Objective To compare the difference of peripheral blood adiponectin and CD4+T cell subsets between patients with a-cute gouty arthritis and healthy controls,to explore the correlation between adiponectin and serum uric acid,disease activity,CT4+T cell subsets and some cytokines in patients with gout.Methods The clinical data(including general data,neutrophils,erythrocyte sedimen-tation rate,C-reactive protein,blood uric acid,CT4+T cell subsets and some cytokines)of acute gout group(n=90)and healthy con-trol group(n=72)were collected.The level of adiponectin in peripheral blood of two groups were detected,and the differences of adi-ponectin and CD4+T cell subsets between the two groups were compared;the correlation between adiponectin and clinical data was ana-lyzed.Results The levels of serum adiponectin in the acute gout group were significantly lower than those in the healthy control group(P＜0.001),and the levels of Th2,Thl7,Th17/Treg were significantly higher than those in the healthy control group(P＜0.05),while the levels of Treg and Th1/Th2 were significantly lower than those in the healthy control group(P＜0.05).In the acute gout group,adiponectin was negatively correlated with neutrophil,erythrocyte sedimentation rate and C-reactive proten(r=-0.244,P＜0.05;r=-0.311,P＜0.05;r=-0.506,P＜0.001),there was no correlation with serum uric acid.In acute gout group,adiponectin was posi-tively correlated with Th1 and Th1/Th2(r=0.252,P＜0.05;r=0.218,P＜0.05).In acute gout group,adiponectin in peripheral blood was positively correlated with interleukin-2(IL-2),interleukin-4(IL-4)and interleukin-10(IL-10)(r=0.323,P＜0.05;r=0.377,P＜0.05;r=0.359,P＜0.05).There was a negative correlation between interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)(r=-0.265,P＜0.05;r=-0.299,P＜0.05).Multiple linear regression analysis showed that adiponectin in the acute gout group was negatively correlated with erythrocyte sedimentation rate,C-reactive protein,IL-6 and TNF-α(BESR=-12.541,P=0.003;BCRP=-8.256,P=0.024;BIL-6=-15.907,P=0.037;BTNF-α=-79.770,P=0.040),but positively correlated with Th1(BTh1=2.959,P=0.006).Conclusion The levels of adiponectin in the peripheral blood of patients with acute gouty arthritis were decreased,the levels of Th2 and Th17 were increased,and the levels of Treg were decreased.The decrease of adi-ponectin was related to the immunological disorder and inflammation in the patients with acute gouty arthritis.

Objective:To analyze and summarize the key points of design and implementation of new drug clinical trials for ankylosing spondylitis.Methods:The platform for drug clinical trial registration and information published on the official website of center for drug review and evaluation of national medical products administration (CDE) was searched to obtain data and classified statistics was conducted then. The Mean±SD and M ( Q1, Q3) were used for quantitative data for statistical description, and the rate, composition or relative ratio of qualitative data were used for statistical description. Results:A total of 23 clinical trials meeting the requirements were screened, among which 19 were biological products included in nine phase Ⅲ clinical trials. Among the four chemical drugs, two were phase Ⅱ clinical trials. One of the clinical trials on AS adopted the 1966 New York classification criteria, accounting for 4%. Nineteen of the trials adopted the1984 New York classification criteria, accounting for 83%. Three other trials adopted unspecified classification criteria, accounting for 13%. In one of these clinical trials, the age of patients included was older than 16 years old, 9 trials were 18 to 65 years old, 6 were 18 years old but without upper limit. In the definition of active AS, 19 trials took BASDAI≥4 as the cut-off value for active disease, and BASDAI, total back pain, spinal pain and morning stiffness were regarded as active disease in 4.Conclusion:The number of dosestic AS clinical trial projects continnes to rise. The 1984 classification criteria is adopted as the classification criteria in clinical trials. The minimum age in the inclusion criteria is 18 years old, there is no upper limit in age for inclusion. Disease activity can be evaluated by BASDAI score, combined with comprehensive indicators such as night-time back pain, global spinal pain and morning stiffness.

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.

Systemic lupus erythematosus (SLE) shows a significant gender difference. In addition to X chromosome inactivation (XCI) abnormalities, which may lead to the gender difference in SLE, studies have found that estrogen plays a key role in regulating Th17/Treg immune balance in SLE. Estrogen indirectly affects the quantity and function of Th17 and Treg cells by acting on B cells. In this process, the mutual influence and the interaction between B and T cells promote the development of SLE. Recent studies have reported gender differences in intestinal microbiota, which may lead to sex-dependent genetic susceptibility and epigenetic changes in autoimmune diseases represented by SLE. The interaction between estrogen and intestinal microbiota in SLE affects the immune balance of Th17/Treg cells. This paper mainly reviews the way estrogen works, and the mechanisms by which estrogen regulates Th17/Treg immune balance and the interaction between B and T cells in SLE, hoping to provide new targets for new therapeutic strategies such as selective estrogen receptor modulators.

