目的探讨血必净注射液调控重症中暑单核细胞人白细胞DR抗原（HLA-DR）表达异常和血管内皮细胞损伤的效果。方法选取2013年7月~2015年7月在广东省东莞市第五人民医院重症医学科经确诊并住院治疗的重症中暑患者22例为研究对象,采用随机数...目的探讨血必净注射液调控重症中暑单核细胞人白细胞DR抗原（HLA-DR）表达异常和血管内皮细胞损伤的效果。方法选取2013年7月~2015年7月在广东省东莞市第五人民医院重症医学科经确诊并住院治疗的重症中暑患者22例为研究对象,采用随机数字表法分为实验组和对照组,每组各11例。对照组患者给予＂四早一支持＂的西医常规综合治疗,实验组患者在对照组的治疗基础上,加用血必净注射液100 m L静脉滴注,2次/d,连续7 d。治疗前和治疗后第5天,采用流式细胞仪检测循环血中单核细胞HLA-DR表达情况;采用ELISA检测血栓调节蛋白（TM）和血管假性血友病因子（VWF）水平;采用等密度梯度离心法测定循环内皮细胞（CEC）。结果治疗前,两组患者循环血中单核细胞HLA-DR表达较正常人群明显降低（P〈0.05）;治疗后第5天,两组患者循环血中单核细胞HLA-DR表达较治疗前显著升高（P〈0.05）,且实验组较对照组升高明显（P〈0.05）。治疗前,两组患者TM和VWF水平及CEC计数比较,差异无统计学意义（P〉0.05）;治疗后第5天,两组患者TM和VWF水平及CEC计数均较治疗前显著降低（P〈0.05）,且实验组较对照组降低明显（P〈0.05）。结论血必净注射液能有效改善重症中暑患者单核细胞HLA-DR表达异常和血管内皮细胞损伤。展开更多
Objective To investigate the effects of stimulant for nucleotide-binding oligomerization domain 1 （NOD1） on secretion of proinflammatory chemokine/cytokines and insulin-dependent glucose uptake in human differentiat...Objective To investigate the effects of stimulant for nucleotide-binding oligomerization domain 1 （NOD1） on secretion of proinflammatory chemokine/cytokines and insulin-dependent glucose uptake in human differentiated adipocytes. Methods Adipose tissues were obtained from patients undergoing liposuction. Stromal vascular cells were extracted and differentiated into adipocytes. A specific ligand for NOD1, was administered to human adipocytes in culture. Nuclear factor-κB transcriptional activity and proinflammatory chemokine/cytokines production were determined by reporter plasmid assay and enzyme-linked immunosorbent assay, respectively. Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[ 3 H] glucose uptake assay. Furthermore, chemokine/cytokine secretion and glucose uptake in adipocytes transfected with small interfering RNA （siRNA） targeting NOD1 upon stimulation of NOD1 ligand were analyzed. Results Nuclear factor-κB transcriptional activity and monocyte chemoattractant protein-1 （MCP-1）, interleukin （IL）-6, and IL-8 secretion in human adipocytes were markedly increased stimulated with NOD1 ligand （all P〈0.01）. Insulin-induced glucose uptake was decreased upon the activation of NOD1 （P〈0.05）. NOD1 gene silencing by siRNA reduced NOD1 ligand-induced MCP-1, IL-6, and IL-8 release and increased insulin-induced glucose uptake （all P〈0.05）. Conclusion NOD1 activation in adipocytes might be implicated in the onset of insulin resistance.展开更多
Objective. To evaluate the possibility and efficiency of nanoparticle as a new vector in specific gene transference.Methods. Nanoparticle-DNA complex was prepared with Poly- dl-lactic-co-glycolic acid (PLGA) bearing a...Objective. To evaluate the possibility and efficiency of nanoparticle as a new vector in specific gene transference.Methods. Nanoparticle-DNA complex was prepared with Poly- dl-lactic-co-glycolic acid (PLGA) bearing anti-sense monocyte chemotactic protein-1 (A-MCP-l), a specific expression gene, and the package efficiency, release progress in vitro, and the size of the complex were determined. The possibility of the new vector was evaluated with genomic DNA PCR by transferring gene into cultured smooth muscle cells (SMC), cationic lipids as a control. For study in vivo, jugular vein-to-artery bypass grafting procedures were performed on 20 New Zealand white rabbits, of which 6 grafts were transferred with nanoparticle-A-MCP-1 (200 μg), 6 with A - MCP-1 (200 μ g) by cationic liposome, 4 with LNCX plasmid, and 4 as control. Fourteen days after the grafts were harvested, the expression of A-MCP-l and its effect on MCP-1 in vein grafts were detected by dot blot, and the morphologic evaluation of grafts was performed.Results. The package efficiency of the nanoparticle-DNA complex was 0. 