目的评估胃泌素释放肽前体(Pro-gastrin-releasing peptide,PRO-GRP)、神经元特异性烯醇酶(Neuron-specific enolase,NSE)、癌胚抗原(Carcinoembryonic antigen,CEA)、细胞角蛋白-19片段21-1(Cytokeratin-19 fragment 21-1,CYFRA21-1)...目的评估胃泌素释放肽前体(Pro-gastrin-releasing peptide,PRO-GRP)、神经元特异性烯醇酶(Neuron-specific enolase,NSE)、癌胚抗原(Carcinoembryonic antigen,CEA)、细胞角蛋白-19片段21-1(Cytokeratin-19 fragment 21-1,CYFRA21-1)及鳞状细胞癌抗原(Squamous cell carcinoma antigen,SCC)血清肿瘤标志物在肺癌早期诊断中的价值,并探究它们作为诊断指标的组合效应。方法选择2020年1月至2022年10月期间收治的60例肺癌患者作为阳性组,同时纳入90例良性肺疾病患者作为阴性组。所有患者均接受血清肿瘤标志物检测,检测方法为免疫化学发光法,检测指标包括:PRO-GRP、NSE、CEA、CYFRA21-1及SCC。比较两组患者血清标志物的含量,并分析其阴阳表达情况。利用二元多因素回归分析,将这些标志物作为自变量,是否诊断为肺癌阳性作为因变量,探究其对肺癌诊断的独立危险因素。应用ROC曲线分析评估单独及联合检测这些标志物对肺癌的诊断价值,统计ROC曲线下面积(Area Under the Curve,AUC)、敏感度和特异度。结果阳性组患者的血清PRO-GRP、NSE、CEA、CYFRA21-1、SCC含量均显著高于阴性组(P<0.05)。单因素分析结果显示,这些标志物均为影响肺癌诊断结果的可疑因素(P<0.05)。多因素分析结果表明,它们在肺癌诊断为阳性时均为独立的危险因素(P<0.05)。ROC曲线分析显示,PRO-GRP、NSE、CEA、CYFRA21-1、SCC的单独及联合检测在肺癌诊断中具有统计学意义(P<0.05)。联合预测因子的AUC、敏感度和特异度均高于单独检测指标。结论PRO-GRP、NSE、CEA、CYFRA21-1及SCC血清肿瘤标志物在肺癌早期诊断中具有重要的诊断价值。联合检测这可以提高肺癌早期诊断的准确性,成为有力的肺癌筛查和早期诊断工具。展开更多
Objective:To investigate the regulatory effect of Qiliqiangxin Capsule on mitochondrial Ca^(2+)related genes in rats with myocardial infarction(MI).Methods:The rat model of MI was established by ligation of the left a...Objective:To investigate the regulatory effect of Qiliqiangxin Capsule on mitochondrial Ca^(2+)related genes in rats with myocardial infarction(MI).Methods:The rat model of MI was established by ligation of the left anterior descending coronary artery.After operation,the rats were randomly assigned to the model group,the Qiliqiangxin group and the captopril group;a sham-operated group was also available as a control.After four weeks of treatment,the extent of infarction in rats was observed by gross cardiac structure and the morphological changes of myocardial histopathology were observed by HE staining.Detection of mitochondrial Ca^(2+)transport-related genes such as inositol-1,4,5-trisphosphate receptor 2(IP3R2),glucose regulated protein 75(GRP75),voltage-dependent anion channel 1(VDAC1),and mitofusion 2(Mfn2)and mitochondrial apoptosis-related genes such as B-cell lymphoma-2(Bcl-2)and Bcl-2 related X protein(Bax)mRNA expression changes was measured by RT-PCR in the infarct margins of the heart;Western blot was used to detect changes in Bcl-2,Bax protein expression in myocardial tissue.The rate of apoptosis in cardiac myocardial tissue was detected by TUNEL staining.Results:Compared with the sham group,the anterior left ventricular wall of the model group showed a large area of infarction,and the structure of myocardial tissue was disordered.The mRNA expression level of mitochondrial Ca^(2+)transport-related genes such as IP3R2,GRP75,VDAC1,and Mfn2 were significantly increased(P<0.05,P<0.01);The mRNA and protein expression of Bcl-2,a molecule related to mitochondrial apoptosis,were significantly decreased(P<0.01),while the mRNA