靶向性荧光探针在乳腺癌早期诊断中的应用

选用E1/E3缺失型腺病毒作为靶向探针的载体,用化学修饰法将叶酸共价偶联腺病毒衣壳蛋白以提高探针的靶向性,用荧光染料罗丹明B偶联叶酸修饰的腺病毒,制备叶酸-腺病毒-RB探针,在体外研究此探针的毒性和靶向性.用近红外荧光染料ICG共价结合叶酸修饰的腺病毒,合成叶酸-腺病毒-ICG探针,使用近红外成像技术在位实时监测探针在体内的靶向性.实验结果证明叶酸-腺病毒-RB和叶酸-腺病毒-近红外探针对正细胞毒性较小,对早期乳腺癌细胞/组织具有较好的靶向性.     

H-22细胞声孔效应的应力阈值

理论和实验研究了超声空化场中的H-22型肝癌细胞产生可逆声孔效应的剪应力阈值.本文用1.37 MHz的聚焦声场,当超顺磁性纳米氧化铁在细胞悬液中的终浓度为410 μg/mL,换能器负载电功率为2 W,超声辐照60 s,细胞存活率90%以上时,有45.9±13.5%的细胞显示普鲁士蓝染阳性,暗示超声作用下,这些细胞表面曾出现可逆性微孔而使磁性微粒由此进入细胞内.利用无界自由空间微泡运动方程的球对称稳态解对实验条件下细胞膜表面的切变应力进行数值估算,结果表明,使H-22细胞产生可逆性声孔效应的微流剪应力阈值为697 Pa. 关键词： 声孔效应 磁性标记 微流 剪应力     

刀锋伪影校正技术在精神疾病患者海马MRI检查中的应用

为探讨磁共振刀锋伪影校正（BLADE）技术提升精神疾病患者海马磁共振图像质量的效果，本文分别使用结合了BLADE技术的BLADE T₂WI TSE、BLADE T₂WI FLAIR及传统T₂WI TSE、T₂WI FLAIR四种序列，对47例精神疾病患者和美国放射学院（ACR）标准模体在3.0 T磁共振成像（MRI）设备上分别进行常规海马斜冠状位扫描和ACR标准检测.患者的磁共振图像由2名放射科医师采用5分法对运动伪影、搏动伪影、颗粒度、海马磁共振图像质量进行评价，并应用Wilcoxon符号秩检验进行数据分析.模体图像通过识别图像的钻孔阵列和轮辐的数目，半定量评价各序列的高对比空间分辨力（HCSR）和低对比物体探测能力（LCD）.结果表明相比传统序列，结合BLADE技术的序列能够明显改善海马磁共振图像的运动伪影、搏动伪影（p<0.001），提高图像质量（p<0.05）；但在图像颗粒度方面，传统序列表现更优（p<0.001）.ACR模体半定量分析显示，结合BLADE技术序列与传统序列相比，在LCD检测方面结果更优、在HCSR检测方面结果相同或略逊.本文推荐将BLADE技术应用于不合作的精神疾病患者海马的MRI检查.     

多孔SiO2包裹磁性纳米颗粒Fe3O4的制备与表征

采用加热分解油酸铁法制备了Fe₃O₄磁性纳米颗粒,并用有机模板和反相微乳液相结合的方法将磁性纳米颗粒包裹在多孔二氧化硅中.用红外光谱(FTIR)研究了不同的处理方式对油酸铁表面官能团的影响及油酸的反应浓度和加热分解油酸铁的过程中升温速率对Fe₃O₄纳米颗粒的影响.结果表明,用乙醇和丙酮处理后的固态蜡状油酸铁表面的油酸基团会受到损害,将不利于加热分解时形成单分散性的Fe₃O₄纳 关键词： 3O₄纳米颗粒')" href="#">Fe₃O₄纳米颗粒 2包裹')" href="#">多孔SiO₂包裹 反相微乳液法 油酸铁     

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.     

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.     

靶向肺癌干细胞的多肽Gd基T1型MRI探针的研究

用固相多肽合成技术制备了特异性靶向肺癌干细胞的T_1型多肽核磁共振成像(MRI)探针——Gd-DOTA-HCBP-1.在11.7 T静磁场条件下,其纵向弛豫效率(r1)为6.15 mmol~(-1)·L·s~(-1),是商用T1造影剂Dotarem的1.6倍.体外细胞成像表明,此探针的使用可显著提高肺癌干细胞克隆球的可观测性,单个克隆球(包含2 000~4 000个细胞)可被明显辨识.     

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.     

