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Human platelets inhibit liver fibrosis in severe combined immunodeficiency mice

World Journal of Gastroenterology 2013年第32期19卷 5250-5260页

作者： Kazuhiro Takahashi Soichiro Murata kiyoshi fukunaga Nobuhiro Ohkohchi Department of Surgery University of Tsukuba

AIM:To investigate the role of human platelets in liver fibrosis.METHODS:Severe combined immunodeficiency(SCID)mice were administered CCl4and either phosphate-buffered saline(PBS group)or human platelet transfusions(hPLT group).Concentrations of hepatocyte growth factor(HGF),matrix metallopeptidases(MMP)-9,and transforming growth factor-β(TGF-β)in the liver tissue were compared between the PBS and the hPLT groups by enzyme-linked immunosorbent assay(ELISA)and Western blotting.The effects of a human platelet transfusion on liver fibrosis included the fibrotic area,hydroxyproline content,and-smooth muscle actin(α-SMA)expression,which were evaluated by picrosirius red staining,ELISA,and immunohistochemical staining using an anti-mouse-SMA antibody,respectively.Phosphorylations of mesenchymal-epithelial transition factor(Met)and SMAD3,downstream signals of HGF and TGF-β,were compared between the two groups by Western blotting and were quantified using densitometry.Hepatocyte apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling.Furthermore,the accumulation of human platelets in the liver 2 h after platelet transfusion was compared between normal and fibrotic livers by immunohistochemical staining using an anti-human CD41 antibody.RESULTS:The fibrotic area and hydroxyproline content in the liver were both significantly lower in the hPLT group when compared to the PBS group(fibrotic area,1.7%±0.6%vs 2.5%±0.6%,P=0.03;hydroxyproline content,121±26 ng/g liver vs 156±47 ng/g liver,P=0.04).There was less α-smooth muscle actin staining in the hPLT group than in the PBS group(0.5%±0.1%vs 0.8%±0.3%,P=0.02).Hepatic expression levels of mouse HGF and MMP-9were significantly higher in the hPLT group than in the PBS group(HGF,109±13 ng/g liver vs 88±22 ng/g liver,P=0.03;MMP-9,113%±7%/GAPDH vs 92%±11%/GAPDH,P=0.04).In contrast,the concentration of mouse TGF-β in the liver tissue was significantly lower in the hPLT group than in the PBS group(22±5ng/g liver vs 39±6 ng/g liver,P=0.02).Phosphorylation

关键词： Human platelet Liver fibrosis Hepatocyte apoptosis Hepatocyte growth factor Transforming growth factor-β Matrix metallopeptidases

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Three-dimensional imaging identified the accessory bile duct in a patient with cholangiocarcinoma

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World Journal of Gastroenterology 2014年第32期20卷 11451-11455页

作者： Ryoichi Miyamoto Yukio Oshiro Shinji Hashimoto Keisuke Kohno kiyoshi fukunaga Tatsuya Oda Nobuhiro Ohkohchi Division of Gastroenterological and Hepatobiliary Surgery and Organ Transplantation Department of Surgery University of Tsukuba Ibaraki 305-8575 Japan

The development of diagnostic imaging technology, such as multidetector computed tomography(MDCT) and magnetic resonance cholangiopancreatography(MRCP), has made it possible to obtain detailed images of the bile duct. Recent reports have indicated that a 3-dimensional(3D) reconstructed imaging system would be useful for understanding the liver anatomy before surgery. We have investigated a novel method that fuses MDCT and MRCP images. This novel system easily made it possible to detect the anatomical relationship between the vessels and bile duct in the portal hepatis. In this report, we describe a very rare case of extrahepatic cholangiocarcinoma associated with an accessory bile duct from the caudate lobe connecting with the intrapancreatic bile duct. We were unable to preoperatively detect this accessory bile duct using MDCT and MRCP. However, prior to the second operation, we were able to clearly visualise the injured accessory bile duct using our novel 3D imaging modality. In thisreport, we suggest that this imaging technique can be considered a novel and useful modality for understanding the anatomy of the portal hepatis, including the hilar bile duct.

