16, 17 With this limited genome, HCV replicates in hepatocytes by relying on cellular systems, thereby co-opting cellular proteins in its life cycle. To date, HCV particles have been found to contain more than 10

host lipoproteins.11, 12 Incorporation of these host proteins into HCV virions may not be R788 ic50 random, as other enveloped viruses selectively obtain host proteins. For example, HIV-1 selectively acquires more than 40 host proteins, but excludes some proteins such as CD4, CD45, CXCR4, CCR3, and CCR5, which are all highly expressed on HIV-1-infected cells.20 It is believed that HIV-1 acquires biologically functional RCA proteins during viral budding at the plasma membrane. HCV, however, may acquire CD59 while budding through the membranes of intracellular organelles rather than at the plasma membrane because HCV may exit the cells by way of a secretory pathway.21 FACS and western blot data in this study showed that human hepatocytes expressed high levels of intracellular and surface CD59 without a difference in their molecular weights (Fig. 1), thereby providing a possible

source for HCV to encounter and obtain CD59 in intracellular organelles such as the ER. Identifying these interactions is critical for understanding the life cycle of HCV, and may yield novel targets for development of therapeutic strategies. To date, abrogation of RCA function to regain antivirus Ab activity against enveloped viruses has only been check details tested in vitro in artificial environments such as GVB systems.2, 5, 6, 8 These systems provide optimized conditions for complement activation in vitro, but have no clinical relevance because they do not adequately replicate in vivo conditions. In this study, CD59 blockers were directly added into patient plasma to abrogate the function of CD59 on the patient’s own viral particles,

resulting in destruction of primary virions. The enhanced virolysis was likely triggered by ADCML, as all six individuals chronically infected with HCV showed high titers of anti-E2 Abs and potent complement activity. ADCML efficacy, however, significantly varied among these samples, which may be affected by many factors including the nature of the host immune response, HCV virological features, and patient profiles, medchemexpress because they all affect the outcome of HCV infection. For example, HCV from patient Pt42 was one of the most resistant viruses to the ADCML. Accordingly, this patient had the lowest anti-HCV E2 Ab titer among all six patients examined. However, our sample size is too small to analyze the correlation between the Ab titer and virolysis. In addition, anti-HCV E1 Abs in plasma samples from these patients were not titrated. Thus, further investigations with large clinical samples are required to analyze the correlation of anti-HCV E1/E2 Ab titers and virolysis efficacy.

16, 17 With this limited genome, HCV replicates in hepatocytes by relying on cellular systems, thereby co-opting cellular proteins in its life cycle. To date, HCV particles have been found to contain more than 10

host lipoproteins.11, 12 Incorporation of these host proteins into HCV virions may not be this website random, as other enveloped viruses selectively obtain host proteins. For example, HIV-1 selectively acquires more than 40 host proteins, but excludes some proteins such as CD4, CD45, CXCR4, CCR3, and CCR5, which are all highly expressed on HIV-1-infected cells.20 It is believed that HIV-1 acquires biologically functional RCA proteins during viral budding at the plasma membrane. HCV, however, may acquire CD59 while budding through the membranes of intracellular organelles rather than at the plasma membrane because HCV may exit the cells by way of a secretory pathway.21 FACS and western blot data in this study showed that human hepatocytes expressed high levels of intracellular and surface CD59 without a difference in their molecular weights (Fig. 1), thereby providing a possible

source for HCV to encounter and obtain CD59 in intracellular organelles such as the ER. Identifying these interactions is critical for understanding the life cycle of HCV, and may yield novel targets for development of therapeutic strategies. To date, abrogation of RCA function to regain antivirus Ab activity against enveloped viruses has only been selleck compound tested in vitro in artificial environments such as GVB systems.2, 5, 6, 8 These systems provide optimized conditions for complement activation in vitro, but have no clinical relevance because they do not adequately replicate in vivo conditions. In this study, CD59 blockers were directly added into patient plasma to abrogate the function of CD59 on the patient’s own viral particles,

resulting in destruction of primary virions. The enhanced virolysis was likely triggered by ADCML, as all six individuals chronically infected with HCV showed high titers of anti-E2 Abs and potent complement activity. ADCML efficacy, however, significantly varied among these samples, which may be affected by many factors including the nature of the host immune response, HCV virological features, and patient profiles, MCE公司 because they all affect the outcome of HCV infection. For example, HCV from patient Pt42 was one of the most resistant viruses to the ADCML. Accordingly, this patient had the lowest anti-HCV E2 Ab titer among all six patients examined. However, our sample size is too small to analyze the correlation between the Ab titer and virolysis. In addition, anti-HCV E1 Abs in plasma samples from these patients were not titrated. Thus, further investigations with large clinical samples are required to analyze the correlation of anti-HCV E1/E2 Ab titers and virolysis efficacy.

