Objective To explore the relationship between the expression of Dishevelled2 and Vangl2 proteins and the development of mouse palate. Methods Twenty-four pregnant mice were randomly divided into eight groups, and the mouse embryos were obtained at eight clock of the pregnant day of thirteen(p13d8h), p13d14h,p13d22h,p14d8h,p14d14h,p14d22h,p15d8h and p15d22h respectively, then paraffin sections were made conventionally.The distrubution and dynamic changes of Dishevelled2 and Vangl2 proteins in the embryonic palatal shelves were detected by immunohistochemistry and image analysis. Results It was found that the two kinds of proteins expressed in the epithelium and mesenchyma of the mouse palatal shelves at different development stages. The expression levels of the Dishevelled2,in both of the epithelium and mesenchyme of the palatal shelves, increased first (p13d8h-p13d22h),then decreased rapidly(p13d22h-p14d14h), and then increased again(p14d14h-p15d22h). The expression of Vangl2 protein in the mesenchyma showed a similar trend to that of the Dishevelled2, but there was no obvious regularity in the epithelium. In addition, the expressive levels of both kinds of proteins in the epithelium were significantly higher than those in mesenchyma of the palatal shelves. Conclusion Dishevelled2 and Vangl2 proteins might directly or indirectly take part in the regulation process of mouse palate morphogenesis.

Objective To study the effects of mitochondrial permeability transition (MPT) on hepatocyte apoptosis and mitochondrial damage,and investigate the effects of curcumin on MPT and the related mechanisms in septic rat.Methods Fifteen healthy male Sprague-Dawley (SD) rats were randomly divided into three groups:sham group,sepsis group and curcumin group,with 5 rats in each group.Septic rat model was reproduced by cecal ligation and puncture (CLP).The rats in sham group were flipped the cecum without perforation and ligation.The rats in the curcumin group were treated with curcumin 100 mg·kg-1·d-1 (dissolved in saline to 10 mL/kg) by oral gavage for 7 days,while the other groups were treated with normal saline.Tissue samples were harvested in each group at 12 hours after operation.Pathological changes in hepatic mitochondria were observed under electron microscopy,concentration of free calcium was examined with confocal laser scanning microscope.After Fluo-3/AM staining,protein and mRNA expression of active caspase-3,Bcl-2 and Bax were examined by Western Blot and reverse transcription-polymerase chain reaction (RT-PCR).Results Under the transmission electron microscope,intact cell membrane,adqulis cytoplasm,and normal and clear mitochondrion was found in the sham group.Mitochondria in sepsis group swelled obviously with mitochondrial cristae broken or disappearance,unclear bilateral membrane structure,while the curcumin group showed much less pathological changes,with few mitochondria swell,and smear bilateral membrane structure.The fluorescence intensity index of sham group,sepsis group and curcumin group was raised successively (417.33 ± 15.88,772.95 ± 42.37,1 560.84 ± 160.78,respectively,F=184.149,P=0.000).The protein and mRNA expression of active caspase-3 and Bax had the highest level in sepsis group,followed by the curcumin group,and that in the sham group was the lowest [active caspase-3 protein (gray scale):1.698 ± 0.061,0.694 ± 0.045,0.246 ± 0.027,F=1 289.667,P=0.000; active caspase-3 mRNA (2-Δ△Ct):1.031 ±0.135,0.578 ± 0.144,0.183 ±0.036,F=66.958,P=0.000; Bax protein (gray scale):1.826 ±0.126,1.254 ±0.140,0.623 ±0.901,F=94.536,P=0.000; Bax mRNA (2-ΔΔCt):2.774 ±0.338,1.661 ±0.226,0.656 ±0.114,F=124.710,P=0.000],all of these values had statistical significance among the three groups (all P＜0.01).While Bcl-2 protein and mRNA had the highest level in curcumin group and lowest level in the sham group [Bcl-2 protein (gray scale):0.716 ±0.091,1.328 ±0.147,1.656 ±0.104,F=84.918,P=0.000; Bcl-2 mRNA (2-△ΔCt):0.617 ±0.118,1.393 ±0.096,1.650 ±0.167,F=83.846,P=0.000].The protein and mRNA expressions of Bcl-2/Bax ratio were lowest in sepsis group,then sham group,and highest in curcumin group [Bcl-2/Bax protein (gray scale):0.726 ± 0.055,1.150 ± 0.043,1.333 ± 0.163,F=46.265,P=0.000; Bcl-2/Bax mRNA (2-Δ△Ct):0.505 ±0.041,0.944±0.097,1.006 ±0.168,F=12.211,P=0.001].Conclusions MPT can lead to mitochondrial dysfunction and further cause hepatocyte apoptosis.Mechanism of effect of curcumin on MPT may be related to reduction of intracellular calcium concentration,promotion of anti-apoptotic Bcl-2 gene expression,inhibition of caspase-3 activation and Bax gene.

