AIM： To generate dendritic cells （DCs） from human peripheral blood and to detect the expression of dendritic cell-specific intercellular adhesion molecule 3 grabbing nonintegrin （DC-SIGN; CD209） for the further s...AIM： To generate dendritic cells （DCs） from human peripheral blood and to detect the expression of dendritic cell-specific intercellular adhesion molecule 3 grabbing nonintegrin （DC-SIGN; CD209） for the further study of DC-SIGN in hepatitis C virus （HCV） transmission. METHODS： Peripheral blood monocytes were isolated from blood of healthy individuals by Ficoll--Hypaque sedimentation and cultured in complete medium containing rhGM-CSF and rhIL-4. Cells were cultured for seven days, with cytokine addition every two days to obtain immature DCs. Characteristics of the cultured cells were observed under light and scanning microscope, and the expression of DC-SIGN was detected by immunofluorescence staining. RESULTS： After seven-day culture, a large number of cells with typical characteristics of DCs appeared. Their characteristics were observed under light and scanning electron microscope. These cells had a variety of cell shapes such as those of bipolar elongate cells, elaborate stellate cells and DCs. DC-SIGN was detected by immunofluorescence staining and its expression level on cultivated dendritic cells was high. CONCLUSION： DCs with a high expression of DC-SIGN can be generated from human peripheral blood monocytes in complete medium containing rhGM-CSF and rhIL-4.展开更多
Objective:To detect the expression of(Peripheral blood mononuclear cells,PBMCs)NLRP3 signal pathway in peripheral blood monocytes of patients with chronic heart failure,and to explore the expression of NLRP3 signal pa...Objective:To detect the expression of(Peripheral blood mononuclear cells,PBMCs)NLRP3 signal pathway in peripheral blood monocytes of patients with chronic heart failure,and to explore the expression of NLRP3 signal pathway and its induced inflammatory response in PBMCs of patients with different types of chronic heart failure.Methods:patients with chronic heart failure(NYHAⅡ~Ⅳ),Ⅱ(nude 20),Ⅲ(nude 20)andⅣ(nude 20)admitted to our hospital from 2019 to 2020 were selected,and 20 normal subjects were selected as the control group.The peripheral venous blood of all subjects was collected,and the plasma and monocytes were extracted respectively.The monocytes were identified by magnetic beads sorting.The mRNA and protein expression levels of NLRP3,ASC and Caspase-1 in PBMCs were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)and Western blot,and the level of interleukin-1β(IL-1β)in plasma was detected by enzyme-linked immunosorbent assay((ELISA)).Results:compared with the normal control group,the expression of NLRP3,ASC,Caspase-1 protein and mRNA in PBMCs of patients with gradeⅡ,ⅢandⅣincreased.Compared with patients with gradeⅡ,the expression of these indexes increased in patients with gradeⅢandⅣ.Compared with patients with gradeⅢ,the expression of these indexes increased in patients with gradeⅣ.Compared with the normal control group,the plasma levels of IL-1βin patients with gradeⅡ,ⅢandⅣwere higher than those in patients with gradeⅡ,ⅢandⅣ(P<0.05).The expression of these indexes in patients with gradeⅢandⅣwas higher than that in patients with gradeⅢ(P<0.05).Conclusion:the results suggest that NLRP3-ASC-Caspase-1 signal pathway may cause chronic inflammation in patients with heart failure and play a role in the progression of chronic heart failure.展开更多
[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated f...[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated from pig, and precursor dendritic cells were obtained by adherence method. The dendritic cells were treated by recombinant porcine granulocyte-monocyte colony stimulating factor （rpGM-CSF） and recombinant porcine interleukin-4 （rplL-4） together, and lipopolysaccharide （LPS） respectively. The cells in different time periods were collected. The morphology of the collected cells was observed by scanning electron microscopy; the expression of surface molecules and phagocytic ability to FITC-dextran were detected by flow cy- tometry; and the stimulating ability for allogeneic T cells was detected by mixed lymphocyte reaction. [Result] The DCs suffering maturation induction in vitro showed typical dendritic morphology; compared with those of DCs untreated by LPS, the cell surface expression of CDla, CD80, CD86, SLAII and CD172a of DCs treated by LPS was significantly increased, the phagocytic ability was reduced slightly, and the stimulating ability for allogeneic T cells was enhanced to some extent. [Conclusion] An in vitro culture method was successfully established for porcine MoDCs in this study, laying a foundation for further study on the role of porcine MoDCs in immunoregulation and anti-virus infection.展开更多
