Background The protective effects against reperfusion injury of cardioprotective drugs have recently been evaluated and found to be inadequate. Guanxinshutong (GXST), a combination of the traditional herb and Mongol...Background The protective effects against reperfusion injury of cardioprotective drugs have recently been evaluated and found to be inadequate. Guanxinshutong (GXST), a combination of the traditional herb and Mongolian medicine, is effective and safe in treating angina pectoris in clinical trials. We assess the cardioprotective effects of GXST against myocardial ischemia and reperfusion (MI/R) injury in rats and explore its possible mechanism. Methods Forty-five male Sprague Dawley rats were randomized into three groups: non-MlfR group (Sham, n = 15), MUR group treated with vehicle (Control, n = 15) and MI/R group treated with GXST (Drug, n = 15). MI/R was induced by ligation of the left anterior descending coronary artery (LAD) for 30 minutes, followed by 2/24 hour reperfusion in the Control and Drug groups. In the Sham group, the LAD was exposed without occlusion. GXST powder (in the Drug group) or saline (in the Control and Sham groups) were administered via direct gastric gavage from 7 day prior to surgery. Blood samples were collected from the carotid artery (10 rats each group) after 2 hours of reperfusion, to determine the levels of tumor necrosis factor-or (TNF-ct), interleukin-1 ~ (IL-113), interleukin-6 (IL-6) and intercellular adhesion molecule-1 (ICAM-1) using enzyme-linked immunosorbent assays. The animals were then sacrificed and the hearts were harvested for histopathology and western blot analysis. Infarct size was measured in the remaining five rats in each group after 24 hours reperfusion. Results GXST significantly decreased levels of TNF-ct, IL-1β, IL-6, ICAM-1, apoptosis index (AI) and infarct size. GXST also obviously inhibited nuclear factor kappa B (NF.r,B) activity when compared with the Control group (all P 〈 0.05). Conclusions GXST is effective in protecting the myocardium against MI/R injury in rats. Its possible cardioprotective mechanism involves inhibition of the inflammatory response and apoptosis following MI/R injury.展开更多
A model describing the absorption process of SO2 into limestone slurry with a spray scrubber is presented. Both the physical performance of the spray liquid in the scrubber and the involved chemical reactions are anal...A model describing the absorption process of SO2 into limestone slurry with a spray scrubber is presented. Both the physical performance of the spray liquid in the scrubber and the involved chemical reactions are analyzed in the model. A con- tinuous concentration change of H+ was solved by iterative coupling using Matlab, and it was found that there was a remarkable influence on the concentration of the other elements in the process of SO2 absorption. The calculations show that the enhancement factor exponentially grows with an increasing value of pH and logarithmically decays with an increasing value of the driving force. To verify the accuracy of the model, experiments were also carried out, and the results suggest that the model, after combining the physical performance of the spray and the enhancement factor, can more precisely describe SO2 absorption in a spray scrubber. Furthermore, a commercial computational fluid dynamics (CFD) tool is used to perform several simulations which describe and clarify the effects of variables on SO2 absorption. The results of numerical simulation can provide a basis for further design and optimization of the scrubber.展开更多
AIM: To elucidate the molecular and cellular features responsible for the increase of regulatory T cells (Tregs) in gastric cancer. METHODS: The frequencies of CD4 + Foxp3 + Tregs and the level of transforming growth ...AIM: To elucidate the molecular and cellular features responsible for the increase of regulatory T cells (Tregs) in gastric cancer. METHODS: The frequencies of CD4 + Foxp3 + Tregs and the level of transforming growth factor-β1 (TGF-β1) were analyzed from 56 patients with gastric cancer byflow cytometry and enzyme-linked immunosorbent assay respectively. Foxp3 gene expression was analyzed by real-time polymerase chain reaction. The gastric cancer microenvironment was modeled by establishing the coculture of gastric cancer cell line, MGC-803, with sorting CD4 + T cells. The normal gastric mucosa cell line, GES-1, was used as the control. The production of TGF-β1 was detected in supernatant of MGC and GES-1. The carboxyfluorescein diacetatesuccinimidyl ester (CFSE) dilution assay was performed to evaluate the proliferation characteristics of induced Tregs. Neutralizing anti-TGF-β1 antibody was added to the co-culture system for neutralization experiments. RESULTS: The level of serum TGF-β1 in gastric cancer patients (15.1 ± 5.5 ng/mL) was significantly higher than that of the genderand age-matched healthy controls (10.3 ± 3.4 ng/mL) (P < 0.05). Furthermore, the higher TGF-β1 level correlated with the increased population of CD4 + Foxp3 + Tregs in advanced gastric cancer (r = 0.576, P < 0.05). A significant higher frequency of CD4 + Foxp3 + Tregs was observed in PBMCs cultured with the supernatant of MGC than GES-1 (10.6% ± 0.6% vs 8.7% ± 0.7%, P < 0.05). Moreover, using the purified CD4 + CD25 T cells, we confirmed that the increased Tregs were mainly induced from the conversation of CD4 + CD25 naive T cells, and induced Tregs were functional and able to suppress the proliferation of effector T cells. Finally, we demonstrated that gastric cancer cells induced the increased CD4 + Foxp3 + Tregs via producing TGF-β1. Gastric cancer cells upregulated the production of TGF-β1 and blockade of TGF-β1 partly abrogated Tregs phenotype. CONCLUSION: Gastric cancer cell can induce Tregs development via producing TGF-β1, by which the existence of cross-talk between the tumor and immune cells might regulate anti-tumor immune responses.展开更多
