Serum HBV DNA plus RNA reflecting cccDNA level before and during NAs treatment in HBeAg positive CHB patients.

Background & Aims: Correlations between serum viral markers and intrahepatic cccDNA in patients undergoing long-term nucleos(t)ide analogues (NAs) treatment haven't been fully explored. In this study, we evaluate the correlation between intrahepatic cccDNA and other serum viral markers and intrahepatic HBV DNA in HBeAg positive chronic hepatitis B (CHB) patients during 60-month treatment with NAs. Methods: Fifty-four HBeAg positive CHB patients received long-term NAs treatment were included in this study. Serial serum samples were regularly collected and quantitatively analyzed for HBsAg, HBV DNA, HBV RNA and HBcrAg. Histological samples from liver biopsy at baseline and month 60 were analyzed for intrahepatic HBV DNA and cccDNA. Results: At baseline, serum HBV DNA plus RNA was positively associated with intrahepatic cccDNA in multivariate regression analysis (ß=0.205, P<0.001). In the correlation analysis between cccDNA and serum viral markers, HBV DNA plus RNA had the highest correlation coefficient (r=0.698, P<0.001), followed by serum HBV DNA (r=0.641, P<0.001), HBV RNA (r=0.590, P<0.001), and HBcrAg (r=0.564, P<0.001). At month 60, correlations between these serum viral markers and cccDNA were not observed (P>0.05). Multivariate regression analysis showed that only the decreased HBV DNA plus RNA was positively associated with cccDNA decline (ß=0.172, P =0.006). Changes of HBV DNA plus RNA (r=0.525, P=0.001) was better correlated with cccDNA decline as compared to HBV RNA (r=0.384, P=0.008), HBV DNA (r=0.431, P=0.003), and HBsAg (r=0.342, P=0.029). Conclusions: Serum HBV DNA plus RNA better correlated with intrahepatic cccDNA than other viral makers before and during NAs treatment in HBeAg positive CHB patients.

Catheter ablation for atrial tachycardias: How to interpret the unclear activation map?

BACKGROUND: Post-ablation atrial tachycardias (ATs) are characterized by low-voltage signals that challenge current mapping methods. In this study, we analyzed common mistakes during activation mapping and delineated a mapping strategy for correct interpretation of tachycardia mechanisms in patients with challenging underlying substrate. METHODS AND RESULTS: Thirty-one patients referred for AT ablation were selected for the study. Three types of incorrect activation patterns were identified, which were referred to as unrecognized block line (pseudo-macroreentry and pseudo-fig-8 reentry), incorrect activation timing window of interest (WOI) (chaotic activation), and mis-annotation of complex signals (multiple sites of "early meets late"). Pseudo-macroreentry and chaotic activation occur in focal or localized reentry AT with the error related to the WOI selection (four cases), incorrect annotation of local activation time (six cases), or a previous line of atrial block in (seven cases). Pseudo-fig-8 reentry (five cases) and multiple sites of "early meets late" (nine cases) occur in macroreentrant AT with blocked areas and low-voltage atrial substrate. All ATs were successfully eliminated at the origin site. CONCLUSIONS: We delineated a series of ATs in the setting of a disordered pattern of activation mapping encountered in patients after previous extensive ablation or atriotomy. The algorithm proposed rapidly corrects the activation map and identifies the mechanism of the AT.

Yin-Yang 1 and HBx protein activate HBV transcription by mediating the spatial interaction of cccDNA minichromosome with cellular chromosome 19p13.11.

HBV cccDNA stably exists in the nuclei of infected cells as an episomal munichromosome which is responsible for viral persistence and failure of current antiviral treatments. However, the regulatory mechanism of cccDNA transcription by viral and host cellular factors is not well understood. In this study, we investigated whether cccDNA could be recruited into a specific region of the nucleus via specific interaction with a cellular chromatin to regulate its transcription activity. To investigate this hypothesis, we used chromosome conformation capture (3C) technology to search for the potential interaction of cccDNA and cellular chromatin through rcccDNA transfection in hepatoma cells and found that cccDNA is specifically associated with human chromosome 19p13.11 region, which contains a highly active enhancer element. We also confirmed that cellular transcription factor Yin-Yang 1 (YY1) and viral protein HBx mediated the spatial regulation of HBV cccDNA transcription by 19p13.11 enhancer. Thus, These findings indicate that YY1 and HBx mediate the recruitment of HBV cccDNA minichromosomes to 19p13.11 region for transcription activation, and YY1 may present as a novel therapeutic target against HBV infection.

Long-term functional maintenance of primary human hepatocytes in vitro.