Our recent studies with cultured retinal pigment epithelium cells suggested that overexpression of interleukin 17 receptor C（IL-17RC）,a phenomenon observed in peripheral blood and chorioretinal tissues with age-rela...Our recent studies with cultured retinal pigment epithelium cells suggested that overexpression of interleukin 17 receptor C（IL-17RC）,a phenomenon observed in peripheral blood and chorioretinal tissues with age-related macular degeneration（AMD）,was associated with altered activation of phosphatidylinositide 3-kinase（PI3K）,Akt,and glycogen synthase kinase 3（GSK3）.We wondered whether or not altered PI3 K,Akt,and GSK3 activities could be detected in peripheral blood mononuclear cells（PBMC） obtained from AMD patients.In the patients＇ PBMC,absent or reduced serine-phosphorylation of GSK3α or GSK3β was observed,which was accompanied with increased phosphorylation of GSK3 substrates（e.g.CCAAT enhancer binding protein a,insulin receptor substrate 1,and TAU）,indicative of enhanced GSK3 activation.In addition,decreased protein mass of PI3K85α and tyrosinephosphorylation of PI3K50α was present in PBMC of the AMD patients,suggesting impaired PI3 K activation.Moreover,abnormally lowered molecular weight forms of Akt and GSK3 were detected in PBMC of the AMD patients.These data demonstrate that despite the presence of high levels of IL-17 RC,Wnt-3a and vascular endothelial growth factor,the PI3K/Akt/GSK3 signaling pathway is insensitive to these stimuli in PBMC of the AMD patients.Thus,measurement of PI3K/Akt/GSK3 expression and activity in PBMC may serve as a surrogate biomarker for AMD.展开更多
Day 100 prognostic factors post-autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT) to predict clinical outcomes in diffuse large B-cell lymphoma (DLBCL) patients have not been studied. Thus,...Day 100 prognostic factors post-autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT) to predict clinical outcomes in diffuse large B-cell lymphoma (DLBCL) patients have not been studied. Thus, we retrospectively examined if day 100 absolute monocyte/lymphocyte prognostic score (AMLPS-100) affects clinical outcomes by landmark analysis from day 100 post-APBHSCT in DLBCL. Only DLBCL patients in complete remission at day 100 post-APBHSCT were evaluated. From 2000 to 2007, 134 consecutive DLBCL patients are qualified for the study. Patients with a day 100 absolute monocyte count (AMC-100) ≥ 630 cells/μL and day 100 absolute lymphocyte count (ALC-100) ≤ 1000 cells/μL experienced inferior overall survival (OS) and progression free survival (PFS). On multivariate analysis, the AMC-100 and ALC-100 remained independent predictors of OS and PFS. Combining both values into the AMLPS-100, the 5-year OS rates for low, intermediate, and high AMLPS-100 risk groups were 94% (95% CI, 83.0% - 98.1%), 70% (95% CI, 58.6% - 80.1%), and 13% (95% CI, 3.4% - 40.5%), respectively;and the 5-year PFS rates were 87% (95% CI, 74.0% - 94.1%), 68% (95% CI, 56.0% - 77.8%), and 13% (95% CI, 3.4% - 40.5%), respectively. The AMLPS-100 is a simple biomarker score that can stratify clinical outcomes from day 100 post-APBHSCT in DLBCL patients.展开更多
目的探讨红花多糖（safflower polysaccharide，sPs）体外对人外周血单个核细胞（PBMC）和CD8＋T细胞增殖作用的影响。方法采用葡聚糖-泛影葡胺密度梯度离心法（ficoll-hypaque density gradient centrifugation）从健康成人外周血中分...目的探讨红花多糖（safflower polysaccharide，sPs）体外对人外周血单个核细胞（PBMC）和CD8＋T细胞增殖作用的影响。方法采用葡聚糖-泛影葡胺密度梯度离心法（ficoll-hypaque density gradient centrifugation）从健康成人外周血中分离PBMC，在体外与不同浓度的SPS共同培养，用3H-TdR法检测PBMC增殖活性；流式细胞术检测CI）8＋T细胞的增殖情况。结果SPS能够促进PBMC增殖，尤其1．25g·L-1和0．625g·L-1。两组PBMC增殖作用明显，与对照组比较差异有统计学意义（P〈0．05）；SPS对CD8＋T细胞的增殖有促进作用，与对照组比较差异有统计学意义（P〈0．05）。结论SPS可促进PBMC、CD8＋T细胞的增殖，增强机体非特异性和特异性免疫功能。展开更多
基金Supported by the National Natural Science Foundation of China,No.30170822
文摘AIM： To generate dendritic cells （DCs） from human peripheral blood and to detect the expression of dendritic cell-specific intercellular adhesion molecule 3 grabbing nonintegrin （DC-SIGN; CD209） for the further study of DC-SIGN in hepatitis C virus （HCV） transmission. METHODS： Peripheral blood monocytes were isolated from blood of healthy individuals by Ficoll--Hypaque sedimentation and cultured in complete medium containing rhGM-CSF and rhIL-4. Cells were cultured for seven days, with cytokine addition every two days to obtain immature DCs. Characteristics of the cultured cells were observed under light and scanning microscope, and the expression of DC-SIGN was detected by immunofluorescence staining. RESULTS： After seven-day culture, a large number of cells with typical characteristics of DCs appeared. Their characteristics were observed under light and scanning electron microscope. These cells had a variety of cell shapes such as those of bipolar elongate cells, elaborate stellate cells and DCs. DC-SIGN was detected by immunofluorescence staining and its expression level on cultivated dendritic cells was high. CONCLUSION： DCs with a high expression of DC-SIGN can be generated from human peripheral blood monocytes in complete medium containing rhGM-CSF and rhIL-4.
