Histological Fibrosis Stage Research Articles

Simpler Novel Non-Invasive Program Score for Stage-by-Stage Diagnosis of Liver Fibrosis in Untreated Patients with Chronic Hepatitis B: A Multicenter Prospective Study

Background: Non-invasive evaluation for liver fibrosis is of great clinical interest and value, but current models fail to accurately stage and differentiate each stage of liver fibrosis, especially in patients with chronic hepatitis B (CHB), and never taking off to be used clinically. Methods: This multicenter, prospective study used unbiased penalized logistic regression, assessed 30 variables referencing to histologic fibrosis stage in a large training cohort (n=800), and established simpler novel non-invasive program score (SNNPS). An independent validation cohort (n=400) was used to validate the SNNPS. Finally, a mobile program was then developed for one-step detailed calculation of certain hepatic fibrosis staging. Findings: Five variables-liver stiffness measurement (LSM), platelet counts, age, serum hyaluronic acid and spleen diameter, were identified as independent predictors for fibrosis stage and developing SNNPS that has AUCs of 0.893, 0.897, and 0.909 for significant fibrosis, advanced fibrosis, and cirrhosis, respectively. Using sub-models of SNNPS, S1 score ≤ 2.875 was 86% specific and 82% sensitive for mild fibrosis, S2 score ≤ 4.06 was 85% specific and 86% sensitive for significant fibrosis, and S3 score ≤ 4.402 or > 4.402 was 93% specific and 91% sensitive for advanced fibrosis or cirrhosis. In validation set, AUCs for significant fibrosis, advanced fibrosis, and cirrhosis were 0.904, 0.912, and 0.897, respectively. The SNNPS with highest LR and lowest LR- was significantly more specific and sensitive than AAR, APRI, LSM alone and Hepascore, and can be calculated via a mobile program in one-step. Interpretation: Using sophisticated mathematic modelling and a large multicenter study data developed and validated SNNPS that is superior to previously reported models, and can be easily calculated via mobile program to accurately diagnose various hepatic fibrosis stages in CHB patients in a wider range. Trial Registration Number: ClinicalTrials.gov. Identifier: NCT65418 Funding Information: National Major Science and Technology Special Project of China (2013ZX10005002). Competing Interest Declaration: The authors declare that there is no conflict of interest in this study Ethical Approval Statement: The study meets the requirements of Declaration of Helsinki, and protocol was approved by the ethics committee of each participating institution, and a written informed consent was obtained from all patients.

Senescence mirrors the extent of liver fibrosis in chronic hepatitis C virus infection.

Chronic viral hepatitis is linked to fibrotic liver injury that can progress to liver cirrhosis with its associated complications. Recent evidence suggests a role of senescence in liver fibrosis, although the senescence regulators contributing to fibrosis progression remain unclear. To investigate the role of senescence and different senescence markers for fibrosis progression in patients with chronic hepatitis C virus (HCV) infection. The expression of the cell cycle inhibitors p21, p27 and p16 as well as the senescence markers p-HP1γ and γ-H2AX was analysed in liver tissue with different fibrosis stages. Senescence-associated chitotriosidase activity was measured in sera of HCV patients (n=61) and age-matched healthy individuals (n=22). We found a remarkable up-regulation of the cell cycle inhibitors and senescence markers in chronic HCV infection compared to healthy liver tissue. Liver tissue with relevant fibrosis stages (F2-3) or cirrhosis (F4) revealed a significant increase in senescent cells compared to livers with no or minimal fibrosis (F0-1). In cirrhotic livers, a significantly higher number of p-HP1γ, p21 and p27 positive cells was detected compared to liver tissue with F2-3 fibrosis. Importantly, we identified T-cells as the dominant cell type contributing to increased senescence during fibrosis progression. Compared to healthy individuals, serum chitotriosidase was significantly elevated and correlated with histological fibrosis stages and liver stiffness as assessed by transient elastography. Senescence of hepatic T-cells is enhanced in chronic viral hepatitis and increases with fibrosis progression. Serological detection of senescence-associated chitotriosidase might allow for the non-invasive detection of relevant fibrosis stages.

