Objective:Have disclosed that the Behcet's patient have autoantibodies to 160 kD protein which is subsequently identified as protein kinectin by gene library screening using a Behcet's patient serum(BD4).To investigate the antigeneic location of kinectin.Methods:Purified RNA from Hep2 line culture and amplified three kinectin fragment by RT PCR using three pairs of primers(kin 3,base sequence 920～ 1 346;kin M,503～994;kin 5,22～510),which combine to cover nearly the full sequence of kinectin molecules listed in Genbank(Z22551);then cloned the three fragments into pET 42a(+) vector and expressed in BL21(DE3) E.coli.The whole cell lysate was put onto SDS PAGE and subsequently transferred to a nitrocellous membrane,then detected for the antibody of Behcet's patient serum by ECL system.Results:Three PCR fragment posed a 99% correspondent rate with kinectin sequence.All of the expression vectors has a correct readframe and expressed three fusion peptides of molecular weight 89(kin 5)?89(kin M) and 82 kD(kin 3) respectivey by Western blot analysis.Of eight patients,6 patients serum reacted to kin M,5 to kin 3 and 1 to kin 5;none of ten normal controls reacted to all the three fusion fragments.Conclusion:Three PCR fragment of kinectin covering sequence 133 to 4 107(aa22～1 346) have been successfully cloned into pET 42a(+) vector and expressed in BL21(DE3) E.coli.The preliminary analysis demonstrates that the antigeneic region of kinectin is mainly located in the middle and carboxyl terminal portion.

Objective To investigate the susceptible gene loci of SLE.Methods Susceptible loci of chromosome16were found in systemic lupus erythematosus(SLE).According to our previous linkage map-ping and gene chip data,four genes named OAZ,CARD15,DNAJA2and IL-4R were chosen as candidate genes for gene expression research.mRNA extracted from whole blood of42SLE patients and36normal controls were reversely transcipted to cDNA.Then Taqman probe and primers were added to perform real-time PCR in ABI Prism誖7900HT sequence detection system.Housekeeping gene GAPDH was used as a control.Results OAZ and CARD15gene expression was significantly higher in SLE patients than those in normal controls(P

Objective To evaluate the efficacy of medical supervised aerobic exercise on physical activity, quality of life and psychological status in patients with systemic lupus erythematosus (SLE). Methods SLE patients who fulfilled ACR criteria were recruited and divided into 2 groups: exercise group (n=24) and control group (n=25). The patients in the exercise group were supervised to have aerobic exercises. The intensiveness of exercise was determined by 20%-40% of maximum heart rate reservation. Visual analog scale (VAS), SLE disease activity index (SLEDAI) score, physical working capacity (PWC170), SF-36 and profile of mood states (POMS) of the two groups were used to evaluate the changes at the baseline, 1 month, 3 months and 6 months after this study. Results The 2 groups were homogeneous and comparable in disease activity at baseline. 1, 3 and 6 months after the study, the VAS, PWC170, POMS and SF -36 of SLE patients were improved in certain degrees in both groups, while the improvement of VAS (P<0.05), PWCITO (P< 0.01 ) and social function of SF-36 (P<0.05) of exercise group were significantly more evident than those of the control group in 6 months after study without any impact on disease condition. There was a high negative correlation between VAS and 5 categories of POMS (r=-6.26~-0.393, P<0.01 ) and a more relevant positive association between VAS and 2 categories of POMS (r=0.534~0.611, P<0.01). Conclusion The data demonstrate that the supervised aerobic exercise can ameliorate physical capacity, improve quality of life and psychological and emotional status in the state SLE patients without aggravating disease per se.

