Expression, localization, and clinical application of the RNA binding domain of U1-70kD in HEp-2 cells.

OBJECTIVE: To develop an improved substrate for indirect immunofluorescent test (IIF) to detect anti-U1-70kD autoantibodies. METHODS: The RNA binding domain of U1-70kD (U1BD) complementary DNA was obtained from human larynx carcinoma cell line HEp-2 by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into the mammalian expression vector pEGFP-C1. The recombinant plasmid pEGFP-U1BD was transfected into HEp-2 cells. Immunoblotting (IBT), confocal fluorescence microscopy, and IIF were used to confirm the expression, localization, and antigenicity of fusion proteins of green fluorescent protein (GFP) in transfected HEp-2 cells, which were then analyzed by IIF using human reference sera and compared with untransfected HEp-2 cells simultaneously. RESULTS: (1) The HEp-U1BD cells thus obtained retained their ability to express U1BD-GFP, which showed the antigenicity of U1BD with a characteristic phenotype in IIF. (2) Fifteen IBT-positive anti-U1-70kD sera presented with characteristic cytoplasmic staining on HEp-U1BD by IIF, but five sera without the 70kD reactive band in IBT were not found in the presence of HEp-U1BD pattern. Ten sera of healthy donors couldn't react with HEp-2 and HEp-U1BD at 1:80 attenuant degrees. (3) No differences in expression, localization, or morphology were observed when HEp-U1BD or HEp-2 interacted with the reference sera that could react with Ro/SSA, La/SSB, centromere, histone, and Scl-70 antigens in routine IIF test. CONCLUSIONS: HEp-U1BD cells kept the immunofluorescent properties of HEp-2 cells in an immunofluorescence anti-nuclear antibody (IFANA) test and could be potentially used as a substrate for routine IFANA detection.

Expression, localization and clinical application of exogenous Smith proteins D1 in gene transfected HEp-2 cells.

AIM: To establish an improved substrate for an indirect immunofluorescence test (IIF) to detect anti-Sm antibody. METHODS: Full-length Smith protein D1(Sm-D1) complementary DNA was obtained from human larynx carcinoma cell line HEp-2 by reverse transcription - polymerase chain reaction (RT-PCR) and cloned into the mammalian expression vector pEGFP-C1. The recombinant plasmid pEGFP-Sm-D1 was transfected into HEp-2 cells. The expression, localization and antigenicity of fusion proteins of green fluorescent protein (GFP) in transfected cells were confirmed by means of immunoblotting (IBT), confocal fluorescence microscopy and IIF analysis. Transfected HEp-2 cells were analyzed with reference serum and compared with untransfected HEp-2 cells by IIF. RESULTS: Stable expression of the Sm-D1-GFP was maintained for more than ten generations. This Sm-D1-GFP showed the antigenicity of Sm-D1 with a characteristic phenotype in IIF.Six of 12 serum specimens from systemic lupus erythematosus contained both 29/28 and 13.5 kDa proteins and showed characteristic immunofluorescent patterns. The same phenomenon appeared in 3/6 serum samples which contained 29/28 kDa proteins only. Sera from 10 healthy donors did not react with HEp-Sm-D1 or HEp-2 at 1:80 attenuant degrees. No alteration in expression, localization and morphology was observed when HEp-Sm-D1 or HEp-2 interacted with the reference sera which could react with Ro/SSA, La/SSB, ß2GP1, centromere, histone, and Scl-70 antibodies in routine IIF tests. CONCLUSION: As a new kind of substrate of IIF, HEp-Sm-D1 can be used to detect anti-Sm antibodies. Transfected HEp-2 cells keep the immunofluorescent property of HEp-2 cells in immunofluorescence anti-nuclear antibody (IFANA) test and could potentially be used as substrate for routine IFANA detection.

Low-dose imatinib in the treatment of severe systemic sclerosis: a case series of six Chinese patients and literature review.

