Objective To consecutively understand the national clinical testing status and to reassure the quality-control of autoantibody detection. Methods Letter or telephone notification were conducted to the participating hospitals or departments. Autoantibodies for quality control survey included anti-nuclear anti-body (ANA), anti-double-stranded DNA (anti-dsDNA), anti-extractable nuclear antigens (A-ENA), anti-mitochondria antibody (AMA)/anti-smooth muscle antibody (ASMA), and anti-citrulline antibody (anti-CCP). Each had 3 control samples, and altogether 15 samples for testing. Sample distribution and data analysis were double-blinded. Results One hundred and two hospitals/departments participated in the national quality-control survey. The accurate rate for this survey was 70%, 88%, 93%, 85%, 79% respectively for ANA, anti-dsDNA, AMA, ASMA and anti-CCP. Anti-ENAs were further divided into anti-RNP, Sm, SSA, SSB and Scl-70 subgroups, and the accurate rate was 88%, 77%, 92%, 93% and 87% respectively. Conclu-sion Compared to the previous 4 national surveys, the accurate rates of ANA, anti-dsDNA, anti-ENAs, AMA in our country's autoantibody testing is improved, and the detection rate of ASMA and anti-CCP antibody is also increased. More hospitals and testing items can test these autoantibodies, which implies that autoantibody testing status has been improving in our country.

Objective To investigate the changes of clinical and laboratory manifestations of rheumatic fever (RF) in recent ten years by reviewing the medical data of 315 patients with RF. Methods Three hundred and fifteen in-patients and out-patients with RF during 1985-1995 (group A) and 1997-2007(group B) were selected. Their manifestations were compared. Results Male/female ratio was about 1:2. Carditis and polyarthritis were common manifestations. Compared with group A, the rate of low-grade fever and carditis increased and the rate of heart failure, positive rate of C reaction protein and antistreptolysin O decreased in group B. In group B, 61.4% patients fulfilled the updated Jones diagnostic criteria. 76.2% fulfilled the 2002-2003 WHO criteria. The sensitivity and specificity of peripheral blood lymphocyte procoagulant activity (PCA) for the diagnosis of rheumatic carditis was 79.1% and 71.4% respectively. That of the anti-streptococcal group A polysaccharide (ASP) antibodies was 70.3% and 70% respectively. Five to ten years follow-up clinical data were available for 35 cases since Dec. 1997. The recurrent rate of RF was 62.8%. Only 1/3 cases received regular secondary prevention. Recurrence rate of patients with regular secondary prevention was significantly lower than that of patients without regular secondary prevention. Conclusion Mild earditis has been increasing during last ten years. PCA and ASP are valuable tests for diagnosing rheumatic carditis. More emphasis should be paid to atypical cases, early diagnosis and regular secondary prevention in order to improve prognosis.

Objective To investigate the mechanism of brain derived neurotrophic factor (BDNF) regulated by differ-ent isoforms of tyrosine kinase receptor B (TrkB) in epileptic hippocampal neurons. Methods Primary hippocampal neu-rons were cultured in vitro for 7 days, and divided into two groups, ALLN (calcineurin inhibitor) group and Anisomycin (trans-lation inhibitor) group. ALLN group included control group, control+BDNF group, epilepsy group, epilepsy+BDNF group, control+ALLN group, epilepsy+ALLN group and epilepsy+ALLN+BDNF group. Anisomycin group was sub-divided into con-trol group, control+BDNF group, epilepsy group, epilepsy+BDNF group, control+Anisomycin group, epilepsy+Anisomycin group and epilepsy+Anisomycin+BDNF group. The immunofluorescent technique was used to identificate the hippocampal neurons. Epileptiform discharges were detected by electrophysiological techniques. Western blot assay was used to deter-mine the protein expression of TrkB and phosphorylated TrkB (p-TrkB) in all cell groups. Results (1) In ALLN group, the gray value of p-TrkB/TrkB was higher in control+BDNF group compared with that of control group, the value was higher in epilepsy+BDNF group than that of epilepsy group but was lower than that of control+BDNF group. The gray value of p-TrkB/TrkB was lower in epilepsy+ALLN+BDNF group than that of epilepsy+BDNF group, but no significant difference compared with that of epilepsy+ALLN group. (2) In Anisomycin group:the gray value of p-TrkB/TrkB was higher in control+BDNF group than that of control group. The gray value of p-TrkB/TrkB was higher in epilepsy+BDNF group than that of epilepsy group, but which was lower than that of control+BDNF group. The gray value of p-TrkB/TrkB was higher in epilepsy+Aniso-mycin+BDNF group than that of epilepsy+BDNF group and epilepsy+Anisomycin group. Conclusion The decreased ex-pression of TrkB.T can improve the inhibition of BDNF/TrkB signaling, and BDNF can activate BDNF/TrkB signal pathway in epileptic hippocampal neurons. The increased TrkB.FL protein level by ALLN can’t improve the inhibition of BDNF/TrkB signal pathway.

