| Objective To investigate of HBV quasispecies evolution in the relapse mechanism for the chronic hepatitis B patients with lamivudine treatment-na ve.Methods43CHB patients including25LAM initial treatment cases and18retreatment cases stopped treatement when they had reached the endpointcriteria of2008APASL guideline or after extending the period of treatment. Wecollected patients’ baseline serum, standard withdrawal for recurrent liver tissueand serum. Laboratory methods: serum rcDNA were extracted by water boiling,hepatocyte full-length HBV cccDNA were amplified with HBV cccDNA specialprimers Rolling circle amplification (RCA); mutations at virologicalbreakthrough, drug withdrawal, relapse of polymerase(P)gene were amplifiedand cloned sequenced and analyzed by bioinformatics software. We analysis HBV rcDNA of serum before treatement, treatment for recurrent HBV cccDNAof liver cells and serum HBV rcDNA P gene characteristics and recurrence.Results (1)The P mutation of N/H238T(14/43)were the most highest frequency in The baseline of serumHBV rcDNA;the P mutation of D134E/I（9/16）、L/F/Q/R267H(6/16)、N/H238T(6/16)、 V278T(6/16)、 T222A(6/16)was the most highest frequency in Initial treatment group, and the most highest frequency in retreatment group was N/H238T(8/14)、L/F/Q/R267H(6/14)、V278T(6/14)、T222A(3/14); The P mutation of L/F/Q/R267H (25/43)were the most highest frequency in stopping LAM hepatocyte cccDNA, the cccDNA P mutation of D134E/I (12/25)、L/F/Q/R267H（10/25）、V278T（8/25）、N/H238T（7/25）was the most highest frequency in Initial treatment group, and the most highest frequency in retreatment group was L/F/Q/R267H(15/18)、V278T（10/18）、N/H238T（9/18）, The P mutation ofL/F/Q/R267H (25/43) were the most highest frequency in replased N/H238T(12/19)、 L/F/Q/R267H(10/19)、T222A(5/19), the P mutation of N/H238T(6/10)、 L/F/Q/R267H(5/10)、T222A(5/10)was the most highest frequency in Initial treatment group, and the most highest frequency in retreatmentgroup was N/H238T(6/9)、V278T(6/9)、L/F/Q/R267H(5/9)、T222A(5/9);(2)The T222A mutation frequency of serumHBV rcDNA in relapse patientwas higher than no relapse patient In The baseline(p<0.05), Liver cells did not relapse patients cccDNA P/F/L Q/R267H, D134E/I detection rate higher than that of patients with recurrence (P <0.05), the initial set of liver cells did not relapse patients cccDNA P/F/L Q/R267H, D134E/I detection rate higher than that of patients with recurrence (P <0.05),and The N/H238T mutation frequency of hepatocyte cccDNA in relapse patient wa s higher than no relapse patient (p<0.05);(3) The quasispecies complexityof hepatocyte cccDNA in relapse patient was higher than no relapse patient at stopped NUCs(p<0.05), The quasispecies complexity of hepatocytecccDNA in relapse patient was higher than no relapse patient in retreatment group at stopped NUCs(p<0.05);(4)The quasispecies complexity of hepatocyte cccDNA in retreatment group patient was higher than in Initial treatment grou at stopped NUCs (p<0.05).(5) For recurrent relapse group ofquasi kinds of complexity:>> baseline standard drug withdrawal, no recurrence quasi kinds of complexity: standard drug withdrawal when <treatment at baseline.Conclusion (1) The mutation of T222A and N/H238T may be associatedwith relapse after drug withdrawal.(2) Treatment of baseline HBV P gene DNA area V278T mutation frequency, the higher the risk of relapse.(3)The quasispecies complexity of serum HBV rcDNA and hepatocyte cccDNA may be associated with relapse after drug withdrawal.(4) Withdrawal when liver cells cccDNA P area kind of complexity is higher the more easy to relapse.(5) D134E/I mutation frequency is higher, the more itis not easy to relapse... |
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