Disclosures: Kazuaki Chayama – Consulting: Abbvie;

Grant/Research Support: Dainippon Sumitomo, Chugai, Mitsubishi Īanabe, DAIICHI SANKYO, Toray, BMS, MSD; Speaking and Teaching: Chugai, Mitsubishi Īanabe, DAIIcHl SANKYO, KYORIN, Nihon Medi-Physics, BMS, Dainippon Sumitomo, MSD, ASKA, Astellas, AstraZeneca, Eisai, Olympus, GlaxoSmithKline, ZERIA, Bayer, Minophagen, Shinyaku, Takeda, AJINOMOTO, Meiji Seika, Toray The following people have nothing to disclose: Tsunehiro Ochi, Motoi Hashiba, Masafumi Ono, Hideyuki Hyogo, Yukio Ikeda, Kensuke Munekage, Nobuto Okamoto, Shinji Iwasaki, Yuichiro Eguchi, Toshiji Saibara Background/Aims: Gallstone disease and fatty liver are both prevalent diseases in the general populations and share the same risk factors Galunisertib such as obesity and insulin resistance. However, association between gallstone disease and ultrasonographically diagnosed fatty liver has not been completely established. The aim of this study was to characterize the relationship between gallstone

disease and fatty liver in large population. Methods: A cross-sectional study with 24, 050 health check-up subjects was conducted. Gallstone disease was defined as the presence of gallstones on abdominal sonography or previous history of cholecystectomy. Fatty liver was diagnosed on the basis of typical ultrasonographic findings. Subjects positive for hepatitis B or C virus or PLX-4720 cell line with a history of other forms of hepatitis were excluded. Results: The mean age of the subjects was 48.7 ± 11.1 years and 54.5% were male. The prevalence of gallstone disease was 5.3% (n=1, 280). The prevalence of fatty liver increased with presence of gallstone disease (43.0% vs.31.3%, p <0.001). In the same manner, the prevalence of gallstone disease increased with presence of fatty liver (7.2% vs.4.4%, p <0.001). The gallstone disease was significantly associated with fatty liver after adjusted for age 上海皓元 and sex [odds ratio (OR) 1.50 95%

confidence interval (Cl) 1.331.69]. Multivariate regression analysis after adjustment for body mass index, waist circumference, total cholesterol, triglycerides, HDL cholesterol, HbA1 c, and systolic blood pressure showed that gallstone disease was statistically significantly associated with fatty liver (OR 1.23, 95% CI 1.06-1.42, p=0.007). These association was attenuated, however still statistically significant after adjusting for insulin resistance (OR 1.27 95% Cl 1.04-1.55, p=0.018). Conclusions: Patients with fatty liver have a high prevalence of gallstone disease. Gallstone disease is associated with fatty liver independently of known metabolic risk factors, especially insulin resistance.

Eleven groups received diet-only interventions, two exercise-only, and 19 diet and physical activity/exercise.

Studies consistently showed reductions in liver fat and/or serum aminotransferase concentration, with the strongest correlation being with weight reduction. Of the five studies reporting changes in hepatic histopathology, all showed a trend toward reduction in inflammation; in two, this was statistically significant. this website Changes in liver fibrosis were less consistent, with only one study showing a significant reduction. The majority of studies also reported improvements in glucose control/insulin sensitivity following intervention. In addition, lifestyle modifications leading to weight loss diet and/or increased physical activity consistently selleck chemicals improved glucose control/insulin sensitivity. Another systematic review and meta-analysis of randomized trials by Musso et al.

