Objective:To investigate the expression of inducible costimulatory ( ICOS) and inducible costimulatory ligand ( ICOSL) on peripheral blood mononuclear cells ( PBMCs ) and their clinical relationship with rheumatoid arthritis ( RA ) patients.Methods:Peripheral blood samples were collected from 85 RA patients and 50 HC in this study.Expression of ICOS and ICOSL on PBMC from the subjects were detected by flow cytometry and real-time polymerase chain reaction( RT-PCR).The alteration of ICOS and ICOSL were observed after hormone therapy in 15 patients with RA and the relationship between their expression level and patients′clinical manifestations were analysed.Results:The ICOS and ICOSL mRNA level of RA patients′PBMCs were significantly higher than that in HC.The expression level of ICOS on CD4+T cells was higher than than that in HC[(7.08±4.72)% vs (3.01+1.39)%,P<0.0001].The expression of ICOSL on monocytes[(5.77±3.45)%vs (3.64±1.43)%,P<0.05] and B cells [(5.78± 4.52)%vs (3.97±1.63)%,P<0.05] were significantly elevated in RA patients.In RA patients with active disease,however,ICOSL expression on monocytes and B cells were increased as compared with those in inactive RA patients [ ( 5.45 ±3.50 )% vs ( 4.04 ± 1.55)%,P=0.036],[(6.59 ±5.74)%vs (5.63±4.30)%,P=0.016].Furthermore,after receiving immunosuppressive therapy, the expressions of ICOS and ICOSL were notably reduced as compared with pre-therapy levels on PBMCs from patients [ ( 5.45 ±3.50)%vs (4.04±1.55)%,P=0.036],[(6.59 ±5.74)%vs (5.63±4.30)%,P=0.016].Conclusion:The high levels of ICOS and ICOSL expression were closely correlated with the degree of disease and therapeutic response,suggesting that ICOS/ICOSL pathway may play a critical role in pathogenesis of RA.

Objective To investigate the expression of inducible co-stimulator (ICOS) and inducible co-stimulator ligand (ICOSL) on PBMCs,and the plasma concentrations of soluble forms of ICOSL and their clinical relationship with systemic lupus erythematosus (SLE) patients.Methods Peripheral blood samples were collected from 45 SLE patients and 39 healthy subjects (HC).The expressions of ICOS and ICOSL on peripheral blood mononuclear cells (PBMCs) were detected by flow cytometry.The concentrations of soluble ICOSL were assessed by measured by enzyme linked immunosorbent assay (ELISA).And the relationship between their expression levels and patients' clinical manifestations were analyzed.Levene F test was used for statistical analysis,the comparison between groups was conducted using t test,and the correlation between two variables were tested by Pearson correlation analysis.Results The expression of ICOS on CD4+ and CD8+ T cells were significantly higher than that of the HC [(19.1±1.7)％ vs (14.0±1.5)％,t=2.156,P=0.035],[(10.0± 1.0)％ vs (6.4±1.0)％,t=2.587,P=0.012].The expression of ICOSL on CD14+ mononuclear cells in SLE patients was significantly higher than that in the HC group [(2.94±0.88)％ vs (0.89 ±0.21)％,t=2.152,P=0.04].Plasma ICOSL concentrations in patients with active SLE were significantly higher than those of patients with inactive SLE [(362±25) ng/ml vs (278±15) ng/ml,t=2.356,P=0.025].We also found a significant negative correlation between the soluble ICOSL expression and the surface ICOSL expression on both mononuclear cells and B cells (r=-0.4243,P=0.022;r=-0.4099,P=0.025).MMPI induced an evident reduction in sICOSL levels released from the cells,which was statistically significant in comparison with untreated cells (P＜0.05).Conclusion The up-regulated expressions of ICOS and ICOSL on peripheral lymphocytes and the high levels of plasma concentration of soluble ICOSL are closely correlated with the severity of the disease,suggesting that ICOS/ICOSL pathway may play a critical role in the pathogenesis of SLE.

ObjectiveTo explore the relationship between the negative co-inhibitor programmed death-1 ( PD-1 ) and the pathogenesis of myasthenia gravis ( MG), by detecting the expression of PD-1 and programmed death ligand-1 ( PD-L1 ) on peripheral blood mononuclear cells (PBMCs) and soluble PD-1 (sPD-1) in plasma from myasthenia gravis patients. MethodsPeripheral blood samples were collected from 45 MG patients and 33 healthy persons without prednisone or other immunodepressant treatment during the half year ahead of withdrawal.The expression of PD-1 and PD-L1 on PBMCs were detected using immuno-fluorescence labeling and flow cytometry, and the concentrations of sPD-1 in plasma were measured using an ELISA kit. Results(1) The proportion of CD4+ PD-1 + T cells, as well as CD14+ PD-L1 +monocytes of the MG group was higher than that of the control group. There were no significant differences in the proportion of CD4+ PD-1 + T cells or CD14+ PD-L1 + monocytes in the MG sub-groups between different genders or MG types. While the proportion of CD4+ PD-1 + T cells of the late-onset MG (age ≥40) group was higher than that of the early-onset MG group (age ＜40). And it was higher in the MG patients with thymoma or thymus hyperplasia than that from the MG patients with normal thymus. The proportion of CD14+ PD-L1 +monocytes from the MG patients with thymoma or thymus hyperplasia group decreased obviously compared with that of the patients with normal thymus group; but no difference could be found between the late-onset group and early-onset group. (2)The concentration of sPD-1 in the plasma from the group of MG patients was(6. 92 ±0. 72) ng/ml,which was higher than that of the healthy control group ( (3.28 ±0. 42) ng/ml),even more, it was significantly higher in the early-onset MG group than that of the late-onset MG group,there was a negative correlation( r =-0. 526, P =0. 000) between the age of onset and the concentration of sPD-1. ConclusionsThe increased expressions of PD-1 on CD4+ T cells and PD-L1 on CD14+ monocytes in MG patients suggested the involvement of the couple of molecules in the pathogenesis of MG.Higher concentration of soluble PD-1 in the plasma of patients with MG suggested that it might disturb the ligation of PD-1 and PD-L1 on T cells and antigen presenting cells, which might result in the abnormal transportation of the negative modulating signal, and accelerate the pathological progress of MG.

