Hidetake Amemiya

Role of IL-17A in Neutrophil Recruitment and Hepatic Injury after Warm Ischemia–Reperfusion Mice

Recent evidence suggests that IL-17A regulates neutrophil-dependent organ injury. Accordingly, the purpose of this study was to determine the role of IL-17A in neutrophil recruitment after ischemia–reperfusion (I/R) and in subsequent liver injury. Two mouse models including wild-type and IL-17A knockout mice were evaluated for I/R injury. The medial largest lobe of the liver was clamped for 90 min. In another set of experiments, recombinant mouse (rm)IL-17A homodimer or rmIL-17A/F heterodimer were administered to knockout mice before I/R, and liver injury was investigated. Isolated Kupffer cells were incubated with rmIL-17A or rmIL-17F, and production of TNF-α was measured. Studies evaluating the extent of liver injury as measured by serum transaminase levels demonstrated similar levels in the acute phase (6 h) in these two models. In contrast, in the subacute phase (20 h) after I/R, both serum transaminase levels and percent of hepatic necrosis were significantly reduced in the knockout mice compared with the wild-type mice. This reduction in liver injury seen in the knockout mice was associated with suppression of chemokine and adhesion molecule expression and reduction in infiltration of neutrophils into the liver. Administration of rmIL-17A homodimer, but not IL-17A/F heterodimer, increased liver injury in the subacute phase of I/R in KO mice. TNF-α production by isolated Kupffer cells increased significantly in the cells incubated with rmIL-17A compared with rmIL-17F. These results indicate that IL-17A is a key regulator in initiating neutrophil-induced inflammatory responses and hepatic injury in the subacute phase after reperfusion.

Role of interleukin-18 and its receptor in hepatocellular carcinoma associated with hepatitis C virus infection

Interleukin (IL)‐18 is a proinflammatory cytokine that is up‐regulated in patients with hepatitis C virus (HCV) infection, which is the most common underlying disease in hepatocellular carcinoma (HCC). The purpose of our study was to investigate the role of IL‐18 in HCC associated with HCV infection. Sixty‐five patients with HCC and HCV infections who received curative surgical resections were examined in our study. The expression of the IL‐18 receptor was investigated in HCC tissues obtained from these patients and in 2 HCC cell lines. Nuclear factor (NF)‐κB activity and the expression of Bcl‐xL and xIAP mRNA were tested in the cell lines using recombinant human (rh) IL‐18. The IL‐18 receptor was expressed in both the HCC tissues and the cell lines. NF‐κB activation and the expression of Bcl‐xL and xIAP mRNA were increased by rhIL‐18. Moreover, rhIL‐18 suppressed the apoptosis of HCC cells which was induced by etoposide in vitro. The overall survival rate (55.4%) was significantly worse in the IL‐18 receptor‐positive patients than in the IL‐18 receptor‐negative patients (p = 0.015). In a Cox multivariate analysis, the expression of the IL‐18 receptor was found to be a significant predictor of a poor outcome in HCC patients. The expression of the IL‐18 receptor and an antiapoptotic mechanism involving NF‐κB activation in HCC cells may be implicated in a poor patient outcome.

Liver Regeneration is Impaired in Macrophage Colony Stimulating Factor Deficient Mice After Partial Hepatectomy: The Role of M-CSF-Induced Macrophages

Macrophage colony stimulating factor (M-CSF), which induces maturation of macrophages, is notably expressed in the liver. Thus, the specific purpose of this study was to investigate the role of M-CSF in liver regeneration after partial hepatectomy (PH). Osteopetrotic (op/op) mice, genetically lacking functional M-CSF, or their littermate mice underwent 70% PH. Animals were sacrificed at the designated time points after PH, and remnant liver tissues were harvested for further investigations. Proliferation of hepatocytes was evaluated by the expression of BrdU and the liver-body weight ratio. The mRNA expression levels of TNF-α and IL-6 and protein expression levels of phosphorylated (p) STAT3 were measured. The number of Kupffer cells (KCs) was determined by immunohistochemistry. Furthermore, KCs were isolated from op/op mice or littermate mice, and mRNA expression levels of TNF- α and IL-6 were assessed after stimulation with LPS. In littermate mice, steady liver regeneration was observed. The number of KCs reduced markedly by about 60% in the op/op mice compared with littermates as reported previously. Furthermore, these cells were morphologically small and immature. In littermate mice, the peak expression levels of TNF-α and IL-6 in the liver was observed 1h after PH, which was consistent with data in previous reports. In contrast, in op/op mice, the peak expression levels were observed 3 h after PH and were significantly lower compared with littermate mice. As a result, the proliferation of hepatocytes was significantly impaired in op/op mice. The mRNA expression level of IL-6, but not TNF-α,was significantly reduced in isolated KCs from op/op mice compared with the littermates after stimulation with LPS, suggesting that the function of KCs is different between op/op mice and littermate mice. To clarify the role of M-CSF in liver regeneration, op/op mice received intraperitoneally, mouse recombinant M-CSF 2 d before PH, and liver regeneration was also assessed. As a result, the numbers of Kupffer cells and liver regeneration were recovered in the op/op mice treated with M-CSF to a similar extent to those in their littermates. Thus, M-CSF-induced hepatic macrophages play an important role in liver regeneration after PH.