ObjectiveTo explore the relationship between the expression of Dishevelled2 and Vangl2 and the embryonic neural tube defects (NTDs) induced by all-trans retinoic acid (RA)in Kunming mouse. Methods Fifty pregnant mice were randomly divided into control and RA-treated groups.RA-treated mice were fed with 30mg/kg RA dissolved with peanut oil on embryo 7.75 days, while the mice of control group were administrated with an equal volume of peanut oil on the same time. Then all the embryos were sampled from pregnant mice at the 4th, 18th, 42nd, 66th and 90th hour after treatment. In situ hybridization and immunohistochemical staining technique were used to detect the expression of Dishevelled2 and Vangl2 in embryonic neural tube. Results The two proteins both existed in the epithelial tissue of the mouse embryonic neural tube and displayed different expression modes at various developmental stages.Compared with the control group, the RA treated group showed a significant decrease (P≤0.05) at the 18th and 42nd hour and a significant increase (P≤0.05) at the 66th hour in Dishevelled2 protein after maternal treatment, and no significant difference was found at the 90th hour. Compared with the control group, the Vangl2 mRNA expression in the RA treated group displayed a significant decrease (P≤0.05) at the 4th and 18th hour and a significant increase (P≤0.05) at the 66th hour after RA treatment, and no difference was found at the 42nd hour. Compared with the control group, the expression of Vangl2 protein in the RA treated group decreased (P≤0.05) at the 18th and 42nd hour, and increased (P≤0.05) at the 90th hour after RA treatment, no difference was found at the 66th hour. Conclusion Excessive RA may interfere with the normal embryonic neural tube closure by regulating the expression of Dishevelled2 and Vangl2.

Objective To investigate the clinical value of neutrophil-to-lymphocyte ratio,platelet-to-lymphocyte ratio and red blood cell distribution width in discriminating disease activity of rheumatoid athritis.Methods Eighty-nine rheumatoid arthritis out-patients of the Affiliated Hospital of Jiangsu University from Jan.2012 to Mar.2013 were recruited into this study.The disease activity index,such as swollen joint count and tender joint count were made by rheumatologists,and laboratory parameters（blood cell analysis,erythrocyte sedimentation rate and C-reactive protein) were measured as well.The enrolled patients were divided into the active RA group and remission RA group based on the DAS28 definition.Differences of NLR,PLR and RDW among groups were analyzed by t test and relations between variables were assessed by Spearman correlation analysis.The t test,non-parametric test and Spearman analysis were used for statistical analysis.Results The levels of NLR (3.2±2.0,2.4±0.9,respectively) and PLR (167±70,133±43,respectively) in active RA group were higher than that of the RA remission group (t=2.28 and 2.50,P=0.02 and 0.01).The level of RDW [(13.9±2.5)％,(13.9±2.4)％,respectively] was not significantly different between the active RA group and the RA remission group (t=0.14,P=0.89).However,compared with healthy controls,the level of RDW [(13.9±2.5)％,(13.2±0.2)％,respectively] was significantly increased in RA patients (t=2.74,P=0.006 9).NLR and PLR were positively correlated with DAS28 (r=0.23,P=0.03;r=0.26,P=0.04,respectively),ESR (r=0.28,P=0.03;r=0.43,P＜0.01,respectively) and CRP (r=0.33,P=0.006;r=0.41,P＜0.01,respectively).However,there was no association between RDW and DAS28 (r=0.01,P=0.93),ESR (r=0.20,P=0.15) and CRP (r=0.05,P=0.71).Areas under curve of receiver operating characteristic (ROC) for assessing disease activity of rheumatoid athritis by NLR and PLR were 0.802 and 0.753 respectively.The cut-off values for discriminating active/remission RA were 2.86 and 143.05,with the sensitivity as 0.66 and 0.63,specificity as 0.72 and 0.65.Conclusion NLR and PLR are significantly associated with the disease activity of RA and can be useful to understand the activity state of the illness.

