目的研究中药金银花水提物(以下简称金银花)对卵清蛋白(OVA)致敏小鼠的免疫调控作用,探索中药治疗食物过敏的可行性。方法取BALB/c小鼠40只,均分为5组,每组8只,按照肠道激发后灌服不同浓度的金银花水提物分为100 mg/100 m l(H)、50 mg/1...目的研究中药金银花水提物(以下简称金银花)对卵清蛋白(OVA)致敏小鼠的免疫调控作用,探索中药治疗食物过敏的可行性。方法取BALB/c小鼠40只,均分为5组,每组8只,按照肠道激发后灌服不同浓度的金银花水提物分为100 mg/100 m l(H)、50 mg/100 m l(M)、25 mg/100 m l(L)浓度组,激发后不给药组(Ch),以及正常生理盐水(NS)对照组。取空肠行HE及甲苯胺蓝染色;组织荧光法测定小肠组胺含量;ELISA法测定外周淋巴组织单个核细胞(PLNMC)培养上清液中白细胞介素4(IL-4)、γ干扰素(IFN-γ)及血清OVA特异性IgE(OVA-sIgE)水平;RT-PCR测定PLNMC中IL-12p40 mRNA表达;足垫肿胀实验检测迟发型超敏反应。结果H、M组浓度的金银花水提物可缓解过敏小鼠小肠绒毛炎症,减轻肥大细胞聚集和脱颗粒,提高固有层完整肥大细胞比率,减轻过敏小鼠肠道组胺释放,降低过敏小鼠体内IL-4、OVA-sIgE水平及IL-4/IFN-γ比值,抑制PLNMC中IL-12 mRNA表达;三种浓度金银花可缓解OVA介导的小鼠足垫迟发性超敏反应(DTH)。结论证实金银花水提物可参与OVA致敏小鼠的免疫调控,对缓解OVA介导的小鼠IgE及细胞介导的变态反应有利,用于食物过敏治疗具有潜在研究价值。展开更多
OBJECTIVE: To investigate genes involved in cancer metastasis, mRNA differential display was used to compare the levels of gene expression in two cell sublines derived from human giant cell carcinoma of lung (PG) whic...OBJECTIVE: To investigate genes involved in cancer metastasis, mRNA differential display was used to compare the levels of gene expression in two cell sublines derived from human giant cell carcinoma of lung (PG) which had different metastatic potentials. METHODS: Using in vivo tumorigenicity and a spontaneous metastasis assay in nude mice, two sublines (BE1, LH7) from human giant cell carcinoma of lung (PG) with different metastatic potentials were isolated and characterized. mRNA differential display was used to compare the levels of gene expression between them and the obtained results were confirmed by Northern hybridization. RESULTS: One differentially expressed band was nearly identical (99% homology) to Ras-GTPase-Activating protein SH3 domain binding protein (G3BP). G3BP displayed a strong expression in LH7 (non-metastatic in recipient nude mice) and a very weak expression in BE1 (100% metastatic frequency). The same different expression level of G3BP was detected in Northern hybridization with another panel of cell sublines with different metastatic potentials (established in our lab) derived from human prostate carcinoma cell line PC-3M. CONCLUSION: Our results indicate that G3BP was implicated in cancer metastasis because of its differential expressions in the two panels of cell sublines with different metastatic potentials.展开更多
OBJECTIVE: To investigate an method for hepatic differentiation from embryonic stem cells (ES cells) in vitro and the resulting differentiation ratio, in order to develop a procedure for producing a new type of hepato...OBJECTIVE: To investigate an method for hepatic differentiation from embryonic stem cells (ES cells) in vitro and the resulting differentiation ratio, in order to develop a procedure for producing a new type of hepatocyte for hepatocyte replacement therapy in the treatment of liver failure. METHODS: ES cells from Balb/C mice were cultured and maintained in an undifferentiated state in gelatin-coated dishes using Dulbecco's modified Eagle's medium (DMEM) containing 1000 U/ml leukemia inhibitory factor (LIF). Then, LIF was withdrawn from the DMEM to allow the ES cells to develop into embryonic bodies (EBs). EBs were plated onto tissue culture dishes, and growth factors such as acidicfibroblast growth factor (aFGF) and hepatocyte growth factor (HGF) were added to the medium to promote directional differentiation. The course of development and differentiation was observed dynamically using an inversion microscope. The expression of hepatic proteins, such as alpha-fetoprotein (AFP), albumin (ALB), cytokeratin 8 (CK8), cytokeratin 18 (CK18), in cytoplasm was analyzed by immunocytochemistry (ICC). The concentration of ALB in the medium was determined dynamically by radioimmunoassay (RIA). RESULTS: ES cells replicated as clones, without differentiating, in DMEM containing LIF. They developed into EBs in medium without LIF. Our ICC assay showed that differentiating cells did not express hepatic proteins, such as AFP, ALB, CK8, and CK18 until day 7, day 9, day 11, and day 11, respectively (up to 2 days later when growth factors are not present). The concentration of AFP in the medium was first detected on day 8, at a concentration of 3.4 ng/ml, and increased to 22.8 ng/ml by day 15. The concentration of ALB in the medium was 0.2 micro g/ml on day 11, and increased to 2.2 micro g/ml by day 15. ALB-positive cells under ICC manifest morphological structures were consistent with normal mouse hepatocytes. The differentiation ratio of hepatocytes in the ES cell differentiation system was 30% on day 15 (significantly lower withou展开更多
