乙肝患者血清的HBV-DNA含量与ALT水平的关系研究

目的:探讨和研究乙型肝炎患者乙型肝炎病毒(HBV)DNA复制水平与丙氨酸氨基转移酶(ALT)水平之间的关系,为临床中乙肝患者的诊治提供参考。方法:对240例慢性乙型肝炎患者采用荧光标记定量PCR方法测定血清HBV-DNA含量,采用全自动生化分析仪测定血清ALT水平,比较和分析HBV-DNA含量与ALT水平之间的关系。结果:慢性乙肝轻、中、重度患者的HBV-DNA含量三组之间差异有统计学意义(p0.01),肝脏的损害程度与HBV-DNA含量之间具有一定的关联,等级相关系数为0.162(P=0.012);ALT水平也与HBV-DNA载量之间存在关联,等级相关系数为0.371(P0.0001)。结论:肝损伤程度与HBV-DNA含量有显著相关性;同时血清ALT水平与HBV-DNA含量呈正相关。检测血清中的HBV-DNA含量和ALT水平为指导乙肝患者HBV感染、复制、传染性的判断、治疗方案的选择和疗效评定提供有一定的依据。     

乙型肝炎血清标志模式与病毒载量的关系及意义

为探讨乙型肝炎 (以下简称乙肝 )血清标志模式与病毒载量的关系及临床意义 ,作者选择符合 2 0 0 0年《全国病毒性肝炎诊断标准》的慢性肝炎血清 1343份 ,分别用ELISA法、PCR ELISA法检测HB血清标志、HBV -DNA和 1896位点变异株。结果显示 :HBsAg阳性血清 10 97份 (81.6 8%、)HBsAg阴性血清 2 4 6份 (81.31% )。在HBsAg阳性血清中 ,HBsAg、HBeAg、抗 HBc(1 3 5 )阳性组 4 0 4份 (30 .l% ) ,HBV -DNA阳性 347份 (85 .89% ) ,DNA阳性值呈递增趋势 (10 4～ 10 6拷贝 /ml,各占 8.6 5 %、33.71%、5 3.6 1% ) ;而HBsAg、抗 HBe阳性组血清 6 0 6份 (45 .12 % ) ,DNA阳性值 10 5拷贝 /ml,占优势 (6 4 .18% )。在HBsAg阴性血清中 ,抗 HBs、抗 HBe、抗 HBc(2 4 5 )阳性组 2 32份 (17.2 7% ) ,DNA阳性占 7.32 % ,DNA阳性值递减由 10 4～ 10 6拷贝 /ml,各占 5 2 .9%、4 1.l%、5 .9%。结论 ,各血清标志模式中的病毒载量为HBsAg阳性组 >HBsAg阴性组 ,阳性组 1 3 5 >1 4 5 >l 5 >2 4 5。但 1 3 5阳性组中 14 %在界值以下 ,1 4 5阳性组中 1896位点自然变异达 78.6 % ,2 4 5阳性组中仍存在DNA+ 血清 ,以上提示在临床判定和治疗时要慎重对待(注 :l=HBsAg,2 =抗 HBs,3=HBeAg ,4 =抗 HBe ,5 =抗 HBc)     

不同临床类型乙型病毒感染者外周血T 细胞亚群与血清HBV DNA 载量 及HBeAg 滴度相关性分析

目的:探讨慢性乙型肝炎病毒(HBV)感染患者外周血T细胞亚群与血清HBV DNA载量及HbeAg滴度的关系。方法:选取103名HBV感染患者和20名健康者为研究对象。流式细胞术检测外周血T细胞亚群,聚合酶链式反应及酶免疫分析法分别检测血清HBV DNA载量及HbeAg滴度。结果:慢性乙型肝炎患者和慢性HBV携带者外周血CD3+T、CD4+T淋巴细胞亚群百分数低于健康对照组,结果有统计学意义(P<0.05或0.01;而CD8+T细胞亚群则呈现相反趋势,结果亦有统计学意义(P<0.05或0.01)。HBeAg阴性组中,HBVDNA水平与CD8+T细胞亚群百分数呈正相关(r=0.567,P<0.01),与CD4+/CD8+T细胞亚群百分数比值呈负相关(r=-0.601,P<0.01),而与CD3+T、CD4+T细胞亚群百分数无相关性。HBeAg阳性组中,HBV DNA水平及HbeAg滴度与CD3+T、CD4+T、CD8+T细胞百分数及CD4+/CD8+T细胞百分数均无相关性(P>0.05)。结论:不同临床类型的慢性乙型肝炎病毒感染患者外周血T细胞亚群存在不同程度细胞免疫功能降低和细胞免疫调节异常。HbeAg阴性的HBV感染患者,其血清HBV DNA水平与外周血T淋巴细胞免疫存在相关性。     

