慢性乙型肝炎患者丙氨酸氨基转移酶水平与HBV DNA转阴的相关性

目的 观察慢性乙型肝炎患者ALT高水平与HBV DNA转阴的相关性,以探讨抗病毒治疗CHB患者的时机,为更加合理应用抗病毒药物提供参考.方法 应用回顾性分析的方法,对本院就诊的HBV DNA＞1×104拷贝/ml且ALT＞800 U/L的177例CHB患者,分为未抗病毒组96例,抗病毒组81例,在4、8、12、24周,各检测1次HBV DNA、HBV标志物及ALT,随访时间为24～228周.两组间计量资料方差齐时采用t检验,率的比较采用x2检验或Fisher's确切概率法.结果 随访24周者,未抗病毒组71例,抗病毒组65例.24周内未抗病毒组和抗病毒组HBV DNA阴转者分别为62例(87.3%)和63例(96.9%);8周内HBV DNA阴转者分别为56例(78.9%)和60例(92.3%).24周内,两组HBV DNA阴转率比较,x2=0.058,P＞0.05,差异无统计学意义.未抗病毒组HBV DNA≤106拷贝/ml 43例,HBV DNA＞106拷贝/ml 28例.24周内,HBV DNA≤106拷贝/ml组HBV DNA阴转41例(95.3%),HBV DNA＞106拷贝/ml组HBV DNA阴转21例(75.0%),HBV DNA＞106拷贝/ml的患者HBV阴转率低,两组比较,x2=0.024,P＜0.05,差异有统计学意义.未抗病毒组HBeAg阴性41例,HBeAg阳性30例,24周内,HBV DNA分别阴转36例(87.8%)和26例(86.7%),两组比较,x2=1,P＞0.05,差异无统计学意义.HBeAg阳性30例中,发生HBeAg阴转10例,有4例发生在4周内.未抗病毒组71例中,HBV DNA阴转62例,有5例HBV DNA反弹(发生在24～72周),均伴有ALT再次升高(47～140 U/L).结论 ALT＞800 U/L的CHB患者有自发清除HBV倾向,可以考虑暂时不抗病毒治疗.HBV DNA的阴转与HBeAg状态无关,但与HBV载量相关.     

慢性乙型肝炎患者12周自发性HBV DNA水平变化分析

目的 分析慢性乙型肝炎患者12周内自发性HBV DNA水平下降情况.方法 回顾性分析2003-2005年未接受抗病毒药物治疗的慢性乙型肝炎患者12周内自发性HBV DNA水平下降情况,并根据患者基线ALT、总胆红素(TBil)水平进行分组,分析基线ALT、TBil对自发性HBVDNA水平下降的影响.两组间计量资料比较采用t检验或Wilcoxon符号秩和检验;多组数据均值比较采用方差分析;组间率的比较采用x2检验.结果 共收集213例慢性乙型肝炎患者,男性174例,女性39例,年龄18～65(33.0±10.0)岁.其中慢性乙型肝炎轻～中度124例,慢性乙型肝炎重度89例;12周时失访19例(8.92%).所有患者HBV DNA基线均值为(6.66±1.03)log10拷贝/ml,12周时为(5.98±1.53)log10拷贝/ml(P<0.01).慢性乙型肝炎重度患者基线时HBVDNA水平均值低于慢性乙型肝炎轻～中度患者,分别为(6.45±0.99)log10拷贝/ml与(6.81±1.04)log10拷贝/ml(P<0.05);但两组12周时HBV DNA水平均值及HBV DNA水平下降值的差异均无统计学意义.12周时HBV DNA≤3 log10拷贝/ml患者的基线ALT及TBil值高于HBVDNA>3 log10拷贝/ml组,但差异均无统计学意义.12周时HBV DNA水平下降值≥2 log10拷贝/ml与<2 log10拷贝/ml两组患者的基线ALT、TBil水平相近(P>0.05).基线ALT水平≤5倍正常值上限(ULN)与>5×ULN两组患者12周时HBV DNA水平均值及HBV DNA水平下降值的差异均无统计学意义;两组患者12周时HBV DNA≤3 log10拷贝/ml、HBV DNA水平下降值≥2 log10拷贝/ml的比例,差异也无统计学意义(P>0.05).基线时ALT≤5 × ULN及TBil≤5×ULN组HBV DNA水平均值高于其他3组(P<0.05),但12周时各组HBV DNA水平均值、HBV DNA下降值比较,差异无统计学意义. 结论 慢性乙型肝炎患者12周内存在一定程度的自发性HBV DNA水平下降,但肝脏炎症损伤程度与患者12周内自发性HBV DNA水平下降程度无明显的相关性.     

