Objective To investigate the effect of metformin on the osteogenic differentiation of human mesenchymal stem cells exposed to PMMA particles. Methods Human placental mesenchymal stem cells were iso-lated and cultured in vitro. The effect of metformin with different concentrations on cell viability was determined by CCK8 assay. The effect of metformin on the mRNA expression of osteogenic genes was detected by using real-time RT-PCR. Calcified nodules were stained by alizarin S. The effect of metformin on the expression of eNOS was also detected by using real-time RT-PCR. Results PMMA particles could inhibit the viability of mesenchymal stem cells. Metformin(0.05 mmol/L)could promote the viability of mesenchymal stem cells exposed to PMMA particles. Metformin(0.05 mmol/L)could increase the expression of osteogenic genes,including OCN,RNUX2,and ALP, in human mesenchymal stem cells exposed to PMMA particles. The calcium deposit was also increased after metfor-min treatment. Results of real-time RT-PCR showed that metformin could increase the expression of eNOS in human mesenchymal stem cells exposed to PMMA particles. Conclusions Metformin can increase the osteogenic differentiation of human mesenchymal stem cells exposed to PMMA particles,partially by inducing eNOS expression.

To investigate the effect of OGT expression inhibited by RNAi on the alteration of tau phosphorylation and glycosylation level in HEK293T cells.The siRNAs targeting OGTgene(OGT-siRNA1～3) were designed and chemically synthesized.The OGT-siRNA1～3 were transfected into HEK293T cells via lipofectamine2000.The efficacy of RNA interference was detected by RT-PCR.The fluorescence of GFP/OGT was counted by fluorescence microscopy after pEGFP/OGT and OGT-siRNA1～3 cotransfected to HEK293T cells for 24 h.The level of tau phosphorylation and glycosylation were detected by Western blot after Plasmid pCI/tau441 and the siRNA cotransfected HEK293T cells for 48 h.OGT-siRNA3(100 nmol/L) could effectively downregulate the expression of OGT gene compared with Mock group(80.0% at mRNA level and 51.3% at protein level).The level of phosphorylation at various sites of the tau was significantly upregulated and the glycosylation was downregulated following the inhibition of OGT gene expression.There is an apparent negative correlation between the modification of phosphorylation and glycosylation of tau protein, the deficient in glucose uptaken/metabolism may be an important pathogenesis of AD.

Arterio-Arterial Fistula ; therapy ; Cardiac Catheterization ; Coronary Vessels ; Female ; Heart Septal Defects, Ventricular ; therapy ; Heart Ventricles ; abnormalities ; Humans

Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; metabolism ; Humans ; Male ; Troponin I ; metabolism

Objective To detect the effect of deep hypothermia on the function of mitochondria in hippocampus after global ischemia in rats and to explore the protection mechanism. Methods The animal model of cardiopulmonary bypass (CPB) was established in rats that were then randomly divided into three groups,ie,control group,normothermia ischemia group and hypothermia ischemia group,eight rats per group.The mitochondria was extracted from the hippocampus of each rats for observing the mitochondrial respiratory function,the activities of succinate dehydrogenase (SDH),the cytochrome oxidese(CCO),the lnembrane fluidity and the content of intramitochondria free calcium and MDA. Resuits The content of intramitochondria free calcium and MDA in the normothermia ischemia group was increased significantly compared to the control group and that in the hypothermia ischemia group wag decreased significantly compared with the normothermia ischemia group(P<0.05).Respiratory state Ⅲ (R3),respiratory state IV(R4),P/O ratio and oxidative phosphorylation (OPR) in the normothermia ischemia group were decreased significantly compared to the control group (P<0.05).R3,R4,P/O ratio and OPR in the hypothermia ischemia group were increased significantly compared with the normothermia ischemia group (P<0.05).Membrane fluidity in the normothermia ischemia group wag decreased significantly compared to the control group (P<0.01),while that in the hypothermia ischemia group was increased significantly compared with the normothermia ischemia group(P<0.05).The activities of SDH and CCO in the normothermia ischemia group were decreased significantly compared to the control group (P<0.01),while those in the hypothermia ischemia group were increased significantly compared with the normothermia ischemia group (P<0.05). Conclusion Profound hypothermia exerts a protective effect on the function of mitochondria in the hippocampus after global ischemia in rats.

