目的 本研究通过已建立的稳定表达人血小板CD36的HEK293T细胞系,制备鼠源的抗人CD36单克隆抗体并进行特性鉴定。方法 利用已经建立的稳定表达人血小板CD36的HEK293T细胞系对CD36(-/-)C57小鼠进行免疫,取脾脏与小鼠骨髓瘤细胞融合,筛选出阳性克隆。纯化单克隆抗体后,利用Western blot检测抗体结合活性。利用小鼠IgG类/单抗亚型鉴定试剂鉴定单克隆抗体的抗体亚型。通过流式细胞检测和血小板抗原单克隆抗体特异性免疫固定检测(MAIPA)鉴定其诊断应用价值。结果 经筛选后得到一株杂交瘤细胞,抗体亚型为IgG2a,轻链为κ链,Western blot试验表明该单克隆抗体特异性识别人血小板CD36抗原。MAIPA结果显示,与商业化单克隆抗体FA6-152相比,该单克隆抗体灵敏度更高。结论 成功制备了一种鼠抗人CD36单克隆抗体,为临床鉴定CD36抗体,筛选CD36阴性献血员提供了新的工具,也为今后进一步研究CD36在血液免疫疾病中作用机制提供了实验基础。

Objective In this study, by the established HEK293T cell line with stable expression of human platelet CD36, murine anti-human CD36 monoclonal antibody was prepared and characterized. Methods The established HEK293T cell line with stable expression of human platelet CD36 was used to immunize CD36 (-/-) C57 mice, and the spleen was fused with mouse myeloma cells to screen for positive cloning.After the purification of monoclonal antibody, the antibody binding activity was detected by Western blot.Mouse rapid antibody isotyping reagent was used to identify the subtype of monoclonal antibody.Its diagnostic value was evaluated by flow cytometry and monoclonal antibody-specific immobilization of platelet antigen (MAIPA). Results After screening, a high-producing hybridoma cell was obtained, the subtype of monoclonal antibody was IgG2a and the light chain was κ chain. Western blot analysis showed that the monoclonal antibody could specifically recognize CD36 antigen of human platelet. MAIPA results showed that the monoclonal antibody was more sensitive than the commercial monoclonal antibody FA6-152. Conclusion It is concluded that a mouse anti-human CD36 monoclonal antibody with biological activity has been obtained, which provides a new tool for the clinical identification of CD36 antibody and the screening of CD36 negative blood donors, and also provides an experimental basis for further research on the mechanism of CD36 in blood immune diseases.

目的 研究不同制备方式聚桂醇用于75岁以上患者下肢静脉曲张（VVLE）的疗效及耐受性观察。方法 纳入90例老年下肢静脉曲张患者作为研究对象,随机抽取分为两组,各45例。观察组在X线透视引导下行聚桂醇泡沫注射治疗,对照组行聚桂醇原液治疗。比较两组临床疗效和并发症,随访记录远期1年恢复效果。结果 观察组出院时治疗效果显著优于对照组（P < 0.05）,观察组皮下瘀血、血栓性浅静脉炎、肌间静脉血栓、术肢麻木及色素沉着并发症发生率低于对照组,两组间差异无统计学意义（P > 0.05）。两组患者远期恢复效果差异均无统计学意义（P > 0.05）。结论 透视引导下注射聚桂醇泡沫硬化剂治疗老年下肢静脉曲张疗效显著,安全性更高。