Clinical manifestations and prognosis of SLE patients with positive anti-cardiolipin antibodies / 医学研究生学报

Objectives Anticardiolipin antibody (aCL) is an important component of antiphospholipid antibody (aPL) and a marker antibody of antiphospholipid syndrome (aPS). APL is positive in 20% to 40% of patients with systemic lupus erythematosus(SLE). This article investigated the clinical features and prognosis of SLE patients with aCL. Methods From January 1999 to December 2009, 495 cases of SLE patients detected aCL who were hospitalized for the first time in 26 hospitals in Jiangsu Province including Nanjing Drum Tower Hospital were divided into aCL-positive group and aCL-negative group according to the test results. The patients were followed up in survival status, and the demographic characteristics, affected organs, laboratory tests, treatment drugs, and prognosis were compared between two groups. Results 146 of the SLE patients in this group were positive for aCL. The proportion of women in aCL- positive group (96.6%) was significantly higher than that in aCL-negative group (90.8%), and the difference was statistically significant (P<0.05). The proportion of anemia (74.7% vs 61.3%), decreased C3(81.5% vs 71.1%), positive antinuclear antibody(97.2% vs 92.4%), and positive anti-dsDNA antibody (61.9% vs 49.6%) in aCL-positive group were significantly higher than those of aCL-negative group, and the difference was statistically significant (P<0.05). The aCL-positive group received a higher proportion of cyclophosphamide immunosuppressive therapy (39.5% vs 50.7%, P<0.05). At the end of follow-up, the mortality rate of aCL-positive group was 13.7%, and the mortality rate of aCL-negative group was 14.9% and there was no significant difference in mortality (P>0.05). Kaplan-Meier survival analysis showed that the 1-year, 5-year, and 10-year survival rates of aCL-positive group were 94.5%, 89.0%, and 82.9%, respectively, and there was no significant difference compared with aCL-negative group(P=0.776). The main causes of death in aCL-positive group were lupus encephalopathy (6 cases, 30.0%), renal failure (5 cases, 25.0%), heart failure (4 cases, 20.0%) and infection (3 cases, 15%). The main causes of death in aCL-negative group were infection (21 cases, 40.4%), lupus encephalopathy (11 cases, 21.2%) and heart failure (5 cases, 9.6%) and renal failure (4 cases, 7.7%). Conclusion SLE patients with aCL represent a high propotion in anemia, decreased C3, positive antinuclear antibody, positive anti-dsDNA antibody. There was no significant difference in disease activity and significant organ involvement between two groups. More SLE patients with aCL were treated with cyclophosphamide, and there was no significant difference in survival status between SLE patients with and without aCL during long-term follow-up.

OAZ gene polymorphism in Chinese patients with systemic lupus erythematosus / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To observe the association between systemic lupus erythematosus (SLE) and gene polymorphisms of OLF-1/EBF associated zinc finger protein(OAZ).</p><p><b>METHODS</b>Verified single nucleotide polymorphisms (SNPs) with relatively high heterozygosity were chosen for allelic discrimination in 244 Chinese SLE pedigrees. Then transmissions of single SNP, and haplotypes were calculated by Genehunter software..OAZ mRNA level was also measured for comparing gene expression in patients of different haplotypes.</p><p><b>RESULTS</b>Genotyping of five SNPs within OAZ gene introns indicated there was no preferential transmission of single SNP, and haplotype T-A-G-G for rs1344531-rs2080353-rs933564-rs1345431 showed only weak linkage with the disease (P=0.04). However, haplotypes combining SNPs and the SLE-associated D16S517 allele showed significant association with SLE susceptibility (for rs933564-d16s517 G-271bp t:non-t=93:29 P<0.000001, for rs2080353-rs933564-d16s517 A-G-271bp t:non-t=88:35 P=0.000002). The haplotype A-G-271bp-G of Rs2080353-rs933564-D16s517-rs1345431 was also transmitted to patients preferentially (P=0.0084) and it showed a tendency to affect gene expression.</p><p><b>CONCLUSION</b>Special polymorphism haplotype of OAZ gene is associated with Chinese SLE. OAZ may suggest a new pathway for lupus.</p>

