目的:研究染色体8q24区段的rs987525位点及MAFB基因的rs13041247位点单核苷酸多态性(single nucleotide polymorphism,SNP)与非综合征型唇腭裂(nonsyndromic cleft lip with or without cleft palate,NSCL/P)的相关性。方法:收集宁夏地...目的:研究染色体8q24区段的rs987525位点及MAFB基因的rs13041247位点单核苷酸多态性(single nucleotide polymorphism,SNP)与非综合征型唇腭裂(nonsyndromic cleft lip with or without cleft palate,NSCL/P)的相关性。方法:收集宁夏地区病例组非综合征型唇腭裂患儿369例,患儿父亲303例,母亲311例,完整3人核心家系158个。对照组收集在校正常大学生433例。采用聚合酶链反应-限制性片段长度多态性(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)的方法检测这2个SNPs的基因型,运用卡方检验,传递不平衡检验(transmission disequilibrium test,TDT)等统计学方法分析评价以上SNPs的基因型频率和等位基因频率在患儿,患儿父母及对照组人群中的分布,评价其与NSCL/P的关联。结果:病例对照研究发现:MAFB基因的rs13041247位点在本研究人群的单纯唇裂组及唇裂伴或不伴腭裂组中基因型与等位基因频率与对照组比较存在统计学差异(P=0.02,P=0.04,P=0.01,P=0.04),而在单纯腭裂组中不存在统计学差异(P=0.25,P=0.51)。染色体8q24区域的rs987525位点基因型与等位基因频率与对照组比较差异无统计学意义(P=0.07,P=0.20,P=0.58,P=0.33,P=0.35,P=0.59)。TDT发现rs13041247位点C等位基因在单纯唇裂组中存在过传递(P=0.03),rs987525位点A等位基因在唇裂,腭裂及唇裂伴或不伴腭裂组中存在过传递(P=0.00,P=0.04,P=0.00)。结论:MAFB的rs13041247位点单核苷酸多态性与非综合征型唇腭裂有关。展开更多
[目的]研究非小细胞肺癌(NSCLC)患者中转录因子MafB的表达变化,并评估MafB对NSCLC患者CD14^+单核细胞分泌Ⅰ型干扰素和诱导CD4^+T细胞分化的影响。[方法]入组41例NSCLC患者(28例鳞癌和13例腺癌)和21例健康志愿者。收集外周血和支气管肺...[目的]研究非小细胞肺癌(NSCLC)患者中转录因子MafB的表达变化,并评估MafB对NSCLC患者CD14^+单核细胞分泌Ⅰ型干扰素和诱导CD4^+T细胞分化的影响。[方法]入组41例NSCLC患者(28例鳞癌和13例腺癌)和21例健康志愿者。收集外周血和支气管肺泡灌洗液(BALF),分离血浆和外周血单个核细胞(PBMC),纯化CD14^+单核细胞和CD4^+T细胞。酶联免疫吸附实验检测血浆和BALF干扰素(IFN)-α和IFN-β的水平,反转录实时定量PCR检测外周血和BALF中MafBm RNA相对表达量,Western blot检测MafB蛋白水平。利用MafBsi RNA转染CD14^+单核细胞,观察抑制MafB对CD14^+单核细胞分泌IFN-α和IFN-β的影响,检测干扰素调节因子3(IRF3)磷酸化水平。建立CD14^+单核细胞和CD4^+T细胞的直接接触和间接接触培养系统,通过ELISA法检测培养上清中IFN-γ、白细胞介素(IL)-4、IL-17和IL-21表达水平评估抑制MafB对CD14^+单核细胞调控CD4^+T细胞分化的影响。[结果]血浆IFN-α和IFN-β水平在健康志愿者和NSCLC患者之间差异无统计学意义,但肿瘤部位BALF中IFN-α(242.5±59.98pg/ml vs 282.5±45.24pg/ml,P=0.013)和IFN-β(12.40±2.81pg/ml vs 34.42±7.83pg/ml,P<0.0001)水平均显著性低于非肿瘤部位。PBMC中MafBm RNA相对表达量和蛋白水平在健康志愿者和NSCLC患者之间差异亦无统计学意义,但肿瘤部位BALF中MafB m RNA和蛋白水平则显著性高于非肿瘤部位(P<0.0001)。MafBsi RNA转染可显著性抑制CD14^+单核细胞中MafBm RNA和蛋白的表达。MafBsi RNA转染可促进CD14^+单核细胞IFN-β的分泌(16.09±5.79pg/ml vs 6.73±1.78pg/ml,P<0.0001),增加IRF3磷酸化(P<0.0001),但对IFN-α表达无明显影响(P>0.05)。而抑制NSCLC患者CD14^+单核细胞中MafB对CD14^+单核细胞和CD4^+T细胞共培养系统中IFN-γ、IL-4、IL-17和IL-21的分泌水平并无显著性影响(P>0.05)。[结论]NSCLC患者肿瘤部位过度表达的MafB可能诱导了Ⅰ型干扰素抑制,但MafB对肿瘤部位CD14^+单核细胞的免疫调控功能可能无影响。展开更多
Background In previous work, we suggested that some 2aminosteroids inhibited proliferation and induced differentiation of both human and murine leukemia cells Here, we reported the actions of another new 2aminosteroid...Background In previous work, we suggested that some 2aminosteroids inhibited proliferation and induced differentiation of both human and murine leukemia cells Here, we reported the actions of another new 2aminosteroid designated as H89712 on human leukemia cells Methods Cell colony counting and MTT assay were used to determine proliferation Cell morphology, histochemical staining, UV detection and cytometry were used to determine differentiation RTPCR was used to detect