PC12细胞凋亡及其细胞膜表面超微结构的原子力显微镜形态学观察

本文采用原子力显微镜(AFM)对PC12细胞的凋亡及其膜表面超微结构进行了实验研究，研究表明AFM可直接观察PC12细胞的生长情况，凋亡细胞膜发生皱缩、凹陷，并形成微绒毛消失的凋亡小体等现象。AFM有望发展成为一种研究细胞凋亡的工具。     

人红细胞老化过程中膜表面唾液酸变化的免疫AFM研究

分离人外周血的轻龄和老龄红细胞,采用麦胚凝集素和胶体金对红细胞表面的唾液酸残基进行细胞化学标记,然后在AFM下成像并进行数据分析.结果显示,老龄红细胞膜表面胶体金标记的颗粒数量明显低于轻龄红细胞表面,两者之间有显著差异.推测红细胞老化过程中细胞膜表面唾液酸含量的变化可能与老化细胞的处理机制有关.免疫胶体金标记结合原子力显微镜可在纳米尺度对膜受体蛋白进行定位和定量研究,拓展了原子力显微镜在生命科学领域的应用.     

海马神经元的原子力显微成像

原子力显微镜(AFM)对完整的细胞成像并同时进行微细结构观察尚有一定困难。本实验改进了标本制备的过程．用原子力显微镜对戊二醛固定的海马神经元进行扫描，建立了方便、实用的完整海马神经元自完整胞体至超微结构的原子力显微镜成像技术，并用改进的方法获得了完整海马神经元及其超微结构的清晰的三维成像，并发现了一些其它显微技术所不能发现的微细结构。这些结构包括：①海马神经元胞体的亚细胞部分及这些亚细胞部分所具有的不同功能；②神经突触的完整形态；③损伤细胞膜表面出现的孔洞；④通过神经突触所形成的神经网络结构。     

壳聚糖载药纳米微球对人肝癌细胞的杀伤作用及AFM探测

本文以5-氟尿嘧啶（5-Fu）为抗肿瘤药物模型,壳聚糖为载体,制备平均粒径约为250nm的5-氟尿嘧啶/壳聚糖（5-Fu/Cs）载药纳米微球。采用MTT法评价载药纳米微球在体外对人肝癌细胞BEL7402的杀伤率,并采用原子力显微镜探测用药处理前后的肿瘤细胞表面超微结构的变化。结果表明,载药纳米微球对肿瘤细胞的杀伤率在量效和时效上明显比同剂量单独5-Fu作用的杀伤率高;AFM结果显示5-Fu与载药纳米微球对人肝癌细胞BEL7402作用后的细胞膜表面超微结构的变化存在较大差异,载药纳米微球在肿瘤细胞表面的吸附作用或内吞作用,导致载药纳米微球在细胞表面的释药机制不同于单独药物的扩散模型。     

海藻酸薄膜表面超微结构的原子力显微镜研究

本文采用原子力显微镜（ＡＦＭ）对海藻酸生物薄膜表面的超微结构进行了实验研究,选择出最佳观察方法。结果发现薄膜上分布不规则的微孔,其长短孔径之比大约为３：２,微孔的中心距为孔径的２倍。     

单个人体黑素瘤细胞的原子力显微图像及拉曼光谱的研究

利用原子力显微镜对人体单个黑素瘤细胞进行扫描,观察其表面形貌,得到了人体黑素瘤细胞在原子力显微镜扫描下的清晰成像,介绍了原子力显微镜在观察单个人体黑素瘤细胞表面超微结构方面的优势.通过其原子力图像,发现黑素瘤细胞形态各异,表面粗糙,有大小不一的突起.同时给出了单个黑素瘤细胞的拉曼光谱,并对谱峰进行指认,从分子水平对黑素瘤细胞的结构进行分析.将原子力图像及拉曼散射光谱技术应用于人体黑素瘤的研究,为人类黑素瘤的早期和快速诊断提供实验依据.     

原子力显微镜观察改性p（HEMA-MMA）对角膜基质细胞表面结构的影响

利用原子力显微镜分别对在I-型胶原和bFGF表面改性的p（HEMA-MMA）及未改性的p（HEMA-MMA）材料表面对于角膜基质细胞的膜表面结构进行了分析。将角膜基质细胞接种于改性前后的材料表面,用原子力显微镜对不同表面生长的角膜细胞的三维形态和超微结构等方面进行了研究。结果表明,改性后材料表面细胞宽度更大,高度更低,并且铺展较为完全,与正常条件下培养的细胞形态相似。膜超微结构参数分析显示在改性后材料表面生长的细胞膜微区的平均粗糙度（Ra）、均方粗糙度（Rq）、平均高度（MeanHt）、中值高度（MedianHt）明显较高于改性前材料表面的细胞。     