AIM:To investigate the effect of emodin on pancreatic claudin-5 and occludin expression,and pancreatic paracellular permeability in acute pancreatitis(AP).METHODS:Experimental pancreatitis was induced by retrograde in...AIM:To investigate the effect of emodin on pancreatic claudin-5 and occludin expression,and pancreatic paracellular permeability in acute pancreatitis(AP).METHODS:Experimental pancreatitis was induced by retrograde injection of 5% sodium taurocholate into the biliopancreatic duct.Emodin was injected via the external jugular vein 0 or 6 h after induction of AP.Rats from sham operation and AP groups were injected with normal saline at the same time.Samples of pancreas were obtained 6 or 12 h after drug administration.Pancreatic morphology was examined with hematoxylin and eosin staining.Pancreatic edema was estimated by measuring tissue water content.Tumor necrosis factor(TNF)-α and interleukin(IL)-6 level were measured by enzyme-linked immunosorbent assay.Pancreatic paracellular permeability was assessed by tissue dye extravasation.Expression of pancreatic claudin-5 and occludin was examined by immunohistology,quantitative real-time reverse transcriptase polymerase chain reaction and western blotting.RESULTS:Pancreatic TNF-α and IL-6 levels,wet/dry ratio,dye extravasation,and histological score were significantly elevated at 3,6 and 12 h following sodium taurocholate infusion;treatment with emodin prevented these changes at all time points.Immunostaining of claudin-5 and occludin was detected in rat pancreas,which was distributed in pancreatic acinar cells,ductal cells and vascular endothelial cells,respectively.Sodium taurocholate infusion significantly decreased pancreatic claudin-5 and occludin mRNA and protein levels at 3,6 and 12 h,and that could be promoted by intravenous administration of emodin at all time points.CONCLUSION:These results demonstrate that emodin could promote pancreatic claudin-5 and occludin expression,and reduce pancreatic paracellular permeability.展开更多
Objective To investigate the role of TNF receptor-associated factor 2 (TRAF-2) and TRAF6 in CD40-induced nuclear factor-κB (NF-κB) signaling pathway and whether CD40 signaling requires TRAF2. Methods Human B cell li...Objective To investigate the role of TNF receptor-associated factor 2 (TRAF-2) and TRAF6 in CD40-induced nuclear factor-κB (NF-κB) signaling pathway and whether CD40 signaling requires TRAF2. Methods Human B cell lines were transfected with plasmids expressing wild type TRAF2 or dominant negative TRAF2,TRAF2-shRNA,or TRAF6-shRNA. The activation of NF-κB was detected by Western blot,kinase assay,transfactor enzyme-linked immunosorbent assay (ELISA),and fluorescence resonance energy transfer (FRET). Analysis of the role of TRAF-2 and TRAF-6 in CD40-mediated NF-κB activity was examined following stimulation with recombinant CD154. Results TRAF2 induced activity of IκB-kinases (IKKα,IKKi/ε),phosphorylation of IκBα,as well as nuclear translocation and phosphorylation of p65/RelA. In contrast,TRAF6 strongly induced NF-κB activation and nuclear translocation of p65 as well as p50 and c-Rel. Engagement of CD154-induced nuclear translocation of p65 was inhibited by a TRAF6-shRNA,but conversely was enhanced by a TRAF2-shRNA. Examination of direct interactions between CD40 and TRAFs by FRET documented that both TRAF2 and TRAF6 directly interacted with CD40. However,the two TRAFs competed for CD40 binding. Conclusions These results indicate that TRAF2 can signal in human B cells,but it is not essential for CD40-mediated NF-κB activation. Moreover,TRAF2 can compete with TRAF6 for CD40 binding,and thereby limit the capacity of CD40 engagement to induce NF-κB activation.展开更多
AIM: To investigate genetic diversity of Helicobacter pylori (H. pylorl) cell division-related gene A (cdrA) and its effect on the host response.METHODS: Inactivation of H. py/ori cdrA, which is involved in ceil...AIM: To investigate genetic diversity of Helicobacter pylori (H. pylorl) cell division-related gene A (cdrA) and its effect on the host response.METHODS: Inactivation of H. py/ori cdrA, which is involved in ceil division and morphological elonga- tion, has a role in chronic persistent infections. Ge- netic property of H. pylori cdrA was evaluated using polymerase chain reaction and sequencing in 128 (77 American and 51 Japanese) clinical isolates obtained from 48 and 51 patients, respectively. Enzyme-linked immunosorbent assay was performed to measure in- terleukin-8 (IL-8) secretion with gastric biopsy speci- mens obtained from American patients colonized with cdrA-positive or -negative strains and AGS cells co- cultured with wild-type HPK5 (cdrA-positive) or its de- rivative HPKT510 (cdrA-disruptant). Furthermore, the cytotoxin-associated gene A (cagA) status (transloca- tion and phosphorylation) and kinetics of transcription factors [nuclear factor-kappa B (NF-~:B) and inhibition kappa B] were investigated in AGS cells co-cultured with HPK5, HPKT510 and its derivative HPKSCA (cagA- disruptant) by western blotting analysis with immuno- precipitation. RESULTS: Genetic diversity of the H. pylori cdrA gene demonstrated that the cdrA status segregated into two categories including four allele types, cdrA-positive (al- lele types, I and 11 ) and cdrA-negative (allele types; 111 and