Reverse transfections of RNA oligoribonucleotides were performed with Lipofectamine-RNAiMAX (Invitrogen). A total of 50 nM of RNA duplex or 200 check details nM of miRNA inhibitor

were used for each transfection. Cotransfections of RNA duplex with plasmid DNA were performed with Lipofectamine 2000 (Invitrogen). HEK293T cells were transfected with plasmids by calcium phosphate precipitation. Packaging of the retroviral expression vectors and infections of the target cells were performed as described in the Supporting Materials and Methods. Thirty-six hours after the reverse transfections with RNA oligonucleotides, the tumor cells were washed with 1× phosphate-buffered saline (PBS) and were cultured in SFM for 12 hours; tumor cell–conditioned medium (TCM) was collected subsequently as described in Supporting Materials and Methods.

For all experiments, TCM loading was adjusted according to the number of live cells in each sample. Assays to determine the effects of tumor cells or TCM on the migration or capillary tube formation of HUVECs were performed as described in Supporting Materials and Methods. The migration and invasion of tumor cells were analyzed in 24-well Boyden chambers with 8-μm pore size polycarbonate membranes (Corning, NY). For invasion assays, the membranes were coated with Matrigel (3432-005-01, R&D Systems, MN) to form matrix barriers. All experimental procedures involving animals were performed in accordance with the Guide for the Care and Use of Laboratory Animals (National Institutes Venetoclax order of Health publication nos. 80-23, revised 1996) and according to the institutional ethical guidelines for animal experiments. For xenograft implantation experiments, 2 × 106 QGY-miR-195-LUC cells were resuspended in 25 μL of PBS/Matrigel (1:1) and were inoculated under the capsule of the left hepatic lobe of male BALB/c nude mice. miR-195 expression was silenced by administering drinking water

that was supplemented with 10% sucrose and 2 mg/mL of doxycycline MycoClean Mycoplasma Removal Kit (Clontech). Bioluminescence imaging and tumor dissection were performed as described in Supporting Materials and Methods. QGY-7703 cells in a 48-well plate were cotransfected with 50 nM of miR-195 or NC duplex, 10 ng of pRL-TK (Promega, Madison, WI), and 50 ng of firefly luciferase reporter plasmid that contained either the wild-type or mutant 3′UTR of the target gene. Forty-eight hours after the transfections, the cell lysates were applied to luciferase assay as described.[3] The levels of VEGF in the TCM were detected using enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems) as instructed by the manufacturer. Lysates from QGY-7703 cells were incubated with glutathione-Sepharose bead–immobilized glutathione S-transferase (GST) or GST-PAK. Next, the bead-bound proteins were solubilized in sodium dodecyl sulfate (SDS) buffer and analyzed by immunoblotting.

Distribution of HLA ligands C1 (HLA-C, 80N), C2 (80K), Bw4 (HLA-B, 80I or T and HLA-A*2301, 2402 and 3201) and HLA-A3/A11 did not differ significantly between DILI patients and controls. The most frequent receptor-ligand combinations in the DILI patients were 2DL3 + C1 (67%) and 3DL1 + Bw4 (67%), while 2DL1 + C2 (69%) and 3DL1 + Bw4 (69%) predominated in the controls. In contrast, 3DS1+Bw4 was the least frequent receptor-ligand combination in DILI (9%) and controls (11%). Conclusion: This is to our knowledge the first KIR association analysis in DILI patients. However, our AC DILI cohort presented KIR gene distributions similar to the controls, which were comparable to previously

reported KIR data from RGFP966 CDK inhibitor drugs ethnically similar cohorts. Furthermore, the analysed

KIR receptor–HLA ligand combinations do not appear to play a major role in AC DILI development. The complete ligand repertoire is however not elucidated and a potential role for KIR in AC DILI should not yet be dismissed. Funding: PI-0239-2012 SAS, FIS PI12-00378, AC-0073-2013 SAS, CIBERehd by ISCIII Disclosures: Manuel Romero-Gomez – Advisory Committees or Review Panels: Roche Farma,SA., MSD, S.A., Janssen, S.A., Abbott, S.A.; Grant/Research Support: Ferrer, S.A. The following people have nothing to disclose: C. Stephens, Antonia More-no-Casares, MAngel López-Nevot, Miren García-Cortés, I. Medina-Cáliz, Hacibe Hallal, German Soriano, Francisco. Ruiz-Cabello, M. I. Lucena, Raul J. Andrade [Background and Aim] The recent global increase in the incidence of metabolic syndrome has also impacted its hepatic manifestation

