激光诱发肿瘤荧光图像定位方法的研究进展

激光诱发肿瘤荧光图像定位方法是利用光敏剂在癌组织中的选择聚集性和受激发射荧光特性,通过实时检测荧光图像,可较为准确地识别恶变肿瘤组织的范围,能在肿瘤手术过程中对肿瘤图像进行实时显示,有效减小手术或其它治疗对患者重要器官的损伤;此外,在早期诊断癌症的过程中,这种方法可以引导组织活检,并为X射线检查提供辅助信息,对早期癌变组织进行诊断和定位。     

乳腺癌分子分型用荧光分子影像探针

目前,乳腺癌的临床诊断就是结合病理类型和分子分型 (免疫组化)。该方法是有创的,而且不能原位、实时地展现关键生物分子与临床关键信息之间的关系。本文介绍了利用分子影像技术对乳腺癌进行分子分型检测的最新研究进展。荧光成像灵敏度高,且不过分依赖图像分析,只需相应的荧光分子探针,即可实时、定量或半定量、多通道地得到肿瘤组织分子分型信息。因此,研制安全高效、组织穿透力强的近红外荧光分子探针是未来荧光成像技术和乳腺癌分子分型研究的重点。     

光敏剂浓度和给药时间对荧光诊断效果的影响

目的 为探索光敏剂浓度和给药时间对蓝光诱导的荧光诊断的影响,以提高荧光诊断的准确度.方法 常规方法制备荷瘤小鼠,分别给荷瘤小鼠静脉注射不同浓度（0～70 mg/kg体质量）的光敏剂-血卟啉衍生物（HpD）,并在给药后不同时间（0～48 h）,以前置395～415 nm滤光片的碘镓灯为激发光源,蓝光诱导载体荧光,通过图像采集系统采集荧光图像,取活组织做病理切片检查.结果 HpD浓度为40mg/kg体质量,给药后18 h,可以较为精确地反映肿瘤组织的浸润范围,荧光诊断准确度高于其他实验组;同时假阳性率低于其它实验组,差异有统计学意义.结论 荧光诊断的效果与光敏剂的浓度和观察时间密切相关;优化2者参数,可提高荧光诊断的准确度并降低假阳性率,对临床肿瘤手术方案的制定有重要参考价值.     

激光诱发荧光在结肠肿瘤早期诊断中的应用

激光诱导荧光法（ＬＩＦ）利用生物组织的自体荧光特性来判断组织性质，能实时，无损地提供组织信息，从而区分正常与病变组织。本文详述了生物组织自体荧光谱技术应用于结肠肿瘤诊断的研究方法，最新成果和现状，并对其发展和应用提供了几点建议。     

近红外荧光散射断层成像的研究进展

近红外荧光光学断层成像(FODT)是以合适的荧光探针作为标记物或对比剂，用特定波长的红光激发荧光染料，使其发出波长长于激发光的近红外荧光，通过测量媒质边界处有限点的荧光强度，考虑光子在组织中传播的散射特性，来重建出组织内部的荧光光学特性的分布图像以及组织光学参数。这种成像方式具有无电离辐射、染料稳定、可长期监测和设备简单、成本低等优点，在肿瘤检测、基因表达、蛋白质分子检测和药物受体定位等方面有着很大的应用潜力。在给出近红外荧光散射断层成像典型系统的基础上，详述了近红外荧光在组织中的频域传播模型和重建算法；介绍了两家研究机构在此领域的研究进展；讨论了将该成像方法应用于临床的进一步的发展方向。     

激光诱发荧光在结肠肿瘤早期诊断中的应用

激光诱导荧光法 (L IF)利用生物组织的自体荧光特性来判断组织性质 ,能实时、无损地提供组织信息 ,从而区分正常与病变组织。本文详述了生物组织自体荧光谱技术应用于结肠肿瘤诊断的研究方法、最新成果和现状 ,并对其发展和应用提出了几点建议。     

近红外荧光散射断层成像的研究进展

近红外荧光光学断层成像(FODT)是以合适的荧光探针作为标记物或对比剂,用特定波长的红光激发荧光染料,使其发出波长长于激发光的近红外荧光,通过测量媒质边界处有限点的荧光强度,考虑光子在组织中传播的散射特性,来重建出组织内部的荧光光学特性的分布图像以及组织光学参数。这种成像方式具有无电离辐射、染料稳定、可长期监测和设备简单、成本低等优点,在肿瘤检测、基因表达、蛋白质分子检测和药物受体定位等方面有着很大的应用潜力。在给出近红外荧光散射断层成像典型系统的基础上,详述了近红外荧光在组织中的频域传播模型和重建算法;介绍了两家研究机构在此领域的研究进展;讨论了将该成像方法应用于临床的进一步的发展方向。     