慢性乙型肝炎抗病毒疗效与外周血单个核细胞内乙型肝炎病毒DNA水平的关系

目的 探讨慢性乙型肝炎(CHB)抗病毒疗效与达到停药标准时外周血单个核细胞(PBMC)内HBV DNA水平的关系.方法 入选90例经抗病毒治疗达到停药标准的CHB患者,其中应用IFN 44例,应用核苷类药物46例.所有患者均于停药时检测PBMC内HBV DNA,比较阴性组和阳性组治疗前血清HBV DNA水平与达到停药标准时PBMC内HBV DNA的关系,观察停药时PBMC内HBV DNA水平与复发的关系.计量资料采用t检验,计数资料采用X2检验.结果 90例CHB患者停药时,PBMC内HBV DNA阴性组67例,阳性组23例.CHB患者血清HBV DNA阳转率在PBMC内HBV DNA阴性组为13.4%(9/67例),显著低于阳性组的73.9%(17/23例),差异有统计学意义(X2=30.4873,P<0.01).PBMC内HBV DNA阴性组与阳性组在肝病复发ALT升高幅度(t=0.8729,P=0.3913)、停药后复发时间(t=1.9222,P=0.0665)均差异无统计学意义,而在血清HBV DNA反弹幅度则差异有统计学意义(t=2.7493,P=0.0112).5例患者获得HBsAg血清学转换,且均未检测到PBMC内HBV DNA,随访6～12个月无一例复发.PBMC内HBV DNA阳性组治疗前血清HBV DNA水平为(7.2±1.1)lg拷贝/mL,显著高于阴性组的(5.2±2.1)lg拷贝/mL(t=4.3557,P<0.01).结论 经抗病毒治疗达到停药标准的CHB患者,其停药时的PBMC内HBV DNA水平可能是预测抗病毒疗效持久性的重要因素之一.

Abstract：

Objective To explore the relationship between the antiviral effect and peripheral blood mononuclear cell (PBMC) hepatitis B virus (HBV) DNA when the patients reach the standard of withdrawal of antiviral therapy in chronic hepatitis B (CHB).Methods Ninety CHB patients treated with interferon(n=44) or nucleot (s) ide(n=46) who reached the standard of withdrawal of antiviral therapy were recruited.HBV DNA levels in PBMCs were tested at the end of treatment,and its relationship with serum HBV DNA level before treatment in PBMC HBV DNA positive group and negative group were compared.The correlation between HBV DNA in PBMCs at the end of treatment and relapse were explored.Measurement data were analyzed by student t test and enumeration data were analyzed by X2 test.Results Among 90 patients,67(74.4%) were PBMC HBV DNA negative at the end of treatment,and 23(25.6%) were positive.The serum HBV DNA positive conversion rate in PBMC HBV DNA negative patients was 13.4%,which were significantly lower than that in positive group (73.9%) (X2=30. 4873, P<0.01 ). There were no significant differences of alanine aminotransferase (ALT) levels when hepatitis flare (t=0. 8729, P=0. 3913) and relapse time (t=1. 9222, P=0. 0665) between PBMC HBV DNA negative group and positive group after withdrawal of therapy, while the serum HBV DNA rebound was greater in positive group than that in negative group (t=2. 7493, P=0. 0112). There were five patients who achieved hepatitis B surface antigen (HBsAg) seroconversion, whose PBMC HBV DNA were all undetectable, and none relapsed during follow-up for 6-12 months. The pretreatment HBV DNA as level in PBMC HBV DNA positive was (7.2±1.1) lg copy/mL, which was much higher than that in negative group[(5.2±2.1) lg copy/mL] (t=4. 3557, P<0.01). Conclusions In patients who reach the standard of drug withdrawal,PBMC HBV DNA at the end of treatment is an important predictor for durability of antiviral therapy in CHB.     

