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Objective To investigate the effects of microRNA-155 (miR-155) on immuno-inflammatory reaction of foam cells by targeting MyD88 and the possible mechanism. Methods RAW264.7 macrophages were cultured in vitro and transfected with miR-155 mimics, miR-155 inhibitor, MyD88 siRNA and their negative control respectively, then ox-LDL stimulation was given to build foam cell model. The expression of MyD88 in foam cells was detected by RT-qPCR and Western blot assay. Moreover, the expression levels of interleukin (IL)-10, TGF-β1 and MCP-1 in supernatant were determined by ELISA. Results After being transfected with miR-155 mimics, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The protein level of MyD88 decreased significantly (P<0.05), and the expression levels of IL-10, TGF-β1 and MCP-1 in supernatant also decreased significantly (P<0.05). After being transfected with miR-155 inhibitor, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The expression levels of MyD88 protein and inflammatory cytokines increased significantly (P<0.05). After being transfected with MyD88 siRNA, the expression levels of MyD88 mRNA and protein decreased significantly, and the expression levels of inflammatory cytokines also decreased significantly (P<0.05). Conclusion miR-155 can negatively regulate inflammation by targeting MyD88 through the inhibition of translation.
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AIM To study the interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes.METHODS With maackiain and ephedrine as internal standards,HPLC was adopted in the simultaneous content determination of total tlavonoids (baicalin,wogonoside,baicalein,wogonin and oroxylin A) from Scutellariae Radix and total alkaloids (oxymatrine,oxysophocarpine and matrine)from Sophorae flavescentis Radix in whole prescription component (group) compatibility and single medicinal material part (group) at seven time points (0,15,30,45,60,90 and 120 min),followed by the calculation of their in vitro metabolic rates.RESULTS In whole prescription component (group) compatibility,the contents of baicalin and wogonoside were first increased (0-0.5 h) and then decreased (0.5-2.0 h),those of baicalein,wogonin and oroxylin A showed increasing trends (more obvious within 0.5 h).The stable and gende metabolisms of various alkaloids reached balance within 20 min.CONCLUSION The bioavailability improvement and efficacy enhancement of total flavonoids from Scutellariae Radix may attribute to the compatibility with total alkaloids from Sophorae flavescentis Radix and total polysaccharides from Rehmanniae Radix in Sanwu Huangqin Decoction
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Objective To investigate the effects of microRNA-155 (miR-155) on immuno-inflammatory reaction of foam cells by targeting MyD88 and the possible mechanism. Methods RAW264.7 macrophages were cultured in vitro and transfected with miR-155 mimics, miR-155 inhibitor, MyD88 siRNA and their negative control respectively, then ox-LDL stimulation was given to build foam cell model. The expression of MyD88 in foam cells was detected by RT-qPCR and Western blot assay. Moreover, the expression levels of interleukin (IL)-10, TGF-β1 and MCP-1 in supernatant were determined by ELISA. Results After being transfected with miR-155 mimics, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The protein level of MyD88 decreased significantly (P<0.05), and the expression levels of IL-10, TGF-β1 and MCP-1 in supernatant also decreased significantly (P<0.05). After being transfected with miR-155 inhibitor, the mRNA level of MyD88 remained unchanged compared with that of control group (P>0.05). The expression levels of MyD88 protein and inflammatory cytokines increased significantly (P<0.05). After being transfected with MyD88 siRNA, the expression levels of MyD88 mRNA and protein decreased significantly, and the expression levels of inflammatory cytokines also decreased significantly (P<0.05). Conclusion miR-155 can negatively regulate inflammation by targeting MyD88 through the inhibition of translation.