The Kupffer cell protects against acute lung injury in a rat peritonitis model: role of IL-10.

The possibility that Kupffer cells (KCs) play key beneficial and deleterious roles in multiple organ injury in sepsis has been discussed. The role of KCs in lung injury in a rat peritonitis model was investigated. Specifically, the involvement of interleukin (IL)‐10, which has anti‐inflammatory effects, was examined. Rats were given saline or gadolinium chloride (GdCl3), a KC toxicant, 24 h before cecal ligation and puncture (CLP). Survival was assessed for 7 days after CLP. The liver, lung, and serum were harvested, and the expression of cytokines was assessed. Macrophages were isolated from each organ after CLP, and the mRNA expression of inflammatory mediators was assessed. GdCl3 treatment increased lung injury and mortality. Plasma endotoxin levels were significantly greater, whereas serum IL‐10 levels were lower in the GdCl3 than in the control group after CLP. IL‐10 levels were significantly greater in the aorta than the hepatic vein. The mRNA expression of IL‐10 was less in KCs from the GdCl3 than the control group. In the liver, the expression of IL‐10 increased rapidly and continuously, up to 9 h in the control group, but values were significantly lower in the GdCl3 group. Rabbit anti‐rat IL‐10 antibodies were injected just after CLP to investigate the effects of immunoneutralization of endogenously produced IL‐10. In the antibody‐treated group, lung injury and mortality increased compared with animals treated with rabbit immunoglobulin G. Taken together, these results indicate that KCs play a protective role in lung injury in sepsis by production of IL‐10.

Lens culinaris agglutinin-reactive fraction of AFP is a useful prognostic biomarker for survival after repeat hepatic resection for HCC

Background and Aim:  Repeat hepatic resection for recurrent hepatocellular carcinoma (HCC) is effective in improving long‐term outcome in selected patients. In the present study, we attempted to identify the prognostic factors influencing overall and recurrence‐free survival after the second hepatic resection.

Preoperative Gadoxetic Acid-Enhanced MRI and Simultaneous Treatment of Early Hepatocellular Carcinoma Prolonged Recurrence-Free Survival of Progressed Hepatocellular Carcinoma Patients after Hepatic Resection

Background/Purpose. The purpose of this study was to clarify whether preoperative gadoxetic acid-enhanced magnetic resonance imaging (EOB-MRI) and simultaneous treatment of suspected early hepatocellular carcinoma (eHCC) at the time of resection for progressed HCC affected patient prognosis following hepatic resection. Methods. A total of 147 consecutive patients who underwent their first curative hepatic resection for progressed HCC were enrolled. Of these, 77 patients underwent EOB-MRI (EOB-MRI (+)) before hepatic resection and the remaining 70 patients did not (EOB-MRI (−)). Suspected eHCCs detected by preoperative imaging were resected or ablated at the time of resection for progressed HCC. Results. The number of patients who underwent treatment for eHCCs was significantly higher in the EOB-MRI (+) than in the EOB-MRI (−) (17 versus 6; P = 0.04). Recurrence-free survival (1-, 3-, and 5-year; 81.4, 62.6, 48.7% versus 82.1, 41.5, 25.5%, resp., P < 0.01), but not overall survival (1-, 3-, and 5-year; 98.7, 90.7, 80.8% versus 97.0, 86.3, 72.4%, resp., P = 0.38), was significantly better in the EOB-MRI (+). Univariate and multivariate analyses showed that preoperative EOB-MRI was one of the independent factors significantly correlated with better recurrence-free survival. Conclusions. Preoperative EOB-MRI and simultaneous treatment of eHCC prolonged recurrence-free survival after hepatic resection.

