Inflammatory myofibroblastic tumor in the urinary bladder in a dog.

An 8-year-old castrated male Maltese dog was presented with a urinary bladder mass, urolithiasis, and hematuria. A solitary, pedunculated, intraluminal mass on the caudodorsal wall was identified with extensive irregular bladder wall thickening, and the mass was surgically removed. Postoperative histopathology demonstrated a submucosal lesion comprising spindle cells with marked inflammatory cell infiltration, without malignant changes. Immunohistochemical staining revealed vimentin and desmin positivity in the mass. An inflammatory myofibroblastic tumor (IMT) was definitively diagnosed. No recurrence was observed during a 43-month follow-up period. Although IMTs are rare in dogs, they should be considered a differential diagnosis for mass-like urinary bladder lesions accompanying a chronic inflammatory disease process. Key clinical message: Canine IMT should be included in the differential diagnoses of bladder masses, especially when dogs exhibit chronic irritation and inflammation.

Tumeur myofibroblastique inflammatoire de la vessie chez un chienUn chien maltais mâle castré de 8 ans a été présenté avec une masse à la vessie, une lithiase urinaire et une hématurie. Une masse intraluminale pédonculée solitaire sur la paroi caudodorsale a été identifiée avec un épaississement important et irrégulier de la paroi vésicale, et la masse a été retirée chirurgicalement. L'histopathologie postopératoire a mis en évidence une lésion à la sous-muqueuse comprenant des cellules fusiformes avec une infiltration cellulaire inflammatoire marquée, sans modification maligne. La coloration immunohistochimique a révélé une positivité à la vimentine et à la desmine dans la masse. Une tumeur myofibroblastique inflammatoire (IMT) a été définitivement diagnostiquée. Aucune récidive n'a été observée au cours d'une période de suivi de 43 mois. Bien que les IMT soient rares chez le chien, ils doivent être considérés comme un diagnostic différentiel des lésions de la vessie de type masse accompagnant un processus de maladie inflammatoire chronique.Message clinique clé:L'IMT canine doit être incluse dans les diagnostics différentiels des masses vésicales, en particulier lorsque les chiens présentent une irritation et une inflammation chroniques.(Traduit par Dr Serge Messier).

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.

Thyroid carcinosarcoma masquerading as a paraesophageal abscess in a Cane Corso dog.

A 12-year-old castrated male Cane Corso dog was presented with cervical swelling, lethargy, anorexia, and cough. An extensive neck mass with necrotic cysts was observed, severely adherent to surrounding tissues. Based on diagnostic imaging including ultrasound, computed tomography, and fine-needle aspiration cytology, paraesophageal abscess was tentatively diagnosed. However, after the mass was surgically removed, a diagnosis of thyroid carcinosarcoma composed of neoplastic cell populations with epithelial and mesenchymal origins was made via histopathology and immunohistochemistry. The dog died of a recurrent mass with pulmonary metastasis 105 d after surgery. This report describes a rare type of canine thyroid cancer, thyroid carcinosarcoma, preoperatively masquerading as an abscess and postoperatively confirmed by histopathology. Key clinical message: Thyroid carcinosarcoma, despite its rarity in dogs, should be included in the differential diagnoses of cervical mass especially with an aggressive progression.

