Number Of Bile Ducts Research Articles

The Impact of the Multiple Bile Ducts on Postoperative Biliary Complications In Patients Undergoing Living Donor Liver Transplantation

We aimed to investigate the correlation between bile duct number and biliary complications in patients who underwent donor liver transplantation (LDLT). We reviewed all patients who underwent LDLT in our hospital between July 2008 and December 2020. The patients were divided into 2 groups according to the number of bile ducts in the living donor graft (single duct [SD] or multiple ducts [MD]). Collected data included donor and recipient demographics, surgical data including bile duct reconstruction, and perioperative and postoperative outcomes. No prisoners were used in this study, and participants were neither coerced nor paid. The current study complies with the Helsinki Congress and the Declaration of Istanbul. All 70 patients were classified as SD (n=48) and MD (n=22). Complications related to the bile duct occurred in 27 (38.6%) patients and were more common in the MD group (54.5% vs 31.3%; odds ratio, 2.4). The MD patients had a longer operation time (1052 ± 251 vs 910 ± 215 minutes, P=.019) and a higher percentage of hepaticojejunostomy (31.8% vs 8.3%, P=.012). Donor age, graft-recipient weight ratio, cold ischemic time, and transfusion volume did not differ between groups. Twenty-one patients (77.7%) fully recovered from complications related to the bile duct, but 3 patients (4.3%) had liver graft failure. Bile duct-related complications were common in LDLT patients, despite overall good results. Multiple bile ducts may be a potent risk factor for postoperative biliary complications.

FRUCTOSE DIET INDUCES A METABOLIC REPROGRAMMING TO ENHANCE TUMOR AGGRESSIVENESS

HCC is one of the main causes of cancer-related death worldwide and has third place in mortality. One of the main risk factors is metabolic-associated fatty liver disease (MAFLD), having hepatic steatosis, and related metabolic disorders. Mexican population has the highest obesity rate in both children and adults, and the consumption of hypercaloric diets has been related to that. Also, Mexico is in the top five countries with a higher fructose-enriched diet consumption and has been proved already the relation between fructose consumption and MAFLD. Likewise, fructose has been related to metabolic rewiring in transformed cells, enhancing aggressiveness and survival. To analyze fructose role on aggressiveness promotion of HCC cells. We used C57Bl/6J mice strain (both sex) with a high Fructose diet (Fru) (33% of fructose in the drinking water, ad libitum ). Fru supplementation started with 15 days-old mice, two days after DEN was injected (10 μg/Kg, i.p), and the treatment was ended 8 months later. The UAM ethics committee approved the protocol. In vitro studies were carried out with the Huh-7 HCC cell line and we evaluated metabolic and biochemical parameters. Tissue samples were analyzed by H&E. We observed that the fructose-enriched diet group mice presented fat accumulation in the hepatocytes and also areas with a greater inflammatory infiltrate (Fru). Mice in the fructose-enriched diet + DEN (Fru/HCC) group showed a marked difference between the tumor area and the surrounding tissue and an increase in the number of bile ducts, indicating liver tissue damage. Also, we analyzed the protein content of some lipogenic enzymes and noticed an increment in fatty acid synthase (Fasn) in Fru and Fru/HCC. Due to that, we analyzed if Fru treatment was inducing metabolic rewiring in transformed cells. We obtained metabolic changes in fructose-treated cells, reducing the glycolytic pathway and the traditional Warburg effect. Then we evaluated if the Huh-7 under a Fru treatment was more dependent on mitochondria or glycolysis ATP generation. We observed a reduction in proliferation under oligomycin treatment vs . 2-DG treatment. At least, we evaluated the pentose phosphate pathway (PPP) under a Fru treatment and obtained a higher glucose-6-phosphate dehydrogenase (G-6-P DH) activity with Fru vs. only glucose (Glc). Also, G-6-P DH had a more efficient activity in the presence of Fru because the time to reach the Vmax is lower vs . Glc. Fructose induces a metabolic rewiring in cancer cells to enhance ATP production, NADPH, and nucleotides to sustain the active lipid synthesis and proliferation. The fructose-enriched diets promote an aberrant lipogenic phenotype enhancing tumor aggressiveness. Conacyt, Froteras de la Ciencia 1320. The authors declare that there is no conflict of interest.

A Novel Technique for Bile Duct Division During Laparoscopic Living Donor Hepatectomy to Overcome Biliary Complications in Liver Transplantation Recipients: "Cut and Clip" Rather Than "Clip and Cut".

