我国九个地区丁型肝炎病毒感染状况及其与乙型肝炎病毒复制指标关系的研究

对１５４５例各类乙型肝炎病毒表面抗原（ＨＢｓＡｇ）阳性肝病和无症状ＨＢｓＡｇ携带者的血清进行了乙型肝炎病毒（ＨＢＶ）与丁型肝炎病毒（ＨＤＶ）感染标记物的测定。结果表明，ＨＤＶ感染率为１３．０１％，其中ＨＤＡｇ和抗－ＨＤ阳性率分别为２．９１％和１０．０９％。而且在全国九个地区均有ＨＤＶ感染者存在，说明其分布是较为广泛的。同时还表现出，男性高于女性，慢性肝炎、重型肝炎及原发性肝癌高于急性肝炎和无症状ＨＢｓＡｇ携带者。提示ＨＢＶ与ＨＤＶ合并感染或重叠感染可能导致病情加重和感染的慢性化。本项研究结果还揭示，在ＨＢＶ与ＨＤＶ合并或重叠感染时，可能对ＨＢＶ的复制指标（ＨＢｅＡｇ·ＨＢＶＤＮＡ）有一定的抑制现象。     

Polymorphisms in the promoter regions of the matrix metalloproteinases-7, -9 and the risk of endometriosis and adenomyosis in China

Matrix metalloproteinases (MMPs) may contribute to the development of endometriosis. The aim of this study was to assess the effects of the polymorphisms in the promoters of MMP-7 (181A/G) and MMP-9 (1562C/T) on the risk of occurrence of endometriosis and adenomyosis. We genotyped 219 patients (143 women with endometriosis, 76 women with adenomyosis) and 160 control women in North China. There was a significant difference in frequency of the MMP-7 genotype between endometriosis and controls (P = 0.01) and also between adenomyosis and controls (P = 0.01). The frequency of the G allele in two groups of patients (7.3 and 7.9%) was significantly higher than in the controls (2.8%) (P = 0.01 and 0.01, respectively). Compared to the A/A genotype, the genotype with the -181G allele showed a significantly increased susceptibility to both diseases, with adjusted odds ratio of 2.62 [95% confidence interval (CI) = 1.17-5.87] for endometriosis and 3.14 (95% CI = 1.26-7.81) for adenomyosis. However, the overall genotype and allelotype distribution of the MMP-9 in the two case groups were not different from that of controls. We conclude that MMP-7-181A/G polymorphism has a potential to be a susceptibility factor for endometriosis and adenomyosis while MMP-9-1562C/T polymorphism may not provide a useful marker to predict susceptibility to endometriosis and adenomyosis, at least in women from North China.     

Changes in Hepatitis C Virus Genotype Distribution in Chronic Hepatitis C Infection Patients

Purpose: Identification of hepatitis C virus (HCV) genotypes is very important in the selection of antiviral treatment, dose adjustment of antiviral agents, determining the treatment duration and following-up of treatment response. We aimed to determine the distribution pattern of HCV genotypes in chronic hepatitis C infection (CHC) patients. Materials and Methods: We have included 106 CHC patients who were positive in the anti-HCV and HCV-RNA tests performed in our hospital during the 16-month period. Anti-HCV assays were performed on device using a chemiluminescent microparticle immunoassay, while HCV-RNA tests and HCV genotyping assays were performed by real-time polymerase chain reaction. Results: Of the 106 cases; genotype 1b was detected in 67.0%, genotype 3 was detected in 16.0%, genotype 1a was detected in 14.2% and genotype 2 was detected in 2.8% patients. Genotypes 4, 5 and 6 were not detected in our study group. There were no statistically significant differences between the gender and age groups according to the HCV genotype distribution. The genotype 3 detection rate (16%) was the highest rate among the studies compared with the other studies in our country. Conclusions: Events that cause social changes such as war and immigration and intense commercial and touristic activities affect and alter the HCV genotype distribution in HCV-infected patients. For this reason, further multicentre studies are required reflecting all the regions in order to determine the genotype distribution in HCV-infected patients at regular intervals.     

