BACKGROUND Hepatitis B virus(HBV)infection is a leading cause of global hepatocellular carcinoma(HCC).Conventional biomarkers such as alpha-fetoprotein(AFP)demonstrate suboptimal sensitivity and specificity.Emerging e...BACKGROUND Hepatitis B virus(HBV)infection is a leading cause of global hepatocellular carcinoma(HCC).Conventional biomarkers such as alpha-fetoprotein(AFP)demonstrate suboptimal sensitivity and specificity.Emerging evidence suggests that serum extra spindle pole bodies like 1(ESPL1)protein and p53 antibody may improve diagnostic accuracy.AIM To assess and compare the diagnostic performance of serum ESPL1 protein and p53 antibody in HBV-related HCC(HBV-HCC).METHODS This case-control study from the First Affiliated Hospital of Guangxi Medical University enrolled 30 patients with chronic hepatitis B(CHB),30 with HBV-related liver cirrhosis(HBV-LC),55 with HBV-HCC,and 30 healthy controls.Serum ESPL1 protein and p53 antibody levels were quantified via ELISA.Diagnostic performance was evaluated using receiver operating characteristic(ROC)curve analysis,including sensitivity,specificity,and correlation with AFP.RESULTS Serum ESPL1 protein levels progressively increased across disease stages(CHB:89.9 ng/L;HBV-LC:188.83 ng/L;HBV-HCC:317.63 ng/L),with a significantly higher area under the ROC curve(AUC=0.917)than either p53 antibody(AUC=0.725)or AFP(AUC=0.678).p53 antibody levels were significantly elevated only in the HBVHCC group.ESPL1 demonstrated superior sensitivity and concordance with histopathological findings.A significant correlation between ESPL1 and p53 antibody levels was observed exclusively in the HBV-HCC group(r=0.320,P=0.017),suggesting potential interplay in malignant transformation.CONCLUSION Serum ESPL1 protein,a promising biomarker for early HBV-HCC detection,outperforms p53 antibody in diagnostic reliability.Higher ESPL1 levels correlate with increased HCC risk in chronic HBV patients.展开更多
报告以脑积水起病的晚发型戊二酸血症1型(glutaric acidemia type 1,GA1)1例。患者女性,21岁,以急性脑积水为主要临床表现,头颅MRI示脑内大范围脑白质、双侧基底节区及小脑半球蚓部异常信号,双侧脑室扩张,双侧颞极蛛网膜囊肿。血尿有机...报告以脑积水起病的晚发型戊二酸血症1型(glutaric acidemia type 1,GA1)1例。患者女性,21岁,以急性脑积水为主要临床表现,头颅MRI示脑内大范围脑白质、双侧基底节区及小脑半球蚓部异常信号,双侧脑室扩张,双侧颞极蛛网膜囊肿。血尿有机酸分析检出大量戊二酸和3-羟基戊二酸,GCDH基因检测发现为复合杂合性突变(S119L和R355H),确诊为戊二酸血症1型,给予相关治疗后,症状缓解。可见临床上如遇上难以解释的脑积水,头部MRI示典型对称性病灶,包括小脑损害,应进行血尿有机酸筛查以除外代谢性疾病。展开更多
目的:构建包含突变型CDK4基因的真核绿色荧光表达载体,作为POLD1基因依赖的细胞周期复制调控的模型,研究POLD1基因相关的癌性增殖的机制,为干预细胞恶性增殖提供新的思路.方法:设计人突变型CDK4基因全长特异性引物进行P C R扩增人肝癌...目的:构建包含突变型CDK4基因的真核绿色荧光表达载体,作为POLD1基因依赖的细胞周期复制调控的模型,研究POLD1基因相关的癌性增殖的机制,为干预细胞恶性增殖提供新的思路.方法:设计人突变型CDK4基因全长特异性引物进行P C R扩增人肝癌细胞系SMMC-7721总cDNA,以pEGFP-C1质粒为模板连接,得到重组质粒GFP-CDK4后进行测序和生物信息学比对分析;转染细胞分3组:实验组(转染突变型CDK4重组真核表达质粒GFP-CDK4),阴性对照组(转染空载体pEGFP-C1组)和空白对照组(SMMC-7702).通过MTT试验分析细胞增殖变化;实时荧光定量PCR技术检测CDK4、POLD1及细胞周期相关因子的表达量,Western blot检测蛋白表达的差异.结果:成功构建了人突变型CDK4基因真核表达质粒GFP-CDK4,转染到肝细胞SMMC-7702后使细胞表达融合绿色荧光的C D K4蛋白;S M M C-7721细胞中突变型的C D K4存在5个碱基突变,4个碱基插入,2个碱基缺失,这使得5个氨基酸序列发生了改变;与空白对照组及阴性对照组相比,实验组细胞增殖明显升高(0.826±0.08vs0.596±0.06,0.609±0.10,F=7.033,均P<0.05);实验组CDK4mRNA表达水平差异明显(1.94±0.11vs1.01±0.00,1.05±0.12,F=54.046,P<0.01),POLD1mRNA相应地升高(2.47±0.25vs1.16±0.00,1.26±0.23,F=135.496,P<0.01);稳定转染细胞的蛋白水平变化趋势与基因相同,其中实验组CDK4(0.65±0.03vs0.41±0.03,0.39±0.05,F=14.665,均P<0.05),P125(0.54±0.04vs0.30±0.07,0.25±0.06,F=11.788,均P<0.05).结论:人突变型CDK4基因的真核表达载体GFP-CDK4显著促进肝细胞的增殖能力,这与POLD1基因及P125蛋白的高表达相关.展开更多
基金Supported by National Natural Science Foundation of China,No.81960115,No.82160123 and No.82260124Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor(Guangxi Medical University),Ministry of Education,No.GKEZZ202107+1 种基金Guangxi Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor,No.GKE-ZZ202218Guangxi Science and Technology Program,No.AD25069077.
