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Objective To investigate how gene silence of CD40 by small interfering RNA(siRNA)influences the balance between Th1 / Th2 and Th17 / Treg in rats with experimental autoimmune myocarditis(EAM). Methods Lewis rats were divided into a normal - control group,EAM group,CD40 siRNA - therapy group and siRNA - control group by using random number table. EAM,CD40 siRNA - therapy and siRNA - control groups were immunized on day 0 and day 7 with purified porcine cardiac myosin to establish EAM,and CD40 siRNA was administered intravenously on day 7. The basic data of each group were observed,and the rats were executed on day 21 to study the pathology of myo-cardial tissues and the myocardial histopathology scores were calculated,and than the changes in electrocardiograms (ECG)and echocardiogram were recorded. Enzyme - linked immunosorbent assay(ELISA)was used to determine the levels of interferon(IFN) - γ,interleukin(IL) - 10,IL - 17 and transforming growth factor(TGF)- β in peripheral blood. Results (1)The overall incidence of EAM,CD40 siRNA - therapy and siRNA - control group was 100% ,and no rats died from day 0 to day 21.(2)There were only 2 premature rats in EAM and siRNA - control groups,and the ECGs of normal - control group and CD40 siRNA - therapy group were normal.(3)The echocardiogram of EAM,CD40 siRNA - therapy and siRNA - control group,displayed the left ventricular dilatation,hypertrophy of the left ventricle, the weakening of the left ventricular wall motion,and heart failure,and pericardial effusion was discovered. The left ven-tricular fractional shortening(LVFS)and the left ventricular ejection fraction(LVEF)of rats treated by CD40 siRNA were higher than those of rats in the EAM group[(46. 70 ± 8. 25)% vs(34. 28 ± 11. 81)% ,(80. 92 ± 11. 76)% vs (64. 03 ± 12. 60)% ,F = 0. 652,0. 234,all P ﹤ 0. 05].(4)Pathologic examination of the EAM group,CD40 siRNA therapy group and siRNA control groups,showed myocardial fiber fracture,myocardial tissue necrosis and inflammatory cell infiltration. But myocardial tissue pathological scores of the cardiac tissue of CD40 siRNA therapy group was lower than those of the EAM group(2. 10 ± 1. 07 vs 3. 40 ± 0. 72,F = 1. 290,P ﹤ 0. 05).(5)ELISA method examination re-vealed that the levels of IFN - γ,IL - 4,IL - 17 and TGF - β in EAM group were increased compared with those of nor-mal control group[(1 245. 55 ± 244. 56)ng/ L vs(501. 53 ± 18. 93)ng/ L,(78. 03 ± 10. 47)ng/ L vs(30. 77 ± 1. 74)ng/ L,(80. 82 ± 13. 33)ng/ L vs(27. 98 ± 3. 01)ng/ L,(156. 27 ± 12. 11)ng/ L vs(4. 33 ± 0. 79)ng/ L,t =7. 408,10. 764,9. 250,30. 608,all P ﹤ 0. 05]. However,the levels of IFN - γ and IL - 17 in CD40 siRNA therapy group were lower than those in the EAM group[(940. 62 ± 128. 40)ng/ L vs(1 245. 55 ± 244. 56)ng/ L,(60. 42 ± 12. 40) ng/ L vs(80. 82 ± 13. 33)ng/ L,t = 2. 704,2. 745,all P ﹤ 0. 05],while the levels of IL - 4 and TGF - β were higher [(97. 91 ± 13. 62)ng/ L vs(78. 03 ± 10. 47)ng/ L,(178. 84 ± 12. 10)ng/ L vs(156. 27 ± 12. 11)ng/ L,t = 2. 835, 3. 229,all P ﹤ 0. 05]. Conclusions The administration of CD40 siRNA markedly reduces myocardial inflammation of EAM rats and improves their cardiac function,which can be explained by Th1 - type and Th17 - type cytokines inhibi-ted,Th2 - type and Treg - type cytokines increased,and so then the imbalance of Th1 / Th2 and Th17 / Treg corrected.
