Objective To study the expression and value of CXCR1 and CXCR2 on neutrophils, CD14~+ monocytes and CD3~+ T lymphocytes of peripberol blood of ankylosing spondylitis(AS)patients and to investigate the correlation between CXCR1,CXCR2 and disease activity.Methods A case control study was designed and enrolled 30 active AS,30 active rheumatoid arthritis(RA)and 30 healthy controls.The levels of CXCR1 and CXCR2 expression on neutrophils,CD3~+ T cells and CD14~+ monocytes of peripheral blood of the patients and healthy controls enrolled were measured and analyzed by flow cytometry by measuring the mean fluorescence intensity(MFI)channel.The correlations between the level of CXCR1 and CXCR2 anti disease activity or functional index of AS such as BASDAI,BASF1,ESR and CRP were analyzed.Results The MFI of CXCR1 expression on CD3~+ T lymphocytes of peripheral blood was significantly higher in AS patients (41?24)than that in RA patients(18?10)and healthy controls(19?7)(P

Objective To conduct mutation screening of SCN1A 3′ untranslated region (UTR) on Dravet syndrome (DS) patients without mutations in the SCN1A coding region and promoter region, and functional analysis of the variant from DS patients.Methods Twenty-eight DS patients without mutations in the SCN1A coding region and promoter region were screened for SCN1A 3′ UTR mutations using PCR and direct sequencing.Functional analysis of the detected mutation was done via luciferase assay, mRNA stability analysis and RNA electrophoretic mobility shift assay (RNA-EMSA).Results A novo variant (c.*20A>G) in SCN1A 3′ UTR was found in one DS patient.The variant (c.*20A>G) reduced the luciferase gene xpression by 30% through increasing the affinity of pluripotent embryonal carcinoma cell line NT2/cytoplasmic protein binding and reducing luciferase gene mRNA stability (t=8.5,P<0.01).Conclusions A functional variant was detected from one patient with DS.This variant negatively regulated the gene expression by increasing the affinity of pluripotent embryonal carcinoma cell line NT2/cytoplasmic protein binding and reducing mRNA stability.

OBJECTIVETo screen the parameters of questionnaire in prevalence survey of spondyloarthropathy (SpA) in China, and to evaluate the value and feasibility of questionnaire used in prevalence survey.

METHODSA questionnaire study on SpA with 12 questions involved was performed which came from a epidemiological survey on SpA in Brittany, France.

RESULTS(1) We found difference on the sensitivity and specificity of some indexes in the questionnaires between the French study and the one developed by ourselves. The sensitivity differed between the published French paper and ours in the following indexes: onset age, psoriasis and inflammatory bowel disease. The specificity of the indexes would include spinal pain, insidious onset, morning stiffness, duration more than 3 months and radiographic manifestation also showed differences. (2) Excluding the radiographic abnormality, we ran the logistic regression and concluded that the following parameters were the independent indexes which suggesting the existence of the disease: spinal pain onset before 40 years of age, having spinal stiffness in the morning having positive family history and having buttock pain and heel pain. (3) Based on the result of each question of the questionnaire, indices of distinguishing the cases and controls were identified.

CONCLUSIONThe questionnaire verified in our study was a new, simple, valuable and feasible one for SpA prevalence study and in screening the potential SpA patients.

Adult ; China ; epidemiology ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Spondylarthropathies ; epidemiology ; Surveys and Questionnaires

BACKGROUNDRheumatoid arthritis (RA) is an autoimmune disease characterized by chronic inflammation at the synovial membrane. Although great progress has been made recently in exploring the etiology and pathogenesis of RA, its molecular pathological mechanism remains to be further defined and it is still a great challenge in determining the diagnosis and in choosing the appropriate therapy in early patients. This study was performed to screen candidate RA-associated serum proteins by comparative proteomics to provide research clues to early diagnosis and treatment of RA.

METHODSSera isolated from 6 RA patients and 6 healthy volunteers were pooled respectively and high-abundance proteins were depleted by Plasma 7 Multiple Affinity Removal System. The protein expression profiles between the two groups were then compared by two-dimensional gel electrophoresis (2-DE) and the proteins over/under-expressed by more than 3-fold were identified by mass spectrometry analysis. To validate the differential expression levels of the identified proteins between the two groups, ELISA was performed in two of the identified proteins in individual sera from 32 RA patients and 32 volunteers.

RESULTSEight proteins which over/under-expressed in sera of RA patients were identified. Among them, chain A of transthyretin (TTR) was under-expressed, while serum amyloid A protein, apolipoprotein A (ApoA)-IV, ApoA-IV precursor, haptoglobin 2, ceruloplasmin (Cp), immunoglobulin superfamily 22 and HT016 were over-expressed. ELISA test confirmed that Cp expressed remarkably higher while TTR obviously lower in RA group compared with volunteer group.

