Co,Ltd, Daiichi Sankyo Pharm Co,Ltd, Takeda Pharm Co,Ltd,

Co.,Ltd., Daiichi Sankyo Pharm. Co.,Ltd., Takeda Pharm. Co.,Ltd., AstraZeneca K.K.:,

Eisai Co.,Pharm.Ltd, FUJIFILM Medical Co.,Ltd. The following people have nothing to disclose: Taichiro Nishikawa, Kanji Yama-guchi, Michihisa Moriguchi, Yoshio Sumida, Hironori Mitsuyoshi, Shinji Tanaka, Shigeki Arii Current treatments options MG-132 purchase for HCC are of limited efficacy. Our focus is the development of effective chemoprevention. Accomplishing this will require an understanding of the molecular pathogenesis of HCC. Our work focuses on the mechanistic Target Of Rapamycin (mTOR), a nutrient-sensing serine/ threonine protein kinase that regulates cell cycle progression, protein synthesis, gene expression, and ribosomal biogenesis. Preliminary studies in our lab, using a well-characterized rat model of progenitor-derived HCC, have shown that mTOR is activated in the early stages of preneoplastic foci development. We showed that rapamycin, a specific mTOR inhibitor, blocks this crucial stage of development. This is a pivotal finding that warrants in-depth characterization of the genetic signature and molecular pathogenesis of the rapamycin-inhibited foci as compared to placebo-control, progressive

preneoplastic www.selleckchem.com/products/SB-203580.html foci. Based on this observation of mTOR activation early in preneo-plastic foci development, we hypothesized that inhibition of mTOR signaling learn more during the early window of activation alters the genetic signature of preneoplastic foci. To test this hypothesis, we isolated tissue from foci of rats that have undergone the Solt-Farber protocol to induce HCC. In this protocol, rats are injected with a single dose of the carcinogen

diethylnitrosamine (DENA). Seven days later, they are implanted with a time-release 2-acetylaminofluorene pellet and subjected to 2/3 partial hepatectomy. For the present study, rats were also implanted with a 21-day time-release placebo or rapamycin pellet at time of partial hepatectomy. Liver tissues were harvested 70 days after DENA administration, resulting in a 42-day hiatus between the end of rapamycin administration and tissue harvest. Persistent foci, which were reduced by approximately 80% in the rapamycin group, were isolated by laser capture microdissection and the transcriptome of the captured tissue analyzed by microarray. Gene Set Enrichment Analysis (GSEA) showed that rapamycin significantly suppressed (FDR<0.05) genes associated with oxidative phosphorylation, cell cycle progression ribosomal biogenesis and ubiquitin-mediated pro-teolysis. These results indicate that inhibition of mTOR signaling early in the process of hepatic carcinogenesis can have a persistent, anti-growth effect on gene expression. Disclosures: The following people have nothing to disclose: Adeola O. Adebayo, Heather Francois-Vaughn, Kate E. Brilliant, Philip A. Gruppuso, Jennifer A.

36 The in vivo functions

36 The in vivo functions SAR245409 datasheet of a few SNX genes have been investigated. For example, SNX1 and 2 have been knocked out in the mouse, and mice lacking either one of them are viable and fertile. However, the double-knockout mice die at midgestation, which complicates the detailed analysis of the in vivo functions of SNX1 and 2.37 SNX13 knockout mice are also embryonic lethal,38

whereas SNX27 plays essential roles during postnatal growth and survival.39 We started to investigate the in vivo functions of SNXs in the zebrafish model. We identified six SNX genes expressed in the embryonic liver and found that one of them (SNX7) was indispensable for hepatogenesis. The specification and proliferation of hepatoblasts were normal when SNX7 was blocked. However, these cells underwent extensive apoptosis during the budding stage of hepatogenesis. We concluded that an antiapoptotic activity of SNX7 was crucial for the survival