文摘目的研究中药金银花水提物(以下简称金银花)对卵清蛋白(OVA)致敏小鼠的免疫调控作用,探索中药治疗食物过敏的可行性。方法取BALB/c小鼠40只,均分为5组,每组8只,按照肠道激发后灌服不同浓度的金银花水提物分为100 mg/100 m l(H)、50 mg/100 m l(M)、25 mg/100 m l(L)浓度组,激发后不给药组(Ch),以及正常生理盐水(NS)对照组。取空肠行HE及甲苯胺蓝染色;组织荧光法测定小肠组胺含量;ELISA法测定外周淋巴组织单个核细胞(PLNMC)培养上清液中白细胞介素4(IL-4)、γ干扰素(IFN-γ)及血清OVA特异性IgE(OVA-sIgE)水平;RT-PCR测定PLNMC中IL-12p40 mRNA表达;足垫肿胀实验检测迟发型超敏反应。结果H、M组浓度的金银花水提物可缓解过敏小鼠小肠绒毛炎症,减轻肥大细胞聚集和脱颗粒,提高固有层完整肥大细胞比率,减轻过敏小鼠肠道组胺释放,降低过敏小鼠体内IL-4、OVA-sIgE水平及IL-4/IFN-γ比值,抑制PLNMC中IL-12 mRNA表达;三种浓度金银花可缓解OVA介导的小鼠足垫迟发性超敏反应(DTH)。结论证实金银花水提物可参与OVA致敏小鼠的免疫调控,对缓解OVA介导的小鼠IgE及细胞介导的变态反应有利,用于食物过敏治疗具有潜在研究价值。
基金86 3HighTechnologyProject (No # 10 2 10 0 1 0 9)andSpecialFoundationforPh DTrainingProgramofEducationMinistry (No 19990 0
文摘OBJECTIVE: To investigate genes involved in cancer metastasis, mRNA differential display was used to compare the levels of gene expression in two cell sublines derived from human giant cell carcinoma of lung (PG) which had different metastatic potentials. METHODS: Using in vivo tumorigenicity and a spontaneous metastasis assay in nude mice, two sublines (BE1, LH7) from human giant cell carcinoma of lung (PG) with different metastatic potentials were isolated and characterized. mRNA differential display was used to compare the levels of gene expression between them and the obtained results were confirmed by Northern hybridization. RESULTS: One differentially expressed band was nearly identical (99% homology) to Ras-GTPase-Activating protein SH3 domain binding protein (G3BP). G3BP displayed a strong expression in LH7 (non-metastatic in recipient nude mice) and a very weak expression in BE1 (100% metastatic frequency). The same different expression level of G3BP was detected in Northern hybridization with another panel of cell sublines with different metastatic potentials (established in our lab) derived from human prostate carcinoma cell line PC-3M. CONCLUSION: Our results indicate that G3BP was implicated in cancer metastasis because of its differential expressions in the two panels of cell sublines with different metastatic potentials.
基金ThisworkwassupportedbytheNationalKeyResearchFoundationofChina ( 973 ) (No 2 0 0 1CB5 10 10 1)andtheNationalNaturalScienceFoundationofChina (No 3 0 2 3 0 3 5 0 No 60 2 780 14 )
文摘OBJECTIVE: To investigate an method for hepatic differentiation from embryonic stem cells (ES cells) in vitro and the resulting differentiation ratio, in order to develop a procedure for producing a new type of hepatocyte for hepatocyte replacement therapy in the treatment of liver failure. METHODS: ES cells from Balb/C mice were cultured and maintained in an undifferentiated state in gelatin-coated dishes using Dulbecco's modified Eagle's medium (DMEM) containing 1000 U/ml leukemia inhibitory factor (LIF). Then, LIF was withdrawn from the DMEM to allow the ES cells to develop into embryonic bodies (EBs). EBs were plated onto tissue culture dishes, and growth factors such as acidicfibroblast growth factor (aFGF) and hepatocyte growth factor (HGF) were added to the medium to promote directional differentiation. The course of development and differentiation was observed dynamically using an inversion microscope. The expression of hepatic proteins, such as alpha-fetoprotein (AFP), albumin (ALB), cytokeratin 8 (CK8), cytokeratin 18 (CK18), in cytoplasm was analyzed by immunocytochemistry (ICC). The concentration of ALB in the medium was determined dynamically by radioimmunoassay (RIA). RESULTS: ES cells replicated as clones, without differentiating, in DMEM containing LIF. They developed into EBs in medium without LIF. Our ICC assay showed that differentiating cells did not express hepatic proteins, such as AFP, ALB, CK8, and CK18 until day 7, day 9, day 11, and day 11, respectively (up to 2 days later when growth factors are not present). The concentration of AFP in the medium was first detected on day 8, at a concentration of 3.4 ng/ml, and increased to 22.8 ng/ml by day 15. The concentration of ALB in the medium was 0.2 micro g/ml on day 11, and increased to 2.2 micro g/ml by day 15. ALB-positive cells under ICC manifest morphological structures were consistent with normal mouse hepatocytes. The differentiation ratio of hepatocytes in the ES cell differentiation system was 30% on day 15 (significantly lower withou
