Chronic Hepatitis Cirrhosis Research Articles

Research on the relationship of intrahepatic expression of IL-17 and liver cirrhosis in chronic hepatitis B patients

Research on the relationship of intrahepatic expression of IL-17 and liver cirrhosis in chronic hepatitis B patients

Liver Stiffness Measurement in Combination With Noninvasive Markers for the Improved Diagnosis of B-viral Liver Cirrhosis

To investigate the performance of liver stiffness measurement (LSM) in combination with available noninvasive markers in hepatitis B virus-related chronic liver disease. Few noninvasive methods are available for predicting liver cirrhosis in chronic hepatitis B (CHB). Between January 2006 and June 2007, we studied 130 consecutive treatment-naive CHB patients who underwent liver biopsy (LB) and LSM. The aspartate to alanine aminotransferase ratio, age-platelet index (API), aspartate aminotransferase to platelet ratio index (APRI), LSM, and their combinations were compared with liver histology. The API, APRI, and LSM, but not the aspartate to alanine aminotransferase ratio, correlated significantly with liver cirrhosis (all P<0.001). The diagnostic accuracy of LSM and API exceed that of the other diagnostic methods for predicting liver cirrhosis (area under the receiver operating characteristic curve=0.840 and 0.818). When LSM was combined with API and APRI, the diagnostic accuracy was improved markedly (area under the receiver operating characteristic curve =0.871, and 0.846). When both LSM and API results were in agreement, LB confirmed them in 89.1% (41/46) of cases for liver cirrhosis. LB could have been avoided in 41 (31.5%) of the 130 patients who were examined for the potential diagnosis of liver cirrhosis. The combination of LSM and API can avoid unnecessary invasive LB procedures in CHB patients.

Improved diagnostic accuracy of blood tests for severe fibrosis and cirrhosis in chronic hepatitis C

Blood tests are usually designed to identify significant fibrosis. We evaluated their diagnostic accuracy, and how to increase it, for the clinically important targets of severe fibrosis and cirrhosis. The accuracy for severe fibrosis or cirrhosis of four blood tests was evaluated based on Metavir staging in 1056 patients with chronic hepatitis C recruited in five independent hospitals. Using original scores, an original diagnostic target (significant fibrosis) and best diagnostic cutoff, the correct classification rates in severe fibrosis and cirrhosis stages were, respectively: FibroMeter: 90.1, 100%, Fibrotest: 78.2, 95.1%, Hepascore: 73.8, 94.9%, aspartate aminotransferase to platelet ratio index (APRI): 71.4, 88.0% (P<0.003, P=0.004, respectively, between tests). The corresponding area under the receiver operating characteristics were FibroMeter: 0.885, 0.907, Fibrotest: 0.837, 0.882, Hepascore: 0.834, 0.896, APRI: 0.822, 0.841 (P<0.003, respectively). Observed 100% negative predictive values for severe fibrosis and cirrhosis were, respectively, FibroMeter: 15.4, 47.5%, Fibrotest: 3.6, 31.9%, Hepascore: 0.3, 24.6%, APRI: 1.4, 5.3% of patients (P<0.003, respectively, between tests). By calculating a specific test for cirrhosis, including the FibroMeter markers, the correct classification (93.0%) was significantly higher for the cirrhosis diagnosis compared with the original FibroMeter (90.9%, P=0.005). This specific test provided a 100% positive predictive value for cirrhosis diagnosis versus 88% for original FibroMeter. Using the most accurate original test, cirrhosis can be excluded in 47.5% of patients and is correctly diagnosed, as significant fibrosis, in 100% of patients. A specific test for cirrhosis provides a significant gain in diagnostic accuracy to 93% and in positive predictive value to 100% compared with the original test.

Re-evaluation of the genetic cirrhosis risk score in patients with chronic hepatitis C virus infection

Aims: Recently based on a genome-wide scan, a novel "gene signature" consisting of 7 single nucleotide polymorphisms (SNPs) has been proposed to identify patients at risk for cirrhosis in chronic hepatitis C virus (HCV) infection (Huang et al. Hepatology 2007;46: 297–306). However, this cirrhosis risk score (CRS) has yet to be replicated in an independent sample. Thus, our aim now was to evaluate the gene signature in a large independent cohort of patients with chronic HCV infection.