乙型肝炎病毒前S1抗原与HBV血清标志物同时检测及临床意义

目的：了解前S1抗原与HBV血清标志物的关系。方法：采用ELSIA方法同时检测2101例乙型肝炎病毒感染者的前S1抗原和HBV血清标志物。结果：29例HBsAg( )、HBeAg( )标本中,有28例前S1抗原阳性,阳性率96．55％;784例HBsAg( )、HBeAg( )、抗HBc( )标本中,有679例前S1抗原阳性,阳性率86．6l％;276例HBsAg( )、抗HBc( )标本中,有197例前S1抗原阳性,阳性率71．38％;1012例HBsAg( )、抗Hbe( )、抗HBc( )标本中,有468例前S1抗原阳性,阳性率46．25％。结论：前S1抗原作为病毒复制的指标与HBeAg具有很好的一致性,又具有其独立的检测价值,可弥补HBV血清标志物检测的不足。     

乙型肝炎病毒前S1抗原与HBV血清标志物同时检测及临床意义

目的 :了解前S1抗原与HBV血清标志物的关系。方法 :采用ELSIA方法同时检测 2 1 0 1例乙型肝炎病毒感染者的前S1抗原和HBV血清标志物。结果 :2 9例HBsAg( )、HBeAg( )标本中 ,有 2 8例前S1抗原阳性 ,阳性率 96.5 5 % ;784例HBsAg( )、HBeAg( )、抗HBc( )标本中 ,有 679例前S1抗原阳性 ,阳性率 86.61 % ;2 76例HBsAg( )、抗HBc( )标本中 ,有 1 97例前S1抗原阳性 ,阳性率 71 .38% ;1 0 1 2例HBsAg( )、抗Hbe( )、抗HBc( )标本中 ,有 468例前S1抗原阳性 ,阳性率 46.2 5 %。结论 :前S1抗原作为病毒复制的指标与HBeAg具有很好的一致性 ,又具有其独立的检测价值 ,可弥补HBV血清标志物检测的不足。     

不同临床类型乙型病毒感染者外周血T细胞亚群与血清HBVDNA载量及HBeAg滴度相关性分析

目的：探讨慢性乙型肝炎病毒（HBV）感染患者外周血T细胞亚群与血清HBVDNA载量及HbeAg滴度的关系。方法：选取103名HBV感染患者和20名健康者为研究对象。流式细胞术检测外周血T细胞亚群,聚合酶链式反应及酶免疫分析法分别检测血清HBVDNA载量及HbeAg滴度。结果：慢性乙型肝炎患者和慢性HBV携带者外周血CD3可、CD4T淋巴细胞亚群百分数低于健康对照组,结果有统计学意义（P〈0．05或0．01;而CD8＋T细胞亚群则呈现相反趋势,结果亦有统计学意义（P〈0．05或0．01）。HBeAg阴性组中,HBVDNA水平与CD8T细胞亚群百分数呈正相关（r=0．567,P〈0．01）,与CD47CD8＋T细胞亚群百分数比值呈负相关（r=-0．601,P〈0．01）,而与CD3＋T、CD4＋T细胞亚群百分数无相关性。HBeAg阳性组中,HBVDNA水平及HbeAg滴度与cD3＋1r、cD41、CD8叮细胞百分数及CD47CD8＋T细胞百分数均无相关性（P〉0．05）。结论：不同临床类型的慢性乙型肝炎病毒感染患者外周血T细胞亚群存在不同程度细胞免疫功能降低和细胞免疫调节异常。HbeAg阴性的HBV感染患者,其血清HBVDNA水平与外周血T淋巴细胞免疫存在相关性。     

