目的 探讨早老素在日本血吸虫生殖发育及产卵中的作用.方法 分别收集日本血吸虫尾蚴感染小鼠后16、18、24 d后虫体,首先通过荧光定量PCR分析早老素在上述3个阶段表达;从感染后28 d雌虫中分离卵巢,荧光定量PCR分析早老素在卵巢的表达量.体外培养28 d雌雄合抱的成虫,应用siRNA对早老素进行干扰,10 d后,观察早老素被敲降后对血吸虫产卵量,雌雄合抱及生殖系统的影响.结果 荧光定量PCR结果显示,早老素在24 d雌虫表达量明显上调,同时证实该基因在卵巢组织高表达.当早老素被敲降后,雌雄合抱率和雌虫的产卵量也有明显下降.激光共聚焦结果显示相对于对照组和Mock组,RNA干扰组的生殖细胞减少,生殖系统形态出现异常.结论 早老素在日本血吸虫生殖发育和产卵中起重要调控作用.

Objective To investigate the anti-schistosomiasis effect of Tsp2/Sj29 ku protein from Schistosoma japonicum. Methods The recombinant protein pet32a/SjTsp2/Sj29 ku induced and expressed by IPTG was puri-fied. Twenty nine female 6-week old mice were randomly divided into 3 groups ( Protein vaccine Group, Thioredox-in Group and Control Group). In Protein vaccine Group (n=10), each mouse was immunized subcutaneously with 50 μg SjTsp2/Sj29 ku plus equal volume Freund's complete adjuvant in 0 day. 12 and 24 days later, each mouse was re-immunized subcutaneously with 50 μg SjTsp2/Sj29 ku plus equal volume Freund's incomplete adjuvant. In Thioredoxin Group ( n=10 ) and Control Group ( n=9 ) , each mouse was immunized with thioredoxin and saline respectively. Ten days after last immunization, each mouse was challenged with 30±2 cercaria of Schistosoma ja-ponicum. On the 45th day post-infection, the mice were exeuted. The worm and egg reduction rates were calculat-ed. The sera was collected from orbital blood. The levels of IgG1, IgG2a, IgG2b, IgG3 and IgM antibodies in mouse sera were detected by enzyme-linked immunosorbentassay ( ELISA) . The level of IFN-γ and IL-4 was also detected by ELISA. Results Pet32a/SjTsp2/Sj29 ku was obtained as expected. The worm reduction rates and egg reduction rates in Protein vaccine Group were 23. 95% (P<0.05)and 55. 57% (P<0.05) respectively, com-pared with the Control Group. There was no statistical significance ( P >0.05 ) between Thioredoxin Group and Control Group in worm reduction rates and egg reduction rates. In Protein vaccine Group, IgG1, IgG2a and IgG2b were remarkably higher than that of Control Group, while IgG3 and IgM did not remarkably change. In comparison with the Control Group, the levels of IFN-γand IL-4 of the mice in Protein vaccine Group were obviously augmen-ted (P<0.01). Conclusion The mice, which have been immunized with Tsp2/Sj29 ku show a partial immune protection;the protein vaccine of SjTsp2/Sj29 ku could induce the mixed immune responses of Th1 and Th2 .

目的 构建以及表达鼠白介素6 (IL-6)受体融合蛋白.方法 采用基因重组技术将鼠IL-6受体融合基因克隆到杆状病毒质粒Bacmid上,重组杆状病毒质粒在脂质体的介导下转染昆虫细胞Sf9,转染后收集重组杆状病毒,采用空斑实验检测第3代重组杆状病毒的滴度,并通过Western blot来确定目的蛋白的表达.结果 重组杆状病毒质粒在DH10Bac大肠杆菌中包装成功,并转染Sf9细胞后得到重组杆状病毒,第3代重组杆状病毒的滴度为5×107 pfu/ml,Western blot表明目的蛋白在Sf9细胞中表达.结论 成功表达了鼠IL-6受体融合蛋白,为进一步研究IL-6在炎症性疾病中的作用奠定了实验基础.