This study was designed to analyze the clinical impact of a new bile duct division technique during laparoscopic living donor hepatectomy. Laparoscopic donor right hepatectomies performed by a single surgeon between December 2016 and August 2019 were included. Magnetic resonance cholangiopancreatography of the donors was reconstructed, and the length of the common channel of the dividing intrahepatic duct was measured. Patients were divided into 2 groups based on the bile duct division techniques of "clip and cut" versus "cut and clip." Outcome of bile duct division was categorized based on the graft and bile duct type and number of bile duct openings. A total of 147 transplantations were included. Outcomes in the "cut and clip" group were better than those in the "clip and cut" group with regard to obtaining an adequate division point (94.2% versus 78.8%; P = 0.019). The "cut and clip" showed superior biliary stricture-free survival than the "clip and cut" group (hazard ratio [HR], 0.456; confidence interval [CI], 0.217-0.957; P = 0.038) whereas recipients with hepaticojejunostomy (HR, 3.358; CI, 1.090-10.350; P = 0.035) and bile duct type other than type I (HR, 2.214; CI, 1.143-4.289; P = 0.018) showed increased risk for biliary stricture-free survival. Better outcome in terms of obtaining a single opening was also obtained in the "cut and clip" group (68.2% versus 84.6%; P = 0.040), and this difference was significant especially with a common channel shorter than 10 mm (45.5% versus 81.2%; P = 0.043). The "cut and clip" technique showed superior outcomes to the "clip and cut" technique in terms of fewer bile duct openings and biliary stricture-free survival of the recipients.

МОРФОЛОГИЧЕСКИЕ ИЗМЕНЕНИЯ СОСУДИСТОЙ СИСТЕМЫ ПЕЧЕНИ КРЫС ПОД ВОЗДЕЙСТВИЕМ ТИОАЦЕТАМИДА

Background. Angiogenesis plays a key role in the progression of liver fibrosis. However, the available data on morphological changes in the liver vascular system are insufficient and contradictory. Objective. The aim of the work is to study the morphological changes in the liver vascular system of rats under the influence of thioacetamide. Material and methods. Fibrosis and cirrhosis of the liver in Wistar rats were induced with thioacetamide given at a dose of 200 mg/kg of animal weight for 17 weeks. To study morphological changes, we used classical and immunohistochemical staining methods. Microscopic analysis was performed using OLYMPUS BX51 microscope and image analysis software ImageScope Color and cellSens Standard. Results. The introduction of a solution of thioacetamide through the stomach leads to a gradual increase in the progression of pathological changes. In addition, it permits to track all stages of cirrhosis development and morphological restructuring of the liver vascular system. Throughout the experiment there was intensive capillarization of the parenchyma sinusoids and neoangiogenesis in the portal tracts and connective tissue septa manifested by the formation of many venules and small veins. We also observed an increase in the area of interlobular veins, which in some places had reached gigantic proportions. Three morphological phenotypes of CD34-positive cells were revealed. In the interlobular arteries as well as interlobular, central and sublobular veins, these cells had an elongated shape and a rod-shaped dark-colored nucleus. During the transformation of liver fibrosis into cirrhosis CD34-positive cells of an elongated shape with light roundedelongated nuclei were observed in the sinusoids closer to the periphery of individual false nodules. Rounded cells with dark-colored nuclei were present in the connective tissue near the hepatic triads, in the connective tissue septa among the cells of the infiltrate and between the sharply increased number of bile ducts. Conclusions. The established complex phenotypic changes in sinusoidal endothelial cells prove a close connection between fibrogenesis and neoangiogenesis. They probably play a leading role in the development of fibrosis and restructuring of the venous system of the portal vein.

Biliary reconstruction during right lobe living donor liver transplantation: state of the problem, options and classification

Objective. To study the features and results of biliary reconstruction in right lobe living donor liver transplantation. To propose a classification of biliary reconstruction, which allows reflecting all its existing and possible variants, taking into account the anatomy of the bile ducts of the graft and recipient and technical features of reconstruction. Materials and methods. From May 2010 to June 2019 a retrospective and prospective analysis of the results of the right lobe living donor liver transplantation was performed for 256 recipients and their donors. One observation was excluded due to lack of data. Results. When the right lobe liver transplantations were performed, grafts with two bile ducts requiring biliary reconstruction most often (52%) were observed, less often grafts with one duct (32%), three ducts (12%), four ducts (3%) and in one case five ducts (<1%) were observed. With the multiple nature of the ducts in 31 cases (12% of all operations) unification ductoplasty was used. Moreover, in the group of reconstructions with the presence of two ducts and in the group with the presence of three or more ducts ductoplasty was used with equal frequency – in 18% of cases. The duct-to-duct anastomoses was formed in 157 cases (61%), bilio-entero anastomoses – in 91 cases (36%) and the combined variant – only in 7 cases (3%). In the first variant (duct-to-duct) during reconstruction in 43 cases (27% of all formed duct-to-ducts, 17% of all transplants performed), the so-called high-order bile ducts (right or left lobar, as well as cystic bile ducts) were used. This article proposes a coding and classification system for the technical features of biliary reconstruction in right lobe living donor liver transplantation. Conclusion. Right lobe living donor liver transplantation accompanied by high variability of the stage of biliary reconstruction, depending on the number of bile ducts in the graft, their condition and relative position. The analysis of our own material made it possible to propose a universal classification and coding method for the types of biliary reconstruction during right lobe living donor liver transplantation. The classification allows to reflect all existing options for biliary reconstruction, depending on the anatomy of the bile ducts of the graft and recipient, the technical features of its implementation. It also simplifies the analysis and statistical processing of the bile duct reconstruction stage in liver transplantation.

