目的探究Cosmc或T-合酶基因敲除引起的异常O-糖基化修饰对结肠癌外泌体中微小RNA表达模式的影响,以揭示O-糖基化在结肠癌发展中的分子机制,并识别潜在的生物标志物,为结肠癌的早期诊断和治疗提供新策略。方法在人结肠癌细胞株HCT116中应...目的探究Cosmc或T-合酶基因敲除引起的异常O-糖基化修饰对结肠癌外泌体中微小RNA表达模式的影响,以揭示O-糖基化在结肠癌发展中的分子机制,并识别潜在的生物标志物,为结肠癌的早期诊断和治疗提供新策略。方法在人结肠癌细胞株HCT116中应用CRISPR/Cas-9基因编辑技术,分别靶向敲除Cosmc或T-合酶基因,以构建两种异常O-糖基化修饰的稳定转染细胞系。提取分离出肠癌细胞来源的外泌体,利用微小RNA(microRNAs,miRNAs)微阵列芯片,对比分析了其外泌体中miRNAs的表达差异。随后,在两组细胞来源的外泌体中筛选出表达量发生显著变化且变化趋势一致的miRNAs群体,采用传统荧光定量聚合酶链式反应(polymerase chain reaction,PCR)技术对这些miRNAs进行了独立验证。最后,通过癌症基因组图谱计划(The Cancer Genome Atlas Program,TCGA)数据库获取结直肠癌患者信息,利用R语言将表达上调的miRNA进行基因集富集分析(Gene Set Enrichment Analysis,GSEA),探索其显著相关的下游通路及表型。结果无论是Cosmc还是T-合酶基因缺失均会导致结肠癌细胞中O-糖基化修饰发生异常,Tn抗原暴露,进而使得结肠癌细胞来源的外泌体中hsa-miR-125b-1-3p表达量下调(P<0.05),而hsa-miR-218-5p表达量上调(P<0.05),且与肿瘤细胞上皮-间质转化过程密切相关(P<0.05)。结论由Cosmc或T-合酶基因缺失所介导的异常O-糖基化对结肠癌外泌体中相关miRNAs表达产生显著影响,进而可能影响结肠癌上皮-间质转化过程,从而促进肠癌远处转移。由此,基于肠癌外泌体的高稳定性及易检出性的天然优势,通过对其所携带miRNAs的表达量变化进行检测,可监测疾病进展和治疗效果,从而为结肠癌患者提供更为个性化的诊疗策略。展开更多
Background:Terpinen-4-ol(T4O),a key constituent of tea tree essential oil and various aromatic plants,has shown promising antiproliferative and pro-apoptotic effects in melanoma and other cancer types.However,its effi...Background:Terpinen-4-ol(T4O),a key constituent of tea tree essential oil and various aromatic plants,has shown promising antiproliferative and pro-apoptotic effects in melanoma and other cancer types.However,its efficacy against cutaneous squamous cell carcinoma(cSCC)remains unclear.Thus,in this study,we investigated the in vivo and in vitro effects of T4O on cSCC cell lines and preliminarily explored its impacting pathways.Methods:Using CCK8 and assay colony formation,we assessed the viability of cSCC A431,SCL-1,and COLO-16 cells treated with T40 at varying concentrations(0,1,2,and 4μM).Flow cytometry was employed to evaluate T4O’s effect on cSCC cell’s cycle progression and apoptosis induction.Additionally,western blotting was utilized to examine the expression intensities of N-cadherin and E-cadherin,two indicative markers of the epithelial-mesenchymal transition(EMT)pathway.T4O’s in vivo effect on inhibiting tumor progression was evaluated on an established xenograft tumor model.Then,the molecular mechanisms of T4O’s antitumor effect were explored by an integrated genome-wide transcriptomics and proteomics study on cSCC A431c cells.Finally,calpain-2’s potential mediator role in T4O’s anti-tumor mechanism was investigated in calpain-2 knockdown cell lines prepared via siRNA transfection.Result:It’s demonstrated that T4O treatment inhibited cSCC proliferation,clonogenicity,migration,and invasion while inducing apoptosis and suppressing the EMT pathway.T4O administration also inhibited cSCC tumorigenesis in the xenograft tumor model.RNA-sequencing and iTRAQ analysis detected significant upregulation of calpain-2 expression in T4O-treated cSCC cells.Western blotting confirmed that T4O significantly increased calpain-2 expression and promoted proteolytic cleavage ofβ-catenin and caspase-12,two calpain-2 target proteins.Importantly,siRNA-mediated calpain-2 knockdown relieved T4O’s suppressive effect on cSCC cell proliferation and motility.Mechanistically,T4O upregulates calpain-2 expression and promotes the cleavage ofβ-catenin and caspase-12,with siRNA-mediated calpain-2 knockdown mitigating T4O’s suppressive effects.Conclusion:These findings suggest that T4O’s antitumor activity in cSCC is mediated through the upregulation of calpain-2 expression and subsequent modulation ofβ-catenin and caspase-12.展开更多
