Hepatobiliary Disease Research Articles

An epidemiological surveillance study (2021–2022): detection of a high diversity of Clostridioides difficile isolates in one tertiary hospital in Chongqing, Southwest China

BackgroundClostridioides difficile is a bacterium that causes antibiotic-associated infectious diarrhea and pseudomembranous enterocolitis. The impact of C. difficile infection (CDI) in China has gained significant attention in recent years. However, little epidemiological data are available from Chongqing, a city located in Southwest China. This study aimed to investigate the epidemiological pattern of CDI and explore the drug resistance of C. difficile isolates in Chongqing.MethodsA case-control study was conducted to investigate the clinical infection characteristics and susceptibility factors of C. difficile. The features of the C. difficile isolates were evaluated by testing for toxin genes and using multi-locus sequence typing (MLST). The susceptibility of strains to nine antibiotics was determined using agar dilution technique.ResultsOut of 2084 diarrhea patients, 90 were tested positive for the isolation of toxigenic C. difficile strains, resulting in a CDI prevalence rate of 4.32%. Tetracycline, cephalosporins, hepatobiliary disease, and gastrointestinal disorders were identified as independent risk factors for CDI incidence. The 90 strains were classified into 21 sequence types (ST), with ST3 being the most frequent (n = 25, 27.78%), followed by ST2 (n = 10, 11.11%) and ST37 (n = 9, 10%). Three different toxin types were identified: 69 (76.67%) were A+B+CDT−, 12 (13.33%) were A−B+CDT−, and 9 (10%) were A+B+CDT+. Although substantial resistance to erythromycin (73.33%), moxifloxacin (62.22%), and clindamycin (82.22%), none of the isolates exhibited resistance to vancomycin, tigecycline, or metronidazole. Furthermore, different toxin types displayed varying anti-microbial characteristics.ConclusionsThe strains identified in Chongqing, Southwest China, exhibited high genetic diversity. Enhance full awareness of high-risk patients with HA-CDI infection, particularly those with gastrointestinal and hepatocellular diseases, and emphasize caution in the use of tetracycline and capecitabine. These findings suggest that a potential epidemic of CDI may occur in the future, emphasizing the need for timely monitoring.

Prevention of Portal-Tract Fibrosis in Zfyve19-/- Mouse Model with Adeno-Associated Virus Vector Delivering ZFYVE19.

Zinc finger FYVE-type containing 19 (ZFYVE19) deficiency, caused by biallelic ZFYVE19 complete loss-of-function variants, is a recently identified chronic hepatobiliary disorder characterized by obvious portal-tract fibrosis, increased numbers of bile ducts with malformations, and abnormal levels of serum markers of hepatobiliary injury. As liver-targeted adeno-associated virus (AAV) gene therapy has been used successfully in hepatobiliary diseases, liver-targeted gene therapy has been explored in a mouse model of this disorder. Three ZFYVE19 AAV vectors (AAV-hZFYVE19, AAV-hZFYVE19-m, and AAV-hZFYVE19-co) were constructed and injected into Zfyve19-/- mice, which were treated with alpha-naphthyl isothiocyanate, a hepatobiliary toxin. Hematoxylin/eosin, immunohistochemical staining, immunofluorescence staining, Sirius Red staining, real-time quantitative PCR, and Western blotting of liver tissue, along with serum hepatobiliary injury marker analyses, were performed to evaluate the effects of gene therapy. AAV-hZFYVE19 decreased serum hepatobiliary injury markers, portal-tract inflammation, ductal hyperplasia, and portal-tract fibrosis in the Zfyve19-/- model mice most substantially at a relatively low dose (1 × 1011 vg/kg), whereas AAV-hZFYVE19 at a higher dose gradually lost the abovementioned benefits and even caused deterioration at the highest dose of 5 × 1012 vg/kg. These observations verified the pathogenicity of ZFYVE19 deficiency and suggested that the ZFYVE19 gene needs to function well at an optimal level of expression; both too low and too high a ZFYVE19 expression may be harmful.

Simultaneous determination of seven bile acids to study the effect of ivermectin on their plasma levels in rat by UHPLC–MS/MS

Bile acids (BAs) are considered to be important diagnostic biomarkers to understand the pathophysiology of hepatobiliary and metabolic diseases. BAs regulate lipid and glucose metabolism by binding to farnesoid X receptor (FXR). To date, there were no reports on the effect of an exogenous FXR modulator, ivermectin (IVM), on the plasma BA profiles in rats. To explore the effect of IVM on plasma BA levels in rat, an ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC–MS/MS) method was developed and validated for simultaneous determination of seven major BAs in rat plasma. The developed method was selective, specific, accurate and precise for the quantification of plasma BAs. Sprague–Dawley rats were orally administered with IVM at a dose of 5 mg/kg once a day for 14 days and the plasma BAs were determined before and after IVM exposure using developed UHPLC–MS/MS method. Once-daily administration of IVM for 14 days resulted in significant reduction in cholic acid and deoxycholic acid levels while glycodeoxycholic acid and taurodeoxycholic acid levels were not affected. Interestingly, tauro-α-muricholic acid and tauro-β-muricholic acid levels were significantly increased. This study revealed that IVM has an important effect on plasma BA profiles in rats. This report provides an analytical methodology that can be applied to investigate the effect of drugs or pathophysiological factors on plasma BA levels.

