Objective This study aims to understand the characteristics of the time sequence of ICR mouse first mandibular molar tooth germ development through dynamic observation.Methods Tooth germ of Embryos (E11.5,E12.5,E13.5,E14.5,E15.5,E16.5,E17.5 and E18.5) and postnatal (PN1,PN2) mice were obtained.The heads (E11.5-E15.5) and mandibles (E16.5-PN2) of mice were dissected,fixed and embedded for serial sections and HE staining.All the results were assessed under light microscopy.Results The tooth germ underwent various development stages including the bud,cap and bell stages.Mouse odontogenesis was initiated at E11.5.Proliferation of oral epithelium formed the bud stage at E13.5.Then the cap stage was observed at E14.5-E15.5 and the bell stage was appeared beginning from E16.5.The pre-dentin was observed at PN1,as well as the dentin at PN2.Conclusions Establishing the regular development pattern of the first mandibular molar of ICR mice will provide a reliable basis for the future use in the specific tooth germ developmental research.

Objective: To observe the effect of calcium on biological characteristics (proliferation, apoptosis and cell cycle) of ALC ameloblasts. . Methods: ALC cell lines were cultured in vitro in DMEM medium with high glucose at different concentrations (0, 2.0, 2.5, 3.0 and 3.5 mmol/L CaCl2 aqueous solution) for 24 h and 48 h, respectively. Changes of ALC cells under two kinds of incubation time were observed with an inverted microscope. CCK-8 method was used to analyze the effect of calcium ion on ALC cell proliferation. Hoechst staining was used to observe the effect of calcium ion on ALC cell apoptosis. PI staining and FCM method were used to analyze the effect of calcium ions on the growth cycle of ALC cells. Western blot was used to detect the effect of calcium ions on the expression of Cyclin A, Cyclin B and Cyclin D in ALC cells Results: In the 0 mmol/L CaCl2 group, ALC cells were oval or polygonal in shape, and the cells were closely connected and grew like paving stones. In other concentration groups, the morphology of ALC cells did not change significantly after calcium intervention for 24 h and 48 h. Results of CCK-8 method showed that the survival rate of ALC cells slightly decreased with increasing calcium ions concentration after calcium intervention for 24 h and 48 h. However, there was no significant differences in this trend. Results of Hoechst staining showed that the number of ALC cell apoptosis did not increase significantly after different concentrations of calcium intervention for 24 h and 48 h. With the increase of calcium ion concentration, results of PI staining and FCM method showed that the cell cycle of ALC cells gradually increased in S phase and decreased in G1 and G2 phase gradually. Western blot results showed that the expression of Cyclin A and Cyclin B in ALC cells decreased and the expression of Cyclin D increased after different concentrations of calcium intervention for 24 h and 48 h. Conclusion In this study, calcium has no significant effect on the proliferation and apoptosis of ALC cells. Calcium, however, has an effect on the ALC cell cycle. Results of this study show that calcium ions has no obvious toxic or side effects on the ameloblasts, which could be used to explore the possible mechanism and effect of calcium on dental fluorosis.

Objective To explore the impact of broad antigen HLA-Bw4 on disease progression in HIV-1 infected subjects. Methods Three hundred and forty subjects chronically infected with HIV-1 and 69 HIV-1 negative subjects were recruited and HLA-B alleles were typed with sequence-based high resolution typing assay. HLA-Bw genotypes of these HIV-1 infected subjects were determined and their association with CD4+ T cell counts and viral loads were analyzed. Results Sixty-five HLA-B alleles were detected in HIV-1 positive subjects. Subjects with Bw4 (Bw4 homozygotes and Bw4Bw6 heterozygotes ) had higher CD4+ T cell counts ( P = 0. 004 ) and lower plasma viral load ( P = 0.003 ) than subjects without Bw4 ( Bw6 homozygotes). When compared with HIV-1 postive subjects with CD4+ T cell counts above 500 celis/μl, those with CD4+ T cell counts below 500 cells/μl were observed with decreased percentage of Bw4Bw6 heterozygote ( P =0.0002) and increased percentage of Bw6 homozygotes ( P < 0. 0001 ). There is no significant difference in CD4+ T cell counts between Bw4 homozygotes and Bw4Bw6 heterozygote, but lower viral loads were observed in Bw4Bw6 heterozygotes( P = 0. 037 ). Conclusion HLA-Bw4 can confer pretective effects on H1V-1 infected subjects by maintaining higher CD4+ T cell counts and lower viral load, the mechanism behind this effect need further exploration.

