背景:抗瓜氨酸化蛋白抗体是类风湿关节炎特异性、早期诊断标志物,与类风湿关节炎发生发展及骨破坏的加重密切相关,该类抗体阳性患者的病情比阴性患者更严重而且更难治疗。胶原诱导型关节炎和佐剂诱导型关节炎等是典型的2种类风湿关节炎...背景:抗瓜氨酸化蛋白抗体是类风湿关节炎特异性、早期诊断标志物,与类风湿关节炎发生发展及骨破坏的加重密切相关,该类抗体阳性患者的病情比阴性患者更严重而且更难治疗。胶原诱导型关节炎和佐剂诱导型关节炎等是典型的2种类风湿关节炎动物模型,但都不表达或不能稳定高水平表达抗瓜氨酸化蛋白抗体,从而严重影响了抗瓜氨酸化蛋白抗体阳性类风湿关节炎发病机制和治疗药物研发的深入研究。目的:通过查阅国内外相关文献,综合分析抗瓜氨酸化蛋白抗体高表达的类风湿关节炎动物模型的研究现状,指出存在问题并提出建议。方法:以“Rheumatoid Arthritis,ACPA”或“Rheumatoid Arthritis,anti-CCP”为英文检索词,以“类风湿关节炎、抗瓜氨酸化蛋白抗体”为中文检索词,通过检索PubMed、Web of Science、中国知网、万方及维普数据库中2021年8月前发表的相关文献,最终纳入文献61篇进行综述分析。结果与结论:①抗瓜氨酸化蛋白抗体高表达的类风湿关节炎动物模型建立的主要方法有抗瓜氨酸化蛋白抗体单克隆抗体诱导、瓜氨酸化蛋白诱导和牙龈卟啉单胞菌诱导等,不同的造模方法各有特点。②瓜氨酸化蛋白诱导模型报道文献最多、建模后动物关节炎特征最明显,且该类蛋白为类风湿关节炎发病的靶抗原之一,能很好地模拟人体病理变化,当属建模首选模型。③氨酸化蛋白诱导和抗瓜氨酸化蛋白抗体单克隆抗体诱导方式类似,但均具有造模剂制备繁琐和难以获得的缺点。④牙龈卟啉单胞菌诱导模型经济成本较低、造模材料相对简单易得,但缺陷是建模后易合并牙周炎等并发症。⑤因此,未来研究寻找合适的抗瓜氨酸化蛋白抗体单克隆抗体、瓜氨酸化蛋白或短肽作为免疫诱导剂,并选择遗传背景和免疫学因素与人类风湿关节炎发病适合的动物,探寻牙龈卟啉单胞菌展开更多
Objective： To study the expression level of TRF1 （telomeric repeat binding factor 1） protein in human acute leukemia and relationship between expression level of TRF1 protein and telomerase, Methods： A quantitativ...Objective： To study the expression level of TRF1 （telomeric repeat binding factor 1） protein in human acute leukemia and relationship between expression level of TRF1 protein and telomerase, Methods： A quantitative Western±Blot technique was developed using anti±TRF1^33±277 monoclonal antibody and GST±TRFI purity protein as a standard to further determine the expression level of TRF1 protein in total proteins extracted from clinical specimens. Results： Bone marrow tissues of 20 acute leukemia patients were studied, 11 healthy donors＇ bone marrows were taken as a control. The expression level of TRF1 protein was significantly higher （P〈0.01） in normal bone marrow （（2.2174±0.462） μg/μl） than that of acute leukemia patients （（0.7544±0.343） μg/μl）, But there was no remarkable difference between ALL and ANLL patients （（0.6184±0.285） μg/μl vs （0.8454±0.359） μg/μl, P〉0.05）. After chemotherapy, TRFI expression level of patients with complete remission elevated （（0.7724±0.307）/μg/μl vs （1.6834±0,344）μg/μl, P〈0.01 ）, but lower than that of normal （（2.2174±0.462）/μg/μl, P〈0.01）. There was no significantly difference after chemotherapy （（0.7264±0.411） μg/μl vs （0.895±0.339） μg/μl,p〉0.05）. TRF1 expression level of patients with complete remission is higher than that of patients without complete remission （（1,683±0.344）μg/μl vs （0.895±0.339）μg/μl P〈0.01）. All samples were determined for telomerase activity. It was confirmed that the activity of telomerase in normal bone marrow was lower than that of acute leukemia patients （（0.125±0.078） μg/μl vs （0.765±0.284）μg/μl, P〈0.01）. There was no significant difference of expression level ofTRF I protein between ALL and ANLL patients （（0.897±0.290） μg/μl vs （0.677±0.268） μg/μl, P〉0.05）. After chemotherapy, telomerase activity of patients with complete remission decreased （（0.393±0.125） μg/μl）, but was展开更多