Cyclic GMP-AMP synthase (cGAS)-stimulator of interferon gene (STING) pathway, as an important part of the innate immune system, is the main pathway for cytoplasmic DNA recognition and cGAS can be triggered by a variety of cytoplasmic dsDNA. This pathway has become an important bridge connecting autoimmunity, aseptic inflammatory response and cell aging. In recent years, cGAS-STING pathway has attracted increasing attention in autoimmune diseases. In rheumatoid arthritis (RA), neutrophil extracellular traps (NETs) induce typeⅠ interferon response and accelerate the production of anti-citrullinated peptide antibody (ACPA) through the cGAS-STING pathway. In addition, the cGAS-STING pathway also participates in synovitis, bone destruction and RA progression by promoting the proliferation and activation of fibroblast-like synovitis cells and the polarization of M1 macrophages. Inhibition of the cGAS-STING pathway or its downstream signaling pathway can reduce synovial inflammation in RA, suggesting that cGAS-STING pathway may be a potential therapeutic target for RA.

Objective:To assess the changes of peripheral blood NK cells in patients with systemic lupus erythematosus (SLE) following belimumab and classic therapy.Methods:From January 2020 to March 2022, peripheral lymphocyte subsets were detected by flow cytometry in SLE patients treated with Belimumab and classic therapy. The duration of treatment was 24 weeks. A total of 40 treated SLE patients were enrolled. The lymphocyte subsets in healthy donors were used as normal control group. Paired sample t-test, rank-sum test, Spearman correlation and generalized linear mixed model were used for statistical analysis. Results:In contrast to healthy subjects, the numbers of NK cells in SLE patients before treatment were significantly decreased [276.0 (179.8, 384.0) cells/μl vs. 61.4 (43.0, 105.1) cells/μl; Z=-7.32, P<0.001], although that after treatment was higher than that before treatment [61.4 (43.0, 105.1) cells/μl vs. 107.7 (72.5, 186.5) cells/μl; Z= -3.22, P<0.001]. Generalized linear mixed model results showed that the increase in serum levels of C3 ( t= -2.94, P=0.006) and NK cells ( t=-2.25, P=0.031) were associated with a decrease of anti-dsDNA antibody titers. The cutoff value of elevated counts of NK cells after treatment was equal or more than 38.5 cells/μl with a sensitivity of 61.9% and a specificity of 81.2%. Compared with those with elevated counts of NK cells ≤38.5 cells/μl, patients with elevated counts of NK cells >38.5 cells/μl had a bigger difference anti-dsDNA antibody [49.2 (0.2, 207.2) vs. 156.2 (19.8, 260.7); Z=-1.55, P=0.120] and a bigger difference of SLEDAI-2000[4.5 (0.0, 10.0) vs. 13.0 (4.5, 18.0); Z=-2.52, P=0.012]. Conclusion:The change in the numbers of NK cells may serve as biomarkers for evaluating the therapeutic responses of SLE. Combinatory approaches employing belimumab and classic therapy may control SLE disease by increasing the number of NK cells

Objective:The common protocol deviation (PD) in clinical trials of drugs for rheumatic diseases were analyzed to provide reference for improving the quality of clinical trials and protecting the safety and rights of subjects.Methods:Protocol deviation (PD) reports of completed projects conducted by Department of Rheumatology and Immunology, the Second Hospital of Shanxi Medical University, from 2017 to 2021 were collected and sorted out for analysis. Results:The common PD and proportion of deviation were deviation from protocol study procedures (25.4%, 44/173), low medication compliance (23.7%, 41/173), visits out of the scheduled visit dates or the tests or examinations out od the inspection window period (19.1%, 33/173), missed visit or missed laboratory tests (17.9%, 31/174), using of prohibited drugs (9.2%, 16/173) and violation of inclusion criteria or exclusion criteria (4.6%, 8/173). 135 cases were mild and 38 cases severe deviation. PPS was eliminated in 5 subjects during statistical analysis.Conclusion:①Investigator training should be strengthened and the operability should be taken into account in clinical trial design. In the implementation process, the inclusion and exclusion criteria should be strictly followed and the should adherent to study procedures. ②The institution should actively promote the development of hospital information system to reduce investigator associated PD. ③The education and management of subject should be strengthened to ensure their compliance and improve the quality of clinical trials to protect the safety and rights of subjects. ④Institute should strengthen the management of trail supervisors and emphasis on problems identification during inspections.