阿德福韦酯治疗HBeAg阳性慢性乙型肝炎病毒学应答的预测因素分析

目的 探讨阿德福韦酯(ADV)治疗HBeAg阳性慢性乙型肝炎(CHB)患者病毒学应答的预测因素.方法 对203例HBeAg阳性CHB患者采用ADV 10 mg/d治疗48周,PCR-限制性片段长度多态性检测TNF-α-238及TNF-α-308位点基因多态性,ELISA测定基线血清TNF-α水平,荧光定量PCR或HBV S基因直接测序法检测HBV基因型、亚型,Logistic回归分析影响ADV应答的因素.结果 203例患者ADV治疗24周和48周时HBV DNA转阴率、ALT复常率、HBeAg转阴率及转换率、联合应答率分别为31.5%(64/203)、59.1%(120/203)、15.8%(32/203)、8.9%(18/203)、13.3%(27/203)和58.6%(119/203)、78.3%(159/203)、29.6%(60/203)、16.7%(34/203)、25.6%(52/203).HBV基因型B、TNF-α-308G/A基因型、较高水平基线ALT及较低载量基线HBV DNA易于24周时HBV DNA转阴[OR=0.405,95%CI(0.191～0.859),P=0.019;OR=0.292,95%CI(0.132～0.643),P=0.002;OR=0.933,95%CI(0.989～0.997),P＜0.01;OR=2.089,95%CI(1.412～3.092),P＜0.01];24周HBV DNA高转阴率、较高水平基线ALT有利于48周时HBV DNA转阴[OR=0.029,95%CI(0.007～0.126),P＜0.01;OR=0.995,95%CI(0.991～0.999),P=0.016].结论 HBeAg阳性CHB患者ADV治疗24周病毒学应答的预测因素是HBV基因型、TNF-α-308基因型、基线ALT水平及HBV DNA载量;48周的预测因素是24周HBV DNA转阴率、基线ALT水平.

Abstract：

Objective To investigate the predictive factors of virological response in HBeAg-positive chronic hepatitis B (CHB)patients treated with adefovir dipivoxil (ADV).Methods A total of 203 HBeAg-positive CHB patients treated with ADV (Mingzheng)10 mg once daily for 48 weeks were recruited.The gene polymorphisms at positions-238 and-308 in tumor necrosis factor (TNF)-α promoter region were determined by the restriction fragment length polymorphism assay of products amplified using polymerase chain reaction (PCR-RFLP).The serum levels of TNF-a at baseline were measured by enzyme linked immunosorbent assay (ELISA).Hepatitis B virus (HBV)genotypes were tested by real-time fluorescent quantitative PCR and HBV subgenotypes were tested by HBV S gene sequencing.Factors related to ADV response were determined by Logistic regression analysis.Results The HBV DNA negative rate,alanine aminotransferase (ALT)normalization rate,HBeAg loss rate and seroconversion rate,and combined response rate at week 24 and 48 of treatment in 203 patients were 31.5% (64/203),59.1% (120/203),15.8% (32/203),8.9% (18/203),13.3% (27/203)and 58.6% (119/203),78.3% (159/203),29.6% (60/203),16.7% (34/203),25.6% (52/203),respectively.HBV DNA negative rate at week 24 was higher in patients with HBV genotype B,that was higher in patients with TNF-α-308G/A genotype,and that was higher in patients with higher baseline ALT level or lower baseline HBV DNA level [OR = 0.405,95 % CI (0.191 - 0.859),P =0.019;OR=0.292,95%CI(0.132-0.643),P=0.002;OR=0.933,95%CI(0.989-0.997),P＜0.01 ;OR=2.089,95%CI (1.412-3.092),P＜0.01].Meanwhile,HBV DNA negative rate at week 48 were higher in patients with higher HBV DNA negative rate at week 24 or higher baseline ALT level [OR=0.029,95%CI(0.007-0.126),P＜0.01;OR= 0.995,95%CI(0.991-0.999),P=0.016].Conclusions HBV genotype,TNF-α-308 genotype,baseline levels of ALT and HBV DNA are predictors of virological response at week 24 in HBeAg-positive CHB patients treated with ADV.And the HBV DNA negative rate at week 24 and baseline ALT level are predictors of virological response at week 48.     

替比夫定治疗HBeAg阳性慢性乙型肝炎的疗效及相关预测因子的Logistic回归分析

目的观察替比夫定（LdT）治疗HBeAg阳性慢性乙型肝炎（CHB）患者3年的疗效,应用Logistic回归探讨HBeAg血清学转换的预测因子。方法收集58例采用LdT治疗的HBeAg阳性CHB患者,分析其性别、年龄、基线ALT水平、基线HBV DNA载量、基线HBeAg和HBsAg滴度与治疗3年时ALT复常率、HBV DNA阴转率、HBeAg阴转率和HBeAg血清转换率的相关性;采用Logistic回归分析HBeAg血清转换的相关因素。结果治疗3年时ALT复常率为84.48%,HBV DNA阴转率为70.69%,HBeAg阴转率为50.00%,HBeAg血清转换率为43.10%。与ALT≤2倍正常值上限（2×ULN）相比,基线ALT〉5×ULN的患者HBeAg转换率显著增高（P〈0.05）;与HBeAg≤100（S/CO）组相比,基线HBeAg〉200 S/CO的患者HBeAg的阴转率和血清转换率均显著下降（P〈0.05）;与HBV DNA≤6 log拷贝/ml组相比,HBV DNA〉7 log拷贝/ml的患者HBV DNA转阴率、HBeAg转阴率和HBeAg转换率下降显著（P〈0.05）;患者性别、年龄及基线HBsAg滴度对以上疗效指标无影响（P〉0.05）。多因素Logistic回归分析发现仅基线HBeAg滴度低的患者更易出现HBeAg血清学转换。结论 LdT能有效恢复肝功能,抑制HBV复制和提高HBeAg血清转换;基线HBeAg滴度可预测LdT治疗HBeAg阳性CHB患者的HBeAg血清转换率。     

