Bile Duct Hyperplasia Research Articles

Hepatobiliary Events in Migraine Therapy with Herbs-The Case of Petadolex, A Petasites Hybridus Extract.

Petadolex®, a defined butterbur extract has clinically proven efficacy against migraine attacks. However, spontaneous reports indicate cases of herbal induced liver injury (HILI). While most HILI patients presented mild serum biochemistry changes (<3 ULN, dose range 50 to 225 mg/day; treatment duration 4–730 days) nine developed severe HILI (average time-to-onset 103 days, ALT-range 3–153; AST 2–104-fold ULN). HILI cases resolved after medication withdrawal though two patients required liver transplantation. Liver biopsies revealed an inconsistent injury pattern, i.e. necrosis, macrovesicular steatosis, inflammation, cholestasis, and bile duct proliferation. Causality assessment rated 3 cases likely, 13 possible, 8 unlikely and 24 as unclassifiable/unclassified. Note, 22 patients reported hepatotoxic co-medications especially during periods of pain. A no-observable-adverse-effect-level at 15-fold of the maximal clinical dose (3 mg/kg/day MCD) was established for rats. At >45 and 90-fold MCD bile duct hyperplasia was observed but could not be confirmed in an explorative minipig study at 218-fold MCD. Human hepatocyte studies at 49-fold Cmax serum petasins (=active ingredient) and therapeutic Ibuprofen, Paracetamol and Naratriptan concentrations evidenced liver transaminase and CYP-monooxygenase changes. Collectively, Petadolex® HILI cases are rare, idiosyncratic and frequently confounded by co-medications. A physician-supervised self-medication plan with herbs and pain relief medication is needed to minimize risk for HILI.

Molecular characterization and histopathological studies on Fasciola gigantica in Mithun (Bos frontalis)

Mitochondrial DNA sequence of the sub-unit 1 of cytochrome c oxidase (Cox1) and ribosomal DNA sequence of the first internal transcribed spacer (ITS-1) of Fasciola collected from mithun and cattle from Arunachal Pradesh, India were characterized at molecular level in order to identify the species of parasite. Based on sequence and phylogenetic analysis, the identity of the parasite was confirmed as Fasciola gigantica in mithun as well as from cattle. In order to know histopathological alteration in Fasciola infection, a histopathological study was performed on eight liver specimens, out of 110 animals studied during the period from 2010-2016. Histopathological examination of F.gigantica infected liver showed extensive fibrous connective tissue proliferation with necrosis of hepatocytes and infiltration of polymorphonuclear cell. There was evidence of migratory tracts of parasites with losing of normal lobular hepatic architecture. Bile duct proliferation was followed by congestion of portal vein with perivascular cuffing with surrounding degeneration of hepatocytes. The hepatocytes showed pyknosis with hyperplasia of bile duct.

Post‐Natal Growth and Liver Immune Homeostasis are Post‐Transcriptionally Regulated by Tristetraprolin Family of RNA Binding Proteins

Tristetraprolin family of RNA binding proteins, including tristetraprolin (TTP), zinc finger protein 36 like 1 (ZFP36L1), and zinc finger protein 36 like 2 (ZFP36L2) regulate mRNA levels by binding to AU‐rich elements on the 3′ untranslated regions of specific mRNAs and enhancing their turnover. While the three proteins behave similarly under biochemical conditions, they appear to have diverse physiological functions. This is apparent from the global deletion of the three proteins in mice. While the global loss of TTP results in systemic inflammatory syndrome, global loss of ZFP36L1 is embryonic lethal, and the global loss of ZFP36L2 results in early post‐natal mortality. Here, we sought to understand the physiological functions of the three proteins in liver by conditional liver specific deletion using Cre‐loxP technology. While the independent deletion of the three proteins did not result in any aberrant phenotype, simultaneous deletion resulted in post‐natal growth defect, chronic active portal hepatitis, and bile duct hyperplasia. Transcriptomic analysis showed small numbers of gene expression changes in the liver of ZFP36L1 knockout, no changes in the livers of TTP and ZFP36L2 knockout, and many changes in the liver of mice deleted of the three proteins simultaneously. The changes included perturbation of genes that regulate post‐natal growth and several inflammatory mediators, some of which are novel potential liver‐specific targets of TTP family proteins. Interestingly, one allele of either ZFP36L1 or ZFP36L2 was sufficient to protect the mice from developing this phenotype. However, one allele of TTP was only partially protective. In summary, our results suggest that post‐transcriptional regulation of gene expression plays an important role in post‐natal growth and liver immune homeostasis and that TTP family proteins regulate these physiological functions in both specific and redundant manner.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Histological Analysis of Lysophosphatidic Acid Receptor 3 Deficient Zebrafish

