Abstract:Objective:To investigate the effect of FCCP on differentiation of P19 cells to cardiomyocytes. Methods:P19 cells were treated with 5 umol/L FCCP and untreated cells were used as controls,the morphologic changes of P19 cells at indicated time during the differentiation were observed and photographed under an inverted microscope. The expressions of GATA4,NKX2.5 and cTnT were detected using Real-time PCR and Western blot in the differentiated process. Results:Compared to the control group,the embryoid bodies formed in the FCCP treated group were smaller and were loosely structured. The expressions of GATA4,NKX2.5 and cTnT were observed in the two groups,while FCCP treated cells present with a slower dynamic changes during the differentiation. Western blotting results showed that the protein expressions of GATA4,NKX2.5 in the FCCP treated group on day 5,7 and 9 were lower than that of the control group,respectively. For the expression of GATA4 protein on day 11,there was no significant difference between the two groups,while the expression of NKX2.5 protein in the FCCP treated group was lower compared with the control group. The protein expressions of cTnT on day 5,7 showed no significant difference within the two groups,and with further differentiation,the expression of cTnT protein in the FCCP treated group increased but was still lower than the control group on day 9,11,respectively. Conclusion:FCCP inhibited the differentiation of P19 cells to cardiac myocytes.

Abstract:Objective:To study the potential ability of ABPP on the neuronal differentiation of PC12 cells and to preliminarily investigate the activation of ERK1/2 cascade involved. Methods:By cell culture in the low-concentration of serum,the morphological changes of PC12 cells treated with ABPP at different concentration(0.25,0.50,1.00 μg/ml) were detected. Fluorescent immunocytochemistry was performed to examine the expression of NF-H in differentiated PC12 cells induced by 1.0 μg/ml ABPP. Western blot was performed to detect the acitvities of ERK1/2 in PC12 cells activated by 1.0 μg/ml ABPP for different periods(0 h,6 h,12 h,1 d,2 d,3 d and 7 d) and by different concentrations(0.25,0.50,1.00 μg/ml) of ABPP for 2 days co-cultured with activated(diphosphorylated ERK1/2) antibody and mitogen activated protein kinase(MAP Kinase,MAPK,ERK-1 & ERK-2)antibody. Results:After 3 days treated with ABPP(0.25,0.50,1.00 μg/ml),PC12 cells started to present neuron morphology characteristics. As time went on,the number of the differentiated PC12 cells increased. At the 7th day the differentiated PC12 cells produced the neurite network and at the 14th day the phenomena became more obvious. It showed that ABPP promoted the differentiation of PC12 by in a dose-dependent manner. ABPP activated ERK1/2 in PC12 cells in a dose-dependent manner and a time-dependent manner,with the best concentration of 1.0 μg/ml for 2 days. PD98059 significantly inhibited ABPP-induced phosphorylation of ERK1/2. Conclusion:ABPP could induce the neuronal differentiation of PC12 cells through ERK1/2 pathways.

Abstract:Objective:To study whether increase of O-GlcNAcylation in acute myocardial ischemia causes morphology change of myocardial cells by promoting activity of transcriptional factor CREB. Methods:HEK-293T cells were overexpressed with enzymes OGT and OGA,which regulate O-GlcNAcylation. Total O-GlcNAcylation levels and CREB phosphorylation were detected by Western-blot. Primary cultured rat myocardial cells were treated with drugs DON and PUGNAc to regulate intracellular O-GlcNAcylation levels. Cells morphology was observed by light microscope,and O-GlcNAcylation levels and phosphorylation of CREB were detected by Western-blot. Results:In HEK-293T cells and primary cultured rat myocardial cells,the CREB phosphorylation was positively correlated to the O-GlcNAcylation level. The elevation of O-GlcNAcylation in primary rat myocardial cells reduced the number of cells,but increased sive of the cells. Conclusion:O-GlcNAcylation regulates the activity of CREB positively. The elevated O-GlcNAcylation in acute myocardial ischemia is likely to cause the increase of myocardial cell size and the decease of cell density by promoting the activity of CREB,which may be one of the most important factors leading to myocardial remodeling.

Abstract:Objective:To investigate the underlying mechanisms whereby MDA-MB-231 human breast cancer cell migration is regulated through Wnt5a signaling pathway. Methods:The migration of MDA-MB-231 human breast cancer cells was detected using wound healing assay,and then the migratory ratio of MDA-MB-231 human breast cancer cells blocking the Rac1 expression was measured using siRNA. Results:The activated Rac1 promoted the migration of MDA-MB-231 human breast cancer cells. Blocking of Wnt5a/Dvl2 signaling pathway was capable of retarding the Rac1 activation. Conclusion:We demonstrated that MDA-MB-231 human breast cancer cell migration is regulated by Wnt5a/Dvl2/Rac1 signaling pathway. These findings could provide evidences for designing novel therapy based on inhibition of breast carcinoma metastasis.