Usefulness of FibroScan for Detection of Early Compensated Liver Cirrhosis in Chronic Hepatitis B

It is difficult to differentiate early compensated cirrhosis from chronic hepatitis solely by clinical features. The aim of this study was to assess the usefulness of liver stiffness measurement (LSM) for detection of early compensated liver cirrhosis in chronic hepatitis B (CHB). Ninety-one consecutive CHB patients who underwent liver biopsy (LB) and LSM were recruited. All patients did not fulfill the clinical criteria for cirrhosis. The cutoff of LSM for cirrhosis was 10.3 kPa. All patients were divided into either group A (cirrhosis) or group B (CHB) according to LB result. The median LSM values of groups A and B were 11.8 and 7.6 kPa, respectively (P < 0.001). The sensitivity, specificity, positive predictive value, and negative predictive value of LSM in predicting cirrhosis were 0.59, 0.78, 0.68, and 0.72, respectively. The area under the receiver operating characteristics curve (AUROC) of LSM was 0.803, whereas the AUROCs of aspartate to alanine aminotransferase ratio (AAR) and aspartate aminotransferase to platelet ratio index (APRI) were 0.488 and 0.723, respectively. LSM showed an acceptable diagnostic accuracy for detecting early compensated cirrhosis in CHB.

Metabolic syndrome increases the risk of liver cirrhosis in chronic hepatitis B

Background:Metabolic syndrome is associated with non-alcoholic steatohepatitis and cryptogenic cirrhosis. Whether metabolic syndrome affects the severity of chronic hepatitis B (CHB) is unclear.Aim:We aimed to study the relationship between metabolic...

Brief Communication: The Relationship of Regression of Cirrhosis to Outcome in Chronic Hepatitis C

The effect of regression of cirrhosis in chronic hepatitis C is unknown. To evaluate the relation between regression of cirrhosis and clinical outcome in patients with chronic hepatitis C after antiviral therapy. A cohort of patients with cirrhosis treated between 1988 and 2001. Hepatology unit of a tertiary care center in France. 96 patients with chronic hepatitis C and biopsy-proven cirrhosis (METAVIR score F4) who were treated with an interferon-based regimen and had at least 1 posttreatment liver biopsy. Patients were followed until November 2006. Occurrence of a combined end point of liver-related events (ascites, hepatic encephalopathy, variceal bleeding, spontaneous bacterial peritonitis, hepatocellular carcinoma, or liver transplantation) and death in patients with regression of cirrhosis (defined as a decrease from 4 to <or=2 METAVIR units on posttherapy liver biopsy). The median follow-up was 118 months (interquartile range, 86 to 138 months). Eighteen patients had regression of cirrhosis. The incidence of the combined end point per 100 patient-years was 0 in patients with regression of cirrhosis and 4 in patients without regression of cirrhosis (P = 0.002, log-rank test). The transplantation-free survival rate at 10 years was 100% in patients with regression of cirrhosis and 74.2% in patients without regression of cirrhosis (P = 0.025). Selection of patients was retrospective; selection and survival biases may have influenced the estimates of the overall rate of regression of cirrhosis. The low number of patients who experienced regression of cirrhosis precludes analysis of factors that could predict regression of cirrhosis. Regression of cirrhosis occurs after antiviral therapy in some patients with chronic hepatitis C. Regression is associated with decreased disease-related morbidity and improved survival.

Transient elastography (FibroScan)

Transient elastography (FibroScan) is a non-invasive method proposed for the assessment of hepatic fibrosis in patients with chronic liver disease by measuring liver stiffness. It can be easily performed at the bedside or in the outpatients clinic with immediate results and good reproducibility. Limitations include failure (no value) in around 5% of cases, mainly in patients with substantial thoracic fat. In France, FibroScan or FibroTest are recommended for the initial evaluation of liver fibrosis in previously untreated patients with chronic hepatitis C and no concomitant health disorders. FibroScan is validated for the diagnosis of significant fibrosis and cirrhosis in chronic hepatitis C, recurrence of hepatitis C after liver transplantation, co-infections in HIV-HCV patients and chronic cholestatic diseases, but needs further evaluation in other chronic liver diseases. FibroScan is an excellent tool for early detection of cirrhosis and evaluation of portal hypertension, for which it may have prognostic value as well. Studies are needed using FibroScan for the follow-up of patients with and without treatment, and for the screening of patients at risk of liver disease. However, although FibroScan is a good method for the evaluation of fibrosis in patients with chronic liver disease, it has to be borne in mind that FibroScan evaluates liver stiffness. Therefore, FibroScan values have to be interpreted according to clinical, biological and morphological data.

