Case report: Gross total resection of a primary fourth ventricular meningioma using the telovelar approach in a dog.

An 11-year-old spayed female Maltese dog presented with a 2-month history of gait alterations, wide-based stance, and chronic vomiting. Neurological examination revealed cerebellovestibular signs, including head tilt, nystagmus, strabismus, intentional tremor, and hypermetric gait. MRI showed a mass with iso- to hypointensity on T1-weighted (T1W) images and heterogeneous hyperintensity on T2-weighted (T2W) images, with marked non-uniform contrast enhancement. The tumor was removed via a telovelar approach without intraoperative complications. Postoperatively, the dog developed non-ambulatory paraparesis with the rigidity of the pelvic limbs but recovered ambulation within 6 days. Preoperative neurological signs progressively improved, and the patient was discharged without complications 10 days after surgery. Histological examination revealed dense spindle cells with an abundant collagen matrix and oval-shaped nucleated cells with small whorls, leading to a diagnosis of transitional meningioma of the fourth ventricle. MRI follow-up at 8 months postoperatively showed no definitive evidence of recurrence. At the final follow-up, 15.4 months postoperatively, mild neurological signs, including a slight head tilt and subtle strabismus, remained, but the rest of the neurological examination was normal. This is the first reported case of a meningioma in the fourth ventricle of a dog successfully removed using the telovelar approach.

Reconstruction of the Quadriceps Extensor Mechanism with a Calcaneal Tendon-Bone Allograft in a Dog with a Resorbed Tibial Tuberosity Fracture.

A non-reducible tibial tuberosity fracture is a rare complication of tibial tuberosity transposition performed during correcting of medial patella luxation (MPL) in dogs. This condition severely disrupts the quadriceps extensor mechanism, leading to significant pelvic limb lameness. An 11-year-old, 1.8 kg spayed female Yorkshire Terrier sustained a comminuted left tibial tuberosity fracture during surgical correction of an MPL. Six months after surgery, the dog was markedly lame and unable to extend the left stifle. Radiographs revealed patella alta and resorption of the fragmented tibial tuberosity. A composite frozen allogeneic calcaneal tendon-bone block was utilized to reconstruct the tibial tuberosity and reattach the patellar ligament. Initial postoperative radiographs confirmed restoration of a normal patellar ligament to patella length ratio (1.42). Both the allogeneic bone used for tibial tuberosity reconstruction and the tendon used to reattach the patellar ligament were successfully integrated. The dog regained satisfactory limb function without recurrence of patella luxation, as reported by the owners 29 months postoperatively. The use of a calcaneal tendon-bone allograft effectively restored the functional integrity of the quadriceps extensor mechanism, providing a viable option for addressing quadriceps insufficiency resulting from the loss of the osseous tibial insertion.

C1 Facetectomy and Ventral Fixation of Occipitoatlantoaxial Complex for Concurrent Congenital Atlanto-Occipital Dislocation and Atlantoaxial Instability in a Toy Poodle.

An 8-month-old, 3.4 kg, castrated male Toy Poodle was referred for progressive tetraparesis and respiratory disorder without a history of trauma. Repeated computed tomography (CT) and magnetic resonance imaging (MRI) with different positions of the neck revealed concurrent atlanto-occipital dislocation (AOD) and atlantoaxial instability (AAI) with spinal cord compression. This case was unique due to its congenital nature and the absence of trauma. The surgical treatment involved precise removal of the C1 vertebra's ventral articular facet, which was compressing on the spinal cord, attributed to its fixed and malaligned position within the atlantooccipital joint. Following facetectomy, the stabilization of the occipital bone to the C2 vertebra was achieved by screws, wire, and polymethyl methacrylate. Two days after surgery, the dog recovered ambulation and showed gradual improvement in gait, despite mild residual ataxia. Postoperative CT and radiographs showed successful decompression of the spinal cord. The screw loosening was confirmed at 114 days, which was managed successfully by extracting the affected screws. Through the 21-month monitoring period, the dog showed a normal gait with a wide-based stance of the pelvic limbs when standing and experienced no pain. This case represents the first report of concurrent congenital AOD and AAI treated with a ventral surgical approach, contributing new insights to the understanding and management of such complex cranio-cervical junction disorders in veterinary neurosurgery.