Tomato yellow leaf curl virus（TYLCV）is a species of the family Geminiviridae,causing serious yield losses in tomato production.The coat protein（CP）gene of TYLCV isolate SH2 was expressed in Escherichia coli BL21（...Tomato yellow leaf curl virus（TYLCV）is a species of the family Geminiviridae,causing serious yield losses in tomato production.The coat protein（CP）gene of TYLCV isolate SH2 was expressed in Escherichia coli BL21（DE3）using pET-32a as the expression vector.The recombinant protein was purified through Ni＋-NTA affinity column and used to immunize BALB/c mice.Three hybridoma cell lines（2B2,2E3 and 3E10）secreting monoclonal antibodies（MAbs）against TYLCV CP were obtained by fusing mouse myeloma cells（Sp 2/0）with spleen cells from the immunized BALB/c mouse.The titers of ascitic fluids of three MAbs ranged from 10-6 to 10-7 in indirect-ELISA.Isotypes and subclasses of all the MAbs belonged to IgG1,κ light chain.Triple antibody sandwich enzyme-linked immunosorbent assay（TAS-ELISA）showed that the MAb 3E10 could react with five begomoviruses infecting tomato,while the other two（2B2 and 2E3）mainly reacted with TYLCV.TAS-ELISA was set up using the MAb 3E10,and the established method could successfully detect virus in plant sap at 1：2 560（w/v,g mL-1）.Detection of field samples showed that begomoviruses were common in tomato crops in Zhejiang Province,China.展开更多
文摘背景:抗瓜氨酸化蛋白抗体是类风湿关节炎特异性、早期诊断标志物,与类风湿关节炎发生发展及骨破坏的加重密切相关,该类抗体阳性患者的病情比阴性患者更严重而且更难治疗。胶原诱导型关节炎和佐剂诱导型关节炎等是典型的2种类风湿关节炎动物模型,但都不表达或不能稳定高水平表达抗瓜氨酸化蛋白抗体,从而严重影响了抗瓜氨酸化蛋白抗体阳性类风湿关节炎发病机制和治疗药物研发的深入研究。目的:通过查阅国内外相关文献,综合分析抗瓜氨酸化蛋白抗体高表达的类风湿关节炎动物模型的研究现状,指出存在问题并提出建议。方法:以“Rheumatoid Arthritis,ACPA”或“Rheumatoid Arthritis,anti-CCP”为英文检索词,以“类风湿关节炎、抗瓜氨酸化蛋白抗体”为中文检索词,通过检索PubMed、Web of Science、中国知网、万方及维普数据库中2021年8月前发表的相关文献,最终纳入文献61篇进行综述分析。结果与结论:①抗瓜氨酸化蛋白抗体高表达的类风湿关节炎动物模型建立的主要方法有抗瓜氨酸化蛋白抗体单克隆抗体诱导、瓜氨酸化蛋白诱导和牙龈卟啉单胞菌诱导等,不同的造模方法各有特点。②瓜氨酸化蛋白诱导模型报道文献最多、建模后动物关节炎特征最明显,且该类蛋白为类风湿关节炎发病的靶抗原之一,能很好地模拟人体病理变化,当属建模首选模型。③氨酸化蛋白诱导和抗瓜氨酸化蛋白抗体单克隆抗体诱导方式类似,但均具有造模剂制备繁琐和难以获得的缺点。④牙龈卟啉单胞菌诱导模型经济成本较低、造模材料相对简单易得,但缺陷是建模后易合并牙周炎等并发症。⑤因此,未来研究寻找合适的抗瓜氨酸化蛋白抗体单克隆抗体、瓜氨酸化蛋白或短肽作为免疫诱导剂,并选择遗传背景和免疫学因素与人类风湿关节炎发病适合的动物,探寻牙龈卟啉单胞菌
