Comparison of Hepatocellular Carcinoma Patients With Alcoholic Liver Disease and Nonalcoholic Steatohepatitis

Background: Alcoholic hepatitis and nonalcoholic steatohepatitis (NASH) show different clinical features with similar liver histology, but both disorders may progress to cirrhosis and hepatocellular carcinoma (HCC). HCC arising in alcoholic liver disease (ALD) or NASH, without hepatitis B or C virus infection, has been a rare observation, and there are no studies comparing the characteristics of ALD and NASH patients with HCC. Therefore, we compared the characteristics of ALD and NASH patients with HCC.
Methods: A total of 1202 patients received a diagnosis of HCC at Tokyo Women's Medical University from 1989 to 2003, and their clinical data were collected prospectively. A clinical diagnosis was made to diagnose ALD, and clinical and histological changes were required to diagnose NASH. Of these patients, 88 received a diagnosis of HCC arising from ALD. Among them, a biopsy specimen was obtained in 50 patients (ALD-HCC group). We compared the clinical and histological characteristics of 50 ALD and 8 NASH patients (NASH-HCC group) associated with HCC. They all were negative for hepatitis virus infection by serological methods.
Results: The most significant difference between these groups was sex. Women were significantly more common in the NASH-HCC group (6% vs. 63%; p < 0.0001). The median age was 65 years in the ALD-HCC group and 68 years in the NASH-HCC group. The risk factors for NASH all were high in the NASH-HCC group. However, liver function tests were similar in these groups. In the ALD-HCC group, 46 (92%) patients showed severe fibrosis; 2 had septal fibrosis and 44 had cirrhosis. All patients in the NASH-HCC group showed severe fibrosis, and seven had cirrhosis.
Conclusions: Severe fibrosis might be an important risk factor for HCC. Patients who have ALD or NASH with cirrhosis may develop HCC. This seems to occur in a sufficient number of cases to warrant regular screening for this complication.     

Mutation of Mitochondrial DNA in Livers From Patients With Alcoholic Hepatitis and Nonalcoholic Steatohepatitis

Background: In alcoholic hepatitis (Al-Hep) and nonalcoholic steatohepatitis (NASH), triglycerides accumulate in hepatocytes. We examined the hypothesis that mutations in mitochondrial DNA may take place by mitochondrial overwork, resulting in dysfunction of mitochondria.
Subjects and Methods: Subjects of this research were 8 cases each of Al-Hep, NASH, and fatty liver (FL). Total DNA was extracted from the biopsied liver samples. DNA fragments were amplified by PCR and DNA sequences determined in the control and coding regions of mitochondrion.
Results: When the numbers of mutations per 1,000 bases of mitochondrial DNA were compared between each group, no significant differences were found among D-loop, HV1, and HV2 mitochondrial DNA regions. However, there were significantly more mutations in ND1 and COII of Al-Hep and NASH than in FL, and mutations were comparatively at random. Neither a region in which mutations were focused nor differences among the groups were recognized. When details of the base mutation in a control region were investigated by group, the transition type of mutation between T:A≪–≫C:G occurred in at least 70%. Also, a transition-type mutation was found mostly in a coding region, which was similar to the mutation pattern in the control region, except for the ND1 and COII regions where there were hardly any mutations.
Conclusions: As gene mutations of mitochondrial DNA appeared frequently in Al-Hep, and also in NASH, mitochondrial dysfunction caused by mutation in mitochondrial DNA may be involved in the pathogenesis of both diseases.     

Comparison between Nonalcoholic Steatohepatitis and Alcoholic Hepatitis

Two groups, 16 nonalcoholic steatohepatitis patients (group I) and 22 alcoholic hepatitis patients (group II) classified according to the presence or absence of drinking and their histological characteristics, were compared on the basis of clinical, biochemical, and liver biopsy findings. The frequencies of female patients (p less than 0.01), obesity (p less than 0.001), and maturity-onset diabetes (p less than 0.005) were significantly greater in group I than in group II. The serum glutamic pyruvic transminase (p less than 0.05) and gamma-glutamyltranspeptidase (p less than 0.05) contents were significantly greater in group II than in group I. The cholinesterase content (p less than 0.05) was significantly less in group II. Significant differences were found in the grades of nuclear vacuolation (p less than 0.001, Fisher's exact probability test), periportal pericellular fibrosis, proliferation of bile ductules, and changes in the shape of the portal tracts (p less than 0.001, Wilcoxon's rank-sum test). Zonal necrosis in group I was seen in only severe steatohepatitis. These clinical and biochemical findings were found to be useful in differentiating nonalcoholic steatohepatitis from alcoholic hepatitis. Liver biopsy was of limited value at best in separating the two conditions.     