Carcinosarcome thyroïdien déguisé en abcès paraoesophagien chez un chien Cane Corso. Un chien Cane Corso mâle castré de 12 ans a été présenté avec de l'enflure cervicale, de la léthargie, de l'anorexie et une toux. Une masse étendue du cou avec des kystes nécrotiques a été observée, adhérente fortement aux tissus environnants. Sur la base de l'imagerie diagnostique comprenant l'échographie, la tomodensitométrie et la cytologie par aspiration à l'aiguille fine, un abcès paraoesophagien a été provisoirement diagnostiqué. Cependant, après l'ablation chirurgicale de la masse, un diagnostic de carcinosarcome thyroïdien composé de populations de cellules néoplasiques d'origine épithéliale et mésenchymateuse a été posé par histopathologie et immunohistochimie. Le chien est décédé d'une masse récurrente avec métastase pulmonaire 105 jours après la chirurgie. Ce rapport décrit un type rare de cancer de la thyroïde canine, le carcinosarcome thyroïdien, se faisant passer pour un abcès en préopératoire et confirmé en postopératoire par histopathologie.Message clinique clé:Le carcinosarcome thyroïdien, malgré sa rareté chez le chien, doit être inclus dans les diagnostics différentiels de masse cervicale surtout à évolution agressive.(Traduit par Dr Serge Messier).

Disseminated fungal infection with Aspergillus versicolor and Schizophyllum commune in a dog.

A two-year-old neutered male Coton de Tulear presented with lethargy, anorexia, and tachypnea. Cystic masses noticed at the cranial mediastinal region were diagnosed as granuloma containing hyphae of Aspergillus versicolor. Despite antifungal treatment using itraconazole, fluconazole, and voriconazole, the lesions spread to the lung. After euthanasia, Schizophyllum commune was identified in the lung and splenic lymph node. This is the first case of fungal infection caused by A. versicolor and S. commune in a dog.

Expression of the hedgehog signalling pathway and the effect of inhibition at the level of smoothened in canine osteosarcoma cell lines.

Osteosarcoma (OSA) is the most common malignant bone cancer in dogs. Canine and human OSA share several features, including tumour environments, response to traditional treatment, and several molecular pathways. Hedgehog (Hh) signalling is known to contribute to tumorigenesis and progression of various cancers, including human OSA. This study aimed to identify the role of the Hh signalling pathway in canine OSA cell lines, including Abrams, D17, and Moresco, focusing on the signal transducer Smoothened (SMO). mRNA and protein levels of Hh pathway components, including SHH, IHH, SMO, and PTCH1, were aberrant in all examined OSA cell lines compared with canine osteoblast cells. The SMO inhibitor cyclopamine significantly decreased cell viability and colony-forming ability in the canine OSA cell lines in a dose-dependent manner. Moresco cells, which expressed the highest level of SMO protein, were the most sensitive to the anticancer effect of cyclopamine among the three canine OSA cell lines tested. Hh downstream target gene and protein expression in canine OSA cell lines were downregulated after cyclopamine treatment. In addition, cyclopamine significantly increased apoptotic cell death in Abrams and Moresco cells. The findings that Hh/SMO is activated in canine OSA cell lines and cyclopamine suppresses OSA cell survival via inhibition of SMO suggest that the Hh/SMO signalling pathway might be a novel therapeutic target for canine OSA.

Leiomyosarcoma of urinary bladder in a Shih Tzu dog.

A 10-year-old intact male Shih Tzu dog presented with hematuria. Double-contrast cystography revealed a polypoid filling defect at the apex of the urinary bladder. Ultrasonography revealed a heterogeneously hypoechoic intramural mass with minimal vascular flow beneath the submucosal layer. After partial cystectomy, a well-demarcated bladder leiomyosarcoma was diagnosed on histopathology. The patient was alive and well without any clinical signs, recurrence, or metastasis at the 29-month follow-up after the surgical removal of the bladder mass. Leiomyosarcoma should be considered as a differential diagnosis if mass-like lesions are observed in the urinary bladder, although this type of malignancy is rare in canines. Histopathological confirmation is important for predicting prognosis and determining further medical plans.

Surgical management of a metastatic vertebral tumour originating from a mammary adenocarcinoma in a dog.