Single nucleotide polymorphisms of deoxyribonuclease I and their expression in Chinese systemic lupus erythematosus patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Previous studies have suggested that interrupted clearance of nuclear DNA-protein complexes after cell death might initiate and propagate systemic lupus erythematosus (SLE). Deoxyribonuclease I (DNaseI) may be responsible for the removal of DNA from nuclear antigens at sites of high cell turnover, thus preventing the onset of SLE. The purpose of this study was to genotype the single nucleotide polymorphisms (SNPs) of DNase1 and characterize its gene expression and alternatively spliced transcripts in Chinese patients with SLE in order to understand the pathogenic role of DNase1 in human SLE.</p><p><b>METHODS</b>Four SNPs located at the 3' end of the DNase1 gene, as listed on the SNP website, were selected for analysis. Those SNPs with relatively high heterozygosity were chosen for genotyping in 312 Chinese SLE families using the Taqman minor groove binder (MGB) allelic discrimination method. Haplotypes were constructed and linkage disequilibrium tests were performed using GeneHunter. DNase1 mRNA expression was detected using real-time polymerase chain reaction (PCR), and alternatively spliced transcripts were isolated using capillary electrophoresis. Any effects the specific SNP haplotypes had on DNase1 gene expression and the alternatively spliced transcripts were also assessed.</p><p><b>RESULTS</b>rs179982 and rs1053874 had high heterozygosity, about 0.5 in this Chinese cohort, while rs1059857 was also found to be heterozygous. Analysis of the haplotype combining rs179982-rs1030874 (C-G) and rs179982-rs1030874-rs1059857 (C-G-G) revealed a skewed transmission in favor of affected offspring. DNase1 gene expression was higher in SLE patients than in normal controls (P < 0.001), but this was not related to disease activity or SNP haplotype. Capillary electrophoresis revealed that the pattern of alternatively spliced transcripts in patients differed from that of normal controls. Furthermore, different SNP haplotype combinations generated different transcript patterns in SLE patients.</p><p><b>CONCLUSIONS</b>The SNP haplotypes are in linkage disequilibrium in Chinese SLE patients and may induce the disease through a modification of DNase1 mRNA splicing rather than at the level of mRNA expression. There is a relatively unique transcript band in SLE patients independent of special haplotype, which suggests that other unknown factors might be involved in adjusting gene expression.</p>

Deoxyribonuclease I gene expression in systemic lupus erythematosus patients / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To observe whether deoxyribonuclease I (DNASE1) gene expression and its DNASE1 mRNA expression was detected by real-time polymerase chain reaction and its alternatively spliced transcripts were performed by capillary electrophoresis. An analysis was also made to disclose whether specific single nucleotide polymorphisms (SNPs) haplotype had effects onDNASE1 gene expression and its alternatively spliced transcripts.</p><p><b>RESULTS</b>DNASE1 gene expression was higher in SLE patients than in normal controls (P<0.001), and in patients it was found to be of no relationship with SLE disease activity index score. However, it was increased in female patients. Capillary electrophoresis revealed that the pattern of alternatively spliced transcripts in patients was not the same as that in normal controls. Moreover, it seemed that different SNPs haplotype combination might show different transcript pattern in SLE patients.</p><p><b>CONCLUSION</b>In SLE patients, DNASE1 gene expression is abnormal and there are alternatively spliced transcripts different from those in normal controls. DNASE1 gene is a critical factor in the pathogenesis of SLE.</p>

Susceptibility gene of systemic lupus erythematosus in 16q12 in Chinese cohort / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To localize susceptibility loci in Chinese systemic lupus erythematosus (SLE) cohort by linkage disequilibrium mapping the genomic interval in human chromosome 16 so as to understand whether the pathogenesis of human SLE is related to chromosome 16.</p><p><b>METHODS</b>Five microsatellite markers in chromosome 16 spanning from 57.79 cM to 65.1 cM were used for fluorescence based genotyping in 157 SLE families. Evidence for linkage disequilibrium was assessed using the extended transmission disequilibrium test (ETDT) program and Genehunter software. Susceptibility gene was searched in the positive locus, and real-time PCR method was used to detect gene expression.</p><p><b>RESULTS</b>With the ETDT, evidence for linkage disequilibrium of the marker D16S409 (P=0.0278) and D16S517 (P<0.0001) with SLE were found. It was observed that 271 bp allele of D16S517 was much in evidence for preferential transmission, while 277 bp allele was found to be transmitted less often than expected. Genehunter analysis is concordant with ETDT. By real-time PCR, OAZ(OLF1/ EBF-associated zinc finger protein) gene which is located in the positive locus showed higher expression in SLE patients than in normal controls.</p><p><b>CONCLUSION</b>Results reveal that 16q12 (58.46 cM) is associated with Chinese SLE. OAZ is a likely susceptibility gene in this locus for SLE.</p>

Biological characteristics and hematopoietic support of bone marrow-derived mesenchymal stem cells in patients with systemic lupus erythematosus / 中华风湿病学杂志

Objective To explore the biological characteristics and karyotype of bone marrow-derived mesenchymal stem cells(MSCs)in patients with systemic lupus erythematosus(SLE)and hematopoietic sup- port of MSCs.Methods MSCs were isolated from bone marrow of 11 SLE patients and 6 healthy controls by density centrifugation and adhesive culture in vitro.The surface markers were detected by flow cytometry (FCM).The morphological changes of MSCs were observed in primary and passage cultures.The growth curves were assayed.The karyotype of MSCs was detected by blocking cellular mitosis with colchicines.The MSCs from SLE patients and healthy controls were infused to ICR mice after high-dose chemotherapy.The changes of peripheral blood counts of the mice were recorded.Results Approximately(6～9)?10~9 MSCs from SLE were obtained after 5 passages and their growth was slower than normal controls(P＜0.01).Both groups were positive for CD29,CD44 and CD105,and negative for CD14,CD34,CD45 and HLA-DR.MSCs from SLE had a normal karyotype.MSCs infusions of the two groups were accompanied by no adverse event and the recovery of white blood cell,hemoglobin and platelet count was quicker when compared with the controls(P＜0.05).Conclusion MSCs from SLE have demonstrated abnormalities in expansion in vitro.MSCs from SLE have a normal karyotype.Ex vivo MSCs infusion from SLE patients can support hematopoiesis as normal MSCs.