乙型肝炎病毒感染与肝细胞癌病例对照研究

原发性肝细胞癌(Hepatocellular Carcinoma,HCC)是我国乃至世界上最常见、最具有危害性的恶性肿瘤之一[1,2],我国每年新发病例约占全球45%,在恶性肿瘤的年死亡率分别占农村和城市恶性肿瘤死亡率的第一和第二位,严重危害劳动人民的健康与生命安全.本研究于2003年7-9月间对武汉地区原发性肝癌患者进行了危险因素调查,为探索病因和发病机制以及制定原发性肝癌的防治策略提供参考依据,以利于进一步做好市民的卫生保健工作.     

放射治疗肺癌患者的观察及护理

原发性支气管肺癌简称为肺癌.据近年来我国的调查,肺癌在一些城市和厂矿地区已占恶性肿瘤的首位,而且发病率和死亡率逐年上升.我科自2001年11月至2003年12月共收治肺癌患者23例,现对23例肺癌患者放射治疗的护理体会报告如下.     

营养与肿瘤

恶性肿瘤是严重危害人民生命与健康的常见病之一,一般死亡率为所有疾病死亡率的前三位,我国也是如此。在中国,恶性肿瘤在男性占笫二位,女性占第三位。肿瘤的发病率中以胃癌为最高,占所有肿瘤的1/4,其次是食管癌、肝癌、宫颈癌、肺癌、肠癌、白血病、鼻咽癌、乳腺癌等。以上9种占全部恶性肿癌发病率的90％,胃癌发病以男性为多,男性为女性的2.1倍。肿瘤的发病率及死亡率在30岁以前很低,35岁开始明显上升,随年龄的增加而增加。70岁以后变低,在中国肿瘤的平均发病年龄为62岁。在国内发病率最高的9种肿瘤中,几乎有一半,即胃癌,肝癌、食管癌、肠癌属于消化系     

恶性卵巢肿瘤的数学模型诊断

近年来恶性卵巢瘤的发病率具有逐年增高的趋势。据我院统计,恶性卵巢瘤的发病率占妇科恶性肿瘤的首位。由于恶性卵巢瘤早期往往无明显症状,而且与其他脏器的恶性肿瘤作比较,缺少早期诊断的方法。确诊时大约三分之二的病例已经处于晚期,结果五年存活率仅为25～30％,死亡率亦占妇科恶性肿瘤的首位。因此,如何提高恶性卵巢瘤的早期诊断成一重要课题。本文对我院妇产科近五年来收治的卵巢肿瘤采用回顾方法,     

卵巢恶性肿瘤的高危因素及治疗的研究进展

卵巢恶性肿瘤是女性生殖器常见的三大恶性肿瘤之一,也是人体病理类型最复杂的恶性肿瘤,发病率仅次于子宫颈癌和子宫体癌而列居第三位,死亡率居妇科恶性肿瘤首位.近年来,随着对卵巢恶性肿瘤高危因素、诊断、治疗及预后相关因素的进一步研究,该疾病的预后得到了显著改善,患者的生存率和生存质量显著提高.本文将对卵巢恶性肿瘤的高危因素及治疗的研究进展做一综述.     

肝癌专题文献

原发性肝癌是世界上最常见的恶性肿瘤之一,全球每年约有二十五万新发病人。近年来,发病率还在上升。在我国,肝癌占各种恶性肿瘤第3位,每年约有十万人死于该病。可见肝癌对人民健康的危害极其严重。开展肝癌防治研究,具有重大现实和战略意义。现把最新肝癌文献介绍如下:     

封面说明

正食管癌是常见的恶性肿瘤之一,死亡率位居我国恶性肿瘤死因的第4位。肿瘤转移是导致食管癌患者死亡的重要原因,深入研究食管癌侵袭和转移的分子机制,将有助于改善食管癌的治疗,降低患者的死亡率。由SERPINE1基因编码的纤溶酶原激活物抑制因子1(plasminogenactivator inhibitor-1,PAI-1)被报道在多种类型     

