Recommendations of diagnosis and treatment of giant cell arteritis and polymyalgia rheumatic in China / 中华内科杂志

Polymyalgia rheumatica (PMR) is a syndrome characterized by pain and morning stiffness in the neck and shoulder and pelvic girdles, as well as raised acute-phase reactants, with or without systemic symptoms, such as fever. Giant cell arteritis (GCA) is a systemic vasculitis of unclear etiology that involves systemic arteries, principally affecting medium- and large-sized arteries with skipped, segmental alterations and granulomatous vasculitis seen on histopathology. In China, epidemiological data describing GCA are still limited; thus, the prevalence might be underestimated. The involvement of vessels in GCA can cause irreversible visual impairment or loss and stroke, which are serious complications. PMR is three times more prevalent than GCA, and other specific diseases should be excluded before the diagnosis is established. PMR symptoms can be present in 40%-60% of patients with GCA. Conversely, GCA can develop in 15% of patients with PMR. Chinese Rheumatology Association, based on the clinical diagnosis and treatment guidelines in 2005, utilizing the experience and guidelines of diagnosis and treatment at home and abroad, formulated this specification to standardize the diagnosis and treatment of GCA and PMR and improve the patient′s prognosis.

Determination of hydroxychloroquine and its metabolites in the breast milk of patients with autoimmune disease / 中华风湿病学杂志

Objective:To determine the concentration of hydroxychloroquine (HCQ) and its active metabolite deethylhydroxychloroquine (DHCQ) in breast milk of lactating patients with autoimmune disease. To observe the safety of hydroxychloroquine in lactation period, and to explore the factors that may affect HCQ and DHCQ concentration in the milk.Methods:Lactating patients with autoimmune disease who have taken HCQ for at least 6 months were included in our study. A new high performance liquid chromatography (HPLC) method was established to detect HCQ and DHCQ levels in breast milk. Milk samples were collected at different time points: before taking the drug (0 hours), and 2 hours, 4 hours, 6 hours after taking the drug. In addition, the genotype of cytochrome CYP3A4*1G, CYP3A5*3 and CYP2D6*10 which were related to HCQ metabolism were tested by dideoxy chain termination method. Visual acuity, hearing and growth status of the patients' infants were followed up on a regular basis. T-test, one-way ANOVA and Pearson's test were used for data analysis. Results:In 15 patients, the average concentration of HCQ and DHCQ in the milk of patients taking 200 mg/d were (520±261) ng/ml and (177±112) ng/ml, respectively. While the average concentration of HCQ and DHCQ in the milk of patients taking 400 mg/d were (1 036±374) ng/ml and (397±271) ng/ml, respectively. The peak of HCQ level for 11 patients was at 4 hour after taking the drug, while the others' were at 2 hour. The breast-fed infants did not show any abnormal symptoms of hearing, vision and growth. However, cytochrome gene polymorphism did not affect the peak of HCQ and DHCQ.Conclusion:The concentration of HCQ and DHCQ in breast milk is positively correlated to the dosage. The peak level of HCQ milk is 4 hours after taking the drug. The levels of HCQ and DHCQ at 6 hours are similar as those in the whole blood. It is suggested that patients who take HCQ can feed 4 hours after taking the drug to reduce the HCQ and its active metabolites being absorbed by infants. However, the impact of HCQ on infant safety and gene polymorphism of CYP on milk concentration among individuals needs to be further verified in large sample studies and long-term follow-up.

Clinical features of patients with idiopathic inflammatory myopathy and interstitial lung disease when complicated with pulmonary infection / 中华风湿病学杂志

Objective:Toanalyze the clinical features of patients with idiopathic inflammatory myopathy (IIM) and interstitial lung disease (ILD) when complicated with pulmonary infection.Methods:Clinical data ofconsecutive IIM patients admitted to our hospital from January 2014 to December 2017 were collected. Patients were divided into two groups: pure IIM-ILD and IIM-ILD with pulmonary infection, and the difference in clinical manifestations and lab test results was compared. The ROC curve was used to evaluate the predictive diagnostic value of T lymphocytes. The data was analyzed by the Chi-square test, Mann-Whitney U test and multiple logistic regression analysis. Results:Totally 153 patients were included, in which 51 cases were complicated with pulmonary infection. The incidence of myalgia, rash, cough/expectoration and fever and the levels of aspartate aminotransferase, lactate dehydrogenase (LDH), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were increased in the infection group, while the number of lymphocytes was significantly decreased ( P<0.05). Multivariate logistic regression analysis showed that myalgia, cough/expectoration, serum LDH level >350 U/L and peripheral blood lymphocyte count <0.9×10 9/L were independent risk factors for pulmonary infection in IIM-ILD patients (ORvalues 4.31, 3.81, 2.70 and 2.44, respectively, P<0.05) . Meanwhile, the number of natural killer cells as well as CD3 +, CD3 +CD4 +, CD3 +CD8 + T cells in the infection group was significantly lower than that in the pure IIM-ILD group ( Z values -2.28, -3.094, -2.918, and -2.308, respectively, P<0.05). ROC curve showed that CD3 + T cells combined with CRP improved the diagnostic sensitivity of pulmonary infection in IIM-ILD. Conclusion:IIM-ILD patients are more likely to have myalgia,cough/expectoration as well as increased LDH level when complicated with infection. The decrease in peripheral T lymphocyte numbers may indicate an increased risk of pulmonary infection in these patients.