Surgical Correction of a Sinus Venosus Atrial Septal Defect with Partial Anomalous Pulmonary Venous Connections Using Cardiac Computed Tomography Imaging and a 3D-Printed Model.

Sinus venosus atrial septal defects (SVASDs), concurrent with partial anomalous pulmonary venous connections (PAPVCs), are a rare congenital heart disease in dogs. Surgical correction is essential when clinical signs or significant hemodynamic changes are present. We aimed to report on the successful surgical correction of an SVASD with PAPVCs, using a computed tomography (CT)-based customized 3D cardiac model. A 10-month-old male poodle was referred for corrective surgery for an ASD. Echocardiography confirmed a hemodynamically significant left-to-right shunting flow through an interatrial septal defect and severe right-sided heart volume overload. For a comprehensive diagnosis, a CT scan was performed, which confirmed an SVASD with PAPVCs. A customized 3D cardiac model was used for preoperative decision-making and surgical rehearsal. The defect was repaired using an autologous pericardial patch under a cardiopulmonary bypass (CPB). Temporary pacing was applied for sinus bradycardia and third-degree atrioventricular block. The patient recovered from the anesthesia without further complications. The pacemaker was removed during hospitalization and the patient was discharged without complications 2 weeks post-surgery. At the three-month follow-up, there was no shunting flow in the interatrial septum and the right-sided volume overload had been resolved. The cardiac medications were discontinued, and there were no complications. This report indicates the validity of surgical correction under CPB for an SVASD with PAPVCs, and the advantages of utilizing a CT-based 3D cardiac model for preoperative planning to increase the surgical success rate.

Laparoscopic treatment of paraprostatic cyst in two dogs - complete resection, and partial resection with omentalization: a case report.

Two intact male dogs were evaluated for symptoms, including hematuria, prostatomegaly, anuria, lethargy, and abdominal mass. Presurgical evaluations, including complete physical examinations, blood examinations, abdominal radiography with contrast (only in Case 2), ultrasonography, and computed tomography and magnetic resonance imaging (only in Case 1), were performed. A paraprostatic cyst was diagnosed initially, and laparoscopic exploration and surgery were performed. Complete resection was performed in case 1, whereas partial resection with omentalization was performed in case 2. Histopathological examination of the tissue samples confirmed the presence of paraprostatic pseudocysts in both cases, with no evidence of an epithelial lining. These two cases represent the first documented instances of laparoscopic treatment for extraparenchymal prostatic cysts. The laparoscopic treatment proved feasible even in the case of a giant cyst causing anuria (Case 2). Paraprostatic cysts should be considered a potential differential diagnosis for abnormal urination accompanied by an abdominal mass, and long-term postoperative follow-up is necessary.

Metabolomics and miRNA profiling reveals feature of gallbladder cancer-derived biliary extracellular vesicles.

BACKGROUND: Although there are several studies in the development of various human cancers, the role of exosomes is poorly understood in the progression of gallbladder cancer. This study aims to characterize the metabolic changes occurring in exosomes obtained from patients with gallbladder cancer compared with those from other gallbladder disease groups. METHODS: Biliary exosomes were isolated from healthy donors (n = 3) and from patients with gallbladder cancer (n = 3), gallbladder polyps (n = 4), or cholecystitis (n = 3) using a validated exosome isolation kit. Afterward, we performed miRNA profiling and untargeted metabolomic analysis of the exosomes. The results were validated by integrating the results of the miRNA and metabolomic analyses. RESULTS: The gallbladder cancer group exhibited a significant reduction in the levels of multiple unsaturated phosphatidylethanolamines and phosphatidylcholines compared to the normal group, which resulted in the loss of exosome membrane integrity. Additionally, the gallbladder cancer group demonstrated significant overexpression of miR-181c and palmitic acid, and decreased levels of conjugated deoxycholic acid, all of which are strongly associated with the activation of the PI3K/AKT pathway. CONCLUSIONS: Our findings demonstrate that the contents of exosomes are disease-specific, particularly in gallbladder cancer, and that altered metabolites convey critical information regarding their phenotype. We believe that our metabolomic and miRNA profiling results may provide important insights into the development of gallbladder cancer.