Non-Invasive Assessment of Liver Fibrosis in a Rat Model: Comparison of Shear Wave Elastography with Serologic Tests

Background: Non-invasive evaluation of hepatic fibrosis, especially serum biomarkers and ultrasound elastography, has vigorously developed during the past decades. Objectives: The purpose of this study was to evaluate the diagnostic performance of the latest generation of ultrasound elastography- two-dimensional real-time shear wave elastography (SWE) in accessing liver fibrosis progression in comparison with serologic tests in a rat model. Materials and Methods: Liver fibrosis was induced in 108 rats (control group: 18 rats, experiment group: 90 rats) by intraperitoneal injection of thioacetamide (300mg/kg, twice a week for 12 weeks). SWE examination was done by two operators. Hyaluronic acid (HA), aspartate aminotransferase (AST) to platelet (PLT) ratio index (APRI), AST to alanine aminotransferase (ALT) ratio (AAR) were used for serology tests. Histological fibrosis stages (F0-F4) according to the Scheuer scoring system were referred as the gold standard. Diagnostic performance of elastography and serology methods were determined by the areas under receiver operating characteristic curve (AUROC). Results: Liver stiffness values increased with the progression of hepatic fibrosis. Liver stiffness assessed by SWE exhibited a strong positive correlation with histological fibrosis stages (r = 0.849, P < 0.001). SWE showed a better diagnostic performance than the selected serological biomarkers HA, APRI, and AAR (AUROC for SWE, HA, APRI, and AAR were 0.933, 0.707, 0.714, and 0.691 in diagnosing F ≥ F2 and 0.986, 0.807, 0.870, and 0.770 in diagnosing F = F4, respectively). Conclusion: Ultrasound elastography SWE technique is a reliable method for noninvasive evaluation of liver fibrosis in comparison with the serological biomarkers, HA, APRI, and AAR.

The combination of 5 serum markers compared to FibroScan to predict significant liver fibrosis in patients with chronic hepatitis B virus

BackgroundWe evaluated the performance of serum hyaluronan (HA), procollagen type III N-terminal peptide (PIIINP), type IV collagen (IVC), laminin (LN), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), compared to transient elastography (FibroScan) in predicting significant liver fibrosis. MethodsWe therefore determined 4 serum fibrosis markers, FibroScan and liver biopsy in 70 consecutive adult patients with chronic hepatitis B. According to a modified Scheuer scoring system, significant fibrosis was defined as fibrosis stage ≥S2. We compared serum fibrosis markers to histological staging and FibroScan results using Spearman correlation analysis and area under receiver operating characteristic (ROC) curves (AUROCs). ResultsOf the 212 patients who had the results of FibroScans and four serum fibrosis markers for HBV, 70 had concurrent liver biopsy. Significant liver fibrosis was found in 24/70 patients. The serum levels of HA, PIIINP, IVC, LN, ALT, AST was all positively correlated with fibrosis stage of Liver biopsy. The coefficients with stages were respectively 0.468, 0.392, 0.538, 0.213, 0.350, 0.375. There was a significant difference between mild fibrosis (<S2) and significant fibrosis (≥S2), excluding LN, in the levels of these 5 serum makers (P < .05). AUROC for FibroScans and HA, PIIINP, IVC, LN, ALT, AST to correctly allocate patients to histological fibrosis stage ≥ S2 was 0.866, 0.784, 0.738, 0.827, 0.630, 0.713 and 0.728 respectively. Since LN shows the worst performance of the others. We decided to check the performance of the combination of HA, PIIINP, CIV, ALT, AST, excluding LN, to distinguish fibrosis stages. The index of the histological fibrosis stage ≥ S2, combining the 5 serum markers, significantly improved diagnostic performance (AUROC = 0.861) compared to the use of 5 serum markers alone in all HBV patients. ConclusionThe combination of the 5 serum markers and FibroScan performed equally well in predicting significant fibrosis. The combination of the 5 serum markers is a reliable noninvasive method to predict significant liver fibrosis in patients with CHB. So, it provide another choice rather than FibroScan in predicting significant liver fibrosis.

Chemokine Receptor 5 Has No Major Role in the Severity of Hepatitis C Virus-Related Liver Damage.

Total or partial inactivation of the chemokine 5 (CC5) pathway, as caused by the CC5 receptor Δ32 deletion (CCR5Δ32), may result in a profound manipulation of immune surveillance with significant consequences on the course and response to therapy of diverse human infections, including HIV. It has been postulated that in chronic hepatitis C (CHC), such a deregulation of CC5 pathway may compromise T cell-dependent antiviral immune responses, which in turn may favor viral persistence. To test this hypothesis, we investigated a cohort of 100 patients with CHC in whom 12 heterozygous and 1 homozygous CCR5Δ32 mutations were detected compared to 8 and none in 98 healthy controls (13% vs. 8.2%, p = 0.36). As patients with and without CCR5Δ32 mutations were similar in terms of histological activity (p = 0.84) and fibrosis stage (p = 0.20) as well as CCR5 tissue expression, we reasonably exclude that this CCR5 mutation is significantly involved in the pathogenesis of CHC and may be a potential therapeutic target. However, deleted patients showed a significantly higher response to pegylated interferon-alfa (PEG-IFN), suggesting that a dormant immune system is more readily primed by immunostimulation.

