0 05) Conclusion Intramuscular electroporation of pcD 2/Ang 1 can ameliorate the vessel leakage caused by pcD 2/VEGF and thus may represent a feasible and safe therapeutic strategy for patients with tissue ischemia

Objective To investigate the mechanisms of estrogen in the pathogenesis of systemic lupus erythematosus(SLE),we compared the effects of 17β-estradiol(E2)on bone marrow(BM)-derived dendritic cells (BMDCs)in(NZBxNZW)F1(NZB/w F1)female mice befbre and after the disease onset.Methods Immature,BMDCs were differentiated from BM monocytes cultured with rmGM-CSF,rmlL-4,E2 and estrogen receptor (ER)modulator-tamoxifen(TAM)for 7 days.Then immature DCs were stimulated by LPS for 24h to get mature BMDCs.Flow cytometry was utilized to detect the production of costimulatory molecule CD40 and intracellular IL-6,IL-10,IL-12 and TNFα of DCs.Mixed lymphocytes reaction was used to measure the stim-ulatory activity of DCs on spleen T lymphocytes.Results E2 mainly increased the expression of CD40 on im-mature BMDCs but reduced its expression on mature BMDCs.E2 enhanced the stimulatory activity of immature BMDCs,but weakened the activity of mature BMDCs.E2 decreased the production of cytokines of BMDCs in young mice,but increased them in old mice.Conclusion E2 modulates the functions of BMDCs of lupus model mice by binding with ER.The modulation varies depending on disease progression and cell maturation status.

Objective To assess the effects of protease 3(PR3)and protease-activated receptor (PAR)-2-activator on the maturation and functions of peripheral blood dendritic cell(DC)-like monocytes.Methods Density gradient centrifugation was used to isolate peripheral blood mononuclear cells(PBMC)from Wegener's granulomatosis(WG)patients and healthy controls(HC).PBMC were stimulated by LPS,human PR3,trypsin,PAR-2-agonist peptide (PAR-2-AP),LPS+PR3 or LPS+trypsin for 24 h.Flow cytometry was used to analyze the expression of PAR-2,CD80,CD83,HLA-DR on stimulated DC-like monocytes-CD14+CD16high monocytes.ELISA kit was used to test the concentration of IL-6 in the culture supernants.Mann-Whitney non-parameteric test was used fur statistical analysis.Resuits No effect of PR3,trypsin and PAR-2-AP on the expression of PAR-2,CD80,CD83,HLA-DR of DC-like monoeytes was found.LPS could significantly induce PAR-2 expression in HC[from(5.8±1.5)%to(24.5±4.5)%,P=0.002]and the expression of CD80,CD83,HLA-DR in HC and WG;PR3,trypsin,PAR-2-AP and LPS could all stimulate the secretion of IL-6.Conclusion PR3 and PAR-2 pathway-activators can not promote PAR-2expression and maturation of DC-tike monocytes,but they can induce the secretion of IL-6.

Objective To investigate the effect of estrogen on dendritic cells (DCs) and its role in disease progression by comparing the effects of 17β-estradiol (E2) on spleen DCs in systemic lupus erythe matosus (SLE) murine modeI-(NZBxNZW) F1 (NZB/w F1 ) female mice before and after the disease onset.Methods Anti-CD11c antibody labeled magnetic bead was used to purify spleen DCs.Flow cytometry was utilized to detect the co-stimulatory molecules and intracellular cytokines of DCs.Mixed lymphocytes reaction (MLR) was used to measure the stimulatory activity of DCs to T iymphocytes.And semi-quantitative RT-PCR was employed to check the expression of estrogen receptor (ER).Results E2 could modulate the expression of surface molecule CD40,the production of cytokines IL-6,IL-10,IL-12 and TNFa,and the stimulatory a bility of spleen DCs in SLE model mice.Tamoxifen (TAM) could antagonize E2 effects and E2 could affect the estrogen receptor (ERα) level of DCs.These changes of DCs varied with age.Conclusion E2 may be in volved in the pathogenesis of SLE by modulating DCs by binding ERa.The effects of E2 vary in different stages of SLE progression.

Objective The aim of this study was to assess the status of protease-activated receptor-2(PAR-2) expression on the peripheral blood immune cells including dendritic cells(DC) of Wegener's granulo matosis(WG) patients.Methods Flow cytometry was used to analyze the expression of PAR-2 protein on peripheral blood immune cells,DC and DC-like monocytes of healthy controls (HC),inactive,active and different medicine-treated WG patients.Two-tail Mann Whitney non-parametric test was used for statistical analysis.Results There was no difference of PAR-2 expression on PMN,monocytes,lymphocytes and DC between WG patients and HC.However,PAR-2 expression on CD14+CD16+ (t=3.823,P=0.004) and CD64+CD16+ (t=2.652,P=0.024) DC-like monocytes was much higher in WG patients compared with HC.In active WG,expression of PAR-2 was up-regulated on the cell surface of PMN (t=2.690,P=0.013 ),monocytes (t=1.688,P=0.047),lymphocytes(t=1.742,P=0.038),BDCA3+DC(t=2.582,P=0.016),CD11c+DC(t=1.828,P=0.044),CD14+CD16+ (t=3.419,P=0.002)and CD64+CD16+ (t=3.494,P=0.005) DC-like monocytes when compared with inactive WG.PAR-2 expression on these cells was similar between cyclophosphamide (Cyc) and Methotrexate (MTX) treated WG patients.Conclusion PAR-2 expression on immune cells,DC and DC-like monocytes correlates with WG occurrence and development.There is no difference in the effects on PAR-2 expression between various medications.

