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miR-34b/c regulates doxorubicin-induced myocardial cell injury through ITcH

cELL cYcLE 2019年第23期18卷 3263-3274页

作者： Zhang, Wen-cai Yang, Jin-Hua Liu, Guang-Hui Yang, Fan Gong, Jun-Long Jia, Meng-Ge Zhang, Meng-Juan Zhao, Luo-Sha Zhengzhou Univ Dept Cardiol Affiliated Hosp 1 Zhengzhou Henan Peoples R China

Objective: To determine the underlying mechanism of miR-34b/c in regulating doxorubicin (Dox)-induced myocardial cell injury. Methods: The viability of mouse myocardial cells HL-1 was detected by MTT assay. The apoptosis of HL-1 cells was detected by TUNEL assay. mRNA expressions of ITcH, TNF-? and IL-6 were measured by qRT-PcR. Protein levels of ITcH, NF-?B, TNF-? and IL-6 were measured by western blot. Dual luciferase assay was performed to detect the regulation of miR-34b/c on ITcH. Mouse model of cardiomyopathy was induced by intraperitoneal injection of Dox. Results: Dox reduced HL-1 cell viability and activated NF-?B pathway in HL-1 cells. miR-34b/c expressions were gradually up-regulated and ITcH expression was gradually down-regulated in Dox-treated HL-1 cells. miR-34b/c expression had negative correlation with the mRNA expression of ITcH. Besides, ITcH was a target of miR-34b/c. miR-34b/c mimic reduced cell viability, suppressed ITcH expression, increased TNF-? and IL-6 level, and promoted NF-?B expression in nucleus and cytoplasm of HL-1 cells. Whereas silencing miR-34 protected HL-1 cells through regulating ITcH. Finally, we demonstrated miR-34 antagomir-protected myocardial cells in mouse model of cardiomyopathy. conclusion: miR-34b/c decreased HL-1 cell viability and promoted the secretion of proinflammatory cytokines in Dox-induced myocardial cells through ITcH/NF-?B pathway.

关键词： Doxorubicin miR-34b c ITcH NF-?B myocardial cell

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Enhanced monocyte recruitment and delayed alternative macrophage polarization accompanies impaired repair following myocardial infarction in c57BL/6 compared to BALB/c mice

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cLINIcAL AND EXPERIMENTAL IMMUNOLOGY 2019年第1期198卷 83-93页

作者： Toor, I. S. Ruckerl, D. Mair, I. Thomson, A. Rossi, A. G. Newby, D. E. Allen, J. E. Gray, G. A. Univ Edinburgh Queens Med Res Inst BHF Univ Ctr Cardiovasc Sci 47 Little France Crescent Edinburgh EH16 4TJ Midlothian Scotland Univ Manchester Fac Biol Med & Hlth Sch Biol Sci Manchester Lancs England Univ Edinburgh Queens Med Res Inst MRC Ctr Inflammat Res Edinburgh Midlothian Scotland

Activation of the innate immune response following myocardial infarction (MI) is essential for infarct repair. Preclinical models of MI commonly use c57BL/6 mice, which have a type 1-dominant immune response, whereas other mouse strains such as BALB/c mice have a type 2-dominant immune response. We compared c57BL/6 and BALB/c mice to investigate whether predisposition towards a proinflammatory phenotype influences the dynamics of the innate immune response to MI and associated infarct healing and the risk of cardiac rupture. MI was induced by permanent coronary artery ligation in 12-15-week-old male wild-type BALB/c and c57BL/6 mice. Prior to MI, c57BL/6 mice had a lower proportion of cD206(+) anti-inflammatory macrophages in the heart and an expanded blood pool of proinflammatory Ly6c(high) monocytes in comparison to BALB/c mice. The systemic inflammatory response in c57BL/6 mice following MI was more pronounced, with greater peripheral blood Ly6c(high) monocytosis, splenic Ly6c(high) monocyte mobilization and myeloid cell infiltration of pericardial adipose tissue. This led to an increased and prolonged macrophage accumulation, as well as delayed transition towards anti-inflammatory macrophage polarization in the infarct zone and surrounding tissues of c57BL/6 mice. These findings accompanied a higher rate of mortality due to cardiac rupture in c57BL/6 mice compared with BALB/c mice. We conclude that lower post-MI survival of c57BL/6 mice over BALB/c mice is mediated in part by a more pronounced and prolonged inflammatory response. Outcomes in BALB/c mice highlight the therapeutic potential of modulating resolution of the innate immune response following MI for the benefit of successful infarct healing.

