阿达玛变换显微图象分析系统测定乳腺肿瘤细胞DNA含量

本文以吖啶橙为细胞DNA荧光探针,探讨了影响阿达玛变换显微图象分析仪定量分析细胞DNA时的细胞荧光强度以及影响人乳腺肿瘤细胞DNA定量分析结果准确性的几个因素,结果表明:(1)乙醇是理想的固定剂,醛类固定剂对细胞A()-DNA复合物的荧光有显著影响;(2)吖啶橙染色液中含Triton X-100时细胞荧光强度明显增大;(3)吖啶橙的最佳荧光染色浓度为50μg/mL;(4)在分析乳腺肿瘤细胞DNA含量(倍性)时,采用外标法(以人血涂片或淋巴结细胞涂上的淋巴细胞为标准二倍体细胞)或内标法(以乳腺肿瘤细胞涂片上的正常上皮细胞为标准二倍体细胞)均能得到较好结果,但后者更为可靠;(5)随机分析六十个以上肿瘤细胞,可得到较好分析结果.     

阿达玛变换显微图象分析系统测定乳腺肿瘤细胞DNA含量

本文以吖啶橙为细胞DNA荧光探针, 探讨了影响阿棕玛显微图象分析仪定量分析细胞DNA时的细胞荧光强度以及影响人乳腺肿瘤细胞DNA定量分析结果准确性的几个因素, 结果表明: (1)乙醇是理想的固定剂, 醛类固定剂对细胞AO-DNA复合物的荧光有显著影响; (2)吖啶橙染色液中含Triton X-100时细胞荧光强芳明显增大; (3)吖啶橙的最佳荧光染色浓度为50μg/mL; (4)在分析乳腺肿瘤细胞DNA含量(倍性)时, 采用外标法(以人血涂片或淋巴结细胞涂上的淋巴细胞为标准二倍体细胞)或内标法(以乳腺肿瘤细胞涂片上的正常上皮细胞为标准二倍体细胞)均能得到较好的结果, 但后者更为可靠; (5)随机分析六十个以上肿瘤细胞, 可得到较好分析结果。     

38例乳腺肿块超声诊断分析

为分析探讨乳腺肿块的高频超声图像特点，提高超声对乳腺肿块的诊断与鉴别水平，利用高频超声探头（７．５￣１０ＭＨｚ）对３８例有手术及病理证实的乳腺肿块进行了诊断，并与病理结果进行对比研究，其中包括有乳腺小叶增生４例，纤维瘤５例，恶性肿瘤１９例。     

阿达玛变换显微图像分析系统在乳腺肿瘤细胞DNA倍性分析…

以正常人外周静脉血淋巴细胞为标准二倍体细胞，以吖橙为细胞ＤＮＡ荧光探针，用阿达玛变换显微图象分析仪测定了冰例乳腺肿瘤的细胞ＤＮＡ倍性与面积呈显著正相关。     

有机锗化合物的离子色谱法分析研究

研究了３种有机锗化合物：β－羧乙基锗倍半氧化物、β－（α－甲基）羧乙基锗倍半氧化物、二－（β－羧乙基）锗氢氧化物的离子色谱法分离和测定条件。以四硼酸钠溶液为淋洗液,硫酸溶液为再生液,梯度淋洗。３种化合物均达到基线分离,检出限分别为０．０３８,０．０３５和０．０２５ｍｇ／Ｌ（以锗计）。方法已用于保健口服液的测定,加标回收率为９５％～１０１％,其结果与氢化物发生－原子荧光光谱法的结果吻合。     

