Objective:To investigate the effect of hUCMSC-exos on the expression levels of HDAC in different isotypes of RA FLSs, and to elucidate the possible mechanism of hUCMSC-exos on the apoptosis of RA FLSs by regulating HDAC.Methods:hUCMSC and hUCMSC-Exos were isolated and identified. RT-qPCR was used to detect the changes in HDAC mRNA expression levels in FLSs after hUCMSC-Exos intervention, and the most affected HDAC types were identified. Western blot was used to detect the levels of FLS HDAC1 protein and the expression levels of NF-κB p65 and phospho-NF-κB p65 (Ser 536) in the blank control group, hUCMSC group, hUCMSC-Exos group, Trichostatin A (TSA) group and HDAC1 Inhibitor (Pyroxamide) group. To investigate the effects of hUCMSC-Exos on HDAC expression and NF-κB activity in FLSs. Flow cytometry was used to detect the effect of hUCMSC-Exos on the apoptosis of FLSs. ELISA was used to detect the effects of hUCMSC-Exos on the secretion of TNF-α, IL-6, IL-1β and IL-8 by FLSs. Flow cytometry and ELISA were used to detect the apoptosis level and pro-inflammatory cytokine secretion level of RA FLSs in the blank control group, NF-κB Inhibitor (pyrrolidine dithiocarbamate (PDTC) group, hUCMSC-Exos group and PDTC+hUCMSC-Exos co-intervention group. Whether inhibition of NF-κB affects the regulatory effect of hUCMSC-Exos on RA FLSs was further explored. All experimental data conforming to the normal distribution were compared by one-way ANOVA. LSD- t test was used for pin-group comparison, and independent sample t test was used for two-sample comparison. Results:Cultured primary hUCMSC were adherently grown spindle-shaped cells, and hUCMSC-Exos were saucer-shaped membranous vesicles, both of which met the identification criteria. hUCMSC-Exos reduced the expression level of HDCA1 mRNA [(0.932±0.091), t=2.19, P<0.001] and protein [(0.204±0.012), t=8.28, P<0.001] in RA FLSs, and the inhibitory effect was stronger than that of hUCMSC ( t=1.09, P=0.009) and HDAC1 ( t=11.29, P=0.013) Inhibitor. hUCMSC-Exos increased the apoptosis rate of RA FLSs [(48.68±0.84)%, t=12.33, P<0.001]. hUCMSC-Exos reduced the secretion levels of TNF-α [(29.6±1.0)pg/ml, t=10.78, P<0.001], IL-6 [(20.1±0.7)pg/ml, t=7.96, P<0.001], IL-1β [(9.28±0.23)pg/ml, t=6.14, P<0.001] and IL-8 [(108.0±3.8)pg/ml, t=1.21, P<0.001] in the supernatant of RA FLSs. hUCMSC-Exos reduced the expression level of p-NF-κB-p65/NF-κB-p65 in RA FLSs(0.351±0.024, t=17.67, P<0.001), and its inhibitory effect was stronger than that of hUCMSC (0.515±0.064, t=8.07, P=0.009) and HDAC1 inhibitor(0.411±0.033, t=2.44, P=0.04). After use of NF-κB inhibitors, hUCMSC-Exos weakened the promotion of apoptosis of RA FLSs [(29.0±0.5)%, t=10.63, P<0.001] and weakened the inhibitory effect of IL-8 secretion in the supernatant of RA FLSs [(125.5±3.2)pg/ml, t=2.63, P=0.002]. Conclusion:hUCMSC-Exos can mimic maternal cells to effectively inhibit the aberrant expression of HDAC1 in RA FLSs. hUCMSC-Exos may affect the apoptosis of RA FLSs and the secretion of pro-inflammatory cytokines by inhibiting the HDAC1/NF-κB pathway.

