Objective To correlate the neutrophil extracellular trap (NET) mitochondrial DNA (mtDNA) and anti-mtDNA antibodies (Abs) with disease activity and clinical features in systemic lupus erythematosus (SLE) patients.Methods We enrolled 102 SLE patients,rheumatoid arthritis (RA) patients (n=30) and healthy donors as controls (n=40).NET were generated from phorbol 12-myristate 13-acetate (PMA)-stimulated peripheral neutrophils.mtDNA levels and the transcriptional levels of five interferon inducible genes(IFIGs)(OAS-1,Mx-1,Ly6e,IFIT1 and IFIT4) were measured by quantitative PCR.Interferon scores (IFN scores) were calculated.Anti-mtDNA Abs were detected by enzyme-linked immunosorbent assay.Spearman's correlation analysis,t test andx2 test were used for statistical analysis.Results mtDNA release by netting neutrophils was greatly enhanced in SLE patients (1 088 000 ±1 133 000) compared with healthy controls (465 900±447 200)(t=2.617,P＜0.01) and significantly correlated with IFN scores (r=0.460 6,P＜0.01).NETs mtDNA in moderate active group (728 300±1 003 000) and severe active group (1 093 000±946 500) were significantly higher than the mild active group (159 500±155 100) (t=2.240,P＜0.05,t=3.894,P＜0.01).Forty-one percent of SLE patients were positive for anti-mtDNA Abs(1.28±0.68),while none of the healthy donors (0.70±0.31) (P＜0.01) and RA controls (0.59±0.18)(P＜0.01) displayed a positive serology response to mtDNA.Addition-ally,the titers of anti-mtDNA Abs were also associated with IFN scores (r=0.292 8,P＜0.05).Anti-mtDNA Abs in moderate active group (1.3±-0.6) and severe active group(1.4±0.7)were significantly higher than the mild active group (0.7±0.4) (t=3.154,3.538,all P＜0.01).The levels of anti-mtDNA Abs significantly correlated with classic an-ti-dsDNA Abs titers measured by Farr assay (r=0.542 9,P＜0.001) and were associated with LN (x2=8.644,P＜0.01).Conclusion mtDNA in NET and anti-mtDNA Abs serve as new biomarkers for disease activity and renal involvement in SLE patients.

OBJECTIVE:To investigate the effects of triptolide(TP)on the proliferation of fibroblast-like synoviocytes(FLS) from patients with rheumatoid arthritis(RA)in vitro. METHODS:5 RA patients received knee arthroplasty or synovectomy to ob-tain synovial tissue. FLS was isolated,cultured and identified,and then incubated in the presence of TP [0 (blank control),50, 100 and 200 nmol/ml] for 24,48 and 72 h. The effects of TP on FLS was evaluated by MTT,and then proliferation inhibitory rate was calculated;flow cytometry was used to detect the apoptosis and cell cycle of FLS. RESULTS:The inhibitory rates of TP(50, 100 and 200 nmol/ml)on the proliferation of FLS were 17.46%-52.56%,which was positively correlated with drug concentration. Compared with blank control group,100 and 200 nmol/ml TP could increase the percentage of cells in G0/G1 phase and decrease the percentage of cells in S phase,with statistical significance(P<0.05);200 nmol/ml TP could induce cell apoptosis. CONCLU-SIONS:TP could inhibit the proliferation and also could induce the apoptosis of FLS in RA patients in vitro,which may be one of its mechanism for treating RA.

Objective To assess the clinical significance of glucose-6-phosphate isomerase in RA patients. Methods The level of serum GPI in 100 patients with RA, 98 patients with other rheumatic diseases and 108 normal controls were assessed by sandwich ELISA methods. The level of RF, CRP, anti-CCP antibodies were also assessed in RA patients. Results The level of GPI was higher in RA patients [(2.4?5.0) ?g/ml] than that of normal control group [(0.12?0.14) ?g/ml (P

