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 investigate the effects of long-term low-dose radiation (LDR) of γ-rays on the proliferation and radiosensitivity of human lymphoblast cells HMy2.CIR (HMy) and to elucidate the underlying mechanism.Methods HMy cells were divided into control group and long-term LDR group.For the long-term LDR treatment,HMy cells were fractionally exposed to a low dose of γ-rays,which could enhance cell proliferation,3 times per week for 4 weeks.After the long-term LDR exposure,part of the control and long-term LDR exposed cells were further irradiated with a challenging dose (2 Gy) of γ-rays.Then cell proliferation and radiosensitivity were assayed by CCK-8 kit,cell apoptosis,and γ-H2AX formation was measured by flow cytometry.Gene expressions of cyclinD1,PCNA,bcl-2 and bax were detected by RT-PCR.Results The long-term LDR significantly increased cell proliferation (t =9.607,P ＜ 0.01) accompanied with up-regulation of cell cycle regulation gene cyclinD1 (t =6.869,P ＜ 0.01),proliferation regulation gene PCNA (proliferating cell nuclear antigen) (t =9.229,P ＜ 0.01) and bcl-2 gene (t =2.662,P ＜ 0.05),but decreased the expression of pro-apoptotic gene bax (t =19.908,P ＜0.01) in HMy cells.Compared to untreated cells,the long-term LDR decreased cell radiosensitivity (t =8.896,P ＜ 0.01),including apoptosis induction (t =4.762,P ＜ 0.01) and γ-H2AX formation (t =10.264,P＜0.01).Conclusions The long-term LDR promoted cell proliferation by up-regulating cell cycle related genes,while it reduced the radiosensitivity of HMy cells with acquisition of apoptotic resistance.

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 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 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 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.

The changes in excitability and autorhthmicity of bladder detrusor in experimental non-insulin dependent diabetes mellitus (NIDDM) rats were observed. Sixty-nine NIDDM rats as NIDDM group and 69 normal rats as control group were enrolled into this experimental study. At 6th, 10th, 14th, 18th, 22nd and 26th week after the rats were injected last time, the changes in the excitability and autorhthmicity of detrusor strips in vitro were observed. The results showed that the threshold of the tension which made the detrusor strips contract was significantly higher in NIDDM group (0.716 +/- 0.325 g) than in control group (0.323 +/- 0.177 g) (F = 59.63, P < 0.001). At different stages, the threshold of the tension resulting the contract of the detrusor strips in NIDDM group was also higher than in control group. At 18th week after STZ injection, the frequency of spontaneous contract of the detrusor strips in NIDDM was significantly higher than in control group (P < 0.05), whereas at 22nd week, that in NIDDM group was significantly lower than in control group (P < 0.05). It was concluded that the decreased excitability of the bladder detrusor was the earliest and most obvious changes in bladder function in diabetes rats and the autorhthmicity had also changed at the early stage of diabetic bladder.
Diabetes Mellitus, Experimental/*physiopathology ; Diabetes Mellitus, Type 2/physiopathology ; Muscle Contraction/physiology ; Muscle Relaxation/physiology ; Rats, Wistar ; Urinary Bladder/*physiopathology ; Urinary Bladder Diseases/etiology ; Urinary Bladder Diseases/*physiopathology