RESUMEN
To enrich the resource pool of endophytic fungi from plants which produce taxol, a taxol-producing endophytic fungus TMS-26 was isolated from the stem of Taxus Media. The result of high performance liquid chromatography (HPLC) showed that TMS-26 extract exhibited similar chromatographic peaks and retention time (4.545 min) with authentic taxol. Then mass spectrometry (MS) analysis further confirmed that TMS-26 extracts contained the same mass peaks with authentic taxol ((M+Na)+=876). These indicated that the isolated endophytic fungus TMS-26 can produce taxol. According to the morphological characteristics, the molecular analysis of 18S rDNA and internal transcribed spacer nuclear rDNA gene sequence, the fungus was identified as Aspergillus fumigatus TMS-26.
RESUMEN
<p><b>OBJECTIVE</b>To observe human tumor necrosis factor-alpha (hTNF-alpha) expression and secreting level of human embryo myoblasts transfected by hTNF-alpha gene.</p><p><b>METHODS</b>Human embryo myoblasts were transfected with shuttle plasmid pSV23SHTNF containing hTNF-alpha gene by cationic liposomes DOSPER. The control group was only given equivalent liposomes except plasmid. After culturing for 24, 48, 72 and 96 hours, hTNF-alpha expression level of human embryo myoblasts was observed with immunocytochemistry staining, and hTNF-alpha secreting of human embryo myoblasts was analyzed by ELISA.</p><p><b>RESULTS</b>After transfected by hTNF-alpha gene for 24, 48, 72 and 96 hours, the human embryo myoblasts displayed significant secretion of hTNF-alpha in the cultural supernatant (P < 0.05), and overexpression in cytoplasma and cell membrane.</p><p><b>CONCLUSION</b>Transfection of hTNF-alpha gene to human myoblasts made myoblasts secrete high concentration of hTNF-alpha, implying it is feasible that transfecting muscle cells surrounding tongue carcinoma lesion with hTNF-alpha gene can prevent tongue carcinoma from intruding into deeper muscle tissue.</p>