在体检测γ-氨基丁酸的MEGA-PRESS序列优化设计

γ-氨基丁酸（γ-aminobutyric acid,GABA）含量异常会引发一系列大脑疾病,因此在体检测人脑GABA含量成为一种十分必要的医疗技术.本文基于GABA检测常用的谱编辑方法MEGA-PRESS,引入性能更好的SLR脉冲代替常用的高斯脉冲,并使用单频带和双频带结合的方法激发特定频率的物质,在联影（United Imaging Healthcare,UIH）的uMR780磁共振系统上实现了一种优化的MEGA-PRESS序列.结果表明：SLR脉冲可以减少大分子（macromolecule,MM）信号对GABA信号的干扰;单频带和双频带结合可以更好地抑制残余水信号对GABA^*信号的影响.水模实验和人体实验结果均证实优化的MEGA-PRESS序列可以更好地实现GABA^*含量在体检测.     

基于i-motifDNA构型变化和纳米金紫外-可见吸收光谱的pH测定研究

利用纳米金对DNA构型的区分效应设计了一种快速灵敏可视化的pH计。对i-motif DNA-纳米金体系的紫外可见吸收光谱变化进行了系统研究,考察了盐浓度、i-motif DNA浓度、四重体形成时间以及DNA序列对pH响应的影响。在既定条件下,纳米金的紫外-可见吸收光谱在pH 5.3～7.0范围内呈现规律性变化,在520nm处的吸光度逐渐增大,在700nm处的吸光度逐渐减小,纳米金的颜色逐渐由蓝紫色变化至红色。该pH计具有纳米金无需修饰、可视化、成本低、速度快等特点,有望用于某些pH值发生变化的生命过程的监测。     

Recent advances in rice genome and chromosome structure research by fluorescence in situ hybridization (FISH)

Fluorescence in situ hybridization (FISH) is an effective method for the physical mapping of genes and repetitive DNA sequences on chromosomes. Physical mapping of unique nucleotide sequences on specific rice chromosome regions was performed using a combination of chromosome identification and highly sensitive FISH. Increases in the detection sensitivity of smaller DNA sequences and improvements in spatial resolution have ushered in a new phase in FISH technology. Thus, it is now possible to perform in situ hybridization on somatic chromosomes, pachytene chromosomes, and even on extended DNA fibers (EDFs). Pachytene-FISH allows the integration of genetic linkage maps and quantitative chromosome maps. Visualization methods using FISH can reveal the spatial organization of the centromere, heterochromatin/euchromatin, and the terminal structures of rice chromosomes. Furthermore, EDF-FISH and the DNA combing technique can resolve a spatial distance of 1 kb between adjacent DNA sequences, and the detection of even a 300-bp target is now feasible. The copy numbers of various repetitive sequences and the sizes of various DNA molecules were quantitatively measured using the molecular combing technique. This review describes the significance of these advances in molecular cytology in rice and discusses future applications in plant studies using visualization techniques.     

Wavelet-based multifractal analysis of DNA sequences by using chaos-game representation

Chaos game representation (CGR) is proposed as a scale-independent representation for DNA sequences and provides information about the statistical distribution of oligonucleotides in a DNA sequence. CGR images of DNA sequences represent some kinds of fractal patterns, but the common multifractal analysis based on the box counting method cannot deal with CGR images perfectly. Here, the wavelet transform modulus maxima (WTMM) method is applied to the multifractal analysis of CGR images. The results show that the scale-invariance range of CGR edge images can be extended to three orders of magnitude, and complete singularity spectra can be calculated. Spectrum parameters such as the singularity spectrum span are extracted to describe the statistical character of DNA sequences. Compared with the singularity spectrum span, exon sequences with a minimal spectrum span have the most uniform fractal structure. Also, the singularity spectrum parameters are related to oligonucleotide length, sequence component and species, thereby providing a method of studying the length polymorphism of repeat oligonucleotides.     