Biliary Reconstruction in Adult Living Donor Liver Transplantation: The All-Knots-Outside Technique.

Biliary complications (BCs) following living donor liver transplantation (LDLT) can lead to significant morbidity and occasional mortality. The present study describes our experience of the all‐knots‐outside technique (AKOT) of biliary reconstruction (BR) and its impact on BCs. A retrospective analysis was performed using prospectively collected data of 330 consecutive adult patients undergoing LDLT from July 2011 to February 2018 with a minimal follow‐up of 24months. Only 2.8% required hepaticojejunostomy and were excluded. In an initial 122 patients, BR was performed with the standard technique (ST), and AKOT was performed in the subsequent 208 patients. In the AKOT group, a single anastomosis was attempted even for multiple ducts whenever feasible. A major BC was defined as requiring endoscopic, percutaneous, or surgical interventions. In the AKOT group, significantly more patients received a left lobe graft (5.7% versus 18.3%; P =0.001), had shorter warm ischemia time (44.6 versus 27minutes; P <0.001), and had a left hepatic artery (LHA) in the right lobe that was used for arterial reconstruction (48 [39.3%] versus 122 [58.6%]; P =0.003). The incidence of BCs in the entire cohort was 47 (14.2%). For the ST versus AKOT groups, the overall BCs (27/122 [22.1%] versus 20/208 [9.6%]; P =0.003) and major BCs (20.5% versus 6.7%; P <0.001) decreased significantly. In the multivariate analysis, the number of bile ducts (hazard ratio [HR], 4.18; 95% confidence interval [CI], 1.62‐10.78; P =0.003), number of anastomoses (HR, 2.03; 95% CI, 1.03‐4.02; P =0.04), and technique of anastomosis (HR, 0.36; 95% CI, 0.19‐0.68; P =0.002) predicted BCs. In conclusion, in adult LDLT, with standardization of the donor and recipient surgery, preferential use of LHA for right lobe arterial reconstruction, reduction in the number of anastomoses, and AKOT for BR significantly decreased the incidence of BCs.

Idiopathic Intrahepatic Cholestasis Associated With Monoclonal Gammopathy of Undetermined Significance

Cholestasis is a common presenting symptom in various hematologic diseases. We present a patient who had non-obstructive jaundice without demonstrable malignant cell infiltration and intact intrahepatic bile ducts, which was related to monoclonal gammopathy of undetermined significance (MGUS). A 58- year- old male had history of painless jaundice, anorexia and weight lost for 2 months. He had dark urine and pale stool. The patient denied alcohol and medicine intake. Physical examination found unrecognized fever, pale conjunctiva, icteric sclera and splenic dullness on percussion. Otherwise were unremarkable. Laboratory workups were significant for hemoglobin of 8.9, a TB of 65.7, a DB of 34.1, a SGOT of 170, a SGPT of 109, a ALP of 144. Peripheral blood smear was compatible with cold-type AIHA. CT of the whole abdomen showed normal parenchyma of liver without intrahepatic duct and CBD dilatation. The spleen size was 11 cm. No intra-abdominal lymph node enlargement. Workups for liver disease including viral hepatitis (HAV, HBV, HCV, HEV), ANA, ASMA, and AMA were negative. No atypical cell was detected from bone marrow study. Serum free light chain was significantly high level for Kappa monoclonal gammopathy. Liver biopsy found mixed inflammatory cells infiltration in hepatic lobules and portal tract. There was no significant bile duct loss. No malignancy was detected. The diagnosis of idiopathic intrahepatic cholestasis associated with MGUS was made. Intravenous dexamethasone 20 mg/day was prescribed. The LFTs improved dramatically after treatment and was allowed to be discharged with prednisolone 60 mg/day. Most cases of paraneoplastic-related cholestasis are caused by cytokines-induced vanishing bile duct syndrome. However, liver biopsy results from our patient did not show significant decrease in number of bile duct. Cholestasis in our patient was likely caused by dysregulation of bile duct transporters insulted from inflammatory mediators. This phenomenon is called idiopathic intrahepatic cholestasis, which is extremely rare paraneoplastic event. To the best of our knowledge, our patient has been the first case reported in MGUS. Treatment with systemic steroids is highly effective. In conclusion, paraneoplastic syndrome presenting as idiopathic intrahepatic cholestasis is associated with MGUS. Systemic steroid is effective for the treatment of this condition.2399_A Figure 1. The hepatic lobuls showed severe cholestasis with feathery degeneration. There was mild mixed inflammatory cell infiltrate in hepatic lobules and portal tracts. Bile duct was observed without bile duct injury.2399_B Figure 2. Immunohistochemical study for CK7 demonstrated insignificant bile duct loss.2399_C Figure 3. The liver function test improved dramatically after steroids administration.