The effect of cyclophosphamide treatment on the prognosis of patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To evaluate the effect of cyclophosphamide (CTX) on the prognosis of different types of systemic lupus erythematosus (SLE).Methods A retrospective cohort study,in which 1 372 lupus patients were included,was conduted in 26 hospitals in Jiangsu province from 1999 to 2009.The data was analyzed by the chi-square test,Mann-Whitney U test,Kaplan-Meier survival analysis and Cox regression analysis.Results 1 372 SLE patients were followed up,in which 582 patients (42.4％) were treated with cycloph-osphamide and 790 patients (57.6％) were not.There were statistically significant differences in SLE disease activity index (SLEDAI) at admission and discharge between the two groups (t=3.398,P=0.001;t=5.044,P＜0.01).The survival rate was significantly higher in CTX treatment group compared to those without CTX administration (x2=4.667,P=0.031).Female (P=0.018,HR=0.68),disease duration ≤7 years (P=0.033,R=0.697),decreased levels of complement C3 (P=0.047,HR=0.7),and patients treated with cyclophosphamide and glucocorti-costeroid (P=0.047,HR=0.734) were predictors for good prognosis for CTX treatment.In patients with central nervous system (x2=6.611,P=0.010),heart and lung (x2=7.223,P=0.007) and renal system involvement (x2=5.217,P=0.022),there were statistically significant differences in survival rate and survival time between the CTX treated group and untreated group.Conclusion Cyclophosphamide treatment was beneficial for female SLE patients with low complement,short disease duration and important organ involvement such as heart,lung,kidney and central nervous system.

Central nervous system involvement in primary Sj(o)gren's syndrome: a study of 38 patients / 中华风湿病学杂志

Objective To analyze the clinical profile of primary Sj(o)gren's syndrome (pSS) patientswith central nervous system (CNS) involvement.Methods Thirty-eight pSS-CNS patients at Nanjing Drum Tower Hospital between 2012 and 2016 were enrolled.These patients were divided into high activity group and moderate activity group,according to European League against Rheumatism Sj(o)gren's syndrome Disease Activity Index (ESSDAI).The imaging characteristics,clinical features,laboratory examinations and treatment of 38 cases were retrospectively analyzed.Quantitative differences were analyzed by the student's t-test and qualitative data were analyzed with chi-square or Fisher's exact test.Results The prevalence of central nervous system involvement in pSS patients was 2.93％(38/1 296),while 32％(12/38) as the initial symptom of pSS.Recurrence rate was 39％(15/38).Limb weakness,speech difficulty,sensory disorders,blurred visionwere the most frequent symptoms in pSS-CNS.Multiple lesions in Magnetic resonance imaging (MRI) examination and cerebrospinal fluid abnormality were seen in 94％ (15/16) pSS-CNS patients,among which intra-thecal IgG level increased in 50％ (8/16).In addition,the frequencies of lung involvement,immune associated thrombocytopenia,high-titer antinuclear antibody (ANA) were significantly higher in high activity group of pSS-CNS than those of moderate activity group.The value of above items was 4.7,5.0 and 5.3,respectively,and all the differences were significant (P＜0.05).After high-dose corticosteroids and immunosuppressive therapy,61％ (23/38) patient improved,37％(14/38) were unresponsive to treatment,and 3％(1/38) died because of acute massive cerebral infarction.For those unresponsive patients,mesenchymal stem cell transplantation or immune adsorption treatment might be effective.Conclusion The clinical manifestations of central nervous system are diverse,may be the initial presentations in some pSS patients,with low morbidity and high recurrence rate.Image and lumbar puncture are important for diagnosis.Pulmonary involvement,immune thrombocyto-penia,and high-titer ANA are frequently associated with the activity of pSS-CNS.Most patients have good response to the treatment regimens of high-dose corticosteroids and immunosuppressive therapy,mesenchymal stem cell transplantation or immuno-sorption therapy may be considered in those unresponsive cases.

