检索结果-南通市图书馆

Extraction, detection, and profiling of serum biomarkers using designed Fe3O4@SiO2@HA core-shell particles

在线全文

学校读者我要写书评

暂无评论

Nano Research 2018年第1期11卷 68-79页

作者： Chandrababu Rejeeth Xuechao Pang Ru Zhang Wei Xu Xuming Sun Bin Liu jiatao lou Jingjing Wan Hongchen Gu Wei Yan Kun Qian School of Biomedical Engineering Shanghai Center for System Biomedicine Shanghai Chest Hospital and Med-X Research Institute Shanghai Jiao Tong University Shanghai 200030 China Department of Chemistry Shanghai University Shanghai200444 China

Serum biomarkers in the form of proteins （e.g. cluster of differentiation-44 （CD44）） have been demonstrated to have high clinical sensitivity and specificity for disease diagnosis and prognosis. Owing to the high sample complexity and low molecular abundance in serum, the detection and profiling of biomarkers rely on efficient extraction by materials and devices, mostly using immunoassays via antibody-antigen recognition. Antibody-free approaches are promising and need to be developed for real-case applications in serum to address the limitations of antibody-based techniques in terms of robustness, expense, and throughput. In this work, we demonstrated a novel approach using hyaluronic acid （HA）-modified materials/devices for the extraction, detection, and profiling of serum biomarkers via ligand-protein interactions. We constructed Fe304@SiOa@HA particles with different sizes through layer-by-layer assembly and for the first time applied HA-functionalized particles in the facile extraction and sequence identification of CD44 in serum by mass spectrometry. We also first validated HA-CD44 binding through electrochemical sensing using HA- modified electrodes in both standard solutions and diluted serum samples, achieving a detection limit of -0.6 ng/mL and a linear response range from I ng/mL to 10 ~tg/mL. Furthermore, we performed profiling of HA-binding serum proteome, providing a new preliminary benchmark for the construction of future databases, and we investigated selected surface chemistries of particles for the capture of proteins in serum. Our work not only resulted in the development of a platform technology for CD44 extraction/detection and HA-binding proteome identification, but also guided the design of ligand affinity-based approaches for antibody-free analysis of serum biomarkers towards diagnostic applications.

关键词： ligand-protein interaction magnetic particles serum biomarkers cluster of differentiation44(CD44) sensors mass spectrometry

Methylation analysis of SHOX2 and RASSF1A panel in bronchoalveolar lavage fluid for lung cancer diagnosis

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Methylation analysis of SHOX2 and RASSF1A panel in bronchoal...

Evaluation of the performance of EGFR mutations detection in advanced NSCLC patientswith droplet digital PCR

2017年中国肿瘤标志物学术大会暨第十一届肿瘤标志物青年科学家论坛

作者： Chenzi Zhang Wenjun Yu jiatao lou Shanghai chest hospital

Introduction: Since currently no sensitive and specific biomarkers for lung cancer diagnosis and the majority of lung cancer patients are diagnosed at late stage of diseases, noninvasive biomarkers with high sensitivity and specificity are urgently needed in lung cancer diagnosis. Thus, we examined the aberrant methylation of the SHOX2 and RASSF1 A panel in comparing with conventional cytology examination and CEA in order to identify the diagnosing value of the novel panel. Patients and Methods: Bronchoalveolar lavage fluid(BALF) and serum samples were collected from 322 patients, 284 of them finally diagnosed lung cancer, and 38 were control patients including benign lung diseases and malignancies not in lung. For all of the 322 patients, real-time methylation specific PCR assay and sanger sequencing were performed using BALF to detect the aberrant methylation of SHOX2 and RASSF1 A, serum CEA were detected using electrochemiluminescence immunoassay kit. Cytology and pathology examinations were performed by Pathology ***: Profiling data showed the consistency of RT-PCR and sanger sequencing in detecting of the methylation of SHOX2 and RASSF1 A panel was satisfactory. The SHOX2+RASSF1 A panel revealed a satisfying diagnosis value(sensitivity=81.0%, specificity=97.4%) in lung cancer diagnosis in comparing with CEA(sensitivity=31.0%, specificity=100.0%) and cytology examination(sensitivity=68.3%, specificity=97.4%). The positive detection rate of the SHOX2+RASSF1 A panel was 90.5% in small lung cancer, higher than in squamous cell carcinoma(88.8%) and in adenocarcinoma(69.6%).Conclusion: The methylation analysis of SHOX2+RASSF1 A panel could be used as potential noninvasive biomarkers for lung cancer diagnosis.

