Objective To investigate the role of TL1A in the generation and differentiation of peripheral blood Th17 in rheumatoid arthritis.Methods The peripheral blood mononuclear cells (PBMCs) of RA were isolated and stimulated with PHA in the presence or absence of TL1A.Naive CD4+ T cells from RA was cultured under Th17-polarizing conditions with or without TL1A.The percentage of Th17 was detected by flow cytometry (FCM) analysis.The DR3 expression on CD4+ T cells from PBMCs of RA patients and healthy controls (HC) were analyzed by using FCM.Data were analyzed with t test and U test.Results TL1A could significantly up-regulate the percentage of Th17 in PBMCs of RA [(7.5±2.3)％ vs (5.2±1.5)％,t=2.647,P＜0.05].Compared to the HC groups,TL1A could significantly induce Th17 differentiation from naive T cells [(37.7±1.9)％ vs (29.5±2.0)％,t=6.455,P＜0.05].The percentage of CD4+DR3+ T cell in PBMCs of RA [(0.56±0.87)％] was significantly higher than that of HC [(0.13±0.04)％,P＜0.05].The percentage of CD4+DR3+ T cell in PBMCs when stimulated with PHA was increased in RA patients,[(4.51±1.34)％ vs (1.11±0.29)％,t=2.915,P＜0.05],but no obvious increase in HC [(0.199±0.104)％ vs 0.072％±0.029)％,t=1.644,P=0.1988].Conclusion TL1A can promote the generation and differentiation of Th17 in the PBMCs of RA,and this effect may be mediated by the binding of TL1A with DR3.

Objective To establish Beagle dogs’model of abducens nerves injury and to observe the clinical therapeu?tic effect of electroacupuncture treatment. Methods Twenty-four Beagle dogs were randomly divided into simple crush group (control group) and crush with electrical stimulation group (experimental group). Cisternal segment of the abducens nerve was given a crush injury, then electrodes were implanted to stimulate the abducens nerve and lateral rectus muscle. Distance between the center of the pupil to medial margin of extraocular adjoin was measured from 1 to 12 weeks after opera?tions. Results All procedures used in the study were well tolerated by Beagle dogs. Electrode implantation to stimulate the lateral rectus muscle and the abducens nerve behind of cavemous sinus was successful. There was no statistical significance of the distance between the two groups from 1 to 2 weeks after operations, and the distance was shorter in experimental group than that in control group from 4 to 12 weeks after operations (P<0.01). Conclusion The animal models established to study electroacupuncture treatment of the injured abducens nerves was successful. Electroacupuncture can promote the re?covery of the injured abducens nerves obviously.

Objective To detect the changes of number and function of endothelial progenitor cells (EPCs) in gld.apoE-/-C57BL/6 mice,and to investigate whether premature atherosclerosis of gld.apoE-/-C57BL/6 mice was mediated by the dysfunction of these EPCs.Methods EPCs were isolated from peripheral blood and bone marrow of four types of C57BL/6 female mice:gld+/+apoE+/+,gld,apoE-/-and gld.apoE-/-.The percentage of EPCs was analyzed by FACS as CD11b-Sca-1+CD309+ cells.The attached cells were stained with DiI labeled acetylated low-density lipoprotein (DiI-ac-LDL) and FITC-labeled Ulex Europaeus agglutinin 1 (FITC-UEA-1) double staining to determine their phagocytic function.The adherent function of EPCs was determined by calculating the number of re-cultured EPCs.The capacity of EPCs to form the cavity structure was detected by calculating the number of the formed vascular-like structure.Results The percentage of circulating EPCs was significantly decreased in the gld.apoE-/-group (0.012±0.002)％ compared to the gld+/+ apoE+/+ group [(0.039±0.005)％,P＜0.01],the gld group [(0.025±0.001)％,P＜0.05],and the apoE-/-group [(0.028±0.002)％,P＜0.01].The percentage of bone marrow derived EPCs was decreased in the gld.apoE-/-group (0.12±0.01)％ compared to the gld+/+apoE+/+ group [(0.42±0.05)％,P＜0.05].The percentage of DiI-acLDL and FITC-UEA-1 double positive cells was decreased in the gld.apoE-/-group [(59.2±2.1)％] compared to the gld+/+apoE+/+ group [(87.5±3.0)％,P＜0.01],and the gld group [(84.2±6.0)％,P＜0.01] ; the adherent function of EPCs was impaired in the gld.apoE-/-group [(50.0±5.3)％] compared to the gld+/+ apoE+/+ group [(86.0±7.3)％,P＜0.01],the gld group [(73.0±1.0)％,P＜0.01],and the apoE-/-group [(65.0±4.0)％,P＜0.05] respectively.The capacity of EPCs to form the cavity structure was decreased in the gld.apoE-/-group (4.0±0.3) compared to the gld+/+apoE+/+ group (12.0±1.4,P＜0.01).Conclusion The number of EPCs is decreased in the gld apoE-/-C57BL/6 mice,the adhesion,phagocytosis and angiogenic function of EPCs in the bone marrow are impaired,which may be one of the causes of lupus with atherosclerosis.

