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Edaravone inhibits apoptosis caused by ischemia/reperfusion injury in a porcine hepatectomy model

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World Journal of Gastroenterology 2012年第27期18卷 3520-3526页

作者： Mitsugi Shimoda Yoshimi Iwasaki Toshie Okada keiichi kubota Second Department of Surgery Dokkyo Medical University880 Kita KobayashiMibuTochigi 321-0293Japan

AIM： TO investigate the effect of E3-methyl-l-phe- nyl-2-pyrazolin-5-one （Edr） on hepatic ischemia-reper- fusion （I/R） injury and liver regeneration in a porcine hepatectomy model. METHODS： One hour ischemia was induced by occlud- ing the vessels and the bile duct of the right and median lobes. A 40% left hepatectomy was performed after re- perfusion. Six animals received Edr （3 mg/kg per hour） intravenously and six control animals received saline just before reperfusion. Remnant liver volume, hemody- namics, aspartate aminotransferase （AST）, alanine ami- notransferase, lactate dehydrogenase and lactic acid, were compared between the groups. The expression of transforming growth factor-β （TGF-β1） and toll-like receptor （TRL） mRNA in hepatic tissues was examined using reverse transcription polymerase chain reaction. Apoptosis was demonstrated by terminal deoxynucleo- tidyl transferase dUTP nick end labeling （TUNEL） stain- ing, respectively. RESULTS： Serum AS-I- （P = 0.029）, and toll like recep- tor 4 level （P = 0.043） were significantly lower after 3 hin animals receiving Edr. In addition, TUNEL staining in Edr-treated pigs showed significantly fewer hepatocytes undergoing apoptosis compared with control pigs. After 1 mo, all factors were non-significantly different between the two groups. CONCLUSION： Edr is considered to reduce hepatic injury in the early stage of I/R injury in a porcine model.

关键词： Edaravone Ischemia-reperfusion injury Liver resection Transforming growth factor-13 Toll likereceptor 4

Preoperative portal vein embolization for hepatocellular carcinoma: consensus and controversy

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World Journal of Hepatology 2016年第9期8卷 439-445页

作者： Taku Aoki keiichi kubota Second Department of Surgery Dokkyo Medical University

Thirty years have passed since the first report of portal vein embolization(PVE),and this procedure is widely adopted as a preoperative treatment procedure for patients with a small future liver remnant(FLR).PVE has been shown to be useful in patients with hepatocellular carcinoma(HCC)and chronic liver ***,special caution is needed when PVE is applied prior to subsequent major hepatic resection in cases with cirrhotic livers,and volumetric analysis of the liver segments in addition to evaluation of the liver functional reserve before PVE is mandatory in such *** in the embolic material and selection of the treatment approach,and combined use of PVE and transcatheter arterial embolization/chemoembolization have yielded improved outcomes after PVE and major hepatic resections.A novel procedure termed the associating liver partition and portal vein ligation for staged hepatectomy has been gaining attention because of the rapid hypertrophy of the FLR observed in patients undergoing this procedure,however,application of this technique in HCC patients requires special caution,as it has been shown to be associated with a high morbidity and mortality even in cases with essentially healthy livers.

关键词： Hepatocellular carcinoma Future liver remnant Poral vein embolization Liver functional reserve The associating liver partition and portal vein ligation for staged hepatectomy

Erythropoietin ameliorates early ischemia-reperfusion injury following the Pringle maneuver

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World Journal of Gastroenterology 2010年第38期16卷 4838-4845页

作者： Masato Kato Tokihiko Sawada Junji Kita Mitsugi Shimoda keiichi kubota Second Department of Surgery Dokkyo Medical UniversityKitakobayash 880

