Objective To evaluate the distribution and significance of IgG subclasses of anti-cyclic cirullinated peptide antibody (anti-CCP) in sera from patients with rheumatoid arthritis (RA).Methods A total of 83 patients with RA at the Department of Endocrinology of Taizhou Hospital , 51 disease controls and 50 healthy controls during the period from August 2012 to June 2013 were enrolled.The total serum IgG and IgG subclasses of anti-CCP antibodies were detected by antigen specific enzyme linked immune-sorbent assay( ELISA ).The prevalence and relative amount of IgG subclasses were calculated and compared.Statistical analysis was performed by χ2 test and Kruskal-Wallis H test.Results The positive rates of IgG subclasses of anti-CCP were anti-IgG 71.1%(59/83), anti-IgG1 78.3%(65/83), anti-IgG2 26.5%(22/83), anti-IgG3 60.2%(50/83), anti-IgG4 74.7%(62/83) respectively.The diagnostic value of anti-CCP-IgG1, anti-CCP-IgG3 and anti-CCP-IgG4 alone or combined (AUC =0.818-0.901),compared with anti-CCP-IgG(AUC=0.857), had no significant difference(Z=0.028-0.045,P>0.05).The DAS28 score of anti-CCP-IgG1(DAS28 =6.5), and anti-CCP-IgG4(DAS28 =6.5)positive in patients with RA were significantly higher than those in negative groups (DAS28=4.5,4.6)(U=396.0,427.5,P<0.01).The T28(T28=4.0,4.0)and SW28(SW28=4.0,4.0) results of CCP-IgG1and CCP-IgG4 positive in patients with RA were significantly higher than those in negative groups (T28=3.0,3.0,SW28 =3.0,3.0)(U=377.5,406.0,255.5,286.5,P<0.05).Conclusions The distribution of IgG subclasses of anti-CCP in sera from patients with RA was predominantly anti-CCP-IgG1, anti-CCP-IgG3 and anti-CCP-IgG4 associated with RA disease activity.However , whether joint detection of IgG subclasses can replace conventional anti -CCP is questionable.

Objective Marginal division (MrD) of striatum is a universal structure in the mammalian brain,and it plays an critical role in learning and memory.In the present study,we try to investigate the synaptic ultrastructure of Methionine enkephalin (MET-ENK) immunoreacted fibers connected with neurons in the marginal division of the striatum in the monkey brains to explore the ultrastructural basis of the mechanism of learning and memory function in MrD.Methods Six male monkeys (macaque) were perfused with paraformaldehyde through heart to fix the brain and the brains were sectioned by a cryostat.Sections of the brains were performed immunohistochemical staining to detect the MET-ENK expression in the stfiatum; the areas with positive immumohistochemical staining was performed ultrastructural observation for morphological characteristics of the MET-ENK synapses in the MrD.Results Immunohistochemistry staining showed a dense arrangement of MET-ENK immunoreactive cells between the putamen and globus pallidus.Five major types of MET-ENK synapses were identified in the MrD:the axo-dendritic synapses,the axo-spinous synapses,the axo-somatic synapses,the axo-axonic synapses and the compound synapses.Conclusion The MET-ENK synapses in the MrD are diverse and complex,and can be distinguished from the rest of the striatum.

OBJECTIVEInterleukin-27 (IL-27) has been reported to reduce the levels of interleukin-17 (IL-17) and alleviate the severity of experimental autoimmune myocarditis. IL-17, an important tissue-protective cytokine in viral myocarditis (VMC), has been reported to increase synovial expression of IL-27 in rheumatoid arthritis. However, the influence of IL-17 on IL-27 expression in murine model of VMC remains unknown.

METHODSWild-type (WT) and IL-17A-deficient (IL-17A(-/-)) mice on the BALB/c background were intraperitoneally (i.p) injected with coxsackievirus B3 (CVB3) for establishing VMC models. Cardiac tissue was obtained on day 7 after CVB3 injection. Myocardial histopathologic changes were observed by hematoxylin-eosin (HE) stained myocardial sections.Expression of IL-27 in heart and serum was measured by real-time reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA), respectively. Furthermore, splenic lymphocytes and peritoneal macrophages were purified 1 week after injection from WT mice.Isolated lymphocytes were cultured in the presence of different concentrations (0 and 25 ng/ml) of recombinant IL-17 (rIL-17) for 24 h. Macrophages were cultured with different concentrations of rIL-17 (0 and 10 ng/ml) for 48 h.IL-27 mRNA expression of cultured cells was assayed by RT-PCR, and their protein level in the culture supernatant was measured by ELISA.

RESULTSCompared with WT mice, significantly less cardiac inflammation was evidenced in the heart of IL-17A-/- mice (0.9 ± 0.3 vs.1.9 ± 0.5) , relative cardiac IL-27 p28 mRNA expressions (1.11 ± 0.24 vs.3.1 ± 0.8) and serum IL-27 protein[(72 ± 18) pg/ml vs.(95 ± 25) pg/ml] were also significantly lower in IL-17A-/- mice (all P < 0.05).In the culture lymphocytes, the relative mRNA (1.02 ± 0.13 vs.1.32 ± 0.21) and protein [(49 ± 9) pg/ml vs.(52 ± 11) pg/ml]expressions of IL-27 p28 and were similar post treatment with 0 and 25 ng/ml rIL-17 (all P > 0.05). Compared with 0 ng/ml rIL-17 culture with macrophages, higher relative mRNA (8.5 ± 3.1 vs.2.2 ± 0.7) and protein [(368 ± 95) pg/ml vs.(150 ± 38) pg/ml] expressions of IL-27 p28 were detected in 10 ng/ml rIL-17 group (all P < 0.05).

CONCLUSIONOur data indicates that cytokine IL-17 may contribute to the secretion of IL-27 in VMC mice.Furthermore, macrophages but not lymphocytes may be the important IL-27-producing immune cells and major target cells for IL-17. Thus,IL-27 and IL-17 might be actively involved in the pathogenesis of VMC.

Animals ; Coxsackievirus Infections ; immunology ; metabolism ; Disease Models, Animal ; Interleukin-17 ; immunology ; Interleukin-27 ; metabolism ; Macrophages ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis ; immunology ; metabolism