Objective To develop a RP-HPLC method for the determination of Brucine and Strychnine in Semen Strychni and its extractive of total alkaloids. Methods A chromatographic column of Licrospher C18 (4.6 mm×250 mm, 5 μm) was used with the mobile phase of acetonitrile∶0.01 mol/L sodium heptane sulfonate and 0.02 mol/L potassium dihydrogen phosphate mixed with equal amount (adjusted pH to 2.8 with 10% phosphonic acid)=27∶73, detection wavelength at 260 nm, column temperature of 30 ℃ and flow rate of 1 mL/min. Results The calibration curves of Brucine and Strychnine were both in good linearity in the ranges of 0.1-1.0 μg and 0.12-1.2 μg (r=1.000) respectively. The average recovery rates of Brucine and Strychnine were 99.88% (RSD=1.06%) and 100.06% (RSD=0.78%) respectively. Conclusion The method is realiable and accurate, which can be applied to determination of Brucine and Strychnine in Semen Strychi and its extractive.

BACKGROUND:Osteogenesis is closely integrated with angiogenesis in bone formation and repair process, and hypoxia-inducible factor 1 alpha (HIF-1α) is considered to be the most important core transcription factor promoting angiogenesis gene regulation, which may promote the formation of blood vessels at hypoxia portion, and thus contribute to bone formation.
OBJECTIVE:To construct the Lenti-HIF-1α-IRES-EGFP (wild type) and Lenti-HIF-1α-IRES-EGFP (point mutant type) lentiviral eukaryotic expression vectors and to detect their impact on the osteogenic gene expression of rabbit bone marrow mesenchymal stem cells.
METHODS:The Lenti-HIF-1α-IRES-EGFP (wild type) and Lenti-HIF-1α-IRES-EGFP (point mutant type) lentiviral eukaryotic expression vectors were constructed according to the wild type human HIF-1αgene sequence and the determined restriction sites of human HIF-1αpoint mutant sequence. Rabbit bone marrow mesenchymal stem cells were transfected with the prepared Lenti-HIF-1α-IRES-EGFP (wild type) and Lenti-HIF-1α-IRES-EGFP (point mutant type) virus solution.
RESULTS AND CONCLUSION:Immunofluorescence microscopy observations indicated that the cells of Lenti-HIF-1α-IRES-EGFP (wild type) group and Lenti-HIF-1α-IRES-EGFP (point mutant type) group showed no obvious fluorescence on transfected 7 days, and two groups of cells showed a more obvious green fluorescent after transfection 14 days. Quantitative PCR analysis results showed that there were obvious HIF-1αand bonemorphogenetic protein 2 gene expressions on days 7 after transfection and the two genes stil showed highly expression levels after transfection 14 days. The two lentiviral eukaryotic expression vectors of Lenti-HIF-1α-IRES-EGFP (wild type) and Lenti-HIF-1α-IRES-EGFP (point mutant type) could be constructed according to the wild type human HIF-1αgene sequence and the determined restriction sites of human HIF-1αpoint mutant sequence;HIF-1αgene can promote the osteogenic gene expression of lentivirus-transfected rabbit bone marrow mesenchymal stem cells.

Objective:To investigate the effects of icaritin(ICT)on the proliferation and osteogenic differentiation of rat bone mar-row stromal cells(rBMSCs).Methods:rBMSCs were cultured from the bone marrow of SD rats and identified by multilineage differ-entiation assays.3,6 and 9 days after the treatment of rBMSCs of passage 4 by ICT at 1 0 -9 ,1 0 -8 ,1 0 -7 ,1 0 -6 and 1 0 -5 mol/L re-spectively,the proliferation and differentiation of the cells were examined by cck-8 and alkaline phosphatase (ALP)activity assay kit respectively.The calcium nodule formation was observed by alizarin red(AR)staining 21 days after 1 0 -9 mol/L ICT treatment. Results:Primary rBMSCs showed the typical spindle-like shape with attachment growth.rBMSCs could be induced to osteogenic and adipogenic differentiation.The proliferation of rBMSCs was inhibited but ALP activity was enhanced by ICT.1 0 -9 mol/L ICT in-cresed calcium nodule formation.Conclusion:ICT can dose-dependently inhibit the proliferation,but promote the osteogenic differ-entiation of rBMSCs.