End-stage kidney failure(ESKD)is a global issue where kidney replacement therapy imposes enormous economic burden to people of developing countries,in addition to the severe limitations to the availability of hemodial...End-stage kidney failure(ESKD)is a global issue where kidney replacement therapy imposes enormous economic burden to people of developing countries,in addition to the severe limitations to the availability of hemodialysis and peritoneal dialysis technique.The best option of kidney transplantation also requires lifelong combination immunosuppressive medicines,the cost of which is equally comparable to lifelong dialysis.A strategy of achieving transplant tolerance that requires minimum immunosuppressive medicines,although in experimental stage,also requires state-of-art technology with costly medicines and interventions.This is evidently beyond the reach of ESKD patients of developing countries.Hence,globally in developing countries,a need for an innovative but cost-effective tolerance protocol is a burning need for a successful transplant program.In brief,transplant tolerance is defined as a state of donorspecific unresponsiveness to the allograft antigens without the need for ongoing pharmacologic immunosuppression or with a minimal need.Current state-of-art techniques involves:(1)A state of hematological chimera,for complete tolerance;(2)Prope or partial tolerance where immune-reactive T-lymphocytes are inhibited using monoclonal antibodies;and(3)Chimeric antigen receptor for T-regulatory(T-reg)cell therapy using genetically engineered T-reg cells targeting specific Tlymphocyte receptors for inducing anergy.From our real-world experience in transplant management in post-transplant lympho-proliferative disorders(PTLD),we noticed frequently a drastic reduction in the need of immunosuppressive medicines following lympho-ablative therapy for PTLD.We recently published a case study on a real-world experience transplant case where we explained a partial or prope tolerance that developed after lymphocyte ablation therapy,following which the allograft was maintained with low dose dual standard immunosuppressive medicines.Based on this publication,we propose here an innovative tolerance protocol for living related low risk kidney transplantation for developing countries,in this opinion review.展开更多
通过对100 t LF精炼37Mn5套管钢时1 540～1580℃钢水中稳定氧化物夹杂含量与T[O]的关系以及钢-渣平衡时渣碱度对钢中活性氧含量影响的分析,提出降低钢中T[O]和夹杂物含量的改进工艺措施。热力学计算和试验结果表明,当[Al]s为0.03%～0.0...通过对100 t LF精炼37Mn5套管钢时1 540～1580℃钢水中稳定氧化物夹杂含量与T[O]的关系以及钢-渣平衡时渣碱度对钢中活性氧含量影响的分析,提出降低钢中T[O]和夹杂物含量的改进工艺措施。热力学计算和试验结果表明,当[Al]s为0.03%～0.04%,精炼渣碱度为3,(CaO)/(Al_2O_3)=3～3.5时,37Mn5钢中的氧含量降到10×10^(-6),管材基本消除了粗系夹杂,细系夹杂A+B+C+D≤5.0级,达到使用要求。展开更多
T2O(Television to Online)是互联网+时代一种从电视到电商的新型营销传播模式。本研究结合创新扩散理论、SOR模型、计划行为理论、TAM模型等传播学、心理学、消费者行为学的跨学科理论,通过深度访谈、问卷调查等方式获取T2O营销传播模...T2O(Television to Online)是互联网+时代一种从电视到电商的新型营销传播模式。本研究结合创新扩散理论、SOR模型、计划行为理论、TAM模型等传播学、心理学、消费者行为学的跨学科理论,通过深度访谈、问卷调查等方式获取T2O营销传播模式下影响消费者行为意向的因素。发现在T2O模式下,消费者的感知态度、主观规范和个人创新性都会对其行为意向产生显著影响,其中感知娱乐的影响最大。本文为T2O的研究提供了新的角度,同时为电视台、电商和广告主深入实践应用T2O模式提出了建议。展开更多
