The psychological adjustment of disabled athletes generally experienced five times,psychosocial rehabilitation of disabled athletes should be targeted for education and other cognitive therapy.To find out the psychological features of disabled athletes,take health education such as the psycho-social rehabilitation is of far-reaching significance to help disabled athletes restoring good mental state,raising the level of mental health,reconstructing social behavior,and re-entrying society as soon as.

AIM: To investigate the expression kinetics of PD-L1 and PD-L2 on the surface of the resting and activated B/T cells as well as monocytes from healthy human peripheral blood.METHODS: Fluorescent antibody staining together with flow cytometry were used to detect the percentages of the resting as well as the activated B cells and T cells that expressed PD-L1 and PD-L2.Meanwhile the percentages of the resting and activated monocytes that expressed PD-L2 were determined.RESULTS: Both resting B cells and T cells did not express PD-L1 on their surface,however PD-L1 expression was significantly up-regulated on the surface of the activated B cells after 6 h stimulation with LPS or pokeweed mitogen(PWM),and the percentages of B cells that expressed PD-L1 reached a plateau at 24 h,which were(46.26?10.71)% with LPS and(43.67?6.14)% with PWM stimulation,respectively.No markedly change of PD-L1 expression on the surface of the activated T cells after stimulation with LPS was observed,but upregulation of PD-L1 expression was observed when stimulation with PWM.The percentages of T cells that expressed PD-L1 reached a plateau at 24 h,which was(25.42?9.23)%.PD-L2 expression was not found on the resting as well as the activated B cells and T cells.In addition,the resting monocytes did not express PD-L2.Combination of INF-? plus LPS markedly induced the PD-L2 expression,and the percentages of monocytes that expressed PD-L2 reached a peak at 48 h,which was(28.70?14.22)%.CONCLUSION: The activated lymphocytes only express PD-L1,reaching a plateau at 24 h.PD-L2 is expressed on the surface of the activated monocytes,reaching a peak at 48 h.

AIM: To investigate the amount and patterns of expressing CD69 , IL-4 and IFN-? on TCRV?24 +NKT cells, and compare with that of CD3 +T cells from human peripheral blood in response to in vitro stimulation. METHODS: The whole blood was stained with three-color immunofluorescence directly or after cultured with PDB+ionomycin(Ion) for 6 h, then the mononuclear cells were separated by lysing red blood cells. The expression rates of CD69, IL-4 and IFN-? on TCRV?24 +NKT cells and CD3 +T cells were estimated by flow cytometer. RESULTS: As a proportion of mature T cells, the ratio of TCRV?24 + NKT cells to CD3 +T cells was about(1.34?0.42)%. The expression rates of CD69 on TCRV?24 + NKT cells and CD3 +T cells in response to PDB + Ion for 6 h were (96.71?1.33)% and (98.60?0.47)%, respectively, while the ratio were (11.47?2.86)% and (1.07?0.45)% in the unstimulated group, and there were significant difference between them. The expression rates of IL-4 and IFN-? on TCRV?24 +NKT cells stimulated with PDB+Ion for 6 h were (48.62?2.44)% and (46.65?8.91)%, respectively ,which were significantly higher than that of unstimulated group [(31.57?3.31)%, (13.45?6.29)%] and that of stimulated CD3 +T cells, though the expression rates on stimulated CD3 +T cells were significantly higher than that of unstimulated CD3 +T cells. CONCLUSIONS: There is small amount of NKT cells in adult human peripheral blood. The expression rates of IFN-? and IL-4 on these lymphocytes are higher than CD3 +T cells, suggesting that NKT cells are important immunomodulatory cells in special microvironments.

