Mild Fibrosis Group Research Articles

Metabolic Changes in Different Stages of Liver Fibrosis: In vivo Hyperpolarized 13C MR Spectroscopy and Metabolic Imaging.

The objective was to assess metabolic changes in different stages of liver fibrosis using hyperpolarized C-13 magnetic resonance spectroscopy (MRS) and metabolic imaging. Mild and severe liver fibrosis were induced in C3H/HeN mice (n = 14) by injecting thioacetamide (TAA). Other C3H/HeN mice (n = 7) were injected with phosphate buffer saline (PBS) (7.4 pH) as normal controls. Hyperpolarized C-13 MRS was performed on the livers of the mice, which was accompanied by intravoxel incoherent motion (IVIM) diffusion-weighted imaging with 12 b values. The differential metabolite ratios, apparent diffusion coefficient values, and IVIM parameters among the three groups were analyzed by a one-way analysis of variance test. The ratios of [1-13C]lactate/pyruvate, [1-13C]lactate/total carbon (tC), [1-13C]alanine/pyruvate, and [1-13C] alanine/tC were significantly higher in both the mild and severe fibrosis groups than in the normal control group (p < 0.05). While the [1-13C]lactate/pyruvate and [1-13C]lactate/tC ratios were not significantly different between mild and severe fibrosis groups, the ratios of [1-13C]alanine/pyruvate and [1-13C]alanine/tC were significantly higher in the severe fibrosis group than in the mild fibrosis group (p < 0.05). In addition, D* showed a significantly lower value in the severe fibrosis group than in the normal or mild fibrosis groups and negatively correlated with the levels of [1-13C] lactate and [1-13C]alanine. Our findings suggest that it might be possible to differentiate mild from severe liver fibrosis using the cellular metabolic changes with hyperpolarized C-13 MRS and metabolic imaging.

Gadolinium Ethoxybenzyl Diethylene-triamine-pentaacetic-acid-enhanced Magnetic Resonance Imaging Perfusion and Multiple Hepatobiliary-phase Imaging for Quantitative Assessment of Liver Fibrosis in Rat Models

Objective To investigate the value of gadolinium ethoxybenzyl diethylene-triamine-pentaacetic-acid (Gd-EOB-DTPA)-enhanced magnetic resonance imaging (MRI) in detecting different stages of liver fibrosis in rats.Methods Rat models of liver fibrosis were induced by carbon tetrachloride intraperitoneal injection for 4 - 12 weeks (n=45). The control group was applied with 0.9% saline (n=15). The MRI protocol contained both dynamic contrast-enhanced sequence (60 continuous scans within 3 minutes,including three pre-contrast measurements) and multiple hepatobiliary-phase acquisitions (every 5 minutes after contrast injection,60 minutes in total). METAVIR score was used to grade liver fibrosis:normal (F0),mild fibrosis (F1 - F2),and advanced fibrosis (F3 - F4). Liver perfusion parameters [transfer constant (K trans),extravascular extracellular volume fraction (Ve),initial area under curve (iAUC),maximum relative enhancement (REmax),and time of maximum RE (Tmax)] as well as hepatobiliary-phase parameters [RE at different time point,the decrease of RE (REchange=RE3 min - RE60 min),and elimination half-life of RE (TRE1/2)] were measured and compared with ANOVA analysis and Spearman rank correlation.Results Thirty-one rats completed MRI exams and were then divided into normal (n=10),mild fibrosis (n=10),and advanced fibrosis (n=11) groups. K trans,Ve and iAUC decreased as liver fibrosis progressed (r=-0.631,P=0.002;r=-0.503,P=0.017;r=-0.446,P=0.037). K trans and Ve showed significant differences among three groups(F=7.011,P=0.005;F=4.656,P=0.023). K trans and Ve were significantly lower in advanced fibrosis group than in normal group (P=0.001,P=0.009). There were statistical significant differences of Tmax,TRE1/2 and REchange among groups(F=6.633,P=0.005;F=5.493,P=0.010;F=5.343,P=0.014). Compared to normal and mild fibrosis groups,advanced fibrosis group had significantly longer Tmax and TRE1/2 (P=0.005,P=0.004;P=0.008,P=0.008)and significantly lower REchange(P=0.007,P=0.012).Conclusion Perfusion and multi-hepatobiliary-phase parameters such as K trans,Ve,Tmax, TREI/2 and REchange obtained from Gd-EOB-DTPA-enhanced MRI,may be valuable for detecting and staging liver fibrosis.

