Expression and test of the neutralization Fab antibody against infectious bursal disease virus

为表达抗鸡传染性法氏囊病病毒(IbdV)抗体fAb并检测其中和活性,本研究将抗IbdV抗体的轻链(l)和重链片段(fd)基因分别克隆于P ET-27b(+)载体中,并转化于大肠杆菌rOSETTA(dE3)进行诱导表达。将l和fd片段包涵体蛋白变性后等量混合于复性液中,制备fAb并对其进行活性鉴定。结果显示l和fd蛋白相对分子质量大小分别为25 ku和28 ku。WESTErn blOT和ElISA检测结果表明,获得的抗体fAb大小约为50 ku,并且与VP2蛋白和不同病毒株均具有特异性结合能力。体外中和试验结果表明,获得的IbdV抗体fAb具有中和活性,可以有效阻断IbdV(b87株)对鸡胚成纤维细胞(df1)的感染。本实验获得的IbdV抗体fAb有望成为治疗Ibd的候选生物制剂,为研制治疗Ibd抗体制剂奠定了基础。To express the neutralizing Fab antibody against infectious bursal disease virus(IBDV),the gene of light chain(L)or heavy chain fragment(Fd) against IBDV was cloned into the prokaryotic expression plasmid,respectively,and then the recombinant L or Fd was expressed in E.coli Rosetta(DE3),respectively,and purified through sole denaturation and co-renaturation of inclusion body.Western blot results showed that the Fab was approximately 50 ku.ELISA results showed that the Fab exhibited binding ability and specify to VP2 for different IBDV strains.The results of neutralization test in vitro showed that the Fab exhibited neutralizing activity to IBDV-B87 strainin chicken embryo fibroblast(DF1) cells.The Fab antibody prepared in this study is expected to become a candidate drug for treatment of IBD,which laid the foundation for the treatment of IBD.黑龙江省应用技术研究与开发计划项目(GC13C104

Curcumin-induced Apoptosis Affects the Expression and Localization of hnRNP A2/B1 in Human Osteosarcoma MG-63 Cells

姜黄素(CurCuMIn)诱导处理的人成骨肉瘤Mg-63细胞,在光镜和电镜观察细胞凋亡的基础上,对HnrnP A2/b1在核基质中存在、分布及其与凋亡相关基因产物在Mg-63细胞中的共定位关系进行了研究.经姜黄素处理后,细胞出现染色质凝聚、细胞核固缩、凋亡小体等典型的细胞凋亡形态特征;双向凝胶电泳和质谱鉴定结果显示,HnrnP A2/b1存在于Mg-63细胞核基质蛋白组分中,在姜黄素处理后细胞核基质蛋白中表达下调.蛋白质印迹杂交结果,证实HnrnP A2/b1在姜黄素处理前后的Mg-63细胞核基质蛋白中的存在及其表达下调变化.免疫荧光显微镜观察显示,HnrnP A2/b1定位于Mg-63细胞核基质纤维上,经姜黄素处理后出现分布位置与表达水平变化.激光扫描共聚焦显微镜的观察结果显示,HnrnP A2/b1在Mg-63细胞凋亡过程中与bAX、bCl-2、fAS和P53等基因产物具有共定位关系,且其共定位区域发生了变化.研究结果证实了HnrnP A2/b1定位于核基质纤维上,是一种核基质蛋白,在姜黄素诱导人成骨肉瘤Mg-63凋亡过程中的表达与分布变化及其与凋亡相关基因的关系显然对Mg-63细胞凋亡具有重要影响,这为深入揭示肿瘤细胞凋亡的机制提供了重要科学依据和深入探索的新方向.The morphology of human osteosarcoma MG-63 cells under light microscopy and transmission election microscopy was altered when treated with curcumin.The distribution of heterogeneous nuclear ribonucleoprotein (hnRNP)A2 /B1 in the nuclear matrix and its colocalization with the apoptosis-related gene products were examined.The results demonstrated that curcumin induced typical apoptotic responses,such as chromatin agglutination,nuclei condensation,and apoptotic body formation.Twodimentional PAGE and mass spectrum analyses showed that hnRNP A2 /B1 was complexed with the nuclear matrix.Confirmed by Western blotting,curcumin treatment resulted in a down-regulation of hnRNP A2 /B1 expression.An altered distribution of hnRNP A2 /B1 with the nuclear matrix was observed in immunofluorescence assays.Laser confocal microscopy showed that hnRNP A2 /B1 colocalized with apoptosis-related molecules,including Bax,Bcl-2,Fas and p53,with different patterns before and after curcumin treatments.Our study showed that expression level and the distribution of hnRNP A2 /B1,a nuclear matrix protein,were altered by curcumin treatments and mought be involved in the process of apoptosis in MG-63 cells.国家自然科学基金项目(No.30871241)---