Systemic sclerosis (SSc) is a progressive fibrotic disorder with no legitimate effective treatment. Several clinical trials had investigated imatinib mesylate with a target dose of 400∼600 mg/day on SSc, and the efficacy is controversial with a generally poor tolerability. We herein reported six female Chinese patients with SSc administered with low-dose imatinib (200 mg/day) for a median of 23 months (10∼30 months). Patients displayed a decreased modified Rodman skin scores (mRSS) by a mean of 6.29 points after 6 months of treatment. Three patients who completed 2 years of treatment achieved a reduction of mRSS by 8, 18, and 30.5 points, respectively. Pulmonary function was improved or stabilized in two patients with interstitial lung disease. Severe gastrointestinal involvement in one patient was attenuated in terms of discontinuation of total parenteral nutrition and restoration of the serum albumin level. Imatinib was well-tolerated in general, although there were two severe adverse events: a bone fracture and a cerebral hemorrhage in two individuals. Both the adverse events were probably not directly related to imatinib and were recovered uneventfully. Our limited data, along with the review of the literature, suggested that low-dose imatinib might be effective and better tolerated in severe SSc that deserves further study.

Anti-alpha-internexin autoantibody from neuropsychiatric lupus induce cognitive damage via inhibiting axonal elongation and promote neuron apoptosis.

BACKGROUND: Neuropsychiatric systemic lupus erythematosus (NPSLE) is a major complication for lupus patients, which often leads to cognitive disturbances and memory loss and contributes to a significant patient morbidity and mortality. The presence of anti-neuronal autoantibodies (aAbs) has been identified; as examples, anti-NMDA receptors and anti-Ribsomal P aAbs have been linked to certain pathophysiological features of NPSLE. METHODS AND FINDINGS: In the current study, we used a proteomic approach to identify an intermediate neurofilament alpha-internexin (INA) as a pathogenetically relevant autoantigen in NPSLE. The significance of this finding was then validated in an expanded of a cohort of NPSLE patients (n = 67) and controls (n = 270) by demonstrating that high titers of anti-INA aAb was found in both the serum and cerebrospinal fluid (CSF) of approximately 50% NPSLE. Subsequently, a murine model was developed by INA immunization that resulted in pronounced cognitive dysfunction that mimicked features of NPSLE. Histopathology in affected animals displayed cortical and hippocampal neuron apoptosis. In vitro studies further demonstrated that anti-INA Ab mediated neuronal damage via inhibiting axonal elongation and eventually driving the cells to apoptosis. CONCLUSIONS: Taken together, this study identified a novel anti-neurofilament aAb in NPSLE, and established a hitherto undescribed mechanism of aAb-mediated neuron damage that could have relevance to the pathophysiology of NPSLE.

Evaluation of risk factors that contribute to high prevalence of premature atherosclerosis in Chinese premenopausal systemic lupus erythematosus patients.

OBJECTIVE: To evaluate the prevalence of atherosclerosis in Chinese premenopausal women with systemic lupus erythematosus (SLE) and study possible associations between traditional and nontraditional risk factors with premature atherosclerosis. METHODS: We evaluated 111 premenopausal women with SLE and 40 healthy controls without clinical cardiovascular disease. B-mode ultrasound was used to measure carotid plaque and intima-media wall thickness (IMT). The frequency of risk factors for atherosclerosis in patients and controls was compared, and the relationship between the patients' clinical characteristics and carotid plaque was examined. At the same time, we used B-mode ultrasound to measure flow-mediated dilation (FMD) and nitroglycerin-mediated dilation (NMD) in the brachial artery to assess for difference in endothelial function between SLE patients and controls. RESULTS: Carotid plaque was more frequent in patients with lupus (16 of 111 patients) than in control subjects (0 of 40 subjects) (P = 0.007). The mean IMT was significantly higher in patients than in controls. Compared with controls, SLE patients were found to have a significantly higher prevalence of hypertension (P = 0.001), hypercholesterolemia (P = 0.022), and hypertriglyceridemia (P < 0.001). As compared with patients without plaque, patients with plaque were significantly older, had longer disease duration, higher body mass index, raised blood pressure, shorter prothrombin time, raised C-reactive protein, higher Systemic Lupus International Collaborating Clinics damage index score, higher cumulative prednisone dose, used less hydroxychloroquine, had higher mean IMT, lower FMD, and NMD. In logistic regression analysis, older age, higher body mass index, and higher Systemic Lupus International Collaborating Clinics damage index score were independently related to the presence of plaque. Using multiple regression analysis, we found SLE (P = 0.003) to be significantly associated with impaired FMD. CONCLUSION: In our Chinese SLE group, patients presented a higher prevalence of carotid atherosclerosis plaque than healthy controls. SLE patients have significant endothelial dysfunction. We found that risk factors identified in other SLE populations were associated with atherosclerosis in our Chinese group.