Objective To explore the adverse effect of arsenic trioxide (As2O3) on liver,kidney and heart function during treating children patients with acute promyelocytic leukemia (APL) at therapeutic dose.Methods Sixty-five APL cases received As2O3 by intravenous drip and organic toxicity were selected as our subjects.The indices of liver,heart and kidney were measured.Results Of all subjects,19 cases(29.2％) occurred liver damage,including 15 cases(23.1％) mild and 4 cases(6.2％) moderate toxicity.The levels of alanine aminotransferase of patients before treatment was (19.9 ±9.5) U/L,and (24.3 ± 11.8) U/L,(25.0 ± 14.4) U/L at 1 st and 2nd weeks after treatment,higher than those before the treatment (P ＜ 0.05).However,level of alanine aminotransferase was back to normal at 3th weeks after treatment.Meanwhile the levels of aspartate aminotransferase at 1st,2nd and 3th weeks after treatment were (38.3 ± 16.5),(39.1 ± 15.5),(35.3 ± 20.6) U/L respectively,higher than that before treatment((28.5 ± 8.8) U/L,P ＜ 0.05 or 0.01),and it was back to normal at 4th weeks.(2) The levels of urinary cystatin C were (2.51 ± 1.45) mg/L,(3.05 ± 1.13) mg/L,(2.46 ± 1.21) mg/L at 2nd,3th,4th weeks after treatment,significantly higher than that before treatment ((1.98 ±0.68) mg/L,P ＜0.05 or 0.01).And the levels of urinary β2 microglobulin at 2nd,3th,4th weeks after treatment were significantly higher than that before treatment (P ＜0.05 or 0.01) and back to normal at 5 weeks after treatment.(3) Nine cases at remission stage showed the symptoms of palpitation,precordial discomfort and increased heart rate,and all those symptoms were mild.And the symptoms disappear at the 3th week after the treatment.Creatine kinase at the 2nd weeks after treatment was (90.2 ± 32.5) U/L,higher than that before treatment ((78.5 ± 22.3) U/L).The levels of creatine kinase isoenzyme at 2nd,3th weeks after treatment were (8.3 ± 4.8) U/L,(8.5 ± 5.6) U/L,higher than that before treatment ((6.3 ± 3.5) U/L).The serum creatine kinase mass at 4th weeks((3.9 ±2.0) g/L) was significantly higher than that before treatment ((2.8 ± 1.9) g/L),and then gradually be back to normal.Conclusion The routine dose As2O3 in treatment of APL children show less toxicity in liver,kidney,and heart Those adverse effects are transient,reversible and they occurred at 1-3 week after As2O3 treatment.Serum alanine aminotransferase,aspartate aminotransferase and urinary cystine protease inhibitors,β2 micro ring protein and serum creatine kinase MB mass might be served as sensitive indicators of organ damage.

Objective To establish the method of testing immunoglobulin G (IgG) with Fc sialylation, and to investigate the clinical significance of IgG with Fc sialylation in systemic lupus erythematosus (SLE), especially in those with neuropsychiatric manifestations (NPSLE). Methods Seventy-five SLE including thirty patients with neuropsychiatric manifestations (NPSLE) and forty-five non-NPSLE patients, 30 rheuma-toid arthritis (RA) patients, 32 juvenile idiopathic arthritis (JIA) patients and 41 healthy controls were recruited in this study. Standard method of testing lgG with Fc sialylation was established by a lectin based sandwiched enzyme linked immunosorbent assay (ELISA). The levels of IgG with Fc sailylation were assayed, and its clinical significance was evaluated. Results There were no difference in the levels of serum IgG with Fc sailylation in RA group (0.82±1.81) mg/ml, JIA group (0.69±1.30) mg/ml, healthy control group (0.64± 1.09) mg/ml, and the levels of IgG with Fc sailylation in SLE group (0.12±0.17) mg/ml (P<0.01), especially in NPSLE group [(0.03±0.03) mg/ml, P<0.01] was significantly lower than that of control groups. The perc-entage of IgG with Fc sialylation in control group (4.64±5.90)% were significantly higher than that in non-NPSLE (1.88±2.16)% (P<0.01) and in NPSLE (0.29±0.47)% (P<0.01). The percentage of IgG with Fc sial-ylation was negatively associated with SLEDAI score (r=-0.43, P<0.01). Conclusion Significantly low level of serum IgG with Fc sialylation was associated with disease activity in SLE patients, especially in NPSLE patients, lgG with Fc sialylation may be a new target for therapeutic strategy.

Objective To investigate the expression of Signal transducer and activator of transcription 2 (STAT2) gene in the peripheral blood mononuclear cells of patients with systemic lupus erythematosus, and to evaluate the possible connections between STAT2 gene expression levels and clinical features. Methods One hundred and forty-four SLE patients, 27 non-SLE patients with other rheumatisms and 58 normal controls were recruited for this research, and the subjects were surveyed for clinical data collection. SYBR Green Dye based real-time quantitative PCR method was used to compare the expression levels of STAT2 in patients with SLE and those in the controls. The correlation of the gene expression levels and disease activity and specificity was studied. Results STAT2 expression levels (5.2±1.7) in SLE patients were remarkably higher than those in non-SLE patients and normal controls (4.3±1.1, 4.5±1.2, P<0.01 in both). The expression levels of STAT1 were increased in active SLE patients(5.2±1.5), comparing with those observed in inactive SLE patients (4.8±2.9, P<0.01), and expression levels of STAT1 in SLE patients were negatively correlated with C3 levels in sera (r=-0.449, P<0.01) whereas were positively correlated with SLEDAI-2K score and 24 hour urine protein (r=0.317, 0.309, P<0.01 in both). Conclusion Over-expression of STAT2 gene in the peripheral blood cells is linked with the pathogenesis of systemic lupus erythematosus, and the elevated expression level of STAT2 is correlated with SEE disease activity.

