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【Objective】 To establish an enzyme-linked immunosorbent assay (ELISA) method for the determination of residual human coagulation factor Ⅺ in human prothrombin complex and validate the method. 【Methods】 Human factor Ⅺ was reacted with the capture antibody coated on the microtiter plate. After appropriate washing steps, biotinylated primary antibody was bound to the captured protein. Excess primary antibody was washed away and bound antibody was reacted with horseradish peroxidase conjugated streptavidin. TMB substrate was used for color development at 450 nm. The dilution reliability, accuracy, specificity, repeatability, intermediate precision, linearity, range and durability were verified. 【Results】 The verification results showed that the accuracy and specificity of this method met the experimental requirements, with an average recovery rate of 109.2% and RSD of 6.93%. The repeatability RSD was 6.78%, and the intermediate precision RSD was 6.75%, indicating good precision. The linear regression correlation coefficient of standard curve was 0.999 9, showing good accuracy and precision within the linear range. The durability was verified by the incubation time and the validity period of reagent kit opening. The results showed that the RSD of the incubation time change was 6.62%, indicating that the incubation time of this detection method was controlled between 28 to 32 minutes, and there was no significant impact on the results. The RSD of the detection results before and after the reagent kit was opened and stored under conditions for 7 days was 3.84%, indicating that the preservation of the reagent kit according to the conditions for 7 days after opening has no effect on the FⅪ detection results. Both indicated that the method had good durability. The dilution reliability results showed that there was a "hook" effect in the detection of FⅪ residue in human prothrombin complex, which could be solved by diluting 100 to 200 times. 【Conclusion】 This method can be used for the determination of FⅪ residues of human prothrombin complex in laboratory.
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Objective To investigate the relationship between CYP 2C19 gene polymorphisms and clopidogrel efficacy in coronary heart disease patients after percutaneous coronary intervention (PCI).Methods From January 2016 to December 2017,62 patients with acute coronary syndromes and treated with PCI in Guizhou Aerospace Hospital were recruited, CYP2C19 genotype, ADP －induced platelet aggregation rate and myocardial enzymes and other indicators were detected before operation .The myocardial enzymes were measured 24 hours after PCI.According to different metabolic types,the patients were grouped,the above indicators were compared.Results The CYP2C19*1/*1 was 37.10%,CYP2C19*1/*2 was 35.48%,CYP2C19*1/*3 was 11.29%,CYP2C19*2/*2 was 12.90%,CYP2C19*2/*3 was 3.23% and CYP2C19*3/*3 was 0.00%.The LDH,AST,CK,CK－MB and α－HBDH in the PCI patients after operation were significantly higher than those before operation (t＝0.019,0.040, 0.044,0.022,0.014,all P＜0.05).But ADP induced platelet aggregation rate and myocardial enzymes and other indicators among fast metabolism group,intermediate metabolic group and slow metabolic group had no statistically significant differences (all P ＞0.05).Conclusion CYP2C19 mutation frequency in the Chinese population is relatively large,the sample size of this study is less ,the relationship between clopidogrel resistance and the specific genotype can not be obtained ,it need to increase the sample size and comprehensive multi －factor consideration .
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Objective:To study the effect of miR-122a on inhibition proliferation of laryngeal carcinoma cell line Hep-2.Meth-ods:The oligomucleotide of miR-122a was transfected into laryngeal carcinoma cell line Hep 2 cells,which were devided into three groups of A(miR-122a transfection),B(miR-122a inhibitor),C(miR-122a-NC inhibitor) and group of D(blank control).The expres-sion of miR-122a was defected by RT-PCR,and relevant protein expression was evaluated by Western blot .The cell proliferation and cell cycle were determined by MTT assy and flow cytometry ,respectively.Results:Compared to group D,miR-122a expression in Hep2 cells was obviously elevated atter miR-122a-transfected.The proliferation of Hep2 cells in group A was significantly inhibited and the cell cycle arrested at G1/G0 phase.The protein expression of CDC42 was downregulated with decreased expressions of CDK 4 and cyclin D1 in group A.Conclusion:miR-122a inhibits the proliferation activity of Hep 2 cells,suggesting that miR-122a can be taken as a po-tential candidate for gene therapy of laryngeal carcinoma .