AIM:To investigate the protective effect of erythropoietin (Epo) against ischemia-reperfusion injury (IR/I) following the Pringle maneuver (PM),in comparison with conventional steroid administration in a prospective randomized trial. METHODS:Patients were randomized by age, sex, diagnosis, and surgical method, and assigned to three groups:(1) A steroid group (STRD, n= 9) who received 100 mg of hydrocortisone before PM, and on postoperative days 1, 2 and 3, followed by tapering until postoperative day 7; (2) An EPO1 group (n=10) who received 30 000 U of Epo before the PM and at the end of surgery; and (3) An EPO2 group (n=8) who received 60 000 U of Epo before the PM. Hemoglobin (Hb), hematocrit (Ht), aspartate aminotransferase (AST), alanine transaminase (ALT),lactate dehydrogenase (LDH), lactate, interleukin-6 (IL-6),and tumor necrosis factor(TNF)-α were measured before and just after (Day 0) surgery, and on postoperative days 1, 3, 7 and 14. RESULTS: There were no increases in Hb and Ht in the EPO1 and EPO2 groups. AST was signif icantly lower in EPO1 than in STRD on Day 0 (P=0.041), and lower in EPO1 than in STRD and EPO2 on Day 1 (P=0.018). ALT was signif icantly lower in EPO1 than in STRD and EPO2 on Day 0 (P=0.020) and Day 1 (P=0.004). There were no signif icant inter-group differences in the levels of LDH and lactate. IL-6 was signif icantly lower in EPO1 than in STRD and EPO2 on Day 0 (P=0.0036) and Day 1 (P=0.0451). TNF-α was signif icantly lower in EPO1 than in STRD and EPO2 on Day 0 (P=0.0006) and Day 1 (P<0.0001). Furthermore, hospitalization was signif icantly shorter in EPO1 and EPO2 than in ***:Epo has greater potential than steroids to ameliorate IR/I after the PM. Epo at a dose of 30000 U, administered before PM and just after surgery, yields better results.

关键词： Erythropoietin Hepatic resection Pringle maneuver Steroid Prospective randomized study

Hepatocellular carcinoma with chronic B-type hepatitis complicated by autoimmune hemolytic anemia:A case report

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World Journal of Gastroenterology 2007年第32期13卷 4401-4404页

作者： Toshie Okada keiichi kubota Junji Kita Masato Kato Tokihiko Sawada Department of Gastro-enterological Surgery Dokkyo University School of MedicineTochigiJapan

A 57-year-old man consulted a local hospital because of a persistent slight fever. At the age of 37 years he was diagnosed having B-type hepatitis, but left the liver dysfunction untreated. Twenty years later, he was diagnosed having chronic hepatitis B, hepatocellular carcinoma （HCC） and macrocytic anemia, and referred to our hospital for further investigation. A HCC with a maximum diameter of 5.2 cm was detected in segment 8. Results of blood tests included 1.8 mg/dL serum total bilirubin, 0.9 mg/dL bilirubin, less than 10 mg/dL haptoglobin, 7.9 g/dL hemoglobin, 130 fL MCV, and 14.5% reticuloo/tes. A bone marrow sample showed erythroid hyperplasia. The direct Coombs test gave a positive result. We diagnosed the anemia as autoimmmune hemolytic anemia （AIHA）, for which prednisolone could not be administered due to positivity for HBsAg and HBeAg. After preparation of washed blood cells for later transfusion, the patient underwent systematic resection of segment 8. The cut surface of the resected specimen demonstrated an encapsulated yellow-brownish tumor measuring 52 mmx 40 mm which was diagnosed pathologicaly as moderately differentiated HCC. On the 9th postoperative day, the patient＇s temperature rose to 38℃, and exacerbated hemolysis was observed. The maximum total bilirubin value was 5.8 mg/dL and minimum hemoglobin level was 4.6 g/dL. He tolerated this period without blood transfusion. Currently he is being followed up as an outpatient, and shows no signs of HCC recurrence or symptoms of anemia. AIHA associated with HBV infection has been described in only three previous cases, and the present case is the first in which surgery was performed for accompanying HCC.

关键词： Hepatocellular carcinoma B-type hepatitis Auto-immune hemolytic anemia

MK615 inhibits pancreatic cancer cell growth by dual inhibition of Aurora A and B kinases

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World Journal of Gastroenterology 2008年第9期14卷 1378-1382页

作者： Toshie Okada Tokihiko Sawada Tatsushi Osawa Masakazu Adachi keiichi kubota Second Department of Surgery Dokkyo University School of Medicine Kitakobayashi 880MibuShimotsugaTochigi 321-0293Japan Second Department of Surgery Dokkyo University School of Medicine Kitakobayashi 880MibuShimotsuga Tochigi 321-0293Japan Japan Apricot Co.Ltd.1-11-7 MidorichoTakasakiGunma 370-0073 Japan Japan Apricot Co. Ltd.1-11-7 MidorichoTakasakiGunma 370-0073 Japan Second Department of Surgery Dokkyo University School of MedicineKitakobayashi 880Mibu ShimotsugaTochigi 321-0293Japan

