Objective To investigate the effect of tumor associated glycoprotein-72 （TAG-72） redirected T lymphocytes on breast cancer cells. Methods Peripheral blood mononuclear cells （PBMCs）were isolated from healthy volunteers. The recombinant vector anti-TAG-72-scFv-CD3ζ-pcDNA 3.0 were transfected into PBMCs by lipofectamineTM2000 （transfection group）， PBMCs transfected with plasmid pcDNA 3.0 as control group. MCF-7 and Bcap37 cells were cocultivated with PBMCs of transfection group and control group， respectively， and antitumor response of G1 block was observed. Results G1 block rate of MCF-7 cells in transfection group was （82.3±6.9）%， which was significantly higher than that in control group 〔（43.4±3.9）%， P＜0.05〕. G1 block rate of Bcap37 cells in transfection group was （51.3±4.7）%， and not differed from that in control group 〔（45.6±2.5）%， P＞0.05〕. Conclusion TAG-72 redirected T lymphocytes can inhibit the cell proliferation of TAG-72 positive breast cancer cells， and it may provide valuable tools for the cellular immunotherapy.
Objective To determine the application values of gene chip technique in cardiovascular surgical clinical and research work. Microarray for gene expression profiles was used to screen out the differentially expressed genes during cardiopulmonary bypass(CPB) in peripheral blood mononuclear cell. By doing these, it was hoped that some clues in inflammatory response during CPB could be found out. Methods The patients’ oxygenated bloods were drawn immediately before onset and termination of CPB. Peripheral blood mononuclear cell (PBMC) were obtained from heparinised blood by Ficoll gradient centrifugation. The differentially expression was measured using BD AtlasTM cDNA Expression Arrays. The candidate genes were corroborated by semiquantitative reverse transcriptionpolymerase chain reaction (RT-PCR). Results Gene chip technique was successfully used in CPB study. The gene expression profiles of cytokines of PBMC during CPB were screened out. Interleukin 6 and Wnt5a were the differentially expressed genes. But the validity using semiquantitative RT-PCR found no statistically difference(P=0.888,0.135). Conclusion Microarray technique has positive application values in the study of cytokines during CPB. cDNA microarray for gene expression profiles can primarily screen out differentially expression genes during CPB. These genes may be engaged in inflammation and other pathophysiological reactions during CPB. PBMC is not the major source of cytokines during CPB.
【摘要】 目的 检测B细胞成熟抗原（BCMA）mRNA在系统性红斑狼疮（SLE）患者外周血单个核细胞（PBMC）的表达水平，探讨BCMA在SLE发病中的意义。 方法 纳入2006年1-11月收治的36例SLE患者，同期17例健康志愿者作为对照组，采用半定量RT-PCR法检测外周血单个核细胞中BCMA mRNA的表达，并与SLE疾病活动指数（SLEDAI）进行相关性分析。 结果 SLE患者组BCMA mRNA表达水平（0.598±0.230）均明显高于正常对照组（0.411±0.309）（Plt;0.05）。SLE患者BCMA mRNA表达水平与SLEDAI评分无相关性（P=0.590）。 结论 SLE患者BCMA mRNA表达水平的增高，可能在SLE的发病机制中具有一定的作用。【Abstract】 Objective To detect the mRNA expression of B-cell maturation antigen (BCMA) in peripheral blood mononuclear cells (PBMC) in patients with systemic lupus erythematosus (SLE), and explore the role of BCMA in the pathogenesis of SLE. Methods From January 2006 to November 2006 the expression of BCMA mRNA in PBMC of 36 patients with SLE and 17 normal controls were measured by half-quantitative RT-PCR. The linear correlation between the expression of BCMA mRNA and SLE disease activity index (SLEDAI) was assessed. Results The level of BCMA mRNA (0.598±0.230) in PBMC significantly increased in SLE patients compared with that in the normal controls (0.411±0.309) (Plt;0.05). The expression of BCMA mRNA in SLE patients showed no correlation with SLEDAI score (P=0.590). Conclusion The results suggest that the expression of BCMA mRNA might play an important role in the pathogenesis of SLE.