of hepatoblasts during liver budding. BMP, bone morphogenetic protein; c-FLIP, cellular FLICE-like inhibitory protein; c-FLIPL, the long form of c-FLIP; c-FLIPS, the short form of c-FLIP; CHX, cycloheximide; cp, ceruloplasmin; DAPI, 4′,6-diamidino-2-phenylindole; Wnt inhibitor dnmt, DNA methyltransferase; EGFR, epidermal growth factor receptor; FACS, fluorescence-activated cell sorting; FGFs, fibroblast growth factors; foxA3, forkhead box protein A3; gata6, GATA-binding factor 6; hdac, histone deacetylase; Hhex, hematopoietically expressed homeobox; hpf, hours postfertilization; HNF, hepatocyte nuclear factor; ifabp, intestinal fatty acid binding protein; ins, insulin; LDL, low-density lipoprotein; leg1, liver-enriched gene 1; lfabp, liver fatty acid binding protein; Mib1, mindbomb 1; MO, morpholino; mRNA, messenger RNA; mypt1, myosin phosphatase target subunit 1; PARP, poly(ADP-ribose) polymerase; P-H3, phosphorylated histone

3; Prox1, prospero homeobox protein 1; RA, selleck inhibitor retinoic acid; RT-PCR, reverse-transcription polymerase chain reaction; siRNA, short interfering RNA; SNX, sorting nexin; TGF-β, transforming growth factor beta; TNFα, tumor necrosis factor alpha; tomm22, translocase of outer mitochondrial membrane 22; try; trypsin; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling; uhrf1, ubiquitin-like protein containing PHD and Ring finger domains-1; vps18, vacuolar protein sorting protein 18; WT, wild type. Detailed protocols, including zebrafish manipulation, cell culture and short interfering RNA (siRNA) treatment, immunostaining, fluorescence-activated cell sorting (FACS) analysis, real-time reverse-transcription polymerase chain reaction (RT-PCR), and western blotting, can be found in the Supporting Materials and Methods. We performed a BLAST search against the zebrafish genome and EST databases, using human SNX sequences as references, and identified 38 zebrafish SNX family genes.

In this article, we will describe the assays used, as well as the

In this article, we will describe the assays used, as well as their

development, pitfalls in testing such as inter-laboratory variability and false negative/positive results, as well as some strategies for overcoming these pitfalls and potential alternative test approaches. The inter-laboratory coefficient of variation often approaches (and sometimes exceeds) 50%, as evidenced by various external quality assessment groups, and this variability has not improved over recent years. Additional important considerations include appropriate interpretation of test results, repeat testing for confirmation, Selleck EPZ6438 and assessment of recovery as part of the diagnostic process. The Bethesda assay (BA) for factor(F)VIII inhibitors, described in 1975 by Kasper and colleagues [1], was the first method yielding

an acceptable degree of standardization, using normal pooled plasma (NPP) as MI-503 datasheet FVIII source and imidazole buffer as reference sample. The BA replaced the Oxford assay [2], which lacked reliability as it used FVIII concentrate (cryoprecipitate) as FVIII source. Since the late 1980s, inhibitor assays were performed with increasing frequency, largely driven by multicentre studies of new purified and recombinant FVIII products. The BA soon appeared to be rather non-specific, yielding many false positive results [3]. Its sensitivity and specificity were improved

by modification in the Nijmegen assay (NA), by buffering the NPP (BNPP) and replacing the imidazole buffer with inhibitor-free selleck chemicals llc FVIII-deficient plasma as reference sample [4]. Additional recommendations, including heating of test and reference plasma to remove residual FVIII [5], the appropriate use of FVIII-deficient reference plasma [6], the use of 4M imidazole solution instead of solid imidazole for buffering the incubation mixture and the determination of residual FVIII activity with chromogenic substrates, continue to improve the sensitivity and specificity of the assay. Nevertheless, inter-laboratory coefficients of variation (CV) approaching (and sometimes exceeding) 50% are evidenced by various EQA groups, inclusive of ECAT (External quality Control of Assays and Tests) Foundation, NEQAS (UK National External Quality Assurance Scheme) and RCPA (Royal College of Pathologists of Australasia) (Fig. 1) [7-10]. This variability has not improved over the years. Reasons for such high variability largely derive from assay variability, which can occur at any stage within the assay, including choice and source of laboratory reagents, buffering (yes, no, how), NPP, etc.