Lysophosphatidic acid (LPA) is a small bioactive phospholipid enriched in serum and exerts diverse cellular functions through activation of multiple G Protein‐coupled receptors, LPA1‐LPA6. Our previous studies demonstrated that lysophosphatidic acid receptor 3 (LPA3) is critical for megakaryopoiesis and erythropoiesis in zebrafish. However, the roles of LPA3 during the development of other organs of zebrafish remain unclear. Here, we established a zebrafish strain with targeted disruption of lysophosphatidic acid receptor 3 (Lpar3) gene by Transcription Activator‐like Effector Nuclease (TALEN). Paraffin‐section and hematoxylin‐eosin staining were performed for histological analysis to clarify the effects of LPA3 in different organs of adult zebrafish. Histological analysis results showed a shrinking of hematopoietic tissue in kidney marrow of LPA3 deficient fish, implies essential roles of LPA3 in hematopoiesis. In addition, bile duct hyperplasia and mature spermatids loss were also observed in LPA3 deficient fish. Nevertheless, no obvious morphological differences in eyes, thymus, heart, and spleen were observed. Our observation provides tissues profiles of LPA3−/− animal and possible application of LPA3 deficient zebrafish as a disease model in the future.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

A novel prognostic model for diagnosing atypical bile duct hyperplasia in patients with intrahepatic lithiasis.

There is no specific method for the preoperative diagnosis of atypical bile duct hyperplasia, which is a precursor of cholangiocarcinoma. This study aimed to create a new model for diagnosing atypical bile duct hyperplasia based on routine laboratory tests in patients with intrahepatic lithiasis.The new diagnostic model was developed with a derivation cohort that included 375 patients with intrahepatic lithiasis. Clinical and pathological data were retrospectively collected. Prognostic factors were evaluated with univariate and logistic regression analyses. The validation cohort included 136 patients who were retrospectively screened to quantify the model's predictive value.Age and Carbohydrate Antigen 19-9 (CA-199) were revealed to be diagnostic indicators of atypical bile duct hyperplasia in patients with intrahepatic lithiasis. The new diagnostic model was created with the formula: -6.612 + (0.002 × CA-199) + (0.072 × Age). The area under the receiver operating curve of the model was 0.721. With 0.25 as the cutoff point, the sensitivity and specificity of this model in the derivation cohort were 13.9% and 95.9%, respectively. In the validation cohort, these values were 28.5% and 88.7%, respectively. The novel model has an acceptable and stable ability to predict atypical hyperplasia in the intrahepatic bile duct.This novel model provides a simple system for diagnosing atypical bile duct hyperplasia before surgery in patients with intrahepatic lithiasis.

CD24 regulates Ly6Chi macrophages and BDL-induced liver fibrosis

Objective To investigate the regulatory effects of CD24 on Ly6Chi macrophages in liver and its influences on bile duct ligation (BDL)-induced hepatic fibrosis in mice. Methods Liver fibrosis was induced in wild-type (WT) and CD24-/- mice by surgical ligation of the biliary duct. Levels of alanine amino transferase (ALT) in serum were detected and liver sections were stained with haematoxylin and eosin (H&E) staining to assess the severity of liver injury. Sirius Red staining was used to observe the degree of liver fibrosis. Real-time PCR was performed for detecting the expression of hepatic fibrosis-related markers and TGF-β1 at mRNA level. The percentage of macrophages and the number of TGF-β1-producing macrophages were measured by flow cytometry. Results BDL-induced liver fibrosis was exacerbated in CD24-/- mice than in WT mice as demonstrated by more serious hyperplasia in bile duct, more inflammatory infiltration at the portal area and higher levels of ALT in serum. Results of Sirius red staining also showed that the liver fibrosis was more severe in CD24-/- mice than in WT mice. Moreover, α-SMA and collagen typeⅠ alpha 1 (Col1a1) were significantly upregulated in CD24-/- mice. Flow cytometry analysis revealed that CD24 was highly expressed by hepatic macrophages in BDL-induced WT mice, and the percentages of hepatic macrophages were significantly elevated in CD24-/- mice compared with those in WT mice. Further analysis revealed that the percentages of Ly6Chi hepatic macrophages in CD24-/- mice were higher than those in WT mice, but there was no significant difference in the percentages of Ly6Clo macrophages. The expression of TGF-β1 at mRNA level was increased in CD24-/- mice as compared with that in WT mice after BDL. Moreover, intracellular staining showed that Ly6Chi hepatic macrophages in CD24-/- mice secreted more TGF-β1 than the macrophages in WT mice. Conclusions CD24 might attenuate the BDL-induced liver fibrosis in mice via regulating the percentage of hepatic Ly6Chi macrophages and the secretion of TGF-β1. Key words: CD24; Bile duct ligation (BDL); Liver fibrosis; Ly6Chi hepatic macrophage; TGF-β1