Objective:To identify the characteristics of nailfold capillaroscopy (NFC) of systemic sclerosis (SSc) and investigate whether more severe peripheral microangiopathy at NFC were related to the development of SSc.Methods:① The study included 115 patients (60 cases with SSc and 55 patients with other connective tissue diseases). All patients were treated with neither prednisone nor immunosuppressive drugs within 3 months before enrollment. We collected the following data: age, disease duration, disease onset, mRSS, high-resolution chest tomography (HRCT), echocardiography, pulmonary function, nailfold capillaroscopy and routine laboratory assessments. ② All the NFC definitions were used for semi-quantitatively scoring and Cutolo's qualitative assessment. ③ The relationship between NFC changes and joint, visceral involvement and autoantibodies in SSc patients was analyzed. ④ T test, Rank sum test and chi-square test were applied to analyze data. Results:① According to Cutoloqualitative assessment of NFC, patients of SSc with active/late pattern ( n=52) were very common than other CTD ( n=21) ( Z=-3.853, P<0.01). ② According to semiquantitative assessment, the scores of loss of capillaries [(1.67±0.60) vs (0.72±0.46), t=8.347, P<0.01)], irregular enlarged capillaries [(1.22±0.88) vs (0.74±0.50), t=3.178, P<0.01)], hemorrhage [(0.30±0.39) vs (0.10±0.21), t=3.090, P<0.01)], disorganization of the microvascular array [(0.38±0.38) vs (0.18±0.32), t=2.729, P<0.01)] were significantly higher than CTD. ③ The NFC of SSc patients was significantly different from CTD. The number of capillary loss ( Z=-4.194, P<0.01), input capillary dimensions ( t=3.704, P<0.01), output capillary dimensions ( t=3.913, P<0.01), wide diameter of capillary ( t=4.586, P<0.01), tortuous capillaries ( Z=-2.677, P<0.01), gaint capillary ( χ2=8.040, P=0.013), effusion ( Z=-2.278, P=0.023) were more increased than CTD. ④ The NFC pattern of SSc with lung involvement were mainly active and late (66%, 33/50), whereas early and active pattern (60%, 6/10) for those without respiratory system involvement ( Z=10.114, P=0.045) . The NFC pattern of SSc patients with joint involvement were mainly active and late (75%, 12/16), whereas early and active (66%, 29/44) for those without joint involvement ( Z=5.550, P=0.057) . Conclusion:The NFC of SSc patients is significantly different from CTD. NFC may be a suitable tool for disease evaluation.

Objective:To analyze the clinical features of patients with cardic involvement in eosinophilic granulomatosis with polyangiitis (EGPA), and to enhance the understanding.Methods:We retrospectively reviewed the clinical data of 16 patients with EGPA with cardiac involvement in Bethune hospital of Shan-xi from Jan 2012 to Jun 2019. T test, rank sum test and χ2 test were used for statistical analysis. Results:16 patients with cardiac involvement. There were 11 males and 9 female. The age of 16 patients with cardiac involvement ranged from 14 to 82 years old, and the average age of onset was (58±14) years. Compared with patients without cardiac invo-lvement was (41±15) years, the difference between the two groups was statistically significant ( t=3.230 , P<0.01). The analysis suggested that age was related to whether or not cardiac involvement. Cardiac related clinical symptoms occurred in 4 patients (25%). One patient presented with cardiac involvement as the initial symptom. The other 12 patients presented abnormal electrocardiogram (ECG), cardiac ultrasound or cardiac magnetic resonance imaging (MRI), including 10 patients (62%) with abnormal ECG, 13 patients (81%) with abnormal cardiac ultrasound examination, and1patient with cardiac MRI suggesting endocarditis. Among 16 patients with EGPA cardiac involvement, 14 presented with pulmonary involvement, 10 cases with nasal involvement, 9 cases with perip-heral neurological involvement, 9 cases with skin involvement, 6 cases with gastrointestinal involvement, 2 cases with kidney damage. Eosinophils (EO) were increased in all 16 patients, with a median value of 2.46 (1.49, 3.94) ×10 9/L, and EO was associated with cardiac involvement. Analysis of perinuclear anti-neutrophil cytoplasmic antibo-dies (pANCA) positive rate showed that only 2 of the 16 patients were positive. There was statistically significant difference ( P=0.017) compared with the group without cardiac involvement (8 patients were positive). All 16 patients were treated with glucocorticoid, 12 patients received immunosuppressive therapy, and 10 patients were treated with cyclophosphamide. During the ollow-up, 1 case died of heart failure, 1 case died of cerebral hemorrhage, 3 cases were lost to follow-up, and the other 11 cases were all stable after discharge. Conclusion:EGPA patients have a high incidence of cardiac involvement, and all cardiac stru-ctures can be involved, and most cardic involvement happens in ANCA negative patients. Cardiac involvement is one of the factors with poor prognosis.