DNA Biosensor with High Sensitivity Amplified by Gold Nanoparticles

A biosensor based on quartz crystal microbalance (QCM) using 50 nm gold nanoparticles as the amplification probe for DNA detection was reported in this paper. It had been found that a DNA detection sensitivity of 10^–14 M could be obtained, which was higher than what has been ever reported using the same method. In the curve of weight increase (m) against the target DNA in solution, a plateau appeared as the concentration of analyte DNA increasing, implying the existence of monolayer of gold nanoparticles binding on the top of target DNA. The high sensitivity was explained not only by the weight of the larger particles, but also the larger area occupied by the larger particles which need less target DNA for their binding.     

Automated detection system of single nucleotide polymorphisms using two kinds of functional magnetic nanoparticles

Single nucleotide polymorphisms (SNPs) comprise the most abundant source of genetic variation in the human genome wide codominant SNPs identification. Therefore, large-scale codominant SNPs identification, especially for those associated with complex diseases, has induced the need for completely high-throughput and automated SNP genotyping method. Herein, we present an automated detection system of SNPs based on two kinds of functional magnetic nanoparticles (MNPs) and dual-color hybridization. The amido-modified MNPs (NH₂-MNPs) modified with APTES were used for DNA extraction from whole blood directly by electrostatic reaction, and followed by PCR, was successfully performed. Furthermore, biotinylated PCR products were captured on the streptavidin-coated MNPs (SA-MNPs) and interrogated by hybridization with a pair of dual-color probes to determine SNP, then the genotype of each sample can be simultaneously identified by scanning the microarray printed with the denatured fluorescent probes. This system provided a rapid, sensitive and highly versatile automated procedure that will greatly facilitate the analysis of different known SNPs in human genome.     

DNA微阵列制备的新进展

随着人类基因组测序的完成和蛋白组学工程的开展，具有高产出测序的特性的DNA微阵列技术的发展日新月异，其应用已经深入到了生命科学研究的很多方面．与此同时，几种新的DNA微阵列制备技术迅速发展起来，文章介绍了滚轮放大技术、聚酰胺胺表面法、三甲氧基对胺苯基硅烷／重氮化法、化学纳米印迹法等四种制备方法的新进展。     

两性离子表面活性剂BS-12与DNA作用的共振光散射光谱

直接将两性离子表面活性剂用于DNA的共振光散射测定。在pH 9.3的缓冲液中,DNA与十二烷基二甲基甜菜碱(BS-12)在388 nm处有共振光散射增强,其强度与核酸的浓度呈线性关系,据此建立了一种测定DNA的共振光散射法。该方法具有操作简单、线性范围宽、检出限低的特点。fsDNA与ctDNA的线性范围分别为0.25～12.0 μg·mL-1和0.25～11.0 μg· mL-1,检测限分别为0.15和0.16 ng·mL-1。该法用于混合样品中DNA的测定,结果满意。     

DNA分子链电子结构特性研究

在单电子紧束缚近似下，建立了一维无序二元DNA分子链模型，计算了链长为2×10⁴个碱基对的DNA分子链的电子态密度、局域化特性，并探讨了碱基对的不同组分、格点能量无序度对电子局域态的影响.结果表明：由于DNA分子链中格点能量无序及碱基对的不同组分的存在，其电子波函数呈现出局域化的特性，而局域长度作为衡量电子局域化程度的一个尺度，受碱基对的组分及格点能量无序度的影响. 关键词： DNA分子链 电子结构 电子局域态 局域长度     

Fluorescent Biosensor Based on Hairpin DNA Stabilized Copper Nanoclusters for Chlamydia trachomatis Detection

Chlamydia trachomatis (C. trachomatis) is a kind of intracellular parasitic microorganism, which can causes many diseases such as trachoma. In this strategy, a specific hairpin DNA with the probe loop as specific regions to recognize C. trachomatis DNA with strong affinity was designed, and its stem consisted of 24 AT base pairs as an effective template for hairpin DNA-CuNCs formation. In the absence of C. trachomatis DNA, the detection system showed strong orange fluorescence emission peaks at 606 nm. In the presence of C. trachomatis DNA, the conformation of DNA probe changed after hybridizing with C. trachomatis DNA. Then, the amount of hairpin DNA-CuNCs was reduced and resulted in low fluorescence emission. C. trachomatis DNA displayed a significant inhibitory effect on the synthesis of fluorescent hairpin DNA-CuNCs due to the competition between C. trachomatis DNA and the specific hairpin DNA. Under the optimal experimental conditions, different concentrations of C. trachomatis were tested and the results showed a good linear relationship in the range of 50 nM to 950 nM. Moreover, the detection limit was 18.5 nM and this detection method possessed good selectivity. Finally, the fluorescent biosensor had been successfully applied to the detection of C. trachomatis target sequence in HeLa cell lysate, providing a new strategy for the detection of C. trachomatis.