9% , release progress in vitro lasted 2 weeks, and the size ranged from 150 to 300nm. SMC genomic DNA PCR showed that A-MCP-l gene could be successfully transfected into cells by nanoparticle. The study in vivo indicated that A-MCP-l mRNA was expressed in both local gene delivery groups, nanoparticle and liposome, meanwhile, MCP-1 expression in vein grafts was significantly inhibited and neointimal hyperplasia was notably reduced.Conclusion. Nanoparticle can act as a vector to transfect specific gene.展开更多
Objective To investigate expression differences of neutrophil and mononuclear phagocyte related gene mRNAs among acute myocardial infarction （AMI）, stable angina （SA） and control groups, and then discuss their exp...Objective To investigate expression differences of neutrophil and mononuclear phagocyte related gene mRNAs among acute myocardial infarction （AMI）, stable angina （SA） and control groups, and then discuss their expression characteristics in the stable angina pectoris （SAP） and AMI stages of coronary artery disease （CAD）. Methods Whole Human Genome Oligo Microarrays were applied to assess the differential expression characteristics of neutrophil and mononuclear phagocyte related mRNAs in patients with AMI （n = 20）, SA （n = 20） and controls （n = 20）. Results （1） Almost all colony-stimulating factors （CSF） and their receptors related mRNAs was up-regulated in AMI and SA groups compared with the control group, and the expression of granulocyte-macrophage colony stimulating factor receptor （GM-CSFR） and granulocyte colony stimulating factor receptor （G-CSFR） mRNAs in the AMI group was significantly up-regulated compared with the other two groups （P 〈 0.01）. （2） The expression of mRNAs related to monocyte chemoattractant protein-1 （MCP-1）, CCR2 （MCP-1 receptor） and CXCR2 （IL-8 receptor） was significantly up-regulated （P 〈 0.01） in AMI group compared with SA and control groups IL-8 mRNA expression in the AMI group was clearly higher than the controls （P 〈 0.05）. （3） All mRNAs expression related to opsonic re- ceptors （IgG FoR and C3bR/C4bR） was significantly up-regulated in AMI group compared with SA and control group （P 〈 0.01）, and the SA group showed an upward trend compared with controls. （4） Most pattern recognition receptor （PRR）-related mRNAs expression was up-regulated in AMI group compared with SA and control groups. Most toll-like receptor （TLR） mRNAs expression was significantly up-regulated （P 〈 0.01） than the SA and control groups, macrophage scavenger receptor （MSR） mRNA was significantly up-regulated in AMI group compared with the control group （P 〈 0.01）, and the SA group showed an upward trend compared with the controls. Conclusions The expression of most neutrophil and mononuclear-macrophage function related genes mRNAs was significantly up-regulated by stages during the progression of CAD, suggesting that the adhesive, chemotactic and phagocytic functions of neutrophil and mononudear-macrophage were strengthened in the occurrence and development of coronary atherosclerosis and AMI. This also showed a stepped up- ward trend as the disease progressed.展开更多
