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Immunohistochemistry panel segregates molecular types of hepatocellular carcinoma in Brazilian autopsy cases

World Journal of Gastroenterology 2016年第27期22卷 6246-6256页

作者： Aloísio Felipe-Silva Alda Wakamatsu Cinthya dos Santos Cirqueira Venancio Avancini Ferreira Alves LIM 14-Patologia Hepatica Faculdade de Medicina da USP01246-903 Sao Paulo SPBrazil department of Pathology Medical Research Laboratory LIM14University of Sao Paulo School Of Medicine01246-903 Sao Paulo SPBrazil

AIM: To assess the distribution of proteins coded by genes reported as relevant for the molecular classification of hepatocellular carcinoma (HCC).METHODS: In this retrospective cross-sectional study, the following clinicopathological data were analyzed in 80 autopsied HCC patients: sex, age, ethnicity, alcohol intake, infection with hepatitis B and/or C virus, infection with human immunodeficiency virus, prior treatment, basic and immediate causes of death, liver weight, presence of cirrhosis, number and size of nodules, gross pattern, histological grade and variants, architectural pattern, invasion of large veins, and presence and location of extrahepatic metastases. The protein products of genes known to be involved in molecular pathogenesis of HCC, including epidermal growth factor receptor (EGFR), MET, keratin 19 (K19), vimentin, beta-catenin, mechanistic target of rapamycin (mTOR), extracellular signaling-related kinase (ERK)1, ERK2, Ki67, cyclin D1, caspase 3 and p53, were detected by immunohistochemistry on tissue microarrays. The expression levels were scored and statistically assessed for correlation with HCC ***: Infection with hepatitis C virus was identified in 49% of the 80 autopsy patients, cirrhosis in 90%, advanced tumors in 95%, and extrahepatic metastases in 38%. Expression of K19, p53 and ERK1 correlated to high-grade lesions. Expression of ERK1, nuclear beta-catenin, cyclin D1 and ERK2 correlated to higher rates of cell proliferation as determined by Ki67. Expression of MET, EGFR (> 0) and caspase 3 correlated with lower histological grades. Expression of EGFR correlated to that of caspase 3, and overexpression of EGFR (≥ 200/300) was observed in low-grade tumors more frequently (grades 1 and 2: 67% vs grade 3: 27% and grade 4: 30%). Expression of ERK1 was associated with that of K19 and vimentin, whereas expression of ERK2 was associated with that of cyclin D1, MET and membrane beta-catenin. Expression of vimentin

关键词： Hepatocellular carcinoma Epidermal growth factor receptor Autopsy Immunohistochemistry Liver Classification

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Dual gRNAs guided CRISPR/Cas9 system inhibits hepatitis B virus replication

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World Journal of Gastroenterology 2015年第32期21卷 9554-9565页

作者： Jie Wang Zhong-Wei Xu Shuang Liu Rui-Yang Zhang Shan-Long Ding Xiao-Meng Xie Lu Long Xiang-Mei Chen Hui Zhuang Feng-Min Lu State Key Laboratory of Natural and Biomimetic Drugs Department of Microbiology and Infectious Disease Center School of Basic Medicine Peking University Health Science Center department of Gastroenterology Pennsylvania Hospital University of Pennsylvania Beijing Artificial Liver Treatment and Training Center Beijing Youan Hospital Capital Medical University department of Epidemiology and Biostatistics College of Public Health Zhengzhou University

AIM: To screen and investigate the effective g RNAs against hepatitis B virus(HBV) of genotypes ***: A total of 15 g RNAs against HBV of genotypes A-D were designed. Eleven combinations of two above g RNAs(dual-g RNAs) covering the regulatory region of HBV were chosen. The efficiency of each g RNA and 11 dual-g RNAs on the suppression of HBV(genotypes A-D) replication was examined by the measurement of HBV surface antigen(HBs Ag) or e antigen(HBe Ag) in the culture supernatant. The destruction of HBV-expressing vector was examined in Hu H7 cells co-transfected with dual-g RNAs and HBVexpressing vector using polymerase chain reaction(PCR) and sequencing method, and the destruction of ccc DNAwas examined in Hep AD38 cells using KCl precipitation, plasmid-safe ATP-dependent DNase(PSAD) digestion, rolling circle amplification and quantitative PCR combined method. The cytotoxicity of these g RNAs was assessed by a mitochondrial tetrazolium ***: All of g RNAs could significantly reduce HBs Ag or HBe Ag production in the culture supernatant, which was dependent on the region in which g RNA against. All of dual g RNAs could efficiently suppress HBs Ag and/or HBe Ag production for HBV of genotypes A-D, and the efficacy of dual g RNAs in suppressing HBs Ag and/or HBe Ag production was significantly increased when compared to the single g RNA used alone. Furthermore, by PCR direct sequencing we confirmed that these dual g RNAs could specifically destroy HBV expressing template by removing the fragment between the cleavage sites of the two used g RNAs. Most importantly, g RNA-5 and g RNA-12 combination not only could efficiently suppressing HBs Ag and/or HBe Ag production, but also destroy the ccc DNA reservoirs in Hep AD38 ***: These results suggested that CRISPR/Cas9 system could efficiently destroy HBV expressing templates(genotypes A-D) without apparent cytotoxicity. It may be a potential approach for eradication of persistent HBV ccc DNA in c