     

基于铜离子（Ⅱ）类过氧化物酶性质可视化比色法检测过氧化氢

过氧化氢（H₂O₂）在食品工业、环境监测分析、燃料电池、临床诊断等领域得到十分广泛的应用。过氧化氢不仅是严重疾病的生物标志物,也是重要的食品添加剂。无论从食品安全还是人类健康考虑,对过氧化氢构建简便、快速、灵敏的检测方法具有重要意义。比色法因具有易操作、成本低及检测结果可视化等优点被广泛关注。比色法普遍使用的酶是辣根过氧化物酶(HRP)。但天然酶存在易失活、生产成本高及稳定性差等不足。纳米酶克服了HRP易失活的缺点。然而部分纳米酶合成复杂、需要表征、水溶性差及催化活性低等。与HRP和纳米酶相比,具有类过氧化物酶性质的铜离子（Ⅱ）,不仅具有灵敏度高的特点,而且不需要复杂的合成、易获得、易储存、不需修饰可直接使用、操作简单及分析成本低廉等优势。铜离子（Ⅱ）类酶能够通过催化过氧化氢生成羟基自由基,氧化3,3’,5,5’-四甲基联苯胺（TMB）而产生氧化态TMB,使溶液的颜色由无色逐渐转变为蓝色,从而产生光信号,这些光信号可以肉眼识别和通过紫外分光光度计检测。基于上述原理构建过氧化氢的快速比色传感并应用于银耳样品的检测。实验考察了体系中过氧化氢浓度、pH值、温度等因素对吸光度的影响。选择缓冲溶液pH值为3.0,温度40 ℃,TMB和铜离子（Ⅱ）浓度分别为6.0×10-4和8.0×10-3 mol·L-1,反应时间为20 min。在最优条件下,过氧化氢浓度与体系的吸光度值呈良好的线性关系,线性范围是0.08~40 μmol·L-1,检测限为0.14 μmol·L-1。该法可检测银耳中过氧化氢的浓度,加标回收率在97.10%~107.08%之间,相对标准偏差（RSD）小于5%。该研究无需特殊检测设备条件下实现过氧化氢的简单、快速、成本低廉且灵敏的可视化检测,有利于过氧化氢的定量检测在食品领域和临床上的快速推广。     

Thiol-Capped Gold Nanoparticle Biosensors for Rapid and Sensitive Visual Colorimetric Detection of <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis>

In the last few years, gold nanoparticle biosensors have been developed for rapid, precise, easy and inexpensive with high specificity and sensitivity detection of human, plant and animal pathogens. Klebsiella pneumoniae serotype K2 is one of the common gram-negative pathogens with high prevalence. Therefore, it is essential to provide the effective and exclusive method to detect the bacteria. Klebsiella pneumoniae serotype K2 strain ATCC9997 genomic DNA was applied to establish the detection protocol either with thiol-capped oligonucleotide probes and gold nanoparticles or polymerase chain reaction based on K2A gene sequence. In the presence of the genomic DNA and oligonucleotide probes, a change in the color of gold nanoparticles and maximum changes in wavelength at 550-650 nm was achieved. In addition, the result showed specificity of 15?×?10⁵ CFU/mL and 9 pg/μL by gold nanoparticles probes. The lower limit of detection obtained by PCR method was 1 pg/μL. Moreover, results demonstrated a great specificity of the designed primers and probes for colorimetric detection assay and PCR. Colorimetric detection using gold nanoparticle probe with advantages such as the lower time required for detection and no need for expensive detection instrumentation compared to the biochemical and molecular methods could be introduced for rapid, accurate detection of the bacteria.     

