循环肿瘤细胞捕获方法的研究进展

血液中的循环肿瘤细胞（CTCs）携带着肿瘤组织的遗传和表型信息，是液体活检的重要标志物。监测和分析血液中CTCs的数量和性质对癌症的早期诊断、治疗方案的确定和疗效评估具有重要意义。然而CTCs在血液中的含量极低，实现对CTCs的捕获与检测极具挑战。该文综述了基于生物物理原理、生物亲和原理以及人工抗体的CTCs捕获方法，并从捕获效率、捕获纯度和释放活性保持等方面进行了评述。此外，该文还对CTCs捕获方法的发展趋势进行了展望。     

肿瘤外泌体的分析检测

肿瘤外泌体是由肿瘤细胞释放到细胞外环境的微小囊泡,其直径约为30~150 nm,主要存在于血液、尿液、唾液等多种体液中,是早期肿瘤诊断的标志物之一。肿瘤外泌体具有较好的稳定性且含量丰富,因此是液体活检标志物的研究热点。肿瘤外泌体携带母细胞相关的蛋白、脂质和核酸等生物活性物质,为生物检测提供了多种特征标志物。本文就肿瘤外泌体的生成、分离、表征及分析检测进行了论述,重点讨论了肿瘤外泌体检测的研究进展。     

循环肿瘤细胞及细胞外囊泡的纳米检测技术

肿瘤液体活检通过对体液中生物标志物的检测实现疾病的精准诊断,对于恶性肿瘤的早期诊断和动态监测至关重要。循环肿瘤细胞(CTC)是肿瘤组织释放到血液中的肿瘤细胞,细胞外囊泡(EV)是由细胞分泌的膜囊泡,二者都携带肿瘤分子信息,并与肿瘤进展和转移密切相关,是重要的液体活检生物标志物,并且在检测方法和临床意义方面也有很多共性。纳米材料由于其高比表面积、独特的光、电、磁等物理化学特性以及易于功能化修饰等特点,被广泛用于CTC和EV的检测,以提高检测灵敏度和特异性,提供肿瘤形成、进展、转移和治疗反应的信息,具有很好的应用前景。本文回顾了在CTC和EV的特异性识别、高效捕获或分离、目标CTC或EV的鉴定等三方面的纳米技术进展,包括提高分子识别特异性的纳米材料表面识别探针功能化修饰以及捕获、鉴定的纳米材料和纳米技术的最新进展,总结了基于功能化纳米材料的液体活检技术的优势及挑战,为液体活检纳米技术的发展提供信息。     

膀胱癌病人尿液中的特征物质研究

采用顶空气相色谱/质谱联用法分析膀胱癌病人的尿液,寻找尿液中的挥发性特征物质,以期作为尿液检查诊断膀胱癌的一种方式。选取一组膀胱癌患者,对其尿液进行定性定量分析,并将分析结果与健康人尿液进行比较。进样口温度280℃,进样量100μL;起始柱温45℃,保留3min,以16℃/min的速率程序升温到280℃。结果表明:膀胱癌病人尿液中含有两种特征物质4-庚酮和二甲基硫醚。膀胱癌患者尿液中4-庚酮的浓度范围为0.14～1.32μL/L;二甲基硫醚的浓度范围为0.009～0.73μL/L。推测4-庚酮可能来源于增塑剂邻苯二甲酸二异辛酯(di(2-ethylhexyl)phthalate,DEHP)在人体内的降解;二甲基硫醚来源于含硫化合物在人体内的合成与降解。尿液中4-庚酮和二甲基硫醚的含量与膀胱癌具有相关性。     

口腔癌相关唾液肿瘤生物标志物的分析检测研究进展

口腔癌系头颈部癌，癌组织均位于口腔内，其非侵入性早期诊断是减少该病死亡的有效手段.唾液系口腔癌变相关物质首先释放进入的体液，取材方便，安全无创，是口腔癌普查筛选、早期诊断的首选指标.本文对唾液肿瘤生物标志物的种类、目前国内外常用的检测方法进行概述，重点阐述新型电化学生物传感方法在口腔癌相关唾液肿瘤生物标志物检测方面的相关应用及其最新研究进展.并对口腔癌相关唾液肿瘤生物标志物电化学传感技术的未来发展方向提出展望，拟为其深入研究与应用提供参考.     

