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OBJECTIVE:To estab lish a method that can comprehensively and rapidly analyze the chemical compositions of Miao medicine Caesalpinia decapetala,and to providing reference for quality control and pharmacodynamic material basis study of C. decapetala . METHODS :UPLC-Q-TOF-MS/MS was adopted . The determination was performed on Agilent SB-C 18 column with mobile phase consisted of 0.1% formic acid solution- 0.1% formic acid acetonitrile (gradient elution )at the flow rate of 0.25 mL/min. The column temperature was 30 ℃,and sample size was 2 µL. ESI source was applied in negative and positive scanning ion mode and data collection range of m/z 50-1 500. The capillary voltage was 4.5 kV,the atomizing gas (nitrogen)pressure was 1.2 Bar, the solvent removal gas was nitrogen ,the flow rate of solvent removal gas was 8 L/min and the solvent removal gas temperature was 200 ℃. Data Analysis 4.2 software was adopted to analyze fragment ion information of each peak ,and identify chemica l compositions on the basis of relevant literature and mass spectograms of reference substance. RESULTS :Under positive ion mode , 9 chemical compounds were identified ;peak 1,2,3,4,5,6,7,8,9 were catechin ,protohematoxylin B ,epicatechin,ethyl gallate,quercetin,luteolin,3-deoxy-hematoxylin chalcone , isoliquiritigenin and linoleic acid. Under negative ion mode , U1812403), totally 21 peaks were confirmed and 13 compounds were identified;peak 3,4,5,6,7,8,9,10,11,12,13,15, 21 were catechins , brevifolin carboxylic acid , proto- hematoxylin B ,epicatechin,ethyl gallate ,epicatechin gallate , quercetin,resveratrol,hematoxylin,luteolin,3-deoxy-hema- toxylin, isoliquiritigenin, linoleic acid. CONCLUSIONS UPLC-Q-TOF-MS/MS method is established successfully for analysis of chemical compositions in C. decapetala .
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OBJECTIVE: To study in vitro metabolism pathway of effective component of Bletilla striata as Militarine in liver microsomes and kinetics characteristics of enzyme-catalyzed reactions. METHODS: The in vitro incubation system of rat and human liver microsomes was established, and incubation reaction of Militarine was performed. UPLC-QTOF-MS was used to identify the structure of its metabolites in combination with UNIFI database and references. Using puerarin as internal standard, UPLC-Triple Quad-MS was used to quantitatively analyze metabolic transformation of Militarine in rat liver microsomes. The kinetic parameters (vmax, km, CLint) of Militarine enzyme-catalyzed reactions with/without reducing coenzyme Ⅱ (NADPH) were calculated by fitting the curves with GraphPad Prism 5.0 software. RESULTS: After incubation in rat and human liver microsomes, Militarine produced a chemical formula C21H29O11, which was presumed to be a metabolite of Militarine ester bond hydrolysis. The kinetic study of enzyme-catalyzed reactions showed that vmax of Militarine enzyme-catalyzed reactions with/without NADPH were 1.955, 2.129 nmol/(h·mg); km were 8.601, 9.854 nmol/mL; CLint were 0.227 3, 0.216 1 mL/(h·mg); there was no significant difference between with NADPH and without NADPH. CONCLUSIONS: The main metabolic pathway of Militarine in liver microsomes is the hydrolysis of C1 and C4 ester bonds. Its metabolism does not depend on the pathway of cytochrome P450 enzymes initiated by NADPH.
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OBJECTIVE: To study the protective effects of Polygonum orientale extract on myocardial ischemia-reperfusion injury (MIRI) model rats, and to provide reference for it’s deeply development of medicinal source. METHODS: Totally 24 rats were randomly divided into sham operation group (normal saline), model group (normal saline), Compound danshen tablet group (positive group, 0.17 g/kg) and P. orientale extract group (86 g/kg, calculated by crude drug), with 6 rats in each group. All groups were given drugs 2 mL/100 g intragastrically once a day. After 4 d of consecutive administration, MIRI model was induced by the left anterior descending branch of arteria coronaria in all groups except for sham operation group. 24 h after reperfusion, they were given related medicine again. After medication, the changes of electrocardiogram ST segment were monitored in each group. The plasma levels of LDH, CK-MB, CK, cTn-I, SOD and NO were detected in each group. The myocardial infarction rate in each group was calculated and the pathomorphological changes in the myocardium were observed. RESULTS: Compared with sham operation group, ST segment of myocardial electrocardiogram was increased in model group (P<0.01). The plasma levels of LDH, CK, CK-MB and cTn-I were increased significantly (P<0.01), while the plasma levels of SOD and NO were decreased significantly (P<0.01). The rate of myocardial infarction was increased significantly (P<0.01), and pathomorphological changes were observed in myocardial tissue such as infiltration of inflammatory cells and loose cytoplasm of cardiac myocytes. Compared with model group, ST segment of myocardial electrocardiogram was decreased significantly in Compound danshen tablet group and P. orientale extract group (P<0.05); the plasma levels of LDH, CK, CK-MB and cTn-I were decreased significantly (P<0.05), while the plasma levels of SOD and NO were increased significantly (P<0.05); the rate of myocardial infarction was decreased significantly (P<0.05), and inflammatory cell infiltration and tissue edema in myocardium were relieved to varying degrees. CONCLUSIONS: The protective effect of P. orientale extract protect on MIRI may be exerted by anti-oxidative damage.
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Objective To evaluate the value of diagnostic negative MDCT cholangiopancreatography in patients with suspected obstructive biliary diseases by receiver operating characteristic(ROC)curve. Methods Dual-phases contrast-enhanced MDCT of the abdomen was performed in 30 patients.Multiplanar reformation,minimum intensity projection and volume rendering were generated using original data in vein phase.By double-blind method,the images were readed by two radiologists.ROC curves were analysed with software SPSS11.5,the different results of reading images between two radiologists was evaluated with Kappa test.Results The Az of the area under the ROC curve of MDCT cholangiopancreatography for the two observers was 0.968 and 0.962 respectively,it was more than 0.9,which showed that the value of diagnosing the suspected obstructive biliary diseases by negative MDCT cholangiopancreatography was good.Kappa index test was satisfactory.Conclusion Negative MDCT cholangiopancreatography may be a routine choice in diagnosing the suspected obstructive biliary diseases.