OBJECTIVETo investigate the relationship between haplotypes of multilocus markers and ankylosing spondylitis (AS).

METHODSFive families with AS were recruited from Shanghai area. Eleven microsatellite markers around D6S276 were analyzed by Linkage package and by Cyrillic package.

RESULTSFine linkage analysis showed the significant Lod score values with D6S276 was 3.8821, Lod score values with D6S1691 and D6S1618 near D6S276 were larger than 1.5. The crossover value in 5 pedigrees was 14%. The haplotype analysis showed that the regions between D6S1691 and D6S1618 were associated with AS.

CONCLUSIONThe regions of D6S1691-D6S276-D6S1618 may harbor a susceptible gene of AS. The specific haplotypes of different pedigrees may play an important role in the presymptomatic diagnosis for AS.

Female ; Haplotypes ; genetics ; Humans ; Linkage Disequilibrium ; genetics ; Male ; Pedigree ; Spondylitis, Ankylosing ; genetics

HLA-B Antigens ; genetics ; HLA-B27 Antigen ; genetics ; Humans ; Spondylitis, Ankylosing ; genetics

BACKGROUND: Circular RNA (circRNA) is a type of closed circular noncoding RNA (ncRNA), mostly formed by back-splicing or alternative splicing of pre-messenger RNA (mRNA). The aim of this study was to explore the expression profile of circRNA in peripheral blood mononuclear cells (PBMCs) of patients with ankylosing spondylitis (AS) and discover potential molecular markers of AS. METHODS: The circRNA microarray technology was used to detect the expression of circRNAs in the peripheral blood of 6 patients with AS and 6 healthy controls (HC). To screen the differentially expressed circRNAs by fold change (FC) and P value, these differentially expressed circRNAs were analyzed by bioinformatics. In 60 cases of AS and 30 cases of HC, 4 circRNAs were subjected to real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and their correlation with various clinical indicators was analyzed. Finally, the receiver operating characteristic (ROC) curve was used to analyze their potential as AS diagnostic markers. RESULTS: The microarray results showed that there were 1369 significantly differently expressed (P < 0.05, FC > 1.5) circRNAs between the AS and HC groups (675 upregulated and 694 downregulated). The results of bioinformatics analysis suggested that they were mainly involved in "enzyme binding," "adenosine ribonucleotide binding," "MAPK signaling pathway", etc. The RT-qPCR results showed that the expressions of hsa_circRNA_001544 (U = 486.5, P < 0.05) and hsa_circRNA_102532 (U = 645, P < 0.05) were significantly different between the AS group and the HC group. The AS group was further divided into two subgroups: active AS (ASA) and stable AS (ASS). After analysis, it was found that compared with the HC group, hsa_circRNA_001544 was significantly increased in both ASA (U = 214, P < 0.05) and ASS groups (U = 273, P < 0.05), while hsa_circRNA_008961 (U = 250, P < 0.05) and hsa_circRNA_102532 (U = 295, P < 0.05) were only significantly increased in the ASA group. Furthermore, hsa_circRNA_012732 was significantly different between the ASA and ASS groups (U = 194, P < 0.05), and there was no statistical significance among the remaining groups. Correlation analysis results showed that hsa_circRNA_012732 was negatively correlated with Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), high-sensitivity C-reactive protein (hsCRP), and globulin (GLOB) and positively correlated with lymphocyte count (LY), mean corpusular volume, and albumin (ALB), and hsa_circRNA_008961 was negatively correlated with platelet (PLT) count. ROC curve analysis showed that hsa_circRNA_001544 (95% CI = 0.610-0.831, P < 0.05) and hsa_circRNA_102532 (95% CI = 0.521-0.762, P < 0.05) were statistically significant, and their area under curve (AUC) values were 0.720 and 0.642, respectively. CONCLUSIONS There are differentially expressed circRNAs in PBMCs of AS patients, and they may be involved in the occurrence and development of AS. Among these differentially expressed circRNAs, hsa_circRNA_012732 has the potential to become an indicator of disease activity, and hsa_circRNA_001544 has the potential to become a molecular marker for AS diagnosis.
Humans ; Leukocytes, Mononuclear ; RNA/genetics* ; RNA, Circular ; ROC Curve ; Spondylitis, Ankylosing/genetics*

OBJECTIVETo investigate the association between heat shock protein 70-hom (HSP70-hom) gene polymorphism and ankylosing spondylitis(AS) in Chinese Han patients.