In this article, we will describe the assays used, as well as the

In this article, we will describe the assays used, as well as their

development, pitfalls in testing such as inter-laboratory variability and false negative/positive results, as well as some strategies for overcoming these pitfalls and potential alternative test approaches. The inter-laboratory coefficient of variation often approaches (and sometimes exceeds) 50%, as evidenced by various external quality assessment groups, and this variability has not improved over recent years. Additional important considerations include appropriate interpretation of test results, repeat testing for confirmation, LDK378 in vivo and assessment of recovery as part of the diagnostic process. The Bethesda assay (BA) for factor(F)VIII inhibitors, described in 1975 by Kasper and colleagues [1], was the first method yielding

an acceptable degree of standardization, using normal pooled plasma (NPP) as SCH727965 FVIII source and imidazole buffer as reference sample. The BA replaced the Oxford assay [2], which lacked reliability as it used FVIII concentrate (cryoprecipitate) as FVIII source. Since the late 1980s, inhibitor assays were performed with increasing frequency, largely driven by multicentre studies of new purified and recombinant FVIII products. The BA soon appeared to be rather non-specific, yielding many false positive results [3]. Its sensitivity and specificity were improved

by modification in the Nijmegen assay (NA), by buffering the NPP (BNPP) and replacing the imidazole buffer with inhibitor-free find more FVIII-deficient plasma as reference sample [4]. Additional recommendations, including heating of test and reference plasma to remove residual FVIII [5], the appropriate use of FVIII-deficient reference plasma [6], the use of 4M imidazole solution instead of solid imidazole for buffering the incubation mixture and the determination of residual FVIII activity with chromogenic substrates, continue to improve the sensitivity and specificity of the assay. Nevertheless, inter-laboratory coefficients of variation (CV) approaching (and sometimes exceeding) 50% are evidenced by various EQA groups, inclusive of ECAT (External quality Control of Assays and Tests) Foundation, NEQAS (UK National External Quality Assurance Scheme) and RCPA (Royal College of Pathologists of Australasia) (Fig. 1) [7-10]. This variability has not improved over the years. Reasons for such high variability largely derive from assay variability, which can occur at any stage within the assay, including choice and source of laboratory reagents, buffering (yes, no, how), NPP, etc.

However, polyps located at duodenal bulb near to pylorus can not

However, polyps located at duodenal bulb near to pylorus can not be visualized completely by forward endoscopic view, unless the retroflexion view technique is performed. Endoscopic resection of this type

of polyps by using retroflexion technique in duodenum is difficult, due to the operation space is limited and narrow. We report here a case of Endoscopic resection of a polyp located at duodenal bulb near to pylorus with retroflexion technique. Methods: Using endoscopic retroflexion technique to resect a polyp located at duodenal bulb near to pylorus. Results: The lesion was completely selleck screening library resected with no complication. Conclusion: Retroflexion technique is an effective method for the resction of polyps located at duodenal bulb near to pylorus. Key Word(s): 1. Retroflexion; 2. Endoscopic resection; CAL-101 datasheet 3. duodenal polyps; Presenting Author: TAO LIU Additional Authors: HAOXUAN ZHENG, BO JIANG Corresponding Author: BO JIANG Affiliations: Department of Gastroenterology, Nanfang Hospital, Southern Medical University; Department of Gastroenterology, Nanfang Hospital, Southern Medical University Objective: Several advanced imaging techniques have been developed to improve differentiation of gastrointestinal lesions. As a precancerous lesion, atrophic gastritis should be diagnosed