关键词： Dual g RNAs CRISPR/Cas9 hepatitis B ccc DNA Antivi

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Usefulness of noninvasive transient elastography for assessment of liver fibrosis stage in chronic hepatitis C

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World Journal of Gastroenterology 2006年第48期12卷 7768-7773页

作者： Tadashi Takeda Takahiro Yasuda Yuji Nakayama Mika Nakaya Megumi Kimura Mariko Yamashita Ayumi Sawada Koji Abo Setsuko Takeda Hiroki Sakaguchi Susumu Shiomi Hitoshi Asai Shuichi Seki department of Hepatology Graduate School of Medicine Osaka City University Osaka Japan Central Clinical Laboratory Osaka City University Medical School Hospital Osaka Japan department of Nuclear Medicine Graduate School of Medicine Osaka City University Osaka Japan Hitoshi Asai Health Service Center Osaka Kyoiku University Osaka Japan

AIM：To evaluate the method of noninvasive transient elastography for assessment of histological stage of liver fibrosis in patients with chronic hepatitis C （CHC）.METHODS： Two hundred and thirty-seven patients with CHC were included in this study. Liver biopsy was performed under ultrasonography on 217 of the patients, excluding twenty with clear clinical evidence of liver cirrhosis. Fifty subjects without liver disease were enrolled as a control group （stage 0）. Twentyfive patients with sustained virological response （SVR） to interferon （IFN） therapy were also enrolled. These patients underwent liver biopsy before IFN therapy. Examination of liver stiffness （LS） was performed by elastography.RESISTS： Medians （50% levels） of LS were 4.1 （3.5-4.9）, 6.3 （4.8-8.5）, 8.8 （6.8-12.0）, 14.6 （10.5-18.6）, and 22.2 （15.4-28.0）, respectively, in the fibrosis stages 0-4 （P 〈 0.001）. LS was significantly correlated with four serum fibrosis markers. LS values in patients with SVR were 3.8 （3.5-5.6）, 5.2 （4.4-6.8）, 6.8 （6.1-7.6）, and 6.1 （3.6-7.9）, respectively, in the fibrosis stages 1-4. In all stages, LS for patients with SVR was significantly lower than that for patients who did not undergo IFN therapy. LS was significantly correlated with serum concentrations of hyaluronic acid, type Ⅳ collagen, type Ⅳ collagen 7S, and type Ⅲ procollagen N peptide.CONCLUSION： LS correlated well with the histological stage of fibrosis. Changes in liver fibrosis stage may thus be estimated noninvasively using transient elastography.

关键词： hepatitis C virus Echography Fibrosis Stiffness Interferon

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hepatitis C virus,mitochondria and auto/mitophagy:Exploiting a host defense mechanism

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World Journal of Gastroenterology 2014年第10期20卷 2624-2633页

作者： Vitalba Ruggieri Carmela Mazzoccoli Valerio Pazienza Angelo Andriulli Nazzareno Capitanio Claudia Piccoli Laboratory of Preclinical and Translational Research IRCCS Referral Cancer Center of Basilicata 85028 Rionero in Vulture (PZ) Italy Gastroenterology Unit IRCCS "Casa Sollievo della Sofferenza" Hospital 71013 Foggia Italy department of Medical and Experimental Medicine University of Foggia 71122 Foggia Italy

hepatitis C virus(HCV)is the major reason for liver transplantation and the main cause of liver-related morbidity and mortality in a great number of *** for the other viruses,this pathogen interferes in more than one process and in more than one way with host cell biology.A mounting body of evidence points,in particular,toward the drastic alterations of mitochondrial physiology and functions that virus is able to induce,albeit the mechanisms have partly remained *** of the mitochondria in immunity and in quality control systems,as autophagy,as well as the strategies that HCV has evolved to evade and even to manipulate mitochondrial surveillance for its benefit,highlights the importance of deepening the mechanisms that modulate this virus-mitochondrion interaction,not only to intensify our knowledge of the HCV infection pathogenesis but also to design efficient antiviral strategies.

