为深入研究山羊AP1M1(adaptor related protein complex 1 subunit mu 1)在机体生长和繁殖中的作用,采用RT-PCR方法克隆了山羊AP1M1基因,并对其序列和编码蛋白进行分析;构建了pET-30α-AP1M1原核表达载体,免疫实验兔,制备AP1M1基因多克...为深入研究山羊AP1M1(adaptor related protein complex 1 subunit mu 1)在机体生长和繁殖中的作用,采用RT-PCR方法克隆了山羊AP1M1基因,并对其序列和编码蛋白进行分析;构建了pET-30α-AP1M1原核表达载体,免疫实验兔,制备AP1M1基因多克隆抗体;采用qPCR和Western blot检测比较AP1M1在山羊不同组织及睾丸不同发育时期的表达特征。结果显示,山羊AP1M1基因CDS为1 272 bp,编码423个氨基酸,具有1个Clat-adaptor结构域、1个Adap-comp结构域和5个蛋白质-蛋白质结合位点,在哺乳动物中极为保守。间接ELISA和Western blot结果表明,制备的兔抗pET-30α-AP1M1多克隆抗体效价为1∶128 000,可以与重组蛋白特异性结合。表达模式检测结果显示,AP1M1在成年山羊各组织中均有不同程度的表达,其中在肾脏的表达量最高,其次是睾丸,均极显著高于其他组织(P<0.01);此外,AP1M1在山羊不同发育时期睾丸组织中均有表达,但在性成熟时显著增加(P<0.01),体成熟时达到最高。本试验为进一步探索该基因在山羊睾丸中的功能奠定基础。展开更多
Objective:The World Health Organization(WHO)grading based on histopathology cannot always accurately predict tumor behavior of meningiomas.To overcome the limitations of the WHO grading,the study aims to propose a nov...Objective:The World Health Organization(WHO)grading based on histopathology cannot always accurately predict tumor behavior of meningiomas.To overcome the limitations of the WHO grading,the study aims to propose a novel oxidative stress-based molecular classification for WHO grade 2/3 meningiomas.Methods:Differentially expressed oxidative stress-related genes were analyzed between 86 WHO grade 1(low grade)meningiomas and 99 grade 2/3(high grade)meningiomas.An oxidative stress-based molecular classification was developed in high-grade meningiomas through consensus clustering analysis.Immune microenvironment features,responses to immunotherapy and chemotherapy,and targeted drugs were evaluated.Three machine learning models:logistic regression,support vector machine,and random forest,were built for differentiating the classification.Key oxidative stress-related geneswere verified in humanmeningeal cells(HMC)and two meningioma cells(CH-157MN and IOMMLee)via reverse transcription quantitative polymerase chain reaction(RT-qPCR)and western blot.After knockdown of Forkhead Box M1(FOXM1)or Prion Protein(PRNP),cell growth,migration,and reactive oxygen species(ROS)levels were measured through cell counting kit-8(CCK-8),transwell,and immunofluorescence,respectively.Results:We classified high-grade meningiomas into two oxidative stress-based clusters,termed cluster 1 and cluster 2.Cluster 1 exhibited higher infiltrations of immune and stromal cells and higher expression of classic immune checkpoints:Cluster of Differentiation 86(CD86),Programmed Cell Death 1(PDCD1),and Leukocyte-Associated Immunoglobulin-Like Receptor 1(LAIR1),indicating that cluster 