AIM: To determine the NF-kB activity in peripheral blood mononuclear cells (PBMC) in patients with acute cholangitis of severe type (ACST) and correlate the degree of NF-kB activation with severity of biliary tract in...AIM: To determine the NF-kB activity in peripheral blood mononuclear cells (PBMC) in patients with acute cholangitis of severe type (ACST) and correlate the degree of NF-kB activation with severity of biliary tract infection and clinical outcome. METHODS: Twenty patients with ACST were divided into survivor group (13 cases) and nonsurvivor group (7 cases). Other ten patients undergoing elective gastrectomy or inguinal hernia repair were selected as control group. Peripheral blood samples were taken 24 hours postoperatively. PBMC were separated by density gradient centrifugation, then nuclear proteins were isolated from PBMC, and Electrophoretic Mobility Shift Assay (EMSA) used determined. The results were quantified by scanning densitometer of a Bio-Image Analysis System and expressed as relative optical density (ROD). The levels of TNF-alpha, IL-6, and IL-10 in the plasma of patients with ACST and healthy control subjects were determined by using an enzyme-linked immunoassay (ELISA). RESULTS: The NF-kB activity was 5.02 +/- 1.03 in nonsurvivor group, 2.98 +/- 0.51 in survivor group and 1.06 +/- 0.34 in control group. There were statistical differences in three groups (P【0.05). The levels of TNF-alpha and IL-6 in plasma were (498 +/- 53)ng.L(-1)and (587 +/- 64)ng.L(-1)in nonsurvivor group, (284 +/- 32)ng.L(-1) and (318 +/- 49)ng.L(-1)in survivor group and (89 +/- 11)ng.L(-1) and (102 +/-13)ng.L(-1)in control group. All patients with ACST had increased levels of TNF-alpha and IL-6, which were many-fold greater than those of control group, and there was an evidence of significantly higher levels in those of nonsurvivor group than that in survivor group (P【0.05). The levels of IL-10 in plasma were (378+/-32)ng.L(-1), (384+/-37)ng.L(-1) and (68+/-11)ng.L(-1) in three groups, respectively. All patients had also increased levels of IL-10 when compared with control group (P【0.05), but the IL-10 levels were not significantly higher in nonsurvivors than in survivors (P】0.05). CONCLUSION: NF-kB activity in PBMC in patients with ACST increases markedly and the degree of NF-kB activation is correlated with severity of biliary tract infection and clinical outcome.展开更多
AIM： To evaluate peripheral blood lymphocyte subsets in patients with acute pancreatitis （AP）. METHODS： Twenty patients with mild AP （M-AP） and 15 with severe AP （SAP） were included in our study. Peripheral bl...AIM： To evaluate peripheral blood lymphocyte subsets in patients with acute pancreatitis （AP）. METHODS： Twenty patients with mild AP （M-AP） and 15 with severe AP （SAP） were included in our study. Peripheral blood lymphocytes were examined at d 1-3, 5, 10 and 30 by means of flow cytometry. RESULTS： A significant depletion of circulating lymphocytes was found in AP. In the early AP, the magnitude of depletion was similar for T- and B- lymphocytes. In the late course of S-AP, B-lymphocytes were much more depleted than T-lymphocytes. At d 10, strong shift in the CD7＋/CD19＋ ratio implicating predominance of T- over B-lymphocytes in S-AP was found. Among T-lymphocytes, the significant depletion of the CD4＋ population was observed in M-AP and S-AP, while CDS＋ cells were in the normal range. Lymphocytes were found to strongly express activation markers： CD69, CD25, CD28, CD38 and CD122. Serum interleukin-2 （IL-2）, IL-4, IL-5, IL-10, interferon-γ （IFN-γ） and tumor necrosis factor-α （TNF-α） levels were significantly increased in both forms of AP. The magnitude of elevation of cytokines known to be produced by Th2 was much higher than cytokines produced by Th1 cells. CONCLUSION： AP in humans is characterized by significant reduction of peripheral blood T- and B-lymphocytes.展开更多