基金National Natural Science Foundation of China(No.81770297)Anhui Natural Science Foundatio(No.1908085QH353)+1 种基金Anhui Department of Education(No.KJ2018ZD023)Bengbu Medical College Natural Science Foundation Key Project(No.BYKY1833ZD)。
文摘Objective:To detect the expression of(Peripheral blood mononuclear cells,PBMCs)NLRP3 signal pathway in peripheral blood monocytes of patients with chronic heart failure,and to explore the expression of NLRP3 signal pathway and its induced inflammatory response in PBMCs of patients with different types of chronic heart failure.Methods:patients with chronic heart failure(NYHAⅡ~Ⅳ),Ⅱ(nude 20),Ⅲ(nude 20)andⅣ(nude 20)admitted to our hospital from 2019 to 2020 were selected,and 20 normal subjects were selected as the control group.The peripheral venous blood of all subjects was collected,and the plasma and monocytes were extracted respectively.The monocytes were identified by magnetic beads sorting.The mRNA and protein expression levels of NLRP3,ASC and Caspase-1 in PBMCs were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)and Western blot,and the level of interleukin-1β(IL-1β)in plasma was detected by enzyme-linked immunosorbent assay((ELISA)).Results:compared with the normal control group,the expression of NLRP3,ASC,Caspase-1 protein and mRNA in PBMCs of patients with gradeⅡ,ⅢandⅣincreased.Compared with patients with gradeⅡ,the expression of these indexes increased in patients with gradeⅢandⅣ.Compared with patients with gradeⅢ,the expression of these indexes increased in patients with gradeⅣ.Compared with the normal control group,the plasma levels of IL-1βin patients with gradeⅡ,ⅢandⅣwere higher than those in patients with gradeⅡ,ⅢandⅣ(P<0.05).The expression of these indexes in patients with gradeⅢandⅣwas higher than that in patients with gradeⅢ(P<0.05).Conclusion:the results suggest that NLRP3-ASC-Caspase-1 signal pathway may cause chronic inflammation in patients with heart failure and play a role in the progression of chronic heart failure.
基金Supported by Fundamental and Advanced Research Projects of Henan Province(152300410076,2015-2017)Key Science and Technology Program of Henan Province(152102110048,2015-2017)~~
文摘[Objective] This study aimed to establish an in vitro culture model for porcine peripheral blood monocyte-derived dendritic cells （MoDCs）. [Method] Fresh peripheral blood mononuclear cells （PBMCs） were separated from pig, and precursor dendritic cells were obtained by adherence method. The dendritic cells were treated by recombinant porcine granulocyte-monocyte colony stimulating factor （rpGM-CSF） and recombinant porcine interleukin-4 （rplL-4） together, and lipopolysaccharide （LPS） respectively. The cells in different time periods were collected. The morphology of the collected cells was observed by scanning electron microscopy; the expression of surface molecules and phagocytic ability to FITC-dextran were detected by flow cy- tometry; and the stimulating ability for allogeneic T cells was detected by mixed lymphocyte reaction. [Result] The DCs suffering maturation induction in vitro showed typical dendritic morphology; compared with those of DCs untreated by LPS, the cell surface expression of CDla, CD80, CD86, SLAII and CD172a of DCs treated by LPS was significantly increased, the phagocytic ability was reduced slightly, and the stimulating ability for allogeneic T cells was enhanced to some extent. [Conclusion] An in vitro culture method was successfully established for porcine MoDCs in this study, laying a foundation for further study on the role of porcine MoDCs in immunoregulation and anti-virus infection.
