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OBJECTIVE: To study the protective effects of Pericarpium Trichosanthis extract(TPE) on H9c2 myocardial cells injured by hypoxia/reoxygenation(H/R). METHODS: H/R injury model of H9c2 myocardial cells was established by using sodium dithionite (Na2S2O4) as chemical hypoxia agent. The modeling conditions of different concentrations of Na2S2O4 (0.625-10 mmol/L) and different time of hypoxia (10-60 min) and reoxygenation (2-8 h), as well as the concentration of TPE (12.5-400 μg/mL) were screened. The cultured H9c2 myocardial cells were randomly divided into normal group, model group, TPE different dose groups (TPE low-dose, medium-dose and high-dose groups, 25, 50, 100 μg/mL) and positive control group (quercetin, 25 μmol/L). They were pre-treated with relevant medicine for 24 h, and then H/R injury model was established. Cell viability was measured by MTT assay. The levels of LDH, CK-MB, SOD and MDA in cells were tested by ELISA. Apoptosis of H9c2 myocardial cells were observed by flow cytometry. Western blotting was used to detect the protein expression of Bax and Bcl-2 in cells. RESULTS: The condition of H/R injury modeling included modeling concentration of Na2S2O4 2.5 mmol/L, hypoxia time 30 min, reoxygenation time 4 h. 12.5-400 μg/mL TPE showed no toxicity to H9c2 myocardial cells. After treatment, compared with blank group, survival rate and apoptotic rate of H9c2 myocardial cells in model group were increased significantly; the levels of CK-MB, LDH and MDA were increased significantly, while SOD level was decreased significantly; protein expression of Bax and Bax/Bcl-2 ratio were increased significantly, while that of Bcl-2 was decreased significantly (P<0.05 or P<0.01). Compared with model group, above changes of H9c2 myocardial cells were reversed in all dose groups of TPE (P<0.05 or P<0.01). CONCLUSIONS: TPE can protect H9c2 myocardial cells against H/R injury. Its mechanism may be associated with inhibiting the increase of lipid peroxide, improving the ability of scavenging reactive oxygen free radicals, up-regulating the protein expression of Bcl-2 or down-regulating protein expression of Bax, so as to inhibit the cell apoptosis.
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OBJECTIVE:To establish a method for the content determination of total flavonoids in Morus alba. METHODS:UV-visible spectrophometry was performed with Al(NO2)3-NaNO2-NaOH color-test at the wavelength of 510 nm with the reference of rutin. RESULTS:The linear range of rutin was 0.031 2-0.156 mg/ml(r=0.999 9);RSDs of precision,stability and reproduc-ibility tests were lower than 2%;recovery was 95.7%-101.0%(RSD=2.1%,n=6). CONCLUSIONS:The method is simple,sta-ble and reproducible,and can be used for the content determination of total flavonoids in M. alba.
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OBJECTIVE:To explore the transform of the medicinal properties of Coptis chinensis-Cinnamomum cassia couplet medicines before and after mixing. METHODS:Mice were randomly divided into normal group,weak constitution (induced by food restriction and swimming) group,weak constitution+C. chinensis group,weak constitution+C. cassia group,weak constitu-tion+couplet medicine group,prosperous constitution (induced by high protein diet) group,prosperous constitution+C. chinensis group,prosperous constitution+C. cassia group and prosperous constitution+couplet medicine group,with 10 mice in each group. After modeling,each group was given relevant medicine intragastrically by 20 g(crude drug)/(kg·d),and normal group was giv-en equivalent volume of normal saline intragastrically for consecutive 7 days. The proportion of rats remained in high-temperature ar-eas was recorded. Organ index,biochemical index(Na+/K+-ATPase,Ca2+/Mg2+-ATPase),total antioxidant capacity(T-AOC),SOD activity,the serum content of noradrenalin and dopamine were detected. RESULTS:Compared with normal group,proportion of mice in high-temperature area increased in weak constitution group,while liver index,spleen index,renal index,biochemical in-dex activity or content decreased(P<0.05);the change of above index in prosperous constitution group were opposite to weak con-stitution group;above index had statistical significance except for the proportion of mice in high-temperature area(P<0.05). Com-pared with weak constitution/prosperous constitution group,the proportion of mice remained in high-temperature area and liver in-dex increased in weak constitution/prosperous constitution+C. chinensis group,while biochemical index activity or content de-creased(P<0.05);the change of above index in weak constitution/prosperous constitution+C. cassia group were opposite to C. chi-nensis group,spleen index and renal index increased(P<0.05);the proportion of rats remained in high-temperature area increased in weak constitution/prosperous constitution couplet medicine group,and biochemical index activity or content decreased,among which total antioxidant activity decreased significantly(P<0.05). CONCLUSIONS:The couplet medicine manifests the“cold na-ture”after C. chinensis and C. cassia equally paired,because“hot nature”degree of C. cassia is lower than“cold nature”of C. chi-nensis.