and under surveillance. Confocal laser endomicroscopy (CLE) allows real-time in-vivo microscopic imaging of tissue. Narrow band imaging (NBI) and Chromoendoscopy can also diagnosis atrophic gastirtis. This study assessed the accuracy, sensitivity and specificity of those advanced techniques for diagnosis of atrophic gastritis. Methods: Consecutive patients were recruited. Each patient underwent examinations of NBI

and Chromoendoscopy, as well as CLE. Four sites of a stomach in every patient were chose to be examined by three endoscopies. Those sites were the lesser curvature of gastric antrum, the greater curvature of of gastric antrum, the middle of lesser curvature of gastric corpus and selleck compound the the middle of greater curvature of gastric corpus. During NBI and Chromoendoscopy, four sites in every patient were diagnosed for surface pit pattern. Type C, type D and type E were diagnosed of atrophic gastritis. During CLE, four sites in every patient were diagnosed for the criteria: dilated openings of gastric pits, the numbers of gastric pits reduced, exist of goblet cells or absorptive cells. Biopsies were taken from four sites after each patient examined by three endoscopies. Histopathology diagnosis served as the gold standard. Results: A total of 69 patients were in included, which contained 25 atrophic gastritis diagnosed by histopathology. The accuracy, sensitivity, and specificity of CLE were 94.2%, 92%, and 95.5%, whereas that of NBI were 75.3%, 80%, and 72.7%, and chromoendoscopy were 79.7%, 88%, and 75.

However, polyps located at duodenal bulb near to pylorus can not

However, polyps located at duodenal bulb near to pylorus can not be visualized completely by forward endoscopic view, unless the retroflexion view technique is performed. Endoscopic resection of this type

of polyps by using retroflexion technique in duodenum is difficult, due to the operation space is limited and narrow. We report here a case of Endoscopic resection of a polyp located at duodenal bulb near to pylorus with retroflexion technique. Methods: Using endoscopic retroflexion technique to resect a polyp located at duodenal bulb near to pylorus. Results: The lesion was completely buy STI571 resected with no complication. Conclusion: Retroflexion technique is an effective method for the resction of polyps located at duodenal bulb near to pylorus. Key Word(s): 1. Retroflexion; 2. Endoscopic resection; selleck 3. duodenal polyps; Presenting Author: TAO LIU Additional Authors: HAOXUAN ZHENG, BO JIANG Corresponding Author: BO JIANG Affiliations: Department of Gastroenterology, Nanfang Hospital, Southern Medical University; Department of Gastroenterology, Nanfang Hospital, Southern Medical University Objective: Several advanced imaging techniques have been developed to improve differentiation of gastrointestinal lesions. As a precancerous lesion, atrophic gastritis should be diagnosed

and under surveillance. Confocal laser endomicroscopy (CLE) allows real-time in-vivo microscopic imaging of tissue. Narrow band imaging (NBI) and Chromoendoscopy can also diagnosis atrophic gastirtis. This study assessed the accuracy, sensitivity and specificity of those advanced techniques for diagnosis of atrophic gastritis. Methods: Consecutive patients were recruited. Each patient underwent examinations of NBI

and Chromoendoscopy, as well as CLE. Four sites of a stomach in every patient were chose to be examined by three endoscopies. Those sites were the lesser curvature of gastric antrum, the greater curvature of of gastric antrum, the middle of lesser curvature of gastric corpus and selleck chemicals the the middle of greater curvature of gastric corpus. During NBI and Chromoendoscopy, four sites in every patient were diagnosed for surface pit pattern. Type C, type D and type E were diagnosed of atrophic gastritis. During CLE, four sites in every patient were diagnosed for the criteria: dilated openings of gastric pits, the numbers of gastric pits reduced, exist of goblet cells or absorptive cells. Biopsies were taken from four sites after each patient examined by three endoscopies. Histopathology diagnosis served as the gold standard. Results: A total of 69 patients were in included, which contained 25 atrophic gastritis diagnosed by histopathology. The accuracy, sensitivity, and specificity of CLE were 94.2%, 92%, and 95.5%, whereas that of NBI were 75.3%, 80%, and 72.7%, and chromoendoscopy were 79.7%, 88%, and 75.