慢性乙型肝炎合并非酒精性脂肪性肝病患者抗病毒治疗的疗效探讨

目的对慢性乙型肝炎（CHB）合并非酒精性脂肪性肝病（NAFLD）患者抗病毒治疗疗效进行回顾性分析,探讨肝细胞脂肪变是否影响CHB患者抗病毒的疗效。方法收集2004年1月至2011年12月在本院进行抗病毒治疗的110例CHB患者,CHB合并NAFLD患者99例,采用荧光定量PCR法检测血清HBV DNA水平,用化学发光法检测血清乙型肝炎两对半定量,采用全自动生物化学分析仪检测肝功能。计数资料采用χ2检验,计量资料采用t检验。结果 （1）干扰素抗病毒治疗时,24周单纯CHB患者较合并脂肪肝患者生化学应答率更高（χ2=4.069,P=0.044）;48周HBV DNA阴转率明显优于后者（χ2=17.327,P=0.000）。对于HBeAg阳性患者,单纯CHB患者在24周的生化学应答率、24和48周时的HBV DNA阴转率、HBeAg阴转率均显著优于合并脂肪肝患者,两者比较差异具有统计学意义（P〈0.05）,48周时两组的生化学应答差异无统计学意义（P〉0.05）。（2）核苷和核苷酸类药物抗病毒治疗时,48周单纯CHB患者的生化学应答率较高（χ2=7.620,P=0.006）,24和48周的HBV DNA阴转率差异无统计学意义。对于HBeAg阳性患者,24周时合并脂肪肝组患者的HBeAg转阴率高于单纯CHB患者,48周时ALT/AST复常率低于单纯CHB患者,差异均具有统计学意义（P〈0.05）,24周ALT/AST复常率、HBV DNA转阴率、48周HBV DNA转阴率、HBeAg转阴率差异均无统计学意义（P〉0.05）。结论肝细胞脂肪变对CHB患者的抗病毒治疗效果存在一定的影响。     

血清HBsAg水平检测在干扰素联合阿德福韦酯治疗慢性乙型肝炎患者疗效评估中的意义

目的探讨血清HBs Ag水平检测在干扰素联合阿德福韦酯(ADV)治疗慢性乙型肝炎(CHB)患者疗效评估中的临床价值。方法应用化学发光法检测60例CHB患者在干扰素联合ADV治疗前和治疗过程中HBs Ag水平,以聚合酶链反应技术检测HBV DNA,并以连续监测法监测ALT水平并分析HBV DNA与HBs Ag的相关性。结果观察组30例CHB患者治疗前HBV DNA水平为(7.1±0.6)log10 IU/ml,HBs Ag水平为(3.5±0.4)log10 IU/ml。经干扰素联合ADV治疗3个月后HBs Ag下降,6个月后显著下降(P=0.000);治疗6个月后HBV DNA显著下降(P=0.000);治疗12个月后HBs Ag和HBV DNA均降至较低水平,分别为(2.1±0.3)log10 IU/ml和(2.7±0.3)log10 IU/ml;治疗3个月后ALT水平显著下降,治疗6个月后基本正常;治疗12个月无CHB患者产生耐药;患者HBs Ag水平与HBV DNA水平呈正相关。结论干扰素联合ADV治疗可显著降低慢性乙型肝炎患者血清ALT水平、提高HBe Ag低于检测下限的比率及HBV DNA低于检测下限的比率。定期检测HBs Ag浓度有助于评估ADV联合干扰素治疗CHB的疗效、及时发现耐药及是否需延长疗程。     