For approved compounds, combination of AL-335 with the NS3/4A protease inhibitor, Copanlisib in vivo simeprevir, exhibited the greatest synergy with a synergy volume of 97.2 μM2%. AL-335 also exhibited synergistic interactions with the investigational HCV NS5A inhibitor, daclatasvir, (38.0 μM2%) and the investiga-tional non-nucleoside polymerase inhibitor, setrobuvir, (29.5 μM2%) whereas the interaction with ribavirin

was additive (6.8 μM2%). Conclusions: Future IFN-free therapy for CHC will require a combination of compounds with different mechanisms of action. AL-335 demonstrates an in vitro antiviral profile that suggests it may become an important component of IFN-free combination therapy. To this end, AL-335 is currently advancing towards human clinical trials for CHC. Disclosures: Kenneth Shaw – Employment: buy Compound Library Alios Biopharma Guangyi Wang – Employment: Alios Biopharma, Inc. David B. Smith – Employment: Alios BioPharma Lawrence M. Blatt – Management Position: Alios BioPharma Julian A. Symons – Employment: Alios BioPharma

The following people have nothing to disclose: Hua Tan, Natalia Dyatkina, Leo Beigelman Background: MicroRNA-122 (miR-122) is an important host factor for hepatitis C virus (HCV). Binding of miR-122 to HCV protects the HCV genome from degradation and prevents induction of an innate immune response against the virus. Miravirsen targets miR-122 and resulted in a dose dependent and prolonged decrease of HCV RNA levels. The aim of this study was to quantify plasma levels of miR-122 at baseline and during miravirsen treatment in HCV patients. Methods: We included 16

out of 36 chronic hepatitis C (genotype 1) patients who received five injections of either 3 mg/kg (n=4), 上海皓元 5 mg/ kg (n=4), 7 mg/kg (n=4) miravirsen or placebo (n=4) over a 4 week-period in a prior phase 2a study, from whom blood samples were collected at baseline, week 1, week 4, week 6 and week 10/12. RNA was isolated from plasma with the miRCURY RNA isolation kit (Exiqon) and cDNA was synthesized using qScript microRNA cDNA Synthesis Kit (Quanta). The expression levels of miR-122 were measured by quantitative PCR and normalized for levels of miR-93 and miR-191. Results: The median plasma level of miR-122 at baseline in patients receiving miravirsen was 4.3×10^3 compared to 2.2 ×10^3 copies/nl in placebo treated patients (p=ns). During anti-miR treatment, miR-122 levels showed an average 3.7fold reduction (log 2 copies/^l) between baseline and week 1 (p=0.04), 4.5-fold reduction at week 4 (p=0.007), 7-fold reduction at week 6 (p=0.016) and 6-fold reduction at week 10/12 (p=0.006) (Figure 1).

For approved compounds, combination of AL-335 with the NS3/4A protease inhibitor, learn more simeprevir, exhibited the greatest synergy with a synergy volume of 97.2 μM2%. AL-335 also exhibited synergistic interactions with the investigational HCV NS5A inhibitor, daclatasvir, (38.0 μM2%) and the investiga-tional non-nucleoside polymerase inhibitor, setrobuvir, (29.5 μM2%) whereas the interaction with ribavirin

was additive (6.8 μM2%). Conclusions: Future IFN-free therapy for CHC will require a combination of compounds with different mechanisms of action. AL-335 demonstrates an in vitro antiviral profile that suggests it may become an important component of IFN-free combination therapy. To this end, AL-335 is currently advancing towards human clinical trials for CHC. Disclosures: Kenneth Shaw – Employment: Androgen Receptor antagonist Alios Biopharma Guangyi Wang – Employment: Alios Biopharma, Inc. David B. Smith – Employment: Alios BioPharma Lawrence M. Blatt – Management Position: Alios BioPharma Julian A. Symons – Employment: Alios BioPharma

