ABSTRACT
Aim To study proliferation capacity of cell and the target relationship between microRNA and Runx2 after effect of puerarin on osteoblasts MC3 T3-E1 . Methods The proliferation capacity of cell was detected by MTT after effect of puerarin on osteoblasts MC3 T3-E1 . The vitality of osteoblasts was detected by activity of alkaline phosphatase. The expression level of mRNA and protein of Runx2 were detected by real-time quantitative PCR and Western blot. The result of miRNA expression spectrum was compared with the predicted result to determine the Runx2-targeting miR-NAs. The expression levels of miRNAs possiby targeted to Runx2 were detected by real-time quantitative PCR. The RhoE 3′UTR vector and RhoE mut 3′UTR vector were constructed. miRNA-204 mimics and miRNA-204 NC were synthetised. The target genes were verified by dual luciferase report gene assay. Results After osteo-blasts treated with puerarin, proliferation capacity and activity of cells were enhanced , expression levels of mRNA and protein of Runx2 were both increased , the expression levels of miRNA-204 and miRNA-344 f-5 p were declined, the expression levels of miRNA-2861 was increased,the expression levels of miRNA-23a-5p, miRNA-770-5 p and miRNA-871-5 p showed no obvious change. According to the results of dual luciferase re-porter gene method after cell transfection of 48 h, only set of 3′UTR Runx2+mimics the miRNA-204 of fluo-rescein protein expression level decreased significantly, showing only the miRNA-204 inhibits Runx2 3′UTR report gene expression. Conclusion Puerarin pro-motes the proliferation of osteoblasts and regulates the miRNAs which possibly target to Runx2 .
ABSTRACT
Objective To investigate the relationship and clinical significance between the concentration of 25-hydroxy vitamin D3 [25-(OH)D3 ]in the serum and community-acquired pneumonia(CAP)in infants.Methods The study recruited 98 infants.There were 68 cases of mild pneumonia and 30 cases of severe pneumonia.The con-centration of 25-(OH)D3 in serum,IgA and alkaline phosphatase(AKP)were measured.33 infants who accepted physical examination in the outpatient department were selected as control group at the same time.Results The serum 25-(OH)D3 concentration of severe pneumonia was (21.09 ±7.32)ng/mL,mild pneumonia was (25.77 ± 8.06)ng/mL and the control group was (37.13 ±6.94)ng/mL.The differences among the three groups had statisti-cally significant differences(t =9.18,3.72,5.34,all P 0.05).The prevalence of Vitamin D deficiency was 40.0%,which was higher than mild pneumonia patients(20.6%)and the healthy children(9.1%)(χ2 =8.43,17.55,all P <0.05).Conclusion The serum 25-(OH)D3 concentration and IgA of CAP patients were lower than healthy children,especially in severe pneumonia cases.The prevalence of Vita-min D deficiency was significantly higher than healthy children.There were no relevance between the serum concentra-tion of 25-(OH)D3 ,IgA and AKP.25-(OH)D3 plays an important role in the development of CAP.The low level of serum 25-(OH)D3 concentration may be one of the risk factors and has correlation to the severity of pneumonia.