BACKGROUND & AIMS In diagnostics, serum hepatitis B virus (HBV)-RNA levels are valuable when the HBV-DNA load in circulation is effectively suppressed by nucleos(t)ide analogue (NUC) therapy. This study aimed to determine the intrahepatic viral replication activity reflected in serum HBV-RNA and whether HBV-RNA contributes to liver histological changes in patients treated with NUC. METHODS A cross-sectional set of serum and liver biopsy samples was obtained from patients treated with entecavir, who had undetectable levels of serum HBV-DNA. The correlations between serum HBV-RNA concentration and levels of peripheral and intrahepatic viral replicative forms, as well as histological scores, were analyzed. Quasispecies of serum HBV-RNA and intrahepatic viral replicative forms were examined by deep sequencing. HBV-RNA-positive hepatocytes were visualized by in situ hybridization. RESULTS Serum HBV-RNA was detected in 35 of 47 patients (74.47%, 2.33-4.80log10copies/ml). These levels correlated not only with the intrahepatic HBV-RNA level and the ratio of intrahepatic HBV-RNA to covalently closed circular DNA (cccDNA), but also with the histological scores for grading and staging. Regarding quasispecies, serum HBV-RNA was dynamic and more genetically homogenous to simultaneously sampled intrahepatic HBV-RNA than to the cccDNA pool. In situ histology revealed that HBV-RNA-positive hepatocytes were clustered in foci, sporadically distributed across the lobules, and co-localized with hepatitis B surface antigen. CONCLUSION Serum HBV-RNA levels reflect intrahepatic viral transcriptional activity and are associated with liver histopathology in patients receiving NUC therapy. Our study sheds light on the nature of HBV-RNA in the pathogenesis of chronic HBV infection and has implications for the management of chronic hepatitis B during NUC therapy. LAY SUMMARY Serum HBV-RNA levels are indicative of the intrahepatic transcriptional activity of covalently closed circular DNA and are associated with liver histological changes in patients with chronic B hepatitis who are receiving nucleos(t)ide analogue therapy.

中文翻译：

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恩替卡韦治疗患者血清 HBV RNA 水平与肝内病毒及组织学活性标志物的关系

背景和目的在诊断学中，当循环中的 HBV-DNA 负荷被核苷（酸）类似物（NUC）治疗有效抑制时，血清乙型肝炎病毒（HBV）-RNA 水平是有价值的。本研究旨在确定反映在血清 HBV-RNA 中的肝内病毒复制活性以及 HBV-RNA 是否有助于接受 NUC 治疗的患者的肝脏组织学变化。方法 一组横断面的血清和肝活检样本来自接受恩替卡韦治疗的患者，这些患者的血清 HBV-DNA 水平检测不到。分析了血清HBV-RNA浓度与外周和肝内病毒复制形式水平以及组织学评分之间的相关性。通过深度测序检查血清HBV-RNA和肝内病毒复制形式的准种。通过原位杂交观察HBV-RNA阳性肝细胞。结果 47 例患者中有 35 例检测到血清 HBV-RNA（74.47%，2.33-4.80log10copies/ml）。这些水平不仅与肝内 HBV-RNA 水平和肝内 HBV-RNA 与共价闭合环状 DNA (cccDNA) 的比率相关，而且与分级和分期的组织学评分相关。关于准种，血清 HBV-RNA 是动态的，并且与同时采样的肝内 HBV-RNA 相比，与 cccDNA 库相比在遗传上更同质。原位组织学显示 HBV-RNA 阳性肝细胞聚集在病灶中，零星分布在小叶中，并与乙型肝炎表面抗原共定位。结论 血清 HBV-RNA 水平反映肝内病毒转录活性，并与接受 NUC 治疗的患者的肝脏组织病理学有关。我们的研究揭示了 HBV-RNA 在慢性 HBV 感染发病机制中的性质，并对 NUC 治疗期间慢性乙型肝炎的管理产生了影响。LAY Summary 血清 HBV-RNA 水平表明共价闭合环状 DNA 的肝内转录活性，并与接受核苷（酸）类似物治疗的慢性乙型肝炎患者的肝脏组织学变化有关。