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目的研究早熟凝集染色体(PCC)技术的方法学改进,为核和电离辐射事故时快速估算受照生物剂量提供良好的读片标本。方法取人的肘静脉血,以常规的PCC培养方法为标准,通过对培养中加入化学诱导剂花萼海绵体诱癌素A(CA)、有丝分裂阻断剂秋水仙素来研究收获PCC的形态和长度;通过对制片方法的改进来增强收获的PCC形态达到良好的状态;加入苦杏仁苷和细胞松驰素B(Cyt-B)2种细胞增殖促进剂在60Coγ射线大剂量照射的条件下,对促进周围血淋巴细胞增殖的作用。结果①以加入CA为35 nmol/L、秋水仙素30 nmol/L培养50 h后收获的PCC形态与结构最好,符合长度∶宽度比例要求的细胞率最高(47.4%);②制片中增加低渗液的用量至8 ml和延长时间至30 min并在37℃水浴下进行,同时改变固定液的比例为2∶1能增强收获良好的PCC标本;③照射0～20.0 Gy后,以质量浓度苦杏仁苷为150 mg/L和Cyt-B为0.2 mg/L共同培养收获的细胞增殖率最高,常规法培养最低。结论本研究的方法学改进可快速获得良好的PCC读片标本,为核和电离辐射事故时快速估算受照的生物剂量提供方便。
OBJECTIVE To study the methodological improvement of precocious aggregated chromosome (PCC) technology and to provide a good read sample for rapid estimation of irradiated biological dose for nuclear and ionizing radiation accidents. Methods The elbow venous blood of human was collected. The conventional PCC culture method was used to study the morphology and length of PCC harvested by adding calyx cavernosum-inducing A (CA) and mitotic arrestin colchicine to the culture. ; Through the improvement of the preparation method to enhance harvested PCC morphology to achieve a good state; adding amygdalin and cytochalasin B (Cyt-B) two kinds of cell proliferation enhancers in the 60Coγ-ray irradiation conditions, On the promotion of peripheral blood lymphocyte proliferation. Results ① The morphology and structure of PCC harvested after adding CA at 35 nmol / L and colchicine 30 nmol / L for 50 h were the best, and the highest cell percentage (47.4%) was obtained according to the requirement of length: width ratio; Increasing the amount of hypotonic fluid to 8 ml and prolonging the time to 30 min and performing the treatment in a water bath at 37 ° C while changing the ratio of the fixative to 2: 1 enhanced the harvested PCC specimens; ③ After irradiating 0-20.0 Gy with The highest proliferation rate was obtained when the concentration of amygdalin was 150 mg / L and Cyt-B 0.2 mg / L, and the lowest was obtained by the conventional method. Conclusions The methodological improvements in this study allow for quick and good PCC readings to facilitate the rapid assessment of irradiated biological doses for nuclear and ionizing radiation accidents.