Objective:To investigate the specific stimulation of the B cell epitope peptides of Mycobacterium tuberculosis antigen (Mtb-Ag) on human peripheral γδ T cell proliferation.Methods: We selected the sequences of B cell epitope peptide from Mtb-Ag that were reported in literature and T cell epitope peptide that recently identified in this laboratory to synthesize six peptides of B cell epitopes (BP1-BP6) and two peptides of γδ T cell epitopes (TP14,TP15).The 24-well culture plates were coated with these peptides.The PBMCs were isolated from peripheral blood of healthy individuals and stained with CFSE,followed cultured for 12 days in the IL-2 containing medium.Mtb heat resistant antigen ( Mtb-HAg ) group as positive control and IL-2 only group as negative control.The percentages and proliferation index of γδ T cells were determined by flow cytometry.Results: By using Wilcoxon signed rank test for paired comparison of negative control group,the percentages of γδ T cells in cultured PBMCs with BP2,BP4 and TP14, TP15 and Mtb-HAg increased significantly in 14 samples (P<0.05);and the proliferation index of γδT cell in cultured PBMCs with BP2,BP4,BP5,BP6 and TP14,TP15 increased significantly in 7 samples (P<0.05).Conclusion: Taken together,the B cell epitope peptides from Mtb Antigens are capable of stimulating the γδ T cell proliferation specifically in vitro.Although there was individual difference inγδT cell proliferative response to B cell epitope peptides,these results strongly suggest the B cell epitope peptides also can specifically trigger the TCR ofγδT cells.

Objective To study the morphological changes in peripheral blood cells and bone marrow cells of the patients suffering from mustard gas poisoning and their relationship with the degree of poisoning. Method The peripheral blood cells and marrow cells were examined morphologically after mustard gas poisoning. Results The total WBC count was reduced progressively in 44 patients, among them 10 patients showed granulocytopenia or agranulocytosis. Severe injury to eosinophils was seen also in the early period, and the percentage and absolute value of lymphocytes were significantly lowered too. However, platelets were not significantly influenced. The red cell count and hemoglobin level were elevated because of hemoconcentration. Blood marrow cells showed marked morphological changes. WBC became swollen with appearance of poisonous granules and vacuoles in their cytoplasm. Deformed lymphocytes (LY) could be found in the marrow cells of all the patients, mainly in the form of immature lymphocytes. It was shown that mustard gas produced serious damage to the hematopoitic cells of the bone marrow in the early period. Conclusions The examination of pheripheral blood and bone marrow cells is an important measure to determine the degree of mustard gas poisoning. The decrease in the absolute value of lymophocyte count and the time of its recovery, and degree of inhibition of hematopoiesis are closely related to the degree of mustard gas poisoning.

Objective To detect the frequency of peripheral blood CD4+ CD25high Tr and CD4+CD25lowT cells and expression of programmed death-1 ( PD-1 ) on their surface in the patients with systemic lupus erythematosus(SLE) and their clinical signifcance is analyzed.Methods The expression of PD-1 on the CD4+ CD25highTr and CD4+CD25lowT cells was examined in patients with 33 active SLE,18 inactive SLE and 38 healthy controls(HC) by flow cytometry.Clinical manifestations and laboratory findings of SLE were collected.Patients were divided into two groups according to their disease activity.SLE disease activity index(SLEDAI) score≥10 was defined as high disease activity and ＜10 as inactivity.The percentage of CD4+ CD25highTr and CD4+CD25lowT cells and proportions of expression of PD-1 on their surface were compared between not only inactive or active SLE patients and healthy controls (HC),but also between patients with lupus nephritis and without lupus nephritis.Correlation with clincal manifestations and laboratory findings was analyzed.Results (1)The proportions of CD4+CD25highTr and CD4+CD25low T cells were significantly decreased in active and inactive SLE patients as compared with HC.(2)The percentage of CD4+ CD25lowPD-1 +Tr and in active and inactive SLE patients were higher than HC.The proportions of CD4+CD25lowPD-1+ T cells were significantly increased in HC and inactive SLE patients as compared with inactive SLE patients.(3) The proportions of CD4+CD25highPD-1+Tr in SLE patients with nephritis were higher than those in patients without nephritis.But the percentage of CD4+ CD25 lowPD-1 + T cells was significantly decreased in SLE patients with nephritis as compared those without nephritis.(4)A positive correlation was observed for proportions of CD4+CD25highPD-1 + T cells with SLEDAI score.Proportions of CD4+ CD25high Tr,CD4+ CD25low PD-1 + T cells was inversely correlated with SLEDAI score.Conclusion The aberrations of proportions of CD4+CD25highpD-1+,CD4+CD25lowPD-1+,CD4+CD25high and CD4+CD25low T cells were observed in patients with SLE.The abnormality of PD-1 and PD-L1 pathway may play an important role in the function and number of Tr.

