[A rare case of intraductal tumor of the pancreas in which an intraductal tubulopapillary neoplasm was mixed with a widely spreading gastric-type intraductal papillary-mucinous neoplasm].

A 70-year-old man receiving treatment for diabetes mellitus presented with a cystic mass in the border area of the pancreatic body and tail on plain computed tomography (CT) due to impaired glucose intolerance. Contrast-enhanced CT showed a faint hyperattenuated nodular mass extending from the dilated main pancreatic duct (MPD) to the branch duct. Endoscopic retrograde cholangiopancreatography revealed a mildly dilated orifice of the papilla of Vater and MPD stenosis with entire upstream and immediate downstream dilatations. The patient underwent distal pancreatectomy due to the suspicion of mixed-type intraductal papillary-mucinous carcinoma. A pathological examination showed an intraductal solid-nodular mass measuring 25mm in length, consisting of two types of neoplasms. One showed tubulopapillary growth with entirely high-grade (HG) atypical cuboidal epithelium, in which immunohistochemical examinations were positive for MUC6 but negative for human gastric mucin (HGM), MUC1, MUC2, and MUC5AC, fitting the concept of intraductal tubulopapillary neoplasm (ITPN). The other showed the same growth of low-grade (LG) atypical columnar cells positive for HGM and MUC5AC and negative for MUC1 and MUC2, which corresponded to gastric-type intraductal papillary-mucinous neoplasm (IPMN) -LG. The tumor had not invaded the duct walls, and no metastatic lymph nodes were observed. The ITPN was adjacent to the IPMN mainly composed of tubular glands mimicking pyloric glands with LG dysplasia that corresponded to the so-called IPMN-pyloric gland variant. Moreover, the proliferation of low-papillary gastric-type IPMN spread around the intraductal tumors. Consequently, the patient was diagnosed with an intraductal tubular neoplasm comprising a noninvasive ITPN and gastric-type IPMN-LG. ITPN is a recently identified intraductal neoplasm of the pancreas proposed by Yamaguchi et al. and is distinguished by intraductal tubulopapillary growth with HG cellular atypia without overt mucin production, in contrast to IPMN. To date, no cases of intraductal nodular tumors comprising ITPN and IPMN have been reported. We report this original case with imaging and pathological observations and discuss potential processes via which ITPN and IPMN may arise adjacent to each other in the same pancreatic duct.

Cryoglobulinemia vasculitis associated with adult-onset Still's disease.

Key Clinical Message: The present case indicates that cryoglobulinemia vasculitis should be considered in the differential diagnosis of purpura in patients with adult-onset Still's disease (AOSD). Abstract: The presence of purpura is suggested in adult-onset Still's disease (AOSD) hematological complications of hemophagocytic syndrome, disseminated intravascular coagulation, or thrombotic microangiopathy. We herein report a case of AOSD complicated by cryoglobulinemia vasculitis presenting with purpura.

Glioblastoma/high-grade glioma with a primitive neuronal component including rhabdoid differentiation that was difficult to diagnose: A case report.

Glioblastoma with a primitive neuronal component (GBM-PNC) is a rare subtype. In this case, GBM-PNC was difficult to diagnose conclusively because the specimen consisted of only a few high-grade glioma components. A 73-year-old woman presented with sensory aphasia and minor right-sided hemiplegia. Imaging revealed a neoplastic lesion with a maximum diameter of approximately 5 cm in the left frontal lobe for which surgery was performed. Histologically, most atypical cells were immature components with high nuclear-cytoplasmic ratios and immunopositive for neuroendocrine markers. Minor components of atypical glial cells were found at tumor margins. Rhabdoid cells were observed in undifferentiated components. Immunostaining was positive for glial fibrillary acidic protein (GFAP), nestin, and Olig2 in both undifferentiated and atypical glial cells. The major undifferentiated components showed significantly low GFAP, nestin, and Olig2 expression levels within the foci of the undifferentiated components, in contrast to the atypical glial component, neurofilaments and synaptophysin were immunopositive for undifferentiated components. Rhabdoid cells were immunopositive for myogenin, desmin, and HHF35, suggesting their differentiation into striated muscles. This was a particularly rare case because rhabdoid differentiation was observed in PNC.