乙型肝炎病毒感染不同状态下血清病毒反转录酶区准种特点及其临床意义

目的 研究HBV感染不同状态下血清病毒反转录酶(RT)区准种特点和临床意义.方法 50例未接受抗病毒治疗的HBV感染者分为慢性HBV携带者(ASC)组10例、慢性乙型肝炎(CHB)组30例和乙型肝炎肝硬化(LC)组10例.收集患者外周血血清,抽提HBV DNA,PCR扩增RT区基因组后克隆、测序.每例患者获得15～30个序列,进行HBV RT区的准种异质性分析及相关基因突变统计等生物信息学分析.多均数比较采用方差分析,中位数比较采用非参数检验分析,非计量资料比较采用x2检验.结果 总共测序1221个克隆,其中ASC组152个,CHB组780个,LC组289个.3组间基因型构成差异无统计学意义.准种复杂度为LC组＞CHB组＞ASC组,3组间差异有统计学意义(F=33.400,P＜0.05).准种离散度为LC组＞CHB组＞ASC组,LC组与CHB组及ASC组均差异有统计学意义(F=18.070,P＜0.05),CHB组与ASC组间差异无统计学意义.结论 慢性HBV感染过程中,免疫清除期比免疫耐受期具有更宽的HBV变异谱系,随着病程的延长和病情加重,HBV的准种趋向复杂.     

丙氨酸转氨酶正常的慢性乙型肝炎病毒感染者的肝脏病理学分析

目的 了解ALT正常的慢性HBV感染者的肝脏病理学改变及其影响因素.方法 观察632例ALT正常的慢性HBV感染者,采用超声定位穿刺取肝组织,行HE染色、纤维Masson染色,HBsAg和HBcAg免疫组织化学染色,观察Knodell坏死炎症评分和Ishak纤维化评分,并分析它们与年龄、ALT水平、血清HBV DNA载量、HBsAg和HBcAg肝组织表达的关系.两均数比较采用t检验,多均数比较采用单因素方差分析及q检验,计数资料采用x2检验.结果 632例ALT正常的HBV感染者中,中度炎症坏死167例,占26.4%,重度炎症坏死26例,占4.1%,中度纤维化217例,占34.3%,重度纤维化(肝硬化)52例,占8.2%.Knodell坏死炎症评分和Ishak纤维化评分在高ALT层次组比低ALT层次组高,在女性高ALT层次组比男性高ALT层次组高,在年龄＞40岁组比年龄≤20岁组高(q=19.63,P＜0.05).肝组织损伤程度在HBV DNA载量≤5×105拷贝/L组明显轻于HBV DNA 5×105～1×107拷贝/L、1×107～1×109拷贝/L和＞1×109拷贝/L组(Knodell评分,q=3.87、2.87、6.34;Ishak评分,q=2.64、2.64、5.54;均P＜0.05),在不同HBV DNA载量复制组之间差异无统计学意义(F=1.35,P＞0.05).HBsAg(F=1.65、0.73,均P＞0.05)和HBcAg(F=0.17、1.29,均P＞0.05)肝组织表达与Knodell坏死炎症评分和Ishak纤维化评分差异均无统计学意义.结论 可检测到HBV DNA的ALT持续正常的慢性HBV感染患者应考虑进行肝组织活检,特别是年龄＞40岁且ALT在(0.75～1.00)×正常值上限者.     

慢性乙型肝炎HBsAg及HBsAg/HBV DNA比值与组织病理炎症活动度的相关性

目的 研究HBeAg阳性慢性乙型肝炎(CHB)基线血清HBsAg、HBsAg/HBVDNA比值与肝组织病理炎症活动度的相关性. 方法 回顾性分析153例HBeAg阳性CHB患者基线HBsAg、HBsAg/HBV DNA比值与肝组织病理炎症活动度的相关性.采用Taqman荧光定量PCR法检测血清HBV DNA水平,定量检测血清HBsAg滴度. 结果 HBsAg (log10IU/ml)与HBV DNA (log10IU/ml)、HBsAg/HBV DNA比值进行相关性分析,相关系数r分别为0.642、0.57,P值均＜ 0.0001,均呈显著性正相关;HBsAg和HBsAg/HBV DNA比值与炎症活动度进行相关性分析,相关系数r分别为-0.389、-0.307,P值均＜0.0001,二者与炎症活动度均呈负相关;而ALT (log10U/L)与炎症活动度呈正相关(r=0.480,P＜0.0001).肝组织炎症活动度中度及以上患者的血清HBsAg及HBsAg/HBV DNA比值均显著低于轻度及以下患者,组间差异具有统计学意义(P均＜ 0.01).HBsAg、HBsAg/HBV DNA比值及ALT在组织炎症活动度最优截断点的受试者工作曲线下面积分别为0.700、0.672、0.713;当机会曲线下面积等于0.5时,其显著性水平均有统计学意义(P均＜0.001).HBsAg诊断组织炎症活动度的灵敏度为76.92％高于ALT的4.36％;HBsAg/HBV DNA比值的特异度为81.33％高于ALT的64.00％,ALT的约登指数均高于HBsAg及HBsAg/HBV DNA比值.当HBsAg与ALT并联时,其灵敏度高达94.08％;串联时,其特异度可高达85.60％. 结论 HBeAg抗原阳性CHB患者HBsAg、HBsAg/HBV DNA比值及ALT均可作为肝组织炎症程度的判断指标,HBsAg与ALT并联或串联诊断时具有更高的灵敏度和特异度.     

