目的通过NHE1基因敲除模型鼠的海马组织差异蛋白质组学分析,发现并明确Ppp3cb和Ppm1g的表达特征。方法①选取6只2周龄NHE1基因敲除模型鼠作为模型组,同周龄野生型小鼠6只作为对照组,采用琼脂糖凝胶电泳检测其基因型;应用旷场实验和强迫...目的通过NHE1基因敲除模型鼠的海马组织差异蛋白质组学分析,发现并明确Ppp3cb和Ppm1g的表达特征。方法①选取6只2周龄NHE1基因敲除模型鼠作为模型组,同周龄野生型小鼠6只作为对照组,采用琼脂糖凝胶电泳检测其基因型;应用旷场实验和强迫游泳实验对模型组和对照组小鼠进行行为学评估,并按照Racine评分标准对模型鼠进行癫痫发作分级;②通过串联质谱分析技术对模型组和对照组的海马组织进行差异蛋白筛选,基因本体论(Gene Ontology Analysis,GO)分析差异蛋白并进行注释和富集,蛋白网络数据库(search tool for the retrieval of interesting genes,STRING)分析差异蛋白之间的蛋白相互作用(protein-protein interaction,PPI);③应用qPCR和Western blot检测Ppp3cb和Ppm1g的转录和翻译水平,应用免疫组织化学技术分别观察其在组织中的表达量。结果①模型组小鼠NHE1基因未见表达,旷场实验中模型鼠的运动总距离较对照组减少(P=0.0073),跨越的格子数比对照组显著减少(P<0.0001)。强迫游泳实验结果显示,模型鼠不动的时间明显延长(P<0.0001);②以表达倍数(FC)≥1.2倍且P<0.05为筛选标准,检测到海马组织中845个差异表达的蛋白质点,其中有9个蛋白表达上调,7个蛋白表达下调。其中Ppp3cb下调,Ppm1g上调。GO功能注释结果表明,NHE1敲除后,分子功能(MF)富集在蛋白丝氨酸/苏氨酸磷酸酶活性的差异最显著,细胞成分(CC)富集在质膜部分的差异蛋白数量最多,生物过程(BP)富集在负向调节生物过程、免疫系统过程的差异蛋白数量最多。STRING分析显示差异蛋白Ppp3cb和Slc9a1直接作用,Ppm1g通过Ppp3cb和Slc9a1间接作用,Ppp3cb和Ppm1g之间相互作用。③Ppp3cb的转录和翻译水平减少,在组织中的表达量下降,而Ppm1g转录和翻译水平增加,在组织中的表达量上升(P<0.05)。结论本研究确定了NHE1基因敲除小鼠海马组织中差异蛋白Ppp3cb表达下调,而Ppm1g表达上调,为进一步研究Ppp3cb和Ppm1g参与癫痫发病机制提供了依据。展开更多
目的:检测PIK3CA、PIK3CB和多药耐药基因1(muhidrug resistance 1,MDR-1)、肺耐药蛋白(lung cancer resistance protein,LRP)、谷胱甘肽转移酶π(glutathione-S-transferaseπ,GST-π)、D N A拓扑异构酶Ⅱ(DNA topoisomeraseⅡ,TopoⅡ)...目的:检测PIK3CA、PIK3CB和多药耐药基因1(muhidrug resistance 1,MDR-1)、肺耐药蛋白(lung cancer resistance protein,LRP)、谷胱甘肽转移酶π(glutathione-S-transferaseπ,GST-π)、D N A拓扑异构酶Ⅱ(DNA topoisomeraseⅡ,TopoⅡ)在大肠癌中的表达,探讨PIK3CA、PIK3CB对大肠癌多药耐药性的影响.方法:应用免疫组织化学EnVision法检测316例大肠癌组织中PIK3CA、PIK3CB、MDR-1、LRP、GST-π和TopoⅡ的表达情况,并结合临床病理因素进行分析.结果:316例大肠癌中,PIK3CA、PIK3CB的表达与肿瘤的分化程度、淋巴结转移成正相关,其相关系数分别为0.136、0.168.MDR基因在大肠癌组织中的阳性率分别为:M D R-1为72.78%(230/316)、L R P为70.89%(224/316)、TOPⅡ为77.53%(245/316)及GST-π为76.58%(242/316).经Spearman相关性分析,PIK3CA、PIK3CB的表达与多药耐药蛋白MDR-1、LRP、GST-π的表达均成正相关,相关系数分别为0.288、0.128、0.1 9 7.3 1 6例大肠癌患者的中位生存期为60 mo,5年生存率为47.5%,Kaplan-Meier分析结果显示,PIK3CA与PIK3CB同时高表达、淋巴结转移、MDR-1(+)、GST-π(+)患者术后5年生存率明显降低.Cox比例风险模型显示,PIK3CA(+)PIK3CB(+)、淋巴结转移、MDR-1、GST-π是影响大肠癌患者预后的独立因素.结论:PIK3CA、PIK3CB与大肠癌的多药耐药密切相关,PIK3CA、PIK3CB、淋巴结转移、MDR-1、GST-π是影响大肠癌患者预后的独立因素,检测PIK3CA、PIK3CB为临床合理选择化疗药物、初步判断预后具有重要参考价值.展开更多
Class I phosphoinositide 3-kinase(PI3K)enzymes have attracted considerable attention as drug targets in cancer therapy over the last 20 years.The signaling pathway triggered by class I PI3Ks is dysregulated in a range...Class I phosphoinositide 3-kinase(PI3K)enzymes have attracted considerable attention as drug targets in cancer therapy over the last 20 years.The signaling pathway triggered by class I PI3Ks is dysregulated in a range of tumor types,impacting cell proliferation,survival and apoptosis.Frequent oncogenic mutations of PIK3CA have previously been discovered.In contrast,reports of PIK3CB mutations have been limited;however,in most cases,those that have been identified have been shown to be activating and oncogenic.The functional characterization of a PIK3CB catalytic domain mutant,p110β^(E1051K),first discovered by others in castrateresistant prostate cancer(mCRPC),is outlined in this report;our data suggest that p110β^(E1051K)is a gain-of-function mutation,driving PI3K signaling,tumorigenic cell growth and migration.Tumor cells expressing p110β^(E1051K)are sensitive to p110βinhibition;its characterization as an oncogenic driver adds to the rationale for targeting p110βand indicates a continuing need to further develop specific PI3K inhibitors for clinical development in cancer therapy.展开更多