IV) categories, respectively. Almost all Japanese isolates were cdrA-positive ( 1 : 7.8% and 11 : 90.2%), whereas 16.9% of American isolates were cdrA-positive (11) and 83.1% were cdrA-negative (nl: 37.7% and IV: 45.5%), indicating extended diversity of cdrA in individual American isolates. Comparison of each isolate from different regions (antrum and corpus) in the stomach of 29 Americans revealed that cdrA status was identical in both isolates from different regions in 17 cases. However, 12 cases had a different cdrA al- lele and 6 of them exhibited a different cdrA category between two regions in the stomach. Furthermore, in 5 of the 6 cases possessing a different cdrA category, cdrA-negative isolate existed in the corpus, suggesting that cdrA-negative strain is more adaptable to coloni- zation in the corpus. IL-8 secretions from AGS revealed that IL-8 levels induced by a cdrA-disrupted HPKT510 was significantly lower (P 〈 0.01) compared to wild- type HPK5: corresponding to 50%-60% of those of wild-type HPK5. These data coincided with in vivo data that an average value of IL-8 in biopsy specimens from cdrA-positive and cdrA-negative groups was 215.6 and 135.9 pg/mL, respectively. Western blotting analysis documented that HPKT510 had no effect on CagA translocation and phosphorylation, however, nuclear accumulation of NF-κB was lower by HPKT510 com- pared to HPK5. CONCLUSION: Colonization by a cdrA-negative or cdrA-dysfunctional strain resulted in decreased IL-8 production and repression of NF-κB, and hence, atten- uate the host immunity leading to persistent infection.展开更多
Objective:The aim of our study was to investigate the immune status of patients with rectal cancer and its relationship with clinicopathological features.Methods:The serum levels of interleukin-8(IL-8),tumor necrosis ...Objective:The aim of our study was to investigate the immune status of patients with rectal cancer and its relationship with clinicopathological features.Methods:The serum levels of interleukin-8(IL-8),tumor necrosis factor(TNF-α) and T-cell subgroup contents were measured using a double-antibody sandwich assay of ELISA in 43 patients with rectal cancer,and compared with the normal health adults.Results:In patients with rectal cancer,the serum levels of CD4,CD4/CD8 of T-cell subgroup in peripheral blood were significantly lower than the control group(P < 0.01),which gradually decreased with increase of Dukes stage;but the levels of CD8,IL-8 and TNF-α were higher than the control group,which gradually increased with increase of Dukes stage.Conclusion:The immunocompromice exists in patients with rectal cancer,there is a correlation between the contents of T-cell subgroup,IL-8 and TNF-α in serum and the Dukes stage of rectal cancer.Therefore immunotherapy can be used in patients with rectal cancer.展开更多
Tryptophan residues of IgGRF from rheumatoid arthritis patient were modified with N-Bromosuccinimide (NBS) and the product of modification was characterized by UV-spectrum、Fluorescence emission spectrum and CD-spectr...Tryptophan residues of IgGRF from rheumatoid arthritis patient were modified with N-Bromosuccinimide (NBS) and the product of modification was characterized by UV-spectrum、Fluorescence emission spectrum and CD-spectrum. The corresponding adsorption capacity of immobilized ssDNA immunoadsorbent for IgGRF was enhanced from 46% to 86%, This result indicates the tryptophan residue is essential for the interaction between ssDNA and IgGRF .and ionic-bonding plays an important role in adsorption.展开更多
Objective: We studied the inhibitory effect of human cholangiocarcinoma line (QBC939) treated by selective cyclooxygenase-2 (COX-2) inhibitor NS-398 and provided the theoretical foundation for the clinical practi...Objective: We studied the inhibitory effect of human cholangiocarcinoma line (QBC939) treated by selective cyclooxygenase-2 (COX-2) inhibitor NS-398 and provided the theoretical foundation for the clinical practice. Methods: Selec- tive COX-2 inhibitor NS-398 on the growth suppression was evaluated by MTT method. The apoptotic rate was quantified and cell cycle by flow cytometry (FCM). Invasive ability was detected by Transwell. The expression of vascular endothelial growth factor (VEGF) and COX-2 in cholangiocarcinoma line was determined by enzyme-linked immunosorbent assay (ELISA). Results: NS-398 could be time-dose dependently inhibited the growth, induced apoptosis, invasived ability, S phase was inhibited, down-regulate the expression of VEGF and COX-2 in cholangiocarcinoma line (QBC939). Conclusion: NS398 can inhibit proliferation of cholangiocarcinoma cell line QBC939 and inhibit the invasive ability and induce its apoptosis in vitro, which may contributed to the COX-2 dependent pathway to reduce the release of VEGF.展开更多
Objective: The aim of our study was to analyze intedeukin-18 (IL-18) and vascular endothelial growth factor (VEGF) serum levels in patients with prostate cancer before and after operation and the possible correla...Objective: The aim of our study was to analyze intedeukin-18 (IL-18) and vascular endothelial growth factor (VEGF) serum levels in patients with prostate cancer before and after operation and the possible correlation between IL-18 and VEGF serum levels. Methods: Serum IL-18 and VEGF levels were measured by enzyme-linked immunosorbent assay (ELISA) in 36 patients with prostate cancer before and after radical prostatectomy and in 25 healthy controls. Results: Serum IL-18 and VEGF levels were significantly higher in patients with prostate cancer before operation with respect to healthy controls (P 〈 0.05), with a highly significant correlation between IL-18 and VEGF (R = 0.800, P = 0.017). It was significantly reduced in IL-18 and VEGF after operation. IL-18 and VEGF serum concentrations were correlated with the clinicalopathologi- cal status of patients with prostate cancer. Conclusion: It is correlative with serum IL-18 and VEGF. Serum IL-18 and VEGF levels may be useful prognostic marker in patients with prostate cancer.展开更多