326 例眼睑肿物的临床病理分析

目的:总结眼睑肿物的临床病理类型及特点。方法:收集2000年1月至2011年6月到青岛大学医学院附属医院眼科住院部行手术治疗眼睑肿物患者326例的临床病理资料进行分析。结果:在326例眼睑肿物中,良性肿瘤156例,占47.9%,恶性肿瘤63例,占19.3%;炎性改变98例,占30.1%。良性肿瘤的前五位分别是色素痣、乳头状瘤、囊肿、疣、血管瘤;恶性肿瘤前三位分别是基底细胞癌、睑板腺癌、鳞状细胞癌;炎性改变以炎性肉芽肿最常见。儿童多发的眼睑肿物为皮样瘤、钙化上皮瘤。结论:眼睑病变以良性肿瘤多见,其次为炎性改变。良性肿瘤中以色素痣、乳头状瘤和囊肿多见;恶性肿瘤最常见的为基底细胞癌,儿童眼睑肿瘤以皮样瘤最多见。     

综合护理干预对鼻咽癌放疗患者的康复影响

鼻咽癌是我国常见的恶性肿瘤之一,在头颈部恶性肿瘤中占首位,好发于我国南方各省.鼻咽癌的治疗方法以放射治疗为首选.我科自2009年4月-2010年8月对收治的50例鼻咽癌患者进行直线加速器放疗,通过采取一系列的综合护理干预措施,取得良好效果,现报告如下.     

DKK-1 与肝脏恶性肿瘤的相关研究进展

肝脏恶性肿瘤包括原发性肝癌、继发性肝癌、肝母细胞瘤、肝脏淋巴瘤、肝脏血管内皮细胞肉瘤、纤维板层肝细胞癌、肝脏未分化胚胎肉瘤等发生在肝脏的恶性病变。其中原发性肝癌(primary liver cancer,PLC)是临床上最常见的恶性肿瘤之一。PLC在我国的发病人数占全球的55%,是我国第二个最常见的癌症死亡原因。由于肝脏恶性肿瘤具有隐匿性强、恶性程度高,病情进展快的特点,很多患者就诊时已到疾病中晚期,即使采取多学科综合治疗,预后也很不理想。因此,美国肝病研究学会(AASLD)和卫生部制定的《原发性肝癌诊疗规范(2011年版)》特别强调了早期筛查和早期监测对提高患者生存时间和生存质量的作用。甲胎蛋白(AFP)联合影像学检查是目前筛查肝脏恶性肿瘤的主要方法,但是AFP和影像学检查尚缺乏足够的敏感性和特异性,尤其对于早期癌症的诊断而言。DKK-1(dickkopf-1)是近年来由德国科学家新发现的一种分泌型糖蛋白。DKK-1与肝脏恶性肿瘤,尤其与原发性肝癌的早期诊断和预后判断关系密切,是最值得期待的肿瘤诊断标志物之一。本文谨对DKK-1的分子生物学特点、在恶性肿瘤中的表达以及与肝脏恶性肿瘤的关系进行综述,探讨其作为肝癌诊断蛋白标志物的研究现状及临床应用前景。     

Proteomic analysis of HBV-associated HCC: Insights on mechanisms of disease onset and biomarker discovery

The development of hepatocellular carcinoma (HCC) can be considered as an end-stage outcome of chronic hepatitis B virus (HBV) infection. Early prognostic markers are needed to allow effective treatments and prevent HCC from developing. Proteomics analysis has been used to identify markers from clinical samples from HCC patients. This approach can be further improved by identifying early biomarkers before the onset of HCC. One way would be to use the cell-based HBV replication system, which is reflective of the early stage of virus infection and thus secreted proteins identified at this stage may have relevance in HCC prognosis. In this review, we focus the discussion on the current status of proteomics analysis of cellular proteins and HCC biomarker identification, with a special highlight on the potential of the cell-based HBV replication system for the identification of prognostic HCC biomarkers.     

Distinct Distribution Pattern of Hepatitis B Virus Genotype C and D in Liver Tissue and Serum of Dual Genotype Infected Liver Cirrhosis and Hepatocellular Carcinoma Patients

Aims

The impact of co-infection of several hepatitis B virus (HBV) genotypes on the clinical outcome remains controversial. This study has for the first time investigated the distribution of HBV genotypes in the serum and in the intrahepatic tissue of liver cirrhotic (LC) and hepatocellular carcinoma (HCC) patients from India. In addition, the genotype-genotype interplay and plausible mechanism of development of HCC has also been explored.