TGFβ signaling controls intrahepatic bile duct development may through regulating the Jagged1-Notch-Sox9 signaling axis.

Due to the inherent limitations of the mouse models, the molecular mechanism of TGFβ signaling involved in the development of intrahepatic bile ducts (IHBDs) has been investigated little. Here, we investigated the role of TGFβ signaling and its regulatory mechanism in IHBDs development. We demonstrate that TGFβ signaling pathway activity is essential for IHBDs development. When blocking TGFβ signaling at E10.5, the number of bile ducts in hilum was reduced more than two fold and number of CK19 positive chlangiocytes in periphery was reduced more than 3.5-fold compared with controls. We also show that alpha-smooth muscle actin (α-SMA)-immunoreactive cells are located in the portal vein mesenchyme (PVM) adjacent to the bile ducts during IHBDs development and identify the α-SMA positive cells expressing the Notch ligand Jagged1 in the periportal area. Importantly, after blocking TGFβ signaling, the expression of Jagged1 was selectively decreased in the PVM but not in biliary epithelial cells (BECs), which is associated with the transformation of portal mesenchyme cells (PMCs) into portal myofibroblasts (PMFs). In addition, Sox9, which is downstream of Notch, is decreased after blocking the TGFβ signaling pathway in the liver. Our findings uncover a novel mechanism of TGFβ signaling in controlling the development of IHBDs may through regulating the Jagged1-Notch-Sox9 signaling axis.

Clinicopathologic features of primary hepatic marginal zone lymphoma of mucosa-associated lymphoid tissue and hepatic pseudolymphoma

Objective: To study the clinicopathological features of primary hepatic extranodal marginal zone lymphoma of mucosa associated lymphoid tissue (MALT lymphoma) and hepatic pseudolymphoma, and to discuss their differential diagnosis, treatment and prognosis. Methods: Three primary hepatic MALT lymphomas and two hepatic pseudolymphomas collected from January 2012 to March 2017 in the First Affiliated Hospital of Nanjing Medical University were evaluated by HE and immunohistochemistry(IHC), in-situ hybridization and immunoglobulin (Ig) gene rearrangement detection, and the relevant literature reviewed. Results: In the three MALT lymphomas, tumor cells infiltrated the portal areas with nodular pattern, and invaded the surrounding normal liver with serpiginous configuration and formation of confluent sheets. A number of bile ducts were entrapped within the lesions, and showed lymphoepithelial lesion. Reactive lymphoid follicles were present and surrounded by tumor cells, consisting of predominantly centrocyte-like cells and monocytoid B cells. There were clusters of epithelioid histiocytes in one case. The tumor cells were positive for CD20, PAX5 and negative for CD5, CD23, CD10, bcl-6, and cyclin D1. In the two hepatic pseudolymphomas, the lesions presented as solitary nodules well-demarcated from the surrounding liver tissue; one case was partially encapsulated with fibrous tissue. Entrapped bile ducts were only found at the edge of the lesions without lymphoepithelial lesion. The lesions comprised of massive lymphoid proliferation consisting predominantly of reactive lymphoid follicles, but not monocytoid B-cells or atypical cells. By IHC, a mixture of B- and T-cell population was identified. A monoclonal rearrangement of the Ig gene was detected in all three MALT lymphomas but not in two pseudolymphomas. Interphase fluorescence in situ hybridiazation test for MALT1 break-apart gene was positive in two cases of MALT lymphomas and EBER was negative in all studied cases. Conclusions: Primary heptic MALT lymphoma and pseudolymphoma are both rare lymphoid proliferative lesions of liver. These two lesions have overlapping histological and IHC features and are top differential diagnosis to each other. A combination analysis of morphology, immunophenotype and Ig gene rearrangement is helpful to distinguish between them.

EFFECTS OF SUPPLEMENTAL FEEDING WITH SOME MEDICINAL PLANTS ON SEMEN CHARACTERISTICS, BLOOD METABOLITES AND HISTOLOGY OF SELECTED ORGANS OF NEW ZELAND MALE RABBITS IN NORTH SINAI

