单细胞凝胶电泳检测慢性乙型肝炎肝癌患者的dna损伤

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1、暨南大学硕士学位论文单细胞凝胶电泳检测慢性乙型肝炎肝癌患者的损伤一，工八塑旦，丹参酮，，，，，℃，广暨南大学硕士学位论文单细胞凝胶电泳检测慢性乙型肝炎肝癌患者的损伤，，川乙暨南大学硕士学位论文单细胞凝胶电泳检测慢性乙型肝炎肝癌患者的损伤目录二’’”””‘”’’‘’二”’‘二’‘’’二刚舀……………………二”……“…对慢性······……乙型肝炎合并肝癌患者损伤的研究材料····························································……与方法结果································

2、·····································……讨论·····································································……对丹参酮抗慢·········……乙肝患者氧化损伤的研究材料与方法····························································……结··································································………果讨论·····

3、································································……影响技术因素探讨···················································……结论··············································································……参考文献········································································

4、……匕八附卧英文缩略词·····································································……攻读硕················································……士期间已发表的论文致谢···········································································……丫严暨南大学硕士学位论文单细胞凝胶电泳检测慢性乙型肝炎肝癌患者的损伤目舀，，又称单细胞凝胶电泳彗星实验，是一项在单细胞水平上

5、检测有核细胞断裂的新技术。，川单细胞水平是指该技术可以对单个细胞进行检测而不是对群体检测。与传统方法相比，技术更为简便、快一速、灵敏、价廉、重复性好、样品量少、无需放射性标记，具有极。目前高的实用价值已使用技术观察的细胞有人体淋巴细胞、、、、，，表皮细胞成纤维细胞翠丸细胞肿瘤细胞等动物有肝细胞四等。年，和最早报道直接定量检测单个细胞损伤将被检测细胞包埋于琼脂糖凝胶中，进行裂解，在弱碱性条件下使解螺旋，中和后以丫陡橙染色，用光度计测定绿色荧光表明为双链损伤与红色荧光表明为单链损伤的强度，两者比值定量可反映的损伤程度。但由于该技术条件要求十分严格，故未被广泛应用’。年

6、，和发明了微凝胶电泳技术将细胞包埋于琼脂糖凝胶中，铺于载玻片上，在去污剂和高盐溶液中溶解，然后在中性条件下短时间电泳，断片从细胞核向阳极移，，指向。行从而形成类似于彗星的尾巴阳极如果细胞中损伤严重，断片移行的距离也就较长。断片的移行可用嗅化乙咙，’。随，染色再用荧光光度计测定染色强度后对上述方法进行了改进，采用，了较严格的溶解条件使中性成为检测双链断裂的灵敏方法’。然而，中性条件下裂解和电泳只能检测双链断片，不能测定单链断片，而许多物质诱导形成的单链断片数量比双链断片高一倍，中性条件检测损伤明显不如碱性条件下敏感。因此，该技术有一定的局限性。，’年等提出碱性单细胞

7、凝胶电泳方法即目前广泛使用的单细胞凝胶电泳。在中性条件下，双链不能打开，只有暨南大学硕士学位论文单细胞凝胶电泳检测慢性乙型肝炎肝癌患者的损伤双链打，断片才能进入凝胶中。，开时在强碱性条件下双链被打开，释放出断裂的断片。在碱性条件还可促使降解，防止干扰，经染色后能，灵敏度大明显地显示断裂大提高。该技术目前。已广泛应用在测定损伤与修复中在哺乳类细胞中，的分子量很高，且具有严密的超螺旋结。，，构在理想的实验条件下正常细胞不受损伤外加电泳电场不会使在凝胶中移动。在中，细胞之所以从原位向电泳电场的阳极迁移，形成慧星状图像，其原理目前认为是，，’」在细胞核中是呈环状附着在核基

8、质上的包埋