Objective:To investigate the actions of cytokine profile in the immune cells by a seven amino acid peptide mimic from HVR1 of HCV (GQTYTSG,named 7P).Methods:The peripheral blood mononuclear cells (PBMCs) from 17 healt...Objective:To investigate the actions of cytokine profile in the immune cells by a seven amino acid peptide mimic from HVR1 of HCV (GQTYTSG,named 7P).Methods:The peripheral blood mononuclear cells (PBMCs) from 17 healthy blood donors were stimulated with 7P,and the cytokine levels in CD4+CD8-,CD4-CD8+ T cells,NK,NKT cells were analyzed by the intracellular cytokine staining.Results:The frequency of cells which secreted interferon-γ (IFN-γ) was found to be significantly increased in all cells,interleukin-10(IL-10) was significantly increased in CD4+CD8-,CD4-CD8+ T cells but decreased in NK,NKT cells,and tumor necrosis factor-α (TNF-α) was decreased in CD4+CD8-,CD4-CD8+ but increased in NK cells.Conclusion:7P could induce a cytokine profile in different immune cells in vitro and there was some difference between the CD4+CD8-,CD4-CD8+ T cells which represented the adaptive immunity cells and NK,NKT cells which represented the innate immunity cells.This kind of variation of cytokine profile might contribute to anti-virus and anti-inflammatory immune reaction.展开更多
Objective To evaluate the expression and function activity of P-glycoprotein (P-gp) in human mononuclear cells (MNCs) in vitro transfected by multidrug resistance-1(MDR1) mRNA. Methods Two MDR1 cDNA vectors, pT7TS_MDR...Objective To evaluate the expression and function activity of P-glycoprotein (P-gp) in human mononuclear cells (MNCs) in vitro transfected by multidrug resistance-1(MDR1) mRNA. Methods Two MDR1 cDNA vectors, pT7TS_MDR1 and pGEM5Zf(+)_MDR1, were constructed and transcripted in vitro. Vector pGEM5Zf(+)_MDR1 only contained the coding region of mdr1 cDNA, and pT7TS_MDR1 also included Xeponus β-globin 5’ and 3’ untranslated region. MNCs were prepared from peripheral blood of parvicellular lung cancer patient. The two human mdr1 mRNAs were then transferred into human MNCs in vitro by DOTAP. And the expression efficiency and pump function of P-gp were measured with flow cytometry. Results Expression of P-gp significantly elevated in both transferred cells compared with untransferred cells (P < 0.01). And pT7TS_MDR1 showed higher capability in elevating the expression of P-gp than pGEM5Zf(+)_MDR1 (P < 0.01). The P-gp function was elevated in both pT7TS_MDR1 and pGEM5Zf(+)_MDR1 groups. The survival ratio of MNCs in erythrocyte-lysis-solution (ELS, 86.07%) and lymphocyte-isolation-solution (LIS, 83.67%) had no significant difference. The CD34+ cells content of the MNCs used for transfection was 2.65% and 1.01% in ELS and LIS group, respectively (P < 0.01).Conclusions It is a feasible approach to improve P-gp expression in human MNCs by transfection of MDR-1 mRNA. And the ELS may be more suitable for purifing MNCs for mRNA transfection than LIS.展开更多