Objective To study the expression of NLRP3 inflammasome and its downstream inflammatory factors in patients with chronic obstructive pulmonary disease (COPD) and healthy controls, and to reveal the effect and significance of NLRP3 inflammasome in the pathogenesis of COPD. Methods Forty patients with acute exacerbation COPD (AECOPD) who were hospitalized from November 2016 to May 2017 were recruited in the AECOPD group, and recruited in the stable COPD group when they entered the stable stage. Forty healthy individuals were recruited in the control group. General information and peripheral blood were collected from each subject. The levels of NLRP3 mRNA and caspase-1 mRNA in peripheral blood mononuclear cells were measured by real-time PCR. The levels of IL-18 and IL-1β were measured by enzyme-linked immunosorbent assay. Results The levels of NLRP3 mRNA, IL-18 and IL-1β in the AECOPD patients were significantly higher than those in the stable COPD group [2.11±0.77, 12.79 (7.10, 43.13) pg/ml, 17.02 (8.36, 52.21) pg/ml vs. 1.60±0.44, 10.66 (6.32, 18.59) pg/ml, 13.34 (7.07, 16.89) pg/ml, all P<0.05] . The levels of NLRP3 mRNA, IL-18 and IL-1β in the AECOPD patients were significantly higher than those in the control group [2.11±0.77, 12.79 (7.10, 43.13) pg/ml, 17.02 (8.36, 52.21) pg/mlvs. 1.00±0.49, 6.29 (4.73, 7.93) pg/ml, 5.93 (4.81, 9.67) pg/ml, all P<0.05]. The levels of NLRP3 mRNA, IL-18 and IL-1β were significantly higher in the stable COPD group than the control group [1.60±0.44, 10.66 (6.32, 18.59) pg/ml, 13.34 (7.07, 16.89) pg/mlvs. (1.00±0.49, 6.29 (4.73, 7.93) pg/ml, 5.93 (4.81, 9.67) pg/ml, all P<0.05]. Correlation analysis showed that the plasma IL-18 level was positive correlated with leukocyte count and neutrophil percentage in the AECOPD group (r=0.372, P<0.05;r=0.386, P<0.05). The expression of NLRP3 mRNA in the AECOPD group and stable COPD group were positively correlated with the CAT score (r=0.387, P<0.05;r=0.399, P<0.05) . Conclusion NLRP3 inflammasome is involved in the inflammatory response in COPD patients.
ObjectiveTo investigate activation of phosphatidylinositol 3 hydroxykinase (PI3K)/AKT pathway and sphingosine 1-phosphate receptor 2 (S1PR2) in peripheral blood mononuclear cells (PBMCs) of acute obstructive cholangitis (AOC) rats and their effects on systemic inflammation in rats.Methods① In vitro experiment: The isolated PBMCs from the rats were divided into 4 groups: a control group, LY294002 treatment group, lipopolysaccharide (LPS) treatment group, and LPS+LY294002 treatment group. The levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the supernatant were detected and the phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the PBMCs were detected. ② In vivo experiment: The rats were randomly divided into four groups: a control group, LY294002 treatment group, AOC model group, and AOC+LY294002 treatment group. The survival rate of rats was recorded, the liver function (ALT, AST, and TBIL), TNF-α, and IL-6 levels in the serum were detected. The phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the PBMCs of the rats were detected. Results① The results of in vitro experiment: The levels of TNF-α and IL-6 in the LPS+LY294002 treatment group were significantly lower than those in the LPS treatment group (P<0.050). The phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the LPS+LY294002 treatment group were significantly lower than those in the LPS treatment group (P<0.050). ② The results of in vivo experiment: The survival rate of rats in the AOC+LY294002 treatment group was higher than those in the AOC group. The serum levels of ALT, AST, TBIL, TNF-α, and IL-6 in the AOC+LY294002 treatment group were significantly lower than those in the AOC model group (P<0.050). The phosphorylation levels of PI3K and AKT and protein level of S1PR2 in the AOC+LY294002 treatment group were significantly lower than those in the AOC model group (P<0.050).ConclusionInhibition of activation of PI3K/AKT pathway in PBMCs can inhibit expression of S1PR2, then alleviate systemic inflammatory response induced by AOC in rats.