Hepatobiliary disease after bone marrow transplant: A cross-sectional study of 377 patients.

Bone marrow transplantation (BMT) is a standard treatment for several haematologic conditions. Following BMT, patients may develop hepatobiliary complications that impact morbidity and mortality. The differential diagnosis may include drug-induced liver injury (DILI), sepsis-associated liver injury (SALI), sinusoidal obstruction syndrome (SOS), graft-versus-host disease (GVHD), viral hepatitis, ischaemic hepatitis, and fulminant hepatitis. To evaluate the frequency, clinical characteristics, and outcomes of patients with hepatobiliary alterations associated with BMT in a tertiary referral centre. This was a cross-sectional study with data collected from the medical records of patients undergoing BMT between January 2017 and June 2022. We diagnosed hepatobiliary complications based on established criteria. We included 377 patients; 55.7% had hepatobiliary complications. Female gender, pre-BMT hepatobiliary alteration, and haploidentical allogeneic transplantation were associated with increased risk with odds ratios (OR) of 1.8 (p = 0.005), 1.72 (p = 0.013) and 3.25 (p = 0.003), respectively. Patients with hepatobiliary complications spent longer in the hospital than those without (27.7 × 19.3 days, respectively; p < 0.001). Among 210 patients with hepatobiliary complications, 28 died compared to 5 of 167 without complications (OR 4.98; p = 0.001). Hepatobiliary complications are frequent in patients undergoing BMT. There is a greater risk of their occurrence in women, people with pre-BMT liver alterations, and in haploidentical transplants. The occurrence of these complications increases the length of stay and is associated with a higher risk of death.

Intestinal Microbiome in Dogs with Chronic Hepatobiliary Disease: Can We Talk about the Gut-Liver Axis?

The gut-liver axis represents a current topic in human medicine. Extensive research investigates the gut microbiome (GM) modifications in relation to various kinds of chronic hepatobiliary diseases (CHD), with many mechanisms and therapeutical implications recognized. Those aspects in veterinary medicine are still quite unexplored. The aim of the present study was to evaluate GM in dogs diagnosed with CD. Comparison among CHD dogs were made considering some clinical and biochemical variables (lipemia and alanine-aminotransferase activities), presence of cholestasis or endocrine disorders, diet). Sixty-five dogs were prospectively enrolled with clinical and hematobiochemical evaluation and 16S-RNA GM sequencing assessed. Dogs that received antibiotics and/or pre/pro/symbiotics administration were excluded. Deeper GM alteration was observed between dogs with or without ultrasonographic and biochemical cholestatic CHD. Cholestasis was associated with a decrease in several bacterial taxa, including Clostridium hiranonis, Fusobacterium, Megamonas, Ruminococcus faecis, Turicibacter, and higher levels of Escherichia/Shigella and Serratia. Thus, the alteration in bile flow and composition, typical of cholestasis, may directly affect the local intestinal microbial environment. For the management of dogs with CHD and especially cholestatic CHD, clinicians should be aware that gut-liver interaction may lead to dysbiosis.

Adverse Reaction Signals Mining of Vonoprazan: A Pharmacovigilance Study Based on FAERS

