Hepatic Bile Duct Research Articles

Murine autoimmune cholangitis requires two hits: cytotoxic KLRG1+ CD8 effector cells and defective T regulatory cells (BA13P.122)

Abstract Primary biliary cirrhosis (PBC) is an enigmatic disease mediated by autoimmune destruction of cholangiocytes in hepatic bile ducts. Early immunological events in PBC are poorly understood; clinical signs of disease occur very late in the pathological process. We have used our unique murine model of PBC in dominant-negative TGF-β receptor type II transgenic mice to delineate critical early immunopathological pathways, and previously showed that dnTGFβRII CD8 T cells alone transfer biliary disease. Herein we report significantly increased numbers of hepatic terminally differentiated (KLRG1+) CD8 T cells in dnTGFβRII compared to B6; these cells are the major cytotoxic CD8 subset. Unexpectedly, the disease is not simply CD8 T cell intrinsic since mixed bone marrow chimeric mice were protected from biliary disease. Adoptive transfer studies showed that co-transfer of B6 but not dnTGFβRII Tregs prevented dnTGFβRII CD8 T cell mediated cholangitis; the protection was associated with decreased numbers of hepatic KLRG1+ CD8 T cells and with elimination of hepatic dnTGFβRII CD8 mediated cholangiocyte cytotoxicity. DnTGFβRII Treg cells were inferior in suppressing effector CD8 T cells in vitro compared to wild type B6 Tregs. In conclusion, these results emphasize that autoimmune cholangitis requires defects in both the T effector and regulatory compartments, which has important implications for understanding the early pathogenesis of human PBC.

Murine autoimmune cholangitis requires two hits: Cytotoxic KLRG1+ CD8 effector cells and defective T regulatory cells

Primary biliary cirrhosis (PBC) is an enigmatic disease mediated by autoimmune destruction of cholangiocytes in hepatic bile ducts. The early immunological events leading to PBC are poorly understood; clinical signs of disease occur very late in the pathological process. We have used our unique murine model of PBC in dominant-negative TGF-β receptor type II transgenic mice to delineate critical early immunopathological pathways, and previously showed that dnTGFβRII CD8 T cells transfer biliary disease. Herein we report significantly increased numbers of hepatic dnTGFβRII terminally differentiated (KLRG1+) CD8 T cells, a CD8 subset previously shown to be enriched in antigen specific cells during hepatic immune response to viral infections. We performed bone marrow chimera studies to assess whether dnTGFβRII CD8 mediated disease was cell intrinsic or extrinsic. Unexpectedly, mixed (dnTGFβRII and B6) bone marrow chimeric (BMC) mice were protected from biliary disease compared to dnTGFβRII single bone marrow chimerics. To define the protective B6 cell subset, we performed adoptive transfer studies, which showed that co-transfer of B6 Tregs prevented dnTGFβRII CD8 T cell mediated cholangitis. Treg mediated disease protection was associated with significantly decreased numbers of hepatic KLRG1+ CD8 T cells. In contrast, co-transfer of dnTGFβRII Tregs offered no protection, and dnTGFβRII Treg cells were functionally defective in suppressing effector CD8 T cells in vitro compared to wild type B6 Tregs. In vitro cholangiocyte cytotoxicity assays demonstrated significantly increased numbers of cytotoxic hepatic dnTGFβRII KLRG1+ CD8 cells compared to B6. Protection from disease by B6 Tregs was associated with elimination of hepatic dnTGFβRII CD8 mediated cholangiocyte cytotoxicity. These results emphasize that autoimmune cholangitis requires defects in both the T effector and regulatory compartments, and that an intrinsic T cell effector defect is not sufficient to mediate autoimmune biliary disease in the setting of intact immune regulation. These results have important implications for understanding the early pathogenesis of human PBC.