OBJECTIVE: To observe effect of Liuweibuqi Capsule, a Traditional Chinese Medicine (TCM), on the janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway and matrix metalloproteinases ...OBJECTIVE: To observe effect of Liuweibuqi Capsule, a Traditional Chinese Medicine (TCM), on the janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway and matrix metalloproteinases (MMPs) in a chronic obstructive pulmonary disease (COPD) rat model with lung deficiency in terms of TCM's pattern differentiation. METHODS: Rats were randomly divided into a normal group, model group, Liuweibuqi group, Jinshuibao group, and spleen aminopeptidase group (n= 10). Aside from the normal group, all rats were ex-posed to smoke plus lipopolysaccharide tracheal instillation to establish the COPD model with lung deficiency. Models were established after 28 days and then the normal and model groups were given normal saline (0.09 g/kg), Liuweibuqi group was given Liuweibuqi capsule (0.35 g/kg), Jinshuibao group was given Jinshuibao capsules (0.495 g/kg), and the spleen group was given spleen aminopeptidase (0.33 mg/kg), once a day for 30 days. Changes in symptoms, signs, and lung histology were observed. Lung function was measured with a spirometer. Serum cytokines were detected using enzyme-linked immunosorbent assay, and changes in the JAK/STAT pathway, MMP-9, and MMPs inhibitor 1 (TIMP1) were detected by immunohistochemistry, RT-PCR, and western blotting, respectively.RESULTS: Compared with the normal group, lung tissue was damaged, and lung function was reduced in the model control group. Additionally, the levels of interleukin (IL)-1β, y interferon (IFN-γ), and IL-6 were higher, while IL-4 and IL-10 were lower in the model control group than those in the normal group. The expressions of JAK1, STAT3, ρ-STAT3, and MMP-9 mRNA and protein in lung tissue were higher, and TIMP1 mRNA and protein was lower in the model group compared with the normal group. After treatment, compared with the model group, the expression of inflammatory cytokines was lower in each treatment group, and expressions of JAK/ STAT pathway, MMPs were lower. Compared with the positive control groups, the Jinshuibao and spleen aminopeptidase groups, lung function was better, and JAK1, STAT3, and p-STAT3 protein were lower and TIMP1 was higher in the Liuweibuqi group.CONCLUSION: Liuweibuqi capsules can improve the symptoms of COPD possibly by regulating the expression of the JAK1/STAT3 pathway and MMP9/ TIMP1.展开更多
Protein microarrays based on fluorescence detection have been widely utilized for high-throughput functional proteomic analysis. However, a drawback of such assays has been low sensitivity and narrow dynamic range, li...Protein microarrays based on fluorescence detection have been widely utilized for high-throughput functional proteomic analysis. However, a drawback of such assays has been low sensitivity and narrow dynamic range, limiting their capabilities, especially for detecting low abundance biological molecules such as cytokines in human samples. Here, we present fluorescence-enhancing microarrays on plasmonic gold films for multiplexed cytokine detection with up to three orders of magnitude higher sensitivity than on conventional nitrocellulose and glass substrates. Cytokine detection on the gold plasmonic substrate is about one to two orders of magnitude more sensitive than enzyme-linked immunosorbent assay (ELISA) and can be multiplexed. A panel of six cytokines (Vascular endothelial growth factor (VEGF), Interleukin 1β (IL-1β), Interleukin 4 (IL-4), Interleukin 6 (IL-6), Interferon γ (IFN-γ), and Tumor necrosis factor (TNF)) were detected in the culture media of cancer cells. This work establishes a new method of high throughput multiplexed cytokine detection with higher sensitivity and dynamic range than ELISA.展开更多
Objective: To explore the feasibility of using regenerated silk fibroin membrane to construct artificial skin substitutes for wound healing, it is necessary to evaluate its cytocompatibility. Methods: The effects of...Objective: To explore the feasibility of using regenerated silk fibroin membrane to construct artificial skin substitutes for wound healing, it is necessary to evaluate its cytocompatibility. Methods: The effects of regenerated silk fibroin film on cytotoxicity, adhesion, cell cycle, and apoptosis of L929 cells, growth and vascular endothelial growth factor (VEGF) expression of ECV304 cells, and VEGF, angiopoietin-1 (Ang-1), platelet-derived growth factor (PDGF) and fibroblast growth factor 2 (FGF2) expression of WI-38 cells were assessed by 3-(4,5)-dimethylthiahiazo (-z-yl)-3,5-di-phenytetrazoliumromide (MTT) assay, viable cell counting, flow cytometry (FCM), and enzyme-linked immunosorbant assay (ELISA). Results: We showed that the regenerated silk fibroin film was not cytotoxic to L929 cells and had no adverse influence on their adhesion, cell cycle or apoptosis; it had no adverse influence on the growth and VEGF secretion of ECV304 cells and no effect on the secretion of VEGF, Ang-1, PDGF and FGF2 by WI-38 cells. Conclusion: The regenerated silk fibroin film should be an excellent biomaterial with good cytocompatibility, providing a framework for reparation after trauma in clinical applications.展开更多