Objective To observe the protective effect of ulinastatin (UTI) on the intestinal barrier function of septic rats.Methods Septic rat model was established using Sprague-Dawley rats by cecal ligation and puncture (CLP) method.Thirty Sprague-Dawley rats were randomly divided into 3 groups (n =10 for each grop) : sham group, septic group and UTI group.All rats received intraperitoneal injections of 0.9％ saline (10 mL/kg) after and 8 h after surgery.In UTI group, UTI (10 × 104 U/kg in 10 mL/kg saline) was injected after and 8 h after surgery.Collect blood samples after 0, 8, 12 h after surgery to examine levels of procalcitonin (PCT), intestinal fatty acid binding protein (iFABP) and diamine oxidase (DAO) by enzyme-linked immunosorbent assay (ELISA) method.Rats were killed 12 h after surgery to collect intestine tissue samples.Pathological changes of intestine were observed under microscopy, and the expression of tight junction protein-1 (ZO-1) and occludin were analyzed by Western blot.Results In sham group, the mucosa structure was complete and the shape was normal, and villi stood neatly.In septic group, intestinal was expanded, intertinal mucosal was atrophic, villi were scanty.An inflammatory infiltrate with numerous nuetrophils was found in the mucosal.In UTI group, the level of severity was relatively slight.The relative optical density of Western blot images were decreased on ZO-1 and occludin in CLP and UTI groups, and decreased more in CLP group (F =43.15 and 52.23, P ＜ 0.05).At 0h after surgery, the plasma values of PCT, iFABP and DAO were similar in three groups (F =11.17, 22.45 and 13.58, P ＞ 0.05).At 8h and 12h after surgery, values of PCT, iFABP and DAO in septic and UTI groups were much higher than those in the sham group, and those in UTI group were also significantly higher than those in septic group (F8h=85.26, 44.59 and 101.47, F12h =59.44, 49.26 and 69.57, all P＜0.05).PCT, iFABP and DAO levels were first increased and then fell down in sham group, those in septic group were keeping increasing, and those in UTI group were first increased and then kept stable.Conclusion UTI shows protective effect to intestinal barrier function in rats with sepsis.

Purpose The patients with varicose veins of lower limbs need to undergo extensive CT or X-ray examinations with high doses of radiation before and after treatment.The aim of this study is to investigate the value of low-dose CT contrast-enhanced scan for the superficial varicosis of lower limbs.Materials and Methods Forty patients diagnosed as superficial varicosis of lower limbs were selected prospectively.According to the scanning parameters,40 patients were randomly divided into four groups:group Ⅰ (standard group) scanned by 120 kV and 250 mAs,group Ⅱ scanned by 120 kV and 100 mAs,group Ⅲ scanned by 100 kV and 200 mAs,group Ⅳ scanned by 80 kV and 300 mAs.The effective dose (ED),signal noise ratio (SNR),contrast noise ratio (CNR) and image quality were evaluated.Results (① There were no significant differences in age,body mass index and height between the four groups (P＞0.05).② The ED values of the four groups were statistically significant (P＜0.05).③ The differences of SNR and CNR had statistical significance between the four groups (F=3.7 and 3.6,P＜0.05).All images met the diagnostic requirements.Conclusion The lower tube voltage and the higher tube current (80 kV,300 mAs) can reduce the radiation dose and have less influence on the image quality,which can meet the clinical diagnostic requirements and can be used as routine scanning parameters for patients with superficial varicosis of lower limbs.

