904 resultados para MYCN-AMPLIFICATION
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Quantum-dot laser diodes (QD-LDs) with a Fabry-Perot cavity and quantum-dot semiconductor optical amplifiers (QD-SOAs) with 7° tilted cavity were fabricated. The influence of a tilted cavity on optoelectronic active devices was also investigated. For the QD-LD, high performance was observed at room temperature. The threshold current was below 30 mA and the slope efficiency was 0.36 W/A. In contrast, the threshold current of the QD-SOA approached 1000 mA, which indicated that low facet reflectivity was obtained due to the tilted cavity design.A much more inverted carrier population was found in the QD-SOA active region at high operating current, thus offering a large optical gain and preserving the advantages of quantum dots in optical amplification and processing applications. Due to the inhomogeneity and excited state transition of quantum dots, the full width at half maximum of the electroluminescence spectrum of the QD-SOA was 81.6 nm at the injection current of 120 mA, which was ideal for broad bandwidth application in a wavelength division multiplexing system. In addition, there was more than one lasing peak in the lasing spectra of both devices and the separation of these peak positions was 6-8 nm,which is approximately equal to the homogeneous broadening of quantum dots.
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We have experimentally demonstrated pulses 0.4 mJ in duration smaller than 12 fs with an excellent spatial beam profile by self-guided propagation in argon. The original 52 fs pulses from the chirped pulsed amplification laser system are first precompressed to 32 fs by inserting an acoustic optical programmable dispersive filter instrument into the laser system for spectrum reshaping and dispersion compensation, and the pulse spectrum is subsequently broadened by filamentation in an argon cell. By using chirped mirrors for post-dispersion compensation, the pulses are successfully compressed to smaller than 12 fs.
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Slow-light effects in photonic crystal (PC) waveguides can enhance light-mater interaction near the photonic band edge, which can be used to design a short cavity length semiconductor optical amplifier (SOA). In this paper, a novel SOA based on slow-light effects in PC waveguides (PCSOA) is presented. To realize the amplification of the optical signal with polarization independence, a PCSOA is designed with a compensated structure. The cascaded structure leads to a balanced amplification to the TE and TM polarized light.
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研究背景与目的:近二十年来,抗生素的广泛使用以及一些不当应用导致临床上出现大量的耐药性病原菌,所以不易产生耐药性的抗菌肽就成为目前研究的热点。本课题组此前的研究表明无指盘臭蛙(Odorrana grahami)皮肤抗菌肽具有广谱抗菌活性,但对真核细胞没有毒性,因此有成为新型药物的潜力。本研究采用毕赤酵母真核表达系统来生物合成抗菌肽Odorgrin A和Odorgrin C,为大量获取抗菌肽资源提供技术支撑。 方法:依照Odorgrin A和C的氨基酸序列、采用酵母偏爱密码子分别设计并化学合成了相应的目的基因序列。目的片段从合成质粒上用Xho Ι和EcoR Ι双酶切下后,与经同样限制酶完全酶切pPIC9K载体所获得的两个大片段直接连接,并转化至大肠杆菌DH5α。用PCR扩增、酶切及测序检测,鉴定正确的重组质粒。提取大量表达载体pPIC9K - Odo A和C并使之线性化后经电击法分别转化毕赤酵母(Pichia pastoris)GS115宿主菌,用营养缺陷型筛选、遗传霉素抗性筛选、PCR扩增和测序检测,鉴定并筛选出对G418具高抗性的Odorgrin A和C重组酵母菌。用甲醇对之进行诱导表达,SDS - PAGE电泳及反相层析检测表达产物,并做抑菌活性检测。 成果:PCR扩增、酶切及测序等结果表明表达载体pPIC9K - Odo A和C构建成功。营养缺陷型筛选、遗传霉素抗性筛选、PCR扩增和测序等证实pPIC9K - Odo A和C已整合入酵母基因组中。SDS - PAGE电泳及反相层析结果表明抗菌肽Odorgrin A和C成功地获得了分泌表达。而抑菌活性实验则检测到部分阳性克隆菌诱导分泌表达的抗菌肽Odorgrin A和C都对测试菌的生长具有较高(>94%)的抑制率。 结论:无指盘臭蛙皮肤抗菌肽Odorgrin A和Odorgrin C基因的表达载体都构建成功,并且都在毕赤酵母系统中获得了成功表达。 