新型Gd基T2造影剂的制备和应用

设计并合成了结构为TPP-Lys（Acp-DOTA-Gd）-COOH（简称Gd-DOTA-TPP）的小分子磁共振探针,通过电转染的方式用探针标记人源脐带间充质干细胞（hMSCs）.11.7 T磁共振成像（MRI）扫描结果表明,Gd-DOTA-TPP标记的hMSCs在细胞内Gd含量为9×10⁹ Gd/cell时,T₂加权信号强度即可低至背景信号强度,呈现较强暗信号.将Gd-DOTA-TPP标记的hMSCs移植入小鼠脑室,可明显提高移植干细胞在MRI设备上的检测灵敏度,检测限可低至10³个细胞.     

胶束型磁共振成像分子探针的设计与应用

飞速发展的分子影像学在肿瘤的早期诊断及检测中发挥着越来越重要的作用.磁共振成像(MRI)是分子影像学的重要分支,具有其他成像技术不可比拟的优越性和广阔的发展前景.它不需要放射性示踪剂,没有电离辐射,具有高的空间、时间分辨率和组织对比度.近年来,新型磁共振分子探针及成像序列取得了一系列进展,包括环境响应型分子探针、¹⁹F成像、¹²⁹Xe超极化成像以及化学交换饱和转移成像等,进一步拓展了MRI的应用范围.研究和开发靶向性好、弛豫效率高且安全性好的新型多模态MRI造影剂,进一步提高灵敏度是MRI领域的一项重要课题,例如将胶束的特性与一些MRI新方法结合,寻找合适的胶束体系,以提高MRI分子探针的灵敏度;或者引入多模态分子探针,弥补磁共振方法的不足.本文综述了胶束型MRI分子探针核心技术的研究进展与应用,并指出分子影像技术在生物医学工程研究和临床诊断中的重要性.     

MRI与MRS在肥胖症研究中的应用进展

肥胖症已经成为严重威胁人类健康的主要慢性疾病之一,磁共振成像(MRI)和磁共振波谱(MRS)技术的结合运用,对于评价脂肪组织分布和蓄积程度具有极大的优势.该文总结了近年来MRI与MRS技术在肥胖症研究中的应用进展,并讨论了MRI与MRS技术在肥胖症临床应用及科学研究中的价值.     

STZ诱导大鼠1型糖尿病进行性脑萎缩的磁共振成像及组织化学研究

1型糖尿病(T1DM)是一种慢性代谢疾病,主要表现为胰岛素分泌量较正常情况下降,会对人体的多个器官和系统造成持续性的损伤．关于糖尿病的横向研究发现糖尿病患者相比于正常人存在着显著的脑萎缩,但关于糖尿病引起的脑萎缩随时间发生进行性改变的研究比较少见．实验采用腹腔注射链脲佐菌素(STZ)来诱导建立大鼠的1型糖尿病模型,运用磁共振成像(MRI)的方法对萎缩的脑区进行定位并在造模后12周和20周两个时间点对脑萎缩的程度进行对比分析,然后运用组织化学染色的方法观察在MRI上出现进行性萎缩的脑区中的神经元所发生的病理改变．MRI的结果表明：STZ诱导的T1DM大鼠相比于正常对照组大鼠出现了显著性的全脑体积、灰质体积和白质体积的萎缩,并且在多个白质脑区和灰质脑区均出现了萎缩程度随着病程的延长而逐渐加重．组织化学染色的结果发现,STZ诱导的T1DM大鼠相对于正常对照组大鼠在体感皮层、运动皮层和海马CA3区,均出现明显的神经元萎缩现象．     

组织蛋白酶B响应的超极化129Xe MRI探针对肺癌细胞的超灵敏探测

组织蛋白酶B（Cat B）是一种溶酶体半胱氨酸蛋白酶,在细胞代谢中起重要作用.已有研究表明Cat B在肺癌细胞中会过表达.因此,细胞内Cat B水平的检测非常重要.迄今为止,细胞内Cat B的检测方法主要为荧光成像,但该技术受限于渗透性和自发荧光背景干扰.为了解决这些问题,我们设计了一种基于超极化¹²⁹Xe磁共振成像的新型探针.它由一个作为¹²⁹Xe核磁共振（NMR）报告基团的穴番分子笼和一个作为Cat B特异性可裂解基团的酰胺键组成.当探针与Cat B相互作用时,酰胺键的断裂会导致其¹²⁹Xe化学位移发生变化.结合超极化-化学交换饱和转移（Hyper-CEST）技术,可为Cat B提供一种新颖的检测方法.     

