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 investigate the immunoregulatory effects of mesenchymal stem cells (MSCs) on the peripheral T-lymphocytes of lupus nephritis in vitro. Methods MSCs were isolated and expanded from human bone marrow cells. The purity of MSCs was identified by flow cytometry (FCM). The MSCs (4×104, 1×104, 2×103) were added into wells containing peripheral blood lymphocytes (2×105) from lupus nephritis in the presence of phytohemagglutinin [PHA). Samples were divided into the following groups: group A:T-lymphocytes alone; group B: MSCsl with T-lymphocytes(MSCsl:T=1:5); group C: MSCs2 with T-lymphocytes (MSCs2:T=1:20); group D: MSCs3 with T-lymphocytes (MSCs3:T=1:100). The proliferation of T-lymphocytes was assessed by MTT. FCM was used to analyze the apoptosis of T-lymphocytes and surface markers of CD28 and CD152. The gene expression of interferon γ (IFN-γ), interleukin-10 (IL-10), transforming growth factor-β1 (TGF-β1) were detected by real-time RT-PCR. Results In vitro, with the presence of MSCs, peripheral blood T-lymphocytes from lupus nephritis were statistically significantly decr-eased in their proliferative activities , apoptosis and CD28 expression in a dose-dependent manner. No inhibitory effects on CD152 expression of T-lymphocytes had been observed . MSCs promoted the gene expression of TGF-β1 and inhibited the gene expression of IL-10, IFN-γ. Conclusion MSCs can inhibit the proliferative activities, apoptosis and CD28 expression of peripheral blood T-lymphocytes of lupus nephritis, increase gene expression of TGF-β1 and lower the gene expression of IL-10, IFN-γ, which may play an important role in it's immunosuppressive effects on lupus nephritis.

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 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 investigate the immunoregulatory effect of thalidomide on the peripheral blood T-iymphocytes in systemic lupus erythematosus patients in vitro. Methods T-lymphoeytes were treated by thalidomide with different concentrations, then the proliferation of these T-lymphocytes proliferation was deteted by MTT while apoptosis and lymphocyte activation marker were analyzed by flow cytometry. The mRNA expression of IL-6, IL-10 and TNF-α was measured by real-time RT-PCR, One-way ANOVA was used for statistical analysis. Results In vitro, thalidomide inhibited the expression of CD3~+CD28~+ [500 μg/ml group vs the control group, (48±9)% vs (57±9)% P<0.05]. The pro-portion of apoptotic T-lymphoeytes in the 500 μg/ml group was (36±8)%, which was significantly higher than that in the control group [(23±5)%,P<0.05 ]. The values of A_(570nm) T-lymphocytes were significantly lower in the 100 μg/ml group, 300 μg/ml groupand 500 μg/ml group compared with those of the control group ( 100 μg/ml group vs 300 μg/ml group vs 500μg/ml group vs the control group, 0.39±0.05 vs 0.34±0.04 vs 0.30±0.03 vs 0.51±0.07, P<0.05), while thalidomide promoted the expression of CD8~+CD152~+ [ 100 μg/ml group vs 500 μg/ml group vs the control group, (5.0±0.6)% vs (7.8±0.7)% vs (4.2±0.6)%, P<0.05 ]. 500 μg/ml thalidomide inhibited the mRNA expression of IL-6, 2.5～500 μg/ml thalidomide inhibited IL-10, TNF-α mRNA expression of T-lymphocytes.Conclusion Thalidomide can inhibit the proliferative activities and CD28 expression of T-lymphocytes,reduce mRNA expression of IL-6, IL-10, TNF-α, stimulate CD28 expression and apoptosis of T-lymphocytes. These effects may play an important role in it's immune-suppressive effects on systemic lupus erythematosus.

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.

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.