Objective Renal vascular injury,especially thrombotic microangiopathy (TMA),is an important prognostic factors in pat.ients with lupus nephritis.TMA is most likely to be associated with proliferative lupus nephritis.This single-center retrospective study was conducted in order to explore the characteristics and prognosis of patients with TMA associated with proliferative lupus nephritis.Methods From January 2013 to June 2016,146 hospitalized patients with lupus nephritis underwent renal biopsy in the Department of Rheumatology,South Campus,Ren Ji Hospital,of which 108 were proliferative nephritis including 32 with TMA and 76 without TMA.All the clinical records were collected.All data were analyzed by Graphpad 5.0 or SPSS 22.0 statistical software analysis.Nonparametric test,t test and Fisher test were used for comparison between the two groups.Predictors were analyzed by multiple factors regression analysis,survival curve was analyzed by Kaplan-Meire method.Results Patients with TMA were found to have higher levels of creatinine (Cr) [93.5 (69.0,179.8) μmol/L vs 73.0 (56.3,116.3) μmol/L,U=833,P=0.010 1],B-type brain natriuretic peptide (BNP) [177(93.2,619) pg/ml vs 87.5(28.5,255) pg/ml,U=765,P=0.004 6],24-hour urinary protein [4.98 (1.99,7.62) g vs 2.83 (1.71,4.38) g,U=875,P=0.022] and highersystemic lupus erythematosus disease activity index (SLEDAI) [16(13.25,18) vs 12(10.25,14),U=559,P＜0.000 1],as well as lower complement [C3:(0.37±0.15) g/L vs (0.52±0.20) g/L,t=3.713,P=0.000 3;C4:0.056 (0.035,0.140) g/L vs 0.088(0.053,0.167) g/L,U=912,P=0.047 9],albumin (Alb) [(24±6) g/L vs (28±6) g/L,t=3.416,P=0.000 9] and hemoglobin (Hb) [(88±19) g/L vs (99±21) g/L,t=2.627,P=0.015 7].They were more prone to hypertension [(24/32,75％) vs (35/76,46％),x2=7.613,P=0.006 4],had less glomerular sclerosis [0(0,0.038)％ vs 7(0,17)％,U=848,P=0.007 7] and higher acute score [16(14.25,19.75) vs 13(10,15),U=612,P＜0.000 1];while these patients received higher doses of corticosteroid therapy,and more patients were treated with cyclophosphamide for induction therapy.Multivariate regression analysis showed that Cr (OR =1.008,P=0.033) and SLEDAI (OR =1.272,P=0.003) scores might be predictors of TMA in patients with proliferative lupus nephritis.During follow up,6 and 2 patientsin two groups progressed to end-stage renal disease (ESRD),respectively (P=0.010 4).Univariate analysisfound that patients progressed to ESRD were more likely to have TMA[(6/8,75％) vs (23/85,27％),x2=7.831,P=0.010 4],and had more crescents[54.5(12,83.5)％ vs 20(6,41)％,U=183,P=0.032].Higher activity indices (AI) [(20±6) vs (14±4),t=3.489,P=0.000 7],Cr [286(214.5,395) μmol/L vs 76(59,115.5) μmol/L,U=19,P＜0.000 1] and BNP [456(192.3,1 060) vs 110(45.38,275.5),U=116.5,P=0.002 4],as well as lower Hb [(71±12) vs (97±19),t=3.776,P=0.000 3] and platelets (PLT) [(130±71)×109/L vs (196±76)×109/L,t=2.399,P=0.018 5] were observed in these patients.Conclusion This study has shown that patients with proliferative lupus nephritis with renal TMA have a higher level of Cr and more active disease state,requiring more aggressive immunosuppressive therapy and more likely to progress to ESRD.So renal TMA may be one of the risk factors of renal survival in these patients.

