Telomerase Activity and Microgravity-Dependent Apoptosis in Human Leukemia HL-60 Cells

Mammalian cells subjected to conditions of spaceflight and the microgravity environment ofspace; manifest a number of alterations in structure and function. Among the most notable changes incells flown on the Space Shuttle are reduced growth activation and decline in growth rate in the totalpopulation. Other changes include chromosomal aberrations, inhibited locomotion, alteredcytokine production, changes in PKC distribution, and increased apoptos.

大蹼铃蟾蛋白提取物对膀胱癌细胞株的抑制作用及对端粒酶的影响

目的探讨大蹼铃蟾蛋白提取物对膀胱癌细胞株的 抑制作用及对端粒酶的影响。方法采用肿瘤细胞培养方 法，检测大蹼铃蟾蛋白提取物对膀胱癌细胞株."%、/01’)+、 234/56的抑制作用；以.&-7’78&’9:02-方法检测大蹼铃 蟾蛋白提取物对膀胱癌细胞株端粒酶表达的影响。结果 大蹼铃蟾蛋白提取物表现出对膀胱癌细胞株有较强的抑制 作用；对膀胱癌细胞株端粒酶表达则无影响。结论大蹼铃 蟾蛋白提取物对多个膀胱癌细胞株具有直接细胞杀伤作用， 其作用机制不通过抑制端粒酶活性。

Staphylococcus aureus lipoproteins trigger human corneal epithelial innate response through toll-like receptor-2

Bacterial lipoproteins (LP) are a family of cell wall components found in a wide variety of bacteria. In this study, we characterized the response of HUCL, a telomerase-immortalized human corneal epithelial cell (HCEC) line, to LP isolated from Staphylococcus (S) aureus. S. aureus LP (saLP) prepared by Triton X-114 extraction stimulated the activation of NF-kappa B, JNK, and P38 signaling pathways in HUCL cells. The extracts failed to stimulate NF-kappa B activation in HUCL cells after lipoprotein lipase treatment and in cell lines expressing TLR4 or TLR9, but not TLR2, indicating lipoprotein nature of the extracts. saLP induced the up-regulation of a variety of inflammatory cytokines and chemokines (IL-6, IL-8, ICAM-1). antimicrobial molecules (hBD-2, LL-37, and iNOS), and homeostasis genes (Mn-SOD) at both the mRNA level and protein level. Similar inflammatory response to saLP was also observed in primarily cultured HCECs using the production of IL-6 as readout. Moreover, TLR2 neutralizing antibody blocked the saLP-induced secretion of IL-6, IL-8 and hBD2 in HUCL cells. Our findings suggest that saLP activates TLR2 and triggers innate immune response in the cornea to S. aureus infection via production of proinflammatory cytokines and defense molecules. (C) 2007 Elsevier Ltd. All rights reserved.

