Akinori Mizoguchi

Lipoprotein lipase (LPL) plays a central role in incorporating plasma lipids into tissues and regulates lipid metabolism and energy balance in the human body. Conversely, LPL expression is almost absent in normal adult livers. Therefore, its physiological role in the liver remains unknown. We aimed to elucidate the role of LPL in the pathophysiology of nonalcoholic steatohepatitis (NASH), a hepatic manifestation of obesity. Hepatic stellate cell (HSC)-specific LPL-knockout (LplHSC-KO ) mice, LPL-floxed (Lplfl/fl ) mice, or double-mutant toll-like receptor 4-deficient (Tlr4-/- ) LplHSC-KO mice were fed a high-fat/high-cholesterol diet for 4 weeks to establish the nonalcoholic fatty liver model or an high-fat/high-cholesterol diet for 24 weeks to establish the NASH model. Human samples, derived from patients with nonalcoholic fatty liver disease, were also examined. In human and mouse NASH livers, serum obesity-related factors, such as free fatty acid, leptin, and interleukin-6, dramatically increased the expression of LPL, specifically in HSCs through signal transducer and activator of transcription 3 signaling, as opposed to that in hepatocytes or hepatic macrophages. In the NASH mouse model, liver fibrosis was significantly reduced in LplHSC-KO mice compared with that in Lplfl/fl mice. Nonenzymatic LPL-mediated cholesterol uptake from serum lipoproteins enhanced the accumulation of free cholesterol in HSCs, which amplified TLR4 signaling, resulting in the activation of HSCs and progression of hepatic fibrosis in NASH. Conclusion: The present study reveals the pathophysiological role of LPL in the liver, and furthermore, clarifies the pathophysiology in which obesity, as a background factor, exacerbates NASH. The LPL-mediated HSC activation pathway could be a promising therapeutic target for treating liver fibrosis in NASH.

Abstract Background and Aim Dietary emulsifiers are widely used in processed foods and officially approved as safe for intake. However, recent studies have demonstrated that some emulsifiers alter the colonic microbiota, leading to colonic low‐grade inflammation, in mice. The effect of dietary emulsifiers on small‐intestinal microbiota, which is important for gut immunity, has not been studied. We aimed to investigate the effect of a representative dietary emulsifier, polysorbate‐80 (P80), on the small‐intestinal microbiota in normal mice. Methods Some mice were pretreated with P80 for 8 weeks with or without indomethacin administration on the last 2 days, and intestinal damage was evaluated histologically. The ileal and colonic microbiota composition was assessed using 16S rRNA polymerase chain reaction. Results Polysorbate‐80 increased the Gammaproteobacteria abundance and decreased the α‐diversity in the small intestine. No decrease in α‐diversity was observed in the colon. P80 pretreatment exacerbated the indomethacin‐induced small‐intestinal lesions and significantly increased the interleukin‐1β expression. Culture of ileal content on deoxycholate hydrogen sulfide lactose agar showed that P80 significantly increased the colonies of the sulfide‐producing bacteria Proteus spp. (genetically identified as Proteus mirabilis ). Antibiotic pretreatment abolished the P80‐induced aggravation of indomethacin‐induced ileitis. Motility assay in semisolid agar showed that adding 0.02% P80 to the agar significantly increased the diameter of P. mirabilis colonies but not that of Escherichia coli colonies. Conclusions Polysorbate‐80 enhances the vulnerability of the small intestine to indomethacin‐induced injury by inducing ileal dysbiosis. Direct enhancement of the motility of specific flagellated microbiota by P80 might be related to dysbiosis and intestinal injury.

Aim Chitinase 3‐like 1 (CHI3L1), an 18‐glycosyl hydrolase‐related molecule, is a member of the enzymatically inactive chitinase‐like protein family. Serum levels of CHI3L1 are strongly correlated with hepatic fibrosis progression during many liver diseases. Therefore, this protein could be involved in the development of hepatic fibrosis pathology; however, its role has not been elucidated. We aimed to elucidate its role in the pathophysiology of liver fibrosis. Methods Chitinase 3‐like 1‐deficient ( Chi3l1 −/− ) mice were given carbon tetrachloride twice per week for 4 weeks or fed a methionine choline‐deficient diet for 12 weeks to generate mouse liver fibrosis models. Human fibrotic liver tissues were also examined immunohistochemically. Results In human and mouse fibrotic livers, CHI3L1 expression was mainly localized to hepatic macrophages, and the intrahepatic accumulation of CHI3L1 + macrophages was significantly enhanced compared to that in control livers. In the two mouse models, hepatic fibrosis was significantly ameliorated in Chi3l1 −/− mice compared to that in wild‐type mice, which was dependent on hepatic macrophages. The accumulation and activation of hepatic macrophages was also significantly suppressed in Chi3l1 −/− mice compared to that in wild‐type mice. Furthermore, apoptotic hepatic macrophages were significantly increased in Chi3l1 −/− mice. Chitinase 3‐like 1 was found to inhibit hepatic macrophage apoptosis by suppressing Fas expression and activating Akt signaling in an autocrine manner, which resulted in hepatic macrophage accumulation and activation, exaggerating liver fibrosis. Conclusions Chitinase 3‐like 1 exacerbates liver fibrosis progression by suppressing apoptosis in hepatic macrophages. Therefore, this might be a potential therapeutic target for the treatment of liver fibrosis.