DNA脱碱基位点存在下非氢键配体的增强结合研究

众所周知,插入剂的DNA特性结合位点位于DNA碱基对之间,然而这种非共价相互作用对于含脱碱基（AP）位点的DNA来讲还没有引起足够的重视,虽然在生物细胞中总是存在着DNA脱碱基位点。本文以原黄素（proflavine,PF）为例研究了插入剂对DNA中AP位点的结合特性。实验结果表明,相对于插入位点而言,AP位点是PF的优先结合位点,AP位点的本征结合常数比插入结合常数高一个数量级以上。此外,PF的结合使含脱碱基位点DNA的热稳定性明显提高,表明PF在脱碱基位点的结合构像明显不同于插入结合时的分子定向。本文结果将有助于判断小分子的DNA结合方式所决定的药物的生物化学及生物物理效用。     

二甲基亚砜及一些含氮环化合物与DNA的作用

通过实验探索了将既不溶于水又不能生成可溶性盐的一些含氮稠环化合物溶于二甲基亚砜（DMSO）水溶液中与小牛胸腺DNA作用的途径．溶解实验证明了DMSO及其水溶液对此类化合物极强的溶解性能，DMSO与小牛胸腺作用的紫外光谱（298 K）、圆二色谱（298 K）、~(31)P核磁谱（323 K）和摩尔焓变（298 K）数据均表明：在pH＝7.0的水溶液中，当DMSO浓度小于0．15 mol·L~(-1)时，其与DNA无作用．溶于DMsO水溶液中的八种含氮稠环化合物与小牛胸腺DNA作用的紫外光谱（298 K）、圆二色谱（298 K）和摩尔焓变（298 K）实验结果显示出两种化合物对DNA有较强的嵌插作用．通过对实验结果的分析，本文对具有相同含氮稠环骨架的化合物与DNA嵌播作用的规律和化合物本身微观结构的关系进行了讨论．     

钌（Ⅱ）配合物[Ru（btz）3]2＋和[Ru（btz）（dppz）2]2＋的合成、DNA键合及光断裂研究

本文设计合成了两个含有联噻唑的钌（Ⅱ）配合物[Ru（btz）3]（CIO4）2（1）和[Ru（btz）（dppz）2]（c104）2（2）（btz=4,4’bithiazole,dppz：dipyrido[3,2-a：2’,3’c]phenazine）．通过元素分析、ES-MS、1HNMR和X射线单晶衍射表征了配合物的结构．运用光谱滴定和黏度实验研究了配合物与DNA的作用,发现配合物1与DNA以静电方式结合,配合物2以插入模式与DNA结合．凝胶电泳实验表明配合物1和2在光照射下均可有效断裂DNA,单线杰氢（1O2）和脊某自由墓（OH、县推动断裂反应的活性物种．     

铝离子与脱氧核糖核酸作用的共振光散射研究

在PH2．21的酸性介质中，Al^3 与脱氧核糖核酸（DNA）发生静电作用产生以291．0nm为特征峰的共振光散射（RLS）增强光谱，即Al^3 主要与DNA分子表面的磷酸根结合，但DNA热变性将导致Al^3 与DNA的碱基结合，使光散射信号降低，在291．0nm处的共振光散射（RLS）强度与DNA的浓度呈线性关系，据此建立了用共振光散射测量痕量DNA的新方法，方法的检出限为ng级，用于合成样分析，回收率在91．6％－105．0％。     

基于针状二氧化钛纳米膜的人类肝癌细胞过氧化氢的检测

同济大学化学系田阳课题组研究了基于高导电性针状TiO1纳米膜的细胞色素C（cyt c）直接电子传递，并构筑了新型H2O2传感器，成功地应用于人类肝癌细胞H2O2的检测（Anal．Chem．Doi：1．1021 ac802721x）。研究表明：高导电的针状TiO2纳米膜可以促进了氧化还原酶与电极间的电子传递，采用cyt c作为研究模型，发现其异相电子传递速率常数（ks）为（13．8±2．1）s^-1。     

阿达玛变换显微荧光成像在单细胞定量分析中的应用研究

本文探讨了用阿达玛变换（HT）显微荧光成像系统获得基于256灰度级图像的单细胞荧光强度和解码定标值之间的关系,将归一化的HT图像用于定量分析,建立了适合不同样品的定量分析方法.在此条件下,定量分析数据有很好的准确度和精密度;将系统用于单个乳腺肿瘤细胞的DNA定量分析,对乳腺肿瘤的良恶性及癌变程度进行判断,分析结果与病理学诊断结论一致.     