12C6+重离子辐照对细胞端粒酶活性表达的影响

端粒是染色体末端由重复DNA序列和相关蛋白组成的一种特殊结构，具有稳定染色体结构及完整性的功能，会随染色体复制与细胞分裂而缩短。端粒酶是一种核糖核蛋白，能以自身RNA模板合成端粒DNA，催化合成TTAGGG重复序列，添加到染色体末端，维持端粒长度不变。端粒酶主要由hTR, TEPl和hTERT组成，一般认为hTERT是端粒酶激活的限速因素。大多数永生化细胞和恶性肿瘤细胞具有端粒酶活性，因而端粒酶目前已成为为细胞持续分裂提供遗传基础的原因。由于端粒酶与细胞衰老、肿瘤发生等关系如此密切，因此已成为肿瘤放射治疗研究热点。目的： 本文利用兰州近代物理研究所重离子研究装置（Heavy Ion Research Facility in Lanzhou,HIRFL）产生的碳离子（31MeV/u 12C6+）对人体细胞、癌细胞进行不同剂量的辐照，探索细胞中端粒酶活性的变化及与之相关的生物学信息。材料与方法： 以人肝细胞HL－7702，肝癌细胞SMMC－7721为实验对象，用不同剂量1Gy、2Gy、3Gy、4Gy的重离子分别对两种细胞进行照射，用多聚酶链式反应-银染端粒重复序列扩增法（PCR- telomeric repeat amplification protocol，TRAP-PCR）检测不同剂量下细胞端粒酶活性的变化。并提取不同剂量下的细胞转入培养皿中，培养10天，固定，染色。统计大于50个细胞的克隆数，绘制细胞存活曲线。结果与讨论： 结果显示，人肝细胞HL-7702自身没有端粒酶活性，经1Gy辐照后也没有端粒酶活性，在2、3Gy处出现端粒酶活性，4Gy处端粒酶活性又消失。肝癌细胞SMMC-7721在1－3Gy处随着剂量的增大端粒酶活性升高，在4Gy处又开始下降；在1－3Gy处随着时间的推移端粒酶活性随着时间而加强（p<0.05）。随着剂量的增大，细胞存活率呈剂量依赖型下降。分析得知，重离子辐射可以诱导人肝细胞产生端粒酶活性，也可以改变肝癌细胞的端粒酶活性。端粒酶参与细胞受辐照后DNA单链损伤的修复；辐照后DNA双链断裂导致端粒酶活性减弱。本实验与其他低LET射线相比，使重离子在辐照治疗中的优势得以体现

G-quadruplex-based DNAzyme for facile colorimetric detection of thrombin

Thrombin-binding aptamer is found to bind hemin to form a catalytic complex whose activity is significantly promoted by the addition of thrombin, which enables the colorimetric detection of thrombin with high specificity and sensitivity in a facile way.

Chemiluminescence thrombin aptasensor using high-activity DNAzyme as catalytic label

With a designed high-activity DNAzyme as the catalytic label, an ultrasensitive chemiluminescence thrombin aptasensor is developed, enabling a 10- to 100-fold improvement in the detection sensitivity as compared with previous counterparts.

Polyethyleneimine-Functionalized Platinum Nanoparticles with High Electrochemiluminescence Activity and Their Applications to Amplified Analysis of Biomolecules

Polyethyleneimine-functionalized platinum nanoparticles (PtNPs) with excellent electrochemiluminescence (ECL) properties were synthesized and applied to the amplified analysis of biomolecules. These particles were prepared at room temperature, with hyperbranched polyethyleneimine (HBPEI) as the stabilizer. The UV/Vis absorption spectra and transmission electron microscopy images clearly confirmed the formation of monodisperse PtNPs. Such particles proved to possess high stability against salt-induced aggregation, enabling them to be employed even under high-salt conditions. Owing to the existence of many tertiary amine groups, these particles exhibited excellent ECL behavior in the presence of tris(2.2'-bipyridyl)ruthenium(II). An HBPEI-coated particle possessed an ECL activity that was at least 60 times higher than that of a tripropylamine molecule. Furthermore, these particles could be immobilized on the 3-aminopropyltriethoxysilane-treated quartz substrates to amplify the binding sites for carboxyl groups. Through this approach, PtNPs were applied to the amplified analysis of the hemin/G-quadruplex DNAzyme by using the luminol/H2O2 chemiluminescence method.

Label-Free Colorimetric Detection of Aqueous Mercury Ion (Hg2+) Using Hg2+-Modulated G-Quadruplex-Based DNAzymes

