The myeloid leukemic cell line HL-60 was studied by using DNA gel electrophoresis, flow cytomery, McAb C-myc, McAb Bc1-2 and CFU-L. From zero to 36 h,the apoptosis rates of 8 different phases and other indexes were ob...The myeloid leukemic cell line HL-60 was studied by using DNA gel electrophoresis, flow cytomery, McAb C-myc, McAb Bc1-2 and CFU-L. From zero to 36 h,the apoptosis rates of 8 different phases and other indexes were observed. The results showed that with the prolonged time of drug incubation,apoptosis of HL-60 cells increased progressively. This effect can be enhanced obviously by rh-IL-3 and rh-GM-CSF. At the same time,the killed rate of leukemic cells by Ara-C induction was increased. C-myc expression was decreased and Bc1-2 expression did not display apparent change. Interestingly, the normal hemopoietic cells were not affected by these two kinds of cytokine. The theoretical basis was provided for concurrent use of rh-IL-3, rh-GM-CSF and cytotoxic drugs whose purpose is to elevate remission rate during the phase of induced remission of leukemia.展开更多
Objective: To elucidate the pattern of chemo therapy drugs induced apoptosis and its role in chemo therapy of acute leukemia. Methods: Apoptosis induced by Ara C in human myeloid leukemia cell line HL 60 was inve...Objective: To elucidate the pattern of chemo therapy drugs induced apoptosis and its role in chemo therapy of acute leukemia. Methods: Apoptosis induced by Ara C in human myeloid leukemia cell line HL 60 was investigated by applying light microscope, electron microscopy combined with DNA electrophoresis and flow cytometry analysis techniques. Results: Apoptosis per sisted throughout 36 h following addition of Ara C with a gradual augmentation. Efficiency of apoptosis was enhanced in a dosedependent pattern, HL 60 treated with six other chemotherapy drugs and peripheral white blood cells from a AML case undergoing DA regimen chemo therapy exhibited typical DNA ladder pattern. Further investigation indicated that chemotherapy drugs apop tosis came into being possibly by downregulating the expression of c myc and bcl 2 oncogenes. Conclusion: Chemotherapy induced apoptosis is the primary mecha nism of chemotherapy.展开更多
为建立花生致敏蛋白Ara h 2双抗夹心酶联免疫吸附试验(ELISA)检测方法,本研究基于所制备的抗体,采用双抗体夹心检测模式,以Ara h 2鼠源单克隆抗体作为捕获抗体、兔源多克隆抗体作为检测抗体,并通过棋盘法优化抗体工作浓度,对该方法的灵...为建立花生致敏蛋白Ara h 2双抗夹心酶联免疫吸附试验(ELISA)检测方法,本研究基于所制备的抗体,采用双抗体夹心检测模式,以Ara h 2鼠源单克隆抗体作为捕获抗体、兔源多克隆抗体作为检测抗体,并通过棋盘法优化抗体工作浓度,对该方法的灵敏度、准确度、精密度和特异性进行鉴定。结果表明,建立的双抗夹心ELISA检测方法对Ara h 2的检出限为5.04 ng·mL^(-1),线性范围为15.63~1 000 ng·mL^(-1),添加回收率为80.12%~96.03%,批内和批间变异系数均小于10%,且特异性良好、与其他常见食物过敏原无交叉反应。本研究可为致敏蛋白Ara h 2检测提供一种快速高效的方法。展开更多
旨在为预防花生过敏提供新的思路和方法,考察了不同烘烤温度、烘烤时间和样品量(平铺于4 cm×4 cm锡箔纸)对Ara h 3抗原性的影响,利用正交试验得到Ara h 3抗原性增强程度最高的烘烤处理条件,并采用双缩脲法、紫外光谱、傅里叶变换...旨在为预防花生过敏提供新的思路和方法,考察了不同烘烤温度、烘烤时间和样品量(平铺于4 cm×4 cm锡箔纸)对Ara h 3抗原性的影响,利用正交试验得到Ara h 3抗原性增强程度最高的烘烤处理条件,并采用双缩脲法、紫外光谱、傅里叶变换红外光谱和荧光光谱探究烘烤对Ara h 3溶解度、游离巯基含量、二级结构和疏水性的影响。结果表明:烘烤后Ara h 3抗原性增强,且随烘烤温度升高和烘烤时间延长Ara h 3的抗原性呈先增强后减弱的趋势,样品量在70~90 mg时抗原性相差不大;在烘烤温度140℃、烘烤时间20 min,样品量80 mg时,抗原性增强程度最高,达到34.12%;烘烤后Ara h 3溶解度和游离巯基含量显著降低,蛋白质发生解折叠,无规卷曲含量增加,蛋白质分子趋于无序状态,同时,蛋白质部分肽链舒展,疏水基团暴露,外源荧光强度增强。综上,烘烤处理可以改变Ara h 3的结构,使其抗原表位暴露或者产生新的抗原表位,显著增强Ara h 3的抗原性。花生加工过程中应规避致使花生抗原性增强的烘烤条件,以降低花生致敏性。展开更多
花生过敏蛋白Ara h 3广泛存在于各类花生制品中,少量摄入即可引发过敏反应,严重威胁花生过敏患者的健康。精准地识别Ara h 3便于开展后续免疫学检测,因此,制备高纯度、高特异性的抗体显得尤为重要。以花生脱脂粉为原料,通过HiTrap Q HP...花生过敏蛋白Ara h 3广泛存在于各类花生制品中,少量摄入即可引发过敏反应,严重威胁花生过敏患者的健康。精准地识别Ara h 3便于开展后续免疫学检测,因此,制备高纯度、高特异性的抗体显得尤为重要。以花生脱脂粉为原料,通过HiTrap Q HP阴离子交换层析柱和HiLoad Superdex 200 pg制备级分子筛层析柱,制备高纯度的花生致敏蛋白Ara h 3,将其与CNBr-activated SepharoseTM 4B进行偶联,利用不同洗脱液对血清样品中的非特异性及特异性物质进行分离,制备高纯度、高特异性的抗Ara h 3多克隆抗体。结果表明:Gly-HCl洗脱的蛋白质量浓度为0.6 mg/mL;SDS-PAGE分析显示,多克隆抗体中的非特异性物质已被有效去除;免疫印迹和ELISA试验证明,纯化后的抗体仍具有较高的特异性,其效价达到1∶51200。上述结果表明抗体成功纯化,为花生过敏原Ara h 3的检测研究奠定了基础。展开更多
文摘The myeloid leukemic cell line HL-60 was studied by using DNA gel electrophoresis, flow cytomery, McAb C-myc, McAb Bc1-2 and CFU-L. From zero to 36 h,the apoptosis rates of 8 different phases and other indexes were observed. The results showed that with the prolonged time of drug incubation,apoptosis of HL-60 cells increased progressively. This effect can be enhanced obviously by rh-IL-3 and rh-GM-CSF. At the same time,the killed rate of leukemic cells by Ara-C induction was increased. C-myc expression was decreased and Bc1-2 expression did not display apparent change. Interestingly, the normal hemopoietic cells were not affected by these two kinds of cytokine. The theoretical basis was provided for concurrent use of rh-IL-3, rh-GM-CSF and cytotoxic drugs whose purpose is to elevate remission rate during the phase of induced remission of leukemia.