Glycogen storage disease manifested as gout and myopathy: three case reports and literature review.

Glycogen storage diseases (GSDs) are a group of congenital inherited metabolic diseases. They may present the symptoms of muscle and joint which may be misdiagnosed with some rheumatic diseases. We report three cases of GSDs in order to have a more clear recognition of the disease and to discuss the differential diagnosis.

Modulatory effects of 1,25-dihydroxyvitamin D3 on human B cell differentiation.

1,25-Dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) can modulate immune responses, but whether it directly affects B cell function is unknown. Patients with systemic lupus erythematosus, especially those with antinuclear Abs and increased disease activity, had decreased 1,25(OH)(2)D(3) levels, suggesting that vitamin D might play a role in regulating autoantibody production. To address this, we examined the effects of 1,25(OH)(2)D(3) on B cell responses and found that it inhibited the ongoing proliferation of activated B cells and induced their apoptosis, whereas initial cell division was unimpeded. The generation of plasma cells and postswitch memory B cells was significantly inhibited by 1,25(OH)(2)D(3), although the up-regulation of genetic programs involved in B cell differentiation was only modestly affected. B cells expressed mRNAs for proteins involved in vitamin D activity, including 1 alpha-hydroxylase, 24-hydroxylase, and the vitamin D receptor, each of which was regulated by 1,25(OH)(2)D(3) and/or activation. Importantly, 1,25(OH)(2)D(3) up-regulated the expression of p27, but not of p18 and p21, which may be important in regulating the proliferation of activated B cells and their subsequent differentiation. These results indicate that 1,25(OH)(2)D(3) may play an important role in the maintenance of B cell homeostasis and that the correction of vitamin D deficiency may be useful in the treatment of B cell-mediated autoimmune disorders.

Some plasmin-induced antibodies bind to cardiolipin, display lupus anticoagulant activity and induce fetal loss in mice.

The combined presence of anti-phospholipid Ab (aPL), thrombosis, and/or fetal loss is recognized as the antiphospholipid syndrome (APS). aPL include anti-cardiolipin Ab (aCL) and/or lupus anticoagulants (LAC, detected as Ig that prolong certain in vitro phospholipid (PL)-restricted blood clotting tests); both aCL and LAC are the diagnostic Ab for APS. Studies show that aPL represent a heterogeneous group of Ab, which recognize various PL, PL-binding plasma proteins, and/or PL-protein complexes. Recently, we found that five of seven patient-derived IgG monoclonal aCL react with thrombin, activated protein C, and plasmin. All three proteins are trypsin-like serine proteases (SP), and are highly homologous in their catalytic domains. Importantly, among these SP autoantigens, the reactive aCL bind to plasmin with the highest affinity, suggesting that plasmin may serve as a major driving autoantigen for some aCL in approximately 30% of APS patients who are positive for IgG anti-plasmin Ab. To test this hypothesis, we immunized BALB/c mice with human plasmin and analyzed immune sera for aCL activity and reactivity with relevant SP. We found that some immune sera displayed aCL activity and/or bound to test SP. Subsequently, eight mAb were obtained and studied. The results revealed that one mAb displayed the aCL and the LAC activities and induced fetal loss when injected into pregnant mice. Immunohistological analyses of placentas revealed extensive deposits of activated C3 components. Combined, these data demonstrate that plasmin may serve as a driving Ag for some pathogenic aPL.