suggests that lifestyle-induced weight loss is safe and improved cardiometabolic risk profile; a weight loss ≥ 7% improved hepatic histological disease activity but was achieved by < 50% patients.[29] A weight loss of 5% is considered clinically important by the US Food and Drug Administration.[10] The results of several original articles published in the past 2 years also show that diet-induced weight loss reduces liver enzymes and hepatic steatosis (Table 3).[30-32] Therefore, the US Practice Guideline for the Diagnosis and Management of NAFLD recommend that weight loss achieved either by hypocaloric diet alone or in conjunction with increased physical activity generally reduces hepatic steatosis. At least 3–5% of weight loss appears necessary to improve steatosis, but a greater weight loss (∼ 10%) may be needed to improve necroinflammation.[1] Observational n = 16 (obese adults) non-controlled clinical intervention n = 59 (obese women) Observational study n = 71 (obese children, partly with NAFLD) RCT n = 61 (NAFLD) RCT n = 60 (children with biopsy-proven NAFLD) Observational MCE公司 study n = 11 (obese women without diabetes) RCT n = 116 (8–17 years, biopsy-proven

NAFLD) RCT n = 28 (biopsy-proven NAFLD) RCT n = 66 (biopsy-proven NASH) Diets to promote weight loss or to maintain a stable body weight are generally divided into four categories: low fat, low carbohydrate, low calorie, and very low calorie (Table 4).[8-10] A meta-analysis of six randomized, controlled trials (RCTs) found no difference between the main diet types (low calorie, low carbohydrate, and low fat), with a 2–4 kg (approximately 4% of baseline weight) weight loss at 12–18 months in all studies.[9] The very low-calorie diets are not recommended for general use, as they are associated with adverse side-effects and only prescribed on a case-to-case basis for rapid weight loss (about 1.5–2.5 kg/week) in some severe obese patients.[8, 9] However, large fluctuations in weight can exacerbate liver injury and result in liver failure in patients with NASH.

Then the cells were collected for messenger RNA (mRNA) quantification and the supernatants were collected for IL-17A detection. Total RNA was extracted Ivacaftor order from sorted CD4+ T cells and HBcAg-stimulated cells using the RNeasy Mini Kit (Qiagen, Santa Clarita, CA) according

to the manufacturer’s instructions. The RNA was reverse-transcribed to complementary DNA (cDNA) using oligo (dT) primers at 42°C for 30 minutes and at 95°C for 5 minutes. Quantitative expressions of the RORγt and IL-17A transcripts were determined by staining with the fluorogenic dye SYBR Green using the reported primers and methods.15 GAPDH was used to normalize the samples in each PCR reaction.12 The absence of nonspecific primer-dimer products was verified by melting-curve and gel-migration analyses. Results are expressed in terms of relative mRNA quantification calculated by using the arithmetic formula 2−ΔCt. A cytometric bead assay (Bender selleck kinase inhibitor Medsystems, Copenhagen, Denmark) was employed to measure levels of IL-17, IL-23 p19, IL-1β, IL-6, IL-12 p35, interferon (IFN)-γ, IL-22, IL-8, and GRO-α of plasma and supernatants according to described protocols.30 Paraffin-embedded, formalin-fixed liver tissue (5 μm) was incubated with

anti-IL-17 (AF-317-NA, R&D Systems, Minneapolis, MN) antibody overnight at 4°C after blocking endogenous peroxidase activity with 0.3% H2O2. 3-Amino-9-ethyl-carbazole (red color) was used as the substrate followed by counterstaining with hematoxylin for single staining. Double staining was performed by using the avidin-biotin-peroxidase system with two different substrates: vector blue (blue color) for IL-17, and 3-amino-9-ethyl-carbazole for CD4. Positively stained cells were counted at high-power field (hpf, ×400) according to described protocols.10–12 The virological assay was performed according to our described protocols.10–12 The limit of detection of the assay was 500 copies/mL. All data were analyzed using SPSS software (Chicago, IL) and are summarized as means and standard