Twenty five New Zealand White (NZW) rabbit bucks, (aged 9 months and weighed 3.28± 0.1kg) were divided randomly into five comparable groups (5 each).Rabbits in the 1st group were fed pelleted control dietwithout medicinal plants (CO), while those in the 2nd, 3rd, 4th and 5th groups were fed ­the pelleted diet containing 3% rosemary (RS), 3% marjoram (MR), 1.5% ginger root (ZR) and 2.5% mixture (MX) of these medicinal plants, respectively. Results showed that rabbit bucks of the ZR group had lower (P<0.05) blood glucose level compared with the control, but did not differ significantly from the other treatments. Serum total protein increased (P<0.05) in RS, MR and MX when compared with control group, but ZR did not differ significantly from the control. Mean concentrations of serum albumin (AL) and AL/GL ratio did not differ significantly due to treatments. Rabbit fed diets containing rosemary, marjoram, ginger or their mixture showed lower (P<0.05) concentrations of plasma triglycerides than those fed the control diet. Rabbit fed ZR and MX had the lowest (P<0.05) level of cholesterol concentration, compared with for CO, RS and MR groups. Serum testosterone levels were significantly (P≤0.05) higher in rabbit bucks of ZR and MX groups compared with CO, RS and MR, respectively.Semen Ejaculate volume was significantly higher in MR, ZR and MX groups compared with the control but RS group did not differ significantly from the control. Means of mass motility were higher (P≤0.05) in treatment groups of RS, ZR and MX than the control but MR group did not differ from the control. The effect of dietary treatment on sperm cell concentration was significant. The ZR group had the highest mean followed by MX, RS, CO and MR groups. The histological examination of testicular sections showed that the elongated spermatids were more apparent in the ZR group and Leyding cells were well-developed in the MX group. The histological examination of liver sections showed marked increase in the size and number of bile ducts and many bi-nucleated hepatocytes and sinusoidal dilatation appeared in RS group. MR group showed an increase in the number of bile ducts, mononuclear phagocytic cells and sinusoid dilatation was observed. ZR group showed a lymphocytic infiltration was surrounding the bile duct and increase in the number of Kupffer cells. In MX group no magnificent changes were observed except that dilatation of the bile duct also no change in the sinusoidal size was observed. The histological examination of kidney sections of different experimental groups showed a congestion of some blood vessels in the MR, ZR and MX groups. Conclusively, the medicinal plants, Rosemary (Rosmarinus officinalis), Marjoram (Origanum majorana), Ginger root (Zingiber officinale) and their mixture could be added to NZW rabbit diets within the limits recorded in this study to improve semen characteristics and reproductive performance of NZW rabbit bucks without any detrimental effects on their health.

Optimal line of hepatotomy for left lateral living donor liver transplantation according to the anatomical variations of left hepatic duct system.

Multiple duct anastomoses during LLS transplantation increase the incidence of biliary complications. The optimal plane of hepatotomy that results in the least number of bile ducts at the surface was investigated according to LHD variations. Ducts of 30 human livers were injected with resin and LHD branching on 3D-CT reconstructions were analyzed. Ducts on the virtual hepatotomy surface were estimated in three splitting lines. Variations with subtypes were described. Ia (66.7%): ducts from segments (S.) II-III form a common trunk and S.IV duct joins it. Ib (10%): common trunk formed by ducts from S.II-S.III while S.IV duct joins the common hepatic duct. IIa (16.67%): S.IV duct drains into S.III duct. IIc (3.33%): S.IV duct drains into both S.II and S.III ducts. III (3.33%): trifurcation of S.II, S.III and S.IV ducts. When the virtual hepatotomy line was on the FL, there was a single duct for the anastomosis in 30% of cases but two, three, or four ducts in 53.3%, 10%, and 3.3%, respectively. Division 1 cm to the right of the FL resulted in one duct (70%), but S.IV duct injury may occur. LLS hepatotomy should not necessarily be performed along the FL. Variations must be taken into consideration to minimize the number of biliary anastomoses during liver implantation.

Biliary complications after pediatric liver transplantation: Risk factors, diagnosis and management.

The expanded indications of partial grafts in pediatric liver transplantation have reduced waiting list mortality. However, a higher morbidity is observed, including an increased rate of biliary complications (BCs). Factors such as the type of graft, the preservation methods applied, the donor characteristics, the type of biliary reconstruction, and the number of bile ducts in the liver graft influences the occurrence of these complications. Bile leaks and strictures comprise the majority of post-transplant BCs. Biliary strictures require a high grade of suspicion, and because most children have a bileo-enteric anastomosis, its diagnosis and management rely on percutaneous hepatic cholangiography and percutaneous biliary interventions (PBI). The success rates with PBI range from 70% to 90%. Surgery is reserved for patients who have failed PBI. BCs in children after liver transplantation have a prolonged treatment and are associated with a longer length of stay and higher hospital costs. However, with early diagnosis and aggressive treatment, patient and graft survival are not significantly compromised.

Impact of Early Biliary Complications On Long Term Graft and Patient Survival After Right Lobe Living Donor Liver Transplantation (LD).