However, polyps located at duodenal bulb near to pylorus can not

However, polyps located at duodenal bulb near to pylorus can not be visualized completely by forward endoscopic view, unless the retroflexion view technique is performed. Endoscopic resection of this type

of polyps by using retroflexion technique in duodenum is difficult, due to the operation space is limited and narrow. We report here a case of Endoscopic resection of a polyp located at duodenal bulb near to pylorus with retroflexion technique. Methods: Using endoscopic retroflexion technique to resect a polyp located at duodenal bulb near to pylorus. Results: The lesion was completely Alpelisib ic50 resected with no complication. Conclusion: Retroflexion technique is an effective method for the resction of polyps located at duodenal bulb near to pylorus. Key Word(s): 1. Retroflexion; 2. Endoscopic resection; click here 3. duodenal polyps; Presenting Author: TAO LIU Additional Authors: HAOXUAN ZHENG, BO JIANG Corresponding Author: BO JIANG Affiliations: Department of Gastroenterology, Nanfang Hospital, Southern Medical University; Department of Gastroenterology, Nanfang Hospital, Southern Medical University Objective: Several advanced imaging techniques have been developed to improve differentiation of gastrointestinal lesions. As a precancerous lesion, atrophic gastritis should be diagnosed

and under surveillance. Confocal laser endomicroscopy (CLE) allows real-time in-vivo microscopic imaging of tissue. Narrow band imaging (NBI) and Chromoendoscopy can also diagnosis atrophic gastirtis. This study assessed the accuracy, sensitivity and specificity of those advanced techniques for diagnosis of atrophic gastritis. Methods: Consecutive patients were recruited. Each patient underwent examinations of NBI

and Chromoendoscopy, as well as CLE. Four sites of a stomach in every patient were chose to be examined by three endoscopies. Those sites were the lesser curvature of gastric antrum, the greater curvature of of gastric antrum, the middle of lesser curvature of gastric corpus and learn more the the middle of greater curvature of gastric corpus. During NBI and Chromoendoscopy, four sites in every patient were diagnosed for surface pit pattern. Type C, type D and type E were diagnosed of atrophic gastritis. During CLE, four sites in every patient were diagnosed for the criteria: dilated openings of gastric pits, the numbers of gastric pits reduced, exist of goblet cells or absorptive cells. Biopsies were taken from four sites after each patient examined by three endoscopies. Histopathology diagnosis served as the gold standard. Results: A total of 69 patients were in included, which contained 25 atrophic gastritis diagnosed by histopathology. The accuracy, sensitivity, and specificity of CLE were 94.2%, 92%, and 95.5%, whereas that of NBI were 75.3%, 80%, and 72.7%, and chromoendoscopy were 79.7%, 88%, and 75.

low-risk patients: optimization using statistical models Liver T

low-risk patients: optimization using statistical models. Liver Transpl. 2006; 12(2): 231-9. 2. Kaido T, Egawa H, Tsuji H, Ashihara E, Maekawa T, Uemoto S. In-hospital mortality in adult recipients of living donor liver transplantation: experience of 576 consecutive

cases at a single center. Liver Transpl. 2009; 15(11): 1420-5. Clinical features of 450 adult LDLT recipients GRWR, graft-to-recipient weight ratio Disclosures: The following people have nothing to disclose: Murat Dayangac, Murat Akyildiz, Yalcin Erdogan, Gokhan Gungor, Yaman Tokat Introduction: Priming is essential for hepatocytes to proceed and Dorsomorphin molecular weight complete the cell cycle culminating in mitosis and replication. It remains poorly understood how hepatocytes fail to regenerate promptly in elderly animals following a two-third partial hepatectomy (PH). Since γ-aminobutyric acid (GABA) promotes hepatocytes MI-503 solubility dmso into G2 phase of cell cycle, we hypothesized that by priming old hepatocytes