Non-Proximal Renal Tubule-Derived Urinary Exosomal miR-200b as a Biomarker of Renal Fibrosis

Background: Renal fibrosis is a common outcome of nearly all kinds of chronic kidney disease (CKD) and eventually progresses to end-stage renal disease. The identification of an optimal biomarker of renal fibrosis to replace the invasive renal biopsy will have important clinical implications. Methods: We isolated urinary exosomes from 50 participants and examined the exosomal protein content and particle number in 38 CKD patients with different degrees of renal fibrosis and in 12 normal individuals. We examined the levels of exosomal microRNAs (miRNAs), namely, miR-200a, miR-200b, miR-200c, miR-141, miR-429, miR-29a, miR-29b, miR-29c, miR-192, and miR-21, by sorting the exosomes and comparing the levels of proximal tubular, non-proximal tubular, and total exosomal miR-200b. Results: The exosome content was higher in the CKD group, but no differences were evident among the mild, moderate, and severe fibrosis groups. Among the 10 exosomal miRNAs, miR-200b was lower in the CKD group than in the normal group and decreased more significantly with fibrosis progression as well as in IgA nephropathy and diabetic kidney disease. CD13⁺ CD63⁺ exosomes constituted 18.6% of all urinary exosomes. Sorting the proximal tubular exosomes with the CD13 protein marker revealed that miR-200b in the CD13⁺ group was extremely low; however, the result was significantly different in the CD13^– group but not in the CD13⁺ group. The magnitude of the decline was greater in the CD13^– groups than in the non-sorted whole groups between the fibrosis and normal patients. Conclusions: Non-proximal renal tubule-derived urinary exosomal miR-200b is a biomarker of renal fibrosis. Exosomes can be used as a liquid biopsy and may replace the traditional invasive renal biopsy in the diagnosis of renal fibrosis.

Changes of FibroScan, APRI, and FIB-4 in chronic hepatitis B patients with significant liver histological changes receiving 3-year entecavir therapy.

Noninvasive fibrosis tests have been used widely for evaluation of liver fibrosis in patients with chronic hepatitis B (CHB). We aimed to investigate the influence of antiviral treatment on FibroScan, APRI, and FIB-4 in CHB patients with significant liver histological changes (SLHC) defined as inflammatory grade≥A2 and/or fibrosis stage≥F2. A total of 104 CHB patients with SLHC at the baseline were included. FibroScan, APRI, and FIB-4 values were compared before and after 3-year entecavir (ETV) treatment. Liver stiffness measurement values decreased significantly after 3-year ETV treatment in cirrhosis group (from 13.6 to 9.6kPa, p=0.018), significant fibrosis group (from 8.4 to 5.8kPa, p=0.001), and mild fibrosis group (from 5.5 to 4kPa, p<0.001). APRI decreased significantly after 3-year ETV treatment in patients with cirrhosis (from 0.80 to 0.25, p<0.001), patients with significant fibrosis (from 0.54 to 0.24, p<0.001), and those with mild fibrosis (from 0.35 to 0.23, p<0.001). FIB-4 decreased significantly after 3-year ETV treatment in patients with cirrhosis (from 1.27 to 0.81, p=0.007) and significant fibrosis (from 1.12 to 0.78, p<0.001), while did not decrease significantly in patients with mild fibrosis (from 0.90 to 0.80, p=0.389). FibroScan, APRI, and FIB-4 values decreased significantly after 3-year ETV treatment in CHB patients, whichindicates that these noninvasive fibrosis tests might be useful for monitoring regression of liver fibrosis and assessing treatment efficacy during long-term ETV treatment.

Pathogenesis of non-alcoholic fatty liver disease mediated by YAP.

This study aimed to investigate the mechanism of the interaction between Yes-associated protein (YAP) and transforming growth factor-β (TGF-β)/Smad signaling pathways in the development of non-alcoholic fatty liver disease (NAFLD). Serum samples of monkeys with biopsy-proven NAFLD and healthy normal monkeys were used to measure fasting plasma glucose (FPG), low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglyceride (TG) and albumin (ALB) with the BECKMAN CX5 PRO. Hematoxylin-eosin staining (H&E) was used for pathologic analysis, Masson trichrome staining was used to assess for fibrosis staging, and Oil Red O staining was used to detect lipid droplet deposition. According to an NAFLD activity score of <4 points and >4 points, the samples were divided into groups: the steatosis group and fibrosing NASH group. Furthermore, monkeys with a fibrosis stage <2 were assigned to the mild fibrosis group, while monkeys with fibrosis stage ≥2 were assigned to the significant fibrosis group. Moreover, the fibrosis stage was subdivided as follows: stages 1a, 1c and 2-3. Immunohistochemistry and real-time quantitative PCR were used to quantify protein and gene expression, respectively. In the present study, 54 monkeys with NAFLD and 23 normal monkeys were recruited. Serum FPG and TG levels were higher in fibrosing NASH monkeys compared with simple steatosis and normal monkeys, and differences between simple steatosis and normal monkeys were not statistically significant (p>0.05). YAP increased in NAFLD, which mainly localized in the nuclei of hepatocytes, perivascular cells and bile duct cells; the accumulation of YAP correlated with the severity of hepatocyte injury. Compared with normal monkeys, the expression of TGF-β, α-smooth muscle actin (α-SMA), Drosophila mothers against decapentaplegic protein 3 (Smad3) and connective tissue growth factor (CTGF) in the liver of simple steatosis monkeys significantly increased (p<0.01). Compared with simple steatosis monkeys, the expression of TGF-β, α-SMA, Smad3 and CTGF in fibrosing NASH significantly increased (p<0.01). However, the expression of Drosophila mothers against decapentaplegic protein 7 (Smad7) in the liver of fibrosing NASH monkeys significantly decreased (p<0.01). With the severity of liver fibrosis, the expression of TGF-β, α-SMA, Smad3 and CTGF gradually increased, and the difference was statistically significant (p<0.01). However, there was no significant difference in the expression of Smad3 between fibrosis stage 1a and 1c. Compared with normal monkeys, the expression of Smad7 in the liver of monkeys with fibrosis significantly decreased (p<0.01), but was significantly higher at fibrosis stage 1c than at fibrosis stage 1a and 2. The YAP and TGF-β signaling pathways and the interaction between them promote the development and progression of NAFLD.