Recent studies indicate that probiotics, which have attracted attention as a treatment for irritable bowel syndrome, affect intestinal homeostasis. In this study, we investigated whether Zygosaccharomyces sapae (strain I-6), a probiotic yeast isolated from miso (a traditional Japanese fermented food), could improve irritable bowel syndrome symptoms.Male Wistar rats were exposed to water avoidance stress (WAS). The number of defecations during WAS and the visceral hypersensitivity before and after WAS were evaluated using colorectal distension. Tight junction changes were assessed by Western blotting. Some rats were fed with strain I-6 or β-glucan from strain I-6. Changes in the intestinal microbiota were analyzed. The effect of fecal microbiota transplantation after WAS was evaluated similarly. Caco-2 cells were stimulated with interleukin-1β and tight junction changes were investigated after coculture with strain I-6.The increased number of stool pellets and visceral hypersensitivity induced by WAS were suppressed by administering strain I-6. The decrease in tight junction protein occludin by WAS was reversed by the administration of strain I-6. β-Glucan from strain I-6 also suppressed those changes induced by WAS. In the rat intestinal microbiota, treatment with strain I-6 altered the β-diversity and induced changes in bacterial occupancy. Upon fecal microbiota transplantation, some symptoms caused by WAS were ameliorated.These results suggest that traditional fermented foods such as miso in Japan are valuable sources of probiotic yeast candidates, which may be useful for preventing and treating stress-induced visceral hypersensitivity.

The artificial sweetener acesulfame potassium (ACK) is officially approved as safe for intake and has been used in processed foods. However, ACKs have been reported to induce metabolic syndrome, along with alteration of the gut microbiota in mice. In recent years, studies have suggested that this artificial sweetener promotes myeloperoxidase reactivity in Crohn's disease-like ileitis. We aimed to investigate the effect of ACK on the intestinal mucosa and gut microbiota of normal mice.Acesulfame potassium was administered to C57BL/6J mice (8 weeks old) via free drinking. Intestinal damage was evaluated histologically, and messenger RNA (mRNA) levels of TNF-α, IFN-γ, IL1-β, MAdCAM-1, GLP1R, and GLP2R were determined with quantitative reverse transcription polymerase chain reaction (qRT-PCR). Immunohistochemistry was performed to examine the expression of MAdCAM-1 in the small intestine. The composition of gut microbiota was assessed using high-throughput sequencing. We performed intravital microscopic observation to examine if ACK altered lymphocyte migration to the intestinal microvessels.Acesulfame potassium increased the expression of proinflammatory cytokines, decreased the expression of GLP-1R and GLP-2R, and induced small intestinal injury with an increase in intestinal permeability, and ACK treatment induced microbial changes, but the transfer of feces alone from ACK mice did not reproduce intestinal damage in recipient mice. ACK treatment significantly increased the migration of lymphocytes to intestinal microvessels.Acesulfame potassium induces dysbiosis and intestinal injury with enhanced lymphocyte migration to intestinal mucosa. Massive use of non-caloric artificial sweeteners may not be as safe as we think.

Although the involvement of intestinal microbiota in innate immunity has been reported recently, the pathogenicity of autoimmune pancreatitis (AIP) remains unclear. This study aimed to investigate whether probiotics ameliorate inflammation in AIP through interactions with innate immunity.

Takayasu arteritis (TA) sometimes presents with colitis, which may be diagnosed as inflammatory bowel disease unclassified (IBDU) because of atypical or mixed findings of ulcerative colitis (UC) and Crohn's disease. We herein report an 18-year-old girl presenting with colitis with an occasional high fever eventually diagnosed as TA with IBDU. Colonic inflammation was initially discontinuous and stronger in the proximal colon, atypical of UC. However, over 10-year observation, the distribution of colonic inflammation varied and became UC-like. Variations in TA-related colonic inflammations over time have been unclear. Our long-term observation might help clarify the details of TA-related colonic inflammation.

Abstract Aim We recently reported that lipoprotein lipase (LPL)‐mediated free cholesterol (FC) accumulation in hepatic stellate cells (HSCs) augmented liver fibrosis in non‐alcoholic steatohepatitis (NASH). The aim of the present study was to explore the role of angiopoietin‐like protein 4 (Angptl4), an LPL inhibitor, in the pathogenesis of liver fibrosis in NASH. Methods Angptl4‐deficient or wild‐type mice were used to investigate the role of Angptl4 in the pathogenesis of NASH induced by feeding a methionine‐ and choline‐deficient diet. We also examined the effect of Angptl4 on FC accumulation in HSCs, and the subsequent activation of HSCs, using Angptl4‐deficient HSCs. Results In the NASH model, Angptl4‐deficient mice had significantly aggravated liver fibrosis and activated HSCs without enhancement of hepatocellular injury, liver inflammation, or liver angiogenesis. FC levels were significantly higher in HSCs from Angptl4‐deficient mice than in those from wild‐type mice. Treatment with Angptl4 reversed low‐density lipoprotein‐induced FC accumulation in HSCs through the inhibition of LPL. The Angptl4 deficiency‐induced FC accumulation in HSCs suppressed HSC expression of the transforming growth factor‐ β (TGF‐ ß ) pseudoreceptor, bone morphogenetic protein, and activin membrane‐bound inhibitor, and sensitized HSCs to TGF‐ β ‐induced activation in vivo and in vitro. Conclusions Angptl4 plays an important role in the pathogenesis of FC accumulation in HSCs. In addition, regulation of FC levels in HSCs by Angptl4 plays a critical role in the pathogenesis of liver fibrosis in NASH. Thus, Angptl4 could represent a novel therapeutic option for NASH.