Independent Predictors of Steatohepatitis and Fibrosis in Asian Indian Patients with Non-Alcoholic Steatohepatitis

AIM: To identify the clinical and biochemical predictors of histologic disease severity in patients with non-alcoholic steatohepatitis (NASH). METHOD: Clinical, anthropometric and biochemical data of 71 consecutive patients with a histological diagnosis of NASH were retrieved from the medical database. The histologic criteria proposed at the American Association for the Study of Liver Diseases Conference (2003) were followed. Histologic features and the necro-inflammatory grade of liver biopsy were scored in accordance with the Brunt score. Statistical analysis was performed to identify predictors of disease severity. RESULTS: The male to female ratio of the study cohort was 3.1:1. The mean body mass index (BMI) of the patients was 25.67 +/- 1.9 kg/m2. In terms of necroinflammatory grading, 21 (29.6%) patients were classified as grade 1, 26 (36.6%) as grade 2 and 24 (33.8%) as grade 3. Multivariate analysis revealed that BMI (P = 0.009), waist circumference (P = 0.035), waist:hip ratio (P = 0.011) and aspartate aminotransferase levels (P < 0.001) were independent predictors of necroinflammatory grade and that female gender (P = 0.02), serum alkaline phosphatase levels (P = 0.018), cholesterol levels (P = 0.048) and low-density lipoprotein (LDL) levels (P = 0.025) were independent predictors of fibrosis stage. CONCLUSION: Female gender, BMI, waist:hip ratio, hypercholesterolemia and LDL levels are independent predictors of disease severity in patients with NASH and may influence the decision to biopsy.     

Immunomodulatory effects of feeding with Bifidobacterium longum on allergen-induced lung inflammation in the mouse

BackgroundThe intestinal microbiota has important effects on host immune responses and feeding with certain commensal organisms has anti-inflammatory effects in a variety of diseases, including experimental asthma. The aim of the current study was to examine how robust the effects of feeding with the commensal strain, Bifidobacterium longum (Bif) were on the pulmonary responses to allergen sensitization and challenge.MethodsBALB/c mice were given two intraperitoneal injections of ovalbumin (10 μg in alum) on days 0 and 7 and were fed daily with Bif or vehicle from days 0–14. Challenges with ovalbumin (10 μg) were administered intra-nasally once on day 14 or three times on days 14, 15 and 16 and the lung inflammatory response was assessed one day later.ResultsBif feeding attenuated airway inflammation following a single ovalbumin challenge, reducing bronchoalveolar lavage (BAL) eosinophilia, BAL fluid IL-4 protein and BAL cell IL-4 and IFN-γ mRNA levels. However, BAL fluid IL-5 protein was increased. There was an accompanying increase in lung regulatory T cells assessed by flow cytometry. Responses to triple challenge with ovalbumin were much less affected by Bif feeding, including unchanged cytokine levels, ovalbumin-specific IgE and airway hyperresponsiveness to methacholine.ConclusionThese results show modest immunoregulatory effects of oral feeding with Bif with inhibition of certain components of allergen-induced airway inflammation that is associated with the expansion of regulatory T cells in the lungs but that is overcome by repeated allergen exposure.     