关键词： c57BL 6 BALB c macrophage polarization monocyte myocardial infarction

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Anti-hepatitis c virus therapy in chronic kidney disease patients improves long-term renal and patient survivals

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World Journal of clinical cases 2019年第11期7卷 1270-1281页

作者： Yi-chun chen chung-Yi Li Shiang-Jiun Tsai Yen-chun chen Division of Nephrology Department of Internal Medicine Dalin Tzu Chi Hospital Buddhist Tzu Chi Medical Foundation Chiayi County 622 Taiwan School of Medicine Tzu Chi University Hualien 970 Taiwan Department and Graduate Institute of Public Health College of Medicine National Cheng Kung University Tainan 701 Taiwan Department of Public Health College of Public Health China Medical University Taichung 404 Taiwan Department of Medical Research Dalin Tzu Chi Hospital Buddhist Tzu Chi Medical Foundation Chiayi County 622 Taiwan Division of Hepato-Gastroenterology Department of Internal Medicine Dalin Tzu Chi Hospital Buddhist Tzu Chi Medical Foundation Chiayi County 622 Taiwan

BAcKGROUND Hepatitis c virus (HcV) infection is a documented risk factor for chronic kidney disease (cKD) and progression to end-stage renal disease (ESRD). However, to date there are no reports on the long-term hard endpoints (ESRD and death) of anti-HcV therapy [interferon-based therapy (IBT) or new direct-acting antivirals] in cKD patients. Direct-acting antivirals are not available in Taiwan’s singlepayer national health insurance database currently released for research. Therefore, we hypothesized that a retrospective analysis of the long-term outcomes of IBT in cKD patients will serve as a proxy for direct-acting antivirals to increase our understanding of progression to ESRD following HcV infection. AIM To evaluate the long-term outcomes (ESRD and death) of anti-HcV therapy, especially IBT, in HcV-infected patients with stage 1-5 cKD. METHODS We analyzed 93894 Taiwan Residents adults diagnosed with cKD and without HBV infection. Of these, 4.9% were infected with HcV. Of the 4582 HcV-infected cKD patients, 482 (10.5%) received IBT (treated cohort). They were matched 1:4 with 1928 untreated HcV-infected cKD patients (untreated cohort) by propensity scores and year, which further matched 1:2 by propensity scores with 3856 cKD patients without HcV infection (uninfected cohort). All participants were followed until the occurrence of ESRD, death, or the end of 2012. The association between HcV infection, IBT use, and risks of ESRD and death was analyzed using competing risk analysis. RESULTS Taking the uninfected cohort as a reference, the adjusted hazard ratios for ESRD, after adjusting for competing mortality, were 0.34 (0.14-0.84, P = 0.019) and 1.28 (1.03-1.60, P = 0.029) in the treated and untreated cohorts, respectively. The treated cohort had a 29%(0.54-0.92, P = 0.011) decrease in mortality compared to the untreated cohort, in which the mortality was 31%(1.18-1.45, P < 0.001) higher than in the uninfected cohort. The reduced risks of ESRD (0.14, 0.03–0.58,

关键词： Hepatitis c virus chronic kidney disease End-stage renal disease Antihepatitis c virus therapy cohort study

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Predicting nitrogen mineralization from organic amendments: beyond c/N ratio by ¹³c-cPMAS NMR approach

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PLANT AND SOIL 2019年第1-2期441卷 129-146页

作者： Bonanomi, Giuliano Sarker, Tushar c. Zotti, Maurizio cesarano, Gaspare Allevato, Emilia Mazzoleni, Stefano Univ Naples Federico II Dipartimento Agr Via Univ 100 I-80055 Portici NA Italy Zhejiang A&F Univ Sch Environm & Resource Sci Hangzhou 311300 Zhejiang Peoples R China