文摘Objective: To elucidate the pattern of chemo therapy drugs induced apoptosis and its role in chemo therapy of acute leukemia. Methods: Apoptosis induced by Ara C in human myeloid leukemia cell line HL 60 was investigated by applying light microscope, electron microscopy combined with DNA electrophoresis and flow cytometry analysis techniques. Results: Apoptosis per sisted throughout 36 h following addition of Ara C with a gradual augmentation. Efficiency of apoptosis was enhanced in a dosedependent pattern, HL 60 treated with six other chemotherapy drugs and peripheral white blood cells from a AML case undergoing DA regimen chemo therapy exhibited typical DNA ladder pattern. Further investigation indicated that chemotherapy drugs apop tosis came into being possibly by downregulating the expression of c myc and bcl 2 oncogenes. Conclusion: Chemotherapy induced apoptosis is the primary mecha nism of chemotherapy.
文摘为建立花生致敏蛋白Ara h 2双抗夹心酶联免疫吸附试验(ELISA)检测方法,本研究基于所制备的抗体,采用双抗体夹心检测模式,以Ara h 2鼠源单克隆抗体作为捕获抗体、兔源多克隆抗体作为检测抗体,并通过棋盘法优化抗体工作浓度,对该方法的灵敏度、准确度、精密度和特异性进行鉴定。结果表明,建立的双抗夹心ELISA检测方法对Ara h 2的检出限为5.04 ng·mL^(-1),线性范围为15.63~1 000 ng·mL^(-1),添加回收率为80.12%~96.03%,批内和批间变异系数均小于10%,且特异性良好、与其他常见食物过敏原无交叉反应。本研究可为致敏蛋白Ara h 2检测提供一种快速高效的方法。
文摘旨在为预防花生过敏提供新的思路和方法,考察了不同烘烤温度、烘烤时间和样品量(平铺于4 cm×4 cm锡箔纸)对Ara h 3抗原性的影响,利用正交试验得到Ara h 3抗原性增强程度最高的烘烤处理条件,并采用双缩脲法、紫外光谱、傅里叶变换红外光谱和荧光光谱探究烘烤对Ara h 3溶解度、游离巯基含量、二级结构和疏水性的影响。结果表明:烘烤后Ara h 3抗原性增强,且随烘烤温度升高和烘烤时间延长Ara h 3的抗原性呈先增强后减弱的趋势,样品量在70~90 mg时抗原性相差不大;在烘烤温度140℃、烘烤时间20 min,样品量80 mg时,抗原性增强程度最高,达到34.12%;烘烤后Ara h 3溶解度和游离巯基含量显著降低,蛋白质发生解折叠,无规卷曲含量增加,蛋白质分子趋于无序状态,同时,蛋白质部分肽链舒展,疏水基团暴露,外源荧光强度增强。综上,烘烤处理可以改变Ara h 3的结构,使其抗原表位暴露或者产生新的抗原表位,显著增强Ara h 3的抗原性。花生加工过程中应规避致使花生抗原性增强的烘烤条件,以降低花生致敏性。
文摘花生过敏蛋白Ara h 3广泛存在于各类花生制品中,少量摄入即可引发过敏反应,严重威胁花生过敏患者的健康。精准地识别Ara h 3便于开展后续免疫学检测,因此,制备高纯度、高特异性的抗体显得尤为重要。以花生脱脂粉为原料,通过HiTrap Q HP阴离子交换层析柱和HiLoad Superdex 200 pg制备级分子筛层析柱,制备高纯度的花生致敏蛋白Ara h 3,将其与CNBr-activated SepharoseTM 4B进行偶联,利用不同洗脱液对血清样品中的非特异性及特异性物质进行分离,制备高纯度、高特异性的抗Ara h 3多克隆抗体。结果表明:Gly-HCl洗脱的蛋白质量浓度为0.6 mg/mL;SDS-PAGE分析显示,多克隆抗体中的非特异性物质已被有效去除;免疫印迹和ELISA试验证明,纯化后的抗体仍具有较高的特异性,其效价达到1∶51200。上述结果表明抗体成功纯化,为花生过敏原Ara h 3的检测研究奠定了基础。