Abstract Introduction Innate lymphoid cells (ILCs) are abundant in the intestinal mucosa, forming boundaries externally. Herein, ILCs were directly obtained from intestinal lymph using a lymph fistula rat model and analyzed under physiological and pathological conditions. Methods Thoracic duct (TD) lymphocytes were collected by cannulation with/without preceded mesenteric lymphadenectomy, which were comparable to lymphocytes flowing through mesenteric lymphatic vessels (MLVs) or TD, respectively. The collected ILCs were classified according to gene transcription factors and analyzed by flow cytometry. The effect of IL‐25 or indomethacin was studied. Results The proportion of total ILCs in the MLVs (MLV‐ILCs) was significantly higher than that in TD (TD‐ILCs, 0.01% vs. 0.003%, respectively). Physiologically, there were several significant differences in the MLV‐ILCs compared with TD‐ILCs, including the proportion of ILC2 (42.3% vs. 70.9%) and ILC3 (33.3% vs. 13.8%), and the proportion of α4‐integrin‐positive cells (36.8% vs. 0.3%). IL‐25 significantly increased the proportion of MLV‐ILC2 after 3 days. Indomethacin‐induced intestinal injury increased the proportion of MLV‐ILC3 in the early phase within 12 h. Conclusion Intestinal ILCs were found to migrate through MLVs. The altered mobilization of MLV‐ILCs after stimuli suggests that ILCs play an important role in regulating the immune responses at the secondary lymph nodes.

Nonsteroidal anti-inflammatory drug (NSAID)-induced enteropathy, the mechanism of which is involved in oxidative stress, can be lethal due to hemorrhage. Thus, we aimed to investigate the effect of hydrogen-rich water (HRW), in terms of oxidative stress, on intestinal mucosal damage as well as changes in the gut microbiome and the short-chain fatty acids (SCFAs) content in feces.Hydrogen-rich water was orally administered for 5 days to investigate the effectiveness of indomethacin-induced enteropathy in mice. Small intestinal damage and luminal reactive oxygen species (ROS) were evaluated to investigate the ameliorating effects of hydrogen. Then, components of the gut microbiome were analyzed; fecal microbiota transplantation (FMT) was performed using the cecal contents obtained from mice drinking HRW. The cecal contents were analyzed for the SCFAs content. Finally, cells from the macrophage cell line RAW264 were co-cultured with the supernatants of cecal contents.Hydrogen-rich water significantly ameliorated IND-induced enteropathy histologically and reduced the expression of IND-induced inflammatory cytokines. Microscopic evaluation revealed that luminal ROS was significantly reduced and that HRW did not change the gut microbiota; however, FMT from HRW-treated animals ameliorated IND-induced enteropathy. The SCFA content in the cecal contents of HRW-treated animals was significantly higher than that in control animals. The supernatant had significantly increased interleukin-10 expression in RAW264 cells in vitro.Hydrogen-rich water ameliorated NSAID-induced enteropathy, not only via direct antioxidant effects but also via anti-inflammatory effects by increasing luminal SCFAs. These results suggest that hydrogen may have therapeutic potential in small intestinal inflammatory diseases.

Mild blast-induced traumatic brain injury (bTBI) induces various gut symptoms resembling human irritable bowel syndrome (IBS) as one of mental and behavioral disorders. However, the underlying mechanisms remain unclear. We investigated whether the extremely localized brain impact extracranially induced by laser-induced shock wave (LISW) evoked IBS-like phenomenon including visceral hypersensitivity and intestinal hyperpermeability in rats.The rats were subjected to LISW on the scalp to shock the entire brain. Visceral hypersensitivity was evaluated by the threshold pressure of abdominal withdrawal reflex (AWR) using a colorectal distension test. Permeability was evaluated by the concentration of penetrating FITC-dextran from intestine and the mRNA expression levels of tight junction family proteins. Involvement of corticotropin-releasing factor receptor (CRFR) 1 and 2 was examined by evaluating mRNA expression and modulating CRFR function with agonist, recombinant CRF (10 μg/kg), and antagonist, astressin (33 μg/kg). High-throughput sequencing of the gut microbiota was performed by MiSeqIII instrument and QIIME tool.The thresholds of the AWR were significantly lowered after LISW. Permeability was increased in small intestine by LISW along with decreased expression of tight junction ZO-1. LISW significantly increased CRFR1 expression and decreased CRFR2 expression. Visceral hypersensitivity was significantly aggravated by CRFR agonist and suppressed by CRFR antagonist. The α- and β-diversity of the fecal microbiota was altered after LISW.LISW provoked visceral hypersensitivity, small intestinal hyperpermeability, altered expression of CRFRs and changes in the microbiota, suggesting that genuine bTBI caused by LISW can induce a pathophysiology comparable to that of human IBS.