The following people have nothing to disclose: Hua Tan, Natalia Dyatkina, Leo Beigelman Background: MicroRNA-122 (miR-122) is an important host factor for hepatitis C virus (HCV). Binding of miR-122 to HCV protects the HCV genome from degradation and prevents induction of an innate immune response against the virus. Miravirsen targets miR-122 and resulted in a dose dependent and prolonged decrease of HCV RNA levels. The aim of this study was to quantify plasma levels of miR-122 at baseline and during miravirsen treatment in HCV patients. Methods: We included 16

out of 36 chronic hepatitis C (genotype 1) patients who received five injections of either 3 mg/kg (n=4), 上海皓元 5 mg/ kg (n=4), 7 mg/kg (n=4) miravirsen or placebo (n=4) over a 4 week-period in a prior phase 2a study, from whom blood samples were collected at baseline, week 1, week 4, week 6 and week 10/12. RNA was isolated from plasma with the miRCURY RNA isolation kit (Exiqon) and cDNA was synthesized using qScript microRNA cDNA Synthesis Kit (Quanta). The expression levels of miR-122 were measured by quantitative PCR and normalized for levels of miR-93 and miR-191. Results: The median plasma level of miR-122 at baseline in patients receiving miravirsen was 4.3×10^3 compared to 2.2 ×10^3 copies/nl in placebo treated patients (p=ns). During anti-miR treatment, miR-122 levels showed an average 3.7fold reduction (log 2 copies/^l) between baseline and week 1 (p=0.04), 4.5-fold reduction at week 4 (p=0.007), 7-fold reduction at week 6 (p=0.016) and 6-fold reduction at week 10/12 (p=0.006) (Figure 1).

For approved compounds, combination of AL-335 with the NS3/4A protease inhibitor, Everolimus ic50 simeprevir, exhibited the greatest synergy with a synergy volume of 97.2 μM2%. AL-335 also exhibited synergistic interactions with the investigational HCV NS5A inhibitor, daclatasvir, (38.0 μM2%) and the investiga-tional non-nucleoside polymerase inhibitor, setrobuvir, (29.5 μM2%) whereas the interaction with ribavirin

was additive (6.8 μM2%). Conclusions: Future IFN-free therapy for CHC will require a combination of compounds with different mechanisms of action. AL-335 demonstrates an in vitro antiviral profile that suggests it may become an important component of IFN-free combination therapy. To this end, AL-335 is currently advancing towards human clinical trials for CHC. Disclosures: Kenneth Shaw – Employment: Torin 1 cost Alios Biopharma Guangyi Wang – Employment: Alios Biopharma, Inc. David B. Smith – Employment: Alios BioPharma Lawrence M. Blatt – Management Position: Alios BioPharma Julian A. Symons – Employment: Alios BioPharma

The following people have nothing to disclose: Hua Tan, Natalia Dyatkina, Leo Beigelman Background: MicroRNA-122 (miR-122) is an important host factor for hepatitis C virus (HCV). Binding of miR-122 to HCV protects the HCV genome from degradation and prevents induction of an innate immune response against the virus. Miravirsen targets miR-122 and resulted in a dose dependent and prolonged decrease of HCV RNA levels. The aim of this study was to quantify plasma levels of miR-122 at baseline and during miravirsen treatment in HCV patients. Methods: We included 16

out of 36 chronic hepatitis C (genotype 1) patients who received five injections of either 3 mg/kg (n=4), MCE公司 5 mg/ kg (n=4), 7 mg/kg (n=4) miravirsen or placebo (n=4) over a 4 week-period in a prior phase 2a study, from whom blood samples were collected at baseline, week 1, week 4, week 6 and week 10/12. RNA was isolated from plasma with the miRCURY RNA isolation kit (Exiqon) and cDNA was synthesized using qScript microRNA cDNA Synthesis Kit (Quanta). The expression levels of miR-122 were measured by quantitative PCR and normalized for levels of miR-93 and miR-191. Results: The median plasma level of miR-122 at baseline in patients receiving miravirsen was 4.3×10^3 compared to 2.2 ×10^3 copies/nl in placebo treated patients (p=ns). During anti-miR treatment, miR-122 levels showed an average 3.7fold reduction (log 2 copies/^l) between baseline and week 1 (p=0.04), 4.5-fold reduction at week 4 (p=0.007), 7-fold reduction at week 6 (p=0.016) and 6-fold reduction at week 10/12 (p=0.006) (Figure 1).