AIM: To investigate the effect of berberine (Ber) on the activation and proliferation of T lymphocytes and its mechanism of action. METHODS: Whole peripheral blood from normal subjects was stimulated with phytohemagglutinin (PHA) or phorbol ester (PDB) plus ionomycin (Ion) and the expression levels of CD69 and CD25 were evaluated with flow cytometry after the staining with appropriate fluorescent monoclonal antibody. The distribution of cell cycles was analyzed by propidium iodide staining and dead cells by 7-aminoactinomycin live staining. RESULTS: 100 ?mol/L and 50 ?mol/L of Ber had significant inhibition of the expression of CD69 on T cells stimulated with PDB plus Ion or PHA, while effect of 25 ?mol/L Ber was not significant. And as time of action extended, the extent of inhibition decreased. For the expression of CD25, Ber at the concentrations as above all exerted significant inhibitory effect in a dose-dependent manner. Moreover, Ber could block lymphocytes cell cycle progression from G_0/G_1 phase to S and G_2/M phase without phase specificity. Besides, live staining analysis revealed that Ber did not have significant cytotoxicity on lymphocytes. CONCLUSIONS: Ber significantly inhibits the expression of early and mid activation antigens of T cells and also blocks the progression of lymphocytes cell cycles. These results suggest that Ber exerts immunosuppression effect through inhibiting the activation and proliferation of T cells.

AIM: To investigate the details of age-related changes of T lymphocytes in order to seek for sensitive biomarker for immunosenesence. METHODS: Heparin anticoagulated venous blood was collected freshly from young (20-35 years) and elderly (50-75 years) volunteers and three or four color immunofluorescence staining was performed. The nucleated cells were acquired and the phenotypes of T lymphocyte subpopulations were analyzed with flow cytometry. RESULTS: There was no significant difference in percentages of pan-T (CD3 +), helper T (CD4 +) and cytotoxic (CD8 +) T subsets between young and elderly, whereas the density of CD3 molecule (MFI) on T cells in elderly group decreased significantly. It was also found that the rates of CD44 + and CD62L + T cell subsets in young group did not have statistical difference from elderly. However,the rates of CD95 + pan-T, helper T and cytotoxic T subsets of elderly group were all markedly higher than that in young group. CONCLUSIONS: The relative rates of T cell and its subsets displayed no age-related changes while the density of CD3 was down-regulated during aging in these groups investigated. Moreover, the expression percentage of CD95 (Fas) on T cells increased as aging, suggesting that it is a potential biomarker for evaluating immunosenescence.

AIM: To construct the mammalian cell expression vector for enhanced green fluorescent protein (EGFP) and HLA-A*0201 fusion protein and analyze its expression and subcellular localization in the transfected K562 cells. METHODS: The HLA-A*0201 cDNA was cloned by RT-PCR and the gene was inserted into pEGFP-N1 to construct a vector for the fusion protein. The expression of the fusion protein in K562 cells transfected with the vector was evaluated by flow cytometry and its subcellular localization was investigated by confocal microscopy. RESULTS: The full-length encoding region of HLA-A*0201 cDNA was cloned from two HLA-A2 positive donors and the expression vector for the HLA-A*0201-EGFP fusion protein was constructed by PCR using a primer pair to introduce a Kozak sequence before ATG and the stop codon was deleted. Five hours after K562 cells was transfected with the vector, the expression percentages of HLA-A*0201 and EGFP were 25.12?2.26 and 27.37?3.59, respectively and no significant increase was observed after 24 h. The fusion protein was predominantly located on the membrane with low level distribution within the cells. In contrast, no HLA-A*0201 but only EGFP was detected in the empty vector transfected K562 cells and the EGFP was dispersed within the cells. CONCLUSIONS: The expression vector for HLA-A*0201-EGFP fusion protein was constructed and the fusion protein expressed in K562 cells was primarily distributed on the membrane. The results suggest that the transfected K562 cells are potential antigen-presenting cells.