A Cell-Penetrant Peptide Disrupting the Transcription Factor CP2c Complexes Induces Cancer-Specific Synthetic Lethality.

Despite advances in precision oncology, cancer remains a global public health issue. In this report, proof-of-principle evidence is presented that a cell-penetrable peptide (ACP52C) dissociates transcription factor CP2c complexes and induces apoptosis in most CP2c oncogene-addicted cancer cells through transcription activity-independent mechanisms. CP2cs dissociated from complexes directly interact with and degrade YY1, leading to apoptosis via the MDM2-p53 pathway. The liberated CP2cs also inhibit TDP2, causing intrinsic genome-wide DNA strand breaks and subsequent catastrophic DNA damage responses. These two mechanisms are independent of cancer driver mutations but are hindered by high MDM2 p60 expression. However, resistance to ACP52C mediated by MDM2 p60 can be sensitized by CASP2 inhibition. Additionally, derivatives of ACP52C conjugated with fatty acid alone or with a CASP2 inhibiting peptide show improved pharmacokinetics and reduced cancer burden, even in ACP52C-resistant cancers. This study enhances the understanding of ACP52C-induced cancer-specific apoptosis induction and supports the use of ACP52C in anticancer drug development.

Widespread 8-oxoguanine modifications of miRNA seeds differentially regulate redox-dependent cancer development.

Oxidative stress contributes to tumourigenesis by altering gene expression. One accompanying modification, 8-oxoguanine (o8G) can change RNA-RNA interactions via o8G•A base pairing, but its regulatory roles remain elusive. Here, on the basis of o8G-induced guanine-to-thymine (o8G > T) variations featured in sequencing, we discovered widespread position-specific o8Gs in tumour microRNAs, preferentially oxidized towards 5' end seed regions (positions 2-8) with clustered sequence patterns and clinically associated with patients in lower-grade gliomas and liver hepatocellular carcinoma. We validated that o8G at position 4 of miR-124 (4o8G-miR-124) and 4o8G-let-7 suppress lower-grade gliomas, whereas 3o8G-miR-122 and 4o8G-let-7 promote malignancy of liver hepatocellular carcinoma by redirecting the target transcriptome to oncogenic regulatory pathways. Stepwise oxidation from tumour-promoting 3o8G-miR-122 to tumour-suppressing 2,3o8G-miR-122 occurs and its specific modulation in mouse liver effectively attenuates diethylnitrosamine-induced hepatocarcinogenesis. These findings provide resources and insights into epitranscriptional o8G regulation of microRNA functions, reprogrammed by redox changes, implicating its control for cancer treatment.

Use of Computed Tomography and Magnetic Resonance Angiograms Combined with a 3D Surgical Guide in an Elderly Cat with an Occipital Lobe Meningioma.

We present a case of occipital lobe meningioma resection in an elderly cat. The surgery was performed with the goal of avoiding major bleeding. An 11-year-old castrated indoor-only male Persian Chinchilla (5.5 kg) was presented with a month-long history of progressive tetraparesis for a left occipital lobe meningioma. Magnetic resonance imaging revealed a T2-weighted heterogeneously hyperintensity and a T1-weighted well-contrast enhancing extradural mass in the left occipital lobe of the brain. Cerebral angiographic data were obtained using magnetic resonance (MRA) and computed tomography angiography (CTA). Advanced angiograms and virtual reconstruction of images revealed that the tumor was surrounded by the caudal parasagittal meningeal vein. A left caudal rostrotentorial craniotomy and en bloc resection of the tumor were performed, and histopathology revealed a meningioma. Complete neurological recovery was achieved within 10 days after surgery. To the best of our knowledge, this is the first case report describing CTA and MRA findings and favorable clinical outcomes after surgical management of a brain meningioma without severe perioperative complications.

