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OBJECTIVE To study the improvement effect of total flavonoids from Rosa multiflora root on vascular injury in rheumatoid arthritis （RA） model rats and its potential mechanism. METHODS Female Wistar rats were randomly divided into normal control group， model group， aspirin group （positive control， 30 mg/kg）， low-dose and high-dose groups of total flavonoids from R. multiflora root （4.15， 8.30 g/kg， by crude drug）， with 10 rats in each group. Except for the normal control group， the RA model was induced in other groups by collagen induction and high-fat diet. After 14 days of modeling， they were given corresponding drug solution/0.5% sodium carboxymethyl cellulose solution intragastrically， once a day， for 36 consecutive days. The total body score， arthritis index （AI） and swollen joint count （SJC） of the rats were evaluated regularly. After the last medication， serum levels of interleukin-6 （IL-6）， intercellular adhesion molecule-1 （ICAM-1） and vascular cell adhesion molecule- 1 （VCAM-1） were determined. The pathological morphological changes in the vascular tissue of thoracic aorta were observed； the protein expression of Toll-like receptor 4 （TLR4） and the protein phosphorylation levels of Janus kinase 2 （JAK2） and signal transduction and activator of transcription 3 （STAT3） in vascular tissue of thoracic aorta were measured. RESULTS Compared with the normal control group， serum levels of IL-6， ICAM-1 and VCAM-1， protein expression of TLR4， and the protein phosphorylation levels of JAK2 and STAT3 in vascular tissue of thoracic aorta were increased significantly in model group （P＜ 0.01）. The atherosclerotic plaque （atheroma）， cholesterol crystal， lymphocyte infiltration and a small number of unbroken foam cell aggregation could be seen in the vascular tissue of thoracic aorta. Compared with the model group， total body score （except for the low-dose group）， AI and SJC were decreased significantly in groups of total flavonoids from R. multiflora root on the 28th day （P＜0.05 or P＜0.01）； total body score，AI and SJC were decreased significantly in low-dose group of total flavonoids from R. multiflora root on the 49th day （P＜0.05 or P＜0.01）； the other quantitative indicators in serum and vascular tissue were significantly reversed in groups of total flavonoids from R. multiflora root （P＜0.05 or P＜0.01）， and pathological damage of vascular tissue was significantly relieved. CONCLUSIONS Total flavonoids from R. multiflora root can significantly improve vascular injury in RA model rats， and its mechanism may be related to reducing the protein expression of TLR4 in vascular tissue and inhibiting the activation of IL-6/JAK2/ STAT3 signaling pathway.
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OBJECTIVE： To establish content determination method for simultaneously determining aucubin，geniposidic acid，catechin，chlorogenic acid，asperuloside，rutin，isoquercitrin and astragalin in Eucommia ulmoides leaves. METHODS：HPLC method was adopted. The determination was performed on Agilent ZORBAX SB-C18 column with the mobile phase consisted of acetonitrile-0.1％ phosphoric acid（gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 203 nm for aucubin and catechin，239 nm for geniposidic acid and asperuloside，220 nm for chlorogenic acid，354 nm for rutin and isoquercitrin，and 266 nm for astragalin. The sample size was 5 μL. RESULTS： The linear range of aucubin， geniposidic acid， catechin， chlorogenic acid， asperuloside， rutin， isoquercitrin and astragalin were 0.812-6.090 μg（r＝0.999 3），0.438-3.285 μg（r＝0.999 2），0.045-0.336 μg（r＝0.999 2），0.882-6.615 μg（r＝0.999 3），0.097-0.726 μg（r＝0.999 1），0.064-0.483 μg（r＝0.999 3），0.048-0.360 μg（r＝0.999 1） and 0.014-0.108 μg（r＝0.999 7），respectively. RSDs of precision， stability and reproducibility tests were all lower than 3.5％（n＝6）. The average recovery rates were 101.60％，103.06％，99.77％，96.93％，98.17％，96.75％，98.97％ and 99.60％，with RSDs of 1.42％，2.65％，2.78％，2.05％，2.26％，0.93％，2.79％ and 3.08％，respectively（n＝6）. The contents of aucubin， geniposide， catechin， chlorogenic acid， plantain， rutin， isoquercetin and astragaloside in 12 batches of E. ulmoides leaves from different collection time and planting varieties were 10.903-17.245， 5.578-7.892， 0.198-0.440， 13.890-19.782， 1.008-1.547， 1.102-2.396， 0.267-0.701， 0.150-0.412 mg/g， respectively. The content fluctuated greatly. The contents of aucubin， geniposide， catechin， chlorogenic acid， rutin and pinoresinol diglucoside in cortex of E. ulmoides were 0.299， 0.123， 0.580， 0.112， 0.026，1.961 mg/g， respectively. CONCLUSIONS： The method is simple， reproducible and accurate. It can be used to evaluate the quality of E. ulmoides leaves. There are obvious differences in composition and content of components in different medicinal parts （cortex， leaves） of E. ulmoides.