A Lethal Combination: Legionnaires' Disease Complicated by Rhabdomyolysis, Acute Kidney Injury, and Non-Occlusive Mesenteric Ischemia.

BACKGROUND Legionnaires' disease is one of the most common types of community-acquired pneumonia. It can cause acute kidney injury and also occasionally become severe enough to require continuous renal replacement therapy (CRRT). Non-occlusive mesenteric ischemia (NOMI) is a condition characterized by ischemia and necrosis of the intestinal tract without organic obstruction of the mesenteric vessels and is known to have a high mortality rate. CASE REPORT A 72-year-old man with fatigue and dyspnea was diagnosed with Legionnaires' disease after a positive result in the Legionella urinary antigen test pneumonia confirmed by chest radiography and computed tomography. He developed acute kidney injury, with anuria, rhabdomyolysis, septic shock, respiratory failure, and metabolic acidosis. We initiated treatment with antibiotics, catecholamines, mechanical ventilation, CRRT, steroid therapy, and endotoxin absorption therapy in the Intensive Care Unit. Despite ongoing CRRT, metabolic acidosis did not improve. The patient was unresponsive to treatment and died 5 days after admission. The autopsy revealed myoglobin nephropathy, multiple organ failure, and NOMI. CONCLUSIONS We report a fatal case of Legionnaires' disease complicated by rhabdomyolysis, acute kidney injury, myoglobin cast nephropathy, and NOMI. Legionella pneumonia complicated by acute kidney injury is associated with a high mortality rate. In the present case, this may have been further exacerbated by the complication of NOMI. In our clinical practice, CRRT is a treatment option for septic shock complicated by acute kidney injury. Thus, it is crucial to suspect the presence of NOMI when persistent metabolic acidosis is observed, despite continuous CRRT treatment.

Efficacy of Rectal Systemic Administration of Mesenchymal Stem Cells to Injury Sites via the CXCL12/CXCR4 Axis to Promote Regeneration in a Rabbit Skeletal Muscle Injury Model.

Mesenchymal stem cells (MSCs) have been transplanted directly into lesions or injected intravenously. The administration of MSCs using these delivery methods requires specialized knowledge, techniques, and facilities. Here, we describe intrarectal systemic administration of MSCs, a simple, non-invasive route for homing to the injury sites to promote the regeneration of skeletal muscle injuries. Using a cardiotoxin (CTX)-induced rabbit skeletal muscle injury model, homing to the site of muscle injury was confirmed by intrarectal administration of MSCs; the time required for homing after intrarectal administration was approximately 5 days. In addition, the C-X-C chemokine ligand 12 (CXCL12)/C-X-C chemokine receptor-4 (CXCR4) axis was found to be involved in the homing process. Histopathological examinations showed that skeletal muscle regeneration was promoted in the MSCs-administered group compared to the CTX-only group. Myosin heavy polypeptide 3 (Myh3) expression, an indicator of early muscle regeneration, was detected earlier in the intrarectal MSCs group compared to the CTX-only group. These findings indicate that intrarectal administration of MSCs is effective in homing to the injured area, where they promote injury repair. Since intrarectal administration is a simple and non-invasive delivery route, these findings may be valuable in future research on stem cell therapy.

Solitary juvenile polyp of the rectum with intramucosal adenocarcinoma.

We describe a case of solitary juvenile polyp of the rectum with intramucosal adenocarcinoma. A 55-year-old man presented to our hospital for evaluation after a positive fecal occult blood test. Colonoscopy revealed a pedunculated polyp of 25 mm in size which has an irregular shape and pale red color on the rectum. The polyp had a proliferation of blood vessels and an invisible surface pattern. Endoscopic mucosal resection was performed. Pathologically, it was diagnosed as a solitary juvenile polyp with intramucosal well-differentiated adenocarcinoma. When we encounter juvenile polyps, the possibility of malignancy should be taken into consideration for treatment.