An 11-year-old spayed female Toy Poodle presented with acute tetraparesis. A small subcutaneous mass was found in the right trunk region, and the magnetic resonance revealed a compressive spinal cord lesion due to an irregular bone proliferation at the third cervical vertebra. After surgical resection of the vertebral lesion, the neurological symptoms improved, and the patient could walk on her own. The excised vertebral and subcutaneous masses were diagnosed as a mammary adenocarcinoma on the histopathological examination, with Ki-67 and HER-2 immunohistochemistry staining. This case report highlights the importance of defining the primary tumours of metastatic vertebral tumours and the necessity of palliative surgery to improve the patient's quality of life.

Effect of 7.5% hypertonic saline solution on whole blood coagulation in healthy dogs using thromboelastography.

OBJECTIVE: To evaluate the effects of 7.5% hypertonic saline solution (HSS) on whole blood coagulation in healthy dogs and to compare electrolyte and osmolality measurements between in vivo and in vitro dilution with HSS. DESIGN: Experimental study. SETTING: University teaching hospital. ANIMALS: Twelve adult purpose-bred Beagles. INTERVENTIONS: All 12 dogs received 5 mL/kg 7.5% HSS at 1 mL/kg/min. After a 14-day washout period, 5 of these dogs were randomly selected and received the same volume of 0.9% NaCl. Blood samples were collected before infusion, immediately after infusion, and at 30, 60, and 90 minutes after infusion for the measurement of coagulation using thromboelastography. For comparison of electrolyte concentrations and osmolality between in vitro dilution and in vivo dilution of HSS, 6-mL blood samples were diluted with 7.5% HSS (1:18 ratio) at baseline. MEASUREMENTS AND MAIN RESULTS: None of the thromboelastography variables differed significantly between the 7.5% HSS group and the 0.9% NaCl group. The sodium and chloride levels, and the osmolality, were significantly increased at all postinfusion time points compared to baseline, while those levels were significantly higher with in vitro dilution than all postinfusion time points. However, almost all the values gradually decreased and became similar to baseline values in case of in vivo dilution. CONCLUSIONS: The clinically relevant dose of 7.5% HSS (5 mL/kg) did not affect whole blood coagulation significantly in healthy Beagles. Further studies are necessary to assess the effect of HSS on blood coagulation in canine patients with shock.

TSG-6 in extracellular vesicles from canine mesenchymal stem/stromal is a major factor in relieving DSS-induced colitis.

Adipose tissue derived mesenchymal stem/stromal cell (ASC)-derived extracellular vesicles (EV) have been reported to be beneficial against dextran sulfate sodium (DSS)-induced colitis in mice. However, the underlying mechanisms have not been fully elucidated. We hypothesize that the tumor necrosis factor-α-stimulated gene/protein 6 (TSG-6) in EVs is a key factor influencing the alleviation of colitis symptoms. DSS-induced colitis mice (C57BL/6, male, Naïve = 6, Sham = 8, PBS = 8 EV = 8, CTL-EV = 8, TSG-6 depleted EV = 8) were intraperitoneally administered EVs (100 ug/mice) on day 1, 3, and 5; colon tissues were collected on day 10 for histopathological, RT-qPCR, western blot and immunofluorescence analyses. In mice injected with EV, inflammation was alleviated. Indeed, EVs regulated the levels of pro- and anti-inflammatory cytokines, such as TNF-α, IL-1ß, IFN-γ, IL-6, and IL-10 in inflamed colons. However, when injected with TSG-6 depleted EV, the degree of inflammatory relief was reduced. Furthermore, TSG-6 in EVs plays a key role in increasing regulatory T cells (Tregs) and polarizing macrophage from M1 to M2 in the colon. In conclusion, this study shows that TSG-6 in EVs is a major factor in the relief of DSS-induced colitis, by increasing the number of Tregs and macrophage polarization from M1 to M2 in the colon.

Comparison of native and citrated whole blood samples for rapid thromboelastography in Beagles.