Hepatic Encephalopathy Secondary to Chronic Liver Lesions Caused by Crotalaria incana in a Bovine

Background: Crotalaria spp. Poisoning induces liver or pulmonary disease. C. mucronata, C. juncea, C. spectabilis, and C. retusa are the Crotalaria spp. inducing spontaneous intoxication in livestock in Brazil. C. mucronata and C. juncea are associated with interstitial pneumonia, while C. retusa and C. specatabilis induce hepatotoxicosis. The toxic principle in Crotalaria spp. are dehydropyrrolizidine alkaloids and their N-oxides. C. incana poisoning to livestock is rarely documented. This paper reports the clinical signs and pathological findings of a case of Crotalaria incana poisoning in a steer. The chemical finding of a potential toxic dehydropyrrolizidine alkaloid in the plant is documented for the first time.Case: The affected bovine was part of a herd of 80 two-year-old steers that were transferred from Property 1 to Property 2, 30 days prior to the event. In the pasture of Property I - where the steers were held for 6 months - there was a heavy infestation by a Crotalaria species with signs of being consumed by the steers. The plant was identified as Crotalaria incana at the Botanical Laboratory of the Federal University of Mato Grosso do Sul and a voucher specimen was filed there under the register GCMS 51169. Two days after entering Property 2 one of the steers became depressed, staggering, and in poor body condition. With time, the steer became oblivious to the environment and died 20 days after the onset of the clinical signs. Significant necropsy findings were limited to the liver which was markedly enlarged and with rounded edges. The hepatic cut surface was mottled with dark red and extensive orange areas of discoloration. The gallbladder was distended and the bile was inspissated. Microscopically, in the liver, there was fibrosis, bile duct hyperplasia and hepatocellular megalocytosis. The Glisson’s capsule was markedly thickened by fibrosis. In the brain, there was vacuolation of myelin sheaths (status spongiosus), moderate gliosis, and rare Alzheimer type 2 astrocytes.Discussion: The diagnosis of intoxication by C. incana was based on clinical signs and pathological changes and the evidence of the plant being consumed. The high infestation of C. incana in the pasture, the high proportion of dead matter and the low concentration of viable pasture favored the ingestion of C. incana. Spongy degeneration, a typical lesion of hepatic encephalopathy, was observed in the several areas of the brain. Cirrhosis of the liver as seen in the case of this report, results in elevated ammonia levels in the blood - and eventually in the brain – and ammonia toxicity causes hepatic encephalopathy. Morphological changes in hepatic encephalopathy in the central nervous system of human beings and horses centers on astrocytes which undergo Alzheimer type 2 change developing an enlarged, pale nuclei with a rim of chromatin and prominent nucleoli. Astrocytes pairs and triplets are seen, and, in severe cases, astrocytic nuclei may become lobulated and contain glycogen. Chemical analysis of C. incana was accomplished by acid-base extraction with zincreduction of N-oxides and quantitation by spectrometry. Quantitative analysis was achieved by liquid chromatography-mass spectrometry. To confirm the identity of the alkaloid in the sample of C. incana the isolated alkaloid was compared to a standard sample of usaramine previously isolated, and its identity verified by magnetic resonance spectroscopy analysis. The 1,2-dehydropyrrolizidine alkaloid usaramine, as well as its N-oxide, were identified as the major alkaloids in in C. incana. The concentration found in the seed was consistent with that previously reported. This is a new finding on the toxic principle of C. incana. Keywords: cattle diseases, poisonous plants, pathology, hepatotoxicity, Crotalaria incana, chronic liver failure.

Haemato-biochemical and pathomorphological observations in paramphistomosis of domestic ruminants in Chitt oor district, Andhra Pradesh