deviations. 上海皓元医药股份有限公司 Comparison between various individuals was performed using the Mann-Whitney U test. Comparison between the same individual was performed using the Wilcoxon matched-pairs T test. Correlation analysis was evaluated by the Spearman rank correlation test. For all tests, two-sided P < 0.05 was considered statistically significant. We first identified peripheral IL-17–producing cells in vitro by way of PMA/ionomycin stimulation. IL-17–producing cells were mainly comprised of CD4+ T cells; in contrast, CD8+ T cells, monocytes, natural killer (NK) cells, B cells, mDCs, and γδ T cells expressed low levels of IL-17 (Fig. 1A). Phenotypic analysis indicated that IL-17+CD4+ T cells expressed high levels of the memory marker CD45RO, but low levels of CD45RA, CD57 (a senescence marker), and Ki67 (a proliferation marker) (Fig. 1B).

Then the cells were collected for messenger RNA (mRNA) quantification and the supernatants were collected for IL-17A detection. Total RNA was extracted Epigenetics Compound Library from sorted CD4+ T cells and HBcAg-stimulated cells using the RNeasy Mini Kit (Qiagen, Santa Clarita, CA) according

to the manufacturer’s instructions. The RNA was reverse-transcribed to complementary DNA (cDNA) using oligo (dT) primers at 42°C for 30 minutes and at 95°C for 5 minutes. Quantitative expressions of the RORγt and IL-17A transcripts were determined by staining with the fluorogenic dye SYBR Green using the reported primers and methods.15 GAPDH was used to normalize the samples in each PCR reaction.12 The absence of nonspecific primer-dimer products was verified by melting-curve and gel-migration analyses. Results are expressed in terms of relative mRNA quantification calculated by using the arithmetic formula 2−ΔCt. A cytometric bead assay (Bender AZD1208 solubility dmso Medsystems, Copenhagen, Denmark) was employed to measure levels of IL-17, IL-23 p19, IL-1β, IL-6, IL-12 p35, interferon (IFN)-γ, IL-22, IL-8, and GRO-α of plasma and supernatants according to described protocols.30 Paraffin-embedded, formalin-fixed liver tissue (5 μm) was incubated with

anti-IL-17 (AF-317-NA, R&D Systems, Minneapolis, MN) antibody overnight at 4°C after blocking endogenous peroxidase activity with 0.3% H2O2. 3-Amino-9-ethyl-carbazole (red color) was used as the substrate followed by counterstaining with hematoxylin for single staining. Double staining was performed by using the avidin-biotin-peroxidase system with two different substrates: vector blue (blue color) for IL-17, and 3-amino-9-ethyl-carbazole for CD4. Positively stained cells were counted at high-power field (hpf, ×400) according to described protocols.10–12 The virological assay was performed according to our described protocols.10–12 The limit of detection of the assay was 500 copies/mL. All data were analyzed using SPSS software (Chicago, IL) and are summarized as means and standard

deviations. MCE Comparison between various individuals was performed using the Mann-Whitney U test. Comparison between the same individual was performed using the Wilcoxon matched-pairs T test. Correlation analysis was evaluated by the Spearman rank correlation test. For all tests, two-sided P < 0.05 was considered statistically significant. We first identified peripheral IL-17–producing cells in vitro by way of PMA/ionomycin stimulation. IL-17–producing cells were mainly comprised of CD4+ T cells; in contrast, CD8+ T cells, monocytes, natural killer (NK) cells, B cells, mDCs, and γδ T cells expressed low levels of IL-17 (Fig. 1A). Phenotypic analysis indicated that IL-17+CD4+ T cells expressed high levels of the memory marker CD45RO, but low levels of CD45RA, CD57 (a senescence marker), and Ki67 (a proliferation marker) (Fig. 1B).