Background: There is a significantly higher incidence of biliary complications after LD compared to deceased donor liver transplantation. It is not clear if this negatively impacts long term patient and graft survival after LD. Methods: Analysis of a prospectively maintained database of 120 LD with >2 year follow up at a single center. Actuarial patient and graft survival rates were estimated using Kaplan Meier methods. Results: 26 patients had biliary complications (bile leak=24;Stricture=12) and all occurred within 90 days post tx. 20 patients were managed by endoscopic stenting (58 procedures in 20 pts);6 required surgical exploration(none in the last 5 yrs). Time-to-complete resolution of leak and stricture was 37 and 82 days respectively. Number of bile ducts, duct to duct vs.roux-en-y, interrupted vs. continuous anastomosis, graft weight were not statistically associated with biliary complications. By Multivariate Cox Proportional Hazards regression model cardiovascular disease pretransplant (p=0.042) and HCV etiology (p=0.026) were associated with higher incidence of biliary complications. The one-year graft survival was lower (88% vs.97%; p=0.034) in biliary complications group, however there was no statistical difference in the 5-year graft survival (p=0.380).Figure: No Caption available.The patient survival was similar between the groups(p=0.331).Figure: No Caption available.Conclusions: Early Biliary complications result in lower 1-year graft survival but do not negatively impact long term patient survival after living donor liver transplantation.

α1- and α5-containing Laminins Regulate the Development of Bile Ducts via β1 Integrin Signals

Signals derived from basal lamina components are important for developing three-dimensional architecture of epithelial tissues. Laminins consisting of α, β, and γ subunits in basal lamina play pivotal roles in the formation and maintenance of epithelial tissue structures. However, it remains unclear which laminin isoforms transmit signals and how epithelial cells receive them to regulate multiple developmental processes. In three-dimensional culture of a liver progenitor cell line, Hepatic Progenitor Cells Proliferating on Laminin (HPPL), the cells establish apicobasal polarity and form cysts with a central lumen. Neutralizing antibody against β1 integrin blocked the formation and maintenance of the cyst structure, indicating that β1 integrin signaling was necessary throughout the morphogenesis. Although the addition of α1-containing laminin, a ligand of β1 integrin, induced cyst formation, it was dispensable for the maintenance of the cyst, suggesting that HPPL produces another ligand for β1 integrin to maintain the structure. Indeed, we found that HPPL produced α5-containing laminin, and siRNA against laminin α5 partially inhibited the lumen formation. In fetal liver, p75NTR(+) periportal fibroblasts and bile duct epithelial cells, known as cholangiocytes, expressed α1- and α5-containing laminins, respectively. In laminin α5 KO liver, cholangiocytes normally emerged, but the number of bile ducts was decreased. These results suggest that α1-containing laminin is sufficient as a component of the basal lamina for the commitment of bipotential liver progenitors to cholangiocytes and the apicobasal polarization, whereas α5-containing laminin is necessary for the formation of mature duct structures. Thus, α1- and α5-containing laminins differentially regulate the sequential events to form epithelial tissues via β1 integrin signals.

RhGH Attenuates Ischemia Injury of Intrahepatic Bile Ducts Relating to Liver Transplantation

To study the effect of rhGH administration on intrahepatic cholangiocytes relating to liver transplantation with ischemia of hepatic artery, and ultimately, clarify pathologic mechanism of the injury. Rat orthotopic autologous liver transplantation was performed first. Three hours later, the rats were grouped as followed: HAL (hepatic artery ligation) group; HAL + rhGH (hepatic artery ligation followed by rhGH administration) group; CON (without hepatic artery ligation) group. Specimen was collected after 7 d. ALT and ALP of serum were measured. The pathologic changes of bile ducts of liver tissue were observed. The number of bile ducts and blood vessels in portal area were counted. Immunochemistry for VEGF, VEGFR-2, VEGFR-3, GHR, and IGF-1R of intrahepatic cholangiocytes was performed. Cholangiocytes apoptosis was evaluated by TUNEL analysis. Cholangiocytes proliferation was evaluated by PCNA immunolabeling. ALT and ALP of HAL + rhGH group were significantly ameliorated compared with untreated animals (P < 0.05). ALT and ALP of HAL group were significantly higher compared with CON group (P < 0.05). In HAL group, the main injury of bile ducts was not reversible, whereas it was reversible in CON and rhGH groups. In HAL group, the number of bile ducts in portal area decreased, while the number of bile ducts not accompanying blood vessels increased (P < 0.05). In rhGH group, the number of bile ducts in portal area increased, while the number of bile ducts accompanying blood vessels increased compared with HAL group (P < 0.05). The expression of VEGF, VEGFR-2, VEGFR-3, GHR, and IGF-1R was significantly lower in HAL group than in CON group (P < 0.05). Following administration of rhGH to HAL rats, the expression of VEGF, VEGFR-2, VEGFR-3, IGF-1R, and GHR was significantly higher (P < 0.05). Administration of rhGH prevented increase in cholangiocytes apoptosis induced by HAL (P < 0.05). Administration of rhGH promoted increase in cholangiocytes proliferation held by HAL (P < 0.05). Administration of rhGH appears to attenuate ischemia injury of intrahepatic bile ducts relating to liver transplantation. This function is partly related to the capacity that rhGH inhibits the apoptosis of intrahepatic cholangiocytes and prompts the proliferation and angiogenesis by increasing the expression of VEGF, VEGFR2, VEGFR3, GHR, and IGF1-R.

