AIM: To investigate production of TNFα from murine peritoneal macrophages stimulated with LPS, A 23187 , PMA, fMLP and fMLPP. METHODS: L929 cytotoxicity was used to show the level of released and cell associated TNF...AIM: To investigate production of TNFα from murine peritoneal macrophages stimulated with LPS, A 23187 , PMA, fMLP and fMLPP. METHODS: L929 cytotoxicity was used to show the level of released and cell associated TNFα from murine peritoneal macrophages measured by means of crystal violet staining assay. RESULTS: LPS(0 1～10 μg·mL -1 ) and A 23187 (0 01～1 μmol·L -1 ) were shown to induce a dose dependent increase of both released and cell associated TNFα, whereas, PMA(0 01～1 μmol·L -1 ), fMLP(0 025～2 5 μmol·L -1 ) and fMLP(0 025～2 5 μmol·L -1 ) did not induce macrophages to produce TNFα. CONCLUSION: LPS and A 23187 can induce production of TNFα from macrophages. Further research should be performed to study the pathways such as CD14 and calcium ion that may be involved in the biosynthesis of TNFα.展开更多
It has recently been established that neutrophils, the most abundant leukocytes, are capable of changes in gene expression during inflammatory responses. However, changes in the transcriptome as the neutrophil leaves ...It has recently been established that neutrophils, the most abundant leukocytes, are capable of changes in gene expression during inflammatory responses. However, changes in the transcriptome as the neutrophil leaves the bone marrow have yet to be described. We hypothesized that neutrophils are transcriptionally active cells that alter their gene expression profiles as they migrate into the vasculature and then into inflamed tissues. Our goal was to provide an overview of how the neutrophil＇s transcriptome changes as they migrate through different compartments using microarray and bio-informatic approaches. Our study demonstrates that neutrophils are highly plastic cells where normal environmental cues result in a site-specific neutrophil transcriptome. We demonstrate that neutrophil genes undergo one of four distinct expression change patterns as they move from bone marrow through the circulation to sites of inflammation： （i） continuously increasing; （ii） continuously decreasing; （iii） a down-up-down; and （iv） an up-down-up pattern. Additionally, we demonstrate that the neutrophil migration signaling network and the balance between anti-apoptotic and pro-apoptotic signaling are two of the main regulatory mechanisms that change as the neutrophil transits through compartments.展开更多
趋化抑制蛋白(Chemotaxis Inhibitory Protein ofStaphylococcus aureus,CHIPS)是由金黄色葡萄球菌分泌到菌体外的一种蛋白。在感染早期,它能特异性地与中性粒细胞和单核细胞上的C5a受体(C5aR)和fMLP受体(FPR)结合,从而阻止中性粒细胞...趋化抑制蛋白(Chemotaxis Inhibitory Protein ofStaphylococcus aureus,CHIPS)是由金黄色葡萄球菌分泌到菌体外的一种蛋白。在感染早期,它能特异性地与中性粒细胞和单核细胞上的C5a受体(C5aR)和fMLP受体(FPR)结合,从而阻止中性粒细胞和单核细胞对C5a和fMLP的结合作用,导致对病原吞噬作用的延迟。人们可以利用CHIPS对C5a-C5aR的阻止作用来研制治疗由C5a诱发的炎症性疾病的药物。将CHIPS作为免疫原防治疾病也将成为新的研究课题。实验成功构建了CHIPS蛋白的原核表达系统,为CHIPS免疫原性研究及蛋白的其他功能研究奠定了基础。展开更多
The presence of high reactive oxygen species (ROS) levels in semen is a major factor involved in the decline of male fertility. In seminal plasma, ROS are mainly produced by activated leucocytes. Spermatozoa were the ...The presence of high reactive oxygen species (ROS) levels in semen is a major factor involved in the decline of male fertility. In seminal plasma, ROS are mainly produced by activated leucocytes. Spermatozoa were the first cell type reported to show a potential susceptibility to oxidative damage. The aim of our study was to evaluate the impact of leucocytospermia on basal and FMLP (Formyl-Methionyl-Leucyl-Phenylalanine) induced oxidative status in semen of infertile men. We also analyzed the correlations of the spermatic parameters with amounts of ROS in semen. Our study included 50 semen samples of infertile men. Sperm analysis was performed using WHO standardized method. Seminal leucocytes were quantified using peroxidase technique. The measurement of ROS levels in semen was made by chemiluminescence assay. We measure respectively ROS amounts in neat semen and in washed sperm cells suspension from the same ejaculate. We also applied the test of provocation of leucocytes by FMLP on neat and washed samples to assess the spermatic oxidative status after leucocyte stimulation. Our results showed significant correlations between ROS levels in neat semen and many sperm parameters: motility, sperm concentration, leucocytes concentration and the rate of sperm cytoplasmic droplets. The studied samples were divided into 2 groups: (G1) composed of 36 samples without leucocytospermia and (G2) composed of 14 leucospermic samples. ROS levels were significantly lower in G1 than in G2 (p = 0.002). ROS production was significantly increased after application of FMLP in washed leucospermic samples (p = 0.001). The measurement of ROS in neat semen is a considerable contribution to explore the impairment of semen quality in infertile men. ROS levels in washed semen reflect the oxidative status generated by sperm preparation techniques used in assisted reproductive procedures. Levels of ROS are highly influenced by the presence of leucocytes and associated with decreased seminal parameters.展开更多