A total of 671 samples (blood, serum and tissues) were collected from 382 domestic ruminants. Blood samples were subjected for PCV, Hb and RBC counts. No signifi cant difference was noted between paramphistomosis infected (PI) and control group (CG) of cattle and sheep with regard to PCV, Hb and RBC values, but in goat signifi cant difference was observed. In DLC, statistically, no signifi cant difference was observed in immature amphistomosis (IA), PI and CG of cattle. In IA and CG of sheep, neutrophil and monocyte values showed signifi cant alterations, but in goats it was in neutrophil and lymphocytes. Biochemical tests revelaed low levels of total serum proteins and albumin in IA infected cattle, sheep and goats. Gross and histopathological changes in rumen, reticulum and bile duct of animals of PI group were observed. The adult amphistomes invading the rumen mucosa by sucker and plugging of mucosa of bile duct were noticed in cattle. In the bile duct of cattle, massive and extensive infiltration of plasma cells, lymphocytes and monocytes in between acinar structures was recorded. Hyperplasia of bile duct, degenerative changes and necrotic changes in acinar mucosal lining epithelial cells were appreciated prominently. In IA, duodenum of sheep revealed thickened and edematous; mucosal surface with severe congestion, petechial haemorrhages and necrosis. Histopathological studies of IA infected duodenum of sheep showed plugging of mucosa with sucker and extensive infi ltration of macrophages and plasma cells deep into the mucosa.

Seasonal development of the coccidian parasite Goussia bayae and hepatobiliary histopathology in white perch Morone americana from Chesapeake Bay.

The coccidium Goussia bayae infects the gallbladder and bile ducts of white perch Morone americana from Chesapeake Bay, USA. Seasonal changes in coccidian infections were analyzed from bile specimens of 1588 fish from the Choptank River during 2016-2018 using wet mount preparations with a Sedgwick-Rafter counting chamber. Histopathology of the gallbladder and liver was analyzed from a subset (n = 480) of these fish. Maximum parasite prevalence (100%) and intensities in the gallbladder occurred during the fish spawning season in March and April. Asynchronous coccidian development and prevalence of infections in fish increased gradually during autumn and winter, but coccidian intensity increased sharply 2-4 wk prior to the onset of fish spawning activity and decreased after spawning activity concluded. Sporulation was internal, and the gallbladder was the primary reservoir for oocysts. Two previously undescribed species of coccidia were observed in the intestine. Lesions in the gallbladder were rare and included cholecystitis and epithelial necrosis. Intrahepatic bile duct lesions were more common and included distension, cholangitis, epithelial erosion and necrosis, cholestasis, hyperplasia, and neoplasia. Cholangitis and necrosis of intrahepatic bile ducts were significantly associated with coccidial infections, while plasmodia of a myxosporean (spore morphology consistent with the genera Myxidium and Zschokella) were significantly associated with bile duct hyperplasia. Biliary neoplasia included cholangiomas (5% prevalence) and cholangiocarcinomas (1% prevalence). No association was detected between G. bayae and biliary neoplasms, but an association may exist between these lesions and the myxosporean plasmodia.

The effects of astaxanthin on liver histopathology and expression of superoxide dismutase in rat aflatoxicosis.

The metabolism of aflatoxin B1 (AFB1) generates reactive oxygen species (ROS) that destroys hepatocytes. Meanwhile, astaxanthin (AX) is known to have stronger antioxidative activity than other carotenoids. This study aimed to investigate hepatoprotective role of AX from AFB1-induced toxicity in rat by histopathological study and immunohistochemistry of Cu/Zn-SOD (SOD1) which acts as the first enzyme in antioxidative reaction against cell injury from ROS. Twenty Wistar rats were randomly divided into 4 groups. The control and AFB1 groups were gavaged by water for 7 days followed by a single DMSO and 1 mg/kg AFB1, respectively. The AXL+ AFB1 and AXH+ AFB1 groups were given of 5 mg/kg and 100 mg/kg AX for 7 days before 1 mg/kg AFB1 administration. The result showed significantly elevated liver weight per 100 g body weight in AFB1 group. The histopathological finding revealed vacuolar degeneration, necrosis, megalocytosis and binucleation of hepatocytes with bile duct hyperplasia in AFB1 group. The severities of pathological changes were sequentially reduced in AXL+AFB1 and AXH+AFB1 groups. Most rats in AXH+AFB1 group owned hypertrophic hepatocytes and atypical proliferation of cholangiocytes which are adaptive responses to severe hepatocyte damage. The SOD1 expression was also significantly higher in AXH+AFB1 group than solely treated AFB1 and AXL+AFB1 groups. In conclusion, AX alleviated AFB1-induced liver damage in rat by stimulating SOD1 expression and transdifferentiation of cholangiocytes in dose dependent manner.

A chronic toxicity study of diphenylarsinic acid in the drinking water of C57BL/6J mice for 52 weeks.