关键词： 3-dimensional imaging Hepatobiliary and pancreatic

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Influence of Kupffer cells and platelets on ischemia-reperfusion injury in mild steatotic liver

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World Journal of Gastroenterology 2013年第9期19卷 1396-1404页

作者： Koichi Ogawa Tadashi Kondo Takafumi Tamura Hideki Matsumura kiyoshi fukunaga Tatsuya Oda Nobuhiro Ohkohchi Department of Surgery Graduate School of Comprehensive Human Sciences University of Tsukuba Tsukuba 305-8575 Japan

AIM: To investigate the effect of mild steatotic liver on ischemia-reperfusion injury by focusing on Kupffer cells (KCs) and platelets. METHODS: Wistar rats were divided into a normal liver group (N group) and a mild steatotic liver group (S group) induced by feeding a choline-deficient diet for 2 wk. Both groups were subjected to 20 min of warm ischemia followed by 120 min of reperfusion. The number of labeled KCs and platelets in sinusoids and the blood perfusion in sinusoids were observed by intravital microscopy (IVM), which was performed at 30, 60 and 120 min after reperfusion. To evaluate serum alanine aminotransferase as a marker of liver deterioration, blood samples were taken at the same time as IVM.RESULTS: In the S group, the number of platelets adhering to KCs decreased significantly compared with the N group (120 after reperfusion; 2.9±1.1 cells/acinus vs 4.8±1.2 cells/acinus, P<0.01). The number of KCs in sinusoids was significantly less in the S group than in the N group throughout the observation periods (before ischemia, 19.6±3.3 cells/acinus vs 28.2±4.1 cells/acinus, P<0.01 and 120 min after reperfusion, 29.0±4.3 cells/acinus vs 40.2±3.3 cells/acinus, P<0.01). The blood perfusion of sinusoids 120 min after reperfusion was maintained in the S group more than in the N group. Furthermore, elevation of serum alanine aminotransferase was lower in the S group than in the N group 120 min after reperfusion (99.7±19.8 IU/L vs 166.3±61.1 IU/L, P=0.041), and histological impairment of hepatocyte structure was prevented in the S group. CONCLUSION: Ischemia-reperfusion injury in mild steatotic liver was attenuated compared with normal liver due to the decreased number of KCs and the reduction of the KC-platelet interaction.

关键词： Steatotic liver Mild steatotic liver Kupffer cell Platelet Ischemia-reperfusion Intravital microscopy

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Novel 3-dimensional virtual hepatectomy simulation combined with real-time deformation

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World Journal of Gastroenterology 2015年第34期21卷 9982-9992页

作者： Yukio Oshiro Hiroaki Yano Jun Mitani Sangtae Kim Jaejeong Kim kiyoshi fukunaga Nobuhiro Ohkohchi Department of Surgery Division of Gastroenterological and Hepatobiliary Surgery and Organ TransplantationFaculty of MedicineUniversity of Tsukuba Department of Computer Science Graduate School of System Information EngineeringUniversity of Tsukuba School of Informatics College of Media Arts Science and TechnologyFaculty of Library Information and Media ScienceUniversity of Tsukuba

AIM:To develop a novel 3-dimensional(3D) virtual hepatectomy simulation software,Liversim,to visualize the real-time deformation of the liver.METHODS:We developed a novel real-time virtual hepatectomy simulation software program called Liversim. The software provides 4 basic functions:viewing 3D models from arbitrary directions,changing the colors and opacities of the models,deforming the models based on user interaction,and incising the liver parenchyma and intrahepatic vessels based on user operations. From April 2010 through 2013,99 patients underwent virtual hepatectomies that used the conventional software program SYNAPSE VINCENT preoperatively. Between April 2012 and October 2013,11 patients received virtual hepatectomies using the novel software program Liversim; these hepatectomies were performed both preoperatively and at the same that the actual hepatectomy was performed in an operating room. The perioperative outcomes were analyzed between the patients for whom SYNAPSE VINCENT was used and those for whom Liversim wasused. Furthermore,medical students and surgical residents were asked to complete questionnaires regarding the new software.RESULTS:There were no obvious discrepancies(i.e.,the emergence of branches in the portal vein or hepatic vein or the depth and direction of the resection line) between our simulation and the actual surgery during the resection process. The median operating time was 304 min(range,110 to 846) in the VINCENT group and 397 min(range,232 to 497) in the Liversim group(P = 0.30). The median amount of intraoperative bleeding was 510 m L(range,18 to 5120) in the VINCENT group and 470 m L(range,130 to 1600) in the Liversim group(P = 0.44). The median postoperative stay was 12 d(range,6 to 100) in the VINCENT group and 13 d(range,9 to 21) in the Liversim group(P = 0.36). There were no significant differences in the preoperative outcomes between the two groups. Liversim was not found to be clinically inferior to SYNAPSE VINCENT. Both students and surgical residents reported that th