Objective: Chemotactic peptide may interfere with the process of tumor growth, invasion and metastasis by activating and attracting leukocytes containing macrophages. fMLP (CHO-Met-II e-Phe) is one of the chemotactic ...Objective: Chemotactic peptide may interfere with the process of tumor growth, invasion and metastasis by activating and attracting leukocytes containing macrophages. fMLP (CHO-Met-II e-Phe) is one of the chemotactic peptides. Boanmycin (BAM), a single A6 component from the bleomycin complex, is effective against a panel of cancers in clinical trials. This study was set to investigate the antitumor activity of BAM in combination with chemotactic peptide fMLP. Methods: Cytotoxicity of BAM and fMLP to cancer cells was determined by MTT assay. Therapeutic effect was evaluated by using the model of subcutaneously transplanted hepatoma 22 in mice. Results were judged as that a CDI less than 0.85 was considered as synergism and one less than 0.75 as significant synergism. Results: BAM and fMLP showed no synergism in cytotoxicity to cancer cells. In all in vivo experiments, fMLP was administered peritumorally at the dose of 1 mg/mouse; no significant inhibition by fMLP alone on the growth of hepatoma 22 was found. Different settings of BAM and fMLP combination included: (1) BAM, administered peritumorally×3, was started 24 h after tumor inoculation. BAM (0.5 mg/kg) alone and BAM-fMLP combination inhibited the growth of hepatoma 22 by 26.6% and 64.7%, respectively (P<0.05, CDI=0.36) on day 13. (2) BAM, administered ip×3, was started 24 h after tumor inoculation. The growth of tumor in BAM (1 mg/kg) group was faster than that in BAM-fMLP combination group. On day 14, BAM (1 mg/kg) alone and BAM-fMLP combination suppressed the growth of tumor by 11% and 70.6%, respectively (P<0.05), CDI=0.42). (3) BAM, administered ip×3, was started 96 h after tumor inoculation. The growth of tumor in BAM (1 mg/kg) group was faster than that in BAM-fMLP combination group. On day 13, BAM (1 mg/kg) alone and BAM-fMLP combination suppressed tumor growth by 38.2% and 77.1%, respectively (P<0.05, CDI=0.51). As shown in all in vivo experimental settings, antitumor effect of BAM in combination with fMLP was much more potent than that of BAM al展开更多
Objective： The inhibitory action of fMLP-boanmycin （BAM） combination on the growth of mouse colon carcinoma and its action mechanisms were observed in order to provide experimental proof for probing novel regimen o...Objective： The inhibitory action of fMLP-boanmycin （BAM） combination on the growth of mouse colon carcinoma and its action mechanisms were observed in order to provide experimental proof for probing novel regimen of chemotactic modulation in combination with chemotherapy in the treatment of cancer. Methods： Cytotoxicity of BAM-fMLP combination to tumor cells was determined by MTT assay in vitro. Antitumor activity of BAM-fMLP combination was assessed in mice subcutaneously transplanted colon carcinoma 26. The amount of superoxide anion （O2 ·^-）released from fMLP stimulated macrophages was determined by NBT assay. The amount of nitric oxide （NO） was indirectly determined by Griess method. Results： BAM-fMLP combination had no synergistic effect on tumor cells（CDI〉0.85）, but BAM at the doses of 10μg/ml, 30μg/ml and 100μg/ml in combination with fMLP at the concentration 20μg / ml exhibited synergistic effect on tumor cells in the presence of macrophages（CDI〈0.75）, fMLP inhibited the growth of colon carcinoma 26 by 50.0% when it at dose of 1 mg/mouse was administered peritumorally. BAM （1 mg/kg, intraperitoneally, three times） alone and BAM - fMLP combination inhibited the growth of colon carcinoma 26 by 38.6% and 78.4%, respectively （CDI=0.71） on day 12. The amount of O2 ·^- released from fMLP 4.6×10^-7 mol/L （0.2μg/ml） stimulated macrophages which were treated by BAM in vitro increased significantly（P〈0.01）, fMLP 2.3×10^-6 mol/L （1μg/ml） could not stimulate macrophages to release NO, but may stimulate macrophages treated with BAM 10μg/ml and 100μg/ml to release NO significantly（P〈0.01）. Conclusion： The inhibitory action of fMLP-boanmycin combination on the growth of mouse colon carcinoma have synergism, which may associate with the increase of O2 ·^- and NO released by macrophages. Chemotactic modulation in combination with chemotherapy may be a novel regimen in the treatment of cancer.展开更多
