By using an LKB-2277 Bioactivity Monitor, the effect ofporphyrin-cholesterol esters on cancer cells RL-60 were examined at 25℃.There is a dirrerence in the shape of the thermogenesls curve samong different kinds or ...By using an LKB-2277 Bioactivity Monitor, the effect ofporphyrin-cholesterol esters on cancer cells RL-60 were examined at 25℃.There is a dirrerence in the shape of the thermogenesls curve samong different kinds or Porphyrln-cholesterol esters,and the death rate or HL-60 cells is also different. The results show that porphyrin-chsterol esters can powerfully inhibit their metabolism, the inhibitory sequence is Ⅱ >I >Ⅲ,but the inhibitory way of each ester is different.展开更多
目的:建立检测胆固醇酯转运蛋白(cholesterol ester transfer protein,CETP)基因6种常见突变的等位基因特异性PCR技术。方法:应用针对CETP基因TaqIB(G→A)、I405V(A→G)、D442G(A→G)、R451Q(G→A)、A373P(G→C)和I14A(G→A)这6种常见...目的:建立检测胆固醇酯转运蛋白(cholesterol ester transfer protein,CETP)基因6种常见突变的等位基因特异性PCR技术。方法:应用针对CETP基因TaqIB(G→A)、I405V(A→G)、D442G(A→G)、R451Q(G→A)、A373P(G→C)和I14A(G→A)这6种常见的突变位点设计的等位基因特异性PCR技术,对海南汉、黎族人群中CETP基因突变类型进行了检测,同时对经上述等位基因特异性PCR检测的样本进行序列测定。结果:在海南汉、黎族人群中,TaqIB(G→A)突变位点可检测出GG、GA、AA3种基因型,I405V(A→G)突变位点可检测出AA、AG、GG3种基因型,D442G(A→G)突变位点可检测出AA、AG2种基因型,但在海南汉、黎族人群中未检测到R451Q(G→A)、A373P(G→C)和I14A(G→A)3种突变类型,用等位基因特异性PCR鉴定的CETP基因突变的基因分型结果与序列测定结果完全符合。结论:等位基因特异性PCR技术操作简便,重复性和稳定性好,可作为鉴定CETP基因突变类型的可行方法。展开更多
目的利用液相色谱-质谱联用法测定细胞内胆固醇及胆固醇酯,建立一种精确鉴定泡沫细胞的方法。方法用氧化修饰低密度脂蛋白(oxidized low density lipoprotein,oxLDL)干预U937巨噬细胞诱导泡沫细胞形成,然后用己烷/异丙醇(体积比3/2)抽...目的利用液相色谱-质谱联用法测定细胞内胆固醇及胆固醇酯,建立一种精确鉴定泡沫细胞的方法。方法用氧化修饰低密度脂蛋白(oxidized low density lipoprotein,oxLDL)干预U937巨噬细胞诱导泡沫细胞形成,然后用己烷/异丙醇(体积比3/2)抽提细胞内胆固醇及胆固醇酯,抽提液经真空干燥后再溶于甲醇,用液相色谱-质谱联用法直接测定细胞内游离胆固醇,胆固醇酯经胆固醇酯酶水解后测定总胆固醇,总胆固醇减去游离胆固醇即胆固醇酯。抽提脂质后的沉淀物溶于氢氧化钠溶液中,用BCA法测定蛋白含量,胆固醇及胆固醇酯用μg/mg细胞蛋白为单位来表示。结果氧化低密度脂蛋白干预后的巨噬细胞内存在高水平胆固醇及胆固醇酯的蓄积。在100μg的oxLDL处理组中,胆固醇酯与阴性对照组差异存在显著性[(25.733±2.970)μg/mg细胞蛋白vs (13.024±2.875)μg/mg细胞蛋白,P<0.01],同样胆固醇的水平差异也存在显著性[(18.675±1.315)μg/mg细胞蛋白vs (24.428±2.370)μg/mg细胞蛋白,P<0.01]。结论液相色谱-质谱联用法测定细胞内胆固醇是一种鉴定泡沫细胞简便而精确的方法。展开更多
文摘By using an LKB-2277 Bioactivity Monitor, the effect ofporphyrin-cholesterol esters on cancer cells RL-60 were examined at 25℃.There is a dirrerence in the shape of the thermogenesls curve samong different kinds or Porphyrln-cholesterol esters,and the death rate or HL-60 cells is also different. The results show that porphyrin-chsterol esters can powerfully inhibit their metabolism, the inhibitory sequence is Ⅱ >I >Ⅲ,but the inhibitory way of each ester is different.
基金We thank the National Natural Science Foundation of China （20872019） for the research financial support and we are grateful to Fudan University and Shanghai Institute of Organic Chemistry for recording EI-MS or ESI-MS, HRMS, ^1H NMR, and ^13C NMR spectra. We are also grateful to Dr. Hanqing Dong （OSI Pharma-ceuticals, USA） for his help in revising the manuscript.
文摘目的:建立检测胆固醇酯转运蛋白(cholesterol ester transfer protein,CETP)基因6种常见突变的等位基因特异性PCR技术。方法:应用针对CETP基因TaqIB(G→A)、I405V(A→G)、D442G(A→G)、R451Q(G→A)、A373P(G→C)和I14A(G→A)这6种常见的突变位点设计的等位基因特异性PCR技术,对海南汉、黎族人群中CETP基因突变类型进行了检测,同时对经上述等位基因特异性PCR检测的样本进行序列测定。结果:在海南汉、黎族人群中,TaqIB(G→A)突变位点可检测出GG、GA、AA3种基因型,I405V(A→G)突变位点可检测出AA、AG、GG3种基因型,D442G(A→G)突变位点可检测出AA、AG2种基因型,但在海南汉、黎族人群中未检测到R451Q(G→A)、A373P(G→C)和I14A(G→A)3种突变类型,用等位基因特异性PCR鉴定的CETP基因突变的基因分型结果与序列测定结果完全符合。结论:等位基因特异性PCR技术操作简便,重复性和稳定性好,可作为鉴定CETP基因突变类型的可行方法。