HBV感染者外周血PBMC中TLR4 mRNA与病毒载量的相关性研究

目的探讨乙型肝炎病毒感染者外周血PBMC中TLR mRNA与乙肝病毒复制的相关性。方法采用逆转录PCR检测外周血单个核细胞（PBMC）中TLR4 mRNA的含量,实时荧光定量Real Time PCR的方法检测HBV DNA,进行相关性分析。结果不同病毒载量组（〈1×103copies/μg DNA,1×103copies/μg DNA〈且〈1×105copies/μg DNA,〉1×105copies/μg DNA）TLR4 mRNA水平差异具有显著性（P〈0.01）,HBV病毒载量的对数值与TLR4 mRNA的含量存在负相关（r=-0.537,P〈0.01）。TLR4 mRNA的相对表达量与患者的ALT、AST呈正相关（r=0.608、r=0.659,P〈0.01）。结论HBV在患者体内复制活跃、病毒载量增高与外周血单个核细胞TLR4mRNA的表达下调有关。     

不同方法检测血清HBV DNA的比较研究

采用夹心斑点杂交法、PCR-EB定性及Amp-lisensor定量法对HBsAg与抗-HBs同时阳性、HBsAg阳性、抗-HBs阳性和HBsAg与抗-HBs均阴性的乙型肝炎病毒(HBV)感染者检测血清HBV DNA.结果三种方法的阳性率分别为29.1%、47.7%和80.2%;各组HBV DNA含量分别是106.13±1.86、106.08±1.82、104.12±1.40和105.07±1.21(拷贝/ml).结论是Amplisensor法检测HBV DNA具有更高的敏感性;抗-HBs阳性的HBV感染者血清中仍可检出HBV DNA.     

乙肝病毒与原发性肝癌的相关风险研究

为了解慢性乙型肝炎病毒感染与原发性肝癌的关系,本文采用回顾性研究方法对 328 例原发性肝癌病人与同期收治的 340 例非肝癌的其他消化道肿瘤病人的乙型肝炎病毒感染血清标志物(HBV M)及肝功能检测结果进行对比分析。结果显示肝癌组乙肝表面抗原(HBsAg)阳性率(63.11%)显著高于非肝癌组(消化道其他肿瘤对照组)(11.47%)。肝癌组慢性乙型肝炎病毒感染“HBsAg、抗-HBe 和抗-HBc 三者均表达为阳性者”(37.2%)显著高于“HBsAg、HBeAg 和抗-HBc 三者均表达为阳性者”(6.4%)。肝功能检测结果,“HBsAg、HBeAg 和抗-HBc三者均表达为阳性组”与“HBsAg、HBeAg 和抗-HBc 三者均表达为阳性组”比较无显著性差异(P>0.05),而肝癌组与非肝癌组比较,肝癌组肝损害显著高于非肝癌组(P<0.01)。表明慢性乙型肝炎病毒感染在原发性肝癌病因学中起着十分重要的作用,“HBsAg、抗-HBe 和抗-HBc 三者均表达为阳性者”是原发性肝癌的高危人群。     

ERCP术与腹腔镜手术治疗胆总管结石的疗效对比及对血清AST、ALT、ALP的影响

目的:探讨内镜逆行胰胆管造影术(ERCP)与腹腔镜手术治疗胆总管结石的疗效对比及对血清门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、碱性磷酸酶(ALP)的影响。方法:选择2017年4月到2018年4月我院接诊的胆总管结石患者50例作为研究对象,以随机数表法分为观察组(n=26)和对照组(n=24),对照组使用腹腔镜手术治疗,观察组采用ERCP取石术治疗。比较两组手术时间、术中出血量、术后下床时间、住院时间、血清AST、ALT、ALP水平变化情况、术后胃肠道恢复时间及术后不良反应发生情况。结果:观察组患者手术时间高于对照组,(P0.05),术中出血量、术后下床时间及住院时间均显著低于对照组(P0.05);手术前,两组患者血清AST、ALT、ALP水平无明显差异;治疗后,两组患者血清AST、ALT、ALP水平均明显上升(P0.05),且观察组患者血清AST、ALT、ALP水平显著低于对照组(P0.05);观察组患者肠鸣音恢复时间、排气时间、排便时间均明显低于对照组(P0.05);治疗期间,观察组患者不良反应总发生率为7.69%,显著低于对照组的29.17%(P0.05)。结论:在胆总管结石患者中应用ERCP取石术效果显著,对患者血清血清AST、ALT、ALP影响较小,值得推广与运用。     