METHODSGenomic DNA from 98 Chinese AS patients and 70 ethnically matched controls were typed for HSP70-hom polymorphism by polymerase chain reaction-restriction fragment length polymorphism.

RESULTSThe HSP70-hom genotypes in the AS patients consisted of homozygote AA (60.2%) and BB(4.1%), and heterozygote(35.7%), while the HSP70-hom genotypes in the controls were composed of AA(58.6%), BB(2.9%) and heterozygote(38.6%). No significant difference was found in the distribution of HSP70-hom genotype between these two groups(chi(2) test=0.280, P>0.05). The frequencies of HSP70-hom alleles in AS patients were 77.9%(AA) and 22.1%(BB), while they were 78.1% and 21.9% in the controls. The frequency of HSP70-hom allele in AS patients was not significantly increased, compared with that in controls (chi(2) test=0.002, P>0.05).

CONCLUSIONThere may be no association between the HSP70-hom gene polymorphism and ankylosing spondylitis in Chinese Han population.

Adolescent ; Adult ; Aged ; Child ; Female ; HSP70 Heat-Shock Proteins ; genetics ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Spondylitis, Ankylosing ; genetics

OBJECTIVE: To explore the correlation of polymorphisms of AF4/FMR2 family genes and IL-10 gene with genetic susceptibility to ankylosing spondylitis (AS) and identify the high-risk factors of AS. METHODS: This case-control study was conducted among 207 AS patients and 321 healthy individuals. The tag single nucleotide polymorphisms (SNPs) rs340630, rs241084, rs10865035, rs1698105, and rs1800896 of the AF4/FMR2 family gene and IL-10 gene of the AS patients were genotyped, and the distribution frequencies of the genotypes and alleles were analyzed to explore the relationship between different genetic models and AS and the gene-gene and gene-environment interactions. RESULTS: Gender ratio, smoking history, drinking history, hypertension, erythrocyte sedimentation rate and C-reactive protein differed significantly between the case group and the control group (P < 0.05). The dominant model and recessive model of AFF1 rs340630, the recessive model of AFF3 rs10865035, and the recessive model of IL-10 rs1800896 were significantly different between the two groups (P=0.031, 0.010, 0.031, and 0.019, respectively). Gene-environment interaction analysis suggested that the interaction model incorporating AFF1 rs340630, AFF2 rs241084, AFF3 rs10865035, AFF4 rs1698105, IL-10 rs1800896, smoking history and drinking history was the best model. The genes related with AF4/FMR2 and IL-10 were enriched in the biological processes of AF4 super extension complex, interleukin family signal transduction, cytokine stimulation and apoptosis. The expression levels of AF4/FMR2 and IL-10 were positively correlated with immune infiltration (r > 0). CONCLUSION The SNPs of AF4/FMR2 and IL-10 genes are associated with the susceptibility to AS, and the interactions of AF4/FMR2 and IL-10 genes with the environmental factors contributes causes AS through immune infiltration.
Humans ; Case-Control Studies ; Genetic Predisposition to Disease ; Interleukin-10/genetics* ; Polymorphism, Single Nucleotide ; Spondylitis, Ankylosing/genetics* ; Transcriptional Elongation Factors/genetics* ; Nuclear Proteins/genetics*

OBJECTIVETo investigate the frequencies distribution of human leukocyte antigen (HLA)-B27 subtypes in unrelated healthy Chinese.