MRI评价药物投递性能方法研究

采用MRI评价自制肝组织特异性非离子型高分子磁共振造影剂在小鼠体内药物投递效果. 分别在注射造影剂后0.1 h、6 h、12 h、24 h及7天采集磁共振T₁加权图像. 所有扫描均在1.5T临床磁共振成像仪上完成, 以固定体线圈为射频发射线圈, 三英寸圆形表面线圈为信号接收线圈. 数据分析前采用线圈非均匀性校正和信号非稳定性校正进行预处理. 实验结果显示,线圈空间敏感性校正使得小鼠组织图像信号强度空间更加均匀,稳定性校正后使得图像数据更加准确可靠,MRI是一种在体评价顺磁性标记高分子化合物药物投递效果的有效方法.     

SiO2纳米线簇、C-Si-O纳米球的制备及形貌、红外光谱和光致发光光谱研究

以二茂铁和硅油作为催化剂和原料,利用高温裂解硅油为C,Si,O源,在常压N₂和H₂混合气氛的化学气相沉积管式炉中制备了大量直径为5—40 nm、长数百纳米的非晶SiO₂纳米线簇及粒径为100—300 nm的C-Si-O实心纳米球. 利用透射电子显微镜、扫描电子显微镜对产物形貌进行表征.Fourier红外吸收谱显示出非晶SiO₂所具有的474,802和1100 cm^-1三个特征峰;SiO₂纳米线簇的光致发光光谱具有较强440 nm蓝光发光峰;而C-Si-O（原子数之比为1.13∶1∶2.35）纳米球具有奇特的红绿蓝（625,540,466 nm）三色光致发光谱. 关键词： 2纳米线簇')" href="#">SiO₂纳米线簇 C-Si-O纳米球 高温裂解 Fourier红外谱     

Magnetic resonance imaging of mouse Ehrlich carcinoma growth inhibition by thiacarpine, an analogue of cytotoxic marine alkaloid polycarpine

The anticancer effect of thiacarpine, a synthetic analogue of the known cytotoxic alkaloid polycarpine isolated from the Pacific ascidian Polycarpa aurata, was investigated in vivo in experiments using mouse solid Ehrlich carcinoma tumor as the target. A high-resolution magnetic resonance imaging (MRI) technique using a MR tomograph "PharmaScan" US70/16 (Bruker, Ettlingen, Germany) was used for visualization and quantification of tumor size. Fluorescence microscopy and image analysis were applied to determine Ehrlich carcinoma cell chromatin condensing (apoptosis) and necrosis in Ehrlich carcinoma cells at the action of thiacarpine in in vitro experiments. The scan and size calculations of the tumor and some mouse organs were carried out during the experiments. Thiacarpine in a total dose of 100 mg/kg was found to exhibit the delay in growth of the mouse tumor. The antineoplastic effect of this compound was accompanied by an increase in the lifetime of experimental mice in comparison with the control group of animals. Our data show that the ability of thiacarpine to induce apoptosis in carcinoma cells may contribute to thiacarpine anticancer effects against mice solid Ehrlich carcinoma in vivo detected by MRI.     

Red/Near‐Infrared Emissive Metalloporphyrin‐Based Nanodots for Magnetic Resonance Imaging‐Guided Photodynamic Therapy In Vivo

Near‐infrared emissive (NIR) porphyrin‐implanted carbon nanodots (PCNDs or MPCNDs) are prepared by selectively carbonization of free base or metal complexes [M = Zn(II) or Mn(III)] of tetra‐(meso‐aminophenyl)porphyrin in the presence of citric acid. The as‐prepared nanodots exhibit spontaneously NIR emission, small size, good aqueous dispersibility, and favorable biocompatibility characteristic of both porphyrins and pristine carbon nanodots. The subcellular localization experiment of nanodots indicates a lysosome‐targeting feature. And the in vitro photodynamic therapy (PDT) results on HeLa cells indicate the nanodots alone have no adverse effect on tumor cells, but display remarkable photodynamic efficacy upon irradiation. Moreover, MnPCNDs containing paramagnetic Mn(III) ions, which possesses good biocompatibility, NIR luminescence, and magnetic resonance imaging and efficient singlet oxygen production, are further studied in magnetic resonance imaging‐guided photodynamic therapy in vivo.     