Differentiation between cystic biliary atresia and choledochal cyst: A retrospective analysis.

Cystic biliary atresia (CBA) can be easily misdiagnosed as choledochal cyst (CC). Some patients have already progressed to severe liver fibrosis and missed the optimal surgical time, when the differential diagnosis is made. We aim to determine the differentiation between CBA and CC, and to validate the value of aspartate aminotransferase-to-platelet ratio index (APRI) in the assessment of liver fibrosis and prediction of post-operative outcome for infants with biliary cystic malformations (BCMs). Clinical data of children (categorised into CBA and CC groups) with BCMs were analysed retrospectively. Biochemical indicators together with B-ultrasound examinations and the degree of liver fibrosis were analysed, and those with statistical difference between the two groups were selected for diagnostic receiver operating characteristic curve analysis. The parameter that showed the highest accuracy with a significant diagnostic performance for differentiating CBA from CC was cyst size. Liver assessment at operation was categorised into mild fibrosis and moderate-to-severe fibrosis. The APRI values were much lower in the mild fibrosis groups than in the moderate-to-severe fibrosis group (0.4 ± 0.2 vs. 1.4 ± 0.8, P < 0.001). A cut-off value of 0.96 (area under the curve 0.92, P < 0.001) showed a sensitivity of 81.3% and a specificity of 100% for moderate-to-severe fibrosis. Lower APRI value was correlated with short-term post-operative bilirubin clearance. There is still certain difficulty in the early identification of CBA and CC clinically. Liver fibrosis could occur as early as infantile period in both CBA and CC. In infants with BCMs, APRI can be used as a non-invasive method to detect liver fibrosis in early stages.

Fn14 hepatic progenitor cells are associated with liver fibrosis in biliary atresia.

The liver in biliary atresia (BA) is characterized by progressing fibrosis which is promoted by unclear reasons. We aimed to understand the factors influencing liver fibrosis. This study hypothesized that HPCs (hepatic progenitor cells) are activated and associated with liver fibrosis in biliary atresia. Liver samples from biliary atresia patients are as BA group, and the normal liver derived from hepatoblastoma infants during operation are control group. The extent of fibrosis in liver samples was blindly evaluated by two experienced pathologists depending on Ishak system. The BA liver samples were divided into mild liver fibrosis group (grade I-IV, BAa) and severe liver fibrosis group (grade V-VI, BAb) to detect Fn14 protein expression. In mRNA level, Fn14 expression was 21.23 ± 8.3 vs. 1.00 ± 0.17, p = 0.023 < 0.05 and CD133 expression was 6.02 ± 2.16 vs. 1.14 ± 0.75, p = 0.008 < 0.01 between BA group and control group. Fn14 cells co-expressed the progenitor marker CD133 in liver, and activated in BA. Fn14 andα-SMA were co-location in fibrous area in liver. Compared to the control group, Fn14, CD133, and α-SMA protein expression were 2.10 ± 0.53 vs. 0.97 ± 0.2, p = 0.001, 2.23 ± 0.57 vs. 1.00 ± 0.03, p = 0.000, 4.96 ± 2.4 vs. 1.00 ± 0.22, p = 0.001. The Fn14 protein expression was 2.60 ± 0.35 vs. 1.86 ± 0.42, p = 0.012, between BAb and BAa group. Fn14 cells, which co-express the progenitor marker CD133 in liver, are HPCs and activated in BA. Fn14 + HPCs are associated with liver fibrosis in BA.

