磁流体靶向热疗对小鼠胰腺癌的作用

 为探讨磁流体靶向热疗对小鼠胰腺癌的体外和动物治疗作用,利用前期建株的小鼠胰腺腺泡细胞癌株(MPC-83)分别进行体外热疗和动物实验.对MPC-83 进行水浴热疗,分别调热疗温度为37、42、46、50℃,作用30 min,显微镜观察细胞形态变化,流式细胞仪检测凋亡和坏死细胞百分比.选择4 周龄雌性昆明种小鼠,建立MPC-83 胰腺癌皮下肿瘤模型,观察磁流体热疗(46℃和50℃)对荷瘤小鼠的作用及其病理学变化.流式细胞仪检测46℃和50℃热疗细胞凋亡和坏死百分比分别为46.13%、89.33%,与对照组比较,差异具有统计学意义(P<0.05).热疗后第14 天,46℃和50℃热疗组肿瘤生长率分别为-0.64±0.73和-0.72±0.79,与3 个对照组比较,肿瘤生长受到明显抑制(P<0.05).病理学检查示磁流体对照组,在注射磁流体24 h 可见散在的磁性纳米微粒在一定范围内分布于肿瘤细胞之间,部分肿瘤细胞和吞噬细胞吞噬了磁性纳米微粒.热疗14 d 肿瘤完全消失的小鼠皮下组织未见肿瘤细胞,可见皮下残存磁性纳米微粒,被吞噬细胞吞噬.各对照组小鼠瘤体生长旺盛,细胞核浓染分裂,可见病理性核分裂像.磁流体靶向热疗可以达到杀伤胰腺癌细胞的理想温度,能有效抑制MPC-83 胰腺癌生长,延长小鼠生存期.     

肿瘤主动靶向磁性纳米粒子研究现状及其在肿瘤热疗中的应用

 磁性纳米粒子已广泛应用于肿瘤的成像和治疗,但限制其临床应用的重大障碍是纳米粒子在肿瘤部位不能达到足够的浓度。主动靶向磁性纳米粒子是磁性纳米粒子表面偶联特定的靶向配体,靶向性结合特定的肿瘤细胞。靶向配体的选择是提高主动靶向性的关键。主动靶向性提高了磁性纳米粒子在肿瘤组织内的浓度,减少对正常组织的毒性,从而使其在肿瘤成像与治疗成为可能。磁感应热疗利用磁介质在外加交变磁场的作用下感应发热,是一种新型的具有前景的肿瘤治疗手段。依靠磁性纳米粒子主动靶向性,磁感应热疗将更好地实现细胞内热疗,提高肿瘤治疗的疗效。     

磁热耦合下不同注射方式对温度场的影响

磁流体热疗是具有生物相容性的磁性纳米流体注射到靶向肿瘤组织,通过施加交变磁场使注射区域升温达到杀死肿瘤组织目的。热疗期间的温度分布是磁热疗的关键因素,它直接影响了治疗效果。为分析不同注射方式对治疗效果的影响,本文采用有限元方法,建立磁热耦合模型,分析亥姆霍兹线圈产生磁场的均匀性,讨论了不同电流大小对磁场强度以及温度分布的影响。为了提升治疗效果,分析了不同注射方式对温度分布的影响,并以有效治疗体积这一参数较好的量化了治疗效果。结果表明：亥姆霍兹线圈作为磁场发生装置产生的磁场均匀性较高,不均匀度偏差在1%以内,通入线圈的电流每增加1A,磁场强度增加195.3 A/m;外加磁场条件相同,采用中心点注射时,组织内的温度最高为49.8℃,有效治疗体积最大为1 156.4 mm3,效果最好;调节外加磁场,使组织内最高温度保持48℃时,采用多点注射时有效治疗体积最大为1 530.7 mm3,治疗效果最好。因此,采用多点注射技术可以使治疗区域温度分布较为均匀,达到更好的治疗效果。     

Numerical modeling and simulation of temperature distribution uncertainty subject to ferromagnetic thermoseeds hyperthermia