Objective To analyze the clinical features and treatment of connective tissue disease (CTD) complicated with acquired hemophilia A (AHA).Methods A retrospective analysis of 8 cases of CTD [5 cases of systemic lupus erythematosus (SLE),2 cases of Sj(o)gren's syndrome (SS),1 case of rheumatoid arthritis (RA)] related to clinical manifestations,diagnostic methods,treatment options and outcomes.Results At the onset of AHA,active disease was shown in 7 patients with CTD,and 5 cases had bleeding symptoms in different parts.There were 3 cases of anti-phospholipid syndrome in 5 cases of SLE,2 of which had thrombosis.In 8 patients,the activated partial thromboplastin time (APTF) was prolonged by 1.7 to 3.times,FⅧ∶ C was 9.2％ to 21％ (50％ to 150％),and the factor Ⅷ inhibitor titer was increased by 7.6 to 56 BU/m1 (Bethesda method).Seven patients were treated with sufficient hormones,immunosuppressive agents,human immunoglobulin (IVIG),and blood products.Five patients had clinically improved bleeding tendency and APIT,and one patient was ineffective.Conclusion CTD is easy to combine with AHA.Glucocorticoid combined with immunosuppressive agent can effectively treat CTD-related AHA.For refractory patients,rituximab can be an alternative.

Objective To observe the effects of intra-articular ozone injection on the secretion of tumor necrosis factor (TNF)-α,Interleukin (IL)-6,IL-17A,and vascular endothelial growth factor (VEGF) in the serum of rats with collagen-induced arthritis (CIA) and explore the therapeutic mechanism of ozone in RA treatment.Methods Thrity-two Wistar rats were randomized into 4 groups,including the ozone groups that receivedintra-articularinjection of 40 μg/ml ozone (O3 group),a blank control group (normal group),a methotrexate (MTX) group (MTX group) anda collagen-induced arthritismodel (CIA group).All the rats,except for those in the blank control group,were subjected to hypodermic injection of bovine collagen Ⅱ and complete Freund's adjuvant to induce CIA.Ozone treatment was administered once weekly for 3 weeks starting at 14 days after the model were established.MTX group were treated with methotrexate 0.9 mg/kg,once a week,a total of three weeks.The swelling degree of the foot were observed,and the serum contents of TNF-oα,IL-6,IL-17A and VEGF were detected.One-way analysis of variance or Kruskal-Wallis test was used to evaluate the experimental data.Results At the end of treatment,the degree of foot swelling was reduced significantly in rats with O3 group compared with that in the CIA group [(4.21±0.14) ml vs (9.12±0.17) ml,t=8.43,P=0.023].The serum concentration of TNF-α,IL-6 and VEGF showed significant difference between the CIA group and the O3 group[91.55(86.55,98.53) pg/ml vs 14.45 (12.55,16.15) pg/ml,x2=6.216,P=0.002;145.08(37.44± 362.82) pg/ml vs 5.84(5.47,15.93) pg/ml,x2=13.136,P=0.004;51.56(46.09,74.10) pg/ml vs.36.22(32.18,41.69) pg/ml,x2=3.732,P=0.002].There was no statistically significant difference between the O3 group and MTX group [14.45(12.55,16.15) pg/ml vs [12.45(11.80,15.60) pg/ml,x2=0.243,P＞0.05;5.84(5.47,15.93) pg/ml vs 7.86(5.25,15.23) pg/ml,x2=0.058,P＞0.05;36.22(32.18±41.69) pg/ml vs 40.17(35.47,50.73) pg/ml,x2=0.516,P＞0.05].The serum concentration of IL-17A showed no significant difference between the normal group,the CIAgroup,the MTX group and the O3 group (F=1.827,P=0.165).Conclusion Intra-articular injecfion of 40 μg/ml ozone can attenuate synovitis in rats with CIA,the mechanism may relate to the inhibition of TNF-oα,IL-6 and VEGF in the serum.