Integrative Cardiopulmonary Ultrasound for Interstitial Lung Disease Assessment: Correlation between Lung Ultrasound Performance and Cardiac Involvement

The aims of this study were to apply integrative cardiopulmonary ultrasound (thoracic ultrasound) to the evaluation of patients with interstitial lung disease (ILD) and to determine the relationship between lung ultrasound signs and echocardiographic parameters such as systolic pulmonary artery pressure (SPAP) and various right ventricular parameters. ILD patients and healthy controls underwent lung ultrasound (LUS) and echocardiographic tests. In addition to traditional echocardiographic parameters, right ventricular free wall longitudinal strain (RVLS_FW) was measured using 2-D speckle-tracking echocardiography. The degree of pulmonary fibrosis or the disease severity of each ILD patient was scored with a semiquantitative scoring method, taking into account multiple LUS signs. Statistical analyses were performed to compare various parameters between ILD patients with and those without pulmonary hypertension. Correlations between the different parameters were then evaluated, and the LUS scores were used to predict pulmonary hypertension using a receiver operating characteristic curve analysis. Among the 90 patients who qualified for entry into this study, 30% (n = 27) had pulmonary hypertension. The patients with pulmonary hypertension had larger right ventricular dimensions, lower RVLS_FW and higher LUS scores. An effective correlation was found between ILD LUS scores and echocardiographic parameters, especially SPAP (r = 0.735, p < 0.001). The groups were comparable with respect to most echocardiographic parameters, with mild, moderate and severe pulmonary fibrosis, whereas SPAP was significantly higher in the moderate and severe groups. RVLS_FW was obviously reduced in the group with severe pulmonary fibrosis. Although RVLS_FW in the mild pulmonary fibrosis group was in the normal range, it was slightly reduced compared with that of the controls. A cutoff of more than 16 LUS points had 85.2% sensitivity and 80.3% specificity in predicting elevated SPAP (>36 mm Hg). Thoracic ultrasound is useful in the assessment of ILD. As ILD progresses, the structure or function of the right ventricle gradually deteriorates. LUS not only detects lung conditions in ILD, but also indirectly reflects SPAP and right ventricular function. Integrated use of LUS and echocardiography will provide an invaluable point-of-care imaging modality to facilitate the diagnosis, management and treatment of patients with ILD.

Serum Osteopontin and Cytokeratin-18 in Chronic Hepatitis C Patients

Chronic hepatitis C is a global health problem; most patients are at risk for developing fibrosis and cirrhosis. Histological examination of liver biopsies is currently the gold standard for the detection of early liver damage, but there is a strong need for better noninvasive methods. The aim of this study was to evaluate the association between serum osteopontin (OPN) level, serum cytokeratin 18 M30 (CK-18 M30) neoepitope level and the histological severity of hepatic fibrosis in hepatitis C virus (HCV) induced patients. Subjects and methods: This study included 89 subjects, 70 with chronic hepatitis C virus infection, they were classified into 2 groups according to METAVIR fibrosis stage as follows: group I (stage 2 or less was considered as mild liver fibrosis) included 50 patient, group II (stage 3 or more was considered as extensive fibrosis) included 20 patients and 19 healthy matched age and gender as a control group. All subjects were submitted to the following: Through history taking, complete clinical examination, and serum concentrations of osteopontin and cytokeratin 18 M30 neoepitope were measured by enzyme linked immunosorbent assay (ELISA). The results revealed that there were high significant differences of OPN and CK-18 M30 between patients and control (P<0.001). There was a high significant difference of OPN (P<0.001) and a significant difference of CK-18 M30 (P=0.02) when compared between mild fibrosis and extensive fibrosis groups. There was a high significant correlation of serum OPN concentrations with severity of liver fibrosis degree (r=0.75, P<0.001), while the serum CK-18 M30 concentrations showed a significant correlation (r=0.33, P=0.005). In ROC curve serum OPN at the cut-off point of 3.1 ng/ml could discriminate mild from extensive fibrosis with sensitivity of 95%, serum CK-18 M30 at the cut-off point of 293 ng/ml could discriminate mild from extensive fibrosis with sensitivity of 70%. Finally from obtained study, results showed that serum OPN levels was better than CK-18 M30 in identification the degree of hepatic fibrosis and could be used as a biomarker to assess the stage of fibrosis in HCV patients which would help to reduce the number of liver biopsies.

A novel score for diagnosis of liver fibrosis based on Th17 activity and sera fibrosis biomarkers

Background: Fibrosis is a major cause of morbidity and mortality worldwide. Biopsy is an invasive procedure. For the evaluation of liver fibrosis, an alternative, noninvasive, new, easy, and available method is determined to assess hepatic fibrosis in Egyptian patients. Methodology: We enrolled 143 patients with fibrosis and they were divided into two groups, severe fibrosis and mild fibrosis group. Th17 cells were stimulated in vitro with S. mansoni soluble egg antigen (SEA), then interleukin (IL)-17, IL-22, and interferon (IFN)-γ were assessed in the culture supernatant. Serum levels of alanine aminotransferase, aspartate aminotransferase, YKL-40, and hyaluronic acid (HA) were also assessed using enzyme-linked immunosorbent assay. Analysis of individual data was conducted to characterize the diagnostic accuracy of the three highly significant fibrosis biomarkers (HA, IL-22, and IFN-γ). Results: A highly significant difference in IL-22, IFN-γ, and HA levels between mild and severe groups was observed. Linear combination of the three biomarkers was selected by the multivariate discriminate analysis as the best combination for construction of the fibrosis discrimination equation. Application of the equation on all patients correctly classified 28.7% with severe fibrosis and 71.3% with mild fibrosis at a discriminant cut-off score (0.297) with 87.18% sensitivity and 85.57% specificity. Conclusion: A simple fibrosis score = [0.594 (numerical constant) + 0.007 × HA (ng/ml) + 0.006 × IL-22 (ng/ml) − 0.129 × IFN-γ (pg/ml)] may be useful for discrimination of severe from mild fibrosis patients without the need of liver biopsy.