Umbilical cord mesenchymal stem cell transplantation modulated Th17/Treg balance and delayed lupus progression in MRL/lpr mice / 中华风湿病学杂志

Objective To explore the preventive effect of early umbilical cord mesenchymal stem cells (UC-MSCs) transplantation on MRL/lpr mice and the underly mechanisms.Methods Fourteen 10-week-old MRL/lpr mice were labeled and numbered.They were randomly divided into 2 groups by using random number table and injected with 1 ×106 UC-MSCs or PBS via tail vein respectively.Proteinuria was measured with Bradford method every 4 weeks.All mice were sacrificed at the age of 28 weeks, with the level of serum antidsDNA antibody and IL-17 detected by enzyme linked immunosorbent assay (ELISA).Splenic Th17 cells, as well as regulatory T cells (Treg) were examined by flow cytometry.Data were analyzed with t test and Pearson's correlation test.Results The onset of proteinuria was delayed for 4 weeks in UC-MSC-treated group compared with that in the control group.At the age of 28 weeks, the 24 hour proteinuria [(1.78±0.17) mg vs (4.77±0.98)mg, t=2.99, P＜0.05] and the spleen weight [(0.149±0.009) g vs (0.273±0.052) g, t=2.33, P＜0.05] in UC-MSCtreated group were significantly lower than those in the control group.There was also a trend of the decline of serum anti-dsDNA antibody and IL-17 level after UC-MSCs transplantation.Compared with those in the control group, both the percentage and the absolute number of Th17 cells were significantly decreased in UC-MSC-treated group [(0.90±0.19)％ vs (2.81±0.50)％, t=3.54, P＜0.01 and (3.7±0.8)×105 vs (19.3±3.7)×105, t=4.12,P＜0.01].Meanwhile, the percentage of Treg elevated after UC-MSCs treatment.The ratio of Th17/Treg was significantly lower in UC-MSC-treated group than that in the control group (0.11±0.03 vs 0.50±0.09, t=4.23,P＜0.01).Both the ratio of Th17/Treg (r=0.73, P＜0.01;r=0.59, P＜0.05) and serum IL-17 level (r=0.78, P＜0.01;r=0.56, P＜0.05) was positively correlated with the level of 24 hour proteinuria and anti-dsDNA antibody respectively in MRL/lpr mice.Conclusion Early UC-MSCs transplantation helps to delay disease onset and ameliorate disease progression in MRL/lpr mice, which may act through the modulation of Th17/Treg balance.

Mizoribine versus mycophenolate mofetil or intravenous cyclophosphamide for induction treatment of active lupus nephritis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Lupus nephritis (LN) is one of the most serious manifestations of systemic lupus erythematosus. Although there have been substantial improvements in LN treatment over the last decade, the outcome remains unoptimistic in a considerable percentage of patients. The aim of this study was to evaluate the efficacy and safety of mizoribine (MZR), a novel selective inhibitor of inosine monophosphate dehydrogenase, as induction treatment for active LN in comparison with mycophenolate mofetil (MMF) and intravenous cyclophosphamide (CYC).</p><p><b>METHODS</b>Ninety patients with active LN were observed. Thirty patients were given MZR orally at the dose of 300 mg every other day. Thirty patients took MMF at 2 g per day in two divided doses. Thirty patients received CYC intravenously 0.5 g every 2 weeks. Therapeutic effects and adverse events (AEs) were evaluated at the end of 24-week treatment. One-way analysis of variance (ANOVA) followed by Dunn's test was applied to compare the difference among the groups. For comparing categorical data between two groups, χ(2) test was employed.</p><p><b>RESULTS</b>Early responses at week 12 were achieved by 73.3%, 90.0%, and 96.7% in MZR, MMF, and CYC groups, respectively. There was no significant difference in the complete remission rates (22.7%, 24.0%, and 25.0%, respectively) or overall response rates (68.2%, 72.0%, and 75.0%, respectively) among the three groups at week 24. The most prominent drop-down of Systemic Lupus Erythematosus Disease Activity Index scores was observed in MMF or CYC group, and the decline of health assessment questionnaire scores in MZR or MMF group was more prominent than that in the CYC group at week 12. Serum complement 3 (C3) or C4 levels were elevated in all groups after the treatments. CYC was more effective in inhibiting anti-double-stranded DNA antibody, while MZR was more effective in inhibiting antinuclear antibody. The incidences of AEs in patients treated with CYC were significantly higher than those in patients treated with MZR or MMF (24.2% for CYC vs. 3.3% for MZR, and 2.6% for MMF, P = 0.01).</p><p><b>CONCLUSIONS</b>MZR is well tolerated and has an effect similar to MMF in the induction therapy of active LN. MZR may serve as an alternative approach for LN patients.</p>

Effect of sera from SLE patients on senescence of bone marrow-derived mesenchymal stem cells and its mechanism / 中国免疫学杂志

