Effects of Quercetin Extracted from Flower Buds of Sophora japonica cv.jinhuai on Proliferation and Apoptosis of Human Breast Cancer MCF-7 Cells

[Objectives]To investigate the effects of quercetin extracted from flower buds of Sophora japonica cv.jinhuai on the proliferation,apoptosis and migration of human breast cancer MCF-7 cells.[Methods]MTT assay,inverted microscope observation,hoechst33342 staining,flow cytometry(FCM)and wound healing assay were adopted to investigate the proliferation,morphological changes,apoptosis level and cell migration ability of human breast cancer MCF-7 cells,respectively.[Results]The morphological changes of cells in the treatment groups included gradually decreased number,reduced volume,vague cell contour,loose intercellular connection,uneven cytoplasm distribution and increased cell debris.With the increase of drug concentration,quercetin significantly inhibited the proliferation of human breast cancer MCF-7 cells(P<0.05).The number of apoptotic bodies increased gradually.When the concentration reached 100μmol/L,a large number of nuclear fragments appeared,and the level of apoptosis was statistically different(P<0.05).The mobility and migration ability of cells showed a decreasing trend,and the differences were statistically significant(P<0.05).[Conclusions]This study can provide experimental basis for clinical application of quercetin against breast cancer.

Antioxidant and Hypoglycemic Ability of Ardisia gigantifolia Stapf Parts

[Objectives]To study the antioxidant and hypoglycemic effects of different parts of Ardisia gigantifolia Stapf.[Methods]The hydroxyl radical scavenging activity,DPPH radical scavenging activity and total antioxidant capacity of ABTS of 75%ethanol extract of A.gigantifolia Stapf and the petroleum ether,ethyl acetate,n-butanol,chloroform and aqueous extract were measured with Vc as positive control.At the same time,acarbose was used as reference substance to determine the inhibitory effect of each polar site onα-glucosidase.[Results]All parts of A.gigantifolia Stapf had antioxidant activity,among which ethyl acetate had the strongest antioxidant activity,and the scavenging rate of hydroxyl radical and DPPH radical was higher than that of positive control.The results showed that petroleum ether,ethyl acetate and chloroform had a good inhibitory effect onα-glucosidase(better than acarbose).[Conclusions]The ethyl acetate part of A.gigantifolia Stapf had the best antioxidant activity and inhibitory effect onα-glucosidase.It provides a basis for further research and development of A.gigantifolia Stapf.

Method for Improving the Determination of Dibromoacetic Acid in Cefathiamidine

[Objectives]To establish an LC-MS method for the determination of dibromoacetic acid in Cefathiamidine and Cefathiamidine for Injection.[Methods]Shiseido PC HILIC column(2.0 mm ×100 mm,3μ.m)was used.The mobile phase was 0.005 Mammonium formate and the mobile phase was acetonitrile with gradient elution.The flow rate was 0.3-0.8mL/min.[Results]The limit of detection was 0.2500 ng/mL and the limit of quantification was 0.500 ng/mL,which were 2.5%and 5.0%of the impurity limits,respectively.The recov-ery rate was 103.85%.[Conclusions]This method improved the detection of dibromoacetic acid impurities in Cefathiamidine,and has that advantage of good specificity,low limit of detection and limit of quantification,high sensitivity,high accuracy,interference resistance,can meet the detection requirements of Cefathiamidine,and is suitable for the quality control of Cefathiamidine.

Optimization of Solid-state Fermentation Conditions of Sophora japonica cv.jinhuai by Response Surface Methodology

[Objectives]To optimize the solid-state fermentation process of Flos Sophorae Immaturus by Penicillium with Sophora japonica cv.jinhuai as raw material.[Methods]The fermentation conditions were optimized by single factor experiment and response surface methodology with quercetin content as the dependent variable.[Results]According to the established model,the optimal fermentation process of Flos Sophorae Immaturus was obtained as follows:temperature 29.97℃,time 6.88 d,rotation speed 180.86 rpm,inoculation amount 3.93 mL,and the expected content of quercetin was 34.8053 mg/g.Based on this,the fermentation parameters were adjusted,and the actual content was 33.67 mg/g,which was close to the predicted value.[Conclusions]The optimization of fermentation process of Flos Sophorae Immaturus by response surface methodology provides a reference for the development and utilization of this medicinal material.

Effects of Polysaccharide from Polygala fallax on Apoptosis of B16 Melanoma

[Objectives]The paper was to study the effects of polysaccharide from Polygala fallax on apoptosis of B16 melanoma and its mechanism.[Methods]P.fallax polysaccharide was prepared by water extraction and alcohol precipitation method combined with Sevag method.The effects of different concentrations of P.fallax polysaccharide on B16 cell proliferation were detected by MTT assay.Apoptosis was detected by Hoechst 33258 staining assay and flow cytometry.The expressions of Bcl-2,Bax,Cleaved Caspase-3 mRNA were detected by Real-time PCR assay.The expressions of Bcl-2,Bax,Caspase-3,Caspase-8,Caspase-9,Cleaved Caspase-3,Cleaved Caspase-9,ERK,p-ERK,JNK,p-JNK proteins were detected by Western Blot assay.[Results]P.fallax polysaccharide inhibited the cell proliferation of B16 melanoma in a concentration-dependent manner(24 h IC 50=0.2133 mg/L;48 h IC 50=0.08489μg/mL).P.fallax polysaccharide up-regulated the expressions of Bax,Cleaved Caspase-3 mRNA and protein,down-regulated the expressions of Bcl-2 mRNA and protein,activated Caspase-3,Caspase-8 and Caspase-9,and decreased the levels of p-ERK/ERK and p-JNK/JNK.[Conclusions]P.fallax polysaccharide can inhibit the cell proliferation of B16 melanoma and induce their apoptosis,probably attributed to the regulation of MAPK,Bcl-2 and Caspase family signaling pathways.