目的制备膀胱癌特异性核基蛋白BLCA-4单克隆抗体。方法根据已确定的膀胱特癌异性核基蛋白BLCA-4的特异性氨基酸片段序列：羧基端－EISQLNAGAC－氨基端，进行多肽合成，将多肽用Imjeet Immtmogen EDC Conjugation Kit耦联KLH作为抗原，...目的制备膀胱癌特异性核基蛋白BLCA-4单克隆抗体。方法根据已确定的膀胱特癌异性核基蛋白BLCA-4的特异性氨基酸片段序列：羧基端－EISQLNAGAC－氨基端，进行多肽合成，将多肽用Imjeet Immtmogen EDC Conjugation Kit耦联KLH作为抗原，用杂交瘤细胞制备BLCA-4单克隆抗体结果得到纯化抗BLCA-4单克隆抗体3F10及2F11。3F10表达于膀胱尿路上皮癌及癌旁尿路上皮细胞核内，正常膀胱尿路上皮无表达。结论制备的膀胱癌特异性核基蛋白BLCA-4单克隆抗体初步结果显示有较高的敏感性和特异性。展开更多
目的:探讨非肌层浸润性膀胱癌患者血清和尿液膀胱癌特异性核基质蛋白-(1bladder cancer specific antigen-1,BLCA-1)含量的变化和意义。方法:选取22例非肌层浸润性膀胱癌患者作为实验组,23例前列腺增生患者作为前列腺增生组,21例健康体...目的:探讨非肌层浸润性膀胱癌患者血清和尿液膀胱癌特异性核基质蛋白-(1bladder cancer specific antigen-1,BLCA-1)含量的变化和意义。方法:选取22例非肌层浸润性膀胱癌患者作为实验组,23例前列腺增生患者作为前列腺增生组,21例健康体检者作为正常组。采用ELISA法检测所有受试者血清和尿液样本中BLCA-1的含量,并计算敏感度和特异度。结果:实验组患者尿液BLCA-1的含量明显高于前列腺增生组和正常组(P<0.05);尿液BLCA-1诊断非肌层浸润性膀胱癌的敏感度为59%(13/22),特异度为93%(41/44)。实验组和前列腺增生组患者血清BLCA-1含量明显高于正常组(P<0.05);血清BLCA-1诊断非肌层浸润性膀胱癌的敏感度为86%(19/22),特异度为56%(24/43)。结论:BLCA-1作为非肌层浸润性膀胱癌的新型检测指标,具有较高的敏感度和特异度,值得深入研究。展开更多
目的:探讨膀胱癌患者尿液膀胱特异性核基质蛋白(bladder cancer specific nuclear matrix proteins,BLCA)-1/-4水平及其临床应用价值。方法:本研究纳入38例膀胱癌患者、40例正常对照组。采集受试者尿液样本,通过竞争性酶联免疫吸附法(en...目的:探讨膀胱癌患者尿液膀胱特异性核基质蛋白(bladder cancer specific nuclear matrix proteins,BLCA)-1/-4水平及其临床应用价值。方法:本研究纳入38例膀胱癌患者、40例正常对照组。采集受试者尿液样本,通过竞争性酶联免疫吸附法(enzymelinked immunosorbent assay,ELISA)定量分析尿液中BLCA-1和BLCA-4的水平,绘制受试者工作曲线,确定cut-off值。结果:膀胱癌患者尿液BLCA-1/-4水平均显著高于对照组(P<0.001);当cut-off值取0.859 ng/mL时,BLCA-1诊断膀胱癌的敏感性和特异性分别为71%(27/38)、90%(36/40)。肌层浸润性膀胱癌患者尿液BLCA-1较非肌层浸润性膀胱癌患者水平显著升高(P<0.001),但不同分级膀胱癌患者尿液BLCA-4水平无显著差异(P>0.05)。高级别膀胱癌患者尿液BLCA-4水平较低级别膀胱癌患者显著升高(P<0.05),但不同分期膀胱癌患者尿液BLCA-4水平无显著差异(P>0.05)。以cut-off为0.859 ng/mL时,BLCA-1诊断膀胱癌的敏感性和特异性分别为71%(27/38)、90%(36/40)。以cut-off为0.620 ng/mL时,BLCA-4诊断膀胱癌的敏感性和特异性分别为76.3%(29/38)、97.5%(39/40)。联合检测尿液BLCA-1和BLCA-4诊断膀胱癌的敏感性和特异性分别为84.2%(32/38)和100%(40/40),准确度为0.923(77/78),阳性预测值为100%(32/32),阴性预测值为86.9%(40/46)以及YOUDEN指数分别为0.842。结论:膀胱癌患者尿液BLCA-1和BLCA-4水平显著升高,且敏感性和特异性均较高。联合检测尿液BLCA-1和BLCA-4较单一检测用于诊断膀胱癌的临床应用价值更高。展开更多
Background:The treatment alternatives for bladder cancer(BLCA),the 10th most prevalent cancer in the world,need to be further investigated,and many active substances like Puerarin in herbal medicine were found to be e...Background:The treatment alternatives for bladder cancer(BLCA),the 10th most prevalent cancer in the world,need to be further investigated,and many active substances like Puerarin in herbal medicine were found to be effective in treating BLCA.The purpose of this study was to investigate the potential treating mechanisms of Puerarin on BLCA.Methods:The cell counting kit 8 assay and flow cytometry were performed to confirm Puerarin’s ability to suppress BLCA.The differentially expressed proteins(DEPs)were obtained by Tandem Mass Tags technology and functional enrichment analysis performed by R studio.The most enriched pathways were selected for study and the DEPs were screened out.Protein-protein interaction network maps were created using String and Cytoscape and key proteins,which will be analyzed for survival,expression,and upstream transcription factor prediction,were screened out using the cytoHubba plugin.CHEA3 was used to obtain upstream transcription factor validated by molecular docking and western blotting experiments.Results:Cell counting kit 8 showed that Puerarin inhibited BLCA cells,with 50%inhibitory concentration of 218μmol/L in T24 and 198μmol/L in 5637.Flow cytometry reveals that Puerarin blocks T24 and 5637 cells in G1 phase.1,385 DEPs were obtained and the enrichment analysis revealed that cell cycle and DNA replication were the two main areas in which DEPs were enriched.Cyclin-B-cyclin dependent kinase 