Histologic features of the portal plate in extrahepatic biliary atresia and their impact on prognosis—a Danish study

Background/Purpose The aims of this study are as follows: 1. To evaluate the outcome of the patients with extrahepatic biliary atresia (EHBA), operated at the Rigshospitalet, Denmark, during the last 24 years. 2. To relate the histologic findings of the portal plate to the outcome. 3. To compare the histologic findings in the portal plate of children affected by EHBA with those found at the porta hepatis in age-matched children dead from other causes. Method From 1979 to 2003, 57 children have been operated by the Kasai procedure. Only 40 of these have had their portal plate removed for histologic examination. We divided the patients according to clinical outcome into a successful and a failure group and compared the histologic features of the portal plates in the 2 groups. Afterward, the portal plate histology from EHBA was compared with the porta hepatis area from patients dead from other causes. Results A significant difference between the success and the failure group was found with regard to the number of bile ducts, the maximal length measurable in any direction for bile duct structures, and the proliferation, but not for any type of diameter. The normal portal plate was different from the portal plate of children with biliary atresia by always having 2 large biliary structures and the cells being mucinous and columnar in the largest bile ducts. Only 1 of 4 normal portal plates showed signs of proliferation. There was no significant difference between the normal and the success group with respect to the number, maximal length, and proliferation of the bile ducts. A difference in the diameter between the normal group and the entire EHBA group was significant for the maximal internal diameter but not for the other types of diameter measurements. Conclusion The present study shows the following: 1. In portal plates from patients with EHBA, the longest measurable length of the bile duct structures, the number of bile duct structures, and less proliferation are positively correlated to prognosis. 2. Patients with EHBA have a more favorable prognosis when the histology of the portal plate resembles the normal portal plate regarding the number of bile ducts, proliferation, and maximal measurable length.

Bile duct to portal space ratio and ductal plate remnants in liver disease of infants aged less than 1 year

To validate the bile duct to portal space ratio as an independent factor useful for the prognosis of neonatal liver disease. We assessed the maturation of the intrahepatic bile duct system (IBDS) in 87 consecutive infants aged less than 1 year undergoing non-subcapsular, adequate (at least six portal tracts), liver needle biopsies because of hepatomegaly and/or cholestasis. The maturation of the IBDS was evaluated by immunohistochemistry with an antibody directed to cytokeratin 7 (CK7), a biliary-type intermediate filament of the cytoskeleton, and a schema showing the IBDS remodelling. We used five categories to fit the different patterns of the IBDS remodelling using the ratio between the number of bile ducts and the number of portal tracts (BD/PT) and the presence of abnormal reaction patterns (marked intra-acinar pseudorosettes and/or periportal ductular proliferation): (A) abnormal reaction patterns with any BD/PT; (B) BD/PT = 0; (C) 0.1 < or = BD/PT < 0.5; (D) 0.5 < or = BD/PT < 0.9; and (E) BD/PT > 0.9 (B-E categories: no abnormal reaction patterns). Further, we evaluated cholestasis, portal fibrosis (PF), portal inflammation (PI), giant cell transformation (GCT), and extramedullary haematopoiesis (EMH). We identified A-E categories in 24, 14, 17, 8, and 24 biopsies, respectively. B and C categories were frequently observed in biliary atresia (BA), A category in neonatal hepatitis (NH), A-C categories in paucity of intrahepatic bile ducts (PIBD), and E category in 'other liver diseases' (OLD). Cholestasis, PI, GCT, and EMH were more frequent in A and C, while PF was variably seen in all categories. The lowest survival rate occurred in B (Kaplan-Meier estimator). (1) Biliary epithelial cell patterns recapitulate the primitive stages of the IBDS maturation; (2) abnormal reaction patterns occur mainly in NH, whilst BD/PT < 0.5 in BA; and (3) lack of intrahepatic bile ducts in infants aged less than 1 year is an adverse prognostic factor independent from aetiology of neonatal liver disease.