[Carcinoma in situ arising in adenomyomatous hyperplasia of the ampulla of Vater: the first case report].

A man in his 70s was admitted to our hospital due to jaundice and upper abdominal pain. Laboratory findings indicated elevated serum hepatobiliary enzyme and amylase levels. Contrast-enhanced computed tomography revealed smooth wall thickening of the terminal bile duct (tBD) with a faintly enhanced inner line. ERCP revealed stenosis from the tBD to the ampulla of Vater (AV) with upstream dilatation. Intraductal ultrasound (IDUS) circumferentially revealed a thickened wall preserving a three-layered structure throughout the same region. Furthermore, a thick innermost hyperechoic layer was identified in the bile duct portion of the AV (Ab). Findings suggestive of adenocarcinoma were obtained from the tissue samples from the biliary stricture using biopsy forceps. Thus, pancreatoduodenectomy was performed. A pathological examination revealed a thickened AV wall spreading over the tBD with hyperplasia of the glands and smooth muscle fibers. In addition, low-grade biliary intraepithelial neoplasia (BilIN) was scattered throughout the lesion, and high-grade BilIN was partly observed in the peribiliary glands of the Ab. Based on these results, a diagnosis of carcinoma in situ arising in adenomyomatous hyperplasia (ADMH) of the AV was made. To date, there are no reports on ADMH-associated carcinoma of the BD or AV. We here report this original case with the IDUS findings, which are presumed to reflect the histologic features of ADMH showing ductal proliferation surrounded by smooth muscle fibers. Also, we discuss the process through which carcinoma arises from ADMH in AV.

The Possibility of IPC to Prevent Ischemic-Reperfusion Injury in Skeletal Muscle in a Rat.

Blood removal with air tourniquets for a long time induces muscle damage after reperfusion. Ischemic preconditioning (IPC) has a protective effect against ischemia-reperfusion injury in striated muscle and myocardium. However, the mechanism of action of IPC on skeletal muscle injury is unclear. Thus, this study aimed to investigate the effect of IPC in reducing skeletal muscle damage caused by ischemia-reperfusion injury. The hindlimbs of 6-month-old rats were wounded with air tourniquets at a carminative blood pressure of 300 mmHg on the thighs. Rats were divided into the IPC (-) group and the IPC (+) group. The vascular endothelial growth factor (VEGF), 8-hydroxyguanosine (8-OHdG), and cyclooxygenase 2 (COX-2) were investigated by protein levels. Quantitative analysis of apoptosis was performed using the TUNEL method. Compared with the IPC (-) group, the IPC (+) group retained the VEGF expression, and the COX-2 and 8-OHdG expressions were suppressed. The proportion of apoptosis cells decreased in the IPC (+) group compared with the IPC (-) group. IPC in skeletal muscles proliferated VEGF and suppressed inflammatory response and oxidative DNA damage. IPC has the potential to reduce muscle damage after ischemia-reperfusion.

Surgical implantation of human adipose derived stem cells attenuates experimentally induced hepatic fibrosis in rats.

BACKGROUND: Mesenchymal stem cells (MSCs) are multipotent stromal cells and could exert hepatoprotective effects against acute liver injury, steatohepatitis, and fibrogenesis. Here, we evaluated the effects of human adipose derived stem cells (hADSCs) to attenuate experimentally induced hepatic fibrosis and early cirrhosis in rats. METHODS: Hepatic fibrosis was induced by intraperitoneal injections of CCl4 (0.1 ml/100 g body weight) twice a week for 8 weeks. hADSCs were isolated and cultured on polyethylene discs coated with hydroxyapatite and 2 cm diameter disc was surgically implanted on the right lateral lobe of the liver. Discs implanted without hADSCs served as control. The animals were injected again with CCl4 once a week for another 8 weeks. All the animals were sacrificed at the end of 16th week. RESULTS: Serial administrations of CCl4 resulted in well developed fibrosis and early cirrhosis at 8th week which maintained until the 16th week. Animals treated with hADSC discs depicted over 50% decrease of collagen with significant increase in serum albumin and total protein levels. Immunohistochemical staining for TGF-ß1, α-smooth muscle actin, and collagen type I and type III demonstrated marked decrease compared to the animals without hADSC treatment. CONCLUSIONS: Treatment with hADSCs improved liver functions, markedly reduced hepatic fibrosis and early cirrhosis. Various pleiotropic and paracrine factors secreted from the hADSCs seem to serve as reparative functions in the attenuation of liver cirrhosis. The data demonstrated that treatment with hADSCs can be successfully used as a potent therapeutic method to prevent progression of hepatic fibrosis and related adverse events.

