| Gushudan prescription was formed according to "The Basic Conception of Establishing the Technical Operation Flat Stage for the Fined Selective System of Small Prescription of Chinese Herbs" and "The Database Management System of PUJIFANG". This prescription is composed of Herba Epimedii, Fructus Cnidii, Rhizoma Drynariae and Radix Salviae Miltiorrhizae, and commonly used for the treatment of osteoporosis.A high performance liquid chromatography (HPLC) method was developed for separation and determination of icariin, epimedin B and epimedin C in Herba epimedii. The contents of icariin, epimedin B, epimedin C in the species of epimedium from 16 sources were determined. The HPLC assay was performed on a Hypersil BDS C18 column using a mixture of acetonitrile - water (pH 3.0, 25: 75, v/v) as the mobile phase at a flow-rate of l.0mL min-1, a UV detector at 260 nm, and trimethoprim as internal standard. A good linearity between peak area ratio and the concentration was found in the range of 0.21~172 g mL-1 0.18 ~ 144 gmL-1 and 0.16~132 gmL-1 for icariin , epimedin B and epimedin C (r 0.999 6), respectively. The recoveries were 97.2%, 98.0% and 98.7%, with RSD of 2.0% 2.1% and 1.9% for icariin epimedin B and epimedin C, respectively (n = 9).Similarly, a method for determining othole and imperatorin in Fructus Cnidii was fulfilled by HPLC and the contents of othole and imperatorin in the species of Fructus Cnidii from 13 sources were determined. The chromatography was performed using a Kromasil Cg column with mobile phase of methanol-water (65 : 35, v/v). The monitoring wavelength was 310 nm. The flow rate was 0.8mLmin-1and fluocinonide served as an internal standard. A good linearity between peak area ratio and the concentration was found in the range of 13 ~ 208 gmL-1 for osthole (r > 0.999 8) and 8 ~ 128gmL-1 for imperatorin (r > 0.999 9), respectively. The average recoveries were 98.2% (RSD=1.7%, n=9) and 97.5% (RSD=2.0%, n=9) for osthole and imperatorin, respectively.An HPLC method for determining naringin and neoeriocitrin in Rhizoma Drynariae was also developed and the contents of naringin and neoeriocitrin in the species of Rhizoma Drynariae from 7 sources were determined. The HPLC assay was performed on a Hypersil ODS2 column using a mixture of acetonitrile - water (20: 80, v/v) as the mobile phase at a flow-rate of l.0mL min-1, and a UV detector at 283 run. A good linearity between peak area and the concentration was found in the range of 10 ~ 400 g mL-1 and 12 ~ 480 g mL-1 for naringin and neoeriocitrin (r 0.999 7), respectively. The recoveries were 100.0% and 99.4%, RSD were 1.4% and 2.0% for naringin and neoeriocitrin, respectively (n = 9).Osteoblasts are important cells in bone formation and, therefore, agents stimulating osteoblastic bone formation would have potential activity of anti-osteoporosis. In order to exploit novel and effective medicines preventing and treating osteoporosis and to promote the modernization of anti-osteoporostic Chinese Traditional Compound Medicines (CTCM), osteoblast-like UMR106 cells were employed as an in vitro model. The cell proliferation and differentiation were measured as active indices of stimulating osteoblastic bone formation. The orthogonal design method was used in the study of the disassemble formulae of Gushudan prescription. The effects of 16 subsidiary formulae from Gushudan prescription on osteoblastic proliferation were examined. The compatibility of the four botanical drugs in Gushudan prescription was demonstrated on cellular level and the rationality of prescription was proved in a modern scientific way. Gushudan prescription was examined extensively for the activity of stimulating osteoblastic bone formation in vitro in this study. The 75% ethanol extract of Gushudan prescription exhibited maximal activity promoting osteoblastic proliferation anddifferentiation. When the cells were cultured with 1.0 x l0-3mg mL-1 (expressed in the weight of raw materials) crude extract, an increase of 46.8% in osteoblastic proliferation; and an ALP...
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