基金supported by intramural research funding of National Center for Complementary and Alternative Medicine(now is National Center for Complementary and Integrative Health),NIH,the US Department of Health and Human Services(to X.L.)and an operating grant(MOP 123279)from Canadian Institutes for Health Research(to Z.Y.)
文摘Our recent studies with cultured retinal pigment epithelium cells suggested that overexpression of interleukin 17 receptor C（IL-17RC）,a phenomenon observed in peripheral blood and chorioretinal tissues with age-related macular degeneration（AMD）,was associated with altered activation of phosphatidylinositide 3-kinase（PI3K）,Akt,and glycogen synthase kinase 3（GSK3）.We wondered whether or not altered PI3 K,Akt,and GSK3 activities could be detected in peripheral blood mononuclear cells（PBMC） obtained from AMD patients.In the patients＇ PBMC,absent or reduced serine-phosphorylation of GSK3α or GSK3β was observed,which was accompanied with increased phosphorylation of GSK3 substrates（e.g.CCAAT enhancer binding protein a,insulin receptor substrate 1,and TAU）,indicative of enhanced GSK3 activation.In addition,decreased protein mass of PI3K85α and tyrosinephosphorylation of PI3K50α was present in PBMC of the AMD patients,suggesting impaired PI3 K activation.Moreover,abnormally lowered molecular weight forms of Akt and GSK3 were detected in PBMC of the AMD patients.These data demonstrate that despite the presence of high levels of IL-17 RC,Wnt-3a and vascular endothelial growth factor,the PI3K/Akt/GSK3 signaling pathway is insensitive to these stimuli in PBMC of the AMD patients.Thus,measurement of PI3K/Akt/GSK3 expression and activity in PBMC may serve as a surrogate biomarker for AMD.
文摘Day 100 prognostic factors post-autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT) to predict clinical outcomes in diffuse large B-cell lymphoma (DLBCL) patients have not been studied. Thus, we retrospectively examined if day 100 absolute monocyte/lymphocyte prognostic score (AMLPS-100) affects clinical outcomes by landmark analysis from day 100 post-APBHSCT in DLBCL. Only DLBCL patients in complete remission at day 100 post-APBHSCT were evaluated. From 2000 to 2007, 134 consecutive DLBCL patients are qualified for the study. Patients with a day 100 absolute monocyte count (AMC-100) ≥ 630 cells/μL and day 100 absolute lymphocyte count (ALC-100) ≤ 1000 cells/μL experienced inferior overall survival (OS) and progression free survival (PFS). On multivariate analysis, the AMC-100 and ALC-100 remained independent predictors of OS and PFS. Combining both values into the AMLPS-100, the 5-year OS rates for low, intermediate, and high AMLPS-100 risk groups were 94% (95% CI, 83.0% - 98.1%), 70% (95% CI, 58.6% - 80.1%), and 13% (95% CI, 3.4% - 40.5%), respectively;and the 5-year PFS rates were 87% (95% CI, 74.0% - 94.1%), 68% (95% CI, 56.0% - 77.8%), and 13% (95% CI, 3.4% - 40.5%), respectively. The AMLPS-100 is a simple biomarker score that can stratify clinical outcomes from day 100 post-APBHSCT in DLBCL patients.
文摘目的探讨红花多糖（safflower polysaccharide，sPs）体外对人外周血单个核细胞（PBMC）和CD8＋T细胞增殖作用的影响。方法采用葡聚糖-泛影葡胺密度梯度离心法（ficoll-hypaque density gradient centrifugation）从健康成人外周血中分离PBMC，在体外与不同浓度的SPS共同培养，用3H-TdR法检测PBMC增殖活性；流式细胞术检测CI）8＋T细胞的增殖情况。结果SPS能够促进PBMC增殖，尤其1．25g·L-1和0．625g·L-1。两组PBMC增殖作用明显，与对照组比较差异有统计学意义（P〈0．05）；SPS对CD8＋T细胞的增殖有促进作用，与对照组比较差异有统计学意义（P〈0．05）。结论SPS可促进PBMC、CD8＋T细胞的增殖，增强机体非特异性和特异性免疫功能。