Child ; Coronary Aneurysm ; complications ; Humans ; Male ; Mucocutaneous Lymph Node Syndrome ; complications ; Multiple Organ Failure ; complications ; Shock ; complications

Objective To investigate the distribution and antibiotic resistance of isolated pathogens in neonatal sepsis. Methods The results of blood culture and drug susceptibility test in neonates sepsis from January 2012 to June 2013 were retro-spectively analyzed. Results One hundred and thirty-two strains were detected in the blood samples, with 100(75.76%)Gram-positive bacteria, 30 (22.73%) Gram-negative bacteria and 2 (1.52%) fungus. Staphylococcus epidermidis, Escherichia coli and Staphylococcus aureus were the three most common pathogens. Gram-positive cocci was strongly resistant to penicillin (100.00%), erythromycin, selectrin and ampicillin/sulbactam (62.50%-100.00%), but still sensitive to vancomycin and teico-planin. The resistance rate of Gram-negative bacilli to ampicillin was 100.00%, and the resistance rate to cefatriaxone, selectrin and cefuroxime was 61.54%-100.00%. The resistance rate to imipenem and piperacillin/tazobactam was lower. Conclusions The selection of sensitive antibiotics should be based on the pathogens and drug resistance testing for the treatment of neonatal sepsis.

Objective To investigate optimal extraction process of Piper puberulum ( Benth.) Maxim.and qualitative analyze the chemical component of the extracts.Methods Method of solvent heating reflux was used for extraction.On the basis of single factor experiment, L9 (34 ) orthogonal experiment was designed with the variants of extraction frequency, time, material-liquid ratio, and immersion time.Extraction rate as index, extraction processes were optimized to achieve best extraction.The extracts, including total extract, water elution, and ethanol elution, were physiochemically analysed to achieve an initial qualitative result.Results The optimal extraction process was: extractions 3 times for 2 hours, with an 1︰30 material -liquid ratio and 2 hours of immersion, Initial qualitative analyzed the total extracts containing amino acids, polypeptides, proteins, alkaloids, steroids or triterpenes, flavones, saponins, polysaccharides, reducing sugars or glucosides, cumarins, terpene lactones, phenols, and tannins.The water elution containing: amino acids, polypeptides, proteins, saponins, polysaccharides, reducing sugars or glucosides, cumarins, and terpene lactones.The ethanol elution containing: amino acids, polypeptides, proteins, alkaloids, steroids or triterpenes, flavones, polysaccharides, reducing sugars or glucosides, phenols, and tanins.Conclusion The experiments show that optimal extraction process can achieve high extraction yield, stable and practical.

Connexin 43 ; genetics ; Connexins ; genetics ; Heart Defects, Congenital ; genetics ; Homeobox Protein Nkx-2.5 ; Homeodomain Proteins ; genetics ; Humans ; Infant ; Infant, Newborn ; Mutation ; genetics ; Myocardium ; metabolism ; Transcription Factors ; genetics

Objective To investigate the anti-inflammatory mechanism by which curcumin affects TNF-αand IL-8 production in Propionibactierium acnes-induced THP-1 cells. Methods THP-1 cell viability was determined by CCK8 assay. The productions of TNF-α and IL-8 were detected by ELISA assay. Total RNA and proteins were collected for real-time PCR and Western blot analysis, respectively. Results Curcumin didn′t significantly affect the cell viability at 12 h. It decreased Propionibactierium acnes-induced productions of TNF-αand IL-8 in THP-1 cells. Moreover, it decreased TLR2, NF-κB p65, and P-NF-κB p65 expressions in THP-1 cells. Conclusions Curcumin may reduce TNF-α and IL-8 expressions in Propionibactierium acnes-induced THP-1 cells by inhibiting TLR2/NF-κB signaling pathway.