Abstract Background and Aim The small intestine plays a central role in gut immunity, and enhanced lymphocyte migration is involved in the pathophysiology of various enteropathy. Bile acid (BA) is closely related to lipid metabolism and gut microbiota and essential for gut homeostasis. However, the effects of BA on gut immunity have not been studied in detail, especially on the small intestine and lymphocyte migration. Therefore, we aimed to investigate the effect of BA on small intestinal lymphocyte microcirculation. Methods The effect of deoxycholic acid (DCA), taurocholic acid (tCA), or cholic acid (CA) on the indomethacin (IND)‐induced small intestinal enteropathy in mice was investigated. Lymphocyte movements were evaluated after exposure to BA using intravital microscopy. The effects of BA on surface expression of adhesion molecules on the vascular endothelium and lymphocytes through BA receptors were examined in vitro . Results IND‐induced small intestinal enteropathy was histologically aggravated by DCA treatment alone. The expression of adhesion molecules ICAM‐1 and VCAM‐1 was significantly enhanced by DCA. Exposure to DCA increased lymphocyte adhesion in the microvessels of the ileum, which was partially blocked by anti‐α4β1 integrin antibody in vivo . The expression of ICAM‐1 and VCAM‐1 was significantly enhanced by DCA in vitro , which was partially suppressed by the sphingosine‐1‐phosphate receptor 2 (S1PR2) antagonist. The S1PR2 antagonist significantly ameliorated IND‐induced and DCA‐exaggerated small intestinal injury. Conclusion DCA exacerbated IND‐induced small intestinal enteropathy. DCA directly acts on the vascular endothelium and enhances the expression levels of adhesion molecules partially via S1PR2, leading to enhanced small intestinal lymphocyte migration.

We herein report a 44-year-old man suffering from systemic edema due to protein-losing enteropathy (PLE) with superior mesenteric vein (SMV) obstruction and development of collateral veins, which subsequently proved to be a chronic result of thrombosis and a complication of Crohn's disease (CD). PLE was supposedly induced by both intestinal erosion and thrombosis-related lymphangiectasia, which was histologically proven in his surgically-resected ileal stenosis. Elemental diet and anti-TNFα agent improved his hypoalbuminemia after surgery. The rarity of the simultaneous coexistence of SMV obstruction and PLE and the precedence of these complications over typical abdominal symptoms of CD made the clinical course complex.

The study of the impact of environmental factors during pregnancy on fetal development has so far been focused primarily on those negatively affecting human health; however, little is known about the effects of probiotic treatment during pregnancy on inflammatory bowel diseases (IBD). In this study, we investigated whether oral administration of heat-killed probiotics isolated from fermented foods decreased the vulnerability of offspring to IBD.Probiotics were administered to the pregnant mice until the birth of pups, after which the parent mice were maintained with autoclaved water. Partial pups were evaluated for dextran sodium sulfate-induced colitis. The influence of CD11c+ CD103+ dendritic cells (DCs) and regulatory T cells (Tregs) in mesenteric lymph nodes of parent mice and their pups was analyzed.Oral administration of heat-killed probiotics to pregnant dams significantly decreased inflammation induced by dextran sodium sulfate in pups. Probiotic treatment increased the number of CD103+ DCs, and the expression of β8-integrin in CD103+ DCs and Tregs in mesenteric lymph nodes, not only in dams themselves but also in their offspring.Oral administration of probiotics during gestation induced transgenerational immunomodulatory effects on the gut-associated immune system and resilience to experimental colitis in the offspring. Our results suggest that consumption of fermented foods during pregnancy can be effective in preventing inflammatory diseases such as IBD beyond generation.

We herein present the case of an immunocompetent 63-year-old man who had previously undergone resection of Crohn's disease (CD)-related small intestinal obstruction more than 30 years ago. He had not been receiving any medication for many years, but had recently started to suffer from ileus. A stenosed site of ileo-cecal anastomosis was identified and therefore was surgically resected, which was diagnosed as CD with small intestinal extramedullary plasmacytoma (EMP). The subsequent progression of CD was successfully controlled by anti-TNFα agents without any recurrence of EMP for over 3 years, implying the clinical benefit and safety of the biological therapy. This was the first known case of a patient who received anti-TNFα agents after a resection of small intestinal EMP accompanied with CD.

We herein describe a 69-year-old man suffering from chronic diarrhea caused by lansoprazole (LPZ)-induced collagenous colitis (CC) accompanied with protein-losing enteropathy (PLE), diagnosed by increased fecal alpha-1 antitrypsin clearance and the findings of leakage from the descending colon to the sigmoid colon on scintigraphy. MR enterocolonography (MREC) was also performed for differentiating digestive diseases, and inflamed findings were observed around the same portion as those on scintigraphy, suggesting that this region was responsible for protein loss in this case. The MREC findings improved after the cessation of LPZ, and hypoalbuminemia also improved simultaneously. This case suggests that MREC may be a new and useful diagnostic tool for CC with PLE.