In the process of ferromagnetic thermoseeds mediated magnetic induction hyperthermia, the tempera- ture distribution in the tumor region is a key factor to determine the therapy effect. During the preoperative treatment planning, discrepancies of the treatment parameters may lead to the temperature distribution uncertainty within the target area. Inaccurate prediction of temperature distribution may induce the treatment failure; therefore, it would be significant to investigate the uncertainty of tissue temperature prediction caused by the disturbance of calculation parameters. In this paper, 3D temperature field and necrosis zone of tissues in ferromagnetic thermoseeds hyperthermia are simulated by the finite volume algorithm, and effects of parameter uncertainties on the temperature distribution are revealed. Results show that selecting appropriate magnetic field parameters for treatment is a priority to guarantee therapeutic effect. Thermoseed properties and blood perfusion rate are the obvious disturbing terms for temperature distribution, while the metabolic heat of bio-tissues can often be ignored within limits. Our investigation is of importance for guiding reasonable and optimal preoperative treatment planning.     

Heating effect and biocompati bility of bacteri al magnetoso mes as potential materials used in magnetic fluid hyperthermia

Magnetic fluid hyperthermia (MFH) promises to be a viable alternative in the treatment of localized cancerous tumors.The treatment consists of introducing nanoparticles as energy absorbent agents in tu...     

DSA 联合 CT 引导下经皮肝穿刺兔 VX2 肝癌模型制作

摘要: 目的 探讨 DSA 联合 CT 引导下经皮肝穿刺兔 VX2 肝癌模型制作的可行性。方法 40 只新西兰白兔随机分 成 A、B 两组,每组 20 只,A 组在 DSA 联合 CT 引导下将兔 VX2 肿瘤组织块经皮肝穿刺接种于兔左肝内,B 组通过 开腹法种植 VX2 肿瘤组织块,于接种后第 2 周、4 周行全身 MR 检查,后处死实验兔行原位、异位肿瘤肉眼观察。结 果 A 组建模成功率 90% ,有 1 只出现异位种植,未发现腹水及死亡,平均手术时间( 17. 3 ± 5. 2) min; B 组建模成 功率 85% ,有 3 只出现腹水,2 只死亡,无异位种植发生,平均手术时间( 50. 7 ± 11. 3) min,,两组手术时间比较差异 有统计学意义( P ＜ 0. 05) 。结论 DSA 联合 CT 引导下经皮肝穿刺制作兔 VX2 肝癌模型方法简单、建模成功率高、 手术操作时间短。     

p53基因联合热放疗治疗晚期软组织肉瘤的临床研究

 为评价重组人p53腺病毒注射液(rAdp53)结合热疗治疗软组织肉瘤的疗效及安全性,2001年10月至2012年6月,采用rAdp53,结合热疗放疗,治疗晚期软组织肉瘤30例.基因治疗采用瘤内注射或腹腔灌注rAdp53,每周1次,每次1×10¹²VP,平均8次.所有患者均配合热疗,平均热疗8次.放疗每次2Gy,每周5次,剂量为16~70Gy/8~35次/2~8周,平均56.3Gy.30例患者在治疗前后观测肿瘤变化,以CT评价治疗后2个月的疗效.结果表明,病灶缩小超过或等于50%者9例,介于25%~50%者8例,缩小小于25%者12例;病灶增大者1例.生存率1年为(58.6±0.091)%,2年为(22.4±0.079)%,3年为(11.2±0.069)%,4年为(5.6±0.052)%.30例患者共接受216次rAdp53瘤内注射,除1例出现过性发热外,未发现其他不良反应,未影响热疗和放疗.研究表明,软组织肉瘤瘤内注射rAdp53结合热疗、放疗安全、有效.rAdp53是一种很有潜力的治疗恶性软组织肿瘤的基因治疗药物.     

In vivo assessment of hepatotoxicity,nephrotoxicity and biodistribution using 3-aminopropyltriethoxysilane-coated magnetic nanoparticles （APTS-MNPs） in ICR mice

The biocompatibility and biodistribution of magnetic nanoparticles （MNPs） in vivo are essential to ensure their safely clinical application. We have studied these aspects with our 3-aminopropyltriethoxysilane-coated magnetic nanoparticles （APTS-MNPs） formulation, which can be used as magnetic induction hyperthermia media. Changes in tissue iron levels were analyzed after intraperitoneal injection of APTS-MNPs to ICR mice. Liver and kidney functions were tested. Heart, liver, spleen, lung, kidney, testis, and brain were sectioned for pathological analysis. Biodistribution of iron in various body tissues changed with time but greater fraction of the injected iron localized in the liver and spleen than in other tissues. Serum showed an increase in AST and LDH fol-lowing APTS-MNPs injection. Histological analyses of selected tissues showed no obvious abnormal changes. In conclusion, APTS-MNPs did not cause continuing changes in the liver and kidney function and thus can be safely used for in vivo application.     