关键词： hepatitis C virus Mitochondrial dysfunctions Immunity Autophagy Mitophagy

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Last line therapy with sorafenib in colorectal cancer: A retrospective analysis

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World Journal of Gastroenterology 2016年第23期22卷 5400-5405页

作者： Ksenia Martchenko Irene Schmidtmann Thomas Thomaidis Verena Thole Peter R Galle Marc Becker Markus Mohler Thomas C Wehler Carl C Schimanski department of Internal Medicine Marienhospital 64285 Darmstadt Germany Institute of Medical Biostatistics Epidemiology and Informatics Johannes Gutenberg University Hospital of Mainz 55131 Mainz Germany First department of Internal Medicine Johannes Gutenberg University Hospital of Mainz 55131 Mainz Germany department of Internal Medicine Katholische Kliniken Mainz 55131 Mainz Germany Institute of Pathology Saarland University Medical Center 66421 Homburg Germany Third department of Internal Medicine Johannes Gutenberg University Hospital of Mainz 55131 Mainz Germany

AIM: To analyze the efficacy of last line sorafenib treatment in colorectal cancer patients. METHODS: All patients receiving chemotherapy for colorectal cancer in the outpatient clinic of the University of Mainz since 2006 were retrospectively analyzed for last line sorafenib exposure. Charts of identified patients were analyzed for clinic-pathological parameters, like data on gender, age, date of initial diagnosis, UICC stage, number and kind of the pretherapies, therapy start and end of sorafenib, sorafenib mediated treatment cessation, side effects, response rates, time to progression and overall survival. RESULTS: Ten patients with a median of 3.0 prior chemotherapy lines had received a last line sorafenib therapy either alone(10%) or in combination with 5-fluorouracil derivates(90%). All patients suffered from colorectal cancer stage UICC 4 and were routinely seen in 2-wk intervals in the oncology outpatient clinic. Median duration of treatment was 142.0 d. At 8 wk 80% of patients showed stable disease but we did not observe any remissions. Median time to progression was 140.5 d(4.7 mo), while median overall survival reached 176.5 d. One patient ceased treatment due to side effects. Reason for treatment stop was bleeding complication in one case and non-specified sorafenib intolerance in another case. Due to the retrospective approach we did not further quantify side ***: This retrospective analysis encourages further investigation of sorafenib in colorectal cancer last line therapy.

关键词： Colorectal cancer Sorafenib Regorafenib Chemotherapy Last line

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Methylation status of the interferon-gamma gene promoter in chronic hepatitis B

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Journal of Pharmaceutical Analysis 2008年第3期20卷 206-212页

作者：陈瑞琳蔺淑梅叶峰张曦赵英仁张树林 department of Infectious Diseases the First Affiliated Hospital Medical School of Xi'an Jiaotong University

Objective To evaluate the methylation status at CpG site -55 in the interferon-gamma (IFN-γ) gene promoter and its effect on IFN-γ expression in chronic hepatitis B. Method The authors recruited 30 patients with HBeAg-positive chronic hepatitis B (CHB), 30 HBeAg-negative CHB patients, and 30 healthy blood donors. Pyrosequencing was used to determine the methylation status at CpG site -55 in the IFN-γ gene promoter following bisulfite treatment of DNA in peripheral blood mononuclear cells (PBMCs). The expression of IFN-γ was analyzed by real-time RT-PCR and ELISA. HBV DNA in PBMCs was detected by nested PCR. Results The methylation level at CpG site -55 in the IFN-γ gene promoter was significantly increased, resulting in subsequent down-regulation of the expression of this cytokine in CHB. The methylation level at CpG site -55 was significantly higher in HBeAg-positive patients than in HBeAg-negative ones (P<0.01) and was also significantly higher in PBMCs from HBV DNA-positive patients than from HBV DNA-negative ones (P<0.01); the methylation level at CpG site -55 was positively correlated with the amount of HBV DNA in serum (P<0.01). Conclusion IFN-γ gene expression appears to be regulated by methylation of the IFN-γ gene promoter in CHB; the methylation level at CpG site -55 is associated with HBV infection.