文摘Background The protective effects against reperfusion injury of cardioprotective drugs have recently been evaluated and found to be inadequate. Guanxinshutong (GXST), a combination of the traditional herb and Mongolian medicine, is effective and safe in treating angina pectoris in clinical trials. We assess the cardioprotective effects of GXST against myocardial ischemia and reperfusion (MI/R) injury in rats and explore its possible mechanism. Methods Forty-five male Sprague Dawley rats were randomized into three groups: non-MlfR group (Sham, n = 15), MUR group treated with vehicle (Control, n = 15) and MI/R group treated with GXST (Drug, n = 15). MI/R was induced by ligation of the left anterior descending coronary artery (LAD) for 30 minutes, followed by 2/24 hour reperfusion in the Control and Drug groups. In the Sham group, the LAD was exposed without occlusion. GXST powder (in the Drug group) or saline (in the Control and Sham groups) were administered via direct gastric gavage from 7 day prior to surgery. Blood samples were collected from the carotid artery (10 rats each group) after 2 hours of reperfusion, to determine the levels of tumor necrosis factor-or (TNF-ct), interleukin-1 ~ (IL-113), interleukin-6 (IL-6) and intercellular adhesion molecule-1 (ICAM-1) using enzyme-linked immunosorbent assays. The animals were then sacrificed and the hearts were harvested for histopathology and western blot analysis. Infarct size was measured in the remaining five rats in each group after 24 hours reperfusion. Results GXST significantly decreased levels of TNF-ct, IL-1β, IL-6, ICAM-1, apoptosis index (AI) and infarct size. GXST also obviously inhibited nuclear factor kappa B (NF.r,B) activity when compared with the Control group (all P 〈 0.05). Conclusions GXST is effective in protecting the myocardium against MI/R injury in rats. Its possible cardioprotective mechanism involves inhibition of the inflammatory response and apoptosis following MI/R injury.
基金Project supported by the National Key Technologies Supporting Program of China during the 11th Five-Year Plan Period (No.2006BAA01B04)the New Century Excellent Talent in University(No. NCET-06-0513), China
文摘A model describing the absorption process of SO2 into limestone slurry with a spray scrubber is presented. Both the physical performance of the spray liquid in the scrubber and the involved chemical reactions are analyzed in the model. A con- tinuous concentration change of H+ was solved by iterative coupling using Matlab, and it was found that there was a remarkable influence on the concentration of the other elements in the process of SO2 absorption. The calculations show that the enhancement factor exponentially grows with an increasing value of pH and logarithmically decays with an increasing value of the driving force. To verify the accuracy of the model, experiments were also carried out, and the results suggest that the model, after combining the physical performance of the spray and the enhancement factor, can more precisely describe SO2 absorption in a spray scrubber. Furthermore, a commercial computational fluid dynamics (CFD) tool is used to perform several simulations which describe and clarify the effects of variables on SO2 absorption. The results of numerical simulation can provide a basis for further design and optimization of the scrubber.
基金Supported by Shanghai Municipal Natural Science Foundation, No. 10ZR1420000National Natural Science Foundation of China, No. 81072009
文摘AIM: To elucidate the molecular and cellular features responsible for the increase of regulatory T cells (Tregs) in gastric cancer. METHODS: The frequencies of CD4 + Foxp3 + Tregs and the level of transforming growth factor-β1 (TGF-β1) were analyzed from 56 patients with gastric cancer byflow cytometry and enzyme-linked immunosorbent assay respectively. Foxp3 gene expression was analyzed by real-time polymerase chain reaction. The gastric cancer microenvironment was modeled by establishing the coculture of gastric cancer cell line, MGC-803, with sorting CD4 + T cells. The normal gastric mucosa cell line, GES-1, was used as the control. The production of TGF-β1 was detected in supernatant of MGC and GES-1. The carboxyfluorescein diacetatesuccinimidyl ester (CFSE) dilution assay was performed to evaluate the proliferation characteristics of induced Tregs. Neutralizing anti-TGF-β1 antibody was added to the co-culture system for neutralization experiments. RESULTS: The level of serum TGF-β1 in gastric cancer patients (15.1 ± 5.5 ng/mL) was significantly higher than that of the genderand age-matched healthy controls (10.3 ± 3.4 ng/mL) (P < 0.05). Furthermore, the higher TGF-β1 level correlated with the increased population of CD4 + Foxp3 + Tregs in advanced gastric cancer (r = 0.576, P < 0.05). A significant higher frequency of CD4 + Foxp3 + Tregs was observed in PBMCs cultured with the supernatant of MGC than GES-1 (10.6% ± 0.6% vs 8.7% ± 0.7%, P < 0.05). Moreover, using the purified CD4 + CD25 T cells, we confirmed that the increased Tregs were mainly induced from the conversation of CD4 + CD25 naive T cells, and induced Tregs were functional and able to suppress the proliferation of effector T cells. Finally, we demonstrated that gastric cancer cells induced the increased CD4 + Foxp3 + Tregs via producing TGF-β1. Gastric cancer cells upregulated the production of TGF-β1 and blockade of TGF-β1 partly abrogated Tregs phenotype. CONCLUSION: Gastric cancer cell can induce Tregs development via producing TGF-β1, by which the existence of cross-talk between the tumor and immune cells might regulate anti-tumor immune responses.
