BACKGROUND:Few reports focus on biocompatibility of nano fluoridated hydroxyapatite for dental implants. OBJECTIVE:To investigate the biocompatibility in vitro of nano fluoridated hydroxyapatite modified for dental implants.
METHODS:We utilized sol-gel method to prepare nano fluoridated hydroxyapatite and nanohydroxyapatite powders. (1) Hemolysis test:0.01, 0.15, 0.2 g/L nano fluorinated hydroxyapatite solution, saline and distil ed water at a volume of 10 mL were added into 0.2 mL diluted rabbit anti-coagulation blood samples, respectively. Then the supernatant was detected by absorbance values. (2) In vitro cytotoxicity test:Passage 2 L929 cel s were respectively cultured in culture media containing 100%, 50%nano fluorinated hydroxyapatite extract, 100%hydroxyapatite extract, phenol solution and RPMI1640. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was employed to measure absorbance values at days 2, 4, 7 of culture.
RESULTS AND CONCLUSION:Hemolysis test in vitro showed the hemolysis rates of nano fluorinated hydroxyapatite groups were less than 5%, which were accorded with haemolysis demand of medical materials. The cytotoxicity test in vitro showed during cultivation, the adherence rate of L929 cel s cultured in 100%and 50%nano fluorinated hydroxyapatite extracts were increasing and cel density was rising up. Cel s were fusiform or polygon, which had no evident differences from negative controls in morphology. Nano fluorinated hydroxyapatite showed nontoxic to L929 cel s in vitro.

Objective To investigate the scale and educational-level composition of nursing education in Guangdong Province,identify the issues in the development process,and provide suggestions accordingly.Methods Data on the scale and education level of nursing education were obtained from the Ministry of Education of China.Results Scale of nursing education at three levels for entering nursing (secondary diploma,advanced diploma,and baccalaureate degree) expanded rapidly during 2006～2010 in Guangdong,with 25.6 thousand recruitments totally in 2010,which as 2.05 times as in 2006.The portion of students recruited in secondary diploma programs had increased gradually,resulted in 83.53％ in 2010 (9.41％ for recruitments in advanced diploma programs and 7.06％ for baccalaureate degree programs).179 and 16 students were recruited in master's and doctoral programs in Guangdong,respectively,during 2006～ 2010.Conclusions The current scale and composition of nursing education in Guangdong should be improved according to the dynamic supply-need relationship of nursing workforce.Initial nursing education should be upgraded by increasing the recruitments of advanced diploma and baccalaureate programs and decreasing the recruitments of secondary diploma programs,expand graduate education,and ensure the quality of education.

Objectives To investigate the epidemiology of high-risk human papillomavirus (HPV) infection and the common genotypes in Liaocheng city, Shandong province, China, and to evaluate the application value of high risk HPV detection in cervical cytology with different pathological conditions.Methods A total of 19 707 permanent female residents in Liaocheng were recruited who were married or had sexual life, aged from 18 to 70 years old.They were screened for cervical cancer by thinprep liquid-based cytology test (TCT) from January 2013 to January 2014.The screen positive rate was 4.24 ％ (837/19 707), and 785 volunteers aged from 21 to 65 years old were recalled.The xMAP bead-based hybridization and flowcytometry analysis were used for genotyping.The data were analyzed by comparison and description.Results According to TCT, among 785 cases, there were 478 cases of atypical squamous epithelium of unknown significance (ASCUS) and atypical glandular epithelium of unknown significance (AGCUS), 175 cases of low-grade squamous intraepithelial lesion (LISL), 127 cases of high-grade squamous intraepithelial lesion (HSIL), 5 cases of squamous cell carcinoma (SCC) and adenocarcinoma (ACC).The positive rate of high-risk HPV was 62.8 ％ (493/785).The risk age of infection was 26-30 years old (87.7 ％, 71/81) and 51-55 years old (79.7 ％, 51/64), while a low risk one was found in patients older than 55 years old (28.6 ％, 14/54).The top five high-risk subtypes of HPV were HPV16 (21.5 ％, 169/785), HPV52 (12.2 ％, 96/785), HPV58 (9.8 ％, 77/785), HPV33 (9.7 ％, 76/785), HPV18 (7.5 ％, 59/785).Single infection accounted for 45.0 ％ (353/785), while multi-infection for 17.8 ％ (140/785).98 cases were infected by two subtypes, 37 cases by three subtypes, 2 cases by four subtypes, 2 cases by five subtypes and 1 case by six subtypes.Conclusions Compared with pure cervical TCT screening, high-risk HPV infection detection is an effective method for cervical cancer screening, which can improve the specificity of cervical cancer screening and reduce the omission diagnostic rate.In Liaocheng, HPV subtypes 16, 52, 58, 33, 18 and multi-infection are more prevalent.Women belonging to 26-30 or 51-55 years old are identified as high-risk population.Screening is important for this group to discover early cervical lesions.

