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[Objective]To investigate the mechanism of Compound Fuling Granule(CFG)in inhibiting the glucose metabolism and metastasis of ovarian cancer cells.[Methods]Ovarian cancer cell HEY-T30 were cultured in vitro and incubated with different concentration(0.25,0.5,1,1.5,2 mg·mL-1)of CFG for 24 h.Cell counting kit-8(CCK-8)assay was performed to detect the effect of CFG on the cell proliferation,Transwell assay was used to investigate cell migration ability,lactic acid detection kit and glucose detection kit were used to detect lactic acid production and glucose consumption,the expressions of dynamin-related protein 1(DRP1),some key glycolysis-related proteins and metastasis-related proteins were detected by Western blot,Mito-Tracker Red was used to label mitochondria to observe mitochondria morphology.Lentivirus transfection technique was used to achieve the stable DRP1 knockdown HEY-T30 cells.Real-time quantitative polymerase chain reaction(Real-time qPCR)was used to detect the expression of DRP1 mRNA,the effect of CFG on lactic acid production and glucose consumption of HEY-T30 after DRP1 knockdown was detected by lactic acid detection kit and glucose detection kit,Transwell assay was used to investigate the migration ability of HEY-T30 with DRP1 knockdown after treated with CFG,the effect of CFG on the expression of glycolysis-related proteins and metastasis-related proteins in HEY-T30 with DRP1 knockdown was detected by Western blot.[Results]Compared with control group,the cell survival rate in 0.5,1,1.5,2 mg·mL-1 CFG groups reduced significantly(P<0.01).The average length of mitochondria in 0.5,1 mg·mL-1 group was markedly increased(P<0.01),the protein expression of DRP1 was significantly reduced(P<0.05,P<0.01),and the lactate production and glucose consumption in 0.5 and 1 mg·mL-1 groups were significantly decrease(P<0.01).The number of migration cell was significantly reduced in 0.25,0.5 and 1 mg·mL-1 concentration groups(P<0.05,P<0.01).After knockdown of DRP1,the glycolysis level and migration of HEY-T30 were decreased(P<0.05,P<0.01),and the expressions of glycolysis-related proteins and metastasis-related proteins were decreased(P<0.05,P<0.01).The inhibitory effect of CFG on glycolysis and metastasis of ovarian cancer cells was also weakened.[Conclusion]By targeting DRP1 to regulate glucose metabolism reprogramming,CFG could inhibit the metastasis of ovarian cancer.
RESUMO
Objective:To explore the clinical characteristics and distribution of Tranditional Chinese Medicine (TCM) syndrome types of refractory tic disorders in children based on clustering analysis.Methods:A cross-sectional study. 183 children aged 3-18 years with refractory tic disorders from pediatric encephalopathy outpatient department of the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine who met the inclusion criteria from October 2015 to January 2022 were recruited. Frequency descriptive analysis, systematic clustering analysis and principal component analysis were conducted by Microsoft Office Excel 2016 and IBM SPSS Statistics 26.0 to explore the clinical characteristics and syndrome distribution of the disease.Results:The TCM symptoms of 183 children with refractory tic disorders were clustered into 6 categories, and finally 5 types of syndrome were obtained, among which the syndrome of spleen deficiency and liver hyperactivity mixed with dampness accounted for the largest proportion (27.32%), followed by the syndrome of phlegm-fire disturbing spirit (21.31%), the syndrome of wind-heat invading lung (18.03%), the syndrome of phlegm-qi stagnation (17.49%), and the syndrome of dampness and yin deficiency (15.85%).Conclusion:Through the mining and analysis of the outpatient information of pediatric encephalopathy in our hospital, the common syndrome differentiation types of refractory tic disorder are obtained, which can provide a reference for the TCM syndrome differentiation types of refractory tic disorder in children.