恩替卡韦与水飞蓟宾胶囊联合治疗慢性乙型肝炎患者临床效果分析

目的 探讨恩替卡韦与水飞蓟宾胶囊联合治疗慢性乙型肝炎患者的临床效果。 方法 2015年6月~2016年7月我院收治的140例慢性乙型肝炎患者,按照随机分组法将所有患者分为观察组（n=70)和对照组（n=70）。对照组患者口服恩替卡韦片治疗,在观察组,采用恩替卡韦片联合水飞蓟宾胶囊治疗。采用时间分辨免疫荧光法检测血清HBeAg,采用荧光定量PCR法检测血清HBV DNA,采用ELISA法检测血清白细胞介素-6 （IL-6）、TNF-α）和转化生长因子-β （TGF-β,使用FACSCalibur流式细胞仪检测外周血淋巴细胞亚群。 结果 在治疗24 w末,观察组临床症状如乏力、食欲不振、腹胀复常率分别为87.1%、90.0%、82.9%,显著高于对照组的41.4%、71.4%、58.6% （P<0.05）;观察组血清HBeAg和HBV DNA转阴率分别为0.0%和88.6%,与对照组的0.0%和68.6%比,无统计学差异（P>0.05）;观察组血清TBIL、ALT和AST水平分别为（16.49±3.17） μmol/L、（47.36±8.24） U/L和（40.58±5.26） U/L,显著低于对照组的（23.56±3.48） μmol/L、（68.25±7.93） U/L和（66.24±8.24） U/L（P<0.05）;观察组血清IL-6、TNF-α和TGF-β水平分别为（204.8±27.4） μg/L、（68.2±12.5） μg/L和（158.2±15.2） μg/L,显著低于对照组的（264.2±29.2） μg/L、（107.4±13.7） μg/L和（193.5±17.3） μg/L （P<0.05）;观察组外周血CD4+细胞百分比和CD4+/CD8+细胞比值分别为（40.36±3.61）%和（1.50±0.32）,显著高于对照组的【（33.46±3.17） %和（1.33±0.27）,P<0.05】。 结论 恩替卡韦片与水飞蓟宾胶囊联合治疗慢性乙型肝炎患者效果确切,可有效抑制HBV DNA复制,缓解机体炎症反应,提高患者的免疫功能。     

乙型肝炎肝硬化并发自发性细菌性腹膜炎患者血清白蛋白和肝纤维化指标的变化

目的 分析乙型肝炎肝硬化并发SBP患者血清肝纤维化指标及白蛋白水平的变化。方法 在我院2013年1月~2015年6月收治的31例乙型肝炎肝硬化并发自发性细菌性腹膜炎（SBP）患者和同期住院的31例乙型肝炎肝硬化未发生SBP患者,采用放射免疫法检测纤维化指标。结果 SBP组血清PCⅢ、Ⅳ-C、LN和HA水平分别为307.4±20.2μg/L、230.6±20.3μg/L、251.3±16.3μg/L、472.6±22.6μg/L,均显著高于非SBP组（269.8±16.5μg/L、181.6±17.9μg/L、215.6±13.3μg/L、436.1±20.3μg/L （均P<0.05）;SBP组血清和腹水ALB水平分别为23.3±2.2 g/L和10.0±1.3g/L,均显著低于非SBP组（28.9±2.7 g/L和15.9±1.5 g/L（均P<0.05）;SBP患者血清PCⅢ、Ⅳ-C、LN和HA水平均与ALB呈负相关（均P<0.05）。结论 乙型肝炎肝硬化患者并发SBP时可能影响血清肝纤维化指标水平。     