Human chemically-derived hepatic progenitors (hCdHs) as a source of liver organoid generation: Application in regenerative medicine, disease modeling, and toxicology testing.

BACKGROUND & AIMS: Several types of human stem cells from embryonic (ESCs) and induced pluripotent (iPSCs) to adult tissue-specific stem cells are commonly used to generate 3D liver organoids for modeling tissue physiology and disease. We have recently established a protocol for direct conversion of primary human hepatocytes (hPHs) from healthy donor livers into bipotent progenitor cells (hCdHs). Here we extended this culture system to generate hCdH-derived liver organoids for diverse biomedical applications. METHODS: To obtain hCdHs, hPHs were cultured in reprogramming medium containing A83-01 and CHIR99021 for 7 days. Liver organoids were established from hCdHs (hCdHOs) and human liver cells (hLOs) using the same donor livers for direct comparison, as well as from hiPSCs. Organoid properties were analyzed by standard in vitro assays. Molecular changes were determined by RT-qPCR and RNA-seq. Clinical relevance was evaluated by transplantation into FRG mice, modeling of alcohol-related liver disease (ARLD), and in vitro drug-toxicity tests. RESULTS: hCdHs were clonally expanded as organoid cultures with low variability between starting hCdH lines. Similar to the hLOs, hCdHOs stably maintained stem cell phenotype based on accepted criteria. However, hCdHOs had an advantage over hLOs in terms of EpCAM expression, efficiency of organoid generation and capacity for directed hepatic differentiation as judged by molecular profiling, albumin secretion, glycogen accumulation, and CYP450 activities. Accordingly, FRG mice transplanted with hCdHOs survived longer than mice injected with hLOs. When exposed to ethanol, hCdHOs developed stronger ARLD phenotype than hLOs as evidenced by transcriptional profiling, lipid accumulation and mitochondrial dysfunction. In drug-induced injury assays in vitro, hCdHOs showed a similar or higher sensitivity response than hPHs. CONCLUSION: hCdHOs provide a novel patient-specific stem cell-based platform for regenerative medicine, toxicology testing and modeling liver diseases.

Endoscopy-assisted resection of a sphenoid-wing meningioma using a 3D-printed patient-specific pointer in a dog: A case report.

A 9-year-old female mixed-breed dog presented for treatment of a presumed sphenoid-wing meningioma. Clinical signs included tonic-clonic seizures lasting <1 min, which had started 3 months previously. The physical examination results were unremarkable. An eccentrically located neoplastic cystic structure in the right sphenoid bone region suggestive of a meningioma and peritumoural brain oedema was observed in pre-operative magnetic resonance imaging (MRI). Prior to surgery, a three-dimensional (3D) patient-specific pointer (PSP) was designed using computed tomography (CT) images and computer-aided 3D design software. After a targeted approach and exposure of the lateral part of the right temporal lobe by a craniectomy guided by the 3D-PSP, complete macroscopic piecemeal resection of the meningioma could be performed using endoscopy-assisted brain surgery. Post-operative MRI confirmed complete excision of the tumor. Anticonvulsive therapy was discontinued after 90 days, and the dosage of anticonvulsants was tapered 2 weeks after surgery. At a follow-up examination 225 days post-operatively, recurrence of seizures was not observed, and the absence of tumor recurrence was confirmed by a repeat MRI examination. To the best of our knowledge, this is the first report in veterinary medicine describing a successful resection of a sphenoid-wing meningioma using a 3D-PSP. 3D-PSP-assisted craniectomy may be a surgical option for some canine skull-based tumors, such as sphenoid wing meningiomas.

Case report: Three-dimensionally printed patient-specific acetabular cage for revision surgery of aseptic loosening in a dog with micro total hip replacement.