Inhibitory effect of taurine on rotator cuff degeneration via mitochondrial protection.

OBJECTIVES: Degenerative rotator cuff tears do not heal spontaneously, necessitating surgical intervention. This makes prevention crucial, but effective prophylactic measures are currently lacking. Oxidative stress has recently been implicated as a cause of degenerative rotator cuff tears, while mitochondrial injury has been reported in the development of age-related rotator cuff degeneration. Taurine, which has antioxidant properties, has been found to be effective in the treatment of various mitochondrial abnormalities. This prompted us to investigate the inhibitory effect of taurine and some other antioxidants against rotator cuff degeneration using tenocytes. METHODS: Hydrogen peroxide (H2O2, 2 mM) was added to tenocytes in medium with 0.8 µM taurine (Group TAU), medium with 100 µM α-tocopherol (Group E), and medium with 150 µM ascorbic acid (Group C), then each medium was cultured for 24 h. Tenocytes supplemented with 2 mM H2O2 alone were similarly cultured for 24 h (Group H2O2). In each group, immunostaining was performed for the oxidative stress marker 8-hydroxy-2'-deoxyguanosine and advanced glycation end products (AGE), which contribute to the development of age-related rotator cuff degeneration. In addition, levels of reactive oxygen species were measured using a cell-based assay kit, and results were compared. Immunostaining was also performed for indices of apoptosis (caspase-9, cleaved caspase-3 and Bcl-2), and Western blotting was used to quantify activation of caspase-9 at an early stage in each group. RESULTS: Oxidative stress and AGE levels were decreased in the E and C groups. Levels of all parameters were reduced in the TAU group. CONCLUSIONS: Taurine showed preventative effects against rotator cuff degeneration. The simple method of administration and paucity of side effects make clinical application easy, and the clear potential as a novel prophylactic strategy against degenerative rotator cuff tear warrants further study.

Solitary extrahepatic hepatocellular carcinoma in vertebrae without a primary lesion in the liver might originate from bone marrow: a case report and new hypothesis based on a review of the literature and the latest findings.

Solitary extrahepatic hepatocellular carcinoma without a primary lesion in the liver is rare and unique. In such patients, in addition to hepatocellular carcinoma, hepatoid adenocarcinoma, hepatoid teratoma, and hepatoid yolk sac tumor must be considered as differential diagnoses, and patients must be investigated in detail by histopathological studies with immunohistochemistry, especially using epithelial markers for which tumor cells are generally negative in hepatocellular carcinoma. A case with a solitary neoplasm of the vertebrae, which was diagnosed histopathologically as hepatocellular carcinoma, without a primary lesion is presented. The primary lesion was not identified even on autopsy, and the liver was pathologically almost normal. Given the review of the literature and circumstantial evidence, we would like to propose a bold new hypothesis that hepatocellular carcinoma might primarily originate from bone marrow.

Complete remission after concurrent single-agent S-1 chemoradiotherapy for stage IIIB esophageal cancer: an autopsy case report.