文摘BACKGROUND Hepatitis B virus(HBV)infection is a leading cause of global hepatocellular carcinoma(HCC).Conventional biomarkers such as alpha-fetoprotein(AFP)demonstrate suboptimal sensitivity and specificity.Emerging evidence suggests that serum extra spindle pole bodies like 1(ESPL1)protein and p53 antibody may improve diagnostic accuracy.AIM To assess and compare the diagnostic performance of serum ESPL1 protein and p53 antibody in HBV-related HCC(HBV-HCC).METHODS This case-control study from the First Affiliated Hospital of Guangxi Medical University enrolled 30 patients with chronic hepatitis B(CHB),30 with HBV-related liver cirrhosis(HBV-LC),55 with HBV-HCC,and 30 healthy controls.Serum ESPL1 protein and p53 antibody levels were quantified via ELISA.Diagnostic performance was evaluated using receiver operating characteristic(ROC)curve analysis,including sensitivity,specificity,and correlation with AFP.RESULTS Serum ESPL1 protein levels progressively increased across disease stages(CHB:89.9 ng/L;HBV-LC:188.83 ng/L;HBV-HCC:317.63 ng/L),with a significantly higher area under the ROC curve(AUC=0.917)than either p53 antibody(AUC=0.725)or AFP(AUC=0.678).p53 antibody levels were significantly elevated only in the HBVHCC group.ESPL1 demonstrated superior sensitivity and concordance with histopathological findings.A significant correlation between ESPL1 and p53 antibody levels was observed exclusively in the HBV-HCC group(r=0.320,P=0.017),suggesting potential interplay in malignant transformation.CONCLUSION Serum ESPL1 protein,a promising biomarker for early HBV-HCC detection,outperforms p53 antibody in diagnostic reliability.Higher ESPL1 levels correlate with increased HCC risk in chronic HBV patients.
文摘报告以脑积水起病的晚发型戊二酸血症1型(glutaric acidemia type 1,GA1)1例。患者女性,21岁,以急性脑积水为主要临床表现,头颅MRI示脑内大范围脑白质、双侧基底节区及小脑半球蚓部异常信号,双侧脑室扩张,双侧颞极蛛网膜囊肿。血尿有机酸分析检出大量戊二酸和3-羟基戊二酸,GCDH基因检测发现为复合杂合性突变(S119L和R355H),确诊为戊二酸血症1型,给予相关治疗后,症状缓解。可见临床上如遇上难以解释的脑积水,头部MRI示典型对称性病灶,包括小脑损害,应进行血尿有机酸筛查以除外代谢性疾病。
文摘目的:构建包含突变型CDK4基因的真核绿色荧光表达载体,作为POLD1基因依赖的细胞周期复制调控的模型,研究POLD1基因相关的癌性增殖的机制,为干预细胞恶性增殖提供新的思路.方法:设计人突变型CDK4基因全长特异性引物进行P C R扩增人肝癌细胞系SMMC-7721总cDNA,以pEGFP-C1质粒为模板连接,得到重组质粒GFP-CDK4后进行测序和生物信息学比对分析;转染细胞分3组:实验组(转染突变型CDK4重组真核表达质粒GFP-CDK4),阴性对照组(转染空载体pEGFP-C1组)和空白对照组(SMMC-7702).通过MTT试验分析细胞增殖变化;实时荧光定量PCR技术检测CDK4、POLD1及细胞周期相关因子的表达量,Western blot检测蛋白表达的差异.结果:成功构建了人突变型CDK4基因真核表达质粒GFP-CDK4,转染到肝细胞SMMC-7702后使细胞表达融合绿色荧光的C D K4蛋白;S M M C-7721细胞中突变型的C D K4存在5个碱基突变,4个碱基插入,2个碱基缺失,这使得5个氨基酸序列发生了改变;与空白对照组及阴性对照组相比,实验组细胞增殖明显升高(0.826±0.08vs0.596±0.06,0.609±0.10,F=7.033,均P<0.05);实验组CDK4mRNA表达水平差异明显(1.94±0.11vs1.01±0.00,1.05±0.12,F=54.046,P<0.01),POLD1mRNA相应地升高(2.47±0.25vs1.16±0.00,1.26±0.23,F=135.496,P<0.01);稳定转染细胞的蛋白水平变化趋势与基因相同,其中实验组CDK4(0.65±0.03vs0.41±0.03,0.39±0.05,F=14.665,均P<0.05),P125(0.54±0.04vs0.30±0.07,0.25±0.06,F=11.788,均P<0.05).结论:人突变型CDK4基因的真核表达载体GFP-CDK4显著促进肝细胞的增殖能力,这与POLD1基因及P125蛋白的高表达相关.