关键词：

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

Evaluation of the performance of EGFR mutations detection in...

2017年中国肿瘤标志物学术大会暨第十一届肿瘤标志物青年科学家论坛

作者： QiaomeiGuo Lin Wang Wenjun Yu PeijunZhai jiatao lou Shanghai chest hospital

Objective:To investigate the feasibility of droplet digital PCR(ddPCR) for the detection of 3 EGFR mutations(exon 19 deletions, L858 R in exon 21, and T790 M in exon 20)from plasma ctDNAin advancednon-small cell lung cancer( NSCLC) patients. The accuracyofddPCR wasevaluated bycomparing with 2 different ***: The sensitivity, specificity,accuracy of the established ddPCR system were assessed using purified EGFR mutation-positive cell DNA and reference standard genome *** performance of our ddPCR systemfor detecting EGFR mutations in plasma ctDNA was compared by paralleltestingwith other 2 different platforms(ARMS and NGS). Finally,ourddPCR system was used to detect the plasma EFGR mutations in targeted treatment-na?ve NSCLC patients and those who previously undergoneEGFR-TKI therapy. Results: Our establishedddPCR system showed 100% specifityandhigh concordance for EGFR mutation detection. The assay sensitivity of ddPCR was 0.1% for EGFR 19 deletions and L858 R,whereas 0.5% for T790 *** testing 64 plasma cfDNA,bothNGS and ddPCR displayed superior sensitivity than ARMS(P<0.01).Of 159 patients advanced NSCLC with molecular pathology results, 78 cases were EGFR-TKI treatment-naive and the other 81 wereundergone bloodrebiopsy after acquired *** ddPCR also exhibit a good concordance inclinical ***: We therefore developed ahighly sensitive and specific droplet digital PCR method for plasma EGFR mutation(exon19 deletions, L858 R andT790 M) testing.

关键词： droplet digital PCR EGFR non-small cell lung cancer ctDNA

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

循环肿瘤DNA的研究进展

中国肿瘤生物治疗杂志 2016年第5期23卷 601-608页

作者：郭巧梅娄加陶上海交通大学附属上海胸科医院检验科上海200030

循环肿瘤DNA(circulating tumor DNA,ctDNA)是由肿瘤细胞释放到循环系统中的基因组小片段,其来源于机体所有荷瘤部位,具有含量极低、个体间差异大、半衰期短、匀质性、携带有肿瘤特异性遗传学/表观遗传学变异等特点,与肿瘤的发展进化、... 详细信息

循环肿瘤DNA(circulating tumor DNA,ctDNA)是由肿瘤细胞释放到循环系统中的基因组小片段,其来源于机体所有荷瘤部位,具有含量极低、个体间差异大、半衰期短、匀质性、携带有肿瘤特异性遗传学/表观遗传学变异等特点,与肿瘤的发展进化、休眠耐药、转移复发等密切相关,正逐渐成为肿瘤学分子研究领域的热点。目前针对ctDNA的检测方法和平台种类繁多,主要分为基于PCR和二代测序(next-generation sequence,NGS)的方法,二者各有利弊,需要根据研究目的灵活选择并建立统一标准。随着相关技术的发展和法规的不断完善,ctDNA可作为癌症筛查、早期诊断、个体化治疗及预后评估理想的血源性生物标志物,在肿瘤的精准医疗中必将大放异彩。

关键词：循环肿瘤DNA 肿瘤表观遗传学变异二代测序

Comparative analysis of QS3D and droplet digital PCR for detection of plasma EGFR T790M mutation at disease progression

在线全文

学校读者我要写书评

暂无评论

Comparative analysis of QS3D and droplet digital PCR for det...

Integrated microfluidic single-cell immunoblotting chip enables high-throughput isolation,enrichment and direct protein analysis of circulating tumor cells

2021年中国肿瘤标志物学术大会暨第十五届肿瘤标志物青年科学家论坛

作者： Qiaomei Guo Xia Ahung Mingna Zhao Mengyan Wang jiatao lou Department of Laboratory Medicine Shanghai Chest Hospital