Methods

The assessment of HBV genotypes was performed by nested PCR using either surface or HBx specific primers from both the circulating virus in the serum and replicative virus that includes covalently closed circular DNA (cccDNA) and relaxed circular DNA (rcDNA) of HBV from the intrahepatic tissue. The integrated virus within the host chromosome was genotyped by Alu-PCR method. Each PCR products were cloned and sequences of five randomly selected clones were subsequently analysed.

Results

HBV/genotype D was detected in the serum of all LC and HCC patients whereas the sequences of the replicative HBV DNA (cccDNA and rcDNA) from the intrahepatic tissue of the same patients revealed the presence of both HBV/genotype C and D. The sequences of the integrated viruses exhibited the solo presence of HBV/genotype C in the majority of LC and HCC tissues while both HBV/genotype C and D clones were found in few patients in which HBV/genotype C was predominated. Moreover, compared to HBV/genotype D, genotype C had higher propensity to generate double strand breaks, ER stress and reactive oxygen species and it had also showed higher cellular homologous-recombination efficiency that engendered more chromosomal rearrangements, which ultimately led to development of HCC.

Conclusions

Our study highlights the necessity of routine analysis of HBV genotype from the liver tissue of each chronic HBV infected patient in clinical practice to understand the disease prognosis and also to select therapeutic strategy.     

Polymorphism analyses of hepatitis B virus X gene in hepatocellular carcinoma patients from southern China

Chronic hepatitis B virus(HBV)infection is one of the major causes of hepatocellularcarcinoma(HCC),and the HBV X(HBx)gene plays a critical role in the molecular pathogenesis ofHBV-related HCC.We have investigated whether there are particular HBx gene mutations associated withHCC in patients from southern China.The HBx gene was examined in 51 paraffin-embedded tumor tissuesamples from patients with HCC and 25 serum samples from the HBV carrier by nested polymerase chainreaction(PCR),single-stranded conformational polymorphism and heteroduplex analysis.The HBx geneswith potentially important mutations from tumor tissue samples were cloned,sequenced and aligned withthe published HBx gene sequence.HBV genotypes in tumor tissue samples were analyzed by nested PCR.Analyses of HBx gene polymorphism showed that 31.3% of HBx gene fragments in tumor tissue sampleshad a special pattern.A common deletion at nt 382-400 of the HBx gene accompanied by 29 point mutationswas detected in four randomly selected tumor tissue samples with this pattern which caused a frame-shiftin the HBx open reading frame with a new stop codon at nt 1818,resulting in an HBx polypeptide chaintruncated at the C end in these cases.Among the four randomly selected samples,three were HBV genotypeB,and one was not detected by our present assay.In another tumor tissue sample,amplification of thefull-length HBx gene yielded a shorter fragment.Sequencing of this fragment revealed a 264 bp deletionbetween nt 1577 and 1840 of the HBV gene.These results suggest that HBx gene mutation occurs frequentlyin HCC samples,and the deletion at nt 382-400 of the HBx gene might play a role in carcinogenesis of HCCin southern China.     

Heat-shock protein 27: a potential biomarker for hepatocellular carcinoma identified by serum proteome analysis

Hepatocellular carcinoma (HCC) is the third leading cause of cancer mortality worldwide and ranks second in China. The prognosis of HCC remains dismal mainly because of its late diagnosis, especially in patients with coexisting chronic liver diseases. To identify serum biomarkers for HCC, sera from 20 healthy volunteers, 20 hepatitis B virus (HBV) infected patients and 20 HCC patients were selected for screening study and same number of sera into the same three groups were used for validation study. A strategy including sonication, albumin and immunoglobulin G (IgG) depletion and desalting was optimized for screening differentially expressed proteins of low abundance in serum. By 2-DE image analysis and MALDI-TOF-MS/MS identification, eight proteins including heat-shock protein 27 (HSP27), alpha-fetoprotein (AFP), alpha-1 antitrypsin, clusterin, caeruloplasmin, haptoglobin alpha2 chain, tranferrin and transthyretin were found significantly changed among the healthy, HBV and HCC groups. Further validation study by Western blot showed the detection of HSP27 in 90% HCC sera and two HBV sera, but in none of normal sera. Thus, 2-DE based serum proteome analysis can be useful in the screening of serum biomarkers for HCC and HSP27 could aid in the diagnosis of HCC though further validation is needed.     