Abstract:Objective: To investigate the feasibility of BOP [bis-(2-oxopropyl)-N-nitro samine]-induced Syrian hamsters pancreatic adenocarcinoma model,and explore the character of the modeling process. Preliminary analysis of related biology characteristics provides a controllable,dynamic and comprehensive foundation for BOP induced pancreatic animal model. Methods:Seventy-three syrian golden hamsters were treated with carcinogen,N-nitrosobis (2-oxopropyl) amine weekly for 4 weeks (20 mg/kg,subcutaeous injection),and sacrificed at 2-week intervals after 4-week from the last injection. Results:The experimental group pancreatic tumors began to appear after 20 weeks after first injection,different from previous reports,and tumorigenicity was significant higher in 28 weeks after injection,Primary tumor specimen implantation verified tumorigenicity,parts of experimental results were inconsistent with previous reports. Conclusion:BOP can successfully induce Syrian hamsters poorly differentiated pancreatic adenocarcinoma,and the 28th week after first time injection was the optimally time for sample collection. Some experimental results are worthy of further in-depth study.

Abstract:Objective:To explore inhibition of GLP-1 analogues on prostate cancer cells(LNcap). Methods:LNcap cells were treated with different concentrations(0~100 nmol/L)of exenatide for 24 hours. The Optical Density was measured by CCK-8 analysis .The expression of Bcl-2 and Bax protein were measured by Western blot analysis. Results:The viability of LNcap cells was decreased in a dose-dependent manner after the cells treated with exenatide for 24 hours,as well as the rate of Bcl-2/Bax,and the difference between groups was statistically significant(P < 0.05). Conclusion:Exenatide can induce apoptotic cell death in LNcap cells in a dose-dependent manner。

Abstract:Objective:To study the anticancer effect of nano-As4S4/Mn0.5Zn0.5Fe2O4 particles on human cervical cancer cells through chemotherapy in combination with thermotherapy in alternating magnetic fields. Methods:The nano-As4S4/Mn0.5Zn0.5Fe2O4 particles were prepared by improved technique of chemical co-precipitation and impregnation,and characterized by SEM,EDS and atom fluorescence spectrometer. The heat effect and the release ratio of nano-As4S4/Mn0.5Zn0.5Fe2O4 particles were studied under alternating magnetic fields in vitro. The treatment effect of human cervical cancer cells was detected by MTT assay and FCM. Results:The nano-As4S4/Mn0.5Zn0.5Fe2O4 particles had an average diameter of 20~40 nm with stable release ratio of 13.28% for 48 h,and had very distinct heat effect in alternating magnetic fields and could rise to a steady temperature for tumor hyperthermia(41~46℃). The inhibition ratio of the cells exposed to nano-As4S4/Mn0.5Zn0.5Fe2O4 particles was significantly higher than that exposed to nano-As4S4 and Mn0.5Zn0.5Fe2O4 groups (P < 0.05). The apoptosis rate of cells exposed to nano-As4S4/Mn0.5Zn0.5Fe2O4 particles was much higher than that exposed alone to that exposed to nano-As4S4 and Mn0.5Zn0.5Fe2O4 groups(P < 0.05). Conclusion:The nano-As4S4/Mn0.5Zn0.5Fe2O4 particles can be successfully conducted by improved technique of chemical co-precipitation and impregnation,and the nano-As4S4/Mn0.5Zn0.5Fe2O4 particles combined with magnetic hyperthermia can effectively induce cell apoptosis.

Abstract:Objective:To investigate the protective effect of coenzyme Q10 emulsion on myocardial injury in mice induced by adriamycin and its mechanism. Methods:Coenzyme Q10 capsules and emulsion were administered intragastrically and intravenously to the mice for 5 days,respectively. In the 3rd day adriamycin was given intraperitoneally to establish myocardial injury mice model. After 72h,the level of SOD,CK,MDA and LDH in myocardial cells were tested,as well as the ECG. The level of p53 protein in myocardial cells was assayed by immunohistochemical method cells. Results:Coenzyme Q10 emulsion improved the level of SOD,CK,MDA and LDH in mice myocardial cells,but with no dose-dependency. Also the incidence of arrhythmia was depressed by coenzyme Q10 emulsion. Conclusion:Coenzyme Q10 emulsion has a protective effect on myocardial injury induced by adriamycin,and its mechanism may be related to the antagonism action.

