Nonalcoholic fatty liver disease (NAFLD) has become one of the research hotspots in the field of liver disease. However, so far, no drugs have been approved by the U.S. Food and Drug Administration for the treatment of nonalcoholic steatohepatitis, which brings opportunities and challenges to the clinical trials on the treatment of NAFLD. Liver histology is currently considered a reliable surrogate endpoint for tracking the progression of NAFLD, but its invasiveness has limited the development of drugs for the treatment of NAFLD. In recent years, some noninvasive measurement methods have gradually been used as secondary or exploratory endpoints in existing clinical trials.

Objective To explore the effect of NE/α1-AT on the pathogenesis of NASH and the adjustment function of berberine to the imbalance of NE/α1-A and the therapeutic action of berberine to NAFLD.Methods 8-week-old male C57BL/6JApoE/ mice (26),according to the weight assigned to the high-fat-high-cholesterol group (18) and standard chow group (8).HFHC group fed with high fat high cholesterol diet and normal water;the SC group fed with normal diet and drank normal water.After 8 weeks,the HFHC group divided into HFHC group and BBR group according to the weight,Berberine group with 200mg/kg/d berberine,the other two groups fed with the same volume of distilled water.12 weeks all mice were sacrificed.We detected body weight,live weight,live function and lipid metabolism.HE and oil red O staining were used to evaluate pathological changes.ELISA method was used to detect NE and α1-AT expression in mice live tissue;RT-PCR method was used to detect protein α1-AT mRNA expression in live tissue; Immunohistochemistry and Western Blotting were applied to determine expression of NE.Results Compared with the SC group at the end of 12 weeks,ALT,AST were significantly increased in HFHC group,TG was significantly higher than the SC group.Body weight and live weight of ApoE-/-mice fed with HFHC diet significantly higher than the SC group.HE staining showed obvious steatosis and inflammatory foci in HFHC group.The NAS score at the end of model establishment was 5-8 points,which reached the diagnostic criterion for NASH.The concentration and activity of NE in the SC group were significantly increased,and the level of α1-AT was decreased,which lead to ratio NE/α1-AT was significantly higher than the SC group.BBR in mice treated with HE staining showed inflammation ballooning lower than HFHC group.At week 12,ALT,AST of BBR group increased compared with the SC group,and significantly lower compared with the HFHC group.NE/α1-AT was significantly higher than the HFHC group.Conclusions 12 weeks of high fat and high cholesterol feeding,ApoE-/-mice can successfully establish the NASH mouse model;a marked change and contact exist between NE and α1-AT in the process of development of NASH; Berberine can reduce liver lipid deposition and the degree of inflammation in NASH mice,and can improve the NE/ alpha 1-AT imbalance.

Objective To investigate the value of hydrogen proton MR spectroscopy (1H-MRS) and Dixon sequence for the quantitative diagnosis and classification of steatosis in patients with non-alcoholic fatty liver disease (NAFLD). Methods Sixty seven patients with NAFLD confirmed by liver biopsy were prospectively collected from October 2015 to May 2017 in Hangzhou Normal University Hospital. All patients underwent 1H-MRS and Dixon sequence scan within 7 days after liver biopsy, 1H-MRS-based hydrogen proton density fraction (MRS-PDFF) and Dixon-based hydrogen proton density fraction (MRI-PDFF) were obtained. Fat grading based on the fat percentage obtained from liver biopsy. Pearson correlation analysis was performed to analyze the correlation among pathological steatosis and MRS-PDFF, MRI-PDFF. One-way ANOVA analysis was performed to compare the difference of PDFF between patients with different degrees of severity of fatty liver. And the ROC curve analysis was performed to generate the thresholds of MRS-PDFF and MRI-PDFF for determining the presence of fatty liver. Results The steatosis grade of pathological biopsy showed grade S1 in 36 cases, grade S2 in 16 cases, grade S3 in 15 cases, the MRS-PDFF values of S1, S2 and S3 patients were (8.25 ± 4.32)％, (15.67 ± 4.54)％, (23.46 ± 5.82)％and the MRI-PDFF values were (6.31 ± 2.94)％, (15.42 ± 5.07)％, (24.47 ± 6.31)％. Statistically significant differences were observed among them (P<0.01). Both MRS-PDFF and MRI-PDFF were positively correlated with histological fat percentage (r values were 0.840 and 0.892,all P<0.01), there was also a correlation between MRS-PDFF and MRI-PDFF (r=0.930, P<0.01). Area under ROC curve of MRS-PDFF and MRI-PDFF for differential diagnosis of grade S1 steatosis were 0.955 and 0.976, and area under ROC curve for differential diagnosis of grade S3 steatosis were 0.972 and 0.978. Conclusion 1H-MRS and Dixon sequces have high value in liver fat content detection and classification of patients with NAFLD, and both have similar diagnostic efficacy.