Luc in Montréal, Quebec) and a community-based hepatology clinic (the Liver Centre in Toronto, Ontario) between July 2009 and July 2010. Patients meeting any of the following criteria were ineligible: (1) contraindications to LSM (e.g., pregnancy, ascites, implantable cardiac devices, etc.); (2) BMI <28 kg/m2; (3) previous liver transplant; (4) known malignancy or other terminal disease;

and (5) refusal to undergo a liver biopsy. Health Canada and the research ethics boards of the participating institutions approved the study protocol (clinicaltrials.gov, NCT 00926224). The study sponsor (Echosens; PLX-4720 mw Paris, France) oversaw data collection and monitoring, but had no role in data analysis, drafting of the article, or in the decision to submit the article for publication. Before TE examination, demographic details, etiology of liver disease, and anthropometric measurements (weight, height, BMI, waist circumference, and thoracic perimeter measured at the xiphoid process) were obtained. Biochemical data including liver biochemistry, platelets, albumin, and fasting glucose, cholesterol, and triglycerides from within 6 months of screening

were recorded. Presence of the metabolic syndrome was defined according to guidelines of the American Heart Association and National Heart, Lung, and Blood Institute.17 Nine experienced operators at the five study sites performed all FibroScan examinations as per the manufacturer’s recommendations. All operators had completed at least 50 prior exams (four had performed >500 exams; one had >200; three had >100; and one selleck inhibitor had >50). Briefly, with the patient lying in the dorsal decubitus position the tip of the transducer probe was placed on the skin between the ribs over the right lobe of the liver. Assisted by a sonographic image, a portion of the liver at least 6 cm thick and free of large vascular structures was MCE公司 identified using a portable 10 MHz ultrasound transducer (Mindray DP-6600; Mindray, Shenzhen, China).

At this site the distance between the skin and liver capsule (skin-capsular distance) was measured and an attempt was made to collect at least 10 valid measurements with each of the M and XL probes. Specific differences between the M and XL probes include their central ultrasound frequency (3.5 versus 2.5 MHz), vibration amplitude (2 versus 3 mm), diameter of their tips (9 versus 12 mm), and measurement depth from the skin surface (25-65 versus 35-75 mm).15 The manufacturer recommends that the XL probe be used in patients with a skin-capsular distance ≥25 mm. Examinations with no successful measurements after at least 10 attempts were deemed failures. The median liver stiffness value (in kPa) was considered representative of the elastic modulus of the liver. As an indicator of variability, the ratio of the interquartile range (IQR) of liver stiffness to the median value (IQR/M) was calculated.

Based on retrospective analyses of the IDEAL trial,11 the sponsor proposed that an HCV-RNA decline ≤1.0 log10 at week 4 was predictive of the more traditionally defined null response on P/R (i.e., <2 log10 HCV RNA decline at week 12). SVR rates for SPRINT-II subjects administered BOC who had <1.0 log10 decline at week 4 were 28% in Arm 2 and 38% in Arm 3, compared to 4% in the P/R control arm. The FDA's SVR analysis selleckchem of treatment-naïve subjects in SPRINT-II demonstrated that 31% (26/83) of subjects with <1.0 log10 decline at week 4 in the P/R control

arm (Arm 1) would be incorrectly classified as null responders10 (the remaining subjects discontinued treatment, had a partial response, or relapsed). To obtain a more conservative estimate of the SVR rate in null responders, an alternative surrogate definition of <0.5 log10 HCV RNA decline at week 4 was investigated. Based on the end of study outcomes (i.e., null responder, partial responder, relapser,

or responder achieving SVR) for such subjects in SPRINT-II treated with P/R (n = 25), 22 subjects were null responders (88%), one was a partial responder, and two discontinued treatment. The SPRINT-II study design allowed us to analyze outcomes of treatment-naïve subjects who were treated with BOC (Arms 2 and 3) and had similar interferon responsiveness during the lead-in period. The SVR rates in these poorly interferon responsive subjects (defined here as <0.5 log10 decline in HCV RNA at week 4) who received BOC Selleckchem Dabrafenib treatment was 28% (Arm 2: n/N = 13/47) and 30% (Arm 3: n/N = 11/37). By comparison, the observed SVR rate for poorly interferon responsive subjects treated with P/R (i.e., those with ≤0.5 log10 HCV RNA decline at week 4) was 0%. Whether a medchemexpress <1.0