Intraductal papillary neoplasm of bile duct in liver with chronic hepatitis B and hepatocellular carcinoma: case report

Intraductal papillary neoplasms of the bile duct (IPNB) are a subtype of bile duct tumor, characterized by intraductal growth of neoplastic cholangiocytes with delicate fibrovascular cores. IPNB arise from multipotent stem cells which are located at the peribiliary glands in the ductal wall of large bile ducts. The differences in the histogenesis of IPNB and hepatocellular carcinoma (HCC) make the coexistence of both tumors rare. We herein report a 60-year-old Thai man with IPNB (diffuse involving the right hepatic duct and posterior segment bile ducts) and a 2.5 × 15 cm HCC in the subcapsular region of S7 in the liver, with chronic hepatitis B. Both IPNB and HCC are inflammation-related tumors. IPNB is associated with chronic biliary inflammation. Both HBV and HCV are risks factors for HCC and cholangiocarcinoma. Concurrent cases of HCC with precursor lesions of IPBN, and invasive IPNB in chronic hepatitis B and hepatitis B with cirrhosis, respectively, have been reported. Our case provides further support that HBV is a risk factor for bile duct cancers.

Pathological changes in primary cilia: A novel mechanism of graft cholangiopathy caused by prolonged cold preservation in a rat model of orthotopic liver transplantation.

To study the impairment of cholangiocyte primary cilia caused by prolonged cold preservation and its correlation with graft cholangiopathy after orthotopic liver transplantation (OLT). Subjects were 60 male Wistar rats that were divided into 2 groups: a control group (n = 30) receiving a donor liver preserved for 1 h and a study group (n = 30) receiving a donor liver preserved for 12 h. A two-cuff method was used to establish the OLT model, and the hepatic artery and bile ducts were reconstructed using stents. Samples were collected 2, 8, and 16 weeks after surgery, and 5 samples were collected from each group at each time point. Serum biochemical indicators were measured, morphological changes in intrahepatic bile ducts and cholangiocyte primary cilia were observed using an optical microscope and scanning electronic microscope, respectively, and the ciliary marker (α-tubulin) and membrane proteins (PC-1, TRPV4, and P2Y12) were detected using immunofluorescence analysis and Western blotting. In the study group, phlogocytes infiltrated around bile ducts and bile ducts proliferated markedly at 8 weeks. At 16 weeks, the biliary structures were indistinct and some bile ducts disappeared, a large amount of collagen was deposited, numerous phlogocytes infiltrated around ducts, some biliary epithelial cells (BECs) were deformed or dead, and primary cilia disappeared. In the control group, the intrahepatic bile ducts and BECs were nearly intact and the primary cilia were present. In the study group, the expression of α-tubulin, polycystin-1 (PC-1), TRPV4, and P2Y12 in bile ducts disappeared completely after 8 weeks. In the control group, expression of the marker and proteins decreased at 2 weeks and increased slightly after 8 weeks. These results suggest that the study group had dysfunctional primary cilia at the start of OLT and that this dysfunction was irreversible. In the control group, the primary cilia defects and subsequent biliary injury were temporary. Thus, prolonged cold preservation of a donor liver may cause graft cholangiopathy by altering the integrity and functions of cholangiocyte primary cilia.

The Value of Cholangiography through Jackson-Pratt Drains in the Management of Postoperative Biliary Injuries

Iatrogenic biliary injury is the most significant complication after laparoscopic cholecystectomy. We present our experience with an alternative diagnostic approach using transcatheter cholangiography (TCC) through a Jackson-Pratt (JP) drain and discuss potential benefits and limitations of the technique. From March 2002 to February 2012, 40 patients with major postoperative biliary injury underwent biliary reconstruction at our institution. Mean age was 51.7 ± 18.1 years (range, 19 to 86 years) with 30 (75%) females. Seventeen (42.5%) injuries were detected intraoperatively and in 13 (32.5%) cases, JP drains were placed for biliary drainage. Lesions were classified according to Bismuth grade: I (10 patients [25%]), II (10 patients [25%]), III (six patients [15%]), IV (10 patients [25%]), and V (four patients [10%]). TCC was performed in seven patients with JP drains (53.8%). It fully defined the injury site in three cases of limited magnetic resonance cholangiopancreatography (MRCP) such as common hepatic duct and common bile duct leaks and in four cases (57.1%) that endoscopic retrograde cholangiopancreatography (ERCP) was limited as a result of clipping of the distal common bile duct. TCC showed promising results in cases of limited MRCP and ERCP such as fistulous orifices or leakage. It may represent an alternative adjunct in the diagnostic armamentarium of complex biliary injuries.

