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1.
贺晖  周玲俐  刘震 《化学学报》2021,79(1):45-57
异常的蛋白质表达与疾病的发生与发展密切相关, 因此蛋白质已作为疾病标志物广泛应用于疾病的早期诊断、治疗监测和预后评估. 然而, 临床样本中的蛋白质疾病标志物通常含量极低, 并存在高丰度的基质干扰, 对检测方法的特异性和灵敏度提出挑战. 目前, 蛋白质疾病标志物的检测方法主要是免疫分析. 但是, 免疫分析主要依赖抗体进行特异性识别, 而抗体具有不易制备、稳定性较差和成本高等缺点. 同时, 免疫分析常通过荧光和化学发光等技术实现高灵敏检测, 但存在操作繁琐、光漂白、光谱宽等不足. 分子印迹聚合物已发展成为在特异性和亲和力方面可媲美抗体的仿生识别材料, 且具有容易制备、稳定性好和成本低等优势. 表面增强拉曼散射技术具有超高灵敏度、光谱窄、快速、无损检测等优势而广泛应用于化学和生物分析. 近年来, 分子印迹技术和表面增强拉曼散射技术的结合产生了系列先进的蛋白质检测方法, 展现了独特的优势, 受到了广泛的关注. 本综述旨在介绍该联用分析技术的主要进展, 在分别介绍分子印迹和表面增强拉曼散射及其在蛋白质检测中单独应用的基础上, 着重介绍基于两种技术的蛋白质疾病标志物的检测方法的研究进展. 最后, 对该联用技术的未来发展做了展望.  相似文献   
2.
Lipid metabolism has a significant function in the central nervous system and Alzheimer's disease (AD) is an age-related senile disease characterized by central nerve degeneration. The pathological development of AD is closely related to lipid metabolism disorders. To reveal the influence of Kai-Xin-San (KXS) on lipid metabolism in APP/PSI transgenic mice and potential therapeutic targets for treating AD, brain tissue samples were collected and analyzed by high-throughput lipidomics based on UPLC–Q/TOF-MS. The collected raw data were processed by multivariate data analysis to discover the potential biomarkers and lipid metabolic profiles. Compared with the control wild-type mouse group, nine potential lipid biomarkers were found in the AD model group, of which seven were up-regulated and two were down-regulated. Orally administrated KXS can reverse the changes in these potential biomarkers. Compared with the model group, a total of six differential metabolites showed a recovery trend and may be potential targets for KXS to treat AD. This study showed that high-throughput lipidomics can be used to discover the perturbed pathways and lipid biomarkers as potential targets to reveal the therapeutic effects of KXS.  相似文献   
3.
采用顶空固相微萃取(HS-SPME)与气相色谱-质谱(GC-MS)联用技术,对12例卵巢癌、17例成熟型畸胎瘤与16例正常血液样本的挥发性组分进行研究,对影响提取效果的实验因素进行了优化.在最优条件下共检出28种挥发性组分,以各组分峰面积为变量,用SIMCA-P软件进行偏最小二乘判别分析(PLS-DA),3组样本被明显区分.同时,依据PLS-DA载荷图得到8种具有显著性差异的标志物,其中己醛、蘑菇醇的变化与其在肝癌、肺癌患者血液中的变化一致,可作为诊断卵巢癌和成熟型畸胎瘤的挥发性生物标志物.  相似文献   
4.