乙型肝炎病毒和丙型肝炎病毒不同模式重叠感染患者临床特征分析

目的 了解HBV和HCV不同模式重叠感染患者临床特征的差异.方法 回顾分析中山大学附属第三医院1999年5月至2010年5月HCV和HBV重叠感染患者186例.统计分析不同病毒学模式重叠感染患者的人口学、流行病学、实验室检查和病理学表现,数据处理采用t检验和x2检验等.结果 186例HBV和HCV重叠感染患者中,HBV...     

恩替卡韦治疗后慢性乙型肝炎患者外周血T淋巴细胞表面程序性死亡受体1表达与HBeAg血清学转换的关系

目的 动态观察慢性乙型肝炎患者恩替卡韦抗病毒治疗后不同时期外周血T淋巴细胞(简称T细胞)表面程序性死亡受体1(PD-1)表达的变化,并探讨其与HBeAg血清学转换间的关系.方法 对20例HBeAg阳性慢性乙型肝炎患者予以恩替卡韦抗病毒治疗并随访51周,根据HBeAg是否发生血清学转换分为:HBeAg未转换组(14例),HBeAg转换组(6例).分别于治疗前(基线,T0)、治疗2～4周(T1)、治疗5～10周(T2)、治疗11～20周(T3)、治疗21～30周(T4)、治疗31～51周(T5)收集外周血,流式细胞术检测CD4+、CD8+T细胞表面PD-1的表达水平,实时荧光定量PCR检测血清HBV DNA载量,同时检测血清ALT水平.正态分布资料采用独立样本t检验,非正态分布者采用Mann-Whitney U检验比较组间差异,相关性分析采用Pearson相关分析.结果 治疗前两组患者血清HBV DNA载量分别为(7.54±0.67)log10拷贝/ml、(7.30±0.79)log10拷贝/ml(P＞0.05),ALT水平为(187.26±184.15)U/L、(272.17±215.07)U/L(P＞0.05),外周血CD4+T细胞表面PD-1表达水平为6.04%±3.71%6.77%±2.88%(P＞0.05),CD8+T细胞表面PD-1表达水平为6.39%±3.33%、8.88%±2.84%(P＞0.05).恩替卡韦抗病毒治疗后两组患者血清HBV DNA载量、ALT水平的下降与CD4+、CD8+T细胞表面PD-1表达的下调呈显著正相关(r=0.212,P=0.05;r=0.377,P＜0.01;r=0.279,P＜0.05;r=0.347,P＜0.01).在相同的随访时间段内,HBeAg转换组血清HBV DNA载量、ALT水平及外周血CD4+、CD8+T细胞表面PD-1表达的下降率均高于HBeAg未转换组,且两组间△ T0～T1、△T0～T2期HBV DNA的下降率及△T0～T2、△T0～T3期CD8+T细胞表面PD-1表达的下降率差异有统计学意义(分别为49.9%对比37.3%,56.7%对比47.4%,70.1%对比-4.2%,66.9%对比24.5%,P值均＜0.05).结论 HBeAg阳性慢性乙型肝炎患者经恩替卡韦抗病毒治疗后,外周血CD8+T细胞表面PD-1表达的快速下调与血清HBV DNA相似,可作为预测后期HBeAg血清学转换的指标之一.