Objective:To explore whether sera from patients with systemic lupus erythematosus affect the senescence of MSCs, and which signaling pathway might be involved in.Methods: Human sera were collected from sex-matched,age-matched active SLE patients (n=6) and healthy volunteers (n=6).Then the complement was inactivated at 56℃ for 30 min.MSCs were cultured in DMEM/F12 containing 10% SLE or normal serum.Three days later MSCs were stained with SA-β-gal.The percentages of SA-β-gal positive cells were evaluated.The mRNA levels of p53 and p21 were detected by Real time PCR.The protein levels of p53/p21 and NF-κB ( p65) were examined by Western blot.Cell proliferation was evaluated by CCK8 assay.Results: Compared with normal serum, MSCs from either healthy controls or SLE patients showed more SA-β-gal positive cells in SLE serum,as well as higher levels of p53/p21.In addition,SLE serum also inhibited the capacity of MSC proliferation.In MSCs treated with SLE serum, the level of NF-κB (p65) protein was significantly upregulated.Conclusion:SLE serum can promote the senescence of bone marrow MSCs,which might be mediated by NF-κB signaling pathway.

Effect of artesunate on OAZ gene expressions in MRL/lpr lupus mice / 中华风湿病学杂志

Objective To investigate the in vivo effects of artesunate (ART) treatment on OAZ gene expressions in MRL/lpr lupus mice.Methods Twenty-four 12-week-old female MRL/lpr lupus mice were randomly divided into two groups by the random number table,i.e.,the lupus control group and the ART treatment group,12 in each group.Besides,another 8 BABL/C mice were recruited as the normal control group.Peripheral blood mononuclear cells (PBMC) were collected from lupus mice before treatment as well as 4 week and 8 week after treatment,and RNA was extracted and reverse transcripted to cDNA.mRNA expression levels of OAZ and Id1-3 were measured by using real-time polymerase chain reaction (PCR),and the data between the two groups were analyzed by t test,a plurality of samples were compared with the single factor analysis of variance.Serum levels of IFN-γ,IL-4,BAFF and ANA were tested by enzyme-linked immunosorbent assay (ELISA).Relationships of the gene expression levels with levels of cytokines and ANA were analyzed.Statistical analysis were uising t test,ANOVA and LSD-t test.Results mRNA expression levels of OAZ,Id1 and Id3 gene (△Ct:12.9±0.8,12.0±0.4,10.2±0.8) in lupus mice were significantly different from 8 weeks after the treatment comparing to those in the lupus control group (△Ct:9.8±1.0,9.3± 1.1,8.1±0.8) and the normal control group (△Ct:13.9±1.2,11.4±0.7,4.7±0.8,10.3±1.0)(F=7.46,P=0.008; F=6.37,P=0.032; F=5.63,P=0.042),and alteration of OAZ mRNA expression levels before and after the treatment were positively correlated with changes of Id1,Id3 levels (r=0.867,0.947; P＜0.05),and levels of IL-4 and ANA were significantly lower 8 weeks after the treatment than those in the lupus control group (P＜0.05); while level of IFN-γwas higher than those in the lupus control group (P＜0.05).Alteration of OAZ mRNA expression levels before and after the treatment were negatively correlated with changes of ANA,BAFF levels (r=-0.955,r=-0.937; P＜0.05) and positively correlated with changes of Th1/Th2 levels (r=0.976,P＜0.01).Conclusion The expression of genes involving in the OAZ and downstream gene are effectively reduced along with the alteration of several cytokines and ANA after ART treatment.OAZ signaling pathway can play an important role in ART treatment for SLE.

Effects of umbilical cord-derived mesenchymal stem cells on matrix metalloproteinases secreted by fibroblast-like synoviocytes from rheumatoid arthritis / 中华风湿病学杂志

Objective To explore the effects of umbilical cord-derived mesenchymal stem cells (UCMSCs) on fibroblast-like synoviocytes(FLSs) from patients with rheumatoid arthritis(RA). Methods collagen-induced arthritis(CIA) models were developed on Wistar rats and 1 ×106 UCMSCs were given by intravenous injection from tail vein on the 17th day. On day 42, rats were sacrificed and synovial tissues were obtained to detect matrix metalloproteinase (MMP)-1, MMP-3 and MMP-13. Synovial tissues from patients with RA and osteoarthritis(OA) treated by knee arthroplasty were used to isolate FLSs. RNA of FLSs were extracted to compare MMP-1, MMP-3 and MMP-13 expression. FLSs and UCMSCs were cocultured through transwell for 72 hours. Then levels of MMPs were compared in the supernatants by Luminex. The MMPs expressed by FLSs and soluble factors expressed by MSCs were detected by real time-polymerase chain reaction(PCR). After adding antibodies to soluble factors, the MMPs expressions of RA FLSs were compared. Results MMP-1 (6.9±5.4, 1.3±1.4, P 22±21, 22±26, P<0.05) expression were inhibited by UCMSCs in vivo. In vitro, MMP-1 (1.8±0.9, 0.9±0.7, t=2.44, P<0.05), MMP-3(2.6±1.7, 1.1±1.0, t=2.25, P<0.05) and MMP-13(2.4±2.3, 0.6±0.7, t=2.37, P<0.05) levels were higher in RA than OA FLSs. After coculture, MMP-13(1.3±1.2, 0.9±1.2, t=3.63, P<0.05) expressed by FLSs were down-regulated, however MMP-1 (1.5±1.4, 6.6±6.0, t=3.90, P<0.05) and MMP-3 (7±17, 22±35, t=2.86, P<0.05) were up-regulated when analyzed by paired t-test. Soluble factors such as I-DO, HGF and IL-10 were elevated. Anti-IL-10 antibody could decrease the function of UCMSCs by inhibiting MMP-13 expression in RA FLSs. Conclusion UCMSCs ameliorates RA by secreting soluble factor IL-10, which may inhibit MMPs expressed by FLSs.