Background. Vonoprazan is a novel selective and noncompetitive oxidative-reduction state proton pump inhibitor (PPI), known as a potassium-competitive acid blocker (P-CAB). Due to its novelty, the clinical application of vonoprazan is relatively limited, and its long-term safety and adverse effects still need to be further studied and confirmed. Methods. We used the data from the FDA adverse eventreporting system (FAERS) from 2015 to 2022. Disproportionality method including calculation of the reporting odds ratio (ROR) and calculation of the information component (IC) was used to detect potential adverse drug reactions (ADRs). In preferred terms, the significant signals are ranked in the following order: Plateletcrit increased (ROR 1447.3 IC 36.62), benign duodenal neoplasm (ROR 11157.84 IC 36.40), gallbladder volvulus (ROR 964.77 IC 36.20), myopathy endocrine (ROR 723.58 IC 35.88), pernio-like erythema (ROR 723.58 IC 35.88), septic coagulopathy (ROR 723.58 IC 35.88), and so on. In SOCs, the significant signals are ranked in the following order: hepatobiliary disorders (ROR 5.65 IC 29.15), metabolism and nutrition disorders (ROR 2.78 IC 28.12), blood and lymphatic system disorders (ROR 2.73 IC 28.12), investigations (ROR 2.19 IC 27.76), endocrine disorders (ROR 2.09 IC27.76), gastrointestinal disorders (ROR 1.87 IC 27.52), and so on. Conclusion. Despite its potential advantages, there is still limited clinical experience with vonoprazan, and the long-term safety and adverse effects of this drug are not fully understood. Further studies are needed to confirm its safety and efficacy in a larger patient population and to establish its role in the management of acid-related disorders. In the meantime, careful monitoring and patient education are recommended for those who are prescribed vonoprazan.

Gemcitabine with Cisplatin Versus Hepatic Arterial Infusion Pump Chemotherapy for Liver-Confined Unresectable Intrahepatic Cholangiocarcinoma

BackgroundA post-hoc analysis of ABC trials included 34 patients with liver-confined unresectable intrahepatic cholangiocarcinoma (iCCA) who received systemic chemotherapy with gemcitabine and cisplatin (gem-cis). The median overall survival (OS) was 16.7 months and the 3-year OS was 2.8%. The aim of this study was to compare patients treated with systemic gem-cis versus hepatic arterial infusion pump (HAIP) chemotherapy for liver-confined unresectable iCCA.MethodsWe retrospectively collected consecutive patients with liver-confined unresectable iCCA who received gem-cis in two centers in the Netherlands to compare with consecutive patients who received HAIP chemotherapy with or without systemic chemotherapy in Memorial Sloan Kettering Cancer Center.ResultsIn total, 268 patients with liver-confined unresectable iCCA were included; 76 received gem-cis and 192 received HAIP chemotherapy. In the gem-cis group 42 patients (55.3%) had multifocal disease compared with 141 patients (73.4%) in the HAIP group (p = 0.023). Median OS for gem-cis was 11.8 months versus 27.7 months for HAIP chemotherapy (p < 0.001). OS at 3 years was 3.5% (95% confidence interval [CI] 0.0–13.6%) in the gem-cis group versus 34.3% (95% CI 28.1–41.8%) in the HAIP chemotherapy group. After adjusting for male gender, performance status, baseline hepatobiliary disease, and multifocal disease, the hazard ratio (HR) for HAIP chemotherapy was 0.27 (95% CI 0.19–0.39).ConclusionsThis study confirmed the results from the ABC trials that survival beyond 3 years is rare for patients with liver-confined unresectable iCCA treated with palliative gem-cis alone. With HAIP chemotherapy, one in three patients was alive at 3 years.

THU471 Resolution Of Elevated Alkaline Phosphatase After Joint Replacement In Severe Osteoarthritis

Abstract Disclosure: J. Rakholiya: None. S. Nagaraj: None. P. Sharma: None. Introduction: Alkaline phosphatase (ALP) elevation is commonly seen in hepatobiliary disorders. Conditions of high turnover such as healing fractures, hyperparathyroidism, Pagets’ disease of the bone, also lead to ALP elevation (1). Osteoarthritis (OA), characterised by degeneration of articular cartilage, joint space narrowing, and osteophyte formation, rarely causes ALP elevation. We present a case of ALP elevation that resolved after the arthritic joint was replaced. Case Presentation: A 60-year-old female with history of bilateral knee OA presented to the endocrinology clinic due to elevated ALP for over a year. She reported chronic fatigue, sleep disturbance, dry skin, hair loss, inability to lose weight and anxiety. ALP level at presentation was 165 U/L. An extensive work-up showed unremarkable serum and ionized calcium, serum phosphorus, parathyroid hormone, vitamin D 25-OH, calcitriol, 24-hour urine calcium, serum protein electrophoresis, and 24-hour urine protein electrophoresis. Dual x-ray absorptiometry (DXA) scan was normal with Fracture risk assessment tool (FRAX) score 5.9% for major osteoporotic fracture and 0.2% for hip fracture. Patient underwent total left knee arthroplasty following which ALP improved from 165 U/L to 135 U/L within 9 weeks of the procedure. Five months later, ALP was elevated (164 U/L) again. She then underwent right knee arthroplasty and ALP dropped to normal level (114 U/L) within 1 week post-surgery. Discussion: We report the rare phenomenon of ALP elevation in severe OA, that resolved after joint replacement. OA is characterised by inflammation mediated cartilage destruction, subchondral bone change, osteophyte formation, and alterations of ligaments and meniscuses. Bone change in severe OA is associated with increased synovial fluid ALP level. However, there is a paucity of literature studying the association of ALP with OA. A cross-sectional study of 3060 patients demonstrated higher serum ALP with symptomatic knee OA. Another study showed higher ALP levels with increased joint stiffness. The ALP elevation was attributed to low grade inflammation causing an inflammatory response in chondrocytes, and osteoarthritic pain. There is a need for more studies to evaluate the utility of serum ALP as a biomarker for severe OA. Presentation: Thursday, June 15, 2023