Diphenylarsinic acid (DPAA), a neurotoxic organic arsenical, is present in the groundwater and soil in some regions of Japan due to illegal dumping. The purpose of the present study was to evaluate the potential toxicity of DPAA when administered to mice in their drinking water for 52 weeks. DPAA was administered to mice at concentrations of 0, 6.25, 12.5, and 25 ppm in their drinking water for 52 weeks. There were no significant differences in final body weights between the control groups and the DPAA treatment groups in male or female mice. Relative liver weights were significantly increased in males treated with 25 ppm DPAA, and absolute liver weights were significantly decreased in female mice treated with 25 ppm DPAA. In female mice, cholangitis and simple bile duct hyperplasia were observed in the 12.5 and 25 ppm DPAA groups, and focal necrosis of hepatocytes was observed in the 25 ppm DPAA group. Proteomic analysis and Ingenuity Pathway Analysis identified 18 proteins related to hepatotoxicity that were overexpressed in the female 25 ppm group. The phase I metabolic enzyme CYP2E1 was one of these overexpressed proteins. Immunostaining confirmed high expression of CYP2E1 in the livers of females in the 25 ppm group. These results suggest that DPAA is toxic to the intrahepatic bile duct epithelium and hepatocytes in female mice and that CYP2E1 might be involved in DPAA-associated toxicity. The no-observed-adverse-effect levels of DPAA were 12.5 ppm (1.6 mg/kg bw/day) for males and 6.25 ppm (1.1 mg/kg bw/day) for females under the conditions of this study.

Ductular Reaction in Liver Diseases: Pathological Mechanisms and Translational Significances.

Ductular reaction (DR) is characterized by the proliferation of reactive bile ducts induced by liver injuries. DR is pathologically recognized as bile duct hyperplasia and is commonly observed in biliary disorders. It can also be identified in various liver disorders including nonalcoholic fatty liver disease. DR is associated with liver fibrosis and damage, and the extent of DR parallels to patient mortality. DR raises scientific interests because it is associated with transdifferentiation of liver cells and may play an important role in hepatic regeneration. The origin of active cells during DR can be cholangiocytes, hepatocytes, or hepatic progenitor cells, and associated signaling pathways could differ depending on the specific liver injury or animal models used in the study. Although further studies are needed to elucidate detailed mechanisms and the functional roles in liver diseases, DR can be a therapeutic target to inhibit liver fibrosis and to promote liver regeneration. This review summarizes previous studies of DR identified in patients and animal models as well as currently understood mechanisms of DR.

Hedgehog Signaling Modulates Interleukin-33-Dependent Extrahepatic Bile Duct Cell Proliferation in Mice.

Hedgehog (HH) signaling participates in hepatobiliary repair after injury and is activated in patients with cholangiopathies. Cholangiopathies are associated with bile duct (BD) hyperplasia, including expansion of peribiliary glands, the niche for biliary progenitor cells. The inflammation‐associated cytokine interleukin (IL)‐33 is also up‐regulated in cholangiopathies, including cholangiocarcinoma. We hypothesized that HH signaling synergizes with IL‐33 in acute inflammation‐induced BD hyperplasia. We measured extrahepatic BD (EHBD) thickness and cell proliferation with and without an IL‐33 challenge in wild‐type mice, mice overexpressing Sonic HH (pCMV‐Shh), and mice with loss of the HH pathway effector glioma‐associated oncogene 1 (Gli1lacZ/lacZ). LacZ reporter mice were used to map the expression of HH effector genes in mouse EHBDs. An EHBD organoid (BDO) system was developed to study biliary progenitor cells in vitro. EHBDs from the HH overexpressing pCMV‐Shh mice showed increased epithelial cell proliferation and hyperplasia when challenged with IL‐33. In Gli1lacZ/lacZ mice, we observed a decreased proliferative response to IL‐33 and decreased expression of Il6. The HH ligands Shh and Indian HH (Ihh) were expressed in epithelial cells, whereas the transcriptional effectors Gli1, Gli2, and Gli3 and the HH receptor Patched1 (Ptch1) were expressed in stromal cells, as assessed by in situ hybridization and lacZ reporter mice. Although BDO cells lacked canonical HH signaling, they expressed the IL‐33 receptor suppression of tumorigenicity 2. Accordingly, IL‐33 treatment directly induced BDO cell proliferation in a nuclear factor κB‐dependent manner. Conclusion: HH ligand overexpression enhances EHBD epithelial cell proliferation induced by IL‐33. This proproliferative synergism of HH and IL‐33 involves crosstalk between HH ligand‐producing epithelial cells and HH‐responding stromal cells.

Embracing the Dark Side: Computational Approaches to Unveil the Functionality of Genes Lacking Biological Annotation in Drug-Induced Liver Injury.