to G2 phase, the liver remnants of old mice regain their regenerative capacity. Methods: We used 24-month (old) and 4-month (young) C57BL/6 mice and evaluated cell cycle distribution by immunohistochemistry for cyclin D1 (G1 phase), cyclin A (S phase), cyclin B1 (G2 phase), Ki67 and pHH3 (M phase). Results: Marked increase of Cyclin B1 positive hepatocytes was seen in aged mice following 7 days of GABA pretreatment, while control mice had mostly quiescent and Cyclin D1 positive cells (Figure 1A). The GABA treated livers regained similar mitotic activity to young controls as seen by pHH3 and Ki67 staining at 36 h after PH. The results were confirmed with western and further explored through gene expression analyses (data not shown). In a separate experiment, mice with and without GABA pre-treatment were followed daily through day 7 post PH to establish cell cycle profile by IHC. We found that Old-GABA mice had proliferative Ki-67 and see more mitotic pHH3 profiles that were similar to those of young

mice (Figure 1B). Conclusion: Our results indicate that hepatic regenerative capacity after PH in elderly mice can be restored by priming hepatocytes to G2 state prior to PH. Improvement in liver regeneration in elderly will impact quality of liver grafts from elderly donors. Disclosures: The following people have nothing to disclose: Fatima K. Rehman, Toshiyuki Hata, Zhaoyu Li, Guojun Bu, Justin H. Nguyen Introduction: MELD score (Model for End Stage Liver Disease) is universally used to priorities patients on the liver transplant waiting list. It is potentially used to predict survival as well. There has been conflicting evidence on using living donor liver transplantation (LDLT) in patients with high MELD score. We herein showing a retrospective analysis of survival data in these two categories of patients and comparing survival between LDLT and Deceased Donor liver Transplantation (DDLT) in a single center experience.

low-risk patients: optimization using statistical models Liver T

low-risk patients: optimization using statistical models. Liver Transpl. 2006; 12(2): 231-9. 2. Kaido T, Egawa H, Tsuji H, Ashihara E, Maekawa T, Uemoto S. In-hospital mortality in adult recipients of living donor liver transplantation: experience of 576 consecutive

cases at a single center. Liver Transpl. 2009; 15(11): 1420-5. Clinical features of 450 adult LDLT recipients GRWR, graft-to-recipient weight ratio Disclosures: The following people have nothing to disclose: Murat Dayangac, Murat Akyildiz, Yalcin Erdogan, Gokhan Gungor, Yaman Tokat Introduction: Priming is essential for hepatocytes to proceed and Bortezomib concentration complete the cell cycle culminating in mitosis and replication. It remains poorly understood how hepatocytes fail to regenerate promptly in elderly animals following a two-third partial hepatectomy (PH). Since γ-aminobutyric acid (GABA) promotes hepatocytes Selleckchem Palbociclib into G2 phase of cell cycle, we hypothesized that by priming old hepatocytes

to G2 phase, the liver remnants of old mice regain their regenerative capacity. Methods: We used 24-month (old) and 4-month (young) C57BL/6 mice and evaluated cell cycle distribution by immunohistochemistry for cyclin D1 (G1 phase), cyclin A (S phase), cyclin B1 (G2 phase), Ki67 and pHH3 (M phase). Results: Marked increase of Cyclin B1 positive hepatocytes was seen in aged mice following 7 days of GABA pretreatment, while control mice had mostly quiescent and Cyclin D1 positive cells (Figure 1A). The GABA treated livers regained similar mitotic activity to young controls as seen by pHH3 and Ki67 staining at 36 h after PH. The results were confirmed with western and further explored through gene expression analyses (data not shown). In a separate experiment, mice with and without GABA pre-treatment were followed daily through day 7 post PH to establish cell cycle profile by IHC. We found that Old-GABA mice had proliferative Ki-67 and selleck kinase inhibitor mitotic pHH3 profiles that were similar to those of young