慢性乙型肝炎患者血清HBV基因分型

为了解长春市慢性乙型肝炎患者血清中的乙型肝炎病毒(HBV)基因型情况及其与临床特点的相关性,应用型特异性引物进行巢式PCR方法对长春市69例慢性乙型肝炎患者血清HBV进行基因分型检测。在69例血清标本中,B型10例(占14.5%);C型41例(占59.4%);B C混合型8例(占11.6%);未分型的患者共10例(占14.5%)。C基因型患者的HBV-DNA定量、HBeAg阳性率明显高于B基因型患者(HBV-DNA:P<0.01;HBeAg:χ2=3.98,P<0.05),C基因型患者肝功检查指标谷丙转氨酶(ALT)和总胆红素(TB IL)均较B基因型患者高(P<0.01)。长春地区存在HBV B基因型、C基因型、B C混合基因型及未分型,C基因型为优势基因,引起的肝脏活动性炎症较B基因型明显。     

Effect of Vitamin E on Serum Aminotransferase and Thioredoxin Levels in Patients with Viral Hepatitis C

Objectives: Oxidative stress induces cellular responses such as cell death, gene activation and cell proliferation, in the liver. Vitamin E (Vit. E) has been found to protect the liver against oxidative stress in animal experiments. Thioredoxin (TRX) is a stress inducible, multifunctional protein, secreted during oxidative stress. This study evaluated effects of Vit. E on serum TRX and aminotransferase levels in hepatitis C virus (HCV) patients, partly non-responsive to initial interferon (IFN), with higher than average level of serum alanine aminotransferase (ALT) after receiving anti-inflammatory drug treatment. Methods: Seventeen HCV patients (male=3; female=14) of age 62±7.65 years receiving anti-inflammatory drug therapy, at least 6 months prior to Vit. E administration, were given d-α´-tocopherol 500?mg/day, orally, for a period of 3 months. ALT, aspartate aminotransferase (AST), TRX and Vit. E were measured at 0, 1, 2 and 3 months and 1 month after end of treatment. As controls, the same patients biochemical data, 3 months from the start of therapy were used. Patients were divided into three categories: total patients “T”, low ALT group “L” (ALT<70?IU/l) and high ALT group “H”(ALT>70?IU/l), respectively.Results: The ALT level was lowered, significantly in group H, in the 1st, 2nd, 3rd and 1-month post therapy, compared to the initial value. But group L showed little or no change in ALT. Post Vit. E therapy, in groups T and H, the TRX level was elevated but remained below initial levels, whereas in group L, TRX level remained significantly lower than the pretreatment value. Groups T and L, showed significant reduction (p<0.05) in serum TRX levels in the 2nd and 3rd month. Group H showed a tendency towards TRX reduction, but not significantly. Serum Vit. E levels increased significantly (p<0.0001) from the 1st to 3rd month in all three T, H and L groups. Conclusion: Oxidative stress induced liver damage is reduced by Vit. E in patients with viral hepatitis C, particularly those with initial ALT levels >70?IU/l. Vit. E treatment causes reduction of oxidative stress markers as TRX and ALT in sera. Therefore, Vit. E can act as a supportive therapy to combat liver damage caused by oxidative stress, in such patients with continuously high levels of ALT even after anti-viral and anti-inflammatory drug therapy.     

Generation of Hepatitis B virus PreS2-S antigen in Hansenula polymorpha

Despite the long availability of a traditional prophylactic vaccine containing the HBV surface antigen(HBsA g) and aluminum adjuvant, nearly 10% of the population remains unable to generate an effective immune response. Previous studies have indicated that hepatitis B virus(HBV) PreS 2-S is abundant in T/B cell epitopes, which induces a stronger immune response than HBsA g, particularly in terms of cytotoxic T lymphocyte(CTL) reaction. In the current study, the HBV PreS 2-S gene encoding an extra26 amino acids(PreS 2 C-terminus) located at the N-terminus of HBsA g was cloned into the pV CH1300 expression vector. Pre S2-S expressed in the methylotrophic yeast, Hansenula polymorpha, was produced at a yield of up to 250 mg/L. Subsequent purification steps involved hydrophobic adsorption to colloidal silica, ion-exchange chromatography and density ultracentrifugation. The final product was obtained with a total yield of ~15% and purity of ~99%. In keeping with previous studies, ~22 nm viruslike particles were detected using electron microscopy. The generated PreS 2-S antigen will be further studied for efficacy and safty in animals.     