AIM:To investigate the anti-neoplastic effect of MK615, an anti-neoplastic compound isolated from Japanese apricot, against human pancreatic cancer cells in vitro. METHODS: Three human pancreatic cancer cell lines PANC-1, PK-1, and PK45H were cultured with MK615 at concentrations of 600, 300, 150, and 0 μg/mL. Growth inhibition was evaluated by cell proliferation assay, and killing activity was determined by lactate dehydrogenase (LDH) assay. Expression of Aurora A and B kinases was detected by real-time polymerase chain reaction (PCR) and Western blotting. Cell cycle stages were evaluated by flow cytometry. RESULTS: The growth inhibitory rates of MK615 at 150, 300, and 600 μg/mL were 2.3% ± 0.9%, 8.9% ± 3.2% and 67.1% ± 8.1% on PANC1 cells, 1.3% ± 0.3%, 8.7% ± 4.1% and 45.7 ± 7.6% on PK1 cells, and 1.2 ± 0.8%, 9.1% ± 2.1% and 52.1% ± 5.5% on PK45H cells, respectively (P <0.05). The percentage cytotoxicities of MK615 at 0, 150, 300, and 600 μg/mL were 19.6% ± 1.3%, 26.7% ± 1.8%, 25.5% ± 0.9% and 26.4% ± 0.9% in PANC1 cells, 19.7% ± 1.3%, 24.7% ± 0.8%, 25.9% ± 0.9% and 29.9% ± 1.1% in PK1 cells, and 28.0% ± 0.9%, 31.2% ± 0.9%, 30.4% ± 1.1% and 35.3 ± 1.0% in PK45H cells, respectively (P < 0.05). Real-time PCR and Western blotting showed that MK615 dually inhibited the expression of Aurora A and B kinases. Cell cycle analysis revealed that MK615 increased the population of cells in G2/M phase. CONCLUSION: MK615 exerts an anti-neoplastic effect on human pancreatic cancer cells in vitro by dual inhibition of Aurora A and B kinases.

关键词： Japanese apricot Aurora kinases Pancreatic cancer Cell cycle

New anti-proliferative agent,MK615,from Japanese apricot “Prunus mume” induces striking autophagy in colon cancer cells in vitro

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World Journal of Gastroenterology 2007年第48期13卷 6512-6517页

作者： Shozo Mori Tokihiko Sawada Toshie Okada Tatsushi Ohsawa Masakazu Adachi kubota keiichi Second Department of Surgery Dokkyo University School of Medicine Japan Apricot Co. Ltd

AIM: To investigate the anti-neoplastic effects of MK615, an extract from the Japanese apricot (Prunus mume), against colon cancer cells. METHODS: Three colon cancer cell lines, SW480, COLO, and WiDr, were cultured with MK615. Growth inhibition was evaluated by cell proliferation assay and killing activity was determined by lactate dehydrogenase assay. Induction of apoptosis was evaluated by annexin Ⅴ flow cytometry. Morphological changes were studied by light and electron microscopy, and immunofluorescence staining with Atg8. RESULTS: MK615 inhibited growth and lysed SW480, COLO and WiDr cells in a dose-dependent manner. Annexin Ⅴ flow cytometry showed that MK615 induced apoptosis after 6 h incubation, at which point the occurrence of apoptotic cells was 68.0%, 65.7% and 64.7% for SW480, COLO, and WiDr cells, respectively. Light and electron microscopy, and immunofluorescence staining with Atg8 revealed that MK615 induced massive cytoplasmic vacuoles (autophagosomes) in all three cell lines. CONCLUSION: MK615 has an anti-neoplastic effect against colon cancer cells. The effect may be exerted by induction of apoptosis and autophagy.

关键词： Colon cancer Japanese apricot Prunus mume Autophagy Apoptosis MK615

Rectal angiolipoma diagnosed after surgical resection: A case report

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World Journal of Gastroenterology 2007年第3期13卷 467-469页

作者： Mitsuru Ishizuka Hitoshi Nagata Kazutoshi Takagi Toru Horie Akihito Abe keiichi kubota Department of Gastroenterological Surgery Dokkyo University School of Medicine Tochigi Japan

Angiolipoma is a common benign tumor with a char- acteristic vascular component that usually occurs in subcutaneous tissue. Although lipomas are frequently encountered at colonoscopy as submucosal tumors, an- giolipomas are rarely found in the gastrointestinal tract including the large intestine. Here we report a 77-year old Japanese man who underwent transanal resection of a tumor that was diagnosed tentatively as a leiomyoma. Histologically, the tumor consisted of mature fat cells and blood vessels. Immunohistochemically, the tumor cells were negative for c-kit and HMB-45, which are consis- tently expressed in tumors such as gastrointestinal stro- mal tumor and angiomyolipoma. The tumor was there- fore diagnosed as an angiolipoma. This is thought to be the first case report of a surgically resected angiolipoma of the rectum.