Expression of interleukin 27 in the sera of patients with Sj(o)gren's syndrome and its regulation on interleukin 10 / 中华风湿病学杂志

Objective To measure the level of interleukin (IL)-27 in sera of patients with Sj(o)gren's syndrome (SS) and investigate its role in IL-10 in CD3+CD8-T cells.Methods Serum levels of 1L-27 and IL-10 from 34 SS patients and 33 healthy controls were tested by enzyme linked immunosorbent assay (ELISA).Peripheral blood monocytes (PBMCs) from SS patients were collected and cultured in different conditions (one with PHA,the other with PHA and IL-27).Seventy-two hours later,cells were harvested and the percentages of CD3 +CD8-IL-10+ cells were measured by flow cytometry.Expressions of c-Maf,IL-10 transcription factor,were examined using real-time polymerase chain reaction (PCR).The data were analyzed by t test,Pearson's correlation analysis and multiple linear regression analysis.Results Serum level of IL-27 in patients with SS [(158±34) pg/ml] was significantly higher than that in healthy controls [(139±18) pg/ml,t=2.823,P＜0.01].IL-27 level was positively correlated with IL-10 and IgG levels (r=0.384,P＜0.05; r=0.354,P＜0.05),but negatively correlated with SSDAI (r=0.503,P＜0.01).Multiple regression analysis showed independent correlation of IL-27 with IL-10 and SSDAI in SS patients (β=0.412,P＜0.01; β=-0.525,P＜0.01),but not IgG.CD3+CD8-IL-10+ cells was upregulated by IL-27 in vitro (P＜0.01,n=14),so was c-Maf (P＜0.01,n=22).Conclusion IL-27 could ameliorate disease activity of Sjogren's syndrome.

Effect of interleukin-21 and dexamethasone on follicular helper T cells in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the effect of interleukin(IL)-21 and dexamethasone(Dex)treatment on the percentage of follicular helper T cells(Tfh)in patients with systemic lupus erythematosus (SLE).Methods Peripheral blood mononuclear cells(PBMCs)obtained from 11 SLE patients were cultured in 96 well plates at 106/well with medium as the control,and with rIL-21(200 ng/μl)or rIL-21(200 ng/μl)+Dex(1×10-6 mol/L)for 24 and 72 hours as the study groups.Another 20 samples were collected and incubated with different concentrations of Dex(0,5×10-7 mol/L,1×10-6 mol/L)for 24 hours.After labeling with CD4,PD1 and CXCR5 monoclonal antibodies,the proportions of Tfh cells were assessed by flow cytometry.ELISA was used to detect supernatant antinuclear antibody(ANA)levels.Differences between groups were analyzed by paired t test.Results The percentages of Tfb cells were elevated when incubated with rIL-21 for 24 h and 72 h[24 h(4.3±1.2)％,(5.9±2.4)％;72 h(5.6±4.0)％,(8.0±5.6)％;t=3.48 and 3.05 respectively,P=0.007 and 0.012].Compared with those in the rIL-21 group,the percentages of Tfh cells in the rIL-21+Dex group were decreased(t=4.70 and 2.78,P=0.001 and 0.019).Tfh cells were decreased ina dose-dependent manner when treated with Dex[24 h(4.2±1.3)％;72 h(6.2±4.4)％;t=3.37 and 2.26,P=0.003 and 0.036].There was no difference of supe(rn)atant ANA levels among those groups.Conclusion IL-21 can promote the proliferation of Tfh cells,while Dex inhibits Tfh cell growth and the inhibition effect is dose dependent.Tfh cells may represent a new target for the treatment of SLE.