Anti-fibrotic role and mechanism of Periplaneta americana extracts in CCl4-induced hepatic fibrosis in rats.

Hepatic fibrosis is resulted from sustained wound-healing responses to various harmful stimuli, including viral infection, drug toxicity, alcohol, and autoimmune hepatopathy, and it has recently attracted the attention of an increasing number of researchers and clinical workers. The aims of this study were to examine the anti-fibrotic effects of extracts of Periplaneta americana (EPA) on CCl4-induced hepatic fibrosis in rats, to preliminary determine the anti-fibrotic efficacy of EPA, and to identify a potential and effective therapeutic agent to attenuate hepatic fibrosis. In this study, we routinely detected liver functional indices, such as alanine aminotransferase (ALT), aspartate transaminase (AST), and albumin (Alb). We also measured hepatic fibrosis-related serum markers, including hyaluronic acid (HA), laminin (LN), type III procollagen (PC III), and type IV collagen (IV-C) via radioimmunoassay. Moreover, we examined histological activity and fibrosis stage via light microscopy after hematoxylin and eosin and Masson staining. Furthermore, we detected the expressions of nuclear factor-kappa B (NF-κB), alpha-smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), and tissue inhibitor of metalloprotease-1 (TIMP-1) in rat liver tissues by immunohistochemical staining. We found that EPA, whose main components are viscous sugar amino acids, can reduce the levels hepatic fibrosis-related factors, including HA, LN, PC III, and IV-C, improve liver function, attenuate, or reverse pathological damage associated with hepatic fibrosis, and thus inhibit the progression of hepatic fibrosis. The mechanism of EPA action may be related to the inhibition of TGF-β1, NF-κB, and α-SMA expressions and the reduction of TIMP-1 levels in the liver to reduce the accumulation of extracellular matrix (ECM) components, thereby blocking the relevant signaling pathways and preventing inflammatory responses to attenuate or reverse hepatic fibrosis. EPA may thus be used as a potentially effective therapeutic agent for the treatment of hepatic fibrosis.

Evaluation of transient elastography in assessing liver fibrosis in patients with advanced schistosomiasis japonica

Schistosomiasis remains an important public health issue. The presence and extent of liver fibrosis are associated with disease progression and prognosis. The study is aimed at exploring the value of liver stiffness measurement (LSM) by transient elastography in assessing liver fibrosis in patients with advanced schistosomiasis japonica. Seventy-three patients were consecutively recruited for the purpose of this study. The correlation between noninvasive parameters and histological fibrosis stages was analyzed and an area under receiver operating characteristic curve (AUROC) was used to assess diagnostic efficacy. Our results demonstrated that there are significant differences between LSM values of patients with different stages of fibrosis (F1 vs. F2, F2 vs. F3 and F3 vs. F4, P<0.01). The AUROC values of LSM in detecting significant fibrosis (F≥2), advanced fibrosis (F≥3) and cirrhosis (F=4) were 0.96, 0.90, and 0.92 respectively. The optimal cut-off LSM values were 8.0kPa, 9.5kPa, and 18.0kPa for significant fibrosis, advanced fibrosis and cirrhosis. Based on differences between AUROC values, LSM was proven to be superior to several serum models in detecting advanced fibrosis and cirrhosis. In conclusion, our study demonstrates that LSM is a reliable parameter for assessing risk of liver fibrosis in patients with advanced schistosomiasis japonica.

Incidentally detected asymptomatic hepatitis C virus infection with significant fibrosis: Possible impacts on management.