Role of Neutrophil to Lymphocyte Ratio (NLR) and C-Reactive Protein to Albumin Ratio (CAR) as Early Predictors of Severity in Acute Pancreatitis

Background: Acute pancreatitis (AP) is a life-threatening disease caused by a variety of factors, and once it progresses to severe acute pancreatitis, the prognosis is poor. Different modalities are available for predicting severity in acute pancreatitis. A single, cheap, widely available marker with high sensitivity and specificity is yet to be identified. Aims and objectives: The present study was aimed to determine the relation of the neutrophil-lymphocyte ratio (NLR) and CRP-Albumin ratio (CAR) in early prediction of severity in acute pancreatitis. Materials and methods: This cross-sectional study was conducted at the Department of Gastrointestinal, Hepatobiliary and Pancreatic Disorders (GHPD), BIRDEM General Hospital, Shahbagh, Dhaka, Bangladesh, from April, 2020 to March, 2022. Diagnosis of acute pancreatitis was made by clinical findings, serum amylase and lipase levels (&gt;3 times the upper limit of normal values), evidences of acute pancreatitis by ultrasonography and computed tomography (CT). Severity of acute pancreatitis was classified according to the revised version of Atlanta classification. Data collection was done through a structured questionnaire. Data were analyzed by SPSS 23. Receiver operating characteristic (ROC) curve was constructed to estimate the sensitivity and specificity of NLR and CAR. Results: A total of 120 patients with acute pancreatitis were enrolled in this study. Age of the patients was 45.20±13.93 (mean ±SD) years, male predominance was observed (54.2%). Majority of the cases were mild 52.5% (n=63) compared to moderate 28.3% (n=34) and severe 19.2% (n=23). The NLR was 6.63±3.344 (Mean ± SD) with a range of 1.32 to 18.75. The CAR was 3.20±2.23 (Mean ±SD) with range of 0.14-7.20. The area under the curve (AUC) of NLR and CAR were 0.865 and 0.949 for severity of AP respectively. Sensitivity, specificity, PPV, NPV and accuracy of NLR at cut-off ≥5.72 were 80.7%, 76.2%, 75.4%, 81.4% &amp; 78.3% and that of CAR at cut-off ≥2.07 were 96.5%, 76.2%, 78.6%, 96.0% &amp; 85.8%. Conclusion: This study revealed that NLR and CAR are good predictors in the assessment of severity of AP. These easily accessible and low-cost inflammatory markers can be used for the management of acute pancreatitis. Bangladesh Crit Care J September 2023; 11 (2): 83-89

Frequency, Clinical Presentation, and Outcome of Acute-on-Chronic Liver Failure among Decompensated Cirrhosis of Liver Patients in a Tertiary Care Hospital

