外加磁场作用下磁性Fe3O4纳米粒子对肝癌细胞增殖和凋亡的影响

目的：观察磁性四氧化三铁（Fe3O4）纳米粒子对肝癌细胞的体外作用,并研究外加稳恒磁场（SMF）或交变磁场（EMF）对FeID4纳米粒子作用的影响。方法：光镜下观察CBRH-7919细胞对Fe3O4纳米粒子的吞噬作用;MTT法检测Fe304纳米粒子对大鼠肝癌细胞株CBRH-7919的毒性及外加磁场的影响;流式细胞术检测外加磁场作用下Fe3O4纳米粒子对细胞凋亡及线粒体膜电位的影响。结果：光镜下可见CBRH-7919细胞吞噬大量Fe3O4纳米粒子入胞浆,且交变磁场作用下细胞的吞噬量增加。30-100μg／mLFe3O4纳米粒子作用于CBRH-7919细胞未产生细胞毒性,稳恒磁场对其作用无影响,而交变磁场能增加Fe3O4纳米粒子的毒性,使细胞活性降低、凋亡率增加、线粒体膜电位降低。结论：交变磁场能增加CBRH-7919细胞对Fe3O4纳米粒子的吞噬并产生细胞毒性。     

磁性纳米粒子作用于肝癌细胞的生物学效应的研究进展

磁性纳米粒子,是一类智能型的纳米材料,因其特有的性质,被广泛应用于生物医学领域,在肝癌的治疗方面也有大量的实验性研究和成果。研究和探索磁性纳米粒子治疗肝癌的新方法和途径,有着很大的现实意义。本文就磁性纳米粒子作用于肝癌细胞的生物学效应的研究现状和进展进行总结整理,从三个方面进行了综述:磁性纳米粒子直接作用于肝癌细胞,探索磁性纳米粒子的生物相容性、在肝癌细胞的分布方式以及磁性纳米粒子本身对肝癌细胞的生物学效应的影响;磁性纳米粒子协同外加磁场(稳恒磁场、极低频交变磁场和高频交变磁场)作用于肝癌细胞;磁性纳米粒子外加修饰(磁性白蛋白纳米颗粒、纳米磁流体、磁性脂质体等),作为药物载体作用于肝癌细胞。     

纳米级Fe3O4磁性粒子与人肝癌细胞HepG-2及人正常肝细胞L02作用的生物学行为的实验研究

目的:对纳米级Fe3O4磁性粒子与人肝癌细胞HepG-2及人正常肝细胞L02作用的生物学行为进行实验研究。方法:通过化学沉淀法制备粒径为10nm左右的纳米级Fe3O4磁性粒子,观察其表征;将不同浓度纳米级Fe3O4粒子加入培养液分别与HepG-2混合培养检测凋亡坏死率;将相同浓度粒子分别与HepG-2和L02混合培养,对两者作用的差异进行动态观察比较。结果:纳米级Fe3O4磁性粒子能在肝癌细胞HepG-2细胞内稳定存在72小时以上,有良好的生物相容性;透射电镜观察到Fe3O4磁性粒子主要分布于细胞的溶酶体及吞噬泡内。共培养1小时后即有较多的纳米磁性粒子进入HepG-2内,而3小时后才见L02细胞内有少量的磁性粒子进入。结论:此实验结果为磁性纳米粒子与肿瘤细胞微观结构的作用提供了有意义的实验数据,并可能对应用磁性纳米粒子治疗恶性肿瘤提供有价值的依据。     

硒代硫酸钠诱导HL60细胞凋亡——还原态硒的细胞毒性(英文)

细胞毒性研究认为Cd2+的释放是硒化镉(CdSe)纳米粒子的细胞毒性机制之一,而Se2-阴离子在纳米粒子中的毒性机制未知。作者研究了硒代硫酸钠(selenosulfate(SSeO3)2-)对HL60细胞的细胞毒性作用,发现10μmol/L的硒代硫酸钠可以显著抑制细胞活力,诱导细胞凋亡,出现了染色质凝聚、DNA ladder和G0/G1凋亡亚峰。线粒体膜电位显著降低的同时,促凋亡蛋白Bax的免疫荧光增加。结果表明还原态的Se2-阴离子有显著的细胞毒性作用,可以诱导HL60细胞凋亡。同时也暗示Se2-阴离子的释放可能是含Se2-纳米粒子(比如硒化镉的量子点)细胞毒性的机制之一。     

