Adult ; Female ; Humans ; Liver ; pathology ; Male ; Middle Aged ; Non-alcoholic Fatty Liver Disease ; pathology ; Obesity ; pathology ; Young Adult

Objective To evaluate the role of two-dimensional ultrasound combined with contrast-enhanced ultrasonography (CEUS) in the classification of liver nodules in cirrhotic patients.Methods Consecutively cirrhotic patients with intrahepatic nodules at Xixi Hospital of Hangzhou were included from November 2015 to December 2016.All (142 nodules in 109 patiens) presented as non-cancerous focal lesions on conventional magnetic resonance imaging and CT examination and had available information of liver biopsy.Each lesion was percutaneous biopsied under the guidance of two-dimensional ultrasound.Ultrasonographic parameters evaluated were as following:(1) sizes of nodules under US;(2) ultrasonographic characteristics of the nodular;(3) CEUS enhancement features of the nodules.Four types of hepatic nodule suggesting different histology were defined according to the ultrasonographicparameters.x2 test was used to compare the difference of hepatocellular carcinoma (HCC) incidence among liver nodules with varying sizes and nodules with different enhancement features under CEUS.As for the statistical differences of HCC and high-grade dysplastic nodule (HGDN) incidence between type Ⅲ & Ⅳ nodules and type Ⅰ & Ⅱ nodules,x2 test was also used for analysis.Results A total of 142 eligible nodules were detected in 109 patients with cirrhosis,including 16 HCCs,2 intrahepatic cholangiocellular carcinomas (ICC),41 HGDNs,40 low-grade dysplastic nodules (LGDN) and 43 regenerative nodules (RN).In terms of diameter,all (6/6) the nodules larger than 2.0 cm,20.0％ (8/40) of middle size nodules (1.5-2.0 cm),were HCCs.The remained 2 lesions of HCC came from two subgroups with even small size nodules [1.0-1.4 cm (n=93),and ＜ 1.0 cm (n=3),in diameter],respectively.Two lesions of ICC were attributed to nodules with a 1.0-1.4 cm diameter.About 28 nodules with a diameter of 1.5-2.0 cm,13 nodules with a diameter of 1.0-1.4 cm were HGDN.HCC incidences between these 4 groups were different significantly (x2=61.425,P ＜ 0.001).Asfor the CEUS,14 nodules exhibited a rapid enhancement feature in arterial phase,12 of which were HCC.In56 nodules with a slow enhancement feature,4 nodules were HCC.HCC incidences between these 3 groups were different significantly (x2=75.752,P ＜ 0.001).Under the combined ultrasonography,HCC incidences of type Ⅲ and type Ⅳ nodules were significantly higher than that of type Ⅰ and type Ⅱ lesions [21.9％ (16/73)vs 0 (0/65),x2=15.222,P ＜ 0.001],similar result was observed in the comparison of HGDN incidences between type Ⅲ & Ⅳ and type Ⅰ & Ⅱ nodules[53.4％ (39/73) vs 3.1％ (2/65),x2=38.842,P ＜ 0.001].Conclusion The classification presented by this study,combining the three ultrasonographic parameters,which is nodule size,nodular echo characteristics and enhancement features of the nodules under CEUS,could be helpful for the diagnosis of HCC in cirrhotic patients with ill-defined nodule on routine image examination.

Objective To evaluate the prevalence of hepatic steatosis in patients with chronic hepatitis B (CHB) and the impact of hepatic steatosis on the progress of fibrosis. Methods Five hundred and sixty two untreated CHB patients (405 males and 157 females) with an average age of 31.3 underwent liver biopsy from January to August 2007. On the day of liver biopsy, a questionnaire was completed and a blood sample was obtained for laboratory analysis. The degree of liver steatosis, necroinflammation and fibrosis was assessed; demographic information and clinical data including age, gender, body mass index (BMI), history of diabetes mellitus, hypertension, dyslipidemia, and HBeAg status, HBV DNA viral load were documented. Results In 562 patients, 102 (18. 2% ) had steatosis. Multivariate analysis demonstrated that liver steatosis was associated with the levels of TG, APO-B, UA, FSG, and higher BMI; and the progress of fibrosis was associated with high degree of hepatic steatosis and necroinflammation, age over 35 years, HBV DNA > 103 copies/L, high BMI and GGT. Conclusions The results show that obesity and dyslipidemia in CHB patients are associated with the hepatic steatosis, and the latter seems to be an important determinant for fibrosis.