in the form of an increased prevalence of non-alcoholic fatty liver disease (NAFLD). It is known that liver metabolism is regulated by a ‘metabolic highway’ mediated by the autonomic nervous system. The component neurons are distributed within the liver, extending from the portal area, and then gradually branching to form a fine network around the portal area. However, the precise mechanism by which this regulatory system operates is still poorly understood. Therefore, the aim of the present study was to examine the role of the autonomic nervous system in the liver using immunohistochemistry, and to clarify the association between these nerves and a variety of liver diseases. [Methods] First, we evaluated Adenylyl cyclase the autonomic nervous system in the liver after transplantation, which in theory should lead to intrahepatic neuron atrophy. As neuron markers, we evaluated changes in S-100 or N-CAM immunostaining over time (n=90). Specimens of normal liver obtained at surgery for metastatic liver cancer were used as immunostaining controls (n=5). Also, we evaluated a diverse group of liver diseases (NASH n=18, chronic hepatitis B n=10, chronic hepatitis C n=10) to evaluate whether these diseases show differences in the ratio of positivity for neuron markers.

devised mechanistic experiments in the methionine-choline deficient Selleckchem Cabozantinib (MCD) model of NASH. Immunization with malonyldialdehyde-adducted bovine serum albumin before the deficient diet worsened all severity parameters examined in animals subjected to this model, except for hepatic triglyceride content. In immunized MCD animals, there was a recruitment of T lymphocytes and natural killer T cells in the liver with an increased hepatic content of IL-15 and osteopontin. Furthermore, the researchers showed that depletion of CD4+ T lymphocytes improved the lesions in immunized animals.

This work complements remarkably the clinical observations and suggests that NASH might be an immunologic disease. (Hepatology

2014;59:886–897.) Liver metastases of nonliver tumors often do not catch the attention of hepatologists. However, they are much more frequent than primary liver cancer and are an underexplored field of research. In an astonishing article, Turtoi et al. performed matrix-assisted laser desorption/ionization image-guided proteomic analysis of colorectal cancer liver metastases. The reaserchers consistently observed a different distribution of proteins in three spatially distinct zones: the center of the metastasis; the rim of the metastasis; and the peritumoral region. They identified several extracellular proteins selleck compound expressed only in the tumoral stroma, which have potential as therapeutic targets. These results are based on a small number of lesions, but they illustrate the potential

of this approach; still currently very sophisticated, this technology will become more accessible and widely used. (Hepatology 2014;59:924–934.) “
“Acute pancreatitis is an inflammatory response to pancreatic injury., The response can be mild and local, or extend to peripancreatic and systemic inflammation. The systemic inflammatory response can also lead to vascular leak, pulmonary edema, hypovolemia hypotension and shock, and ischemic injury to the pancreas, kidney and intestines. The most common etiologies of acute pancreatitis are gallstones, alcoholism, and idiopathic. Treatment is initially aimed at fluid resuscitation, Cytidine deaminase support of organ dysfunction, and in severe cases enteral feeding. Therapeutic endoscopy may be needed to dislodge an impacted gallstone to treat bacterial cholangitis. Treatment is supportive, and a plan to prevent recurrence should be implemented before discharge. “
“A 27-year-old man with poorly controlled type 1 diabetes mellitus (average hemoglobin A1c of 15%) presented with a 1-week history of progressive pressure-like right upper abdominal discomfort associated with early satiety and nausea. On physical exam, he had firm hepatomegaly extending into the right pelvis.

Also, early recognition is key to preserving unnecessary operation or other intervention. Key Word(s): 1. liver cirrhosis; 2. intramuscular hematoma; 3. outcome; 4. treatment Table 1 Intramuscular hematoma in patients with liver cirrhosis Case Age Gender Etiology Location Treatment Outcom Child-Pugh score Case A–C: Our Patients; Case 1–8 Previous repoted patients; HCV: Hepatitis C virus;TAE: Transcatheter arterial embolism Presenting Author: SULAIMAN BUDIMAN SUJATMIKA Additional Authors: RINO A GANI, C RINALDI A LESMANA, MARCELLUS SIMADIBRATA, Atezolizumab clinical trial ENING KRISNUHONI, ALI SULAIMAN Corresponding Author: BUDIMAN SUDJATMIKA Affiliations:

Cipto Mangunkusumo Hospital, Cipto Mangunkusumo Hospital, Cipto Mangunkusumo Hospital, Cipto Mangunkusumo Hospital, Cipto LGK-974 solubility dmso Mangunkusumo Hospital Objective: Chronic hepatitis B (CHB) infection is still a major problem in most Asian countries. The role of seromarkers such as quantitative HBV DNA and quantitative HBsAg might not predict accurately the intrahepatic viral activity. Intrahepatic cccDNA and Pregenomic RNA measurement is not practical in clinical practice. Quantification of intrahepatic HBsAg might give a better prediction in clinical practice since the liver biopsy is also a common procedure for the liver

condition assessment. To know the clinical significance of intrahepatic HBsAg measurement in correlation with quantitative HBsAg serum in CHB patients who received oral antiviral therapy. Methods: This was a retrospective cohort study using liver specimens and blood samples from 26 CHB patients who underwent oral antiviral therapy for one year (2010–2011) from two big referral hospitals (Cipto Mangunkusumo Hospital

and Medistra Hospital). Quantification of HBsAg particles in the liver specimens was done by immunohistochemistry staining. Other data was obtained from the patient’s database. Statistical analysis was done using SPSS ver. 17. Results: There was a weak correlation between quantitative HBsAg serum with intrahepatic HBsAg particles before and after one year ADP ribosylation factor oral antiviral therapy (r = −0.20, p .249; r = 0.15, p .433). Quantitative HBsAg serum was decreased after one year oral antiviral therapy, however there was no mean difference in intrahepatic HBsAg particles after one year oral antiviral therapy (p .468). Conclusion: Intrahepatic viral activity might not be accurately predicted by seromarkers as quantitative HBsAg serum is more related to viral load but intrahepatic HBsAg particles reflects more intrahepatic viral activity that many factors could influence this condition. Further study is needed with larger samples to confirm these findings.

“
“Hepatoblastoma (HB) is the most common primary liver tumor in children. Mutations in the β-catenin

gene that lead to constitutive activation of the Wnt pathway have been detected in a large proportion of HB tumors. To identify novel mutations in HB, we performed whole-exome sequencing of six paired HB tumors and their corresponding lymphocytes. This identified 24 somatic nonsynonymous mutations in 21 genes, many of which were novel, including three novel mutations targeting the CTNNB1 (G512V) and CAPRIN2 (R968H/S969C) genes in the Wnt pathway, selleck compound and genes previously shown to be involved in the ubiquitin ligase complex (SPOP, KLHL22, TRPC4AP, and RNF169). Functionally, both the CTNNB1 (G512V) and CAPRIN2 (R968H/S969C) were

observed to be gain-of-functional mutations, and the CAPRIN2 (R968H/S969C) was also shown to activate the Wnt pathway in HB cells. These findings suggested the activation of the Wnt pathway in HB, which was confirmed by immunohistochemical staining of the β-catenin in 42 HB tumors. We further used short hairpin RNA (shRNA)-mediated interference to assess the effect see more of 21 mutated genes on HB cell survival. The results suggested that one novel oncogene (CAPRIN2) and three tumor suppressors (SPOP, OR5I1, and CDC20B) influence HB cell growth. Moreover, we found that SPOP S119N is a loss-of-function mutation in HB cells. We finally demonstrated that one of the mechanisms by which SPOP inhibits HB cell proliferation is through regulating CDKN2B expression. Conclusion: These results extend the landscape of genetic alterations in HB and highlight the dysregulation of Wnt and ubiquitin pathways in HB tumorigenesis. (Hepatology 2014;;60:1686–1696) “
“Drug-induced liver injury (DILI) is a leading cause of drug failure in clinical trials and a major reason for drug withdrawals from the market. Although there is evidence that dosages of ≥100 mg/day

are associated with increased risk for hepatotoxicity, many drugs are safe at such dosages. There is an unmet need to Niclosamide predict risk for DILI more reliably, and lipophilicity might be a contributing factor. We analyzed the combined factors of daily dose and lipophilicity for 164 US Food and Drug Administration–approved oral medications and observed high risk for hepatotoxicity (odds ratio [OR], 14.05; P < 0.001) for drugs given at dosages ≥100 mg/day and octanol-water partition coefficient (logP) ≥3. This defined the “rule-of-two.” Similar results were obtained for an independent set of 179 oral medications with 85% of the rule-of-two positives being associated with hepatotoxicity (OR, 3.89; P < 0.01). Using the World Health Organization’s Anatomical Therapeutic Chemical classification system, the rule-of-two performed best in predicting DILI in seven therapeutic categories.