CONCLUSIONThere were 8 identified proteins differentially expressed between RA group and volunteer group, which might be candidate RA-associated proteins and might be promising diagnostic indicators or therapeutic targets for RA.

Adult ; Apolipoproteins A ; blood ; Arthritis, Rheumatoid ; blood ; Blood Proteins ; analysis ; Ceruloplasmin ; analysis ; Electrophoresis, Gel, Two-Dimensional ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Prealbumin ; analysis ; Proteomics ; Serum Amyloid A Protein ; analysis

BACKGROUNDThe role of B-cell remains an enigma in the pathogenesis of ankylosing spondylitis (AS). This study aimed to investigate the distributions of B-cells and subsets in peripheral blood of AS patients and observe their changes in etanercept-treated AS patents.

METHODSWe detected the proportions of CD19(+) B-cell, naive B-cell (CD19(+)CD27-), memory B-cell (CD19(+)CD27dim) and plasmablast (CD19(+)CD27high) in peripheral blood of 66 patients with AS (39 at active stage, 27 at stable stage; 35 patients with peripheral joint involvement, 31 patients with axial involvement alone), 30 patients with rheumatoid arthritis (RA) and 30 healthy volunteers using flow cytometry. And then we observed the changes of the above indexes of 39 active AS patients treated with etanercept in a randomized, double-blind, placebo-controlled trial.

RESULTS(1) Percentages of CD19(+) B-cells in active or peripheral joint involvement AS patients increased more obviously than those in stable or axial involvement alone AS patients (both P = 0.001), and percentage of CD19(+)CD27high B-cells in AS patients with peripheral joint involvement was significantly higher than that in cases with axial involvement alone or healthy volunteers (P = 0.005 and 0.006, respectively); (2) The percentage of CD19(+) B-cells in AS patients was positively correlated with Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) scores, Patient's Global Assessment (PGA) scores, total back pain scores and nocturnal back pain scores (r = 0.270, 0.255, 0.251 and 0.266, P = 0.029, 0.039, 0.042 and 0.031, respectively); (3) At week 6 and week 12, there were no statistical differences of the percentages of B-cells and subsets between etanercept group and placebo group of AS patients (P > 0.05); the percentage of CD19(+) B-cells in etanercept group was higher than that in healthy volunteers at week 12 (t = 3.320, P = 0.003).

CONCLUSIONSMisbalance is present in B-cells and some subsets in peripheral blood of active AS patients with peripheral joint involved. B-cells might play an important role in the pathogenesis of AS patients. The high percentage of CD19(+) B-cells in active AS patients cannot be down-regulated after 12-week etanercept treatment.

Adolescent ; Adult ; Antigens, CD19 ; immunology ; B-Lymphocytes ; drug effects ; immunology ; Etanercept ; Female ; Flow Cytometry ; Humans ; Immunoglobulin G ; pharmacology ; therapeutic use ; Immunosuppressive Agents ; pharmacology ; therapeutic use ; Male ; Middle Aged ; Receptors, Tumor Necrosis Factor ; therapeutic use ; Spondylitis, Ankylosing ; drug therapy ; immunology ; Tumor Necrosis Factor Receptor Superfamily, Member 7 ; immunology ; Young Adult

BACKGROUNDAnkylosing spondylitis (AS) is a common inflammatory rheumatic disease which lacks satisfactory treatment so far. Sinomenine (SIN) is an alkaloid and has recently been utilized in treating multiple rheumatic diseases including AS in China, but its exact mechanism remains to be explored. This study investigated the alteration of proteome in peripheral blood mononuclear cells (PBMCs) from AS patients.

METHODSThirty AS patients were enrolled in this study. PBMCs from each AS patient were cultured in medium with or without SIN respectively. Then PBMCs proteins from both groups were separated by two-dimensional electrophoresis (2-DE) and analyzed by mass spectrometry (MS). Two differentially expressed proteins were then chosen to be verified using Western blotting.

RESULTSSeven proteins, including a-synuclein (SNCA), calmodulin (CALM), acidic leucine-rich nuclear phosphoprotein 32 family member A (ANP32A), chloride intracellular channel protein 1 (CLIC1), guanine nucleotide-binding protein G(I)/G(S)/G(T) subunit beta-1 (GNB1), gelsolin (GSN) and histone H2B type 1-M (HISTH2BM) were over-expressed, while coronin- 1A (CORO1A) was under-expressed in the SIN-treated PBMCs. Further bioinformatics search indicated that the changes of SNCA, ANP32A and CLIC1 pertained to apoptosis, while changes of GSN and CORO1A were associated with both apoptosis and inhibition of immunological function. Subsequently GSN and CORO1A were selected to validate by Western blotting and the results were consistent with those of 2-DE.

CONCLUSIONThere were 8 differentially expressed proteins in the SIN-treated PBMCs, which might shed some light on the mechanism of SIN in the treatment of AS.