OBJECTIVE: To examine the extent to which rapid thromboelastography (r-TEG) could decrease the testing time in comparison with that required for kaolin-activated thromboelastography (TEG), and to compare 2 types of blood samples (ie, native and citrated whole blood [WB]), for determining r-TEG values in healthy dogs. DESIGN: Prospective observational study. SETTING: University teaching hospital. ANIMALS: Sixteen healthy Beagles. INTERVENTIONS: Kaolin-activated TEG test using citrated WB samples and r-TEG test using native and citrated WB samples were performed in 16 dogs. At 60 minutes after the initial blood sampling, further samples were collected from a subset of 6 dogs in the same manner to evaluate intraindividual repeatability of r-TEG. MEASUREMENTS AND MAIN RESULTS: The mean time to maximum amplitude (MA) for r-TEG with native and citrated WB samples was recorded as 1313.9 ± 250.9 seconds and 1351.3 ± 264.6 seconds (mean ± SD), respectively, and 1779.9 ± 197.0 seconds for kaolin-activated TEG. Coefficients of variation with native and citrated WB samples for r-TEG values, TEG-activated clotting time, clot formation time, α angle, and MA, were determined to be 13.4% versus 18.8%, 11.1% versus 16.6%, 4.2% versus 5.1%, and 10.0% versus 10.0%, respectively. Intraindividual variations were lower for native WB samples than for citrated WB samples. CONCLUSIONS: The r-TEG test significantly decreased the mean time to MA compared with the kaolin-activated TEG test. In addition, native WB samples showed lower coefficients of variation and intraindividual variation than citrated WB samples in r-TEG analysis; this suggests that native WB samples can provide more consistent results. Therefore, the r-TEG method using native WB samples is recommended for assessment of dogs' hemostatic status when an early diagnosis is required.

Preconditioning of canine adipose tissue-derived mesenchymal stem cells with deferoxamine potentiates anti-inflammatory effects by directing/reprogramming M2 macrophage polarization.

Preconditioning with hypoxia or hypoxia-mimetic agents has been tried with mesenchymal stem cells (MSCs) to improve the secretion of anti-inflammatory factors. These preconditioning procedures upregulate hypoxia inducible factor (HIF) 1-alpha leading to the transcription of HIF-dependent tissue protective and anti-inflammatory genes. Due to the limited number of studies exploring the activity of deferoxamine (DFO)-a hypoxia-mimetic agent-in MSCs, we aimed to determine whether DFO can enhance the secretion of anti-inflammatory substances in canine adipose tissue-derived (cAT)-MSCs. Furthermore, we investigated whether this activity of DFO could affect macrophage polarization and activate anti-inflammatory reactions. cAT-MSCs preconditioned with DFO exhibited enhanced secretion of anti-inflammatory factors such as prostaglandin E2 and tumor necrosis factor-α-stimulated gene-6. To evaluate the interaction between DFO preconditioned cAT-MSCs and macrophages, RAW 264.7 cells were co-cultured with cAT-MSCs using the Transwell system, and changes in the expression of factors related to macrophage polarization were analyzed using the quantitative real-time PCR and western blot assays. When RAW 264.7 cells were co-cultured with DFO preconditioned cAT-MSCs, the expression of M1 and M2 markers decreased and increased, respectively, compared to co-culturing with non-preconditioned cAT-MSCs. Thus, cAT-MSCs preconditioned with DFO can more effectively direct and reprogram macrophage polarization into the M2 phase, an anti-inflammatory state.

Antitumour effects of Liporaxel (oral paclitaxel) for canine melanoma in a mouse xenograft model.