文摘AIM: To investigate production of TNFα from murine peritoneal macrophages stimulated with LPS, A 23187 , PMA, fMLP and fMLPP. METHODS: L929 cytotoxicity was used to show the level of released and cell associated TNFα from murine peritoneal macrophages measured by means of crystal violet staining assay. RESULTS: LPS(0 1～10 μg·mL -1 ) and A 23187 (0 01～1 μmol·L -1 ) were shown to induce a dose dependent increase of both released and cell associated TNFα, whereas, PMA(0 01～1 μmol·L -1 ), fMLP(0 025～2 5 μmol·L -1 ) and fMLP(0 025～2 5 μmol·L -1 ) did not induce macrophages to produce TNFα. CONCLUSION: LPS and A 23187 can induce production of TNFα from macrophages. Further research should be performed to study the pathways such as CD14 and calcium ion that may be involved in the biosynthesis of TNFα.
文摘It has recently been established that neutrophils, the most abundant leukocytes, are capable of changes in gene expression during inflammatory responses. However, changes in the transcriptome as the neutrophil leaves the bone marrow have yet to be described. We hypothesized that neutrophils are transcriptionally active cells that alter their gene expression profiles as they migrate into the vasculature and then into inflamed tissues. Our goal was to provide an overview of how the neutrophil＇s transcriptome changes as they migrate through different compartments using microarray and bio-informatic approaches. Our study demonstrates that neutrophils are highly plastic cells where normal environmental cues result in a site-specific neutrophil transcriptome. We demonstrate that neutrophil genes undergo one of four distinct expression change patterns as they move from bone marrow through the circulation to sites of inflammation： （i） continuously increasing; （ii） continuously decreasing; （iii） a down-up-down; and （iv） an up-down-up pattern. Additionally, we demonstrate that the neutrophil migration signaling network and the balance between anti-apoptotic and pro-apoptotic signaling are two of the main regulatory mechanisms that change as the neutrophil transits through compartments.
文摘趋化抑制蛋白(Chemotaxis Inhibitory Protein ofStaphylococcus aureus,CHIPS)是由金黄色葡萄球菌分泌到菌体外的一种蛋白。在感染早期,它能特异性地与中性粒细胞和单核细胞上的C5a受体(C5aR)和fMLP受体(FPR)结合,从而阻止中性粒细胞和单核细胞对C5a和fMLP的结合作用,导致对病原吞噬作用的延迟。人们可以利用CHIPS对C5a-C5aR的阻止作用来研制治疗由C5a诱发的炎症性疾病的药物。将CHIPS作为免疫原防治疾病也将成为新的研究课题。实验成功构建了CHIPS蛋白的原核表达系统,为CHIPS免疫原性研究及蛋白的其他功能研究奠定了基础。
文摘The presence of high reactive oxygen species (ROS) levels in semen is a major factor involved in the decline of male fertility. In seminal plasma, ROS are mainly produced by activated leucocytes. Spermatozoa were the first cell type reported to show a potential susceptibility to oxidative damage. The aim of our study was to evaluate the impact of leucocytospermia on basal and FMLP (Formyl-Methionyl-Leucyl-Phenylalanine) induced oxidative status in semen of infertile men. We also analyzed the correlations of the spermatic parameters with amounts of ROS in semen. Our study included 50 semen samples of infertile men. Sperm analysis was performed using WHO standardized method. Seminal leucocytes were quantified using peroxidase technique. The measurement of ROS levels in semen was made by chemiluminescence assay. We measure respectively ROS amounts in neat semen and in washed sperm cells suspension from the same ejaculate. We also applied the test of provocation of leucocytes by FMLP on neat and washed samples to assess the spermatic oxidative status after leucocyte stimulation. Our results showed significant correlations between ROS levels in neat semen and many sperm parameters: motility, sperm concentration, leucocytes concentration and the rate of sperm cytoplasmic droplets. The studied samples were divided into 2 groups: (G1) composed of 36 samples without leucocytospermia and (G2) composed of 14 leucospermic samples. ROS levels were significantly lower in G1 than in G2 (p = 0.002). ROS production was significantly increased after application of FMLP in washed leucospermic samples (p = 0.001). The measurement of ROS in neat semen is a considerable contribution to explore the impairment of semen quality in infertile men. ROS levels in washed semen reflect the oxidative status generated by sperm preparation techniques used in assisted reproductive procedures. Levels of ROS are highly influenced by the presence of leucocytes and associated with decreased seminal parameters.