70 A 67Ga radionuclide

scan may show uptake in the right lower quadrant, although there may also be activity in the gastrointestinal tract, the liver, and the spleen. However, both tumor and inflammatory cells can bind gallium.71,72 The use of indium-labelled white cells will avoid this problem and will not be affected by neutropenia. Arteriography www.selleckchem.com/products/3-methyladenine.html may show increased cecal vascularity with mucosal staining and dilatation of branches of the superior mesenteric artery. Also, there may be arteriovenous shunting as suggested by dilated, early-filling veins.73 Embolization of bleeding sites can be done although there is always a risk of transmural necrosis and perforation.35,73 Colonoscopy is not usually performed due to the severe leukopenia, thrombocytopenia, and fragile bowel wall but may reveal nodularity, ulceration, and hemorrhage.50 Macroscopically, the affected bowel is edematous, hemorrhagic, and thickened, with diffuse ischemic colitis in 69% of cases, a finding associated with increased mortality.36 The management of NE is controversial as it is based on only small case studies.36 Initially, patients should receive bowel rest, fluids, antibiotics, and, if needed, recombinant granulocyte colony stimulating factor. Non-operative treatment, found to be effective in many cases, is associated

with a 67% recurrence rate in one study from 1989.54 Surgery is indicated for persistent GI bleeding, perforation, uncontrolled sepsis, and an intra-abdominal process normally requiring surgery. Localized peritoneal Venetoclax concentration signs do not constitute an adequate reason for intervention.35 For necrosis or perforation, a right hemicolectomy is advised.39 A cecostomy and drainage may be adequate in some cases.74 In cases where surgery is done on an emergency basis with an unprepared bowel or with perforation or gross peritonitis, a two-stage procedure rather than a primary anastomosis is advisable.75 It is controversial as to whether to resect a bowel that is thickened and edematous without perforation or necrosis.53 Complications developed

in 4.6% of patients with leukemia, most commonly abscess formation, such as hepatic abscess probably from seeding from the portal circulation;35 intussusception;29 postinflammatory MCE公司 colonic stricture;76 and obstruction.28 Acute abdominal conditions can be seen in both acute and chronic leukemias (5.3% and 2.6%, respectively).31 In acute leukemia, these episodes usually occur during periods of chemotherapy and are related to the primary disease, such as neutropenic colitis or splenic rupture. Acute cholecystitis may be managed with antibiotics delaying surgery until recovery of hematopoiesis.48 Rarely, myeloid leukemia cells infiltrate the gallbladder resulting in cholecystitis.27 Fatal acute abdominal catastrophes may occur with ischemic bowel.77 In chronic leukemia, abdominal conditions tend to develop randomly during the course of disease and are similar to those seen in an elderly population.

70 A 67Ga radionuclide

scan may show uptake in the right lower quadrant, although there may also be activity in the gastrointestinal tract, the liver, and the spleen. However, both tumor and inflammatory cells can bind gallium.71,72 The use of indium-labelled white cells will avoid this problem and will not be affected by neutropenia. Arteriography Temozolomide may show increased cecal vascularity with mucosal staining and dilatation of branches of the superior mesenteric artery. Also, there may be arteriovenous shunting as suggested by dilated, early-filling veins.73 Embolization of bleeding sites can be done although there is always a risk of transmural necrosis and perforation.35,73 Colonoscopy is not usually performed due to the severe leukopenia, thrombocytopenia, and fragile bowel wall but may reveal nodularity, ulceration, and hemorrhage.50 Macroscopically, the affected bowel is edematous, hemorrhagic, and thickened, with diffuse ischemic colitis in 69% of cases, a finding associated with increased mortality.36 The management of NE is controversial as it is based on only small case studies.36 Initially, patients should receive bowel rest, fluids, antibiotics, and, if needed, recombinant granulocyte colony stimulating factor. Non-operative treatment, found to be effective in many cases, is associated

with a 67% recurrence rate in one study from 1989.54 Surgery is indicated for persistent GI bleeding, perforation, uncontrolled sepsis, and an intra-abdominal process normally requiring surgery. Localized peritoneal Palbociclib research buy signs do not constitute an adequate reason for intervention.35 For necrosis or perforation, a right hemicolectomy is advised.39 A cecostomy and drainage may be adequate in some cases.74 In cases where surgery is done on an emergency basis with an unprepared bowel or with perforation or gross peritonitis, a two-stage procedure rather than a primary anastomosis is advisable.75 It is controversial as to whether to resect a bowel that is thickened and edematous without perforation or necrosis.53 Complications developed

