OBJECTIVE:To establish a determination method for trace elements of Plumbago zeylanica. METHODS:Induct- ively coupled plasma emission spectrum (ICP-AES) was used to determine seven trace elements of P. zeylanica, i.e. Cu, Zn,Fe,Ca,Mg,Mn and Sr simultaneously. Se was determined by atomic fluorescence (AFS) method. RESULTS: The RSD ranged from 0.78% to 3.01%(n=5),while the recovery was within 91.9%～105.8%. The result obtained was satisfactory. Se took a large proportion in P. zeylanica, followed by Cu,Zn,Fe,Ca,Mg,Mn,Sr.CONCLUSION: The method is brief, rapid and accurate for the determination of P. zeylanica. The study provide reference for future pharmacological study of P. zeylanica.

Objective To study the effects of the gingerol on the melanogenesis in melanoma B16 cells. Methods Melanoma cells were cultured with gingerol at 12. 5, 25. 0, 50. 0, 100. 0, 200. 0 μmol · L-1 and positive control drug hydroquinone,respectively,using Dulbecco's modified eagle's medium(DMEM) as the blank control group. The cell proliferation was measured by methyl thiazolyltet tetrazolium ( MTT) colorimetric assay. The tyrosinase activity and melanin content were measured by colorimetry assay. Results Gingerol at different concentrations had inhibitory effect on B16 cell proliferation compared with the blank control group ( P < 0. 05 or P < 0. 01), the inhibition rate being more than 48% at the dosage of 200. 0 μmol·L-1 . Tyrosinase activity was inhibited significantly compared with blank control group(P<0. 05 or P<0. 01),the inhibition rate being up to 50% at the dosage of 200 μmol·L-1 . Melanin content was also decreased at all levels of gingerol compared with blank control group(P<0. 05 or P<0. 01),but not in a dose-dependent manner. The inhibition rate of melanin content reached the plateau at gingerol levels greater than 25. 0 μmol · L-1 . Conclusion Gingerol can inhibit the cell proliferation,tyrosinase activity and decrease melanin synthesis in certain range of concentrations.

Objective To sieve the bioactive constituents of Plumbago zeylanica, serum pharmacochemistry research was performed. Method Based on the establishment of HPLC fingerprints of Plumbago zeylanica, the constituents absorbed into blood were determined by comparing the HPLC fingerprints of the methanol extracts, medicated serum samples and blank serum sample. Results One compound absorbed into blood was detected, the other might be metabolites of the original constituent. Conclusion The one onstituent absorbed into blood was possible bioactive component of Plumbago zeylanica.

ObjectiveTo investigate the effects of extracts ofLiaoxuan Kaxifu Powders (LE) on proliferation and transcription of IL-8 and ICAM-1 in HaCaT induced by TNF-α; To discuss its mechanism of action.Methods Cultured HaCaT was assigned into normal group, TNF-α group and low-, medium- and high-dose of LE group. Every group was induced by 40 ng/mL TNF-α except for normal group, and then LE groups were treated by different concentrations (8, 40, 200μg/mL) of LE for 48 h. The proliferation of HaCaT was evaluated by MTT and the apoptosis was detected by inverted fluorescence microscope. Levels of IL-8 and ICAM-1 in HaCaT were assessed by ELISA, and their mRNA expressions was detected by semi-quantitative RT-PCR.Results Compared with normal group, the absorbency of HaCaT and the contents and mRNA expressions of IL-8 and ICAM-1 increased in TNF-α group (P<0.05,P<0.01); compared with TNF-α group, LE of all dose groups could significantly inhibit the absorbency, decrease the contents and mRNA expressions of IL-8 and ICAM-1 (P<0.05,P<0.01).Conclusion LE is able to inhibit the proliferation of HaCaT induced by TNF-α, and the mechanism is probably related to promoting apoptosis and down-regulating the gene expressions of IL-8 and ICAM-1, and then maintaining the normal level of HaCaT.

Objective:Vernonia anthelmintica,with complicated components,is a common medicine for the treatment of vitiligo.The purpose of this study was to establish and optimize the separation conditions of high performance liquid chromatography fingerprints(HPLC-FPs) for Vernonia anthelmintica(L.) Willd.Methods: We investigated the HPLC-FPs of different extraction samples by the retention times and relative areas of common peaks in the fingerprints.Results: We established the optimum separation conditions of HPLC-FPs for Vernonia anthelmintica(L.) Willd,and obtained 19 common peaks in the fingerprints.The HPLC-FPs of the 85% ethanol extract were quite similar to those of the semi-bionic extract(SBE) but very different from those of the water extract of Vernonia anthelmintica(L.) Willd.The No.1,4,6,11,12,14 and 16-19 peaks were the endemic ones of the HPLC-FPs of the 85% ethanol extract and SBE of Vernonia anthelmintica(L.) Willd,while No.20-30 were the endemic peaks of the HPLC-FPs of the water extract.Conclusion: The results of our study has provided experimental evidence for further investigation into the constituents absorbed into blood,pharmaceutical technology and pharmacodynamics of Vernonia anthelmintica(L.) Willd.