用聚合酶链反应技术研究重型肝炎患者乙型和丙型肝炎病毒的混合感染

采用聚合酶链反应技术（ＰＣＲ）检测了３６例重型肝炎患者血清中乙型肝炎病毒（ＨＢＶ）和丙型肝炎病毒（ＨＣＶ）的复制状况，发现较普遍存在ＨＢＶ复制，阳性率高达７８％；ＨＣＶ复制亦较活跃，为６１％。提示病毒因素仍然是引起肝衰竭的重要原因。临床资料表明，同时感染两种病毒的重肝患者（４１％）预后较差，ＨＢＶ引起的肝坏死较ＨＣＶ引起的更为严重，两者在体内复制有互相抑制作用，但合并感染可造成肝脏的持续损伤。     

乙型和丙型肝炎病毒感染的免疫学研究进展

世界范围内持续感染乙型肝炎病者（HBV）、丙型肝炎病毒（HCV）的患者已超过5亿。这两种病毒在病原学上有许多共同特点，但在病毒学特点上以及在慢性肝炎的免疫逃逸机制上均有着显著的差别。在早期天然免疫应答方面，HBV感染最初几周并不引起肝脏基因表达的改变，而HCV则会引起许多肝内基因表达改变。在特异性免疫应答方面，HBV和HCV感染后机体清除病毒的途径主要有特异性CTL细胞的杀伤作用、CD4^＋细胞的辅助作用、抗体的中和作用或杀伤作用。相对于HBV来说，HCV在成人更多引起慢性肝炎。     

补体C6 T1674C单核苷酸多态性在中国汉族人群中的分布

目的获得人类补体第六成分(C6)T1674C多态性在中国汉族人群的分布特征。方法采用聚合酶链反应-单链构象多态性(polymerase chain reaction-single strand conformation polymorphism PCR-SSCP)结合核酸测序方法检测150例中国四川泸州汉族人群C6基因T1674C的基因型频率及等位基因频率。结果基因型TT、TC、CC的频率分别为44.67%、23.33%、32.00%,等位基因T、C频率分别为56.33%和43.67%,其基因型频率分布符合Hardy-weinberg平衡定律,个人识别机率为61.7%。结论中国汉族人群C6基因T1674C多态性个人识别能力高,重复性好,在人类遗传学研究及法医学个人识别中有较好的应用价值。     

应用非同位素原位杂交检测肝组织中丙型肝炎病毒基因的分布

为了研究丙型肝炎患者肝组织中丙型肝炎病毒（ＨＣＶ）基因的分布，我们应用地高辛标记的ＨＣＶ基因５＇端非翻译区的探针（长度为３２个寡核苷酸），对２４例急、慢性丙型肝炎患者活检的肝组织石蜡包埋切片进行了原位核酸杂交检测。结果显示：ＨＣＶ基因阳性的肝组织标本有１１例，检出率是４５．８％（１１／２４）。ＨＣＶ基因主要分布于肝细胞浆，偶见于肝细胞核内。此外在肝血窦的ｋｕｐｆｆｅｒ细胞、小血管内皮细胞和汇管区附近均有明显的ＨＣＶ基因阳性染色，而对照组均未发现ＨＣＶ基因阳性信号。     

乙型肝炎患者及其子女HBV DNA U5样序列单核苷酸多态性分析

目的进一步探讨乙型肝炎在患者及其亲子代之间的垂直传播。方法应用单核苷酸多态性(SNP)、聚合酶链反应-单链构象多态性(PCR-SSCP)分析等分子遗传学技术和方法检测了乙型肝炎患者家系30个,68例受试者。结果U5样序列PCR检测结果表明,游离型和整合型HBV DNA在乙型肝炎患者(HBP)与其发病后出生子女(HBPa)实验组检出率之间一致性增高,其检出率分别与乙型肝炎患者发病前出生子女(HBPb)和正常对照组的比较,差异均显著,P<0.05。乙型肝炎患者父子之间的SNP分析发现,在U5样序列区和非U5样序列区多个碱基位点处出现碱基替换、插入或缺失,1 908A→T1、950 G→T1、967 T→C,还存在1 900T缺失和1 903C插入等。乙型肝炎患者父子之间的SNP在1 9081、9501、9671、900和1 903位点一致。结论乙型肝炎可以在HBsAg阳性的男性乙型肝炎患者(MHBP)及其子女之间遗传传递,为乙型肝炎的遗传传递进一步提供了分子遗传学证据。     