一种示踪干细胞的磁共振-荧光双模成像探针

本文设计并合成了Gd基磁共振-荧光双模成像探针——Gd-DOTA-PEG-GA,通过电穿孔的方式标记人源间充质干细胞（hMSCs）.电穿孔标记诱导细胞将探针组装成团簇状纳米粒子进入细胞质,显著延长其与细胞结合的时间,并呈现出明显的T₂信号减弱效应,且信号减弱效应可以持续7天以上.在水溶液中,该探针的发射带集中在498 nm,并且荧光强度在一周内无明显衰减.该探针标记的细胞在荧光倒置显微镜下呈现绿色荧光.这些结果表明该探针可以作为磁共振-荧光双模成像探针用于干细胞示踪.     

基于动态有机钆纳米颗粒的T1-T2双模态MRI造影剂

本文设计、合成并测试了一种新型的基于有机钆纳米颗粒的磁共振成像（MRI）造影剂.以1, 2-氨基硫醇与氰基的缩合反应为基础,成功合成了粒径在8~23 nm范围内的有机钆纳米颗粒.该有机钆纳米颗粒作为磁共振造影剂时,随着时间的推移,其纵向弛豫率逐渐减弱,横向弛豫率先增强后逐渐减弱,这与钆纳米颗粒粒径增大有关.有机钆纳米颗粒同时存在随时间变化的纵向弛豫和横向弛豫,表明它有望成为一种先进的T₁-T₂双模态MRI造影剂.     

叶酸偶联磁性白蛋白纳米球的体内外靶向性评价

该文从体内外水平,探讨叶酸偶联磁性白蛋白纳米球对人鼻咽癌细胞KB的靶向效应. 体外实验中,将叶酸靶向、非叶酸靶向和叶酸抑制组与人鼻咽癌细胞KB共孵育24 h后,通过普鲁士蓝染色、7.0 Tesla Micro-MR仪检测叶酸受体介导的靶向效应;体内试验中,首先建立裸鼠荷人鼻咽癌细胞KB皮下移植瘤动物模型,再通过MRI检测及病理组织学检查,评价叶酸偶联磁性白蛋白纳米球的靶向性. 体外MRI成像显示叶酸靶向组与人鼻咽癌细胞KB共孵育后T₂WI信号强度降低,普鲁士蓝染色则显示靶向组细胞内存在大量铁颗粒;在体MRI成像结果显示,注射材料后不同时间点叶酸靶向组的T₂WI、T*₂-WI强度有明显下降,具有统计学差异. 病理学检查显示叶酸靶向组肿瘤组织内有较多蓝染的铁颗粒,而非靶向组和叶酸抑制组则未见铁颗粒的存在. 体内外实验均表明叶酸偶联磁性白蛋白纳米球对人鼻咽癌细胞KB具有良好的靶向性.     

常规T2像和组织学研究APPswe/PS1dE9转基因小鼠模型

用常规的T₂加权像研究阿尔茨海默病双转基因APPswe/PS1dE9小鼠模型,拟合了海马和顶叶皮层的T₂,并用组织学方法研究了APP/PS1小鼠在这2个感兴趣区域随年龄相关的淀粉样斑块病理的发展及铁质的沉积. T₂拟合的结果显示,较高月龄的APP/PS1小鼠相比于低月龄APP/PS1小鼠T₂有升高但不显著,各月龄组APP/PS1小鼠与同窝生非阳性小鼠之间也无明显差异;组织学的结果则显示年龄相关的淀粉样斑块的增多,铁质在16月龄的APP/PS1小鼠脑区中有明显沉积且和斑块具有“共域性”. 两方面的实验结果提示：在该模型淀粉样斑块沉积病理发展的早期,这2个脑区的T₂并不能作为特异性的病理发展的标志.     