Biliary tract intraductal papillary mucinous neoplasm: A brief report and review of literature

Biliary Tract Intraductal Papillary Mucinous Neoplasm (BT-IPMN) is a very rare entity, gradually emerging into attention as sporadic cases are being reported worldwide. In this brief report we discuss about such an entity from our part of the world, based on a case from our institution. A 47-year-old female was referred to our department with jaundice, intermittent fever with chills and rigor of 6 weeks duration. Initial evaluation revealed obstructive jaundice with distended gall bladder. Imaging with ultrasonogram (USG) and magnetic resonance imaging (MRI) showed hugely dilated intra and extrahepatic biliary radicles with multiple and diffuse soft tissue lesions filling the common bile duct (CBD) extending to the ductal system of left lobe of liver. A side viewing endoscopy demonstrated mucin extruding from a prominent ampulla of Vater. The patient was managed successfully by left hepatectomy with pancreaticoduodenectomy (HPD). Gross pathological examination of the specimen showed marked dilatation of intra and extra hepatic bile ducts with multiple polypoidal lesions and plenty of mucin filling the entire biliary ductal system. Histopathology revealed predominantly intraductal papillary mucinous adenocarcinoma at the hilum extending to left bile duct with diffuse dysplastic changes throughout the biliary tree. Thus the clinical, radiological and pathological features of this lesion clearly fit into the diagnosis of BT-IPMN, which is slowly being established as a definite clinical entity with features much similar to its pancreatic counterpart.

Unclassified Diffuse Ductal Cholangiocarcinoma; Report of a Case

Cholangiocarcinoma (CCA) is the second most common malignant tumor of the liver. It is simply classified as intrahepatic and extrahepatic CCA (including perihilar and distal extra hepatic CCA) according to the anatomic localization. Various classification systems were described for staging cholangiocarcinoma. We represent an interesting case of cholangiocarcinoma which is in the shadow area of classification by involving intrahepatic, hilar, and distal extra hepatic bile ducts. To our knowledge, this is the first case in the literature with diffuse bile duct involvement.

Effects of tissue mechanical properties on susceptibility to histotripsy-induced tissue damage

Histotripsy is a non-invasive tissue ablation method capable of fractionating tissue by controlling acoustic cavitation. To determine the fractionation susceptibility of various tissues, we investigated histotripsy-induced damage on tissue phantoms and ex vivo tissues with different mechanical strengths. A histotripsy bubble cloud was formed at tissue phantom surfaces using 5-cycle long ultrasound pulses with peak negative pressure of 18 MPa and PRFs of 10, 100, and 1000 Hz. Results showed significantly smaller lesions were generated in tissue phantoms of higher mechanical strength. Histotripsy was also applied to 43 different ex vivo porcine tissues with a wide range of mechanical properties. Gross morphology demonstrated stronger tissues with higher ultimate stress, higher density, and lower water content were more resistant to histotripsy damage in comparison to weaker tissues. Based on these results, a self-limiting vessel-sparing treatment strategy was developed in an attempt to preserve major vessels while fractionating the surrounding target tissue. This strategy was tested in porcine liver in vivo. After treatment, major hepatic blood vessels and bile ducts remained intact within a completely fractionated liver volume. These results identify varying susceptibilities of tissues to histotripsy therapy and provide a rational basis to optimize histotripsy parameters for treatment of specific tissues.