Objective: To study the chemotactic superfamily genes expression profiling of macrophage line U937 treated with monocyte chemoattractant protein-1 (MCP-1) using gene chip technique. Methods: Total RNA from macrophage ...Objective: To study the chemotactic superfamily genes expression profiling of macrophage line U937 treated with monocyte chemoattractant protein-1 (MCP-1) using gene chip technique. Methods: Total RNA from macrophage line U937 (as control) and U937 with MCP-1 was extracted, made reverse transcript to cDNA and tested with gene expression chip HO2 human. Results: Some chemotactic-related gene expressions were changed in all analyzed genes. Regulated upon activation, normal T cell expressed and secreted (RANTES) was up-regulated over 2-fold and 7 chemotactic-related genes (CCR2, CCR5, CCL16, GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2) were down-regulated over 2-fold in MCP-1 treated U937 cells at mRNA level. Conclusion: MCP-1 can influence some chemokines and receptors expression in macrophage in vitro, in which MCP-1 mainly down-regulates the chemotactic genes expression of those influencing neutrophilic granulocyte (GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2). Another novel finding is that it can also down-regulate the mRNA level of CCR5, which plays a critical role in many disorders and illnesses.展开更多
文摘目的探讨血必净注射液调控重症中暑单核细胞人白细胞DR抗原（HLA-DR）表达异常和血管内皮细胞损伤的效果。方法选取2013年7月~2015年7月在广东省东莞市第五人民医院重症医学科经确诊并住院治疗的重症中暑患者22例为研究对象,采用随机数字表法分为实验组和对照组,每组各11例。对照组患者给予＂四早一支持＂的西医常规综合治疗,实验组患者在对照组的治疗基础上,加用血必净注射液100 m L静脉滴注,2次/d,连续7 d。治疗前和治疗后第5天,采用流式细胞仪检测循环血中单核细胞HLA-DR表达情况;采用ELISA检测血栓调节蛋白（TM）和血管假性血友病因子（VWF）水平;采用等密度梯度离心法测定循环内皮细胞（CEC）。结果治疗前,两组患者循环血中单核细胞HLA-DR表达较正常人群明显降低（P〈0.05）;治疗后第5天,两组患者循环血中单核细胞HLA-DR表达较治疗前显著升高（P〈0.05）,且实验组较对照组升高明显（P〈0.05）。治疗前,两组患者TM和VWF水平及CEC计数比较,差异无统计学意义（P〉0.05）;治疗后第5天,两组患者TM和VWF水平及CEC计数均较治疗前显著降低（P〈0.05）,且实验组较对照组降低明显（P〈0.05）。结论血必净注射液能有效改善重症中暑患者单核细胞HLA-DR表达异常和血管内皮细胞损伤。
基金Supported by Grant from Department of Education of Liaoning Province(2008810)
文摘Objective To investigate the effects of stimulant for nucleotide-binding oligomerization domain 1 （NOD1） on secretion of proinflammatory chemokine/cytokines and insulin-dependent glucose uptake in human differentiated adipocytes. Methods Adipose tissues were obtained from patients undergoing liposuction. Stromal vascular cells were extracted and differentiated into adipocytes. A specific ligand for NOD1, was administered to human adipocytes in culture. Nuclear factor-κB transcriptional activity and proinflammatory chemokine/cytokines production were determined by reporter plasmid assay and enzyme-linked immunosorbent assay, respectively. Insulin-stimulated glucose uptake was measured by 2-deoxy-D-[ 3 H] glucose uptake assay. Furthermore, chemokine/cytokine secretion and glucose uptake in adipocytes transfected with small interfering RNA （siRNA） targeting NOD1 upon stimulation of NOD1 ligand were analyzed. Results Nuclear factor-κB transcriptional activity and monocyte chemoattractant protein-1 （MCP-1）, interleukin （IL）-6, and IL-8 secretion in human adipocytes were markedly increased stimulated with NOD1 ligand （all P〈0.01）. Insulin-induced glucose uptake was decreased upon the activation of NOD1 （P〈0.05）. NOD1 gene silencing by siRNA reduced NOD1 ligand-induced MCP-1, IL-6, and IL-8 release and increased insulin-induced glucose uptake （all P〈0.05）. Conclusion NOD1 activation in adipocytes might be implicated in the onset of insulin resistance.
基金This program was supported by the National Natural Sciences Foundation of China (No. 39870196).