文摘目的探究Cosmc或T-合酶基因敲除引起的异常O-糖基化修饰对结肠癌外泌体中微小RNA表达模式的影响,以揭示O-糖基化在结肠癌发展中的分子机制,并识别潜在的生物标志物,为结肠癌的早期诊断和治疗提供新策略。方法在人结肠癌细胞株HCT116中应用CRISPR/Cas-9基因编辑技术,分别靶向敲除Cosmc或T-合酶基因,以构建两种异常O-糖基化修饰的稳定转染细胞系。提取分离出肠癌细胞来源的外泌体,利用微小RNA(microRNAs,miRNAs)微阵列芯片,对比分析了其外泌体中miRNAs的表达差异。随后,在两组细胞来源的外泌体中筛选出表达量发生显著变化且变化趋势一致的miRNAs群体,采用传统荧光定量聚合酶链式反应(polymerase chain reaction,PCR)技术对这些miRNAs进行了独立验证。最后,通过癌症基因组图谱计划(The Cancer Genome Atlas Program,TCGA)数据库获取结直肠癌患者信息,利用R语言将表达上调的miRNA进行基因集富集分析(Gene Set Enrichment Analysis,GSEA),探索其显著相关的下游通路及表型。结果无论是Cosmc还是T-合酶基因缺失均会导致结肠癌细胞中O-糖基化修饰发生异常,Tn抗原暴露,进而使得结肠癌细胞来源的外泌体中hsa-miR-125b-1-3p表达量下调(P<0.05),而hsa-miR-218-5p表达量上调(P<0.05),且与肿瘤细胞上皮-间质转化过程密切相关(P<0.05)。结论由Cosmc或T-合酶基因缺失所介导的异常O-糖基化对结肠癌外泌体中相关miRNAs表达产生显著影响,进而可能影响结肠癌上皮-间质转化过程,从而促进肠癌远处转移。由此,基于肠癌外泌体的高稳定性及易检出性的天然优势,通过对其所携带miRNAs的表达量变化进行检测,可监测疾病进展和治疗效果,从而为结肠癌患者提供更为个性化的诊疗策略。
基金supported by the Basic Research Program of the Guizhou Science Cooperation Foundation Project(Grant Number:ZK[2021]466)Guizhou Provincial Health Commission(Grant Number:gzwkj2022-062).
文摘Background:Terpinen-4-ol(T4O),a key constituent of tea tree essential oil and various aromatic plants,has shown promising antiproliferative and pro-apoptotic effects in melanoma and other cancer types.However,its efficacy against cutaneous squamous cell carcinoma(cSCC)remains unclear.Thus,in this study,we investigated the in vivo and in vitro effects of T4O on cSCC cell lines and preliminarily explored its impacting pathways.Methods:Using CCK8 and assay colony formation,we assessed the viability of cSCC A431,SCL-1,and COLO-16 cells treated with T40 at varying concentrations(0,1,2,and 4μM).Flow cytometry was employed to evaluate T4O’s effect on cSCC cell’s cycle progression and apoptosis induction.Additionally,western blotting was utilized to examine the expression intensities of N-cadherin and E-cadherin,two indicative markers of the epithelial-mesenchymal transition(EMT)pathway.T4O’s in vivo effect on inhibiting tumor progression was evaluated on an established xenograft tumor model.Then,the molecular mechanisms of T4O’s antitumor effect were explored by an integrated genome-wide transcriptomics and proteomics study on cSCC A431c cells.Finally,calpain-2’s potential mediator role in T4O’s anti-tumor mechanism was investigated in calpain-2 knockdown cell lines prepared via siRNA transfection.Result:It’s demonstrated that T4O treatment inhibited cSCC proliferation,clonogenicity,migration,and invasion while inducing apoptosis and suppressing the EMT pathway.T4O administration also inhibited cSCC tumorigenesis in the xenograft tumor model.RNA-sequencing and iTRAQ analysis detected significant upregulation of calpain-2 expression in T4O-treated cSCC cells.Western blotting confirmed that T4O significantly increased calpain-2 expression and promoted proteolytic cleavage ofβ-catenin and caspase-12,two calpain-2 target proteins.Importantly,siRNA-mediated calpain-2 knockdown relieved T4O’s suppressive effect on cSCC cell proliferation and motility.Mechanistically,T4O upregulates calpain-2 expression and promotes the cleavage ofβ-catenin and caspase-12,with siRNA-mediated calpain-2 knockdown mitigating T4O’s suppressive effects.Conclusion:These findings suggest that T4O’s antitumor activity in cSCC is mediated through the upregulation of calpain-2 expression and subsequent modulation ofβ-catenin and caspase-12.
