This paper was aimed to investigate the effects of icariin on the expression of p38 mitogen activated protein kinase(MAPK)protein in rat osteoblasts cultured in vitro, to elucidate the signal transduction of icariin in preventing and treating osteoporosis. The results showed that p38MAPK could be activated by icariin within 5min, and Cbfal could also be upregulated, and SB203580, an inhibitor of p38MAPK pathway, could partly inhibit Cbfal upregulation by icariin.

Objective To explore the effect of hypoxia on exosomes secreted by renal tubular epithelial cells and the function of exosomes in chronic kidney diseases.Methods (1) The supernatant of renal tubular epithelial cells which were cultured in normoxia (21％ O2) or hypoxia(1％ O2) for 48 h was collected and centrifuged gradiently to harvest exosomes.Exosomes were identified and compared by transmission electron microscope, nanoparticle tracking analysis, Western blotting and measurement of the protein concentration.(2) Primary peritoneal macrophages of rats were co-cultured with exosomes in different concentrations (1, 10, 50, 100, 300 mg/L).The expression of interleukin-6(IL-6), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS) in cells and supernatant were separately detected by quantitative real-time PCR (qRT-PCR) and ELISA, and the expression of phospho (p)-STAT/STAT and suppressors of cytokine signaling 1 (SOCS1) in macrophages was detected by Western blotting.At last, the expression of inflammatory microRNAs (miR) in exosomes was measured by qRT-PCR.Results (1) The vesicles harvested by gradient centrifugation were less than 150 nm and expressed CD63 which was characteristic of exosomes.Hypoxia had no effect on the morphology of exosomes, but stimulated their secretion.(2) Hypoxic exosomes dose-dependently improved the expression of IL-6, TNF-α, iNOS in macrophages polarized by lipopolysaccharide (LPS) and increased the expression of p-STAT while decreased the expression of SOCS1 (P ＜ 0.01).MicroRNAs referred to inflammation such as miR-155 and miR-27a increased in hypoxic exosomes compared to that in normoxic exosomes (P ＜ 0.05).Conclusions Hypoxia makes exosomes promoted the polarization of macrophages to M1, which may account for the microinflammation in chronic kidney diseases.

Objective To explore the clinical value of real-time three-dimensional echocardiography (RT-3DE) in percutaneous transcatheter closure treatment of atrial septal defect (ASD). Methods The sizes,shapes and sites of ASD were visualized by RT-3DE and measured by full volume rendered three-dimensional echocardiography (3DE) before operation and after closure operation in 11 ASD patients,and compared with those from two-dimensional echocardiography(2DE) and the operation. Results Before the operation,RT-3DE visualization showed that the secundum ASD′s sites of the 11 patients were central and shapes were mostly ellipse. In this study a significant difference was found in the measurements of long distances of ASD between 3DE and 2DE [( 28.9 ? 8.2 )mm vs ( 20.0 ? 7.3 )mm,P 0.05 ]. Maximal diameters of defects measured by 2DE[( 21.1 ? 3.5 )mm,P

Objective To assess the coordinated regulation and the molecular mechanisms of 17β-estradiol and 1,25-dihydroxyvitamin D3 [ 1,25-( OH ) 2 D3 ] on the proliferation and differentiation of osteoblastic MC3T3-E1 cells.Methods MC3T3-E1 cells were cultured in phenol-red free α-MEM medium supplemented with 10％ FBS,MTT assay was performed to determine the effects of 17β-estradiol and 1,25-( OH )2 D3 on MC3T3-E1 cells proliferation.After cells were treated with different agents,cell cycle related genes [ cyclin E,proliferation cell nuclear antigen ( PCNA ),and cyclin-dependent kinase inhibitor 2b ( Cdkn2b ) ] and markers of osteoblastic differentiation [ type Ⅰ collagen ( COL Ⅰ ),alkaline phosphatase ( ALP),osteopontin ( OPN ) ] were detected with SYBR green-based quantitative PCR.ALP activity was detected with BCIP/NBT method.Results 17β-estradiol could promote proliferation of MC3T3-E1 cells,which was accordant to its ability to increase cyclin E and PCNA and to inhibit Cdkn2b mRNA expression in MC3T3-E1 cells.However,1,25-( OH)2D3 had no effect on the proliferation of MC3T3-E1 cells and also did not enhance the proliferation of MC3T3-E1 cells stimulated by 17β-estradiol.On the other hand,17β-estradiol promoted the gene expression of differentiation markers Col Ⅰ,ALP,and OPN,and 1,25-(OH) 2 D3 synergistically increased the expression of these genes with 17 β-estradiol.Conclusion As two of the most important hormones which regulate bone metabolism,estrogen and vitamin D may coordinately promote osteoblast differentiation,but may not regulate osteoblasts proliferation synergistically.

