The AcOEt eluate (45.8 g) was repeatedly chromatographed over silica gel with CHC13-MeOH and benzene-AcOEt to give betulonic acid as colorless powder (424 mg). Determination of antihepatotoxicity Assays using CCI4-and GaIN-induced cytotoxicity in primary cultured rat hepatocytes in vitro were carried out as described previously (11, 12).
New 5-nucleotidase inhibitors, NPF-861A, NPF-861B, NPF-8611A, and NPF-8611B were isolated from a plant origin. According to their physico-chemical properties, these compounds are polyphenolic substances. These inhibitors displayed significant therapeutic activity by intraperitoneally administration against intraperitoneally inplanted Ehrlich ascites carcinoma. In vitro, these inhibitors exhibited a moderate cytotoxicity to Ehrlich ascites carcinoma strain E, HeLa, and HL-60 cells. However, these 5'-nucleotidase inhibitors did not show an anti-tumor effects on L1210 cells in vivo and in Vitro.
We have constructed a new cloning vector, pYUM1118, for generating deletions. Recently, the kilo-base sequencing method, which uses exonuclease III (ExoIII) and ss DNA specific nuclease, is widely used as a rapid sequencing method of long DNA fragment (>1000 bp) (1). To employ this method, it is necessary to clone the target DNA into the polylinker site of the vectors such as pUC plasmids or M13mp RFs. However,
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