A new monomer,vinyltriallylsilane（CB3）,having functionnal groups with different hydrosilylation reactivity was designed and synthesized by allylation reaction from vinyltrichlorosilane.Then,photocrosslinkable hyperb...A new monomer,vinyltriallylsilane（CB3）,having functionnal groups with different hydrosilylation reactivity was designed and synthesized by allylation reaction from vinyltrichlorosilane.Then,photocrosslinkable hyperbranched（poly（siloxysilane）） with terminal allyl groups was prepared via hydrosilylation reaction of vinyltriallylsilane（CB3） and dimethylbis（dimethylsiloxy）silane（A2） monomers at the presence of Karstedt catalyst,and characterized by means of FTIR,（）1H-NMR,（）29Si-NMR,and SEC/RI/MALLS technology.The polymerization process of A2 and CB3 monomers was studied in situ by using FTIR.It was found that silicon hydride would preferentially react with vinyl groups during the reaction. This can cause to the generation of an intermediate with one Si—H and three allyl groups.By using it to conduct further self-polymerization,hyperbranched polymer can be obtained.The degree of branching of the resulting hyperbranched poly（siloxysilane） was calculated to be 0.44 by quantitative（）29Si-NMR spectroscopy,and the weight average molecular weight was 12.1 kg/mol with its polydispersity of 2.52.Finally,the UV curing behavior of the resulting polymer initiated with different photoinitors was also investigated.展开更多
文摘目的通过NHE1基因敲除模型鼠的海马组织差异蛋白质组学分析,发现并明确Ppp3cb和Ppm1g的表达特征。方法①选取6只2周龄NHE1基因敲除模型鼠作为模型组,同周龄野生型小鼠6只作为对照组,采用琼脂糖凝胶电泳检测其基因型;应用旷场实验和强迫游泳实验对模型组和对照组小鼠进行行为学评估,并按照Racine评分标准对模型鼠进行癫痫发作分级;②通过串联质谱分析技术对模型组和对照组的海马组织进行差异蛋白筛选,基因本体论(Gene Ontology Analysis,GO)分析差异蛋白并进行注释和富集,蛋白网络数据库(search tool for the retrieval of interesting genes,STRING)分析差异蛋白之间的蛋白相互作用(protein-protein interaction,PPI);③应用qPCR和Western blot检测Ppp3cb和Ppm1g的转录和翻译水平,应用免疫组织化学技术分别观察其在组织中的表达量。结果①模型组小鼠NHE1基因未见表达,旷场实验中模型鼠的运动总距离较对照组减少(P=0.0073),跨越的格子数比对照组显著减少(P<0.0001)。强迫游泳实验结果显示,模型鼠不动的时间明显延长(P<0.0001);②以表达倍数(FC)≥1.2倍且P<0.05为筛选标准,检测到海马组织中845个差异表达的蛋白质点,其中有9个蛋白表达上调,7个蛋白表达下调。其中Ppp3cb下调,Ppm1g上调。GO功能注释结果表明,NHE1敲除后,分子功能(MF)富集在蛋白丝氨酸/苏氨酸磷酸酶活性的差异最显著,细胞成分(CC)富集在质膜部分的差异蛋白数量最多,生物过程(BP)富集在负向调节生物过程、免疫系统过程的差异蛋白数量最多。STRING分析显示差异蛋白Ppp3cb和Slc9a1直接作用,Ppm1g通过Ppp3cb和Slc9a1间接作用,Ppp3cb和Ppm1g之间相互作用。③Ppp3cb的转录和翻译水平减少,在组织中的表达量下降,而Ppm1g转录和翻译水平增加,在组织中的表达量上升(P<0.05)。结论本研究确定了NHE1基因敲除小鼠海马组织中差异蛋白Ppp3cb表达下调,而Ppm1g表达上调,为进一步研究Ppp3cb和Ppm1g参与癫痫发病机制提供了依据。