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OBJECTIVE： To study improvement effects of different proportions of total glucosides of ginseng （TGG）， total glucosides of moutan cortex （TGM） and paeonol containing serum on the injury of human umbilical vein endothelial cells （HUVEC） injury induced by hydrogen peroxide （H2O2）， screen the optimal proportion and investigate its mechanism. METHODS： The rats were randomly divided into blank group （distilled water）， TGG group （TGG， 2.025 g/kg）， TGM group （TGM， 4.05 g/kg） and paeonol group （paeonol， 1.08 g/kg）， with 12 rats in each group. They were given relevant medicine twice a day for consecutive 7 days. 1 h after last medication， the blood samples were collected via abdominal aorta to prepare drug containing serum. Using survival rate of HUVEC as evaluation indexes， different proportions of TGG， TGM and paeonol containing serum as factors， L9（34） orthogonal test was designed to optimize the optimal proportion of 3 kinds of drug containing serum. HUVEC were divided into blank group， model group， TGG group， TGM group， paeonol group and optimal proportion group. Except that blank group were treated with relevant medium， other group were treated with 1.2 mmol/L H2O2 to induce HUVEC injury， and then TGG group （volume fraction of drug containing serum was 0.000 5％）， TGM group （volume fraction of drug containing serum was 0.000 5％）， paeonol group （volume fraction of drug containing serum was 1％） and optimal proportion group were intervened with drug containing serum. The levels of LDH， NO and ET-1 in cells were detected by microplate method and ELISA. RESULTS： The optimal proportion of drug containing serum were TGG 0.000 5％， TGM 0.000 5％ and paeonol 1％. Compared with blank group， the levels of LDH and ET-1 were higher in model group （P＜0.01）， while NO level was lower （P＜0.05）. Compared with model group， the levels of NO were higher in TGG group， TGM group and optimal proportion group （P＜0.01）， while the levels of LDH and ET-1 were lower （P＜0.05 or P＜0.01）. Compared with TGG group， TGM group and paeonol group， the level of LDH was lower in optimal proportion group （P＜0.05 or P＜0.01）， while the level of NO was higher （P＜0.05 or P＜0.01）. CONCLUSIONS： TGG and TGM combined with paeonol can significantly improve HUVEC injury induced by H2O2， and the mechanism of which may be associated with the decrease of LDH and ET-1 and the increase of NO.
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OBJECTIVE:To optimize the processing technology of rehmanniae radix praeparata. METHODS:Using transfer rates of catalpol,rehmaionoside D,acteoside,isoacteoside,polysaccharide as indexes for comprehensive score,heating tempera-ture(pressure),heating time and heating times as investigating factors,L9(34)orthogonal test was used to optimize the processing technology of rehmanniae radix praeparata,and verification test was conducted. RESULTS:The optimal processing technology of rehmanniae radix praeparata was as follow as heating temperature of 125 ℃,pressure of 150 kPa for twice,2 h every time. The comprehensive scores of 3 batches of samples were 0.6985,0.6755,0.7016 in the verification test,respectively,RSDs were less than 5%(n=3). CONCLUSIONS:Optimized processing technology is simple,stable,feasible,and can provide reference for in-dustrial production of rehmanniae radix praeparata.
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Objective To develop a HPLC method for the determination of ephedrine hydrochloride in Xiaochuanning capsules.Methods A Hypersil ODS column was used.The mobile phase was acetonitrile-water solution of 0.05 mol / L NaH2PO4(pH2.7,22 ∶ 78).The flow rate was 1.0 mL / min and column temperature was 40 ℃.Detcted wavelength was 205 nm.Results A good linearity was in the range of 0.1005 ~ 0.5025 ?g(r = 0.9993).The average recovery was 103.1 %,RSD=1.81 %.Conclusion The method is simple,accurate,and can be used for the quality control of Xiaochuanning capsules.
