OBJECTIVE:Studies have confirmed that basic fibroblast growth factor stimulating periodontal ligament cells can promote the proliferation of human periodontal ligament cells.so as to facilitate the reconstruction of lost periodontal tissues.By use of different concentrations of basic fibroblast growth factor,the in vitro cultured normal human periodontal ligament cells were stimulated to observe the decorin gene expression in the periodontal ligament cells.METHODS:The periodontal ligament cells were isolated and cultured using trypsin digestion method.The third generation of cells at logarithmic growth phase were preserved in DMEM containing 10% DMSO and 20% FBS frozen crvopreservation fluid,then moved into liquid nitrogen preservation at the second day Immunohistochemistry was used to identify anti-vimentin and cytokeratin staining The sixth generation of human periodontal ligament cells were divided into experimental group and control group.The experimental group was cultured in DMEM culture medium containing basic fibroblast growth factor at the concentration of 0.1,1,10 μg/L under standard conditions for 24 hours;control group was cultured with DMEM culture medium under standard conditions for 24 hours.The intracellular decorin gene expression was determined using RT-PCR.RESULTS:By microscopic observation,the cells in the control group had no significant changes,and cells proliferated in the experimental group,before stimulation the cells at the bottom of the bottle became denser than those in sparse regions.Adding basic fibroblast growth factor into periodontal ligament cells,could significantly decrease the mRNA expression of decorin,along with the increase of basic fibroblast growth factor concentration,the inhibition effect gradually weakened and became the strongest in the 0.1 μg/L but the weakest at 10 μg/L.CONCLUSION:The basic fibroblast growth factor stimulation can promote the proliferation of periodontal ligament cells and inhibit the synthesis of decorin in a dose-dependent manner,0 1 μg/L induces the strongest inhibitory effect.

The effect of iodinated contrast on thyroid function and auto-antibodies after a single percutaneous coronary intervention (PCI) was explored.In 96 patients with randomly selected PCI,serum TSH,T3,T4,thyroid peroxidase antibody (TPOAb),thyroglobulin antibody (TgAb) were determined preoperatively as well as by 3-6 months postoperatively.After PCI,the level of TSH,T3,T4,showed no significant changes; TgAb,TPOAb which were positive preoperatively,after 3-6 months were statistically increased(P＜0.05).Iodinated contrast in PCI has no significant influence on thyroid function and auto-antibodies after 3-6 months.

Objective: To investigate the cause of an incident of occupational contact dermatitis in a farm in Tianjin Prefecture, so as to provide insights into occupational safety. Methods: The disinfection process, use of disinfectants and individual protective measures in this farm were collected, and a field epidemiological investigation was conducted to collect the demographic characteristics, history of occupational contact, clinical symptoms, diagnosis and treatment data, and onset of disease among individuals with the same type of job. The cause of this incident was analyzed. Results: There were ten interns exposed to potassium hydrogen sulfate compounds simultaneously in this farm, and then, nine interns developed skin flushing across the body, and swelling and itching of the skin. Among these ten interns, five individuals were admitted to hospitals because of severe symptoms and were then clinically diagnosed as systemic contact dermatitis. All five individuals were cured following treatments. Epidemiological survey showed that all cases had a definite history of occupational contact with potassium hydrogen sulfate compounds but without use of any protective agents. In addition, there were thirty-five individuals with the same type of job in this farm that developed similar symptoms when they joined in the disinfection for the first time, and these individuals were self-cured following cessation to contact; however, recurrence of symptoms was found following contacts again. Conclusion This is a cluster of occupational contact dermatitis caused by exposure to potassium hydrogen sulfate compound.

BACKGROUND:Alendronate is a new generation of diphosphate,which is the second generation of osteoporosis drugs.It is widely used to treat diseases related to increase of bone absorption in clinic.OBJECTIVE:To observe the effect of alendronate on the proliferation and the mRNA expression of osteoprotegerin(OPG) and receptor activator of nuclear factor kappa B ligand(RANKL) of human osteoblasts.DESIGN,TIME AND SETTING:A single sample observational experiment was performed at the Basic Research Institute of Chongqing Medical University from November 2007 to March 2008.MATERIALS:Osteoblasts were isolated from cancellous bone in a surgical operation.Alendronate was the product of Merck Sharp & Dohme S.p.A.METHODS:Osteoblasts were cultured with various concentrations of alendronate(1?10-9,1?10-8,1?10-7,1?10-6,1?10-5,1?10-4 mol/L).Osteoblasts cultured without alendronate were assigned as controls.MAIN OUTC0ME MEASURES:①The morphology and growth of osteoblasts were observed.②Effect of alendronate on the proliferation of osteoblasts were detected by MMT method.③The effect of various concentrations of alendronate on the mRNA expression of OPG and RANKL were detected by using RT-PCR.RESULTS:1?10-5 and 1?10-4 mol/L alendronate inhibited the proliferation of osteoblasts.1?10-9-1?10-6 mol/L alendronate promoted the proliferation of osteoblasts,and the 1?10-8 mol/L alendronate had the strongest effect.Alendronate inhibited RANKL mRNA expression in a concentration-dependent manner,1?10-5 mol/L alendronate had a strongest effect at 72 hours after culture(P

OBJECTIVETo investigate the relationship between the level of di-2-ethylhexyl phthalate (DEHP) and idiopathic oligoasthenospermia by measuring the content of DEHP in the semen samples of different subjects.