Decreased number and impaired function of endothelial progenitor cells in gld.apoE-/-C57BL/6 mice / 中华风湿病学杂志

Objective To detect the changes of number and function of endothelial progenitor cells (EPCs) in gld.apoE-/-C57BL/6 mice,and to investigate whether premature atherosclerosis of gld.apoE-/-C57BL/6 mice was mediated by the dysfunction of these EPCs.Methods EPCs were isolated from peripheral blood and bone marrow of four types of C57BL/6 female mice:gld+/+apoE+/+,gld,apoE-/-and gld.apoE-/-.The percentage of EPCs was analyzed by FACS as CD11b-Sca-1+CD309+ cells.The attached cells were stained with DiI labeled acetylated low-density lipoprotein (DiI-ac-LDL) and FITC-labeled Ulex Europaeus agglutinin 1 (FITC-UEA-1) double staining to determine their phagocytic function.The adherent function of EPCs was determined by calculating the number of re-cultured EPCs.The capacity of EPCs to form the cavity structure was detected by calculating the number of the formed vascular-like structure.Results The percentage of circulating EPCs was significantly decreased in the gld.apoE-/-group (0.012±0.002)％ compared to the gld+/+ apoE+/+ group [(0.039±0.005)％,P＜0.01],the gld group [(0.025±0.001)％,P＜0.05],and the apoE-/-group [(0.028±0.002)％,P＜0.01].The percentage of bone marrow derived EPCs was decreased in the gld.apoE-/-group (0.12±0.01)％ compared to the gld+/+apoE+/+ group [(0.42±0.05)％,P＜0.05].The percentage of DiI-acLDL and FITC-UEA-1 double positive cells was decreased in the gld.apoE-/-group [(59.2±2.1)％] compared to the gld+/+apoE+/+ group [(87.5±3.0)％,P＜0.01],and the gld group [(84.2±6.0)％,P＜0.01] ; the adherent function of EPCs was impaired in the gld.apoE-/-group [(50.0±5.3)％] compared to the gld+/+ apoE+/+ group [(86.0±7.3)％,P＜0.01],the gld group [(73.0±1.0)％,P＜0.01],and the apoE-/-group [(65.0±4.0)％,P＜0.05] respectively.The capacity of EPCs to form the cavity structure was decreased in the gld.apoE-/-group (4.0±0.3) compared to the gld+/+apoE+/+ group (12.0±1.4,P＜0.01).Conclusion The number of EPCs is decreased in the gld apoE-/-C57BL/6 mice,the adhesion,phagocytosis and angiogenic function of EPCs in the bone marrow are impaired,which may be one of the causes of lupus with atherosclerosis.

Expression of microRNA on peripheral blood cells in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To study the expression of 2 bone morphogenetic protein (BMP) related microRNA (miR-21,miR-155) in SLE patients,and to analyze their correlation with clinical features.Methods Peripheral blood was obtained from 59 SLE patients and 25 healthy controls,and total RNA was extracted and reverse transcribed into complementary DNA (cDNA).Real-time PCR technique was used to detect gene expression at transcrptional level.Disease activity was determined by SLEDAI score.Patients were divided into different groups based on their manifestations or antibody profiles,then comparison of miRNA expression was carried out using Mann-Whitney test.Results The expression levels of miR-21 and miR-155 were increased in active SLE patients (SLEDAI≥8) as compared to those with mild disease (SLEDAI≤7) or healthy controls (both P＜0.05).Patients with positive anti-dsDNA or lupus nephritis had higher expression levels of miR-21 and miR-155 than those without (P＜0.05).Conclusion The miR-21 and miR-155 expressions are elevated in active SLE patients and are associated with anti-dsDNA and renal involvement.The results suggest that these two miRNAs might play a role in the pathogenesis of SLE disease.

Role of Olf-1/EBF associated zinc finger protein gene in bone marrow mesenchymal stem cells of systemic lupus erythematosus patients / 中华风湿病学杂志

Objective To study the role of OIf-1/EBF associated zinc finger protein (OAZ),a transcription factor encoded by a positional systemic lupus erythematosus (SLE) candidate gene,in the function of mesenchymal stem cells (MSC) of SLE patients by silencing this gene.Methods OAZ mRNA levels of bone marrow MSC obtained from 5 SLE patients and 5 healthy controls were detected by real-time PCR.Bone marrow MSC obtained from 6 SLE patients were incubated with specific siRNAs for 3 days,then cells were harvested for OAZ measurement,Idl-3 and CCL2 mRNA levels were tested by real-time PCR,and levels of CCL2 were detected in culture supernatants using ELISA.Differences between groups were analyzed using t-test or MannWhitney test.Results ① OAZ mRNA levels of bone marrow MSC were significantly elevated in SLE patients (0.013±0.016) compared to healthy controls (0.001±0.000,P=0.009).② After OAZ silencing,the expression levels of OAZ,Id1,Id2 and ld3 mRNA were significantly decreased (△Ct 10.3±0.7,15.2±1.6,8.1±1.4,10.5±0.6 vs 8.7±0.7,14.1±1.2,7.1±1.5,9.8±0.6) (P all ＜0.05).③ Both the expression levels of CCL2 mRNA (△Ct 2.2±1.1 vs 3.0±1.1 ) and the levels of CCL2 protein in culture supernatants [(341±29) pg/ml vs (304±19) pg/ml] were significantly increased in OAZ silencing group comparing to those in the control group (P all ＜0.05).Conclusion OAZ gene expression is significantly elevated in bone marrow MSC of SLE patients.OAZ may affect autoantibody production in SLE patients by regulating CCL2 expression.