Structural effect on degradability and in vivo contrast enhancement of polydisulfide Gd(III) complexes as biodegradable macromolecular MRI contrast agents

The structural effect of biodegradable macromolecular magnetic resonance imaging (MRI) contrast agents, polydisulfide gadolinium (Gd)(III) chelates, on their in vitro degradability, and cardiovascular and tumor imaging were evaluated in mice. Polydisulfide Gd(III) chelates, Gd-DTPA cystamine copolymers (GDCC), Gd-DTPA l-cystine copolymers (GDCP), Gd-DTPA d-cystine copolymers (dGDCP) and Gd-DTPA glutathione (oxidized) copolymers (GDGP), with different sizes and narrow molecular weight distribution were prepared and evaluated both in vitro and in vivo in mice bearing MDA-MB-231 tumor xenografts. GDGP with large steric hindrance around the disulfide bonds had greater T(1) and T(2) relaxivities than GDCC, GDCP and dGDCP. The degradability of the polydisulfide by the endogenous thiols decreased with increasing steric effects around the disulfide bonds in the order of GDCC>GDCP, dGDCP>GDGP. The size and degradability of the contrast agents had a significant impact on vascular contrast enhancement kinetics. The agents with a large size and low degradability resulted in more prolonged vascular enhancement than the agents with a small size and high degradability. It seems that the size and degradability of the agents did not significantly affect tumor enhancement. All agents resulted in significant contrast enhancement in tumor tissue. This study has demonstrated that the vascular enhancement kinetics of the polydisulfide MRI contrast agents can be controlled by their sizes and structures. The polydisulfide Gd(III) chelates are promising biodegradable macromolecular MRI contrast agents for magnetic resonance angiography and cancer imaging.     

Role of diffusion tensor imaging metrics and in vivo proton magnetic resonance spectroscopy in the differential diagnosis of cystic intracranial mass lesions

The purpose of this study was to determine whether proton magnetic resonance spectroscopy (PMRS) and diffusion tensor imaging (DTI) indices, fractional anisotropy (FA) and mean diffusivity (MD) can be used to distinguish brain abscess from cystic brain tumors, which are difficult to distinguish by conventional magnetic resonance imaging (MRI). Fifty-three patients with intracranial cystic mass lesions and 10 normal controls were studied. Conventional MRI, PMRS and DTI of all the patients were performed on a 1.5-T GE scanner. Forty patients were with brain abscess and 13 with cystic tumors. Cytosolic amino acids (AAs) were present in 32 of 40 brain abscess patients. Out of 13 patients with cystic tumors, lactate and choline were seen in 3 and only lactate was present in 10 patients on PMRS. All 40 cases of abscess had high FA, while all 13 cases of tumor cysts had high MD values. We conclude that FA measurements are more sensitive in predicting the abscess, while PMRS and MD are more specific in differentiating abscess from cystic tumors. We suggest that PMRS should be combined with DTI rather than with diffusion-weighted imaging as FA can be used as an additional parameter for separation of abscess from other cystic intracranial mass lesions.     

Positive contrast visualization for cellular magnetic resonance imaging using susceptibility-weighted echo-time encoding

Objective

The objective of this study was to investigate a method to generate positive contrast, selective to superparamagnetic iron oxide (SPIO) labeled cells, using the susceptibility-weighted echo-time encoding technique (SWEET).

Materials and Methods

SPIO-labeled human epidermal carcinoma (KB) cells were placed in a gel phantom. Positive contrast from the labeled cells was created by subtraction between conventional spin-echo images and echo-time shifted susceptibility-weighted images. SPIO-labeled cells were injected into the left dorsal flank and hind limb of nude mice, and unlabeled cells were placed on the right side as controls. Tumor growth was monitored using the proposed method, and a histological analysis was used to confirm the presence of the labeled cells.

Results

Based on in vitro testing, we could detect 5000 labeled cells at minimum and the number of pixels with positive contrast increased proportionally to the number of labeled cells. Animal experiments also revealed the presence of tumor growth from SPIO-loaded cells.

Conclusions

We demonstrated that the proposed method, based on the simple principle of echo-time shift, could be readily implemented in a clinical scanner to visualize the magnetic susceptibility effects of SPIO-loaded cells through a positive-contrast mechanism.     

In vitro and in vivo investigations of targeted chemotherapy with magnetic nanoparticles

Magnetic drug targeting is a local drug delivery system. Electromicroscopic pictures document the ferrofluid enrichment in the intracellular space in vitro. In vivo experiments were performed in VX2 tumor-bearing rabbits using magnetic nanoparticles bound to mitoxantrone. High-pressure liquid chromatography (HPLC) analyses after magnetic drug targeting showed an increasing concentration of the chemotherapeutic agent in the tumor region compared to regular systemic chemotherapy.     