In toxicogenomics, functional annotation is an important step to gain additional insights into genes with aberrant expression that drive pathophysiological mechanisms. Nevertheless, there exists a gap on annotation of these genes which often hampers the interpretation of results and limits their applicability in translational medicine. In this study, we evaluated the coverage of functional annotations of differentially expressed genes (DEGs) induced by 10 selected compounds from the TG-GATEs database identified as high- or no-risk in causing drug-induced liver injury (most-DILI or no-DILI, respectively) using in vitro human data. Functional roles of DEGs not present in the most common biological annotation databases – termed “dark genes” – were unveiled via literature mining and via the identification of shared regulatory transcription factors or signaling pathways. Our results demonstrated that there were approximately 13% of dark genes induced by these compounds in vitro and we were able to obtain additional relevant information for up to 76% of those. Using interactome data from several sources, we have uncovered genes such as LRBA, and WDR26 as highly connected in the protein network that play roles in drug response. Genes such as MALAT1, H19, and MIR29C – whose links to hepatotoxicity have been confirmed – were identified as markers for the most-DILI group and appeared as top hits across all literature-based mining methods. Furthermore, we investigated the potential impact of dark genes on liver toxicity by identifying their rat orthologs in combination with their correlation to drug-induced liver pathologies observed in vivo following chemical exposure. We identified a set of important regulatory transcription factors of dark genes for all most-DILI compounds including E2F1 and JUND with supporting evidences in literature and we found Magee1 correlated with chemically induced bile duct hyperplasia and adverse responses at 29 days in rats in vivo. In conclusion, in this study we show the potential role of these poorly annotated genes in mechanisms underlying hepatotoxicity and offer a number of computational approaches that may help to minimize current gaps in gene annotation and highlight their values as potential biomarkers in toxicological studies.

Application value of perihilar surgery technique in the reoperation of biliary dilatation of central large intra- and extra-hepatic bile ducts above the hilar convergence

Objective To explore the application value of perihilar surgery technique in the reoperation of biliary dilatation of central large intra- and extra-hepatic bile ducts above the hilar convergence. Methods The retrospective cross-sectional study was conducted. The clinical data of 3 patients with biliary dilatation of central large intra- and extra-hepatic bile ducts above the hilar convergence who underwent the reoperation in the Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine from August 2017 to January 2018 were collected. All three patients had been collected a detailed previous surgical history. After preoperative lab and imaging examinations, evaluation of liver function, residual liver volume and general condition were done, then 3 patients underwent left hemihepatectomy + dilated right hepatic duct and extrahepatic bile duct resection, right anterior and posterior hepatic duct reconstructive surgery and Roux-en-Y anastomosis of the jejunum. The surgical procedures followed as: intra-abdominal adhesions separation, extrahepatic antergrade dissection of porta hepatis, transverse cutting the dilated extrahepatic bile duct, split the cantlie line, exposure of the hilar plate, left hemihepatectomy, dilated right hepatic bile duct resection, right anterior and posterior hepatic duct remodeling and biliary-enteric anastomosis. Observation indicators included: (1) surgical and postoperative recovery; (2) follow-up. Follow-up using outpatient examination and telephone interview was performed to detect general condition, complications, liver function and residual choledochal cysts up to May 2018. Results (1) Surgical and postoperative recovery: All the 3 patients underwent choledochal cysts resection + left hemihepatectomy + dilated right hepatic duct + right anterior and posterior hepatic duct reconstructive surgery and Roux-en-Y anastomosis of the jejunum using the perihilar surgery technique and extrahepatic anterograde combined by intrahepatic retrograde dissection method exposing portal hepatis. There was no perioperative death. The operation time and volume of intraoperative blood loss in 3 patients were 435 minutes, 490 minutes, 395 minutes and 250 mL, 300 mL, 200 mL, respectively. There was no intraoperative blood transfusion. Three patients had no bleeding and abdominal liver function. One patient with bile leakage and delayed gastric emptying at 1 week postoperatively received puncture drainage, gastrointestinal decompression, gastric lavage with hypertonic saline, acupuncture and total parenteral nutrition, then bile leakage was cured after 3-week therapy, gastric motility was improved after 5-week therapy, and then gastric tube was removed. The abdominal drainage tube was removed at 3 weeks postoperatively in 1 patient and at 1 week postoperatively in 2 patients. The postoperative gross specimen examinations showed intra- and extra-hepatic bile duct dilatation in 3 patients, including 2 combined with choledocholithiasis and 1 with left intrahepatic bile duct cancer. The postoperative pathological findings showed that 3 patients had intrahepatic bile duct cystic dilatation with chronic inflammation, peripheral small bile duct hyperplasia with inflammatory cell infiltration, 1 of which had intrahepatic intraductal papilloma with high grade intraepithelial neoplasia. One and 2 patients were discharged from hospital at 7 weeks postoperatively and 2 weeks postoperatively, respectively. (2) Follow-up: All 3 patients were followed up for 4-8 months. During the follow-up, patients had good general condition and no symptoms of cholecystitis such as abdominal pain, chills and fever, liver function was normal, and no residual bile duct cyst was found by enhanced scan of CT. Conclusion The perihilar surgery technique and extrahepatic anterograde combined by intrahepatic retrograde dissection method exposing portal hepatis for the treatment of biliary dilatation of central large intra- and extrahepatic bile ducts above the hilar convergence can increase the radical resection rate and surgical efficacy. Key words: Biliary dilatation; Perihilar technique; Central large hepatic bile duct above the hilar convergence; Extrahepatic anterograde combined by intrahepatic retrograde dissection method exposing portal hepatis; Surgical procedures, operative