In the present study, the voltammetric and impidimetric detection of microRNA‐21, mir‐21 from cell lysates was investigated for the first time by using graphene modified disposable pencil graphite electrodes (GME). The surface characterization of GME was performed via electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). Upon passive adsorption of inosine substituted antimicroRNA‐21, antimir‐21 probe, InP, onto the surface of GME and then solid phase hybridization of InP with mir‐21, the target, the electrochemical detection was performed by using Differential Pulse Voltammetry (DPV) and EIS techniques. This developed biosensor, GME has presented a 2.77 times lower detection limit of 2.09 µg/mL (3.12 pmol) with respect to unmodified pencil graphite electrode (GE). Moreover it is capable of analyzing mir‐21 in the cell lysates of mir‐21 positive breast cancer cell line (MCF‐7) contrast to mir‐21 negative hepatoma cell line (HUH‐7). The proposed electrochemical yes‐no system does not require any purification and/or amplification step prior to fast detection of mir‐21 from real samples.  相似文献   
5.
广西合山石村矿超高有机硫煤饱和烃特征分析   总被引:1,自引:0,他引:1  
以合山石村矿超高有机硫煤为研究对象,利用带能谱的扫描电镜(SEM-EDX)及气相色谱(GC)和色谱-质谱联用(GC-MS)分析技术,综合分析探讨了研究区样品中有机质来源、沉积环境及物源输入。结果表明,研究区样品镜质体反射率均值1.76%,全硫含量均值6.01%,其中有机硫含量占比达94.3%,为高成熟度的超高有机硫煤。饱和烃色谱图中未分辨的复杂混合物(UCM)鼓包明显,前峰及双峰型均有分布,前峰型主峰碳为C_(16)、C_(18)、C_(21),双峰型主峰碳为C_(18)、C_(27),化合物降姥鲛烷、脱氢松香烷、咔达烯均有发现,规则甾烷C_(27)、C_(28)、C_(29)均呈"V"型分布,说明沉积母质受藻类等低等水生生物与高等植物双重输入的影响。姥植比参数、C_(31-35)藿烷呈阶梯式递减的分布特征表明在海相碳酸盐台地成煤带中存在一定的氧化条件。扫描电镜中拍摄到的铁含氧硫酸盐、细胞充填环状黄铁矿说明在沉积质晚期成岩阶段受到一定程度热液作用影响。  相似文献   
6.
The development of early and personalized diagnostic protocol with rapid response and high accuracy is considered the most promising avenue to advance point-of-care testing for tumor diagnosis and therapy. Given the growing awareness of the limitations of conventional tissue biopsy for gathering tumor information, considerable interest has recently been aroused in liquid biopsy. Among a myriad of analytical approaches proposed for liquid biopsy, microfluidics-based separation and purification techniques possess merits of high throughput, low samples consumption, high flexibility, low cost, high sensitivity, automation capability and enhanced spatio-temporal control. These characteristics endow microfluidics to serve as an emerging and promising tool in tumor diagnosis and prognosis by identifying specific circulating tumor biomarkers. In this review, we will put our focus on three key categories of circulating tumor biomarkers, namely, circulating tumor cells (CTCs), circulating exosomes, and circulating nucleic acids (cNAs), and discuss the significant roles of microfluidics in the separation and analysis of circulating tumor biomarkers. Recent advances in microfluidic separation and analysis of CTCs, exosomes, and cNAs will be highlighted and tabulated. Finally, the current challenges and future niches of using microfluidic techniques in the separation and analysis of circulating tumor biomarkers will be discussed.  相似文献   
7.
Human Mammary Tumor Virus (HMTV) or Mouse Mammary Tumor Virus holds similarity as an endogenous onco-retrovirus belongs to retroviridae family, predominantly infects the epithelial cell of human as well as mouse. With the recognition of nano-biosensor in nanotechnology, ideal interdigitated electrode (IDE) was genuinely performed for HMTV detection. Aluminium enriched IDE (AlIDE) was fabricated for high performance detection with a cost-effective photolithography technique. In this research, (3-glycidyloxypropyl) trimethoxysilane refined platform was selected to detect the conductivity with HMTV target DNA interaction on the designed AlIDE. Strong binding affinity of streptavidin-biotin with target DNA enhanced the sensitivity by empowering higher number of HMTV probe and target complementation on sensing surface. Furthermore, the target DNA was immobilized on probe modified AlIDE and a quantitative value of 100 aM attained as a lowest detection. A linear with dose-dependent duplex formation was shown with the regression coefficient value of 0.964. Negative control has shown insignificant detection at 10 pM, which justifies the higher fold discrimination with specificity. The excellence of AlIDE performance in detection of HMTV may pave the way for more verification on other diseases.  相似文献   
8.