Abstract:Objective:To explore the prognostic significance of NPM1,CEBPA,FLT3-ITD and c-kit gene mutations in cytogenetically normal acute myeloid leukemia (CN-AML). Methods:Bone marrow (BM) samples of 112 patients with CN-AML were collected. Mononuclear cells (MNCs) were extracted and genomic DNA was isolated from BM specimens. Chromosome specimens were prepared using the direct method and short-term culture without phytohemagglutinin and the metaphase chromosomes were banded via improved heat treatment Giemsa R-banding method. Incidence of NPM1,CEBPA,FLT3-ITD and c-kit gene mutations was analyzed using genomic PCR and sequencing. Clinical characteristic,overall survival (OS),disease-free survival (DFS) and cumulative incidence of relapse rate were compared between gene mutant and wild-type patients. Cox regression analysis was performed to estimate the significantly prognostic factors for OS. Results:Incidence of NPM1,CEBPA,FLT3-ITD and c-kit mutation was 21.4%,21.4%,13.4% and 5.4%,respectively. WBC count of the patients with mutational NPM1 was significantly higher than that with wild-type NPM1. Patients with mutant FLT3-ITD or c-kit had significantly lower PLT,higher PB and BM blast proportion than those with wild-type genes. After excluding the patients giving up treatment and undergoing allo-SCT,patients with isolated NPM1 mutations had significantly better OS compared to wild-type patients. Patients with isolated mutated CEBPA had significantly better OS and DFS compared with the wild-type patients. Patients with CEBPA mutation had a significantly lower cumulative relapse rate than the wild-type patients. Cox regression analysis showed that age and BM blast proportion were independent prognostic factors for OS,and CEBPA mutations were associated with OS. Conclusion:FLT3-ITD and c-kit mutations were associated with lower PLT count and higher blast proportion in PB and BM. Isolated NPM1 and CEBPA mutations were associated with better prognosis in CN-AML.

Abstract:Objective:To investigate the expression of FOXC1 in non-small cell lung cancer(NSCLC)and its prognostic value. Methods:The tissue microarrays containing 202 NSCLC tissues and 63 normal lung tissues were constructed. The expression of FOXC1 was detected in these tissues by immunohistochemical method. The relationship between expression of FOXC1 and NSCLC’s clinicopathological and survival parameters were evaluated. Results:The expression of FOXC1(60.87%) in the NSCLC tissues was significantly higher than that in the normal group(25.40%)(P < 0.000 1). In the group of NSCLC,FOXC1 expression was significantly correlated with TNM stage and lymph node metastasis. Kaplan-Meier survival analysis indicated that the survival rate was markedly lower in the patients with high level of FOXC1 than those with low level of FOXC1(P < 0.001).Conclusion:The expression of FOXC1 in NSCLC was significantly correlated with the development and progression of the neoplasms. FOXC1 may play a important role in the evaluation of the prognosis of NSCLC patients.

Abstract:Objective:To investigate the HSP70 and TNF-α levels in thyroid cancer tissues and its relationship with clinicopathological characteristics of patients. Methods:We used RT-PCR and Western blot to detect HSP70 and TNF-α expression in 40 cases of thyroid cancer,20 cases of benign thyroid lesions and 15 cases of normal thyroid tissue and analyzed the relationship between HSP70 gene expression level and thyroid carcinoma pathological features. Results:The differences of the HSP70 mRNA levels among three groups were statistically significant(P < 0.001),and the HSP70 mRNA levels in thyroid carcinomas were significantly higher than that in normal thyroid tissue and benign thyroid lesions(P < 0.05);The HSP70 protein levels in thyroid carcinoma were significantly higher than that in normal thyroid tissue(P < 0.05);HSP70 mRNA expression level was significantly correlated with the thyroid cancer histological grade(rs = 0.810,P < 0.05);The differences of TNF-α mRNA level among three groups were significant(P < 0.05),and further study showed that thyroid carcinoma TNF-α mRNA levels were significantly higher than that in normal thyroid tissue as well as benign thyroid lesions (P < 0.05). Conclusion:Both HSP70 and TNF-α expression were increased in the thyroid carcinoma tissues,and the level of HSP70 expression was correlated with the thyroid cancer histological grade,suggesting that HSP70 plays a critical role in the thyroid cancer developing.