METHODPolymerase chain reaction sequence-based typing (PCRSBT) was used to determine HLA high-resolution genotypes of 825 unrelated healthy Chinese.

RESULTSA total of 25 HLA-B27-positive individuals and 8 HLA-B27 subtypes were detected. These subtypes and their corresponding frequencies were B * 2704 (30.77%) , B * 2705 (23.08%), B * 2707 (19.23%), B * 2711 (7.69%), B * 2712 (7.69%), B * 2701 (3.85%), B * 2713 (3.85%) and B * 2721 (3.85%).

CONCLUSIONThe data obtained through PCR-SBT method may serve as important reference for the research of relationship between HLA-B27 subtypes and some diseases such as ankylosing spondylitis.

Adult ; Asian Continental Ancestry Group ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; HLA-B27 Antigen ; classification ; genetics ; Humans ; Male ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Spondylitis, Ankylosing ; genetics

BACKGROUNDAnkylosing spondylitis (AS) is the most common rheumatic condition that is slowly progressive and predominantly affects adolescents. Pathological bone formation associated with AS is an important cause of disability. The aim of the study was to investigate the possible involvement of the genes related to endochondral ossification and ectopia ossification in genetic susceptibility to AS in a Chinese Han population.

METHODSSixty-eight single nucleotide polymorphisms (SNPs) from 13 genes were genotyped in discovery cohorts including 300 AS patients and 180 healthy controls. The rs10019009 in dentin matrix protein 1 (DMP1) gene shown as association with AS after multiple testing corrections in discovery cohorts was replicated in a validation independent cohort of 620 AS patients and 683 healthy controls. The rs10019009 was assessed with bioinformatics including phylogenetic context, F-SNP and FastSNP functional predictions, secondary structure prediction, and molecular modeling. We performed a functional analysis of rs10019009 via reverse transcription-polymerase chain reaction, alkaline phosphatase (ALP) activity in human osteosarcoma U 2 OS cells.

RESULTSInterestingly, the SNP rs10019009 was associated with AS in both the discovery cohort (P = 0.0012) and validation cohort (P = 0.0349), as well as overall (P = 0.0004) in genetic case-control association analysis. After a multivariate logistic regression analysis, the effect of this genetic variant was observed to be independent of linkage disequilibrium. Via bioinformatics analysis, it was found that the amino acid change of the rs10019009 led to changes of SNP function, secondary structure, tertiary conformation, and splice mode. Finally, functional analysis of rs10019009 in U 2 OS cells demonstrated that the risk T allele of the rs10019009 increased enzymatic activity of ALP, compared to that of the nonrisk allele (P = 0.0080).

CONCLUSIONSThese results suggested that the DMP1 gene seems to be involved in genetic predisposition to AS, which may contribute to the ectopic mineralization or ossification in AS. In addition, DMP1 gene may be a promising intervention target for AS in the future.

Adult ; China ; ethnology ; Extracellular Matrix Proteins ; chemistry ; genetics ; Female ; Genetic Predisposition to Disease ; Humans ; Logistic Models ; Male ; Phosphoproteins ; chemistry ; genetics ; Polymorphism, Single Nucleotide ; Spondylitis, Ankylosing ; etiology ; genetics

OBJECTIVETo investigate the pathological expression and significance of VEGF in patients with active ankylosing spondylitis.

METHODSThe expression of VEGF in the synovial tissues of cacroiliac joint of patients with active AS was detected by using in situ hybridization and the results were compared with those in the patients with pelvic fracture using image analysis system.

RESULTSThe positive expressions of VEGF in the synovial tissues of cacroiliac joint of patients with active AS were stronger than those in the control group (P<0.01).

CONCLUSIONVEGF are important factors in patients with active AS. They are tightly correlated with the process of osteoclasia and pathological new bone formation in the cacroiliac joint of patients with active AS. If we can reduce the expressions of VEGF in the patients with active AS, the process of osteoclasia and pathological new bone formation will be interrupted and this provides a new strategy for the treatment of ankylosing spondylitis.