Prognostic indicators for systemic lupus erythematosus analyzed by Cox proportional hazards model / 中华风湿病学杂志

Objective To analyze the prognostic factors and causes of death of patients with systemic lupus erythematosus (SLE).Methods A database with 319 patients were developed.They were newly diagnosed SLE in the Department of Rheumatology and Immunology,Affiliated Drum Tower Hospital of Nanjing University Medical School from 1999 to 2009.Normal distribution of measurement data was presented using mean±standard deviation.The skewed distribution of data was described by median(interquartile range).Using the rate or proportions,the character of classification data was also stated.Survival rate of SLE patients over time was studied by the Kaplan-Meier method,and prognostic factors were analyzed by COX proportional hazards model.Results The 5 year and 10-year survival rates was 96.2%, 88.7%, respectively Prognostic factors affecting survival included duration from onset to diagnosis, anemia, white blood cells in urine, low serum albumin,low C4 level,abnormal ECG and ultrasound echocardiography, pulmonary arterial hypertension (PAH) and systemic lupus erythematosus disease activity index (SLEDAI). However, PAH,duration from onset to diagnosis, low serum albumin were the independent poor prognostic factors and the relative risk and 95% confidence interval were 2.419 (1.052-5.564), 1.162 (1.043-1.294), 0.924 (0.873-0.978), respectively. Renal failure, pulmonary hypertension and infection were the main causes of death,followed by multiple organ failure and lupus encephalopathy. Conclusion PAH, duration from onset to diagnosis, low serum albumin are the important factors predicting poor prognosis. Early diagnosis, timely treatment of SLE organ damages and preventing complications are the key factors to improve the prognosis of patients with SLE.

Clinical effect evaluation of allogenic mesenchymal stem cells transplantation for patients with polymyositis/dermatomyositis / 中华风湿病学杂志

Objective To explore the clinical efficacy of allogenic mesenchymal stem cells transplantation (MSCT) in patients with refractory polymyositis/dermatomyositis (PM/DM). Methods Bone marrow derived mesenchymal stem cells (BM-MSC) or umbilical cord derived mesenchymal stem cells (UC-MSC)were infused intravenously in 8 PM/DM patients. The clinical manifestations and laboratory parameters,including serum creatin in kinase (CK) and manual muscle test 8 (MMT8) score, were compared before and after MSCT. Staistically andlyzed by paired t-test. Results The eight patients were followed up for six to twelve months after MSCT. The level of serum CK decreased from (1681±430) to (886±248) U/L one week after MSCT (P＜0.05) and further decreased at week 2 (474±61) U/L, 1 month (293±89) U/L, 3 month (202±70) U/L and 6 month (175±46) U/L, respectively (all P＜0.05). MMT8 score increased to 1 month [(67±3) vs (45±14), P＜0.05], 3 month [(64±10) vs (45±14), P＜0.05], 6 month [(64±4) vs (45±14),P＜0.05] after MSCT. The dosage of glucocorticoid steroid were tapered in all patients 2 weeks after MSCT [(18±6) mg vs (34±15) mg, P＜0.05]. Clinical symptoms of interstitial pneumonia of both patients were relieved after MSCT, which was confirmed by the result of high resolution CT (HRCT) of the lung.The skin ulcers tended to be recovered after the transplantation in one DM patient. All patients did not develop transplantation related complications. Conclusion Allogenic MSCT is an effective and safe approach for the treatment of refractory PM/DM. However, extensive follow-up study is needed for long-term benefit evaluation.