Liver biopsy may be considered in patients with hepatitis C virus (HCV) infection to assess the severity of liver injury and stage of fibrosis, thereby guiding therapeutic decisions. In addition, advanced stage also necessitates surveillance for hepatocellular carcinoma. The aim of this study was to assess whether transaminase (alanine transaminase [ALT]) levels and RNA titers correlate with the histological activity index (HAI) and fibrosis (F) stage in asymptomatic patients with incidentally detected HCV (IDHCV). Retrospective evaluation of liver biopsies was done in 113 patients with IDHCV, diagnosed during routine screening. Decision of liver biopsy was made on the basis of age, genotype, acceptable clinical, hematological, and biochemical profiles, and willingness of the patients to undergo treatment. Serum ALT levels, HCV RNA titers, and genotypes were correlated with HAI and F stage. Genotyping was done in 77 of the 113 patients, of which genotype 3 was seen in 43 and genotype 1 in 25 patients. A higher fibrosis stage (Ishak's >F2) was noted in 23.8% of the biopsies. Serum ALT showed a significant correlation with the HAI score on liver biopsy (P = 0.01) but not with the stage of fibrosis (P = 0.52). HCV RNA titers did not reveal any correlation with HAI score or fibrosis stage. Serum transaminases and HCV RNA titers are poor predictors of disease severity and fibrosis. Since HCV shows a slow disease progression, higher stage may predict a worse prognosis irrespective of the low viral RNA load. Liver biopsy may help guide therapeutic decisions in IDHCV infection.

Utility and cost evaluation of multiparametric magnetic resonance imaging for the assessment of non-alcoholic fatty liver disease.

Validated diagnostic tools that are accurate, cost effective and acceptable to patients are required for disease stratification and monitoring in NAFLD. To investigate the performance and cost of multiparametric MRI alongside existing biomarkers in the assessment of NAFLD. Adult patients undergoing standard of care liver biopsy for NAFLD were prospectively recruited at two UK liver centres and underwent multiparametric MRI, blood sampling and transient elastography withing 2weeks of liver biopsy. Non-invasive markers were compared to histology as the gold standard. Data were obtained in 50 patients and 6 healthy volunteers. Corrected T1 (cT1) correlated with NAFLD activity score (ρ=0.514, P<.001). cT1, enhanced liver fibrosis (ELF) test and liver stiffness differentiated patients with simple steatosis and NASH with AUROC (95% CI) of 0.69 (0.50-0.88), 0.87 (0.77-0.79) and 0.82 (0.70-0.94) respectively and healthy volunteers from patients with AUROC (95% CI) of 0.93 (0.86-1.00), 0.81 (0.69-0.92) and 0.89 (0.77-1.00) respectively. For the risk stratification of NAFLD, multiparametric MRI could save £150,218 per 1000 patients compared to biopsy. Multiparametric MRI did not discriminate between individual histological fibrosis stages in this population (P=.068). Multiparametric MRI accurately identified patients with steatosis, stratifies those with NASH or simple steatosis and reliably excludes clinically significant liver disease with superior negative predictive value (83.3%) to liver stiffness (42.9%) and ELF (57.1%). For the risk stratification of NAFLD, multiparametric MRI was cost effective and, combined with transient elastography, had the lowest cost per correct diagnosis.

Quantitative assessment of liver fibrosis reveals a nonlinear association with fibrosis stage in nonalcoholic fatty liver disease.

Accurate staging of liver fibrosis is crucial to guide therapeutic decisions for patients with nonalcoholic fatty liver disease (NAFLD). Digital image analysis has emerged as a promising tool for quantitative assessment of fibrosis in chronic liver diseases. We sought to determine the relationship of histologic fibrosis stage with fiber amounts quantified in liver biopsy specimens for the better understanding of NAFLD progression. We measured area ratios of collagen and elastin fibers in Elastica van Gieson‐stained biopsy tissues from 289 patients with NAFLD from four hospitals using an automated computational method and examined their correlations with Brunt's fibrosis stage. As a secondary analysis, we performed multivariable logistic regression analysis to assess the associations of the combined area ratios of collagen and elastin with noninvasive fibrosis markers. The combined fiber area ratios correlated strongly with Brunt's stage (Spearman correlation coefficient, 0.78; P < 0.0001), but this relationship was nonlinear (P = 0.007) with striking differences between stage 4 (median area ratios, 12.3%) and stages 0‐3 (2.1%, 2.8%, 4.3%, and 4.8%, respectively). Elastin accumulation was common in areas of thick bridging fibrosis and thickened venous walls but not in areas of perisinusoidal fibrosis. The highest tertile of the combined fiber area ratios was associated with the fibrosis‐4 index and serum type IV collagen 7s domain (7s collagen) levels, whereas the upper two tertiles of the fiber amounts significantly associated with body mass index, aspartate aminotransferase, and 7s collagen in the multivariable analysis. Conclusion: Quantitative fibrosis assessment reveals a nonlinear relationship between fibrosis stage and fiber amount, with a marked difference between stage 4 and stage 3 and much smaller differences among stages 0‐3, suggesting a heterogeneity in disease severity within NAFLD‐related cirrhosis. (Hepatology Communications 2018;2:58–68)

