Objective To establish an HPLC method for the simultaneous determination of the 6 aconitum alkaloids in compoundTengwu Ointment.Methods Chromatographic column was Aglient XDB-C18 column (4.6 mm × 250 mm, 5μm), with the mobile phase A of acetonitrile- tetrahydrofuran (25∶8), phase B was 0.1 mol/L ammonium acetate solution (0.5 mL acetic acid added to the 1000 mL) with gradient elution;the flow rate was 1 mL/min;the detection wavelength was set at 235 nm;the column temperature was 25℃.Results Aconitine was in the good linear range of 0.072-0.648μg (r=0.999 5), with the average recovery of 99.29%, RSD=1.25%. Mesaconitine was in the good linear range of 0.062-0.648μg (r=0.999 5), with the average recovery of 99.12%, RSD=0.85%. Hypaconitine was in the good linear range of 0.064-0.384μg (r=0.999 8), with the average recovery of 99.57%, RSD=1.07%. Benzoylaconine was in the good linear range of 0.056-0.672μg (r=0.999 2), with the average recovery of 98.11%, RSD=0.61%. Benzoylmesaconine was in the good linear range of 0.055-0.993μg (r=0.999 9), with the average recovery of 99.27%, RSD=1.10%. Benzoylhypaconine was in the good linear range of 0.078-0.702μg (r=0.999 8), with the average recovery of 99.08%, RSD=1.38%.Conclusion This method is simple, sensitive, repeatable and accurate, which can be used for determination of aconitum alkaloids in compoundTengwu Ointment.

Objective To investigate the expression of heparanase in non-melanoma skin cancer including cutaneous squamous cell carcinoma (CSCC) and basal cell carcinoma (BCC) and the relationship among heparanase expression and pathologic grading and lymph nodes metastasis of CSCC. Methods The expressions of heparanase were detected by RT-PCR assay and immunohistochemical staining in 38 cases of CSCC, 33 BCC and 19 normal human skin tissues. Then the relationship between heparanase expression and CSCC clinical pathological features was explored. Results The positive rate ( 68.4%, 60. 5% ) of heparanase mRNA and heparanase in CSCC (24. 2%, 18. 2% ) was significantly higher than those of BCC and normal skin tissue ( 10. 5% ,5.3% ) ( P <0. 01 ) . Heparanase expression rate in CSCC with lymph nodes metastasis (92. 3% ,84.6% ) and Ⅲ～Ⅳ(Broders) (92. 3% ,84. 6% ) was higher than those in tumor without lymph nodes metastasis (56. 0% ,48.0% ) and Ⅰ～Ⅱ (Broders) (56. 0% ,48. 0% ) ( P <0. 05). Conclusions Heparanase was over-expressed in CSCC compared with BCC and normal skin tissues. Expression of heparanase was correlated to pathologic grade and lymph nodes metastasis of CSCC.