摘要：目的:构建针对人膜联蛋白A2(ANXA2)的RNA干扰(RNAi)慢病毒表达载体,转染THP-1细胞,探讨ANXA2在抗磷脂抗体(APL)诱导单核细胞组织因子(TF)表达中的作用。方法:设计4条ANXA2特异性RNAi的寡核苷酸序列,与慢病毒载体pGCSIL-GFP连接,PCR及测序鉴定正确后,Western蛋白印迹法筛选出有效干扰序列。包装293T细胞获得重组慢病毒LV-RNAi-ANXA2,感染单核细胞株THP-1,观察细胞ANXA2 mRNA和蛋白表达下降程度。再用APL/β2GPI复合物刺激干扰后的THP-1细胞,观察TFmRNA表达及TF活性变化。结果:成功构建RNAi慢病毒载体并筛选出有效干扰片段;包装293T细胞后病毒滴度为3×1012TU/L;感染THP-1后细胞ANXA2 mRNA和蛋白均被沉默。APL/β2GPI复合物刺激干扰后的THP-1细胞,其TF表达下降至基础水平。结论:构建的慢病毒表达载体能显著抑制THP-1细胞ANXA2的表达,进而影响APL诱导的TF表达,证明ANXA2在APL诱导的单核细胞表达TF过程中具有重要作用。

摘要：抗磷脂抗体与反复流产有关。资料显示肝素可治疗反复流产,但肝素对抗磷脂抗体的影响知之甚少。为了解肝素治疗者妊娠中和分娩后血清抗磷脂抗体的变化及与当次和以后妊娠结局的关系设计该项前瞻性研究。 研究对象为1993年6月～1998年5月期间自发性流产≥3次的妇女36例,平均年龄34.1(21～42)岁,无导致流产的其它原因,狼疮抗凝和抗心磷脂抗体阳性。狼疮抗凝因子阳性的标准是:稀释凝血酶原时间(PT)高于正常对照水平,稀释活化部分凝血活酶时间(PTT)高于正常对照的2倍。抗心磷脂抗体阳性的标准是:ELISA方法测定IgG和IgM抗体≥1.0U/ml。就诊后一经诊断为妊娠即开始肝素治疗。

摘要：Objective: The aim of the study was to propose a set of tests to clarify the diagnosis of repeated implantation failure in patients undergoing in vitro fertilization (IVF). Study design: Fifty-nine patients with at least two unsuccessful IVF attempts were included in the study. Blood samples were evaluated for the presence of underlying thyroid abnormalities, antiphospholipid antibodies (aPL), increased levels of natural killer cells (NK), inherited thrombophilia and mouse embryo assay factor (MEAf). The same tests were performed on 20 normal fertile control patients. Results: Seventy-six percent of IVF patients showed at least one abnormal result. This incidence was higher with respect to that found among control patients (45% ). The prevalence of thyroid abnormalities, aPL and increased NK level was higher in IVF patients whereas no differences were observed in terms of prevalence of inherited thrombophilias and MEAf. Conclusions: A better understanding of reproductive failure mechanisms should allow an effective diagnostic flow chart and a focused therapeutic option for patients experiencing repeated IVF failure. With this objective in mind, our data provide two important results: thyroid abnormalities, aPL and increased NK levels are more prevalent in women experiencing IVF failure. No evidence was found for an association between inherited thrombophilia and MEAf and failure to achieve pregnancy after IVF.