Objective To analyze the pathogen distribution and drug resistance of candida isolated from children with blood infections in our hospital,and to provide reference for clinical effective prevention and treatment.Methods The blood specimens of pediatric patients were collected between January 2009 and December 2015,and were cultured using BacT/ALERT 3D and BD9140 instruments.The candida were separated with Sobaurandps agar culture medium,and identified with chromogenic medium,API 20CAUX test strips or VITEK-2 compact YST card.The minimal inhibitory concentration of 5 drugs were determined by ATB FUNGUS 3 system.Results In 176 cases,92 strains (52.3％) were from neonatal ward,and 46 strains (26.1％) were from PICU.In newborn group,85 strains were isolated from premature,which contained the low and very low birth weight infants (37 strains),pneumonia(20 strains),neonatal respiratory distress syndrome(9 strains).In PICU,the strains were commonly isolated from children with severe infection.Among 176 strains of candida,71 strains (40.3％) were C.albicans,62 strains (35.2％) were C.parapsilosis,16 strains(9.1％) were C.glabrata,9 strains(5.1％) were C.tropicalis,and 18 strains(10.2％) belonged to other candida.Conclusion Candida blood infections can happen at all age of chlidren.The most common strains detected from blood were C.albicans,followed by C.parapsilosis.Most of these strains are susceptible to antifungal drugs,such as fluconazole,except C.glabrata.The sensitive rates to commonly used antifungal drug are more than 93％.The selection of antifungal drugs should be based on the species of strains.

Objective The aim of this study was to explore the effect of low density lipoprotein ( LDL) on the proliferation and differentiation of MC3T3-E1 osteoblasts, as well as the expression of low density lipoprotein receptor-related protein 5(LRP5) and dickkopf-1(DKK1) mRNA of MC3T3-E1 osteoblasts. The possible mechanisms of the treatment of atorvastatin on LDL expression in MC3T3-E1 osteoblasts were also investigated. Methods Proliferation, osteocalcin expression, LRP5, and expression of DKK1 mRNA of MC3T3-E1 with interaction of LDL at 0. 05, 0. 10, 0. 20 mg/ml levels after 24 h, 48 h, 72 h were detected by CCK8, ELISA, and fluorescence quantitative PCR. Furthermore, proliferation, osteocalcin expression, LRP5 and DKK1 mRNA of MC3T3-E1 after the treatment of atorvastatin of 10-6 mol/L and 10-5 mol/L were also be studied, respectively. Results The effect of LDL on proliferation, expression of osteocalcin and expression of LRP5 and DKK1 mRNA in MC3T3-E1 osteoblasts was the most obvious under LDL with 0. 20 mg/ml level. Under that level, atorvastatin (10-6 mol/L or 10-5 mol/L) was able to make the proliferation of MC3T3-E1 osteoblasts in 48 h and 72 h be decreased, while significantly caused upregulation of osteocalcin, LRP5 mRNA expression; and down regulated DKK1 mRNA expression ( all P<0. 05). Conclusions Atorvastatin can reduce the inhibitory effect of LDL on the proliferation and differentiation of osteoblasts. The mechanisms of atorvastatin on osteoblasts are possibly related to the osteoblast proliferation and expression of LRP5 mRNA and DKK1 mRNA of osteoblasts of wnt signal pathway.