Objective To study the effect of miR-204 on BDNF/TrkB signaling and pathogenesis on the neuron model of epilepsy. Methods Primary hippocampal neurons were cultured in vitro for 7 days, and were divided into control group, control+BDNF group, epilepsy group, epilepsy+BDNF group , control+miR204 group, epilepsy+miR204 group and ep-ilepsy+miR204+BDNF group. The epilepsy model of hippocampal neurons was established by being exposed to Mg2+free me-dia for 3 hours. The miR-204 lentivirus vector was constructed. The effect of miR-204 on BDNF/TrkB expression was detect-ed by immunohistochemistry, patch clamp and Western blot technique. Results Compared with the control group, the TrkB phosphorylation level was higher in control+BDNF group. The TrkB phosphorylation level was lower in epilepsy+BDNF group than that of control+BDNF group, but it was higher than that of epilepsy group. The TrkB phosphorylation level was higher in epilepsy+miR204+BDNF group than that of epilepsy+BDNF group and epilepsy+ miR204 group. Conclusion BDNF and miR-204 can improve the inhibitory condition of BDNF/TrkB signaling and may play an important role in alleviat-ing epilepsy disease.

BACKGROUND: Human leukocyte antigen (HLA)-B27 is closely connected to the occurrence of some rheumatic diseases such as ankylosing spondylitis and can be used as an important factor for evaluating the diagnosis of ankylosing spondylitis. Immunomagnetic separation and enzymelinked immunosorbent assay (IMS-ELISA) has been applied to the detection of HLA-B27.OBJECTIVE: To explore the accuracy, sensitivity and specificity of IMSELISA for detecting HLA-B27 and its value in the auxiliary diagnosis of ankylosing spondylitis.DESIGN: A clinical trial in comparison with the gold standard.SETTING: Departments of Rheumatology and Clinical Laboratory, Third Affiliated Hospital of Sun Yat-sen University.PARTICIPANTS: Eighty-six patients suffering from low back pain and/or arthritis who were treated for the first time in Department of Rheumatology from December 2002 to April 2003. Inclusion criteria: ① Presence of manifestations of low back pain and/or arthritis; ② Thorough documentation of clinical and other examinations; ③ Informed consent to HLA-B27 examination; ④ Treatment for the first time in the Third Affiliated Hospital of Sun Yet-sen University. Those with other serious diseases or with incomplete record of clinical and/or accessory examinations were excluded. The 86 patients included 56 male and 30 female patients aged from 12 to 65 years.METHODS: Blood sample was detected for HLA-B27 by both IMS-ELISA and microlymphocytotoxicity test, and the latter was selected as the gold standard. The coincidence rate of the results detected by the two methods as well as the sensitivity, specificity, positive and negative predictive values of IMS-ELISA were calculated.MAIN OUTCOME MEASURES: ① The coincidence rate of the results of the two methods. ② The sensitivity and specificity of IMS-ELISA for detecting HLA-B27.RESULTS: None of the patients was lost. For the 33 patients with ankylosing spondylitis, the positivity rate of IMS-ELISA (90.9％) was higher than that of microlymphocytotoxicity test (87.9％), but the difference was not statistically significant (P＞0.05). The total coincidence rate of the two methods was 93.0％ in all the 86 patients. The sensitivity, specificity, positive and negative predictive values of IMS-ELISA were 90.0％, 95.7％,94.7％ and 91.7％ respectively.CONCLUSION: Both IMS-ELISA and microlymphocytotoxicity test are capable of reliable examination of HLA-B27 with high sensitivity and specificity.

Objective To observe the effect of infliximab combination therapy on the expression of CD147 on the peripheral CD14+ monocytes of active rheumatoid arthritis (RA) patients. Methods Thirty active RA patients who were refractory to MTX treatment were randomized into three groups (group A, B, C) with the proportion of 3:1:1. Group A and B received four or three infusions of infliximab (3 mg/kg), group C received four infusions of placebo. All three groups were added to a stable background of MTX. The mean fluorescence intensity (MFI) of CD147 expression on the peripheral CD14+ monocytes of RA patients and normal healthy controls were detected by flow cytometry analysis. Clinical and laboratory parameters were assessed before each infusion. One-way ANOVA, Kruskal-Wallis the MFI of CD147 expression at week 18 (P<0.05). Marked differences were observed between the infliximab + MTX group and the placebo + MTX group on the change of the MFI of CD147 expression from baseline to week 18 (P<0.05).Conclusion CD147 expression on the peripheral CD14+ monocytes of active RA patients is increased, and combination therapy of infliximab and MTX can inhibit the expression.

