Separation and identification of Taxol in the crude extracts of Taxus cuspidata and its callus culture with HPLC-ESI-MS/MS / 药学学报

<p><b>AIM</b>To study the MS/MS fragmentation mechanism of Taxol, and based on it to establish HPLC-ESI-MS/MS technique to separate and identify Taxol in the crude extracts of Taxus cuspidata and its callus culture, consequently to provide a fast and credible method for the analysis of Taxol in natural products.</p><p><b>METHODS</b>Optimized the HPLC-ESI-MS/MS parameters for the sample analysis, and then discussed the ionization and cleavage mechanism of Taxol in ESI-MS and ESI-MS/MS, finally identified the Taxol in the samples with retention time, molecular weight and MS/MS spectra.</p><p><b>RESULTS</b>Elucidated the MS/MS fragmentation mechanism of Taxol, and developed HPLC-ESI-MS/MS method to analyze Taxol in the two samples.</p><p><b>CONCLUSION</b>The HPLC-ESI-MS/MS method is rapid, highly sensitive and specific, so it is suitable for the separation and identification of Taxol in natural products.</p>

Effect of replicating adenovirus harboring p53 gene on chemosensitivity of hepatocarcinoma cell line / 第二军医大学学报

Objective: To construct a replicating adenovirus vector CNHK600-p53 carrying p53 gene and investigate its effect on the chemosensitivity of hepatocarcinoma cell line. Methods: Chemotherapy of hepatocarcinoma cel ls BEL-7404 were carried out using Fluorouracil, Mitomycin, Epirubicin, CNHK600-p53, CNHK600-p53 + Fluorouracil, CNHK600-p53 + Mitomycin, and CNHK600-p53 + Epirubicin, separately. MTT assay was used to evaluate the killing effects after therapy. Results: The inhibition rate on BEL-7404 cells was(47±3)% whe n using 100 μg/ml 5-Fu, was (20±4)% when using 1 μg/ml MMC, and was (73±2)% when using 2.5 μg/ml EPI. The inhibition rates of BEL-740 cells in CNHK600-p53 (MOI=10) + Fluorouracil (100 μg/ml), CNHK600-p53 (MOI=10) + Mitomycin (1 μg/ml), and CNHK600-p53 (MOI=10) + Epirubicin (2.5 μg/ml) groups were (59±4)%, (44±4)% and (86±2)%, respectively (P<0.01). Conclusion: Adenovirus CNHK600-p53 carrying p53 gene can enhance chemosensitivity of BEL7404 hepatocarcinoma cell line. Gene therapy using adenovirus CNHK600-p53 in combination with chemotherapy may be a new strategy for hepatocarcinoma treatment.