BACKGROUND: Talomerase activity inhibitor inhibits or kills renal carcinoma cells, and also affects stem cells that play importan roles in occurrence and development of renal carcinoma.OBJECTIVE: To observe renal carcinoma stem cell surface marker CD133 and telomerase activity expression in serum-free suspension culture, and to compare with renal carcinoma cells in serum suspension culture.DESIGN, TIME AND SETTING: The in vitro cytological study was performed at the Jiangsu University from June 2008 to Februar 2009.WIATERIALS: Fresh normal renal tissue surrounding renal carcinoma was obtained from Affiliated Hospital, Jiangsu University.Renal carcinoma stem cell line OS-RC-2 was supplied by Cell Bank, Chinese Academy of Sciences Shanghai Branch.METHODS: OS-RC-2 in logarithmic phase, digested by trypsin, and centrifuged. Supematant was removed. OS-RC-2 cell line in serum-free DMEM/F12 supplemented with epidermal growth factor and basic fibroblast growth factor was incubated at 2×105/L in 5% CO2 incubator at 37℃. Renal carcinoma cultured in serum and normal renal tissue served as controls.MAIN OUTCOME MEASURES: Cell growth was observed under an inverted microscope. Expression of CD133 and CD34 was detected using flow cytometry. Reel-time quantitative TRAP assay was applied to evaluate telomerase activity in renal carcinoma stem cells.RESULTS: After incubated in serum-free medium, renal carcinoma stem cells were round and suspended. Two days later, cell mass generated. Each cell mass contained 3-8 cells, with strong refraction. Seven days later, cell mass became more, presented big body that was regular, round or elliptical. CD133+CD34- rate in renal carcinoma stern cell mass was significantly greater in serum-free suspension culture compared with in serum suspension culture. CD133 and CD34 expression was not determined in normal renal tissue. There were significant differences among groups (F=328.25, P < 0.05). Telomerase activity was greater in renal carcinoma stem cells and renal carcinoma cells compared with normal renal ceils (F=-278.74, P < 0.05). No significant difference was detected between renal carcinoma stem cells and renal carcinoma cells.CONCLUSION: Compared with serum cultured renal carcinoma cells, serum-free cultured renal carcinoma cell surface marker CD133 presents high expression. Moreover, talomerase activity is high in renal carcinoma stem cells and renal carcinoma cells compared with normal renal tissue.

OBJECTIVETo develop an HPLC method for content determination of protocatechuic acid, chlorogenic acid and caffeic acid in Zhuang medicine Blumea megacephala (Randeria), and explore the content variation of the 3 components of the herbs harvesting in different months, and provide the scientific basis of reasonable application.

METHODThe determination was carried out on a Shimadzu VP-ODS column (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile and water containing 0.1% phosphoric acid in a linear gradient elution. The flow rate was 1 mL min(-1), and the detected wavelength was set at 258, 327nm.

RESULTThe peak areas and the concentrations of the three components had good linear relationship in the range of 1.7-17 mg x L(-1) for protocatechuic acid, 15.6-156 mg x L(-1) for chlorogenic acid, 3.96-39.6 mg x L(-1) for caffeic acid. The average recoveries were 103.4%, 102.2%, 98.5%, respectively.

CONCLUSIONThe method was proved to be simple, accurate and used for the quality evaluation of Blumea megacephala (Randeria).

Asteraceae ; chemistry ; Caffeic Acids ; chemistry ; Chlorogenic Acid ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Hydroxybenzoates ; chemistry ; Plants, Medicinal ; chemistry