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应用寡核苷酸芯片技术分析了121例非癌性肝损害的基因表达谱。最后，我们从81例患者中获得了可以使用Azan染色进行肝脏平均纤维指数（MFI）分析的样本.对丙型肝炎和乙型肝炎患者进行比较，观察丙型肝炎患者干扰素和免疫功能相关基因。在HCV阳性HCC患者癌区的上调表达区域中观察到位于列表顶部的GIP 2或IFI 27。该基因列表被鉴定为B型肝炎和丙型肝炎患者的不同表达模式。大量的基因被归类为C型和其他类型.超薄针法分析结果评价18 G法测得总RNA量为30.4±4.4μg，20 G法测得总RNA量为15.3±1.4μg。我们确认使用相同患者的群集位于相同位置。用22 G细针测得总RNA量为4.9±0.9μg，23 G细针测得总RNA量为3.4±0.5μg。用肝切除标本和超薄针吸标本进行聚类分析，确定在同一聚类.根据以上结果，丙型肝炎和B型肝炎的基因表达谱存在差异，我们认为丙型肝炎和B型肝炎可以采用不同的方法进行分析。首先分析肝纤维化平均变化强度（MFI）与基因表达谱的关系.我们对74例丙型肝炎患者的肝组织进行了基因芯片分析，最终确定了39个与丙型肝炎肝损害相关的基因。其中MHC-II类分子能清楚地反映炎症状态。此外，还包括干细胞活化基因，如PIK 3C 2B、ARHGDIB。这些基因可能是肝炎患者的分子预后因子。

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.