超极化129Xe磁共振分子影像学

磁共振分子影像学发展的主要瓶颈之一在于灵敏度的限制,基于激光光泵和自旋交换技术能获得增强4~5个量级的超极化¹²⁹Xe磁共振信号,因此超极化¹²⁹Xe磁共振分子影像学相对于传统MRI在灵敏度上表现出巨大的优势. 围绕提高灵敏度这一核心MRI问题及其在科学研究中的应用,该文介绍了目前基于超极化¹²⁹Xe的生物分子探针的基本结构和原理,阐述了与之相关的分子影像学方法和技术,同时评述了当前的最新研究进展和发展方向.     

肿瘤细胞中表达的GFP蛋白的荧光漂白特性的研究

GFP作为生物源性荧光探针具有其他荧光标记物所无法比拟的优势，目前已广泛应用于生物学研究的各个领域。利用常规转染方法将带有EGFP基因的质粒载体导入人肺腺癌肿瘤细胞(ASTC-a-1)，并得到GFP稳定表达的细胞株。研究中发现，肿瘤细胞中表达的GFP长时间暴露于强激发光中会发生非常强列的荧光漂白作用，并且这种漂白作用是不可恢复的。对不同强度的激发光(饱和光源、阻断片ND4／ND8／ND16)对GFP的漂白作用进行了研究，并对冷冻保存样品的光漂白作用进行了初步的探讨。     

Dextran‐Coated Antiferromagnetic MnO Nanoparticles for a T1‐MRI Contrast Agent with High Colloidal Stability

A simple approach to synthesize carboxymethyl dextran‐coated MnO nanoparticles (CMDex‐MnONPs) with high colloidal stability in physiological saline solutions is described here for potential applications as a magnetic resonance imaging (MRI) T₁ contrast agent. The thermal decomposition methodology is used to produce uniform MnONPs with an average size of around 20 nm, and its hydrophobic surface is modified with CMDex molecules, conferring hydrophilic properties. After CMDex coating, the nanoparticle presents high colloidal stability in concentrations ranging from 10 to 50 μg mL^?1, average hydrodynamic size (Z‐average) of 130 nm, polydispersity degree of ≈12%, and negative surface charge in both simulated body fluid solutions and pure water with zeta‐potential of –20 and –40 mV, respectively. The CMDex‐MnONPs with 20 nm show antiferromagnetic behavior at room temperature, and the magnetic properties are found to be strongly dependent of the nanoparticle size, increasing the contribution of the ferromagnetic Mn₃O₄ phase with decreasing size for nanoparticles about 3 nm. Cytotoxicity evaluation in cancerous and noncancerous cells in the range of 5.0–50.0 μg mL^?1 shows low toxicity for cancerous cells and lack of the same for healthy cells lines. Related to the magnetic properties, CMDex‐MnONP presents significant r₁ relaxivity and low r₂/r₁ relaxivity ratio. The results suggest that these nanoparticles display characteristics for potential applications as an MRI T₁ contrast agent.     

Laser-Induced Fluorescence Bronchoscopy for Detection and Localization of Early Lung Cancer

Experimenatal results on the development of a Laser-Induced Fluorescence Bronchoscopy(LIFB) for the detection and localization of early lung cancer are reported in this paper. The system utilizes fluorescence of photosensitizer drug to provide real time video imaging for the examined lung tissue. Color filters are used to differentiate signal from background and a computer image processing technique is also applied to subtract the background. Moreover, a pseudocolor contrast enhancement method was developed to enhance the fluorescence image displayed on the vidio monitor. Suspicious areas are identified by pseudocolor image to guide biopsy, and several clinical trials show that sensitivity and contrast capability of the system should permit the detection and localization of early lung cancer.     

