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采用细针吸引肝细胞和新鲜组织块流式细胞光度法(FCM)对比分析大鼠正常肝脏、肝硬化、非典型增生和肝癌组织,以探讨其对肝脏病变程度估计及肝癌早期诊断的可行性及临床意义。材料与方法1.动物模型制备体重120g 左右的 Wistar 大鼠70只,分为两组:一组为60只饮用150ppm 的二乙基亚硝胺水溶液,另组10只饮用自来水,饲养1.5个月。2.材料收集将大鼠在全麻下剖腹,用7号注射用针头抽吸肝脏,后在同一部位取活组织块,均固定于70%冷酒精中,再处死动物,10%甲醛固定肝脏,依据病理改变分为:正常肝脏、肝硬化、非典型增生和肝癌四组。将细针吸引物和组织块分别制成单细胞悬液,后采用溴化乙啶一步插入法将细胞悬液中的 DNA 定量染色。使用 FACS420型流式细胞仪测定 DNA 含量,根据细胞核发出的荧光信号和光散射信号,经计算机分析得出细胞核相对 DNA 含量的分布频率。
Flow cytometry (FCM) was used to compare the normal liver, liver cirrhosis, atypical hyperplasia, and hepatocellular carcinoma tissues with fine needles to attract hepatocytes and fresh tissue masses in order to explore the feasibility of estimating liver lesions and early diagnosis of liver cancer. And clinical significance. Materials and Methods 1. Animal Model 70 Wistar rats weighing about 120 g were prepared and divided into two groups: one group consisting of 60 drinking 150 ppm diethylnitrosamine aqueous solution and the other group drinking 10 tap water for 1.5 months. . 2. Materials collection Rats underwent abdominal laparotomy under general anesthesia, and the liver was aspirated with a 7-gauge needle. After that, the biopsy pieces were taken from the same site and fixed in 70% cold alcohol. The animals were sacrificed and the liver was fixed with 10% formaldehyde. According to the pathological changes were divided into: normal liver, cirrhosis, atypical hyperplasia and liver cancer four groups. The fine needle aspiration and the tissue block were each made into a single cell suspension, and the DNA in the cell suspension was quantitatively stained by one-step insertion of ethidium bromide. The FACS420 flow cytometer was used to measure the DNA content. According to the fluorescence signal and light scattering signal emitted from the nucleus, the distribution frequency of the nuclear DNA relative to the DNA content was determined by computer analysis.