During the past few years,mounting evidence has proved that the protection induced by vaccines or the prognosis of diseases is correlated with the quantity and quality of T cell responses in both mouse models and human infectious diseases.Examination of one or a few effector functions might not fully reflect the antigen-specific immunity.Recent studies demonstrated that after antigen stimulation,T cells could be induced to produce two or more effector functions simultaneously at single cell level,including cytokines,chemokines production and cytotoxic related molecules,which are named as polyfunctional T cells.Further studies indicated that these polyfunctional T cells are linked to protection from infectious diseases.Therefore,measuring multiple functions of T cells simultaneously is essential for the proper evaluation of specific immunity and may have substantial implications for the design of vaccines.

Objective This study is to study the age related changes of the expression of caspase-3 and the apoptosis states of neurons in primary auditory cortex of 15 young C57BL/6J mice(2 months, 15～20 g) and 15 old C57BL/6J mice(10 months, 50～60 g) and to determine probable physical effects underlying these changes. This paper also discusses the relationship of caspase-3 and apotosis states in primary auditory cortex, the possible role of caspase-3 in primary auditory cortex and the pathogenesis of presbycusis. Methods The immunohistochemical methods were applied to explore the differences of the expression of caspase-3 and the apoptosis states determined by TUNEI. method in the primary auditory cortex between young and old C57BL/6J mice. Results The expression of caspase-3 and apoptosis in Al of old C57BL/6J mice was significantly higher than that in the counterpart of young C57BL/6J mice. Conclusion The results presented a direct morphological evidence for the strengthening of caspase-3 in the primary auditory cortex in old C57BL/6J mice. This method has an important role in the formation and development of presbycusis. The expression of caspase-3 participated in the regulatory procedure of apoptosis and may be a possible factor of the etiologies of presbycusis.

Objective:To investigate the effects of sinomenine(SIN)on the activation and function of T-cells from SLE patients and to elucidate the mechanisms for the immunosuppressive effect of SIN.Methods:PBMCs were isolated from SLE patients and stimulated with anti-CD3 in the presence or absence of different concentrations of SIN.The cytokines were measured by ELISIA,and the activation and function of T-cells were analyzed by flow cytometry.Results:SIN could inhibit IFN-?,IL-2 and IL-4 production of T cells in SLE patients' peripheral blood.In addition,SIN could significantly inhibit the expression of activation molecules CD69 and CD25 by CD4+ and CD8+ T cells.Conclusion:SIN could inhibit immune responses of SLE patients by suppressing T cells activation and cytokine secretion.

Objective To investigate the cellular immune state and pathological mechanism of HBV chronic infection by analyzing the cytokines produced by peripheral blood mononuclear cells(PBMCs) of asymptomatic HBV carriers.Methods PBMCs were prepared from diagnosed chronic asymptomatic HBV individuals and cultured in the presence of different antigens and antibodies.The levels of IFN-?,IL-12,TNF-?,TGF-? and IL-10 in the culture supernatants were respectively detected by ELISA.Results The levels of IFN-?,IL-12 were significantly lower than in the counterparts from healthy controls;meanwhile,those of TGF-? and IL-10 were elevated markedly.Neutralization of TGF-? and IL-10 simultaneously restore the IFN-? production from PBMCs of HBV carriers,and exogenous IL-12 in low dose combined with specific HBV antigens promoted IFN-? production.Conclusion Reduction of IL-12 from PBMCs may be the fundamental reason of viral persistence in HBV chronic carriers,combined IL-12 and specific HBV antigen promoted the celluar immunity of PBMCs from HBV carriers.

Objective:To elucidate the characterization of antigen-specific memory T cells from PPD~+ individuals after stimulation with BCG in vitro.Methods:PBMCs were isolated from PPD~ -/+ normal human peripheral blood and stimulated with BCG. The level of IFN-? in the culture supernatants was assayed by ELISA and the frequency of IFN-?-producinging cells was detected by ELISPOT. The subsets and frequency of cytokine-producing cells were determined at a single cell level by flow cytometry.Results:After stimulation with BCG, PBMCs from PPD~+ but not PPD~- individuals produced significantly high levels of IFN-? in culture supernatants detected by ELISA(P

Objective: IFN-y is produced by both activated T and NK cells in response to mitogen or antigen and has a broad range of immunoregulatory activity. IL-12 has been described as a strong inducer of IFN-?production and promotes the differentiation of naive CD4+T cells toward the Th1 phenotype, priming them for IFN-?production, and consequent induction of cell-mediated immunity. Aim is to know endogenous production of IL,12 from PBMC inducing production of IFN-?in vitro, which is involving in mechanism of T cells to be activated. Methods: Induced IFT-? secretion from human PBMC by stimulated with anti-CD3 , PHA, anti-CD3 plus anti-CD28 and antigen(MLC) Also inhibited IFN-?production by neutralizing antibodies to IL-12 and IL-12R?1 significantly. Results: IFN-?secretion from human activated PBMC is endogenous IL-12dependent, and activated T cells induce the production of IL-12 from APC by a mechanism involving the interaction between CD40L on T cells and CD40 on APC. Conclusion: These results suggest that endogenous IL-12 plays an important role in the normal host defense against infection by a variety of intracellular pathogens and also plays a central role in the genesis of some forms of immunopathology including autoimmune diseases and transplantation rejections.

