AIM: To establish the determination of narigin and hesperidin in Juhong Pills(Exocarpium, Citri Grandis, Pericarpium Citri Reticulatae, Radix Angelicae Sinensis, etc.). METHODS: The extraction was completed with ether. The Shim-pack ODS(?6.0mm?150mm) column was used with mobile phase of CH 3OH-CH 3COOH-H 2O(30∶4∶60). The detection wavelength was at 283nm. RESULTS: The linear range for narigin was 0.072～1.43?g, r=0.9999 and the linear range for hesperidin was 0.068～1.37?g, r=0.9999, respectively. Both the average recoveries were 99.3% and 99.4%, respectively. Both RSD were 0.6%(n=5). CONCLUSION: The method is simple and the result is reliable.

This study was aimed to optimize the extraction process of ginseng in Shen-Qi Bu-Qi (SQBQ) granules. The orthogonal experiment method was used to optimize the extraction process of ginseng adopting ethanol concentration, ethanol volume, extraction time and extraction times as factors. The content of ginsenoside Rg1, Re, Rb1 content in the extract was determined by HPLC. And the optimum extraction conditions were determined with indexes of three kinds of saponin yield. The results showed that the optimum extraction process of ginseng for SQBZ granules was that adding 6 times amount of 60% ethanol technology for medicinal materials, extract for 2 times, and 3 h for each time. It was concluded that the optimization of ginseng extraction process was stable, reasonable and feasible with high extraction rate.

Objective:To investigate the effect of mechano-growth factor（MGF）on osteoclast activity and its mechanism.Methods:The RAW264.7 precursor osteoclast cell line was cultured with 25 ng/ml macrophage-colony stimulating factor（M-CSF）and 30 ng/ml receptor activator of NF-κB ligand（RANKL），and identified by tartrate resistant acid phosphatase（TRAP）staining after 7 days of culture. Western blot anslysis was used to determine the effect of 45 ng/ml MGF on the phosphoinositide-3-kinase/protein kinase B（PI3K/AKT）signaling pathway in separated osteoclasts，including levels of AKT，phosphorylation（p）-AKT，lactation mammalian target of rapamycin（mTOR），p-mTOR and TRAP at 0，4，8 and 12 hours. Real-time fluorescence quantitative PCR was used to expressions of TRAP in osteoclasts at 0，4，8 and 12 hours. The PI3K/Akt phosphorylation inhibitor LY294002（20 μmol/L）combined with MGF（45 ng/ml）was used to act on osteoclasts，and expression levels of Akt，p-Akt，mTOR，p-mTOR and TRAP were detected by Western blot at 0，4，8 and 12 hours.Results:After culturing RAW264.7 cells with M-CSF and RANKL for 7 days，a large number of osteoclasts with positive TRAP staining can be obtained. Western blot analysis showed expression levels of Akt and mTOR did not change significantly over time（ P>0.05），expression levels of p-Akt and p-mTOR increased continuously from（2.18±0.34）pg/ml and（0.83±0.10）pg/ml at 0 hour to（3.86±0.36）pg/ml and（1.56±0.19）pg/ml at 12 hours（ P<0.05），and expression level of TRAP decreased significantly over time，from（5.66±0.47）pg/ml at 0 hour to（3.76±0.38）pg/ml at 12 hours（ P<0.05）. Real-time fluorescence quantitative PCR analysis of expression of TRAP in osteoclasts showed that MGF inhibited the expression of TRAP in osteoclasts，which decreased from 1.02±0.06 at 0 hour to 0.53±0.11 at 12 hours（ P<0.05）. After acting LY294002 combined with MGF on osteoclasts，Western blot analysis showed expression levels of Akt and mTOR did not change significantly over time（ P>0.05），expression levels of p-AKT and p-mTOR decreased significantly from（3.28±0.18）pg/ml and（3.29±0.22）pg/ml at 0 hour to（2.06±0.34）pg/ml and（2.04±0.20）pg/ml at 12 hours（ P<0.05），and expression level of TRAP had no significant difference over time（ P>0.05）. Conclusions:MGF inhibits osteoclast activity by inhibiting the expression of TRAP in osteoclasts through PI3K/Akt signaling pathway. LY294002 inhibits the expression of PI3K/Akt signaling pathway in osteoclasts，further verifying the mechanism of MGF inhibiting osteoclast activity，and this finding puts forward new ideas for clinical prevention and treatment of osteoporosis.