《中国化学》2018,36(1):42-46
Two rosamine‐based pH probes 1a and 1b with pyronine‐phenol skeleton were designed and synthesized by a simple one‐step reaction. pH titration experiments showed that probes 1a and 1b exhibit near OFF–ON fluorescence responses around 550–750 nm towards the hydrogen ions. The pKa of the probe 1a is 8.29, while that of the probe 1b increases to 12.1 because of the hydrogen bond inside it. Selective and competitive experiments indicated that both common ions and amino acids did not interfere their emission with hydrogen ions. Moreover, confocal fluorescent imaging showed that the probe 1a could be served as mitochondria biomarker in HeLa and Ges‐1 cells.  相似文献   
9.
Serum levels of fully sialylated C4‐binding protein (FS‐C4BP) are remarkably elevated in patients with epithelial ovarian cancer (EOC) and can be used as a marker to distinguish ovarian clear cell carcinoma from endometrioma. This study aimed to develop a stable, robust and reliable liquid chromatography–hybrid mass spectrometry (UPLC‐MS/MS) based diagnostic method that would generalize FS‐C4BP as a clinical EOC biomarker. Glycopeptides derived from 20 μL of trypsin‐digested serum glycoprotein were analyzed via UPLC equipped with an electrospray ionization time‐of‐flight mass spectrometer. This UPLC‐MS/MS‐based diagnostic method was optimized for FS‐C4BP and validated using sera from 119 patients with EOC and 127 women without cancer. A1958 (C4BP peptide with two fully sialylated biantennary glycans) was selected as a marker of FS‐C4BP because its level in serum was highest among FS‐C4BP family members. Preparation and UPLC‐MS/MS were optimized for A1958, and performance and robustness were significantly improved relative to our previous method. An area under the curve analysis of the FS‐C4BP index receiver operating characteristic curve revealed that the ratio between A1958 and A1813 (C4BP peptide with two partially sialylated biantennary glycans) reached 85%. A combination of the FS‐C4BP index and carbohydrate antigen‐125 levels further enhanced the sensitivity and specificity.  相似文献   
10.
The identification of biomarkers would be of benefit for the diagnosis and treatment of colorectal cancer. DNA methylation in specific genomic regions, which had shown strongly association with disease genotypes, was an effective indicator to reveal the occurrence and development of cancers. To screen out methylation biomarkers for colorectal cancer (CRC), genomic DNA was isolated from colorectal cancerous and corresponding cancer‐adjacent tissues collected from 30 CRC patients and then bisulfite‐converted. The exon regions of 5 targeted genes (CNRIP1 , HIC1 , RUNX3 , p15 , and SFRP2 ) were amplified by using nested polymerase chain reaction with specific primers, and the amplicon was purified and hydrolyzed. The methylation levels of these specific regions were detected by liquid chromatography tandem mass spectrometry (LC‐MS/MS). The results showed that 5 targeted exon regions were successfully amplified and confirmed by sequencing. The methodological validations indicated that LC‐MS/MS was highly sensitive and accurate. The methylation levels of CNRIP1 and RUNX3 were remarkably high in CRC tissues with statistical difference when compared with corresponding cancer‐adjacent individuals, while that of HIC1 , p15 , and SFRP2 had no difference between 2 subjects. These findings supported CNRIP1 and RUNX3 as potential DNA methylation biomarkers for CRC diagnosis and treatment, and our LC‐MS/MS approach exhibited great advantages in the identification of regional DNA methylation biomarkers.  相似文献   
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