Background: Acute-on-chronic liver failure (ACLF) is characterised by the presence of organ failure in patients with decompensated cirrhosis and is associated with high short-term mortality. Different international entities have taken initiatives to define the condition in different times but recommendations and definitions from The European Association for the Study of the Liver- Chronic Liver Failure (EASL-CLIF) Consortium Acute-on-Chronic Liver Failure in Cirrhosis (CANONIC) study are most comprehensive and widely accepted till date. Only limited data are available on the prevalence, clinical characteristics, and short-term outcomes of ACLF in Bangladesh. It would be very useful for clinicians to identify patients with ACLF early and initiate focused therapy including referral to transplant centers if these data are available. Objective: To evaluate frequency, clinical presentation, and outcome of acute-on-chronic liver failure among decompensated cirrhosis of liver patients. Materials and Methods: This prospective observational study was carried out at the Department of Gastrointestinal, Hepatobiliary and Pancreatic Disorders (GHPD), BIRDEM General Hospital, Shahbagh, Dhaka, Bangladesh from July, 2019 to September, 2021. Total 175 patients with decompensated cirrhosis of liver were screened, out of which 22 patients were dropped out due to various reasons. Purposive type of non-probability sampling technique was used. Formal ethical clearance was taken from the Institutional Review Board and ethical measures were ensured in concordance with the Declaration of Helsinki. An informed written consent was taken from all participants. Diagnosis of decompensated cirrhosis was based on clinical, biochemical, radiological and endoscopic findings. Laboratory data sent within 24 hours were collected. Oxygen saturation was measured using fingertip pulse oximeter. Investigations for ACLF triggers were done as necessary which included but not limited to urine routine and microscopic examination, urine culture, blood culture, and Anti HEV IgM. Patients’ prognosis and survivability were observed by follow up phone call at 30 days. All data were recorded in a separate case record form and finally, it was analyzed by SPSS 23. Results: Out of 153 patients, 49 patients (32%) had ACLF: grade 1 ACLF in 26 (17%), grade 2 in 18 (11.8%), and grade 3 in 5 (3.3%) patients. Patients had an average age of 59.54±11.55 years with no significant difference between ACLF and no ACLF groups. Most patients in both groups had others (NAFLD, autoimmune hepatitis, secondary biliary cirrhosis, idiopathic) as the main underlying cause of cirrhosis. Bacterial infection, GI bleeding, HEV infection, reactivation of HBV were the precipitating events in 81.6% of patients with ACLF, with bacterial infection being the most common trigger (63.3%). Overall, 44.9% ACLF patients died within 30 days of admission. Older age, male sex, hepatic encephalopathy, GI bleeding, presence of any trigger and higher Child-Turcotte-Pugh (CTP) score were associated with increased risk of death in ACLF. Conclusion: Follow up of 153 patients with decompensated cirrhosis of liver revealed that 1 in 3 patients had ACLF and 44% of them would die in 30 days. Bacterial infection and GI bleeding were the most common triggers of ACLF. Early identification and intervention with multidisciplinary approach and referral to transplant centers are likely to improve survival outcomes in this population. Bangladesh Crit Care J September 2023; 11 (2): 113-120

Liver Abscess Caused by Klebsiella Pneumoniae, Series of Cases with Different Management

Abstract We present two cases of liver abscess caused by Klebsiella pneumoniae with different management, one of them resolved under antibiotherapy without theraputic drainage. Klebsiella pneumoniae is a gram-negative, lactose-fermenting, rod-shaped bacterium that is part of natural intestinal flora and one of the common causes of nosocomial infections. The liver is one of the viscera in which abscesses frequently occur and is associated with intraabdominal or hepatobiliary diseases. The first patient was admitted to „Prof. Dr. Agrippa Ionescu” Hospital with sepsis due to liver abcess. The CT examination revealed a liver lesion in segments VI-VII, limited by a hypodense margin with fluid-parafluid content (with maximal axial diameters 54/32 mm), without any other focal hepatic lesions. Laboratory workup revealed neutrophilia and lymphopenia, anemia, trombocytopenia, increased inflammatory syndrome, a high level of procalcitonin and blood cultures positives with Klebsiella pneumoniae sensitive. Based on the information presented, the diagnosis established was liver abscess caused by Klebsiella pneumoniae with favorable outcome under initial empirical antibiotherapy with Vancomycin and Meropenem, de-escalated later to I.V. Ceftriaxone, due to the result of the antibiogram. At the 14 day mark, an abdominal CT reveals a slight dimentional progression. Percutaneous ecographic guided drainage was considered but the patient denied it.The patient was released with the antibiotherapy recomandation of 14 more days of orally administered Cefixime. After 4 weeks of antibiotherapy treatment, the patient is ecographically reassesed. The liver abscess is observed to have achieved complete remission without drainage, despite the classical literature recommended protocol. The second case, a male patient admitted with sepsis due to multiple liver abscess (the largest one 100/50 mm with negative blood cultures and unfavorable outcome, despite maximal empirical therapy. Percutaneous ecographic guided drainage was performed and Klebsiella pneumoniae was isolated from the drainage samples .Under target antibiotherapy with Ertapenem for 4 weeks the outcome was very good with complete resolution of the hepatic lesions. Hepatic abscess is associated with a high mortality rate and several complications, despite its low incidence. Prompt recognition is important for effective management and achieving good outcomes. Usually, the liver abscess is treated by either percutaneous drainage or surgical drainage in combination with antibiotics.The key to succesful outcomes is early diagnosis, institution of appropriate therapy and attentive evolution monitoring.More prospective trials with large cohorts are needed to refine our understanding of this serious condition.

