Objective:To study the alkaloids in Macleaya cordata. Methods:The compounds were isolated and purified by silica gel,recrystallization,and semi-preparative HPLC, and their strcuctures were confirmed by MS and NMR. Results: Nine alkaloids were obtained and identified as dihydrochelerythrine(1), 6-cyanodihydrosanguinarine(2), R-6-((R)-1-hydroxyethyl)dihydrochel-erythrine(3),8-demethyl-dihydrochelerythrine(4),cheilanthifoline(5),canadine(6),8-demethyl-chelerythrine(7),protopine(8) and allocryptopine(9). Conclusion:Compounds 4-6 are isolated from the genus of Macleaya for the first time.

Objective:To investigate the absorption characteristics of matrine and glycyrrhizic acid in different intestinal segment and at various mass concentration,and explore the absorption changes of the combination of the two drugs.Methods:In situ single pass intestinal perfusion (SPIP) model was used,and the concentration changes of matrine and glycyrrhizic acid were determined by HPLC, and Kaand Peffwere calculated. Results:Matrine was absorbed in the whole of small intestine, and best in jejunum, and compared with the duodenum and ileum,the absorption had significant difference (P<0.05).The absorption at different mass concentrations had significant difference(P<0.05). The absorption of glycyrrhizic acid in small intestine was poor without significant differences in different intestinal segments and at various doses (P>0.05). After the combination of the two drugs, the absorption of matrine and glycyrrhizic acid had significant difference when compared with that of separated use(P<0.05). Conclusion:The absorption of ma-trine is dose-dependent,and with the dose increase,the absorption increases. Glycyrrhizic acid absorption does not change with mass concentration,and the absorption may be passive transport. The combination of the two drugs can improve the in vivo absorption of ma-trine and increase the concentration of glycyrrhizic acid in intestine.

Objective:To study the chemical constituents in the roots of Tripterygium wilfordii. Methods: The compounds from Tripterygium wilfordii were isolated and purified by silica gel, Sephadex LH-20 and prep-HPLC chromatography, and their structures were elucidated based on the physiochemical properties and spectroscopic analysis. Results:Twelve compounds were isolated and iden-tified as wilforgine(1),wilforine(2),triptonoterpene methyl ether(3),glut-5-en-3β,28-diol(4),wilforol E(5),triptobenzene L (6),maytenoic acid(7),triptophenolide(8),celastrol(9),demethylzeylasteral(10),1-desacetyl wilforgine(11) and wilfortrine (12). Conclusion:The 1D and 2D NMR data of 1 and 2 are assigned for the first time,and the absolute configurations of 1 are con-firmed by X-ray single crystal diffraction.

Objective:To establish a method for the determination of 11 trace elements in white vaselin. Methods:The contents of Mg,Al,Fe,Co,Ni,Cu,Zn,As,Cd,Hg and Pb in white vaselin were determined by inductively coupled plasma mass spectrome-try (ICP-MS). The parameters of ICP-MS were as follows:the Argon gas pressure was 0.6 MPa,the helium pressure was 0.12 MPa, the scan times were 200,the flush time was 45 s,the sampling time was 45 s and the internal standard was added online. Results:The linear relationship between concentration and response value of each element standard solution was within the range of 0-100 ng· m L-1. Except the correlation coefficients of Mg and Fe were 0.998 and 0.997,that of the other elements was all above 0.999. The recoveries of Mg,Fe,Ni,Zn,As and Pb were 74.9%-83.0%,those of Cd and Hg were 87.5%-94.4%,and those of Al,Co and Cu were 107.6%-118.7%. The detection limit of Mg, Al, Fe, Co, Ni, Cu, Zn, As, Cd, Hg and Pb was 1.9,0.59,0.61,0.16, 0.33,1.5,1.7,0.09,0.12,0.70 and 1.6 ng·ml-1,respectively. Among the 67 samples,the contents of Mg,Al,Fe,Ni,Cu, Zn, As and Hg were all detected out,while those of Co,Cd and Pb were only detected out in some samples. Conclusion:The method has the advantages of simple operation and high sensitivity,which can satisfy the determination of trace elements. It is suggested to se-lect 4 elements (Mg,Al,Fe and Ni) as the evaluation indices for white vaselin.

