0.05);while great significances were found between group C and group B and A(P

Objective To investigate the change of biological features of macrophages after transfected by Toxoplasma gondii GRA 1 genes (P24). Methods The transfected cells Cyto P24 RAW264.7, Nuc P24 RAW264.7, Mito P24 RAW264.7 and ER P24 RAW264.7 were studied by RT PCR to determine the P24 mRNA expression. Growth features of the cells were examined with microscopy and the cell growth curve was developed. Results In four cell lines, expression of ER P24 RAW 264.7 was found to be higher than the other three, and there was no P24 mRNA expression in either of the cells without P24 insert. The attachment and the proliferation of ER-P24-RAW264.7 were more rapid than normal RAW264.7. Conclusion Transfection of mouse macrophages ER RAW264.7 strain with T. gondii P24 gene leads to a prominent change of biological features in the studied cell line.

Objective To investigate the signaling role of arachidonic acid in the invasion of RAW264.7 macrophage by Toxoplasma gondii . Methods Rate of infection was calculated by both light microscope and flow cytometer. Fluorescent emission spectra were recorded using a microspectrofluometer for the concentration of cytoplasmic free calcium. Results Calcium concentration in macrophages and rate of infection increased with a higher concentration of exogenous arachidonic acid in a dose-dependent manner. The invasion was dependent on the mobilization of calcium from the extracellular medium and from intracellular stores and followed the influx of calcium into the parasitized cell. Conclusion Arachidonic acid may enhance the rate of infection via calcium transduction pathway.

Objective To investigate the signal transduction pathway of arachidonic acid(AA) and prostaglandin E-2(PGE-2) synthesis in macrophage invaded by Toxoplasma gondii. Methods Synthesis of AA and PGE-2, expression of COX_2 mRNA and protein following stimulation infection by Toxoplasma gondii were evaluated in RAW264^7 cells by ELISA, RT_PCR and Western blotting after treatment with calcium channel blocker verapamil, chelator of extracellular calcium EGTA and inhibitor of CaM trifluoperazine (TFP), selective PKC inhibitor H7. Results Production of AA and PGE-2 induced by tachyzoite was significantly inhibited by EGTA, TFP and BAPTA/AM, and the PGE-2 production was inhibited by H7, with a reduced expression of COX_2 mRNA and protein in a dose_dependent manner. Conclusion The parasite down_regulates macrophage functions by affecting PKC signaling pathways, and triggers a biochemical cascade whose signals ultimately conduct to the secretion of immunosuppressive molecules PGE-2.

Objective To explore prostaglandin E 2 (PGE 2) production pathway in Toxoplasma gondii-infected macrophage RAW264.7 cell line.Methods Cells were incubated with Toxoplasma gondii tachyzoites and lipopolysaccharides (LPS). Prostaglandin synthesis and arachidonic acid in supernants were detected with ELISA and gas chromatogram. Expression of cyclooxygenase-1/cyclooxygenase-2 (COX-1/COX-2) mRNA and protein following stimulation with LPS or infection of Toxoplasma gondii were evaluated with RT-PCR and Western blot in presence or absence of peculiar antagonists of PGE 2 production. Results PGE 2 synthesis of macrophages began at 4-8 h after invasion with Toxoplasma gondii and saturated at 12-16 h. Expression of COX-2 mRNA peaked at 4-8 h, and diminished in presence of both indomethacin and nimesulide, COX-2 protein expression was not affected by them. Expression of COX-1 mRNA and protein were constant and not affected by either indomethacin or nimesulide. Conclusion Toxoplasma gondii may induce macrophages prostaglandin E 2 synthesis via cyclooxygenase-2 pathway.

ObjectiveTo explore the effect of TLR4,PI3K and NF-κB on the migration of asthmatic airway smooth muscle cell(ASMCs) induced by airway epithelial cells.MethodsPrimary ASMCs were cultured by the method of cell digestion.Cell culture supernatant of RTE cells were collected by TNF-α stimulation of epithelial cells.Detected the IL-8 and RANTES levels in the supernatant.The effect of TLR4/PI3K-related signaling molecules on the migration of asthmatic ASMCs induced by epithelial cells were detected by Modified Boyden chemotaxis chamber with anti-TLR4 antibody,Wortmannin and PDTC drugs as a tool.ResultsThe levels of IL-8 and RANTES in the supernatant of TNF-αgroups were significantly increased,and that in the 20 ng/ml group was significantly higher than other groups ( P＜0.01 ).Compared with control group,the transmembrane migration of asthmatic ASMCs from other groups was significantly increased (P＜0.01 ).The transmembrane migration of asthmatic ASMCs from treated groups was significantly increased than asthma group (P＜0.01).The migration of asthmatic ASMCs from TNF-α+anti-TLR4 antibody group,TNF-α+Wortmannin group and TNF-α+Wortmannin+PDTC were significantly decreased than that of TNF-αgroup( P＜0.01 ).The migration of asthmatic ASMCs from TNF-α+Wortmannin+PDTC were significantly decreased than that of TNF-α+Wortmannin group (P＜0.05).ConclusionTLR4/PI3K-related signaling molecules involved in the transmembrane migration of asthmatic ASMCs induced by airway epithelial cells and may be one of the mechanisms of airway remodeling of asthma.

Objective To indentify the cognitive status of Chinese patients to acne and the influencing factors to theirs' cognitive status,so as to provide solid evidences for the prevention and treatment of acne.Methods A self-designed questionnaire was made to conduct this survey of 16,156 acne patients,who seeked to the treatment in the dermatological departments from 112 hospitals in China.The survey consisted of several parts,including the general status of patients,the patients' cognition of occurrence,development and risk factors of acne,whether the first choice was seeking treatment at the hospital when the patients had acne and the condition of selection of skin care products.The factors were analyzed,which could impact the cognition of the patients' behavior of treatment,how did the patients' cognition to influence their medical behavior and skin care as well as the consistency of assessment of the severity of acne by doctors and patients themselves.Results The acne patients studied had the best knowledge of "acne is a skin disease","it not only occurs in the period of adolescence" and "the disease can be prevented and cured",which accordingly accounted for 80.65％,69.16％ and 65.49％ of the total patients respectively.However,the awareness of acne patients to heredity,high sugar and dairy products as risk factors for acne was insufficient,which accounted for 48.72％,42.40％ and 18.25％ of the total patients,respectively.Gender,age,educational level,occupation and health knowledge were the main factors affecting the cognitive level of patients;the survey also found that men,patient with educational level of junior high or even lower educational condition,occupation of labor workers or farmers and patients were lack of health education with poor knowledge of the genetics and dietary were risk factors for acne;patients with age over 36 years or with mild illness had poor knowledge of dietary risk factors for acne;the difference was statistically significant (P＜0.05).The analysis of the influence of cognitive status on medical treatment behavior and skin care showed that the better the cognition,the higher the probability of patients would choose medical treatment as the first choice as well as choosing functional skin care products;the difference was statistically significant (P＜0.05).The consistency of assessment of the severity of acne by doctors and patients was poor (Kappa value ＜0.4),and the assessment of severity of acne by patients was more serious than doctors' assessment.Conclusions Patient's cognitive status will affect their medical behavior and skin care,and there is also a phenomenon that patients have a more serious assessment of their acne condition.It is suggested that health education for acne patients should be strengthened in clinical medicine so as to improve their knowledge of acne as well as preventing from acne effectively.