In contrast, metformin was associated with a decreased risk for liver cancer.

Consistent with previous in vitro studies on TZDs which showed antiproliferation and prodifferentiation effects, our data have provided an association between the clinical use of TZDs and a reduced risk for several cancer incidences, in particular liver cancer. The association became stronger when the duration of TZD use was longer and the dosage was higher. Rosiglitazone, but not pioglitazone, was associated with a significantly reduced risk for colorectal cancer. STA-9090 No association between both TZDs and lung and bladder cancer was observed. Previous reports on the association between TZD use and cancer incidence have been inconsistent. The report from the data obtained from the Veterans Integrated Services Network 16 (VISN 16) cohort of 87,678 individuals showed a 33% reduction in lung cancer risk among TZD users compared with nonusers (relative risk: 0.65, 95% CI: 0.51-0.87). However, as rosiglitazone and pioglitazone were combined, the risk reduction

for colorectal PF-562271 in vitro cancer did not reach statistical significance. 18 In contrast, the present study results did not show a decreased risk for lung cancer. Although numerous in vitro studies support the protective effect of TZDs in lung cancer, the specific tissue or type of cancer and its stage might contribute to the efficacy or failure of TZDs as antineoplastic agents. 19, 20, 24-29 Because the risk factors, genetic expressions, and pharmaceutical responses of lung cancer of the Taiwanese differ significantly from those in the Western countries, there might also be a differential response to TZDs. 30 On the contrary, our analysis showed a protective effect of rosiglitazone on colorectal cancers, which was not evident in the VISN 16 cohort. In animal studies, PPAR-γ agonists inhibited tumor growth

and colon carcinogenesis through induction of apoptosis and suppression of the cell cycle. 31-34 The current study, to the best of our knowledge, provides the first evidence that rosiglitazone but not pioglitazone might reduce Masitinib (AB1010) the risk of colorectal cancer. It is initially surprising that both pioglitazone and rosiglitazone are associated with a reduced risk for liver cancer. Hepatocellular carcinoma, one of the most incident, prevalent, and lethal malignancies in Taiwan, is regarded as a late-stage sequel of chronic infection of hepatitis B and C. 35, 36 With only a few exceptions, the development of hepatocellular carcinoma almost exclusively follows the sequence of chronic hepatic inflammation, cirrhosis of the liver, repair and regeneration of hepatic cells, and then carcinogenesis. 37 This might explain the finding that risk reduction was more evident in the patients with chronic liver disease. Despite the concern that physicians may preferentially prescribe TZDs to patients with better liver function (i.e.

Endoscopic drainage was done in 16 patients; 2 resolved, 11 had resolving WON and 2 had persistent WON. 24 patients required

surgery in total and 12 patients expired. Conclusion: Majority of fluid collections are acute necrotic Peptide 17 mouse collections (ANC) and majority of them developed WON. Pseudocyst occurs extremely rare in the natural history of acute pancreatitis. Infections and need for intervention were seen predominantly in patients with ANC and half of them can be managed conservatively. Key Word(s): 1. Acute pancreatitis; 2. Fluid collections; 3. Pseudocyst; 4. WON; Presenting Author: MALAY SHARMA Additional Authors: CHITRANSHU VASHISHTHA Corresponding Author: MALAY SHARMA Affiliations: Jaswant Rai Speciality Hospital; Institute of Liver & Biliary Sciences Objective: Acute Pancreatitis (AP) can occur due to presence of impacted small stones in prepapillary area. These stones can also migrate into pancreatic duct (PD). The aim of the study was to determine the role of Endoscopic ultrasound (EUS) in finding prepapillary and migrated intrapancreatic stone by EUS in AP (within first 48 hrs) where history and investigations failed to reveal a cause. Methods: 280 patients

of AP admitted from September 2005 to March 2013 underwent clinical evaluation and baseline biochemistry was done. Transabdominal ultrasonogram &/or CECT of abdomen was done. Patients with first attack of pancreatitis, where aetiology was not known and in whom EUS was done during the acute episode were included for analysis. Those with previous attacks of pancreatitis or with an established aetiology after these investigations were excluded. Obeticholic Acid Results: Out of 280 patients admitted with AP, 85 fulfilled the inclusion criteria. Endoscopic ultrasound was able to suggest the etiology in 46 patients. Gallbladder