The study on the expression of EBV antibody in patients with systemic lupus erythematosus / 中华风湿病学杂志

objective To study the role of anti-EBV antibody in the development of systemic lupus erythematosus(SLE).Methods Peripheral blood was obtained from 55 SLE patients and 29 normal controls,and anti-EBV-VCA IgG,IgA,IgM antibody was detected by enzyme-linked immunosorbent assay[ELISA).Totat RNA was extracted from peripheral blood of 33 SLE patients and 26 normal controls and then reverse transcribed into complementary DNA.Real-time PCR technique was used to determine gene expressions at the transcription level.Comparison of anti-EBV antibody level and interferon inducible gene expression was made between SLE Datients and normal controls by t-test.The associations between anti-EBV antibody level,lupus activitv and IFN inducible gene expression were analyzed by Spearman's correlation analysis.Results (1)The levels of anti-EBV antibodies in SLE patients were significantly increased as compared to those in the normal controls[IgG(41±6)vs(19±6)U/ml,IgA(15.4±1.8)vs(8.3±2.1)U/ml,IgM(7.8±1.0)vs(3.7±1.2)U/ml]cantly elevated in SLE Datients as compared to normal controls[IFN scores 11±13 vs 0±3](all P0.05).Conclusion Anti-EBV antibody is overproduced in SLE patients but not associated with disease activity as well as the expression level of several major types of I IFN inducible genes,suggesting that EBV infection may not be a major factor mediating the activation of IFN pathway and consequently the exacerbation of SLE.

Effect of mesenchymal stem cells transplantation on OAZ signaling pathway in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To investigate the in vivo effects of allogeneic mesenchymal stem cells transplantation (MSCT) on OAZ signaling pathway in patients with systemic lupus erythematosus (SLE).Methods Isolated and expand human MSCs from bone marrow cells or umbilical cord of healthy donors were infused into SLE patients. Peripheral blood cells were collected from 10 pre-MSCT patients as well as 1 week and 4 week post-MSCT, and RNA was extracted and reverse transcripted to cDNA. mRNA expression levels of OAZ and Id1-3 were measured by using real-time PCR. Serum levels of IL-10, IL-12, IL-21, CCL2 and ANA were tested by ELISA. Relationships of the gene expression levels with levels of cytokines and ANA were analyzed. Results mRNA expression levels of OAZ, Id1 and Id3 gene in patients with SLE were significantly decreased at week 1(△C:12.4±1.1, 9.7±1.9, 9.7±1.9, 2.1±1.0) and at week 4 (△Ct:13.3±1.2, 10.4±1.5,10.8±1.2, 2.1±1.2) after MSCT when compared to those of the pre-MSCT (△Ct:11.0±0.9, 7.4±2.1, 7.8±2.1, 0.1±1.5 respectively, P all＜0.05). Levels of IL-10, IL-21 and ANA were significantly lower 4 week after MSCT than those before (P＜0.05); while level of CCL2 was higher than pre-MSCT (P＜0.05). Cytokines and ANA levels 1 week after MSCT were not differentially changed comparing to those of the pre-MSCT. Alteration of OAZ mRNA expression levels pre- and post-MSCT were negatively correlated with changes of ANA, IL-21levels and positively correlated with changes of IL-12/IL-10 and CCL2 levels. Conclusions The expression of genes involving in the OAZ signaling pathway is effectively reduced along with the alteration of several cytokines and ANA after allogeneic MSCT in SLE patients. OAZ signaling pathway may play an important role in MSCT treatment for SLE.

Increased interferon-inducible gene expression, an useful marker in the diagnosis and disease activity evaluation of systemic lupus erythematosus / 中华风湿病学杂志

Objective To study the role of the expression levels of 5 type Ⅰ interferon (IFN)-inducible genes (LY6E, OAS1, OASL, MX1, and ISG15) in the diagnosis and disease activity evaluation of systemic lupus erythematosus (SLE). Methods Peripheral blood was obtained from 68 SLE patients, 50 patients with other connective tissue diseases and 26 normal controls, and total RNA was extracted and reverse transcribed into complementary DNA. Real-time PCR technique was used to determine gene expressions at transcription level. An IFN score for each individual was calculated according to the expression of 5 1FN genes. Comparisons of gene expression and IFN score were made among groups. The genes expression levels in patients with SLE were analyzed using receiver operative characteristic curve. The association between IFN scores and disease activity, as assessed by the SLEDAI scores and 24 h proteinuria, was analyzed using Spearman correlation analyses. Results ① The expression levels of MX1, OASL, OAS1, ISG15 and LY6E mRNA in SLE patients were significantly increased as compared with normal controls and disease controls (P all＜0.01 ).② IFN scores in SLE patients (17.9±29.1) were significantly increased as compared with normal controls (0±3.3)and disease controls (3.0±8.1) (P all＜0.01 ). ③ IFN scores area under the ROC curve (AUCROC) was 0.846. When The IFN scores reached 2.56, its sensitivity and specificity for the diagnosis of SLE were 93.1%and 78.3%, respectively. ④ Levels of IFN score was positively correlated with SLEDAI scores (r=0.256,P＜0.05) and 24 h proteinuria (r=0.337, P＜0.05). Conclusion The 5 IFN-inducible genes are highly expressed in SLE patients. IFN score level is valuable for the diagnosis of SLE and a high IFN score is usually associated with an elevated disease activity.