恩替卡韦联合自拟中药治疗慢性乙型肝炎患者疗效初步评价

目的 研究自拟中药联合恩替卡韦治疗慢性乙型肝炎患者的疗效以及不良反应发生情况。方法 2015年1月~2016年12月我科收治的69例慢性乙型肝炎患者,34例接受自拟中药（党参20 g,醋柴胡10 g,郁金10 g,鳖甲30 g,丹参20 g,龙葵10 g,白花蛇舌草15 g,绞股蓝15 g,三七10 g,片姜黄10 g,赤芍15 g,炙甘草6 g,水煎 400 ml,每日1剂,分早晚温服）联合恩替卡韦治疗,35例只接受恩替卡韦治疗。应用SPSS 19.0统计软件完成数据分析处理。结果 在治疗前和治疗24 w末,观察组与对照组血清ALB水平差别未见统计学意义（P> 0.05）,但治疗后观察组血清TBIL水平为（20.1± 4.7） μmol/L,显著低于对照组的（28.7± 4.8）μmol/L （P<0.05）;观察组血清ALT水平为（40.9± 6.8） U/L,显著低于对照组的（50.2± 6.7） U/L（P<0.05）;观察组血清AST水平为（39.6± 8.4） U/L,显著低于对照组的（52.5± 8.1） U/L （P<0.05）;治疗后,观察组和对照组血清透明质酸（HA）、层连蛋白（LN）、4型胶原（IV-C）和3型前胶原（PC-Ⅲ）水平均有所下降,其中观察组血清HA水平由[（256.3± 9.3）μg/L下降至（98.3± 9.1） μg/L,对照组血清HA水平由[（255.3± 9.8） μg/L下降至（140.3± 9.6） μg/L,观察组血清LN水平由[（241.3± 7.8） μg/L下降至（110.3± 7.7） μg/L,对照组血清LN水平由[（240.4± 7.8） μg/L下降至（170.5± 7.9） μg/L,观察组血清IV-C水平由[（98.4± 6.8） μg/L降至（53.6± 6.7） μg/L,对照组血清IV-C水平由[（98.00± 6.5） μg/L下降至（70.9± 6.6） μg/L,观察组血清PC-Ⅲ水平由[（193.6± 13.2） μg/L下降至（98.3± 9.9） μg/L,对照组血清PC-Ⅲ水平由[（193.5± 13.4） μg/L下降至（120.3± 9.7） μg/L,显示出治疗后观察组肝纤四项指标均显著低于对照组（P<0.05）。另外,治疗后观察组与对照组血清HBeAg转阴率和HBV DNA转阴率均未见统计学差异,而消化道症状,如腹泻、腹痛以及呕吐的发生率在观察组和对照组分别为2.9%和8.6%,差别也无统计学意义（P>0.05）。结论 自拟中药联合恩替卡韦联合治疗慢性乙型肝炎患者,两者结合一方面可以迅速抑制HBV复制,在另一方面可以发挥祖国医学治本的优势,扶正而固本,增强了疗效。     

慢性乙型肝炎肝衰竭患者血清甲胎蛋白和胸腺素β4水平变化

目的 探讨血清甲胎蛋白（AFP）和胸腺素β4水平在预测慢性乙型肝炎肝衰竭患者转归中的应用价值。方法 2015年1月~2016年7月我院收治的慢性乙型肝炎肝衰竭患者75例,采用ELISA法检测血清AFP和胸腺素β4水平。结果 治疗3个月,在75例慢性乙型肝炎肝衰竭患者中,43例患者（57.3%）死亡,32例患者（42.7%）生存;入院时,生存组血清AFP和胸腺素β4水平分别为（194.2±13.5） μg/L和（914.1±29.5） μg/L,均显著高于死亡组的【（98.4±9.5） μg/L和（504.2±15.4） μg/L,P﹤0.05】;以血清AFP大于或等于71.3 μg/L为截断点,其预测患者在3个月内生存的敏感度为81.5%,特异度为74.9%;以血清胸腺素β4大于472.7μg/L为截断点,其预测患者在3个月内生存的敏感度为76.2%,特异度为69.5%。结论 慢性乙型肝炎肝衰竭患者血清AFP和胸腺素β4水平对患者转归有一定的预测作用,两者水平越高,预后越好。     