and protein expression of Bax were significantly increased(P<0.01);and apoptosis rate was significantly increased(P<0.01).Compared with the model group,the infarct size of cardiac gross specimens in the Qiliqiangxin group and the captopril group was reduced and myocardial fibers were relatively well ordered;The mRNA expression of mitochondrial Ca^(2+)transport-related genes such as IP3R2,GRP75,VDAC1,and Mfn2 were significantly reduced(P<0.01);the mRNA and protein expression of Bcl-2,a molecule related to mitochondrial apoptosis,were increased(P<0.05,P<0.01),and the mRNA and protein expression of Bax were significantly decreased(P<0.05,P<0.01).and apoptosis rate was significantly decreased(P<0.01).Conclusion:Qiliqiangxin Capsule can improve the morphological structure of the heart of rats with MI,and its mechanism is related to regulation of the gene expression of mitochondrial Ca^(2+)transport complex IP3R2/GRP75/VDAC1,thereby inhibiting apoptosis.展开更多
文摘【背景】随着规模化、集约化生产程度的不断提高,养殖过程中饲养空间受限、冷热环境不适等因素常使猪处于应激状态。内质网应激(endoplasmic reticulum stress,ERS)可能是最早期的应激反应,与细胞凋亡、代谢等方面有密切联系。肝脏是机体的主要代谢器官,猪养殖过程中由于人工操作(如断奶)、饲料霉变、温度变化和吸入有害气体等因素都会造成猪肝脏的ERS,不仅会造成肝脏损伤,还会引发肝脏的脂肪代谢紊乱和广泛的炎症反应,影响生产性能和繁殖性能。因此,深入探讨缓解ERS的有效措施,有助于减少猪养殖过程中的隐性损失。【目的】利用免疫沉淀联合质谱技术,从猪肝星状细胞中筛选在ERS条件下与葡萄糖调节蛋白94(GRP94)相互作用的细胞蛋白,为进一步探讨GRP94对猪肝星状细胞生物学功能的保护作用机理奠定基础。【方法】首先将GRP94抗体固定在谷胱甘肽亲和磁珠上,用亲和磁珠与ERS条件下或正常条件下猪肝星状细胞总蛋白进行孵育,与GRP94诱饵蛋白结合的蛋白复合物洗脱收集后,进行SDS-PAGE凝胶电泳验证。将验证成功的样品洗脱液进行液相色谱串联质谱(LC-MS/MS)检测,鉴定出正常条件和ERS条件下GRP94的互作蛋白。运用生物信息学在线软件对筛选的互作细胞蛋白进行GO富集、KEGG信号通路注释和蛋白互作网络分析,并对其中的互作蛋白之一波形蛋白(vimentin)进行免疫共沉淀验证。【结果】筛选到正常条件下与GRP94存在互作关系的蛋白146个,ERS条件下与GRP94存在互作关系的蛋白76个,在两种情况下都存在互作关系的蛋白44个。ERS条件下有互作关系的76个蛋白质主要参与凋亡过程负调控、肽段交联、泛素依赖型ERAD(endoplasmic reticulum associated degradation)过程和过氧化氢分解代谢等过程。其中参与凋亡过程负调控的GRP94互作蛋白有albumin、catalase、filament A、heat shock protein family A member 5、keratin 18和prohibin 2,说明GRP94可能与这些蛋白共同发挥抗凋亡作用。除此之外组成中间丝纤维的vimentin蛋白参与多个GO富集的通路,可能与GRP94有重要的互作关系。进一步的免疫共沉淀试验也证实,ERS条件下vimentin和GRP94之间确实存在互作关系。此外,某些ERS条件下特异性表达的GRP94互作蛋白(如peroxiredoxin、death inducer obliterator 1、catalase、glandular kallikrein、pyruvate kinase等)与抗凋亡有密切联系。【结论】ERS条件下,猪肝脏GRP94互作蛋白主要参与抗凋亡、对未折叠蛋白进行折叠和维护细胞内稳态相关的信号通路。该结论为下一步开展GRP94参与肝脏ERS调控机制的研究打下基础。
文摘目的评估胃泌素释放肽前体(Pro-gastrin-releasing peptide,PRO-GRP)、神经元特异性烯醇酶(Neuron-specific enolase,NSE)、癌胚抗原(Carcinoembryonic antigen,CEA)、细胞角蛋白-19片段21-1(Cytokeratin-19 fragment 21-1,CYFRA21-1)及鳞状细胞癌抗原(Squamous cell carcinoma antigen,SCC)血清肿瘤标志物在肺癌早期诊断中的价值,并探究它们作为诊断指标的组合效应。方法选择2020年1月至2022年10月期间收治的60例肺癌患者作为阳性组,同时纳入90例良性肺疾病患者作为阴性组。所有患者均接受血清肿瘤标志物检测,检测方法为免疫化学发光法,检测指标包括:PRO-GRP、NSE、CEA、CYFRA21-1及SCC。比较两组患者血清标志物的含量,并分析其阴阳表达情况。利用二元多因素回归分析,将这些标志物作为自变量,是否诊断为肺癌阳性作为因变量,探究其对肺癌诊断的独立危险因素。应用ROC曲线分析评估单独及联合检测这些标志物对肺癌的诊断价值,统计ROC曲线下面积(Area Under the Curve,AUC)、敏感度和特异度。结果阳性组患者的血清PRO-GRP、NSE、CEA、CYFRA21-1、SCC含量均显著高于阴性组(P<0.05)。单因素分析结果显示,这些标志物均为影响肺癌诊断结果的可疑因素(P<0.05)。多因素分析结果表明,它们在肺癌诊断为阳性时均为独立的危险因素(P<0.05)。ROC曲线分析显示,PRO-GRP、NSE、CEA、CYFRA21-1、SCC的单独及联合检测在肺癌诊断中具有统计学意义(P<0.05)。联合预测因子的AUC、敏感度和特异度均高于单独检测指标。结论PRO-GRP、NSE、CEA、CYFRA21-1及SCC血清肿瘤标志物在肺癌早期诊断中具有重要的诊断价值。联合检测这可以提高肺癌早期诊断的准确性,成为有力的肺癌筛查和早期诊断工具。