1(CDK1),cyclin B1(CCNB1),and polo-like kinase 1(PLK1)were identified as key proteins,and their upstream transcription factor was predicted to be centromere protein A(CENPA).Puerarin’s binding energy to CENPA was determined by molecular docking to be−6.3 kcal/mol,indicating a strong binding interaction.Western blot showed that Puerarin significantly reduced the expression of CENPA.Conclusion:We hypothesize that Puerarin may inhibit the proliferation of bladder cancer cells by inhibiting CENPA expression to regulate PLK1 and CCNB1 expression,thereby affecting cell cycle.展开更多
文摘目的制备膀胱癌特异性核基蛋白BLCA-4单克隆抗体。方法根据已确定的膀胱特癌异性核基蛋白BLCA-4的特异性氨基酸片段序列：羧基端－EISQLNAGAC－氨基端，进行多肽合成，将多肽用Imjeet Immtmogen EDC Conjugation Kit耦联KLH作为抗原，用杂交瘤细胞制备BLCA-4单克隆抗体结果得到纯化抗BLCA-4单克隆抗体3F10及2F11。3F10表达于膀胱尿路上皮癌及癌旁尿路上皮细胞核内，正常膀胱尿路上皮无表达。结论制备的膀胱癌特异性核基蛋白BLCA-4单克隆抗体初步结果显示有较高的敏感性和特异性。
文摘目的:探讨非肌层浸润性膀胱癌患者血清和尿液膀胱癌特异性核基质蛋白-(1bladder cancer specific antigen-1,BLCA-1)含量的变化和意义。方法:选取22例非肌层浸润性膀胱癌患者作为实验组,23例前列腺增生患者作为前列腺增生组,21例健康体检者作为正常组。采用ELISA法检测所有受试者血清和尿液样本中BLCA-1的含量,并计算敏感度和特异度。结果:实验组患者尿液BLCA-1的含量明显高于前列腺增生组和正常组(P<0.05);尿液BLCA-1诊断非肌层浸润性膀胱癌的敏感度为59%(13/22),特异度为93%(41/44)。实验组和前列腺增生组患者血清BLCA-1含量明显高于正常组(P<0.05);血清BLCA-1诊断非肌层浸润性膀胱癌的敏感度为86%(19/22),特异度为56%(24/43)。结论:BLCA-1作为非肌层浸润性膀胱癌的新型检测指标,具有较高的敏感度和特异度,值得深入研究。
基金supported by National Natural Science Fund Item Number(82004110)Xuzhou Science and Technology Plan Project(KC22096)+3 种基金China Postdoctoral Science Foundation(2022M722674)Xuzhou Medical Reserve Talents Project(XWRCHT20220009)the Xuzhou Clinical Medicine Expert Team Project(2018TD004)Peixian Science and Technology Program(P202410)。
文摘Background:The treatment alternatives for bladder cancer(BLCA),the 10th most prevalent cancer in the world,need to be further investigated,and many active substances like Puerarin in herbal medicine were found to be effective in treating BLCA.The purpose of this study was to investigate the potential treating mechanisms of Puerarin on BLCA.Methods:The cell counting kit 8 assay and flow cytometry were performed to confirm Puerarin’s ability to suppress BLCA.The differentially expressed proteins(DEPs)were obtained by Tandem Mass Tags technology and functional enrichment analysis performed by R studio.The most enriched pathways were selected for study and the DEPs were screened out.Protein-protein interaction network maps were created using String and Cytoscape and key proteins,which will be analyzed for survival,expression,and upstream transcription factor prediction,were screened out using the cytoHubba plugin.CHEA3 was used to obtain upstream transcription factor validated by molecular docking and western blotting experiments.Results:Cell counting kit 8 showed that Puerarin inhibited BLCA cells,with 50%inhibitory concentration of 218μmol/L in T24 and 198μmol/L in 5637.Flow cytometry reveals that Puerarin blocks T24 and 5637 cells in G1 phase.1,385 DEPs were obtained and the enrichment analysis revealed that cell cycle and DNA replication were the two main areas in which DEPs were enriched.Cyclin-B-cyclin dependent kinase 1(CDK1),cyclin B1(CCNB1),and polo-like kinase 1(PLK1)were identified as key proteins,and their upstream transcription factor was predicted to be centromere protein A(CENPA).Puerarin’s binding energy to CENPA was determined by molecular docking to be−6.3 kcal/mol,indicating a strong binding interaction.Western blot showed that Puerarin significantly reduced the expression of CENPA.Conclusion:We hypothesize that Puerarin may inhibit the proliferation of bladder cancer cells by inhibiting CENPA expression to regulate PLK1 and CCNB1 expression,thereby affecting cell cycle.