The non-invasive serum biomarker soluble Axl accurately detects advanced liver fibrosis and cirrhosis

Soluble Axl (sAxl) was recently shown to be strongly released into the blood during liver fibrogenesis and hepatocellular carcinoma suggesting sAxl as a biomarker of liver diseases. In this study we are the first to evaluate sAxl in human serum in comparison to Enhanced Liver Fibrosis (ELF) test and transient elastography (TE; Fibroscan) for its value to detect significant (F≥2), advanced fibrosis (F≥3), and cirrhosis (F4) in different liver disease etiologies and healthy controls. To properly determine the diagnostic accuracy of sAxl, a test cohort as well as a validation cohort was employed using liver biopsy as a reference method. Most notably, sAxl was confirmed to be an accurate biomarker of liver fibrosis and cirrhosis. Its accuracy was increased, if total serum albumin was added to build a sAxl/albumin ratio. Thereby an AUC of 0.763, 0.776, 0.826, and 0.832 was achieved corresponding to histological fibrosis stages F≥2, F≥3, F4 with liver biopsy as a reference method, and cirrhosis according to imaging techniques, respectively. With a cut-off of 1.29, a sensitivity, specificity, PPV, and NPV of 78.5%, 80.1%, 44%, 94.9% for the detection of cirrhosis was achieved. In comparison, ELF test and TE showed an AUC of 0.910, and 0.934, respectively, for the detection of cirrhosis. However, performance of TE was not possible in 14.4% of patients and both, ELF™ test and TE bear the disadvantage of high costs. In conclusion, the sAxl/albumin ratio is suggested as an accurate biomarker of liver fibrosis and cirrhosis. Due to its easy applicability and low costs it is suitable as screening parameter for significant to advanced liver fibrosis and cirrhosis, especially if TE is not available or not applicable.

Diagnostic accuracy of the Coopscore© to predict liver fibrosis in human immunodeficiency virus/hepatitis B virus co-infection.

Background Non-invasive methods for assessing liver fibrosis are increasingly used as an alternative to liver biopsy. Recently, a score-based biochemical blood test (Coopscore©) was developed in a cohort of patients chronically infected with hepatitis C virus, showing higher diagnostic performances than Fibrometer®, Fibrotest®, Hepascore® and Fibroscan™. Here, we assess its performance in patients co-infected with the human immunodeficiency virus and hepatitis B virus. Methods Ninety-seven human immunodeficiency virus/hepatitis B virus co-infected patients with liver biopsies were included from a previously described cohort. Histological fibrosis staging using METAVIR criteria was used as the reference. Coopscore©, Fibrotest®, Fibrometer®, Hepascore® and Zeng score were computed and compared with the Coopscore© using the Obuchowski index and area under the receiving operator characteristic curves. Results The distribution of liver fibrosis levels was as follows: F0-F1 ( n = 42), F2 ( n = 25), F3 ( n = 15) and F4 ( n = 15). The Obuchowski index was higher for Coopscore© (0.774) than Fibrometer® (0.668), Hepascore® (0.690) and Zeng scores (0.704) ( P < 0.05), reflecting a better ability to discriminate between fibrosis stages. Similarly, when predicting significant fibrosis (≥F2), the AUROC was significantly greater for the Coopscore© (0.836) than the Hepascore® (0.727) and Zeng scores (0.746), but not for the Fibrotest® (0.778, P = 0.14) or Fibrometer® (0.790, P = 0.19). The Coopscore© did not show a higher capacity than other scores to predict advanced fibrosis (≥F3) or cirrhosis (F4). Conclusions This study supports the diagnostic value of the Coospcore© in fibrosis staging among human immunodeficiency virus/hepatitis B virus co-infected patients, especially to predict significant fibrosis.

Real-time elastography (RTE): a valuable sonography-based non-invasive method for the assessment of liver fibrosis in chronic hepatitis B.