Identification and extensive analysis of inverted-duplicated HBV integration in a human hepatocellular carcinoma cell line

Hepatitis B virus (HBV) DNA is often integrated into hepatocellular carcinoma (HCC). Although the relationship between HBV integration and HCC development has been widely studied, the role of HBV integration in HCC development is still not completely understood. In the present study, we constructed a pooled BAC library of 9 established cell lines derived from HCC patients with HBV infections. By amplifying viral genes and superpooling of BAC clones, we identified 2 clones harboring integrated HBV DNA. Screening of host-virus junctions by repeated sequencing revealed an HBV DNA integration site on chromosome 11q13 in the SNU-886 cell line. The structure and rearrangement of integrated HBV DNA were extensively analyzed. An inverted duplicated structure, with fusion of at least 2 HBV DNA molecules in opposite orientations, was identified in the region. The gene expression of cancer-related genes increased near the viral integration site in HCC cell line SNU-886.     

IL-23 production of liver inflammatory macrophages to damaged hepatocytes promotes hepatocellular carcinoma development after chronic hepatitis B virus infection

Liver inflammation after chronic hepatitis B virus (HBV) infection is essential for hepatocellular carcinoma (HCC) development. We did a nested case-control study based on QBC chronic HBV infection cohort to identify HCC-related inflammatory cytokines. Serum levels of distinct Th-cell representative cytokines at varied periods before HCC diagnosis were determined in 50 HCC cases and 150 age- and gender-matched controls who did not develop HCC in 8–10?years. The individuals with HCC outcome had statistically higher serum levels of IL-23 than controls (P?<?0.01). Further analysis in HCC tissues showed that CD14⁺ inflammatory macrophages were the major IL-23 producers. Monocytes-derived macrophages generated more amount of IL-23 after being stimulated with cell-associated HBV core antigen from damaged HBV-infected hepatocytes than the cells being stimulated with HBV-S and HBV e antigen, which are secreted from infected hepatocytes. IL-23 upregulated IL-23 receptor expressions on macrophages, enhanced macrophage-mediated angiogenesis. In HBV-transgenic (Alb1HBV) mice, administration of diethylnitrosamine induced more liver tumors than in wild-type mice. The livers of Alb1HBV mice had higher concentrations of IL-23 and vascular endothelial growth factor (VEGF) than the wild-type mice. Neutralizing IL-23 activity, diethylnitrosamine-treated Alb1HBV mice developed significantly less tumors and produced less VEGF, tumor angiogenesis was inhibited with dramatically decreased CD31⁺ cells within tumor mass (all P?<?0.01).

Genotype-specific genomic markers associated with primary hepatomas, based on complete genomic sequencing of hepatitis B virus

We aimed to identify genomic markers in hepatitis B virus (HBV) that are associated with hepatocellular carcinoma (HCC) development by comparing the complete genomic sequences of HBVs among patients with HCC and those without. One hundred patients with HBV-related HCC and 100 age-matched HBV-infected non-HCC patients (controls) were studied. HBV DNA from serum was directly sequenced to study the whole viral genome. Data mining and rule learning were employed to develop diagnostic algorithms. An independent cohort of 132 cases (43 HCC and 89 non-HCC) was used to validate the accuracy of these algorithms. Among the 100 cases of HCC, 37 had genotype B (all subgenotype Ba) and 63 had genotype C (16 subgenotype Ce and 47 subgenotype Cs) HBV infection. In the control group, 51 had genotype B and 49 had genotype C (10 subgenotype Ce and 39 subgenotype Cs) HBV infection. Genomic algorithms associated with HCC were derived based on genotype/subgenotype-specific mutations. In genotype B HBV, mutations C1165T, A1762T and G1764A, T2712C/A/G, and A/T2525C were associated with HCC. HCC-related mutations T31C, T53C, and A1499G were associated with HBV subgenotype Ce, and mutations G1613A, G1899A, T2170C/G, and T2441C were associated with HBV subgenotype Cs. Amino acid changes caused by these mutations were found in the X, envelope, and precore/core regions in association with HBV genotype B, Ce, and Cs, respectively. In conclusion, infections with different genotypes of HBV (B, Ce, and Cs) carry different genomic markers for HCC at different parts of the HBV genome. Different HBV genotypes may have different virologic mechanisms of hepatocarcinogenesis.     