1 meningiomas might respond to immunotherapy.Drug sensitivity was heterogeneous between the two clusters.Three classifiers were established,which could accurately differentiate this molecular classification.FOXM1 and PRNP were experimentally evidenced to be highly expressed inmeningioma cells,and their knockdown hindered cell growth and migration and triggered ROS accumulation.Conclusion:In summary,our findings established a novel oxidative stress-based molecular classification and identified potential treatment vulnerabilities in high-grade meningiomas,which might assist personalized clinical management.展开更多
对丙烯二聚生产4-甲基-1-戊烯(4M1P)的反应机理进行文献研究,根据文献值计算反应器出口物料的理论组成,作为分离部分的进料组成。对丙烯二聚反应的产品分离部分流程进行设计,运用Aspen Plus 软件对产品分离流程进行流程模拟计算。在此...对丙烯二聚生产4-甲基-1-戊烯(4M1P)的反应机理进行文献研究,根据文献值计算反应器出口物料的理论组成,作为分离部分的进料组成。对丙烯二聚反应的产品分离部分流程进行设计,运用Aspen Plus 软件对产品分离流程进行流程模拟计算。在此基础上进一步优化热交换系统和优化塔的操作条件等方式,计算出各精馏塔器的最佳理论板数、操作温度、压力等数据,为丙烯二聚生产4-甲基-1-戊烯的国产化技术开发提供参考。展开更多
FOXM1(Forkhead box protein M1)是调控细胞增殖的重要转录因子,近年来研究表明与肿瘤发生密切相关,但与卵巢癌的关系尚不明确.通过检测68例卵巢癌标本、21例卵巢良性肿瘤标本、24例正常卵巢标本以及3株卵巢癌细胞系(A2780细胞、OVCAR3...FOXM1(Forkhead box protein M1)是调控细胞增殖的重要转录因子,近年来研究表明与肿瘤发生密切相关,但与卵巢癌的关系尚不明确.通过检测68例卵巢癌标本、21例卵巢良性肿瘤标本、24例正常卵巢标本以及3株卵巢癌细胞系(A2780细胞、OVCAR3细胞、SKOV3细胞)中FOXM1的表达情况,分析其与临床参数之间的相关性及临床意义.结果显示卵巢癌中FOXM1表达明显高于卵巢良性肿瘤及正常卵巢组织,差异极为显著,其中低分化细胞中表达强于中高分化细胞(P=0.013),Ⅲ～Ⅳ期表达强于Ⅰ～Ⅱ期(P=0.011),但与病理类型无关;FOXM1在3株卵巢癌细胞系中均有较强表达.FOXM1在卵巢癌组织及3种卵巢癌细胞系中存在高表达,且与卵巢癌分化程度及临床分期有关.展开更多
文摘为深入研究山羊AP1M1(adaptor related protein complex 1 subunit mu 1)在机体生长和繁殖中的作用,采用RT-PCR方法克隆了山羊AP1M1基因,并对其序列和编码蛋白进行分析;构建了pET-30α-AP1M1原核表达载体,免疫实验兔,制备AP1M1基因多克隆抗体;采用qPCR和Western blot检测比较AP1M1在山羊不同组织及睾丸不同发育时期的表达特征。结果显示,山羊AP1M1基因CDS为1 272 bp,编码423个氨基酸,具有1个Clat-adaptor结构域、1个Adap-comp结构域和5个蛋白质-蛋白质结合位点,在哺乳动物中极为保守。间接ELISA和Western blot结果表明,制备的兔抗pET-30α-AP1M1多克隆抗体效价为1∶128 000,可以与重组蛋白特异性结合。表达模式检测结果显示,AP1M1在成年山羊各组织中均有不同程度的表达,其中在肾脏的表达量最高,其次是睾丸,均极显著高于其他组织(P<0.01);此外,AP1M1在山羊不同发育时期睾丸组织中均有表达,但在性成熟时显著增加(P<0.01),体成熟时达到最高。本试验为进一步探索该基因在山羊睾丸中的功能奠定基础。
基金supported by Hubei Provincial Natural Science Foundation of China(grants 2023AFB208)the Chinese Primary Health Care Foundation(Grant No.cphcf-2022-222)+2 种基金2025 Hubei Provincial Natural Science Foundation Innovation and Development Joint Fund Project:(JCZRLH202500457)Shanghai Foundation for Anti-Cancer&Cancer Prevention Development Phase II Exploration Oncology Research Fund Project:“Study on the Mechanism of ANO9-Mediated Cetuximab Resistance in Head and Neck Squamous Cell Carcinoma”(No Grant Number)Qingdao Sheci Public Welfare Relief Center Pan-Cancer Treatment Research Fund Project:(QD-HN30008).