BACKGROUND:No method is ideal for treating traumatic avascular necrosis of talus up to now.Extracorporeal shock wave therapy(ESWT)is a micro-traumatic,simple,and effective method to treat musculoskeletal diseases;however,the therapeutic effect on necrosis of talus needs to be further studied.OBJECTIVE:To evaluate the therapeutic effect of liquid-electric extracorporeal shock wave on traumatic avascular necrosis of talus,and to explore new treatments of traumatic avascular necrosis of talus METHODS:A total of 34 patients with traumatic avascular necrosis of talus were selected from the Affiliated Hospital of Medical College of Chinese Armed Police Force from September 2004 to June 2009.The patients were randomly divided into ESWT and control groups,with 17 patients per group All patients were treated with pain point positioning combined with surface X-ray localization,theworking voltage of 8-10 kV,energyflowdensity of 0.12-0.16mJ/mm2,impact frequency of 40-50 times/min,and impact of 800-1000 times,once a week,for 3-5 cycles.Pain was evaluated with VAS before and after treatment,function of ankle was evaluated with AOFAS standards,and MRI of ankle was re-checked at 18 months after treatment to compare necrotic area before and after treatment.RESULTS AND CONCLUSION:VAS pain,function of ankle,and necrotic area of ankle in the ESWT group were significantly improved compared to those in the control group at 18 months after treatment(P<0.01).Activity of one case in the control group was limited by severe pain due to traumatic arthritis in the first 15 weeks after ankle arthrodesis surgery.This suggested that significant effect and fewer complications,for treating traumatic avascular necrosis of talus.

OBJECTIVE:To study the effects of nimesulide combined with oxaliplatin on microvessel density and immune func-tion of esophageal cancer model rats. METHODS:48 rats were selected to establish esophageal cancer model and then randomly di-vided into model group,oxaliplatin group,nimesulide group and combination group,with 12 rats in each group. 1 d after model-ing,model group was given 5% Glucose injection 1 mL via tail vein,3 times a week,and constant volume of Sodium carboxy-methylcellulose solution,once a week. Oxaliplatin group was given oxaliplatin 13.6 mg/kg via tail vein,3 times a week. Nimesu-lide group was given nimesulide 20 mg/kg intragastrically,once a week. Combination group was given oxaliplatin and nimesulide with same usage as above. The administration lasted for 8 weeks. The cancer growth,microvessel density of cancer tissue,peripher-al blood immune function index were observed in four groups. RESULTS:Compared with model group,cancer size,cancer weight,integral absorbance of cancer and positive vessel density were all decreased in treatment groups(P<0.05). The number of peripheral blood CD3+,CD4+T cells and CD4+/CD8+were all decreased in oxaliplatin group and combination group,while the num-ber of CD8+T cells was decreased(P<0.05). Compared with nimesulide group,cancer size,cancer weight,integral absorbance of cancer,positive vessel density,the number of peripheral blood CD3+,CD4+T cells and CD4+/CD8+ were all decreased in oxaliplat-in group and combination group,while tumor inhibition rate and the number of CD8+T cells were all increased(P<0.05);combi-nation group was more significant than oxaliplatin group (P<0.05). CONCLUSIONS:Oxaliplatin combined with nimesulide can effectively inhibit cancer growth of esophageal cancer,the mechanism of which may be associated with inhibiting angiogenesis of tumor tissue and strengthening immune function.

Objective To investigate the carotid atherosclerosis(AS) in hemodialysis(HD) patients.Methods Clinical index was measured and intma-media thickness(IMT) of extracranial common carotid artery and presence of atherosclerotic plaques were determined by high-resolution B-mode ultrasonography.Results The age,disease duration,HD duration,triglyceride(TG),low density lipoprotein(LDL),serum C-reactive protein(CRP),mean carotid artery IMT in HD with atherosclerotic plaque were significantly higher than HD without atherosclerotic plaque patients,however,the serum albumin(Alb) was significantly decreased.Mean carotid artery IMT is related to the disease duration,Alb,TG,LDL,CRP in HD patients.Conclusion The CRP,mean carotid artery IMT were increased in HD with atherosclerotic plaque.Mean carotid artery IMT is related to the disease duration,Alb,TG,LDL,CRP in HD patients.To improve malnutrition and decrease micro-inflammation,serum lipid might reduce the rate of AS in HD.