Chemoradiotherapy regimens for patients with esophageal cancer intolerant to standard therapies remain to be established. The standard therapy for patients with stage II-III esophageal squamous cell carcinoma, who are not surgical candidates, is definitive chemoradiotherapy with concomitant use of 5-fluorouracil and cisplatin; however, cisplatin can cause serious adverse events. An 83-year-old Japanese man developed a 2-month history of nausea and vomiting. Contrast-enhanced computed tomography revealed concentric wall thickening in the mid-to-lower esophagus with surrounding regional lymph node swelling. Upper gastrointestinal endoscopy revealed an ulcerated tumor with raised margins in the middle esophagus. He was diagnosed with stage IIIB (T3N2M0) esophageal squamous cell carcinoma, pathologically exhibiting squamous epithelium-like invasive abnormal structure with atypical cells. He underwent chemoradiotherapy involving four-dimensional conformal radiotherapy and single-agent S-1 rather than the standard chemoradiotherapy, and achieved clinical remission 2 months later on endoscopy and computed tomography. The patient died 1 year later due to pneumonia, and the autopsy did not reveal any evidence of squamous cell carcinoma in the esophagus, surrounding lymph nodes, or other organs, suggesting pathologically complete remission. Concurrent single-agent S-1 chemoradiotherapy may induce complete remission of stage IIIB esophageal cancer and is a possible alternative for older patients or those with multiple comorbidities.

Prognostic Impact of Sphingosine Kinase 1 in Nonsmall Cell Lung Cancer.

Bioactive sphingolipid is clearly relevant to lung physiology. The relationship of the bioactive sphingolipid pathway to pulmonary disease has been studied in cellular, tissue, and animal model, including lung cancer models. The samples of 53 patients diagnosed with nonsmall cell lung carcinoma (NSCLC) between June 2009 and May 2014 at our hospital were analyzed. Immunohistochemical (IHC) analysis was performed. The degree of immunostaining was reviewed and scored. Using this method of assessment, we evaluated the IHC score of sphingosine kinase 1 (SPHK1), vimentin, E-cadherin, and Ki-67. Both invasive adenocarcinoma cell and squamous cell carcinoma cell were well stained by SPHK1, and fibroblasts were also well stained by SPHK1. Although the IHC score of SPHK1 was not significantly differed between invasive adenocarcinoma and squamous cell carcinoma, the IHC scores of fibroblast, vimentin, and Ki-67 were higher in squamous cell carcinoma than invasive adenocarcinoma. Correlation among IHC scores in each of invasive adenocarcinoma and squamous cell carcinoma was performed. SPHK1 had positive correlation with both fibroblast and Ki-67, and fibroblast and Ki-67 had also positive correlation in invasive adenocarcinoma. On the contrary, SPHK1 had no significant correlation with fibroblast, and had negative correlation with Ki-67 in squamous cell carcinoma. Although there was not significant prognostic difference in SPHK1 score (P = .09), IHC score high group tended to be worse on relapse-free survival. SPHK1 might be prognostic factor in lung-invasive adenocarcinoma and novel target for drug against lung-invasive adenocarcinoma.

Resistance of bone marrow mesenchymal stem cells in a stressed environment - Comparison with osteocyte cells.

Introduction: Recently, the efficacy of mesenchymal stem cells (MSCs) mediated by their tissue repair and anti-inflammatory actions in the prevention and therapy of various disorders has been reported. In this research, our attention was focused specifically on the prevention and therapy of glucocorticoid-induced osteonecrosis. We investigated the stress resistance of MSC against glucocorticoid administration and hypoxic stress, which are factors known to induce osteocytic cell death. Materials and Methods: Mouse bone cells (MLO-Y4) and bone-marrow derived mouse MSCs were exposed to dexamethasone (Dex), hypoxia of 1% oxygen or both in vitro. Mitochondrial membrane potentials were estimated by mitochondria labeling with a cell-permeant probe (Mito tracker red); expression of these apoptosis-inducing molecules, oxidative stress marker (8-hydroxy-2'-deoxyguanosine), caspase-3, -9, and two apoptosis-inhibiting molecules, energy-producing ATP synthase (ATP5A) and X-linked inhibitor of apoptosis protein (XIAP), were analyzed by both immunofluorescence and western blot. Results: With exposure to either dexamethasone or hypoxia, MLO-Y4 showed reduced mitochondrial membrane potential, ATP5A and upregulation of 8-OHdG, cleaved caspases and XIAP. Those changes were significantly enhanced by treatment with dexamethasone plus hypoxia. In MSCs, however, mitochondrial membrane potentials were preserved, while no significant changes in the pro-apoptosis or anti-apoptosis molecules analyzed were found even with exposure to both dexamethasone and hypoxia. No such effects induced by treatment with dexamethasone, hypoxia, or both were demonstrated in MSCs at all. Discussion: In osteocyte cells subjected to the double stresses of glucocorticoid administration and a hypoxic environment osteocytic cell death was mediated via mitochondria. In contrast, MSC subjected to the same stressors showed preservation of mitochondrial function and reduced oxidative stress. Accordingly, even under conditions sufficiently stressful to cause the osteocytic cell death in vivo, it was thought that the function of MSC could be preserved, suggesting that in the case of osteonecrosis preventative and therapeutic strategies incorporating their intraosseous implantation may be promising.