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Composition of invention patent application is important for obtaining patent right and maintaining the right stability. According to the features of TCM invention patent application and common problems in examination, this article summarized several key points for effective application composition, including composing background technology objectively, highlighting technical problems, reasonable layout of technical pian, working in contact with technical effects, and clarifying technical terms. It expounded each point with relevant typical cases, with a purpose to provide references for applicants and agents of TCM invention patent application.
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<p><b>OBJECTIVE</b>To establish a HPLC method to determine the content of aromadendrin in Euonymus alatus.</p><p><b>METHOD</b>Kromasil 100-5C18 column was used with a mobile phase composed of acetonitrile-0.3% glacial acetic acid (24:76) at a flow rate of 1.0 mL x min(-1). The detection wavelength was 292 nm and the temperature of column was 30 degrees C.</p><p><b>RESULT</b>The calibration curve was linear in the range of 0.051-2.04 microg for aromadendrin. The correlation coefficient of the calibration curve was 0.999 9. Twenty batches of the crude E. alatus purchased from different areas were determined and the contents of aromadendrin in the twigs of E. alatus were fluctuated from 0.001 57% to 0.041 6%.</p><p><b>CONCLUSION</b>This method is simple, repeatable and can be used for determination of aromadendrin.</p>
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Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Chemistry , Euonymus , Chemistry , Flavonoids , Plant Extracts , Chemistry , Plant Stems , ChemistryABSTRACT
Objective To investigate the situation of the activities of daily living(ADL)for elderly people in Changsha,and analyze the influencing factors. Methods Questionnaire survey on ADL was held in 2239 elderly people(ranging from 60 to 91 years old),and the influencing factors including age,sex,family structure,marital status,education level,occupation,common chronic diseases and medical care consciousness were analyzed. Results Going to the toilet irregularly hadthe highest injury rate in physical self-maintenance viability(18.2%).But in instrumental ADL,the highest injury rate occurred while shopping(26.3%).Advanced age,living alone,losing spouse,sickness and the lack of medical care awareness were the major factors that influenced the ADL of the elderly people. Conclusions The ADL is an important indicator which reflects the health condition of elderly people.Advanced age,living alone,losing spouse,having many chronic diseases and lacking in medical care awareness are risk factors.
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Objective To develop a HPLC quantitatively test method of glycyrrhizin, liquiritin and isoliquritigenin simultaneously for licorice. Methods The 67% methanol extracts of licorice powder were analyzed by reverse-phase HPLC on an ODS column, using a gradient elution with a mixture of 1% H3PO4 and acetonitrile. The eluate was monitored by UV absorption at 248, 276, 360 and 370 nm. Results The linear equation of glycyrrhizin was Y =1?106X +16 220, r 2=1. The linear equation of liquiritin was Y= 2?106X+49 444, r 2=0.999 5. The linear equation of isoliquritigenin was Y =1?107X +4.466 7, r 2=1. The recovery rate of glycyrrhizin, liquiritin and isoliquritigenin was 98.01%, 102.63% and 98.18% respectively. Conclusion The method is accurate, stable amd reliable, and can be used to determine the three main chemical contents simultaneously.
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Objective To study the protective effect of Duzhong Hongjingtian Capsule (DHC) on cerebral ischemia in rats. Methods Thrombotic ischemia and middle cerebral artery occlusion (MCAO) induced by FeCl3 in rats was used to observe the protective effect of DHC on ischemia stroke, and common carotid artery occlusion (CCAO) rats was used to analyze the effect of DHC on blood brain barrier (BBB) permeability. Results DHC could improve the abnormal behavior of MCAO rats and decrease the infarction area and water content in the brain of MCAO rats. DHC could also improve the cerebral blood flow on MCAO rats and significantly reduce the permeability for evans' blue (EB) of CCAO rats. Conclusions DHC can protect the brain ischemic injury.