Background and aim The processes of nitrogen (N) mineralization from organic amendments (OA) is important to provide mineral forms (NH4+ and NO3-) of N for plants. Assessment of N mineralization is usually done by using the c/N ratio of OAs but limits to the predicting capability of such indicator have been reported. Here, we propose a new definition of organic carbon (c) quality based on c-13-cPMAS NMR that provide a more accurate prediction of N mineralization. Methods A mesocosm incubation experiment was carried out to assess the effects of ten OA types on soil N mineralization in three different soils. OAs were chemically characterized by c/N ratio and in solid state spectroscopy by c-13-cPMAS NMR. Nitrogen mineralization in terms of NH4+ and NO3- was monitored after 3, 10, 30, 100, and 300 days of incubation. Results N mineralization was fast for OAs with high c quality coupled with high N content (e.g., meat powder, fish meal and alfalfa litter), while the same process was slow for amendment with low c quality even with high N content (e.g. humus, alfalfa biochar). On the contrary, a rapid but short-term N immobilization was found for OAs with high c quality but low N content (e.g., glucose). OAs with low c quality and low N content (e.g., sawdust, cellulose, wood biochar and grass litter) showed a slow, but long-lasting N immobilization. We found that c/N ratio was unable to predict the N mineralization for OAs with low c quality. considering c-13-cPMAS NMR spectral regions, the carbonyl c, and N-alkyl and methoxyl c regions had the most significant positive correlation with N mineralization, while the di-O-alkyl c and O-alkyl c were strongly associated with N immobilization. conclusions This study demonstrates that the biochemical quality of organic c defined by c-13-cPMAS NMR is capable of predicting N dynamic pattern better than c/N ratio.

关键词： N mineralization Organic amendment c N ratio c-13-cPMAS NMR Soil quality

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Exploring the hepatitis c virus genome using single molecule realtime sequencing

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World Journal of Gastroenterology 2019年第32期25卷 4661-4672页

作者： Haruhiko Takeda Taiki Yamashita Yoshihide Ueda Akihiro Sekine Department of Omics-based Medicine Center for Preventive Medical ScienceChiba UniversityChiba 260-0856Japan Department of Gastroenterology and Hepatology Graduate School of MedicineKyoto UniversityKyoto 606-8507Japan

Single molecular real-time(SMRT)sequencing,also called third-generation sequencing,is a novel sequencing technique capable of generating extremely long contiguous sequence *** conventional short-read sequencing cannot evaluate the linkage of nucleotide substitutions distant from one another,SMRT sequencing can directly demonstrate linkage of nucleotide changes over a span of more than 20 kbp,and thus can be applied to directly examine the haplotypes of viruses or bacteria whose genome structures are changing in real *** addition,an error correction method(circular consensus sequencing)has been established and repeated sequencing of a single-molecule DNA template can result in extremely high *** advantages of long read sequencing enable accurate determination of the haplotypes of individual viral *** sequencing has been applied in various studies of viral genomes including determination of the full-length contiguous genome sequence of hepatitis c virus(HcV),targeted deep sequencing of the HcV NS5A gene,and assessment of heterogeneity among viral ***,the emergence of multi-drug resistant HcV viruses has become a significant clinical issue and has been also demonstrated using SMRT *** this review,we introduce the novel third-generation PacBio RSII/Sequel systems,compare them with conventional next-generation sequencers,and summarize previous studies in which SMRT sequencing technology has been applied for HcV genome *** also refer to another long-read sequencing platform,nanopore sequencing technology,and discuss the advantages,limitations and future perspectives in using these thirdgeneration sequencers for HcV genome analysis.

关键词： Third generation sequencing PacBio RSII Single molecule real-time sequencing Hepatitis c virus Resistance-associated substitution Nanopore sequencer

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Peripheral cathepsin L inhibition induces fat loss in c. elegans and mice through promoting central serotonin synthesis

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BMc BIOLOGY 2019年第1期17卷 1-19页

作者： Lin, Yan Bao, Bin Yin, Hao Wang, Xin Feng, Airong Zhao, Lin Nie, Xianqi Yang, Nan Shi, Guo-Ping Liu, Jian Hefei Univ Technol Sch Food & Biol Engn 193 Tunxi Rd Hefei 230009 Anhui Peoples R China Univ Sci & Technol China Hefei Natl Lab Phys Sci Microscale Hefei 230026 Anhui Peoples R China Brigham & Womens Hosp Dept Med Boston MA 02115 USA Harvard Med Sch Boston MA 02115 USA Hefei Univ Technol Minist Educ Engn Res Ctr Bioproc Hefei 230009 Anhui Peoples R China