乙型肝炎肝硬化患者血清血小板衍生生长因子和叶酸水平变化

目的 分析乙型肝炎肝硬化患者血清血小板衍生生长因子（PDGF）和叶酸水平变化。方法 2015年1月~2016年12月期间我院收治的110例乙型肝炎肝硬化患者,常规行肝功能Child-Pugh分级,采用ELISA法检测血清PDGF水平,采用化学发光法检测血清叶酸水平,采用放射免疫法检测血清Ⅲ型前胶原（PCⅢ）、Ⅳ型胶原（Ⅳ-C）、层粘连蛋白（LN）和透明质酸酶（HA）水平。结果 32例Child-Pugh C级患者血清PDGF水平为（190.7±11.4）pg/ml,显著高于42例B级或36例A级患者的（128.5±8.1） pg/ml或（79.5±4.9） pg/ml（P<0.05）;Child-Pugh C级患者血清叶酸水平为（2.7±0.5） μg/L,显著低于B级或A级患者的（5.3±0.5） μg/L或（7.1±0.8） μg/L（P<0.05）;Child-Pugh C级患者血清谷丙转氨酶（ALT）、总胆红素（TBIL）水平和INR水平分别为（102.4±6.9） U/L、（60.7±5.8） μmol/L和（1.9±0.2）,均显著高于B级患者【分别为（78.9±4.5） U/L、（36.4±3.2） μmol/L和（1.5±0.2）】或A级患者[分别为（56.8±3.9） U/L、（26.8±2.8） μmol/L和（1.2±0.1）,均P<0.05】;Child-Pugh C级患者血清白蛋白（ALB）为（23.9±2.4） g/L,显著低于B级患者的（30.8±2.7） g/L或A级患者的（41.0±3.3） g/L（均P<0.05）;Child-Pugh C级患者血清PCⅢ、Ⅳ-C、LN和HA水平分别为（279.7±19.2） μg/L、（140.4±8.0） μg/L、（159.7±8.2） μg/L和（237.8±9.1） μg/L,均显著高于B级分别为【（238.5±17.8） μg/L、（94.8±6.2） μg/L、（129.5±7.3） μg/L和（174.1±8.6） μg/L或A级患者（分别为158.6±13.6）μg/L、（75.4±5.1） μg/L、（96.8±6.8） μg/L和（128.3±6.9）μg/L,均P<0.05】;肝硬化患者血清PDGF水平与ALT、TBIL、INR和肝纤维化指标均呈正相关,与ALB呈负相关（均P<0.05）,叶酸水平与ALT、TBIL、INR和肝纤维化指标均呈负相关,与ALB呈正相关（均P<0.05）。结论 随着肝功能损害程度的加重,肝硬化患者血清PDGF水平逐渐升高,而叶酸水平逐渐降低,其临床意义还有待于进一步研究。     

恩替卡韦联合复方鳖甲软肝片治疗乙型肝炎肝硬化患者疗效及转归研究*

目的 研究应用恩替卡韦（ETV）联合复方鳖甲软肝片治疗乙型肝炎肝硬化患者的疗效及其对疾病转归的影响。方法 2014年1月~2016年12月我院收治的乙型肝炎肝硬化患者76例,采用随机数字表法将患者分为对照组36例和观察组40例,分别给予ETV或ETV联合复方鳖甲软肝片治疗,观察12 m月。采用 PCR法定量检测血清HBV DNA载量,采用放射免疫法检测血清透明质酸（HA）、Ⅲ型前胶原（PⅢP）、层粘连蛋白（LN）和Ⅳ型胶原（CⅣ）,常规行血生化和Fibroscan检查。结果 在治疗12 m末,观察组血清TBIL为（19.7±13.4） μmol/L,ALT水平为（28.3±9.4） U/L,均显著低于对照组的[（23.2±13.8） μmol/L和（53.6±10.2） U/L,P<0.05],观察组血清ALB水平为（37.2±5.6） g/L,PTA为（91.1±11.2） %,显著高于对照组[（34.7±6.1） g/L和（81.4±10.5） %,P<0.05];观察组CTP评分为（6.3±1.2）,显著低于对照组的[（8.1±1.4）,P<0.05],肝脏硬度检测（LSM）为（16.5±12.2） kPa,也显著低于对照组的[（22.7±14.4） kPa,P<0.05];观察组血清HA水平为（89.2±43.1） μg/L,PⅢP为（119.7±60.8） μg/L,LN为（98.7±30.2） μg/L,CⅣ为（102.6±29.7） μg/L,均低于对照组[分别为（184.3±58.2） μg/L、（254.5±74.7） μg/L、（140.8±39.7） μg/L、（165.7±41.2） μg/L,P<0.05];观察组血清HBV DNA阴转率为97.5%（39/40）,与对照组的94.4%（34/36）比,差异无统计学意义（P>0.05）,观察组血清HBV DNA载量为（3.2±0.3）log10 copies/ml,与对照组的（3.5±0.4）log10 copies/ml比,无显著性差异（P>0.05）;观察组腹水、肝性脑病、消化道出血和肝细胞癌发生率分别为17.5%、5.0%、10.0%和2.5%,显著低于对照组的52.7%、13.8%、16.7%和11.1% （P<0.05）;在观察的12 m内,观察组无死亡病例,而对照组死亡3例（8.3%）。结论 应用ETV联合复方鳖甲软肝片治疗能够明显改善乙型肝炎肝硬化患者肝功能指标,降低血清肝纤维化指标,改善预后。     