in 4.6% of patients with leukemia, most commonly abscess formation, such as hepatic abscess probably from seeding from the portal circulation;35 intussusception;29 postinflammatory MCE公司 colonic stricture;76 and obstruction.28 Acute abdominal conditions can be seen in both acute and chronic leukemias (5.3% and 2.6%, respectively).31 In acute leukemia, these episodes usually occur during periods of chemotherapy and are related to the primary disease, such as neutropenic colitis or splenic rupture. Acute cholecystitis may be managed with antibiotics delaying surgery until recovery of hematopoiesis.48 Rarely, myeloid leukemia cells infiltrate the gallbladder resulting in cholecystitis.27 Fatal acute abdominal catastrophes may occur with ischemic bowel.77 In chronic leukemia, abdominal conditions tend to develop randomly during the course of disease and are similar to those seen in an elderly population.

pylori in this group of adolescents was high. However, there was no correlation between H. pylori and HAV infection rates. Hence, factors contributing to the transmission source and route seem to be

different. “
“This review summarizes studies on the epidemiology and public health implications of Helicobacter pylori published in peer-reviewed journals from April 2010 through March 2011. Prevalence rates vary widely between different geographical regions and ethnic groups. An interesting study from the USA identified the degree of African ancestry as an independent predictor of H. pylori infection. Two studies have demonstrated BGB324 concentration early childhood as the period of transmission of infection and identified an infected sibling as an important risk factor. An oral–oral route of spread has been substantiated with several studies showing the presence of H. pylori in the oral cavity. Studies have shown the presence of H. pylori in drinking water and the role of poor living conditions and sanitation in H. pylori infection, Selleckchem BVD-523 supporting an oral–fecal route of spread. Screening for H. pylori as a gastric cancer pre-screening strategy has been described in Japan, and the importance of H. pylori eradication as a gastric cancer–prevention strategy has now

been further emphasized in Japanese guidelines. Two studies have shown a decrease in the burden of dyspepsia and peptic ulcer disease with H. pylori eradication. This article presents a review of the literature concerning the epidemiology and public health implications of Helicobacter pylori infection published from April 2010 till March 2011. The authors searched PubMed and Embase using MeSH terms “Helicobacter infections/epidemiology” and “Helicobacter infections/prevention and control”, and repeated the PubMed search independently using MeSH term “Helicobacter” alone

and using the set operator AND with the terms “Epidemiology”, MCE公司 “Prevalence”, “Incidence”, “Transmission”, “Risk Factors”, “Prevention and Control” or “Environment”. The identified literature is summarized below by subtopic: prevalence; incidence; transmission; risk factors; and public health implications. Serology was the most common method of diagnosis used in these studies, but several studies were endoscopy-based and diagnoses were then made by rapid urease test (RUT), culture, immuno-histochemistry or histology. A few studies utilized the urea breath test (UBT) or the stool antigen test (SAT). In an interesting and important study, Gong et al. [1] compared the accuracy of serological testing to histological diagnosis in a gastric cancer screening field survey in northern China. They found that serological testing using a commercially available kit and utilizing the recommended cut-off level underestimated the prevalence of H. pylori by almost 30% and emphasized the need for local validation of serological tests. We have grouped the studies according to the different geographical regions for ease of reference (Table 1).

Furthermore, the tumors, as well as surrounding tissues in HNF4α-KO mice showed extensive up-regulation of c-Myc and Cyclin D1. These data further support the hypothesis http://www.selleckchem.com/products/R788(Fostamatinib-disodium).html that HNF4α inhibits hepatocyte proliferation by inhibiting the c-Myc gene network. RNA-seq analysis revealed several up-regulated genes, which are potentially negatively regulated

by HNF4α. A few of these genes have a putative HNF4α binding site on their promoter and may be targets of direct inhibition by HNF4α (Ect2 and Cdc20), one of which we have confirmed in previous studies using ChIP (Ect2)19; however, a vast number of the up-regulated genes do not have an HNF4α binding site, including Cyclin D1 and c-Myc, and direct regulation of these genes at the level of transcription is unlikely. It is possible that HNF4α may regulate these genes indirectly by way of an intermediary pathway, or by way of microRNAs (miRNAs), as shown by Hatziapostolou et al.28 They provide

