objecive To chrify clinical characteristics and outcme of acute/subacute interstitial pneumonia(A/SIP)in patients with dermatomyositis.Methods The elinical data of 10 dermatemyositis patients accompanied with A/SIP who hospitalized in April 2006 to April 2008 were reviewed.Data of 9 dermatomyesitis patients with non-A/SIP interstitial lung diseases treated during the same period were also documented for the comparison.The survival rate of patients wag statistically analyzed.Results Compared with those dermatomyositis patients with non-A/SIP interstitial lung diseases,patients with A/SIP had shorter disease courses and higher incidences of fever,heliotrope rash and ground glass opacity in CT image(P＜0.01or＜0.05).However,the levels of serum creatine kinase tended to be normal.After following up 6 months,only 1 patientwithA/SIP survived(P=0.0001).Logistic regression analysis showed the combination treatment of hormonal,cyclophosphamide and cyclosparine might prolong the survival time(P=0.107).Conclusions A/SIP with dermatomyositis is a fatal disease which needs to be early diagnosed and treated.Patients having dyspnea or breathless in the early stage,especially those with recurrent fever,heliotrope rash and normal serum creatine kinase is predicted to develop A/SIP later.A better outcome may be achieved when treating the patients with stemids plus cyclophosphamide and cyclosporine.

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.

Objective To evaluate the role of rheumatoid factor (RF), anti-cyclic citrullinated peptide antibody (ACCP) and anti-keratin antibody (AKA) in patients with rheumatoid arthritis (RA).Methods Serum levels of RF, ACCP and AKA were examined in 82 RA and 56 non-RA patients and their sensitivity and specificity for the diagnosis of RA were exmined. Statistical analysis was performed to test the association between ACCP/AKA and number of tender joints and swollen joints, ESR, CRP, disease activity score (DAS) or Ritchie's articular index (RAI). Results ROC curve was performed for each single auto-antibody and various combinations of any two of the above antibodies. The area under the ROC curve was over 0.5 (P<0.05). The specificity of ACCP and AKA was 91.1% and 92.9% respectively. RF, ACCP and AKA were all sensitive markers for the diagnosis of RA and the sensitivity could be as high as 95.1% when one of these markers was positive. There were statistical differences in the number of swollen joints, ESR and DAS between ACCP positive group and ACCP negative group (P<0.05) and statistical significant difference was observed in tender joint count, swollen joint count, ESR and DAS between AKA positive and negative groups (P<0.05). Conclusion Combined test of ACCP, RF and AKA is useful in routine RA diagnosis.ACCP and AKA may be clinical markers for predicting disease activity and prognosis.

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 P<0.05),but not correlated with the levels of anti-EBV antibody in patients(all P>0.05).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.

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.

Objective To identify the clinical differences between central nervous system (CNS) infection and neuropsychiatric lupus in patients with systemic lupus erythematosus (SLE). Method Clinical manifestations, lab test results and prognosis of 12 SLE patients complicated with CNS infections, hospitalized in Nanjing Drum Tower Hospital in the past four years, were reviewed and compared with those of 15 concomi-tantly treated patients with central neuropsychiatric lupus (NPL). Two-indenpendent samples t test, Mann-whitney test and Fisher exact test were used for statistical analysis. Results 83% of SLE patients with CNS infections were female and the average disease onset age was (37±4) years. As compared to neuro-psychiatric lupus patients (the control group), those patients with CNS infections (infection group) had lower lupus disease activity (SLEDAI score 14.3±1.6 vs 6.4±1.2, P<0.01) and took higher dose of corticosteroids [average prednisone dose (28.3±2.5) vs (8.4±3.0) mg/d, P<0.01 ] and more immunosuppressives agents (83% vs 33%, P<0.05) before the occurrence of CNS symptoms. Headache and fever were more common in the infection group (100% vs 46.7% and 91.7% vs 20%, both P<0.01) and simultaneously higher serum albumin levels [(34.2±1.2) g/L vs (29.9±1.6) g/L] were detected in those patients compared to the NPL patients (P<0.05). Cerebrospinal fluid examination showed that agents for a long time but without strong evidence of lupus disease activity, CNS infection should be considered at the appearance of headache and fever, and timely cerebrospinal fluid examination is required for the diagnosis.

Objective To explore the role of OAZ gene in the pathogenesis of systemic lupus erythe-matosus (SLE) by using RNA interfering technique. Methods Peripheral blood mononuclear cells (PBMC) from SLE patients were collected. Each sample was equally divided into four groups for cell culture in 96 well plates. Specific siRNA for OAZ and GAPDH were concordantly added to the experimental group and the positive control group, while nonspecific siRNA was added to the negative control group and only culture medium was added to the Mock control group. Cells and supernatants were harvested after culturing for 72 hours, then RNA was extracted and reverse transcripted to cDNA. OAZ, Id1, Id2, Id3, Id4 mRNA expression levels were analyzed by using real-time PCR. Levels of IFN-γ, IL-4, IL-10, IL-12, IL-21, CCL2, ANA in the supernatant were tested by ELISA. Relationships between the expression levels of OAZ mRNA with levels of cytokines and ANA were analyzed. Results OAZ, Id1, Id2, Id3 gene mRNA expression levels (△Ct: 12.5±1.4, 8.9±1.5, 4.3±0.8, 8.04±1.1) in the experimental group were significantly decreased comparing to those in the negative control group (△Ct: 10.2±1.1, 6.5±1.2, 2.4±1.3, 6.2±1.2 respectively, P<0.05). Levels of IFN-γ, IL-10, IL-12, IL-21 and ANA in the experimental group were significantly lower than those in the negative control group (P< 0.05), but level of CCL2 was higher than the negative control group (P<0.05). Difference of OAZ mRNA expression levels (△△Ct) between the experimental group and the negative control group was negatively correlated with changes of ANA, IL-21 levels, but positively correlated with changes of Th1/Th2, CCL2. Conclusion OAZ siRNA can effectively reduce the expression of genes involved in the OAZ signaling pathway in SLE. OAZ may lead to abnormal production of ANA via regulating Id genes and cytokines.

Objective To explore the expression levels of interferon-inducible genes in patients with systemic lupus erythematosus ( SLE) , and to validate these gene expressions as potential biomarkers for the differentiation of disease flare and infection.Methods Peripheral blood was obtained from 48 SLE, 16 rheumatoid arthritis ( RA) patients and 26 normal controls, and total RNA was extracted and reverse transcribed into complementary DNA.Real-time PCR technique was used to determine the gene expressions of MX1, OASL,OAS1, ISG15 and LY6E at transcription level.Univariate logistic regression analysis and multivariate conditional logistic regression model were applied to analyze 5 related factors for infection or activity.Results ( 1 ) The expression levels of MX1, OASL, 0AS1, ISG15, and LY6E mRNA in SLE patients were significantly increased as compared with normal controls ( P all < 0.01 ) , while the expression levels of OASL,OAS1 ,ISG15 and LY6E mRNA in SLE patients were also higher than those in RA patients (P all <0.05 ).(2)There were no significant difference between male and female patients of the 5 gene expression in SLE patients.(3) By logistic regression analysis, ISG15 and LY6E were independent risk factors for active SLE patients (P <0.01) , OASL expression was an independent risk factor for SLE patients with infection ( P = 0.003 ).Conclusion All the 5 interferon inducible genes are highly expressed in SLE patients, in which ISG15 and LY6E are independently associated with disease flare, while OASL may be helpful for the evaluation of infection in SLE patients.

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.

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.