原发性肝癌患者TACE术后血清HIF－1α和VEGF水平变化

目的调查原发性肝癌（PLC）患者在经肝动脉化疗栓塞术（TACE）后血清乏氧诱导因子-1α（HIF-1α）和血管内皮生长因子（VEGF）水平变化及其与疗效的关系。方法2014年1月~2015年12月我院接受治疗的原发性肝肝癌（PLC）患者58例,在TACE术前后,采用ELISA法检测血清HIF－1α和VEGF水平。使用CT行腹部检查,采用实体瘤疗效考核标准考核疗效。结果在TACE术后3 m,在58例PLC患者中,获得完全缓解、部分缓解、疾病稳定和疾病进展者分别为10例（17.2%）、15例（25.9%）、20例（34.5%）和13例（22.4%）;10例完全缓解患者术后1 m血清dHIF-1α（147.52±28.69 μg/L）和VEGF水平（257.69±47.86 μg/L）显著低于48例未获得完全缓解患者【分别为（330.12±39.52 μg/L）和（402.74±96.25 μg/L）,P<0.05】;术后3 m完全缓解患者血清HIF-lα和VEGF水平分别为（109.89±11.54 μg/L和177.47±79.46 μg/L）,显著低于未获得完全缓解患者【分别为（276.34±54.83 μg/L和380.70±134.85 μg/L）,P<0.05】;在TACE术前后PLC患者血清AFP水平也呈类似变化。结论血清HIF-1α和VEGF水平与肿瘤血管的形成有关,在TACE术后获得完全缓解的PLC患者血清水平降低,可能成为考核疗效的重要指标。     

恩替卡韦治疗乙型肝炎肝硬化伴肝源性糖尿病患者疗效及其对糖代谢指标的影响

目的 探讨恩替卡韦治疗乙型肝炎肝硬化并发肝源性糖尿病（HD）患者疗效及其对糖代谢指标的影响。方法 采用随机数字表法将84例乙型肝炎肝硬化并发HD患者分为对照组42例和观察组42例,分别给予降糖、护肝等常规治疗或加用恩替卡韦抗病毒治疗,观察48 w。观察的糖代谢指标包括空腹血糖（FBG）、餐后2 h血糖 （2h PBG）、糖化血红蛋白（HbA1c）、空腹胰岛素（FINS）和胰岛素抵抗指数（HOMA-IR）。采用放射免疫法测定血清肝纤维化指标,如层黏连蛋白（LN）、血清透明质酸（HA）、Ⅲ型前胶原（PCⅢ）、Ⅳ型胶原（Ⅳ-C）。结果 在治疗48 w末,观察组33例（78.6%）患者出现病毒学应答,对照组仅7例（16.7%）出现自发血清HBV DNA阴转,两组差异有统计学意义（P<0.05）;在治疗48 w末,观察组血清TBIL、ALT、ALB水平分别为（21.2±4.2） μmol/L、（43.3±12.7） IU/L和（39.4±1.8） g/L,与对照组的【（36.1±5.3） μmol/L、（77.1±15.2） IU/L和（33.9±3.4） g/L比,差异具有统计学意义（P<0.05）;FBG、2h PG、HbA1c、FINS和HOMA-IR水平分别为（5.3±0.7） mmol/L、（7.1±1.5） mmol/L、（6.1±0.6） %、（10.5±2.6） mU/L和（2.1±0.7）,显著低于对照组【（6.8±0.8） mmol/L、（10.3±1.6） mmol/L、（7.8±0.7） %、（15.6±4.0） mU/L和（3.1±0.6）,P<0.05】;血清LN、HA、PCⅢ、Ⅳ-C水平分别为（144.3±54.2） μg/L、（81.7±38.7） μg/L、（116.3±31.4） μg/L和（71.2±42.9） μg/L,显著低于对照组【（177.3±63.5） μg/L、（123.1±41.9） μg/L、（165.7±28.6） μg/L和（124.7±39.1） μg/L,P<0.01】。结论 恩替卡韦治疗乙型肝炎肝硬化并发HD患者可抑制HBV复制,改善肝功能,稳定HD病情。     

