Objective To evaluate the significance of serum matrix metalloproteinase-3 (MMP-3) and joint ultrasonography in assessing the activity of rheumatoid arthritis (RA) by comparing MMP-3 level and the ultrasonic 7 joints (US7) score in RA patients.Methods Serum MMP-3 level and US7 score were measured in 133 RA patients by immune turbidity and Doppler ultrasound.Synchronous 53 healthy subjects were recruited as controls.Clinical data were collected.Erythrocyte sedimentation rate (ESR),serum level of anti-cyclic citrullinated peptide (CCP) antibody,health assessment questionnaire (HAQ) and disease activity score 28 (DAS28) were measured.The level of disease activity is interpreted as remission(DAS28 ＜2.6),low(DAS 28≥2.6-＜3.2),moderate(DAS 28≥3.2-＜5.1),high(DAS28≥5.1).The discriminating validity of MMP-3 and US7 score in disease was evaluated using receiver operating characteristic (ROC) curve analysis with DAS28 as the reference standard.Results Compared with that in healthy controls [35.20(25.90,48.90) μg/L] and remission patients[33.40(22.60,678.40) μg/L],the MMP-3 level in moderate [105.1 (61.70,172.70) μg/L] and high [363.1 (161.50,475.90) μg/L]groups increased dramatically.US7 score in patients with high disease activity was significantly higher than that in other groups.The level of MMP-3 was significantly correlated with DAS28,HAQ,US7 score,yet did not have correlation with anti-CCP antibody.Serum level of MMP-3 was positively correlated with US7 score (r =0.566,P ＜ 0.001).In evaluating the disease activity,US7 score combined with MMP-3 (AUC 0.863 2) was not superior to MMP-3 alone (AUC 0.854 3),but significantly better than single US7 score (AUC 0.7643,P ＜ 0.05).Conclusions MMP-3 is an effective and simple index in evaluating RA disease activity.The combination of MMP-3 and US7 score does not further improve the efficacy to evaluate disease activity than MMP-3 alone in patients with RA.

Objective To explore the effects of fluoride with different doses on the expressions of TGF-? and Smad2/3 in fibroblast and osteoblast at different periods.Methods Fibroblasts and osteoblasts were exposed to different concentrations of fluoride(0,0.0001,0.001,0.1,1,10 and 20 mg?L~(-1)).The levels of TGF-? protein and Smad2/3 at 2,4,24,48 and 72 h after treatment were measured by using ELISA method.The expression of TGF-? mRNA was tested with RT-PCR method.Results In fibroblasts,the contents of TGF-? protein were decreased in the groups of 0.001,0.1,1,10 and 20 mg?L~(-1) F~-at the time of 2 h and in the groups of 1.0001,0.001,0.1,10 and 20 mg?L~(-1) at the time of 4 h(P

Objective To observe the expression of c-fos mRNA and protein in fluoride treated mouse fibroblast (FB) and osteoblast (OB) and to further explore the effects of c-fos in the osteogenic action of FB. Methods Mouse FB and OB were divided into control group and six fluoride groups (0, 0.0001, 0.0010, 0.1000, 1.0000, 10.0000,20.0000 mg/L F-), and the levels of c-fos protein at 2,4,24,48,72 h and c-fos mRNA at 48 h were measured by using ELISA and RT-PCR methods. Results Compared with the control group, fluoride increased the content of c-fos protein obviously in all FB group(P<0.01); and it is increased in 0.0001,0.0010 mg/L groups at 48 h (0.73±0.04, 0.64±0.14) and 0.0001 mg/L group at 72 h(0.70±0.17) in OB compared with the control group (0.32±0.04,0.27±0.05, P<0.01). Compared with the control group (0.95±0.11), RT-PCR revealed an increasing tendency of the expression of c-fos mRNA at 48 h in FB (1.06±0.16, 1.06±0.12,1.12±0.16,1.04±0.15,1.04±0.10,1.15±0.29), but the difference was not statistically significant(P>0.05); however, a statistically significant difference(P<0.01) of c-fos mRNA in 20.0000 mg/L group(1.40±0.17) in O B was found compared with the control group (1.06±0.06). Conclusion The higher expression of c-fos mRNA and protein in FB induced by fluoride may play an important role in the transformation of osteoblastic phenotype as well as increase the osteogenesis ability in FB.

Objective To examine the expression of ATP-binding cassette transporter A1(ABCA1)in eukaryotie cells and the effect of arsenic resistance after the transfection of eukaryotic expression vector containing ABCA1 gene.Methods HeLa cells were transfected with the recombinant plasmid by lipofectaonmine 2000 (recombinant plasmid group),empty plasmid and untransfected HeLa cell as the control group.The level of the mRNA was examined by real-time PCR,and the expression of ABCA1 protein wag examined by Western blot,the change of cell survival rate was examined by methyl thiazolyl tetrazolium(MTT)after exposure in a series of arsenic [0(contro1),4,8,16,32,64,128 μmol/L]for 48 hours.Results Expression level of ABCA1 mRNA in recombinant plasmid,empty plasmid and untransfeeted groups was(2.09±0.08)×10-4,(0.09±0.02)×10-4,(0.08±0.02)×10-4,there was a significant difference between the groups(F=1499.23,P<0.01).The level of ABCA1 mRNA in recombinant plasmid group was higher than empty plasmid and untransfected group(all P<0.01).Western blot showed that specific protein straps existed at 254×103 in all the three groups,with a similar size to the ABCA1 protein.The amount of the recombinant plasmid group was higher than the other two groups.MTT shows that arsenic concentration at 4,8,16,32,64,128 μmol/L,the survival rates of recombinant plasmid group was(94.8±0.9)%,(86.5 ± 2.6)%, (77.8 ± 2.0)%, (56.0 ± 2.0)%, (23.8 ± 1.7)%, (18.6 ± 0.6)%, higher than that of empty plasmid group[ (85.3 ± 1.1)%, (78.7 ± 0.6)%, (67.8 ± 2.4)%, (43.2 ± 1.5)%, (14.5 ± 1.3)%, (8.0 ± 0.4)%], the difference of survival rate had a statistical signifieance(t = 18.985,6.689,5.922,9.504,9.481,32.634, all P < 0.01). Conclusions ABCA1 protein is over expressed in HeLa cells after transfect ABCA1 gene. ABCA1 protein increases resistance of arsenic in HeLa cells.