显微成像技术在氧化石墨烯与A549细胞骨架相互作用研究中的应用

石墨烯及其衍生物纳米材料因具有独特的物理化学性质,被广泛应用于生物医学领域,常被用作抗肿瘤药物的载体。本研究旨在探索氧化石墨烯对非小细胞肺癌细胞A549的生长和迁移的影响及机制。主要实验方法包括live/dead染色、Transwell实验、免疫印迹及原子力显微镜、激光共聚焦显微镜、透射电子显微镜、扫描电子显微镜等显微成像技术。结果显示GO能够穿透细胞膜切入细胞,导致细胞骨架肌动蛋白丝的形态结构发生显著改变;GO能够大量吸附细胞裂解物中F-actin蛋白。这些结果提示GO进入细胞后,可能通过吸附F-actin,破坏细胞骨架的结构,进而影响A549细胞的生长和转移。这些发现为未来以GO为载体的抗肿瘤药物设计提供了新的理论依据。     

AFM观察稳恒磁场对细胞表面精细结构的影响

利用原子力显微镜(AFM)观察稳恒磁场(SMF)处理后,悬浮生长细胞(K562人白血病细胞)、贴壁生长细胞(人结肠癌SW480细胞)、小鼠肝癌细胞Hepal-6和原代小鼠肝细胞表面精细结构的变化,以了解SMF杀伤肿瘤细胞的可能机制.观察结果显示:随曝磁时间延长,SMF可在肿瘤细胞表面造成不同程度损伤,主要表现为细胞膜上出现许多大小不一的凹陷,且凹陷数量和直径随着曝磁时间延长而增加.与MTT检测相比,AFM观察到的各类细胞表面损伤远早于细胞的生长抑制.实验观察显示,悬浮生长细胞比贴壁生长细胞对磁场处理更为敏感,小鼠肝癌细胞比肝细胞对磁处理更敏感.实验结果显示,AFM能够及早观察到细胞表面因SMF作用而产生的精细结构方面的变化.     

扫描探针显微学研究细胞的超微结构

利用近场光学显微镜(SNOM)和原子力显微镜(AFM)研究细胞的超微结构。SNOM对传统的光学分辨极限产生了革命性的突破,能够在纳米距离内探测光与样品物质之间的相互作用,在超高光学分辨率下对细胞成像,这种技术无侵入性和破坏性,能在细胞的自然状态下进行观测     

低强度超声对鼻咽癌细胞线粒体膜电位的影响

目的:观察低强度超声对鼻咽癌细胞线粒体膜电位的影响.方法:以低强度超声(频率1.7 MHz,剂量1.35W/cm2)辐照鼻咽癌细胞,采用Rhodamine123染色送CLSM下检测鼻咽癌细胞线粒体膜电位的变化.结果:超声波能够量著损伤鼻咽癌CNE2细胞线粒体,辐照后孵育4h,CLSM 下光吸收值与对照组相比显著降低,结...     

Modifying effects of recombinant tumor necrosis factor on the metastatic potential of tumor cells

Cells of the Lewis lung carcinoma (3LL), B16 melanoma and Ehrlich carcinoma pre-treated in vitro by a recombinant tumour necrosis factor (rTNF) were studied for their spontaneous and experimental metastatic spreading. The rTNF (1000 u/ml) was determined to stimulate a metastatic potential of the Lewis carcinoma cells after their treatment with the factor for 6 h and, vice versa, to inhibit it after 96 h prolonged treatment. The efficacy of the stimulating effect of rTNF on a metastatic spreading depends on the peculiarities of the studied cells.     

Poxvirus virions: their surface ultrastructure and interaction with the surface membrane of host cells

Virions of vaccinia and orf viruses were examined by ultrahigh-resolution scanning electron microscopy using a non-coating method. Intracellular mature particles of vaccinia virus appeared to be covered with a net and ultrastructurally their surface consists of many fine ridges and globules, while the surfaces of orf virus mature particles recovered from infected cells consist of spirally running protrusions. The ridge-like structures of vaccinia virus were presumed to correspond to surface tubules shown by negative staining of this virus, while the spiral protrusions of orf virus were presumed to correspond to spiral threads having a criss-cross appearance by the same staining. Using scanning electron microscopy in which the samples were prepared by the conventional method, we observed: (i) many virions, i.e. one or two hundreds, or occasionally more reaching about one thousand particles, of the IHD strain of vaccinia virus, (ii) many or a moderate number of virions, i.e. about one hundred or fewer particles, of the 58 strain of cowpox virus and (iii) rather few virions, i.e. several tens or fewer particles, of the Iwate strain of orf virus on the free surface of each cell infected with these viruses. It must be noted that the number of virions detected considerably differed in respective cells examined. Virus budding was frequently observed at the cell surface of monolayer cells infected with vaccinia virus but it was never detected with cowpox or orf virus, indicating a difference in the mechanism of virus release between vaccinia and the other two viruses. When whole cells infected with vaccinia virus were examined by a combination of high-voltage and scanning electron microscopies, virions on the cell surface and those inside the cells were clearly differentiated. All virions on the cell surface had an envelope, and some of the envelopes had a slack and/or one or more bulges.     