Telmisartan Ameliorates Fibrocystic Liver Disease in an Orthologous Rat Model of Human Autosomal Recessive Polycystic Kidney Disease

Human autosomal recessive polycystic kidney disease (ARPKD) produces kidneys which are massively enlarged due to multiple cysts, hypertension, and congenital hepatic fibrosis characterized by dilated bile ducts and portal hypertension. The PCK rat is an orthologous model of human ARPKD with numerous fluid-filled cysts caused by stimulated cellular proliferation in the renal tubules and hepatic bile duct epithelia, with interstitial fibrosis developed in the liver. We previously reported that a peroxisome proliferator activated receptor (PPAR)-γ full agonist ameliorated kidney and liver disease in PCK rats. Telmisartan is an angiotensin receptor blocker (ARB) used widely as an antihypertensive drug and shows partial PPAR-γ agonist activity. It also has nephroprotective activity in diabetes and renal injury and prevents the effects of drug-induced hepatotoxicity and hepatic fibrosis. In the present study, we determined whether telmisartan ameliorates progression of polycystic kidney and fibrocystic liver disease in PCK rats. Five male and 5 female PCK and normal control (+/+) rats were orally administered 3 mg/kg telmisartan or vehicle every day from 4 to 20 weeks of age. Treatment with telmisartan decreased blood pressure in both PCK and +/+ rats. Blood levels of aspartate amino transferase, alanine amino transferase and urea nitrogen were unaffected by telmisartan treatment. There was no effect on kidney disease progression, but liver weight relative to body weight, liver cystic area, hepatic fibrosis index, expression levels of Ki67 and TGF-β, and the number of Ki67- and TGF-β-positive interstitial cells in the liver were significantly decreased in telmisartan-treated PCK rats. Therefore, telmisartan ameliorates congenital hepatic fibrosis in ARPKD, possibly through the inhibition of signaling cascades responsible for cellular proliferation and interstitial fibrosis in PCK rats. The present results support the potential therapeutic use of ARBs for the treatment of fibrocystic liver disease in ARPKD patients.

Investigation of Hepatobiliary Disposition of Doxorubicin Following Intrahepatic Delivery of Different Dosage Forms

Unresectable, intermediate stage hepatocellular carcinoma (HCC) is often treated palliatively in humans by doxorubicin (DOX). The drug is administered either as a drug-emulsified-in-Lipiodol (DLIP) or as drug loaded into drug eluting beads (DEB), and both formulations are administered intrahepatically. However, several aspects of their in vivo performance in the liver are still not well-understood. In this study, DLIP and DEB were investigated regarding the local and systemic pharmacokinetics (PK) of DOX and its primary metabolite doxorubicinol (DOXol). An advanced PK-multisampling site acute in vivo pig model was used for simultaneous sampling in the portal, hepatic, and femoral veins and the bile duct. The study had a randomized, parallel design with four treatment groups (TI-TIV). TI (n = 4) was used as control and received an intravenous (i.v.) infusion of DOX as a solution. TII and TIII were given a local injection in the hepatic artery with DLIP (n = 4) or DEB (n = 4), respectively. TIV (n = 2) received local injections of DLIP in the hepatic artery and bile duct simultaneously. All samples were analyzed for concentrations of DOX and DOXol with UPLC-MS/MS. Compared to DLIP, the systemic exposure for DOX with DEB was reduced (p < 0.05), in agreement with a slower in vivo release. The approximated intracellular bioavailability of DOX during 6 h appeared to be lower for DEB than DLIP. Following i.v. infusion (55 min), DOX had a liver extraction of 41 (28-53)%, and the fraction of the dose eliminated in bile of DOX and DOXol was 20 (15-22)% and 4.2 (3.2-5.2)%, respectively. The AUCbile/AUCVP for DOX and DOXol was 640 (580-660) and 5000 (3900-5400), respectively. In conclusion, DLIP might initially deliver a higher hepatocellular concentration of DOX than DEB as a consequence of its higher in vivo release rate. Thus, DLIP delivery results in higher intracellular peak concentrations that might correlate with better anticancer effects, but also higher systemic drug exposure and safety issues.