Objective To explore the detailed underlying molecular and signaling mechanisms in the effects of icariin on bone formation by an in vitro cell model. Methods The proliferation of MC3T3-E1 osteoblast-like cells was evaluated by MTT, and gene expression of cell cycle related proteins in MC3T3-E1 cells after icariin treatment was detected by real-time PCR. The phosphorylation of MAPK signals, including ERK, P38, and JNK was determined by Western blot, and then the inhibitors of MAPK signals were used to treat cells with icariin alone or together to determine the role of MAPKs in the process of icariin treatment on MC3T3-E1 cell proliferation. Alkaline phosphatase and Alizarin red staining were used to detect the formation of mineralization nodules, and gene expressions of alkaline phosphatase, type Ⅰ collagen, and osteocalcin in osteoblasts after being treated by icariin were evaluated by real-time PCR. ICI182780, and nilutamide was used to decide the participation of estrogen and androgen receptor signals in the process of icariin treatment on the differentiation and mineralization of MC3T3-E1 cells. Results Treatment with icariin promoted MC3T3-E1 cell growth in a time- and dose-dependent manner. This treatment also revealed that icariin increased the expression of mRNAs encoding both cyclin E and PCNA, positive regulators of cell growth, but decreased levels of mRNAs encoding Cdkn2b, a negative regulator of cell cycle progression. When MC3T3-E1 cells were cultured in a differentiated condition, icariin enhanced mineralized nodule formation and increased the expression of mRNAs encoding alkaline phosphatase, type Ⅰ collagen, and osteocalcin. Treatment with icariin significantly induced phosphorylation of both ERK and JNK and this phosphorylated effect occurred very rapidly within 5 minutes and reached peak at 15 minutes. Furthermore, the stimulated effects of icariin on proliferation and gene expression of cyclin E, PCNA, and Cdkn2b in MC3T3-E1 cells were dramatically attenuated by treatment with both U0126 and SP600125, inhibitors of MAPKs. Interestingly, such stimulating effects of icariin were at least partly reduced by treatment with ICI182780, an inhibitor of estrogen receptor. Icariin induced mineralized nodule formation and gene expression of alkaline phosphatase, type Ⅰ collagen, and osteocalcin in MC3T3-E1 cells were also partly reduced when the cells were treated with ICI182780. Conclusions Our findings indicate that the anabolic effect of icariin on bone formation is, at least partly, mediated through the MAPK signaling pathway in order to modulate osteoblast proliferation and differentiation.

Objective To assess the clinical value of high frequency ultrasound diagnosis for little bone fracture of ankle. Methods Thirty-seven ankle wound patients with negative X-ray examination underwent high frequency ultrasound. The second X-ray examination was performed on patients with hinted little bone fracture of ankle, and then CT/MR examination was performed on patients whose second X-ray examination was negative. The consequence of the second X-ray examination was counted and the sonographic features were analyzed. Results ①Little fracture of fibula (lateral malleolus) was in 16, of tibia (medial malleolus) in 12, of talus in 2, of scaphoid bone in 4, of metatarsale in 2 and of calcaneus in one patients. ②The second X-ray examination was positive in 26 patients (26/37, 70.27%), indicating that the detection rate of the second X-ray is higher than that of the first X-ray. ③Sonographic features of little bone fracture of ankle included incomplete bone surface of bone was rough and soft tissue swelling and thickening. No malposition of bone fracture was found. Conclusion High frequency ultrasound is supplementary for X-ray examination, being able to possess important clinical priority in the diagnosis of little bone fracture.

Objective To study the clinical characteristic and antifungal drug sensitivity of neonatal Candida parapsilosis infections in blood and to provide reference for clinical prevention and therapy.Methods The drug resistance of Candida parapsilosis which were isolated from 8 neonatal blood specimen on the laboratory were analyzed and clinical characteristic of infections in neonates were investigated retrospectively.The blood cultured with BD9120.The fungi were isolated with Sabourand Dextrose Agar and CHROMagar colored medium and identified with API20C.The susceptibility test was then performed with FUNGUS3 micro dilution plate.Results Candida parapsilosis susceptibility results to antifungal drugs showed that 5-flucytosine≤ 4 μg/ml,amphotericin B ≤ 0.5 μg/ml,fluconazole ≤ 1 ～ 2.0 μg/ml,itraconazole ≤ 0.125 ～0.125 μg/ml,voriconazole ≤0.06 ～0.06 μg/ml.Seven cases were preterm infants(low birth weight infants or extremely low birth weight infants),one case was term infant after operation of congenital pyloric atresia.Before blood culture,all the 8 cases of Candida parapsilosis sepsis had received broad-spectrum antibiotics and intravenous nutrition.All the 8 cases had received peripherally inserted central catheter,and 3 cases had received mechanical ventilation.Four cases with fluconazole,3 cases with fluconazole and amphotericin B,1 case with fluconazole at the onset but changed VFend,all 8 cases were cured.Conclusion Candida parapsilosis has become the one of the main pathogens of neonatal infection fungal in blood of premature or low birth weight infants,which is sensitive to 5 kinds of antifungal drugs in vitro susceptibility test.Early detection and antifungal therapy can improve the prognosis.