Contribution of Toll-Like Receptor 9 Gene Single-Nucleotide Polymorphism to Systemic Lupus Erythematosus in Egyptian Patients

Background: Systemic lupus erythematosus (SLE) is an autoimmune disease, with multiple genetic and environmental factors involved in its etiology. The toll-like receptor 9 (TLR9) gene has been reported to have important roles in the development and progression of SLE. In this case-control study, the effect of TLR9 polymorphism on susceptibility to SLE was investigated in Egyptian patients.

Methods: We studied the distribution of the TLR9 rs352139 (G + 1174A) single nucleotide polymorphism (SNP) by allele-specific polymerase chain reaction (PCR) in 104 Egyptian patients with SLE and 108 age-, sex-, and ethnically matched controls.

Results: There was no statistically significant difference in the distribution of the AA genotype and alleles between SLE patients and the control group in our study; however, the GA heterozygous patients were three times more likely to develop SLE (P < 0.001). A significant association was detected between TLR9 genotypes and some of the disease manifestations as myositis (p = 0.032), psychosis (p = 0.014), photosensitivity (p = 0.002), and pleurisy (p = <0.001). Moreover, we observed a significant association between the TLR9 AA and GA genotypes and the presence of antinuclear antibodies (ANA) (p = 0.038).

Conclusion: The G + 1174A SNP in the toll receptor 9 gene may contribute to the genetic susceptibility of SLE in Egyptian patients. Also, an influence for this polymorphism on disease manifestations has been elucidated.     

几种基因的单核苷酸多态性与类风湿关节炎

单核苷酸多态性(SNPs)是新一代的基因分子标记,随着人类基因组图谱的绘制成功,SNPs被应用于寻找各种致病基因。类风湿关节炎(RA)是一种全身自身免疫性疾病,到目前为止,其发病机制尚未完全清楚。本文简要介绍了SNPs及几种基因的SNPs与RA的关系。     

Evaluation of Clinical Value of Single Nucleotide Polymorphisms of Dihydropyrimidine Dehydrogenase Gene to Predict 5-Fluorouracil Toxicity in 60 Colorectal Cancer Patients in China

Dihydropyrimidine dehydrogenase (DPD) activity could be affected by single nucleotide polymorphisms (SNPs), resulting in either no effect, partial or complete loss of DPD activity. To evaluate if SNPs of DPD can be used to predict 5-FU toxicity, we evaluated five SNPs of DPD (14G1A, G1156T, G2194A, T85C and T464A) by TaqMan real time PCR in 60 colorectal cancer patients. Clinical data demonstrated that there was higher correlation between DPD activity and toxic effects of 5-FU (p<0.05). Six patients were positive for G2194A detection, which were all heterozygous. Two patients had lower DPD activities (< 3) with higher toxic effects (≥stage III) while one patient was also positive for T85C detection. Ten patients were positive for T85C detection. Two patients were homozygous with lower DPD activities and higher toxic effects. Two patients were positive for the T464A detection, which were heterozygous with lower DPD activity and higher toxic effects and also positive for T85C detection. These data clearly indicated that the T464A and homozygous of the T85C are stronger biomarkers to predict the 5-FU toxicity. Our study significantly indicated that the detection for G2194A, T85C and T464A could predict ~13% of 5-FU severe toxic side effects.     

