1.Let-7a/g/i targeted to Aurora-B in human osteosarcoma cells
Yunfei ZHOU ; Jiaming LIU ; Xuanyin CHEN ; Liangbo ZHU ; Xinhua LONG ; Yang ZHOU ; Zhihong ZHANG ; Zhili LIU
China Oncology 2015;(12):966-971
Background and purpose:MicroRNA(miRNA) is a class of small non-coding RNA playing an important regulatory role in many tumors. This study investigated which miRNA might negatively regulate the expression of Aurora-B in osteosarcoma cells, and to lay the foundation for the further investigation of the effort and regulation of Aurora-B in osteosarcoma malignant phenotype.Methods:Bioinformatics prediction software (http://www.targetscan.org) and luciferase assays were used to investigate which miRNA might target to modulate the Aurora-B. Real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot assay were used to further verify which miRNA could negative regulate the expression ofAurora-B gene.Results:Bioinformatics prediction showed let-7 family have the possibility to modulate the expression of Aurora-B; Luciferase assays showed thatAurora-B might be the target gene of let-7a/b/c/d/e/f/g/i; RTFQ-PCR and Western blot analysis testiifed that both the expression levels of Aurora-B mRNA and Aurora-B protein were signiifcantly decreased in Let-7a/g/i up-regulated U2-OS and HOS cells, compared to the cells in the negative control group; but in Let-7b/c/d/e/f up-regulated U2-OS and HOS cells, the expression levels of Aurora-B mRNA and Aurora-B protein have no signiifcant difference, compared to the cells in the negative control group.Conclusion:Let-7a/g/i may downregulate the expression of Aurora-B in human osteosarcoma cells.
2.Network Pharmacological Analysis and Experimental Verification of Huayu Xiaozheng Granule in the Treatment of Ectopic Pregnancy Based on JAK/STAT Pathway
Shuo YUAN ; Xuanyin WANG ; Jiaqi ZHANG ; Litian XU ; Zhenyu HU ; Gaopi DENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(9):3011-3023
Objective To investigate the underlying mechanism of Huayu Xiaozheng granule(HYXZ)in the treatment of ectopic pregnancy(EP).Methods The potential ingredients and targets of HYXZ were identified and screened.PPI network,GO enrichment and KEGG pathway analysis were established to understand the potential and intricate mechanism of HYXZ in treating EP.The effects of HYXZ on HTR-8/SVneo cells migration and invasion were detected by wounding healing assay and transwell migration assay respectively,as well as the influence of HYXZ granule on JAK/STAT pathway and associated proteins were assessed by western blot under the control of JAK/STAT pathway inhibitor and agonist.Results A total of 375 active ingredients,45 known targets and 207 predicted targets of HYXZ were obtained.A PPI network has constructed to know the relationship of 252 common targets of drug targets and disease targets,from which we screened out 15 key targets:STAT3,MMP9,TLR4,HIF1A and others.Scratch test showed that HYXZ inhibited the migration of the HTR-8/SVneo cells cells in a time and dose-dependent manner;Transwell test showed that the number of transmembrane cells decreased significantly(P<0.05)after 48 h with various concentrations of HYXZ;Western blot experiment showed that the expression of JAK/STAT pathway protein decreased significantly(P<0.05);and the expression of its related proteins uPA,HIF1and N-cadherin were inhibited(P<0.05),the tendency of which was consistent with the inhibitor AG490,but opposite to the agonist IL-6.Conclusion HYXZ inhibited JAK/STAT pathway,and associated proteins uPA,HIF1A and N-cadherin,thereby suppressing the mobility and invasiveness of the HTR-8/SVneo cells,which is conductive to clarify the mechanism of HYXZ in the treatment of early EP.
3.The relationship between fluoride exposure, basal metabolic rate, body fat percentage, and grip strength among adults in rural areas with fluorosis of China
Xuanyin ZHANG ; Meng GUO ; Ruiqin CHEN ; Zichen FENG ; Meng YANG ; Xiaoxue LIU ; Fangfang YU ; Zhiyuan LI ; Yue BA ; Guoyu ZHOU
Chinese Journal of Endemiology 2024;43(1):29-34
Objective:To study the relationship between adult fluoride exposure and grip strength in rural areas of China with fluorosis, as well as the roles of basal metabolic rate (BMR) and body fat percentage (BFP) in the association between fluoride exposure and grip strength.Methods:From April to May 2017, a cluster sampling method was used to conduct a questionnaire survey, physical examination, and biological sample collection on residents aged 18 - 60 in Tongxu County, Kaifeng City, Henan Province (epidemic areas of drinking-water-borne fluorosis). A total of 1 168 subjects were included in the study, including 427 males and 741 females. The fluoride ion selective electrode method and the picric acid method were used to determine the concentrations of urine fluoride and urine creatinine, and the adjusted urine fluoride concentration (CUF) was calculated. BMR and BFP were measured by a bioelectrical impendence method, and the grip strength was measured by a Jamar grip dynamometer. The relationship between CUF, BMR, BFP and grip strength were analyzed using a generalized linear model regression. The mediation effect model was used to assess the mediating effect of BMR and BFP on the association between CUF and grip strength.Results:Female grip strength decreased by 0.28 kg ( P = 0.043) for every 1.00 mg/L increment in CUF. No similar association was found between the two in males ( P = 0.744). Regardless of gender stratification, BMR was positively correlated with grip strength ( P < 0.001). For every 1.00% increase in BFP, female grip strength decreased by 0.18 kg ( P = 0.043). The mediation effect model analysis results showed that the mediation effect ratios of BMR and BFP in the association between CUF and grip strength in female were 65.1% ( P < 0.001) and 8.4% ( P = 0.111), respectively. Conclusion:Fluoride exposure is associated with changes in female grip strength, and BMR changes play a partial mediating role in the association between fluoride exposure and female grip strength.