Association of interferon regulatory factor ‐7 gene polymorphism with liver cirrhosis in chronic hepatitis C patients

Interferon (IFN) regulatory factor 7 (IRF-7) has been shown to play an essential role in the transcriptional activation of virus-inducible cellular genes, especially IFN genes. Polymorphisms of the IRF-7 gene may probably affect both the quality and the quantity of IRF-7. We investigated the role of IRF-7 polymorphisms in Japanese patients with chronic hepatitis C virus (HCV) infection. We studied a total of nine polymorphisms of the IRF-7 gene including SNP1047A/G (Lys/Glu) and SNP2157A/G (Gln/Arg) using the Taqman allelic discrimination and sequencing techniques in 406 Japanese patients with chronic HCV infection. We further performed functional analysis of SNP1047 and SNP2157 by transcriptional activation of the IFNA promoter. We found that SNP1047AG and SNP2157AG genotypes were in complete linkage disequilibrium and were present in a significantly higher proportion in HCV-infected patients with cirrhosis (5.6%) than in those without cirrhosis (1.7%) (P=0.03). Multivariate analysis also revealed that SNP1047 and SNP2157 were independently associated with cirrhosis at an odds ratio of 2.5. Functional analysis revealed that SNP1047G and SNP2157G alleles increased IFNA expression. SNP1047AG and SNP2157AG genotypes were strongly associated with cirrhosis. SNP1047G and SNP2157G alleles might be used as markers of host factors associated with a higher risk of cirrhosis in Japanese patients with chronic HCV infection.

Diagnosis and Quantitation of Fibrosis

Hepatic fibrosis is the final common pathway for many different liver insults. Originally considered to be irreversible, hepatic fibrosis is now known to be a dynamic process with a significant potential for resolution. The diagnosis and quantitation of fibrosis have traditionally relied on liver biopsy. However, there are a number of drawbacks including the invasive nature of the procedure, sampling error, and interobserver variability. This article reviews the current role of liver biopsy in the assessment of hepatic fibrosis and discusses the role of the newer noninvasive methods including serum markers and radiologic tests.

Prevalence of Fibrosis and Cirrhosis in Chronic Hepatitis B: Implications for Treatment and Management

To document the prevalence and factors associated with severe fibrosis and cirrhosis in a large population of Asian chronic hepatitis B (CHB) patients. Transient elastography was performed in unselected CHB patients. Liver stiffness score of <8.1 kPa was used as a cut-off for the presence of severe fibrosis or liver cirrhosis. 1315 patients were recruited, of which 951 (72%) were treatment-naive. Of these, 319 (34%) had severe fibrosis, with higher prevalence seen in males compared with females (39% vs 24% respectively, p < 0.01. Severe fibrosis was seen with increasing age from 20% in patients <25 years to 81% in those >65 years. Higher prevalence of severe fibrosis was seen in HBeAg(+) patients compared to HBeAg(-) patients age >45 years (58% vs 43% respectively, p = 0.03), in patients with HBV DNA levels >or=4 log compared with <4 log copies/ml (41% vs 27% respectively, p < 0.01), and in patients with stepwise increase of ALT levels (<0.5 x ULN vs 0.5-1 x ULN vs 1-2 x ULN; 11% vs 30% vs 48% respectively, p < 0.01). After multivariate analysis, gender, age and ALT levels were significant factors associated with severe fibrosis. Patients who received antiviral treatment had lower ALT, stiffness score and prevalence of cirrhosis compared to treatment-naive patients [25 vs 35 U/L (p < 0.01), 6.2 vs 6.7 kPa (p = 0.031) and 14% vs 22% (p = 0.008) respectively]. The overall prevalence of severe fibrosis in CHB patients was 34% with higher rates seen in older age groups, males, and in patients with higher ALT levels.