A 2-year-old castrated male Pomeranian dog was presented for regular follow-up after micro total hip replacement (mTHR) 16 months prior to presentation. Clinically, the dog did not show any noticeable lameness of the left hindlimb, except for external rotation during walking. However, radiographic findings, namely rotation and medialization of the acetabular cup with a periprosthetic lucent line and bone formation medial to the acetabulum, were interpreted as aseptic loosening of the acetabular component. Because the dog was incompatible with the conventional THR revision method owing to severe bone defects in the acetabulum, a patient-specific titanium acetabular cage prosthesis with biflanges and four cranial and one caudal screw hole was designed for revision surgery. A custom-made acetabular cage was prepared, and it had a 12-mm polyethylene cup fixed with polymethylmethacrylate bone cement and positioned in the acetabulum. After the custom-made acetabular cage was anchored to the pelvic bone with the five cortical screws, reduction of the prostheses was achieved smoothly. The dog showed almost normal limb function without external rotation of the left hindlimb 2 weeks postoperatively. Bone remodeling and stable implant position were noted on radiographic images 3 years after revision surgery, with no evidence of loosening. Based on the clinical outcomes, the use of a custom-made acetabular prosthesis can be an effective treatment option for revision arthroplasty in acetabula with severe bone loss and structural changes in small-breed dogs.

Multiple Meningioma Resection by Bilateral Extended Rostrotentorial Craniotomy with a 3D-Print Guide in a Cat.

A 13-year-old castrated male domestic shorthair cat was referred for the surgical removal of multiple meningiomas. The cat experienced generalized tonic-clonic seizures, altered mentation, mild proprioceptive ataxia, and circling. Magnetic resonance imaging (MRI) revealed two round, solitary, well-delineated, space-occupying lesions suggestive of multiple meningiomas in the right frontal and occipital lobes. Before surgery, patient-specific three-dimensional (3D) printed models and guides were produced using a 3D program based on MRI and computed tomography (CT), and a rehearsal surgery was performed. With a 3D guide to find the location of the craniotomy lines, bilateral extended rostrotentorial craniotomy allowed en bloc resection of multiple meningiomas. The bone fragment was replaced and secured to the skull with a craniofacial plate and screws with an artificial dura. All of the surgical steps were performed without complications. The preoperative presenting signs were resolved by the time of follow-up examinations 2 weeks after surgery. Twelve months after the removal of the multiple meningiomas, the cat survived without further neurological progression. For the resection of multiple meningiomas, surgery can result in large bone defects and risk of massive hemorrhage. For this challenging surgery, patient-specific 3D models and guides can be effective for accurate and safe craniotomies.

Case report: Primary chronic calcaneal bursitis treated with subtotal bursectomy in a cat.

A 6-year-old, female spayed Bengal cat with a bodyweight of 6.4 kg was presented with swelling of the bilateral calcaneal region and weight-bearing hindlimb lameness with a 4-month history of unsuccessful conservative therapy. On orthopedic examination, a cyst-like mass around the calcaneal tendon was palpated. Palpating the mass and flexing the tarsal joint triggered pain. Through ultrasonography and magnetic resonance imaging, an inflamed or fluid-accumulated lesion was suspected around the calcaneal tendon, but there was no evidence of calcaneal tendonitis. Swollen calcaneal bursae were removed surgically. Histopathologic examination revealed fibrosis and an edematous feature. The cat was diagnosed with bilateral chronic primary calcaneal bursitis based on history, clinical signs, and diagnostic results. Hence, subtotal bursectomy was performed. At 4 weeks postoperatively, the cat had no pain around the tarsal joints and was ambulating normally. Radiographic and ultrasonographic exams revealed no recurrence of swelling or inflammation in the calcaneal region. Thirteen-month follow-up confirmed acceptable function and no relapse of clinical signs. The inflammation of calcaneal bursa alone can be the primary cause of hindlimb lameness in cats. A cat with hindlimb lameness and swelling on the calcaneal region should be assessed with the possibility of primary calcaneal bursitis. Subtotal calcaneal bursectomy can be considered as an effective treatment for primary chronic bursitis.

