| Project/Area Number |
60570297
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
内科学一般
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| Research Institution | Kagoshima University |
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Principal Investigator |
HIRONO Shuichi (1986) Department of Biochemistry, Kagoshima University Dental School., 歯学部, 助手 (00181222)
坪内 博仁 鹿児島大学, 医学部, 助手
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| Co-Investigator(Kenkyū-buntansha) |
NAKAYAMA Hiroyuki Department of Biochemistry, Kagoshima University Dental School., 歯学部口腔生化学, 助手 (00172479)
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| Project Period (FY) |
1985 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1986: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1985: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Hepatotrophic Factor / Fulminant Hepatic Failure / Primary Culture of Adult Rat Hepatocytes / DNA Synthesis / Human Hepatocyte Growth Factor / hHGF / 精製 |
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| Research Abstract |
According to the initial plan, we performed the study on the human hepatocyte growth factor(hHGF) and obtained the following results.
(1)The effect of sera from patients with various liver diseases on liver cell growth was investigated by use of adult rat liver cells in primary culture. Sera from patients with chronic hepatitis or liver cirrhosis had no or slight growth- promoting activity, respectively, compared with that of normal human sera. Sixteen of seventeen sera from patients with fulminant hepatic failure showed marked stimulatory activity on DNA synthesis of about 16 times an average value of normal human sera. The growth- promoting activity was confirmed by showing that the patients' serum increased the labeling index with [ <^3H> ]thymidine and total number of nuclei in hepatocyte cultures.
(2)Appearance of hHGF in sera of patents with fulminant hepatic failure seems to be related to their coma grade. Its level increased when the patient fell into a state of more advanced coma and decreased immediately in the recovery stage.
(3)Plasma from patients with fulminant hepatic failure obtained during plasma exchange therapy, like their serum,demonstrated marked stimulatory activity on DNA, synthesis in cultured rat hepatocytes. The activity of pooled active fractions, obtained by gel filtration of the heated plasma, was lost completely on heat treatment at 80 ゜C for 10 min. or on treatment with trypsin or chymotrypsin, which suggests that it was due to a protein.
HHGF was purified about 10,000-fold from heated plasma of a patient by ammonium sulfate precipitation and chromatographies on Affi-Gel Blue and heparin-Sepharose. The molecular weight of hHGF was about 85,000 as determined by SDS-PAGE.
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