Immunoglobulin (Ig)G4-related disease (IgG4-RD) is a systemic condition associated with fibroinflammatory lesions and is characterized by elevated serum IgG4 levels and IgG4-positive cell infiltration into the affected tissues. It has been reported that IgG4-RD affects a variety of organs but uncommonly affects the gastrointestinal tract. In particular, there are few cases of lesions in the small intestine, except for sclerosing mesenteritis, which were mostly diagnosed from surgical specimens. Herein, we describe the case of a 70-year-old man who initially presented with abdominal pain, headache, later cognitive decline, and gait disturbance caused by IgG4-RD. Colonoscopy revealed irregular ulcers in the terminal ileum, and computed tomography of the head showed hypertrophic pachymeningitis. Numerous IgG4-positive cells were detected in the ileal and dural biopsies. We diagnosed the patient with IgG4-RD and started steroid pulse therapy. After initiation of treatment, the symptoms quickly improved. The patient was discharged from the hospital after starting oral prednisolone treatment (30 mg). The dosage was gradually reduced to 10 mg. A follow-up colonoscopy revealed scarring of the ileal ulcers. This case may provide valuable information regarding the endoscopic findings of small intestinal lesions in IgG4-RD.

Abstract Background and Aim The functions of basophils have not been elucidated until recently because of their rarity. However, with recent developments in basophil‐specific antibodies and basophil‐deficient animals, the roles of basophils in various diseases related to chronic inflammation have been clarified. In this study, we aimed to investigate the roles of basophils in human ulcerative colitis (UC) and oxazolone (OXA) colitis using genetically engineered Mcpt8 DTR mice. Methods Immunohistochemical staining of human colon specimens was performed to examine the involvement of basophils in the pathogenesis of UC. We examined the correlation between the number of infiltrating basophils and the UC endoscopic index of severity (UCEIS), Mayo score, and Matts score. We also examined the correlation between eosinophil count and basophil infiltration. In murine experiments, we examined whether basophil infiltration was involved in OXA‐induced colitis and whether basophil depletion improved inflammation in Mcpt8 DTR mice. Results Colonic basophil infiltration was significantly increased in patients with UC. There were significant correlations between UCEIS, Mayo score, Matts score, and the number of infiltrating basophils. In murine OXA‐induced colitis, a significant increase in basophil infiltration was observed. When basophils were depleted by diphtheria toxin in Mcpt8 DTR mice, inflammation improved significantly and mRNA expression of some proinflammatory cytokines, including Tnf‐α and Ifn‐γ decreased significantly. Conclusion Basophil infiltration correlated with endoscopic, clinical, and pathological scores in human UC independently of eosinophil infiltration, and depletion of basophils ameliorated mucosal inflammation in murine OXA‐induced colitis, collectively suggesting that basophils exert a proinflammatory role in chronic intestinal inflammation such as UC.

could be identified by MT stain over time.Fibrosis was not significantly different in MT stain until 24 hours, but significantly increased until 7 days thereafter.The fraction of the Caspase-3 area was calculated from the sum of the casepase-3 and TUNEL staining areas, which was 11.96 times higher at 200V compared to the control.At last, caspase-3 protein was confirmed by western blot.The most caspase-3 protein was identified in the 200V group.Conclusion: The optimal treatment range was 200V to 400V and the energy with the highest percentage of apoptosis was 200V, which is corresponding to electrical energy 1000V/cm.

A 70-year-old asymptomatic man presented with a growing submucosal tumor in the duodenal bulb; and despite two EUS-FNA procedures, a diagnosis could not be confirmed. Since the mass was stalked and enlarged to 40 mm in size, the patient underwent endoscopic resection with grasping scissor forceps, while traction was applied for diagnostic treatment, considering the risk of gastrointestinal obstruction. The pathological diagnosis was Brunner's gland hyperplasia, and the patient was followed up with annual upper gastrointestinal endoscopy. Here, we report a case of endoscopic resection of a large duodenal submucosal tumor using a traction device.

This study is about the octagonal building of GYEONGJU NAJONG, the remains of a ritual facility of ancient Shilla Dynasty. The purpose of this study is to examine the column arrangement of the octagonal building, and reveal the determinant factors for column arrangement by reviewing the sequential processes in building the facility.The results are as follows : the octagonal building was designed with special structure that internal structure is rotated against external column arrangement. Difference in the number of columns between inside and outside was caused by restrictions on the scale of the inner and outer circumference. And it induced rotation of the internal structure for the external column array. The circumference of internal and external column of the octagonal building was defined to include the core portion of predecessor facility, consisting of the central pit and the six pits.

A 67-year-old woman who had been treated for intraductal papillary mucinous carcinoma with bile duct jejunum anastomosis was admitted due to fever. She had undergone three sessions of repeated transcatheter arterial chemoembolizations (TACE) and two sessions of repeated radiofrequency ablation for multiple hepatocellular carcinomas at our hospital. We diagnosed biloma with an abdominal CT scan. We performed endoscopic retrograde cholangiopancreatography (ERCP) and detained an endscopic indwelling biloma drainage tube. The biloma rapidly decreased. ERCP and EBD were useful in this case that had difficulty in the endscopic approach.