stone related disease was found in 38 cases. 9 cases had prepapillary stone of CBD origin and 5 had PD stones which had migrated from CBD. Other causes included suprapapillary CBD stone (9), microlithiasis Ponatinib solubility dmso of gall bladder (1), sludge in gall bladder (13) and microlithiasis of common bile duct (1). Conclusion: Early EUS has different therapeutic impact as compared to EUS after 4 weeks in AP. Dilated PD in AP may be due to impacted or migrated CBD stones which can be easily identified by EUS. When a stone migrates into PD it can dilate for a while but the duct becomes normal subsequently in most of the cases. EUS should be done early to manage a subgroup of cases of AP. Key Word(s): 1. Acute Pancreatittis; Presenting Author: RAJESH GUPTA Additional Authors: SUNIL SHENVI, SHRUTI HS, DEEPAK BHASIN, SURINDER RANA Corresponding Author: RAJESH GUPTA Affiliations: PGIMER Objective: Debilitating abdominal pain remains the most common presentation of chronic pancreatitis and the treatment remains challenging. This study analyzed the outcome of surgery in patients with chronic pancreatitis.

It is important to identify the major (and treatable) risks for this purpose. The duration of pretransplant abstinence, HRAR score, non-compliance, personality disorder, mental illness, lack of a stable partner,

amount of consumption of alcohol at the time of assessment, reliance on family or friends learn more for post-transplant support, tobacco consumption at the time of assessment and lack of insight into alcohol were presented in various reports.[3, 9-11] Our recent report revealed that the postoperative circumstances correlated more strongly with the incidence of postoperative alcohol relapse than the pattern of alcohol consumption before transplantation.[2] Our current study showed that pretransplant abstinence shorter than 18 months was a significant indicator for harmful relapse. We did not find that HRAR score predicted recidivism Hormones antagonist or harmful relapse. Although 6 months of abstinence did not predict recidivism or harmful relapse in this study, it successfully selected patients who

recovered from liver failure without LDLT in a single-center report in Japan.[12] Although the use of 18 months of abstinence as a transplant criterion would eliminate the majority of harmful relapse patients, it would also eliminate many patients not drinking harmfully or remaining abstinent during our follow-up period. Nevertheless, patients with pretransplant sobriety shorter than 18 months may be an appropriate target group for a more intensive alcohol rehabilitation pre- and post-transplant. Another discussion is whether non-harmful relapse is acceptable or not. From a clinical point of view, it may be acceptable because the survival rate is similar to that of abstinent patients. However, it may not be acceptable from a public point of view because of the issue of organ

donation and the shortage of organs. In other words, it may be acceptable in LDLT but not in DDLT. However, because 21 of 32 patients with recidivism fell into the category of harmful relapse, any alcohol intake should be avoided after transplantation, both in LDLT and DDLT. Because there were no significant Y-27632 2HCl pretransplant data to predict recidivism in our recent report, establishment of a good alcohol rehabilitation program is the only option. The findings of this retrospective, multicenter study are limited by several factors inherent in this type of study, including variability in documentation, differences in selection criteria and data collection, and missing data. To minimize variability, we sent a standardized collection form containing 150 questions to the transplant centers. The element of time should be taken into account in the statistical analyses because subjects have differing lengths of follow up for entire post-transplant periods and period with alcohol consumption after transplantation.

Liver biopsies were read and scored according to NASH CRN scoring system. The associations of premature menopause and the time from menopause with fibrosis severity (stage 0-4) were assessed using ordinal logistic regression models with and without adjusting for age at enrollment, race/ethnicity, waist circumference, current smoking/alcohol use, hypertension, diabetes, Rapamycin mw HOMA-IR, and hormone replacement therapy (HRT). [Results]

Mean age ± SD at menopause was 44 ± 9 yrs. 29.5% women had premature menopause. This group was younger at enrollment (55 ± 9 vs. 59 ± 7 yrs, p<0.001, t-test) and used HRT more often (32% vs. 19%, p<0.003, Chi-square test). Premature menopause was associated with a higher stage of fibrosis (p<0.05, Wil-coxon rank-sum test).