Background cathepsin L and some other cathepsins have been implicated in the development of obesity in humans and mice. The functional inactivation of the proteases reduces fat accumulation during mammalian adipocyte differentiation. However, beyond degrading extracellular matrix protein fibronectin, the molecular mechanisms by which cathepsins control fat accumulation remain unclear. We now provide evidence from caenorhabditis elegans and mouse models to suggest a conserved regulatory circuit in which peripheral cathepsin L inhibition lowers fat accumulation through promoting central serotonin synthesis. Results We established a c. elegans model of fat accumulation using dietary supplementation with glucose and palmitic acid. We found that nutrient supplementation elevated fat storage in c. elegans, and along with worm fat accumulation, an increase in the expression of cpl-1 was detected using real-time PcR and western blot. The functional inactivation of cpl-1 reduced fat storage in c. elegans through activating serotonin signaling. Further, knockdown of cpl-1 in the intestine and hypodermis promoted serotonin synthesis in worm ADF neurons and induced body fat loss in c. elegans via central serotonin signaling. We found a similar regulatory circuit in high-fat diet-fed mice. cathepsin L knockout promoted fat loss and central serotonin synthesis. Intraperitoneal injection of the cathepsin L inhibitor cLIK195 similarly reduced body weight gain and white adipose tissue (WAT) adipogenesis, while elevating brain serotonin level and WAT lipolysis and fatty acid beta-oxidation. These effects of inhibiting cathepsin L were abolished by intracranial injection of p-chlorophenylalanine, inhibitor of a rate-limiting enzyme for serotonin synthesis. conclusion This study reveals a previously undescribed molecular mechanism by which peripheral cPL-1/cathepsin L inhibition induces fat loss in c. elegans and mice through promoting central serotonin signaling.

关键词： Peripheral cathepsin L central serotonin Fat accumulation c elegans Mice

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Explaining trajectories of chemical changes during decomposition of tropical litter by ¹³c-cPMAS NMR, proximate and nutrients analysis

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PLANT AND SOIL 2019年第1-2期436卷 13-28页

作者： Sarker, Tushar c. Maisto, Giulia De Marco, Anna Esposito, Francesco Panico, Speranza c. Alam, M. Firoz Mazzoleni, Stefano Bonanomi, Giuliano Univ Naples Federico II Dept Agr Sci Via Univ 100 I-80055 Portici NA Italy Univ Naples Federico II Dept Biol Via Cinthia I-80126 Naples Italy Univ Rajshahi Dept Bot Rajshahi 6205 Bangladesh

Background and aimLitter decomposition is of great concern as it plays a key role in regulating global carbon cycle and nutrient budgets, especially in tropical forests where it is very fast. Therefore, we studied the decomposition and subsequent changes in element concentrations and amounts of 11 tropical leaf *** litters were characterized in terms of elemental, proximate features, as well as organic carbon compositions by c-13-NMR spectroscopy. controlled litterbag decomposition was carried out for 180days in the laboratory, bags were retrieved at three dates and analyzed for mass loss and concentration of nitrogen (N), potassium (K), sodium (Na), carbon (c), magnesium (Mg), manganese (Mn), iron (Fe), phosphorus (P), zinc (Zn), and percentage of ash, extractives, cellulose and *** arjuna showed highest mass loss, while the lowest was observed in Shorea robusta showing significant positive correlation with litter initial K, Mg, Fe and di-O-alkyl c, O-alkyl c, while significant negative correlation with Mn, lignin and Methoxyl c, Alkyl c. Trajectories shows that Fe, Mg, K, Mn, Zn and Na concentrations increased in most of the litter types, except Terminalia arjuna and Toona ciliata, where Fe, Mg, K concentrations decreased during decomposition. Surprisingly, in most of the species with the exception of Terminalia arjuna, N concentrations decreased at a faster rate than c concentrations, resulting in an increase in c/N ratios. Expectedly, lignin progressively increased, while cellulose *** differences in decomposition rate and dynamics of chemical changes among litters were strongly affected by the litter type, decomposition time and their interactions.