Objective To study the expression and significance of Th17 cells from peripheral blood of patients with rheumatoid arthritis (RA). Methods Intracelluar flow cytomete detection of IL-17/IFN-γ and IL-17/IL-6 was established using anti-CD3/Anti-CD28/IL-23 as stimulators after isolation of untouched human CD4~+T cells from PBMC. There were three groups in the present study: ①healthy controls group; ② RA stable group; ③RA active group. Results The isolation of untouched human CD4~+T cells from PBMC was effective and its purity was over 90%. The percentage of intracelluar IL-17 in CD4~+ T cells from RA patients was increased significantly. Such percentage in active group (1.54±0.41) was higher than that of stable group (0.70±0.21, P<0.01) and both of them were higher than those of healthy controls (0.42±0.12, P<0.01). Under anti-CD3/Anti-CD28/IL-23 stimulation, the percentage of intracelluar IL-17 was also increased significantly(P<0.01). The porcentage of intracellular IFN-γ was similar to that of IL-17, while that of IL-6 was not significantly different. There is an correlation between IL-17 and IFN-γ or IL-6. Conclusion There is an abnormal expression of IL-17 and IFN-γ in human CD4~+T cells in RA patients, which is related to disease activity . Th17 cells may be used as a new marker for the assessment of RA activity.

Objective To investigate the immunoregulatory effects of mesenchymal stem cells (MSCs)on peripheral blood T lymphocytes from patients with systemic lupus erythematosus(SLE)in vitro and their potential mechanism.Methods MSCs were isolated from the bone marrow of 3 healthy human volunteers,cultivated and identified.Under phytohemagglutinin(PHA)stimulating,peripheral blood T lymphocytes from 8 patients with SLE were treated with MSCs with the T lymphocyte/MSC ratio being 50:1 in group B and 5:1 in group C or without MSCs(group A).MTT assay was used to detect the proliferation of T lymphocytes.flow cytometry to analyze the expressions of surface markers CD152 and CD28 on T lymphocytes.and real time PCR to measure the mRNA expressions of interleukin-6 and interferon-γ,in the T lymphocytes.Results MSCs could markedly inhibit the proliferation of T lymphocytcs.The proliferation of T lymphocytes expressed as absorbance value at 570 nm was 0.484±0.032 in group B.0.308±0.025 in group C,significantly lower than that in group A(0.765±0.036,both P＜0.05),and significant difference was also observed between group C and B(P＜0.05).In the case of the percentage of CD28 positive T lymphocytes.group B and C were significantly lower than group A(60.39%±3.92%and 45.05%±3.46%vs 74.73%±3.74%,both P＜0.05),and group B significantly differed from group C(P＜0.05).MSCs had no obvious effect on the expression of CD152 on T lymphocytes,but significantly suppressed the mRNA expression of interleukin-6 and interferon-γ(both P＜0.05).and the suppressive effect was enhanced with the incrgase in MSC count.ConclusionsMSCs exert an immunosuppressive effect on T lymphocytes from patients with SLE,likely through inhibiting the proliferation,CD28 expression,interleukin-6 and interferon-γ mRNA expression of T lymphocytes.

ObjectiveTo investigate the profile of Th17/Treg balance in the peripheral blood of patients with systemic lupus erythematosus (SLE).MethodsThirty-two SLE patients in active disease were selected and 30 SLE patients in remission were included in this study.The control group was consisted of 25healthy individuals.The expressions of IL-17 and FoxP3 on CD4+ T cells in the peripheral blood were assessed by flow cytometry and the mRNA levels of these two cytokines were examined by quantitative PCR respectively.ANOVA was used for statistical analysis.ResultsThe percentage of CD4+IL-17+ T cells and IL-17mRNA expression of the active group were significantly higher than those of the remission group and control group[(l.0l±0.22)％,(2.04±0.63)vs (0.48±0.16)％,(1.12±0.34) vs (0.41±0.12)％,1; P＜0.01].There was no difference between the remission group and control group(P＞0.05).However,the percentage of CI4+FoxP3 + T cells and FoxP3 mRNA expression of the active group were significantly lower than those of the remission group and control group [(2.36±t0.54)％,(0.42±0.16) vs (4.34±0.95)％,(0.87±t0.28) vs (5.09±11.17)％,1; P＜0.01 ],and there was also significant differencesbetween the remission group and control group(P＜0.05).ConclusionThl7/Treg balance shift may exist in the peripheral blood of patients with SLE and the degree of imbalance may be related to disease activity of SLE.