HRMA技术在检测线粒体DNA单核苷酸多态性中的应用

目的 高分辨熔解曲线分析（high-resolution melting analysis,HRMA）方法是通过检测和分析不同基因型DNA分子在解链时随荧光信号变化而产生的不同熔解曲线,进行SNPs分型的技术。探索HRMA技术在检测线粒体DNA分子基因多态性中的应用。方法 利用未标记探针（unlabled probe）HRMA技术,进行人线粒体DNA 单核苷酸多态性(mtSNPs)分型,并经限制性片段长度多态(restriction fragment length polymorphism, RFLP)分型技术和测序验证。结果 结果显示,未标记探针HRMA分型与RFLP分型和测序验证结果完全相符。结论 本研究表明未标记探针HRMA技术方法准确率较高,是一种适合mtSNPs分型的方法。     

Distribution of Hepatitis C Virus Genotypes in Istanbul,Turkey

Purpose: The hepatitis C virus (HCV) has seven main genotypes and multiple subtypes. The distribution of HCV genotypes varies across geographical regions worldwide. Updated estimates of HCV genotype distributions have a critical importance for developing strategies to manage or eliminate HCV infection. The aim of this study was to determine the distribution of HCV genotypes in patients with HCV admitted to a university hospital in Istanbul, Turkey. Materials and Methods: A total of 412 HCV RNA positive patients with 46.6% of males and 53.4% of females between January 2013 and September 2016 were included in the study. Genotyping of HCV of the study population was performed by a commercial reverse hybridisation line probe-based assay. Results: Genotype 1 (82.5%) was dominant genotype, followed by genotype 3 (10.7%), genotype 2 (4.6%) and genotype 4 (2.2%). Among patients with genotype 1, subtype 1a, 1b and undetermined subtype were 6.3%, 38.8% and 37.4%, respectively. It was observed that genotype proportion was dependent on gender and age of the patients. Genotype 1 and genotype 2 were more prevalent in females, whereas genotypes 3 and 4 were more prevalent in males. Genotype 1 in the older patients and genotype 3 in the younger patients were more prevalent. Conclusion: The majority of patients with HCV infection had genotype 1 (82.5%), followed by genotype 3, 2 and 4. Monitoring the change in HCV genotype distribution is critical for the development of effective strategies for HCV elimination.     

丙型肝炎病毒NS3蛋白单克隆杂交瘤细胞株的建立及初步鉴定

应用国产基因工程表达的丙型肝炎病毒（ＨＣＶ）ＮＳ３区抗原免疫小鼠，然后取其脾细胞与小鼠骨髓瘤细胞系ＳＰ２／０融合，筛选出４株稳定分泌抗ＨＣＶＮＳ３区蛋白单克隆抗体（ＭｃＡｂ）的杂交瘤细胞株，分别命名为２Ｂ６，２Ｆ３，３Ｄ８，３Ｄ９，经初步研究表明这４株单抗与ＮＳ３抗原具有良好的反应性，与ＨＣＶ核心区多肽及乙型肝炎病毒表面抗原和ｅ抗原均无反应。抑制实验表明这４株抗体分别针对ＮＳ３抗原分子上的２个不同的抗原决定簇。     

丙型肝炎病毒核心区抗体分型方法的研究

用丙型肝炎病毒核心区２个型特异性俣成肽作为包被抗原，建立了ＥＬＩＳＡ法，对血清中抗－ＨＣＶ进行分型。在１２１份抗－ＨＣＶ阳性血清中检出ＣＰ９阳性率为８３．４７％，在１０１份ＣＰ９阳性血清中进行核心区抗体分型，其中血清１型１７．８２％，血清２型２５．７４％、１．２混合型７．９２％。     