Objective To evaluate the magneto-heating effect and cytotoxicity of the carbonyl iron powder as a feasibility for arterial embolization hyperthermia. Methods Different doses of carbonyl iron powder suspensions were prepared in vitro(10 mg/ml,100%;10 mg/ml,50%;or 64 g/L phenol solutions),and heated for 20 minutes in an alternating magnetic field(49.9,79.9,and 110.2 Oe).The influences of the doses of suspensions and currency of magnetic field on the heating effect were observed.Meanwhile,mouse fibroblast L-929 cells were cultured with the suspensions for 2,4,or 7 days.The morphology and relative growth rate(RGR) of the cells were determined by microscopy and MTT assay.The cytotoxicity of the suspensions was then classified. Results The heating ability of the carbonyl iron powder increased with the suspension concentration and the strength of the magnetic field.A optimal therapeutic temperature was achieved at 110.2 Oe with 60 mg/ml carbonyl iron powder suspension.The L-929 cells showed normal morphology after been treated by the carbonyl iron powder(10 mg/ml 100% solution and 50% solution) for 2,4 and 7 days with the 0-1 degree cytotoxicity.Conclusion The carbonyl iron powder has good heating effect under the alternating magnetic field,and is compatible with the tested cells.

Objective To study the clinical application of the complete audiologic evaluation battery consisting of ABR,ASSR and behavioral audiometry in sound field for the identification of sensorineural losses in children.Methods In the study 48 children were divided as group PTA of the children assessed by pure tone audiometry and ABR and ASSR test,and group BA of the children unable to perform on behavioral evaluations.Results The analysis of the audiometric results of group PTA showed significant correlation between the ASSR thresholds and pure tone thresholds(P＜0.01) at various frequencies.The correlation coefficients were 0.75,0.76,0.76,and 0.83 at different frequencies.The ASSR-PTA linear regression was utilized.The comparison of the ABR and ASSR results of group BA indicated that 23 ears responded in ASSR but without responses in ABR.Those without ASSR responses also had no ABR There were 116 tests in total four frequencies for the 29 ears.83 responses were obtained from ASSR,89 responses from behavior audiometry and 96 from the use of the both tests.Conclusion The comprehensive audiologic evaluation battery could be used to test younger children with severe heating loss children as compared to the pure tone audiometry alone.More detailed data can be obtained thus from children to assist in rehabilitation in a more effective mariner.

Objective Among the factors affecting embryo implantation, oxidative stress is one that receives much medical attention. The purpose of the study was to explore the effect of H2 O2 on the endometrial stromal cells ( ESCs) in the decidual mouse model of oxidative stress of ESCs. Methods Primary ESCs cultured by enzymatic digestion with a mesh filter were divided into a control, an experimental, and a model group. Decidualization was induced in the mice of the experimental and model groups by inter-vention with 10 nM E2 and 1μmol/L P4 for 72 h in vitro. Then, the animals of the model group were treated with different concentra-tions of H2 O2 for 4 hours. The primary ESCs were identified by immunohistochemistry, the expression of dPRP mRNA determined byRT-PCR, the proliferation of the decidual ESCs treated with H2 O2 analyzed by CCK-8, and the level of ROS detected by flow cytometry. Results Primary mouse ESCs were successfully isolated, cultured, and identified, which were shaped like spindles and polygons, radial-ly aligned under the microscope. Immunofluorescence analysis showed positive expression of vimentin and negative expression of cytokeratin. The purity of the primary mouse ESCs was ( 96. 3 ± 0. 49 )%. Theexpression of dPRP mRNA was significantly higher in the ESCs treated with E2 and P4 than in the control (0.0002±0.0000 vs 1.0010±0.0011, P<0.01). H2O2 at ≥150μmol/L suppressed the proliferation of the decidual ESCs by 6.9% (P<0.05), and in a concen-tration-dependent manner, reaching the maximum inhibition rate of 70.6% at the concentration of 300μmol/L. The level of intracellular ROS was markedly increased in the ESCs treated with H2O2 at 50μmol/L (27.77±4.20) and 100μmol/L (43.57±6.58), with statisti-cally significant difference from that in the control group (17.47±0.61) (P=0.001). Conclusion H2O2 at 100μmol/L can signifi-cantly elevate the intracellar ROS level without affecting the proliferation of decidual primary ESCs, and therefore can be used for estab-lishing the model of oxidative stress of ESCs in decidual mice.

Objective To study and compare the level of adiponectin in different functional status of thyroid,to assess the association of thyroid hormones and insulin resistance with adiponectin.Methods ELISA was used to examine the fasting plasma adiponectin in 56 hyperthyroidism,30 hypothyroidism and 44 normal control subjects.FT_3,FT_4,TSH, FPG,Fins and blood fat were measured.BMI and HOMA-IR were calculated.Results There were higher level of adiponectin in hyperthyroidism than hypothyroidism and normal control subjects.Fasting plasma adiponectin was posi- tively correlated with FT_4 (r=0.36,P