可见光和近红外荧光分光融合成像外科手术导航系统的研制

本文提出研制一种在开放手术过程中无创、实时、定位的外科手术光学导航系统。系统设计依据近红外荧光分子成像原理,采用活体荧光激发技术以及多通道分光摄像技术和图像融合软件技术。研制过程中将可见光和近红外光环形LED激发光源、多通道带通滤光片、分光摄像2CCD光电传感器和计算机系统技术集成,成功研制出一种新型外科手术光学导航系统。该系统在活体注射近红外荧光显影剂时,能够显示同一部位、同一时刻手术野组织表面的解剖学图像和近红外荧光功能图像。解决了医生在术中肉眼难以识别淋巴管、淋巴结、肿瘤边缘的技术问题,实现实时、无创的外科手术导航,有效引导外科医生切除肿瘤组织,显著提高手术成功率。该项技术已获得国家专利,专利号为ZI.2011 1 0292374.1。     

荧光原位杂交在软组织肿瘤石蜡包埋组织中的应用

荧光原位杂交（fluorescence in situ hybridization，FISH）作为一种染色体或基因评价的有力工具，可将基因缺失、扩增、异位等异常与组织形态学结合起来，提高病理诊断的准确性，尤其对一些具有遗传学异常而组织形态不典型的软组织肿瘤的诊断。我们采用FISH的方法检测韧带样型纤维瘤病中8号染色体的数目异常，探讨软组织肿瘤石蜡包埋组织FISH检测的可行性、方法优化及其应用。     

量子点探针辅助的近红外荧光成像技术在肿瘤显影中的应用

肿瘤早期检测是精准并高效诊疗癌症的关键因素。荧光成像技术凭借其高灵敏度、高时空分辨率、无电离辐射和无创实时成像等优点,在生物医学领域,尤其在肿瘤检测方面展现出了广泛的应用前景。近红外光穿过生物组织时,受到的吸收和散射较少,因此在生物成像方面体现了高信噪比和强组织穿透能力。在众多荧光探针中,近红外发光的量子点探针因其量子产率高、抗光漂白性强、发射光可调和稳定性良好等特点在荧光成像方面显示出突出的优势。本文基于量子点探针的近红外荧光成像技术在肿瘤显影中的应用,介绍了量子点优异的光学性能,并重点讨论了硫化铅（PbS）和硫化银（Ag2S）近红外发光量子点探针在肿瘤成像方面的研究进展,并对近红外发光量子点探针的应用前景进行了展望。     

实时荧光定量PCR快速分子诊断唐氏综合征

目的　探讨快速诊断唐氏综合征的检测方法 .方法　采用实时荧光定量PCR技术 ,检测唐氏患者 2 3例、正常人 33例 ,定量分析比较ΔCt值 .结果　正常组和唐氏组ΔCt值有显著性差异 (p <0 .0 0 1) ,并建立了相应的参考值范围 .结论　实时荧光定量PCR具有简单、快速、特异性高等优点 ,可用于快速诊断唐氏综合征     

锌离子在小鼠肺组织分布的TSQ荧光研究

目的研究游离锌离子在小鼠肺组织的定位分布。方法应用TSQ荧光技术检测小鼠肺组织内的锌离子。结果荧光显微镜下可见被TSQ荧光标记的锌离子广泛的分布在肺叶支气管壁粘膜、粘膜下及软骨,肺内小支气管基膜,肺泡上皮和肺内血管平滑肌,尤其以肺叶支气管粘膜以及软骨中荧光染色最为明显。结论小鼠肺组织内含有丰富的锌离子可能参与肺泡上皮细胞以及肺各级支气管软骨细胞中某些蛋白质的合成和组装。     

Fluorescence polarization assay by flow cytometry

Fluorescence polarization measurement on cell suspensions provides a highly sensitive means for detecting subtle changes in the cells, such as occur early after lymphocyte activation or on malignant transformation. We review here the principles of fluorescence polarization, its measurement by a commercially available flow cytometer and application of such assays especially in cellular immunology.     

Finely Tuned Fluorescence Emission of Polydiphenylacetylene Films Obtained by Copolymerization

PDPAs containing longer (octadecyl, C18) and shorter (methyl, C1) alkyl side chains in the same backbone are synthesized. The copolymerization of C1 and C18 monomers at a range of feed ratios afford high‐molecular‐weight (M _w > 10⁶) copolymers almost quantitatively. As the content of longer alkyl chain units in the copolymers increases, the emission band in the bulk film shifts to shorter wavelengths and the emission intensity increases. The lamellar layer distance increases with increasing C18 content. The excited species with shorter lifetimes decrease gradually with increasing C18 content, whereas the longer‐lasting emission increases. Fine tuning of the FL emission of PDPAs is achieved by simple copolymerization of monomers with different alkyl side‐chain lengths.     

Mechanism of Protective Effect of Energostim during Catalepsy Produced by Single Treatment with Trifluoperazine

NAD, cytochrome c, and energostim modulated the fluorescence emission spectrum of trifluoperazine in the solution and in microsomal suspension. The data suggest that NAD and energostim modify structural and conformational characteristics of the dopamine receptor-trifluoperazine complex. These changes probably underlie the anticataleptic effect of energostim.     