基金This study was supported by Beijing University of Traditional Chinese Medicine Dongzhimen Hospital 2022 Science and Technology Innovation Special Project(DZMKJCX-2022-008)。
文摘Objective:To investigate the regulatory effect of Qiliqiangxin Capsule on mitochondrial Ca^(2+)related genes in rats with myocardial infarction(MI).Methods:The rat model of MI was established by ligation of the left anterior descending coronary artery.After operation,the rats were randomly assigned to the model group,the Qiliqiangxin group and the captopril group;a sham-operated group was also available as a control.After four weeks of treatment,the extent of infarction in rats was observed by gross cardiac structure and the morphological changes of myocardial histopathology were observed by HE staining.Detection of mitochondrial Ca^(2+)transport-related genes such as inositol-1,4,5-trisphosphate receptor 2(IP3R2),glucose regulated protein 75(GRP75),voltage-dependent anion channel 1(VDAC1),and mitofusion 2(Mfn2)and mitochondrial apoptosis-related genes such as B-cell lymphoma-2(Bcl-2)and Bcl-2 related X protein(Bax)mRNA expression changes was measured by RT-PCR in the infarct margins of the heart;Western blot was used to detect changes in Bcl-2,Bax protein expression in myocardial tissue.The rate of apoptosis in cardiac myocardial tissue was detected by TUNEL staining.Results:Compared with the sham group,the anterior left ventricular wall of the model group showed a large area of infarction,and the structure of myocardial tissue was disordered.The mRNA expression level of mitochondrial Ca^(2+)transport-related genes such as IP3R2,GRP75,VDAC1,and Mfn2 were significantly increased(P<0.05,P<0.01);The mRNA and protein expression of Bcl-2,a molecule related to mitochondrial apoptosis,were significantly decreased(P<0.01),while the mRNA and protein expression of Bax were significantly increased(P<0.01);and apoptosis rate was significantly increased(P<0.01).Compared with the model group,the infarct size of cardiac gross specimens in the Qiliqiangxin group and the captopril group was reduced and myocardial fibers were relatively well ordered;The mRNA expression of mitochondrial Ca^(2+)transport-related genes such as IP3R2,GRP75,VDAC1,and Mfn2 were significantly reduced(P<0.01);the mRNA and protein expression of Bcl-2,a molecule related to mitochondrial apoptosis,were increased(P<0.05,P<0.01),and the mRNA and protein expression of Bax were significantly decreased(P<0.05,P<0.01).and apoptosis rate was significantly decreased(P<0.01).Conclusion:Qiliqiangxin Capsule can improve the morphological structure of the heart of rats with MI,and its mechanism is related to regulation of the gene expression of mitochondrial Ca^(2+)transport complex IP3R2/GRP75/VDAC1,thereby inhibiting apoptosis.