Objective To determine whether field triage would reduce median contact-to-device ( C2D ) time in patients with ST-segment elevation acute myocardial infarction ( STEMI ) . Methods Consecutive patients with STEMI underwent primary percutaneous coronary intervention( PCI) from March 2010 to February 2014 in Shanghai Pudong Gongli Hospital were analyzed. Patients were divided into two groups. A total of 121 patients were admitted by field triage and 101 patients by non-field triage. The primary study point was C2D time and the study points secondary included ( door-to-balloor, D2B) time, peak Troponin I ( TnI) levels, hospital mortality and 30 days follow-up mortality. Results Baseline and procedural characteristics between the two groups were comparable. Comparing to non-field triage group, the C2D time was reduced [(92. 0 ± 56. 0)min vs. (131. 0 ± 61. 0)min,P﹤0. 01]. The D2B time was lower in the field triage group vs. the non-field triage group [(55. 0 ±26. 0)min vs. (96. 0 ±31. 0)min,P﹤0. 01]. The percentage of patients with C2D time less than 90 minutes increased significantly from 85. 1% to 98. 3%( P﹤0. 01 ) in the field triage group. Peak TnI level was significantly reduced in the field triage group [(23. 5 ±22. 0) μg/L vs. (43. 5 ± 39. 0) μg/L,P﹤0. 01]. In-hospital mortality and 30 days follow-up mortality did not significantly differ between the 2 groups (3. 3% and 3. 0%, P=0. 885;3. 3% and 5. 0%, P=0. 544, respectively). Conclusions In STEMI patients, field triage was associated with significantly reduced C2D and D2B times.

Objective To investigate how human umbilical cord mesenchymal stem cells (MSCs) in vitro regulate the miRNA profile of activated peripheral blood CD4+T cells from patient with primary Sj?gren's syndrome (pSS). Methods Peripheral blood CD4+T cells from patient with pSS were sorted and divided into healthy naive group, pSS naive group, pSS activated group, MSC treatment group and MSC (pre-stimulated by IFN-γ) treatment group. CD4+ T cells were counted. MiRNA microarray technology was used to detect the expression profile of CD4+T cells, and the expression of miRNA125b and miRNA155 was verified by real time quantification-polymerase chain reaction (RT-PCR). Mean in groups were compared using ANOVA, and multiple comparisons were used with LSD method. Results Both MSCs and IFN-γ-MSCs could inhibit the proliferation of activated CD4+ T cells in a MSC-dependent manner, but there was no significant difference between two groups. Microarray analysis found that the differentially enriched miRNAs in pSS na?ve (vs healthy na?ve), pSS activation (vs pSS na?ve), MSC treatment (vs pSS activation) and pre-IFN-γ MSC treatment (vs pSS activation) were 42 miRNAs, 56 miRNAs, 21 miRNAs and 24 miRNAs, respectively. Furthermore, the expressions of miRNA125b and miRNA155 were verified by RT-PCR and found that miRNA125b relative level in 5 groups was 1.02 ±0.13, 0.80 ±0.11, 0.44 ±0.17, 0.76 ±0.17 and 0.81 ±0.15 (F=18.32, P<0.01), and miRNA155 was 1.5 ±0.8, 3.9 ±1.3, 8.4 ±2.6, 10.1 ±4.2 and 11.2 ±5.0 (F=26.65, P<0.01). Conclusion MSCs can regulate miRNA profile of activated CD4+ T cells in peripheral blood of patient with pSS, and partially reverse down-regulated miR-125b in activated CD4+T cells, which may play a regulatory role in inhibiting the activation of CD4+T cells by MSCs.