纳米金棒诱导A549细胞产生自噬的机制研究

纳米金棒(AuNRs)具有众多独特的属性,已广泛运用于生物医学领域,但其是否具有潜在的生物危害尚有争议.作者运用了激光扫描共聚焦显微镜技术、western blotting技术和其他分子生物学方法从细胞氧化应激的角度探讨了AuNRs诱导A549细胞产生自噬的分子机制.研究结果表明,4μg·mL-1的AuNRs处理6 h能够诱导A549细胞自噬标志蛋白LC3-Ⅱ表达增加,LC3蛋白从细胞核转移至细胞质并形成自噬小泡.进一步研究发现,AuNRs能够降低A549细胞线粒体膜电位、ATP含量、UCP2蛋白表达水平以及细胞抗氧化能力并导致活性氧蓄积,后者可能最终引起细胞产生自噬.而10 mmol·L-1抗氧化剂NAC能够逆转上述线粒体及细胞功能的改变,并抑制自噬的发生.这一研究为深入认识其生物危害及可能机制提供了有力的实验证据.     

Artificial Immunogenic Cell Death Lipid Nanoparticle Functions as a Therapeutic Vaccine for Cancer

Anticancer drug-mediated induction of immunogenic cell death (ICD) blocks metastasis or recurrence in cancer cells by promoting specific immune activity against cancer antigens. However, this strategy has failed to afford adequate treatment efficiency. Overcoming the failure of ICD-mediated cancer therapy, lipid nanoparticles (LNPs) containing cancer cell surface proteins are synthesized using sonication and extrusion without microfluidics. In addition, these LNPs are decorated with high-mobility group box 1 protein and calreticulin, indicators of ICD, and named artificial ICD LNPs (AiLNPs). Administration of AiLNPs effectively targets dendritic cells (DCs) and induces DC activation in mice. Moreover, treating CT-26 tumor-bearing mice with AiLNPs inhibits tumor growth by inducing CT-26 antigen-specific T-cell immunity. Furthermore, AiLNPs containing Lewis lung carcinoma (LLC1) membrane proteins can prevent metastatic LLC1 tumor growth in the lung via LLC1 antigen-specific T-cell activation. Finally, AiLNPs synthesized with human breast cancer membrane proteins activate DC-mediated antigen-specific T-cell immunity, effectively killing tumor cells. Therefore, AiLNPs are expected to be developed as a patient-specific cancer treatment to prevent cancer recurrence and metastasis.     

Selective adhesion of tumor cells to the endothelium of the target organ in metastasis

The fragments of cell plasma membranes of two lines of metastatic tumour of A/He mice (lung adenocarcinoma--LA-cells and hepatoma HA-1--HA-cells) were used to prepare "hybrid" vesicles (LA- and HA-liposomes). Incorporation of the fragments of LA-cells into the bilayer vesicle increased the binding of LA-liposomes with lung almost by an order of magnitude, their trapping by liver being noticeably decreased. HA-liposomes were retained in the liver in larger amounts than the model phospholipid liposomes. It is assumed that the specificity of metastatic spreading into an organ is associated with the interaction of the plasma membrane of the tumour cells with the apical membrane of the target organ endothelium.     

Evaluation of the distribution of lectin receptors on the surface of tumor and transformed cells using methods of variation statistics

Statistical procedures were used to estimate lectin receptor distribution on the surface of ascite lymphoma cells, neuroblastoma C-1300 cells and of transformed human T- and B-derived lymphoid cell lines. Relationships between the arithmetic means and mean square variances for sample populations from each cell and ferritin- or colloidal gold-lectin combination were used to define four types of topographical distributions: uniform-ordered, uniform-random, random and clustered.     

Mitochondrial breakage induced by the herbicide paraquat in cultured human lung cells.

Although the intracellular toxic sites of paraquat, a herbicide toxic to human bodies, have remained unclear for a long time, we recently demonstrated paraquat-induced mitochondrial injury in rat lung and liver in vivo. In the present study, cultured human lung cells (A549 adenocarcinoma cell line) which received 0.15 mM paraquat (equivalent to 40 mg/kg i.v.) showed selective mitochondrial breakage at 6-24 hr and died at 36-48 hr. These results suggest that mitochondria are the initial toxic site of paraquat in vitro as well as in vivo in contrast to the previously proposed microsome theory.     

Quantitative evaluation of the surface membrane receptors of leukemic cells to blood serum transcobalamin-II

Serum protein transcobalamin II (TC-II) is responsible for transport of cobalamins into mammalian cells. A method of quantitative estimation of plasma membrane receptors of hemopoietic cells to TC-II cobalamin complex is suggested. Analysis of mouse leukemia L1210 cells includes the saturation of radiolabelled ligand-receptor complex with papain. The number of receptors and 57CoCNCbl content in one cell is determined by differentiated radioactivity count of solubilized protein complexes and of cytoplasm.