合胞素通过促进线粒体去极化诱导小鼠淋巴瘤细胞EL4细胞凋亡

目的探讨人类内源性病毒W家族囊膜蛋白合胞素(syncytin)对白血病细胞凋亡的影响及其信号机制。方法对小鼠淋巴瘤细胞EL4细胞稳转表达合胞素,显微镜观察细胞形态变化,流式细胞术检测细胞凋亡、细胞周期及线粒体膜电位情况,用试剂盒检测PKC和Caspase-3活性。结果合胞素稳定过表达后,EL4细胞形态和PKC活性无明显改变,细胞凋亡明显增加,线粒体膜电位去极化增加,Caspase-3活性增强。结论合胞素可诱导小鼠淋巴瘤细胞EL4细胞线粒体途径细胞凋亡,在白血病的发生发展过程中发挥重要作用。     

基于L-半胱氨酸修饰的Fe3O4@TiO2复合纳米颗粒体外光动力疗法灭活HL60细胞的试验研究

本文采用共沉淀法制备了L-半胱氨酸(L-Cys)修饰的Fe3O4包裹TiO2(Fe3O4@TiO2/L-Cys)复合纳米粒子。通过透射电子显微镜(TEM),X射线衍射(XRD)和傅立叶变换红外光谱仪(FTIR)对复合纳米粒子进行了表征,并讨论了复合纳米粒子对HL60细胞体外光动力疗法(PDT)灭活的影响。并对其PDT灭活机制进行了初步探索。试验表明,Fe3O4@TiO2/L-Cys复合纳米粒子分散性高,生物相容性好,对细胞的暗毒性更低,并可以有效增强靶向性,提高PDT灭活效率,在410nm波长的光激发下,光照剂量为18J/cm^2的情况下,当TiO2与Fe3O4的比例为1∶3时,整体PDT效率最高。PDT灭活效率可达69.36%。     

弱稳恒磁场环境下基于BiFeO_3@TiO_2复合纳米颗粒体外光动力疗法灭活HL60细胞试验研究

本文通过溶胶-凝胶法制备了BiFeO_3@TiO_2复合纳米颗粒,利用透射电镜(TEM)、X射线衍射(XRD)、荧光发光光谱等对纳米颗粒进行表征。研究采用Cell Counting Kit-8(CCK-8)法分别检测了在暗室条件、光照条件以及不同强度的弱稳恒磁场作用下,用终值质量浓度为50μg/mL纳米颗粒处理HL60细胞活性,试验结果表明:在暗室条件下,药物质量浓度为50μg/mL,与HL60细胞共同孵育12 h后,BiFeO_3@TiO_2复合纳米颗粒随着TiO_2外壳厚度增加,细胞的暗室相对存活率从78%增加到85%;在光照条件下,有弱稳恒磁场作用的BiFeO_3与TiO_2质量比为1∶2的BiFeO_3@TiO_2复合纳米颗粒对HL60细胞的PDT灭活效率最高达到78%,弱稳恒磁场环境增强了对HL60细胞的PDT灭活效率,这为对弱稳恒磁场环境下的光动力疗法治疗白血病肿瘤细胞的临床应用提供了参考。     

黄芩甙对肝癌细胞BEL-7402形态学的影响

目的观察黄芩甙对肝癌细胞BEL-7402凋亡的影响,同时观察对肝癌细胞形态及超微结构、线粒体超微结构、线粒体膜电位和细胞内Ca^2＋的影响,探讨线粒体损伤在黄芩甙诱导肝癌细胞凋亡中的作用及可能的机制。方法应用细胞培养技术培养肝癌细胞BEL-7402,光镜、倒置显微镜、扫描电镜、透射电镜观察细胞形态及超微结构的变化尤其是线粒体的变化,应用流式细胞仪检测细胞凋亡百分率及线粒体膜电位、细胞内Ca^2＋的改变,免疫组化法检测细胞Bcl-2、Pax蛋白表达。结果黄芩甙诱导肝癌细胞BEL-7402凋亡呈剂量依赖关系,细胞形态、超微结构及线粒体超微结构出现明显改变,降低肝癌细胞线粒体膜电位,使细胞内Ca^2＋增加,细胞Pax表达增加,广泛分布于胞核和胞质中,Bcl-2表达减少。结论黄芩甙诱导肝癌细胞BEL-7402凋亡,线粒体损伤在黄芩甙诱导肝癌细胞凋亡中起重要作用,其机制可能为抑制肝癌细胞Bcl-2蛋白表达,促进Pax蛋白表达及细胞内Ca^2＋增加,激发线粒体膜通透性转运孔开放,线粒体跨膜电位降低,使肝癌细胞凋亡。     