METHODSWe obtained semen samples from 100 infertile men with idiopathic oligoasthenospermia, 50 working all the year round in the plastic greenhouse (group A) and the other 50 constantly dining from plastic meal boxes (group B). We also enrolled 50 normal male volunteers as controls (group C). We conducted semen analyses using a computer-assisted sperm analyzer, measured the DEHP concentration by reversed-phase high-pressure liquid chromatography, and subjected the data to statistic processing by t-test and correlation analysis.

RESULTSThe mean concentrations of DEHP in the seminal plasma were (0.72 +/- 0.48), (0.71 +/- 0.49) and (0.21 +/- 0.18) mg/L in groups A, B and C, respectively, significantly higher in A and B than in C (both P < 0.05). The DEHP concentration was negatively correlated with sperm motility (P < 0.05).

CONCLUSIONThe DEHP level in the seminal plasma is higher in infertile men frequently exposed to plastic products than in normal males and excessive DEHP may be one of the important factors of idiopathic male infertility.

Adult ; Case-Control Studies ; Diethylhexyl Phthalate ; adverse effects ; Humans ; Male ; Oligospermia ; etiology ; Plastics ; adverse effects ; Semen ; chemistry

Objective To investigate the distribution of water fluoride and the status of water-improving defluoridation projects,thus to explore the condition of endemic fluorosis in Guangxi Province.Methods According to"The National Technical Scheme for Endemic Disease Contml in 2005",the fuorine content in water Was determined by F-ion selective electrode,children's dental fluorosis was checked by Dean method.and the skeletal fluorosis was checked by the standard of clinical scale of skeletal fluorosis.Results 305 water samples in 61 villages were examined,among which 71 waters were exceeded the standard,accounting for 23.28%(71/305).The projects of defluoriding drinking water were running well except one was discarded.The prevalence of dental fluorosis was 13.55%(356/2627),the prevalence of skeletal fluorosis was 4.02%(65/1615).Conclusions The situation of endemic fluorosis control is not optimistic in Guangxi,which needs fuaher prevention and controls.

Objective To investigate whether macrophage-activating lipopeptide-2 ( MALP-2) in-duces the expression of heme oxygenase-1 ( HO-1 ) in THP-1 cells and its possible mechanism .Methods Human monocyte cells THP-1 were cultured in vitro and then were incubated with various concentrations (0, 0.01, 0.1, 1.0 or 5.0 ng/ml) of MALP-2 for 16 h, or were stimulated by 5.0 ng/ml MALP-2 for different length of time (0 h, 4 h, 8 h, 12 h, 16 h or 24 h).The expression of HO-1 at mRNA and protein levels were detected by real-time PCR analysis and Western blot assay .The enzyme activity of HO-1 was detected by colorimetric analysis.THP-1 cells were pre-incubated with 30 μmol/L of SB203580, PD98059 and SP600125 for 30 min and then were cultured with 5.0 ng/ml MALP-2 for 16 h to investigate the role of mito-gen-activated protein kinases (MAPKs) signaling pathway in HO-1 production.After incubating THP-1 cells with 5.0 ng/ml MALP-2 for different periods of time, NF-E2-related factor 2 (Nrf2) protein was detected by Western blot assay to study the effects of Nrf2 pathway on MALP-2-induced HO-1 expression.Nrf2 and HO-1 proteins were measured by Western blot assay after transfecting THP-1 cells (1×106/well) with Nrf2 siRNA at a final concentration of 100 nmol/L.Results MALP-2 enhanced the expression of HO-1 at mRNA and protein levels as well as the enzyme activity of HO-1 in THP-1 cells in a concentration-dependent manner.The expression of HO-1 protein induced by MALP-2 was significantly inhibited by 30 μmol/L MAPKs specific inhibitors ( SB203580 , PD98059 and SP600125 ) .MALP-2 induced Nrf2 translocation at a concentration of 5.0 ng/ml.The expression of Nrf2 and HO-1 proteins were significantly decreased in Nrf 2 siRNA-transfected THP-1 cells.Conclusion MAPKs and Nrf2 signaling pathways were involved in the MALP-2 induced HO-1 expression .