Low‐Magnetization Magnetic Microcapsules: A Synergistic Theranostic Platform for Remote Cancer Cells Therapy and Imaging

Multifunctional magnetic microcapsules (MMCs) for the combined cancer cells hyperthermia and chemotherapy in addition to MR imaging are successfully developed. A classical layer‐by‐layer technique of oppositely charged polyelectrolytes (poly(allylamine hydrochloride) (PAH) and poly(4‐styrene sulfonate sodium) (PSS)) is used as it affords great controllability over the preparation together with enhanced loading of the chemotherapeutic drug (doxorubicin, DOX) in the microcapsules. Superparamagnetic iron oxide (SPIOs) nanoparticles are layered in the system to afford MMC1 (one SPIOs layer) and MMC2 (two SPIOs layers). Most interestingly, MMC1 and MMC2 show efficient hyperthermia cell death and controlled DOX release although their magnetic saturation value falls below 2.5 emu g^?1, which is lower than the 7–22 emu g^?1 reported to be the minimum value needed for biomedical applications. Moreover, MMCs are pH responsive where a pH 5.5 (often reported for cancer cells) combined with hyperthermia increases DOX release predictably. Both systems prove viable when used as T2 contrast agents for MR imaging in HeLa cells with high biocompatibility. Thus, MMCs hold a great promise to be used commercially as a theranostic platform as they are controllably prepared, reproducibly enhanced, and serve as drug delivery, hyperthermia, and MRI contrast agents at the same time.     

A reference agent model for DCE MRI can be used to quantify the relative vascular permeability of two MRI contrast agents

Dynamic Contrast Enhancement (DCE) MRI has been used to measure the kinetic transport constant, K^trans, which is used to assess tumor angiogenesis and the effects of anti-angiogenic therapies. Standard DCE MRI methods must measure the pharmacokinetics of a contrast agent in the blood stream, known as the Arterial Input Function (AIF), which is then used as a reference for the pharmacokinetics of the agent in tumor tissue. However, the AIF is difficult to measure in pre-clinical tumor models and in patients. Moreover the AIF is dependent on the Fahraeus effect that causes a highly variable hematocrit (Hct) in tumor microvasculature, leading to erroneous estimates of K^trans. To overcome these problems, we have developed the Reference Agent Model (RAM) for DCE MRI analyses, which determines the relative K^trans of two contrast agents that are simultaneously co-injected and detected in the same tissue during a single DCE-MRI session. The RAM obviates the need to monitor the AIF because one contrast agent effectively serves as an internal reference in the tumor tissue for the other agent, and it also eliminates the systematic errors in the estimated K^trans caused by assuming an erroneous Hct. Simulations demonstrated that the RAM can accurately and precisely estimate the relative K^trans (R^Ktrans) of two agents. To experimentally evaluate the utility of RAM for analyzing DCE MRI results, we optimized a previously reported multiecho ¹⁹F MRI method to detect two perfluorinated contrast agents that were co-injected during a single in vivo study and selectively detected in the same tumor location. The results demonstrated that RAM determined R^Ktrans with excellent accuracy and precision.     

A 4-Methylumbelliferone-based Fluorescent Probe for the Sensitive Detection of Captopril

A highly sensitive fluorogenic probe for captopril, 4-methylumbelliferyl-2, 4-dinitrobenzenesulfonate (4-MUDNBS), was designed and synthesized. 4-MUDNBS is a nonfluorescent compound and was synthesized via the one-step reaction of 4-methylumbelliferone (4-MU) with 2,4-dinitrobenzenesulfonyl chloride. Upon mixing with captopril in basic solution, the 2,4-dinitrobenzenesulfonyl group of 4-MUDNBS was efficiently removed and highly fluorescent 4-MU was released, hence leading to the dramatic fluorescence increase of the reaction solution. The fluorescence intensity is linear with captopril concentration in the range 3.0–500 ng mL⁻¹ with a detection limit of 2.2 ng mL⁻¹ (3σ). The effect of substituents on the benzenesulfonyl moiety of the probe is discussed, and the presence of electronegative groups is favorable for the thiolate-induced cleavage reaction. The proposed method has been successfully applied to the captopril determination in pharmaceutical preparations.