Assessment of fibrotic tissue and microvascular architecture by in-line phase-contrast imaging in a mouse model of liver fibrosis.

To explore the value of in-line phase-contrast imaging with computed tomography (ILPCI-CT) by synchrotron radiation (SR) for liver fibrosis. Liver fibrosis models were set up in 13 BALB/c mice by peritoneal injections of thioacetamide and evaluated by ILPCI-CT. Histological staging was used to categorize liver fibrosis into normal, mild fibrosis and advanced fibrosis groups. Microvessel density (MVD), the ratio of total vessel length to volume (L/V), the ratio of total number of branching points to liver volume (P/V) and the distribution of vessel diameter were assessed. The CT images showed slightly high-density shadows around the portal tracts in the fibrosis group. Three-dimensional reconstruction can detect vascular and nodular changes on the surface of fibrotic livers. The MVDs between the three groups were significantly different (P = 0.024). L/V was significantly different between the three groups (P = 0.014). There was a positive correlation between MVD and P/V. Fibrous material can be detected by ILPCI-CT even in the early stage of fibrosis. MVD, L/V, P/V and the distribution of vessel diameter were consistent with fibrosis-related angiogenesis progress. Three-dimensional reconstruction is a promising method to visualize morphological changes of the fibrotic liver. • ILPCI-CT can detect fibrous material even in the early stage of liver fibrosis. • MVD, L/V, P/V, and the distribution of vascular diameter reflect pathological angiogenesis. • 3D reconstruction could be a promising approach for detecting liver fibrosis.

Serum β-trophin level as a new marker for noninvasive assessment of nonalcoholic fatty liver disease and liver fibrosis.

Nonalcoholic fatty liver disease (NAFLD) is a common chronic liver disease and evaluation of fibrosis is important. We aimed to investigate the utility of serum β-trophin in NAFLD and its ability to predict liver fibrosis. Serum samples of consecutive patients with biopsy-proven NAFLD and age-matched and sex-matched healthy controls were used to measure β-trophin using ELISA. Correlations between histopathological features of NAFLD and β-trophin were analyzed. Whereas patients with fibrosis scores less than 2 were grouped in the mild fibrosis group, patients with scores of 2 or more were grouped in the significant fibrosis group. Univariate/multivariate logistic regression analyses were carried out to evaluate the independent predicting factors of liver fibrosis. Receiver operating characteristics (ROCs) were assessed to determine the best cut-off values for NAFLD and fibrosis. Sixty-nine patients with NAFLD and 69 healthy controls were enrolled in the study. Serum β-trophin levels were lower in NAFLD patients compared with the controls (2.34±0.06 vs. 1.94±0.09 ng/ml, respectively, P<0.001). In NAFLD, serum β-trophin was related to liver fibrosis and inflammation. The mild fibrosis group had higher serum β-trophin levels than the significant fibrosis group (2.11±0.12 vs. 1.72±0.11, respectively, P<0.001). In multivariate analysis, β-trophin remained an independent predictor of significant fibrosis (odds ratio, 0.237; 95% confidence interval, 0.059-0.949; P<0.001). ROC analysis showed that serum β-trophin was statistically significant in the identification of significant fibrosis (area under receiver operating characteristic, 0.844; 95% confidence interval, 0.718-0.970; P<0.001). The best cut-off value was 1.786, with the best sensitivity (71.43%) and specificity (95.65%). Serum β-trophin may be a potential noninvasive marker for the identification of NAFLD and significant liver fibrosis.

Evaluation of the Role of Liver and Splenic Transient Elastography in Chronic Hepatitis C Related Fibrosis