ObjectiveTo investigate the number of γδT cells in the peripheral blood from patients with systemic lupus erythematosus(SLE) and their correlation to disease activity.MethodsγδT cells were detected in the peripheral blood from 42 SLE patients and 20 normal controls by flow cytometry.Anniex-V/Pl double-staining flow cytometry was employed to observe the proportion of the apoptotic γδ and CD3+ T cells in 6 SLE patients with active disease and 6 normal controls,respectively.The levels of plasma anti-nuclear antibody and anti-dsDNA antibody were determined by using enzyme-linked immunosorbent assay.Data were analyzed with t test and Pearson's correlation test.ResultsThe percentages of γδT cells were remarkably down-regulated in SLE patients [(3.0±1.8)％ ] with active disease compared with that of those patients with inactive[(5.3±3.0)％] disease and normal controls [(6.8±2.8)％](t=-3.071 and -5.913 respectively,both P＜0.01 ).The absolute number of γδT cells decreased significantly in SLE patients with active disease[ ( 1.7± 1.6)× 107/L ] than those with inactive SLE [ (5.3±3.6)× 107/L ] (t=-3.292,P＜0.01 ),and both were lower than the normal controls [ (10.1±5.0)×l 07/L] (t=-7.247 and -2.905 respectively,both P＜0.01 ).There was a negative correlation between systemic lupus erythematosus disease activity index (SLEDAI) andT cell counts in 30 SLE patients with active disease (r=-0.365,P=0.047).γδT cell percentage (r=-0.336,P=0.030) and counts (r=-0.410,P=0.007) were both inversely correlated with the erythrocyte sedimentation rate,but positive correlation were found between hemoglobulln and γδT cell counts (r=0.409,P=0.007).The apoptosis ofγδT cells in SLE patients was more common than in normal controls(t=2.886,P＜0.05 ).The number of apoptotic γδT cells was higher than that of CD3+ T cells in SLE patients (t=2.952,P＜0.05 ).Conclusion γδT cells of the peripheral blood of SLE patients are down-regulated partially due to excessive apoptosis,which may correlate with the disease activity.

ObjectiveTo analyze the correlated clinical significance by testing the serum tumor necrosis factor like ligand-1A(TL1A) levels of patients with rheumatoid arthritis(RA). MethodsThe serum from 100 RA patients and 50 healthy controls were selected. The serum TL1A concentrations of the two groups were detected by ELISA. The correlations between serum TL1A levels and clinical features or laboratory examinations of RA patients were investigated. Mann-Whitney U test, t-test, x2 test and Spearman correlation analysis were used for statistical analysis. ResultsThe levels of serum TL1A in RA patients [(959±1146) pg/ml]were significantly higher than those of the healthy controls[(529±154) pg/ml, t=3.683,P＜0.01]. The levels of serum TL1A in RA patients with positive rheumatoid factor(RF),occult rheumatoid factor immunoglobulin G (HRF-IgG) and anti-cyclic citrullinated peptide antibodies (anti-CCP)[(962±1043 ), ( 833±1104 ), (908±1115 ) pg/ml, respectively]were significantly increased when compared to those in RF, HRF-IgG and anti-CCP negative RA patients[(628±r343), (576±134),(628±401) pg/ml, respectively, t=3.224, 1.317, 1.003; P＜0.05]. The positive rates of RF, HRF-IgG and anti-CCP in the TL1A positive group(90％, 42％, 85％) were significantly higher than those of the TL1A negative group (56％, 11％,33％) (x2=-0.372, -2.402, -2.774; P＜0.05). ConclusionThe level of serum TL1A is significantly elevated in RA and it is significantly associated with auto-antibodies in RA including RF, HRF-lgG and antiCCP antibody, which may be a factor for unfavorable prognosis of RA patients.