Changes in Rats’ Liver Structure Induced by Zinc Oxide Nanoparticles and the Possible Protective Role of Vitamin E

Background: Oral ingestion of zinc oxide nanoparticles (ZnONPs) may lead to serious liver injury. Vitamin E (VE) is an important antioxidant factor that can reduce such damage. Aim: This study aimed to evaluate the possible changes that could take place in the liver of adult male albino rats after oral ingestion of ZnONPs and elucidate the potential protective role of VE against such damage. Material and Methods: Forty eight male albino rats were divided into four groups of 12 animals each. Group (1) served as control group and received normal saline. Group (2) “VE-treated” received 100 mg/kg/day of VE dissolved in normal saline by oral gavage for 21 days. Group (3) “ZnONPs-treated” received a daily dose of ZnONPs dispersed in the fresh sterilized physiological saline solution 1mg/kg for 5 constitutive days. Group (4) “concomitant ZnONPs and VE-treated” was pretreated with VE 100 mg/kg/day for 14 days followed by the same dose of ZnONPs as in group (3) for 5 days. The extent of hepatic damage was evaluated by histological and immunohistochemical examination of liver samples and serological analysis of liver enzymes. Results: Body weights and liver weights showed very highly significant decrease (P &lt;0.001) in the ZnONPs-treated group. The histological results in ZnONPs-treated group revealed congested dilated central veins and blood sinusoids, loss of normal arrangement of hepatocytes and most of hepatocytes showed marked vacuolated cytoplasm with darkly stained nuclei. Portal area affection was in the form of congested dilated portal veins with bile duct hyperplasia and cellular infiltration. There was an increase in the mount of blue stained collagen fibers around central veins together with strong positive reaction for Caspase 3 in ZnONPs-treated group. Similarly biochemical analysis indicated that the levels of serum aminotransferase (AST &amp;ALT) significantly increased in ZnONPs-treated group when compared with other groups. Rats pretreated with VE showed improvement of the histological findings and biochemical parameters. Conclusion: Ingestion of ZnONPs could be associated with serious liver affection and pretreatment with VE is suggested to induce some improvement of such deleterious changes.

Simultaneous lymphoid hyperplasia of gallbladder and bile duct mimicking carcinoma: a case report

Introduction: Lymphoid hyperplasia is a rare benign lymphoproliferative disorder. It can occur in various organs. However, lymphoid hyperplasia arising from extrahepatic bile duct and gallbladder simultaneously is extremely rare. Methods: A 72-year-old woman visited hospital with general weakness, dyspepsia and weight loss for 3 months. She had medical history of diabetes mellitus and depressive mood disorder and had been treated for liver abscess ten years ago. On physical examination, there was no icteric sclera and no tenderness in the upper abdomen. Viral hepatitis markers and all tumor markers were within normal limits. Magnetic resonance cholangiopancreatography (MRCP) showed 3cm length wall thickening and enhancement of suprapancreatic and intrapancreatic CBD, causing mild luminal narrowing and dilatation of upper biliary tract and also showed irregular wall thickening and enhancement of gallbladder body and fundus. Results: Under diagnosis of distal CBD cancer and gallbladder cancer, she underwent pylorus-preserving pancreaticoduodenectomy with routine lymph node dissection and s4b and S5 liver wedge resection. In operation finding, there were diffuse nodular sclerosing change from mid CBD to distal CBD and there were diffuse wall thickening of gallbladder body at liver bed side. Based on pathologic finding and immunohistochemical staining, lesion was diagnosed histologically as lymphoid hyperplasia. Conclusion: Lymphoid hyperaplasia of biliary system is a rare disease, and preoperative diagnosis is extremely difficult. Although it is benign condition, we should consider surgical excision for this lesion that cannot be excluded for malignancy.