Abstract：

Objective To observe longitudinally the expression of Programmed death 1 (PD-1) on peripheral blood T cells in chronic hepatitis B patients underwent antiviral treatment with entecavir (ETV)and to explore the relationship between PD-1 expression and HBeAg seroconversion.Methods Twenty HBeAg positive patients underwent antiviral treatment with ETV were followed up for 51 weels.14 patients remained HBeAg positive and 6 patients achieved HBeAg seroconversion.Peripheral blood was collected at six time points:T0:baseline,T1:2-4week;T2:5-10week;T3:11-20week;T4:21-30week:T5:31-51week.PD-1 expressions on T cells were assessed by flow cytometry.Serum HBV DNA loads were determined by real-time fluorescent quanttative polymerase chain reaction (PCR) and serum ALT levels were examined at the same time.Results At baseline,serum HBV DNA load of patients without HBeAg seroconversion and with HBeAg seroconversion were (7.54 ± 0.67) log10 copies/ml and (7.30 ± 0.79) log10 copies/ml(P ＞ 0.05),the ALT levels were (187.26 ± 184.15) U/L and (272.17 ± 215.07) U/L (P ＞ 0.05),PD-1 exprissions on CD4+ T cells were 6.04% ± 3.71% and 6.77% ± 2.88% (P ＞ 0.05),PD-1 exprissions on CD8+ T cells were 6.39% ± 3.33% and 8.88% ± 2.84% (P ＞ 0.05).After ETV treatment,serum HBV DNA loads and ALT levels both decreased gradually,which was positively correlated with PD-1 expressions on CD4+ and CD8+ T cells (r = 0.212,P = 0.05;r = 0.377,P ＜ 0.01;r = 0.279,P ＜ 0.05;r = 0.347,P ＜ 0.01 ).During the same monitoring period,the HBV DNA loads,ALT levels and PD-1 expressions on T cells of the patients with HBeAg seroconversion decreased significantly as compared with the patients without HBeAg seroconversion.Besides,the decrease of HBV DNA loads during period △ T0-T1 and △ T0-T2 and PD-1 expressions on CD8+ T cells during period △ T0-T2 and △ T0-T3 were significantly different between these two kinds of patients (49.9% vs 37.3%,P ＜ 0.05;56.7% vs 47.4%,P ＜ 0.05;70.1% vs -4.2%,P ＜ 0.05;66.9% vs 24.5%,P ＜ 0.05).Conclusion The rapid decrease of PD-1 expression on peripheral CD8+ T cells after antiviral treatment with ETV is positvely correlated with the decrease of serum HBV DNA loads and may be used as a predictive index for HBeAg seroconversion in HBeAg positive patients.     

乙型肝炎患者血清中乙型肝炎病毒共价闭合环状DNA的检测

目的 检测不同肝脏病变程度的乙型肝炎患者外周血中的HBV共价闭合环状DNA(HBV cccDNA),评价其相关的影响因素和临床意义.方法 共观察57例乙型肝炎患者,其中轻度慢性乙型肝炎患者26例,重型乙型肝炎患者31例.患者入院后行PT、肝功能、肝炎病毒血清标志物等常规检测,并进行总HBV DNA和HBV cccDNA检测.Logistic逐步回归分析影响血清HBVcecDNA检出率的相关因素.结果重型乙型肝炎组13份血清标本HBV cccDNA为阳性,轻度慢性乙型肝炎组仅1份血清标本HBV cccDNA为阳性,血清HBV cccDNA的含量为1.25×103～4.88×104拷贝/mL,两组患者血清HBV cccDNA检出率差异有统计学意义(P=0.0014).血清HBV cccDNA检测诊断重型乙型肝炎患者的灵敏度和特异度分别为41.94%和96.15%.Logistic逐步回归分析显示,血清HBV ccvDNA的检出率与PT有关(X<'2>=7.2192,P=0.0072),而与患者的年龄、性别、血清总HBV DNA、TBil和ALT水平无相关性.结论 部分乙型肝炎患者特别是重型肝炎患者外周血中可以检测出HBV cccDNA,血清HBV cccDNA的检出可作为支持重型乙型肝炎诊断的指标之一.     

阿德福韦酯联合安络化纤丸治疗慢性乙型肝炎的疗效

目的 观察阿德福韦酯(ADV)联合安络化纤丸治疗慢性乙型肝炎患者的临床疗效.方法 治疗组36例采用ADV联合安络化纤丸治疗,对照组36例单用ADV治疗,治疗48周后,观察治疗前后的肝功能、肝纤维化指标、HBV DNA及肝组织学.统计学分析采用t检验或x2检验.结果 治疗48周后,治疗组和对照组患者在肝功能、血清纤维化标志物及肝脏组织学均有改善,治疗后两组患者在ALT、AST、总胆红素、白蛋白、肝组织炎症活动度积分和HBV DNA阴转率方面比较,差异无统计学意义.治疗组和对照组患者治疗后血清肝纤维化标志物透明质酸(HA)值分别为(101.58±30.11)μg/L、(182.25±117.59)μg/L;层黏连蛋白(LN)值分别为(147.89±41.72)μg/L、(181.50±56.96)μg/L;Ⅳ型胶原蛋白(CⅣ)值分别为(38.75±9.50)μg/L、(74.92±31.14)μg/L,治疗组患者的HA、LN、CIV值与对照组比较,t值分别为3.987、2.856、6.666,P值均＜0.05,差异均有统计学意义.治疗组治疗前后肝组织纤维化积分分别为12.28±3.16、10.61±2.37,治疗前后比较,t=2.532,P＜0.05,差异有统计学意义;对照组治疗前后肝组织纤维化积分分别为12.17±3.01、11.36±2.93,治疗前后比较,t=1.151,P＞0.05,差异无统计学意义.结论 ADV联合安络化纤丸治疗慢性乙型肝炎患者具有良好的抗肝纤维化作用,患者获得明显肝组织学改善.     