The traditional Chinese medicine (TCM) constitution theory is an important means to reveal the host′s genetic characteristics from the perspective of TCM. The types of constitution can influence liver pathological changes and the prognosis of disease in patients with chronic hepatitis B virus infection, and to a certain degree, they are associated with the polymorphisms of genes involved in immunoregulation, for example, human leukocyte antigen(HLA) class Ⅱ gene. Yin-deficiency constitution is associated with various adverse clinical outcomes, as well as the genotypes of genes including HLA-DQA1*0501. Preliminary data also show that the chronic hepatitis B (CHB) patients with yin-deficiency constitution have a poor response to interferon therapy. This article reviews the current application of TCM constitution theory in the diagnosis and treatment of CHB, its potential value, and existing problems.

Objective: To investigate the clinical features and reliable diagnostic method in HIV/AIDS patients with smear negative pulmonary tuberculosis (TB). Methods: Clinical data of 112 HIV/AIDS patients complicated with smear negative pulmonary TB who were treated in our hospital from January 2013 to September 2015 were retrospectively analyzed. These clinical data includeded clinical symptom, blood routine test, blood biochemistry, T lymphocyte subsets classification, sputum acid-fast bacillus smear, mycobacterium tuberculosis culture, purified protein derivatives tuberculin (PPD) test, interferon gamma-release assay for Mycobacterium tuberculosis (T-SPOT.TB), TB-DNA and chest computed tomography (CT). Diagnostic specificity and sensitivity of these parameters were analyzed. Results: No specific clinical manifestation of these patients was identified. The chest CT feature was also atypical. The positive rates including T-SPOT, TB, TB-DNA and PPD test were all low. The positive rates of T-SPOT.TB and PPD test in patients with a CD4+ cell count >200 cells/μl was significantly higher than that of patients with a CD4+ cell count ≤50 cells/μl and 51≤CD4≤200 cells/μl (P<0.01). Conclusions The clinical feature of smear negative pulmonary TB in HIV/AIDS patients is atypical. For a definite diagnosis, a comprehensive analysis with clinical manifestations, laboratory test, imaging examination and etiologic detection is required.

Objective:To conduct clinical and genetic analysis in two cases of cholestatic liver disease to determine the specific etiology of cholestasis.Methods:Clinical data and the medical histories in family members of two cases were collected. The gene variation was detected by whole-exome sequencing technology. Sanger sequencing validation and bioinformatics analysis were performed on patients and their parents with suspected pathogenic mutations.Results:Whole-exome sequencing showed that the ABCB4 gene of case 1 (a male, 16 years old) had compound heterozygous mutations of c.646C > T from the father and c.927T > A from the mother, while the ABCB4 gene of case 2 (a female, 17 years old) had a compound heterozygous mutation of c.2784-1G > A from the father and c.646C > T from the mother. New mutation sites that had not been previously reported were c.646C > T, c.927T > A, and c.2784-1G > A.Conclusion:In this study, both cases had progressive familial intrahepatic cholestasis type 3 (PFIC-3) caused by ABCB4 gene mutations, and it also enriched the ABCB4 pathogenic variant spectrum. Whole-exome sequencing technology provides a reliable diagnostic tool for etiological analysis.