Objective To investigate autoantibody against endothelin receptor type A (ENRA-Ab) in patients with systemic lupus erythematosus associated pulmonary arterial hypertension (SLE-PAH).The possibility of autoantibody-mediated pathogenesis in the development of SLE-PAH has also been explored.Methods ENRA-Ab in the serum of SLE-PAH and controls were detected by using a human ETRA epitope peptide-based ELISA.The clinical relevance of ENRA-Ab in SLE-PAH was analyzed.Proliferation of vascular smooth muscle cells (SMCs) and permeability of endothelial cells in vitro under the stimulation of polyclonal ENRA-Ab IgG were assessed.The expressions of PAH-related markers, i.e., 5-HTT, PDGFR-b, VEGF-A and PDGF-B were measured by qPCR.The effect of ENRA-Ab in vivo was also determined in a suboptimaldose monocrotaline-induced model with the assessment of right ventricle hypertrophy index and pathology parameters.Independent t-test, Tukey-Kramer test of variance analysis and Pearson' s correlation analysis were used for statistical analysis.Results ENRA-Abs was presented in a higher occurrence in SLE-PAH (35/85,41％) compared with controls (0/60;0, 13/80, 16％).There was a significant correlation between ENRA-Ab and echocardiograph estimated pulmonary arterial systolic pressure (r=0.392, P=0.002) in SLE-PAH.ENRA-Ab could promote SMCs proliferation, disrupt endothelial barrier and up-regulate PAH-related markers expression,which could be blocked in the presence of ETR antagonist.ENRA-Ab aggravated right ventricle hypertrophy and vascular remodeling in vivo.Conclusion ENRA-Ab is a new biomarker, in SLE-PAH, which may mediate PAH development in SLE.

Aim To investigate if LPS increases the sterol regulatory element binding proteins ( SREBPs ) cleavage-activating protein ( SCAP )-SREBP2 expres-sion by activation of mTOR signal pathway in THP-1 macrophages , upgrading LDLr level , causing foam-cell formation .Methods THP-1 macrophages were incu-bated in serum free medium in the absence of 5 mg?L-1 LDL alone , or 5 mg? L-1 LDL plus 200 μg? L-1 LPS, or 5 mg? L-1 LDL plus 200 μg? L-1 LPS plus 10 μg? L-1 rapamycin .Morphological examination of macrophages was performed with Oil Red O staining . Expression changes of LDLr , SREBP2, SCAP, S6K1 and mTOR mRNA were detected by real time quantita-tive polymerase chain reaction ( PCR ) .Western blot was used to analyze protein expression changes of LD-Lr, S6K1 and mTOR.Translocation of SCAP-SREBP2 complex from the endoplasmic reticulum ( ER ) to the Golgi was determined by confocal microscopy .Results LPS enhanced transformation of THP-1 macrophages into foam cells by increased uptake of lipid as evi-denced by Oil Red O assay .LPS increased mRNA lev-els of LDLr, SREBP2, SCAP, S6K1 and mTOR ( P <0.05) .Rapamycin reduced the mRNA levels of LDLr , SREBP2,SCAP,S6K1 and mTOR induced by LPS ( P<0.05 ) .Western blot demonstrated that LPS also caused over-expression of protein of LDLr , S6K1 and mTOR(P<0.05).Rapamycin reduced the expression of protein of LDLr, S6K1 and mTOR induced by LPS ( P <0.05 ) .Confocal microscopy demonstrated LPS caused an escape of SCAP-SREBP2 complex from the ER to the Golgi .Rapamycin inhibited the translocation of SCAP-SREBP2 complex from the ER to the Golgi . Conclusions Inflammatory stress increases SCAP/SREBP2 expression by activation of mTOR signal path-way, resulting in an escape of SCAP-SREBP2 complex from the ER to the Golgi , furthermore elevating LDLr expression and causing foam-cell formation .Rapamy-cin reverses the activation of mTOR signal pathway and decreases lipid deposition in THP-1 macrophages in-duced by LPS .

Objective To preliminarily investigate the effect and possible mechanisms of Senecio cannabifolius Less.Ⅱ(FHC-Ⅱ) on perfluoroisobutylene (PFIB) inhalation-induced acute lung injury. Methods Totally 156 rats were randomly assigned to three groups: the control group, the PFIB group and the FHC-Ⅱ prevention group, with 32, 62 and 62 rats in each group respectively. The FHC-Ⅱprevention group were given FHC-Ⅱthree times per day at the dosage of 340 mg/kg before PFIB exposure. 1 h after the last time of FHC-Ⅱ administration, the FHC-Ⅱ prevention group were exposured to gaseous PFIB (0.2 mg/L) for 10 minutes in a static whole-body exposure inhalation system. The survival rate of the rats were recorded at 1, 2, 4, 8, 16, 24, 48 and 72 h post PFIB exposure;the lung index and total protein content in bronchoalveolar lavage fluid (BALF) were measured at 1 h, 2 h, 4 h, 8 h, 16 h and 24 h; IL-1β and IL-8 in sera were assayed by enzyme-linked immunosorbent assay (ELISA) at 1, 2, 4, 8 and 16 h post PFIB exposure and the histopathological examination of the lung tissue was performed at 8 h post PFIB exposure. Results FHC-II significantly reduced the content of the total protein in BALF, lung index and the levels of IL-1β and IL-8 in aera as well, and dramatically alleviated the histopathological changes in the lung tissue. Conclusion FHC-Ⅱ demonstrates some preventive effect on PFIB inhalation-induced acute lung injury in rats.