文摘Objective. To evaluate the possibility and efficiency of nanoparticle as a new vector in specific gene transference.Methods. Nanoparticle-DNA complex was prepared with Poly- dl-lactic-co-glycolic acid (PLGA) bearing anti-sense monocyte chemotactic protein-1 (A-MCP-l), a specific expression gene, and the package efficiency, release progress in vitro, and the size of the complex were determined. The possibility of the new vector was evaluated with genomic DNA PCR by transferring gene into cultured smooth muscle cells (SMC), cationic lipids as a control. For study in vivo, jugular vein-to-artery bypass grafting procedures were performed on 20 New Zealand white rabbits, of which 6 grafts were transferred with nanoparticle-A-MCP-1 (200 μg), 6 with A - MCP-1 (200 μ g) by cationic liposome, 4 with LNCX plasmid, and 4 as control. Fourteen days after the grafts were harvested, the expression of A-MCP-l and its effect on MCP-1 in vein grafts were detected by dot blot, and the morphologic evaluation of grafts was performed.Results. The package efficiency of the nanoparticle-DNA complex was 0. 9% , release progress in vitro lasted 2 weeks, and the size ranged from 150 to 300nm. SMC genomic DNA PCR showed that A-MCP-l gene could be successfully transfected into cells by nanoparticle. The study in vivo indicated that A-MCP-l mRNA was expressed in both local gene delivery groups, nanoparticle and liposome, meanwhile, MCP-1 expression in vein grafts was significantly inhibited and neointimal hyperplasia was notably reduced.Conclusion. Nanoparticle can act as a vector to transfect specific gene.
文摘Objective To investigate expression differences of neutrophil and mononuclear phagocyte related gene mRNAs among acute myocardial infarction （AMI）, stable angina （SA） and control groups, and then discuss their expression characteristics in the stable angina pectoris （SAP） and AMI stages of coronary artery disease （CAD）. Methods Whole Human Genome Oligo Microarrays were applied to assess the differential expression characteristics of neutrophil and mononuclear phagocyte related mRNAs in patients with AMI （n = 20）, SA （n = 20） and controls （n = 20）. Results （1） Almost all colony-stimulating factors （CSF） and their receptors related mRNAs was up-regulated in AMI and SA groups compared with the control group, and the expression of granulocyte-macrophage colony stimulating factor receptor （GM-CSFR） and granulocyte colony stimulating factor receptor （G-CSFR） mRNAs in the AMI group was significantly up-regulated compared with the other two groups （P 〈 0.01）. （2） The expression of mRNAs related to monocyte chemoattractant protein-1 （MCP-1）, CCR2 （MCP-1 receptor） and CXCR2 （IL-8 receptor） was significantly up-regulated （P 〈 0.01） in AMI group compared with SA and control groups IL-8 mRNA expression in the AMI group was clearly higher than the controls （P 〈 0.05）. （3） All mRNAs expression related to opsonic re- ceptors （IgG FoR and C3bR/C4bR） was significantly up-regulated in AMI group compared with SA and control group （P 〈 0.01）, and the SA group showed an upward trend compared with controls. （4） Most pattern recognition receptor （PRR）-related mRNAs expression was up-regulated in AMI group compared with SA and control groups. Most toll-like receptor （TLR） mRNAs expression was significantly up-regulated （P 〈 0.01） than the SA and control groups, macrophage scavenger receptor （MSR） mRNA was significantly up-regulated in AMI group compared with the control group （P 〈 0.01）, and the SA group showed an upward trend compared with the controls. Conclusions The expression of most neutrophil and mononuclear-macrophage function related genes mRNAs was significantly up-regulated by stages during the progression of CAD, suggesting that the adhesive, chemotactic and phagocytic functions of neutrophil and mononudear-macrophage were strengthened in the occurrence and development of coronary atherosclerosis and AMI. This also showed a stepped up- ward trend as the disease progressed.