Adult ; Case-Control Studies ; Female ; Gene Expression Regulation ; Humans ; Male ; RNA, Messenger ; genetics ; metabolism ; Spondylitis, Ankylosing ; genetics ; Synovial Fluid ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVETo confirm the role of HLA-B2704 and hbeta(2)m gene in the pathogenesis of spontaneous inflammatory diseases by establishing HLA-B2704 and hbeta(2)m double transgenic mice model of ankylosing spondylitis. It will provide a powerful animal model for exploring the etiology, prevention and treatment of B27-relevant diseases.

METHODSThe screening, identification and expression of HLA-B2704 and hbeta(2)m gene were determined by PCR, dot blot, Southern blot hybridization, RT-PCR, flow cytometry and immunohistochemistry. HE staining was performed for the diseased mice.

RESULTSEight double transgenic mice bearing high copy developed spontaneous dermatosis, arthritis and nail changes in the rear paw. The results of flow cytometry in normal mice, B27 single transgenic mice, and HLA-B27/hbeta(2)m double transgenic mice were 0.63%, 7.87% and 35.87% respectively. HLA-B2704 antigen was high expressed on the cell surface, but not evident on those of B27 single transgenic mice.

CONCLUSIONSHLA-B2704 heavy chain can induce spontaneous inflammatory diseases in the transgenic mice. Hbeta(2)m can form a stable complex with HLA-B27 and may stabilize and enhance the expression of HLA-B2704 on the cell surface.

Animals ; Disease Models, Animal ; HLA-B27 Antigen ; genetics ; Inflammation ; etiology ; Mice ; Mice, Transgenic ; Reverse Transcriptase Polymerase Chain Reaction ; Spondylitis, Ankylosing ; genetics ; physiopathology

BACKGROUND: HLA-B27 is strongly associated with ankylosing spondylitis (AS), and its subtypes differ in their ethnic distribution. Studies worldwide have shown that B*2701, B*2702, B*2704, B*2705, B*2707, B*2708, B*2714, B*2715, and B*2719 are AS-predisposing subtypes, whereas B*2706 and B*2709 are reported to be negatively associated with AS. The aim of this study was to investigate HLA-B27 polymorphism and clinical features according to subtypes in Korean patients with AS. METHODS: Two hundred thirty samples from patients with impression of AS were analyzed by polymerase chain reaction using a sequence-specific primers (PCR-SSP) method. Pel-Freez SSP Unitray HLA-B*27 kit (Dynal Biotech, USA) including 16 primers was used to define HLA-B27 subtypes from B*2701 to B*2735. RESULTS: Among 230 samples from patients with impression of AS, 171 were HLA-B27 positive, and among 160 patients diagnosed as AS, 154 (96.3%) were HLA-B27 positive, while 17 patients not diagnosed as AS were HLA-B27 positive. Among 154 HLA-B27 positive patients with AS, 142 (92.2%) were typed as B*2705 and 9 (5.8%) were typed as B*2704. Three cases (1.9%) could be interpreted only variously because of their HLA-B27 homogeneous alleles. Between B*2705 and B*2704, no specific HLA-B27 subtype appeared to contribute to AS susceptibility (P=0.60). Difference in clinical features between B*2705 and B*2704 could not be found in this study (P>0.05). CONCLUSIONS: This study verified that HLA-B27 (96.3%) is strongly associated with AS and identified that the major subtypes of HLA-B27 positive patients with AS in Korea are B*2705 (92.2%) and B*2704 (5.8%).
Adult ; Alleles ; Female ; Gene Frequency ; Genotype ; HLA-B27 Antigen/blood/*genetics ; Humans ; Korea/epidemiology ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Reagent Kits, Diagnostic ; Spondylitis, Ankylosing/*diagnosis/epidemiology