SARS-COV 2 pandemisi sonrası Hatay (Türkiye) ilinde kedilerin son sağlık durumu nedir? ikinci bölüm

The current study, as episode two, focused on the latest status of gastrointestinal, urinary tract, and hepatobiliary diseases, which might be induced by stress in cats following the SARS-CoV-2 pandemic in Hatay province in Türkiye. The study looked at post-pandemic (June 2021–June 2022) diagnosis data of cats with gastrointestinal, urinary tract, and hepatobiliary diseases (n: 147), admitted to Hatay Mustafa Kemal University Veterinary Health, Practice, and Research Center. The average age of the cats admitted to the clinic was 33.04 ± 2.77 months in post-pandemic period. The ages of the cats diagnosed with gastrointestinal, urinary tract and hepatobiliary diseases were 27.78 ± 4.33, 38.04 ± 3.95, and 40.12 ± 6.27, respectively. There were no breed and sex difference for diseases diagnosed in post-pandemic duration. Distributions of gastrointestinal, urinary, and hepatobiliary diseases were determined as 51.0%, 37.4%, and 11.6%, respectively. There was no statistical difference between the cat breeds that presented to the clinic in terms of gastrointestinal, urinary, and hepatobiliary diseases. Gastroenteritis (40%), gastritis (17.33%), and unconfirmed and suspected Feline inflammatory bowel disease (13.33%) are more diagnosed digestive system diseases. Cystitis (54.54%) and hemorrhagic cystitis (32.72%) were commonly diagnosed in urinary diseases. Besides, cholangiohepatitis was commonly diagnosed either alone (41.17%) or accompanied by hepatic lipidosis (23.52%) or cholecystitis (17.65%) in cats following the pandemic. Consequentially, although restrictions implemented during the pandemic that caused stress on cats were ended, the chronic impacts of stress seem to be continuing on cats, especially in terms of the gastrointestinal system, following the pandemic.

Covert dissemination of pLVPK-like virulence plasmid in ST29-K54 Klebsiella pneumoniae: emergence of low virulence phenotype strains.

This study aimed to explore the epidemic, clinical characteristics, and molecular and virulence attributes of Klebsiella pneumoniae serotype K54 (K54-Kp). A retrospective study was conducted on 328 strains of Klebsiella pneumoniae screened in a Chinese hospital from January 2016 to December 2019. The virulence genes and antibiotic resistance genes (ARGs) were detected by PCR, and a drug sensitivity test was adopted to detect drug resistance. Multilocus sequence typing (MLST) and PFGE were performed to determine the clonal correlation between isolates. Biofilm formation assay, serum complement-mediated killing, and Galleria mellonella infection were used to characterize the virulence potential. Our results showed that thirty strains of K54-Kp were screened from 328 strains of bacteria, with an annual detection rate of 2.29%. K54-Kp had a high resistance rate to antibiotics commonly used in the clinic, and patients with hepatobiliary diseases were prone to K54-Kp infection. MLST typing showed 10 sequence typing, mainly ST29 (11/30), which concentrated in the B2 cluster. K54-Kp primarily carried virulence genes of aerobactin, silS, allS, wcaG, wabG, and mrkD, among which the terW gene was closely related to ST29 (p<0.05). The strains infected by the bloodstream had strong biofilm formation ability (p<0.05). Most strains were sensitive to serum. Still, the virulence of pLVPK-like virulence plasmid in ST29-K54 Klebsiella pneumoniae was lower than that of ST11 type and NTUH-K2044 in the Galleria mellonella model. Therefore, these findings supply a foundation to roundly comprehend K54-Kp, and clinicians should strengthen supervision and attention.

B-118 Evaluation of Thirteen Commercially Available Clinical Chemistry Assays on the Alinity® c System

Abstract Background Expansion of instrument analyte menus is often needed to accommodate country specific needs. To address specific needs in China, thirteen clinical chemistry assays have been modified for evaluation on the Alinity c system. These analytes are used in the investigation of several diseases including liver disease, hepatobiliary disease, cardiovascular and cerebrovascular disease, kidney disease, inflammation, infectious disease, alcoholism and glucose metabolism disease. Methods The following analytes were evaluated: a1-Microglobulin (a1-MG), Ischemia modified albumin (IMA), Glutamate Dehydrogenase (GLDH), Fibrin Degradation Products (FDP), Myeloperoxidase (MPO), Fibronectin (FN), Pyruvate (PYR), Monoamine Oxidase (MAO), Heart-type Fatty Acid-Binding Protein (H-FABP), Cholyglycine (CG), Serum Amyloid A Protein (SAA), Ethanol (EtOH) and Ferritin (Fer). Reagents from Beijing Strong Biotechnologies Inc. (BSBE) were evaluated on the ABBOTT Alinity c system. Precision, LoQ and linearity range were evaluated with guidance from CLSI documents EP15-A3, EP17-A2 and EP06-Ed2. Correlation to commercially available assays were performed using human serum/plasma/urine samples across the measuring range. Results The table below summarizes key performance of assays. Conclusions These initial results are very promising. All the assays tested exhibit good precision, linearity, anti-interference, and LOQ performances on Alinity c system. In addition, they also have excellent correlation with Jinya assays from BSBE on ARCHITECT system except SAA assay, and SAA has excellent correlation with Siemens BNII SAA assay.