Objective To investigate the diagnostic value of texture analysis derived from conventional MR imaging in differentiating benign and malignant breast lesions. Methods Thirty-six patients with malignant breast lesion and 33 patients with benign breast lesion were retrospectively analyzed in our study. All patients underwent conventional MR imaging including axial T1WI, T2WI, and contrast-enhanced T1WI before surgery. Texture features were calculated from manually drawn ROIs by using MaZda software. The feature selection methods included mutual information (MI), Fishers coefficient, classification error probability combined with average correlation coefficients (POE + ACC) and the combination of the above three methods(FPM). These methods were used to identify the most significant texture features in discriminating benign breast lesion from malignant breast lesion. The statistical methods including raw data analysis (RDA), principal component analysis (PCA), linear discriminant analysis (LDA) and nonlinear discriminant analysis (NDA) were used to distinguish malignant breast lesion from benign breast lesion. The results were shown by misclassification rate. Results In the three kinds of sequences, the texture features for differentiating malignant breast lesion and benign breast lesion were mainly from T2WI which had the lowest misclassification rate 4.35%(3/69). The misclassification rates of the feature selection methods were similar in MI, Fisher coefficient and POE+ACC (15.94%to 56.52%for MI;17.39%to 56.52%for Fisher coefficient and 17.39%to 56.52%for POE+ACC). However, the misclassification rate of the combination of the three methods (4.35%to 53.62%for FPM) was lower than that of any other kind of method. In the statistical methods, NDA (4.35% to 27.54%) had lower misclassification rate than RDA (33.33% to 56.52%), PCA (33.33% to 53.62%) and LDA (15.94% to 44.93%). Conclusion Texture analysis of conventional MR imaging can provide reliably objective basis for differentiating benign from malignant breast lesions.

Objective To analyze characteristics of Nef-specific T lymphocyte responses in Chinese HIV-1 recombinant subtype B/C infectors. Methods 19 HIV-1 recombinant subtype B/C infectors infected within 1 year, 40 chronic infectors infected for more than 3 years were enrolled in this cohort study. Elispot assay was used to observe HIV-1 specific T lymphocyte responses in HIV-1 recombinant subtype B/C infectors. Results Nef-specific T lymphocyte responses of interferon-gamma secretion were identified in 15 Chinese HIV-1 recombinant subtype B/C infectors infected within 1 year. The specific T lymphocytes were mainly targeted at four peptides which span from Nef 83 to 135: EVA7081.1, EVAT081.5, EVA7081.6 and EVAT081.48. Responses were identified in 29(75. 2%) infectors with more than 3 years of infection and the specific T lymphocytes were mainly targeted at three peptides which span from Nef 63 to 101 : EVA7081.43, EVA7081.44, EVAT081.45, EVA7081.47, EVA7081.48 and EVA7081.49. The average magnitude of response in infectors with less than 1 year of infection was 284. 13 SFC/106 PBMC. The average magnitude of response in infectors with more than 3 years of infection was 152. 44 SFC/106 PBMC. There was a significant difference between the two groups (U = 91. 000, P = 0. 002). Conclusions HIV-1recombinant subtype B/C infectors at different stages of diseases (less than 1 year and more thank 3 years) can recognize central region of Nef. The magnitude of Nef-specific IFN-γ secretion T lymphocyte responses in this cohort gradually decrease with disease progression.