文摘Objective： To study the expression level of TRF1 （telomeric repeat binding factor 1） protein in human acute leukemia and relationship between expression level of TRF1 protein and telomerase, Methods： A quantitative Western±Blot technique was developed using anti±TRF1^33±277 monoclonal antibody and GST±TRFI purity protein as a standard to further determine the expression level of TRF1 protein in total proteins extracted from clinical specimens. Results： Bone marrow tissues of 20 acute leukemia patients were studied, 11 healthy donors＇ bone marrows were taken as a control. The expression level of TRF1 protein was significantly higher （P〈0.01） in normal bone marrow （（2.2174±0.462） μg/μl） than that of acute leukemia patients （（0.7544±0.343） μg/μl）, But there was no remarkable difference between ALL and ANLL patients （（0.6184±0.285） μg/μl vs （0.8454±0.359） μg/μl, P〉0.05）. After chemotherapy, TRFI expression level of patients with complete remission elevated （（0.7724±0.307）/μg/μl vs （1.6834±0,344）μg/μl, P〈0.01 ）, but lower than that of normal （（2.2174±0.462）/μg/μl, P〈0.01）. There was no significantly difference after chemotherapy （（0.7264±0.411） μg/μl vs （0.895±0.339） μg/μl,p〉0.05）. TRF1 expression level of patients with complete remission is higher than that of patients without complete remission （（1,683±0.344）μg/μl vs （0.895±0.339）μg/μl P〈0.01）. All samples were determined for telomerase activity. It was confirmed that the activity of telomerase in normal bone marrow was lower than that of acute leukemia patients （（0.125±0.078） μg/μl vs （0.765±0.284）μg/μl, P〈0.01）. There was no significant difference of expression level ofTRF I protein between ALL and ANLL patients （（0.897±0.290） μg/μl vs （0.677±0.268） μg/μl, P〉0.05）. After chemotherapy, telomerase activity of patients with complete remission decreased （（0.393±0.125） μg/μl）, but was
基金supported by the Special Fund for Agro-Scientific Research in the Public Interest from the Ministry of Agriculture,China(201003065)
文摘Tomato yellow leaf curl virus（TYLCV）is a species of the family Geminiviridae,causing serious yield losses in tomato production.The coat protein（CP）gene of TYLCV isolate SH2 was expressed in Escherichia coli BL21（DE3）using pET-32a as the expression vector.The recombinant protein was purified through Ni＋-NTA affinity column and used to immunize BALB/c mice.Three hybridoma cell lines（2B2,2E3 and 3E10）secreting monoclonal antibodies（MAbs）against TYLCV CP were obtained by fusing mouse myeloma cells（Sp 2/0）with spleen cells from the immunized BALB/c mouse.The titers of ascitic fluids of three MAbs ranged from 10-6 to 10-7 in indirect-ELISA.Isotypes and subclasses of all the MAbs belonged to IgG1,κ light chain.Triple antibody sandwich enzyme-linked immunosorbent assay（TAS-ELISA）showed that the MAb 3E10 could react with five begomoviruses infecting tomato,while the other two（2B2 and 2E3）mainly reacted with TYLCV.TAS-ELISA was set up using the MAb 3E10,and the established method could successfully detect virus in plant sap at 1：2 560（w/v,g mL-1）.Detection of field samples showed that begomoviruses were common in tomato crops in Zhejiang Province,China.