Epineural Neurorrhaphy of a Large Nerve Defect Due to IatroGenic Sciatic Nerve Injury in a Maltese Dog.

Epineural neurorrhaphy is a standard nerve repair method, but it is rarely reported in veterinary literature. Epineural neurorrhaphy in canine sciatic nerve injury are described in this report. An 11-month-old, castrated male Maltese dog, presented with an one-month history of non-weight bearing lameness and knuckling of the right pelvic limb. The dog showed absence of superficial and deep pain perception on the dorsal and lateral surfaces below the stifle joint. The dog had undergone femoral head and neck osteotomy in the right pelvic limb one month prior to referral at a local hospital. Based on physical and neurological examinations, peripheral nerve injury of the right pelvic limb was suspected. Radiography showed irregular bony proliferation around the excised femoral neck. Magnetic resonance imaging revealed sciatic nerve injury with inconspicuous continuity at the greater trochanter level. A sciatic nerve neurotmesis was suspected and surgical repair was decided. During surgery, non-viable tissue of the sciatic nerve was debrided, and epineural neurorrhaphy was performed to bridge a large, 20-mm defect. The superficial and deep pain perception was progressively improved and restored at 3 weeks postoperatively, and the dog exhibited a gradual improvement in motor function. At 10 weeks postoperatively, the dog showed no neurological deficit including knuckling but the tarsal joint hyperextension did not improve due to ankylosis. The dog had undergone tarsal arthrodesis and exhibited almost normal limb function without any neurologic sequela until the last follow-up at 2.5 years postoperatively.

Pelvic Endoprosthesis after Hemipelvectomy Using a 3D-Printed Osteotomy Guide for Infiltrative Osteoma in a Cat.

With the development of 3D printing and surgical techniques, various defect reconstruction methods after tumor resection have been applied not only in humans but also in veterinary medicine. This report describes a case of reconstruction after hemipelvectomy for an osteoma in a cat using a 3D-printed pelvic endoprosthesis and micro total hip replacement (mTHR). A 5-year-old spayed female Turkish Angora cat was referred for a 1-month history of constipation and intermittent weight-bearing lameness in the left hindlimb. An osteoma in the pelvis measuring 4.5 × 3 × 5.4 cm was identified based on diagnostic examinations. A left mid-to-caudal partial and right caudal partial hemipelvectomy, and a left femoral head and neck osteotomy, were planned to remove the mass. Reconstruction of the bone defect using 3D-printed metal endoprosthesis and mTHR in the left hindlimb was intended. During right caudal partial hemipelvectomy, right femoral head and neck osteotomy was performed because there was infiltration in the medial wall of the acetabulum. Histopathological examination confirmed the diagnosis of an osteoma. Two weeks post-surgery, surgical debridement and femoral stem removal were performed because of delayed wound healing and sciatic neurapraxia, leading to femoral stem dislocation from the cup. The delayed wound healing and sciatic neurapraxia were appropriately addressed. The cat regained normal weight and defecation 4 weeks post-operatively. Two years post-surgery, the patient recovered with an almost normal gait. Hemipelvectomy with 3D-printed endoprosthesis provides a safe surgical option with favorable outcomes for neoplasms in the pelvis of cats.

UPF1 Inhibits Hepatocellular Carcinoma Growth through DUSP1/p53 Signal Pathway.

Human hepatocellular carcinoma (HCC) has a high mortality rate because of the dearth of effective treatments. Multiple studies have shown that overexpression of UPF1, a key nonsense-mediated mRNA decay (NMD) factor, reduces HCC growth through various cell signaling pathways. However, the mechanism by which UPF1 expression retards HCC proliferation through the regulation of RNA stability remains unclear. By employing various UPF1 variants and transcriptome analysis, we revealed that overexpression of UPF1 variants, not UPF1-mediated NMD, reduces HCC tumorigenesis. Additionally, UPF1 variant overexpression reduced tumorigenesis in xenografted mice. Transcriptome analysis indicated that the level of dual specificity phosphatase 1 (DUSP1) was increased by UPF1 variants via posttranscriptional regulation. The UPF1 overexpression-mediated increase of DUSP1 activated tumor suppressor signaling, ultimately inhibiting cell growth. In this study, we highlighted the function of UPF1 as a tumor suppressor in HCC growth.