or <0.5 log10 decline in HCV RNA at week 4 was used to categorize poorly interferon responsive subjects from SPRINT-II, a treatment benefit was demonstrated in the BOC regimens over treatment with P/R alone. Further, it is important to consider the reasons behind equating prior null responders and treatment-naïve subjects with poor interferon responsiveness. The previous analysis demonstrated that BOC provided meaningful benefit in treatment-naïve subjects with interferon response characteristics similar to prior null responders. However, to consider using data from treatment-naïve patients to predict response in P/R-experienced patients, one needs to demonstrate that P/R response remains similar after a second course of P/R treatment. To address this question we assessed the relationship between virologic responsiveness through week 4 and treatment outcome for P/R arm subjects in SPRINT-II (i.e., log10 decline in HCV RNA at week 4 grouped according to end of study outcome) (Fig. 2). Similarly, subjects from RESPOND-II were grouped according to previous treatment outcome (relapser or partial responders) (Fig. 2).

In order to inhibit mature hepatocytes regeneration and increase selleck hepatic progenitor cell expansion and differentiation, the treated rats were fed with 2-AAF during the period of cirrhosis progression. The results showed that the expressions of hepatic oval cell markers (OV6 and CK19) were increased significantly during fibrosis progression. In the CCl4/ 2-AAF treated rats, OV6 positive cells

and CK19 positive cells extended across the liver lobule, formed bridges between portal tracts and divided the parenchyma into smaller pseudolobules, as determined by immunohistochemitry. Double staining showed that OV6 was largely colocated with α-SMA positive cells, and the number of cells positive for both OV6 and α-SMA was obviously increased after administration of 2-AAF. The expressions of Wnt4, Wnt5b, frizzled2, frizzled3 and frizzled6 were markedly increased after administration of 2-AAF (p<0.01). Immunohistochemistry showed that p-catenin protein was

mostly localized to the nucleus of cells before administration of 2-AAF; however, p-catenin was found predominantly within the cytoplasm after administration of 2-AAF. In addition, the expression level of p catenin was not changed by the administration of 2-AAF, suggesting that the activation of Wnt pathways was not mediated through the classical p-catenin pathway. Moreover, after administration of 2-AAF, gene expression of frizzled1 and frizzled4 was markedly decreased (p<0.01); however frizzled5 expression was not significantly changed, indicating that non-canonical Wnt signaling rather DAPT order than Wnt/p-catenin signaling was primarily activated. We also determined that the expression of TGF-β1 was markedly increased in vivo after administration of 2-AAF. Expression of α-SMAand F-actin, as well as collagen types MCE I and IV were significantly increased after the WB-F344 cell line, was treated with TGB-p1 for 24 hours. Additional investigation revealed that both Wnt5b and frizzled2 expression were significantly

increased in WB-F344 cells after treatment with TGF-β1 (p < 0.01), and p-catenin expression was not up-regulated during the treatment. Thus, these in vitro results confirmed our finding in vivo. In conclusion, our results indicate that hepatic progenitor cells appear to transdifferentiate into myofibroblasts and exhibit a profibrotic effect in the fibrogenic process through activating the non-canonical Wnt signaling pathway. Disclosures: The following people have nothing to disclose: Jiamei Chen, Yongping Mu, Yuyou Duan, Ping Liu Background: Activation of the FXR and TGR5 bile acid receptor pathways with the dual agonist INT-767 has been shown to improve non-alcoholic fatty liver disease (NAFLD) in a murine diet-induced obesity model. While the mechanisms of the liver improvement remain to be fully elucidated direct effects of these pathways on hepatocyte and macrophage function have been demonstrated.