Adult clinically amyopathic dermatomyositis with rapid progressive interstitial lung disease: a retrospective cohort study.

The aim of the study was to investigate the characteristics of adult clinically amyopathic dermatomyositis (CADM) with rapid progressive interstitial lung disease (ILD). Hospitalized patients with dermatomyositis (DM) and polymyositis (PM) between 1998 and 2005 in the Shanghai Renji Hospital were retrospectively studied. One hundred and forty-five patients were classified into CADM, classic DM or PM according to the modified Sontheimer's definition or Bohan-Peter's classification criteria. They were further stratified based on the presence or absence of clinical ILD. The Kaplan-Meier survival analysis and COX regression were performed. The predictive factors for ILD and other clinical properties of CADM-ILD were explored. The presence of clinical ILD was a significant risk factor for the poor outcome of DM/PM (OR = 4.237, CI 95%: 1.239-14.49, p = 0.021). Other risk factors are the presence of rashes and elevated urea nitrogen. Patients with DM/PM complicated by ILD had different clinical courses. Patients with CADM-ILD showed a rapidly progressive pattern with 6-month survival rate of 40.8%. The DM-ILD manifested a progressive pattern with a 5-year survival rate of 54%, while PM-ILD was chronic with 5- and 10-year survival rate of 72.4% and 60.3%, respectively. Better preserved muscle strength, elevated erythrocyte sedimentation rate, and hypoalbuminemia may herald ILD in DM/PM. Patients with CADM-ILD who later died had lower PO(2), higher lactate dehydrogenase, and prominent arthritis/arthralgia compared with those who survived. The presence of antinuclear antibody seems to be protective. Rapid progressive CADM-ILD is refractory to conventional treatment. ILD is a common complication in over 40% of our hospitalized DM/PM cohort and is also a prominent prognostic indicator. CADM is a special phenotype of DM/PM. CADM-ILD, which is usually rapidly progressive and fatal, requires further investigation.

Takayasu's arteritis presenting with bilateral pulmonary granulomatosis.

Takayasu's arteritis (TA) is a vasculitis characterized by inflammation and obliteration of intermediate to large-size arteries. We report a case of Takayasu's arteritis with a presentation of bilateral pulmonary nodular infiltrates in a 21-year-old man. An open-lung biopsy showed characteristic changes of extra-vascular granulomatosis. To our knowledge, this has not been described previously in the literature.

Clinical features, prognostic and risk factors of central nervous system infections in patients with systemic lupus erythematosus.

The purpose of this study is to describe the etiology, characteristics and outcomes of central nervous system (CNS) infections in patients with systemic lupus erythematosus (SLE), while also identifying prognostic and risk factors. Thirty-eight SLE patients with CNS infections were identified from review of all charts of patients with SLE hospitalized from January 1995 to June 2005. These patients were divided into 3 groups, i.e., Mycobacterium tuberculosis (TB), non-TB bacterial and fungal infection groups. Of the 38 SLE cases with CNS infections, TB was identified in 19 patients, Listeria monocytogenes in 3 patients, Klebsiella pneumoniae in 1 patient, Staphylococcus aureus in 1 patient, Gram's stain positive bacteria in 1 patient, Cryptococcus neoformans in 12 patients, and Aspergillus fumigatus in 1 patient. The rate of unfavorable outcome in patients with fungal infection was lower than in patients with TB (P=0.028) and non-TB bacterial CNS infections (P=0.046). SLE patients with TB or fungal CNS infections had a more insidious or atypical clinical presentation. Compared to SLE patients without CNS infections, those with CNS infections were more likely to have low serum albumin levels (P=0.048) and have been receiving higher doses of prednisolone at the onset of CNS infection (P=0.015) or higher mean doses of prednisolone within the previous year (P=0.039). In conclusion, low levels of serum albumin and higher doses of received prednisolone are important risk factors for the development of CNS infections in SLE patients.