文摘End-stage kidney failure(ESKD)is a global issue where kidney replacement therapy imposes enormous economic burden to people of developing countries,in addition to the severe limitations to the availability of hemodialysis and peritoneal dialysis technique.The best option of kidney transplantation also requires lifelong combination immunosuppressive medicines,the cost of which is equally comparable to lifelong dialysis.A strategy of achieving transplant tolerance that requires minimum immunosuppressive medicines,although in experimental stage,also requires state-of-art technology with costly medicines and interventions.This is evidently beyond the reach of ESKD patients of developing countries.Hence,globally in developing countries,a need for an innovative but cost-effective tolerance protocol is a burning need for a successful transplant program.In brief,transplant tolerance is defined as a state of donorspecific unresponsiveness to the allograft antigens without the need for ongoing pharmacologic immunosuppression or with a minimal need.Current state-of-art techniques involves:(1)A state of hematological chimera,for complete tolerance;(2)Prope or partial tolerance where immune-reactive T-lymphocytes are inhibited using monoclonal antibodies;and(3)Chimeric antigen receptor for T-regulatory(T-reg)cell therapy using genetically engineered T-reg cells targeting specific Tlymphocyte receptors for inducing anergy.From our real-world experience in transplant management in post-transplant lympho-proliferative disorders(PTLD),we noticed frequently a drastic reduction in the need of immunosuppressive medicines following lympho-ablative therapy for PTLD.We recently published a case study on a real-world experience transplant case where we explained a partial or prope tolerance that developed after lymphocyte ablation therapy,following which the allograft was maintained with low dose dual standard immunosuppressive medicines.Based on this publication,we propose here an innovative tolerance protocol for living related low risk kidney transplantation for developing countries,in this opinion review.
文摘通过对100 t LF精炼37Mn5套管钢时1 540～1580℃钢水中稳定氧化物夹杂含量与T[O]的关系以及钢-渣平衡时渣碱度对钢中活性氧含量影响的分析,提出降低钢中T[O]和夹杂物含量的改进工艺措施。热力学计算和试验结果表明,当[Al]s为0.03%～0.04%,精炼渣碱度为3,(CaO)/(Al_2O_3)=3～3.5时,37Mn5钢中的氧含量降到10×10^(-6),管材基本消除了粗系夹杂,细系夹杂A+B+C+D≤5.0级,达到使用要求。
文摘T2O(Television to Online)是互联网+时代一种从电视到电商的新型营销传播模式。本研究结合创新扩散理论、SOR模型、计划行为理论、TAM模型等传播学、心理学、消费者行为学的跨学科理论,通过深度访谈、问卷调查等方式获取T2O营销传播模式下影响消费者行为意向的因素。发现在T2O模式下,消费者的感知态度、主观规范和个人创新性都会对其行为意向产生显著影响,其中感知娱乐的影响最大。本文为T2O的研究提供了新的角度,同时为电视台、电商和广告主深入实践应用T2O模式提出了建议。