iTRAQ‐coupled 2‐D LC‐MS/MS analysis of protein profile associated with HBV‐modulated DNA methylation

The development of hepatocellular carcinoma (HCC) is believed to be associated with multiple risk factors, including the infection of hepatitis B virus (HBV). Based on the analysis of individual genes, evidence has indicated the association between HCC and HBV and has also been expanded to epigenetic regulation, with an involvement of HBV in the DNA methylation of the promoter of cellular target genes leading to changes in their expression. Proteomic study has been widely used to map a comprehensive protein profile, which in turn could provide a better understanding of underlying mechanisms of disease onset. In the present study, we performed a proteomic profiling by using iTRAQ‐coupled 2‐D LC/MS‐MS analysis to identify cellular genes down‐regulated in HBV‐producing HepG2.2.15 cells compared with HepG2 cells. A total of 15 proteins including S100A6 and Annexin A2 were identified by our approach. The significance of these cellular proteins as target of HBV‐mediated epigenetic regulation was supported by our validation assays, including their reactivation in cells treated with 5‐aza‐2′‐deoxycytidine (a DNA methyltransferase inhibitor) by real‐time RT‐PCR and Western blot analysis, as well as the DNA methylation status analysis by bisulfite genome sequencing. Our approach provides a comprehensive analysis of cellular target proteins to HBV‐mediated epigenetic regulation and further analysis should facilitate a better understanding of its involvement in HCC development.     

Enhanced expression of matrix metalloproteinase-9 by hepatitis B virus infection in liver cells

The hepatitis B virus (HBV) is a major cause of human liver disease, including hepatocellular carcinoma (HCC). The prognosis for HCC is largely dependent on the clinicopathological characteristics regarding invasion and metastasis. Enhanced matrix metalloproteinase-9 (MMP-9) expression has been implicated as playing an important role in metastasis and invasion of HCC. However, the relationship between HBV infection and MMP-9 expression in HCC is currently poorly understood. We report here on a study of the levels of MMP-9 and MMP-2 expression in human fetal liver tissue, rat liver tissue, and Chang, HepG2, and Hep3B cells by gelatin zymography. Among these sources, Hep3B cells, which contain the integrated hepatitis B viral genome, continuously secrete the hepatitis B viral surface antigen, and express HBV genomic RNA, expressed high levels of proMMP-9, and a small amount of active MMP-9 was detected in Hep3B cells as assayed by zymography. We investigated the issue of whether HBV infection affects MMP-9 expression, which is known to play an important role in HCC invasion and metastasis. As a first step, human fetal hepatocyte (HFH) and HepG2 (HCC origin, HBV not detected) cells were subjected to infection with HBV, and the resulting infected cells successfully established are hereafter referred to as HFH-T2 and HepG2-HBV. The expression of MMP-9 was upregulated by the infected HBV in HFH-T2 and HepG2-HBV cells, as assayed by zymography, Northern blot, and Western blot analysis, and small amounts of active MMP-9 were detected in HFH-T2 and HepG2-HBV cells as assayed by zymography. The activation of the immature proMMP-9 to the mature MMP-9 could be induced by plasmin treatment. The activation of proMMP-9 was increased to a greater extent with plasmin treatment than without plasmin in HFH-T2 and HepG2-HBV cells but the addition of recombinant TIMP-1 inhibited the activation of proMMP-9. Finally, the addition of plasmin to the invasion assay using Matrigel resulted in an increase in invasiveness of HFH-T2 and HepG2-HBV cells, as well as MMP-9 activation, but the treatment with TIMP-1 inhibited the invasiveness of HFH-T2 and HepG2-HBV cells as well as MMP-9 activation. We conclude from these findings that HBV infection of hepatocytes and HepG2 cells affected the upregulation of MMP-9 expression and MMP-9 activation and, thus, increased the invasion potential by plasmin. To our knowledge, this is a first report showing that an HBV infection is linked to the upregulation of MMP-9 in HCC.