关键词： IFN-γ peripheral blood mononuclear cells(PBMCs) methylation hepatitis B virus CpG site

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Skin cancer in immunosuppressed transplant patients: Vigilance matters

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World Journal of Hepatology 2015年第4期7卷 717-720页

作者： Ozan Unlu Emir Charles Roach Alexis Okoh May Olayan Bulent Yilmaz Didem Uzunaslan Abdullah Shatnawei department of Pathobiology Cleveland Clinic Cleveland OH 44106 United States department of Gastroenterology Cleveland Clinic Cleveland OH 44106 United States department of Endocrine Surgery Cleveland Clinic Cleveland OH 44106 United States department of Internal Medicine Fairview Hospital Cleveland Clinic Cleveland OH 44106 United States department of Gastroenterology Hacettepe University School of Medicine 06230 Ankara Turkey

Liver transplantation(LT) is a widely-accepted, definitive therapy of irreversible liver diseases including hepatitis C, alcoholic liver disease and metabolic liver disease. After transplantation, patients generally use a variety of immunosuppressive medications for the rest of their lives to prevent rejection of transplanted liver. Mortality after LT is mainly caused by recurrence of alcoholic hepatitis which is mostly seen in the patients who resume heavy drinking. On the other hand, de-novo malignancies after LT are not seldom. Skin cancers make up 13.5% of the de-novo malignancies seen in these patients. Malignancies tend to affect survival earlier in the course with a 53% risk of death at 5 years after diagnosis. We aimed to report a case who underwent LT secondary to alcoholic liver disease and developed squamous cell carcinoma of the skin eighteen years after transplantation. In summary, transplant recipients are recommended to be educated on self examination for skin cancer; health care providers should be further suspicious during routine dermatological examinations of the transplant patients and biopsies of possible lesions for skin cancer is warranted even many years after transplantation.

关键词： Alcoholic liver disease Skin cancer Nonsquamous Liver transplantation Sirolimus

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Gastrointestinal tumors and infectious agents:A wide field to explore

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World Journal of Meta-Analysis 2021年第6期9卷 505-521页

作者： Miriam López-Gómez Belén García de Santiago Pedro-David Delgado-López Eduardo Malmierca Jesús González-Olmedo César Gómez-Raposo Carmen Sandoval Pilar Ruiz-Seco Nora Escribano Jorge Francisco Gómez-Cerezo Enrique Casado Medical Oncology department.Precision Oncology Laboratory Infanta Sofía University HospitalSan Sebastián de los Reyes 28231MadridSpain Pharmacy department Infanta Sofia University HospitalSan Sebastián de los Reyes 28703MadridSpain Neurosurgery department Burgos University HospitalBurgos 09006Spain Internal Medicine department Infanta Sofía University HospitalSan Sebastián de los Reyes 28703MadridSpain Medical Oncology department Infanta Sofia University HospitalSan Sebastián de los Reyes 28703MadridSpain Intensive Care Unit Jiménez Díaz FoundationMadrid 28040MadridSpain

Infection is currently one of the main contributors to *** fact,the International Agency for Research on Cancer has categorized eleven biological agents as group I *** is estimated that around 16%of the 12.7 million new cancers diagnosed in 2008 were attributable to infectious *** underdeveloped regions carry the highest incidence rates,about 7.4%of infectionrelated cancer cases occur in developed *** are increasingly aware of the potential carcinogenic role of common virus like the Human Papilloma virus in cervical cancer,or the hepatitis B and C viruses in ***,the carcinogenic role of several other infectious agents is less *** that gastrointestinal malignancies carry an overall poor prognosis,a better understanding of the carcinogenic mechanisms triggered by infectious agents is key to decrease the rate of cancer related *** measures directed to such infections would ideally impact *** this paper we review the main pathogenic mechanisms related to the development of gastrointestinal malignancies induced by infectious microorganisms and other pathogens which are currently under investigation.