基金Supported by the National High Technology Research and Development Program of China(863 Program,No. 2006AA022494)
文摘Objective:To investigate the actions of cytokine profile in the immune cells by a seven amino acid peptide mimic from HVR1 of HCV (GQTYTSG,named 7P).Methods:The peripheral blood mononuclear cells (PBMCs) from 17 healthy blood donors were stimulated with 7P,and the cytokine levels in CD4+CD8-,CD4-CD8+ T cells,NK,NKT cells were analyzed by the intracellular cytokine staining.Results:The frequency of cells which secreted interferon-γ (IFN-γ) was found to be significantly increased in all cells,interleukin-10(IL-10) was significantly increased in CD4+CD8-,CD4-CD8+ T cells but decreased in NK,NKT cells,and tumor necrosis factor-α (TNF-α) was decreased in CD4+CD8-,CD4-CD8+ but increased in NK cells.Conclusion:7P could induce a cytokine profile in different immune cells in vitro and there was some difference between the CD4+CD8-,CD4-CD8+ T cells which represented the adaptive immunity cells and NK,NKT cells which represented the innate immunity cells.This kind of variation of cytokine profile might contribute to anti-virus and anti-inflammatory immune reaction.
文摘Objective To evaluate the expression and function activity of P-glycoprotein (P-gp) in human mononuclear cells (MNCs) in vitro transfected by multidrug resistance-1(MDR1) mRNA. Methods Two MDR1 cDNA vectors, pT7TS_MDR1 and pGEM5Zf(+)_MDR1, were constructed and transcripted in vitro. Vector pGEM5Zf(+)_MDR1 only contained the coding region of mdr1 cDNA, and pT7TS_MDR1 also included Xeponus β-globin 5’ and 3’ untranslated region. MNCs were prepared from peripheral blood of parvicellular lung cancer patient. The two human mdr1 mRNAs were then transferred into human MNCs in vitro by DOTAP. And the expression efficiency and pump function of P-gp were measured with flow cytometry. Results Expression of P-gp significantly elevated in both transferred cells compared with untransferred cells (P < 0.01). And pT7TS_MDR1 showed higher capability in elevating the expression of P-gp than pGEM5Zf(+)_MDR1 (P < 0.01). The P-gp function was elevated in both pT7TS_MDR1 and pGEM5Zf(+)_MDR1 groups. The survival ratio of MNCs in erythrocyte-lysis-solution (ELS, 86.07%) and lymphocyte-isolation-solution (LIS, 83.67%) had no significant difference. The CD34+ cells content of the MNCs used for transfection was 2.65% and 1.01% in ELS and LIS group, respectively (P < 0.01).Conclusions It is a feasible approach to improve P-gp expression in human MNCs by transfection of MDR-1 mRNA. And the ELS may be more suitable for purifing MNCs for mRNA transfection than LIS.
文摘Objective: To study the chemotactic superfamily genes expression profiling of macrophage line U937 treated with monocyte chemoattractant protein-1 (MCP-1) using gene chip technique. Methods: Total RNA from macrophage line U937 (as control) and U937 with MCP-1 was extracted, made reverse transcript to cDNA and tested with gene expression chip HO2 human. Results: Some chemotactic-related gene expressions were changed in all analyzed genes. Regulated upon activation, normal T cell expressed and secreted (RANTES) was up-regulated over 2-fold and 7 chemotactic-related genes (CCR2, CCR5, CCL16, GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2) were down-regulated over 2-fold in MCP-1 treated U937 cells at mRNA level. Conclusion: MCP-1 can influence some chemokines and receptors expression in macrophage in vitro, in which MCP-1 mainly down-regulates the chemotactic genes expression of those influencing neutrophilic granulocyte (GROβ, GROγ, IL-8 and granulocyte chemotactic protein 2). Another novel finding is that it can also down-regulate the mRNA level of CCR5, which plays a critical role in many disorders and illnesses.