Ischemic stroke is one of the primary causes of death and disability worldwide. Neutrophils can release depolymerized chromatin and proteins to form neutrophil extracellular traps (NETs) and participate in intravascular thrombus formation. In recent years, NETs have received increasing attention in the study of acute ischemic stroke. The results indicate that NETs play an important role in the pathogenesis of acute ischemic stroke. This review presented recent advances in the study of NETs in acute ischemic stroke.

Objective:To investigate the expression of costimulatory molecule OX40 in peripheral CD4 +CD25 +CD127 low regulatory T (Treg) cells and its clinical significance in patients with acute cerebral infarction (ACI). Methods:Seventy-five patients with first-onset ACI, admitted to our hospital from April 2019 to December 2019, and 36 age- and gender-matched volunteers (control group) were selected in this study. OX40 expression on CD4 +CD25 +CD127 low Treg cells in peripheral blood samples in the two groups were analyzed by immunofluorescent labeling and flow cytometry. Correlations of OX40 +Treg cell percentage with National Institute of Health stroke scale (NIHSS) scores, ischemic penumbra volume, core infarct volume, and infarct volume in the patient group were analyzed. The changes of OX40 +Treg cell percentage in the patient group before and after endovascular treatment or intravenous thrombolysis were compared. Results:As compared with that in the control group, the Treg cell percentage in peripheral blood samples of the patient group was significantly decreased, while OX40 +Treg cell percentage was significantly increased ( P<0.05). OX40 +Treg cell percentage was positively correlated with NIHSS scores in ACI patients ( r s=0.271, P=0.018). Meanwhile, OX40 +Treg percentage was significantly correlated with ischemic penumbra volume, core infarct volume, and infarct volume in the patient group ( r s=0.435, P=0.000; r s=0.343, P=0.003; r s=0.245, P=0.034). OX40 +Treg cell percentage in ACI patients 7 d after endovascular treatment was significantly decreased as compared with that before treatment ( P<0.05); OX40 +Treg percentage in ACI patients 3 and 7 d after intravenous thrombolysis was significantly decreased as compared with that before treatment ( P<0.05). Conclusion:OX40 is abnormally expressed on peripheral Treg cells in ACI patients, and is closely correlated with neurological deficits, imaging features and reperfusion therapy.

Objective To explore the expressions of costimulatory molecules OX40 and OX40L in peripheral blood (PB) samples of multiple sclerosis (MS) patients,and reveal their clinical significance.Methods PB samples of 39 MS patients and 37 health control subjects (HC) were collected from June 2013 to October 2018.OX40 expressions on CD4+ T cells,OX40L expressions on CD14+ monocytes and CD19+ B cells were detected by immunofluorescent labeling and flow cytometer.The correlations between expanded disability status scale (EDSS) scores and OX40 and OX40L expressions were analyzed,and the influences of different clinical typing and gender in OX40 and OX40L expressions were analyzed.The changes of OX40 and OX40L expressions before and after methylprednisolone shock therapy in 8 patients with recurrent MS were observed.Results (1) As compared with those in the HC group,the expressions of CD4+OX40+ and CD 14+OX40L+ in the peripheral blood of patients in the MS group were significantly increased (P＜0.05).(2) The OX40 expression on the surface of CD4+ T cells was positively correlated with EDSS scores of MS patients (r=0.684,P=0.000).(3) As compared with that in the MS patients at remission phase,the OX40 expression in the CD4+ T cells of MS patients at recurrence phase was significantly increased (P＜0.05);there were no significant differences in the expressions of OX40 and OX40L between male and female MS patients (P＞0.05).(4) EDSS scores of 8 patients with relapsed MS after intravenous infusion of methylprednisolone were significantly lower than those before treatment (P＜0.05);the OX40 expression on the surface of CD4+ T cells after intravenous infusion of methylprednisolone was significantly decreased as compared with that before treatment (P＜0.05).Conclusion Abnormal OX40 expression is closely correlated to neurologic impairment and clinical characteristics,and distinctly responded to corticosteroid therapy in MS patients,which suggests that OX40 may be a promising biomarker and is involved in the pathogenesis of MS.