Mercury ion (Hg2+) is able to specifically bind to the thymine-thymine (T-T) base pair in a DNA duplex, thus providing a rationale for DNA-based selective detection of Hg2+ with various means. In this work, we for the first time utilize the Hg2+-mediated T-T base pair to modulate the proper folding of G-quadruplex DNAs and inhibit the DNAzyme activity, thereby pioneering a facile approach to sense Hg2+ with colorimetry. Two bimolecular DNA G-quadruplexes containing many T residues are adopted here, which function well in low- and high-salt conditions, respectively. These G-quadruplex DNAs are able to bind hemin to form the peroxidase-like DNAzymes in the folded state. Upon addition of Hg2+, the proper folding of G-quadruplex DNAs is inhibited due to the formation of T-Hg2+-T complex. Ibis is reflected by the notable change of the Soret band of hemin when investigated by using UV-vis absorption spectroscopy. As a result of Hg2+ inhibition, a sharp decrease in the catalytic activity toward the H2O2-mediated oxidation of 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt (ABTS) is observed, accompanied by a change in solution color. Through this approach, aqueous Hg2+ can be detected at 50 nM (10 ppb) with colorimetry in a facile way, with high selectivity against other metal ions.

G-Quadruplex Aptamers with Peroxidase-Like DNAzyme Functions: Which Is the Best and How Does it Work?

Some G-quadruplex DNA aptamers have been found to strongly bind hemin to form DNAzymes with peroxidase-like activity. To help determine the most suitable DNAzymes and to understand how they work, five previously reported G-quadruplex aptamers were compared for their binding affinity and then the potential catalytic mechanism of their corresponding hemin-G-quadruplex DNAzymes was explored. Among these aptamers, a G-quadruplex named AGRO100 was shown to possess the highest hemin-binding affinity and the best DNAzyme function. This means that AGRO100 is the most ideal candidate for DNAzyme-based analysis. Furthermore, we found the peroxidase-like activity of DNAzyme to be primarily dependent on the concentration of H2O2 and independent of that of the peroxidase substrate (that is, 2,2-azino-bis(3-ethytbenzothiazoline-6-sulfonic acid)diammonium salt). Accordingly, a reaction mechanism for DNAzyme-catalyzed peroxidation is proposed. This study provides new insights into the G-quadruplex-based DNAzymes and will help us to further extend their applications in the analytical field.

Enhanced catalytic DNAzyme for label-free colorimetric detection of DNA

A DNAzyme-based label-free method for the colorimetric detection of DNA is introduced, with a supramolecular hemin G-quartet complex as the sensing element and a 36-mer single-strand DNA as the analyte that is detected at 10 nM.

Aptamer-based label-free method for hemin recognition and DNA assay by capillary electrophoresis with chemiluminescence detection

An aptamer-based label-free approach to hemin recognition and DNA assay using capillary electrophoresis with chemiluminescence detection is introduced here. Two guanine-rich DNA aptamers were used as the recognition element and target DNA, respectively. In the presence of potassium ions, the two aptamers folded into the G-quartet structures, binding hemin with high specificity and affinity. Based on the G-quartet-hemin interactions, the ligand molecule was specifically recognized with a K (d)approximate to 73 nM, and the target DNA could be detected at 0.1 mu M. In phosphate buffer of pH 11.0, hemin catalyzed the H2O2-mediated oxidation of luminol to generate strong chemiluminescence signal; thus the target molecule itself served as an indicator for the molecule-aptamer interaction, which made the labeling and/or modification of aptamers or target molecules unnecessary. This label-free method for molecular recognition and DNA detection is therefore simple, easy, and effective.

Interactions of the human telomeric DNA with terbium-amino acid complexes

The human telomeric DNA can form four-stranded structures: the G-rich strand adopts a G-quadruplex conformation stabilized by G-quartets and the C-rich strand may fold into an I-motif based on intercalated C (.) C+ base pairs. There is intense interests in the design and synthesis of compounds which can target telomeric DNA and inhibit the telomerase activity. Here we report the thermodynamic studies of the two newly synthesized terbium-amino acid complexes bound to the human telomeric G-quadruplex and I-motif DNA which were studied by means of UV-Visible, DNA meltings, fluorescence and circular dichroism. These two complexes can bind to the human telomeric DNA and have shown different features on DNA stability, binding stoichiometry, and sequence-dependent fluorescence enhancement. To our knowledge, this is the first report to show terbium-amino acid complexes can interact with the human telomeric DNA.