Objective To evaluate the value of 2012 classification criteria for early rheumatoid arthritis (ERA),2010 American College of Rheumatology/European League Against Rheumatism (ACR/EULAR) classification criteria,and 1987 ACR classification criteria in the diagnosis of early rheumatoid arthritis (RA).Methods Patients who had at least one swollen and tender joint with disease duration no more than 2 years,and age more than 16 years were enrolled.The patients were diagnosed as RA or other non-RA by 2 experienced rheumatologists.The clinical and laboratory parameters were recorded.The sensitivity and specificity of three RA classification criteria were compared by McNemar test,The areas under the receiver operating characteristic curve (ROC) curve (AUC) of each RA classification criteria were analyzed using MedCalc software.Results Atotal of 310 patients were enrolled in this study,including 182ERA and 128 non-RA.The sensitivity(88.5％) of ERA criteria was much higher than that of the 1987 ACR criteria (45.6％,x2=75.013,P＜0.05),and not significantly different with the 2010 ACR/EULAR criteria (91.8％,X2=1.042,P＞0.05).The specificity of ERA criteria (91.4％) of 2010 ACR/EULAR criteria (87.5％,x2=1.8,P＞0.05) was similar to that of the 1987 ACR criteria (96.1％,x2=3.1,P＞0.05).The AUC of ERA criteria was 0.962 [95％CI(0.934,0.980)],which was slightly better than that of the 2010 ACR/EULAR criteria 0.959 [95％CI(0.931,0.978)],Z=0.380,P=0.7038,and much higher than that of the 1987 ACR criteria 0.885 [95％CI (0.845,0.919)],Z=4.517,P＜0.01.Conclusion Overall evaluation,the diagnostic value of ERA criteria is better than 1987 ACR and 2010 ACR/EULAR criteria in early rheumatoid arthritis.Compared to 2010 ACR/EULAR classification criteria,ERA criteria is more simple and practical.

Objective Endothelial microparticles (EMPs) are direct indicator of endothelial cell activation or apoptosis,and may also reflect endothelial inflammation,increased coagulation,and vascular tone.The aim of this study is to investigate whether EMPs would be able to evaluate systemic involvement and be a new indicator of disease activity in Beh(c)et's disease (BD).Methods Thirty-nine consecutive BD patients (who fulfilled the modified International Study Group on BD in 1990 or International Criteria for BD in 2006) and 67 age and sex-matched healthy controls were enrolled (Including 37 patients with hypertension and 30 healthy subjects).The plasma levels of EMPs were measured by flow cytometry utilizing specific labels for endothelial MPs (CD31+ and CD42b-).The measurement data of each group were expressed as-x±s,and the comparison data betwen groups were analyzed by independent sample t test and analysis of variance,Spearman/Pearson correlation analysis,P＜0.05 was statistically significant.Results The levels of circulating EMPs (CD31 + and CI42b-) were significantly elevated in the case group compared with the healthy control group and hypertension (F=6.845,P＜0.05).Moreover,BD patients plasma EMPs were positively correlated with active BD (r=0.802,P＜0.05).Vascular involvement in BD patients was higher than in patients without vascular EMPs,t=4.707,P＜0.05.Gastrointestinal involvement in BD patients was more frequent than that in patients without Gastrointestinal involvement,t=2.673,P＜0.05.Conclusion Levels of circulating EMPs are elevatedd in BD patients and correlated with disease activity in BD.Elevated EMPs may be a potential indicator to predict disease activity of BD.The plasma level of EMPs is increased,which indicats increased risk of vascular and digestive tract involvement in BD.

Objective To evaluate the clinical efficacy of intra-articular injection of ozone in collagen-induced arthritis (CIA) rats and to assess its effects on serum receptor activator of nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) levels. Methods Forty weight age malched Wistar rats were randomly divided into normal control group (normal group), the CIA model group (CIA group), ozone (O 3 group), and methotrexate (MTX group). In addition to the normal control group, Freund's complete adjuvant and bovine type Ⅱ collagen were injected to establish the rat model of CIA. After the model was sucessfully developed, double ankle injection concentration ozone group of 40 μg/ml of O3 each 1 ml, 1 times a week, a total of injection for 3 weeks for the experimenal group. MTX group of 0.9 mg/kg was injected 1 times a week for 3 weeks for the MTX group. Degree of foot swelling was measured, and radiographic assessment of arthritis index (AI) score was performed. One week after treatment, angular vein blood was collected for the rats after the intervention, flow multi-factor detection technology was used to test each rat. T test or Wilcoxon rank test was used to compare the difference between groups. Results ① After 3 week administration with O3, dcgree of foot swelling, and AI of the O3 group was reduced significanly than the CIA group during the same period (foot swelling degree: O3 group: (4.21±0.14) ml, CIA group (9.12±0.17) ml, T=64.08, P=0.00; AI O3 group: [(2.97± 0.18) ml, CIA group: 5.76 ±0.13, T=37.24, P=0.00], and X-ray showed joint damage was alleviated. ② The serum level of RANKL in the CIA group was significantly higher than of the normal group [CIA model group 1890.70(797.03, 10571.94)], normal group [74.46(29.21, 95.37), T=43, P=0.005] during the same period; The serum level of RANKL in the O3 group was significantly lower than the CIA group [O3 group 28.09 (14.11, 207.30), CIA group 1890.70 (797.03, 10571.94), T=39, P<0.05]; RANKL level of the Ozone group when compared with MTX group, was not statistically significantly difference (T=52, P>0.05).③Serum RANKL/OPG of the CIA group was significantly higher than that of the normal group during this period, the difference was statistically significant [CIA group 250.68(42.33, 2959.78), normal group 4.32(3.16,5.30), T=36, P<0.01);The serum level of RANKL/OPG in the O3 group was significantly lower than the CIA group [O3 treatment group 5.10 (3.38, 6.64), CIA group 250.68 (42,33, 2959.78), T=36, P<0.01]; There was no statistically significant difference of the serum RANKL/OPG between the O3 group and the MTX group (T=62.5, P>0.05). Conclusion Intra-articular injection of concentration of 40 μg/ml of O3 can reduce RA rat joint swelling degree, which may relate to the mechanism that O3 can lower levels of serum RANKL and RANKL/OPG ratio, reduce osteoclast formation and activation.