OBJECTIVE:To study the extraction rate and speciation of antivitiligo-related elements in Opercalina turperthum by water extraction and semi-bionic extraction. METHODS:Water extraction and semi-bionic extraction were respectively used. Wa-ter-soluble state and suspension state in extract of O. turperthum were separated by microporous filtering film;organic and inorgan-ic trace elements in water-soluble state were separated by macroporous resin. The contents of Cu,Zn,Fe,Ca,Mg,Mn and Sr were detected by flame atomic adsorption spectrophotometry,and speciation analysis was conducted. RESULTS:After water extrac-tion,extraction rates of 7 trace elements were 40.47%-72.49%;ratio of suspended particles was 3.69%-8.78%;ratio of organic state/inorganic state was 104.36% in water-soluble state of Sr and 3.94%-48.39% in water-soluble state of Cu,Zn,Fe,Ca,Mg, Mn. After semi-bionic extraction,except for Mn,extraction rates of trace elements were higher than water extraction,extraction rates of Cu,Zn,Fe,Ca,Mg,Sr were 77.69%-90.19%;ratio of suspended particles was 0.39%-8.57%;the ratio of organic state/inorganic state was 72.74%-180.79% in water-soluble state of elements. CONCLUSIONS:Cu,Zn,Fe,Ca,Mg,Mn and Sr in O. turperthum are dissolved easily,mainly existing in the form of inorganic state. After semi-bionic extraction,except for Mn,the dis-solution rate of other elements and the proportion of organic trace elements after dissolution increase significantly. Both dissolution and effective utilization rate of trace elements by semi-bionic extraction are higher than water extraction.

Objective To optimize the best condition of semi-bionic extraction for Vernonia anthelmintica.Methods The best condition of the semi-bionic extraction for V.anthelmintica by uniform design was optimized with total HPLC integrate area of common peaks in fingerprints,total flavonoids and dried extract weight as the indexes.Results According to industry production condition,the best technical condition:pH values of 80% ethanol for the 1st,2nd,and 3rd decoctions were in order of 6.0,6.5,and 8.0,and the extraction time were 1,0.5,and 0.5 h,respectively.Conclusion It is better for V.anthelmintica extracted by semi-bionic extraction,and the parametrs are optimized by uniform design.

OBJECTIVE:To investigate the in vitro transdermal absorption properties of galangin and effects of different pene-tration enhancers on its transdermal behaviors,and provide reference for developing skin preparations using galangin as APIs in the treatment of vitiligo. METHODS:HPLC was used to determine the galangin content. Using cumulative permeation rate (Q) and the transdermal rate(J)of galangin as indexes,the effect of absorption of receiving solution [20%,40% polyethylene glycol 400 (PEG400)solution and 30% ethanol solution] and rotating rate(200,300,400 r/min)on galangin in complete skin of mice were investigated,as well as the azone(1%,3%,5%)and propylene glycol(10%,20%,40%)alone or combination on its penetra-tion promotion. And the transdermal properties of galangin in complete skin,exfoliating skin,dermis skin of rats and mice were de-tected. RESULTS:The best permeability of complete skin of mice showed in 40% PEG400 solution at rotating speed of 300 r/min with 5% azone alone,J was 3.2570 μg/(cm2·h). Js of complete skin,exfoliating skin,dermis skin of mice were 2.7199,34.016, 33.874 μg/(cm2·h),respectively;and those of rats were 0.4996,9.5124,17.406 μg/(cm2·h). CONCLUSIONS:Galangin can penetrate the complete skin of mice and rats,however,the penetration quantity is far lower than exfoliating skin and dermis skin.

Objective To investigate the effect of acteoside on injury PC12 cells induced by glutamate. Methods PC12 cells were assigned into normal control group, model control group, positive drug group and acteoside(AS) treated group. Every group was treated by 1. 5 mmol·L-1 glutamate for 24 hours except for the control group, and the injury was antagonized by 10 μmol·L-1 Vit E and acteoside at different concentration(15. 625, 31. 25, 62. 5, 125 and 250 μmol·L-1 ). Cell morphology was observed by inverted microscope, cell survival was determined with MTT, LDH activity was measured by enzyme label kit, the MDA content and SOD activity were measured by TBA kit and WST kit, and the ROS was measured by Elisa kit. Results Compared with the model control group, all doses of acteoside could significantly improve the PC12 cell morphology and survival (P<0. 05), inhibit LDH activity and production of MDA and ROS (P<0. 05), increase the activity of SOD (P<0. 05), except for the lowest dose group. Conclusion Acteoside has protective effects on PC12 cells injured by glutamate.

Objective To investigate the effect of Galangin of different purity on melanocyte proliferation and melanin synthesis of A375-HaCaT co-culture system. Methods The A375-HaCaT co-culture system was established with Transwell technology, and 0.1, 0.5, 1.0, 5.0, 10 μg/mL Galangin of the purity of 70%and 90%was used to act on the co-culture system. Cell proliferation, melanin content and tyrosinase of A375 were measured by MTT assay, NaOH lysis assay and L-DOPA oxidation assay respectively. The cytokines such as ET-1 and SCF in HaCaT were detected by ELISA. Results A375 cells in co-culture system grew well, and 0.1, 0.5, 1.0, 5.0, 10 μg/mL Galangin of the purity of 70% and 90% had up-regulation effect on cell proliferation, melanin content and tyrosinase of A375. Moreover, Galangin could increase the expression level of ET-1 and SCF of HaCaT at more than 0.5 μg/mL, and the effect of Galangin of 70% purity was better than 90% purity. Conclusion Galangin could promote the cell proliferation, melanin content and tyrosinase of A375, and mechanism of the pathways is probably related to the up-regulation on the expression of ET-1 and SCF of HaCaT.