Diffusion-weighted whole-body imaging with background suppression (DWIBS) is effective and economical for detection of metastasis or recurrence of lung cancer.

BACKGROUND: Diffusion-weighted whole-body imaging with background suppression (DWIBS) is used for the diagnosis and staging of cancers. The medical cost of an MR examination including DWIBS is $123, which is 80% less expensive than the cost ($798) of F18-fluorodeoxyglucose positron emission tomography/computed tomography (FDG-PET/CT) examination. METHODS: This study examined the efficacy of DWIBS for relapses after lung cancer resection. A total of 55 patients who had pulmonary resection of lung cancer, postoperative computed tomography (CT) every six months, and DWIBS and FDG-PET/CT (every year) were enrolled in this study. If a metastatic lesion was detected on CT scan, DWIBS and FDG-PET/CT were also used. RESULTS: A total of 55 patients who underwent pulmonary resections for lung cancer, and had CT, DWIBS and FDG-PET/CT examination during follow-up after pulmonary resection were enrolled in this study. Lung cancer in 32 patients relapsed. Postoperative radiographic examinations revealed pulmonary metastases in 17 patients, bone metastases in seven, liver metastases in five, lymph node metastases in five, pleural metastases in four, metastases to the chest wall in two, brain metastases in two, adrenal gland metastasis in one, and renal metastasis in one. The mean apparent diffusion coefficient (ADC) value of the relapse was 0.9 to 1.70 × 10-3 mm2 /s. The accuracy 0.98 (54/55) of DWIBS for detecting multiple metastatic lesions was likely to be higher than 0.94 (52/55) of CT or 0.94 (52/55) of FDG-PET/CT, but there were no significant differences. CONCLUSIONS: DWIBS can detect multiple metastatic lesions throughout the entire body and differentiate malignancy from benignity in only one examination. DWIBS has benefits of diagnostic accuracy and is less expensive in medical costs for the detection of a relapse. DWIBS could potentially replace FDG-PET/CT after lung cancer resection.

Differentiation between suture recurrence and suture granuloma after pulmonary resection for lung cancer by diffusion-weighted magnetic resonance imaging or FDG-PET / CT.

There has been no publication which supports the usefulness of DWI differentiating for suture recurrence and suture granuloma after resection for lung cancer. We presented efficacy of DWI or FDG-PET/CT for an assessment of suture lesions after resection for lung cancer. Thirteen suture recurrences and 15 suture granulomas were examined. There were 24 adenocarcinomas and 4 squamous cell carcinomas, and 26 partial resections and 2 segmentectomies. The period of time (907±907 days) between surgery and suture recurrence was not significantly longer than that (546±547 days) between surgery and suture granuloma. Diffusion detectability scores (a 5-point scale) of suture recurrences was significantly higher than that of suture granulomas. The ADC value (1.35±0.24 × 10-3mm2/sec) of suture recurrences was significantly lower than that (1.85±0.60 × 10-3mm2/sec) of suture granulomas. The SUVmax (6.1 ±â€¯5.0) of suture recurrences was not significantly higher than that (4.2 ±â€¯2.5) of suture granulmas. The sensitivity of 85% (11/13) with DWI was not significantly higher than 69% (9/13) with FDG-PET/CT for suture recurrences. The specificity of 73% (11/15) with DWI was not significantly higher than the 60% (9/15) with FDG-PET/CT for suture granulomas. The accuracy of 79% (22/28) with DWI was not significantly higher than that of 64% (18/28) with FDG-PET/CT for suture recurrences and granulomas. DWI can differentiate suture granuloma from suture recurrence after resection of lung cancer. DWI is more useful than FDG-PET/CT for the differentiation between suture recurrence and suture granuloma after resection for lung cancer.