As noted in the article by Davenport et al.,[1] in addition to “BASM,” another term for infants with BA and stereotypical syndromic abdominal and vascular anomalies is “biliary atresia laterality sequence.” Given that only 70% of our patients with laterality defects actually had splenic anomalies, the latter term might be preferable in the future to MK-2206 clinical trial “BASM” to describe this stereotypical group of infants. The Canadian Pediatric Hepatology Research group has recently reported their analysis of 382 infants with BA and the associated anomalies.[22] Forty-four (13%) had

associated anomalies, only 25 (6.5%) of which were associated with SM. The authors concluded that BA infants with anomalies demonstrated a spectrum of laterality defects and suggested that the meaning of the acronym BASM be modified to “biliary atresia structural malformation.” Our conclusions are somewhat similar in that a total of 16% of our infants were in the anomaly Groups 2 and 3. On the other hand, the main difference between our observations and those of the Canadian group was that Group 2 infants frequently exhibited major birth defects Protein Tyrosine Kinase inhibitor of the genitourinary and/or gastrointestinal systems, not considered part of defective lateralization, suggesting that this group may represent a different etiopathogenesis than Groups 1 and 3. Group 3

infants were younger at the time of initial evaluation compared to Group 1. The associated anomalies in Group 3, especially the cardiac lesions associated with murmurs or cyanosis, probably brought the patient to medical attention sooner than the infants with isolated cholestasis. An unexpected finding was the high incidence of

autoimmunity in first-degree relatives of all BA groups (average 44%). The occurrence of autoimmune diseases in relatives provides circumstantial evidence that a candidate disease (i.e., BA) may be autoimmune in nature.[23] The incidence of autoimmunity in first-degree relatives is much higher than that found in the general population, where autoimmunity rates vary from 2.5%-9%.[26, 27] Importantly, the incidence of autoimmunity G protein-coupled receptor kinase in first-degree relatives of BA patients was similar to the rate of 37%-43% identified in autoimmune hepatitis[26] and 25.5% in type-1 diabetes mellitus.[25] This intriguing finding of autoimmunity in first-degree relatives of BA patients warrants further investigation. The fact that there was no difference in autoimmunity rates between the three groups suggests that the autoimmune hypothesis of BA may be relevant to the pathogenesis of all types of BA and is a clue to be pursued in further studies. It is also possibly that the high incidence simply resulted from our rigorous questionnaire containing a long list of autoimmune diseases and not being of pathogenetic significance.

As noted in the article by Davenport et al.,[1] in addition to “BASM,” another term for infants with BA and stereotypical syndromic abdominal and vascular anomalies is “biliary atresia laterality sequence.” Given that only 70% of our patients with laterality defects actually had splenic anomalies, the latter term might be preferable in the future to find more “BASM” to describe this stereotypical group of infants. The Canadian Pediatric Hepatology Research group has recently reported their analysis of 382 infants with BA and the associated anomalies.[22] Forty-four (13%) had

associated anomalies, only 25 (6.5%) of which were associated with SM. The authors concluded that BA infants with anomalies demonstrated a spectrum of laterality defects and suggested that the meaning of the acronym BASM be modified to “biliary atresia structural malformation.” Our conclusions are somewhat similar in that a total of 16% of our infants were in the anomaly Groups 2 and 3. On the other hand, the main difference between our observations and those of the Canadian group was that Group 2 infants frequently exhibited major birth defects Selleck Ceritinib of the genitourinary and/or gastrointestinal systems, not considered part of defective lateralization, suggesting that this group may represent a different etiopathogenesis than Groups 1 and 3. Group 3

infants were younger at the time of initial evaluation compared to Group 1. The associated anomalies in Group 3, especially the cardiac lesions associated with murmurs or cyanosis, probably brought the patient to medical attention sooner than the infants with isolated cholestasis. An unexpected finding was the high incidence of