高分辨阿达玛变换显微荧光图像分析(Ⅰ)——系统的成像能力及其在单个肿瘤细胞分析中的应用

阿达玛变换(Hadamard transform, HT)是一种类似于傅里叶变换的光谱调制技术, 具有多通道同时检测和多通道成像能力. 实现高分辨HT成像的关键在于阿达玛模板的制作, 阿达玛模板有两种, 即移动式机械编码模板(Movable mechanical mask)和固定式光电模板(Stationary electro-optic mask). 在实际成像方面, 移动模板和固定模板各有优缺点: 前者一般用石英玻璃制作, 对光信号不会因模板吸收而导致信号损失, 因此数据很可靠, 而且模板的制作也较为容易, 但由于采用步进电机驱动而容易导致机械故障, 难以实现快速编码; 后者无移动部件, 无机械故障, 因此系统比较紧凑, 但由于它是由液晶材料制成的(可导致信号损失), 从而限制了其在某些光谱区域的使用. 此外, 它对系统的软件设计要求比前者高, 实现高分辨成像更加困难. 正是由于上述原因, 实现快速、高分辨HT成像具有一定难度, 最近有关HT成像技术的报道极少.     

Quantitative DNA imaging in breast tumor cells by a Hadamard transform fluorescence imaging microscope.

This paper presents a novel Hadamard transform (HT) fluorescence imaging microscope by combining multiplexed imaging technique with a conventional upright fluorescence microscope for single-cell imaging and quantitative cellular analysis. The HT imaging microscope can provide 511 x 512-pixel single-cell image with high sensitivity within 21 s. In this study, the high potential value of the microscope in biomedical analysis has been demonstrated by using it to evaluate the malignancy degree of thirty cases of human breast tumors based on the measurements of cellular DNA contents, with conclusions highly accordant with pathological diagnosis. The results show that the HT microscope has the ability to analyze very small specimens and the capability of detecting very high ploidy cells, which are advantages over flow cytometry. The microscope was also successfully applied to cellular morphological analysis, and it was demonstrated that a significant linear relationship exists between tumor nuclear DNA contents and the nuclear area, and malignant and benign tumors are significantly different in both DNA contents and nuclear area. The reliability of the HT microscope in cellular DNA measurements was also investigated.     

Hadamard transform fluorescence image microscopy using one-dimensional movable mask

With a 511-slit one-dimensional (1D) Hadamard mask and a highly sensitive linear charge-coupled device (CCD), spatial multiplexing is performed and a programmable Hadamard transform (HT) microscopic fluorescence imaging system was developed. The system can generate 511×512 pixel format images for small samples. Sensitivity, signal to noise ratio, imaging speed and spatial resolution of this system were discussed. The results show that the system can be applied for single-cell imaging sensitively in a short time. Spatial resolution up to 0.24 μm/pixel, which is close to the resolution limit of the conventional optical microscope, has been obtained under oil lens. The weak native fluorescence imaging for pollen cells can be realized within 1 min. The system has been applied for multi-parameter evaluation of tumor malignancy based on nuclear DNA ploidy measurements for one breast tumor specimen. The result indicates that the system has good application prospect in cell biology and medicine.     

Synthesis, cytotoxicities and DNA-binding affinities of benzofuran-3-ols and their fused analogs

A series of benzofuropyrazoles 2a-i were synthesized in 10-92% from the reaction of 2-aroylbenzofuran-3-ols 1a-i with hydrazine hydrate, and screened for their antitumor activities toward four human solid tumor cell lines, including gastric carcinoma cells MKN45, hepatocellular carcinoma cells HepG2, breast cancer cells MCF-7, and lung cancer cells A549. The results indicated that both compounds 1a-i and 2a-i displayed moderate antitumor activities. Among them, compound 2e exhibited potent inhibitory activity toward all the four tumor cell lines. In addition, compounds 1e and 2e showed strong DNA-binding affinities, and induced an increase in the viscosity of calf-thymus DNA, suggesting that they might act as an intercalator.     