葡聚糖磁性纳米颗粒外加磁场对人树突状细胞基因转染效率的研究

目的研究葡聚糖磁性纳米颗粒（the dextran coated magnetic iron oxide nanoparticles,DMN）在外加钕一铁一硼稀土固定磁场的作用下对人树突状细胞转染效率以及安全性的影响。方法先通过磁力计对DMN进行分析;再将修饰有多聚赖氨酸（Poly-L—Lysine,PLL）的DMN携带绿色荧光蛋白pEGFP—Cl质粒报告基因,在钕-铁-硼稀土周定强磁场的作用下,体外转染人树突状细胞,用荧光显微镜直接观察和流式细胞仪检测来评价外加磁场对DMN作为人树突状细胞转染载体效率的影响;在转染后采用MTT比色法测定在磁场干预下的DMN对人树突状细胞增殖和功能的影响以了解其细胞毒性。结果DMN的核心直径〈30nm,具有明硅的超顺磁性,比饱和磁化强度也明显高于相同Fe3O4含量的普通磁块;DMN作为基因载体在外加磁场作用下,转染12h即可将报告基因转染至人树突状细胞内并成功表达,在荧光显微镜下可观察到绿色荧光细胞,24h转染率可达到最高（约为27％）,转染效率较未加磁场组提高了2～4倍。而且转染后的人树突状细胞增殖活性及功能未因DMN外加磁场及其作用时间的长短而受到影响。结论超顺磁性的DMN在外加磁场作用下可以明显、安全、有效地提高对人树突状细胞的转染效率。     

三氧化二砷通过线粒体途径诱导人白血病HL-60细胞凋亡

目的:研究三氧化二砷(Arsenic trioxide,ATO)对人白血病HL-60细胞凋亡的影响,并以线粒体通路为靶点探讨其可能的机制。方法:采用1μg/m L、5μg/m L及10μg/m LATO处理HL-60细胞24小时后,采用流式细胞术检测细胞凋亡情况,通过细胞内MDA与GSH含量检测反映氧化应激水平,采用免疫印迹法检测凋亡相关分子表达,并通过免疫荧光染色检测细胞线粒体膜电位(mitochondrial membrane potential,MMP)水平。结果:5μg/m L及10μg/m L ATO可显著诱导人白血病HL-60细胞凋亡,并显著增加其氧化应激水平,增加促凋亡分子Bax和Caspase-3的表达,而抑制抗凋亡分子Bcl-2的表达,降低HL-60细胞线粒体膜电位的水平。结论:一定剂量的ATO可诱导人白血病HL-60细胞凋亡,而这一作用可能是通过诱导线粒体相关性凋亡信号通路激活实现。     

综述——纳米材料在组织工程中的应用：磁性纳米材料在组织工程研究中的应用

磁性纳米材料因其独特的理化性质在组织工程研究中被广泛地应用．本文主要从磁性纳米材料的表面化学活性、磁学性质以及生物应用磁性纳米材料的主要合成方法等几方面,综述了近年来利用磁性纳米材料设计组织工程支架材料的相关研究进展,包括纳米条件下的生长因子及相关基因的包裹和释放、机械力学刺激、干细胞追踪以及细胞图案化．     

多核磁共振19F-MRI在分子影像学中的应用

随着杂核氟、钠、磷等探针和成像技术的发展以及磁共振成像设备和序列的优化,多核磁共振迅速崛起,尤其是其在分子影像方面的研究与应用使包括心血管、肿瘤等众多疾病从传统的形态学影像诊断模式转向早期分子影像精准诊治模式。其中,19F-MRI多核磁共振分子成像近年来备受瞩目。虽然19F-MRI的成像敏感度是1H-MRI的82%,但人体只有牙齿中含有少量的氟,因此无背底噪声的干扰。19F-MRI应用氟类探针,19F自然丰度100%,且无放射性。本文简述了多核磁共振在分子影像学中的应用,并重点介绍19F-MRI分子影像及其应用探针在精准诊治方面的应用。     

多核19F磁共振成像研究进展

~(19)氟(~(19)F)的磁共振成像(MRI)研究可追溯到35年以上。在这段时间里,~1H磁共振成像的蓬勃发展使磁共振成为影像医学的支柱,但~(19)F磁共振成像研究的进展却较为缓慢。然而最近几年,~(19)F磁共振成像的研究受到了广泛的关注。在某种程度上,这是由于MR成像中软件与硬件的发展,更因为分子影像学的概念的提出与发展。本文将对~(19)F多核磁共振成像的应用,特别是使用全氟化碳纳米成像探针的19F多核磁共振成像进行综述。     

