Serum levels of microRNAs can specifically predict liver injury of chronic hepatitis B

AIM:To investigate whether circulating microRNAs (miRNAs) can serve as molecular markers to predict liver injury resulted from chronic hepatitis B (CHB). METHODS:The profiles of serum miRNA expression were first generated with serum samples collected from 10 patients with CHB and 10 healthy donors (Ctrls) by microarray analysis. The levels of several miRNAs were further quantitated by real-time reverse transcriptionpolymerase chain reaction with serum samples from another 24 CHB patients and 24 Ctrls. Serum samples of 20 patients with nonalcohlic steatohepatitis (NASH) were also included for comparison. The comparison in the levels of miRNAs between groups (CHB, NASH and Ctrl) was analyzed with Mann-Whitney Utest. The correlation between miRNAs and clinical pathoparameters was analyzed using Spearman correlation analysis or canonical correlation analysis. The receiver-operator characteristic (ROC) curves were also generated to determine the specificity and sensitivity of each individual miRNA in distinguishing patients with CHB from Ctrls. RESULTS:miRNA profile analysis showed that 34 miRNAs were differentially expressed between CHB and Ctrl subjects, in which 12 were up-regulated and 22 down-regulated in CHB subject (fold change > 2.0 and P < 0.01). The median levels of miR-122, -572, -575 and -638 were significantly higher (P < 1.00 × 10 -5 ) while miR-744 significantly lower (P < 1.00 × 10 -6 ) in CHB compared with the Ctrl. The levels of miR-122, -572 and -638 were also higher (P < 1.00 × 10 -3 ) while the level of miR-744 lower in CHB (P < 0.05) than in NASH, although the difference between them was not as significant as that between CHB and Ctrl. ROC curve analysis revealed that the levels of miR-122, -572, -575, -638 and -744 in serum were sensitive and specific enough to distinguish CHB, NASH and Ctrl. Multivariate analysis further showed that the levels of these miRNAs were correlated with the liver function parameters. Most significantly, it was the scatter plot of principal component with the levels of these miRNAs, but not the parameters of liver function, which clearly distinguished CHB, NASH and Ctrl subjects. CONCLUSION:Serum levels of miR-122, -572, -575, -638 and -744 are deregulated in patients with CHB or NASH. The levels of these miRNAs may serve as potential biomarkers for liver injury caused by CHB and NASH.

小分子差异蛋白在HBV相关性肝病患者血清中的SELDI-TOF-MS筛选

目的:探讨乙型肝炎病毒(HBV)相关性肝病患者血清中小分子差异蛋白在HBV感染后病情发展过程中的意义.方法:采用蛋白芯片及表面增强激光解析电离飞行时间质谱(surface-enhanced laserdesorption/ionization time-of-flight massspectrometry,SELDI-TOF-MS)技术对已用乙腈去除高丰度蛋白的正常对照(NC)、乙型肝炎病毒携带者(ASCs)、慢性乙型肝炎(CHB)、肝硬化(LC)及原发性肝细胞癌(HCC)患者术前血清进行检测,筛选各自的血清小分子差异表达蛋白,并分别建立诊断模型.在蛋白数据库expasy寻找相关差异蛋白信息,对差异蛋白峰的可能结构及功能进行评价.结果:与NC组比,ASCs组有63个蛋白质波峰的强度值存在统计学差异(P<0.05),其中29个上调,34个下调;CHB组有57个,其中21个上调,36个下调;LC组有68个,其中33个上调,35个下调;HCC组有74个,其中28个上调,46个下调;通过对比分析,发现在4个病例组表达均为上调的m/z为15889.8,蛋白峰强度值在NC<ASC<CHB<LC组,HCC组较CHB和LC组低;11742.2蛋白峰强度值在NC<ASC<CHB<LC组和NC<ASC<CHB<HCC组,在LC和HCC组最高,用此蛋白峰诊断HBV感染相关性LC的灵敏度和特异度分别为90%和86.67%,诊断HCC的灵敏度和特异度分别为93.33%和83.33%.结论:成功去除高丰度蛋白和应用SELDI-T O F-M S技术筛选H B V感染相关性肝病患者血清中小分子差异蛋白,m/z为8709.7、13759.8、14004.0、15361.89、16072.3、2746.8、3449.1、3941.06、4098.3、9445.5的10个蛋白峰可能与HBV感染有关;m/z为15889.8的蛋白峰可能成为H B V感染后进展为LC早期诊断的标志物,而m/z为11742.2的蛋白峰也许是HBV相关性LC或HCC的一个重要标志.