Objectives: Numerous studies have demonstrated the ability of QuantStudioTM 3 D(QS3 D) and droplet digital PCR(dPCR) platforms in the successful detection of EGFR mutations from plasma. However,no prospective data on quantitative comparative analysis of the two methods are available. Here we directly compared the performance of the two platforms in the detection of the EGFR T790 M mutation in cell-free DNA(cfDNA) samples from lung cancer ***: cfDNA was isolated from plasma samples of 72 non-small cell lung cancer patients who initially received tyrosine kinase inhibitor treatment and acquired drug resistance. Identical cfDNA samples were subjected to T790 M mutation analysis using QS3 D and droplet dPCR in ***: The T790 M mutation was detected in 15/72(20.8%) cfDNA samples by QS3 D dPCR and 21/72(29.2%) cfDNA samples by droplet dPCR. The overall concordance between the two methods was91.7%(66/72). The median allele frequency of the T790 M mutation detected by QS3 D dPCR and droplet dPCR was 2.01% and 2.62%, respectively. The partition number of QS3 D dPCR is much higher than that of droplet dPCR, while the mutation abundance detected by the two platforms is similar. Furthermore, a strong correlation was observed in allele frequencies and copy numbers between the two methods, with R2 of 0.98 and 0.97, ***: Our study demonstrated a high level of concordance between the QS3 D and droplet dPCR platforms for the detection and absolute quantification of the EGFR T790 M mutation in cfDNA.

关键词： cfDNA T790M digital PCR concordance mutation abundance

来源： cnki会议评论

在线全文

cnki会议

学校读者我要写书评

暂无评论

Microsystems & Nanoengineering 2022年第1期8卷 211-222页

作者： Aynur Abdulla Ting Zhang Shanhe Li Wenke Guo Antony RWarden Yufang Xin Nokuzola Maboyi jiatao lou Haiyang Xie Xianting Ding Institute for Personalized Medicine School of Biomedical EngineeringShanghai Jiao Tong UniversityShanghai 200030China Shanghai General Hospital Shanghai Jiao Tong UniversityNo.85 Wujing RoadShanghai 200080China

Effective capture and analysis of a single circulating tumor cell(CTC)is instrumental for early diagnosis and personalized therapy of ***,due to their extremely low abundance and susceptibility to interference from other cells,high-throughput isolation,enrichment,and single-cell-level functional protein analysis of CTCs within one integrated system remains a major ***,we present an integrated multifunctional microfluidic system for highly efficient and label-free CTC isolation,CTC enrichment,and single-cell immunoblotting(ieSCI).The ieSCI-chip is a multilayer microfluidic system that combines an inertia force-based cell sorter with a membrane filter for label-free CTC separation and enrichment and a thin layer of a photoactive polyacrylamide gel with microwell arrays at the bottom of the chamber for single-cell *** ieSCI-chip successfully identified a subgroup of apoptosis-negative(Bax-negative)cells,which traditional bulk analysis did not detect,from cisplatin-treated ***,we demonstrated the clinical application of the ieSCI-chip with blood samples from breast cancer patients for personalized CTC epithelial-to-mesenchymal transition(EMT)*** expression level of a tumor cell marker(EpCAM)can be directly determined in isolated CTCs at the single-cell level,and the therapeutic response to anticancer drugs can be simultaneously ***,the ieSCI-chip provides a promising clinical translational tool for clinical drug response monitoring and personalized regimen development.

关键词： diagnosis analysis enable

IQGAP1通过ERK信号通路促进非小细胞肺癌细胞增殖

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

中国细胞生物学学报 2016年第6期38卷 662-668页

作者：赵明娜娄加陶上海交通大学附属胸科医院检验科上海200030

该研究探讨了IQGAP1(IQ domain GTPase-activating protein 1)对非小细胞肺癌(nonsmall cell lung cancer,NSCLC)细胞增殖的影响及其对ERK信号通路的调节作用。将内源性IQGAP1低表达的A549细胞中分为空白组、空载组和IQGAP1过表达组;将内源性IQGAP1高表达的H1299细胞中分为空白组、阴性对照si RNA组和IQGAP1 si RNA组;采用ERK1/2磷酸化抑制剂U0126处理上述两株细胞。MTT法检测细胞增殖能力,Western blot法检测ERK1/2和p-ERK1/2蛋白质水平。结果显示,在A549细胞中,过表达IQGAP1能促进细胞增殖并促进ERK1/2磷酸化;在H1299中,敲低IQGAP1表达能够抑制细胞增殖并下调ERK1/2磷酸化水平。用U0126处理后能抑制IQGAP1对细胞增殖的促进作用。研究结果表明,IQGAP1可通过ERK信号通路促进体外非小细胞肺癌细胞增殖。