表面增强拉曼光谱技术快速检测神经性化学毒剂模拟剂

神经性化学毒剂具有毒性高、挥发性好和作用快等特点,在浓度很低的情况下就可以造成很大的伤亡,因此成为现场快速检测的难点。而化学毒剂沙林（甲氟膦酸异丙酯,GB）是一种经常用的军事神经性毒剂,该毒剂可通过抑制乙酰胆碱酯酶来破坏神经系统的功能,同时该毒剂被吸入后在人体内的降解速度很慢。为了避免和降低该类毒剂的污染和对人员伤害,迫切需要发展一种灵敏度高、准确性好、响应时间短和可便携化的检测技术检测GB。表面增强拉曼光谱(SERS)检测方法具有灵敏度高、操作简单和响应速度快的特点,成为检测水中痕量化学毒剂的有效方法之一。将购买的Ag纳米溶胶进行离心,然后将其组装固定在硅基Au膜表面从而制备高SERS增强的基底来对化学毒剂模拟剂甲基磷酸二甲酯（DMMP）的快速检测方法进行研究。在实验中,通过优化团聚剂的离子强度、测试方法等来对检测条件进行优化筛选。通过对比不同的团聚剂HCl,KI,MgSO₄,NaCl和NaOH,最终得到最优的离子强度,从而确定1 mol·L-1的KI为团聚剂时具有最好的效果。分别对不同的检测方法来进行了相应的优化,通过对比芯片法和液态溶胶法,最终发现改进后的芯片法能够获得较好的检测效果。最终确定的检测方法为将1 mol·L-1的KI和待测溶液（DMMP）混匀,然后滴在事先准备的以Ag纳米溶胶为基质制得的SERS芯片上,用波长为785 nm激光的便携式拉曼光谱仪直接进行检测,最低可以测至10 μg·L-1。而在文献资料中报道美军短期（＜7 d）饮用水最大暴露安全指南规定对神经性化学毒剂最低检出限是10 μg·L-1,因此采用该SERS检测方法,满足了军队应对化学战或者恐怖袭击的行动的需要。实验结果表明该方法突破了便携拉曼光谱仪灵敏度低的局限,解决了痕量神经性毒剂现场快速检测难题,拓展了SERS技术在化学侦察领域的应用。     

基于RRS的人血清白蛋白快速检测系统设计

在临床中人血清白蛋白浓度的快速检测对多种重大疾病的诊断具有重要意义,而现有的医疗检测设备普遍存在体积大、测试周期长、操作复杂等缺点,设计研究一种快速的、高精度的便携式人血清白蛋白浓度检测系统具有非常广阔的应用前景。设计了基于共振瑞利散射光谱分析的人血清白蛋白浓度检测系统。由于人血清白蛋白与四氨基酞菁铜反应产生的溶液在475.0 nm处具有共振瑞利散射增强的效果,从而构建了共振瑞利散射强度与人血清白蛋白浓度的函数。实验采用紫外半导体激光器与475.0 nm窄带滤光片联用,配合高增益光电探测器采集散射光信号。采用不同浓度的四氨基酞菁铜溶液分别对不同浓度的人血清白蛋白进行检测,实验结果显示,响应电压会随人血清白蛋白浓度的提高而明显增大,而对四氨基酞菁铜的浓度变化并不敏感;散射光产生的响应电压与人血清白蛋白浓度在0.1～1.0 μg·mL-1范围内基本满足线性变化,满足快速检测的设计要求。     

Target detection for low cost uncooled MWIR cameras based on empirical mode decomposition

In this work, a novel method for detecting low intensity fast moving objects with low cost Medium Wavelength Infrared (MWIR) cameras is proposed. The method is based on background subtraction in a video sequence obtained with a low density Focal Plane Array (FPA) of the newly available uncooled lead selenide (PbSe) detectors. Thermal instability along with the lack of specific electronics and mechanical devices for canceling the effect of distortion make background image identification very difficult. As a result, the identification of targets is performed in low signal to noise ratio (SNR) conditions, which may considerably restrict the sensitivity of the detection algorithm. These problems are addressed in this work by means of a new technique based on the empirical mode decomposition, which accomplishes drift estimation and target detection. Given that background estimation is the most important stage for detecting, a previous denoising step enabling a better drift estimation is designed. Comparisons are conducted against a denoising technique based on the wavelet transform and also with traditional drift estimation methods such as Kalman filtering and running average. The results reported by the simulations show that the proposed scheme has superior performance.     

