OBJECTIVE:To establish a method for the dissolution determination of Tamibarotene tablet. METHODS:HPLC was performed on the column of Diamonsil C18 with mobile phase of methanol-water-acetic acid(85∶15∶1,V/V/V)at a flow rate of 1.0 ml/min,detection wavelength was 235 nm,column temperature was 25 ℃,and injection volume was 20 μl. The paddle method was used to determine the dissolution,using pH6.8 phosphate buffer dissolution as medium,medium volume was 900 ml,the rotat-ing speed was 50 r/min,taking samples after 60 min. RESULTS:The linear range of tamibarotene was 0.801-2.804 μg/ml (r=0.999 6);RSDs of precision,stability and reproducibility tests were lower than 3%;recovery was 99.58%-100.90%(RSD=0.25%,n=9);the accumulative dissolution rate of 3 batches of samples was 98.25%,96.54%,92.38%(n=6). CONCLUSIONS:The method simple,accurate with high sensitivity and specificity,and suitable for the dissolution determination of Tamibarotene tablet.

Objective To evaluate the regulatory effect of OX40 co-stimulatory signal on the expression of Foxp3 in inductive regulatory T cells (iTreg) in vitro.Method CD4+ CD25+ naive T cells were isolated from C57BL/6 mouse lymphocyte suspension by MASC CD4+ CD25+ regulatory T cell isolation kit.Inductive Tregs were generated by stimulation of naive T cells in the presence of transforming growth factor beta (TGFβ1),anti-CD3,anti-CD28 and IL-2.The regulatory effect on iTregs was shown by use of OX40 stimulation monoclonal antibody (OX86) or control antibody.Using flow cytometric analysis (FACS),we examined the antibody-based identification of Tregs surface markers CD4 and CD25,along with the intracellular activation marker FoxP3.Results The ratio of CD4+ CD25+ nTregs isolated from mouse lymphatic node was (5.0 ± 0.4)％ vs.(71.8 ± 13.4)％ of TGFβ1-driven iTregs.The ratio of CD4+ CD25+ Tregs was (80.0 ± 1.6) ％ in OX40 stimulation McAb group vs.(86.0 ± 1.4)％ in control antibody group.Furthermore,the expression of Foxp3 was (59.2 ± 0.7) ％ in OX40 stimulation McAb group vs.(70.0 ± 0.8) ％ in control antibody group (P＜0.05).Conclusion TGFβ1-dependent protocol may induce the conversion of naive CD4+ T cells into CD25+ Foxp3+ iTregs.OX40 stimulation can down-regulate the expression of Foxp3 in CD4+ CD25 + iTreg significantly.Thus OX40 molecular may become an attractive target in Tregs-induced transplant tolerance.Further study should be performed to increase the suppressive activity of iTregs through blockade of OX40 signal.

Objective To compare the effect between problem-based learning and lecture-based learning for clinical teaching in the department of cardiovascular medicine. Methods Totally 110 five-year-program cardiovascular interns from June 2011 and June 2012 were selected. They were randomly divided into the PBL group(n=55) and LBL group(n=55). PBL and LBL teach-ing methods were applied in the two groups respectively. Teaching effects were evaluated by exam and questionnaire investigation. SPSS 15.0 was used to do data processing; t test was used to compare the average score of two groups;chi-square test was used to process the results of the questionnaire. P<0.05 signifies sta-tistically significant differences. Results There were statistical differences in examinational average score between PBL group and LBL group ((87.89 ±5.39) vs. (82.63 ±5.26), P<0.05). PBL group had significantly higher satisfaction rate in motivating study interests , deepening understanding of theoretical knowledge, cultivating self-learning ability, training verbal expression and developing clin-ical thinking, etc(P<0.05). Conclusions PBL teaching method demonstrates advantages in teaching of cardiovascular medicine and enhances the teaching effect. But the PBL teaching method should be improved in basic knowledge teaching, cultivation of teachers' ability and case selection.