Transplantation of patient-specific bile duct bioengineered with chemically reprogrammed and microtopographically differentiated cells.

Cholangiopathy is a diverse spectrum of chronic progressive bile duct disorders with limited treatment options and dismal outcomes. Scaffold- and stem cell-based tissue engineering technologies hold great promise for reconstructive surgery and tissue repair. Here, we report a combined application of 3D scaffold fabrication and reprogramming of patient-specific human hepatocytes to produce implantable artificial tissues that imitate the mechanical and biological properties of native bile ducts. The human chemically derived hepatic progenitor cells (hCdHs) were generated using two small molecules A83-01 and CHIR99021 and seeded inside the tubular scaffold engineered as a synergistic combination of two layers. The inner electrospun fibrous layer was made of nanoscale-macroscale polycaprolactone fibers acting to promote the hCdHs attachment and differentiation, while the outer microporous foam layer served to increase mechanical stability. The two layers of fiber and foam were fused robustly together thus creating coordinated mechanical flexibility to exclude any possible breaking during surgery. The gene expression profiling and histochemical assessment confirmed that hCdHs acquired the biliary epithelial phenotype and filled the entire surface of the fibrous matrix after 2 weeks of growth in the cholangiocyte differentiation medium in vitro. The fabricated construct replaced the macroscopic part of the common bile duct (CBD) and re-stored the bile flow in a rabbit model of acute CBD injury. Animals that received the acellular scaffolds did not survive after the replacement surgery. Thus, the artificial bile duct constructs populated with patient-specific hepatic progenitor cells could provide a scalable and compatible platform for treating bile duct diseases.

Generation and differentiation of chemically derived hepatic progenitors from mouse primary hepatocytes.

This protocol describes the generation of bipotent chemically derived hepatic progenitors (mCdHs) from mouse primary hepatocytes and their subsequent differentiation into either hepatic or cholangiocytic lineages. The reprogrammed mCdHs have a high proliferation capacity and express progenitor markers in long-term passages. Differentiated mCdHs show the characteristics of either hepatic or cholangiocytic genes. This protocol has potential application for regenerative medicine, including ex vivo gene therapy, disease modeling, drug screening, and personalized medicine. For complete details on the use and execution of this protocol, please refer to Kim et al. (2021).

Association of Microbial Dysbiosis with Gallbladder Diseases Identified by Bile Microbiome Profiling.

BACKGROUND: Cholecystitis is an important risk factor for gallbladder cancer, but the bile microbiome and its association with gallbladder disease has not been investigated fully. We aimed to analyze the bile microbiome in normal conditions, chronic cholecystitis, and gallbladder cancer, and to identify candidate bacteria that play an important role in gallbladder carcinogenesis. METHODS: We performed metagenome sequencing on bile samples of 10 healthy individuals, 10 patients with chronic cholecystitis, and 5 patients with gallbladder cancer, and compared the clinical, radiological, and pathological characteristics of the participants. RESULTS: No significant bacterial signal was identified in the normal bile. The predominant dysbiotic bacteria in both chronic cholecystitis and gallbladder cancer were those belonging to the Enterobacteriaceae family. Klebsiella increased significantly in the order of normal, chronic cholecystitis, and gallbladder cancer. Patients with chronic cholecystitis and dysbiotic microbiome patterns had larger gallstones and showed marked epithelial atypia, which are considered as precancerous conditions. CONCLUSION: We investigated the bile microbiome in normal, chronic cholecystitis, and gallbladder cancer. We suggest possible roles of Enterobacteriaceae, including Klebsiella, in gallbladder carcinogenesis. Our findings reveal a possible link between a dysbiotic bile microbiome and the development of chronic calculous cholecystitis and gallbladder cancer.