Objective To analyze the changes in the expression of Toll-like receptors 4 (TLR4) and 7 on the surface of murine bone marrow-derived dendritic cells following dengue virus type 2 (DEN2) infection.Methods DEN2 NGC strain was infected into BALB/c suckling mice through intracranial injection and injected into C6/36 cells to induce the in vivo and in vitro proliferation of DEN2, respectively.RT-PCR was performed to identify DEN2 RNA.Reed-Muench method was used to determine the 50% tissue culture infective dose (TCID50) of DEN2.Dendritic cells (DCs) were prepared by stimulating bone marrow cells isolated form C57BL/6 mice with IL-4 and GM-CSF and then identified by flow cytometry.The prepared murine bone marrow-derived DCs were infected with DEN2 and observed with direct immunofluorescence assay.Dynamic changes in the expression of CD11c, CD86 and I-A/I-E molecules on DCs after DEN2 infection were detected by flow cytometry.Levels of DEN2 RNA and the expression of TLR4 and TLR7 at mRNA level were dynamically detected by real-time quantitative PCR.Results The TCID50 of DEN2 to C6/36 cells was 10-5.8.Murine bone marrow-derived DCs were acquired with a purity of 70% and could be infected with DEN2 in vitro.The percentages of CD86 and I-A/I-E molecules on the surface of DCs infected with 1×105 TCID50 of DEN2 were statistically different from those of the negative control group.Neither of the two groups showed a significant difference in the percentages of membrane molecules over time.However, the percentages of membrane molecules on DCs increased with increasing viral load.Compared with the negative control group, the levels of DEN2 RNA in infection groups were increased with increasing virus load, while the expression of TLR4 and TLR7 on DEN2 infected-DCs at mRNA level was decreased with increasing viral load.Conclusion DEN2 infection promotes the maturation of DCs.Expression of TLR4 and TLR7 on DEN2 infected-DCs at mRNA level decreases with increasing viral load, which suggests that TLR4 and TLR7 are closely related to viral infection and play a certain role in the pathogenesis of DEN infection.

OBJECTIVE:To search a more effective treatment for hepatic functional lesion in primary hepatocellular car?cinoma.METHODS:60patients with primary hepatocellular carcinoma were randomly divided into2groups,treatment group received treatment with polarized solution of Ka and Mg combined with compound glycyrrhizin glycyrrhizinate,while control group underwent treatment with polarized solution of Ka and Mg in combination with decomposed glutathione,both of the treatment courses were2wk～4wk.RESULTS:The decrease in levels of ALT and AST was significantly greater in treatment group,as compared with control group(P

Objective: To investigate the role of Smads signaling pathway in the transdifferentiation of airway epithelial cells into myofibroblasts induced by rhTGF-?1 in vitro.Methods: We exposed cultured 16HBE cells to rhTGF-?1(10?g/L),detected the levels of Phosphorylated Smad2 and Smad3 in nucleus proteins by Western blot at different time points,treated with rhTGF-?1 the 16HBE cells pre-transfected with either the expression plasmid of Smad7(pCDNA3.0/Smad7) or the blank vector(pCDNA3.0),and determined the expression of ?-SMA by immunofluorescent staining and RT-PCR and that of E-cadherin by RT-PCR.Results: The expression of Phosphorylated Smad2 was observed and that of Smad3 increased in the TGF-?1-treated group,with the levels of Phosphorylated Smad2 and Smad3 elevated in a time-dependent manner.The level of ?-SMA was significantly decreased while that of E-cadherin markedly increased in the Smad7 transfection group as compared with the blank vector transfection group 24 h after TGF-?1 stimulation,and no differences were found between the two groups at 36 h.Correspondingly,the number of ?-SMA-positive cells was reduced in the Smad7 group compared with the blank vector group(P

Objective To discuss the effectiveness of implementation of blending learning in teaching of Fundamental of Nursing.Methods 234 nursing undergraduates were divided into the experimental group and the control group.The experimental group adopted blending learning method,while the control group used traditional teaching method.The final term exam and technical test results were compared between two groups.Results Students' practice scores in the experimental group were higher than those of the control group.In the experimental group,about 90.2％ undergraduates accessed learning resources through online teaching platform,82.4％ students considered that their nursing skills improved and 78.4％ students thought that their self-learning capacity strengthened.Conclusions Blending learning gives students a potential time,environment and recourse to learn.By integrating in-class and after-class studying,undergraduates' self-learning ability and nursing skills improved.