Breast Neoplasms ; diagnosis ; Diagnostic Imaging ; methods ; standards ; Female ; Humans

Adolescent ; Adult ; Female ; Humans ; Male ; Occupational Exposure ; Organotin Compounds ; poisoning ; Poisoning ; diagnosis ; therapy ; Retrospective Studies ; Young Adult

Objective To establish the crystal violet plaque assay for detection of virus titer of recombined Tiantan vaccinia AIDS vaccine,and provide more stable method of virus titration for rTV AIDS Vaccine.Methods Optimized the concentration of Vero cells,the time and temperature of virus adsorption,and the time of determination for CPE,then established the crystal violet plaque assay for virus titer of rTV.Counting and analysis the plaques by BioSpot Reader,then analyzed the relativity of plaques counted with BioSpot Reader and manual; Several lots rTV AIDS Vaccine and Tiantan vaccinia were titrated by the method of plaque formation-hemadsorption assay,neutral red and crystal violet plaque assay,then analyzed the relativity of the results of three methods ; meanwhile,the virus titer of samples were determine repeatedly by the crystal violet plaque assay,then calculated the coefficient of variation( CV),and verified the precision of the method; SPSS17.0 was used in statistical analysis of the experimental results.Results When the concentration of Vero cells was 5.0×105-9.O×105 cells/ml,virus been adsorbesd 2 h at 37℃,then cultivated 72 h after adding the culture medium containing methyl cellulose.Plaques counted by BioSpot Reader was highly related with counted by manual (r =0.985),so BioSpot Reader counting can objectively reflect the virus plaques with various size,and reduce the error by manual counting; compared the virus titration for different lots of rTV AIDS vaccine and Tiantan vaccinia with three methods,the crystal violet plaque assay was highly related with plaque formation-hemadsorption assay (r =0.997,P＜0.01 ) and neutral red plaque assay(r=0.980,P＜0.01 ).Conclusion Crystal violet plaque assay was established for virus titration of rTV AIDS Vaccine.

Bone Density ; physiology ; Bone Diseases ; epidemiology ; etiology ; metabolism ; pathology ; Calcium ; metabolism ; China ; epidemiology ; Endemic Diseases ; Fluoride Poisoning ; epidemiology ; etiology ; metabolism ; pathology ; Humans ; Osteoblasts ; physiology ; Oxidative Stress ; Parathyroid Hormone ; metabolism ; Thioredoxins ; metabolism ; Transcription Factor AP-1 ; metabolism

ObjectiveTo investigate the relationship between autoimmune thyroid disorders (AITD) and thyroid cancer (TC) in children.Methods Clinical date from 68 patients who underwent thyroidectomy and were histologically conifrmed with TC were retrospectively analyzed from 2000 to 2008. Age, gender, nodular size, lymph node metastasis, and invasive status of thyroid cancer were compared between patients with and without AITD.Results In 68 patients with TC, 45.6% (31/68) patients had concurrent AITD. Compared with those without AITD, a greater female preponderance, older age, smaller tumour size, higher TSH values, higher TG values, shorter survival time, lower disease free survival rate, and higher relapse rate were found in patients with AITD (P<0.05).Conclusions AITD are associated with TC in children. Follow-up is suggested in children with AITD.

AIM:To investigate the effect of cyclopamine , a Hedgehog ( Hh) signaling pathway inhibitor , on the biological behavior of intrahepatic cholangiocarcinoma cell line RBE .METHODS:The proliferation of RBE cells was detected by cell counting with Typan blue staining and MTT assay , and the apoptosis was analyzed by the flow cytometry . The Transwell invasive cabin assay was used to detect the invasion ability , and Western blot was used to determine the pro-tein expression of Gli 1 and MMP-9 in the RBE cells before and after cyclopamine treatment .RESULTS:Cyclopamine in-hibited the growth of RBE cells in a time-and dose-dependent manner .After cyclopamine treatment for 24 h, 48 h and 72 h, the apoptotic rates were significantly higher than those in control group .In control group , the number of cells invading through the Matrigel of invasion chamber was 154.52 ±13.61, while in experimental group it was 62.00 ±12.17, indica-ting that the invasion ability of the cells declined significantly .Furthermore , Western blot showed that the protein levels of Glil and MMP-9 in the RBE cells were decreased after treatment with cyclopamine for 24 h and 48 h.CONCLUSION:Blockage of the Hh signaling pathway with cyclopamine suppresses the proliferation , promotes the apoptosis and inhibits the invasion ability of RBE cells .