Objective To investigate the mRNA expression of IKB kinase (IKK-α) and interferon-α (IFN-α) in the peripheral blood leukocytes of patients with systemic lupus erythematosus (SLE), and to explore the role of IKK-α in the production of IFN-α in SLE patients. Methods SYBR green dye I based real-time quantitative PCR was used to detect the mRNA expression levels of IKK-α and IFN-α in the peripheral blood leucocytes of SLE patients and healthy controls. Serum levels of IFN-α were measured with ELISA method. Results IKK-α mRNA expression levels in SLE patients were significantly higher than those of normal controls (P<0.05). IKK-α mRNA expression levels in SLE patients with active disease were significantly higher than patients with stable disease (P<0.01). IFN-α mRNA expression level in SLE patients was significantly lower than that of the normal controls (P<0.01). IFN-α mRNA expression levels in SLE patients with active disease were significantly higher than patients with stable disease (P<0.01). Serum levels of IFN-α in SLE patients with active disease was significantly higher than that of the normal controls and patients with stable disease (P<0.05). The anti-dsDNA antibody correlated positively, and complement C3 correlated negatively with serum concentration of IFN-α. IKK-α mRNA expression levels in SLE patients correlated positively with serum concentration of IFN-α. Conclusion IKK-α correlates positively with serum concentration of IFN-α. The IFN-α level is significantly correlated with disease activity, This suggests that IKK-α may play an important role in the pathogenesis of SLE.

Objective To study 6 type Ⅰ interferon (IFN)-inducible genes (IFIT4, IFI44, Ly6e,OAS1, OAS2 and OAS3) in patients with lupus nephritis (LN) and analyze its correlated expression levels with disease activity and/or clinical manifestations. Methods Total RNA was obtained simultaneously from kidney tissues and peripheral blood cells of 12 patients with diffuse proliferative lupus nephritis and 10 normal controls. Moreover, peripheral blood cells were obtained from 119 LN patients and 35 normal controls. Total RNA was extracted and reversely transcribed into complementary DNA. Gene expression levels were measured by real-time polymerase chain reaction by comparing to a housekeeping gene, and IFN score was calculated. Disease activity was determined by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Results The 6 genes were highly expressed in diffuse proliferative lupus nephritis patients compared with normal controls. IFN scores were positively correlated with SLEDAI score, the concurrent presences of anti-double-stranded DNA (anti-dsDNA) antibodies (P<0.05) and hypocomplementemia (P<0.01). Conclusion The 6 IFN-inducible genes are highly expressed iri LN patients. IFN scores are elevated in active lupus nephritis patients, in patients with positive anti-ds-DNA antibody and hypocomplementemia. IFN scores may be a useful biomarker for lupus nephritis therapy.

0.05).However,Th1was decreased significantly in S LE patients than that in the normal controls(P

Objective To explore the role of chemokines and ehemokine receptors in the etiopathog-enesis of diffuse proliferative lupus nephritis (LN). Methods ① Total RNA from the kidney tissues and peripheral blood cells of 12 patients with diffuse proliferative LN and 10 normal controls were prepared simultaneously and reverse transcribed into complementary DNA. Sybr green dye based real-time quantitative PCR method was used to compare the expression levels (indicated as-AACt value) of MCP-1, CCL19,CXCLg, CXCL10 and CCR2, CCR7, CXCR3. ② Immunofluoresceee labeling and immunohistochemical staining technique were used to observe the distribution of chemokines MCP-1, CCL19, CXCL9 and CXCL10 in normal and patients kidney tissues. Results The 4 chemokines genes (MCP-1, CCL19, CXCL9 and CXCL10) were consistently highly expressed in kidney tissues and peripheral blood ceils of diffuse proliferative LN patients compared with normal controls. The 2 chemokine receptors, CCR2 and CXCR3 were also overexpressed in peripheral blood cells of diffuse proliferative LN patients. There was nearly no expression of these 4 chemokine proteins in normal kidneys. But they were found in glomeruli of diffuse proliferative LN patients. Conclusion The expression of chemokines in the peripheral blood cells may be used as biomarkers for LN. Further study maybe lead to the development of specific drugs targeting at them for the treatment of systemic lupus erythematosus (SLE).