13C-methacetin-breath test compared to also noninvasive biochemical blood tests in predicting hepatic fibrosis and cirrhosis in chronic hepatitis C

Background The 13C-methacetin-breath test and also several noninvasive blood tests comprising routine laboratory parameters have been proposed to predict fibrosis and cirrhosis in chronic hepatitis C. The aim of the study was to compare the diagnostic accuracy between these tests referring to hepatic histology as gold standard. Methods 96 patients with chronic hepatitis C virus infection underwent percutaneous liver biopsy and the 13C-methacetin-breath test. The Fibroindex, the aspartate aminotransferase to platelet ratio index , and the aspartate aminotransferase to alanine aminotransferase ratio were used as parameters for the staging of fibrosis. The main endpoint was the area under the characteristic curves for the diagnosis of advanced fibrosis (F3-F4) and cirrhosis (F4) according to the Batts Ludwig criteria. Results ROC analysis revealed a cut-off <14.6‰ best with 92.6% sensitivity and 84.1% specificity for the 13C-methacetin-breath test, for the Fibroindex >1.82 70.4% sensitivity and 91.3% specificity, for the aspartate aminotransferase to platelet ratio >1.0 a 66.7% sensitivity and 75.4% specificity, and for the aspartate aminotransferase to alanine aminotransferase ratio >1.0 63.0% sensitivity and 59.4% specificity in predicting liver cirrhosis. The areas under the curve for the breath test, the Fibroindex, aspartate aminotransferase to platelet ratio and the aspartate aminotransferase to alanine aminotransferase ratio were 0.958, 0.845, 0.799, and 0.688, respectively, when predicting cirrhosis. For identifying patients with advanced fibrosis, the areas under the curve were 0.827, 0.804, 0.779, and 0.561, respectively. Discordances between Fibroindex (21%), aspartate aminotransferase to platelet ratio (29%) or aspartate aminotransferase to alanine aminotransferase ratio (37.6%) and liver biopsy were significantly more frequent than between 13C-breath test (11.6%) and liver biopsy ( P < 0.05). Conclusion The 13C-methacetin-breath test is more reliable in predicting advanced fibrosis and cirrhosis than simple biochemical parameters (aspartate aminotransferase to platelet ratio; aspartate aminotransferase to alanine aminotransferase ratio).

Retrospective Evaluation of Serum Markers APRI and AST/ALT for Assessing Liver Fibrosis and Cirrhosis in Chronic Hepatitis B and C Patients with Hepatocellular Carcinoma

Aspartate aminotransferase-to-platelet ratio index (APRI), aspartate aminotransferase-to-alanine aminotransferase (AST/ALT) ratio, platelet count, AST, albumin, bilirubin and alkaline phosphatase were retrospectively evaluated for the prediction of advanced liver fibrosis and cirrhosis in patients with resectable hepatocellular carcinoma in this study. In total, the 97 selected patients consisted of 9 (9.3%) patients with non-B, non-C chronic hepatitis, 48 (49.5%) patients with chronic hepatitis B (CHB) and 40 (41.2%) patients with chronic hepatitis C (CHC). The APRI, but not AST/ALT or other serum markers, showed a significant correlation with advanced liver fibrosis and cirrhosis (p<0.05). The area under receiver operating characteristic curves (AUROC) for predicting advanced fibrosis was 0.69 in CHB patients and 0.87 in CHC patients, whereas AUROC for predicting cirrhosis was 0.75 in CHB patients and 0.84 in CHC patients. In addition, the sensitivity and specificity of APRI were greater than 80% for predicting advanced fibrosis and cirrhosis in the CHC patients. APRI is a simple and non-invasive biochemical marker of liver fibrosis and cirrhosis, particularly in CHC patients. APRI potentially could be used to decrease the number of liver biopsies.

Susceptibility to active decompensated cirrhosis is associated with polymorphisms of intercellular adhesion molecule‐1 (ICAM‐1) in chronic HBV carriers