The maintenance of terminally differentiated cells, especially hepatocytes, in vitro has proven challenging. Here we demonstrated the long-term in vitro maintenance of primary human hepatocytes (PHHs) by modulating cell signaling pathways with a combination of five chemicals (5C). 5C-cultured PHHs showed global gene expression profiles and hepatocyte-specific functions resembling those of freshly isolated counterparts. Furthermore, these cells efficiently recapitulated the entire course of hepatitis B virus (HBV) infection over 4 weeks with the production of infectious viral particles and formation of HBV covalently closed circular DNA. Our study demonstrates that, with a chemical approach, functional maintenance of PHHs supports long-term HBV infection in vitro, providing an efficient platform for investigating HBV cell biology and antiviral drug screening.

Catheter Ablation of Idiopathic Left Posterior Fascicular Ventricular Tachycardia: Predicting the Site of Origin via Mapping and Electrocardiography.

BACKGROUND: We report the 12-lead ECG morphology of left posterior fascicular ventricular tachycardia (LPF-VT) and the relationship between His-ventricular (HV) interval and site of origin in LPF-VT. METHODS AND RESULTS: We studied 41 patients who underwent successful catheter ablation of LPF-VT with HV interval >0 ms (n=8; proximal-LPF group), HV interval 0 to -15 ms (n=15; middle-LPF group), and HV interval <-15 ms (n=18; distal-LPF group). The earliest mapped presystolic potential (PP)-QRS interval was 34.1±4.2, 24.5±3.2, and 19.4±2.8 ms in proximal-, middle-, and distal-LPF groups. The earliest PP ratio (PP-QRS interval during VT/HV interval during sinus rhythm) was 0.59±0.05, 0.45±0.07, and 0.31±0.05 in the proximal-, middle-, and distal-LPF groups. There were statistically significant differences between the 3 groups in earliest PP ratio, and there was close correlation between the HV interval during LPF-VT and earliest PP ratio. The QRS duration in the proximal-LPF group (114±6 ms) was significantly narrower compared with the middle-LPF group (128±5 ms) and distal-LPF group (140±6 ms). In leads I and V6, the ratio of R/S tended to be greater in the proximal-LPF group compared with the other 2 groups. QRS duration, the ratio of R/S in leads V6, and lead I could predict a proximal or distal origin site of LPF-VT with high sensitivity and specificity. CONCLUSION: The HV interval and 12-lead ECG morphology of LPF-VT may help predict the successful site of origin and prove useful in guiding an effective ablation strategy.

STABLE-SR (Electrophysiological Substrate Ablation in the Left Atrium During Sinus Rhythm) for the Treatment of Nonparoxysmal Atrial Fibrillation: A Prospective, Multicenter Randomized Clinical Trial.

BACKGROUND: Circumferential pulmonary vein isolation (CPVI) alone or combined with adjuvant substrate modifications is unsatisfactory for atrial fibrillation (AF) control in nonparoxysmal AF patients. Ablation targeting the fibrotic areas after CPVI (STABLE-SR [Electrophysiological Substrate Ablation in the Left Atrium During Sinus Rhythm]) is a newly evolved substrate modification strategy. METHODS AND RESULTS: In this multicenter, randomized clinical trial, 229 symptomatic nonparoxysmal AF patients were 1:1 randomized to STABLE-SR group (n=114) or conventional STEPWISE group (n=115). In the STABLE-SR group, after CPVI, cavotricuspid isthmus ablation and cardioversion to sinus rhythm, left atrial high-density mapping was performed. Areas with low-voltage and complex electrogram were further homogenized and eliminated, respectively. Dechanneling would be done if necessary. In the STEPWISE group, additional linear lesions and defragmentation were performed.The primary end point was freedom from documented atrial tachyarrhythmias for ≥30 s after a single ablation procedure without antiarrhythmic medications at 18 months. At 18 months, 74.0% of the patients in the STABLE-SR group and 71.5% in the STEPWISE group (hazard ratio, 0.78; 95% confidence interval, 0.47-1.29; P=0.325) achieved success according to intention-to-treat analysis. However, less procedure time (186.8±52.7 versus 210.5±48.0 minutes, P<0.001), reduced post-CPVI fluoroscopic time (11.0±7.8 versus 13.7±8.9 minutes, P=0.006), and shorter energy delivery time (60.1±25.1 versus 75.0±24.3 minutes, P<0.001) were observed in the STABLE-SR group compared with the STEPWISE group. CONCLUSIONS: STABLE-SR is a simplified, personalized, and effective ablation strategy in nonparoxysmal AF patients. More importantly, over 50% nonparoxysmal AF patients do not need further ablation beyond CPVI and therefore can avoid excessive ablation. CLINICAL TRIAL REGISTRATION: URL: http://www.clinicaltrials.gov. Unique identifier: NCT01761188.