Objective To study the effect of dendritic cells(DC) stimulated with C5b-9 on allogenic lymphocytes. Methods DC was stimulated with C5b-9.CD4+ T and CD8+ T cells were isolated by magnetic-activated cell sorting (MACS),and were cocultivated with DC.The maturation markers and cytokine secretion of T cells were analyzed by flow cytometry and ELISA.Results The high pure CD4+ T and CD8+ T cells were separated successfully.After cocultivated with DC,the muturation markers CD25 and CD69 of CD4+ T were up-regulated by 26.31% and 52.73%,and IL-2 and IFN-? of CD4+ T cells were promoted from 126.3 ng?L-1 and 156.7 ng?L-1 to 409.2 ng?L-1 and 471.5 ng?L-1;the levels of IL-10 before and after coculture were 104.3 ng?L-1 and 107.1 ng?L-1.However these markers and cytokine secretion of CD8+ T were not influenced.Conclusion C5b-9 can adjust the function of DC and regulate the immuological response of T cells.

Objective To investigate the potential effects and mechanism on peroxisome proliferator-activated receptor γ-toll-like receptor 4-tumor necrosis factor-α (PPARγ-TLR4-TNF-α) targeted pathway on hyperglycemia induced myocardium inflammation and oxidative stress. Methods Thirty-two Japanese healthy adult rabbits were randomly divided into four groups with 8 rabbits in each group: normal control group (NC group), diabetes mellitus group (DM group), diabetes mellitus + pioglitazone 4 mg·kg-1·d-1 and 8 mg·kg-1·d-1 groups (DM+PGZ 4 mg and 8 mg groups). DM model was reproduced by alloxan of 150 mg/kg through auricular vein injection. On the same day of successful DM model reproduction, the diabetic rabbits were fed with corresponding dose of pioglitazone in DM+PGZ 4 mg and 8 mg groups, but the rabbits in NC group were not challenged. After 8 weeks of feeding, venous blood of left jugular vein bifurcation and myocardium tissue were harvested respectively for the determination of inflammation and oxidative stress parameters. TNF-α, interleukin-1 (IL-1), adiponectin (ADP), nitric oxide (NO) and total nitric oxide synthase (NOS) levels were determined by enzyme linked immunosorbent assay (ELISA), myeloperoxidase (MPO) activity was determined by colorimetric method, superoxide dismutase (SOD) activity was determined by hydroxylamine method, malondialdehyde (MDA) was determined by thiobarbituric acid colorimetric method, and catalase (CAT) activity was determined by UV spectrophotometry method. In addition, the mRNA expressions of TNF-α and TLR4 were determined by real-time quantitate reverse transcription-polymerase chain reaction (RT-qPCR). Results ① IL-1 and TNF-α in serum and myocardium of model rabbits were significantly increased, ADP was significantly decreased, and the mRNA expressions of TNF-α and TLR4 in myocardium were significantly increased, indicating a significant inflammatory reaction. The inflammatory reaction in pioglitazone intervention groups was significantly reduced, TNF-αand IL-1 levels in serum and myocardium of DM+PGZ 4 mg and 8 mg groups were significantly decreased as compared with those of DM group [serum: TNF-α(ng/L) was 268.33±46.57, 261.34±33.73 vs. 331.40±69.05, myocardium: TNF-α (ng/L) was 144.72±26.90, 139.59±14.59 vs. 177.48±27.40; serum: IL-1 (ng/L) was 24.40±2.56, 23.35±3.13 vs. 30.08±5.44, myocardium: IL-1 (ng/L) was 21.26±2.85, 20.54±2.75 vs. 24.78±3.60, all P < 0.05], and ADP levels were significantly increased [serum (μg/L): 19.64±8.85, 20.54±7.47 vs. 15.45±3.06, myocardium (μg/L): 10.31±2.22, 11.49±3.42 vs. 7.76±1.77, all P < 0.05], and the mRNA expressions of TNF-α and TLR4 in myocardium were significantly decreased (TNF-αmRNA: 0.15±0.05, 0.14±0.06 vs. 0.25±0.09; TLR4 mRNA: 0.57±0.17, 0.40±0.18 vs. 0.75±0.35, all P < 0.05). ②Oxidative stress in serum and myocardium of model rabbits was significantly increased, SOD, NO, and total NOS levels were significantly decreased while the serum CAT and MDA levels were significantly increased without effect on MPO. Compared with the DM group, SOD and NO levels in serum and myocardium were significantly increased in DM+PGZ 4 mg and 8 mg groups [serum: SOD (U/L) was 571.39±40.85, 609.28±54.47 vs. 535.10±37.08, myocardium:SOD (U/mg) was 55.74±8.12, 53.60±9.87 vs. 42.26±12.34; serum: NO (μmol/L) was 2.95±0.51, 2.99±0.43 vs. 2.03±0.78, myocardium: NO (nmol/mg) was 1.95±0.37, 2.11±0.26 vs. 1.56±0.33, all P < 0.05], the serum MDA levels were significantly decreased (μmol/L: 20.11±2.34, 19.70±2.02 vs. 23.07±3.06, both P < 0.05), while no significant effect on CAT. There was no significant difference in parameter of inflammatory and oxidative stress between the two pioglitazone intervention groups. Conclusion 4 mg·kg-1·d-1 pioglitazone could activate PPARγ-TLR4-TNF-α targeted pathway, thus inhibit inflammatory and oxidative stress factors expression, and down-regulate hyperglycemia induced myocardium inflammatory and oxidative stress level, but the effect did not show a dose dependent manner.