A 70-year-old man with past medical history of left renal cell carcinoma status post nephrectomy in 1988 presented to our department, complaining of melena and anemia. He underwent partial gastrectomy for gastric metastasis in 2007, followed by pancreatectomy for pancreatic metastasis, by radiofrequency ablation for right renal metastasis (2009) , and by molecular target therapy for multiple liver metastasis and retroperitoneal metastasis (2012) . He also had an evidence of right adrenal metastasis in 2013. On gastrointestinal endoscopy, a protruding lesion covered with white moss was found on greater curvature of the upper part of gastric body (color 1, 2, 3) . The lesion showed bleeding diathesis during the procedure. The pathological findings of the lesion were consistent with the gastric metastasis of renal cell carcinoma. Although the patient’s initial symptoms, melena and anemia, were disappeared spontaneously after the biopsy, the patient decided to take palliative care including the deep sedation. Here, we report a case of gastric metastasis of renal cell carcinoma, which presented with gastrointestinal hemorrhage.

A 66-year-old man experienced epigastric pain after massive vomiting. He was diagnosed with spontaneous perforation of the esophagus. Laparoscopic transhiatal repair was immediately performed. After surgery, esophageal-mediastinal fistula resulting from the anastomotic insufficiency remained and didn’t improve with conservative treatment. We attempted to close the fistula with a polyglycolic acid sheet and fibrin glue via endoscopy. First, a polyglycolic acid sheet was packed in the fistula. Then, we sprayed fibrin glue on the sheet. Finally, we used endoscopic clips to prevent from falling off the sheet. As a result, leakage stopped completely and the endoscopy confirmed closure of the fistula. We report endoscopic closure of the remaining esophageal fistula after surgical repair with literature reviews.

Lipopolysaccharides (LPS) entry from the intestinal lumen to the circulation is associated with systemic inflammation. We have found that LPS is transcellularly transported to portal vein during long-chain fatty acid (LCFA) exposure via CD36- and lipid raft-mediated pathways in rats, suggesting the involvement of caveolae-mediated endocytosis. We thus examined LPS transport in caveolin-1 (Cav1) knockout (KO) murine jejunum. FITC-LPS was applied to the mucosal bath of Ussing chambered muscle-stripped jejunal mucosa-submucosa preparations of Cav1 KO and wild type (WT) mice. Serosal appearance of FITC-LPS was measured with or without luminal application of oleic acid (OA, 30 mM) with taurocholic acid (TCA, 0.1 mM), or medium-chain fatty acid sodium caprate (C10, 10 mM). Luminal application of OA/TCA increased FITC-LPS m-to-s transport in WT jejunum, inhibited by the CD36 inhibitor sulfosuccinimidyl oleate (SSO, 0.1 mM) or the lipid raft inhibitor methyl-β-cyclodextrin (MβCD, 0.1 mM), not by the clathrin inhibitor chlorpromazine (0.1 mM) or Pitstop2 (30 µM), suggesting that LCFA-induced LPS transport is mediated by caveolae-mediated endocytosis, not by clathrin-mediated endocytosis. In contrast, OA/TCA-induced FITC-LPS transport was abolished in Cav1 KO jejunum. Nevertheless, luminal application of C10 increased FITC-LPS transport both in WT and Cav1 KO jejuna without transepithelial electrical resistance changes, suggesting that C10 enhances transcellular LPS transport via caveolin-independent endocytosis in the jejunum. These results suggest that LPS transport during LCFA exposure is mediated by Cav1-mediated endocytosis, whereas MCFA-induced LPS transport is likely via clathrin-mediated endocytosis. Modulation of epithelial endocytosis may be a new therapeutic target for LPS-associated diseases mediated by dietary fatty acids, including metabolic syndrome.

Medium-chain triglycerides (MCT) in diet improves glucose tolerance and insulin sensitivity, compared to long-chain triglycerides (LCT). However, the direct effects of medium-chain fatty acids (MCFA) on glucose transport have not been studied. Luminal glucose is absorbed in the small intestine through the electrogenic sodium-dependent glucose transporter SGLT1 and electroneutral glucose transporter GLUT2. We thus examined the effects of luminal MCFA on electrogenic SGLT1 activity and glucose absorption in mice. SGLT1 activity was assessed in the Ussing chambered, muscle-stripped jejunal mucosa by measuring phlorizin (Phz)-sensitive short-circuit current (Isc ) evoked by luminal addition of glucose (10 mM). Sodium propionate (C3), caproate (C6), caprylate (C8), caprate (C10), laurate (C12) and myristate (C14) were luminally added before or after glucose application. Furthermore, the effects of C10 on glucose absorption were assessed by oral glucose tolerance test (OGTT, glucose 2g/kg body weight, ig) with or without C10 treatment (1.94g/kg, ig) in conscious mice. Moreover, the portal venous (PV) blood glucose levels were measured though the PV cannulation, followed by intraduodenal bolus injection of glucose solution (100 mM) with or without C10 (10 mM) in the anesthetized mice. Luminal pretreatment of C10 and C12 dose-dependently inhibited the glucose-induced Isc increase at IC50 = 1.02 mM and 1.06 mM, respectively. C8 also inhibited the glucose-induced Isc increase, but less effective (IC50 8.4 mM), whereas C3, C6 and C14 had little effect. C10 (10 mM) also inhibited methyl α-D-glucopyranoside (αMG)-induced Isc increase. Post-treatment of C10 and C12 (10 mM) rapidly reduced the increased Isc by glucose, mimicking the inhibitory effect of Phz (1 mM). OGTT showed that glucose levels reached the peak at 30 min, while C10 pretreatment remarkably inhibited glucose increase. Furthermore, intraduodenal injection of glucose solution increased the PV glucose levels at 10 - 20 min, while the addition of C10 in the glucose solution abolished the response. These data suggest that C10 and C12 act as the SGLT1 inhibitor. Since MCT releases C10 and C12 via digestion by pancreatic lipase, slower than glucose release by maltase/sucrase, free C10/C12 solution may be more effective therapeutics for the treatment of type 2 diabetes.