evidence of an “HNF4α circuit” involving miR-124, IL6R, STAT3, and miR-24/miR-629 in the regulation of hepatocarcinogenesis. They show a correlation between the down-regulation of HNF4α and miR-24 and an up-regulation of IL6R and STAT3 associated with the progression of HCC. We cannot comment Z-VAD-FMK mw on the expression of miRs in our model at this time, but we do not observe an increase in IL6R or STAT3. This may be due to a lack of inflammatory responses within our model, which may be a mediating event in the activation of the “HNF4α circuit.” With this said,

it is still very much a possibility that HNF4α is regulating MCE公司 many of the gene expression changes that we observe by an indirect mechanism involving miRNAs. Taken together, our data indicate that HNF4α is not only an important factor in the regulation of hepatocyte differentiation, but also critical for inhibition of hepatic proliferation. Our study sheds light on the mechanism of HNF4α-mediated inhibition of cell proliferation and indicates that HNF4α inhibits hepatocyte proliferation by down-regulation of promitogenic genes such as c-Myc. These data suggest a novel role as a tumor suppressor and highlight HNF4α as a potential therapeutic target, as well as a prognostic marker, for liver cancers. Additional Supporting Information may be found in the online version of this article. “
“Background. Sympathetic nervous system (SNS) activation of ascitic crrhosis reduces fluid delivery to the Henle’s loop and makes responses to diuretics negligible. Sympatholytic α2-ad-renoceptor agonists, associated with diuretics, may therefore improve natriuresis in advanced cirrhosis. Paradoxically, also α1-adrenergic agonists may improve systemic hemodynamics and sodium excretion in advanced cirrhosis. Aims & Methods.

BA standards were purchased either from Steraloids, Inc. (Newport, RI) or from Sigma-Aldrich (St. Louis, MO). All other chemicals were purchased from Sigma-Aldrich unless noted otherwise. Seven-week-old male C57BL/6 wild-type (WT) mice were purchased from Charles River Laboratories, Inc. (Wilmington, MA). Oatp1b2-null mice were engineered in our laboratory.6 Oatp1b2-null mice were back-crossed into a C57BL/6 background, and >99% congenicity was confirmed by the speed congenics group at Jackson Laboratories (Bar Harbor, ME). WT mice were acclimated for

at least 1 week in an American Animal Associations Laboratory IWR-1 in vitro Animal Care accredited facility under a standard temperature-, light-, and humidity-controlled environment. Mice had free access to Laboratory Rodent Chow 8604 (Harlan, Madison, WI) and drinking water. Studies were approved by the University of Kansas Medical Center Institutional Animal Care and Use Committee. Blood was collected from the suborbital veins of 8-week-old WT and Oatp1b2-null mice anesthetized with 50 mg/kg pentobarbitol (n = 6), and the livers and ilea were removed. Serum samples were separated using buy NVP-LDE225 Microtainer separating tubes (BD Biosciences,

San Jose, CA). Liver and ileum samples were frozen in liquid nitrogen and stored at −80°C until further analysis. Separate groups of 8-week-old WT and Oatp1b2-null mice (n = 6) were anesthetized MCE公司 intraperitoneally

with a ketamine/midazolam mixture (100 and 5 mg/kg, respectively), and the common bile duct of each mouse was cannulated through a high abdominal incision with the shaft of a 30-gauge needle attached to PE-10 tubing. After collection of 10-minute pre-bile, bile samples were collected for two 15-minute periods in preweighed 0.6-mL microcentrifuge tubes on ice. The volumes of bile were determined gravimetrically, using 1.0 for specific gravity. The plasma elimination of BAs was performed on 28- to 38-g, 9- to 12-month-old, age-matched Oatp1b2-null and WT mice (n = 6). The mice were anesthetized intraperitoneally with ketamine/midazolam (100 and 5 mg/kg, respectively), and the body temperature of each mouse was maintained at 37°C with a heating pad. Subsequently, the right carotid artery was cannulated with PE-10 tubing. To avoid possible renal elimination of BAs, the renal pedicles were ligated through a median abdominal incision. To prevent the enterohepatic recirculation of BAs, the common bile duct was cannulated as described above.