Objective To analyze the character of Pol-specific T lymphocyte responses and identify immunodominant region recognized in Chinese HIV-1 recombinant subtype B/C infectors at different stages of diseases. Methods Eleven Chinese HIV-1 recombinant subtype B/C infectors infected in 18 months, 25 which infected time more than 3 years and 10 HIV-1-seronegative healthy individuals were enrolled. HIV-1-specific T lymphocyte responses were analyzed by an IFN-γ ELISPOT assay against 249 overlapping peptides spanning HIV-1 Pol protein in the present study. Results Pol-specific T lymphocyte responses of IFN-γsecretion were identified in 8 (72.73%) out of 11 infectors infected in 18 months, the specific T lymphocytes are mainly targe-ted at six peptides which amino acid position from Pol 481 to 631 in reverse transcriptase region: Pol5581, Pol5582, Pol5587, Pol5609, Pol5610 and Pol5615. There was a negative correlation between the breadth of re-sponse and peripheral CD4+ T cell count (P=0.0212, r=-0.762) ; Responses were identified in 15 (60%) out of 25 chronic infectors, the specific T lymphocytes are mainly targeted at four peptides which amino acid po-sition from Pol 241 to 295: Pol5521, Pol5525, Pol5526, Pol5531 and another peptide: Pol5638 which amino acid position from Pol 708 to 722 in reverse transcriptase region. There was a positive correlation between the magnitude of Pol-specific IFN-γ secretion T lymphocyte responses and plasma viremia (P = 0.006 95 , r = 0.660) . None of the seronegative healthy individuals gave the positive responses. Conclusion Chinese HIV-1 recombinant subtype B/C infectors at different stages of diseases mainly recognized different re-gions of Pol.

Objective To analyze character of Nef-specific T lymphocyte responses in Chinese HIV-1 recombinant subtype C/B' and subtype B' infectors and to identify the common immunodominant re-gions recognized by these infectors. Methods Fifty-nine HIV-1 recombinant subtype C/B' infectors and 27 subtype B' infectors were tested by IFN-γ enzyme linked immunospot (ELISPOT) assay using HIV-1 C/B' Nef overlapping peptides. Results Nef-specific T-cell responses of IFN-γ secretion were identified in 44 (74.58%) out of 59 HIV-1 recombinant subtype C/B' infectors. Ten peptides, EVA7081.1,5, 6, 7,43, 44, 45, 47, 48, 49 were mainly recognized. Amino acid position was from Nef63 to 115 and 117 to 139. Twenty of 27 (74.07%) HIV-1 subtype B' infectors recognized peptides. EVA7081.1,2, 43 and 49 were mainly recognized. Amino acid position was from Nef 63 to 77 and 87 to 119. There was no correlation be-tween the Nef-specific IFN-production of HIV-1-specific T cells responses and viral load or CD4 T cell count in both subtype infectors. Conclusion The immunodominant regions, from Nef63 to 77 and 87 to 115 were recognized by both Chinese HIV-1 recombinant subtype C/B' infectors and subtype B' infectors. These re-gions could be used in design of vaccine.

Objective:To evaluate the effect of pinus massoniana bark extract (PMBE) and grape seed extract (GSE) on dentin demineralization caused by acid.Methods:40 root dentin blocks with half of the surface covered were randondy divided into 4 groups (n =10).All samples were subjected to pH cycling for 8 days,and deionized distilled water(DDW),0.1％NaF,12％ PMBE solution and 12％ GSE solution were used as the experimental solutions in the 4 groups.The dentin mineral density(DMD) of the both sides was determined using micro-computed tomography.The D-value of DMD between nn-demineralized and demineralized side (△DMD)was calculated.The samples were observed with field emission scanning electron microscops (FE-SEM).Results:The △DMD of DDW,NaF,PMBE and GSE groups was 198.64 ±59.97,45.94 ±24.21,90.23 ±28.77 and 105.07 ±29.53 respectively.The △DMD between PMBE and GSE groups had no significant difference (P ＞ 0.05),which were both higher than that of NaF group (P ＜ 0.05) and lower than that of DDW group(P ＜ 0.05).The FE-SEM revealed that the dentin tubules in DDW group were completely open,but in NaF group were essentially closed in PMBE group and GSE group were spindle shaped or narrow crack opening.Conclusion:PMBE and GSE had almost the same effect on improving the acid resistance of dentin.