No significant differences were noted in other histologic features or above-listed clinical variables. After adjusting for age at enrollment, premature menopause was associated with an increased likelihood of having more severe fibrosis; cumulative odds ratio and 95% confidence interval (COR[95%CI]) was 1.7[1.2, 2.4] (p< 0.004), which remained similar after adjusting for the other variables. Time from menopause was also directly associated with an increased likelihood of having more severe fibrosis (COR[95%CI] for 5-year unit=1.2[1.1, 1.3], p<0.0001), which also remained the same after adjusting for the other variables. [Conclusion] Age at menopause was significantly associated with a risk of fibro-sis among post-menopausal BGJ398 women with NAFLD. This finding, together with our previous reports, underscores the significance of reproductive ADAM7 information for risk stratification among female patients with NAFLD. Disclosures: Manal F. Abdelmalek – Consulting: Islet Sciences; Grant/Research Support: Mochida Pharmaceuticals,

Gilead Sciences, NIH/NIDDK, Synageva, Genfit Pharmaceuticals Joel E. Lavine – Consulting: Merck, Crosscare, Gilead, Takeda Millenium; Grant/ Research Support: Janssen Anna Mae Diehl – Consulting: Roche; Grant/Research Support: Gilead, Genfit The following people have nothing to disclose: Jagpal S. Klair, Ju Dong Yang, Cynthia D. Guy, Ryan M. Gill, Katherine P. Yates, Aynur Unalp-Arida, Jeanne M. Clark, Ayako Suzuki Background: Experimental evidence suggests a cardiopro-tective role of heat shock proteins (HSP) in several models of acute myocardial stress. Patients with both non-alcoholic fatty liver disease (NAFLD) and coronary artery disease (CAD) have lower levels of serum HSP. HSP may stimulate autoimmune responses resulting in the production of antibodies. Aim The aim of this study is to investigate the possible role of anti-HSP auto-antibodies in reduction of HSP in patients with NAFLD, leading to increased prevalence of coronary artery disease (CAD). Methods: We prospectively enrolled 119 patients undergoing elective coronary angiography. Each patient had fasting serum, clinical data and abdominal ultrasound (US). All US were read centrally by a standard protocol.

In our population, it also seems to have a greater influence on treatment response. IL28B’s mechanism of action is likely effected through ISG expression. These results indicate that patient’s native IL28B genotype has a greater influence than donor genotype on relevant ISG expression, and consequently on treatment response. Disclosures: Richard Gilroy – Advisory Committees

or Review Panels: FDA GIDAC; Speaking and Teaching: Salix, Vertex, Gilead The following people have nothing to disclose: Zoe Raglow, Chuanghong Wu, Winston PKC inhibitor Dunn, Yu-Jui Y. Wan BACKGROUND & AIMS MicroRNAs (miRNAs) are an important class of small non-coding RNA molecules that bind to their complementary sequence on their target mRNAs, resulting in translational repression. MiRNAs play major

roles in development, metabolism, infection, and cancer. The diagnosis of cholangiocarcinoma is sometimes difficult due to the lack of a specific tumor marker and limited sampling for histological evaluation. In this study, we analyzed miRNA expression in bile to identify predictive miRNAs for hepatobiliary malignancy. METHODS MiRNAs were obtained from bile samples taken from 25 patients with benign hepatobiliary disease and 34 patients with malignant hepatobiliary disease, including cholangiocarcinoma and gallbladder cancer. Expression of 328 miRNAs was determined with the TaqMan real-time PCR detection system using a MicroRNA Assays Human Panel. MiRNAs were functionally ZD1839 in vitro evaluated in cholangiocarcinoma cell lines (Huh28, Hucct1, and KMBC) following the overexpression or knocking down of specific miRNAs using mimic-miRNA or anti-miRNA. RESULTS Among the 328 miRNAs tested, 16 were differentially expressed between the bile of patients with malignant hepatobiliary disease and those

with benign hepatobiliary to disease (p<0. 05). From these 16 miRNAs, we focused on the two—miR-451 and miR-486—that were most significantly increased in malignant hepatobiliary disease. The area under the curve of the receiver operating characteristic curve for the prediction of malignant hepatobiliary disease using these miRNAs was 0. 85, which was equivalent to that of the tumor marker CA19-9. Univariate analysis using multiple clinical parameters, including tumor markers (CEA and CA19-9) and a liver function test, revealed serum ALT and T-Bil, miR-451, and miR-486 as significant variables. Multivariate analysis identified CEA, CA19-9, and miR-486 as significant variables. Functional analysis of these miRNAs showed that miR-451 had tumor suppressive and anti-inflammatory properties by decreasing the expression of PDGFRα, KRT7, IL1, and IL6, while miR486 was tumorigenic by increasing the expression of PDGFRα, N-cadherin, and p38MAPK. CONCLUSIONS We identified two predictive miRNAs, miR-451 and −486, in bile for the diagnosis of malignant hepatobiliary disease.