干扰素α-2b与聚乙二醇干扰素α-2a治疗慢性乙型肝炎患者疗效比较研究*

目的 观察干扰素α-2b与聚乙二醇干扰素α-2a（PEG-IFNα-2a）治疗慢性乙型肝炎（CHB）患者的临床疗效。方法 将116例CHB患者分成两组,每组58例。给予对照组普通干扰素α-2b治疗,给予观察组PEG-IFNα-2a,疗程均为48 w。检测血清层黏连蛋白（LN）、透明质酸（HA）、Ⅳ型胶原（Ⅳ-C）、Ⅲ型前胶原肽（PⅢP）及血清白介素4（IL-4）、白介素6（IL-6）、白介素10（IL-10）和干扰素γ（IFN-γ）变化。结果 在治疗结束时,观察组HBV DNA转阴率、HBeAg转阴率、HBeAg血清转换率和ALT复常率分别为75.9%、29.3%、22.4%和70.7%,显著高于对照组的51.7%、19.0%、10.3%和55.2%（P<0.05）;观察组血清LN为（148.5±46.0） μg/L,显著低于对照组【（162.6±26.7） μg/L,P<0.05】,血清HA为（158.7±67.9）μg/L,显著低于对照组【（201.4±55.1） μg/L,P<0.05】,血清Ⅳ-C为（108.3±33.4） μg/L,显著低于对照组【（119.2±62.4） μg/L,P<0.05】,血清PⅢP为（145.8±47.2） μg/L,显著低于对照组【（155.3±36.7）μg/L,P<0.05】;观察组血清IL-4水平为（1.5±0.6） pg/ml,显著低于对照组【（3.9±1.6） pg/ml,P<0.05】,而IL-6水平为（6.9±1.3） pg/ml,显著高于对照组【（3.6±0.9） pg/ml,P<0.05】,IL-10水平为（22.6±13.4） pg/ml,显著低于对照组【（17.3±11.4） pg/ml,P<0.05】,IFN-γ水平为（37.2±10.2） pg/ml,显著高于对照组【（28.3±10.5） pg/ml,P<0.05】;观察组血小板计数减少发生率为51.7%,显著高于对照组的12.1%（P<0.05）。结论 PEG-IFNα-2a治疗CHB患者临床疗效优于普通干扰素α-2b,能持久有效地抑制HBV复制,改善血清肝纤维化指标,但不良反应较大,应及时处理。     

肝硬化患者胃粘膜病变与血清胃蛋白酶原水平变化

目的 探讨肝硬化合并胃黏膜病变患者血清胃蛋白酶原（PG）水平的变化情况。方法 2011年9月~2014年3月我院收治的肝硬化患者,经胃镜检查发现胃黏膜正常138例,并发胃黏膜病变156例,后者又分为糜烂性胃炎组56例、萎缩性胃炎组37例、胃溃疡组23例、十二指肠球部溃疡组22例和胃癌组18例。采用时间分辨荧光免疫分析法测定血清PG水平。结果 肝硬化合并胃黏膜病变患者血清PGI 水平、PGI/PGII比值[（78.08±21.33）μg/L,（6.75±1.81）]高于无胃粘膜病变组[（51.37±19.85） μg/L,（4.87±1.56）,差异有统计学意义（P<0.05）];两组血清PGII水平[（12.16±5.60）μg/L 对 （13.02±4.96）μg/L]比较,差异无统计学意义（P>0.05）;萎缩性胃炎组和胃癌组血清PGI水平[分别为（34.18±11.35）μg/L和（28.96±12.14）μg/L]显著低于糜烂性胃炎组[（108.28±20.87）μg/L]、胃溃疡组[（113.37±20.04）μg/L]、十二指肠球部溃疡组[（138.65±19.38）μg/L],差异有统计学意义（P<0.05）。结论 肝硬化患者血清PG水平与胃黏膜病变密切相关,不同类型胃病患者血清PG水平不同,对临床有重要的参考价值。     