The introduction of the western diet has been proposed as an explanation for the increase in inflammatory bowel disease (IBD) incidence. Among them, greater consumption of fat is known to increase risk of IBD. We have reported that dietary fat augmented intestinal immune system by increasing lymphocyte migration to the intestinal microvessels by using intravital microscope system in the animal models of IBD. High fat diet increases secondary bile acid, especially deoxycholate (DCA), which is reported to be involved in the exacerbation of IBD along with direct effect of fat. Each bile acid distinctively affects gut immunity and its mechanism remains to be clarified. Recently, aberrant migration of lymphocytes to intestinal mucosa has been regarded one of critical pathogenesis of IBD. We aimed to investigate the effect of several kinds of bile acid on gut immunity in the view point of intestinal microcirculation. (1) Effect of bile acid exposure on intestinal mucosa: Thoracic duct lymphocytes (TDL) were collected from the thoracic duct of donor rats. We intravenously injected CFSE-fluorescence labelled TDL into recipient rats, and migration in intestinal mucosa was observed by a confocal microscope to evaluate the TDL migration. In some recipient rats, bile acids were injected into ligated ileum at both ends to evaluate the direct effect on intestinal mucosa. Tauro Colic Acid Natrium (tauro-CANa, 4 mM) or DCA (4 mM), PBS were injected into the intestinal lumen. PBS was used as a control sample. Mucosal vascular addressin cell adhesion molecule 1 (MAdCAM-1) neutralising antibody was administered intravenously in some rats. Mucosal damages were measured histologically. (2) Effect of bile acid exposure on TDL: TDL were cultured at 4°C for 2 h with above-mentioned bile acids. Expression levels of L-selectin and α4 integrin in the obtained lymphocytes were examined by flowcytometry. (1) A small number of lymphocytes adhered to intestinal microvessels in control group. There was no change in TDL adhesion in the Tauro-CANa exposed group. TDL adhesion increased in the DCA exposed group. Increased lymphocyte adhesion by DCA was partially blocked by neutralising antibody of MAdCAM-1. Histologically, a part of intestinal mucosa was damaged by DCA, and inflammatory cell infiltration was observed in the mucosa. (2) Expressions of α4 integrin and L-selectin on TDL did not alter with or without addition of bile acids. DCA caused injury of ileal mucosal epithelium and increased lymphocyte adhesion to the vascular endothelium in the ileal mucosa, suggesting that the gastrointestinal immunity could be altered by some bile acids via increase in expression of adhesion molecules on microvessels.

Background: Human stem cell derived enteroid monolayers have become a widely used model to study intestinal physiology and pathophysiology.However, this model lacks aspects of normal intestine including other cell types and exposure to mechanical forces including luminal and blood flow (shear force) and peristalsis.Here, we determine the effects of physical forces applied to human differentiated jejunal enteroid monolayers in a diarrheal disease model developed in microengineered Intestine-Chip.We compared the effects of ST on cGMP production under static conditions, apical (AP) and basolateral (BL) flow, and flow plus cyclic stretch.We previously established the static ST-enteroid model as having: a) Limited increase in intracellular cGMP content.b) cGMP secretion across the BL>AP membrane.c) Extracellular cGMP transport dependent on MRPs.Methods: Differentiated human jejunal monolayers in Emulate PDMS "Intestine Chips" were exposed apically and basolaterally to enteroid media lacking WNT3A and R-SPONDIN-1 but, containing the PDE inhibitors IBMX (1mM) and vardenafil (1uM).On 5 th day post plating, control solution or ST (1nM) (concentration present in this model with ETEC exposure) were incubated on the AP for 6 hours under static or AP plus BL Flow (60ul/h) or Flow plus Stretch (0.15 Hz, 10% strain) conditions.AP, BL and intracellular cGMP were measured by ELISA.MRP 4 and 5 expressions was measured by qRT-PCR.Results: 1) Under basal conditions (no ST), 6h application of flow and flow plus stretch did not alter basal cGMP content but similarly increased AP and BL cGMP secretion.2) ST increased intracellular cGMP content and AP and BL cGMP secretion in all conditions.3) Flow and flow plus stretch similarly increased the ST-stimulated intracellular cGMP content and BL cGMP secretion> static conditions but did not alter apical secretion.4) MRP4, but not MRP5 mRNA was increased by flow and flow plus stretch compared to static conditions in both basal and ST conditions.Conclusions: 1) Mechanical forces mimicking luminal and blood flow and cyclic stretch altered the effect of ST to increase cGMP in human enteroids.2) Flow and flow plus stretch increased mRNA of MRP4 but not MRP5.3) Under basal conditions, flow and flow plus stretch similarly stimulated AP and BL cGMP secretion, an effect expected with the increased MRP4.ST addition also increased intracellular cGMP with flow and flow plus stretch>static.4) Flow is the major physical force responsible for changes in regulation of MRP4, increase in intracellular and BL cGMP secretion under basal and ST stimulated conditions; only the increase in MRP4 mRNA after ST appeared to depend more on stretch added to flow.In summary, application of physical forces alters the ST model of ETEC diarrhea and should be evaluated for effects on intestinal physiology and pathophysiology.