乙肝病毒感染对细胞基本自噬的影响

【目的】慢性乙肝病毒(Hepatitis B virus,HBV)感染在肝硬化和肝癌的发生过程中起着重要的作用,通过研究HBV感染对细胞基本自噬的影响,为HBV感染诱发肝癌以及HBV的免疫逃逸机理研究提供新的思路。【方法】本研究利用乙肝病毒表达质粒瞬时或稳定转染不同肝细胞,通过计数绿色荧光蛋白(greenfluorescent protein,GFP)聚集数目检测自噬小体形成,western blot检测LC3(microtubule-associated proteinlight chain 3,微管相关蛋白质轻链3)脂酰化和p62的降解,通过构建HBV B型和C型X蛋白(HBx)的表达质粒并瞬时转染肝癌细胞和正常肝细胞,对不同基因型X蛋白对细胞自噬的影响进行了分析。【结果】乙肝病毒感染后促进了LC3的脂酰化和p62的降解,增加了自噬小体的形成,增强了细胞的基本自噬。进一步研究发现,HBV感染增强的细胞基本自噬水平由HBx所引发,且C型HBx比B型对细胞基本自噬的增加更加显著。【结论】HBV通过HBx增强细胞的基本自噬,且不同基因型HBx对细胞基本自噬的增强程度不同,为进一步阐明HBV感染机理奠定了基础。     

Antiviral Effect of Interferon-Induced Guanylate Binding Protein-1 against Coxsackie Virus and Hepatitis B Virus B3 in Vitro

Guanylate binding protein-1(GBP-1)is an interferon-induced protein.To observe its antiviral effect against Hepatitis B virus(HBV)and Coxsackie virus B3(CVB3),we constructed an eukaryotic expression vector of human GBP-1(hGBP-1).Full-length encoding sequence of hGBP-1 was amplified by long chain RT-PCR and inserted into a pCR2.1 vector,then subcloned into a pCDNA3.1(-)vector.Recombinant hGBP-1 plasmids and pHBV1.3 carrying 1.3-fold genome of HBV were contransfected into HepG2 cells,and inhibition effect of hGBP-1 against HBV replication was observed.Hela cells transfected with recombinant hGBP-1 plasmids were challenged with CVB3,and viral yield in cultures were detected.The results indicated that recombinant eukaryotic expression plasmid of hGBP-1 was constructed successfully and the hGBP-1 gene carried in this plasmid could be efficiently expressed in HepG2 cells and Hela cells.hGBP-1 inhibit CVB3 but not HBV replication in vitro.These results demonstrate that hGBP-1 mediates an antiviral effect against CVB3 but not HBV and perhaps plays an important role in the interferon-mediated antiviral response against CVB3.     

Antiviral Effect of Interferon-Induced Guanylate Binding Protein-1 against Coxsackie Virus and Hepatitis B Virus B3 in Vitro

Guanylate binding protein-1(GBP-1) is an interferon-induced protein. To observe its antiviral effect against Hepatitis B virus (HBV) and Coxsackie virus B3 (CVB3), we constructed an eukaryotic expression vector of human GBP-1(hGBP-1). Full-length encoding sequence of hGBP-1 was amplified by long chain RT-PCR and inserted into a pCR2.1 vector, then subcloned into a pCDNA3.1(-) vector. Recombinant hGBP-1 plasmids and pHBV1.3 carrying 1.3-fold genome of HBV were contransfected into HepG2 cells, and inhibition effect of hGBP-1 against HBV replication was observed. Hela cells transfected with recombinant hGBP-1 plasmids were challenged with CVB3, and viral yield in cultures were detected. The results indicated that recombinant eukaryotic expression plasmid of hGBP-1 was constructed successfully and the hGBP-1 gene carried in this plasmid could be efficiently expressed in HepG2 cells and Hela cells. hGBP-1 inhibit CVB3 but not HBV replication in vitro. These results demonstrate that hGBP-1 mediates an antiviral effect against CVB3 but not HBV and perhaps plays an important role in the interferon-mediated antiviral response against CVB3.