Predictors of Cirrhosis in Hispanic Patients with Nonalcoholic Steatohepatitis

It is estimated that 43% of patients with nonalcoholic steatohepatitis (NASH) will progress to liver fibrosis or cirrhosis. Although NASH is more common in Hispanics, most studies have been conducted on Caucasians, and there is scarce information regarding ethnic differences in this disease. The aim of this study was to identify the independent predictors of cirrhosis in Hispanic patients with NASH. A retrospective case-control study was conducted on 80 patients with biopsy-proven NASH. Forty-two were Hispanic (study group) and 38 were Caucasians controlled for age and BMI (control group). Clinical, biochemical, and histologic features were analyzed for correlation with cirrhosis. There were no significant differences in demographic features between the two groups. In multivariate analysis, independent predictors of cirrhosis among Hispanic patients were age (OR, 1.07; 95% CI, 1.01–1.14) and AST/ALT ratio (OR, 10.56; 95% CI; 2.46–45.29), while independent predictors among non-Hispanic patients were age (OR, 1.085; 95% CI, 1.0–1.186), and diabetes mellitus (OR, 6.46; 95% CI, 1.19–35.07). In patients with NASH, predictors of cirrhosis varied according to ethnic background. Age was an independent predictor in both groups, however, AST/ALT ratio was found to be an independent predictor of cirrhosis only in Hispanic patients, and diabetes mellitus only in non-Hispanic patients.     

Association of Apolipoprotein E Polymorphisms in Patients with Non-Alcoholic Steatohepatitis

Non-alcoholic fatty liver disease (NAFLD) is the most common cause of abnormal hepatic steatosis in the absence of alcohol abuse worldwide. Non-alcoholic steatohepatitis (NASH) is the most progressive form of NAFLD. The aim of this study was to investigate the role of apolipoprotein E (APOE) polymorphisms in the development of NASH. We analysed 57 NASH patients and 245 healthy controls using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in a case–control study. The diagnosis of the patients was based on liver biopsy. The serum levels of glucose, lipids, vitamin B12, folic acid, homocysteine, insulin, total biluribin, total protein, albumin, ferritin, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were determined in all of the subjects. Body mass index (BMI), waist circumference (WC), AST, ALT, fasting blood sugar (FBS), total cholesterol, triglyceride (TG), low-density lipoprotein (LDL) cholesterol, very low-density lipoprotein (VLDL) cholesterol, insulin and ferritin levels were significantly higher in the 57 patients with NASH compared with the 245 healthy controls. The APOE ε3 allele was overrepresented in the whole group of NASH patients (ε3=97.37% in NASH versus 82.45% in controls). The APOE polymorphism was statistically significantly associated with NASH (χ²=15.741; p=0.008). The APOE3/3 genotype (odds ratio [OR]=7.941; p=0.000) was strongly associated with increased risk for NASH in all NASH patients. Consequently, the APOE3/3 genotype may play a role in the aetiopathogenesis of NASH.     

CFTR Polymorphisms in Patients with Alcoholic Chronic Pancreatitis

Introduction: Pancreas susceptibility to alcohol is variable and only 5–10% of chronic alcohol abusers develop chronic pancreatitis; the role of genetic factors in this process is unknown. The CFTR gene encodes a protein that acts on epithelial cells and plays a key role in normal exocrine pancreatic function. Methods: This study investigated the frequency of polymorphisms in intron 8 of the CFTR gene in patients with alcoholic chronic pancreatitis. Three groups of patients were studied: group A - 68 adult alcoholics with a diagnosis of chronic pancreatitis; group B - 68 adult alcoholics without pancreatic disease or liver cirrhosis and group C - 104 healthy nonalcoholic adults. Results: T5/T7 genotype was more frequent in group A (11.8%) than in group B (2.9%) (p = 0.0481), and there was no statistical difference when groups A and C (5.8%) were compared (p = 0.1317). The haplotype combination (TG)10-T7/(TG)11-T7 was more frequent in groups B (23.5%) and C (20.2%) than in group A (7.3%) (p = 0.0080 and 0.0162). Conclusion: There are differences when these three groups are compared and individuals with T5/T7 genotype might have a greater risk of developing chronic pancreatitis when they become chronic alcoholics.     

猪带绦虫TSO45W-4B-TSOL18融合基因在长双歧杆菌中的表达

目的 在成功构建猪带绦虫大肠杆菌-双歧杆菌穿梭表达质粒pGEX-TSO45W-4B-TSOL18的基础上,研究猪带绦虫TSO45W-4B-TSOL18融合基因在长双歧杆菌中的表达情况。方法 将猪带绦虫大肠杆菌-双歧杆菌穿梭表达质粒pGEX-TSO45W-4B-TSOL18电转化入长双歧杆菌, IPTG诱导表达,SDS-PAGE和Western blot分析表达情况。结果 酶切、PCR和测序证实,重组质粒pGEX-TSO45W-4B-TSOL18成功转入长双歧杆菌。SDS-PAGE显示,重组蛋白相对分子质量(M_r)约为55 kD,与预期结果相一致。Western blot显示,重组蛋白能被兔抗血清、囊虫病猪血清和囊虫病患者血清所识别。结论 猪带绦虫TSO45W-4B-TSOL18融合基因能够在长双歧杆菌中获得表达,表达的重组蛋白具有特异的抗原性。     