纺织品中邻苯二甲酸酯的表面增强拉曼光谱快速检测

随着绿色纺织理念的不断深入,国际上对于纺织品中的有毒有害化学品越来越重视。纺织品中常用的邻苯二甲酸酯(PAEs)具有生殖毒性、致突变和致癌性,可通过空气、水、食物三大途径进入人体,干扰人体的内分泌系统。由于PAEs对生态系统和公共卫生环境潜在不利的影响,近年来引起越来越多人们的关注。目前,检测PAEs的方法主要是色谱法和色-质联用法,这些方法虽然灵敏度高,但是存在着前处理繁琐复杂,耗时久,检测成本高,需要专业技术人员等缺点,不适合生产过程中的快速分析。而其他方法如,酶联免疫法等研究较少,且存在样品基质干扰,易出现假阳性等问题。因此,建立纺织品中邻苯二甲酸酯的快速分析技术具有重要意义。表面增强拉曼光谱(SERS)作为一种分子振动光谱可提供丰富的分子结构信息,具有极高的灵敏度,广泛应用于食品安全、环境监测和国家安全等领域。研究中提出并建立了一种结合便携式拉曼光谱仪,利用SERS实现纺织品中邻苯二甲酸酯的快速定量检测方法。首先利用水合肼将非水溶性的邻苯二甲酸酯类化合物转化为水溶性的邻苯二甲酰肼。同时,利用纳米金溶胶作为SERS基底,使转化后的邻苯二甲酰肼吸附于金溶胶表面,从而实现其拉曼信号的放大与检测。结果表明,通过这种方法,可实现多种邻苯二甲酸酯的快速检测。进一步研究还表明,在5~150 mg·L^-1范围内,邻苯二甲酸酯浓度与其拉曼光谱强度呈线性关系,线性方程为Y=139.04X+5 465.32,相关系数为0.993 0,检出限为5 mg·L^-1。利用该方法,还实现了不同纺织品中多种邻苯二甲酸酯的快速检测,加标回收率达80%以上,且不受纺织品中其他成分的干扰。所建立表面增强拉曼光谱检测方法操作简便、成本低且结果准确,适用于纺织品中邻苯二甲酸酯类增塑剂的快速定量检测。     

A novel chaotic based image encryption using a hybrid model of deoxyribonucleic acid and cellular automata

Currently, there are many studies have conducted on developing security of the digital image in order to protect such data while they are sending on the internet. This work aims to propose a new approach based on a hybrid model of the Tinkerbell chaotic map, deoxyribonucleic acid (DNA) and cellular automata (CA). DNA rules, DNA sequence XOR operator and CA rules are used simultaneously to encrypt the plain-image pixels. To determine rule number in DNA sequence and also CA, a 2-dimension Tinkerbell chaotic map is employed. Experimental results and computer simulations, both confirm that the proposed scheme not only demonstrates outstanding encryption, but also resists various typical attacks.     

快速简便的杂交检测DNA

Southern杂交作为黄金标准,已广泛运用于DNA的检测上。但是,经典的southern杂交和近年来一些DNA检测方法存在放射性污染,操作繁琐,耗时,对实验仪器设备要求高等问题。本文利用异硫氰酸荧光素(FITC)标记的dUTP合成DNA探针建立了快速检测DNA的液相杂交方法。该方法包括探针制备、液相杂交、电泳分离和信号检测四个步骤,并在此基础上对FITC-双链和FITC-单链探针的杂交效果作了比较。结果显示,使用FITC标记的两种探针都能取得良好的实验结果,但单链探针较双链的检测灵敏度高;双链DNA探针可以检测出0.8 μg(3.64×10-13 mol)的质粒,而单链DNA探针可以检测出0.38 μg(1.82×10-13 mol)的质粒DNA,在检出效率上是前者的2.1倍。整个检测过程操作简便,可在3 h内完成,可较好地解决了其他DNA检测方法存在的费时费力的问题。