Histopathological evaluation of resected colorectal cancer liver metastases: what should be done?

Histological reporting of hepatic resections of colorectal liver metastases (CRLMs) is limited to confirmation of diagnosis and evaluation of resection margins. More exhaustive diagnostic reporting might be warranted. Here, we critically and systematically review the potentially important histological prognostic factors in CRLM. Histological features such as intrahepatic spread, resection margins, and tumour response to neoadjuvant chemotherapy have been defined. Intrahepatic spread (venous, lymphatic, bile duct and perineural invasion) was evaluated in a number of studies. Meta-analysis demonstrated a clear correlation between 5-year overall survival and both portal vein invasion (RR 1.8, 95% CI 1.3-2.5) and lymphatic invasion (RR 1.7, 95% CI 1.4-2.0). The impact of hepatic vein invasion and bile duct invasion on outcome is not clear. Perineural invasion was linked to survival in one study. Resection margin is an important prognostic factor; however, the significance of the width of negative margins remains controversial. Various studies have evaluated tumour response to neoadjuvant chemotherapy, but different grading systems were used, and definite recommendations cannot be made. In conclusion, with the high incidence of CRLM and the increase in the number of hepatic resections, we need well-defined prognostic factors, studied in homogeneous patient populations, to optimize diagnostic work-up. This review identifies several of these factors.

One-stop shop for 3-dimensional anatomy of hepatic vasculature and bile duct with special reference to biliary image reconstruction.

Three-dimensional CT has become an essential tool for successful hepatic surgery. Up to now, efforts have been made to simultaneously visualize hepatic vasculature and bile ducts. Herein, we introduce a new one-stop shop approach to hepatic 3D-anatomy, using a standard enhanced MDCT alone. A 3D-reconstruction of hepatic vasculature was made using data from contrast enhanced MDCT and SYNAPSE VINCENT software. We identified bile ducts from axial 2D image, and then reconstructed the 3D image. Both hepatic vasculature and bile duct images were integrated into a single image and it was compared with the 3D image, utilized with MRCP or DIC-CT. The first branches of both the right and left hepatic ducts were hand-traced and visualized for all 100 cases. The second branches of these ducts were visualized in 69 cases, and only the right second branch was recognized in 52 cases. Anomalous variations of bile ducts, such as posterior branch joining into common hepatic duct, were recognized in 12 cases. These biliary tract variations were all confirmed by MRCP or DIC-CT. Our new one-stop shop approach using the 3D imaging technique might contribute to successful hepatectomy as well as reduce medical costs and radiation exposure by omission of MRCP and DIC-CT.

Diagnosis and surgical treatment of aberrant right posterior hepatic bile duct injury: a report of 28 patients

################ Key words: Bile ducts; Variation; Wounds and injures; Diagnosis; Therapy

Abstract 329: HSF-1 Knockout Enhances Dietary Cholesterol Metabolism by Inducing CYP7A1 Gene Expression and Attenuates Atherosclerosis