利用双扩增聚合酶链反应检测非甲非乙型肝炎患者血清中丙型肝炎病毒RNA

利用丙型肝炎病毒（ＨＣＶ）５’－端序列合成两对引物，建立了灵敏、特异的ＨＣＶＲＮＡ双扩增聚合酶链反应检测方法。用此方法及第二代Ａｂｂｏｔｔ酶联抗－ＨＣＶ检测试剂盒，检测了４４例非甲非乙型肝炎患者血清及１０名抗－ＨＣＶ阴性健康人。在４４例患者中，４１例（９３％）ＨＣＶＲＮＡ阳性，３６例（８２％）抗－ＨＣＶ阳性，３３例（７５％）ＨＣＶＲＮＡ、抗－ＨＣＶ全部阳性。３例ＨＣＶＲＮＡ阴性，但抗－ＨＣＶ阳性，另外，有８例抗－ＨＣＶ阴性，ＨＣＶＲＮＡ阳性。１０名健康人ＨＣＶＲＮＡ均为阴性。结果表明，大部分（９２％）抗－ＨＣＶ阳性患者带有ＨＣＶ，但为了检测所有病毒血症患者，抗－ＨＣＶ检测是不够的，利用双扩增ＰＣＲ方法检测ＨＣＶＲＮＡ对于抗－ＨＣＶ阴性患者的诊断是非常有用的。     

Autophagy-Related 5 Gene rs510432 Polymorphism Is Associated with Hepatocellular Carcinoma in Patients with Chronic Hepatitis B Virus Infection

Background: Despite the identification of autophagy-related protein 5 (ATG5) as a molecule involved in the activated autophagy machinery during hepatitis B virus (HBV) infection and hepatocarcinogenesis, the consequences of ATG5 mutation carriage for patients with chronic HBV infection remain unclear. This study examined the association of ATG5 polymorphisms with HBV-related diseases including hepatocellular carcinoma (HCC).

Patients and Methods: Two functionally relevant polymorphisms ATG5 rs573775 and rs510432 were genotyped by ligase detection reaction-polymerase chain reaction in 403 patients with chronic HBV infection (171 chronic hepatitis, 119 cirrhosis and 113 HCC) and 196 healthy controls. Univariate and multivariate logistic regression was performed to evaluate factors associated with HCC.

Results: The rs573775 genotype and allele frequencies had no significant differences between patients with different clinical diseases. However, HCC patients had significantly higher frequency of rs510432 genotype AA (odds ratio [OR] 2.185, 95% confidence interval [CI] 1.042–4.581, P = 0.037, P value by Bonferroni correction [P_c] = 0.074) and allele A (OR 1.435, 95% CI 1.023–2.013, P_c = 0.036) than chronic hepatitis patients. In multivariate analyses, rs510432 allele A-containing genotypes (AA+GA) were independently associated with cirrhosis in comparison to chronic hepatitis (OR 1.927, 95%CI 1.011–3.017, P = 0.032). The rs510432 genotypes AA+GA were also independently associated with HCC in comparison to chronic hepatitis (OR 2.583, 95% CI 1.025–3.911, P = 0.006) or chronic HBV infection without HCC (OR 2.632, 95% CI 1.067–3.482, P = 0.032).

Conclusion: These results indicate that rs510432 genotypes AA+GA are associated with disease progression and HCC risk in chronic HBV infection, providing novel evidence for a role of ATG5 in the pathogenesis of HBV-related HCC.

Abbreviations: HBV: hepatitis B virus; HCC hepatocellular carcinoma; TNFSF10: tumor necrosis factor superfamily member 10; ATG5: autophagy-related protein 5; DNA: deoxyribonucleic acid; LDR-PCR: ligase detection reactions-polymerase chain reaction; PCR: polymerase chain reaction; SLE: systemic lupus erythematosus; BD: Behçet’s disease; IL-10: interlukin-10; LPS: lipopolysaccharide; PBMC: peripheral blood mononuclear cells; CWP: coal workers’ pneumoconiosis; TNF-α: tumor necrosis factor-α     

澳门地区丙型肝炎病毒基因型分析

应用逆转录－ＤＮＡ聚合酶链反应及ＤＮＡ序列分析，对采自澳门地区４３例抗－丙型肝炎病毒抗体阳性的慢性肝火患者及５５例血液透析患者血清进行了ＨＣＶ ＲＮＡ检测及基因型分析。