Magnetic properties of a cobalt(II) dimer of pseudo tetrahedral geometry [Co2{(CO)9Co3C---COO}5, C14H19N2H]

The crystalline ion pair [Co₂{OOC---CCo₃(CO)₉}₅, C₁₀H₆(N(CH₃)₂)₂H] (1) presents unusual magnetic properties. The X-band EPR spectrum of 1 at room temperature presents two unresolved bands at g=1.98 and 4.55. At a low temperature (20 K), the cluster of clusters 1 presents a complicated spectrum with an intense signal at 1700 G. The magnetic susceptibility of 1 was fit to a two spin S₁=S₂=3/2 Heisenberg model, with J=11.2 cm⁻¹ and a g value of 2.3. There is no field dependence of the magnetization, which suggests intramolecular coupling between the two tetrahedral centers of the cluster. Molecular orbital modeling indicates a sigma path of exchange between two topologically non-equivalent cobalt(II) centers.     

Magnetic relaxation in V15 cluster: Direct spin-phonon transitions

In this article we propose a model of spin-vibronic relaxation in K₆[V^IV₁₅As₆O₄₂(H₂O)]·8H₂O, the so called V₁₅ cluster exhibiting the unique layered structure. The work is motivated by the recent observation of the Rabi oscillation [1] in this system and aimed to elucidate the nature of the relaxation processes. The model assumes that the spin-phonon coupling arises as a result of modulation of the isotropic and antisymmetric (Dzyaloshinsky-Moriya) exchange interactions in the central triangular layer of vanadium ions by the acoustic lattice vibrations. Within the pseudo-angular momentum representation the selection rules for the direct (one-phonon) transitions between Zeeman levels are derived and a special role of the antisymmetric exchange is underlined. The relaxation times related to one-phonon transitions in different ranges of the field are estimated within the Debye model for the lattice vibrations.     

7Li Nuclear Magnetic Resonance Study for the Determination of Li+ Properties in Neuroblastoma SH-SY5Y Cells

Abstract: Lithium has been used clinically in the treatment of manic depression. However, its pharmacologic mode of action remains unclear. Characteristics of Li⁺ interactions in red blood cells (RBCs) have been identified. We investigated Li⁺ interactions on human neuroblastoma SH-SY5Y cells by developing a novel ⁷Li NMR method that provided a clear estimation of the intra- and extracellular amounts of Li⁺ in the presence of the shift reagent thulium-1,4,7,10-tetrazacyclododecane- N,N ', N ", N ‴-tetramethylene phosphonate (HTmDOTP⁴⁻). The first-order rate constants of Li⁺ influx and efflux for perfused, agarose-embedded SH-SY5Y cells in the presence of 3 m M HTmDOTP⁴⁻ were 0.055 ± 0.006 (n = 4) and −0.025 ± 0.006 min⁻¹ (n = 3), respectively. Significant increases in the rate constants of Li⁺ influx and efflux in the presence of 0.05 m M veratridine indicated the presence of Na⁺ channel-mediated Li⁺ transport in SH-SY5Y cells. ⁷Li NMR relaxation measurements showed that Li⁺ is immobilized more in human neuroblastoma SH-SY5Y cells than in human RBCs.     

Magnetic resonance study of 13C reductively methylated glycophorin and glycophorin glycopeptides

