Objective:To investigate the application of cardiac magnetic resonance (CMR) quantitative techniques in evaluating myocardial involvement differences between new onset and longstanding systemic lupus erythematosus (SLE) patients.Methods:From August 2020 to April 2023, 14 new onset and 15 longstanding SLE patients treated at the First Affiliated Hospital of Anhui Medical University were prospectively included as the study group. Additionally, 18 age-, gender-, body surface area-, and body mass index-matched healthy volunteers were included as the control group. Clinical baseline data, electrocardiograms, and CMR results including left ventricular ejection fraction (LVEF), left ventricular end-systolic volume index (LVESVI), left ventricular end-diastolic volume index (LVEDVI), cardiac index (CI), left ventricular stroke volume index (LVSVI), left ventricular mass index (LVMI), myocardial strain, native T 1 values, and T 2 values were collected. One-way analysis of variance (ANOVA) or Kruskal-Wallis H test was used to compare the quantitative parameters among the three groups. Bonferroni correction was applied for pairwise group comparisons. Results:The native T 1 values [1 114.50 (1 089.33, 1 150.39) ms, 1 085.32 (1 051.31, 1 129.75) ms] and T 2 values [(55.9±3.4) ms, (53.3±1.5) ms] of new onset and longstanding SLE patients were higher than those of the healthy control group [native T 1 values 1052.62 (1024.75, 1077.59) ms, H=17.72, P<0.001; T 2 values (51.2±1.3) ms, F=18.70, P<0.001]. The T 2 values of the new onset SLE group was higher than that of the longstanding SLE group ( P<0.05). The LVEDVI[86.87 (80.80, 93.55) ml/m 2], LVSVI [54.63 (50.42, 59.03) ml/m 2], and LVMI [48.39 (41.65, 53.26) g/m 2] of the new onset SLE group were higher than those of the control group [LVEDVI: 71.11 (65.80, 81.28) ml/m 2, Z=3.02, P=0.003; LVSVI: 42.17 (40.36, 51.33) ml/m 2, Z=2.76, P=0.006; LVMI: 38.48 (35.22, 43.83) g/m 2, Z=3.10, P=0.002]. The LVEDVI and LVSVI of the new onset SLE group were also higher than those of the longstanding SLE group [LVEDVI: 73.30 (69.87, 84.71) ml/m 2, Z=1.97, P=0.048; LVSVI: 45.53 (42.28, 50.98) ml/m 2, Z=2.34, P=0.020]. Conclusion:Myocardial involvement is more severe in new onset SLE patients, whereas acute myocardial injury is alleviated in longstanding SLE patients. Therefore, early detection of cardiac involvement in SLE patients is crucial for improving prognosis.

Objective : To investigate the clinical application value of chemokine ligand (CCL)21 and its receptor chemokine(CCR)7 in patients with anti⁃neutrophil cytoplasmic antibody associated vasculitis. Methods : The ser⁃ um levels of CCL21 and its receptor CCR7 were measured by ELISA methods in 102 patients with AASV and 70 healthy controls. At the same time , the correlation among the CCL21 , CCR7 , MPO , PR3 and general laboratory index of TNF⁃α , IL⁃6 , BUN , CRE , C3 , C4 ESR , CRP , BVAS in AASV patients was analyzed. Results : The serum levels of the CCL21 and CCR7 were significantly higher in AASV patients than that in healthy controls[(276. 95 ± 57. 34) vs ( 179. 91 ± 19. 45) ; (85. 85 ± 18. 77) vs (54. 83 ± 18. 18) pg/ml](P < 0. 05) . The serum levels of the CCL21 and CCR7 in MPO positive patients were higher than those in MPO negative patients[(294. 94 ± 58. 15) vs (244. 35 ± 46. 76) ; (90. 64 ± 18. 38) vs (78. 37 ± 13. 44) pg/ml] ( P < 0. 05 ) . Moreover, The levels of the CCL21 and CCR7 were similar to those of TNF⁃α , IL⁃6 , CRE , BUN , CRP , BVAS and MPO in AASV patients, the levels of the CCL21 and CCR7 in AASV patients were positively correlated. The serum levels of the CCL21 and CCR7 were found to be predictive of AASV activity with ROC. Conclusion The serum level of CCL21 and CCR7 in AASV patients increased significantly , especially in MPO positive patients , and related to patients with renal function index and related inflammation index. Serum CCL21 and CCR7 are the influencing factors of BVAS in AASV patients , and have important significance in the diagnosis and judgment of AASV disease.