The gadolinium complexes with polyoxometalates as potential MRI contrast agents

The two gadolinium (Gd) polyoxometalates, K(15)[Gd(BW(11)O(39))(2)] [Gd(BW(11))(2)] and K(17)[Gd(CuW(11)O(39))(2)] [Gd(CuW(11))(2)] have been evaluated by in vivo and in vitro experiments as the candidates of potential tissue-specific magnetic resonance imaging (MRI) contrast agents. T(1) relaxivities of 17.12 mM(-1) x s(-1) for Gd(BW(11))(2) and 19.95 mM(-1) x s(-1) for Gd(CuW(11))(2) (400 MHz, 25 degrees C) were much higher than that of the commercial MRI contrast agent (GdDTPA). Their relaxivities in bovine serum albumin and human serum transferrin solutions were also reported. After administration of Gd(BW(11))(2) and Gd(CuW(11))(2) to Wistar rats, MRI showed longer and remarkable enhancement in rat liver and favorable renal excretion capability. The signal intensity increased by 37.63+/-3.45% for the liver during the whole imaging period (100 min) and by 61.47+/-10.03% for kidney within 5-40 min after injection at 40+/-1-micromol x kg(-1) dose for Gd(CuW(11))(2), and Gd(BW(11))(2) induced 50.44+/-3.51% enhancement in the liver in 5-50-min range and 61.47+/-10.03% enhancement for kidney within 5-40 min after injection at 39+/-4 micromol x kg(-1) dose. In vitro and in vivo study showed that Gd(BW(11))(2) and Gd(CuW(11))(2) are favorable candidates as tissue-specific contrast agents for MRI.     

MRI contrast agent for molecular imaging of the HER2/neu receptor using targeted magnetic nanoparticles

In this study, Trastuzumab modified Magnetic Nanoparticles (TMNs) were prepared as a new contrast agent for detecting HER2 (Human epidermal growth factor receptor-2) expression tumors by magnetic resonance imaging (MRI). TMNs were prepared based on iron oxide nanoparticles core and Trastuzumab modified dextran coating. The TMNs core and hydrodynamic size were determined by transmission electron microscopy and dynamic light scattering. TMNs stability and cytotoxicity were investigated. The ability of TMNs for HER2 detection were evaluated in breast carcinoma cell lines (SKBr3 and MCF7 cells) and tumor-bearing mice by MRI and iron uptake determination. The particles core and hydrodynamic size were 9 ± 2.5 and 41 ± 15 nm (size range: 15?C87 nm), respectively. The molar antibody/nanoparticle ratio was 3.1?C3.5. TMNs were non-toxic to the cells below the 30 ??g (Fe)/mL concentration and good stable up to 8 weeks in PBS buffer. TMNs could detect HER2 oncogenes in the cells surface with imagable contrast by MRI. The invivo study in mice bearing tumors indicated that TMNs possessed a good diagnostic ability as HER2 specific contrast agent by MRI. TMNs were demonstrated to be able to selectively accumulate in the tumor cells, with a proper signal enhancement in MRI T2 images. So, the complex may be considered for further investigations as an MRI contrast agent for detection of HER2 expression tumors in human.     

Cellular uptake of folate-conjugated lipophilic superparamagnetic iron oxide nanoparticles

We prepared five folate-conjugated lipophilic superparamagnetic iron oxide nanoparticles (F₅-Liposuperparamagnetic iron oxide nanoparticles(SPIONs), 5.5 and 11 nm) and investigated their cellular uptake with KB cells, which is one of the representative folate-receptor over-expressing human epidermoid carcinoma cells, using MRI. The cellular uptake tests with the respective 5.5 and 11 nm F₅-LipoSPIONs at a fixed particle concentration showed appreciable amount of receptor-mediated uptakes and the specificity was higher in 5.5 nm SPIONs, due to its higher folic acid (FA) density, without inhibition. However, the numbers of the particles taken up under FA inhibition were similar, irrespective of their sizes.     

Synthesis,characterization, and in vitro biological evaluation of highly stable diversely functionalized superparamagnetic iron oxide nanoparticles

In this article, we report the design and synthesis of a series of well-dispersed superparamagnetic iron oxide nanoparticles (SPIONs) using chitosan as a surface modifying agent to develop a potential T ₂ contrast probe for magnetic resonance imaging (MRI). The amine, carboxyl, hydroxyl, and thiol functionalities were introduced on chitosan-coated magnetic probe via simple reactions with small reactive organic molecules to afford a series of biofunctionalized nanoparticles. Physico-chemical characterizations of these functionalized nanoparticles were performed by TEM, XRD, DLS, FTIR, and VSM. The colloidal stability of these functionalized iron oxide nanoparticles was investigated in presence of phosphate buffer saline, high salt concentrations and different cell media for 1 week. MRI analysis of human cervical carcinoma (HeLa) cell lines treated with nanoparticles elucidated that the amine-functionalized nanoparticles exhibited higher amount of signal darkening and lower T ₂ relaxation in comparison to the others. The cellular internalization efficacy of these functionalized SPIONs was also investigated with HeLa cancer cell line by magnetically activated cell sorting (MACS) and fluorescence microscopy and results established selectively higher internalization efficacy of amine-functionalized nanoparticles to cancer cells. These positive attributes demonstrated that these nanoconjugates can be used as a promising platform for further in vitro and in vivo biological evaluations.