Development of super-low-invasive liver-biopsy-method using molecular genetics analysis

1.应用分子遗传学方法评价肝损害我们用寡核苷酸芯片分析了121例非肿瘤病变患者的基因表达谱。我们最终获得了81例患者的样本，这些患者可以用Azan染色进行肝脏平均纤维指数(MFI)分析。比较丙型肝炎和乙型肝炎患者中与干扰素或免疫功能相关的基因。在丙型肝炎病毒阳性的肝癌患者癌组织中，GIP2或IFI27在表达上调的区域均可观察到，居首位。该基因序列被鉴定为乙型肝炎和丙型肝炎患者不同的表达模式，许多基因被归类为C型和其他类型。超细针法分析结果评价18G和20G的总核糖核酸含量分别为30.4±4.4μg和15.3±1.4μg。此外，用较细的22G针提取总RNA量：4.9±0.9μg、23g：3.4±0.5μg，并对肝切除标本和超细针吸标本进行了聚类分析。基因的最终选择根据我们的上述结果，丙型肝炎和乙型肝炎的基因表达谱不同，我们认为丙型和乙型肝炎可以用不同的方法进行分析。首先分析了肝脏平均纤维化程度(MFI)与基因表达谱的关系。我们利用超细针抽吸对74个样本进行了微阵列分析，最终确定了39个与丙型肝炎肝损伤相关的基因，其中MHC-II类分子清楚地反映了炎症状态。此外，还包括干细胞激活基因，如‘PIK3C2B、ARHGDIB。这些基因可能是肝炎患者的分子预后因素。

1. Evaluation of liver damage using molecular genetics methodWe analyzed gene expression profile of non-cancer lesion of 121 patients using oligo array. We finally obtained samples from 81 patients who could be analyzed with liver mean fibrous index (MFI) using Azan staining.2. Comparing patients in hepatitis C and hepatitis BsAmong patients with hepatitis C, genes related Interferon or immune function were observed. GIP2 or IFI27, which located at the top of the list, were observed in upregulated expression area in cancer area of HCV-positive HCC patients. This gene list was identified to be different expression pattern of the patients with Hepatitis B and C. Lots of genes were classified in type C and others.3. The evaluation of result using analysis with super-thin-needle methodWe obtained the amount of total RNA using 18G:30.4±4.4μg and 20G:15.3±1.4μg. We confirm that the cluster using the same patients was located at the same position. Furthermore, we obtained the amount of total RNA using more-thin needle of 22G:4.9±0.9μg、23G:3.4±0.5μg. The cluster analysis were identified at the same cluster using liver resection sample and the super-thin needle aspiration sample.4. Final selection of genesAccording to our results above, gene expression profile of samples from hepatitis C and B were different, we consider that hepatitis C and B would be analyzed in different methods. At first, we analyzed the relationship of liver mean fibrotic change intensitiy (MFI) and gene expression profile.5. We analyzed a microarray analysis using 74 samples using super-thin-needle aspiration, then finally identified 39 genes that would be related liver damage of hepatitis C. Among them, MHC-class-II molecules were clearly reflected the inflammation status. Furthermore, stem cell activating genes, such as' PIK3C2B、 ARHGDIB, were included. These gene might be molecular-prognostic-factor of hepatitis patients.