关键词： Tropical litter Nutrients dynamics Proximate features c N ratio Decomposition

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Blocking c/EBP β protects vascular endothelial cells from injury induced by intermittent hypoxia

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SLEEP AND BREATHING 2019年第3期23卷 953-962页

作者： Feng, Yu Li, Qingchu Wu, Yinxiang Zhao, Nana Li, Lu Li, Li Zhao, Liming Tongji Univ Shanghai East Hosp Dept Resp Med Sch Med Shanghai 200120 Peoples R China Second Mil Med Univ Dept Radiol Secondary Affiliated Hosp Shanghai 200001 Peoples R China Second Mil Med Univ Dept Resp Med Affiliated Hosp 3 Shanghai 200438 Peoples R China Second Mil Med Univ Dept Teaching Support Shanghai 200433 Peoples R China

Background Intermittent hypoxia (IH) can damage endothelial cells and lead to apoptosis in obstructive sleep apnea-hypopnea syndrome (OSAHS). Hypoxia induces apoptosis in endothelial cells via upregulation of endothelin-1 (ET-1) and hypoxia inducible factor-1 alpha (HIF-1 alpha) plays a key role in the hypoxic stress response. Purpose We investigated an approach to diminish the negative effect of HIF-1 alpha while maintaining its protective effect. Methods Human umbilical vein endothelial cells (HUVEcs) were subjected to sustained hypoxia (SH) or IH for 24 h, and the responses of HIF-1 alpha, ccAAT/enhancer binding protein beta (c/EBP beta), and endothelin-1 (ET-1) were assessed by western blotting. A luciferase reporter system was employed to verify the potential binding site (transcription factor binding site, TFBS) for c/EBP beta in the ET-1 promoter. The specificity of regulation of ET-1 by HIF-1 alpha via c/EBP beta was evaluated by a lentiviral system. The effects of silencing of c/EBP beta on IH-induced apoptosis, vascular endothelial growth factor (VEGF) protein levels, proliferation, and in vitro tube formation were studied. Results We found that IH significantly increased HIF-1 alpha, c/EBP beta, and ET-1 in HUVEcs. Knockdown of HIF-1 alpha or c/EBP beta inhibited the upregulation of ET-1 induced by IH. Blocking c/EBP beta impaired IH-induced apoptosis but did not affect VEGF expression, proliferation, or in vitro tube formation. c/EBP beta was shown to mediate increased ET-1 transcription by HIF-1 alpha through the TFBS, 5 '-GTTGccTGTTG-3 ', in ET-1 promoter. conclusion Silencing of c/EBP beta can suppress apoptosis but does not affect the protective role of HIF-1 alpha in the hypoxic stress response.

关键词： OSAHS Intermittent hypoxia HIF-1 alpha ET-1 c EBP beta

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Diffusion-weighted magnetic resonance imaging and micro-RNA in the diagnosis of hepatic fibrosis in chronic hepatitis c virus

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World Journal of Gastroenterology 2019年第11期25卷 1366-1377页

作者： Tarek Besheer Hatem Elalfy Mohamed Abd El-Maksoud Ahmed Abd El-Razek Saher Taman Khaled Zalata Wagdy Elkashef Hossam Zaghloul Heba Elshahawy Doaa Raafat Wafaa Elemshaty Eman Elsayed Abdel-Hady El-Gilany Mahmoud El-Bendary Department of Tropical Medicine and Hepatology Mansoura Faculty of Medicine-Mansoura University Department of Diagnostic Radiology Mansoura Faculty of Medicine-Mansoura University Department of Pathology Mansoura Faculty of Medicine-Mansoura University Department of Clinical Pathology Mansoura Faculty of Medicine-Mansoura University Department of Public Health and Preventive Medicine Mansoura Faculty of Medicine-Mansoura University