¹³C nuclear magnetic resonance (n.m.r.) spectral data for ¹³C reductively methylated N-terminal tryptic glycopeptides and for ¹³C reductively methylated N-terminal glyco-octapeptides derived from homozygous glycophorins A^M and A^N are presented. Their ¹³C chemical shift data are compared with the previously published ¹³C n.m.r. data for ¹³C reductively methylated homozygous glycophorins A^M and A^N in order to investigate the means of display of the MN blood determinants by these species. The pH dependence of the ¹³C resonances of $\text{N}{}^{\mathrm{\alpha }}\text{,N-[}{}^{13}\text{C}\text{]}\text{dimethyl}$ leucine of glyco-octapeptide A^N and of $\text{N}{}^{\mathrm{\alpha }}\text{,N-[}{}^{13}\text{C}\text{]}\text{dimethyl}$ serine of glyco-octapepti A^M indicated that only a slight structural perturbation occurs at the N-terminus when a large portion of the glycoprotein molecule is removed. However, one structural ‘state’ of ¹³C reductively methylated glycophorin A^M is lost when the glyco-octapeptide A^M is produced. The ¹³C resonance of $\text{N}{}^{\mathrm{\alpha }}\text{,N-[}{}^{13}\text{C}\text{]}\text{dimethyl}$ leucine of glycooctapeptide A^N titrated with a $\text{p}\text{K}{}_{\mathrm{<mtext>a</mtext>}}$ of 7.7 (Hill coefficient $\text{～ 1}$). The ¹³C resonance of $\text{N}{}^{\mathrm{\alpha }}\text{,N-[}{}^{13}\text{C}\text{]}\text{dimethyl}$ serine, on the other hand, exhibited an unusual pH dependence, indicating the existence of some possible steric constraints or hydrogen bonding in this molecule. In comparison to the data obtained for ¹³C-labelled glycooctapeptide A^M molecule, the pH dependence of the chemical shift of the ¹³C resonance of $\text{N}{}^{\mathrm{\alpha }}\text{,N-[}{}^{13}\text{C}\text{]}\text{dimethyl}$ serine of tripeptide tri-L-serine is also presented. Circular dichroism (c.d.) spectra indicated that the reductive methylation technique does not cause a large perturbation of the glycophorin A molecule.     

Conformations of P2 Protein of Peripheral Nerve Myelin by Nuclear Magnetic Resonance Spectroscopy

High-resolution 1H NMR spectra of P2 protein from bovine peripheral nerve myelin indicate that the protein contains a high degree of tertiary structure in aqueous solution. Denaturation of the protein in urea solutions is a multi-step process. Binding of lysophosphatidylcholine micelles to the protein causes a conformational change and a broadening of NMR peaks from side chains of aromatic amino acid and methionine residues, with much less effect on upfield methyl resonances.     

Measurement of Free Intracellular Calcium in the Brain by 19F-Nuclear Magnetic Resonance Spectroscopy

We report the first measurement of the free intracellular calcium level in an actively metabolising intact cerebral tissue preparation. To this end, we applied the recently developed 19F-nuclear magnetic resonance calcium chelator, 5,5'-F2-1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (5FBAPTA), in superfused cerebral cortical slices to give values for the intracellular Ca2+ concentration of 350 and 480 nM, at external calcium concentrations of 1.2 and 2.4 mM, respectively. Under both conditions, the intracellular Ca2+ concentration was increased by depolarisation using a high external K+ concentration. Interleaved 31P spectra showed that the presence of the 5FBAPTA had a deleterious effect on the metabolic state of the tissue with an external Ca2+ concentration of 1.2 mM, but normal viability was maintained using 2.4 mM.     

Effects of N-Methyl-d-Aspartate on [Ca2+]i and the Energy State in the Brain by 19F- and 31P-Nuclear Magnetic Resonance Spectroscopy

The effects of N-methyl-D-aspartate (NMDA) on the free intracellular Ca2+ concentration [( Ca2+]i) and the energy state in superfused cerebral cortical slices have been studied using 19F- and 31P-nuclear magnetic resonance spectroscopy. [Ca2+]i was measured using the calcium indicator 1,2-bis(2-amino-5-fluorophenoxy)ethane-N,N,N',N'-tetraacetic acid (5FBAPTA). NMDA (10 microM) in the absence of extracellular Mg2+ caused the expected rise in [Ca2+]i but produced an impairment of the energy state: the phosphocreatine (PCr) content was decreased by 42%, and the Pi/PCr ratio was increased by 55%. There was no detectable change in ATP or free intracellular Mg2+ concentration. Increasing the NMDA concentration in the superfusing medium to 100 or 400 microM caused no further increase in [Ca2+]i or further decrease in PCr content, but the Pi/PCr ratio continued to rise. The impairment of the energy state preceded the effect on [Ca2+]i, and these changes were irreversible on return to control conditions. Repeating the experiments in the presence of 1.2 mM extracellular Mg2+ resulted in similar changes in the energy state, with no change in [Ca2+]i. The possibilities that the effects were due to membrane depolarisation or to the presence of 5FBAPTA within the tissues were eliminated. The results suggest that low concentrations (10 microM) of NMDA produce an impaired energy state independent of the presence of extracellular Mg2+ and that the decreased energy state is not due to the changes in [Ca2+]i, which are seen only in the absence of extracellular Mg2+.