核酸适配体功能化纳米界面用于循环肿瘤细胞捕获与释放

循环肿瘤细胞(CTCs)是肿瘤研究和临床癌症诊断中的重要对象,也是"液体活检"的重要标志物.CTCs携带着肿瘤组织的遗传和表型信息,有助于肿瘤的早期诊断、个体化治疗和预后监测.然而,CTCs是一种极其罕见的细胞群体,在癌症患者外周血中十分稀少,这对从患者血液中分离CTCs并无损释放进行下游分析提出了挑战.目前,基于CT...     

血液中循环肿瘤细胞捕获与释放方法研究进展

血液中的循环肿瘤细胞(CTCs)与癌症转移密切相关,对CTCs进行检测有利于实现癌症早期诊断。但由于血液中CTCs个数稀少且存在异质性,因此急需发展血液中CTCs的高效分离及检测方法。此外,释放被捕获的细胞并进行后续培养及基因水平分析,将会进一步推进癌症的个性化治疗。本文综述了近年来CTCs捕获及释放的相关研究进展,并展望了其应用前景及发展方向。     

人体血液、尿液、唾液及呼出气体中乙醇含量检测研究进展

对人体血液、尿液、唾液及呼出气体中乙醇含量检测方法进行综述。顶空气相色谱法适用于检测血液和尿液中的乙醇含量，精度高，稳定性好；乙醇氧化酶法在检测乙醇含量时有操作简单使用方便的特点；利用电化学原理或红外原理检测呼出气体中酒精含量时，可以实现定性、定量分析，仪器操作简单、稳定性好、体积小。对乙醇检测仪器和装置的溯源方法进行总结，国家有证标准物质的研制以及乙醇气体发生源装置的建立确保了检测结果的量值准确和单位统一。     

人巨细胞病毒检测在婴幼儿肺炎中的应用

目的探讨HCMV检测在婴幼儿肺炎中的诊断价值和临床应用。方法对2013年1月—12月180例在呼伦贝尔市人民医院诊断为婴幼儿肺炎的患儿进行外周血HCMV-IgM和尿液HCMV-DNA的检测,分别采用酶联免疫捕获法(ELISA)、荧光定量聚合酶链反应(PCR)对收集的血清及尿液进行检测。结果 180例婴幼儿肺炎中确诊为HCMV感染的72例,其中54例血清HCMV-IgM检测阳性,阳性率占75%,62例尿液HCMV-DNA检测阳性,阳性率占86%。对检测出HCMV的患儿进行临床跟踪调查发现用更昔洛韦等药物治疗后疗效显著。结论婴幼儿肺炎中HCMV感染所占比率高,通过血清HCMV-IgM及尿液HCMV-DNA的联合检测能够给临床提供一个治疗依据,有助于早期诊断、早期治疗,对减少并发症及预后至关重要。     

基于纳米探针的肿瘤细胞电化学传感研究进展

纳米材料在纳米尺度下具有独特的光、电、磁等性质,已广泛应用于细胞电化学传感的研究,该研究工作提高了肿瘤细胞检测的灵敏度与特异性,为癌症的早期诊断与治疗提供了新的方法与思路。本文综述了基于纳米探针进行肿瘤细胞电化学传感研究的工作,总结了纳米电化学细胞传感技术在肿瘤细胞固定、识别、检测及诱导凋亡等方面的研究进展,并对未来研究动向进行了展望。     

Molecular genetic diagnosis of de novo and recurrent bladder cancer

Bladder cancer, the second most common urological tumor, is usually diagnosed by endoscopy and biopsy of the lower urinary tract. However, this procedure is expensive, can cause discomfort to the patient and is a source of infection. Commercially available diagnostic systems measure protein byproducts of bladder carcinoma in voided urine; their sensitivity is only between 60-80%. Polymerase chain reaction (PCR)-based microsatellite analysis of the urine sediment (MAUS) is a noninvasive, inexpensive and easily performed analytical method which was introduced in the de novo diagnosis and follow-up of bladder cancer. By utilizing the PCR with 20 polymorphic microsatellite markers on different chromosomes and separating PCR products by electrophoresis on 7% denaturing polyacrylamide-formamide-urea slab gels, a 91% diagnostic sensitivity could be achieved. In order to minimize costs and analysis time, the separation and detection of PCR products was carried out by capillary array electrophoresis and two-color fluorescent primer labeling/laser beam detection in another study. The accuracy of both methods was the same. In either detection system, MAUS is an accurate and promising tool in the noninvasive diagnosis of bladder cancer.     