基金Supported by National Natural Science Foundation of China,No.30500688
文摘AIM:To investigate the effect of emodin on pancreatic claudin-5 and occludin expression,and pancreatic paracellular permeability in acute pancreatitis(AP).METHODS:Experimental pancreatitis was induced by retrograde injection of 5% sodium taurocholate into the biliopancreatic duct.Emodin was injected via the external jugular vein 0 or 6 h after induction of AP.Rats from sham operation and AP groups were injected with normal saline at the same time.Samples of pancreas were obtained 6 or 12 h after drug administration.Pancreatic morphology was examined with hematoxylin and eosin staining.Pancreatic edema was estimated by measuring tissue water content.Tumor necrosis factor(TNF)-α and interleukin(IL)-6 level were measured by enzyme-linked immunosorbent assay.Pancreatic paracellular permeability was assessed by tissue dye extravasation.Expression of pancreatic claudin-5 and occludin was examined by immunohistology,quantitative real-time reverse transcriptase polymerase chain reaction and western blotting.RESULTS:Pancreatic TNF-α and IL-6 levels,wet/dry ratio,dye extravasation,and histological score were significantly elevated at 3,6 and 12 h following sodium taurocholate infusion;treatment with emodin prevented these changes at all time points.Immunostaining of claudin-5 and occludin was detected in rat pancreas,which was distributed in pancreatic acinar cells,ductal cells and vascular endothelial cells,respectively.Sodium taurocholate infusion significantly decreased pancreatic claudin-5 and occludin mRNA and protein levels at 3,6 and 12 h,and that could be promoted by intravenous administration of emodin at all time points.CONCLUSION:These results demonstrate that emodin could promote pancreatic claudin-5 and occludin expression,and reduce pancreatic paracellular permeability.
基金Supported by Key Projects of the National Science & Technology Pillar Program in the Eleventh Five-year Plan Period (2008-BAI59B02)
文摘Objective To investigate the role of TNF receptor-associated factor 2 (TRAF-2) and TRAF6 in CD40-induced nuclear factor-κB (NF-κB) signaling pathway and whether CD40 signaling requires TRAF2. Methods Human B cell lines were transfected with plasmids expressing wild type TRAF2 or dominant negative TRAF2,TRAF2-shRNA,or TRAF6-shRNA. The activation of NF-κB was detected by Western blot,kinase assay,transfactor enzyme-linked immunosorbent assay (ELISA),and fluorescence resonance energy transfer (FRET). Analysis of the role of TRAF-2 and TRAF-6 in CD40-mediated NF-κB activity was examined following stimulation with recombinant CD154. Results TRAF2 induced activity of IκB-kinases (IKKα,IKKi/ε),phosphorylation of IκBα,as well as nuclear translocation and phosphorylation of p65/RelA. In contrast,TRAF6 strongly induced NF-κB activation and nuclear translocation of p65 as well as p50 and c-Rel. Engagement of CD154-induced nuclear translocation of p65 was inhibited by a TRAF6-shRNA,but conversely was enhanced by a TRAF2-shRNA. Examination of direct interactions between CD40 and TRAFs by FRET documented that both TRAF2 and TRAF6 directly interacted with CD40. However,the two TRAFs competed for CD40 binding. Conclusions These results indicate that TRAF2 can signal in human B cells,but it is not essential for CD40-mediated NF-κB activation. Moreover,TRAF2 can compete with TRAF6 for CD40 binding,and thereby limit the capacity of CD40 engagement to induce NF-κB activation.