Objective: To study the therapeutic effect of secondary lymphoid tissue chemokine (SLC) combined with CpG oligodeoxynucleotide (CpG-ODN) in treatment of implanted mouse melanoma and the possible mechanism. Methods: SLC-Fc fusion protein was prepared and its chemotaxis of lymphocytes was detected by chemotaxis assay. Implanted melanoma mouse models were established and randomly divided into 4 groups: control group, SLC-Fc group, CpG-ODN group, and SLC-Fc+CpG-ODN group. The growth of implanted tumors in each group was observed after treatment. Subtype and infiltration of lymphocytes in implanted tumor tissues were examined by flow cytometry. Results: SLC-Fc protein was successfully prepared, and it dose-dependently attracted lymphocytes (0.03, 0.3, and 3 μg/L). Intra-tumor injection SLC-Fc and CpG-ODN alone or in combination significantly inhibited growth of B16-implanted tumors. Tumor size in SLC-Fc+CpG-ODN group was significantly smaller than that in control group (P<0.01), and animals in SLC-Fc+CpG-ODN group survived longer. Tumor-infiltrated CD4~+ T, CD~8+ T, and dendritic cells (DCs) in SLC-Fc+CpG-ODN group were markedly increased as compared with those in control group (P<0.05), and tumor draining lymph nodes were dramatically enlarged. Conclusion: SLC combined with CpG-ODN can inhibit the growth of implanted melanoma by attracting CD4~+ T and CD8~+ T and promoting proliferation of DCs.

According to the level of selenium (Se) and methionine (Met) in diet, Wistar rats were randomly divided into 3 groups, group A (normal Se and normal Met); group B (Low Se and normal Met); group C (low Se and low Met). The animals were sacrificed after 8-week feeding. The results showed that the alteration of myocardial ultrastructure was slight in group B. Decreased activities of cytochrome oxidase and succinate dehydrogcnase of myocardial mitochonria and decreased activities of GSH-Px in the blood, heart and liver in group B were observed, as compared with those in group A. But the level of TBA was higher than that of group A.The above mentioned changes in group C were extremely apparent than group A or B, indicating both Se and Met deficiency in diet may concern the etiology of Keshan disease.

Aim To explore the influence of metformin(a first-line drug for type 2 diabetes) on ATP-induced inflammasome activation and the release of interleukin-1β(IL-1β) by LPS-activated peritoneal macrophages, a commonly-used inflammatory cell model.Methods Peritoneal macrophages were elicited by intraperitoneal injection of 30 g·L-1 thioglycollate into C57BL/6 mice.Inflammasome was activated and cell pyroptosis was induced by LPS plus ATP treatment, and the pyroptotic cells were calculated after propidium iodide(PI) staining.The protein levels of IL-1β and caspase-1 expressed in the cells and released from them into the supernatant were evaluated by Western blot.Immunofluorescent microscopy was recruited to detect the subcellular distribution and fluorescent intensity of the purinergic P2X7 receptor(P2X7R).Results Metformin per se did not induce pyroptosis in LPS-activated peritoneal macrophages, but it significantly and dose-dependently increased cell pyroptosis induced by ATP treatment.At protein levels, maturated IL-1β(17 ku) could not be released from the cells upon single LPS or LPS plus metformin stimulation;but after ATP was added, maturated IL-1β was released into the supernatants of the cells.Moreover, metformin dose-dependently increased the protein levels of both maturated IL-1β and active caspase-1 released by the LPS-activated peritoneal macrophages upon ATP stimulation.Conclusion Metformin intensifies the activation of inflammasome and increases the release of active caspase-1 and maturated IL-1β upon ATP stimulation in the LPS-activated peritoneal macrophages, which should promote inflammatory responses.

Objective: To comparatively study the atrial ifbrillation (AF) in patients with Han, Uygur, Kazak and Hui ethnic groups in Urumqi city.
Methods: A total of 1510 AF patients treated in 12 hospitals in Urumqi city from 2008-01 to 2012-12 were retrospectively studied. There were 1310 patients enrolled in our research including the 4 ethnic groups of Han, n=995 (75.95%), Uygur, n=168 (12.82%), Kazak, n=55 (4.20%) and Hui, n=92 (7.02%).
Results: ①The gender ratios were similar in 4 ethnic groups, P>0.05, while the AF type, cardiac function and risk factors were different, all P<0.05.②The blood pressure was similar in 4 ethnic groups, P>0.05, while the blood routine test, biochemistry and cardiac ultrasound examination were different, all P<0.05.③The treatments were different among 4 ethnic groups, all P<0.05.
Conclusion: The AF patients were different in AF type, biochemistry, cardiac ultrasound and function, anti-coagulation treatment among 4 ethnic groups of Han, Uygur, Kazak and Hui in Urumqi city.