Quantitative determination of taurine and related biomarkers in urine by liquid chromatography–tandem mass spectrometry

Current urinary bladder cancer diagnosis is commonly based on a biopsy obtained during cystoscopy. This invasive method causes discomfort and pain in patients. Recently, taurine and several other compounds such as L-phenylalanine and hippuric acid in urine were found to be indicators of bladder cancer. However, because of a lack of sensitive and accurate analytical techniques, it is impossible to detect these compounds in urine at low levels. In this study, using liquid chromatography–tandem mass spectrometry (LC-MS/MS), a noninvasive method was developed to separate and detect these compounds in urine. ¹⁵N₂-L-glutamine was used as the internal standard, and creatinine acted as an indicator for urine dilution. A phenyl-hexyl column was used for the separation at an isocratic condition of 0.2% formic acid in water and 0.2% formic acid in methanol. Analytes were detected in multiple-reaction monitoring with positive ionization mode. The limit of detection range is 0.18–6 nM and the limit of quantitation ranges from 0.6 to 17.6 nM. The parameters affecting separation and quantification were also investigated and optimized. Proper clinical validation of these biomarkers can be done using this reliable, fast, and simple method. Furthermore, with simple modifications, this method could be applied to other physiological fluids and other types of diseases.     

Bladder cancer hunting: A microfluidic paper-based analytical device

Bladder cancer is the fourth most common cancer in men, and it is becoming a prevalent malignancy. Most of the regular clinical examinations are prompt evaluations with cystoscopy, renal function testing, which require high-precision instrument, well-trained operators, and high cost. In this study, a microfluidic paper-based analytical device (μPAD) was fabricated to detect nuclear matrix protein 22 (NMP22) and bladder cancer antigen (BTA) from the urine samples. Urine samples were collected from 11 bladder cancer patients and 10 well-beings as experiment and control groups, respectively, to verify the working efficiency of μPAD. A remarkable checkout efficiency of up to 90.91% was found from the results. Meanwhile, this method is feasible for home-based self-detection from urine samples within 10 min for the total process, which provides a new way for quick, economical, and convenient tumor diagnosis, prognosis evaluation, and drug response.     

Photoelectrochemical genosensors for the determination of nucleic acid cancer biomarkers

Liquid biopsy consists of a test or a series of tests carried out on a sample of blood to monitor circulating biomarkers, including nucleic acids. In cancer, a liquid biopsy approach can be useful not only for a rapid diagnosis, but also for therapy selection and cancer patient monitoring during treatment. However, the analytical assays need to be very sensitive (capable of detecting very small amounts of circulating analytes) as well as selective.Photoelectrochemistry is a powerful technique of developing an analytical biosensing assay, since it possesses advantages of both optical and electrochemical methods by coupling light excitation and electrochemical detection, and thus resulting in potentially very sensitive measurements due to the low background current. The aim of this review is to describe photoelectrochemical genosensors and to point out the potential of this class of genosensors in determining circulating nucleic acids as well as the challenges in the frame of a liquid biopsy approach.     

Proteomic profiling of human urinary proteome using nano-high performance liquid chromatography/electrospray ionization tandem mass spectrometry

Urine, a blood filtrate produced by the urinary system, is an ideal bio-sample and a rich source of biomarkers for diagnostic information. Many components in urine are useful in clinical diagnosis, and urinary proteins can be strong indication for many diseases such as proteinuria, kidney, bladder and urinary tract diseases. To enhance our understanding of urinary proteome, the urine proteins were prepared by different sample cleanup preparation methods and identified by nano-high performance liquid chromatography electrospray ionization tandem mass spectrometry followed by peptide fragmentation pattern. The experimental results demonstrated that a total of 2283 peptides, corresponding to 311 unique proteins, were identified from human urine samples, in which 104 proteins with higher confidence levels. The present study was designed to establish optimal techniques to create a proteomic map of normal urinary proteins. Also, a discussion of novel approaches to urine protein cleanup and constituents is given.     