慢性乙型肝炎患者肝细胞内乙型肝炎核心抗原阳性的临床意义

目的探讨CHB患者肝组织HBcAg阳性的意义。方法对200例CHB患者应用荧光聚合酶链反应（FQ-PCR）法精确定量检测血清HBV DNA含量。患者均检测血清中HBeAg含量,同时进行肝活组织检查,应用免疫组织化学技术检测HBcAg情况,并进行相关性分析。结果按测定血清HBV DNA水平,分为A组（＜3 log10拷贝/ml）20例,B组（≥3 log10拷贝/ml-＜5 log10拷贝/ml）13例,C组（≥5 log10拷贝/ml～＜6 log10拷贝/ml）24例,D组（≥6 log10拷贝/ml～＜8 log10拷贝/ml）116例,E组（≥8 log10拷贝/ml）27例。肝组织HBcAg阳性者175例,占87．5％,A组HBcAg阳性率55．0％（11/20）,B组53．8％（7/13）,C组75．0％（18/24）,D组96．6％（112/116）,E组100．0％（27/27）,HBcAg阳性率与血清HBV DNA水平之间呈显著正相关（r=0．80,P＜0．01）。血清HBV DNA水平高低与HBeAg阳性率之间呈显著正相关（r=0．47,P＜0．01）。其中20例HBV DNA阴性者中（A组）,HBeAg阳性者5例（25％）,HBcAg阳性者11例（55％）；15例HBV DNA阴性且HBeAg阴性者中有7例HBcAg阳性,占46．7％。结论CHB患者肝组织HBcAg阳性能更可靠地反映肝细胞内HBV复制状态。检测肝组织内HBcAg对CHB患者疗效评价和对治疗反应性的预测更具有临床意义。     

一种新的HBeAg阴性慢性乙型肝炎病毒变异机制

Objective To investigate mutation patterns in core promoter(CP)region of hepatitis B virus(HBV).Methods HBV DNA was extracted from sera of patients with chronic HBV infection.The CP sequence was amplified by polymerase chain reaction(PCR)and cloned into pMD19 T vector.The positive clones were then sequenced.The sequences were compared with known HBV genome in GenBank to identify the mutation sites and patterns of patients with chronic HBV infection.Results There were 74 clones from 21 patients with chronic HBV infection which were sequenced.The sequence comparisons showed that there was a 234-nucleotide deletion in CP region of HBV genome in 54 clones and a 245-nucleotide deletion in one clone.These deletion regions included CP,HBeAg initiation codon and direct repeat sequence(DR)Ⅰ regions,which named CP deletion(CPD).A1585T replacement mutation was also found in HBV strain with CPD,which indicated that there was linkage between these two mutations.Conclusions A novel mechanism of HBeAg negative chronic hepatitis B is observed,which includes deletions of CP and HBeAg initiation codon.Meanwhile,a simple and useful PCR method is developed to detect CPD.     

一种新的HBeAg阴性慢性乙型肝炎病毒变异机制

Objective To investigate mutation patterns in core promoter(CP)region of hepatitis B virus(HBV).Methods HBV DNA was extracted from sera of patients with chronic HBV infection.The CP sequence was amplified by polymerase chain reaction(PCR)and cloned into pMD19 T vector.The positive clones were then sequenced.The sequences were compared with known HBV genome in GenBank to identify the mutation sites and patterns of patients with chronic HBV infection.Results There were 74 clones from 21 patients with chronic HBV infection which were sequenced.The sequence comparisons showed that there was a 234-nucleotide deletion in CP region of HBV genome in 54 clones and a 245-nucleotide deletion in one clone.These deletion regions included CP,HBeAg initiation codon and direct repeat sequence(DR)Ⅰ regions,which named CP deletion(CPD).A1585T replacement mutation was also found in HBV strain with CPD,which indicated that there was linkage between these two mutations.Conclusions A novel mechanism of HBeAg negative chronic hepatitis B is observed,which includes deletions of CP and HBeAg initiation codon.Meanwhile,a simple and useful PCR method is developed to detect CPD.     