AIM： To investigate the role of inflammatory and anti-vira genes in the pathogenesis of SARS. METHODS： cDNA microarrays were used to screen the gene expression profiles of peripheral blood mononuclear cells （PBMCs...AIM： To investigate the role of inflammatory and anti-vira genes in the pathogenesis of SARS. METHODS： cDNA microarrays were used to screen the gene expression profiles of peripheral blood mononuclear cells （PBMCs） in two SARS patients （one in the acute severe phase and the other in the convalescent phase） and a healthy donor. In addition, real-time qualitative PCR was also performed to verify the reproducibility of the microarray results. The data were further analyzed. RESULTS： Many inflammatory and anti-viral genes were differentially expressed in SARS patients. Compared to the healthy control or the convalescent case, plenty of pro-inflammatory cytokines such as IL-1, TNF-α, IL-8, and MAPK signaling pathway were significantly upregulated in the acute severe case. However, anti-inflammatory agents such as IL-4 receptor, IL-13 receptor, IL-1Ra, and TNF-α-induced proteins 3 and 6 also increased dramatically in the acute severe case. On the contrary, a lot of IFN-stimulated genes like PKR, GBP-1 and 2, CXCL-10 and 11, and JAK/STAT signal pathway were downregulated in the acute severe case compared to the convalescent case. CONCLUSION： Gene expression in SAPS patients mirrors a host state of inflammation and anti-viral immunity at the transcription level, and understanding of gene expression profiles may make contribution to further studies of the SAPS pathogenesis.展开更多
AIM:To study the effects of synthetic nonmethylated CpG-containing oligodeoxynucleotides(CpG-ODNs) ,either alone or combined with recombinant Hepatitis B surface antigen(HBsAg) polypeptide,on the phenotype,function,an...AIM:To study the effects of synthetic nonmethylated CpG-containing oligodeoxynucleotides(CpG-ODNs) ,either alone or combined with recombinant Hepatitis B surface antigen(HBsAg) polypeptide,on the phenotype,function,and intracellular signaling pathways of monocyte-derived dendritic cells(DCs) in patients with chronic hepatitis B(CHB) .METHODS:Peripheral blood monocytes isolated from CHB patients and healthy volunteers were induced to be dendritic cells by recombinant human granulocyte-monocyte colony stimulating factor and interleukin-4.The DCs were then treated with CpG-ODNs,CpGODNs/HBsAg,or tumor necrosis factor(TNF)-αfor 18 h.The expression of surface molecules including HLA-DR,CD86,and CD1a in DCs were detected by flow cytometry,and the expression of signal transducers and activators of transcription(STAT1,3,4,5,6) and suppressors of cell signaling(SOCS1,3) were determined by Western blotting assay.In addition,the capacity of DCs to stimulate allogeneic T lymphocytes and the amount of IL-12p70 released from DCs were measured.RESULTS:In the DCs derived from patients with CHB,treatment with TNF-α,CpG-ODNs,or CpG-ODNs/HBsAg,as compared to the vector control,significantly increased the expression of HLA-DR,stimulated the release of IL-12p70,and enhanced the capacity of DCs to stimulate allogenic T lymphocytes.The expressions of STAT1/4/6 and SOCS1/3,but not STAT3/5,were upregulated by TNF-α,CpG-ODNs,and CpG-ODNs/HBsAg.In addition,the expression of CD1a was upregulated only in the presence of both CpG-ODNs and HBsAg.CONCLUSION:The treatment with CpG-ODNs,either alone or combined with HBsAg,has a remarkable stimulatory effect on the impaired phenotype and function of DCs in CHB,possibly by regulating the expression of STAT1,4,6 and SOCS1,3.展开更多
Objective： To assess the effects of penehyclidine hydrochloride on patients with acute lung injury （ALI）, to observe the expression of Toll-like receptor 4 （TLR4） on the peripheral monocytes of ALI patients and c...Objective： To assess the effects of penehyclidine hydrochloride on patients with acute lung injury （ALI）, to observe the expression of Toll-like receptor 4 （TLR4） on the peripheral monocytes of ALI patients and changes of inflammatory ＆ anti-inflammatory cytokines and to investigate the mechanism of TLR4 in ALI.Methods： Forty-five patients with ALI were randomly divided into penehyclidine hydrochloride treatment group （P group, n=21） and conventional treatment group （control group, C group, n=24）. Patients in both groups received conventional treatment, including active treatment of the primary disease, respiratory support, nutritional support and fluid management therapy, while those in P group were given