Paclitaxel, a member of the taxane family, exhibits antitumour effects by targeting the microtubules in cancer cells. Recently, oral paclitaxel has been developed to overcome the side effects of intravenous paclitaxel administration in human patients. The objective of this study was to investigate the antitumour effects of oral paclitaxel in vitro and in vivo. Three weeks after inoculation, oral paclitaxel (25 and 50 mg/kg) or saline was administered every week for three consecutive weeks. To explore the underlying mechanism, tumour angiogenesis was examined by immunohistochemistry with an anti-CD31 antibody. Tumour cell apoptosis was detected by Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling assay, and cell cycle arrest was confirmed by western blot analysis. Oral paclitaxel treatment of canine melanoma cells exerted mediated antiproliferative effects and mediated cell cycle arrest in vitro. In animal experiments, after oral paclitaxel administration, the average tumour size decreased to approximately 30% of that in the control. Histologically, oral paclitaxel showed anti-angiogenic effects and induced the apoptosis in tumour tissues. Oral paclitaxel also downregulated the intratumoural expression of cyclin D1 and inhibited cell proliferation. The study findings support potential application of oral paclitaxel as a novel chemotherapeutic strategy to treat canine melanoma. This is the first study to investigate the potential of oral paclitaxel as a therapeutic drug against canine tumours.

Melarsomine suppresses canine osteosarcoma cell survival via inhibition of Hedgehog-GLI signaling.

The Hedgehog-GLI signaling pathway is activated in human and canine osteosarcoma (OSA) and represents a potential therapeutic target for cancers, including OSA. Arsenic trioxide represses GLI expression. Melarsomine, an arsenic compound-containing drug, has been approved for the treatment of canine heartworm disease. Hence, we hypothesized that melarsomine inhibits GLI signaling in canine OSA cell lines. The present study aimed to assess this hypothesis. Cell viability and colony formation were decreased in the canine OSA cell lines Abrams and D17 after treatment with melarsomine. Melarsomine-induced apoptotic cell death was assessed via cell cycle analysis using propidium iodide staining. Quantitative real-time reverse transcription polymerase chain reaction and western blot analyses revealed a downregulation of genes downstream of the Hedgehog signaling pathway, including GLI1, GLI2, and PTCH, after melarsomine treatment. The present results suggest that melarsomine exerts antitumor effects and serves as a GLI inhibitor in canine OSA cells. Additional studies are required to evaluate and confirm the anticancer effect and relevant therapeutic dose of melarsomine in vivo.

Enhanced angiogenic activity of dimethyloxalylglycine-treated canine adipose tissue-derived mesenchymal stem cells.

The paracrine function of mesenchymal stem cells (MSCs) during transplantation has been recently studied due to its poor differentiation ratio. Dimethyloxalylglycine (DMOG) has been used to promote angiogenesis in experimental animal models, however, comparable approaches for canine MSCs are not sufficient. In the present study, we assessed whether DMOG improves angiogenesis in canine adipose tissue-derived mesenchymal stem cells (cAT-MSCs). cAT-MSCs were treated with DMOG and their effect on angiogenesis was investigated by cell proliferation assay, western blotting, and tube formation assay. Dimethyloxalylglycine preconditioning enhanced the expression of vascular endothelial growth factor (VEGF) among pro-angiogenic factors in cAT-MSCs via hypoxia-inducible factor-1α stabilization. Dimethyloxalylglycine primed-cAT-MSC-conditioned media increased angiogenesis in human umbilical vein endothelial cells. These results suggest that DMOG conditioning of cAT-MSCs augmented the secretion of VEGF, which acted as a prominent pro-angiogenic factor during angiogenesis. DMOG-primed cAT-MSCs may have the potential to induce beneficial effects in ischemic diseases in clinical trials.

Canine adipose tissue-derived mesenchymal stem cells pre-treated with TNF-alpha enhance immunomodulatory effects in inflammatory bowel disease in mice.