Objective To observe the clinical effect of monosialotetrahexosyl ganglioside sodium injection (GM1) on spastic cerebral palsy. Methods 98 children with spastic cerebral palsy were randomly divided into control group (n=50) and treatment group (n=48). Both groups received Bobath approach, and the treatment group received GM1 in addition. They were assessed with Functional Independence Measure for Children (WeeFIM), Gross Motor Function Measure (GMFM) and Gesell Development Schedule (GDS) before and after 90 days of treatment. Results The scores of WeeFIM, all the dimensions of GMFM and the gross motor, fine motor, personal-social and adaption of the GDS improved in both groups after treatment (P<0.05), and improved more in the treatment group than in the control group (P< 0.05). Conclusion GM1 may further improve the recovery of function for children with spastic cerebral palsy.

Objective To explore the clinical efficacy and drug-toxic reactions of raltitrexed combined with oxaliplatin (RALOX protocol) and 5-fluorouracil + calciumfolinate + oxaliplatin (FOLFOX 4 protocol) in the treatment of patients with middle and advanced primary liver cancer (PLC).Methods A total of 72 patients with PLC were selected and randomly divided into RALOX group (n =34) and FOLFOX 4 group (n =38).The objective response rate (RR) was evaluated every 6 weeks after chemotherapy,while objective remission rate (OR),disease-control rate (DCR),median survival rate (mOS),median progression-free survival (mPFS),1-year survival rate (SR) as well as toxic and adverse reactions were observed.Results In RALOX group,31 patients were evaluable,with OR,DCR,mOS,mPFS,and 1-year SR being 19.4％,51.6％,7.2 months,3.4 months,and 22.6％,respectively.In FOLFOX 4 group,29 patients were evaluable,with OR,DCR,mOS,mPFS,and 1-year SR being 13.8％,48.3％,6.9 months,3.3 months and 20.7％,respectively.RALOX group was significantly lower than FOLFOX 4 group in the incidence rates of gastrointestinal reactions,liver toxicity,cardiac toxicity,peripheral nervous toxicity and hand-foot syndrome,but there were no significant differences in the incidence rates of renal toxicity and myelosuppression between two groups.Conclusion RALOX is safe and effective in the treatment of patients with middle and advanced PLC,and is superior to FOLFOX 4 protocol in clinical efficacy with mild adverse reactions.

Objective To explore the clinical efficacy and drug-toxic reactions of raltitrexed combined with oxaliplatin (RALOX protocol) and 5-fluorouracil + calciumfolinate + oxaliplatin (FOLFOX 4 protocol) in the treatment of patients with middle and advanced primary liver cancer (PLC).Methods A total of 72 patients with PLC were selected and randomly divided into RALOX group (n =34) and FOLFOX 4 group (n =38).The objective response rate (RR) was evaluated every 6 weeks after chemotherapy,while objective remission rate (OR),disease-control rate (DCR),median survival rate (mOS),median progression-free survival (mPFS),1-year survival rate (SR) as well as toxic and adverse reactions were observed.Results In RALOX group,31 patients were evaluable,with OR,DCR,mOS,mPFS,and 1-year SR being 19.4％,51.6％,7.2 months,3.4 months,and 22.6％,respectively.In FOLFOX 4 group,29 patients were evaluable,with OR,DCR,mOS,mPFS,and 1-year SR being 13.8％,48.3％,6.9 months,3.3 months and 20.7％,respectively.RALOX group was significantly lower than FOLFOX 4 group in the incidence rates of gastrointestinal reactions,liver toxicity,cardiac toxicity,peripheral nervous toxicity and hand-foot syndrome,but there were no significant differences in the incidence rates of renal toxicity and myelosuppression between two groups.Conclusion RALOX is safe and effective in the treatment of patients with middle and advanced PLC,and is superior to FOLFOX 4 protocol in clinical efficacy with mild adverse reactions.