mice (Figure 1B). Conclusion: Our results indicate that hepatic regenerative capacity after PH in elderly mice can be restored by priming hepatocytes to G2 state prior to PH. Improvement in liver regeneration in elderly will impact quality of liver grafts from elderly donors. Disclosures: The following people have nothing to disclose: Fatima K. Rehman, Toshiyuki Hata, Zhaoyu Li, Guojun Bu, Justin H. Nguyen Introduction: MELD score (Model for End Stage Liver Disease) is universally used to priorities patients on the liver transplant waiting list. It is potentially used to predict survival as well. There has been conflicting evidence on using living donor liver transplantation (LDLT) in patients with high MELD score. We herein showing a retrospective analysis of survival data in these two categories of patients and comparing survival between LDLT and Deceased Donor liver Transplantation (DDLT) in a single center experience.

2 chain, hypersecrete T helper 1 (Th1) and Th2 cytokines and chem

2 chain, hypersecrete T helper 1 (Th1) and Th2 cytokines and chemokines upon stimulation with an appropriate ligand, such as alpha-galactosylceramide (α-GalCer). In doing so, these iNKT cells exert considerable and promiscuous immune function, including altering immune regulation by activating dendritic cells (DCs), macrophages, NK cells, T cells, B cells, and driving the development of adaptive immunity.12, 13 iNKT cells appear to play a critical role in the regulation of several other autoimmune diseases, including type 1 diabetes, multiple sclerosis, rheumatoid arthritis, and systemic lupus erythematosus.13-17 Previously, our laboratory has proposed

that activation of iNKT cells is a critical factor in accelerating disease.18-20 To explore this issue Acalabrutinib mw in depth, we immunized C57BL/6 mice with 2-OA-BSA (bovine serum albumin) and activated their iNKT cells with

α-GalCer. We report herein that iNKT cell activation by α-GalCer leads to a profound exacerbation of portal disease in 2-OA-BSA-immunized mice, including increased AMA production, increased CD8+ T cell biliary infiltration, portal inflammation, granuloma formation, bile duct damage, and fibrosis. These results are critical and emphasize the role of innate immunity in the natural history of PBC and they further suggest mechanisms by which biliary disease becomes perpetuated in humans as well as explaining the recurrence of selleck kinase inhibitor PBC following liver transplantation in the absence of major histocompatability complex (MHC) compatibility. These data also emphasize the appearance of fibrosis, a feature thus far lacking in the other murine models of autoimmune cholangitis. 2-OA, 2-octynoic acid; α-GalCer, alpha-galactosylceramide; α-SMA, alpha-smooth muscle actin; AMAs, antimitochondrial antibodies; BSA, bovine serum albumin; CFA, complete Freund’s adjuvant; DC, dendritic cells; dnTGF-βRII, TGF-β receptor II dominant-negative; HSCs, hepatic stellate cells; iNKT, invariant natural killer T; PBC, primary biliary cirrhosis; selleck inhibitor PDC-E2, E2 subunits of the pyruvate dehydrogenase complex; TGF-β,

transforming growth factor beta. The protocol for induction of autoimmune cholangitis is similar to our previous reports.7 Briefly, female C57BL/6 mice aged 8-10 weeks were obtained from the National Laboratory Animal Center and maintained in the Animal Center of the College of Medicine, National Taiwan University. Mice were intraperitoneally immunized with 2-OA-BSA (100 μg) in the presence of complete Freund’s adjuvant (CFA, Sigma-Aldrich, St. Louis, MO) and subsequently boosted at weeks 2, 4, 6, 8, and 10 with 2-OA-BSA and incomplete Freund’s adjuvant (IFA, Sigma-Aldrich). Two μg of α-GalCer (Alexis, San Diego, CA) was diluted in phosphate-buffered saline (PBS) and injected intravenously 1 day before first, second, and third 2-OA-BSA immunizations (group name: α-GC/CFA/2-OA).