Background: Liver fibrosis is common consequence of chronic HCV infection. Noninvasive assessment is still evolving. Aim: Evaluate role of liver stiffness measurement (LSM), spleen stiffness measurement (SSM) and their combination (CLSM) using FibroScanTM in assessment of liver fibrosis in CHC patients. Methods: 420 CHC patients and 40 healthy controls included.Liver, renal function tests, CBC and INR done. Liver biopsy done for all of them except if contraindicated. Fibrosis was graded by Metavir score. Abdominal ultrasonography was done before the FibroScanTM and the liver biopsy. LSM, SSM, and CLSM had doneusing FibroScanTM in the supine position after 6-8 hours fasting.The patient were classified into mild fibrosis (F1-F2, n=248) and significant fibrosis (F3-F4, n=172) group. Results: There were significant difference (p=0.001) between patients with mild fibrosis (F1-F2) versus patients with significant fibrosis (F3-F4) regarding; the age (35.06 ± 8.63 vs 43.71 ± 7.97 years), serum bilirubin (0.73 ± 0.25 vs 1.26 ± 0.73 mg/dL), serum albumin (4.42 ± 0.32 vs 3.84 ± 0.51 g/dL), platelets (206.81 ±50.55 vs 140.50 ± 53.77×103/μL), platelets spleen ratio (1762.20± 521.26 vs. 1014.64 ± 470.27). Furthermore a significant difference (p=0.001) detected with LSM (6.57 ± 2.62 vs. 23.04± 12.15 kPa), SSM (25.56 ± 5.36 vs 46.19 ± 16.29 kPa), and CLSS (32.13 ± 7.15 vs. 69.23 ± 25.43 kPa) respectively. Thecutoff values of significant fibrosis were 9.15 kPa by LSM (94.8% Sensitivity, 88.3% Specificity, 84.9% PPV, and 96.1.8% NPV), 27.5 kPa by SSM (94.8% Sensitivity, 70.2% specificity,68.8% PPV, and 95.1% NPV) and 40.85 kPa by CLSM (92.4% Sensitivity, 91.1% specificity, 87.8% PPV, and 94.6% NPV). Conclusion: The measurement of liver, spleen stiffness by FibroScanTM or their combination is convenient for liver fibrosis assessment.

Impact of hepatic immunoreactivity of angiotensin-converting enzyme 2 on liver fibrosis due to non-alcoholic steatohepatitis

We aimed to evaluate the hepatic immunoreactivity of angiotensin-converting enzyme 2 (ACE2) in non-alcoholic steatohepatitis (NASH) patients, elucidate its association with the clinicopathological characteristics and also determine its role in fibrosis progression. The consecutive biopsy proven NASH patients were subdivided into two groups according to their fibrosis score. Fibrotic stages<3 in mild fibrosis group and fibrotic stages ≥ 3 in advanced fibrosis depending on the presence of bridging fibrosis. Liver biopsy specimens were immunohistochemically stained for ACE2 immunoreactivity. Demographics and clinical properties were compared between the groups. Univariate and multivariate analysis were also performed to evaluate the independent predicting factors for the presence of advanced liver fibrosis caused by NASH. One hundred and eight patients were enrolled in the study. Out of this, ninety-four patients representing 87% were classified as mild fibrosis group, whilst fourteen representing 13% were in advanced fibrosis group. We compared high hepatic immunoreactivity of ACE2 between mild and advanced fibrosis groups and found a statistically significant difference 65.9% vs 28.5%, respectively and P=0.008. Hepatic ACE2 immunoreactivity was inversely correlated with the fibrosis score (r: -0.337; P<0.001). The significant variables in the univariate analysis were then evaluated in multivariate logistic regression analysis and hepatic ACE2 immunoreactivity was an independent predicting factor of liver fibrosis [odds ratio (OR): 0.194; 95% confidence interval (CI): 0.082-0.897, P=0.036]. Hepatic immunoreactivity of ACE2 was inversely correlated with the liver fibrosis among biopsy proven NASH patients and it was also an independent predicting factor of advanced fibrosis.

The role of interleukin-6 and interleukin-8 gene polymorphisms in non-alcoholic steatohepatitis.

Background:Genetic polymorphisms may play role in the pathophysiology of nonalcoholic steatohepatitis (NASH).Objectives:We purposed to assess the role of interleukin 6 (IL 6) and interleukin 8 (IL 8) gene polymorphisms in the pathogenesis of NASH.Patients and Methods:Consecutive patients with biopsy proven NASH and age- and gender-matched healthy individuals with normal liver function tests and normal ultrasonography were enrolled in the study. Histopathological findings were recorded according to nonalcoholic fatty liver disease activity score (NAS). Patients were classified according to fibrosis scores as fibrosis score < 2 (mild fibrosis group) and fibrosis score ≥ 2 (significant fibrosis group). Blood samples were collected and genomic DNA isolation kit was used to evaluate genetic polymorphisms.Results:Of thirty-eight patients, 27 (71%) were in mild fibrosis group and 11 (29%) in significant fibrosis group. Thirty-eight age- and gender-matched healthy controls were enrolled in the study. The frequencies of genotypes G/C and G/G of IL 6 among the NASH group and healthy controls were 39.5% and 60.5% vs. 53.6% and 46.4%, respectively (P = 0.32). The frequencies of the genotypes of IL 8 among the NASH group were 47.2%, 44.6%, and 8.2% for T/T, A/T, and A/A, and in healthy controls were 50%, 28.6% and 21.4%, respectively, (P = 0.568). The differences between IL 8 gene T/A and T/T genotypes were not significant statistically (P > 0.05). However, the frequency of A/A genotype in significant fibrosis group was higher than the mild fibrosis group (P = 0.0016). The differences of -251 A/T polymorphism in the IL 8 and -174 C/G polymorphism in the IL 6 were not statistically significant between fibrosis groups (P > 0.05).Conclusions:IL6 and IL8 gene polymorphisms have no role in NASH pathogenesis and liver fibrosis process, but presence of the A/A genotype in the IL8 gene is associated with disease progression.