基于CTAC改性活性炭膜包用于水溶液中Cr(VI)的吸附去除及其再生性研究

使用多孔聚丙烯膜包覆阳离子表面活性剂十六烷基三甲基氯化铵(CTAC)改性活性炭,制成CTAC改性活性炭聚丙烯信封式膜包,用于吸附去除水中的Cr(VI),并考察了膜包材料的再生性。采用扫描电镜和红外光谱表征分别对CTAC改性活性炭进行了形貌和结构表征,考察了CTAC改性后国药粉末活性炭膜包在海水、湖水、自来水3种实际水样中的去除效果,去除率为88.5%~97.5%,相对标准偏差为0.25%~1.9%。采用NaOH溶液解吸,膜包在3次重复利用后,对Cr(VI)的平均去除率可达91.2%,在5次重复利用后,对Cr(VI)的平均去除率可达65.4%,仍有较好的去除效果。该CTAC改性结合膜包的方法有望为含铬废水治理提供普适性的吸附材料和方法

基于CTAC改性活性炭膜包用于水溶液中Cr(VI)的吸附去除及其再生性研究

使用多孔聚丙烯膜包覆阳离子表面活性剂十六烷基三甲基氯化铵(CTAC)改性活性炭,制成CTAC改性活性炭聚丙烯信封式膜包,用于吸附去除水中的Cr(VI),并考察了膜包材料的再生性。采用扫描电镜和红外光谱表征分别对CTAC改性活性炭进行了形貌和结构表征,考察了CTAC改性后国药粉末活性炭膜包在海水、湖水、自来水3种实际水样中的去除效果,去除率为88.5%~97.5%,相对标准偏差为0.25%~1.9%。采用NaOH溶液解吸,膜包在3次重复利用后,对Cr(VI)的平均去除率可达91.2%,在5次重复利用后,对Cr(VI)的平均去除率可达65.4%,仍有较好的去除效果。该CTAC改性结合膜包的方法有望为含铬废水治理提供普适性的吸附材料和方法

基于CTAC改性活性炭膜包用于水溶液中Cr(VI)的吸附去除及其再生性研究

使用多孔聚丙烯膜包覆阳离子表面活性剂十六烷基三甲基氯化铵(CTAC)改性活性炭,制成CTAC改性活性炭聚丙烯信封式膜包,用于吸附去除水中的Cr(VI),并考察了膜包材料的再生性。采用扫描电镜和红外光谱表征分别对CTAC改性活性炭进行了形貌和结构表征,考察了CTAC改性后国药粉末活性炭膜包在海水、湖水、自来水3种实际水样中的去除效果,去除率为88.5%~97.5%,相对标准偏差为0.25%~1.9%。采用NaOH溶液解吸,膜包在3次重复利用后,对Cr(VI)的平均去除率可达91.2%,在5次重复利用后,对Cr(VI)的平均去除率可达65.4%,仍有较好的去除效果。该CTAC改性结合膜包的方法有望为含铬废水治理提供普适性的吸附材料和方法