Objective:To investigate the accuracy of magnetic resonance imaging (MRI) for quantitative determination of liver fat and iron content through a rat model of non-alcoholic fatty liver disease (NAFLD) induced by methionine-choline deficient (MCD) diet.Methods:Sixty SD rats were randomly divided into experimental (MCD-diet group, n = 30) and normal control group (normal diet, n = 30). Rats were subjected to special MRI examinations at the ends of 2, 4, and 8 weeks. Proton density fat fraction (PDFF) and R2* value were obtained, and then the rats were sacrificed. The liver tissues were stained with HE, Prussian blue, etc. Liver tissue non-heme iron (NHI) homogenate was determined by flame atomic absorption spectrometry. According to different data, one-way analysis of variance, t-test or χ2 test was used for statistical analysis. Results:PDFF and R2 * values in the MCD diet group at 2, 4 and 8 weeks were 23.37% ± 9.20%, 28.07% ± 6.84%, 25.40% ± 7.04% ( P < 0.01) and 90.58 ± 15.92, 104.12 ± 13.47, 106.35 ± 15.76 ( P < 0.05), respectively, which were significantly higher than the normal control group PDFF (2.39% ± 0.50%, 2.45% ± 0.45%, 3.26% ± 0.80%) and R2* (48.93 ± 7.90, 54.71 ± 5.91, 64.25 ± 15.76). Additionally, with the disease progression, R2 * had gradually increased, which was consistent with the NHI trend in liver tissue homogenates of each group. Conclusion:MRI, as a non-invasive quantitative method, can accurately assess liver fat and iron content in fatty liver disease, and with the degree of severity of fat changes, iron deposits tend to increase.

Objective:To analyze the correlation between serum ferritin and steatosis in non-alcoholic fatty liver disease.Methods:Data of 167 cases who underwent liver biopsy in the Affiliated Hospital of Hangzhou Normal University were collected. Hydrogen proton magnetic resonance spectroscopy were performed within one week. The pathological results of liver biopsy were used as the gold standard to analyze the case data, serological indicators, magnetic resonance spectroscopy-proton density fat fraction.Results:Pathological monitoring result showed that the serum ferritin in patients without steatosis, and with mild, moderate and severe steatosis were (206.20 ± 189.83), (286.65 ± 200.80), (326.55 ± 214.71), (391.50 ± 184.93) ng/ml, respectively, P < 0.005. Serum ferritin was correlated to body mass index, PDFF, alanine aminotransferase, gamma glutamyltransferase, low-density lipoprotein, high-density lipoprotein. The area under ??the receiver operating characteristic curve with ferritin for the diagnosis of non-alcoholic fatty liver disease was 0.716, and the optimal diagnostic threshold was 214.56 ng/ml. The sensitivity and specificity were 80.1%, and 68.8%, respectively. There was no statistically significant difference between the intralobular inflammation, fibrosis, and ferritin. Prussian blue iron staining had no apparent deposition of iron particles. Conclusion:Ferritin has significant positive correlation with the results of pathological and magnetic resonance imaging for liver steatosis. Therefore, it can be used as a non-invasive diagnostic method for liver steatosis evaluation.