Objective: Coronary heart disease and diabetes are highly prevalent among obese populations due to aberrant dietary cholesterol metabolism. Here we investigated the effect of heat shock factor-1 (HSF-1) on atherosclerosis and dietary cholesterol metabolism. Methods and Results: Atherogenic western diet-induced weight gain was reduced in HSF-1 and LDLr double knock out mice (HSF-1 -/- /LDLr -/- ), compared to LDLr -/- mice. Atherosclerotic lesion growth in aortic arch and carotid regions was retarded. Also, repression of PPAR-γ2 and AMPKα expression in adipose tissue, low hepatic steatosis, and lessened plasma adiponectins and lipoproteins were observed. Furthermore, reduced heat shock proteins and their mRNA levels in atherosclerotic lesions correlated with reduction in lesion burden. In HSF-1 -/- /LDLr -/- liver, higher cholesterol 7α hydroxylase (CYP7A1, the rate limiting enzyme in the synthesis of bile acid from cholesterol) and MDR1/p-glycoprotein (bile salt transporter across the hepatocyte canalicular membrane) gene expressions were observed, consistent with higher bile acid sequestration and larger hepatic bile ducts. HSF-1 deletion, however, upregulated both CYP7A1 enzyme and MDR1/p-glycoportein expression and activities, due to removal of its repressive binding in the CYP7A1 and MDR1 gene promoters. This increased the conversion of cholesterol into 7-α-hydroxycholesterol and bile acid, and dietary cholesterol metabolism. Conclusions: HSF-1 ablation not only eliminates heat shock response to retard atherosclerosis, but it also transcriptionally upregulates CYP7A1 and MDR1/P-gp axis to increase cholesterol metabolism. Therefore, HSF-1 is a metabolic regulator of dietary cholesterol and a major contributor to heart disease among obese population.

Does a combination of opisthorchiasis and ethyl alcohol consumption enhance early cholangiofibrosis, the risk of cholangiocarcinoma?

Combination of Opisthorchis viverrini infection and other factors could drive cholangiocarcinoma (CCA) development in Southeast Asia. However, other CCA factors are obscure. Alcohol consumption is well known in the risk for several cancers, but there is no report in CCA development. Therefore, the present study was to clarify whether drinking alcohol increases the liver pathology of Opisthorchis viverrini (OV) infection which may be the CCA risk. Experimental Syrian hamsters were divided into two groups: (1) infected with OV alone (OV); and (2) infected with OV plus administration of drinking alcohol (OV + ALC) for various lengths of time, i.e., 1, 2, 3, and 6 months. Hamster livers were collected for analysis of histopathological changes through hematoxylin and eosin, Sirius red, and immunohistostaining for proliferating cell nuclear antigen (PCNA) and cytokeratin 19 (CK19). Syrian hamster sera were used for liver function tests. Observed histopathological changes consisted primarily of aggregations of inflammatory cells surrounding the hepatic bile duct, especially at the hilar region, in both OV and OV + ALC groups; however, there was a difference in virulence. The OV + ALC group showed greater severity than the OV group. Moreover, in addition to aggregations of inflammatory cells, new bile duct formation (including hepatic cell death) was observed in subcapsular hepatic tissue. Bile duct proliferation, as determined by positive immunohistochemical staining for CK19 and PCNA, was correlated with the histopathology. Increased fibrosis was observed in subcapsular liver tissue. The present study suggests that alcohol consumption can exacerbate cholangiofibrosis, cholangitis, and lithiasis, which are risk factors for CCA.

Histopathological evaluation of resected colorectal cancer liver metastases: what should be done?

Histological reporting of hepatic resections of colorectal liver metastases (CRLMs) is limited to confirmation of diagnosis and evaluation of resection margins. More exhaustive diagnostic reporting might be warranted. Here, we critically and systematically review the potentially important histological prognostic factors in CRLM. Histological features such as intrahepatic spread, resection margins, and tumour response to neoadjuvant chemotherapy have been defined. Intrahepatic spread (venous, lymphatic, bile duct and perineural invasion) was evaluated in a number of studies. Meta-analysis demonstrated a clear correlation between 5-year overall survival and both portal vein invasion (RR 1.8, 95% CI 1.3-2.5) and lymphatic invasion (RR 1.7, 95% CI 1.4-2.0). The impact of hepatic vein invasion and bile duct invasion on outcome is not clear. Perineural invasion was linked to survival in one study. Resection margin is an important prognostic factor; however, the significance of the width of negative margins remains controversial. Various studies have evaluated tumour response to neoadjuvant chemotherapy, but different grading systems were used, and definite recommendations cannot be made. In conclusion, with the high incidence of CRLM and the increase in the number of hepatic resections, we need well-defined prognostic factors, studied in homogeneous patient populations, to optimize diagnostic work-up. This review identifies several of these factors.

