原発性胆汁性肝硬変の動物モデルの作製: Methylene dianilineのマウスへの長期投与の検討

金沢大学医薬保健研究域医学系Methylene dianiline(MDA)を用いた原発性胆汁性肝硬変の動物モデルの作製I.予備実験1.投与量の決定肝内外の胆道系上皮に障害を惹起するMDAの最小有効量を決定するために,種々の濃度のMDAをマウス(BALB/c,雌,4週齢)に腹腔内投与し,検討した結果,至適投与量は20.0mg/kg(約0.4mg/mouse)であることが判明した.2.ヒトPDC-E2の合成及び精製米国カリフォルニア大学デービス校Gershwin教授より分与されたヒトPDC-E2のDNAを組み込んだ大腸菌を大量培養して,ヒトPDC-E2を発現させ,種々の操作により精製し,回収した.II.本実験予備実験で得られたMDA投与量をBAL/cマウスの(雌,4週齢)を用いて,一年間に渡る実験を遂行した.MDAは2カ月毎に腹腔内に投与し,また,ヒトPDC-E2の免疫は1カ月毎に施行した.コントロール群として,MDA不投与群,ヒトPDC-E2の代替としてウシ・アルブミンで免疫した群を設け,具体的には以下の4群に分けて実験を実施した.なお,各グループのマウスは2カ月毎に数匹ずつ犠死させ,臓器と血清を採取した.グループ1 MDA投与+ヒトPDC-E2免疫 グループ3 MDA不投与+ヒトPDC-E2免疫グループ2 MDA投与+ウシ・アルブミン免疫 グループ4 MDA不投与+ウシ・アルブミン免疫本実験を一年間に渡って遂行したが,ヒトPDC-E2を免疫したグループ群では血清学的にヒトPDC-E2に対する抗体が100%に頻度で産生されていることが示された.さらにマウスのPDC-E2に対する自己抗体も半数以上のマウスで出現することが判明した.ただし,肝臓の形態学的観察ではMDAの投与により一過性に胆管が障害されるものの,慢性胆管炎が遷延性に持続することはなかった.研究課題/領域番号:11770089, 研究期間(年度):1999 – 2000出典：「原発性胆汁性肝硬変の動物モデルの作製: Methylene dianilineのマウスへの長期投与の検討」研究成果報告書　課題番号11770089（KAKEN：科学研究費助成事業データベース（国立情報学研究所）） （ https://kaken.nii.ac.jp/ja/grant/KAKENHI-PROJECT-11770089/ ）を加工して作

マウス胆道系上皮細胞の区分別培養法の確立

取得学位 : 博士（医学）, 学位授与番号 : 医博甲第1200号, 学位授与年月日：平成8年3月25日,学位授与年：199

A Case of Hypereosinophilia-Associated Multiple Mass Lesions of Liver Showing Non-Granulomatous Eosinophilic Hepatic Necrosis

Hypereosinophilic syndrome (HES) is defi ned by elevation more than 1.5×109/L of presence of a peripheral blood count, evidence of organ involvement, and exclusion of secondary eosinophilia such as allergic, vasculitis, drugs, or parasite infection and also clonal eosinophilia. We present the HES case with hepatic involvement. The patient is 70-year-old male. He complained fever and back pain. Blood examination showed marked peripheral eosinophilia, elevation of transaminase and biliary enzymes. Multiple irregular mass lesions of the liver were pointed out by CT and MRI. The liver biopsy was done for differentiation from malignancy. In parenchyma, hepatic necrotic lesion was observed accompanying severe eosinophilic infi ltration with Charcot-Leyden’s crystals. There was granulomatous reaction. He was diagnosed as HES and got recovery due to steroid therapy. From the review of HES article, the hepatic histology is categorized into four types as below: 1) cholangitis type; 2) chronic active hepatitis type; 3) vasculopathic type, 4) hepatic necrosis type. Our case is classifi ed in hepatic necrosis type. This type seems to be important to distinguish malignant tumor and also visceral larva migrans by liver biopsy

A fatal case of progressive steatohepatitis, possibly chemotherapy- associated steatohepatitis related to gemcitabine

金沢大学医薬保健研究域医学系We report the case of an 80-year-old female suffering from pancreatic cancer who developed severe non-alcoholic steatohepatitis (NASH) resulting in fatal hepatic failure after anti-cancer chemotherapy with gemcitabine. Hepatic encephalopathy appeared 1 year after the chemotherapy, and the patient developed progressive liver failure and eventually died. Radiological examination showed severe fatty liver. Histopathological examination of a liver needle necropsy showed almost panlobular macrovesicular fatty change. Ballooning degeneration and necrosis of hepatocytes accompanying neutrophil infiltration, Mallory bodies, and a few bile plugs were found in zone 3. Marked perivenular and pericellular/perisinusoidal fibrosis and extensive bridging fibrosis were also found. Together, these findings indicated steatohepatitis at a precirrhotic stage. Because the patient had no history of drinking in excess, we made a diagnosis of NASH, in particular, chemotherapy-associated steatohepatitis (CASH). Gemcitabine is a pyrimidine nucleoside antimetabolite with anti-cancer activity. A few reports have mentioned fatal hepatotoxicity caused by gemcitabine, but, to our knowledge, this is the first report of steatohepatitis, possibly associated with gemcitabine. Physicians treating patients with this drug should be aware of the possibility of steatohepatitis. © Springer 2010

The case of double primary lung adenocarcinomas with an EGFR mutation and ALK translocation successfully treated with alectinib at the post-surgicalrecurrence

A 36-year-old male was found two nodules in the right lower lobe of the lung. After the surgical resection, both lesions were diagnosed as invasive adenocarcinomas. One lesion was primarily lepidic growth component with EGFR-L858R mutation, and the other was micropapillary component with ALK translocation accompanying mediastinal lymphnode metastases. While he experienced disease recurrence, the disease was controlled by an ALK inhibitor, given based on the findings of surgical specimens. This is the first case who had two simultaneous lung cancers with EGFR mutation and ALK translocation in each respective lesion, and was successfully treated with ALK inhibitor at the post-surgical recurrence

Clinicopathological significance of antinuclear antibodies in non-alcoholic steatohepatitis

金沢大学大学院医学系研究科がん細胞学Aim: Serum antinuclear antibodies (ANA) are occasionally noted in patients with non-alcoholic steatohepatitis (NASH). We examined the significance of ANA in NASH. Methods: We compared clinicopathological features in patients with ANA-positive NASH (n = 35) and ANA-negative NASH (n = 36). Inflammatory cell profiles and the distribution of oxidative stress markers were also examined immunohistochemically. Results: ANA-positive NASH was significantly associated with female gender (P = 0.005), high degree of portal inflammation (P = 0.039), interface activity (P = 0.036) and hepatocellular ballooning (P = 0.0008). In addition, ANA of high titer (320-fold or more) was significantly associated with the histological grade and stage of NASH (P = 0.02). The degree of steatosis wais rather mild in the high-titer ANA group(P = 0.01). The analysis of inflammatory cell profiles revealed that CD3-positive T cells were predominant and plasma cells were rather few in the portal area and hepatic lobules in both ANA-positive and ANA-negative groups. There was no difference in the distribution of oxidative stress markers between ANA-positive and ANA-negative groups. Conclusion: These findings suggest that the presence of ANA may be related to the progression of NASH and that a different type of autoimmune mechanism may be involved in the pathogenesis of NASH with ANA, compared to the pathogenesis of autoimmune hepatitis. © 2007 The Japan Society of Hepatology