Canine inflammatory bowel disease (IBD) is an intractable autoimmune disorder that results in various gastrointestinal and systemic symptoms. Mesenchymal stem cells (MSCs), which release immunomodulatory factors such as tumor necrosis factor-α (TNF-α)-induced gene/protein 6 (TSG-6) and prostaglandin E2 (PGE2), have been suggested as an alternative therapeutic option for IBD treatment in veterinary medicine. Furthermore, although it is known that MSCs pre-treated with pro-inflammatory cytokines show enhanced anti-inflammatory properties via the secretion of soluble factors, the underlying mechanisms of IBD remain unclear. The aim of this study was to demonstrate the therapeutic effects and corresponding mechanisms of canine adipose tissue-derived (cAT)-MSCs stimulated with TNF-α in mouse models of IBD. Mice with dextran sulfate sodium (DSS)- or dinitrobenzene sulfonic acid (DNBS)-induced colitis were injected intraperitoneally with cAT-MSCs pre-treated with TNF-α. Colitis severity was assessed and colon tissues were collected for histopathological, enzyme-linked immunosorbent assay, and flow cytometry analysis. cAT-MSCs stimulated with TNF-α secreted higher concentrations of immunomodulatory factors such as TSG-6 and PGE2, which play a key role in inducing phenotypic alterations in macrophages. Consequently, TNF-α-pre-treated cAT-MSCs further regulated colonic inflammatory cytokines such as interleukin (IL)-1ß, IL-6, and IL-10, and ameliorated DSS- or DNBS-induced colitis in mice. Additionally, we demonstrated that M1 macrophages (F4/80+/iNOS+ cells) were decreased in colon tissues from mice treated with TNF-α-pre-treated cAT-MSCs, whereas M2 macrophages (F4/80+/CD206+ cells) were increased. These results may suggest a new cell-based therapeutic option for treating IBD.

Influence of needle gauge and venepuncture difficulty on thromboelastography in healthy cats.

OBJECTIVES: The aim of this study was to evaluate the effect of two differently sized butterfly catheter needles and the effect of venepuncture difficulty on thromboelastography (TEG) results in healthy cats. METHODS: Twenty-four healthy cats were included. Blood samples were collected from the jugular vein by syringe aspiration via direct venepuncture with 21 G and 22 G butterfly needles. The venepuncture difficulty score was classified into four categories. The first 1.5 ml blood drawn from each subject was discarded before collecting a sample for TEG analysis. TEG analyses were performed on citrated whole blood samples from 17 clinically healthy cats, using assays with kaolin as activators. Among the TEG parameters, reaction time (R), clot formation time (κ), alpha angle (α), maximum amplitude (MA) and global clot strength (G) were recorded from each tracing. RESULTS: Seven cats were excluded from the study; results were obtained for the remaining 17 cats. There were no statistically significant differences between the use of two different needles for R (P = 0.72), κ (P = 0.74), α (P = 0.99), MA (P = 0.08) and G (P = 0.09). Samples with difficulty scores ⩾1 were not significantly different from samples with difficulty scores of 0 for R (P = 0.24), κ (P = 0.65), α (P = 0.65), MA (P = 0.72) and G (P = 0.77). CONCLUSIONS AND RELEVANCE: The results of TEG in clinically healthy cats do not differ significantly when using two different gauge needles. There was no significant difference in the TEG results according to venepuncture difficulty scoring.

Anticancer effects of high-dose ascorbate on canine melanoma cell lines.

The use of the well-known powerful antioxidant ascorbate has recently become more widespread in human medicine. Intravenous administration of high-dose ascorbate has been demonstrated to exert anticancer effects. It has resulted in effective cell death in vitro and inhibition of tumour growth in vivo. The aim of the current study was to evaluate the effects of high-dose ascorbate on canine melanoma in vitro. Four canine melanoma cell lines, UCDK9M1, UCDK9M3, UCDK9M4 and UCDK9M5 were treated with ascorbate for 2 hours at a range of millimolar concentrations (0-20 mM) to investigate the resulting effects on cell viability. All four canine melanoma cell lines exhibited reduced viability in a dose-dependent manner. Further investigation demonstrated that high-dose ascorbate induced apoptosis via the activation of Bax. These findings suggest that high-dose ascorbate has an anticancer effect on canine melanoma cell lines in vitro. With regard to clinical application, further in vivo investigation should be conducted.