Objective:To investigate factors associated with the concentration of hydroxychloroquine (HCQ) and its metabolites in peripheral blood of patients with systemic lupus erythematosus (SLE) who were receiving long-term oral HCQ treatment.Methods:SLE patients who had been taking HCQ for more than 3 months were recruited. Clinical characteristics, laboratory test results and SLE disease activity index (SLEDAI) scores were examined. The concentrations of HCQ and its metabolites from peripheral blood were measured by high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). Student's-test and Nonpara-metric tests were used to compare quantitative data, Chi-square and Fisher's exact tests were used to analyze qualitative data. Correlation between the test results was assessed by correlation coefficient. Variables with P values less than 0.05 in univariate analysis were entered into a logistic regression model. Results:In total, 191 SLE patients on long-term HCQ treatment were included in the analysis. Medians of HCQ blood concentrations ([HCQ]), desethylhydroxychloroquine (DHCQ) blood concentrations ([DHCQ]), desethylchloroquine (DCQ) blood concentrations ([DCQ]) and bisdesethylchloroquine (BDCQ) blood concentrations ([BDCQ]) were 523.19 (402.63, 677.88) ng/ml, 291.79 (212.30, 432.51) ng/ml, 49.37 (35.00, 73.05) ng/ml, 21.78(14.37, 52.46) ng/ml respectively. On multivariate analysis, weight-adjusted oral HCQ dose [ OR(95% CI)=1.366 (1.053, 1.772) , P=0.019], the course of hydroxychloroquine [ OR (95% CI) =0.991 (0.984, 0.999), P=0.026], estimated glomerular filtration rate [ OR(95% CI)=0.984 (0.971, 0.997), P=0.014] and platelet count [ OR (95% CI)=1.010 (1.005, 1.015), P<0.001] were associated with [HCQ]. [HCQ], [DCQ], [BDCQ], [BDCQ]/[HCQ] were negatively correlated with estimated glomerular filtration rate (eGFR) ( r=-0.20, P=0.006; r=-0.19, P=0.010; r=-0.26, P<0.001; r=-0.15, P=0.044, respectively) after adjusted for age, course of disease, duration of HCQ treatment and weight adjusted HCQ dosage, [DHCQ]/[HCQ] was negatively correlated with the SLEDAI score ( r=-0.16, P=0.027) when the effects of glucocorticoid was controlled, [BDCQ]/[HCQ] among different renal function levels was statistically significant ( H=12.46, P=0.014). Conclusion:The factors associated with HCQ blood concentrations in SLE patients on long-term oral HCQ treatment are weight-adjusted HCQ dosage, duration of hydroxychloroquine intake and renal function. In addition, [BDCQ] is closely correlated with renal function, [DHCQ] is correlated with SLE disease activity.

Objective:To investigate the expression and clinical significance of neutrophil myeloperoxidase (MPO) in patients with MPO-antibody associated vasculitis (AAV).Methods:Thirty-six newly diagnosed MPO-AAV patients who were hospitalized in the First Affiliated Hospital, Anhui Medical University from July 2018 to June 2021 were enrolled,and 36 age and sex matched healthy subjects were selected as controls. Neutrophil MPO level was detected by flow cytometry (FCM) and MPO mRNA was tested by real time quantitative polymerase chain reaction (RT-qPCR) in all subjects. Serum complement fragment C5 (C5a) and MPO in both groups and serum MPO-anti-antineutrophilic cytoplasmic antibody(ANCA) in MPO-AAV group were measured by enzyme linked immunosorbent assay (ELISA), while the disease activity was evaluated by Birmingham vasculitis activity score-V3 (BVAS-V3).Results:Compared with the heathy control group, the expression of MPO mRNA in neutrophils, serum MPO and complement C5a in MPO-AAV group were significantly higher[MPO mRNA:30.2±11.5 vs. 1.9±0.6, P<0.001;MPO:(112.0±68.7) IU/L vs. (87.4±22.9) IU/L, P=0.01; C5a:(187.3±90.3) ng/ml vs. (107.3±31.1) ng/ml, P<0.001; respectively], while the mean fluorescence intensity (MFI) of MPO in neutrophils were significantly lower [ 1 343.3±723.4 vs. 2 868.0±1 136.5, P<0.001]. In MPO-AAV group, the expression of neutrophil MPO mRNA was positively correlated with serum MPO-ANCA and MPO levels ( r=0.537, P=0.001 and r=0.358, P=0.032; respectively). Multiple regression analysis suggested that neutrophil MPO mRNA expression was positively correlated with serum MPO-ANCA level ( β=0.695, P=0.006); neutrophil MPO level was negatively correlated with serum MPO-ANCA, MPO and complement C5a levels ( r=-0.335, P=0.046; r=-0.372, P=0.026; r=-0.577, P<0.001; respectively). Further, neutrophil MPO level was negatively correlated with serum complement C5a level ( β=-0.374, P=0.043). BVAS-V3 was positively correlated with MPO mRNA expression in neutrophils, serum MPO-ANCA, MPO and complement C5a ( r=0.598, P<0.001; r=0.599, P<0.001; r=0.537, P=0.001; r=0.415, P=0.012; respectively) and negatively correlated with MPO level in neutrophils ( r=-0.342, P=0.041). In multiple regression analysis it suggested that BVAS-V3 was positively correlated with MPO mRNA expression in neutrophils ( β=0.511, P=0.002). Conclusion:In MPO-AAV patients, MPO synthesis and release in neutrophils are both significantly increased, which might be influenced by serum MPO-ANCA and C5a, respectively. Furthermore, MPO synthesis activity in neutrophils is an independent factor related to disease activity.