文摘目的:检测PIK3CA、PIK3CB和多药耐药基因1(muhidrug resistance 1,MDR-1)、肺耐药蛋白(lung cancer resistance protein,LRP)、谷胱甘肽转移酶π(glutathione-S-transferaseπ,GST-π)、D N A拓扑异构酶Ⅱ(DNA topoisomeraseⅡ,TopoⅡ)在大肠癌中的表达,探讨PIK3CA、PIK3CB对大肠癌多药耐药性的影响.方法:应用免疫组织化学EnVision法检测316例大肠癌组织中PIK3CA、PIK3CB、MDR-1、LRP、GST-π和TopoⅡ的表达情况,并结合临床病理因素进行分析.结果:316例大肠癌中,PIK3CA、PIK3CB的表达与肿瘤的分化程度、淋巴结转移成正相关,其相关系数分别为0.136、0.168.MDR基因在大肠癌组织中的阳性率分别为:M D R-1为72.78%(230/316)、L R P为70.89%(224/316)、TOPⅡ为77.53%(245/316)及GST-π为76.58%(242/316).经Spearman相关性分析,PIK3CA、PIK3CB的表达与多药耐药蛋白MDR-1、LRP、GST-π的表达均成正相关,相关系数分别为0.288、0.128、0.1 9 7.3 1 6例大肠癌患者的中位生存期为60 mo,5年生存率为47.5%,Kaplan-Meier分析结果显示,PIK3CA与PIK3CB同时高表达、淋巴结转移、MDR-1(+)、GST-π(+)患者术后5年生存率明显降低.Cox比例风险模型显示,PIK3CA(+)PIK3CB(+)、淋巴结转移、MDR-1、GST-π是影响大肠癌患者预后的独立因素.结论:PIK3CA、PIK3CB与大肠癌的多药耐药密切相关,PIK3CA、PIK3CB、淋巴结转移、MDR-1、GST-π是影响大肠癌患者预后的独立因素,检测PIK3CA、PIK3CB为临床合理选择化疗药物、初步判断预后具有重要参考价值.
文摘Class I phosphoinositide 3-kinase(PI3K)enzymes have attracted considerable attention as drug targets in cancer therapy over the last 20 years.The signaling pathway triggered by class I PI3Ks is dysregulated in a range of tumor types,impacting cell proliferation,survival and apoptosis.Frequent oncogenic mutations of PIK3CA have previously been discovered.In contrast,reports of PIK3CB mutations have been limited;however,in most cases,those that have been identified have been shown to be activating and oncogenic.The functional characterization of a PIK3CB catalytic domain mutant,p110β^(E1051K),first discovered by others in castrateresistant prostate cancer(mCRPC),is outlined in this report;our data suggest that p110β^(E1051K)is a gain-of-function mutation,driving PI3K signaling,tumorigenic cell growth and migration.Tumor cells expressing p110β^(E1051K)are sensitive to p110βinhibition;its characterization as an oncogenic driver adds to the rationale for targeting p110βand indicates a continuing need to further develop specific PI3K inhibitors for clinical development in cancer therapy.
文摘A new monomer,vinyltriallylsilane（CB3）,having functionnal groups with different hydrosilylation reactivity was designed and synthesized by allylation reaction from vinyltrichlorosilane.Then,photocrosslinkable hyperbranched（poly（siloxysilane）） with terminal allyl groups was prepared via hydrosilylation reaction of vinyltriallylsilane（CB3） and dimethylbis（dimethylsiloxy）silane（A2） monomers at the presence of Karstedt catalyst,and characterized by means of FTIR,（）1H-NMR,（）29Si-NMR,and SEC/RI/MALLS technology.The polymerization process of A2 and CB3 monomers was studied in situ by using FTIR.It was found that silicon hydride would preferentially react with vinyl groups during the reaction. This can cause to the generation of an intermediate with one Si—H and three allyl groups.By using it to conduct further self-polymerization,hyperbranched polymer can be obtained.The degree of branching of the resulting hyperbranched poly（siloxysilane） was calculated to be 0.44 by quantitative（）29Si-NMR spectroscopy,and the weight average molecular weight was 12.1 kg/mol with its polydispersity of 2.52.Finally,the UV curing behavior of the resulting polymer initiated with different photoinitors was also investigated.