Objective:To establish an HPLC method for the determination of paraquat in human serum.Methods:The analytical column was a Kromasil C18column (200mm×4.6mm,5μm).The mobile phase was a mixture of acetonitrile and water ( containing 0.03 mol·L-1sodium heptanesulfonate and 0.24 mol·L-1phosphoric acid) (3 :97,pH was adjusted to 2.0 by triethylamine),the detection wavelength was set at 258 nm,the column temperature was 25℃,the injection volume was 20 μl,and the flow rate was 0.8 ml·min-1.Results:The calibration curve of paraquat was linear within the range of 0.106-10.6 mg·L-1( r =0.999 3),and the lower limit of detection was 0.065 mg·L-1. The absolute recovery of paraquat at low,medium and high concentration was more than 89.4%,and the method recovery was more than 94.4%. The intra-day RSDs were 0.12%-1.74%,and the inter-day RSDs were 0.44%-2.89%.Conclusion:The method is simple,quick,accurate,sensitive and specific,and can be used for detecting paraquat con-centration in human serum.

Objective:To investigate the effects of metformin on myocardial cell apoptosis during myocardial ischemia-reperfusion (MI/R) injury in rats. Methods:The rat model of MI/R injury was prepared by coronary artery ligation for 30 min followed by 2-hour reperfusion. Then the rats were randomly divided into 6 groups: the normal control (NC) group, the sham group, the model (IR) group,metformin group respectively at low (LD),medium(MD) and high(HD) dose(150,300 and 500 mg·kg-1) with 10 ones in each. From 3 days before the model establishment to death,the rats were with gavage administration every day. The myocardial cell apoptosis was detected by the TUNEL method. The expression of HIF-1α gene in myocardial tissues was detected by the RT-PCR meth-od. The expressions of Fas,Bcl-2,Bax and caspase-3 proteins in myocardial tissues were tested by Western blot. Results:Compared with that in the IR group,the myocardial cell apoptosis rates in LD group,MD group and HD group all decreased,and the differences were statistically significant(P<0.01),meanwhile,the relative expression levels of HIF-1α mRNA in myocardial tissues at 24 h and on the 7th day in LD group,MD group and HD decreased,and the differences were statistically significant(P<0.01),and the rela-tive expression levels of Fas,Bax and caspase-3 proteins at 24 h and on the 7th day in LD group,MD group and HD group decreased, while the relative expression levels of Bcl-2 proteins increased,and the difference were statistically significant(P<0.01). The effect of metformin in each dose group was dose-dependent,and the differences were statistically significant(P<0.01) when compared MD group and HD group with LD group. Conclusion:Metformin can reduce myocardial cell apoptosis during myocardial ischemia-reperfu-sion injury in rats. The mechanism may be related to the improvement of myocardial ischemia environment,inhibition of Fas death sig-naling pathway activation and increase apoptosis inhibitor Bcl-2 expression.

Objective:To explore the correlation between the environmental factors and primary metabolite of Taxus mairei (Le-mee et Levl.) S.Y. Hu ex Liu. and to provide experimental basis for the further discussion on the biological characteristics,develop-ment and utilization of Taxus mairei (Lemee et Levl.) S.Y. Hu ex Liu.. Methods:Correlation analysis and path analysis were used to explore the correlation of temperature, sunshine duration and precipitation with the relative primary metabolite content by SPSS 19.0. Results:The environmental factor with the biggest direct contribution to the absolute value of relative primary metabolite content was average monthly temperature. The average monthly temperature by month cumulative sunshine time showed the largest influence on the relative primary metabolite content. Conclusion:The relationship between temperature and sunshine time is much close, and the most obvious factor on the primary metabolite accumulation is temperature changes. This research provides theoretical basis and certain guiding significance for the further development,utilization and studies on harvesting,processing, using and explaining the biological characteristics of Taxus mairei (Lemee et Levl.) S.Y. Hu ex Liu..