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AIM To study the interactions of component (group) compatibility of Sanwu Huangqin Decoction in incubation model of rat liver microsomes.METHODS With maackiain and ephedrine as internal standards,HPLC was adopted in the simultaneous content determination of total tlavonoids (baicalin,wogonoside,baicalein,wogonin and oroxylin A) from Scutellariae Radix and total alkaloids (oxymatrine,oxysophocarpine and matrine)from Sophorae flavescentis Radix in whole prescription component (group) compatibility and single medicinal material part (group) at seven time points (0,15,30,45,60,90 and 120 min),followed by the calculation of their in vitro metabolic rates.RESULTS In whole prescription component (group) compatibility,the contents of baicalin and wogonoside were first increased (0-0.5 h) and then decreased (0.5-2.0 h),those of baicalein,wogonin and oroxylin A showed increasing trends (more obvious within 0.5 h).The stable and gende metabolisms of various alkaloids reached balance within 20 min.CONCLUSION The bioavailability improvement and efficacy enhancement of total flavonoids from Scutellariae Radix may attribute to the compatibility with total alkaloids from Sophorae flavescentis Radix and total polysaccharides from Rehmanniae Radix in Sanwu Huangqin Decoction
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Objective To investigate the effect of miRNA-145 (miR-145) on immuno-inflammatory reaction of foam cells by targeting CD40. Methods Mouse macrophage cell line RAW 264.7 cells cultured in vitro were randomly divided into model group (non-transfected), miR-145 mimics group (transfected miR-145 mimics), miR-145 inhibitor group (transfected miR-145 inhibitor) and silencing CD40 sequence group (transfected siCD40). Then oxidized low density lipoprotein (ox-LDL) was used to stimulate for 24 h to establish immune inflammatory damage cell model. Quantitative real-time polymerase chain reaction (RT-qPCR) and Western blot assay were used to detect the levels of CD 40 mRNA and protein of each group. ELISA was used to detect the levels of inflammatory factors interleukin (IL)-1, IL-6 and tumor necrosis factor (TNF) -α in cell supernatant. Results Compared with model group, the levels of CD40 mRNA, CD40 protein and IL-1, IL-6, TNF-αwere all significantly decreased in miR-145 mimics group (P<0.01). After transfected with miR-145 inhibitor, the above indexes were all significantly increased than those of model group and miR-145 mimics group (P<0.01). After transfected with CD40 siRNA, the levels of CD40 mRNA, CD40 protein and IL-1, IL-6, TNF-αwere all obviously decreased compared with those of miR-145 inhibitor group (P<0.01). Conclusion MiR-145 can regulate the immune inflammatory process of foam cells through the target gene CD40, inhibit the activation of CD40/CD40L signaling pathway and inhibit inflammatory response.
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Objective·To evaluate the impact of one-child on body mass index (BMI), and percentage of body fat in primary school students. Methods·All the sample was recruited from 5 elementary schools in Gaohang Town, Shanghai, China. The baseline data of height, body weight, and percentage of body fat was obtained in 2013, and re-measured in 2014 and 2015, respectively. Birth weight, breast feeding, diet and time for physical activities of each children and the highest education level, height, and body weight of their parents were also collected by a self-completed questionnaire. Logistic regression and Mix model was used to analyze the relationship between one-child and BMI, BMI-Z score, and percentage of body fat. Results·A total number of 2515 (1323 boys and 1192 girls) primary school students completed the study and entered the analysis. The percent of one child in this study population was 72.0% (1812/2515). BMI-Z score, time for physical activities, parental education level, and maternal BMI were higher, while the rate of breast feeding was lower in one-child group compared to non-one-child group. The results of Logistic regression showed boys (compared to girls), macrosomia ( ≥4000g vs normal birth weight), overweight father and mother (compared to normal BMI) were risk factors for overweight. The factor of one-child didn't increase the risk of overweight (OR=1.119, 95% CI 0.911-1.374). After potential con-founders adjusted, the annual increase of BMI (β=0.028, 95% CI -0.045-0.100), BMI-Z score (β=0.002, 95%CI -0.034-0.037) and percentage of body fat (β=0.013, 95% CI -0.181-0.207) showed no obvious difference between the two groups. Conclusion·One-child factor showed no obvious relationship with BMI, BMI-Z score and percentage of body fat in primary school students.
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Objective To study the effects of environmental factors,including high temperature,high humidity and low temprtature,on the quality of whole blood.Methods Fresh blood was collected from 9 blood donors and divided into 40 parts.One group that comprised 20 samples was used to assess the effects of storage at high temperature and high humidity (0,3,6,12 and 24 h),and the other group(20 samples)was used to determine the effects of low temprtature(0,1,2 and 3 h).The serum free hemoglobin(FHb)concentration, plasma prothrombin time(PT), activated partial thromboplastin time(APTT),thrombin time(TT)and fibrinogen(FIB)were measured and analyzed.Results The high temperature(27-37℃)and humidity(60%-90%RH)had no effect on FHb, PT, APTT, TT or FIB of whole blood. The low temperature(0-13℃and -18--20℃)had some effect on the quality of whole blood.The FHb of test group was higher than that of control group.There was no significant difference in PT, APTT, TT and FIB between the two groups.Conclusion Under high temperature and humidity conditions, 24 hours of whole blood placement is feasible. Under low temperature or cold conditions,short-term whole blood placement is feasible in a tent with heating facilities.