Objective To investigate the T cell cytotoxicity induced by recombinant adenovirus carrying HIV-1 vpr gene.Methods C8166 cells infected with rAd-vpr or negative control rAd-vector,were analyzed for cell cycle distribution and cell death by flow cytometry.The discrimination of living cells,apoptotic and necrotic cells were differentiated with Hoechst-PI double staining under the confocal microscopy.Changes of mitochondrial membrane potential(△ψm)were monitored by JC-1 staining method.Results Annexin V-PI and Hoechst-PI staining indicated the death effects of HIV-1 Vpr on C8166 cells.PI flow cytometric analysis showed that cell cycle arrested in G2 phase.C8166 cell△ψm collapse mediated by Vpr was detected by JC-1 fluorescent staining.Conclusion The ability of recombinant adenovirus carrying HIV-1 vpr gene to induce mitochondria dysfunction,cell cycle G2 phase arrest and cell death was confirmed in C8166 cells.

Objective To develop a automatic infusion alarm device. Methods The automatic infusion alarm device was subtly reformed according to the principal of the character of the spring that the different gravity caused its length to be different. It consisted of two parts: springs and alarm circuit controller box. Results Clinical test was undertaken. Conclusions The automatic infusion alarm device is simply manufactured, reliable and cheap. It is valuable to advocate clinically.

AIM:To analyze the effects of oxymatrine(OMT) on the quantity of murine regulatory T cells(Tr cells) in the peripheral blood and mouse lymphocyte proliferation stimulated by Con A,and to probe into the immunological mechanism that OMT treats allergic contact dermatitis(ACD).METHODS:An ACD mouse model stimulated by dinitrofluorobenzene(DNFB) was established.Different dosages of OMT,PBS and hydrocortisone(HCT) were intraperitoneally injected(IP) into the mice.Blood samples were collected at 1 d,7 d,14 d,21 d and 28 d,then the T cells were isolated and marked with anti-CD3,anti-CD4,anti-CD25 three-colored immune fluorescence antibody to detect the quantity of CD4+CD25+ T cells with flow cytometry.The fluorescence intensity changes of lymphocytes which were isolated from mouse's lymph node and co-stimulated by polyclonal stimulator Con A and OMT were examined by carboxyfluorescein diacetate succinimidyl ester(CFDA-SE) staining and flow cytometry.RESULTS:OMT at concentrations of 500,125 and 31 mg/L had the ability to restrain the proliferation of lymphocytes from lymph node in a dose dependent manner.However,OMT at concentrations of 16,8,4 and 2 mg/L promoted the proliferation of T lymphocytes from lymph node,but was not obviously dependent on its concentration.Intraperitoneal injection of OMT increased the numbers of CD4+CD25+T cell in peripheral blood obviously(P

AIM:To study the changes of mitochondria during apoptosis in Jurkat cells induced by arsenic oxide(As2O3).METHODS:By treated with 4?10-6 mol/L As2O3,apoptosis and necrosis of Jurkat cells were assessed by annexin V-FITC/PI double staining flowcytometry.Mitochondrial mass and its membrane potential(△?m)was measured by NAO/PI and DiOC6(3)/PI staining,respectively.Free radical formation was detected by DCFDA staining.RESULTS:After 48 h of As2O3 treatment,the rates of early apoptotic Jurkat cells in As2O3 and control groups were(18.98?1.40)% and(5.17?0.80)%,respectively(P

AIM: To study mitochondrial mass and structural protein changes in dexamethasone (DEX)-mediated mouse thymocyte apoptosis process. METHODS: DEX-induced mouse thymocyte apoptosis model was established. Annexin V-FITC/PI double staining was used to identify apoptotic and necrotic cells by flowcytometry, JC-1 staining was adopted to test mitochondrial membrane potential (△?_m), and cellular structural protein changes were studied with CFDA-SE staining. RESULTS: By 1?10~(-6) mol/L DEX stimulation, the apoptotic rate was 51.25%?5.51% and had significantly difference from control group (12.03%?2.00%); the necrotic rate in DEX group was 30.25%?3.67% and also had significantly difference from control group (10.11%?1.11%, P