Clinical significance and correlation of ductular reaction in hepatobiliary diseases

Ductular reaction is a kind of repair response to biliary and hepatocellular injury, and the pathological changes include lobular bile duct hyperplasia, matrix change and inflammatory cell infiltrations. In human liver, a ductular reaction originates from the activation and proliferation of hepatic progenitor cells. The activated hepatic progenitor cells, the intermediate hepatocytes and the reactive small bile duct cells together form the cell complex of ductular reaction, which has the dual characteristics of hepatobiliary system and co-expresses the hepatocyte and cholangiocyte antigen. Ductular reaction appears in all kinds of acute and chronic hepatobiliary injury, and plays an important role in liver regeneration and repair, fibrogenesis, cirrhosis formation, and hepatobiliary cancer occurrence and development. Understanding the pathological features and pathophysiological mechanisms of ductular reaction in various clinical hepatobiliary diseases and their role in liver regeneration can provide important suggestion for further exploring new-targeted treatment approach.

Poisoning of cattle by Senecio spp. in Brazil: a review

ABSTRACT: Poisoning of cattle by plants of the Senecio genus represents an important cause of death in cattle and has important economic repercussions in southern Brazil. This review is intended to provide a detailed review of Senecio spp. intoxication in cattle and addresses issues regarding the toxic principle and pathogenesis of the disease caused by these plants and the epidemiology, clinical signs, diagnosis, control and prophylaxis of the disease. Senecio brasiliensis is the main species associated with natural intoxication of livestock in Brazil, and the number of cases associated with the ingestion of S. madagascariensis is increasing. The toxic principle of Senecio spp. comprises the hepatotoxic alkaloids of the pyrrolizidine group (pyrrolizidine alkaloids, PAs). The resulting liver lesions are chronic and irreversible and result from the inhibition of hepatocellular mitosis. Deaths of adult cattle may occur both sporadically and in larger outbreaks over an extended period of time. In cattle raising, Senecio spp. are consumed mainly during the winter, when there is a dearth of forage; at this time the poisonous Senecio species are budding and contain high concentrations of PAs. Spontaneous Senecio spp. intoxication in cattle is a chronic condition that frequently involves acute clinical manifestations. Affected cattle may present ascites, emaciation, intermittent dark diarrhea, tenesmus, rectal prolapse, and neurological signs resulting from hepatic encephalopathy. Hepatogenous photosensitization may be observed but is uncommon. Necropsy findings include dependent subcutaneous edema, ascites, and edema of the mesentery, abomasal folds, and gallbladder. The liver is firm, normal or reduced in size with a white, thick capsule. Microscopically, the hallmarks of Senecio-associated disease are varying degrees of hepatocellular megalocytosis, bile duct hyperplasia and fibrosis. Sheep are significantly more resistant to Senecio intoxication than are cattle and avidly ingest Senecio plants; therefore, the use of sheep for grazing infested pasture is recommended for preventing the associated disease in cattle.

Assessment of cyclohexanone toxicity in inhalation-exposed F344 rats and B6C3F1 mice: applications in occupational health

Background: Cyclohexanone is a chemical used in various industries and many workers are exposed to it. Therefore, in this study, we determined the toxicity of cyclohexanone in inhalation-exposed F344 rats and B6C3F1 mice, so as to apply the findings in hazard and risk assessments.Methods: Ten male and 10 female rats and mice per test group were exposed to cyclohexanone vapors at 0, 100, 250, and 625 ppm concentrations for 6 h per day, 5 d per week, and for 13 weeks. All rats and mice were killed after the exposure period. Clinical signs, body weight, feed intake, and ophthalmoscopy findings were recorded during the exposure period, and hematology, blood biochemistry, organ weights, gross findings, and histopathology were evaluated thereafter.Results: The following findings were noted in cyclohexanone-exposed F344 rats: increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, increased liver weight, and bile duct hyperplasia in the males exposed to 250 and 625 ppm cyclohexanone, increased ALT levels and bile duct hyperplasia in the females exposed to 625 ppm cyclohexanone, and increased blood urea nitrogen (BUN) and tubular basophilia in the renal cortex in the males exposed to 625 ppm cyclohexanone. On the other hand, B6C3F1 mice exposed to cyclohexanone showed no obvious exposure-related effects.Conclusion: Based on the findings, the no-observed-adverse-effect level (NOAEL) was determined to be 100 ppm in F344 rats and >625 ppm in B6C3F1 mice. Therefore, 2 ppm was revealed as the derived no-effect level (DNEL) for cyclohexanone.