Objective:To elucidate the phenotypic and functional characteristics of CD3+ CD4+ and CD3+ CD8+ T in pleural fluid of patients with tuberculous pleurisy.Methods:PBMCs and PFCs were isolated. The frequency of CD3+,CD3+ CD4+,CD3+ CD8+ T cells and the ratio of CD4/CD8 in PFCs were analyzed by FACS. The concentration of IFN-?,IL-2 and TNF-? in the pleural fluid were analyzed by ELISA.Production of IFN-?,IL-2 and TNF-? by CD3+ CD4+ and CD3+ CD8+ T cells was detected by FACS following stimulation with BCG.Results:Higher frequencies of CD3+ CD4+ T cells and CD4+ T/CD8+ T ratio were demonstrated in PFCs when compared with PBMCs from normal donors and TB patients.ELISA analysis demonstrated that significantly high levels of IFN-? and TNF-? were detected in pleural fluid.Further analysis by FACS indicated that IL-2,TNF-? and IFN-? were predominantly secreted by CD3+CD4+T cells but not by CD3+ CD8+ T cells.Conclusion:The frequencies of CD3+ and CD3+ CD4+ T cells are increased in PFCs. Large amounts of Th1 cytokines are detected,which are produced by CD3+ CD4+ T cells,and might play important roles against TB infection.

OBJECTIVE To explore the outcomes of CO2 laser assisted stapedotomy with artificial stapes prostheses in the treatment of advanced otosclerosis. METHODS Between January 2010 and January 2014, 15 patients (16 ears) diagnosed as advanced otosclerosis accepted CO2 laser assisted stapedotomy with artificial stapes implantation in our department. The averaged preoperative air conduction threshold of the speech frequency was 70.21 dB HL, the averaged bone conduction threshold was 38.49 dB HL, the averaged air-bone gap (ABG) was 31.72 dB HL. All cases were followed up for more than 6 months after operation. RESULTS All cases accepted auditory follow up after 6 months postoperatively. The speech frequency average air conduction threshold was 43.7 dB HL, the average bone conduction threshold was 28.95 dB HL, the average ABG was 14.75 dB HL. The ABG≤20 dB was achieved in 9 ears (56.3%) and ABG closure (≤10 dB) was achieved in 6 ears (37.5%). No cases appeared intractable vertigo, sensorineural hearing loss, secondary facial paralysis and other serious complications. CONCLUSION CO2 laser assisted technique reduced the probability of serious complications of stapedotomy, most patients with hearing level improved significantly. It's a safe, practical, relatively economical choice for advanced otosclerosis.

Objective To investigate the expression change of RACK1 ,Src and Bcl‐2 in gastric carcinoma tissue and adjacent carcinomatous tissue .Methods Eighty specimens of gastric carcinoma and adjacent carcinomatous tissues in our hospital from Au‐gust 1 ,2011 to February 1 ,2014 were collected .The immunohistochemistry staining and Western blotting methods were adopted to detect the expression of RACK1 ,Src and Bcl‐2 protein in gastric carcinoma and adjacent carcinomatous tissues ,and their correlation was analyzed and performed the statistical analysis by combining with the clinicopathological data .Results The immunohistochem‐istry staining and Western blotting detection displayed that the expression positive rate and expression level of RACK 1 in gastric carcinoma tissue were obviously lower than those in the adjacent carcinomatous tissue ,while the expression positive rate and ex‐pression level of Src and Bcl‐2 in gastric carcinoma tissue were obviously higher than those in the adjacent carcinomatous tissue ,the differences were statistically significant (P<0 .05) .The RACK1 expression in gastric carcinoma tissue was negatively correlated with the Src and Bcl‐2 expression(r= -0 .632 ,-0 .754 ,P<0 .01) ,while Src had no obvious correlation with Bcl‐2 protein(r=0 .217 ,P>0 .05) .Conclusion The expression of RACK1 in gastric carcinoma tissue is significantly decreased ,while the expres‐sions of Src and Bcl‐2 are increased .

BACKGROUND: Hepatocyte/polymer interface with good biocompatibility is the key factor in bioreactor design andconstruction, however, bioreactor used in the clinical practice currently is not an ideal one.OBJECTIVE: To establish human hepatocyte compatible polypropylene interface and to lay a foundation for establishingbioartificial liver reactor with polypropylene hollow fiber.DESIGN, TIME AND SETTING: The comparative observation, cell compatibility experiment was performed betweenFebruary and October 2003 at Shanghai Jiao Tong University, Shanghai, China.MATERIALS: Polypropylene Photochemical graft polymerization modification technique was used to graft hydrophilicacrylamide groups on the surface of polypropylene membrane by chemical bonds to form modified polypropylenemembrane.METHODS: L02 human hepatoeytes were seeded on polypropylene membrane, modified polypropylene membrane andpolystyrene membrane, and polystyrene membrane was used as normal control.MAIN OUTCOME MEASURES: Static water contact angle of polypropylene membrane before and after graftmodification; morphology, adherent rate and proliferation activity of L02 human hepatocytes on different material surfaces.RESULTS: Static water contact angle after polypropylene membrane graft modification was smaller than that before graftmodification (P < 0.05). The adherent rate of L02 human hepatocytes on the surface of modified polypropylene membranewas 0, and the proliferation activity of them, which grew as spherical aggregates, was markedly higher than that of cells onpolystyrene membrane and polypropylene membrane without graft modification.CONCLUSION: Grafting polyacrylamide on the surface of polypropylene can establish good interface of L02 humanhepatocytes/polypropylene and form hepatocyte spherical aggregates through simple static culture.