gene expression. Standard statistical method was used to analyze dataResults H89712 inhibited proliferation of HL60 leukemia cells and the inhibition percentage in MTT assay was 18% at the dose of 108 mol/L and 65% at the dose of 105 mol/L, respectively The inhibition for HL60 in colony assay was 23% at the dose of 108 mol/L and 96% at the dose of 105 mol/L, respectively H89712 also induced HL60 cells toward macrophagelike differentiation It was verified by flow cytometry that the percentage of positive CD14 expression in differentiated HL60 cells was about 9 times higher than that of the control at the dose of 108 mol/L and 20 times higher than that of the control at the dose of 105 mol/L respectively, and this action involved upregulation of MafB gene in HL60 leukemia cells On the other hand, H89712 inhibited proliferation of K562 leukemia cells and the inhibition of K562 leukemia cells in MTT assay was shown by 34% at the dose of 108 mol/L and 88% at the dose of 105 mol/L respectively The inhibition of K562 leukemia cells in colony assay was 53% at the dose of 108 mol/L and 100% at the dose of 105 mol/L respectively H89712 also induced K562 cells toward erythroidlike differentiation and it was verified by flow cytometry that the percentage of positive CD71 expression in differentiated K562 cells was about 9 times higher than that of the control at the dose of 108 mol/L and 16 times higher than that of the control at the dose of 105 mol/L respectively This action was related to upregulation of Egr1 gene in K562 leukemia cellsConclusions Our results showed the important roles played by MafB in macrophage differentiation and Egr1 in erythroid differentiation of human myeloid leukemia cells展开更多
文摘目的:研究染色体8q24区段的rs987525位点及MAFB基因的rs13041247位点单核苷酸多态性(single nucleotide polymorphism,SNP)与非综合征型唇腭裂(nonsyndromic cleft lip with or without cleft palate,NSCL/P)的相关性。方法:收集宁夏地区病例组非综合征型唇腭裂患儿369例,患儿父亲303例,母亲311例,完整3人核心家系158个。对照组收集在校正常大学生433例。采用聚合酶链反应-限制性片段长度多态性(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)的方法检测这2个SNPs的基因型,运用卡方检验,传递不平衡检验(transmission disequilibrium test,TDT)等统计学方法分析评价以上SNPs的基因型频率和等位基因频率在患儿,患儿父母及对照组人群中的分布,评价其与NSCL/P的关联。结果:病例对照研究发现:MAFB基因的rs13041247位点在本研究人群的单纯唇裂组及唇裂伴或不伴腭裂组中基因型与等位基因频率与对照组比较存在统计学差异(P=0.02,P=0.04,P=0.01,P=0.04),而在单纯腭裂组中不存在统计学差异(P=0.25,P=0.51)。染色体8q24区域的rs987525位点基因型与等位基因频率与对照组比较差异无统计学意义(P=0.07,P=0.20,P=0.58,P=0.33,P=0.35,P=0.59)。TDT发现rs13041247位点C等位基因在单纯唇裂组中存在过传递(P=0.03),rs987525位点A等位基因在唇裂,腭裂及唇裂伴或不伴腭裂组中存在过传递(P=0.00,P=0.04,P=0.00)。结论:MAFB的rs13041247位点单核苷酸多态性与非综合征型唇腭裂有关。