基金Supported by The Project Research Fund from Kochi University,to Takeuchi Ha Grant-in-Aid for Scientific Research from the Ministry of Education,Science and Culture of Japan,No. 21590631 and 21590629,in part
文摘AIM: To investigate genetic diversity of Helicobacter pylori (H. pylorl) cell division-related gene A (cdrA) and its effect on the host response.METHODS: Inactivation of H. py/ori cdrA, which is involved in ceil division and morphological elonga- tion, has a role in chronic persistent infections. Ge- netic property of H. pylori cdrA was evaluated using polymerase chain reaction and sequencing in 128 (77 American and 51 Japanese) clinical isolates obtained from 48 and 51 patients, respectively. Enzyme-linked immunosorbent assay was performed to measure in- terleukin-8 (IL-8) secretion with gastric biopsy speci- mens obtained from American patients colonized with cdrA-positive or -negative strains and AGS cells co- cultured with wild-type HPK5 (cdrA-positive) or its de- rivative HPKT510 (cdrA-disruptant). Furthermore, the cytotoxin-associated gene A (cagA) status (transloca- tion and phosphorylation) and kinetics of transcription factors [nuclear factor-kappa B (NF-~:B) and inhibition kappa B] were investigated in AGS cells co-cultured with HPK5, HPKT510 and its derivative HPKSCA (cagA- disruptant) by western blotting analysis with immuno- precipitation. RESULTS: Genetic diversity of the H. pylori cdrA gene demonstrated that the cdrA status segregated into two categories including four allele types, cdrA-positive (al- lele types, I and 11 ) and cdrA-negative (allele types; 111 and IV) categories, respectively. Almost all Japanese isolates were cdrA-positive ( 1 : 7.8% and 11 : 90.2%), whereas 16.9% of American isolates were cdrA-positive (11) and 83.1% were cdrA-negative (nl: 37.7% and IV: 45.5%), indicating extended diversity of cdrA in individual American isolates. Comparison of each isolate from different regions (antrum and corpus) in the stomach of 29 Americans revealed that cdrA status was identical in both isolates from different regions in 17 cases. However, 12 cases had a different cdrA al- lele and 6 of them exhibited a different cdrA category between two regions in the stomach. Furthermore, in 5 of the 6 cases possessing a different cdrA category, cdrA-negative isolate existed in the corpus, suggesting that cdrA-negative strain is more adaptable to coloni- zation in the corpus. IL-8 secretions from AGS revealed that IL-8 levels induced by a cdrA-disrupted HPKT510 was significantly lower (P 〈 0.01) compared to wild- type HPK5: corresponding to 50%-60% of those of wild-type HPK5. These data coincided with in vivo data that an average value of IL-8 in biopsy specimens from cdrA-positive and cdrA-negative groups was 215.6 and 135.9 pg/mL, respectively. Western blotting analysis documented that HPKT510 had no effect on CagA translocation and phosphorylation, however, nuclear accumulation of NF-κB was lower by HPKT510 com- pared to HPK5. CONCLUSION: Colonization by a cdrA-negative or cdrA-dysfunctional strain resulted in decreased IL-8 production and repression of NF-κB, and hence, atten- uate the host immunity leading to persistent infection.
文摘Objective:The aim of our study was to investigate the immune status of patients with rectal cancer and its relationship with clinicopathological features.Methods:The serum levels of interleukin-8(IL-8),tumor necrosis factor(TNF-α) and T-cell subgroup contents were measured using a double-antibody sandwich assay of ELISA in 43 patients with rectal cancer,and compared with the normal health adults.Results:In patients with rectal cancer,the serum levels of CD4,CD4/CD8 of T-cell subgroup in peripheral blood were significantly lower than the control group(P < 0.01),which gradually decreased with increase of Dukes stage;but the levels of CD8,IL-8 and TNF-α were higher than the control group,which gradually increased with increase of Dukes stage.Conclusion:The immunocompromice exists in patients with rectal cancer,there is a correlation between the contents of T-cell subgroup,IL-8 and TNF-α in serum and the Dukes stage of rectal cancer.Therefore immunotherapy can be used in patients with rectal cancer.
基金National Basic Science and Development of China (G1999O64707)
文摘Tryptophan residues of IgGRF from rheumatoid arthritis patient were modified with N-Bromosuccinimide (NBS) and the product of modification was characterized by UV-spectrum、Fluorescence emission spectrum and CD-spectrum. The corresponding adsorption capacity of immobilized ssDNA immunoadsorbent for IgGRF was enhanced from 46% to 86%, This result indicates the tryptophan residue is essential for the interaction between ssDNA and IgGRF .and ionic-bonding plays an important role in adsorption.
文摘Objective: We studied the inhibitory effect of human cholangiocarcinoma line (QBC939) treated by selective cyclooxygenase-2 (COX-2) inhibitor NS-398 and provided the theoretical foundation for the clinical practice. Methods: Selec- tive COX-2 inhibitor NS-398 on the growth suppression was evaluated by MTT method. The apoptotic rate was quantified and cell cycle by flow cytometry (FCM). Invasive ability was detected by Transwell. The expression of vascular endothelial growth factor (VEGF) and COX-2 in cholangiocarcinoma line was determined by enzyme-linked immunosorbent assay (ELISA). Results: NS-398 could be time-dose dependently inhibited the growth, induced apoptosis, invasived ability, S phase was inhibited, down-regulate the expression of VEGF and COX-2 in cholangiocarcinoma line (QBC939). Conclusion: NS398 can inhibit proliferation of cholangiocarcinoma cell line QBC939 and inhibit the invasive ability and induce its apoptosis in vitro, which may contributed to the COX-2 dependent pathway to reduce the release of VEGF.
文摘Objective: The aim of our study was to analyze intedeukin-18 (IL-18) and vascular endothelial growth factor (VEGF) serum levels in patients with prostate cancer before and after operation and the possible correlation between IL-18 and VEGF serum levels. Methods: Serum IL-18 and VEGF levels were measured by enzyme-linked immunosorbent assay (ELISA) in 36 patients with prostate cancer before and after radical prostatectomy and in 25 healthy controls. Results: Serum IL-18 and VEGF levels were significantly higher in patients with prostate cancer before operation with respect to healthy controls (P 〈 0.05), with a highly significant correlation between IL-18 and VEGF (R = 0.800, P = 0.017). It was significantly reduced in IL-18 and VEGF after operation. IL-18 and VEGF serum concentrations were correlated with the clinicalopathologi- cal status of patients with prostate cancer. Conclusion: It is correlative with serum IL-18 and VEGF. Serum IL-18 and VEGF levels may be useful prognostic marker in patients with prostate cancer.