Objective:To observe the stimulating effect of C main Peptide purified from Mycobacterium tuberculosis peptide antigens(Mtb Ag) on human ??T cells, and the effect of C main peptide on the Mtb Ag activated ??T cells in vitro. Methods:C main peptide was used to stimulate fresh ??T cells of normal subjects in different doses for 10 d and the responded cells were phenotyped by flow cytometry. Furthermore,C main peptide was used to restimulate the activated ??T cells ,and then analyse the expression of early activation marker molecule CD69 in ??T cells by flow cytometry and the activity of proliferation of ??T cells by MTT assay. Results:C main peptide could stimulate the proliferation of fresh ??T cells predominantly, and it also could promote the expression of CD69 in activated ??T cells and enhance its activity of proliferation. Conclusion:C main peptide purified from Mtb Ag is a kind of specific stimulator of human ??T cell in vitro.

Objective The aim of this study is to examine the expressions of Toll like receptor (TLR) 7 and TLR9 in the peripheral blood B lymphocytes of SLE patients and to analyze the correlation between TLR7/9 and clinical parameters. Methods lntracellular expression of TLR7/9 in the peripheral blood CD19+Blymphocytes was analyzed in 50 SLE patients and 30 healthy controls by flow cytometry. The difference of intracellular TLR7/9 expression levels in two groups was compared. Furthermore,the correlation between TLR7/9 expression and clinical parameters such as ESR, CRP, complement 3 (C3), complement 4 (CA), the level of serum IgG, anti-double stranded DNA antibody, anti-nuclear antibodies, SLEDAI score and urine protein excretion level, were analyzed. Results Compared with healthy subjects, the proportion of B cells expressing TLR7 and TLR9 was higher among SLE patients. Positive correlation was observed between TLR7 expression levels and clinical measurement of the SLEDAI and ESR. Negative correlation was observed between TLR7 expression levels and serum C3 levels. Positive correlation was observed between TLR9 expression levels and SLEDAI scores. Negative correlation was observed between TLR9 expression levels and serum C3 levels. Conclusion TLR7 and TLR9 expression is increased in the peripheral blood B cells of SLE patients, and correlates well with clinical parameters.

Objective To explore the roles of PKCθ(protein kinase Cθ)signaling pathway on the activation,proliferation and TH1/TH2 cytokines production of the γδT lymphocytes stimulated by Mycobacterium tuberculosis antigen(Mtb-Ag) in vitro.Methods Peripheral blood mononuclear cells(PBMC)were pretreated with or without Rottlerin at 5.0 μmol/L,and stimulated with Mtb-Ag and cultured in rhIL-2 containing medium.After different time of culture,activation molecules and cytokines production of γδT cells were measured by flow cytometry.The proliferation proportion and the percentage of each generation of γδT cells were determined by carboxylfluoreseein diacetate succinmidyl ester(CFSE)labeling technique and flow cytometry.Results After 3d of stimulation with Mtb-Ag,the expression rates of CD69 and CD25 of γδT cells were 46.2%and 45.6%,respectively.Pretreatment of 5.0 μmol/L Rottlerin markedly inhibited the both expressions of CD69 and CD25 in γδT cells(P＜0.01).After stimulation and expansion in vitro for 5,10,and 15 d,the percentages of the γδT cell were 9.6%,54.6%and 82.4%,respectively.There was a few γδT cells in propagation on the 5th day of culture,and almost all γδT cell divisions were above 6 generations on the 10th and 15th day of culture.Pretreatment of the Rotflerin significantly suppressed the γδT cell proliferation,but after 10 d culture,there were still a few parts of γδT cells in propagation.Meanwhile,after 7,14,and 21d of culture,upon stimulation with PMA+Ionomycin,the IFN-γ producing-γδT cells were about 80%at all times.But only after 21d culture,IL-4-producing γδT cells was 2.6%.,The percentage of IFN-γ producing γδT cells markedly reduced in Rottlerin group(P＜0.01).IL-4 secretion of the γδT cells was almost completely blocked.Conclusion PKCO signal pathway involves in activation,proliferation and differentiation of the γδT lymphocytes stimulated by Mtb-Ag.

85%. The concentrated MCS in different amount was added to the IFN-?(100 pg/mL) and LPS (10 ng/mL) enriched culture media. The IL-12 production by monocytes was determined by the enzyme- linked immunosorbent assay(ELISA).The expression of CD14 and CD1a was analyzed by flow cytometry 5 days after the monocytes were co-cultured with MCS. Results The production of monocytic IL-12 was down-regulated by MCS in a dose dependent manner. The amount of IL-12 from monocytes decreased along with an increased dose (25-100?L) of MCS applied in the reaction. It was also observed that the differentiation from CD14 expressing monocytes to CD1a dendritic cells was impaired by MCS. The ability of MCS to inhibit the production of IL-12 by monocytes and to suppress the differentiation of monocytes to dendritic cells in vitro could be disrupted by PD98059,an ERK specific inhibitor. Conclusions MCS appears to inhibit IL-12p40 production by monocytes and inhibit differentiation of monocytes in vitro via secretion of ERK stimulating factor. The inhibitory factors in MCS and their chemical natures need further research.