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Objective To determine the content of sophocarpine, matrine, oxysophocarpine, sophoridine, oxymatrine in Sophora Flavescentis Radix from different areas. Methods Agilent ZORBAX NH2 column (4.6 mm×150 mm, 5 μm) was used with mobile phase of acetonitrile-ethanol-3% phosphate (84∶10∶6), at a flow rate of 1 mL/min. The wavelength of detection was 210 nm. Results The linear range of sophocarpine, matrine, oxysophocarpine, sophoridine and oxymatrine were 0.022 88-0.114 4 μg (r=0.999 7), 0.083 2-0.416 0 μg (r=0.999 7), 0.376 2-1.836 0 μg (r=0.999 8), 0.104 4-0.522 μg (r=0.999 2), 0.491 2-2.456 μg (r=0.999 9), respectively. The average recovery were 101.63% (RSD=2.08%), 98.29%(RSD=1.87%), 101.89% (RSD=1.97%), 99.87% (RSD=2.06%), 102.66% (RSD=1.34%), respectively. Conclusion The method is simple, rapid and accurate, and suitable for the quality control of Sophorae Flavescentis Radix.
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AIM: To establish the method of fingerprinting analysis onZedoary turmeric oil by GC. Zedoary turmeric oil for the quality control of reference. METHODS: GC method was used to determine the fingerprint and its similarity of Zedoary turmeric oil calculated on Excel and cluster analysis. RESULTS: Seventeen samples of Zedoary turmeric oil indicated that there was a little similarity among them. Based on the cluster analysis, all the Zedoary turmeric oils could be divided into two categories, self-made and market purchasing oil, the latter oils purchased from Liaoning province and Jiangsu province varied considerably. Every category had a common fingerprint mode with clear resemblance. CONCLUSION: When finding the difference between the self-made and the marketpurchasing Zedoary turmeric oil, quality control requires serious consideration.
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Objective:To determinate the content of vitexin in Xinan Capsules by RP HPLC.Methods: C 18 ODS(4.6mm?150mm;5?m) column with temperature at 25 ?C was used with a mobile phase of water zspropanol acetic acid(75∶18∶3) elution at the flow rate of 1mL -1 ?min -1 and detection wavelength at 330nm.Results: The linear range of vitexin was from 0.734 to 3.67 ?g, and correlation coefficient was 0.9996 . The content of vitexin in Xinan Capsules is 9.7%. Conclusions:The detection method is simple and convenient, accurate with a good reproducibility. The extraction method of the control sample is simple, and easy to do.
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AIM:To establish the method for determining oridonin and rosemarinic acid in Donglingcao Tablets(Rabdosia rubescens(Hemsl.) Hara). METHODS:Oridonin and rosemarinic acid were determined by HPLC simultaneously with changing ultraviolet-visible wavelength.Chromatographic condition was composed of ODS-C_(18) column,methanol-03.% phosphoric acid (40∶60),UV detection wavelength of oridonin at 238 nm and of rosemarinic acid at 329 nm. RESULTS:The average recoveries were 97.8% and 96.7% and related standard deviations(RSD) were 1.9% and 2.3%. CONCLUSION:This method can be used to determine simultaneousely oridonin and rosemarinic acid in Donglingcao Tablets.
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AIM: The fingerprint chromatograms were established for quality evaluation of Desmodium styracifolium collected from 11 areas by HPLC. METHODS: The analysis was performed on a Phenomsil C_(18) column((4.6 mm)?250 mm,5 ?m) with acetonitrile-carbinol-water as mobile phase in a gradient mode at a flow rate of(1.0) mL/min,and at a column temperature of 25℃.The detection of wavelength was at 272 nm. RESULTS: 11 peaks were selected as the common fingerprint peaks.The relative standard deviations for relative retention values and relative peak areas were less than 3% in the precision and repeated test,the similarity of 11 batches of samples were no less than 0.9. CONCLUSION: The method is reliable and can be helpful on the quality control of Desmodium styracifolium.