基金Supported by The National Natural Science Foundation Project:Study on the Metabolism of Chronic Obstructive Pulmonary Disease Pulmonary Qi Deficiency Syndrome and Cerebral Cortex Correlation Spectrum(No.81072781)
文摘OBJECTIVE: To observe effect of Liuweibuqi Capsule, a Traditional Chinese Medicine (TCM), on the janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway and matrix metalloproteinases (MMPs) in a chronic obstructive pulmonary disease (COPD) rat model with lung deficiency in terms of TCM's pattern differentiation. METHODS: Rats were randomly divided into a normal group, model group, Liuweibuqi group, Jinshuibao group, and spleen aminopeptidase group (n= 10). Aside from the normal group, all rats were ex-posed to smoke plus lipopolysaccharide tracheal instillation to establish the COPD model with lung deficiency. Models were established after 28 days and then the normal and model groups were given normal saline (0.09 g/kg), Liuweibuqi group was given Liuweibuqi capsule (0.35 g/kg), Jinshuibao group was given Jinshuibao capsules (0.495 g/kg), and the spleen group was given spleen aminopeptidase (0.33 mg/kg), once a day for 30 days. Changes in symptoms, signs, and lung histology were observed. Lung function was measured with a spirometer. Serum cytokines were detected using enzyme-linked immunosorbent assay, and changes in the JAK/STAT pathway, MMP-9, and MMPs inhibitor 1 (TIMP1) were detected by immunohistochemistry, RT-PCR, and western blotting, respectively.RESULTS: Compared with the normal group, lung tissue was damaged, and lung function was reduced in the model control group. Additionally, the levels of interleukin (IL)-1β, y interferon (IFN-γ), and IL-6 were higher, while IL-4 and IL-10 were lower in the model control group than those in the normal group. The expressions of JAK1, STAT3, ρ-STAT3, and MMP-9 mRNA and protein in lung tissue were higher, and TIMP1 mRNA and protein was lower in the model group compared with the normal group. After treatment, compared with the model group, the expression of inflammatory cytokines was lower in each treatment group, and expressions of JAK/ STAT pathway, MMPs were lower. Compared with the positive control groups, the Jinshuibao and spleen aminopeptidase groups, lung function was better, and JAK1, STAT3, and p-STAT3 protein were lower and TIMP1 was higher in the Liuweibuqi group.CONCLUSION: Liuweibuqi capsules can improve the symptoms of COPD possibly by regulating the expression of the JAK1/STAT3 pathway and MMP9/ TIMP1.
文摘Protein microarrays based on fluorescence detection have been widely utilized for high-throughput functional proteomic analysis. However, a drawback of such assays has been low sensitivity and narrow dynamic range, limiting their capabilities, especially for detecting low abundance biological molecules such as cytokines in human samples. Here, we present fluorescence-enhancing microarrays on plasmonic gold films for multiplexed cytokine detection with up to three orders of magnitude higher sensitivity than on conventional nitrocellulose and glass substrates. Cytokine detection on the gold plasmonic substrate is about one to two orders of magnitude more sensitive than enzyme-linked immunosorbent assay (ELISA) and can be multiplexed. A panel of six cytokines (Vascular endothelial growth factor (VEGF), Interleukin 1β (IL-1β), Interleukin 4 (IL-4), Interleukin 6 (IL-6), Interferon γ (IFN-γ), and Tumor necrosis factor (TNF)) were detected in the culture media of cancer cells. This work establishes a new method of high throughput multiplexed cytokine detection with higher sensitivity and dynamic range than ELISA.
基金supported by the National Basic Research Program (973) of China (No.2005CB623906)the Medical Development Foundation of Soochow University (No.EE134702),China
文摘Objective: To explore the feasibility of using regenerated silk fibroin membrane to construct artificial skin substitutes for wound healing, it is necessary to evaluate its cytocompatibility. Methods: The effects of regenerated silk fibroin film on cytotoxicity, adhesion, cell cycle, and apoptosis of L929 cells, growth and vascular endothelial growth factor (VEGF) expression of ECV304 cells, and VEGF, angiopoietin-1 (Ang-1), platelet-derived growth factor (PDGF) and fibroblast growth factor 2 (FGF2) expression of WI-38 cells were assessed by 3-(4,5)-dimethylthiahiazo (-z-yl)-3,5-di-phenytetrazoliumromide (MTT) assay, viable cell counting, flow cytometry (FCM), and enzyme-linked immunosorbant assay (ELISA). Results: We showed that the regenerated silk fibroin film was not cytotoxic to L929 cells and had no adverse influence on their adhesion, cell cycle or apoptosis; it had no adverse influence on the growth and VEGF secretion of ECV304 cells and no effect on the secretion of VEGF, Ang-1, PDGF and FGF2 by WI-38 cells. Conclusion: The regenerated silk fibroin film should be an excellent biomaterial with good cytocompatibility, providing a framework for reparation after trauma in clinical applications.