文摘[目的]研究非小细胞肺癌(NSCLC)患者中转录因子MafB的表达变化,并评估MafB对NSCLC患者CD14^+单核细胞分泌Ⅰ型干扰素和诱导CD4^+T细胞分化的影响。[方法]入组41例NSCLC患者(28例鳞癌和13例腺癌)和21例健康志愿者。收集外周血和支气管肺泡灌洗液(BALF),分离血浆和外周血单个核细胞(PBMC),纯化CD14^+单核细胞和CD4^+T细胞。酶联免疫吸附实验检测血浆和BALF干扰素(IFN)-α和IFN-β的水平,反转录实时定量PCR检测外周血和BALF中MafBm RNA相对表达量,Western blot检测MafB蛋白水平。利用MafBsi RNA转染CD14^+单核细胞,观察抑制MafB对CD14^+单核细胞分泌IFN-α和IFN-β的影响,检测干扰素调节因子3(IRF3)磷酸化水平。建立CD14^+单核细胞和CD4^+T细胞的直接接触和间接接触培养系统,通过ELISA法检测培养上清中IFN-γ、白细胞介素(IL)-4、IL-17和IL-21表达水平评估抑制MafB对CD14^+单核细胞调控CD4^+T细胞分化的影响。[结果]血浆IFN-α和IFN-β水平在健康志愿者和NSCLC患者之间差异无统计学意义,但肿瘤部位BALF中IFN-α(242.5±59.98pg/ml vs 282.5±45.24pg/ml,P=0.013)和IFN-β(12.40±2.81pg/ml vs 34.42±7.83pg/ml,P<0.0001)水平均显著性低于非肿瘤部位。PBMC中MafBm RNA相对表达量和蛋白水平在健康志愿者和NSCLC患者之间差异亦无统计学意义,但肿瘤部位BALF中MafB m RNA和蛋白水平则显著性高于非肿瘤部位(P<0.0001)。MafBsi RNA转染可显著性抑制CD14^+单核细胞中MafBm RNA和蛋白的表达。MafBsi RNA转染可促进CD14^+单核细胞IFN-β的分泌(16.09±5.79pg/ml vs 6.73±1.78pg/ml,P<0.0001),增加IRF3磷酸化(P<0.0001),但对IFN-α表达无明显影响(P>0.05)。而抑制NSCLC患者CD14^+单核细胞中MafB对CD14^+单核细胞和CD4^+T细胞共培养系统中IFN-γ、IL-4、IL-17和IL-21的分泌水平并无显著性影响(P>0.05)。[结论]NSCLC患者肿瘤部位过度表达的MafB可能诱导了Ⅰ型干扰素抑制,但MafB对肿瘤部位CD14^+单核细胞的免疫调控功能可能无影响。
文摘Background In previous work, we suggested that some 2aminosteroids inhibited proliferation and induced differentiation of both human and murine leukemia cells Here, we reported the actions of another new 2aminosteroid designated as H89712 on human leukemia cells Methods Cell colony counting and MTT assay were used to determine proliferation Cell morphology, histochemical staining, UV detection and cytometry were used to determine differentiation RTPCR was used to detect gene expression. Standard statistical method was used to analyze dataResults H89712 inhibited proliferation of HL60 leukemia cells and the inhibition percentage in MTT assay was 18% at the dose of 108 mol/L and 65% at the dose of 105 mol/L, respectively The inhibition for HL60 in colony assay was 23% at the dose of 108 mol/L and 96% at the dose of 105 mol/L, respectively H89712 also induced HL60 cells toward macrophagelike differentiation It was verified by flow cytometry that the percentage of positive CD14 expression in differentiated HL60 cells was about 9 times higher than that of the control at the dose of 108 mol/L and 20 times higher than that of the control at the dose of 105 mol/L respectively, and this action involved upregulation of MafB gene in HL60 leukemia cells On the other hand, H89712 inhibited proliferation of K562 leukemia cells and the inhibition of K562 leukemia cells in MTT assay was shown by 34% at the dose of 108 mol/L and 88% at the dose of 105 mol/L respectively The inhibition of K562 leukemia cells in colony assay was 53% at the dose of 108 mol/L and 100% at the dose of 105 mol/L respectively H89712 also induced K562 cells toward erythroidlike differentiation and it was verified by flow cytometry that the percentage of positive CD71 expression in differentiated K562 cells was about 9 times higher than that of the control at the dose of 108 mol/L and 16 times higher than that of the control at the dose of 105 mol/L respectively This action was related to upregulation of Egr1 gene in K562 leukemia cellsConclusions Our results showed the important roles played by MafB in macrophage differentiation and Egr1 in erythroid differentiation of human myeloid leukemia cells