Abstract:Objective:To observe the expression of T helper type 17 (Th17) cells and T regulatory (Treg) cells in the peripheral blood from patients with hepatitis B,and study the relationship between Th17,Treg cells,and clinical parameters. Methods:The frequencies of peripheral Treg and Th17 cells of seventeen chronic hepatitis B(CHB) patients,thirteen acute hepatitis B (AHB) patients and fourteen healthy controls(HC) were determined by flow cytometry. Serum concentration of cytokine interleukin (IL)-17 was detected by enzyme-linked immunosorbent assay(ELISA). The HBV DNA loads were measured by Real-time PCR. The level of serum alanine aminotransferase(ALT) was measured by velocity method. Results:In the periphery of CHB and AHB patients,both Th17 frequency and serum level of IL-17 were significantly increased than those in HC subjects(P < 0.05). The peripheral Treg frequency in CHB patients was significantly higher compared with HC subjects (P < 0.01),whereas there was no significant difference between AHB patients and HC subjects(P > 0.05). Compared with HC subjects,the peripheral Th17/Treg ratio of CHB patients was dramatically lower whereas the ratio in AHB patients were significantly higher (P < 0.05). The peripheral Th17 frequency and Th17/Treg ratio were positively correlated with ALT level whereas Treg frequency was positively correlated with HBV-DNA load. Conclusion:In the periphery of patients with hepatitis B,the expression levels of Th17 and Treg cells were both abnormal,meanwhile,there was a relative Th17/Treg imbalance. Futhermore,the peripheral Th17 frequency,Treg frequency as well as the ratio of Th17/Treg were positively correlated with the level of ALT and HBV DNA load. It is suggested that Th17 and Treg cells may play an important role in the pathogenesis of hepatitis B.

Abstract:Objective:To observe the micro-morphologic of the zirconia surface treated with different roughening methods and test roughness of these surface. Methods:Zirconia blocks were manufactured and divided into four groups receiving different treatments (A,alumina sandblasting;B,tribochemical silica coating;C,hot H2SO4/(NH4)2SO4 etching;D,hot HF/HNO3 etching). An scanning electron microscope (SEM),and an atomic force microscope (AFM) were used to address the treated zirconia surface. The roughness values of the four groups were statistically analyzed. Results:Specimens of group A and B showed similar micro-morphologic,while group C and D presented a better roughening structure. The micron-scale roughness values were ranked as group C>groups A and B> group D (P < 0.05),while the nano-scale roughness value of the two hot acid etching groups were statistically higher than that of the other two groups (P < 0.05). Conclusion:Etching zirconia with hot acids provided a better roughening structure and higher nano-scale roughness.

Abstract:Objective:The purpose of this study was to evaluate the shear bond strengths of heat-pressed and layered ceramics with regard to their corresponding metal alloy. Methods:12 Co-Cr alloy and 12 Au-Pt alloy specimens were cast and divided into 2 groups (n = 6). Metal specimens were veneered with ceramics to produce shear test specimens:experimental groups:PoM (A1:PoM/Co-Cr,A2:PoM/Au-Pt) and two control groups:VMK95 (B1:VMK95/Co-Cr,B2:VMK95/Au-Pt). Shear bond strength testing was conducted in a universal testing machine,and the failure strengths were recorded. SEM was utilized to analyze the metal-porcelain interface. Results:Bond strength of PoM groups was significantly higher than that of VMK95 groups and all the groups displayed mixed failure mode. SEM revealed morphologically that in the PoM groups,the veneer layer contacted more closely and tightly to the alloy,and fewer pores were shown in the interface. Conclusion:Heat- pressed Technique shows greater bond strength between metal and the veneering porcelain.

Abstract:Objective:To explore the microstructure of the bonding interface between the fiber post and root canal wall,which was processed with different bonding material and methods. Methods:Fifteen sound canines were root filled after sectioned transversely 2mm coronal to the labial cemento-enamel junction. Fiber post was placed with three different root canal adhesive material(n = 5):RelyX Unicem(3M),DC core(Kuraray),Luxacore Z. Each root specimen was cross sectioned across into three samples (+/- 2.0 mm in thickness). In each group 3 samples were chosen to observe under Scanning Electron Microscopy(SEM). Results:SEM observation of the ashesive interface specimens showed that different bonding treatment caused to form varying degrees of hybrid layer and resin tags. Conclusion:SEM observation of the ashesive interface specimens reveals that the mixed layer and resin tags are the key factors in approaching better bond.

Abstract:Objective:To analyze the correlation of ILI and positive rate of nucleic acid based on data of influenza surveillance in Jiangsu Province. Methods:The ILI number and positive rate of nucleic acid detection among age groups of three years in Jiangsu Province were caculated and the correlation of ILI and the positive rate was analyzed. Results:From 2011 to the 30th week of 2013,there were differences between ILI and positive rate of nucleic acid detection among age groups.The correlation of ILI and the positive rate of nucleic acid detection was not statistically significant between age of 0~4 years and 5~14 years,although it was significantly different among age groups >14 years. Conclusion:We can achieve more scientific and effective influenza surveillance by improving the quality of ILI reporting and specimen collection and adjustment the monitoring program.