Growth potential of human hepatocarcinoma cells in the liver of neonatal immunocompetent mice and its relation to immunological tolerance

To determine the pathological behavior of human hepatocarcinoma cells in the liver microenvironment of neonatal non-immunode-ficient mice, three human hepatocarcinoma cell lines (Bel7402, HepG2, and SK-Hep-1), traced by DiI, were transplanted into the intrahepatic or subcutaneous tissue of neonatal and adult Kunming mice. Histopathological observations showed that cells in the adult liver induced a severe immune response as early as the second day after the implantation, while the subcutaneous neoplasm underwent extensive necrosis by the end of the study. Only the cells injected into the neonatal liver underwent a delayed immunologic rejection in the organ microenvironment. These cells retained recognizable tumor features over the first seven days, and displayed an intrahepatic invasive pattern. The expression of tumor markers including alpha-fetoprotein and survivin was maintained. The quantitative ELISA for the expression patterns of IL-2 and IL-10 also confirmed that the intrahepatic immunity was non-susceptive during this period. The high serum alpha-fetoprotein level was inversely correlated with the change in immune response. Our study provided a bio-system for the research of immune responses to xenografts in the liver.     

磁流体热疗设备的三维电磁场模拟

为清楚了解磁流体热疗设备产生的电磁场的分布,本文采用有限元方法对自制的设备原型建立三维模型并进行了优化,计算了该模型在静态和谐态下产生的电磁场的分布,对磁极中间的气隙宽度在20～40mm范围内变化时的磁场强度进行了对比．选取气隙宽度为30mm时计算电磁场的磁场强度、电感和涡流损耗,所得结果与实验测量值完全吻合,验证了模拟计算的可靠性．     

用于高效磁转染的鱼精蛋白修饰的铁氧磁性纳米粒研究

 功能化铁氧磁性纳米粒在生物医学中应用广泛,可用于肿瘤磁感应热疗、磁共振成像（Magnetic Resonance Imaging,MRI）、药物输送及磁转染等方面。为了探讨鱼精蛋白功能化修饰的铁氧磁性纳米粒的制备及其作为基因载体在体外磁转染中的可行性,采用共沉淀法制备Fe3O4磁性纳米粒,经表面氨基化修饰后在其表面偶联鱼精蛋白。利用透射电镜、傅里叶红外光谱仪、zeta电位与粒度分析仪等,对磁性纳米粒进行形态、粒径及zeta电位分析等表征检测。共聚焦显微镜观察磁转染方法转染报告基因绿色荧光蛋白质粒pEGFP-N1进入HepG2细胞的表达,以真核转染试剂vigofect为对照。结果显示,实验中制备的磁性纳米粒粒径10nm左右,在交变磁场下具有良好的升温性能。鱼精蛋白功能化修饰磁性纳米粒后,其zeta电位进一步增大,更利于与DNA有效结合,在HepG2细胞系,其转染pEGFP-N1质粒的效率高于vigofect。研究表明,鱼精蛋白功能化修饰的铁氧磁性纳米粒可作为磁转染的有效载体,由于其同时具备在交变磁场下升温的性能,在基因治疗联合热疗的研究领域具有一定的应用价值。     

四氧化三铁纳米粒子与癌细胞相互作用的初步研究

用共沉淀法制备了粒径为7．5nm的Fe3O4纳米粒子，并用透射电镜(TEM)观察其形貌为分布均匀的球形．将Fe3O4纳米粒子加入癌细胞株7901和MKN-45的培养液中，培养一段时间后，通过TEM观察细胞的形貌，发现Fe3O4纳米粒子被摄入到癌细胞内，通过原子吸收光谱(AAS)测量了细胞对Fe3O4纳米粒子的摄入量．结果表明，Fe3O4纳米粒子可以逐渐被癌细胞摄入，在癌细胞内达到一定的浓度范围．该研究为利用Fe3O4纳米粒子的磁过热疗法治疗肿瘤提供了细胞层次的实验和理论基础．     