To investigate the diagnostic usefulness of real-time elastography (RTE) for liver fibrosis in chronic hepatitis B (CHB). 89 CHB patients were enrolled in the cross-sectional study. Ultrasound-guided percutaneous liver biopsies, RTE, and blood testing were performed in all patients. Areas under receiver operating characteristic curves (AUROC) were used to examine the diagnostic performance of liver fibrosis index (LFI) for the assessment of liver fibrosis. LFI differed significantly across histologic fibrosis stages (P<0.05), except the comparison between S0 and S1 (P=0.298). There was a strong positive correlation between LFI and histologic liver fibrosis stage (Spearman r=0.831, P<0.001). The cutoff LFI value of >2.74 indicated a sensitivity of 0.766 and a specificity of 0.872 for predicting significant liver fibrosis (S≥2), and the cutoff LFI value of >3.61 indicated a sensitivity of 0.833 and a specificity of 0.878 for predicting early liver cirrhosis (S=4). LFI showed higher AUROC for discriminating significant liver fibrosis (0.873 vs. 0.614) and early liver cirrhosis (0.923 vs. 0.769) than aspartate aminotransferase-to-platelet ratio index (APRI). RTE is a valuable sonography-based non-invasive method for assessment of liver fibrosisand has better discrimination power for significant liver fibrosis and early liver cirrhosis than APRI in CHB.

A randomised controlled trial of losartan as an anti-fibrotic agent in non-alcoholic steatohepatitis.

IntroductionNon-alcoholic fatty liver disease (NAFLD) is a common liver disease worldwide. Experimental and small clinical trials have demonstrated that angiotensin II blockers (ARB) may be anti-fibrotic in the liver. The aim of this randomised controlled trial was to assess whether treatment with Losartan for 96 weeks slowed, halted or reversed the progression of fibrosis in patients with non-alcoholic steatohepatitis (NASH).MethodsDouble-blind randomised-controlled trial of Losartan 50 mg once a day versus placebo for 96 weeks in patients with histological evidence of NASH. The primary outcome for the study was change in histological fibrosis stage from pre-treatment to end-of-treatment.ResultsThe study planned to recruit 214 patients. However, recruitment was slower than expected, and after 45 patients were randomised (median age 55; 56% male; 60% diabetic; median fibrosis stage 2), enrolment was suspended. Thirty-two patients (15 losartan and 17 placebo) completed follow up period: one patient (6.7%) treated with losartan and 4 patients (23.5%) in the placebo group were “responders” (lower fibrosis stage at follow up compared with baseline). The major reason for slow recruitment was that 39% of potentially eligible patients were already taking an ARB or angiotensin converting enzyme inhibitor (ACEI), and 15% were taking other prohibited medications.ConclusionsDue to the widespread use of ACEI and ARB in patients with NASH this trial failed to recruit sufficient patients to determine whether losartan has anti-fibrotic effects in the liver.Trial registrationISRCTN 57849521

Diagnostic value of Transient Elastography (Fibroscan) in the evaluation of liver fibrosis in chronic viral hepatitis C: Comparison to liver biopsy

PurposeTo assess the Transient Elastography diagnostic and staging role in liver fibrosis in chronic hepatitis C in comparison to hepatic biopsy. Material and methods70 chronic viral hepatitis C patients who are candidate for therapy using antiviral drugs underwent Transient Elastography and percutaneous hepatic biopsy on the same day. Measurements of liver stiffness obtained by Fibroscan were correlated with different histological stages of fibrosis. The diagnostic yield of the measurements obtained by Fibroscan were compared to histolopathological stage of fibrosis (Metavir score) to achieve the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) as well as Kappa test. Receiver operating characteristic (ROC) curves were analyzed. ResultsThere was a correlation between Fibroscan measurements and histological fibrosis stage (r=0.87, p<0.001). The Area under the curve (AUC for detection of fibrosis (≥F1) was 0.91 with a sensitivity, specificity, PPV and NPV of 88%, 77.8%, 96.4% and 50.0% respectively while kappa test and p-value were 0.53 and <0.001 respectively at cut off value of 4.95kPa. The AUC for fibrosis (≥F2) was 0.93 with a sensitivity, specificity, PPV, NPV, kappa test and p-value 92.0%, 65.7%, 76.1%, 87.5%, 0.59 and <0.001 respectively at cut off value of 6.25kPa. The AUC for sever fibrosis (≥F3) was 0.97 with a sensitivity, specificity, PPV, NPP, kappa test and p-value 95%, 89.2%, 80.0%, 97.5%, 0.80 and <0.001 respectively at cut off value of 10.1kPa. For cirrhosis (=F4) the AUC was 0.98 with a sensitivity, specificity, PPV, NPV, kappa test and p-value 91.7%, 98.3%, 91.7%, 98.5%, 0.89 and <0.001 respectively at cut off value of 17.15kPa. ConclusionTransient Elastography is a useful diagnostic method with significantly higher correlation with the liver fibrosis histological grade which is a crucial factor in proper management.