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Objective To explore the alternations of CD4 + T cell subsets and their cytokines and transcription factors in children with Henoch - Schonlein purpura(HSP). Methods Forty - one children were enrolled in this stud-y,including 41 in acute stage of HSP,35 cases of HSP in clinical remission stage and 30 healthy children as healthy control group. The percentages of helper T lymphocytes(Th)1,Th2,regulatory T cells(Treg)and Th17 subsets were determined by flow cytometry(FCM). The concentrations of plasma interferon - γ( INF - γ),interleukin( IL) - 4, transforming growth factor - β1(TGF - β1 )and IL - 17 were examined by ways of enzyme - linked immunosorbent assay(ELISA). The expressions of T - bet,GATA3,Foxp3 and ROR - γt mRNA in peripheral blood mononuclear cells were examined by means of reverse transcription - polymerase chain reaction(RT - PCR). Results (1)During the a-cute and recovery stage of HSP,compared with the control group,the percentages of Th2 and Th17 cell subsets,the le-vels of plasma IL - 4 and IL - 17,and the expressions of GATA3 and ROR - γt mRNA were significantly higher(all P ﹤ 0. 05). The percentages of Th1 and Treg cell subsets,the levels of plasma INF - γ and TGF - β1 and the expres-sions of T - bet and Foxp3 mRNA were lower(all P ﹤ 0. 05). The imbalances of Th1 / Th2 and Treg/ Th17 appeared during the acute stage of HSP.(2)During the recovery stage of HSP,the percentages of Th1,Th2,Th17 and Treg,and the ratio of Treg / Th17 were of no difference compared with those in acute stage( all P ﹥ 0. 05). The ratio of Th1 / Th2,and the expressions of T - bet and Foxp3 mRNA were increased(all P ﹤ 0. 05),while the levels of plasma IL - 4 and IL - 17,the expressions of GATA3 and ROR - γt mRNA were decreased(all P ﹤ 0. 05)compared with those in acute stage. In the recovery stage of HSP,the imbalances of Th1 / Th2 and Treg/ Th17 were still obvious .(3)There was a positive correlation in every 2 cytokines of Th1,INF - γ and T - bet. And the same correlation existed in Th2, IL - 4 and GATA3,in Treg,TGF - β1 and Foxp3,and in Th17,IL - 17 and ROR - γt(P ﹤ 0. 05). Conclusions The imbalance of Th1 / Th2 and Treg/ Th17 is critical in pathological mechanism of HSP. The disturbance of immune tole-rance induced by Treg cells is important in HSP.
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Objective To investigate the effect of continuous blood purification (CBP) on pro-or anti-inflammatory immune function and prognosis in patients with sepsis. Methods One hundred and two patients with sepsis were randomized into the CBP group (60 cases) and the control group (42 cases). The patients in the control group were treated with conventional therapy, and the patients in the CBP group received at least 72 h CBP treatment additionally. The APACHEⅡ score, SOFA score, the 28 day survival rate and ICU length of stay were recorded and levels of spleen Th1, Th2 were assessed by FACS flow cytometry. Enzyme linked immunoadsordent assay (ELISA) was used to determine the levels of serum IL-1, IL-10 and TNF-α before and at 24, 48, 72 h after the treatment. Results The APACHEⅡscore and SOFA score decreased markedly in the CBP group after the treatment (Р 0.05). Compared with the control group, levels of IL-1, IL-10 and TNF-α were decreased markedly, and the ratio of Th1 / Th2 was increased significantly at 72 h after the treatment in the CBP group (Р < 0.05). Conclusion CBP can eliminate inflammatory mediators, and help to enhance the immune function, and restore the balance of Th1 / Th2 in patients with severe sepsis.