Quality uniformity evaluation of Babao Dan capsule based on its 14 main chemical compounds and in vitro anti-inflammatory activity

Babao Dan capsule (BBDC) is a state secrecy traditional Chinese medicine patent prescription (TCMPP) that is widely used for hepatobiliary and urinary diseases. However, its quality uniformity evaluation (QUE) is lacking. It is necessary to develop a novel and multi–dimensional QUE strategy for BBDC based on chemical properties and bioactivities. An ultrahigh performance liquid chromatography–triple quadrupole–mass spectrometry (UHPLC–QQQ–MS/MS) was developed to quantify 14 compounds, and a QUE strategy was to assess intra–batch and inter–batches consistency. Furthermore, the anti-inflammatory activities were measured in vitro by RAW264.7 cell and NF-κB-RE-luc HEK293 cell. The results indicated that 14 compounds from 16 samples (10 batches) were accurately quantified. And the intra–batch consistency values were less than or equal to 20.1%, whereas the largest inter–batches consistency value was 167.5% of glycocholic acid. 10 batches were divided into three categories to distinction inter–batches differences. BBDC had anti-inflammatory activities including inhibition of LPS–induced NO production and TNF α–activated luciferase counts. The NO inhibition difference of 10 batches was 10.87 μM and accounted for 49.8%. Briefly, the anti-inflammatory activities of 10 batches were different, but the dynamic trends within the same batch were similar, which was supported by luciferase counts. And the correlation was confirmed that the main compounds related to anti-inflammatory in BBDC were bile acids and saponins. Hence, a QUE strategy for BBDC was proposed to improve quality and ensure efficacy. This strategy can be utilized as a reference for consistency evaluations of TCMPPs.

Evaluation of Gastrointestinal System Complaints and Comorbidities in Pediatric Familial Mediterranean Fever Patients.

Familial Mediterranean fever (FMF) is the most prevalent hereditary autoinflammatory disease among children. Abdominal pain and various gastrointestinal system (GIS) manifestations may arise directly from FMF or concomitantly with FMF. This study aimed to evaluate GIS complaints and findings other than classic peritonitis attacks in patients with FMF and to interpret concomitant GIS and hepatic disorders in these patients. The medical and genetic findings of patients with FMF who attended our clinic between December 2011 and December 2021 were reviewed. Gastrointestinal system symptoms, liver function tests, abdominal images, and endoscopic and histopathological data were extracted from medical records. A total of 576 pediatric patients (female, 52.3%) diagnosed with FMF were included. Among them, almost one-fifth displayed GIS complaints, such as abdominal pain, defecation problems, and dyspepsia, distinct from typical FMF attacks. High serum aminotransferase levels were detected in 18.4% of the patients, with viral infections being the most common cause of moderate/severe hypertransaminasemia. In addition, during follow-up, 26.9% of them were referred to the pediatric gastroenterology department. At least 1 gastroenterological and hepatobiliary disorder was detected in 17.5% of the patients because of organic and functional GIS disorders or hepatobiliary disorders, such as gastroesophageal reflux disease, esophagitis, functional dyspepsia, and inflammatory bowel diseases. Various GIS and hepatic disorders can be encountered in children with FMF. The spectrum of these complaints and pathologies can range from frequently observed health problems to more severe diseases.

Loss of Mtm1 causes cholestatic liver disease in a model of X-linked myotubular myopathy.

X-linked myotubular myopathy (XLMTM) is a fatal congenital disorder caused by mutations in the MTM1 gene. Currently, there are no approved treatments, although AAV8-mediated gene transfer therapy has shown promise in animal models and preliminarily in patients. However, 4 patients with XLMTM treated with gene therapy have died from progressive liver failure, and hepatobiliary disease has now been recognized more broadly in association with XLMTM. In an attempt to understand whether loss of MTM1 itself is associated with liver pathology, we have characterized what we believe to be a novel liver phenotype in a zebrafish model of this disease. Specifically, we found that loss-of-function mutations in mtm1 led to severe liver abnormalities including impaired bile flux, structural abnormalities of the bile canaliculus, and improper endosome-mediated trafficking of canalicular transporters. Using a reporter-tagged Mtm1 zebrafish line, we established localization of Mtm1 in the liver in association with Rab11, a marker of recycling endosomes, and canalicular transport proteins and demonstrated that hepatocyte-specific reexpression of Mtm1 could rescue the cholestatic phenotype. Last, we completed a targeted chemical screen and found that Dynasore, a dynamin-2 inhibitor, was able to partially restore bile flow and transporter localization to the canalicular membrane. In summary, we demonstrate, for the first time to our knowledge, liver abnormalities that were directly caused by MTM1 mutation in a preclinical model, thus establishing the critical framework for better understanding and comprehensive treatment of the human disease.

