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目的就拉米夫定治疗慢性乙型肝炎(C H B)1a的疗效、H BV D N A P基因变异特点及编码的蛋白质的二级结构加以研究。方法提取H BV D N A后,经半巢式聚合酶链反应(PC R)扩增,PC R产物自动测序。总结拉米夫定的疗效、P基因变异出现的时间,用SAS软件分析影响疗效的因素。用D N A ST AR软件的C LU STA L V方法对H BV D N A P基因片段的核苷酸和氨基酸差异、变异类型进行分析。用D N Aclub、D N Asis软件分析出现P基因变异前后所编码的蛋白质的二级结构的差异。结果按照ALT水平分组,各组间促进A LT正常化没有明显差异。在治疗后6个月血清标本中未发现Y M D D变异,在治疗后12个月血清标本中发现2例Y M D D变异。在对照组中发现了1例L528M变异;在治疗后12个月发现了2例Y V D D变异,还发现了2例只有L528M的变异。在发生变异后,Y M D D的转角结构变为折叠,L528M的转角结构没变。结论1a的拉米夫定治疗可以有效抑制H BV D N A的复制。Y M D D变异发生在拉米夫定治疗6个月之后,L528M变异可单独在体内存在,未接受抗病毒治疗的慢性H BV携带者可发生L528M变异。发生Y M D D变异的P基因编码的蛋白质二级结构由转角变为折叠。L528M变异不影响蛋白质二级结构。
Objective To investigate the efficacy of lamivudine in the treatment of chronic hepatitis B (C H B) 1a, the variation of H BV D N A P gene and the secondary structure of the encoded protein. Methods After extracting H BV D N A, PC R products were amplified by semi-nested polymerase chain reaction (PCR) and sequenced automatically. Summarizes the efficacy of lamivudine, P gene mutation appeared time, with SAS software analysis of the factors that affect the efficacy. The nucleotide and amino acid differences and mutation types of H BV D N A P gene fragment were analyzed by C LU STA L V method of D N A ST AR software. Differences in the secondary structure of the proteins encoded before and after the P gene mutation were analyzed using D N Aclub, D N Asis software. The results were grouped according to the level of ALT. There was no significant difference between groups in promoting the normalization of A LT. No YMDD mutation was found in serum samples at 6 months after treatment, and 2 cases of YMDD mutation were found in serum samples at 12 months after treatment. One case of L528M mutation was found in the control group; two cases of YV D D mutation were found at 12 months after treatment and two cases of only L528M mutation were found. After mutating, the corner structure of Y M D D becomes folded, and the corner structure of L528 M does not change. Conclusions 1a lamivudine treatment can effectively inhibit the replication of H BV D N A. Y M D D Variation After 6 months of lamivudine treatment, the L528M variant may exist alone in the body, and the L528M mutation may occur in chronic H BV carriers who have not received antiviral therapy. The secondary structure of the protein encoded by the P gene with the Y M D D mutation changes from a turn to a fold. L528M mutation does not affect protein secondary structure.