一种新的HBeAg阴性慢性乙型肝炎病毒变异机制

Objective To investigate mutation patterns in core promoter(CP)region of hepatitis B virus(HBV).Methods HBV DNA was extracted from sera of patients with chronic HBV infection.The CP sequence was amplified by polymerase chain reaction(PCR)and cloned into pMD19 T vector.The positive clones were then sequenced.The sequences were compared with known HBV genome in GenBank to identify the mutation sites and patterns of patients with chronic HBV infection.Results There were 74 clones from 21 patients with chronic HBV infection which were sequenced.The sequence comparisons showed that there was a 234-nucleotide deletion in CP region of HBV genome in 54 clones and a 245-nucleotide deletion in one clone.These deletion regions included CP,HBeAg initiation codon and direct repeat sequence(DR)Ⅰ regions,which named CP deletion(CPD).A1585T replacement mutation was also found in HBV strain with CPD,which indicated that there was linkage between these two mutations.Conclusions A novel mechanism of HBeAg negative chronic hepatitis B is observed,which includes deletions of CP and HBeAg initiation codon.Meanwhile,a simple and useful PCR method is developed to detect CPD.     

一种新的HBeAg阴性慢性乙型肝炎病毒变异机制

Objective To investigate mutation patterns in core promoter(CP)region of hepatitis B virus(HBV).Methods HBV DNA was extracted from sera of patients with chronic HBV infection.The CP sequence was amplified by polymerase chain reaction(PCR)and cloned into pMD19 T vector.The positive clones were then sequenced.The sequences were compared with known HBV genome in GenBank to identify the mutation sites and patterns of patients with chronic HBV infection.Results There were 74 clones from 21 patients with chronic HBV infection which were sequenced.The sequence comparisons showed that there was a 234-nucleotide deletion in CP region of HBV genome in 54 clones and a 245-nucleotide deletion in one clone.These deletion regions included CP,HBeAg initiation codon and direct repeat sequence(DR)Ⅰ regions,which named CP deletion(CPD).A1585T replacement mutation was also found in HBV strain with CPD,which indicated that there was linkage between these two mutations.Conclusions A novel mechanism of HBeAg negative chronic hepatitis B is observed,which includes deletions of CP and HBeAg initiation codon.Meanwhile,a simple and useful PCR method is developed to detect CPD.     

一种新的HBeAg阴性慢性乙型肝炎病毒变异机制

Objective To investigate mutation patterns in core promoter(CP)region of hepatitis B virus(HBV).Methods HBV DNA was extracted from sera of patients with chronic HBV infection.The CP sequence was amplified by polymerase chain reaction(PCR)and cloned into pMD19 T vector.The positive clones were then sequenced.The sequences were compared with known HBV genome in GenBank to identify the mutation sites and patterns of patients with chronic HBV infection.Results There were 74 clones from 21 patients with chronic HBV infection which were sequenced.The sequence comparisons showed that there was a 234-nucleotide deletion in CP region of HBV genome in 54 clones and a 245-nucleotide deletion in one clone.These deletion regions included CP,HBeAg initiation codon and direct repeat sequence(DR)Ⅰ regions,which named CP deletion(CPD).A1585T replacement mutation was also found in HBV strain with CPD,which indicated that there was linkage between these two mutations.Conclusions A novel mechanism of HBeAg negative chronic hepatitis B is observed,which includes deletions of CP and HBeAg initiation codon.Meanwhile,a simple and useful PCR method is developed to detect CPD.     

