AIM:To develop a capillary GC method for determining six components in Heiguidou Oil,?-pinene,eucalyptol,borneol,methyl salicylate,cinnamia aldehyde and eugenol. METHODS:GC was applied to quantitative analysis. Chromatographic conditions consisted of PEG-20M(30 m?0.25 mm?0.25 ?m),flame ionization detector(FID),and programmed temperature; initiation temperature was at 80 ℃ for 7 min; the rate of temperature was 15℃/minute,final temperature was at 180 ℃ for 10 min,bypass rate was 50 ∶1. The sample dissolved in absolute alcohol. The external standard was used. RESULTS:The standard curves of conclusion ?-pinene,eucalyptol,borneol,methyl salicylate,cinnamia aldehyde and eugenol were linear within the ranges of 0.322-32.150 mg/mL,0.202 -20.215 mg/mL,0.126-12.630 mg/mL,0.633-63.290 mg/mL,0.174-17.390 mg/mL,0.633-63.290 mg/mL and 0.203-20.255 mg/mL,respectively.RSD were 0.83%,0.48%,0.48%,0.31%,and 0.53 %,respectively. CONCLUSION:The established method is accurate and it can be applied to the simulta-neous assay of six components.

[ ABSTRACT] AIM:To investigate the effects of tanshinone IIA ( Tan IIA) on proliferation, apoptosis and its mo-lecular mechanism in human hepatoma HepG2 cells under hypoxic condition.METHODS:Hypoxia model was established by treatment with cobalt chloride ( CoCl2 ) .The cells were divided into normoxia control group, hypoxia control group and hypoxia combined at different concentrations of Tan IIA groups.After HepG2 cells were incubated with different concentra-tions of Tan IIA (0.5, 1.0, 2.0, 5.0 and 10.0 mg/L) for 24 h, 48 h and 72 h under hypoxic condition, the cell prolifer-ation was determined by MTT assay.After Tan IIA was added to the media at different concentrations for 24 h and 48 h, the apoptotic cells were observed by Hoechst 33258 staining.The protein levels of hypoxia-inducible factor 1 alpha (HIF-1α) , vascular endothelial growth factor ( VEGF) and wild-type P53 were detected by Western blotting after cultured with different concentrations of Tan IIA for 48 h.RESULTS:Tan IIA inhibited the proliferation of HepG2 cells in a dose-and time-dependent manner.Tan IIA induced the typical morphology of apoptotic cells and increased the apoptotic rate in a dose-and time-dependent manner after treatment with 1.0 mg/L~5.0 mg/L for 24 h and 48 h under hypoxic condition. The protein levels of HIF-1αand VEGF were weakly expressed in HepG2 cells under normoxia but up-regulated after incu-bated under hypoxia for 48 h.The protein expression of HIF-1αand VEGF were decreased with the increase in the concen-tration of Tan IIA under hypoxia.The protein expression of wild-type P53 was increased with the increase in the concentra-tions of Tan IIA under hypoxia.CONCLUSION:Tan IIA significantly inhibits the proliferation and induces the apoptosis of human hepatoma HepG2 cells under hypoxia, which may be related to the down-regulation of HIF-1αand VEGF and up-regulation of wild-type P53.

Aim To investigate the mechanism of demethylation on adenosine (ADO )and homocysteine (HCY)-induced apoptosis in human hepatoma HepG2 cells .Methods HepG 2 cells were treated with differ-ent concentrations of ADO (1.0、2.0、4.0 mol · L-1 ) alone or in combination with HCY for 6h,12h and 24h,5-aza-2-deoxycytidine (5-Aza-CdR)as a positive control.Cell viabilities were assessed by CCK8 assay. Cell apoptosis was observed by AnnexinV-FITC/PI staining.The mitochondrial membrane potentials(ΔΨ) were measured by flow cytometry.The mRNA and pro-tein expressions of caspase-3,caspase-8,caspase-9, MDM-2,p53,Cytochrome C,DNMT1,DNMT3a,DN-MT3 b were detected by RT-qPCR and Western blot re-spectively.Results ADO alone or in combination with HCY significantly reduced the viability of HepG2 cells in a dose and time-dependent manner.The apoptotic rates of HepG2 cells after combination treatment with ADO and HCY at 1 .0,2.0,4.0 mol · L-1 for 24 h were (1 8.63 ± 1.25 )%,(29.42 ±2.37 )% and (42.47 ±3.09 )%,compared with the control group (1.30 ±0.82 )%,P <0.01;and the mitochondrial membrane potentials were decreased from 674.15 ± 82.8%(black control group)to (428.38 ±54.5)%, (297.78 ±30.5)%,(74.45 ±5.73)%,P<0.01, respectively.The expressions of caspase-3,caspase-8, caspase-9,MDM-2,p53,Cytochrome C were up-regula-ted and MDM-2 were down-regulated after combination treatment of ADO and HCY.The mRNA expressions of DNMT1 ,DNMT3 a and DNMT3 b were down-regulated after combination treatment with ADO and HCY or 5-Aza-CdR alone.Conclusion Combination treatment of ADO and HCY can cause cellular methylation chan-ges.The effects of demethylation of ADO and HCY may activate p53 gene and mitochondrial pathway, which at last leads to HepG2 cell apoptosis.

Objective To evaluate the efficacy of Budesonide/formoterol to control asthma under real-life conditions.Methods A muhi-center, open label, non-interventional study was conducted.Asthma control after 12 week therapy with Budesonide/formoterol was assessed by Asthma Control Questionnaire (ACQ) and modified Asthma Control Questionnaire (ACQ5).Results A total of 360 asthma patients were recruited,including 228 adult patients and 132 child patients.After 12 weeks' therapy,all the patients' medium value of ACQ was decreased significantly from 2.03 (adults 2.20, children 1.74) at baseline to 0.60 (adults 0.78, children 0.29) (P < 0.0001), and the medium value of ACQ5 was also decreased significantly from 2.4 (adults 2.24, children 1.76) at baseline to 0.47 (adults 0.62, children 0.20) (P < 0.0001).Conclusion Budesonide/formoterol is effective in asthma treatment, by which most asthma patients obtain and maintain clineal control.