Objective:In this study, the role of IL-17 in the pathogenesis of idiopathic myositis (IIM) was preliminarily investigated by detecting the expression of IL-17 in the muscle tissues of patients with idiopathic inflammatory myositis (IIM) and normal controls.Methods:Twenty-eight patients (20 in DM group with dermatomyositis and 8 in ASS group with anti-synthase syndrome) who were diagnosed with IIM after muscle biopsy and autoantibody detection in our hospital for the first time from October 2019 to August 2021 were included. Twelve cases with normal muscle tissue matched for age and sex were included as the control group. Western blot and immunohistochemical techniques were used to detect the expression level of IL-17 in muscle tissue, and enzyme-linked immunosorbent assay (ELISA) was used to detect serum IL-6. Mann-Whitney U rank sum test was used to compare the difference of IL-17 expression in muscle tissue between the two groups, and non-parametric test was used for comparison between multiple groups. Chi-square test and Spearman rank correlation analysis were used, and P<0.05 was considered statistically significant. Results:① The expression level of IL-17 in IIM muscle tissue[1.63(1.30, 2.05)pg/ml was higher than that in control group[1.00(0.96, 1.00)pg/ml, and the difference was statistically significant ( Z=-3.52, P<0.001). The difference be-tween DM[1.94(1.58, 2.14)pg/ml] and ASS[1.22(1.04,1.55)pg/ml was statistically significant ( Z=-3.20, P=0.001). ② Compared with healthy control group [4.08(3.01, 5.67)pg/ml, the expression of IL-6 in ⅡM serum[8.88(4.93, 13.64) was high ( Z=-3.01, P=0.003), which was positively correlated with the expression of IL-17 ( r=0.42, P=0.027). ③ The ex-pression of IL-17 in muscle tissue was higher in IIM associated with muscle weakness[1.91(1.56, 2.14) pg/ml vs 1.50(1.04, 2.00)pg/ml] ( Z=-1.38, P=0.020), dysphagia [2.06(1.99, 2.14)pg/ml vs 1.62(1.52, 2.04)pg/ml] ( Z=-2.74, P=0.010) and skin involvement[1.98(1.57, 2.14)pg/ml vs 1.04(0.86, 1.61)pg/ml] ( Z=-3.20, P<0.010), and the differences were statistically significant ( P<0.05). ④IL-17 was positively correlated with Myoact-total activity ( r=0.51, P=0.006), Myoact-muscle symptom ( r=0.45, P=0.016), erythrocyte sedimen tation ( r=0.48, P=0.020), and myoenzyme increase ( r=0.56, P=0.002). Conclusion:IL-17 and IL-6 are synergistically involved in the pathogenesis of IIM, suggesting that IL-17 is the therapeutic target of IIM.

Objective:To investigate the level and clinical significance of peripheral blood CD4 +T cell subpopulations in late-onset systemic lupus erythematosus (SLE) patients. Methods:This study included 260 SLE patients hospitalized in the Rheumatology and Immunology Department of the Second Hospital of Shanxi Medical University from January 2016 to December 2021: of whom 58 and 202 were late- (≥50 years) and adult-(18~49 years) onset patients. This study also included 160 subjeces as healthy controls(HCs), of whom 35 and 125 were Control Group 1 (≥50 years) and Control Group 2 (18~49 years). Peripheral blood CD4 +T lymphocyte subsets of these participants were assessed by flow cytometry. The clinical data of all patients and healthy controls (HCs)were recorded. The differences between the groups were analyzed by Mann-Whitney U test or χ2 test. Results:(1)The time of diagnosis of late-onset SLE was longer than that of adult-onset SLE [Median time: 5.0 (2.0, 24.0)months vs 3.0 (1.0, 7.3)months, Z=-3.13, P=0.002]. Compared with adult-onset SLE, the SLEDAI score of late-onset SLE was lower [12.0 (8.0, 15.2) vs 14.0 (10.0, 18.0), Z=-2.12, P=0.034]. Some manifestations occurred more frequently in late-onset SLE, such as weight loss, nausea, abdominal pain, cerebral infarction, interstitial pneumonitis, Sj?gren′s syndrome and infection. The manifestations of skin and mucos a occurred less frequently in late-onset SLE. (2)CD4 +T cell subpopulations: ①The absolute counts of Treg, Th17, Th1 and Th2 cells in the peripheral blood of patients with late-onset SLE were significantly lower than those of HCs [Treg: 10.94 (6.14, 19.23) vs 32.65 (28.07, 41.65), Z=-6.79, P<0.001; Th17: 3.43 (0.94, 5.64) vs 6.13 (3.77, 7.82), Z=-3.24, P=0.001; Th1: 36.02 (10.80, 76.38) vs 128.70(89.82, 159.89), Z=-5.29, P<0.001; Th2:3.56 (1.56, 6.06) vs 8.25 (4.69, 12.98), Z=-4.57, P<0.001]. The ratio of Th17/Treg cells was higher than that of HCs[0.28(0.13, 0.59) vs 0.17 (0.12, 0.28), Z=-2.38, P=0.017].②The absolute counts of Treg, Th17, Th1 and Th2 cells in peripheral blood of patients with adult-onset SLE were significantly lower than those of HCs [Treg: 10.28 (5.37, 17.04) vs.30.19 (21.20, 39.75), Z=-11.28, P<0.001; Th17: 3.44 (1.84, 6.14) vs 6.48 (4.23, 10.66), Z=-6.53, P<0.001; Th1: 29.59(15.14, 56.81) vs 90.75(42.67, 162.00), Z=-7.01, P<0.001; Th2: 2.74 (1.62, 4.77) vs 8.25 (4.75, 11.99), Z=-9.91, P<0.001]. The ratio of Th17/Treg was higher than that of HCs[0.35 (0.17, 0.65) vs 0.23(0.14, 0.37), Z=-3.89, P<0.001].③The ratios of Th17/Treg in patients with late-and adult-onset SLE were higher than those of HCs. The ratio of Th17/Treg was the highest in adult-onset SLE patients. Conclusion:Patients with late-onset SLE have reduced numbers of Treg cells and the immune imbalanced of Th17/Treg. However, the immune imbalance of Th17/Treg in late-onset SLE patients is milder than that in adult-onset SLE patients, which may be related to lower disease activity.