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TLC scanning technique was used for the determination of tanshinone ⅡA in "Prostato Healthcare Bag" at ?S=470nm, and ?R = 620nm. This method is simple and highly sensitive, with an average recovery of 97.45％ and a variation coeffioient of 1,28%.
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Objective To establish the quality standard for Xiaoyao Effervescent Tablets.Methods Radix Paeoniae Alba,Radix Angelicae Sinensis,Radix Bupleuri and Radix Glycyrrhizae in the tables were identified by TLC.Paeoniflorin content was determined by HPLC.Results Radix Paeoniae Alba,Radix Angelicae Sinensis,Radix Bupleuri and Radix Glycyrrhizae could be identified by TLC,and the identification was highly specific without interference.The linear range of paeoniflorin was 0.082 ? g～ 1.025 ? g.The average recovery was 100.44 %(RSD=2.37 %).Conclusion The method is simple,reliable,and accurate,and can be used for the quality control of Xiaoyao Effervescent TabLets.
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Objective To develop a HPLC method for the determination of scutellarin in Brevisapin Tablets.Methods C18 column with temperature of 40 ℃ was used.The mobile phase was the mixed solution of methanol-water-phosphoric acid(40 ∶ 60 ∶ 0.5) with the flow rate of 1.0 mL? min-1.Determination wavelength was 335 nm and the sample injected volume was 10 ? L.Results A good linearity of scutellarin was in the rang of 0.161 5～ 0.807 5 ? g.The average recovery of scutellarin was 98.7 % with RSD=1.26 %.Conclusion The method is simple,accurate,and can be used for the quality control of Brevisapin Tablets.
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Objective To develop a HPLC method for the determination of protopine in Rhizoma Corydalis Decumbentis Capsules. Methods A HPLC was performed on a Hypersil ODS column( 4.6? 250 mm, 5 ? m) . The mobile phase was methanol - 0.1 % water solution of triethylamine (65∶ 35). Determination wavelength was 290 nm and flow rate was 1.0 mL/min. The theoretical plates should over 4000 according to the content of protopine. Results A good linearity was shown in the range of 0.3168～ 1.5840 ? g, r=0.9996. The average recovery was 100.6 % , RSD=2.68 % . Conclusion The method is simple, effective and can be used for the quality control of Rhizoma Corydalis Decumbentis Capsules.
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AIM: To establish the HPLC method for determinating syringin and betaine in Qianglining Tablet(Radix et Rhizoma seu Caulis Acanthopanacis senticosi,Fructus Lycii). METHODS: A Hypersil C_18 column was used for the determination of syringin.The mobile phase for syringin was methanol-0.1 mol/L acetic acid (24∶76),and the detection wavelength was at 270 nm.A Hypersil NH_2 column was used for the determination of betaine.The mobile phase for betaine was acetonitrile-water(25∶75),and ELS detector was used. RESULTS: The linear range of syringin and betaine were 0.121 6-0.608 0 ?g(r=0.999 9) and 0.660 8-3.964 8 ?g(r=0.999 1),respectively.The average recoveries of syringin and betaine were 99.09%(RSD=2.13%) and 96.34%(RSD=1.31%),respectively. CONCLUSION: The method is simple,accurate,and can be used for the quality control of Qianglining Tablet.
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AIM: To establish a HPLC method for determining of ginsenoside Re,ginsenoside Rg_1 and schisandrin in Huoliyuan Tablets(total ginsenoside,Radix Astragali,Fructus Schisandrae Chinensis,Radix Ophiopogonis,etc.). METHODS: A Hypersil C_(18) column was used.The mobile phase for ginsenoside Re and Rg_1 was acetonitrile-water mixed with 0.05 mol/L NaH_2PO_4(pH2.7,20∶80),and determination wavelength was 203 nm.The mobile phase for schisandrin was acetonitrile-0.1%H_3PO_4(48∶52),and determination wavelength was 250 nm. RESULTS: The linear ranges of ginsenoside Re,Rg_1 and schisandrin were 1.84-14.72 ?g(r=0.999 4),1.49-11.92 ?g(r=0.999 0) and 0.094-0.47 ?g(r=0.999 9),respectively.The average recoveries of ginsenoside Re,Rg_1 and schisandrin were 99.86%(RSD=1.69%),98.17%(RSD=2.03%) and 101.54%(RSD=(2.62%),) respectively. CONCLUSION: The method is simple,accurate,and can be used for the quality control of Huoliyuan Tablets.