关键词： Gastrointestinal tumors Infectious agents Bacteria Virus Prevention

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Systemic Inflammation in Patients with Chronic Obstructive Pulmonary Disease: Results from the Cosmic Study

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Open Journal of Respiratory Diseases 2012年第3期2卷 63-72页

作者： Marie-Kathrin Breyer Eric P. A. Rutten Martijn A. Spruit Wim C. J. Hop Dirkje S. Postma Emiel F. M. Wouters department of Biostatistics Erasmus Medical Center Rotterdam The Netherlands department of Pulmonology and Tuberculosis GRIAC Research Institute University Medical Center GroningenUniversity of Groningen Groningen The Netherlands department of Respiratory Medicine Maastricht University Medical Center (MUMC) Maastricht The Netherlands department of Respiratory Medicine Maastricht University Medical Center (MUMC) Maastricht The Netherlands Program Development Center Center of Expertise for Chronic Organ Failure Horn The Netherlands department of Respiratory Medicine Maastricht University Medical Center (MUMC) Maastricht The Netherlands Program Development Center Center of Expertise for Chronic Organ Failure Horn The NetherlandsDepartment of Respiratory & Critical Care Medicine & Ludwig Boltzmann Institute for COPD and Pulmonary Epidemiology Otto Wagner Hospital Vienna Austria

Objective: The study aims to elucidate the association of host-related factors on systemic inflammation in COPD patients. Methods: In 295 clinically stable and optimally treated COPD patients from 39 outpatient centers, age, gender, and body composition (body mass index, BMI;fat-free mass index, FFMI;fat mass index, FMI) were related to inflammatory biomarkers: CRP, fibrinogen, TNFα, and its soluble receptors (s)TNFαR1 and sTNFαR2. Furthermore, forced expiratory volume in the first second (FEV1), BMI, FFMI, and FMI were stratified by quartiles to elucidate the influence on inflammatory biomarkers. Monovariate and multivariate regression analyses were performed for associations between inflammatory biomarkers. Results: Positive correlations were found for FFMI with sTNFαR1, FMI with CRP and age with TNFα, sTNFαR1 and sTNFαR2 (p < 0.01). FEV1 was not correlated with body composition and inflammatory markers. Mono- and multivariate analysis showed weak correlations between the acute phase markers and the TNFα system after correcting for multiple co-variants. Conclusions: This study highlights the modest role of age and body composition on levels of systemic inflammatory biomarkers in COPD. Results show the degree of airflow limitation does not affect systemic inflammation. Last, a weak relationship between acute phase markers and markers of the TNFα system is present in COPD.

关键词： Adipokines Body Composition CRP Fibrinogen Obesity TNFα

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Dysfunction of invariant natural killer T cells in patients with Chronic Severe hepatitis B

Dysfunction of invariant natural killer T cells in patients ...

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第十一届全国免疫学学术大会

作者： Tan Xiaosheng Ding Yajie Dou Rui Hu Fen Zhang Lisha Yang Daofeng Liang Zhihui Wu Xiongwen Weng Xiufang department of Immunology Tongji Medical CollegeHuazhong University of Science and Technology department and Institute of Infectious Disease Tongji Hospital of Tongji Medical CollegeHuazhong University of Science and Technology

Background S: Chronic hepatitis B virus(HBV) infection is a major cause for liver injury which may progress to liver cirrhosis and hepatocellular carcinoma. Invariant natural killer T(iNKT) cells are innate-like lymphocytes abundant in liver tissue and mediate various functions. However, the reported function and role of iNKT cells in the disease progress after HBV infection are controversial. In this study, we examined functional status of iNKT cells from chronic severe hepatitis B(CSHB) patients, and aimed to elucidate the possible role of iNKT cells in the development of CSHB. Methods: 1) Ratio and phenotype of iNKT cells from peripheral blood of CSHB were tested by flow cytometry;2) Cytokine bead array were performed to determine their cytokine production capability;3) The responsiveness of iNKT cells from peripheral blood of CSHB was further determined by evaluating iNKT cell expansion upon α-Gal Cer stimulation in the presence of IL-2 and/or ***: iNKT cells were reduced significantly in peripheral blood of CSHB when compared to that of healthy donor, along with impaired cytokine production. The CD69 over-expression as well as higher levels of Fas L and Fas suggested iNKT cells from HBV infected patients suffered activation induced cell death. Further, residual activated iNKT cells from HBV infected patients defected in expansion upon α-Gal Cer stimulation, which could be partially recovered with IL-2 and/or IL-15. Conclution: iNKT cells were over-activated and exhibited dysfunctional cytokine production in CSHB patients, which may contribute to the immune disorders in the CSHB liver.

关键词： Chronic Severe hepatitis B(CSHB) iNKT cells Dysfunction AICD

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