Microbiome analysis of bile from apparently healthy cats and cats with suspected hepatobiliary disease.

Bacterial infection of bile is a common cause of hepatobiliary disease in cats. Whether bile harbors a core microbiota in health or in cases of suspected hepatobiliary disease in cats is unknown. Establish if gallbladder bile in apparently healthy cats harbors a core microbiota composed of bacterial taxa common to many individuals. Compare results of bile cytology, bile culture, and 16S rRNA gene amplicon sequencing in apparently healthy cats and cats with suspected hepatobiliary disease. Forty-three client-owned cats with suspected hepatobiliary disease and 17 control cats. Bile was collected by ultrasound guided cholecystocentesis (cats with suspected hepatobiliary disease) or laparotomy after euthanasia (controls). Bile samples underwent cytologic examination, aerobic and anaerobic culture, and DNA was extracted for 16S rRNA gene amplification and sequencing. Microbiome sequencing did not identify a core microbiota in control cats or cats having bile sampled because of clinical suspicion for hepatobiliary disease. Microbiome profiles from control cats were indistinguishable from profiles obtained from sampling instruments and reagents that were not exposed to bile (technical controls). Bacterial taxa that could not be explained by contamination or off-target amplification were identified only in samples from cats with bactibilia and positive bile culture results for Escherichia coli. In several E. coli positive samples, microbiome sequencing also identified a small number of potentially co-infecting bacterial genera not identified by culture. Cat bile does not harbor a core microbiota. Uncultured bacteria may contribute to pathogenesis of hepatobiliary disease in cats with bile E. coli infection.

O-Fenilendiamin ve Benzokinon Bazlı Polimer Filmle Oluşturulan Kolesterol Biyosensörünün Elektrokimyasal Karakterizasyonu

Aim: The detection of cholesterol concentration in the blood is an important parameter in the diagnosis and follow-up of many diseases, including cardiovascular and hepatobiliary diseases. Nowadays, various methods have been used for the routine analysis of cholesterol, including spectrophotometric, high-performance liquid chromatography (HPLC), and amperometric methods. Since biosensors have advantages such as high sensitivity, fast response, low cost, small size, continuous online detection, and reproducible results, it is aimed to develop a biosensor method for cholesterol determination. Methods: Polymer film immobilization was performed on the gold electrode surface electrochemically in an acetonitrile-water medium containing o-phenylenediamine and benzoquinone. Cholesterol oxidase (ChOx, E.C. 1.1.3.6) enzyme was immobilized on the polymer film-immobilized electrode surface by the self-forming monolayer method. Optimization and characterization studies were carried out for the determination of cholesterol with the prepared electrodes. Results: The determination of cholesterol was performed via monitoring the oxidation current of enzymatically produced H2O2 at 0.70 V Ag/AgCl. Optimum biosensor conditions were determined for phosphate buffer at 0.1M, pH 7.0, and 30°C for the Au/oPD-BQ/ChOx enzyme electrode. The linear working range is 9.8×10-6-1.1×10-2 mM, and the cholesterol detection limit is 9.8×10-6 mM. The analysis of total cholesterol in solutions was performed using the proposed Au/oPD-BQ/ChOx electrode. Conclusion: A convenient, simple, reproducible, and stable method was developed for the determination of cholesterol by immobilizing cholesterol oxidase on the prepared electrode surface through a polymer film. The sensor we designed can be expanded to improve the detection of many analytes in the clinic.

Long-term Plasmapheresis for Cholestatic Pruritus

Cholestatic pruritus is commonly associated with many hepatobiliary disorders and can be typically controlled with conventional treatment. Therapeutic plasma exchange (TPE) has shown success in improving untreatable cholestatic pruritus with great success. We report on the use of TPE to alleviate debilitating pruritus in one patient with a two-year history of chronic pruritus refractory to conventional treatment. The patient presented with an acute onset of diffuse pruritus that did not fully respond to first or second-line treatments, including bile acid sequestrants. Pruritus significantly affected her quality of life, interfering with daily activity, causing insomnia and weight loss. Due to the severity of her symptoms, biweekly TPE with 5% albumin infusions were initially trialed. Patient found symptoms to be much improved until about the fifth day after each TPE. TPEs were scheduled twice per week alternating with once per week. TPE appeared to be an effective treatment for this patient with chronic cholestatic pruritus. For our patient, TPE seemed to provide relief for about 3-5 days after each TPE.