一种新的HBeAg阴性慢性乙型肝炎病毒变异机制

Objective To investigate mutation patterns in core promoter(CP)region of hepatitis B virus(HBV).Methods HBV DNA was extracted from sera of patients with chronic HBV infection.The CP sequence was amplified by polymerase chain reaction(PCR)and cloned into pMD19 T vector.The positive clones were then sequenced.The sequences were compared with known HBV genome in GenBank to identify the mutation sites and patterns of patients with chronic HBV infection.Results There were 74 clones from 21 patients with chronic HBV infection which were sequenced.The sequence comparisons showed that there was a 234-nucleotide deletion in CP region of HBV genome in 54 clones and a 245-nucleotide deletion in one clone.These deletion regions included CP,HBeAg initiation codon and direct repeat sequence(DR)Ⅰ regions,which named CP deletion(CPD).A1585T replacement mutation was also found in HBV strain with CPD,which indicated that there was linkage between these two mutations.Conclusions A novel mechanism of HBeAg negative chronic hepatitis B is observed,which includes deletions of CP and HBeAg initiation codon.Meanwhile,a simple and useful PCR method is developed to detect CPD.     

一种新的HBeAg阴性慢性乙型肝炎病毒变异机制

Objective To investigate mutation patterns in core promoter(CP)region of hepatitis B virus(HBV).Methods HBV DNA was extracted from sera of patients with chronic HBV infection.The CP sequence was amplified by polymerase chain reaction(PCR)and cloned into pMD19 T vector.The positive clones were then sequenced.The sequences were compared with known HBV genome in GenBank to identify the mutation sites and patterns of patients with chronic HBV infection.Results There were 74 clones from 21 patients with chronic HBV infection which were sequenced.The sequence comparisons showed that there was a 234-nucleotide deletion in CP region of HBV genome in 54 clones and a 245-nucleotide deletion in one clone.These deletion regions included CP,HBeAg initiation codon and direct repeat sequence(DR)Ⅰ regions,which named CP deletion(CPD).A1585T replacement mutation was also found in HBV strain with CPD,which indicated that there was linkage between these two mutations.Conclusions A novel mechanism of HBeAg negative chronic hepatitis B is observed,which includes deletions of CP and HBeAg initiation codon.Meanwhile,a simple and useful PCR method is developed to detect CPD.     

一种新的HBeAg阴性慢性乙型肝炎病毒变异机制

Objective To investigate mutation patterns in core promoter(CP)region of hepatitis B virus(HBV).Methods HBV DNA was extracted from sera of patients with chronic HBV infection.The CP sequence was amplified by polymerase chain reaction(PCR)and cloned into pMD19 T vector.The positive clones were then sequenced.The sequences were compared with known HBV genome in GenBank to identify the mutation sites and patterns of patients with chronic HBV infection.Results There were 74 clones from 21 patients with chronic HBV infection which were sequenced.The sequence comparisons showed that there was a 234-nucleotide deletion in CP region of HBV genome in 54 clones and a 245-nucleotide deletion in one clone.These deletion regions included CP,HBeAg initiation codon and direct repeat sequence(DR)Ⅰ regions,which named CP deletion(CPD).A1585T replacement mutation was also found in HBV strain with CPD,which indicated that there was linkage between these two mutations.Conclusions A novel mechanism of HBeAg negative chronic hepatitis B is observed,which includes deletions of CP and HBeAg initiation codon.Meanwhile,a simple and useful PCR method is developed to detect CPD.     

一种新的HBeAg阴性慢性乙型肝炎病毒变异机制

Objective To investigate mutation patterns in core promoter(CP)region of hepatitis B virus(HBV).Methods HBV DNA was extracted from sera of patients with chronic HBV infection.The CP sequence was amplified by polymerase chain reaction(PCR)and cloned into pMD19 T vector.The positive clones were then sequenced.The sequences were compared with known HBV genome in GenBank to identify the mutation sites and patterns of patients with chronic HBV infection.Results There were 74 clones from 21 patients with chronic HBV infection which were sequenced.The sequence comparisons showed that there was a 234-nucleotide deletion in CP region of HBV genome in 54 clones and a 245-nucleotide deletion in one clone.These deletion regions included CP,HBeAg initiation codon and direct repeat sequence(DR)Ⅰ regions,which named CP deletion(CPD).A1585T replacement mutation was also found in HBV strain with CPD,which indicated that there was linkage between these two mutations.Conclusions A novel mechanism of HBeAg negative chronic hepatitis B is observed,which includes deletions of CP and HBeAg initiation codon.Meanwhile,a simple and useful PCR method is developed to detect CPD.