Intercellular adhesion molecule-1 (ICAM-1) plays an important role in the pathogenesis of viral hepatitis B. Several inflammatory diseases are associated with distinct polymorphisms of the ICAM-1 gene. The aims of this study were to analyse the association of ICAM-1 polymorphisms G241R and K469E with susceptibility to active decompensated cirrhosis in chronic hepatitis B virus (HBV) carriers. The polymorphisms at codons G241R and K469E of ICAM-1 were analysed by sequence-specific primer polymerase chain reaction (PCR-SSP) in 572 unrelated chronic HBV carriers and 157 unrelated healthy HBV non-infected blood donors. There were significantly increased frequencies of R at codon 241 and E at codon 469 in patients with active decompensated cirrhosis (38.3% and 58.3%), compared with patients with chronic hepatitis B (CHB; 21.9% and 46.5%) and chronic asymptomatic HBV carriers (AsC; 12.6% and 40.3%). The frequencies of R241-E469 haplotype and genotypes carrying at least one R241-E469 haplotype were significantly higher in patients with active decompensated cirrhosis than those in patients with CHB (38.3% and 63.3%vs 21.9% and 36.7%), and significantly higher in patients with CHB than those in AsC (21.9% and 36.7%vs 12.6% and 23.3%). The ICAM-1 polymorphisms at codons G241R and E469K were associated with the disease susceptibility, and susceptibility to active decompensated cirrhosis is significantly increased in chronic HBV carriers carrying at least one R241-E469 haplotype.

Tenascin-C in Chronic Canine Hepatitis: Immunohistochemical Localization and Correlation with Necro-Inflammatory Activity, Fibrotic Stage, and Expression of Alpha-Smooth Muscle Actin, Cytokeratin 7, and CD3+ Cells

During fibrosis, the extracellular matrix (ECM) is continuously remodeled and increases in volume due to the production of various proteins. We studied the distribution of tenascin-C (TN-C) and the correlation of TN-C with the necro-inflammatory activity and expression of alpha-smooth muscle actin (alpha-SMA), cytokeratin 7 (CK7), and CD3+ T-lymphocytes in canine chronic hepatitis. This was analyzed using immunohistochemistry and semiquantitative scoring. We used 3 groups (n = 19) of dogs: group 1 (n = 5) with neonatal hepatitis/lobular dissecting hepatitis (NH/LDH), group 2 (n = 8) with chronic hepatitis/cirrhosis (CH/CIRR), and group 3 (n = 6) consisting of healthy animals. In normal livers, TN-C was localized in Disse's space and around bile ducts and blood vessels. In CH/CIRR livers, TN-C was localized at the periphery of the regenerating nodules and was conspicuous in the bridging fibrous bands. In NH/LDH, TN-C was diffusely distributed along the reticular fibers that dissected between single cells or groups of hepatocytes. alpha-SMA in the normal hepatic parenchyma showed an irregular distribution along the perisinusoidal linings. In other groups, alpha-SMA was increased in fibrotic septa and perisinusoidal linings. In normal livers, CK7 was positive in bile ducts. In other groups, CK7-expressing cells were conspicuous in the portal-parenchymal interface, the periphery of the regenerative nodules, and the degenerated parenchyma. The pattern of CD3+ lymphocytes was inversely proportional to that of TN-C. These results also showed that TN-C is strongly correlated with increased fibrotic stage, inflammatory activity, and expression of CK7 and alpha-SMA. TN-C, CK7, and CD3 expression did not differ between diagnostic groups.

Pre-S Deletion and Complex Mutations of Hepatitis B Virus Related to Advanced Liver Disease in HBeAg-Negative Patients

This longitudinal study investigated the interactions and roles of hepatitis B virus (HBV) genotypes, pre-S deletions, and core promoter and precore mutations on the progression of liver disease in hepatitis B e antigen (HBeAg)-negative patients. A total of 141 HBeAg-negative patients without liver cirrhosis or hepatocellular carcinoma at study entry were recruited for this study, including 45 inactive HBV carriers and 96 patients with HBeAg-negative chronic hepatitis B. The HBV genotypes and the sequences of pre-S, core promoter, and precore regions were determined. Compared with patients without developing liver cirrhosis, patients with the development of liver cirrhosis had higher rates of genotype C; pre-S deletions; C or G1753, T1762/A1764, T1766, and/or A1768 mutants; and G1799 variant. Cox regression analysis showed that older age, higher total bilirubin and HBV DNA levels, pre-S deletions, and T1766 and/or A1768 mutants were significantly associated with the development of liver cirrhosis. HBV with a complex mutation pattern (pre-S deletion, T1762/A1764, and T1766 and/or A1768 mutants) rather than a single mutation was associated with the development of liver cirrhosis, and the patterns of mutation combinations differed between HBV genotype B and C. Moreover, pre-S deletion was a significant risk factor for hepatocellular carcinoma. This study indicated that pre-S deletion and combined mutations of HBV are useful molecular markers for predicting the clinical outcomes of HBeAg-negative patients.