TSG-6 secreted by human adipose tissue-derived mesenchymal stem cells ameliorates severe acute pancreatitis via ER stress downregulation in mice.

BACKGROUND: Through recent studies, the onset of acute pancreatitis in pancreatic acinar cells (PACs) and the regulatory role of PACs in severe acute pancreatitis (SAP) have been revealed. During the early stages of pancreatitis, the endoplasmic reticulum (ER) in PACs undergoes significant changes, including swelling and vacuolization. In response to an increase in the extracellular stress in ER, PACs lose their functions, leading to cell apoptosis and inflammation response. The beneficial effects of human adipose tissue-derived mesenchymal stem cells (hAT-MSCs) on SAP have been well documented in previous studies. However, the underlying mechanism of their action remains controversial. METHODS: In this study, the therapeutic effects of intraperitoneally administered hAT-MSCs in a caerulein (50 µg/kg)- and lipopolysaccharide (LPS) (10 mg/kg)-co-induced SAP mouse model were evaluated. Inflammatory response and ER stress were measured in pancreatic tissue samples, and the beneficial effects were evaluated through quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blot, and immunofluorescence analysis. RESULTS: Inflammatory response and ER stress were ameliorated following hAT-MSC injection, and the beneficial effects were observed in the absence of significant engraftment of hAT-MSCs. hAT-MSCs transfected with siRNA-targeting tumour necrosis factor-α-induced gene/protein 6 (TSG-6) were unable to inhibit ER stress and inflammation. In addition, TSG-6 from hAT-MSCs significantly suppressed ER stress-induced apoptosis and nuclear factor kappa B (NF-κB) activity in SAP model mice. CONCLUSIONS: TSG-6 secreted by hAT-MSCs protects PACs in SAP model mice via the inhibition of ER stress, as well as inflammatory responses. This study has revealed a new area for ER stress-targeted therapy in SAP patients.

Evaluation of nafamostat mesilate as an alternative anticoagulant during intermittent hemodialysis in healthy Beagle dogs.

OBJECTIVE: To evaluate nafamostat mesilate (NM) as an alternative anticoagulant agent for intermittent hemodialysis (IHD). DESIGN: Prospective randomized study. SETTING: University teaching hospital. ANIMALS: Eighteen healthy Beagle dogs. INTERVENTIONS: In group 1 (n = 6), NM was administered at a dose of 0.5 mg/kg/h during IHD for 5 hours. In group 2 (n = 6), NM was administered at a low dose of 0.25 mg/kg/h during IHD. In group 3 (n = 6), which was the control group, unfractionated heparin (UFH) was administered during IHD. The evaluated parameters included: the amount of residual blood clots in the blood chamber and arterial side of the dialyzer; the levels of hemoglobin, hematocrit, and platelets; and the prothrombin time (PT), activated partial thromboplastin time (aPTT), and activated clotting time (ACT). MEASUREMENTS AND MAIN RESULTS: Groups 1 and 2 successfully completed IHD without serious coagulation in the extracorporeal circulation. The residual blood clotting in the blood chamber and arterial side of the dialyzer did not significantly differ in groups 1 and 2 compared to group 3 (group 1 vs group 3, P = 1.000; and group 2 vs group 3, P = 1.000). No significant differences were observed between pre- and posttreatment PTs in groups 1 (P = 0.476) and 2 (P = 0.597), between pre- and posttreatment aPTTs in groups 1 (P = 0.983) and 2 (P = 0.977), and between pre- and posttreatment ACT in groups 1 (P = 0.282) and 2 (P = 0.401). In group 3, a significant elevation of ACT was observed at the posttest (P < 0.001). CONCLUSIONS: The results of this study in healthy Beagle dogs suggest that NM at 0.25 mg/kg/h may be a valid alternative to UFH for IHD. Further studies are needed in patients at high risk of bleeding.