Objective To apply iTRAQ technology to observe changes in protein expression group in the process of inducing C2C12 cells differentiation towards osteoblast by BMP-2.Methods The myoblast C2C12 cells were seeded in BMP-2 induced differentiation system for differentiation induction.In the 7th day,differentiation protein was extracted and labeled with iTRAQ reagent.Then,mass spectrometric detection,data analysis of differentially expressed proteins,and analysis of biological information were carried out.Results 23 significantly differentially expressed protein spots were screened by BMP-2-induced myoblast C2C12 differentiated cell protein expression profile analysis,where the protein was labeled with iTRAQ reagent.8 protein points were up-regulated,and 15 protein points were down-regulated.Trend classification found that the above differential protein had differential expression in each period of C2C12 cell osteogenic differentiation (1-7 days).Part of up-regulated protein in the early differentiation period showed high expression level;part of up-regulated protein in the late differentiation period showed high expression level;similarly,part of down-regulated protein in the early differentiation period presented low expression level;part of down-regulated protein in the late differentiation period showed low expression level.Preliminary identification showed SERCA3,Cytochrome bS,S100A4,ATPase inhibitor and ATPIF1 presented dynamic changes,which suggests that these proteins may be related to inducing osteogenic differentiation mechanism.Conclusion The results of differential protein expression trend show the necessity of full monitoring of C2C 12 cells osteogenic differentiation and indicate that iTRAQ technology is an effective method of studying protein changes of cellular molecule.Five proteins including SERCA3,Cytochrome b5,S100A4,ATPase inhibitor and ATPIF1 can be used as candidate targets for osteogenic differentiation mechanism research.

OBJECTIVE：To investigate t he effects of different compatibility ratio of Gardenia jasminoides to fermented soybean on the content of genistein and total flavonoids ，and to investigate the compatibility regularity of Zhizichi decoction. METHODS：The decoction method was used to prepare the mixed decoction with different compatibility ratio of G. jasminoides to fermented soybean （2∶1，1∶1，1∶2，1∶4，m/m，the same hereinafter ）. UPLC-MS/MS method was used to determine the content of genistein in Zhizichi decoction with different compatibility ratio and corresponding fermented soybean single decoction. UV method was used to determine the content of total flavonoids in Zhizichi decoction with different compatibility ratio and corresponding gardenia single decoction and fermented soybean single decoction. RESULTS ：The established method had good linearity ， precision，repeatability，stability and accuracy. Compared with single decoction ，the content of genistein in the mixed decoction with different compatibility ratio of G. jasminoides to fermented soybean （2∶1，1∶1，1∶2，1∶4）was decreased to different extents ， while the content of total flavonoids was increased to different extents. With the increase of fermented soybean ，the content of genistein in the decoction increased at first and then decreased. When the compatibility ratios of G. jasminoides to fermented soybean were 1 ∶ 1 and 1 ∶ 2，the content of genistein in the decoction was the highest （all 0.071 μg/mL）. With the increase of fermented soybean ，the content of total flavonoids in the decoction did not change regularly ；when the ratio of G. jasminoides to fermented soybean was 1 ∶ 1，the content of total flavonoids in the decoction was the highest （1.861 μg/mL）. CONCLUSIONS ： When the compatibility ratio of G. jasminoides to fermented soybean was 1 ∶ 1，the content of flavonoids in the decoction is the highest.

OBJECTIVE To optimize th e p rocessing technology of Portulaca oleracea charcoal，and to investigate its improvement effect on the symptom of hemorrhoid model rats. METHODS The effects of roasting temperature ，dosage and roasting time on the processing technology of P. oleracea charcoal were investigated with Box-Behnken response surface methodology using comprehensive score of tannin content ，water-soluble extract content and appearance properties as the index. The optimal process parameters are selected and verified. The hemorrhoid model rats were treated with P. oleracea charcoal（0.8 g/mL）prepared by the optimal processing technology ，once a day ，for 11 days. After last medication ，the perianal pathological score of hemorrhoid model rats were performed ；serum levels of tumor necrosis factor α（TNF-α），interleukin 6（IL-6）and IL- 1β were detected. RESULTS The optimal processing technology of P. oleracea charcoal included roasting temperature of 200 ℃， dosage of 150 g and roasting time of 14 min. Results of validation test showed that the comprehensive score of P. oleracea charcoal was 92.57，and relative error of it with predicted value （96.59）was －4.13％. External use of P. oleracea charcoal 0.8 g/mL prepared by the optimal processing technology could significantly promote the wound healing of hemorrhoid model rats ，reduced the amount of exudate ，and decreased the levels of TNF-α，IL-6 and IL-β in serum. CONCLUSIONS The optimized processing technology of P. oleracea charcoal is feasible. P. oleracea charcoal prepared by the optimized processing technology has good curative effect on the symptom of hemorrhoid model rats.