Primary biliary cholangitis is a chronic autoimmune cholestatic disease with a progressive course. This disease is not rare in China, but standardized diagnosis and treatment for primary biliary cholangitis are insufficient. Based on the evidence and guidelines from China and other countries, Rheumatology Branch of Chinese Medical Association developed the recommendations of diagnosis and treatment for primary biliary cholangitis in China. The aim is to help clinicians recognize clinical characters, therapeutic selection and prognosis judgement of primary biliary cholangitis, which will contribute to make diagnosis in time, to select treatment properly and to manage follow-up scientifically.

Objective:To investigate the change of peptidylarginine deiminase 4 (PAD4) expression in the neutrophils of peripheral blood of patients with anti-neutrophil cytoplasmic myeloperoxidase antibody-associated vasculitis (MPO-AAV), and to analyze the relationship between the change and disease activity.Methods:Thirty-nine untreated patients with active MPO-AAV (patient group) and thirty-nine healthy volunteers (control group) were enrolled into this study. The PAD4 expressed on neutrophils was detected by flow cytometry (FCM). The serum neutrophil extracellular traps (NETs), fragments from the activated complement C5 (C5a) and anti-neutrophil cytoplasmic myeloperoxidase antibody (MPO-ANCA) were measured by enzyme-linked immuno sorbent assay (ELISA). Their disease activity was evaluated by Birmingham vasculitis activity score (BVAS). All the detected results were compared between the 2 groups by t test, Spearman correlation analysis and multivariate linear regression analysis were employed to analyze the relationship between BVAS and the Lab test results in patient group. Results:The proportion of neutrophil expressing PAD4, the mean fluorescence intensity (MIF) of PAD4, the levels of NETs and C5a in patient group were significantly higher than those in the control group [(71±11)% vs (26±6)%, t=22.456, P<0.01; (33±4) vs (14±4), t=18.668, P<0.01; (0.62±0.22) vs (0.26±0.15), t=8.466, P<0.01 and (4.6±1.0) vs (2.9±1.0), t=7.697, P<0.01, respectively]. In patient group, BVAS was positively associated with the proportion of PAD4 + neutrophil, MFI of PAD4, the serum level of NETs, C5a and MPO-ANCA ( r=0.843, P<0.01; r=0.821, P<0.01; r=0.411 1, P<0.01; r=0.613, P<0.01 and P=0.790, P<0.01, respectively), however, the results of multiple linear regression analysis showed only the percentage of PAD4 + neutrophils and the level of MPO-ANCA were independent influencing factors on BVAS ( β=0.324 6, P<0.01 and β=0.796, P<0.01, respectively). Conclusion:The percentage of neutrophils expressed PAD4 in the peripheral blood of patient with active MPO-AAV is significantly increased, and it is an independent factor affecting the disease activity. Intervention on this expression might be a potential new pathway for MPO-AAV treatment.