Objective To test serum decoy receptor 3 (DcR3) concentration in patients with rheumatoid arthritis (RA),and analyze its clinical significance.Methods The serum from 180 RA patients,29 osteoarthritis (OA) patients,60 systemic lupus erythematosus (SLE) patients,30 Sj(o)gren's syndrome (SS) patients and 100 healthy donators were selected.The serum DcR3 concentrations of the two groups were detected by enzyme-linked immunosorbent assay (ELISA).The correlation between serum DcR3 levels and clinical features of patients with RA were investigated.T test,x2 test and Spearman's correlation analysis were used for statistical analysis with software SPSS 16.0.Results Serum DcR3 levels in RA patients [(202±261)ng/ml] were significantly higher than those of the healthy controls [(32±31) ng/ml,t=5.33,P＜0.01],OA patients [(90±33)ng/ml,t=1.998,P＜0.01],SLE patients [(41±56)ng/ml,t=3.19,P＜0.01],and SS patients [(25±31) ng/ml,t=2.50,P＜0.01].The DcR3 cut value was 45.86 ng/ml for the diagnoses of RA,the sensitivity and specificity was 0.991 and 0.765 respectively.The levels of serum DcR3 in RA patients were related with rheumatoid factor (RF) IgM,RF-IgG,anti-cyclic citrullinated peptide antibodies (anti-CCP) and C3.Conclusion Serum DcR3 level is significantly elevated in RA and it is significantly associated with immunological index of RA including RF-IgM,RF-IgG,C3 and anti-CCP antibody,which may be a factor for unfavorable prognosis of RA patients.

ObjectiveTo investigate the role of magnetic resonance imaging (MRI) in treatment efficacy evaluation of polymyositis and dermatomyositis.MethodsFifteen patients with polymyositis and dermatomyositis underwent MRI of thigh were included.Scores of MRI signal intensity of the diseased muscle of every patient were compared before and after treatment and the correlation between serum creatinkinase (CK) level and muscle strength grade were also compared.Correlations between muscle strength grade and MRI score,as well as muscle strength grade and creatinkinase level were analyzed.Comparisons between groups were tested by t test,and the relationship between muscle strength and clinical data was analyzed by Pearson's correlation analysis.ResultsThe signal score of MRI was counted before and after therapy(2.37±0.62,1.30±0.28,respectively,P＜0.05),and CK level[(3841±3175),(549±338) U/L,respectively,P＜0.05] and muscle strength (15.1 ±2.4,18.1 ±0.9,respectively,P＜0.05) were assessed at the same time.Muscle strength grade was associated with signal score of MRI and serum CK level,there was a strong correlation between muscle strength grade and signal score of MRI(r=-0.890,P＜0.05 ).ConclusionMRI may be a useful tool for clinical efficacy evaluation in patients with polymyositis and dermatomyositis.