penehyclidine hydrochloride （1 mg, im, q. 12 h） in addition.The TLR4 expression of 20 healthy volunteers were detected.The clinical effect, average length of stay in ICU and hospital,values of PaO2 and PaO2/FiO2, expression of TLR4 on the surface of peripheral blood mononuclear cells and some serum cytokines were evaluated for 48 h.Results： The general conditions of the two groups were improved gradually and PaO2 increased progressively.Compared with 0 h, PaO2 and PaO2/FiO2 at 6, 12, 24 and 48 h after treatment were significantly increased （P＜0.05）. The improvement in P group was obviously greater than that in C group （P＜0.05）. The average length of hospitalization showed no difference between the two groups, but penehyclidine hydrochloride significantly decreased the average length of stay in ICU （t=3.485, P＜0.01）. The expression of TLR4 in two groups were both obviously higher than that of healthy volunteers （P＜0.01）. It decreased significantly at 24 h （t=2.032, P＜0.05） and 48 h （t=3.620, P＜0.01）and was lower in P group than in C group. The patients who showed a higher level of TLR4 expression in early stage had a worse prognosis and most of them developed acute respiratory distress syndrome （ARDS）. The incidence of ARDS was 23.8% in P group and 29.17% in C group at 24 h.Until148 h, there were other two patients developing ARDS in control group. Serum IL-l, IL-8 and TNF-α expressions reduced after 24 h in both groups. The reduction in P group was more obvious than that in C group （P＜0.05）. IL-13 increased gradually from 0 h to 24 h, and decreased slightly at 48 h, which showed no difference between two groups （t=1.028, P＞0.05）.Conclusions： Penehyclidine hydrochloride improves the arterial oxygen pressure, down-regulates the expression of TLR4 and restrains the inflammatory cytokines in the downstream of TLR4 signaling pathway. It prevents the development of ALI and can be considered as an important drug in ALI treatment.展开更多
基金the National Natural Science Foundation of China,No.39970719,30170919
文摘AIM: To determine the NF-kB activity in peripheral blood mononuclear cells (PBMC) in patients with acute cholangitis of severe type (ACST) and correlate the degree of NF-kB activation with severity of biliary tract infection and clinical outcome. METHODS: Twenty patients with ACST were divided into survivor group (13 cases) and nonsurvivor group (7 cases). Other ten patients undergoing elective gastrectomy or inguinal hernia repair were selected as control group. Peripheral blood samples were taken 24 hours postoperatively. PBMC were separated by density gradient centrifugation, then nuclear proteins were isolated from PBMC, and Electrophoretic Mobility Shift Assay (EMSA) used determined. The results were quantified by scanning densitometer of a Bio-Image Analysis System and expressed as relative optical density (ROD). The levels of TNF-alpha, IL-6, and IL-10 in the plasma of patients with ACST and healthy control subjects were determined by using an enzyme-linked immunoassay (ELISA). RESULTS: The NF-kB activity was 5.02 +/- 1.03 in nonsurvivor group, 2.98 +/- 0.51 in survivor group and 1.06 +/- 0.34 in control group. There were statistical differences in three groups (P【0.05). The levels of TNF-alpha and IL-6 in plasma were (498 +/- 53)ng.L(-1)and (587 +/- 64)ng.L(-1)in nonsurvivor group, (284 +/- 32)ng.L(-1) and (318 +/- 49)ng.L(-1)in survivor group and (89 +/- 11)ng.L(-1) and (102 +/-13)ng.L(-1)in control group. All patients with ACST had increased levels of TNF-alpha and IL-6, which were many-fold greater than those of control group, and there was an evidence of significantly higher levels in those of nonsurvivor group than that in survivor group (P【0.05). The levels of IL-10 in plasma were (378+/-32)ng.L(-1), (384+/-37)ng.L(-1) and (68+/-11)ng.L(-1) in three groups, respectively. All patients had also increased levels of IL-10 when compared with control group (P【0.05), but the IL-10 levels were not significantly higher in nonsurvivors than in survivors (P】0.05). CONCLUSION: NF-kB activity in PBMC in patients with ACST increases markedly and the degree of NF-kB activation is correlated with severity of biliary tract infection and clinical outcome.
