Result: We obtained about 9.0 million 32-mer short reads on average per sample, and mapping rates to miRBase were 15.5%. In the statistical analysis, the p-value and the expression levels of 110 miRNAs were found to be differentially expressed in the 3 groups. 16 miRNAs

were up-regulated in CH-B patients with miR-3591-5p being the most enriched. To set up the condition of miRNA-mRNA pairings with perfect matching of the seed region, RNAhybrid 2.2 analysis predicted that human hepatic cells might use 8 out of 16 miRNAs to down regulate the expression of HBV P and S genes. Then, eight HBV genomic segments containing putative target sites for human miRNAs were separately cloned in the psiCheck-2 vector. The HBV genomic segment predicted to be targeted by miR-125b-5p inhibited Saracatinib research buy remarkably the expression of the reporter in HepG2 and Huh-7 cells. Transfection of miR-125b-5p mimic in HepG2 cells increased the reporter silencing effect.

Moreover, LBH589 transfection of the inhibitor in the cells reduced the reporter silencing activity. Conclusion: We demonstrated that 16 miRNAs were up-regulated in patients with HBV chronic infection. miR-125b-5p, one of the 16 miRNAs, may be responsible for the silencing effect on the HBV genome segment. Disclosures: The following people have nothing to disclose: Masashi Ninomiya, Yasuteru Kondo, Takayuki Kogure, Eiji Kakazu, Osamu Kimura, Tatsuki Morosawa, Tomoaki Iwata, Yasuyuki Fujisaka, Tooru Shimosegawa Background and Aims: Long-term treatment with tenofovir (TDF) is effective in suppressing viral replication in HBeAg-ve and +ve chronic hepatitis B (CHB) patients, but loss of HBsAg is rare. The effect of stopping TDF in a CHB patient with long-term HBV-DNA suppression or HBsAg loss is uncertain. Methods: Among 25 TDF-treated CHB patients with persistently undetectable HBV-DNA for a median time of 7.46 years, therapy was stopped in 7 prospectively followed-up patients. Hepatitis B virus (HBV) quasispecies between codons rt163-rt278 was studied

by ultra-deep pyrosequencing MCE公司 (UDPS, GS-FLX/Junior, Roche) in patients in whom amplification of HBV-DNA at baseline (BA) and after TDF discontinuation (Post) was possible. Results: At BA, median HBV-DNA and HBsAg levels (qHBsAg) were 5.61 (4.82-8.59) and 3.33 (1.95-5.31) logIU/mL respectively, median age 54.81 (43.07-62.86) years, HBV genotype A 1, D 4 and F 1. At end of treatment, all patients were HBeAg-ve, HBV-DNA undetectable, and median qHBsAg was 2.99 (2.02-3.70) logIU/mL. After stopping TDF fora median of 7.29 weeks, no patient cleared HBsAg and HBV-DNA was detectable in all cases (median, 4.52 [1.75-5.34] logIU/mL) with no changes in ALT or qHBsAg. UDPS analysis of HBV quasispecies in 3 patients showed no changes in the master sequence between BA and Post despite 7 years’ complete suppression of HBV replication, but low-frequency variants between positions rtG21 0 and rtS238 were detected at Post: 1 patient showed variants rtV214A (0.26%), rtQ215S (0.