Objective To estimate the effect of berberine on the proliferation of and expressions of apoptosisrelated factors Bax and Bcl-2 in a human skin squamous cell carcinoma cell line A431.Methods A431 cells were cultured in vitro,and classified into various groups to be treated with berberine at different concentrations (12.5,25,5,100 mg/L) or cisplatin at 250 mg/L (positive control group) for different durations (12,24,48 and 72 hours).The A431 cells remaining untreated served as the negative control group.Subsequently,methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate cell growth,and inverted microscopy to observe cell morphology.Real time quantitative reverse transcription-PCR and an immunofluorescence assay were conducted to measure the mRNA and protein expressions of Bax and Bcl-2 respectively.Statistical analysis was done by multi-way analysis of variance (ANOVA) using the software SPSS 13.0.Results MTr assay showed that berberine inhibited the growth of A431 cells,and the inhibitory effect increased with the increase in concentration (F =1118.312,P ＜ 0.001) and treatment duration (F =510.927,P ＜ 0.001) of berberine.Moreover,there was a significant interaction between the concentration and treatment duration of berberine (F =70.239,P ＜ 0.001).Inverted microscopy revealed that when the concentration of berberine increased,cell density was reduced,and cell morphology changed from polygonal to round with cell body shrinkage.The ratio of bax to Bcl-2 mRNA was elevated with the increase in treatment duration and concentration of berberine,and there were significant differences in the mRNA ratio among cells treated with berberine for different time durations at same concentrations (F =226.231,1300.636,4325.139 for berberine at 25,50 and 100 mg/L respectively,all P＜ 0.001).Immunofluorescence staining indicated that the fluorescence intensity of Bax was enhanced,while that of Bcl-2 was weakened after berberine treatment.Conclusions Berberine inhibits the growth of A431 cells in a dose-and timedependent manner,and may induce the apoptosis of A431 cells via regulating the expressions of Bax and Bcl-2.

Objective To systematically analyze and evaluate the association between the peptidylarginine deiminaseⅣ(PADI4) gene and rheumatoid arthritis (RA) based on the published data,and to provide evidence for the pathogenesis of RA.Methods By selecting five SNPs in PADI4 (rs11203366,rs11203367,rs874881,rs2240340,rs1748033) which had been extensively examined.Meta-analysis on each SNP was performed step by step according to Hugenet manual to investigate the association of the polymorphisms of the PADI4 gene with RA.Results This Meta-analysis enrolled 15 659 RA patients and 22 019 healthy controls from 21 studies worldwide.It demonstrated that rs11203366,rs11203367,rs2240340 and rs 1748033 confered susceptibility to RA in Asian ethnicity (P＜0.01,0.03,＜0.01,＜0.01),while rsl1203366,rsl1203367 and rs874881 confered susceptibility to RA in Caucasian of European ancestry (P=0.0002,0.004,0.03).It also shown that no significant association between rs874881 and RA in the Asian ethnicity populations (P=0.2),or rs2240340,rs1748033 and RA in Caucasian of European ancestry (P=0.18,0.1 ).A linkage disequilibrium study was also performed.The LD study showed that rs11203366,rs11203367,rs874881,rs2240340 and rs1748033 were in linkage disequilibrium both in the Asian ethnicity and Caucasian,which was basically inconsistent with the results of Meta-analysis.The conflicting results should be explained by many aspects such as bias in sample selection,genotyping,and the stratification.Conclusion The PADI4 genotype is partially associated with RA,and the underling mechanisms need further study.Haplotype based research and Metaanalysis would be valuable.

Objective To investigate the value of combined 3D-constructive interference in steady state(3D-CISS) with phase-contrast MR(PC-MR) in diagnosis of hydrocephalus children.Methods We collected retrospectively 28 children with hydrocephalus who caused by obstructive membranes.They all underwent MR imaging examination with a 1.5 T consisting of 3D-CISS, conventional sequences and PC-MR.With the result of surgery and CT cisternography as gold standard,we analysised different sequence in the diagnosis of membranous obstruction.The results of obstructive were analysed by x2 test between 3D-CISS combined with PC-MR and conventional sequences.Results The study included 28 children, of whom 16 showed obstructive membranes in CSF pathways of ventricle and 12 displayed obstructive membranes in CSF pathways of cistern.Conventional sequences diagnosed obstructive membranes of 15 children, and obstructive membranes of 11 children were consistent with gold standard, and 4 cases were false positive.The 3D-CISS diagnosed obstructive membranes of 40, and 12 cases were false positive.There were significant difference between 3D-CISS combined with PC-MR and conventional sequence about obstructive membranes diagnosis of CSF pathways(x2=24.410, P=0.001).Conclusions Making use of jointing 3D-CISS sequence and PC-MR sequence successively, and we can diagnose the position of obstructive membrane for hydrocephalus, which can be helpful to select a perfect surgical approach.