基金Supported by the State Committee for Scientif ic Research (KBNgrant 4 PO5B 016 19
文摘AIM： To evaluate peripheral blood lymphocyte subsets in patients with acute pancreatitis （AP）. METHODS： Twenty patients with mild AP （M-AP） and 15 with severe AP （SAP） were included in our study. Peripheral blood lymphocytes were examined at d 1-3, 5, 10 and 30 by means of flow cytometry. RESULTS： A significant depletion of circulating lymphocytes was found in AP. In the early AP, the magnitude of depletion was similar for T- and B- lymphocytes. In the late course of S-AP, B-lymphocytes were much more depleted than T-lymphocytes. At d 10, strong shift in the CD7＋/CD19＋ ratio implicating predominance of T- over B-lymphocytes in S-AP was found. Among T-lymphocytes, the significant depletion of the CD4＋ population was observed in M-AP and S-AP, while CDS＋ cells were in the normal range. Lymphocytes were found to strongly express activation markers： CD69, CD25, CD28, CD38 and CD122. Serum interleukin-2 （IL-2）, IL-4, IL-5, IL-10, interferon-γ （IFN-γ） and tumor necrosis factor-α （TNF-α） levels were significantly increased in both forms of AP. The magnitude of elevation of cytokines known to be produced by Th2 was much higher than cytokines produced by Th1 cells. CONCLUSION： AP in humans is characterized by significant reduction of peripheral blood T- and B-lymphocytes.
基金Supported by the Grants From Shanghai Commission of Science and TechnologyShanghai Bureau of Health, No. 024Y32the grants from the Sino-German Center for Research Promotion, No.GZNr. 239(202/12)
文摘AIM： To investigate the role of inflammatory and anti-vira genes in the pathogenesis of SARS. METHODS： cDNA microarrays were used to screen the gene expression profiles of peripheral blood mononuclear cells （PBMCs） in two SARS patients （one in the acute severe phase and the other in the convalescent phase） and a healthy donor. In addition, real-time qualitative PCR was also performed to verify the reproducibility of the microarray results. The data were further analyzed. RESULTS： Many inflammatory and anti-viral genes were differentially expressed in SARS patients. Compared to the healthy control or the convalescent case, plenty of pro-inflammatory cytokines such as IL-1, TNF-α, IL-8, and MAPK signaling pathway were significantly upregulated in the acute severe case. However, anti-inflammatory agents such as IL-4 receptor, IL-13 receptor, IL-1Ra, and TNF-α-induced proteins 3 and 6 also increased dramatically in the acute severe case. On the contrary, a lot of IFN-stimulated genes like PKR, GBP-1 and 2, CXCL-10 and 11, and JAK/STAT signal pathway were downregulated in the acute severe case compared to the convalescent case. CONCLUSION： Gene expression in SAPS patients mirrors a host state of inflammation and anti-viral immunity at the transcription level, and understanding of gene expression profiles may make contribution to further studies of the SAPS pathogenesis.