Diagnostic accuracy of the 13C-methacetin breath test and noninvasive biochemical blood tests in predicting hepatic fibrosis and cirrhosis in chronic hepatitis C

Aims: The 13C-methacetin breath test and several also noninvasive blood tests comprising routine laboratory parameters have been proposed to predict fibrosis and cirrhosis in chronic hepatitis C.

Sustained virological response to interferon reduces cirrhosis in chronic hepatitis C: a 1386‐patient study from Taiwan

The long-term benefits of interferon-based therapy on preventing cirrhosis at non-cirrhotic stage in chronic hepatitis C patients are not fully clarified. To evaluate the effectiveness of interferon-based therapy regarding to cirrhosis prevention in non-cirrhotic chronic hepatitis C patients. A total of 1386 biopsy-proven, non-cirrhotic chronic hepatitis C patients (892 received interferon-based therapy and 494 untreated) were enrolled. Fifty-six untreated and 51 treated (24 sustained virologic responders and 27 non-responders) patients developed cirrhosis during a mean follow-up period of 5.0 (1-16) and 5.1 (1-15.3) years, respectively. The annual incidences of cirrhosis in untreated and treated groups were 2.26 and 1.11% (non-responders: 1.99%, sustained responders: 0.74%), respectively. The 15-year cumulative incidence of cirrhosis was significantly lower in treated (9.9%) than untreated patients (39.8%, P = 0.0008, log-rank test). The 14.5-year cumulative incidence of cirrhosis was significantly lower in sustained responders (4.8%) compared with non-responders (21.6%, P = 0.0007) and untreated patients (36.6%, P < 0.0001). The difference was not significant between non-responders and untreated controls. Cox proportional hazards regression showed sustained virologic responders and younger age were independent negative factors for cirrhosis development. A sustained virologic response secondary to IFN-based therapy could reduce cirrhosis development in chronic hepatitis C patients.

Noninvasive diagnosis of hepatic fibrosis in chronic hepatitis C

Assessment of hepatic fibrosis is important for determining prognosis, guiding management decisions, and monitoring disease. Histological evaluation of liver biopsy specimens is currently considered the reference test for staging hepatic fibrosis. Since liver biopsy carries a small but significant risk, noninvasive tests to assess hepatic fibrosis are desirable. This editorial gives an overview on noninvasive methods currently available to determine hepatic fibrosis and their diagnostic accuracy for predicting significant fibrosis and cirrhosis in chronic hepatitis C. Based on available data, the performance of simple tests derived from routine laboratory parameters appears to be similar to that of more complex and expensive fibrosis panels. Transient elastography seems more accurate than blood tests for diagnosing cirrhosis.

Comparison of simple tests for the non‐invasive diagnosis of clinically silent cirrhosis in chronic hepatitis C

Biopsy is the gold standard for assessing cirrhosis in patients with chronic hepatitis C virus infection, but it is expensive and at risk of complications. Alternative non-invasive methods have been developed but their usefulness remains uncertain. To compare the accuracy of five non-invasive scores in detecting cirrhosis. We reviewed the charts and liver biopsies of 228 consecutive, treatment-naïve, hepatitis C virus-positive patients, 13.2% of whom with histological diagnosis of cirrhosis. The five alternative scores were age-platelet index, cirrhosis discriminant score, aspartate transaminases to platelet ratio index, Pohl's index, and aspartate transaminases/alanine transaminases ratio. The specificities of the scores were good (87-100%), but not so their sensitivities (17-67%). Accordingly positive likelihood ratios were generally good but negative likelihood ratios were suboptimal. Combinations of the scores independently related to cirrhosis only slightly change this diagnostic accuracy. Using double cut-offs to exclude/diagnoses cirrhosis, cirrhosis discriminant score classified 21% of patients without misdiagnoses and aspartate transaminases to platelet ratio index classified 85% of case with 9% of misdiagnoses. The five scores showed variable sensitivities and specificities in detecting liver cirrhosis, both individually and in combination. The use of double cut-off points may make the cirrhosis discriminant score and aspartate transaminases to platelet ratio index useful to reduce the number of patients submitted to liver biopsy.