blood of normal individuals. Genescan analysis showed that all PCR products of TCR Vβ subfamilies from cord blood or normal individual peripheral blood displayed multi-peaks. CONCLUSION:Some TCR Vβ subfamily T cells were absent in cord blood. All TCR Vβ subfamily T cells in cord blood displayed polyclonality.