Differential Activation of Diverse Glutathione Transferases of Clonorchis sinensis in Response to the Host Bile and Oxidative Stressors

Background Clonorchis sinensis causes chronic cumulative infections in the human hepatobiliary tract and is intimately associated with cholangiocarcinoma. Approximately 35 million people are infected and 600 million people are at risk of infections worldwide. C. sinensis excretory-secretory products (ESP) constitute the first-line effector system affecting the host-parasite interrelationship by interacting with bile fluids and ductal epithelium. However, the secretory behavior of C. sinensis in an environment close to natural host conditions is unclear. C. sinensis differs from Fasciola hepatica in migration to, and maturation in, the hepatic bile duct, implying that protein profile of the ESP of these two trematodes might be different from each other.Methodology/Principal FindingsWe conducted systemic approaches to analyze the C. sinensis ESP proteome and the biological reactivity of C. sinensis glutathione transferases (GSTs), such as global expression patterns and induction profiles under oxidative stress and host bile. When we observed ex host excretion behavior of C. sinensis in the presence of 10% host bile, the global proteome pattern was not significantly altered, but the amount of secretory proteins was increased by approximately 3.5-fold. Bioactive molecules secreted by C. sinensis revealed universal/unique features in relation to its intraluminal hydrophobic residing niche. A total of 38 protein spots identified abundantly included enzymes involved in glucose metabolism (11 spots, 28.9%) and diverse-classes of glutathione transferases (GSTs; 10 spots, 26.3%). Cathepsin L/F (four spots, 10.5%) and transporter molecules (three spots, 7.9%) were also recognized. The universal secretory proteins found in other parasites, such as several enzymes involved in glucose metabolism and oxygen transporters, were commonly detected. C. sinensis secreted less cysteine proteases and fatty acid binding proteins compared to other tissue-invading or intravascular trematodes. Interestingly, secretion of a 28 kDa σ-class GST (Cs28σGST3) was significantly affected by the host bile, involving reduced secretion of the 28 kDa species and augmented secretion of Cs28σGST3-related high-molecular-weight 85 kDa protein. Oxidative stressors induced upregulated secretion of 28 kDa Cs28σGST3, but not an 85 kDa species. A secretory 26 kDa μ-class GST (Cs26μGST2) was increased upon treatment with oxidative stressors and bile juice, while another 28 kDa σ-class GST (Cs28σGST1) showed negligible responses.Conclusions/SignificanceOur results represent the first analysis of the genuine nature of the C. sinensis ESP proteome in the presence of host bile mimicking the natural host environments. The behavioral patterns of migration and maturation of C. sinensis in the bile ducts might contribute to the secretion of copious amounts of diverse GSTs, but a smaller quantity and fewer kinds of cysteine proteases. The Cs28σGST1 and its paralog(s) detoxify endogenous oxidative molecules, while Cs28σGST3 and Cs26μGST2 conjugate xenobiotics/hydrophobic substances in the extracellular environments, which imply that diverse C. sinensis GSTs might have evolved for each of the multiple specialized functions.