involved in chemokine production, cytokine-cytokine receptor interaction and regulation of actin and cytoskeleton.In particular, several fibrosis-related genes (Acta2, Axl, Nexn, etc) were significantly increased by OGT knockdown.Promoter sequence analysis of these genes indicated SRF as a common transcription factor.We further demonstrated that OGT repressed the activity of SRF to induce α-SMA transcription.Mutations of specific O-GlcNAcylation sites on SRF increased its transcriptional activity.Addtionally, OGT suppressed the promoter activity of other genes related to HSC activation via SRF.Finally, we found that Thiamet-G, a potent O-GlcNAcase inhibitor, raised the overall O-GlcNAcylation level in the liver, reduced expression of fibrogenic markers, and ameliorated CCl 4 -induced liver fibrosis.Conclusion: Our results reveal that OGT acts as a crucial negative regulator of HSC activation.Targeting O-GlcNAc signaling represents a novel potential strategy to prevent and treat liver fibrosis.

histological, and gene expression analyses.The expression levels of the ER-stress related genes Jun proto-oncogene (Jun), Atf3, Nupr1, and C/EBP homologous protein (Chop) were measured in liver tissue.Apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining.Results Control mice demonstrated hepatic steatosis alone without apparent fibrosis.On the other hand, FLS-ob/ob mice showed severe steatohepatitis at both 24 and 36 weeks of age and severe fibrosis at both 36 and 48 weeks of age.The expression levels of Atf3, Nupr-1, and Jun significantly increased from 24 to 48 weeks of age in FLS-ob/ob mice compared with control mice.The expression level of Chop was already high in FLS mice and maintained similar levels in FLS-ob/ob mice; the expression level was consistent with the percentage of TUNEL-positive cells.Conclusion The ATF3/ NUPR1 axis plays a pivotal role in NASH progression in association with Jun and Chop and appears to induce apoptosis from early steatosis in the NASH model mice.

surprise, in contrast to the muscle from other sites, the circular muscle of the narrowed colon was rich in LPS and showed a significant reduction of MYPT1 protein.The affected smooth muscle displayed a spastic contraction in terms of an enhanced sustained force tension and a resistant relaxation to PKC/ROCK inhibitors.Our collective observations suggest that LPS-induced protein degradation may serve as an essential mechanism regulating MYPT1 protein under pathological condition.Such a regulation may contribute to the colonic obstruction of HD disease.

In our resent paper, we explained the planning method of the column interval dimensions of Horyuji Kondo (the main hall of Horyuji temple) by adopting the mediaeval period's 'shiwari-sei' as a clue. However, what seems to be lacking is the consideration to the Pagoda in Asuka period. This thesis deals with the method for designing the standard unit of the column interval of Horyuji Gojyu-no-tou (Pagoda of Horyuji temple) and Hokkiji Pagoda. These Pagodas have the same measure and the method founded on the primitive 'shiwari-sei' as that of Horyuji Kondo and Chumon.

with disease severity and infliximab requirement.Patients with higher Nancy scores (3/4) were more likely to require infliximab (p=0.03).Conclusions Clinical assessment with CTCAE doesn't accurately reflect irAE colitis severity thus negatively impacting timely treatment decisions.Our data support endoscopic scoring of colitis severity and demonstrate the potential prognostic utility of objective histologic scoring.

A 69-year-old man had been intermittently experiencing abdominal pain from his 30s and was diagnosed with colonic diverticulitis. He further experienced right lower abdominal pain and received treatment. However, his condition did not improve, and he was referred to the National Defense Medical College Hospital. His abdominal pain episodes continued even after treatment for few weeks;subsequently, familial Mediterranean fever (FMF) was suspected based on the clinical course because of elevated inflammatory responses, although his body temperature was ≤38°C. After administrating colchicine as a diagnostic treatment, the repeated abdominal pain disappeared. Considering the other findings and genetic examination that showed the representative gene mutation of MEFV (M694I), he was diagnosed with FMF. This case indicates that high body temperature, one of the primary diagnostic criteria of FMF, is sometimes not evident in elderly patients, thereby causing potential misdiagnosis in some elderly patients with FMF.