关键词：非小细胞肺癌 IQGAP1 ERK 信号通路细胞增殖

miR-93通过靶向EphA4激活ERK通路促进非小细胞肺癌细胞的增殖和迁移

在线全文

学校读者我要写书评

暂无评论

中国肿瘤生物治疗杂志 2020年第4期27卷 370-376页

作者：洪秋双王琳娄加陶上海交通大学附属胸科医院检验科上海200030

目的:探讨miR-93/Eph受体A4(EphA4)分子轴通过细胞外调节蛋白激酶(extracellular regulated protein kinases),ERK通路对非小细胞肺癌(non-small cell lung cancer,NSCLC)H460和H1299细胞增殖和迁移的影响。方法:用qPCR检测H460和H1299细胞中miR-93表达水平。分别在H460细胞中转染miR-93模拟物(mimics)和EphA4过表达质粒、在H1299细胞中转染miR-93抑制剂(inhibitor)后,用MTT、Transwell实验检测miR-93对转染细胞增殖和迁移的影响。用双荧光素酶报告基因实验验证miR-93与EphA4之间的靶向调控关系。用Western blotting检测细胞中增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、EphA4、ERK和p-ERK蛋白的表达水平,用MTT、Transwell实验检测同时过表达miR-93和EphA4对H460细胞增殖和迁移的影响。结果:miR-93在H1299细胞中表达水平高于H460细胞(P<0.01)。过表达miR-93促进H460细胞增殖和迁移(均P<0.01),敲低miR-93抑制H1299细胞增殖和迁移(均P<0.01)。双荧光素酶报告基因实验证实miR-93靶向调控EphA4,过表达miR-93明显下调H460细胞中EphA4 mRNA和蛋白表达水平(均P<0.05),过表达miR-93通过靶向EphA4并激活ERK通路促进H460细胞的增殖和迁移(均P<0.01)。结论:miR-93促进NSCLC细胞增殖和迁移,其机制可能与靶向调控EphA4并激活ERK通路有关。

关键词：非小细胞肺癌 H460细胞 H1299细胞 miR-93 EphA4 ERK通路增殖迁移

循环肿瘤细胞在良恶性肺部孤立性结节鉴别诊断中的应用价值

在线全文

学校读者我要写书评

暂无评论

中华检验医学杂志 2016年第12期39卷 941-945页

作者：乔理华王琳娄加陶上海交通大学附属胸科医院检验科 200030

目的：研究外周血中循环肿瘤细胞（ CTC）检测在肺部孤立性结节鉴别诊断中的价值。方法选取上海市胸科医院2013年9月至2015年1月134例因肺部孤立性结节入院患者，其中恶性结节患者80例，良性结节患者54例，采用配体靶向的聚合酶链反应（... 详细信息

目的：研究外周血中循环肿瘤细胞（ CTC）检测在肺部孤立性结节鉴别诊断中的价值。方法选取上海市胸科医院2013年9月至2015年1月134例因肺部孤立性结节入院患者，其中恶性结节患者80例，良性结节患者54例，采用配体靶向的聚合酶链反应（ LT－PCR）方法检测患者3 ml外周血中CTC数量，采用流式荧光法检测入组者血清中CEA和细胞角质蛋白19片段（CYFRA21－1）的表达水平，通过与肿瘤标志物癌胚抗原（ CEA）和CYFRA21－1做比对，评价外周血中CTC水平在良恶性肺部孤立性结节鉴别诊断中的价值。结果采用Mann－Whitney U检验，得出肺部恶性结节患者外周血中CTC数量[11．06（8．77～14．41） units／3 ml]显著高于良性结节[6．65（4．49～7．84） units／3 ml]患者，差异有统计学意义（Z＝－6．217，P＜0．001）；在肺部孤立性结节鉴别诊断中，CTC的敏感度为80％（64／80），特异度为85％（46／54）。根据结节直径大小进行统计，CTC对直径小于8 mm的肺部孤立性结节诊断敏感度为6／9，特异度为4／5；对直径8～20 mm的肺部孤立性结节诊断敏感度为83％（35／42），特异度为85％（29／34）；对直径大于20 mm的肺部孤立性结节诊断敏感度为79％（23／29），特异度为13／15。结论相对于传统的肿瘤标志物，CTC对肺部孤立性结节有更好的鉴别诊断性能。

关键词：孤立性肺结节肿瘤细胞,循环诊断,鉴别