Design of high stability thin-film transistor biosensor for the diagnosis of bladder cancer

Bladder cancer is the most common malignant tumor in the urinary system,with high morbidity,mortality and recurrence after surgery.However,current bladder cancer urine diagnosis methods are limited by the low accuracy and specificity due to the low abundance of bladder cancer biomarkers in the urine with complex biological environments.Herein,we present a high stability indium gallium zinc oxide field effect transistor(IGZO-FET) biosensor for efficient identification of bladder cancer biomarkers from human urine samples.The recognition molecular functionalized IGZO-FET biosensor exhibits stable electronic and sensing performance due to the large-area fabrication of IGZO thin-film FET.Owing to the excellent electrical performance of IGZO-FET,the IGZO-FET biosensor exhibits high sensitivity and extremely low detection limit(2.7 amol/L) towards bladder cancer biomarkers.The IGZO-FET biosensor is also able to directly detect bladder tumor biomarker in human urine with high sensitivity and specificity,and could differentiate bladder cancer patients' urine samples from healthy donors effectively.These results indicate that our designed high-performance biosensor shows great potential in the application of portable digital bladder cancer diagnosis devices.     

Construction of high stability indium gallium zinc oxide transistor biosensors for reliable detection of bladder cancer-associated microRNA

Bladder cancer is the most common malignant tumours with high morbidity, mortality and recurrence.However, currently developed detection methods for bladder cancer-associated urine biomarkers are hindered by their extremely low abundance. Hence, the exploration of a highly sensitive and selective approach for the detection of trace bladder cancer-associated biomarkers in human urine is of vital importance for the diagnosis of bladder cancer. Herein, we developed a highly reliable indium gallium ...     

尿核基质蛋白-22和液基细胞学在血尿患者膀胱癌筛查中的应用

目的探讨尿核基质蛋白-22和液基细胞学在血尿患者膀胱癌筛查中的应用。方法选取2015年6月至12月在兴宁市中医医院收集的10例膀胱癌疑似患者,分别进行尿核基质蛋白-22检查与尿液基细胞学检。观察两种检查下患者的诊断结果。结果尿核基质蛋白-22的敏感度为80.2%,液基细胞学的敏感度为72.1%,两组之间作比较差异有统计学意义(P0.05)。尿核基质蛋白的特异度为71.6%,液基细胞学的特异度为98.6%,两组之间比较差异具有统计学意义(P0.05)。两种检查下患者阳性预测率及阴性预测率比较,差异显著。结论在血尿患者的膀胱癌筛查中,尿核基质蛋白22作为诊断膀胱癌的新肿瘤标记物对患者创伤性较小,灵敏度和特异性较高的诊断方法。而尿液基细胞学与核基质蛋白22联合使用可以有效提高对膀胱癌诊断的敏感程度。     

Evaluation of a gas sensor array and pattern recognition for the identification of bladder cancer from urine headspace

Previous studies have indicated that volatile compounds specific to bladder cancer may exist in urine headspace, raising the possibility that headspace analysis could be used for diagnosis of this particular cancer. In this paper, we evaluate the use of a commercially available gas sensor array coupled with a specifically designed pattern recognition algorithm for this purpose. The best diagnostic performance that we were able to obtain with independent test data provided by healthy volunteers and bladder cancer patients was 70% overall accuracy (70% sensitivity and 70% specificity). When the data of patients suffering from other non-cancerous urological diseases were added to those of the healthy controls, the classification accuracy fell to 65% with 60% sensitivity and 67% specificity. While this is not sufficient for a diagnostic test, it is significantly better than random chance, leading us to conclude that there is useful information in the urine headspace but that a more informative analytical technique, such as mass spectrometry, is required if this is to be exploited fully.