Objective To investigate the change and significance of T follicular helper cells(Tfh) in the peripheral blood of patients with systemic lupus erythematosus (SLE).Methods The percentage of CD4+ CXCR5+ICOS+ Tfh cells and the expression of activation marker CD69 on the Tfh cells of peripheral blood from 60 SLE patients (30 in active and 30 in inactive) and 30 healthy subjects (control group) were determined by flow cytometry.The correlations between the percentage of Tfh cells of SLE patients and SLE disease activity index (SLEDAI),anti-nuclear antibody (ANA) titers and the levels of complement 3 (C3),complement 4 (C4) were analyzed.ANOVA and Pearson's correlation analysis were used for statistical analysis.Results The percentage of the Tfh cells in the peripheral blood of active SLE patients was higher than inactive patients and healthy controls [(0.80±0.17)％ vs (0.63±0.13)％ vs (0.57±0.08)％; P＜0.01],there was also statistical significance between inactive patients and healthy controls (P＜0.05).The expression of CD69 on the Tfh cells in the peripheral blood of active SLE patients was higher than that in inactive and healthy controls [(7.3±1.6)％ vs (5.9±1.3)％ vs (5.2±0.9)％; P＜0.01].There was no statistical significant difference between inactive patients and healthy controls (P＞0.05).The percentage of Tfh cells of SLE patients was significantly related with SLEDAI (r=0.680,P＜0.01) and C3 levels (r=-0.416,P＜0.05),butthere was no correlation between that and the ANA titer,C4 levels (r=-0.042,-0.204,P＞0.05).Conclusion Increased percentage and activity of the Tfh cells in the peripheral blood of patients might contribute to the pathogenesis and development of SLE.

Objective To confirm the clinical diagnosis of complete androgen insensitivity syndrome (CAIS) by molecular genetic testing in a large family. Methods PCR was performed to amplify the coding region of androgen gene, followed by direct sequencing in the patients with CAIS and relatives in this family. Results A missense mutation Arg773His was identified in the patients (homozygous) and carriers(heterozygous). Conclusions Mutation Arg773His in the AR gene leads to CAIS in this family. Molecular genetic testing of CAIS facilitates not only prenatal genetic diagnosis but also preimplantation genetic diagnosis and offers genetic counseling for future pregnancies to abandon the transmission of the mutated X chromosome to the coming generation.

Objective To study the immunoregulatory effects of thalidomide on the peripheral blood T-lymphocytes of rheumatoid arthritis patients.Methods MTr was used to detect the effects of different thalidomide concentrations on the proliferation of T-cells.Flow eytometry was used to analyze T-cells early apoptosis and the T-cells subsets in different concentration of thalidomide.The mRNA expression of IL-6,IL- 10 and TNF-α was measured by RT-PCR method.Results The level of thalidomide at 500 μg/ml inhibited the proliferation of T-ceils and the CD3+CD28+ expression of T-cell subsets,but promoted the early apoptosis and the CD8+CD152+ expression of T-cell subsets.Thalidomine at any concentration could inhibit the mRNA expression of IL-6,TNF-α.However,the level of thalidomide that could promote the mRNA expression of IL- 10 was 100 μg/ml and 500 μg/ml.Conclusion Thalidomide can inhibit the proliferation of T lymphocytes and the expression of CD3+CD28+ on T-cell subsets.It can promote the early apoptosis and the CD8+CD152+ expression of T-cell subsets.Thalidomide inhibits the mRNA expression of IL-6 and TNF-α but promote the mRNA expression of IL-10.Thalidomide has immuno-regulatory effects on rheumatoid arthritis T-cells.

Objective To investigate the immunoregulatory effects of bone marrow-derived mesenehy-real stem ceils (BMSCs) combined with leflunomide (LEF) on mice T-lymphocytes in vitro. Methods BMSCs from BALB/c mice were isolated and expanded. The purity of BMSCs was identified by flow cytometry (FCM). The BALB/c mice's spleen lymphocytes were isolated by using EZ-Sep<'TM> Mouse 1X. Under ConA stimulation, spleen lymphocytes were pretreated with LEF, then washed and co-cuhured with BMSCs. We set up four groups to investigate in this study: group A, spleen lymphocytes alone; group B, spleen lymphocytes with BM- SCs; group C, LEF-pretreated spleen lymphocytes alone and the group D, LEF-pretreated spleen lymphocytes with BMSCs. T-lymphocytes proliferation was assessed by MTT. FCM was used to analysis T-lymphocytes apoptosis and surface markers of CD69 and CD28. The mRNA expression of interleukin (IL)-2, IL-10 were detected by real-time RT-PCR. Results In vitro, the T-lymphocytes'values of A570 nm were significantly lower in group B and group C, compared with group A (group B vs group C vs group A, 0.578±0.042 vs 0.502± 0.040 vs 0.778±0.035, P<0.01), while the value of A<,570 nm>in group D was 0.218±0.033, which was also obviously lower than that in group B and group C (P<0.01). There were no suppressing effects on T-lympho-cytes'activation and expression of IL-2 had been observed. The proportion of apoptotic T-lymphocytes in group B and group D were (2.29±0.32)％ and (4.22±0.98)％, which was significandy lower than that in group A (8.08±1.20) (P<0.01). The expression of IL-10 in group B and C were also lower than that in group A (group B vs group C vs group A, 0.098±0.039 vs 0.054±0.022 vs 1.000, P<0.01 ). Either, the expression of IL-10 in group D was 0.023±0.015, which was obviously lower than that in group B and group C (P<0.01). Conclusion BMSCs combined with LEF show synergistic immunoregulatary effects on mice's T-lymphoeytes.