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AIM: To establish methods of determing ?-eudesmol in atractylis oil from Ermiao Pills(Rhizoma Atractylodis and Cortex Phellodendri Chinensis) by gas chromatography and determing and comparing the chemical compositions between atractylis oil and poultice of Ermiao Pills by decocting separately and together. METHODS: ?-eudesmol content in Ermiao Pills decocting separtely and together was determined and compared by GC using paeonol as the internal standard substance,and contents of berberine,palmatine,jatrorrhizine in Ermiao Pills decocting separately and together were determined and compared by HPLC. RESULTS: The ?-eudesmol content in the volatile oil of decoction together was more than that of the decoction separately.But there was no remarkable difference.Berberine,palmatine,jatrorrhizine contents of decoction separately were more than that of the together. CONCLUSION: The method of determing ?-eudesmol in Ermiao Pills is established.And it has mutual effect in compatibility with Cortex Phellodendri Chinensis and Rhizoma Atractylodis.
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AIM:To simultaneously determine jatrorrhizine hydrochloride,palmatine hydrochloride,berberine hydrochloride、 evodiamine and rutaecarpine in Zuojin Pill(Rhizoma Coptidis,Fructus Evodiae).METHODS:A RP-HPLC method was established with gradient elution and an agilent C_ 18 column(250 mm?4.6 mm,5 ?m)was used.The mobile phase A was acetonitrile and the mobile phase B was water with 0.3% phosphoric acid-triethylamine.The detective wavelength was at 225 nm.RESULTS:Jatrorrhizine hydrochloride had a good linearity in the range of 0.013-0.065 ?g.The average recovery was 103.1% with RSD of 1.1%.Palmatine hydrochloride had a good linearity in the range of 0.013-0.102 ?g.The average recovery was 99.8% with RSD of 1.9%.Berberine hydrochloride had a good linearity in the range of 0.063-0.317 ?g.The average recovery was 99.8% with RSD of 1.4%.Evodiamine had a good linearity in the range of 0.001-0.007 ?g.The averoge recovery was 100.5% with RSD of 1.5%.Rutaecarpine had a good linearity in the range of 0.002-0.010 ?g.The average recovery was 98.9% with RSD of 1.9%.CONCLUSION:This method is simple,accurate and reproducible.It can be used to determine jatrorrhizine hydrochloride、 palmatine hydrochloride、 berberine hydrochloride、 evodiamine and rutaecarpine together with the same chromatogram condition.
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AIM: To use seven parts cortex phellodendri and one part Rhizoma atractrylodis to extract mixedly,and study mouse's pharmacokinetic parameter of berberine derived from extract. METHODS: Evodiamine was taken as the marker,HPLC was used to determine berberine content in rat plasma. RESULTS: The linear range of rat's plasma sample was within 0.006 6-0.211 2 ?g/mL,RSD was little than 10%,detection limit was 0.0033 ?g/mL.The main pharmacokinetic parameters consisted of 2 h(T_max),0.183 mg/L(C_max),10.87 h(MRT_0-24),21.562?4.448(MRT_0-∞). CONCLUSION: Berberine in combination of cortex phellodendri and Rhizoma atractylodis can be quickly absorb and achieve the peak value,but metabolize slowly.
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AIM: To establish the method of fingerprinting analysis on Zedoary turmeric oil by GC.Zedoary turmeric oil for the quality control of reference. METHODS: GC method was used to determine the fingerprint and its similarity of Zedoary turmeric oil calculated on Excel and cluster analysis. RESULTS: Seventeen samples of Zedoary turmeric oil indicated that there was a little similarity among them.Based on the cluster analysis,all the Zedoary turmeric oils could be divided into two categories,self-made and market purchasing oil,the latter oils purchased from Liaoning province and Jiangsu province varied considerably.Every category had a common fingerprint mode with clear resemblance. CONCLUSION: When finding the difference between the self-made and the market purchasing Zedoary turmeric oil,quality control requires serious consideration.