Objective To observe the expression of AdipoQ in Sj?gren's syndrome (SS) mice and its role in inflammation was investigated by recombinant gene transfection study in vivo. Methods As spon-taneous SS mice model, a total of 30 NOD mice were divided into 3 groups randomly: recombinant adenovirus (rADV-AdipoQ) group, normal saline control and simple adenoviruses (control group). The submandibular gland morphology, histopathological grading and the level of serum tumor necrosis factor-α (TNF-α), AdipoQ was compared between the three groups. The expression of AdipoQ on mice submandibular gland was assessed by means of semi-quantitative reverse transcriptase polymerase chain reaction. Results The submandibular glands of the mice of the control group were destructed, with focal lymphocytic infiltration and acinus atrophy. Compared with control model groups, the serum TNF-α and salivary gland AdipoQ expression was significantly down-regulated in the rADV-AdipoQ group [(248 ±30) vs (162 ±73) ng/ml] (P<0.05). Conclusion AdipoQ gene transfected SS mice can significantly improve the morphological features of tissues and decrease the concentration of TNF-αin serum, in addition, it can effectively inhibit inflammation, decrease the degree of protein and AdipoQgene expression. So the AdipoQ may be the protective gene in SS mice.

Objective To assess the improvement of articular symptoms on collagen induced arthritis (CIA) rats after injecting arsenic trioxide (ATO) intraperitoneal and observe its effects on serum RANKL, OPG on CIA rats and discuss the possible mechanism on RANK/RANKL/OPG system.Methods Numberd twentyeight Wistar rats with correspond weight and age consecutively, then using random number table select 8 rats as blank control group A.Another other 20 as model group (16 wistar rats are established as CIA models by immunized twice with bovine type Ⅱ collagen and Freund's complete adjuvant emulsion).Using ranllm numbor table divided CIA rast into the CIA model control group B (n=8) and ATO treatment group C (n=8) randomly.After grouping for 3 days, ATO treatment group were injected with ATO liquid intraperitoneally for 1 week (dose of 4 mg· g-1·d-1, last 1 day raise to 8 mg/kg).Serum were collected after 3 days , while rates in the control group were given same quantity of saline solution with the same method.Arthritis index (AI) and X-ray radiography were assessed for limb joint damage.Flow cytometry (FMC) was used to detect chemokines quantitatively of each rats serum, including Nuclear factor kappa B ligand receptor activation factor/bone protection predominate (RANKL/OPG).According to the data distribution, t test or Wilcoxon test were used to compare the difference between groups.Results ① After administration with ATO for 1 week arthritis index of group C reduced significantly than the CIA model group (2.63±0.92, 6.62±0.91, t=8.73, P＜0.01), and X-ray showed joint damage alleviated.② The serum level of RANKL in the CIA model group was significantly higher than the blank control group [(1 890.70(797.03, 10 571.94) pg/ml, 74.46(29.21, 95.37) pg/ml, T=43, P=0.005];③ The serum level of RANKL in the ATO treatment group was significantly lower than the CIA model group [44.78 (21.41, 79.83), 1 890.70 (797.03, 10 571.94), T=47, P=0.01].④ There was no statistically significant difference of the Serum OPG level between the three groups [16.87(6.91, 17.64), 5.32(3.42, 33.14),14.24(6.96, 21.86) pg/ml, x2=5.078, P=0.166].Conclusion The inflammatory mediators in the process of bone metabolism of CIA rats is disordered , the most significant presentation is the rise of serum RANKL level.Intraperitoneal injection of ATO could improve the arthritis index of CIA rats and its mechanism may be reducing the concentration of serum RANKL/OPG ratio.