Objective:By detecting the species and distribution of fecal flora in patients with pSS, we investigated the relationship between the alterations of the gut microbiome and its metabolic characteristics with peripheral lymphocyte subsets, and their potential role in the occurrence and development of pSS.Methods:A total of 101 pSS patients who were hospitalized in the department of rheumatology of the Second Hospital of Shanxi Medical University, and 101 age and sex-matched healthy control (HC) in the health check-up center of Shanxi Provincial People′s Hospital from January 2019 to January 2020 were enrolled for 16s rDNA-Amplicon sequencing. The statistical analysis was performed in R software 4.0.3. The Alpha diversity were compared by Wilcoxon test, and Beta diversity were compared by ANOSIM analysis between pSS patients and HC. The difference flora was analyzed by t test. The levels of peripheral lymphocyte subsets pSS patients were detected by flow cytometry. Then the relationship between characteristic flora and clinical indicators such as lymphocyte subsets, erythrocyte sedimentation rate, C-reactive protein, unstimulated whole saliva and stimulated whole saliva were analyzed using Pearson′s correlation analysis. Results:Patients with pSS exhibited a significant reduction in the richness (Chao1, ACE) and diversity (Shannon,Simpson) of gut microbiota compared with those of HC, and there was statistical significant difference in gut microbiota composition (ANOSIM, r=0.09, P=0.001). At the phylum level, the relative abundance of Firmicutes decreased and the relative abundance of Proteobacteria increased in pSS patients. At the genus level, the proportion of Escherichia-shigella ( P<0.001), Lactobacillus ( P<0.001), Bifidobacterium ( P<0.001), Subdoligranulum ( P<0.001), Alistipes ( P<0.001) and [ Eubacterium]_ coprostanoligenes (P=0.002) were increased. The proportion of Faecalibacterium ( P<0.001), Prevotella ( P<0.001), Roseburia ( P<0.001), Megamonas ( P<0.001), Agathobacter ( P<0.001), Lachnospira ( P<0.001), Lachnospira_NK4A136 ( P=0.006), [ Eubacterium] _eligens ( P<0.001) were significantly reduced. PICRUST analysis showed significant enrichment of amino acid metabolism taurine and hypotaurine metabolism ( P<0.001), fatty acid metabolism such as propanoate metabolism ( P<0.001), glutathione metabolism ( P=0.002), lipoic acid metabolism ( P=0.003), lipopolysaccharide biosynthesis and biosynthesis of siderophore group nonribosomal peptides ( P=0.005) and Aminobenzoate degradation ( P=0.002) in patients with pSS. The Pearson correlation results showed that there were significant different in the abundances of the key gut microbiota between the HC and pSS groups, and they were closely related to unstimulated whole saliva, the absolute number of Treg cells and Th17 cells. Conclusion:The dysbiosis and metabolism changes of the pSS intestinal microbiota may contribute to immune homeostasis imbalance, and may be involved in the occurrence and development of pSS.