Taurine Inhibits Glucocorticoid-Induced Bone Mitochondrial Injury, Preventing Osteonecrosis in Rabbits and Cultured Osteocytes.

Mitochondrial injury has recently been implicated in the pathogenesis of glucocorticoid-induced osteonecrosis. Using cultured osteocytes and a rabbit model, we investigated the possibility that taurine (TAU), which is known to play a role in the preservation of mitochondrial function, might also prevent the development of osteonecrosis. To reduplicate the intraosseous environment seen in glucocorticoid-induced osteonecrosis, dexamethasone (Dex) was added to MLO-Y4 cultured in 1% hypoxia (H-D stress environment). An in vitro study was conducted in which changes in mitochondrial transcription factor A (TFAM), a marker of mitochondrial function, and ATP5A produced by mitochondria, induced by the presence/absence of taurine addition were measured. To confirm the effect of taurine in vivo, 15 Japanese White rabbits were administered methylprednisolone (MP) 20 mg/kg as a single injection into the gluteus muscle (MP+/TAU- group), while for 5 consecutive days from the day of MP administration, taurine 100 mg/kg was administered to 15 animals (MP+/TAU+ group). As a control 15 untreated rabbits were also studied. The rabbits in each of the groups were sacrificed on the 14th day after glucocorticoid administration, and the bilateral femora were harvested. Histopathologically, the incidence of osteonecrosis was quantified immunohistochemically by quantifying TFAM and ATP5A expression. In the rabbits exposed to an H-D stress environment and in MP+/TAU- group, TFAM and ATP5A expression markedly decreased. With addition of taurine in the in vitro and in vivo studies, the expression of TFAM and ATP5A was somewhat decreased as compared with Dex-/hypoxia- or MP-/TAU- group, while improvement was noted as compared with Dex+/hypoxia+ or MP+/TAU- group. In rabbits, the incidence of osteonecrosis was 80% in MP+/TAU- group, in contrast to 20% in the taurine administered group (MP+/TAU+), representing a significant decrease. Since taurine was documented to exert a protective effect on mitochondrial function by inhibiting the mitochondrial dysfunction associated with glucocorticoid administration, we speculated that it might also indirectly help to prevent the development of osteonecrosis in this context. Since taurine is already being used clinically, we considered that its clinical application would also likely be smooth.

Inhibition of γ-glutamyltransferase ameliorates ischaemia-reoxygenation tissue damage in rats with hepatic steatosis.

BACKGROUND AND PURPOSE: Hepatic steatosis may be associated with an increased γ-glutamyltransferase (γ-GT) levels. Ischaemia-reoxygenation (IR) injury causes several deleterious effects. We evaluated the protective effects of a selective inhibitor of γ-GT in experimentally induced IR injury in rats with obesity and steatosis. EXPERIMENTAL APPROACH: Otsuka Long-Evans Tokushima Fatty (OLETF) rats with hepatic steatosis were used in the current study. The portal vein and hepatic artery of left lateral and median lobes were clamped to induce ischaemia. Before clamping, 1 ml of saline (IR group) or 1-ml saline containing 1 mg·kg-1 body weight of GGsTop (γ-GT inhibitor; IR-GGsTop group) was injected into the liver via the inferior vena cava. Blood flow was restored after at 30 min of the start of ischaemia. Blood was collected before, at 30 min after ischaemia and at 2 h and 6 h after reoxygenation. All the animals were killed at 6 h and the livers were collected. KEY RESULTS: Treatment with GGsTop resulted in significant reduction of serum ALT, AST and γ-GT levels and hepatic γ-GT, malondialdehyde, 4-hydroxy-2-nonenal and HMGB1 at 6 h after reoxygenation. Inhibition of γ-GT retained normal hepatic glutathione levels. There was prominent hepatic necrosis in IR group, which is significantly reduced in IR-GGsTop group. CONCLUSION AND IMPLICATIONS: Treatment with GGsTop significantly increased hepatic glutathione content, reduced hepatic MDA, 4-HNE and HMGB1 levels and, remarkably, ameliorated hepatic necrosis after ischaemia-reoxygenation. The results indicated that GGsTop could be an appropriate therapeutic agent to reduce IR-induced liver injury in obesity and steatosis.