[Clinical significance of neuro-reactive autoantibodies in neuro-psychiatric systemic lupus erythematosus].

OBJECTIVE: To investigate the clinical significance of specific/non-specific neuro-reactive autoantibodies in the pathogenesis of neuropsychiatric systemic lupus erythematosus (NPSLE). METHODS: Samples of serum and cerebrospinal fluid (CSF) were collected from 25 NPSLE patients, 11 patients with SLE complicated with central nervous system (CNS) infection (control group 1), and 10 SLE patients without CNS infection (non-NPSLE, control group 2)) were detected, Indirect immunofluorescence technique was used to detect the anti-nuclear antibody (ANA) based on the slides with human epithelium cell of laryngocarcinoma of the line Hep-2 or on the slides with cerebellum and peripheral nerve of monkey, he type of immunofluorescence were assayed. RESULTS: The percentages of ANA positive serum of the NPSLE, SLE control 1, and SLE control 2 groups were 92% (23/25), 100% (11/11), and 100% (10/10) respectively without significant differences between any 2 groups (all P > 0.05). The percentage of ANA positive CSF of the NPSLE was 60% (15/25), significantly higher than that of the SLE control group 1 (27%, 3/11, P < 0.01). There was no significant correlation between the titer of serum ANA and the titer of CSF ANA (P > 0.05). The positive rate of neuro-reactive autoantibodies was 40% (10/25), significantly higher than those of the 2 control groups (both 0, both P > 0.001). CONCLUSION: (1) The ANA positivity in the CSF of the NPSLE group is associated with the disease activity and CNS involvement, the higher the titer of autoantibodies the severe the clinical manifestations. (2) The neuro-reactive autoantibodies in the serum may associate with NPSLE with still unknown mechanism.

[Levels of serum interleukin-15 and the expression of T-helper lymphocyte subsets in peripheral blood of children with juvenile rheumatoid arthritis].

OBJECTIVE: To study the changes of serum interleukin-15 (IL-15) levels and the expression of CD4(+)T (T-helper lymphocyte) subsets CD4(+)CD45RA(+) and CD4(+)CD45RO(+) in peripheral blood of children with juvenile rheumatoid arthritis (JRA). METHODS: The serum concentration of IL-15 was detected using ELISA in 39 children with JRA. The expressions of CD4(+)CD45RA(+)T and CD4(+)CD45RO(+)T in peripheral blood were detected by flow cytometry in 24 out of the 39 patients with JRA. Twenty-six age and sex-matched healthy children were used as the Control group. RESULTS: The mean serum IL-15 level in JRA patients was significantly higher than that in controls (1.37 +/- 0.98 pg/mL vs 0.96 +/- 0.41 pg/mL, P <0.05). Among the 39 JRA patients, the serum IL-15 level in 17 patients with systemic JRA increased remarkably (P < 0.01), but not in patients with the other two types of JRA, the oligoarthritis and polyarthritis (n=13, n=9, respectively), compared with that in controls. The mean serum IL-15 level of the JRA patients was significantly reduced after conventional treatment (P < 0.01). The serum IL-15 level in JRA patients positively correlated with white blood cell count (r=0.347, P <0.05) and C reactive protein (r=0.452, P < 0.01) but not with the erythrocyte sedimentation rate. The patients with high serum IL-15 levels (> or = medium level 1.73 pg/mL) had higher expression of CD4(+)CD45RO(+)T than those with low serum IL-15 levels (< medium level) (16.29 +/- 5.46% vs 11.75 +/- 3.15 %, P < 0.05). CONCLUSIONS: The serum IL-15 levels in JRA patients increased significantly. An increased IL-15 level can transform CD45RA into CD45RO in peripheral blood of patients with JRA, and then result in T lymphocyte activation and mediate the immunopathological impairment. IL-15 may be used a marker for the evaluation of severity of JRA.