The Degenerin/ENaC channel, encoded by ACCN5, serves a critical function in the vestibulocerebellum

ACCN5 encodes the BLINaC/hINaC member of the Deg/ENaC ion channel family. As we demonstrate using a novel mouse line that we engineered to contain a LacZ cassette knockin that interrupts the third exon of this gene, BLINaC has a unique expression profile for a Deg/ENaC channel. Similar to acid‐sensing channels, it is expressed in neurons. But also similar to the epithelial Na+ channel, it is expressed in epithelial cells, particularly those lining the hepatic bile duct. Thus, BLINaC is the only known mammalian Deg/ENaC channel to be expressed in both neurons and epithelial cells. In the brain, BLINaC is restrictively expressed in a subset of interneurons in the cerebellum. Specifically, BLINaC is expressed in unipolar brush cells (UBC) of lobule IX and X. These cells serve a key function in the vestibulocerebellum. Deletion of ACCN5 in our novel ACCN5 null mouse line results in abnormal coordination and balance as measured with standard tests of behavior. It is likely that this phenotype results UBC dysfunction. These results are consistent with BLINaC serving a critical function in UBC that is key to normal vestibulocerebellar function and motor control of balance.

Blood supply of the main bile duct from the caudate artery and medial subsegmental artery of the hepatic artery: Evaluation using images obtained during transcatheter arterial chemoembolization for hepatocellular carcinoma

Main bile duct necrosis develops after transcatheter arterial chemoembolization (TACE) through the caudate artery (A1) and medial subsegmental artery (A4) of the hepatic artery in the treatment of hepatocellular carcinoma. The aim of this study was to evaluate the bile duct branch (BD branch) from A1 and A4. We evaluated the origin and vascular territory of the BD branch in 11 patients who underwent selective A1 and/or A4 arteriography using arteriograms, cone-beam computed tomography (CBCT) and CT obtained 1 week after TACE. Follow-up CT and/or magnetic resonance imaging were also evaluated. The BD branch arose from the first branch (n = 4), the second branch (n = 1), and both the first and second branches (n = 1) of A1, and from the first branch of A4 (n = 5). It supplied the bilateral hepatic ducts and common hepatic duct (CHD) (n = 4), the right hepatic duct (RHD) and CHD (n = 2), RHD, CHD and common bile duct (n = 1), the left hepatic duct (LHD) and CHD (n = 2), and LHD alone (n = 2). Anastomosis between A1 or A4 and other branches was demonstrated in seven patients. Bile duct stricture developed in all nine patients 2-8 months after TACE of the BD branch and percutaneous transhepatic bile duct drainage and metallic stent placement was required in one because of jaundice. The BD branch arises from the proximal portion of A1 and A4 and mainly supplies the hepatic ducts and CHD.

Analysis of patterns of failure and prognostic factors in resected extrahepatic bile duct cancer: Implications for postoperative adjuvant radiotherapy.

266 Background: This study is aimed to analyze the patterns of failure and evaluate the prognostic factors in the patients with extrahepatic bile duct cancer (EBDC) for the potential role of postoperative adjuvant radiotherapy (PORT). Methods: We retrospectively reviewed the medical records of 106 patients with EBDC who received curative intent surgical resection. The definitions of tumor location were as follows: proximal EBDC (n = 29) from the confluent portion of the bilateral hepatic bile ducts to the junction of cystic duct, and distal EBDC (n = 77) from the junction of cystic duct to intrapancreatic portion. Nine patients underwent adjuvant chemoradiotherapy or chemotherapy. Results: The median follow-up time was 24 months for the surviving patients. Forty patients experienced locoregional failure (LRF) initially; 13 (45%) with proximal EBDC and 27 (35%) with distal EBDC. The hepatoduodenal ligament (HL) and tumor bed were the most common LRF sites. Distant metastasis (DM) occurred in 10 patients (34%) with proximal EBDC and 15 patients (19%) with distal EBDC. The liver was the most common organ of DM. In the multivariate analysis, perineural invasion (PNI) and postoperative high carbohydrate antigen (CA) 19-9 were associated with poor LRPFS. Conclusions: Both proximal and distal EBDC showed remarkable proportion of LRF. Because the HL and tumor bed are where routinely covered by PORT, it can be speculated that the addition of PORT can improve LRPFS in these patients. Especially PORT needs to be considered in patients with PNI and postoperative high CA 19-9 to improve locoregional control.