autoimmunity in first-degree relatives of all BA groups (average 44%). The occurrence of autoimmune diseases in relatives provides circumstantial evidence that a candidate disease (i.e., BA) may be autoimmune in nature.[23] The incidence of autoimmunity in first-degree relatives is much higher than that found in the general population, where autoimmunity rates vary from 2.5%-9%.[26, 27] Importantly, the incidence of autoimmunity 3-oxoacyl-(acyl-carrier-protein) reductase in first-degree relatives of BA patients was similar to the rate of 37%-43% identified in autoimmune hepatitis[26] and 25.5% in type-1 diabetes mellitus.[25] This intriguing finding of autoimmunity in first-degree relatives of BA patients warrants further investigation. The fact that there was no difference in autoimmunity rates between the three groups suggests that the autoimmune hypothesis of BA may be relevant to the pathogenesis of all types of BA and is a clue to be pursued in further studies. It is also possibly that the high incidence simply resulted from our rigorous questionnaire containing a long list of autoimmune diseases and not being of pathogenetic significance.

Liver sections from 3- and 6-month-old Mdr2−/− and dKO mice displayed progressive periductular inflammation and a broad rim of periductular extracellular matrix, which was detected neither in control nor in Rage−/− animals (Supporting Fig. 2A). Staining of tissue sections from 3- and 6-month-old control, Mdr2−/−, and dKO livers for the panleukocyte marker CD45, the neutrophil marker myeloperoxidase (Fig. 2A,B), and the T-cell marker CD3 (data not shown) revealed highly increased levels of immune cells in

livers of both Mdr2−/− and dKO mice as compared to controls. However, no significant difference was found between Mdr2−/− and dKO liver sections, suggesting that RAGE deficiency had no major impact on the recruitment of inflammatory cells to the liver in the Mdr2−/− mouse. On the contrary, premalignant WT and Rage−/− mice 6 months MK0683 datasheet after DEN injection did not show any evident sign of liver inflammation Antiinfection Compound Library as measured by H&E and immunohistochemical staining for either CD45, myeloperoxidase, or CD3-positive cells (Supporting Fig. 3A and data not shown). At 3 months of age liver and liver/body weight measurements revealed a slight increase in Mdr2−/− and dKO mice as compared to controls, which became significant after 6 months. However, no significant reduction was found in the liver weight of dKO compared to Mdr2−/− mice (Supporting Fig. 2B). Finally, qPCR analysis of tumor necrosis factor alpha (TNF-β), interleukin

(IL)−1, IL-6, and several other cyto- Fossariinae and chemokines revealed comparable transcript levels between Mdr2−/− and dKO livers (Fig. 2C; Supporting Fig. 4). In summary, our results demonstrated that RAGE expression is dispensable for the onset and maintenance of inflammation in the Mdr2−/− model. At 3 months of age, both Mdr2−/− and dKO mice exhibited an increased compensatory proliferation of hepatocytes as compared to controls, while the amount of proliferating cell nuclear antigen (PCNA)-positive hepatocytes was significantly

reduced in 6-month-old dKO mice as compared to matched Mdr2−/− mice (Supporting Fig. 5A). However, we did not observe any difference in Caspase-3 activation in control, Mdr2−/−, and dKO mice (Supporting Fig. 5B). Quantification of serum samples of 3- and 6-month-old mice showed significantly higher ALT levels in Mdr2−/− and dKO mice as compared to controls. However, this increase in ALT levels was more pronounced in Mdr2−/− animals as compared to dKO mice (Fig. 3A). Similar results were observed for aspartate transaminase (AST) levels (data not shown). Fibrosis analysis by Sirius Red histochemistry of Mdr2−/− liver sections revealed strong periportal and septal fibrosis both at 3 and 6 months of age. Interestingly, dKO livers displayed only a mild fibrosis at 3 months that was slightly increased at 6 months of age (Fig. 3B). Impaired fibrosis in dKO livers was further confirmed by qPCR analysis for Collagen1β1 expression (Fig. 3C).