超声热疗的现状与应用

 超声热疗是利用生物组织吸收超声波能量使自身温度升高的特性,通过高温致死病变组织或低温促使病变组织康复的热疗技术.该热疗具有微创、安全、高效等特点.利用高温致死病变组织的高强度聚焦超声已应用于子宫肌瘤、乳腺肿瘤、前列腺肿瘤、肝肿瘤等的临床治疗,在脑肿瘤及神经系统疾病方面尚处于临床应用研究阶段.低强度超声的低温升热疗对于产后出血治疗和神经组织愈合等有较好疗效.本文综述高强度聚焦超声和低强度超声热疗技术的发展、现状及应用.     

Dialdehyde starch nanoparticles as antitumor drug delivery system: An in vitro, in vivo, and immunohistological evaluation

Sustaining the release of therapeutic nanoparticles in a cell-, tissue-, or disease-specific manner is a potentially powerful technology. A new drug carrier-dialdehyde starch nanoparticle (DASNP) that can sustain the loading and release of 5-fluorouracil (5-Fu) antitumor drug is reported in this study. IR spectrophotometer and 1H NMR confirmed the formation of aldehyde groups, and scan electron microscope determinations showed that the dialdehyde starch nanoparticles obtained had an average diameter of 90 nm. 5-Fu, the model drug, was conjugated into nanoparticles by aldehyde groups. These 5-Fu-binding nanoparticles significantly enhanced breast cancer cell (MCF-7) inhibition in vitro compared with free 5-Fu. After subcutaneous 0 injection in the breast tumor-loaded rats, 5-Fu-DASNP exhibited remarkable tumor-inhibitory efficacy determined by measuring tumor weight in vivo. The tumor inhibition of 5-Fu-DASNP was 61%±6%, whereas that of free 5-Fu was only 42%±4%. Bcl-2/Bax immunohistochem-istry studies indicated that 5-Fu-DASNP remarkably induced tumor tissue necrosis. These results demonstrated that the DASNP prepared in this work is a potentially effective drug carrier.     

磁感应治疗研究和临床试验

 肿瘤磁感应加温治疗是近年来兴起的一项新型肿瘤治疗技术。该技术利用定位导入肿瘤靶区的磁性介质在交变磁场下的磁化损耗产热,对恶性肿瘤实施磁介导加温治疗。研究表明,肿瘤组织在交变磁场诱导的磁热效应下升温至40～70℃时,靶区沉积的热能作用可特异性杀死癌细胞,抑制癌症复发和转移,并可激发机体的抗癌免疫效应,进一步增强治疗效果,是一种简便易行、安全有效的肿瘤治疗方法。磁感应加温治疗在肿瘤治疗的特异性、靶向性等方面显现出其独特优势,已引起了国内外肿瘤临床治疗界的极大兴趣。本文对实验室近年来开展的基于多种磁性介质的肿瘤磁感应热疗研究进行了综述,主要包括磁感应治疗设备研制、热疗用磁性介质研发和磁热疗生物医学实验研究3个方面。在这些研究的基础上,肿瘤磁感应热疗的临床前实验已经完成,目前已经开展一期临床试验研究。     

Amphiphilic dextran/magnetite nanocomposites as magnetic resonance imaging probes