Quantitation of HBsAg predicts response to entecavir therapy in HBV genotype C patients

AIM:To analysis the factors that predict the response to entecavir therapy in chronic hepatitis patients with hepatitis B virus (HBV) genotype C. METHODS:Fifty patients [hepatitis B e antigen (HBeAg)-negative:HBeAg-positive = 26:24] with HBV genotype C, who received nave entecavir therapy for > 2 years, were analyzed. Patients who showed HBV DNA levels ≥ 3.0 log viral copies/mL after 2 years of entecavir therapy were designated as slow-responders, while those that showed < 3.0 log copies/mL were termed rapid- responders. Quantitative hepatitis B surface antigen (HBsAg) levels (qHBsAg) were determined by the Architect HBsAg QT immunoassay. Hepatitis B core-related antigen was detected by enzyme immunoassay. Pre-C and Core promoter mutations were determined using by polymerase chain reaction (PCR). Drug-resistance mutations were detected by the PCR-Invader method. RESULTS:At year 2, HBV DNA levels in all patients in the HBeAg-negative group were < 3.0 log copies/mL. In contrast, in the HBeAg-positive group, 41.7% were slow-responders, while 58.3% were rapid-responders. No entecavir-resistant mutants were detected in the slow-responders. When the pretreatment factors were compared between the slow-and rapid-responders; the median qHBsAg in the slow-responders was 4.57 log IU/mL, compared with 3.63 log IU/mL in the rapid-responders (P < 0.01). When the pretreatment factors predictive of HBV DNA-negative status at year 2 in all 50 patients were analyzed, HBeAg-negative status, low HBV DNA levels, and low qHBsAg levels were significant (P < 0.01). Multivariate analysis revealed that the low qHBsAg level was the most significant predictive factor (P = 0.03). CONCLUSION:Quantitation of HBsAg could be a useful indicator to predict response to entecavir therapy.

2型糖尿病在慢性乙型肝炎进展到肝硬化中的作用

目的:探讨2型糖尿病(type2diabetes mellitus,T2DM)在慢性乙肝进展到肝硬化中的作用.方法:本研究纳入2007-01/2011-12在北京卫生部中日友好医院和四川省绵阳市科学城医院住院,符合诊断标准和纳入标准,不符合排除标准的慢性乙型肝炎和乙肝肝硬化患者.分析入选患者的人口统计学、临床、生化和代谢等相关资料,单因素分析比较两组人群在入选参数上的差异,二元非条件Logistic回归分析研究T2DM在慢性乙肝进展到肝硬化中的作用.结果:研究纳入符合条件的ChildA/B级的慢性乙肝患者91例和乙肝肝硬化患者201例,T2DM在两组人群中的发病率分别为11.0%(10/91)和33.8%(68/201),单因素分析提示二者存在明显统计学差异(P<0.001).二元非条件Logistic回归分析的结果发现,与慢性乙肝患者相比较,在控制了性别、年龄、体质量指数(体质量)、吸烟、饮酒和Child得分的情况下,T2DM是影响乙肝肝硬化独立的相关因素,OR值为4.434,95%可信区间为2.049-9.591(P<0.001).结论:T2DM在慢性乙型肝炎进展到肝硬化中发挥着重要的作用,可能促进慢性乙肝向肝硬化进展,但是仍需大规模前瞻性的研究进一步验证.

肝脏瞬时弹性成像对ALT低于2倍正常上限值的慢性乙型肝炎肝纤维化的诊断价值

目的：探讨肝脏瞬时弹性成像（ FS）在丙氨酸氨基转移酶（ ALT）﹤2倍正常上限值（ ULN）的慢性乙型肝炎（ CHB）患者肝纤维化中的应用价值。方法选择2014年3月至2015年3月诊治的ALT﹤2× ULN的CHB患者168例，运用FS进行肝脏硬度（ Stiffness）测量。所有患者均行肝穿刺活组织检查，以肝穿刺活组织检查病理结果为标准，Stiffness值与之对比，同时绘制受试者工作特征曲线，计算受试者工作特征曲线下面积，选定cut-off值。计量资料的组间比较采用Kruskal-Wallis H检验。结果随肝纤维化程度的提高，Stiffnes值逐渐增高，差异有统计学意义（ P﹤0.01或P﹤0.05），同一肝纤维化分期的CHB患者的Stiffnes随着肝组织炎症程度的加重而升高，在S0~S2肝纤维化分期中不同炎症组间比较差异有统计学意义（ P﹤0.05）。除S0期外，Stiffnes值与肝纤维化分期呈正相关（ r＝0.825，P﹤0.01）。FS检测肝硬化AUC为0.927，其中以15.1 kPa作为肝硬化诊断的界值，敏感度为88%，特异度为91%。结论 FS在ALT﹤2× ULN的CHB患者肝纤维化中具有较好的应用价值，尤其诊断肝硬化的准确性高，值得临床推广。