Objective To investigate the pathogenesis of familial systemic lupus erythematosus (SLE), by analyzing the gene expression profile of peripheral blood in a family with 2 SLE patients and their first-degree relatives. Methods Total RNA was extracted from peripheral blood cells of normal subjects and SLE patients. Then, synthesis double strand cDNA template from total RNA, transcription of cRNA probe with Biotin labeling, hybridization of probe with Microarray, binding of Streptavidin to Biotin, amplification with First Antibody, further amplification with Cy3-Conjugated Second Antibody, detection of Cy3 dye with ScanArray 5000 were performed. With QuantArray microarray analysis software, the scan image information was converted into numeric data. With GeneSpring microarray analysis software, cluster analysis was done to find interested genes. Results Over 3000 target genes were analysed. Fifty-nine genes differentially expressed in familial SLE patients and controls were identified. Among them, 34 genes were up-regulated and 25 genes were down-regulated. These differentially expressed genes identified in two familial SLE patients were almost identical to those found in other sporadic SLE patients. Among 34 expression increasing genes, 22 were up-regulated in SLE sisters and unaffected sisters; among 25 expression decreased genes, 17 genes down-regulated in SLE sisters and unaffected sister. Cluster analysis showed that patients were clearly separated from controls and their unaffected sisters based on their gene expression profile. These results showed that in familial SLE, multiple genes were responsible for susceptibility to SLE, and clinically unaffected relatives shared some lupus susceptibility genes with their clinically affected relatives, in addition environmental factors were probably necessary to trigger disease. Conclusion These results indicate that high-density oligonucleotide microarray has the potential to explore the heredity in SLE families.

Objective To continue to study if there is any other pathogenic gene expression related to Th1/Th2 abnormal differentiation,based on the author′s previous results,which have shown that Th1/Th2 unbalance is due to the cytokines and cytokine receptors of differentiation.Methods TaqMan Real time PCR was used to detect the gene expression of Th1/Th2 control in recent onset systemic lupus erythematosus (SLE) patients ( n =38).The genes include I?B,IRF 1,STAT4,GATA3,IL 4R and the others such as CCR1,CCR2,CCR4,CCR5,caepase 1 and CD38,which participate in inflammation,cell apoptosis and so on.Rheumatoid arthritis (RA) patients ( n =50) and normal people ( n =28) were control groups.Results ① Resent onset SLE patients comparing to normal people:STAT4 expression in IL 2/IL 12R ? 2/STAT4 access which induced Th1 differentiation increased significantly ( P 0 05) ;GATA3 expression which induced Th2 differentiation in IL 4/IL 4R/GATA3 access decreased significantly ( P

Objective To explore the pathogenic genes relevant to Behcet's disease (BD) by building the differentail gene expression profiles of peripheral blood leukocytes in BD. Methods Oligonucleotide gene array from Affymetrix Company was applied to study the differed expression levels of whole genome between three age and sex matched BD patients and normal controls. Four genes, BCL6, LRAP, ICOSLG and MME, were selected to be tested for gene expression levels by real-time PCR in the groups of BD, normol controls (NC), Lupus and rheumatoid arthritis (RA) peticnts. Results ① Differential gene expression profile of BD compared to that of normal controls was built up. It contained 89 up-regulated and 57 down-regulated genes. ② Four genes mentioned above had significantly higher expression levels in active BD patients than those in NC but had lower exoression levels in stable BD patients. The expression levels of BCL6 and MME were also proved to be increased significantly in BD than in RA and SLE patients. Conclusion ① Our work shed some light on further research of the etiopathogenesis of BD. ② The expression levels of the four genes are proved to be relevant to BD the first time by us. Further analysis showes that TNF-α and IFN-γ can up-regulate the expression levels of BCL6, LRAP and ICOSLG which may be novel to BD. The MME gene is expressed on the surface of cells, which is convenient for test and may potentially be a marker for the diagnosis of BD.