OBJECTIVE： To study the relationship of CYP3A4， CYP2C8 and CYP3A5 gene polymorphism with ADR/blood concentration of hydroxychloroquine in patients with autoimmune disease （AID）， and to provide reference for individual medication of hydroxychloroquine. METHODS： Totally 77 AID patients，who were treated with hydroxychloroquine （daily dose of 200 mg to 400 mg） for a long-term （＞6 months）， were selected from the department of rheumatology and immunology， the First Affiliated Hospital of Anhui Medical University during Jul. 2017 to Aug. 2018. The information， blood sample and ADR of them were collected. Those patients were divided into normal liver function group， abnormal liver function group， normal renal function group， abnormal renal function group， normal eye group and abnormal eye group according to the site of ADR. The concentration of hydroxychloroquine was determined by HPLC. Genotype of CYP3A4， CYP2C8 and CYP3A5 were detected by MassARRAY microarray system. The differences of hydroxychloroquine-induced ADR in different genotypes were analyzed by χ2 test. The blood concentration difference of hydroxychloroquine in different genotypes were analyzed by independent sample t-test and one-way ANOVA. RESULTS： There was statistical significance in the distribution of CYP3A5 rs4646453 locus between normal renal function group and abnormal renal function group（P＜0.05）. The incidence of CC genotype was higher than that of AA+AC genotype in abnormal renal function group. There was statistical significance in the distribution of CYP2C8 rs10882526 locus between normal liver function group and abnormal liver function group（P＜0.05）. The incidence of allele G was higher than that of allele A in abnormal liver function group， and the incidence of AG genotype was higher than that of AA genotype. There was no significant correlation of the gene polymorphisms of CYP3A4， CYP2C8 and CYP3A5 with blood concentration among 77 AID patients. In subgroup analysis， blood concentration of GT， GG and TT genotypes of CYP2C8 rs10882521 in 58 patients with systemic lupus erythematosus （SLE） were 514.1，735.3 and 785.9 ng/mL， respectively； GG and TT genotypes were significantly higher than GT genotype （P＜0.05）. CONCLUSIONS： AID patients with CYP3A5 rs4646453 CC genotype have a higher incidence of renal dysfunction due to taking hydroxychloroquine； patients with CYP2C8 rs10882526 locus allele G and AG have a relatively high incidence of renal dysfunction due to taking hydroxychloroquine. When SLE patients taking the same dose of hydroxychloroquine， the blood concentration of hydroxychloroquine in patients carrying CYP2C8 rs10882521 GT genotype is lower than other genotypes.

Objective To investigate the change of CD35 expression on neutrophils in the peripheral blood and the relationship between the change and disease activity in patient with myeloperoxidase antineutrophil cytoplasmic antibody-associated vasculitis (MPO-AAV).Methods Forty untreated patients with active MPO-AAV(patient group)and forty healthy volunteers (control group) were enrolled into this study,and Bermingham vasculitis activity score (BVAS) for every patient was recorded.Flow cytometry (FCM) was employed to detect the CD35 and MPO expression on the neurtrophil,and enzyme linked immunosorbent assay (ELISA) was taken to test the levels of autoantibody against MPO-Antineutrophil cytoplasmic antibody (MPO-ANCA),fragment a from the activated complement factor B (Ba) and MPO in peripheral blood from both group.All test results were compared between the 2 groups by t test,Non-parametric test,Spearman correlation analysis.In addition,the relations among the laboratory results and the relationship between BVAS and the laboratory results were analyzed respectively.Results Compared with the control group,the expression level,which was represented as mean flourscence indensity (MFI),of CD35 and neutrophil membrane MPO on peripheral blood neutrophils was significantly increased [(2 014±968) vs (1 454±511),t=3.024,P=0.002 and (709±244) vs (580±158),t=2.806,P＜0.01,respectively],and the MPO expression level in neutrophils was significantly lower [(1 525±1 033) vs (3 196±2 126),t=-4.468,P＜0.01].Ba and MPO levels in serum of the patient group was significantly higher than that in the control group [37.89(26.17,63.14) μg/L vs 27.99(18.64,46.52) μg/L,Z=-2.521,P=0.012 and 546.16(450.55,729.96) U/L vs 327.93(279.02,365.10) U/L,Z=7.121,P＜0.01,respectively].In patient group,the expression level of CD35 had a significant positive relationship with peripheral blood neutrophil count (r=0.573,P＜0.01),serum Ba (r=0.433,P=0.005) and BVAS (r=0.368,P=0.020),respectively,whereas,there was a negative correlation between the MPO expressed on the neutrophils and that in the neutrophils (r=-0.458,P=0.003),and a positive relationship between MPO-ANCA and BVAS (r=0.351,P=0.026).Conclusion There is significant increased expression of CD35 on the neutrophil of patient with MPO-AAV,which might protect the neutrophil from destruction by the activated complement alternative pathway,and more neutrophils consequently contribute to the MPO-AAV pathogenesis.Inhibition of CD35 expression might become one of the potential new pathways for the treatment of MPO-AAV.