Estrogen receptor alpha expression and liver fibrosis in chronic hepatitis C virus genotype 1b: a clinicopathological study.

Background:Hepatic damage due to chronic hepatitis C virus (HCV) genotype 1b infection varies widely.Objectives:We aimed to investigate whether estrogen receptor alpha (ERα) plays a role in liver fibrosis in patients infected with HCV genotype 1b.Patients and Methods:All the consecutive patients who received the same standard treatment protocol for HCV genotype 1b were subdivided into two subgroups according to their fibrosis scores as fibrotic stages < 2 in mild fibrosis group and fibrotic stages ≥ 2 in advanced fibrosis group, depending on the presence of septal fibrosis. ERα was stained in liver biopsy specimens. Demographics and clinical properties were compared between the groups. Multivariate logistic regression analysis was performed to predict advanced fibrosis.Results:There were 66 patients in the mild fibrosis group and 24 in the advanced fibrosis group. Among the mild and advanced fibrosis groups, 65.1% and 50%were female, respectively (P = 0.19). There was an inverse correlation between ERα and fibrotic stage (r: -0.413; P < 0.001). Age, platelet counts, neutrophil counts, Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Gamma glutamyl transferase (GGT) and ERα were statistically significant in the univariate analysis. In multivariate logistic regression analyses, ERα expression continued to be an independent predicting factor of liver fibrosis in patients infected with chronic HCV genotype 1b (OR: 0.10; 95% CI: 0.018-0.586; P < 0.001).Conclusions:ERα expression in liver was inversely correlated with liver fibrosis among patients infected with chronic HCV genotype 1b.

Can Serum Pin1 Level Be Regarded as an Indicative Marker of Nonalcoholic Steatohepatitis and Fibrotic Stages?

Background: We aimed to investigate serum Pin1 as an indicator of the presence of nonalcoholic steatohepatitis (NASH) and its association with the histopathological liver fibrosis stages. Methods: Serum samples were collected from consecutive biopsy-proven NASH patients and healthy controls, and then serum levels of Pin1 were measured. The correlations between clinical and histopathological features of NASH and Pin1 were evaluated. Patients who had fibrotic stages <2 were termed mild fibrosis group and those who had ≥2 as advanced fibrosis group. We performed univariate and multivariate logistic regression analyses to evaluate the independent predicting factors for the presence of liver fibrosis caused by NASH. Results: Fifty-six consecutive NASH patients and 56 age- and sex-matched healthy controls were enrolled in the study. Serum Pin1 levels were significantly higher in NASH patients (39.24 ± 30.94) than in controls (27.7 ± 9.56, p < 0.001). In NASH patients, serum Pin1 levels were correlated with the histopathological features. Patients with advanced fibrosis had higher serum Pin1 levels than the mild fibrosis group (53.42 ± 33.8 vs. 33.24 ± 20.90, respectively; p < 0.001). In multivariate analysis, Pin1 remained an independent predicting factor of advanced liver fibrosis (OR: 1.051, 95% CI: 1.013-1.089, p < 0.05). Conclusion: Serum Pin1 level can be used as a potential independent marker of the presence of the NASH and advanced fibrotic scores.

Sa1688 Investigation of the Relationship Between ACE I/D Gen Polymorphism and the Degree of Liver Fibrosis Due to Various Underlying Etiologies

Background and aim: Angiotensin 2, an important byproduct of renin angiotensin system can lead to increased extracellular matrix formation and fibrosis via enhanced TGF β-1 production in many organ systems including liver. Angiotensin converting enzyme (ACE) coding gen polymorphisms can lead to different degree of ACE expressions and hence, various angiotensin 2 tissue levels. ACE gen polymorphisms have been reported as ACE I/ D. Especially, ACE gen D homozygotic patients produce more tissue levels of ACE and this may be a potential hazard for many organ systems. We aimed to investigate if there is any role of ACE I/D gen polymorphism in the degree of liver fibrosis due to various etiologies. Methods: We enrolled 411 patients with a histopathological diagnosis of liver fibrosis. There were 240 patients with the diagnosis of mild and moderate fibrosis (Ishak's stage; 1-3) and the rest, 171 patients had advanced liver fibrosis (Ishak's stage; 4-6). Polymerase chain reaction was used to determine the type of ACE I/D gen polymorphisms. We also studied serum ACE levels with enzymatic kinetic testing method on spectrometry. Results: There were 208 male and 203 female subjects with a mean age of 51.1±13.5 years, (range; 1880 years). Within the mild and moderate liver fibrosis group, the etiologies were chronic viral hepatitis B in 131, chronic viral hepatitis C in 59, non-alcoholic steatohepatitis in 46, autoimmune hepatitis in 1 and primary biliary cirrhosis in 3 patients. Within the advanced liver fibrosis group, the etiologies were chronic viral hepatitis B in 71, chronic viral hepatitis C in 61, non-alcoholic steatohepatitis in 24, alcoholic liver disease in 11 and autoimmune hepatitis in 4 patients. On statistical analysis, there was no difference between the two groups with regard to ACE genotypes. Moreover, we could not find any significant difference between the groups regarding serum levels of angiotensin converting enzyme. Conclusion: The ACE I/D gen polymorphism do not seem to contribute as a significant risk factor for advanced liver fibrosis.