[Variations within OLF1/EBF-associated zinc finger protein gene confer susceptibility to lupus nephritis in Chinese population].

OBJECTIVE: To observe whether single nucleotide polymorphisms (SNPs) within the OLF1/EBF-associated zinc finger protein (OAZ) gene are associated with lupus nephritis (LN) susceptibility in Chinese population. METHODS: Eight SNPs located around the positive microsatellite marker D16S517 within OAZ gene with relatively high heterozygosity were chosen for genotyping on 184 systemic lupus erythromatosus (SLE) patients, including 101 non-LN patients and 83 LN patients, and 286 normal controls using TaqMan MGB allelic discrimination method. Data were collected by SDS 2.0 software. Haplotypes and their frequencies were constructed and estimated, and linkage disequilibrium analysis between pairs of SNPs was evaluated by calculating the D Prime using Helixtree program. Case-control study was performed between the SLE, LN, and non-LN groups and normal control group. RESULTS: (1) The frequency of SNP rs1344531 T allele was 47.0% in the SLE patients, significantly higher than that in the controls [38.1%,; chi(2) = 7.300, P = 0.008, OR (95% CI) = 1.441 (1.105 - 1.878)], which showed that the frequency of SNP rs1344531 T allele is associated with SLE susceptibility. The genotypic distribution of SNP rs1344531(CC/CT/TT) differed significantly between the SLE patients (25.5%/54.9%/19.6%) and normal controls (38.1%/47.6%/14.3%) (chi(2) = 8.394, P = 0.015). The CC genotype frequency of the SLE patients was 25.5%, significantly lower than that of the normal controls [38.1%; chi(2) = 7.976, P = 0.005, OR (95% CI) = 0.557 (0.370 - 0.838)] (2) The SNP rs1344531 T allele frequency of the SLE patients was 53.0%, significantly higher than that of the normal controls [38.1%; chi(2) = 11.769, P = 0.001, OR (95% CI) = 1.832 (1.293 - 2.596)], which showed an associated between SNP rs1344531 T allele frequency and LN susceptibility. The genotypic distribution of SNP rs1344531 (CC/CT/TT) differed significantly between the LN patients (22.9%/48.2%/28.9%) and the normal controls (38.1%/47.6%/14.3%) (chi(2) = 12.065, P = 0.002). The CC genotype frequency of the LN patients was 22.9%, significantly lower than that of the normal controls (38.1%) [chi(2) = 6.578, P = 0.013, OR (95% CI) = 0.481 (0.274 - 0.848)]. The TT genotype frequency of the LN patients was 28.9%, significantly higher than that of the normal controls (14.3%) [chi(2) = 9.423, P = 0.003, OR (95% CI) = 2.431 (1.363 - 4.334)]. No statistical significance was observed between the non-LN patients and normal controls in TT genotype frequency. (3) The frequencies of haplotypes containing rs1344531:rs1420676-rs1344531(C-T) [chi(2) = 11.731, P = 0.001, OR (95% CI) = 1.867 (1.302 - 2.676)], rs3803665-rs1420676-rs1344531(C-C-T) [chi(2) = 8.876, P = 0.004, OR (95% CI) = 1.772 (1.213 - 2.589)], and rs2292155-rs3803665-rs1420676-rs1344531(C-C-C-T) [chi(2) = 9.962, P = 0.002, OR (95% CI) = 1.915 (1.274 - 2.880)] were all significantly higher in the LN patients in comparison with the normal control group (41.0% versus 27.1%; 33.3% versus 21.8%; and 27.0% versus 16.2%); while the frequencies of other haplotypes: rs1344531-rs2080353(C-A) [chi(2) = 8.06, P = 0.005, OR (95% CI) = 0.603 (0.424 - 0.856)], rs1344531-rs2080353-rs933564 (C-A-G) [chi(2) = 7.929, P = 0.006, OR (95% CI) = 0.602 (0.422 - 0.859)] were significantly lower than those of the normal control group (39.5% versus 52.2%, and 36.6% versus 49.2%), which produced additional support for such association. CONCLUSION: SNP rs1344531 and some haplotypes containing SNP rs1344531 within OAZ are significantly associated with LN susceptibility. Genetic variants of the OAZ gene are involved in the pathogenesis of LN.