Objective:To observe the effect of intrathecal administration of 8-O-acetyl-SM(8-OaS) on chronic neuropathic pain in rats by inhibiting the expression of protein kinase B(AKT)-mammalian target of rapamycin(mTOR) signaling pathway in spinal dorsal horn after spinal nerve ligation.Methods:The rat model of neuropathic pain was established by lumbar 5 spinal nerve ligation(SNL), and Von Frey filament was used to investigate the mechanical allodynia. Immunofluorescent histochemistry was adopted to investigate the distribution of pAKT and pmTOR in spinal dorsal horn. The protein levels of pAKT and pmTOR in spinal dorsal horn after drug ad-ministration were quantitatively determined by Western blot. Results: Compared with that in the sham group, the paw withdrawal threshold (PWT) significantly decreased(P<0.01),and the intrathecal administration of 8-OaS attenuated mechanical allodynia ob-viously during the first day and the seventh day after the operation(P<0.05). Meanwhile,double immunofluorescent staining showed the co-expression of pAKT and astrocytes marker glial fibrillary acidic protein(GFAP),and positive labeling of pmTOR was expressed in spinal astrocytes and neurons. The results of Western blot revealed that the protein levels of pAKT and pmTOR in spinal dorsal horn were significantly reduced after the treatment of 8-OaS. Conclusion:Intrathecal administration of 8-OaS attenuates the PWT of SNL-in-duced chronic neuropathic pain. The underlying mechanism of the potential anti-allodynia effect of 8-OaS may be related to the suppres-sion of spinal astrocytes via decreasing the phosphorylation of AKT-mTOR signaling pathway resulting in attenuating the development of neuropathic pain.

Objective:To develop an HPLC-MS determination method for the effects of Xuesaitong on the pharmacokinetics of losartan in rats. Methods:After oral administration of single losartan and combined usage of losartan and Xusaitong in rats,the con-centrations of losartan and EXP3174 in plasma were determined by HPLC-MS. The main pharmacokinetic parameters were calculated and compared by statistical analysis. Results:The calibration curves of losartan and EXP3174 were linear within the range of 2.0-2 000 ng·ml-1(r=0.996 4 and r=0.999 5). The average recovery was 85.19%-109.79%. The results showed that after oral co-ad-ministration with Xuesaitong,t1/2and Vz/F of losartan significantly decreased (P<0.05), Tmaxsignificantly increased (P<0.05), t1/2of EXP3174 significantly decreased(P<0.05),and Tmaxsignificantly increased(P<0.05). Conclusion:The HPLC-MS method is sensitive and selective,and suitable for the detection of losartan and EXP3174 in plasma in vivo. The experiments showed that Xue-saitong can affect the absorption and metabolism of losartan in vivo.

Objective:To study the effects of voriconazole sulfonated butyl ether-β-cyclodextrin inclusion compound(VCZ-SBE) eye drop on fungal keratitis (FK) in rabbits. Methods:Aspergillus fumigatus was used for the establishment of FK model in rabbits. Totally 48 healthy rabbits were divided into four groups. The 12 right eyes in group A were inoculated with Aspergillus fumigatus with-out any treatment. The 12 right eyes in group B were inoculated with Aspergillus fumigatus,and then treated with 0.5% VCZ-SBE eye drop,those in group C were inoculated with Aspergillus fumigatus,and then treated with 1.0% VCZ-SBE eye drop,and those in group D were inoculated with Aspergillus fumigatus,and then treated with 0.5% fluconazole eye drop. The cornea of normal left eyes in the four groups was used for the blank control. Ulcer grading was performed for all the eyes in 0,1,4,7 and 14 d after the successful es-tablishment of FK model. Every 3 rabbits were sacrificed in 1,4,7 and 14 d,and then the cornea was obtained to evaluate the infiltra-tion degree of inflammatory cells and the invasion depth of hyphae into cornea. Results:After the successful establishment of FK mod-el,the ulcer grading among the groups showed no significant difference(P>0.05). Respectively after the 7,4 and 14-day treatment, the ulcer grading in group B,C and D was notably lower than that in group A (P<0.05). After the 4,7 and 14-day treatment,the ulcer grading,infiltration depth and necrosis degree in group C were all lower than those in group B and D(P<0.05),and after the 7 and 14-day treatment,those in group B were lower than those in group D(P<0.05). Conclusion:Compared with fluconazole eye drop at the same concentration,VCZ-SBE inclusion compound eye drop can decrease the ulcer grading of FK and reduce the infiltration depth in a concentration-dependent manner.