Sa1689 Microtubule Based Motility of Autophagic and Lysosomal Compartments In Vitro: Vesicles With LC3 on Their Surface Show Greater Motility Than Those That Contain Lamp1

Background and aim: Angiotensin 2, an important byproduct of renin angiotensin system can lead to increased extracellular matrix formation and fibrosis via enhanced TGF β-1 production in many organ systems including liver. Angiotensin converting enzyme (ACE) coding gen polymorphisms can lead to different degree of ACE expressions and hence, various angiotensin 2 tissue levels. ACE gen polymorphisms have been reported as ACE I/ D. Especially, ACE gen D homozygotic patients produce more tissue levels of ACE and this may be a potential hazard for many organ systems. We aimed to investigate if there is any role of ACE I/D gen polymorphism in the degree of liver fibrosis due to various etiologies. Methods: We enrolled 411 patients with a histopathological diagnosis of liver fibrosis. There were 240 patients with the diagnosis of mild and moderate fibrosis (Ishak's stage; 1-3) and the rest, 171 patients had advanced liver fibrosis (Ishak's stage; 4-6). Polymerase chain reaction was used to determine the type of ACE I/D gen polymorphisms. We also studied serum ACE levels with enzymatic kinetic testing method on spectrometry. Results: There were 208 male and 203 female subjects with a mean age of 51.1±13.5 years, (range; 1880 years). Within the mild and moderate liver fibrosis group, the etiologies were chronic viral hepatitis B in 131, chronic viral hepatitis C in 59, non-alcoholic steatohepatitis in 46, autoimmune hepatitis in 1 and primary biliary cirrhosis in 3 patients. Within the advanced liver fibrosis group, the etiologies were chronic viral hepatitis B in 71, chronic viral hepatitis C in 61, non-alcoholic steatohepatitis in 24, alcoholic liver disease in 11 and autoimmune hepatitis in 4 patients. On statistical analysis, there was no difference between the two groups with regard to ACE genotypes. Moreover, we could not find any significant difference between the groups regarding serum levels of angiotensin converting enzyme. Conclusion: The ACE I/D gen polymorphism do not seem to contribute as a significant risk factor for advanced liver fibrosis.

Sa1692 BVES Is Critical for Crowding Induced Cancer Cell Extrusion in Liver Cancer Metastasis

Background and aim: Angiotensin 2, an important byproduct of renin angiotensin system can lead to increased extracellular matrix formation and fibrosis via enhanced TGF β-1 production in many organ systems including liver. Angiotensin converting enzyme (ACE) coding gen polymorphisms can lead to different degree of ACE expressions and hence, various angiotensin 2 tissue levels. ACE gen polymorphisms have been reported as ACE I/ D. Especially, ACE gen D homozygotic patients produce more tissue levels of ACE and this may be a potential hazard for many organ systems. We aimed to investigate if there is any role of ACE I/D gen polymorphism in the degree of liver fibrosis due to various etiologies. Methods: We enrolled 411 patients with a histopathological diagnosis of liver fibrosis. There were 240 patients with the diagnosis of mild and moderate fibrosis (Ishak's stage; 1-3) and the rest, 171 patients had advanced liver fibrosis (Ishak's stage; 4-6). Polymerase chain reaction was used to determine the type of ACE I/D gen polymorphisms. We also studied serum ACE levels with enzymatic kinetic testing method on spectrometry. Results: There were 208 male and 203 female subjects with a mean age of 51.1±13.5 years, (range; 1880 years). Within the mild and moderate liver fibrosis group, the etiologies were chronic viral hepatitis B in 131, chronic viral hepatitis C in 59, non-alcoholic steatohepatitis in 46, autoimmune hepatitis in 1 and primary biliary cirrhosis in 3 patients. Within the advanced liver fibrosis group, the etiologies were chronic viral hepatitis B in 71, chronic viral hepatitis C in 61, non-alcoholic steatohepatitis in 24, alcoholic liver disease in 11 and autoimmune hepatitis in 4 patients. On statistical analysis, there was no difference between the two groups with regard to ACE genotypes. Moreover, we could not find any significant difference between the groups regarding serum levels of angiotensin converting enzyme. Conclusion: The ACE I/D gen polymorphism do not seem to contribute as a significant risk factor for advanced liver fibrosis.