Serum ferritin concentration predicts hepatic fibrosis better than hepatic iron concentration in human HFE-Haemochromatosis.

Ferritin is purported to have proinflammatory and profibrogenic effects on hepatic stellate cells. Thus, rather than acting as a passive indicator of hepatic iron concentration (HIC) in haemochromatosis, ferritin may directly influence fibrosis. This study evaluated whether serum ferritin is a better predictor of hepatic fibrosis compared to variables previously associated with increased fibrosis risk in haemochromatosis. We identified 291 C282Y HFE-homozygous patients who had undergone liver biopsy for histological fibrosis staging and measurement of HIC. Ordinal logistic regression determined the best model for fibrosis stage not including serum ferritin. Then, serum ferritin was introduced into this model to assess whether the predictive power of the model was significantly increased and to evaluate the effect on other predictors of fibrosis. Ordinal logistic regression analyses without serum ferritin demonstrated that log HIC (OR 2.89; P<.001), male gender (OR 2.93; P=.005), alcohol consumption (g/day) (OR 1.01; P=.004), steatosis (OR 2.86; P=.01), arthritis (OR 2.46; P=.01) predicted increasing fibrosis stage (n=217). Addition of serum ferritin in multivariate analysis substantially improved the predictive power of the model (χ2 =37.15; P<.01) and was highly predictive of fibrosis stage (OR 5.44; P<.001). Inclusion of serum ferritin in this model rendered the effects of HIC, gender, alcohol and steatosis to non-significance. In haemochromatosis, serum ferritin is a better predictor of fibrosis stage than HIC, gender, steatosis and alcohol. These data support a hypothesis that ferritin may play a role in fibrosis rather than simply acting as a passive indicator of iron storage.

Expression of Hepatocyte Hepatitis B Core Antigen and Hepatitis B Surface Antigen as a Marker in the Management of Chronic Hepatitis B Patients.

Background/AimsWe aimed to clarify the association of hepatitis B surface antigen (HBsAg)/hepatitis B core antigen (HBcAg) with the disease status and treatment response in patients with chronic hepatitis B (CHB).MethodsWe investigated 171 biopsy-proven entecavir-treated CHB patients (109 hepatitis B e antigen [HBeAg]-positive, 62 HBeAg-negative). HBcAg expression was positive when ≥10% of hepatocytes stained, and classified into nuclear, mixed, and cytoplasmic patterns. HBsAg expressions were intracytoplasmic (diffuse, globular, and submembranous) and membranous. The histologic activity index (HAI) and fibrosis stage followed Ishak system.ResultsIn HBeAg-positive patients, older age, increased HAI score, advanced fibrosis, and reduced viral load were observed when HBcAg expression shifted from nucleus to cytoplasm in HBcAg-positive patients, and HBsAg expression from non-submembranous to submembranous in HBcAg-negative patients (all, p<0.05). In HBeAg-negative patients, only intracytoplasmic HBsAg expression patterns had clinical relevance with decreased ALT levels and viremia. In HBeAg-positive patients without favorable predictors of virologic response, negative HBcAg and membranous HB-sAg expression predicted greater virologic response (both, p<0.05). The probability of HBeAg seroclearance was higher in patients with increased HAI or lacking HBcAg expression (both, p<0.05). Higher serum HBsAg levels and hepatocyte HBcAg positivity were associated with reduced serum HBsAg during first and post-first year treatment, respectively (both, p<0.05).ConclusionsHepatocyte HBcAg/HBsAg expression is a good marker for disease status and predicting treatment response.

Epigenetically-Regulated MicroRNA-9-5p Suppresses the Activation of Hepatic Stellate Cells via TGFBR1 and TGFBR2