BAcKGROUND Diffusion-weighted magnetic resonance imaging has shown promise in the detection and quantification of hepatic fibrosis. In addition, the liver has numerous endogenous micro-RNAs(miRs) that play important roles in the regulation of biological processes such as cell proliferation and hepatic *** To assess diffusion-weighted magnetic resonance imaging and miRs in diagnosing and staging hepatic fibrosis in patients with chronic hepatitis *** This prospective study included 208 patients and 82 age-and sex-matched controls who underwent diffusion-weighted magnetic resonance imaging of the abdomen, miR profiling, and liver biopsy. Pathological scoring was classified according to the METAVIR scoring system. The apparent diffusion coefficient (ADc) and miR were calculated and correlated with pathological *** The ADc value decreased significantly with the progression of fibrosis, from controls(F0) to patients with early fibrosis(F1 and F2) to those with late fibrosis(F3 and F4)(median 1.92, 1.53, and 1.25 × 10^(-3) mm^2/s, respectively)(P = 0.001).The cut-off ADc value used to differentiate patients from controls was 1.83 × 10^(-3) mm^2/s with an area under the curve(AUc) of 0.992. combining ADc and miR-200 b revealed the highest AUc(0.995) for differentiating patients from controls with an accuracy of 96.9%. The cut-off ADc used to differentiate early fibrosis from late fibrosis was 1.54 × 10^(-3) mm^2/s with an AUc of 0.866. The combination of ADc and miR-200 b revealed the best AUc(0.925) for differentiating early fibrosis from late fibrosis with an accuracy of 80.2%. The ADc correlated with miR-200 b(r =-0.61, P = 0.001), miR-21(r =-0.62, P = 0.001), and miR-29(r = 0.52,P = 0.001).cONcLUSION combining ADc and miRs offers an alternative surrogate non-invasive diagnostic tool for diagnosing and staging hepatic fibrosis in patients with chronic hepatitis c.

关键词： Diffusion Magnetic resonance imaging Fibrosis Liver Hepatitis c virus Micro-RNA

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Hepatitis c virus clearance and less liver damage in patients with high cholesterol, low-density lipoprotein cholesterol and APOE ε4 allele

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World Journal of Gastroenterology 2019年第38期25卷 5826-5837页

作者： Karina Gonzalez-Aldaco Sonia Roman Rafael Torres-Valadez claudia Ojeda-Granados Luis A Torres-Reyes Arturo Panduro Department of Molecular Biology in Medicine Civil Hospital of Guadalajara "Fray Antonio Alcalde" and Health Sciences Center University of Guadalajara

BAcKGROUND cholesterol is related to improvements in the rate of sustained virological response and a robust immune response against the hepatitis c virus(HcV).APOE gene polymorphisms regulate cholesterol levels modifying the course of the HcV *** relationship between cholesterol,APOE alleles,and the outcome of HcV infection has not been evaluated in the admixed population of *** To investigate the role of APOE-ε2,-ε3,and-ε4 alleles and the metabolic profile in the outcome of HcV *** A total of 299 treatment-na?ve HcV patients were included in this retrospective *** were stratified in chronic hepatitis c(cHc)(n=206)and spontaneous clearance(Sc)(n=93).A clinical record was *** tests were assessed by dry chemistry *** genotypes were determined using a Real-Time polymerase chain reaction *** Total cholesterol,low-density lipoprotein cholesterol(LDL-c),triglycerides,and hypercholesterolemia were higher in Sc than cHc patients as well as the frequency of the APOEε4 allele(12.4%vs 7.3%).Sc patients were overweight(54.8%).Theε4 allele was associated with Sc(OR=0.55,95%cI:0.31-0.98,P=0.042)and mild fibrosis(F1-F2)in cHc patients(OR 0.091,95%cI 0.01-0.75,P=0.020).LDL-c≥101.5 mg/dL(OR=0.20,95%cI:0.10-0.41,P<0.001)and BMI≥26.6 kg/m2(OR=0.37,95%cI:0.18-0.76,P<0.001)were associated with Sc status;while ALT≥50.5 IU/L was negatively associated(OR=5.67,95%cI:2.69-11.97,P<0.001).cONcLUSION In Sc patients,the APOEε4 allele and LDL-c conferred a protective effect in the course of the HcV infection in the context of excess body weight.

关键词： Liver damage Body mass index Spontaneous hepatitis c virus clearance Low-density lipoprotein cholesterol

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