Adolescent ; Adult ; Blood Proteins ; analysis ; Blotting, Western ; Cells, Cultured ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Leukocytes, Mononuclear ; chemistry ; Male ; Middle Aged ; Morphinans ; pharmacology ; Proteomics ; methods ; Reproducibility of Results ; Spondylitis, Ankylosing ; blood

OBJECTIVETo explore the role of cytokines and keratinocytes in the survival mechanism of mixed auto and allogeneic skin grafting.

METHODSThirty-six SD rats were employed in the study. The rat model with mixed auto and allogeneic skin grafting and mixed human epithelial and lymphocytic culture (MELC) model were established. The change of IL-10 in the serum and the supernatant of the cultured tissue sample from the local wound was observed after the mixed skin grafting in scalded rats. And the role of epithelium in the induction of immunosuppression in vitro was monitored.

RESULTSThe serum IL-10 content in the rats with mixed skin grafting (25.89 +/- 2.82 ng/L) at 7 postoperative day (POD) was evidently higher than that in normal rats (14.20 +/- 2.43 ng/L) (P < 0.05). The IL-10 content in the culture supernatant of rat tissue samples exhibited evident different during 4-14 PODs (P < 0.05-0.01), while which was no difference to that in normal rat on 21st and 28th POD. The inhibiting effects of autologous epithelia and keratinocytes in MELC system were correlated with their dosage. After the adding of autologous keratinocytes to MELC system the cytokines secreted from Th1 could induce the secretion of cytokines from Th2 by IL-10 mediation. This effect could be corrected by the addition of monoclonal antibody of IL-10.

CONCLUSIONThe keratinocytes inlayed in the autoskin during mixed grafting could increase the local IL-10 level by activating Th2 cells, which might be one of the important reasons of the survival of mixed skin grafting.

Animals ; Burns ; immunology ; surgery ; Cytokines ; metabolism ; Giant Cells, Langhans ; cytology ; Graft Survival ; immunology ; Humans ; Interleukin-10 ; metabolism ; Keratinocytes ; cytology ; Lymphocyte Culture Test, Mixed ; Male ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; immunology ; Th2 Cells ; metabolism ; Transplantation, Autologous ; Transplantation, Homologous

Objective To demonstrate the efficiency and safety of the modified reverse controlled antegrade and retrograde subintimal tracking(CART)technique for chronic total occlusion(CTO)recanalization.Methods Thirteen patients with CTO underwent this modified reverse CART technique.Based on the traditional reverse CART,the technology advocated the use of antegrade guidewire Knuckle technology to promote access to the subintimal space.With the use of a strong penetrating and well controllable retrograde guidewire,balloon was inflated in the CTO lesion in favor of advancing the retrograde guidewire through the lesion to reach the proximal true lumen.Results The occlusion site was located in the RCA in 10 patients and in the LAD in 3 patients.Among the 13 lesions,2 had J-CTO score = 1,4 had J-CTO score = 2,6 had J-CTO score = 3,and 1 had J-CTO score=4.Nine cases were considered as the Werner`s Collateral Connection grade 1 and other 4 cases were grade 2.All 13 cases had achieved technical success.1 case needed pericardiocentesis due to collateral branch perforation.Conclusions CTO lesions can be efficiently and safely recanalized using this modified reverse CART technique.

Objective To evaluate the feasibility and efficacy of Guidezilla TM guide extension catheter in retrograde technique for chronic total occlusion (CTO) recanalization. Methods We retrospectively collected 119 CTO cases from the 2nd Cardiology Department of Guangdong General Hospital who had received retrograde percutaneous coronary intervention( PCI). The Guidezilla TMguide extension catheters were applied in 39 cases from October 2015 to November 2016. 80 CTO cases without using the Guidezilla TMguide extension catheter during PCI admitted between January 2015 and October 2015 were collected as the control. Results The overal success rate of the 2 group was 84.0%. The group using the Guidezilla TMguide extension catheter had a higher success rate(100% vs.76.2%,P<0.05).The baseline clinical characteristics of the two groups showed no statistical diff erence. In terms of CTO crossing diffi culty by J-CTO score, patient in the Guidezilla TMguide extension catheter group had higher percentage of diffi cult lesions(35.9% vs.15.0%,P<0.001)and very difficult lesions(61.5% vs.32.5%,P<0.001).Patients in the Guidezilla TMguide extension catheter group had more longer lesions(89.7% vs.72.5%,P=0.035),blunt proximal occlusion(53.8% vs.32.5%,P=0.030)and tortuosity lesions(84.6% vs.63.8%,P=0.020). More cases in the control group needed a second operation(63.8% vs.25.6%,P<0.001)and more ostial/bifurcation lesions(62.5% vs.30.8%,P=0.002).All cases had no target vessel revascularization or in-hospital death. Conclusions Guidezilla TMguide extension catheter is convenient and safe for complicated CTO retrograde PCI.