长双歧杆菌抑制回肠NLRP6炎症小体调节感染后肠易激综合征内脏高敏感性

[目的]研究NLRP6炎症小体在长双歧杆菌调节感染后肠易激综合征(PI-IBS)内脏高敏感性中的作用。[方法]NIH小鼠40只分为5组(每组8只):2周正常对照组(NC-2W组),感染后2周组(PI-2W组),8周正常对照组(NC-8W组),感染后8周组(PI-8W组)及感染后8周长双歧杆菌干预组(PI-B组);腹壁回撤反射(AWR)检测各组内脏敏感性,Western-blot检测CD172a以评价肠道炎症程度,Western-blot检测各组小鼠末端回肠NLRP6及其配体ASC、caspase-1、CARD8以及下游细胞因子白细胞介素(IL)-18、IL-1β表达。[结果]1PI-2W组和PI-8W组内脏敏感性较NC-2W组和NC-8W组明显增加(P0.05),PI-B组内脏敏感性较PI-8W组明显降低(P0.05);2PI-2W组与NC-2W组比较,CD172a表达水平显著升高(P0.01);PI-8W组与NC-8W组比较,CD172a表达水平差异无统计学意义(P0.05);PI-B组与PI-8W组比较,CD172a表达水平差异无统计学意义(P0.05);3 PI-2W组和PI-8W组NLRP6、CARD8、ASC和caspase-1及IL-18均较NC-2W组和NC-8W组明显增加(P0.05),而PI-B组NLRP6、CARD8、ASC和caspase-1及IL-18较PI-8W组明显降低(P0.05),PI-2W组IL-1β表达水平较NC-2W组显著升高(P0.05),而NC-2W组、NC-8W组、PI-8W组、PI-B组间IL-1β表达水平比较均差异无统计学意义(P0.05)。[结论]长双歧杆菌可能通过抑制炎症小体NLRP6,下调IL-18,从而降低PI-IBS内脏高敏感性。     

Effect of Bifidobacterium longum 35624 on disease severity and quality of life in patients with irritable bowel syndrome

BACKGROUND Bifidobacterium longum 35624 has shown efficacy in improving irritable bowel syndrome(IBS)symptoms compared with placebo in double-blind randomized studies.However,few data are available from real-life clinical practice or from studies that used Rome IV criteria to diagnose IBS.AIM To assess the effect of B.longum 35624 on IBS severity and quality of life in a reallife setting.METHODS From November 2018 to January 2020,278 patients with IBS(according to Rome IV criteria)were enrolled in a prospective,open-label,multicenter observational study by private practice gastroenterologists to received one capsule of B.longum 35624(10⁹ colony-forming units)per day for 30 d.Participation in the study was independently proposed to patients during spontaneous consultations.Disease severity(assessed by the IBS severity scoring system)and patient quality of life(assessed by the IBS quality of life questionnaire)were compared between the inclusion visit(baseline)and the visit at the end of 30 d of treatment.The characteristics of patients were described at baseline.Continuous variables comparisons between inclusion and end-of-treatment visits were performed using the t-test and Kruskal-Wallis test.Categorical variables comparisons were performed using theχ² test.RESULTSA total of 233 patients,with a mean age of 51.4 years and composed of 71.2%women,were included in the study.Of these patients,48.1%had moderate IBS and 46.4%had severe IBS.After a 30-d treatment period with one B.longum 35624 capsule per day,a significant decrease in IBS severity was observed compared to baseline(mean±SD,IBS severity scoring system scores:208±104 vs 303±81,P<0.001)and 57%of patients moved to lower severity categories or achieved remission.The quality of life of patients was also improved by the treatment(IBS Quality of Life questionnaire score:68.8±20.9 vs 60.2±20.5;P<0.001)and 63.8%of patients were satisfied with the treatment.CONCLUSION Thirty days of treatment with B.longum 35624 reduces disease severity and improves the quality of life of patients with IBS,particularly those with the most severe forms of IBS.