关键词： Liver Surgery Simulation Virtual hepatectomy Real-

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New drug delivery system for liver sinusoidal endothelial cells for ischemia-reperfusion injury

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World Journal of Gastroenterology 2015年第45期21卷 12778-12786页

作者： Naoki Sano Takafumi Tamura Naoyuki Toriyabe Takeshi Nowatari Ken Nakayama Tomohito Tanoi Soichiro Murata Yu Sakurai Mamoru Hyodo kiyoshi fukunaga Hideyoshi Harashima Nobuhiro Ohkohchi Department of Surgery Doctoral Program in Clinical Science Graduate School of Comprehensive Human Sciences University of Tsukuba Faculty of Pharmaceutical Sciences Hokkaido University

AIM: To investigate the cytoprotective effects in hepatic ischemia-reperfusion injury, we developed a new formulation of hyaluronic acid(HA) and sphingosine 1-phophate.METHODS: We divided Sprague-Dawley rats into 4 groups: control, HA, sphingosine 1-phosphate(S1P), and HA-S1 P. After the administration of each agent, we subjected the rat livers to total ischemia followed by reperfusion. After reperfusion, we performed the following investigations: alanine aminotransferase(ALT), histological findings, Td T-mediated d UTP-biotin nick end labeling(TUNEL) staining, and transmission electron microscopy(TEM). We also investigated the expressionof proteins associated with apoptosis, hepatoprotection, and S1 P accumulation. RESULTS: S1 P accumulated in the HA-S1 P group livers more than S1 P group livers. Serum ALT levels, TUNEL-positive hepatocytes, and expression of cleaved caspase-3 expression, were significantly decreased in the HA-S1 P group. TEM revealed that the liver sinusoidal endothelial cell(LSEC) lining was preserved in the HA-S1 P group. Moreover, the HA-S1 P group showed a greater increase in the HO-1 protein levels compared to the S1 P group.CONCLUSION: Our results suggest that HA-S1 P exhibits cytoprotective effects in the liver through the inhibition of LSEC apoptosis. HA-S1 P is an effective agent for hepatic ischemia/reperfusion injury.

关键词： Hyaluronic acid Sphingosine 1-phosphate Liver sinu

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Useful detection of CD147 (EMMPRIN) for pathological diagnosis of early hepatocellular carcinoma in needle biopsy samples

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World Journal of Gastroenterology 2007年第21期13卷 2913-2917页

作者： Satoshi Mamori Keisuke Nagatsuma Tomokazu Matsuura kiyoshi Ohkawa Hiroshi Hano Masaharu fukunaga Masato Matsushima Yoshifumi Masui Nao Fushiya Hiroshi Onoda Yasuyuki Searashi Ichiro Takagi Hisao Tajiri Division of Gastroenterology and Hepatology Department of Internal MedicineThe Jikei University School of MedicineTokyo 105-8461Japan Department of Laboratory Medicine The Jikei University School of MedicineTokyo 105-8461Japan Department of Biochemistry 1 The Jikei University School of MedicineTokyo 105-8461Japan Department of Pathology The Jikei University School of MedicineTokyo 105-8461Japan Department of Pathology The Jikei University School of MedicineThe Daisan HospitalTokyo 105-8461Japan Division of Clinical Research and development The Jikei University School of MedicineTokyo 105-8461Japan

AIM:To make clear whether CD147 (EMMPRIN) expression in pathological tumor samples with a fine-needle aspiration biopsy is useful for pathological diagnosis of early hepatocellular carcinoma (HCC). METHODS:Twenty-two patients (15 men and 7 women; median age 68 years,range 56-81 years) underwent a liver tissue biopsy in order to make a diagnosis of HCC. Paraffin-embedded liver biopsy tissue samples from 22 patients were stained with anti-CD147 antibody,murine monoclonal antibody 12C3 (MAb12C3) for immunohistochemical analysis. An immunohistochemical analysis of CD147 was performed and the degree of staining compared between tumor and non-tumor tissue. In addition,the degree of staining within tumor tissue was compared according to a number of clinicopathological variables. RESULTS:The degree of staining of CD147 was significantly higher in tumor tissues than non-tumor tissues,even in tumors less than 15 mm in diameter.The expression of this protein was significantly elevated in HCC tissue specimens from patients with a low value of serum AST and γ-GTP.

关键词： CD147 肝癌早期诊断病理组织活检

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