[Peripheral blood mRNA and interferon gene expression in systemic lupus erythematosus].

OBJECTIVE: To investigate the expression levels of interferon (IFN)-alpha, -beta, -omega and -gamma genes and proteins in the peripheral blood of patients with systemic lupus erythematosus (SLE), and to evaluate any possible connections between these expression levels with clinical features. METHODS: 144 SLE patients, 27 non-SLE patients with rheumatisms and 59 normal controls were recruited for the research, and all subjects were drawn blood to isolate plasma and elute total RNA. SYBR Green Dye based real-time quantitative PCR method was used to compare the expression levels of 4 IFNs in patients with SLE and those in the controls. The significance for the correlation of these expression levels and disease activity and specificity was studied. RESULTS: (1) mRNA expression levels of all 4 IFNs in SLE patients were remarkably lower than those observed in normal controls (P < 0.01 in all); IFN-alpha expression levels in SLE patients were increased than those observed in non-SLE group (P < 0.01). (2) The expression levels of all 4 IFNs in active SLE patients were similar to those observed in inactive SLE patients (P > 0.05 in all), and expression levels of all 4 IFNs in patients with SLE were not correlated with involvements of kidney, lung, brain, blood. (3) IFN score in SLE patients was remarkably lower than that observed in normal controls (P < 0.01). (4) The expression levels of all 4 IFNs in both SLE patients and normal controls were positively correlated with each other (P < 0.01 in all). (5) Protein levels of IFN-alpha, IFN-beta and IFN-omega in patients with SLE were similar to those observed in normal group (P > 0.05 in all), protein level of IFN-alpha in active SLE group was apparently elevated than in inactive SLE group (P < 0.01), and protein level of IFN-gamma has a trend to increase in SLE group than in normal group. CONCLUSIONS: Decreased expression levels of IFN-alpha, -beta, -omega and -gamma have implicated significance in the determination of SLE disease specificity, of which IFN-alpha is the best. Lower expression level of IFN score has also significance in the determination of SLE disease specificity. Higher protein level of IFN-alpha is helpful to judging SLE disease activity.

Prevalence and risk factors of knee osteoarthritis in Huang-Pu District, Shanghai, China.

OBJECTIVES: A community-based survey on the prevalence of knee osteoarthritis (OA) and associated factors was carried out in Shanghai, People's Republic of China. METHODS: Of 2,305 registered residents >or=40 years of age, 2,093 were examined by interview and questionnaire. One hundred ninety knee pain-positive and 510 knee pain-negative subjects received radiography. RESULTS: Radiographic knee OA (RKOA) was found in 72.1% (137/190) of symptomatic and 41.6% (212/510) of asymptomatic subjects (SKOA and AsKOA, respectively). The estimated prevalences of symptomatic and asymptomatic knee OA in the community were 7.2% and 37.4%, respectively. Women had higher SKOA prevalence than men (9.8% vs 3.7%, P<0.01). Compared with Caucasians, the urban Shanghai population had a higher prevalence of tibiofemoral joint OA. CONCLUSIONS: There was a higher proportion of SKOA than AsKOA (72.1% vs 41.6%), and SKOA was significantly more associated with disability (81.0% vs 26.9%, P<0.01). The prevalence of SKOA increased with age, from 1.3% in the 40-49-year-old age group to 13.2% in the 70+ group. Age, body mass index, and female gender were associated risk factors for knee OA.