基金This work was supported by the "973" Major State Basic Research Project of china (No. G1998051104)
文摘Objective: Chemotactic peptide may interfere with the process of tumor growth, invasion and metastasis by activating and attracting leukocytes containing macrophages. fMLP (CHO-Met-II e-Phe) is one of the chemotactic peptides. Boanmycin (BAM), a single A6 component from the bleomycin complex, is effective against a panel of cancers in clinical trials. This study was set to investigate the antitumor activity of BAM in combination with chemotactic peptide fMLP. Methods: Cytotoxicity of BAM and fMLP to cancer cells was determined by MTT assay. Therapeutic effect was evaluated by using the model of subcutaneously transplanted hepatoma 22 in mice. Results were judged as that a CDI less than 0.85 was considered as synergism and one less than 0.75 as significant synergism. Results: BAM and fMLP showed no synergism in cytotoxicity to cancer cells. In all in vivo experiments, fMLP was administered peritumorally at the dose of 1 mg/mouse; no significant inhibition by fMLP alone on the growth of hepatoma 22 was found. Different settings of BAM and fMLP combination included: (1) BAM, administered peritumorally×3, was started 24 h after tumor inoculation. BAM (0.5 mg/kg) alone and BAM-fMLP combination inhibited the growth of hepatoma 22 by 26.6% and 64.7%, respectively (P<0.05, CDI=0.36) on day 13. (2) BAM, administered ip×3, was started 24 h after tumor inoculation. The growth of tumor in BAM (1 mg/kg) group was faster than that in BAM-fMLP combination group. On day 14, BAM (1 mg/kg) alone and BAM-fMLP combination suppressed the growth of tumor by 11% and 70.6%, respectively (P<0.05), CDI=0.42). (3) BAM, administered ip×3, was started 96 h after tumor inoculation. The growth of tumor in BAM (1 mg/kg) group was faster than that in BAM-fMLP combination group. On day 13, BAM (1 mg/kg) alone and BAM-fMLP combination suppressed tumor growth by 38.2% and 77.1%, respectively (P<0.05, CDI=0.51). As shown in all in vivo experimental settings, antitumor effect of BAM in combination with fMLP was much more potent than that of BAM al
文摘Objective： The inhibitory action of fMLP-boanmycin （BAM） combination on the growth of mouse colon carcinoma and its action mechanisms were observed in order to provide experimental proof for probing novel regimen of chemotactic modulation in combination with chemotherapy in the treatment of cancer. Methods： Cytotoxicity of BAM-fMLP combination to tumor cells was determined by MTT assay in vitro. Antitumor activity of BAM-fMLP combination was assessed in mice subcutaneously transplanted colon carcinoma 26. The amount of superoxide anion （O2 ·^-）released from fMLP stimulated macrophages was determined by NBT assay. The amount of nitric oxide （NO） was indirectly determined by Griess method. Results： BAM-fMLP combination had no synergistic effect on tumor cells（CDI〉0.85）, but BAM at the doses of 10μg/ml, 30μg/ml and 100μg/ml in combination with fMLP at the concentration 20μg / ml exhibited synergistic effect on tumor cells in the presence of macrophages（CDI〈0.75）, fMLP inhibited the growth of colon carcinoma 26 by 50.0% when it at dose of 1 mg/mouse was administered peritumorally. BAM （1 mg/kg, intraperitoneally, three times） alone and BAM - fMLP combination inhibited the growth of colon carcinoma 26 by 38.6% and 78.4%, respectively （CDI=0.71） on day 12. The amount of O2 ·^- released from fMLP 4.6×10^-7 mol/L （0.2μg/ml） stimulated macrophages which were treated by BAM in vitro increased significantly（P〈0.01）, fMLP 2.3×10^-6 mol/L （1μg/ml） could not stimulate macrophages to release NO, but may stimulate macrophages treated with BAM 10μg/ml and 100μg/ml to release NO significantly（P〈0.01）. Conclusion： The inhibitory action of fMLP-boanmycin combination on the growth of mouse colon carcinoma have synergism, which may associate with the increase of O2 ·^- and NO released by macrophages. Chemotactic modulation in combination with chemotherapy may be a novel regimen in the treatment of cancer.
