摘要
Objective To detecte the differential metabolites and related pathways in Siha cells of cervical cancer by screening the inhibition of HPV16 E6/E7 expression based on 1H NMR metabolomics so as to identify the key metabolic markers involved in the development of high-risk HPV16 cervical cancer.Methods Siha cells were transfected with RNAi fragments to down-regulate the expression of E6/E7,which were divided into the normal control group(Siha cells),no-load group(si-NON),si-E6 group and si-E7 group,and their transfection efficiency was verified.1H NMR metabolomics was used to reveal the differential metabolites involved in interfering E6/E7 expression in Siha cells.Combined with MetaboAnalyst 5.0 online software,differential metabolites and related metabolic pathways were obtained.Results Fluorescence was observed by inverted fluorescence microscope.Western blotting results showed that compared with Siha group,the expression of E6/E7 in si-E6 group and si-E7 group was decreased(F=145.8,P<0.001).After down-regulating the expression of E6/E7,13 common differential metabolites,including Isoleucine,Leucine and valine,were detected.The results of MetaboAnalyst 5.0 online software analysis suggested that the above metabolites were mainly involved in the biochemical synthesis pathway of aminoacyl-trNA,biochemical synthesis pathway of isoleucine,Leucine and valine;There were 10 metabolic pathways of tyrosine,phenylalanine and tryptophan biochemical synthesis.Conclusion After HPV16 infection,changes of glucose and amino acid metabolism can promote the progression of cervical cancer,which provide a theoretical basis for the prevention and treatment of cervical cancer.
摘要
Objective To establish an HPLC quantitative method for the content determination of baicalin in Kangdeling capsule .Methods The chromatography column was Agilent Tc-C18-WR (4 .6 mm × 250 mm ,5μm) ,and the column temper-ature was 30 ℃ .The mobile phase consisted of acetonitrile and 0 .5‰ phosphoric acid (26 ∶ 74 ) .The flow rate was 1 .0 ml/min ,and the detection wavelength was 265 nm .Results The retention time of baicalin was about 16 min .The calibra-tion equation was Y=22 114 .67 X -112 836 .7(r=0998 8)with good linearity in the range of 5 .410-108 .2 μg/ml for baicalin . The average recovery was 98 .78% while RSD were 0 .74% .Conclusion This method is simple ,time-saving and accurate which could be used to routine analysis of baicalin in Kangdeling capsule .
摘要
Objective To develop a method for determination of the rosmarinic acid in Xiaokang capsules .Methods An HPLC method was developed .The determination was performed on Agilent C18 column at 30℃ with a mobile phase composed of methanol :0 .5% formic acid (50∶50) run at a flow rate of 1 .0 ml/min .The detection wavelength was set at 330 nm and the injection volume was 10 μl .Results The linear range of rosmarinic acid concentrations was 2 .88 to 20 .1μg/ml (r=0 .999 8) . The average recovery was 99 .5% (RSD=0 .40% ,r=9) .Conclusion An assay for the determination of the rosmarinic acid in Xiaokang capsule that is simple ,rapid ,accurate and reliable has been developed .