Objective To investigate the expression of FcγRⅡb on peripheral B cells and its clinical significance in primary Sj(o)gren's syndrome (pSS).Methods FcγRⅡb expression on peripheral B cells from 19 pSS patients and 15 healthy controls was examined by flow cytometry.The levels of serum anti-SSA and SSB antibodies were determined using enzyme-linked immunosorbent assay (ELISA).Data were analyzed with t test,one-way ANOVA,SNK-q test and Pearson's correlation test.Results The percentage of memory CD19+CD27+ B cell subpopulation was significantly lower in pSS patients [ (20.8±2.7)％] when compared to normal controls [(37.8±2.2)％ ](t=-4.002,P＜0.01).The level of expression of FcγRⅡb in active pSS memory CD19+CD27+ B cells [ MFI(74±8)] was significantly reduced when compared to inactive [ MFI( 132±11)] and normal controls [ MFI (139±12)] (F=10.699,P＜0.01).The level of expression of FcγRⅡb on memory CD19+CD27+ B cells from pSS patients was inversely correlated with Sj(o)gren's syndrome disease activity index (SSDAI) (r=-0.744,P=0.0003 ).pSS patients with the serum anti-SSA/SSB antibodies positive group [ MFI(75+3),(48±7)] displayed a lower expression of FcγRⅡb on memory CD19+CD27+ B cell than in patients with the serum anti-SSA/SSB antibodies negative group [MFI( 122±11),(108±9)] (t=-4.336 and -3.776 respectively,the P value of both tests were less than 0.01).The level of expression of FcγRⅡb in the memory CD19+CD27+ B cells of patients with active pSS was inversely correlated with anti-SSA antibody titers (r=-0.685,P=0.014).Conclusion The expression of FcγRⅡb on peripheral memory B cells from active pSS patients is inversely correlated with SSDAI and is also inversely associated with anti-SSA antibody levels.Decreased expression of FcγRⅡb might play an important role in the pathogenesis of pSS.

Objective To investigate the dose-response relationship of the treatment temperatures and heating time on human cervical carcinoma hela cells,aiming at providing experimental evidences for clinical hy-perthermia. Methods Hela cells were heated at 37 ～ 70 ℃ in temperature-controlled water baths, the tempera-ture was divided into nine groups,each time was divided into eight subgroups (1 ～ 30 min). The morphology changes of cells after hyperthermia were detected by inverted microscope. Proliferation rates were measured by MTT colorimetric assay. The apoptesis rates were determined by flow eytometric analyse. The levels of prolifera-ring cell nuclear antigen (PCNA) were measured with immunohistochemistry. Results lnereaseing the heating time at the same temperature, or increaseing heating temperature at the same time, the cell proliferation, survival rates and PCNA expression decreased. There was no significant morphological change about cells ,but have small amount of apoptosis and a direct role of the suppression and destruction at 41 ℃ and 43 ℃ group. A large num-ber of cells shrinked to round and a major role for apoptosis at 46℃ group. Cell necrosis was major role at 50 ℃and 55 ℃ group. More than 55 ℃ for necrotic cells. Conclusion With the increase of heating temperature and heating time, its treatment of Hela cells gradually enhance. So combining dose-effect relationship of hyperthermia temperature and time can reach the best therapeutic effects.

Objective To investigate the effect of interleukin(IL)-21 and dexamethasone(Dex)treatment on the percentage of follicular helper T cells(Tfh)in patients with systemic lupus erythematosus (SLE).Methods Peripheral blood mononuclear cells(PBMCs)obtained from 11 SLE patients were cultured in 96 well plates at 106/well with medium as the control,and with rIL-21(200 ng/μl)or rIL-21(200 ng/μl)+Dex(1×10-6 mol/L)for 24 and 72 hours as the study groups.Another 20 samples were collected and incubated with different concentrations of Dex(0,5×10-7 mol/L,1×10-6 mol/L)for 24 hours.After labeling with CD4,PD1 and CXCR5 monoclonal antibodies,the proportions of Tfh cells were assessed by flow cytometry.ELISA was used to detect supernatant antinuclear antibody(ANA)levels.Differences between groups were analyzed by paired t test.Results The percentages of Tfb cells were elevated when incubated with rIL-21 for 24 h and 72 h[24 h(4.3±1.2)％,(5.9±2.4)％;72 h(5.6±4.0)％,(8.0±5.6)％;t=3.48 and 3.05 respectively,P=0.007 and 0.012].Compared with those in the rIL-21 group,the percentages of Tfh cells in the rIL-21+Dex group were decreased(t=4.70 and 2.78,P=0.001 and 0.019).Tfh cells were decreased ina dose-dependent manner when treated with Dex[24 h(4.2±1.3)％;72 h(6.2±4.4)％;t=3.37 and 2.26,P=0.003 and 0.036].There was no difference of supe(rn)atant ANA levels among those groups.Conclusion IL-21 can promote the proliferation of Tfh cells,while Dex inhibits Tfh cell growth and the inhibition effect is dose dependent.Tfh cells may represent a new target for the treatment of SLE.