[Interferon-inducible genes lymphocyte antigen 6 complex E and tetratricopeptide repeats 1 are correlated with clinical features of patients with systemic lupus erythematosus].

OBJECTIVE: To investigate the expression levels of lymphocyte antigen 6 complex, locus E (LY6E) and interferon-induced protein with tetratricopeptide repeats 1 (IFIT1) genes in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE), and to evaluate the relations between these gene expression levels and disease activity. METHODS: The clinical data of 144 SLE patients, 27 non-SLE patients with rheumatic diseases, and 59 normal controls were collected. The SLE patients were further divided into 2 subgroups: active SLE group (n = 87) and non-active SLE group (n = 57) according to SLEDAI scores. Specimens of peripheral blood were drawn; total RNA was extracted and transcribed into cDNA. Sybr green dye based real-time quantitative PCR method was used to compare the expression levels (indicated as-DeltaDeltaCT value) of LY6E and IFIT1 in patients with SLE and those in the controls. RESULTS: (1) The-DeltaDeltaCT value of LY6E expression level of the SLE patients was 5.4760 +/- 1.9806, significantly higher than those of the non-SLE patients (3.4323 +/- 1.7456) and normal controls (4.5198 +/- 1.6359, both P = 0.001). (2) The-DeltaDeltaCT value of LY6E and IFIT1 mRNA expression of the active SLE patients were 6.1960 +/- 1.7729 and 6.4997 +/- 2.6297 respectively, significantly higher than those observed in the inactive SLE patients (4.3770 +/- 1.7764 and 4.1327 +/- 2.6044 respectively, both P = 0.000). The-DeltaDeltaCT values of LY6E and IFIT1 mRNA of the SLE patients were correlated with the SLEDAI scores, and with the numbers of matched criteria used in the diagnosis of SLE (P < 0.001). CONCLUSION: As IFN-induced genes, elevated expression of LY6E mRNA is specific to diagnosis of SLE. The real time expression levels of LY6E and IFIT1 genes are associated with SLE disease activity. To inhibit the expression of LY6E and IFIT1 may become a novel therapeutic approach for SLE.

[Using GST-tag to capture protein interaction of an interferon-inducible systemic lupus associated gene, IFIT1].

OBJECTIVE: To investigate the protein-to-protein interaction of interferon-induced protein with tetratricopeptide repeats 1 (IFIT1), a newly discovered systemic lupus erythematosus (SLE) related up-regulated gene, and its possible function. METHODS: Peripheral blood of 40 SLE patients was obtained to extract total RNA and synthesized cDNA. Real-time PCR was used to determine the IFIT1 expression at transcript level. Peripheral blood of another 10 SLE patients was extracted to obtain specimens of white blood cell lysate. Molecular cloning and a modified gluthion S-transferase (GST)-pull down assay were used to capture the protein interacting with IFIT1 in the specimens of white blood cell lysate. MALDI-TOF mass spectrometry (MS) was preformed to identify the captured protein that could interact with IFIT1. Twenty-nine sex and age-matched healthy persons were used as controls. RESULTS: By real-time PCR showed that the IFIT1Delta Ct value (x +/- s) was 2.344 +/- 1.200 in the SLE patients and was 3.734 +/- 1.274 in the controls (P < 0.001), showing a significant up-regulation in SLE patients. IFIT1 was cloned and GST-IFIT1 fusion protein was expressed in Escherichia coli. GST-IFIT1 fusion protein was further purified using Glutathione Sepharose 4B column, and was treated as bait to capture prey from peripheral white blood cell lysate of SLE patients. MALDI-TOF MS detected protein interaction between Rho/Rac guanine nucleotide exchange factor and IFIT1. CONCLUSION: IFIT1 may interact with Rho/Rac guanine nucleotide exchange factor, and regulate the activation of Rho/Rac proteins, thus being involved in the pathogenesis of SLE.