Objective To investigate the change of circulating follicular helper T cells (cTfh) in patients with anti-neutrophil cytoplasmic myeloperoxidase antibody-associated vasculitis (MPO-AAV), and to analyze the relationship between cTfh and disease activity. Methods Thirty-eight untreated MPO-AAV patients (patient group) and thirty-eight healthy volunteers (control group) were enrolled in this study. cTfh and membrane expression of inducible co-stimulator(ICOS)and programmed cell death protein 1(PD-1) were detected by flow cytometry (FCM). Serum anti-neutrophil cytoplasmic myeloperoxidase antibody (MPO-ANCA) was measured by ELISA. Disease activity was evaluated by Birmingham vasculitis activity score (BVAS). Results Compared with those in control group, the proportions of cTfh, ICOS+Tfh and PD-1+Tfh cells in patient group were significantly higher [(25.9±3.8)％vs. (21.0±5.3)％, P<0.001;(1.8±0.8)％vs. (0.8±0.5)％, P<0.001 and (10.2±2.8)％vs. (8.2±2.2)％, P=0.001, respectively]. Meanwhile, the expression of ICOS and PD-1 on cTfh in patient group was markedly more intensive (59.6±10.0 vs.49.2±6.9, P<0.001 and 532.6±104.2 vs. 485.1±73.4, P=0.025, respectively). In patient group, the proportion of cTfh was positively correlated with the ratio of ICOS+Tfh, the expression of ICOS, the level of MPO-ANCA and BVAS (r=0.407, P=0.011; r=0.705, P<0.001; r=0.737, P<0.001 and r=0.663, P<0.001, respectively). The expression intensity of ICOS on cTfh was positively associated with ICOS+Tfh ratio, serum MPO-ANCA and BVAS (r=0.388, P=0.016; r=0.645, P<0.001 and r=0.653, P<0.001, respectively). Nevertheless, the expression of PD-1 on cTfh was only positively correlated with the ratio of PD-1+Tfh (r=0.473, P=0.003). Conclusions Enhanced cTfh in patients with MPO-AAV might produce MPO-ANCA, which is related to the aggravation of MPO-AAV. Thus, cTfh and its ICOS could be potentially targeted for the treatment of MPO-AAV.

Objective To explore preliminarily the role of the E2 subunit of pymvate dehydrogenase (PDC-E2) modified by xenobiotics (e.g.2-octynic acid,2-OA) in the pathogenesis of primary biliary cirrhosis (PBC).Methods Patients of PBC (102 cases),primary sclerosing cholangitis (PSC,34 cases) and healthy controls (HC,50 cases) were selected.The anti-PDC-E2,anti-2-OA and anti-lipoic acid (LA) antibody in the peripheral blood of the 3 groups were tested by enzyme linked immunosorbent assay (ELISA).By inhibitive ELISA (iELISA),30 of the 102 PBC patients with anti-PDC-E2 antibody but without anti-2-OA antibody were selected to detect whether there was any new epitope on the PEC-E2 conjugated with 2-OA.The chi-square test and Fisher exact test were taken to analyze the enumeration data.The two-tailed unpaired t test with Welch's correction was used to compare the measurement data.Spearman rank correlation analysis was also employed for proper test.Results The positive rate of anti-PDC-E2,anti-LA and anti-2-OA antibody in PBC patients was 94.1％(96/102),73.5％(73/102) and 53.9％(55/102) respectively,all of which were statistically significantly higher than those in healthy controls group but were of no significant difference between PSC and healthy controls group.There was no significant relevance between the levels of Anti-LA and anti-2-OA antibody in the PBC group (r=-0.065,P=0.520).The iELISA results showed that the antibody,which only identified the epitopes on 2-OA-PDC-E2 induced by the 2-OA conjugation with PDC-E2,existed in 40％(12/30) of the PBC patients,and more interestingly,this antibody was predominantly appeared in PBC patients at their early clinical stage.Conclusion There are anti-LA antibody and anti-2-OA antibody in PBC patients,which have shown no significant association with each other.It is very likely that new antigenic conformational epitopes on PDC-E2 modified by 2-OA would emerge,which might led to the immune response in the individuals who are susceptible to PBC,and thus contribute for the breaking of PDC-E2 immune tolerance,and PBC occurrence finally.