Objective To study the single nucleotide polymorphisms (SNPs) in IL-23R gene in Chinese Han population with ankylosing spondylitis (AS). Methods SNPs rs11209026, rs1343151, rs11209032 and another three SNPs near them based on their physical distances were genotyped by PCR-directed sequencing. Hardy-Weinberg equilibrium, genotypes and allele frequency analysis were analyzed by SPSS 13.0. Linkage disequilibrium and haplotype analysis were carried out by SHEsis software. Results The difference of genotypes of rs11209032 and the difference of genotypes and allele frequencies of rs6677188 between patients and controls were statistically significant (P＜O.01) ; The two SNPs rs11209032 and rs6677188 had strong linkage disequlibfium (D'=0.925, r2=0.561 ). Haplotype analysis had shown a higher proportion of GAC haplotype in patients and a higher proportion of GTC haplotype in controls. Conclusion These results suggest that IL-23R polymorphisms is associated with susceptibility to AS in Chinese Han population and IL-23R gene may be a susceptible gene of AS.

Objective To further investigate the effect of sinomenine (SIN) on TNF-α-induced VCAM-1 expression in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were isolated from freshly collected umbilical cords. Positive control samples were stimulated with TNF-α, but free of SIN. Negative control samples were treated in the same way, but without TNF-α and SIN. Experimental samples were co-cultured with TNF-α and SIN at various concentrations (0.25, 0.5, and 1.0 mol/L), or TNF-α and dexamethasone (Dex) at concentration of 1.0×10-6 mol/L, or TNF-α with Dex (at concentration of 1.0×10-6mol/L) and SIN at different concentrations (0,25, 0.5, and 1.0 mmol/L) (co-treated groups). VCAM-1 expression was detected by flow cytometry (FCM). Results SIN inhibited expression of VCAM-1 in TNF-α-induced HUVECs, the best effect was shown in the 1.0 mmol/L SIN treated group. VCAM-1 decreased more markedly in the co-treated groups. Conclusion SIN inhibits TNF-α-induced VCAM-1 expression on HUVECs in vitro, and SIN maybe synergistic with Dex in inhibiting TNF-α-induced VCAM-1 expression on HUVECs in vitro.

Objective To investigate the reasons for delayed diagnosis of ankylosing spondylitis(AS) in a Chinese population.Methods Three hundred and eight patients fulfilled the 1984 Modified New York criteria of AS were enrolled.They were interviewed in person or by telephone by rheumatologists for 13 ques-tions.Results Of the 308 AS patients.238(75%)completed 13 questions.Among these 238 patients,male to female ratio was 6 to 1.The average age at AS onset was 22.1 years.Those aged under 15 years at disease onset Was 18.1%.and between 15 and 39 years was 79.0%while over 39 years was 2.9%.Of these 238 pa-tients.27(23.9%)had family history of AS.84.9% of the patients were HLA-B27 positive.The average dura-tion of delayed diagnosis in HLA-B27(+)and HLA-B27(-)were 70.1 and 88.1 months respectively,which was not significant statistically.Among those delayed over 10 vears,24 AS patients were HLA-B27(+)and 10 were HLA-B27(-),which was statistically significant(P=0.012).66.4% of 238 patients were misdiagnosed, of which 23.4%were diagnosed as arthritis associated with rheumatic fever,22.9% as fatigue and 20.3%as inter-vertebrate disc.45.0%were misdiagnosed by one physician while 2 1.4%were misdiagnosed for several times,and the average length of delay diagnosis was 72±68 months.Seven cases,although the diagnosisi was being delaved for more than 10 years the initial diagnosis was correct. Conclusion Delayed diagnosis of AS is common in China.The major reasons for delayed diagnosis are HLA-B27 negative and lack of X-ray changes of sacroiliac joint at the initial visit.

Objective To investigate the distribution of HLA-B27 subtypes in ankylosing spondylitis(AS) patients of Chinese Han population by using the updated HLA-B27 typing data. Methods One hundred AS subjects were randomly selected from spondyloarthritis patients data bank of the third affiliated hospital of Sun Yat-sen university. All subjects were independent individuals, and the duplicated samples in the same family were excluded. Salt fraction method was used to prepare genome DNA. Luminex liquid array combining PCR-SSOP was used to perform the low resolution HLA-B genotyping. PCR-SSP was applied to perform the high resolution HLA-B27 typing for HLA-B27 positive subjects. Results Ninety-eight independent AS patients were recruited randomily, of which, 93 were HLA-B27 positive, with positive rate 94.9%, and covered 96% patients with family history of AS. Three subtypes were detected in this population including B * 2704 (n=76, 81.7%), B * 2705 (n=12, 12.9%) and B * 2715 (n=5, 5.4%). Compared with the two reports about HLA-B27 subtype distribution in healthy HLA-B27 positive Han population there was no significant difference between AS patients and healthy controls. But no B * 2715 case was found in those two reports of healthy population. Three reports (including 1 report in Chinese) could found about B * 2715 subtype, but all positive cases were oriental people. Furthermore, all B * 2715 positive patients were AS patients. Conclusion B * 2704 is the predominant subtype ,in AS patients of Chinese Han population, and followed by B * 2705. We found five cases with positive B * 2715, a considerable rare allele. This may suggest association between B * 2715 and AS.