关键词： Angiolipoma Angiomyolipoma Gastrointestinal stromal tumor

Intraoperative frozen section diagnosis of bile duct margin for extrahepatic cholangiocarcinoma

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World Journal of Gastroenterology 2018年第12期24卷 1332-1342页

作者： Takayuki Shiraki Hajime Kuroda Atsuko Takada Yoshimasa Nakazato keiichi kubota Yasuo Imai Department of Gastroen- terological Surgery Dokkyo Medical University Department of Diagnostic Pathology Dokkyo Medical University

AIM To evaluate the usefulness of frozen section diagnosis(FSD) of bile duct margins during surgery for extrahepatic cholangiocarcinoma(CCA). METHODS We retrospectively analyzed 74 consecutive patients who underwent surgery for extrahepatic CCA from 2012 to 2017, during which FSD of bile duct margins was performed. They consisted of 40 distant and 34 perihilar CCAs(45 and 55 bile duct margins, respectively). The diagnosis was classified into three categories: negative, borderline(biliary intraepithelial neoplasia-1 and 2, and indefinite for neoplasia), or positive. FSD in the epithelial layer, subepithelial layer, and total layer was compared with corresponding permanent section diagnosis(PSD) ***, association between FSD and local recurrence was analyzed with special reference to *** Analysis of 100 duct margins revealed that concordance rate between FSD and PSD was 68.0% in the total layer, 69.0% in the epithelial layer, and 98.0% in the subepithelial layer. The extent of remaining biliary epithelium was comparable between FSD and PSD, and more than half of the margins lost > 50% of the entire epithelium, suggesting low quality of the samples. In FSD, the rate of negative margins decreased and that of borderline and positive margins increased according to the extent of the remaining epithelium. Diagnostic discordance between FSD and PSD was observed in 31 epithelial layers and two subepithelial layers. Alteration from borderline to negative was the most frequent(20 of the 31 epithelial layers). Patients with positive margin in the total and epithelial layers by FSD demonstrated a significantly worse local recurrence-free survival(RFS) compared with patients with borderline and negative margins, which revealed comparable local RFS. Patients with borderline and negative margins in the epithelial layer by PSD also revealed comparable local RFS. These results suggested that epithelial borderline might be regarded substantially as negati

关键词： Cholangiocarcinoma Bile duct cancer Frozen section diagnosis Permanent section diagnosis Bile duct margin Biliary intraepithelial neoplasia Dysplasia Indefinite for neoplasia Borderline lesion Local recurrence

Intraductal papillary neoplasm of the bile duct developing in a patient with primary sclerosing cholangitis: A case report

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World Journal of Gastroenterology 2014年第42期20卷 15925-15930页

作者： Hiroyuki Hachiya Junji Kita Takayuki Shiraki Yukihiro Iso Mitsugi Shimoda keiichi kubota Second Department of Surgery Dokkyo Medical University

We report a case of intraductal papillary neoplasm of the bile duct(IPNB) that developed in a patient with primary sclerosing cholangitis. A 46-year-old woman was admitted to our hospital with obstructive jaundice. The liver function tests demonstrated increased serum liver enzyme levels. Computed tomography showed dilatation of the intrahepatic bile ducts. Abdominal ultrasonography revealed a highly echoic protruding lesion in the posterior bile duct near the right lobe of the liver. The lesion was suspected to be IPNB, but we were unable to confirm whether it was a carcinoma. A right hepatectomy was performed, and this showed that the dilated bile duct was filled with mucin and contained several yellowish papillary tumors. Histologically, the neoplastic biliary epithelium showed papillary growth in the dilated lumen. The tumor was diagnosed as IPNB, high-grade intraepithelial neoplasia secreting abundant mucin. No recurrence has been detected 3 years after surgery.

关键词： Intraductal papillary neoplasm of the bile duct Primary sclerosing cholangitis Cholangiocarcinoma High-grade intraepithelial neoplasia Premalignant lesion