Identification of mycobacteria to the species level by automated restriction enzyme fragment length polymorphism analysis

An automated method for the restriction fragment length polymorphism (RFLP) analysis for the differentiation of mycobacteria to the species level is described. After polymerase chain reaction (PCR) amplification of a sequence of the gene encoding the 65-kDa surface antigen common to all mycobacteria the product was investigated by RFLP analysis. For accurate determination of fragment sizes the asymmetrically fluorescein-labelled PCR product was partially digested with restriction site enzymes BstEII and HaeIII. The fragments obtained were analysed electrophoretically using an automated laser fluorescence DNA sequencer. Determination of fragment sizes revealed a deviation of ±1 base pair (bp; 0.6%) when compared to expected sizes. The validity of this approach was confirmed by analysing mycobacterial DNA obtained from pure cultures of Mycobacterium (M.) tuberculosis and alcohol-fixed smears as well as paraffin-embedded sputa of patients with culture-proven tuberculosis. Additionally a diagnostic algorithm was established by investigation of cultured M. bovis, M. bovis bacille Calmette-Guérin, M. avium, M. intracellulare and M. fortuitum. The method allows the identification of restriction enzyme sites which are only 40 bp apart. Partial restriction enzyme digestion of asymmetrically fluorescence-labelled PCR products will presumably lead to the discovery of new restriction enzyme sites.     

实时荧光定量RT-PCR法定量检测手足口病病原体的探讨

目的了解北京市门头沟区手足口病患儿感染病毒的型别,探索用荧光定量RT—PCR法对手足口病患者咽拭子标本中肠道病毒71型（EV71）、柯萨奇病毒A16（CoxA16）型病毒载量进行定量检测的可行性。方法采用实时荧光RT—PCR体外扩增法对81例手足口病患儿咽拭子标本提取的RNA进行检测。结果28例手足口病患者体内CoxA16病毒载量〉10^3 copies·ml-1。实时荧光定量RT—PCR法构建的标准曲线显示,样本阈值环数（Ct）值与病毒拷贝数的对数（10g10）之间的相关系数为0．9998,相关性良好。4例手足口病患者咽拭子标本中EV71型病毒载量〉10^3 copies·ml-1。实时荧光定量RT-PCR法标准曲线显示,Ct值与病毒拷贝数的对数之间的相关系数为0．9996,相关性较好。结论门头沟区手足口病病原谱以CoxA16型为主,34．6％的手足口病患儿CoxA16型病毒载量〉10^3 copies·ml-1,4．9％的患儿EV71型〉10^3 copies·ml-1。实时荧光定量RT-PCR法对咽拭子标本中CoxA16、EV71核酸定量检测比较简便快速、结果直观、稳定性强,为进一步探讨患者体内病毒载量与临床症状的关系打下了良好的基础。     

Determination of the Wall Thickness of Block Copolymer Vesicles by Fluorescence Lifetime Imaging Microscopy

This study reports on the characterization of reporter dye‐loaded block copolymer vesicles (polymersomes) of PS₁₁₅‐b‐PAA₁₅ (polystyrene‐block‐poly(acrylic acid)) and PEG₁₁₄‐b‐PLA₁₆₇ (poly(ethylene glycol)‐block‐poly(lactic acid)) and their drying behavior by fluorescence lifetime imaging microscopy (FLIM). The characteristic changes of the fluorescence decay components of the dye calcein incorporated in the three different local nanoenvironments, namely, the solvated dye, dye associated with the vesicle wall, and dried agglomerated dye, are observed by FLIM during the drying of vesicles. The amplitude ratio R _1/2 of the components attributed to the solvated dye in the vesicle interior with a lifetime τ₁ ≈ 3.9 ns to the one attributed to calcein associated with the wall with a lifetime τ₂ ≈ 1.5 ns is found to decrease exponentially with drying time. The time constants, which are found to depend linearly on the radius of the vesicle, yield by extrapolation to zero vesicle diameter an estimate of the polymersome wall thickness. For PS₁₁₅‐b‐PAA₁₅ and PEG₁₁₄‐b‐PLA₁₆₇ vesicle wall thicknesses r ₀ of 16 ± 5 nm and 28 ± 4 nm, respectively, are observed, which are in favorable agreement with transmission electron and atomic force microscopy data.

     

Synthesized of glucose-responsive nanogels labeled with fluorescence molecule based on phenylboronic acid by RAFT polymerization

We reported on the fabrication of sugar-responsive nanogels covalently incorporated with 3-acrylamidophenylboronic acid (AAPBA) as glucose-recognizing moiety, 2-(acrylamido)glucopyranose (AGA) as biocompatible moiety, and boron dipyrromethene (BODIPYMA) as fluorescence donor molecule. The p(AAPBA-AGA-BODIPYMA) nanogels were synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization in the mixture solvents of H₂O/ethanol. Nanogels could respond to glucose and size of nanogels increased after treating with 3?mg/mL glucose medium. The fluorescent intensity of nanogels varied dependent on different glucose concentrations. Besides, insulin, a model drug, can be encapsulated into nanogels with the loading amount up to 8.2%. The drug release was dependent on the content of AAPBA moieties in nanogels and glucose concentrations in release medium. The investigation on the cytotoxicity of nanogels revealed that nanogels had good compatibility. Such glucose-responsive nanogels have potential in detection and treatment of diabetes.