Objective:To evaluate the accuracy of Cerec Bluecam and Cerec Omnicam with 3D printed resin models.Methods:A metal master model including 4 cylindrical metal preparations on the model base,respectively simulating the maxillary canine and the first molar preparations,was prepared.The right canine and the right first molar were respectively marked as A and B,and the left as C and D.The master model was scanned respectively by Cerec Bluecam and Cerec Omnicam for 10 times,10 digital models were genera-ted in Bluecam group and Omnnicam group respectively.Resin models were obtained by a 3D printer based on the digital models,the distance of AB,AC,BD and the diameter(d)of A were respectively measured.Statistical analysis was conducted by SPSS 13.0 soft-ware.Results:The distance of AB,AC,BD of master model and Bluecam group showed significant difference(P <0.05),but d did not(P >0.05);the distance of AC,BD of master model and Omnicam group showed significant difference(P <0.05),but d did not (P >0.05).All the distances of the Bluecam group and Omnicam group showed significant difference(P <0.05).Conclusion:The scanning accuracy of Cerec Bluecam is higher than that of Cerec Omnicam in a single preparation scanning.When the scanning area ex-tending,the scanning accuracy of Cerec Omnicam is higher than that of Cerec Bluecam.

AIM: To explore the neuroprotective action of progesterone(PROG), which has been proved to be a "neuroactive steroid". METHODS: The model of focal cerebral ischemia was established in rats by reverserble inserting a nylon thread with a diameter of 0.2 mm into the anterior cerebral artery through the internal carotid artery. The effect of PROG was assessed by determining water,sodium, potassium, and calcium contents in striatum of rats subjected to 2 h ischemia followed by 22 h reperfusion. RESULTS: The water,sodium,and calcium contents of middle cerebral artery occlusion(MCAO) striatum were obviously higher,the potassium content was obviously lower than those of non-MCAO striatum in I/R and DMSO groups,but there was no significant difference between these two groups.Compared with the result in I/R and DMSO groups , water, sodium and calcium contents significantly decreased, but potassium(P0.05). CONCLUSION: Treatment with PROG can significantly reduce the striatal injury of rats with cerebral ischmia-reperfusion.

Objective:To screen the differentially expressed miRNAs in umbilical cord tissue of children with nonsyndromic cleft lip and/or cleft palate( NSCL/P) using miRNA microarray and comprehensive bioinformatics analysis for the prediction of related the bio-logical process and signaling pathways. Methods:Umbilical cord tissues of 4 cases of healthy newborns' and 4 lip or palate tissues of 4 cases with NSCL/P without other disease aged younger than 2 years were collected. The differentially expressed miRNAs were screened by miRNA microarray. Targets of dysrugulated miRNAs were predicted by TARGETSCAN-VERT, MIRDB and RNA22-HSA. All the gene sets were analyzed by gene ontology and pathway enrichment. Results: MiRNA microarray demonstrated that 254 miRNAs were dysregulated(181 miRNAs were up-regulated and 73 downregulated,P <0. 05). The dysregulated miRNAs targets contained 5029 genes. The dysregulated miRNAs targets were enriched in anatomical structure development,cell adhesion,cell proliferation,cell motili-ty and other biological processes. The dysregulated miRNAs targets were enriched in Wnt, mTOR, cGMP-PKG, TGFβ, PI3K-Akt and other signaling pathways. Conclusion:The target genes set of miRNAs are enriched in multiple biological processes and signaling path-ways related to NSCL/P, which indicate that genetic and environmental factors may influence the development process of NSCL/P.

A matrix solid phase dispersion ( MSPD) method was developed for the simultaneous preparation of samples of 15 mycotoxin biomarkers including deoxynivalenol, aflatoxins and zearalenone from eggs, which were subsequently determined by liquid chromatography-electrospray ionization tandem mass spectrometry ( LC-ESI-MS/MS) under the multiple reaction monitoring ( MRM) mode. For the analysis, the samples were first mixed with C18 particles and loaded into an empty column, then 20 mL of acetonitrile/methanol (1:1, V/V) containing 1 mmol/L ammonium formate was used to elute the sample. The eluent was then dried with nitrogen flow and redissolved into the mobile phase. After filtration, samples were brought into vials and used for analysis. Different from other methods, no extra complicated purification and centrifugation steps were required in the procedure of MSPD. This method had good linearity in the range of 0. 2-100 ng/mL, with the correlation coefficient (r2) greater than 0. 9931. The limits of detection (LODs, S/N=3) and limits of quantification ( LOQs, S/N=3 ) of this method were 0. 05-2 μg/kg and 0. 2-4 μg/kg respectively. Comprehensive extraction recoveries of the 15 compounds ranged from 61% to 90%.