Peroxisome proliferators-activated alpha agonist treatment ameliorates hepatic damage in rats with obstructive jaundice: an experimental study

BackgroundPeroxisome proliferators-activated receptor alpha (PPARα) activation modulates cholesterol metabolism and suppresses bile acid synthesis. This study aims to evaluate the effect of short-term administration of fenofibrate, a PPARα agonist, on proinflammatory cytokines, apoptosis, and hepatocellular damage in cholestasis.MethodsForty male Wistar rats were randomly divided into four groups: I = sham operated, II = bile duct ligation (BDL), III = BDL + vehicle (gum Arabic), IV = BDL + fenofibrate (100 mg/kg/day). All rats were sacrificed on 7th day after obtaining blood samples and liver tissue. Total bilirubin, aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP), gamma-glutamyl transferase, (GGT), tumor necrosis factor alpha (TNF-α), interleukin 1 beta (IL-1 β), and total bile acid (TBA) in serum, and liver damage scores; portal inflammation, necrosis, bile duct number, in liver tissue were evaluated. Apoptosis in liver was also assessed by immunohistochemical staining.ResultsFenofibrate administration significantly reduced serum total bilirubin, AST, ALT, ALP, and GGT, TNF-α, IL-1 β levels, and TBA (P < 0.01). Hepatic portal inflammation, hepatic necrosis, number of the bile ducts and apoptosis in rats with BDL were more prominent than the sham-operated animals (P < 0.01). PPARα induction improved all histopathologic parameters (P < 0.01), except for the number of the bile duct, which was markedly increased by fenofibrate therapy (P < 0.01).ConclusionShort-term administration of fenofibrate to the BDL rats exerts beneficial effects on hepatocellular damage and apoptosis.

MARKED BILE DUCT PROLIFERATION IN MICE HETEROZYGOUS FOR LUNATIC FRINGE AND JAG1 MUTATIONS

Background: The Notch signaling pathway is involved in cell fate determination in many organs. JAG1, encoding a ligand in the Notch pathway, has been identified as the disease gene for Alagille Syndrome (AGS), characterized by bile duct paucity and other manifestations. Mice heterozygous for mutations in both the Notch2 receptor and Jag1 ligand have a phenotype similar to AGS. Notch signaling is regulated by three mammalian Fringe genes. These genes encode glycosyltransferases that modify the activation of the Notch receptor by the ligand. To examine the role of the Lunatic Fringe (Lfng) gene in modifying Jag1 ligand function, we carried out genetic analysis of double heterozygous mice (Jag1tm1Grid/+; Lfngtm1Rjo/+, abbreviated as J1LF). Methods: Liver sections from both J1LF and control mice were examined by H&E staining. Bile ducts were identified by cytokeratin staining, and blood vessels were stained with Factor VIII antibody. Mice were evaluated as newborns and 5-week old adults. Quantitative real time RT-PCR for Hes1, a downstream effector in the Notch pathway, was performed to measure Notch signaling. Results: Adult J1LF mice had a significant increase in the number of bile ducts in the portal tracts, when compared to their single heterozygous and wild type littermates. Numbers of blood vessels in portal tracts were not significantly different between J1LF and control mice. Gross examination revealed that the extrahepatic biliary tree was intact. In adult J1LF livers with bile duct proliferation, real time PCR demonstrated a 4 to 8 fold increase in Hes1 expression levels over wild type and single heterozygous controls. However, in the newborn period, J1LF mice did not display a significant difference in the numbers of bile ducts. Conclusions: Jag1/Lfng double heterozygous mice have a striking proliferation of bile ducts in the adult liver. These differences arise postnatally, pointing to a role for Lfng in regulating Jag1-Notch signaling in bile duct growth and maturation and LFNG as a potential modifier gene for AGS.

Biliary epithelial cell response to cholestasis

T ENTIRE biliary tract is lined by a single layer of epithelial cells that, besides purely conveying functions, modify canalicular bile composition through vectorial transport of water, electrolytes and macromolecules. This transport activity depends upon the polarized distribution of plasma membrane proteins. Fluid ductular secretion represents 20 to 40% of total bile flow in humans, and is stimulated essentially by secretin and the vasoactive intestinal polypeptide. Secretin via a CAMP transient increase, stimulates the combined activities of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel and of the anion exchanger (AE-2), both of which are located in the apical domain of biliary epithelial cells. Their activation results in water and bicarbonate secretion, leading to bile dilution and alkalinization. Biliary epithelial cells are also committed to absorption. The apical sodium-dependent bile acid transporter (ASBT) is present in the apical domain of biliary epithelial cells, where it may provide a cholehepatic shunt pathway for conjugated bile acids via the peribiliary vascular plexus (1). A general feature of cholestatic liver injury is the increase in bile ductular structures, referred to as “ductular reaction”. Depending on the type of injury, the ductular reaction may be “typical” and result from proliferation of pre-existing bile duct epithelial cells, or “atypical” and resplt from metaplasia of hepatocytes or from hepatic stem cell proliferation (2). The typical ductular reaction induced by acute biliary obstruction is associated with a marked increase in ductular secretion, and may therefore be regarded as an escape route for bile flow. Secretin-induced ductal secretion is exacerbated due to both an increase in the number of bile ducts and an up-regulation of secretin receptors (3). The cholehepatic shunt pathway may also be