Non‐occlusive mesenteric ischemia after chemotherapy for metastatic melanoma

genetic test and performed a mutation analysis of the NF1 gene. Total RNA was extracted from the peripheral blood and constitutional DNA was synthesized using reverse transcription polymerase chain reaction (PCR). The PCR product was directly sequenced and the base sequence compared against records stored in GenBank (NC_000017.10, NM_000267.3) using GENETYX version 11 software (GENETYX, Tokyo, Japan). The deletion of c.7808_8053del was identified in exon 54 to exon 56, suggesting that this deletion caused truncated neurofibromin molecules due to an in-frame deletion. (Fig. 1b). Only six cases of non-segmental, late-onset NF1, including our patient, have been reported. The onset of the pigment spots and neurofibromas in all of the reported cases occurred in the third decade of life or later, and there was no family history. Most of these cases revealedmild symptomswithout Lisch nodules. Mosaicism may be responsible for the late onset of hereditary diseases. Mosaicism is defined as the presence of two or more populations of cells with different genotypes in one individual. An example of typical mosaicism is segmental NF1, which is caused by a postzygotic mutation of NF1. Later, somatic mutations may cause localized diseases such as the segmental form, while early somatic mutations may give rise to a generalized cutaneous form. A generalized form of mosaicism has been reported to present generalized but milder manifestations such as fewer neurofibromas and/or pigment spots. Other possible causes of mild manifestations may include the benign nature of a leaky NF1-splice mutation. A specific, small mutation, a 3-bp in-frame deletion in exon 17 of the NF1 gene, has been identified in a milder phenotype of NF1. Later onset of the cutaneous manifestations and the absence of ophthalmologic manifestations may arise from mosaicism or specific, small mutations, though we were unable to confirm the presence of them in our case.

Pancreaticoduodenectomy for pancreas carcinoma occurring in the annular pancreas: report of a case.

The annular pancreas is a rare congenital anomaly in which a ring of the pancreas parenchyma surrounds the second part of the duodenum. Malignant tumors are extremely rare in patients with an annular pancreas. A 64-year-old man presented with appetite loss and vomiting. Abdominal contrast-enhanced computed tomography (CT) indicated pancreas parenchyma surrounding the second part of the duodenum, and a hypovascular area occupying lesion in the annular pancreas. Subtotal stomach-preserving pancreaticoduodenectomy was performed. Histopathology showed pancreatic carcinoma occurring in the complete annular pancreas.

miR-122-5p as a novel biomarker for alpha-fetoprotein-producing gastric cancer

AIM To investigate the clinical utility of alpha-fetoprotein (AFP)-producing gastric cancer (AFPGC)-specific microRNA (miRNA) for monitoring and prognostic prediction of patients. METHODS We performed a comprehensive miRNA array-based approach to compare miRNA expression levels between AFP-positive and AFP-negative cells in three patients with primary AFPGC. We next examined the expression levels of the selected miRNAs in five AFPGC and ten non-AFPGC tissue samples by quantitative reverse transcription-polymerase chain reaction to validate their utility. We also investigated the expression levels of the selected miRNA not only in tissue but also in plasma samples. Moreover, we investigated the relationship between plasma AFP levels and plasma selected miRNA expression levels, and also investigated the correlation of the selected miRNA expression levels and malignant potential. RESULTS Among the five miRNAs selected from the miRNA array results, the expression levels of miR-122-5p were significantly higher in the AFPGC patients than in the non-AFPGC patients (P < 0.05). In tissue samples, miR-122-5p expression level tended to be lower in the non-AFPGC tissue than the normal gastric mucosa. Conversely, in the AFPGC tissue, miR-122-5p expression level was significantly higher in the AFPGC tissue than both the normal gastric mucosa and the non-AFPGC tissue samples (P < 0.05). Plasma miR-122-5p expression levels were also significantly higher in the AFPGC patients than the health volunteers and the non-AFPGC patients (P < 0.05) and were strongly correlated with plasma AFP levels (r = 0.7975, P < 0.0001). Moreover, the correlation of miR-122-5p expression in tissue samples with malignant potential was stronger than that of plasma AFP level in the AFPGC patients. In contrast, no correlation was found between miR-122-5p expression levels and liver metastasis in the non-AFPGC patients. CONCLUSION miR-122-5p might be a useful biomarker for early detection and disease monitoring in AFPGC.

Successful laparoscopic partial gastrectomy and spleen-preserving distal pancreatectomy for gastric duplication cyst connecting with the pancreatic tail

Highlights • Gastric duplication cyst(GDC) contiguous with the stomach and pancreatic tail is extremely rare.• GDCs are usually diagnosed at a younger age. In adults, they are very rare disease and the diagnosis may be difficult.• Surgical resection is considered to be the best treatment due to the difficulty of diagnosis.• Laparoscopic surgery is less invasive and should be selected whenever possible.