Objective:To observe the molecular mechanism involved in expression of hemeoxygenase -1 (HO-1) induced by a macrophage-activating lipopeptide-2 (MALP-2).Methods:THP-1 cells were cultured in vitro and stimulated by MALP-2 for 12 h, expression of HO-1 was detected by Western blot .TLR2 and TLR6 neutralizing antibodies incubation , dominant negative plasmids transfection were used to assess the functional of TLR 2,6 in mediating HO-1 expression.Phosphorylation of c-Src and Akt were detec-ted by Western blot, and c-Src siRNA and PI3K inhibitor LY294002 were used to investigate the role of c-Src and PI3K in HO-1 ex-pression.Results:MALP-2 induced c-Src phosphorylation , and TLR2 and TLR6 neutralizing antibodies , or their dominant negatively plasmids could abrogate this effect .In addition, siRNA of c-Src could decrease the phosphorylation level of Akt , and the PI3K inhibi-tor could inhibit HO-1 expression.Conclusion: MALP-2 can induce THP-1 cells expression of HO-1 through TLR2,6/c-Src/PI3K pathways .

Objective To provide experimental evidence for the development of multi-epitope-baseded marker vaccines through investigating the humoral and cellular immune responses in BALB/c mice induced by the multiple antigen peptides (MAPs) with the mimic epitope.Methods Three types of MAPs in eight branched forms containing the mimic epitope of Mycoplasma genitalium adhesion protein (MgPa) were prepared using poly-lysine as the core matrix.The purity of MAPs was analyzed by reverse phase high performance liquid chromatography (RP-HPLC).The molecular weights of MAPs were characterized by Mass Spectrometry.The BALB/c mice were immunized intramuscularly for four times with single or mixed MAPs.The specific IgG antibody and the subtype of IgG antibody in serum of the immunized mice were detected by indirect ELISA.The proliferative responses of the spleen lymphocytes were detected using MTT assay.The ELISA were used to detect IFN-γ and IL-4 levels in the cultured supematant of spleen lymphocytes.Results The three types of MAPs containing the mimic epitopes were successfully prepared with high purity.They,could stimulate mice to produce specific IgG antibodies,of which,the major antibody isotype was Th1 immune response-associated IgG2a.Compared with the single MAP immunization group,the mixed-MAPs immunized mice produced more IgG,IgG1 and IgG2a antibody (P＜0.05).Furthermore,these MAPs could enhance the specific proliferation of spleen lymphocytes in immunized mice and induce the production of IFN-γ and IL-4.The levels of IFN-γand IL-4 in mixed-MAPs group were significantly higher than those of the single MAPs group (P＜0.01).Conclusion The three types of MAPs could induce strong specific cellular and humoral immune responses.The immunological competence of the mixed-MAPs was stronger than those of the single MAP.

Objective Comparative evaluation of flow cytometric immunophenotyping in the diagnosis and differentiation of lymphadenopathy,lymphoma and reactive lymphoid hyperplasia. Methods Ninty-nine fine-needle aspiration specimens from patients with tentative clinical lymphoprofierative disorders were consecutively analyzed by both cytology and flow cytometry with histology results as the gold standard. The three color antibodies including CD3,CD3,CD4,CD5,CD10,CD19,CD20,CD23,CD45,K,λ,FMC7 and CD34 were used for cell composition evaluation and cells with abnormal phenotype. Lymphoma cases were classified according to new WHO classification and subtypes were categorized by immunophenotypic analysis. The results from flow cytometry and cytology were compared. Results By cytological study, 40 of 99 cases were diagnosed with lymphoma, 29 cases were diagnosed with metastatic carcinoma, and 30 cases were diagnosed with reactive lymphoid hyperplasia, necrosis or tuberculosis. Among them, 2 non-Hodgkin lymphoma(NHL) cases were misdiagnosed as reactive lymphoid hyperplasia by cytology. Biopsy was performed in 18 cases of NHL including 16 B-NHL and 2 T-NHL By flow cytometry study, 35 of 99 eases were diagnosed with lymphoma, including 4 cases of lymphoblast lymphoma, 1 case of T-cell lymphoma, and 30 eases of other B-NHL For those 30 cases of B-NHL, 28 cases showed monoclonal light chain expression, and k: λ orλ: k atios exceed 3: 1, and B-cell proportion was (73. 2±27. 2)%. Twenty-six cases could be sub-classified by immunophenotyped. Among 16 histologically confirmed B-NHL cases, only 2 cases diagnosed with follicular lymphoma showed discrepancy with flow cytometry results. In all cases diagnosed with reactive lymphoid hyperplasia and metastasis carcinoma , no abnormal lymphocytes can be found, and k: λ or k: λ ratios were less than 3: 1. Conclusions Fine-needle aspiration analysis with flow eytometrie immunophenotyping can be helpful in diagnosis and differential diagnosis as well as sub-classification of NHL