AIM: To investigate the relationship between CD200~+CK7~+ trophoblasts and the resorption of embryos in a poly (I∶C)-induced abortion model. METHODS: The status of CD200 expression was investigated in Balb/c?C57BL/6 and Balb/c?Balb/c mice as induced model of embryo-resorption by an i.p. injection of poly (I∶C). CD200 expression on CK7~+ cells from placentas was detected with flow cytometry. CD200~+ cells in placenta were observed with immunocytochemical staining. RESULTS: Both the percentage and absolute number of CD200~+CK7~+ cells were dramatically decreased by injection of poly (I∶C) in Balb/c?C57BL/6 (6.3%?6.2% vs 36.1%?9.3%, P

AIM: To study the effect of natural killer(NK)-cells on graft-versus-host disease(GVHD), graft rejection, engraftment and reconstituting of hematopoiesis in mouse allogeneic bone marrow transplantation. METHODS: Lethally irradiated BALB/c(H-2 d) mice were transplanted with C57/6j(H-2 b) bone marrow containing donor peripheral T cells and/or NK cells. Recipients CD 34 + cell counts and the expression of H-2K b+ cell were detected by flow cytometry, peripheral white blood cell(WBC) was detected by auto-cytometry, and the recipients survival rates, GVHD, engraftment and hematopoiesis recovery were then observed. RESULTS: In the group of transplanted with NK cells infusion, the incidence of GVHD was evidently lessened, the survival rates, WBC and CD 34 + cell counts and the expression of H-2K b+ cell were significantly high than that without NK cells infusion. CONCLUSION: In mouse allogeneic bone marrow transplantation, alloreactivity NK cells prevent GVHD, reduce graft rejection, and promote engraftment and reconstituting of hematopoiesis. [

AIM: To study the effects of [8-(diethylamino) octyl-3, 4, 5 -trimethoxybenzoate] (TMB-8), an intracellular Ca2+ antagonist, on the activation, proliferation and cell-cycle distribution of the mouse T lymphocytes stimulated by concanvalin A (Con A) in vitro. METHODS: After stimulated with Con A, T cells were treated with different concentrations of TMB-8 alone and its combination with cyclosporine A (CsA). The expression of CD69, the early marker of CD3+ T cell activation, was measured by FACS. The proliferation-related index was determined by carboxyl fluorescin diacetate succinmidyl ester (CFDA-SE) flow cytometry. The cell-cycle distribution was analyzed by propidium iodide staining.RESULTS: After 6 h culture, the activation rate of CD69+ T cell in Con A group was (74.88?1.88)%. 10, 20 and 40 ?mol/L of TMB-8 inhibited the expression of CD69 (P

AIM: To investigate the effect of enhanced green fluorescence protein (EGFP) gene transfection on the cell cycle distribution of primary cultured human chondrocytes in order to establish a tracking method of cultured human nasoseptal chondrocytes. METHODS: pEGFP-N1 plasmid was amplified in E.coli, and purified by high purity kit. Primary cultured human chondrocytes,which were initially obtained from the nasoseptal cartilage, were cultured in vitro and transferred with pEGFP-N1 by means of electroporation with Amaxa nucleofector device. Transfering process and transient expression were evaluated by laser scanning confocal microscope (LSCM), the transfer efficiency and the cell cycle distribution were evaluated by flow cytometry. RESULTS: There was significant expression of EGFP at 24 h after transferring. The transfection efficiency of pEGFP-N1 into primary cultured human chondrocytes reached 35 37% at 48 h. It didn't affect the process of cell adherance and had no effect on the cell cycle distribution. CONCLUSION: Primary cultured human chondrocytes, which were transfected with pEGFP, are alive in vitro, and the transferring process doesn't affect the cell cycle distribution. These results suggest that pEGFP-N1 is an ideal transient expression vector for primary cultured human chondrocytes and it might be a well tracer in construction tissue engineered cartilage.

0.05) compared with control group at 1 h and 3 h; while ~FL 1 in DEX group at 5 h (660.91?72.95) was significant lower (P