Extracellular vesicles (EVs) are small bilayer lipid membrane vesicles that can be released by most cell types and detected in most body fluids. EVs exert key functions for intercellular communication via transferring their bioactive cargos to recipient cells or activating signaling pathways in target cells, and hence participate in the variety of diseases including the occurrence and development of liver diseases. In recent years, the prevalence of nonalcoholic fatty liver disease (NAFLD) has increased. Currently there is no reliable method except invasive liver biopsy for the diagnosis of liver inflammation or fibrosis staging. Therefore, the search for the corresponding markers of noninvasive circulation continues to be active, and extracellular vesicles are one of the most concerned. To this end, we reviewed current knowledge about the physical characteristics, biological components, and isolation methods of extracellular vesicles, and introduced the concept of using circulating cell-derived vesicles as a new diagnostic marker for nonalcoholic fatty liver disease.

Objective To compare the efficacy and safety of 12 weeks-entecavir (ETV) treatment in HBeAg-negative chronic hepatitis B patients with acute-on-chronic liver failure (HBV-ACLF) in comparison to lamivudine (LAM).Methods Ninety eight HBeAg-negative patients with HBV-ACLF who were nucleos(t) ide analogs treatment naive as well as with detectable serum HBV DNA were randomly divided into ETV group and LAM group,and each have 49 patients.Additional to the comprehensively basic internal medicine treatment,antiviral therapy with ETV (0.5 mg,qd) or LAM (0.1,qd) was performed,respectively.The differences of mortality rates,clinical improvement rates,complete virological response (CVR) rates and,adverse events between ETV group and LAM group were compared after 12 weeks of treatment.Results The baseline characteristics of patients in ETV group were comparable to those in LAM group.At week 12,a lower mortality rate of 28.6％ than that of LAM group (48.9％) was observed in ETV group (P ＜ 0.05).In terms of clinical improvement rate,ETV group indicated a higher tendency in total (65.3％ vs 48.3％,P =0.067) and a statistically significant value in subpopulation of model for end-stage liver disease scored no more than 30 than LAM group (75.6％ vs 52.5％,P ＜ 0.05).As expected,ETV group achieved a higher CVR rate than LAM group at week 12 (94.3％ vs 72％,P ＜0.05).Discontinuation of antiviral therapy occurred to none of the patients in both groups.Conclusions Comparing with LAM,ETV can safely inhibit HBV replication more intensively and reduce the 12 weeks mortality rate in HBeAg-negative patients with HBV-ACLF.

OBJECTIVETo develop and evaluate a mouse model of nonalcoholic steatohepatitis (NASH) induced by a high-fat and high-fructose (HFHFr) diet.

METHODSSix-week-old C3H mice were randomly divided into groups for HFHFr diet experimental modeling, high fat-only (HF) diet controls, high fructose-only (HFr) diet controls, and standard chow (SC) diet controls. The standard HFHFr diet was modified so that it consisted of 76.5% standard chow, 12% lard, 1% cholesterol, 5% egg yolk powder, 5% whole milk powder, and 0.5% sodium cholate, along with 20% fructose drinking water. At the end of experimental weeks 4, 8, and 16, measurements were taken for the NASH-related parameters of body mass, serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), lipid profile, and wet liver weight (upon sacrifice). In addition, histological changes in the liver were evaluated by hematoxylin-eosin (HE) and oil red O staining. The significance of differences between groups was assessed by statistical analysis, using the

METHODSof t-test, Wilcoxon rank sum test, x2 test, F test or Fisher's test as appropriate.