Background/Aims: Recently, microRNAs (miRNAs) have been demonstrated to act as regulators of activation of hepatic stellate cells (HSCs). It is well known that the main profibrogenic inducer transforming growth factor-β1 (TGF-β1) contributes to HSC activation, which is a key event in liver fibrosis. Increasing studies show that miR-9-5p is down-regulated in liver fibrosis and restoration of miR-9-5p limits HSC activation. However, the role of miR-9-5p in TGF-β1-induced HSC activation is still not clear. Methods: miR-9-5p expression was quantified using real-time PCR in chronic hepatitis B (CHB) patients and TGF-β1-treated LX-2 cells. In CHB patients, histological activity index (HAI) and fibrosis stages were assessed using the Ishak scoring system. Effects of miR-9-5p on liver fibrosis in vivo and in vitro were analyzed. Luciferase activity assays were performed to examine the binding of miR-9-5p to the 3′-untranslated region of type I TGF-β receptor (TGFBR1) as well as TGFBR2. Results: Compared with healthy controls, miR-9-5p was reduced in CHB patients. There was a lower miR-9-5p expression in CHB patients with higher fibrosis scores or HAI scores. miR-9-5p was down-regulated by TGF-β1 in a dose-dependent manner. TGF-β1-induced HSC activation including cell proliferation, α-SMA and collagen expression was blocked down by miR-9-5p. Notably, miR-9-5p ameliorates carbon tetrachloride-induced liver fibrosis. As determined by luciferase activity assays, TGFBR1 and TGFBR2 were targets of miR-9-5p. Further studies demonstrated that miR-9-5p inhibited TGF-β1/Smads pathway via TGFBR1 and TGFBR2. Interestingly, promoter methylation was responsible for miR-9-5p down-regulation in liver fibrosis. The relationship between miR-9-5p expression and methylation was confirmed in CHB patients and TGF-β1-treated cells. Conclusion: Our results demonstrate that miR-9-5p could inhibit TGF-β1-induced HSC activation through TGFBR1 and TGFBR2. Loss of miR-9-5p is associated with its methylation status in liver fibrosis.

Prevalence and clinical implications of IL28B genotypes in Taiwanese patients with chronic hepatitis C.

Clinical implications of IL28B gene in Taiwanese chronic hepatitis C (CHC) patients remain unknown. We thus investigated the prevalence and clinical implications of IL28B rs8099917 genotypes in CHC patients with different hepatitis C virus (HCV) genotypes and healthy controls. A total of 200 HCV genotype 1 patients and 200 HCV genotype 2 patients who received liver biopsy, as well as 197 healthy controls were enrolled to determine the frequencies of IL28B rs8099917 genotypes. In addition, the association of IL28B rs8099917 genotype with baseline data, including HCV RNA level, HCV genotype, histological activity grade, fibrosis stage, and body mass index, were evaluated and further stratified by covariant factors. Compared with healthy controls, CHC patients had a lower prevalence rate of favorable IL28B rs8099917 TT genotype (81.0% vs. 89.3%, p=0.025). In addition, the prevalence rates of favorable TT genotype in patients with HCV genotypes 1 and 2 were 76.0% and 86.0%, respectively (p=0.007). Using ordered logistic regression analysis, higher fibrosis stages were found to be associated with a lower prevalence of TT genotype (p=0.033), but not histological activity grades (p=0.748). The association with fibrosis stages was more pronounced in female patients (p=0.024). In Taiwan, CHC patients have a lower frequency of favorable IL28B TT genotype than healthy controls. Among patients with CHC, the frequency of TT genotype is higher in HCV genotype 2 patients than in HCV genotype 1 patients. In addition, CHC patients with TT genotype, particularly females, have a lower likelihood of advanced fibrosis.

Quantitative evaluation of pancreatic tumor fibrosis using shear wave elastography

Background & aimsThere is no established non-invasive method for diagnosis of pancreatic fibrosis. Shear wave elastography (SW-EG) may be a candidate for this purpose. The aims of this study were to assess the reproducibility of SW-EG in the normal imaging pancreas (Phase 1) and to evaluate the diagnostic performance of SW-EG for pancreatic fibrosis classified histologically (Phase 2). MethodsPhase 1: This included 127 cases that underwent SW-EG of the normal imaging pancreas. SW-EG was measured at least five times in the pancreatic parenchyma and the median of repeated measurements was defined as the pancreatic elastic modulus (PEM). Phase 2: This included 53 cases that underwent SW-EG of the pancreatic parenchyma preoperatively and in which pancreas parenchyma were evaluated histologically. Histological fibrosis was graded in 4 stages: normal, mild, moderate, and severe. ResultsPhase 1: Median PEM in the head, body, and tail of the pancreas were 3.23, 3.17, and 2.91 kPa, respectively, with no significant difference among regions (P = 0.554). The intraclass correlation coefficient showed good reproducibility (ρ = 0.71) after 5 measurements. Phase 2: There was a significant positive correlation between PEM and the histological pancreatic fibrosis stage (rs = 0.63, P < 0.001). Areas under the receiver operating characteristic curve for the accuracy of SW-EG for diagnosis of pancreatic fibrosis were 0.85 (≥mild), 0.84 (≥moderate), and 0.87 (severe). ConclusionSW-EG can be used to determine the stage of pancreatic fibrosis non-invasively with high accuracy and reproducibility.