文摘Objective:The World Health Organization(WHO)grading based on histopathology cannot always accurately predict tumor behavior of meningiomas.To overcome the limitations of the WHO grading,the study aims to propose a novel oxidative stress-based molecular classification for WHO grade 2/3 meningiomas.Methods:Differentially expressed oxidative stress-related genes were analyzed between 86 WHO grade 1(low grade)meningiomas and 99 grade 2/3(high grade)meningiomas.An oxidative stress-based molecular classification was developed in high-grade meningiomas through consensus clustering analysis.Immune microenvironment features,responses to immunotherapy and chemotherapy,and targeted drugs were evaluated.Three machine learning models:logistic regression,support vector machine,and random forest,were built for differentiating the classification.Key oxidative stress-related geneswere verified in humanmeningeal cells(HMC)and two meningioma cells(CH-157MN and IOMMLee)via reverse transcription quantitative polymerase chain reaction(RT-qPCR)and western blot.After knockdown of Forkhead Box M1(FOXM1)or Prion Protein(PRNP),cell growth,migration,and reactive oxygen species(ROS)levels were measured through cell counting kit-8(CCK-8),transwell,and immunofluorescence,respectively.Results:We classified high-grade meningiomas into two oxidative stress-based clusters,termed cluster 1 and cluster 2.Cluster 1 exhibited higher infiltrations of immune and stromal cells and higher expression of classic immune checkpoints:Cluster of Differentiation 86(CD86),Programmed Cell Death 1(PDCD1),and Leukocyte-Associated Immunoglobulin-Like Receptor 1(LAIR1),indicating that cluster 1 meningiomas might respond to immunotherapy.Drug sensitivity was heterogeneous between the two clusters.Three classifiers were established,which could accurately differentiate this molecular classification.FOXM1 and PRNP were experimentally evidenced to be highly expressed inmeningioma cells,and their knockdown hindered cell growth and migration and triggered ROS accumulation.Conclusion:In summary,our findings established a novel oxidative stress-based molecular classification and identified potential treatment vulnerabilities in high-grade meningiomas,which might assist personalized clinical management.
文摘对丙烯二聚生产4-甲基-1-戊烯(4M1P)的反应机理进行文献研究,根据文献值计算反应器出口物料的理论组成,作为分离部分的进料组成。对丙烯二聚反应的产品分离部分流程进行设计,运用Aspen Plus 软件对产品分离流程进行流程模拟计算。在此基础上进一步优化热交换系统和优化塔的操作条件等方式,计算出各精馏塔器的最佳理论板数、操作温度、压力等数据,为丙烯二聚生产4-甲基-1-戊烯的国产化技术开发提供参考。
文摘FOXM1(Forkhead box protein M1)是调控细胞增殖的重要转录因子,近年来研究表明与肿瘤发生密切相关,但与卵巢癌的关系尚不明确.通过检测68例卵巢癌标本、21例卵巢良性肿瘤标本、24例正常卵巢标本以及3株卵巢癌细胞系(A2780细胞、OVCAR3细胞、SKOV3细胞)中FOXM1的表达情况,分析其与临床参数之间的相关性及临床意义.结果显示卵巢癌中FOXM1表达明显高于卵巢良性肿瘤及正常卵巢组织,差异极为显著,其中低分化细胞中表达强于中高分化细胞(P=0.013),Ⅲ～Ⅳ期表达强于Ⅰ～Ⅱ期(P=0.011),但与病理类型无关;FOXM1在3株卵巢癌细胞系中均有较强表达.FOXM1在卵巢癌组织及3种卵巢癌细胞系中存在高表达,且与卵巢癌分化程度及临床分期有关.