Superparamagnetic iron oxide （SPIO） nanoparticles are effective contrast agents for enhancement of magnetic resonance imaging at the tissue, cellular or even molecular levels. High quality SPIO nanoparticles can be synthesized in the organic phase but need to be transferred into water before any biomedical applications. In this study, amphiphilic poly（ε-caprolactone） grafted dextran （Dex-g-PCL） was used as carriers for particle encapsulation and stabilization in the aqueous phase. Multiple SPIO nanoparticles were self-assembled together with the help of Dex-g-PCL during phase transfer from chloroform to water, and diameters of Dex-g-PCL/SPIO nanocomposites were （64 ± 22） nm through dynamic light scattering measurement. These nanocomposites were superparamagnetic at 300 K with saturated magnetization of 88 emu/g Fe. In the magnetic field of 1.5 T, Dex-g-PCL/SPIO nanocomposites had a T2 relaxivity of 363 Fe mL·mol-1·s-1. This unique nanocomposite brought significant mouse liver contrast with signal intensity changes of -60% at 5 min after intravenous administration. However, uptake of Dex-g-PCL/SPIO nanocomposites in liver reticuloendothelial cells （Kupffer cells） did not immediately happen at shorter time points （〈4 h） as verified by histology studies, and it was evident that more iron staining would be located in Kupffer cells 24 h after contrast agent administration. After 24 h and 10 d, the signal intensities （SI） gradually recovered, and SI changes were -44% and -31%, respectively. From our observation, the time window for enhanced-MRI could last at least 12 days and totally recovered after 16 days. This novel sensitive MRI contrast agent may find potential applications in discovering small liver lesions such as early tumor diagnosis.     

羧基磁流体制备和牛血清白蛋白表面修饰

 磁性纳米粒已被用作肿瘤磁感应热疗的介质，目前常用纳米Fe3O4，其经典制备方法是化学共沉淀法，得到具有超顺磁性的纳米Fe3O4粒径约为10～15nm。本文采用改进的化学共沉淀法制备聚丙烯酸（PAA）修饰的羧基纳米Fe3O4，通过EDC、NHS活化法偶联模型蛋白牛血清白蛋白（BSA），用BCA定量分析磁粒对BSA的偶联效率，并进一步偶联荧光标记的抗体，观察偶联效果。结果表明，改进的化学共沉淀法制备的羧基纳米粒粒径约10nm，水相分散性良好；定量分析和光学观察结果均显示，采用EDC、NHS活化法能使蛋白质与羧基化磁性颗粒稳定结合。本研究为靶向磁性纳米粒的制备提供一种新方法。     

Assessment of alternated cooling and heating treatment by US combined CEUS in the VX2 rabbit liver tumor model

Inoperable liver tumors are often treated by thermal ablation that destroys the tumor in situ and spares the adjacent hepatic tissue.Thermal–physical treatment has many advantages,but treatment by freezing or heating alone has some limitations.By taking the advantages and disadvantages of cryosurgery and thermotherapy into consideration,a new thermal technique that combines cryosurgery and radio frequency ablation has been proposed,thereby overcoming the disadvantages of each treatment strategy and improving therapeutic outcomes.This new approach remains to be systematically studied in the liver;therefore,this study was performed to estimate survival after alternated cooling and heating ablation therapy in a VX2 rabbit liver tumor model.Sixteen days after VX2 carcinoma implantation into the rabbit liver,tumors were treated with alternated cooling and heating ablation therapy.Rabbits were monitored for 6 months after treatment and assessed with ultrasound(US)and computed tomography at 1,7,14,and 30 days posttreatment.Untreated tumor-bearing animals served as the control group.Our results show that alternate freezing and heating ablation therapy resulted in a good recovery of VX2 rabbits.Compared with the control group,treated rabbits lived significantly longer(P\0.05),with 70%of treated animals surviving to 196 days posttreatment without metastasis or recurrence,while none of the controls did so.There was no local recurrence in the treatment group.All rabbits in the control group developed metastasis,while metastasis was only observed in 30% of treated rabbits.These results suggest that alternate cooling and heating ablation therapy can prolong the survival time of rabbits with VX2 liver tumors and is an effective method for tumor therapy.Furthermore,we also showed in this model that contrast enhanced US is a valid follow-up approach to assess treatment effectiveness.     

宫颈癌TIL在体内器官及肿瘤组织中的分布特征

研究了＾１２５Ｉ－ｕｄＲ标记的宫颈癌ＴＩＬ在荷瘤Ｃ５７／ＢＬ６小鼠体内的密度分布特征。结果表明，ＴＩＬ在血，肝，肺和肾脏等器官１次分布，再在肿瘤组织和脾脏２次分布；ＴＩＬ在外周血的分布速率最快，其次是肝，肺，脾，肾和肿瘤组织；ＴＩＬ对肿瘤组织的亲和性虽较脾，肺，肾为低，但仍比肝脏高。     

磁感应热疗联合125I籽源近距离放疗研究

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