Objective:To explore the relationship between liver iron deposition and steatosis in patients with non-alcoholic fatty liver disease (NAFLD) through MRI.Methods:163 cases of liver biopsy underwent MRI examination. R2* was used to measure liver iron content. Dixon-based proton density fat fraction (PDFF) was used to measure liver fat content. One-way ANOVA, r-correlation, ROC curve, and others were used to assess the relationship between clinical case data, serological indices, and imaging results in accordance with the pathological results of the liver biopsy.Results:R2* gradually increased as the pathological steatosis grade rose. The R2* that corresponded to no steatosis (< 5%), mild steatosis (14.95%±8.55%), moderate steatosis (46.30%±9.32%), and severe steatosis (73.86%±6.35%) were 27.56±4.40, 31.06±5.95, 38.06±4.80, and 48.10±5.55 ( P < 0.001), respectively. There was a positive correlation between R2* and liver steatosis content ( r= 0.769, P < 0.05). The area under the ROC curve and cut-off value were 0.88 and 31.77, respectively, and there was no distinct relationship with liver inflammation or fibrosis. Conclusion:R2* can quantitatively and non-invasively evaluate liver iron deposition in patients with NAFLD. A distinct relationship exists between liver steatosis and iron deposition, and iron deposition tends to increase as the steatosis aggravates.

Objective: We aimed, in our prospective study, to assess the predictive value of serum non-invasive and biochemical markers for clinical diagnosis of significant fibrosis (including early stages). Methods: We measured sH2a levels in serum, comparing with routine liver function markers. We compared blindly pretreatment serum samples from a cohort of hepatitis B patients without non-alcoholic fatty liver disease(NAFLD), which had histological grades of liver fibrosis, with NAFLD individuals and CHB with NAFLD patients. Statistical analysis was by Student′s t test, and receiver-operating characteristic (ROC) curves were drawn. Results: ROC curves showed that serum sH2a had greater diagnostic performance than routine liver function markers compared with histological grades of liver fibrosis(S0, S1-2, S3-4). ROC curves showed that using a sH2a cut-off point of 0.79 was with highest sensitivity as 63% and highest specificity as 80%. And sensitivity as 96.7% and specificity as 75.5% when using a sH2a cut-off point of 0.77. Conclusions sH2a has the potential to be a uniquely sensitive and specific novel marker for liver fibrosis and function.

Objective: To investigate the value of 1H-magnetic resonance spectroscopy (¹H-MRS) in determining the content of liver triglyceride in patients with fatty liver disease (FLD), as well as its influencing factors. Methods: A total of 124 patients with nonalcoholic fatty liver disease (NAFLD), chronic hepatitis B (CHB), or hepatitis B complicated by FLD who underwent liver biopsy in the Affiliated Hospital of Hangzhou Normal University were enrolled, and the clinical data, serological markers, FibroScan results, and ¹H-MRS results were collected. A correlation analysis was performed with the results of liver biopsy as the gold standard, and the influence of factors including hepatitic B virus (HBV) infection and obesity on accuracy was analyzed. A one-way analysis of variance was used for comparison of means between the three groups, and the LSD or SNK test (for homogeneity of variance) or the Tamhane’s or Dunnett’s test (heterogeneity of variance) was used for comparison between any two groups. The t-test was used for comparison of continuous data between groups, and the chi-square test was used for comparison of categorical data. The MRS-PDFF receiver operating characteristic (ROC) curve was plotted, the area under the ROC curve (AUC) was calculated, the optimal cut-off points for the diagnosis of NAFLD were estimated, and sensitivity and specificity were calculated. Results: The NAFLD group (42 patients) and the CHB + NAFLD group (40 patients) had a significantly higher proton density fat fraction (PDFF, the content of triglyceride in the liver) than the CHB group (42 patients) (16.84±9.76/9.39 ± 5.50 vs 3.45 ± 1.63, P < 0.001). The results were significantly correlated with the degree of steatosis confirmed by liver biopsy (P < 0.001), but it was not significantly correlated with inflammation or fibrosis grade. The correlation analysis showed that the MRS-PDFF value measured by 1H-MRS was significantly correlated with body mass index (BMI), blood lipids, alkaline phosphatase, and blood glucose, while it was not significantly correlated with age, sex, or the presence or absence of hepatitis B. The ROC curve analysis showed that the AUCs of PDFF measured by 1H-MRS were 0.93, 0.974, and 0.976, respectively, for the diagnosis of steatosis S1(≥5%), S2(≥34%), and S3(≥66%), and the corresponding optimal thresholds were 5.14%, 11.16%, and 16.7%, respectively. Conclusion 1H-MRS has a high diagnostic value in quantitative evaluation of the degree of liver steatosis in patients with FLD and is not affected by the factors such as HBV infection, age, and sex, while it is correlated with BMI and lipid metabolism.