【摘要】 目的 了解乙型肝炎病毒（HBV）X基因在HBV相關肝病肝移植受體術后外周單個核細胞（PBMC）和骨髓CD34+細胞內的整合情況及其對乙肝疫苗接種的影響。 方法 采集1999年6月-2005年11月25例HBV相關肝病肝移植受體的外周靜脈血及其中23例的骨髓血，以密度梯度離心結合單克隆免疫磁珠分離法獲取外周血單個核細胞及骨髓CD34+細胞后，提取細胞DNA。因HBV X基因的整合頻率最高，設計HBV X基因的特異引物，進行HBV-Alu-PCR，終產物進行電泳并回收、連接載體、篩選擴增后測序，檢測有無X基因整合。 結果 經PCR后電泳及測序分析，25例HBV相關肝病肝移植受體術后的PBMC內未檢測出HBV X基因的整合，其中采集到骨髓標本的23例CD34+細胞中亦未檢測到HBV X基因的整合。 結論 肝移植術后受體體內HBV微生態的劇烈改變，使HBV整合的基本條件喪失，在此情況下，外周免疫細胞及骨髓造血干/祖細胞不是發生HBV整合的適宜場所，乙肝疫苗接種效果與HBV X基因整合關系不明確。【Abstract】 Objective To investigate whether hepatitis B virus HBV X gene integrates in peripheral blood mononuclear cell (PBMC) and bone marrow CD34+ cells from HBV related liver disease recipients after liver transplantation. Methods Between June 1999 and November 2005, PBMC were obtained from 25 HBV related liver disease recipients after liver transplantation and bone marrow CD34+ cells obtained from 23 cases among them. The cellular DNA was extracted by DNA isolation and purification kit following the manufacture’s instructions. Specific primers to HBV X gene and to human Alu repeats were used to amplify the virus integration through a 3-round hemi-nest PCR. The PCR final product was judged by 1.2% agarose electrophoresis, ligated to T vector, proliferated in E. coil 5α and sequenced. Results According to agarose electrophoresis and sequencing analysis, there were no HBV X gene integration in PBMC and bone marrow CD34+ cells from HBV related liver transplant recipients after surgery. Conclusions Because of the radical change of HBV microecological environment in HBV related liver transplant recipients after operation, the fundamental condition of HBV integration has been lost, which led PBMC and bone marrow CD34+ cells not suit to HBV X gene integrate to human genome. And the impact of HBV X gene integration on HBV vaccination is still undefined.
【摘要】 目的 研究2型糖尿病（type 2 diabetes mellitus，T2DM）合并非酒精性脂肪性肝病（non-alcoholic fatty liver disease， NAFLD）患者血漿脂肪細胞特異性脂肪酸結合蛋白（adipocyte-specific fatty acid-binding protein，A-FABP）的水平及其相關因素。 方法 2009年10月—2010年10月選取T2DM合并NAFLD組（A組）60例，未合并NAFLD組56例（B組）為研究對象。測定體質量指數（body mass index，BMI），檢測血脂、糖化血紅蛋白（hemoglobin A1c，HbA1c）等生化指標。放射免疫法測定空腹胰島素（fasting insulin，FINS），空腹C肽水平（fasting C-peptide，FCP），計算胰島素抵抗指數（homeostasis model of assessment-insulin resistance，HOMA-IR）、胰島素敏感指數（insulin sensitivity index，ISI），測定A-FABP、C反應蛋白（C-reaction protein，CRP）及腫瘤壞死因子-α（tumor necrosis factor-α，TNF-α）。 結果 與B組患者相比，A組患者其血漿A-FABP水平、BMI、腰圍、腰臀比、丙氨酸氨基轉移酶、門冬氨酸氨基轉移酶、CRP、TNF-α、FCP、FINS、總膽固醇、甘油三酯、Ln（HOMA-IR）升高，Ln（ISI）降低，差異有統計學意義（Plt;0.05）；兩組HbA1c差異無統計學意義（Pgt;0.05）。A-FABP水平變化與TNF-α、HOMA-IR、CRP呈正相關，與ISI呈負相關。 結論 T2DM伴NAFLD中，A-FABP升高與胰島素抵抗是并存的，且存在明顯相關關系，二者在疾病的發生發展中均可能具有重要的作用。【Abstract】 Objective To analyze the serum level of adipocyte-specific fatty acid-binding protein (A-FABP) in patients with type 2 diabetes mellitus (T2DM) complicated with non-alcoholic fatty liver disease (NAFLD), and its related factors. Methods From October 2009 to October 2010, 112 patients with T2DM were categorized into two groups: the group with NAFLD (group A) with 60 patients, and the group without NAFLD (group B) with 56 patients. Body mass index (BMI), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), and hemoglobin A1c (HbA1c) were detected. Radioimmunoassay was carried out to measure fasting insulin (FINS) and fasting C-peptide (FCP), and homeostasis model of assessment-insulin resistance (HOMA-IR) and insulin sensitivity index (ISI) were calculated. At the same time, A-FABP, C-reaction protein (CRP) and tumor necrosis factor-α (TNF-α) were also detected. Results Compared with patients in group B, plasma levels of A-FABP, BMI, waistline, waist to hip ratio, ALT, AST, TG, TC, HDL-C, LDL-C, CRP, FCP, FINS, and HOMA-IR for patients in group A were all higher, while ISI was lower; and the differences in the above-mentioned parameters were statistically significant (Plt;0.05). The levels of HbA1c in the two groups were not significantly different (Pgt;0.05). The change of A-FABP level was positively correlated with TNF-α, HOMA-IR and CRP, while it wasnegatively correlated with ISI. Conclusions In patients with T2DM with NAFLD, there is an obvious correlation between the coexisting A-FABP rise and insulin resistance. Both of them played critical roles in the onset and developing of the disease.