慢性乙型肝炎患者重叠戊型肝炎病毒感染的临床与病理学研究

目的：观察重叠戊型肝炎病毒（HEV）感染对慢性乙型肝炎（CHB）肝脏损害及HBV复制的影响。方法：应用ELISA法对122例CHB患者血清进行了抗－HEV IgkM,IgG检测,同时应用肝穿刺活检、荧光定量PCR及免疫组化等技术对重叠与未重叠HEV感染者分别进行了ALT、总胆红素（TBil)、凝血酶原活动度（PTA）、白蛋白／球蛋白（A／G）、电泳γ球蛋白（γ－EP）水平、肝脏病理学、血清HBeAg及肝组织HBcAg阳性率、血清及肝组织中HBV DNA含量对比。具有可比性的重叠（7例）与未重叠HEV感染者（14例）1年后做第2次肝穿活检并做病理学比较;HBeAg阴性重叠HEV感染者8例做HEV感染急性期、恢复期血清HBeAg定性、HBV DNA含量对比。结果：重叠HEV感染者21例（17．2％）。重叠HEV感染者较未重叠感染者ALT、TBil增高,PTA降低（P＜0．05）,但A／G、γ－EP水平未见显著差别（P＞0．05）;血清HBeAg及肝组织HBcAg阳性率、血清及肝组织HBV DNA含量低（P＜0．05）;肝组织炎症活动度重（P＜0．05）,但纤维化程度未见明显差别（P＞0．05）。两组患者1年前肝组织炎症活动度及纤维化程度无显著差别,1年后仍无显著差别（P＞0．05）。HEV感染恢复期血清HBeAg阳性率、HBV DNA含量高于急性期（P＜0．05）。结论：重叠HEV感染可加重CHB肝组织炎症活动度;对HBV复制具有短暂抑制作用。     

Prognostic Values of Fluctuations in Serum Levels of Alanine Transaminase in Inactive Carrier State of HBV Infection

Background:

Current guidelines introduce periodic monitoring of serum alanine transaminase (ALT) as the first-line modality in follow-up patients, with a hepatitis B virus (HBV) inactive carrier state.

Objectives:

This study aimed to determine the incidence rate and patterns of ALT fluctuations and prognostic values for the development of chronic HBV e antigen (HBeAg)-negative hepatitis B (CHB), HBV surface antigen (HBsAg) seroclearance, and liver-related complications.

Patients and Methods:

Treatment-naïve patients with a chronic HBV infection, HBeAg(-)/HBeAb(+), normal ALT levels, and HBV DNA < 2000 IU/mL, were followed-up every 6-12 months by assessing serum ALT levels. Serum HBV DNA was measured in cases of elevated ALT levels.

Results:

A total of 399 patients were followed-up for 8.9 years; ALT > upper limit of normal (ULN, i.e. 40 IU/L) was detected in 103 (25.8%) patients, with an annual incidence rate of 2.9%. ALT elevation was associated with; male gender, age, and higher serum ALT levels at study entry. Among the cases of ALT elevations, 16 (15.5%) patients had ALT levels > 2 × ULN. There were 38 (36.9%) patients who had ALT levels that remained > ULN over six months, and 21 (20.4%) patients experienced at least two episodes of ALT elevations. In 15 (14.6%) patients, elevated ALT levels were associated with increased HBV replication (i.e. HBV DNA > 2 000 IU/mL) and these were considered as CHB. However, elevation of ALT levels, even in the absence of HBV replication, increased the risk for the development of CHB up to 8-fold in prospective follow-ups. HBsAg seroclearance, cirrhosis, and hepatocellular carcinoma were detected in 43 (10.8%), 4 (1%), and 1 (0.25%) patients, respectively.

Conclusions:

Fluctuations in serum ALT levels may change the prognosis of a HBV inactive carrier state.