DNA-based electrochemical biosensors for monitoring of bis-indoles as potential antitumoral agents, chemistry, X-ray crystallography

Facile and practical electrochemical DNA bioassay, X-ray diffraction analysis, synthesis and 1H and 13C NMR data of the 5,5'-disubstituted-3,3'-methanediyl-bis-indoles are reported. On the basis of electrochemical measurements we have hypothesized that the analyzed bis-indoles have an effect on human tumor cells due to DNA binding at adenine-thymidine deoxynucleotides rich region in a concentration/substituent dependent manner. Interesting N-H...pi and hydrogen-bonding intermolecular interactions were observed which may differentiate their biological features. The 5,5'-dimethoxy-3,3'-methanediyl-bis-indole (2) was found to reduce considerably the growth of cancer cell lines HOP-92 (lung), A498 (renal) and MDA-MB-231/1TCC (breast). The results indicate that title compounds could be interesting as potential antitumoral chemotherapeutics.     

Serological proteome analysis of dogs with breast cancer unveils common serum biomarkers with human counterparts

Canine mammary tumor is being touted as a model for investigating the human breast cancer. Breast cancer of the both species has similar biological behavior, histopathologic characteristics, and metastatic pattern. In this study, we used the serological proteome analysis to detect autoantigens that elicit a humoral response in dogs with mammary tumor in order to identify serum biomarkers with potential usefulness as diagnostic markers and to better understand molecular mechanisms underlying canine breast cancer development. Protein extract from a cell line was subject to 2DE followed by Western blotting using sera from 15 dogs with mammary tumor and sera from 15 healthy control dogs. Immunoreactive autoantigens were subsequently identified by the MALDI‐TOF MS. Four autoantigens, including manganese‐superoxide dismutase, triose phosphate isomerase, alpha‐enolase, and phosphoglycerate mutase1, with significantly higher immunoreactivity in the tumor samples than in the normal samples were identified as biomarker candidates. Immunohistochemistry and Western blotting revealed higher expression of these biomarkers in the malignant tumors than in the normal or benign tumors. The autoantigens found in this study have been reported to elicit autoantibody response in the human breast cancer, indicating the similarity of breast cancer proteome profile in dogs with that in human beings.     

Discovery of Cell‐Permeable Inhibitors That Target the BRCT Domain of BRCA1 Protein by Using a Small‐Molecule Microarray

BRCTs are phosphoserine‐binding domains found in proteins involved in DNA repair, DNA damage response and cell cycle regulation. BRCA1 is a BRCT domain‐containing, tumor‐suppressing protein expressed in the cells of breast and other human tissues. Mutations in BRCA1 have been found in ca. 50 % of hereditary breast cancers. Cell‐permeable, small‐molecule BRCA1 inhibitors are promising anticancer agents, but are not available currently. Herein, with the assist of microarray‐based platforms, we have discovered the first cell‐permeable protein–protein interaction (PPI) inhibitors against BRCA1. By targeting the (BRCT)₂ domain, we showed compound 15 a and its prodrug 15 b inhibited BRCA1 activities in tumor cells, sensitized these cells to ionizing radiation‐induced apoptosis, and showed synergistic inhibitory effect when used in combination with Olaparib (a small‐molecule inhibitor of poly‐ADP‐ribose polymerase) and Etoposide (a small‐molecule inhibitor of topoisomerase II). Unlike previously reported peptide‐based PPI inhibitors of BRCA1, our compounds are small‐molecule‐like and could be directly administered to tumor cells, thus making them useful for future studies of BRCA1/PARP‐related pathways in DNA damage and repair response, and in cancer therapy.