Emergence of lung cancer with a low PD-L1 expression level after the administration of immune check point inhibitor for lung adenocarcinoma with a high PD-L1 expression level: A case report.

BACKGROUND: Checkpoint therapy against PD-1 has proven effective and positive results have been observed in several types of cancer, including lung cancer, renal cancer, lymphoma and melanoma. However, the effects of long-term ICI treatment remain insufficient and the development of resistance is an issue that remains to be solved. CASE PRESENTATION: A 70-year-old man was diagnosed with lung adenocarcinoma (stage IVB, T4N3M1c) with a high programmed death ligand-1 (PDL1) expression level (tumor proportion score [TPS]: 80% score at the time of the diagnosis, before treatment). At 16.5 months after the start of pembrolizumab, following the administration of 22 cycles of pembrolizumab, a new lesion appeared. Biopsy by video-assisted thoracic surgery (VATS) was performed for this lesion and a pathological diagnosis of lung adenocarcinoma with a low PD-L1 expression level. After the operation, pembrolizumab treatment was continued. The patient currently remains alive without disease progression at 20 months after the initial therapy. CONCLUSIONS: Our case highlights the importance of biopsy by VATS during immune checkpoint inhibitor (ICI) treatment when deciding the treatment strategy for newly confirmed tumors.

Mitochondrial Transcription Factor A added to Osteocytes in a Stressed Environment has a Cytoprotective Effect.

The main precipitant of glucocorticoid-associated femoral head osteonecrosis is widely accepted to be an ischemic-hypoxic event, with oxidative stress also as an underlying factor. Mitochondrial DNA is more vulnerable to oxidative injury than the nucleus, and mitochondrial transcription factor A (TFAM), which plays roles in its function, preservation, and regulation is being increasingly investigated. In the present study we focused on the impact of TFAM on the relation between the oxidative injury induced by the addition of glucocorticoid to a hypoxic environment and osteocytic cell necrosis. Using cultured osteocytes MLO-Y4 in a 1% hypoxic environment (hypoxia) to which 1µM dexamethasone (Dex) was added (Dex(+)/hypoxia(+)), an immunocytochemical study was conducted using 8-hydroxy-2'-deoxyguanosine (8-OHdG), an index of oxidative stress, and hypoxia inducible factor-1α (HIF-1α), a marker of hypoxia. Next, after adding TFAM siRNA, TFAM knockdown, cultured for 24h, and mitochondrial membrane potential were measured, they were stained with ATP5A which labels adenosine triphosphate (ATP) production. Dex was added to MLO-Y4 to which TFAM had been added, and cultured for 24h in hypoxia. The ratio of dead cells to viable cells was determined and compared. Enhanced expression of 8-OHdG, HIF-1α was found in osteocytes following the addition of glucocorticoid in a hypoxic environment. With TFAM knockdown, as compared to normoxia, mitochondrial function significantly decreased. On the other hand, by adding TFAM, the incidence of osteocytic cell necrosis was significantly decreased as compared with Dex(+)/hypoxia(+). TFAM was confirmed to be important in mitochondrial function and preservation, inhibition of oxidative injury and maintenance of ATP production. Moreover, prevention of mitochondrial injury can best be achieved by decreasing the development of osteocytic cell necrosis.