RESULTSAs compared to the mice in the SC group at the corresponding time points, the mice in the HFHFr and HF groups showed significantly higher body mass and wet liver weight, as well as more extensive and robust lipid disposition in hepatic tissues as evidenced by oil red O staining. However, HE staining indicated that the HFHFr and HF groups had different degrees of macrosteatosis accompanied with intralobular inflammatory foci, with the former showing more remarkable NASH-related histological changes. Analysis at the end of week 16 showed that about 80% of the mice in the HFHFr group had developed NASH [nonalcoholic fatty liver disease (NAFLD) activity score (NAS): less than 5]. The levels of low-and high-density lipoprotein (LDL and HDL) cholesterol, as well as the levels of ALT and AST, were increased from the end of week 4 to the end of week 8 for the HFHFr and HF groups. At the end of week 16, the two groups differed in the extent of increase in total cholesterol and LDL and HDL cholesterol, with only the HFHFr group showing statistically significant changes. Specifically, at the end of week 16, the HFHFr group showed ALT levels of 108.5 +/- 93.34 U/L (F=5.099, P =0.005 vs. HF group: 44.30 +/- 35.71 U/L, HFr group: 46.70 +/- 17.95 U/L, SC group: 24.70 +/- 6.57 U/L), AST levels of 316.30 +/- 208.98 U/L (F=6.654, P=0.001 vs. HF: 132.12 +/- 75.43 U/L, HFr: 143.30 +/- 38.53 U/L, SC: 122.60 +/- 12.76 U/L), total cholesterol levels of 5.18 +/- 0.58 mmol/L (F=72: 470, P =0.000 vs. HF: 3.94 +/- 0.75 mmol/L, HFr: 2.30 +/- 0.50 mmol/L, SC: 2.02 +/- 0.24 mmol/L), HDL cholesterol levels of 3.05 +/- 0.49 mmol/L (F=25.413, P =0.000 vs. HF: 2.65 +/- 0.54 mmol/L HFr: 1.77 +/- 0.47 mmol/L, SC: 1.58 +/- 0.16 mmol/L), LDL cholesterol levels of 1.11 +/- 0.23 mmol/L (F =83.297, P =0.000 vs. HF: 0.72 +/- 0.17 mmol/L, HFr: 0.27 +/- 0.04 mmol/L, SC: 0.20 +/- 0.05 mmol/ L).

CONCLUSIONThe present study suggests that a mouse model of NASH can be successfully induced by a 16-week modified HFHFr diet.

Objective: To investigate the role of neutrophil elastase inhibitor, sivelestat, in preventing and treating nonalcoholic steatohepatitis (NASH) and its underling mechanisms. Methods: A total of forty 4-week-old male C57BL/6J ApoE-/-mice were equally divided into the following four groups: standard chow (SC)+isotonic saline; SC+sivelestat; high-fat, high-cholesterol (HFHC) diet+isotonic saline; and HFHC+sivelestat. These mice were treated with above methods for 12 weeks. Blood and liver tissue samples were collected to measure biochemical parameters, hepatic steatosis and non-alcoholic fatty liver disease (NAFLD) activity score (inflammation) were evaluated by oil red O staining and HE staining, respectively. The mRNA and protein expression levels of hepatic inflammatory cytokines, CD68, and F4/80 were determined by quantitative RT-PCR and immunohistochemistry, respectively. Comparison of means between the four groups was made by one-way analysis of variance, and comparison between any two groups was made by the LSD or SNK method (for data with homogeneity of variance) or the Tamhane or Dunnett method (for data with heterogeneity of variance). Results: Mice fed with an HFHC diet for 12 weeks developed typical pathological features of NASH compared with those fed with SC. Compared with mice fed with HFHC diet without sivelestat, those treated with HFHC and sivelestat exhibited the following features: (1) significantly reduced fast blood glucose, blood cholesterol, and hepatic biochemical parameters, as well as increased insulin sensitivity; (2) significantly reduced NAFLD activity score (5.71±1.11 vs 3.16±1.16, P < 0.05); (3) reduced monocyte chemoattractant protein-1 and tumor necrosis factor -α; (4) significantly reduced mRNA levels of CD68 and F4/80; and (5) reduced expression of CD68 in the liver. Conclusion Sivelestat alleviates the hepatic steatosis and inflammation of NASH in mice by inhibiting the activation of Kupffer cells.