基金Supported by Grants From Shanghai Committee of Scienceand Technology,Shanghai,China,No.044119624
文摘AIM:To study the effects of synthetic nonmethylated CpG-containing oligodeoxynucleotides(CpG-ODNs) ,either alone or combined with recombinant Hepatitis B surface antigen(HBsAg) polypeptide,on the phenotype,function,and intracellular signaling pathways of monocyte-derived dendritic cells(DCs) in patients with chronic hepatitis B(CHB) .METHODS:Peripheral blood monocytes isolated from CHB patients and healthy volunteers were induced to be dendritic cells by recombinant human granulocyte-monocyte colony stimulating factor and interleukin-4.The DCs were then treated with CpG-ODNs,CpGODNs/HBsAg,or tumor necrosis factor(TNF)-αfor 18 h.The expression of surface molecules including HLA-DR,CD86,and CD1a in DCs were detected by flow cytometry,and the expression of signal transducers and activators of transcription(STAT1,3,4,5,6) and suppressors of cell signaling(SOCS1,3) were determined by Western blotting assay.In addition,the capacity of DCs to stimulate allogeneic T lymphocytes and the amount of IL-12p70 released from DCs were measured.RESULTS:In the DCs derived from patients with CHB,treatment with TNF-α,CpG-ODNs,or CpG-ODNs/HBsAg,as compared to the vector control,significantly increased the expression of HLA-DR,stimulated the release of IL-12p70,and enhanced the capacity of DCs to stimulate allogenic T lymphocytes.The expressions of STAT1/4/6 and SOCS1/3,but not STAT3/5,were upregulated by TNF-α,CpG-ODNs,and CpG-ODNs/HBsAg.In addition,the expression of CD1a was upregulated only in the presence of both CpG-ODNs and HBsAg.CONCLUSION:The treatment with CpG-ODNs,either alone or combined with HBsAg,has a remarkable stimulatory effect on the impaired phenotype and function of DCs in CHB,possibly by regulating the expression of STAT1,4,6 and SOCS1,3.
文摘Objective： To assess the effects of penehyclidine hydrochloride on patients with acute lung injury （ALI）, to observe the expression of Toll-like receptor 4 （TLR4） on the peripheral monocytes of ALI patients and changes of inflammatory ＆ anti-inflammatory cytokines and to investigate the mechanism of TLR4 in ALI.Methods： Forty-five patients with ALI were randomly divided into penehyclidine hydrochloride treatment group （P group, n=21） and conventional treatment group （control group, C group, n=24）. Patients in both groups received conventional treatment, including active treatment of the primary disease, respiratory support, nutritional support and fluid management therapy, while those in P group were given penehyclidine hydrochloride （1 mg, im, q. 12 h） in addition.The TLR4 expression of 20 healthy volunteers were detected.The clinical effect, average length of stay in ICU and hospital,values of PaO2 and PaO2/FiO2, expression of TLR4 on the surface of peripheral blood mononuclear cells and some serum cytokines were evaluated for 48 h.Results： The general conditions of the two groups were improved gradually and PaO2 increased progressively.Compared with 0 h, PaO2 and PaO2/FiO2 at 6, 12, 24 and 48 h after treatment were significantly increased （P＜0.05）. The improvement in P group was obviously greater than that in C group （P＜0.05）. The average length of hospitalization showed no difference between the two groups, but penehyclidine hydrochloride significantly decreased the average length of stay in ICU （t=3.485, P＜0.01）. The expression of TLR4 in two groups were both obviously higher than that of healthy volunteers （P＜0.01）. It decreased significantly at 24 h （t=2.032, P＜0.05） and 48 h （t=3.620, P＜0.01）and was lower in P group than in C group. The patients who showed a higher level of TLR4 expression in early stage had a worse prognosis and most of them developed acute respiratory distress syndrome （ARDS）. The incidence of ARDS was 23.8% in P group and 29.17% in C group at 24 h.Until148 h, there were other two patients developing ARDS in control group. Serum IL-l, IL-8 and TNF-α expressions reduced after 24 h in both groups. The reduction in P group was more obvious than that in C group （P＜0.05）. IL-13 increased gradually from 0 h to 24 h, and decreased slightly at 48 h, which showed no difference between two groups （t=1.028, P＞0.05）.Conclusions： Penehyclidine hydrochloride improves the arterial oxygen pressure, down-regulates the expression of TLR4 and restrains the inflammatory cytokines in the downstream of TLR4 signaling pathway. It prevents the development of ALI and can be considered as an important drug in ALI treatment.