Background & Objective: In the recent twenty years, a lot of pathogenic bacteria have come forth in clinic with durable trait derived from making use of and abusing the traditional antibiotics. Therefore, studying antimicrobial peptides, not be easy to be invalidated by durable bacteria, are becomimg popular and important. The skin antimicrobial peptides of Odorrana grahami with broad spectrum antibacterial activity and no toxicity to eukaryotic cell, discovered by previous research work of our workgroup, are looked forward to being potential medication. Pichia pastoris expressional system was used for biosynthesis antimicrobial peptides Odorgrin A and Odorgrin C in this study, for producing abundant antimicrobial peptides. Methods: The foreign fragments which included Odorgrin A or Odorgrin C gene according to their amino acid sequence respectively were synthesized based on the biased codon usage of yeast. The DNA fragments, obtained from the plasmids containing them by digested with Xho Ι and EcoR Ι, were directly ligated with the two bigger fragments obtained from the vector pPIC9K by digested with the same restriction enzymes. And then they were transformed into Escherichia coli DH5α to be selected and amplified positive colonies. The recombinants were testified by using PCR amplification, enzymes digestion and sequencing of the foreign fragment. After the expressional vector pPIC9K - Odo A and pPIC9K - Odo C were linearized, they were transformed into Pichia pastoris GS115 strain by the electroporation. Then the positive colonies which were of the highest geneticin resistant were selected through auxotrophic screening, genetic resistant screening, PCR amplification and sequencing of the inserted fragment. Methanol was used to induce the recombinant yeasts to express the foreign gene. SDS-PAGE electrophoresis, reversed phase chromatography and antibacterial activity experiment were used to testify the expressional products. Results: The evidences of PCR, enzymes digestion and sequence analysis confirmed that the expressional vector pPIC9K - Odo A and pPIC9K - Odo C have been constructed correctly. The results of auxotrophic screening, of genetic resistant screening, of PCR and sequencing of the foreign fragment showed that Odorgrin A and Odorgrin C gene have been homologous integrated with the Pichia pastoris genome. And it was also testified that antimicrobial peptides Odorgrin A and Odorgrin C have been expressed successfully by using SDS - PAGE electrophoresis, reversed phase chromatography and antibacterial activity experiment. Conclusion: The expressional vector of the skin antimicrobial peptides Odorgrin A and Odorgrin C gene of Odorrana grahami have been constructed correctly and both of the genes have been expressed successfully in Pichia pastoris system in this study.
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糯小麦在食品加工业、淀粉加工业及其它工业上有着重要用途,是近年来许多国家小麦研究的重要课题。国外糯小麦选育尚未突破高产与糯性相结合的难点,国内目前还没有培育出高蛋白强筋型的糯小麦品种,这在一定程度上与缺乏合适的育种方法和高效、实用的糯小麦分子标记辅助育种技术有关。国内外对Wx基因效应的研究主要利用缺体-四体系、重组自交系或近等基因系,还未见有利用遗传背景相同的BC5F2代回交改良群体的报道。 糯性位点近等基因系是小麦淀粉品质育种的重要材料,而我国目前还没有一套中国栽培小麦遗传背景的糯性位点近等基因系。为了选育部分糯小麦、全糯小麦和中国栽培小麦遗传背景的糯性位点近等基因系,我们利用Wx蛋白电泳和高效实用的分子标记技术体系来鉴定糯小麦杂交后代的基因型,结果证明该体系能有效地用于糯小麦的分子标记辅助育种。以中国春糯性位点全套近等基因系为研究材料,对小麦Wx基因的6个STS标记和1个CAPS标记进行了筛选,改良PCR扩增条件以及产物检测方式后,从这些标记中筛选出3个标记,包括鉴定Wx-A1、Wx-D1位点的2个共显性STS标记和Wx-B1位点的1个显性STS标记。利用上述3个分子标记从BC5F2 代回交改良群体中筛选出了8种Wx基因型,经卡方检验,其分离比符合3对基因的分离比例,其中基因型为aabbdd的植株有2株,直链淀粉含量分别为1.81%和0.82%,为全糯小麦;基因型为AAbbdd,aabbDD的部分糯性植株各有1株,直链淀粉含量分别为15.24%和17.57%。以上4株植株的农艺性状和品质性状接近回交亲本“川育12”,并明显优于全糯材料“98Y1441”,表明采用回交法与Wx基因分子标记辅助选择相结合,有助于培育高产、优质的全糯和部分糯小麦。同时,本研究中建立的分子标记技术体系,也为选育具有中国栽培小麦遗传背景的糯性位点近等基因系奠定了基础。 在基因型鉴定的基础上,利用糯小麦杂交后代BC5F2代回交改良群体研究了各基因缺失降低直链淀粉含量的效果和各基因合成直链淀粉的能力,以及直链淀粉含量与农艺性状、品质性状、淀粉糊化特性等的相关性。缺失不同Wx基因的8种基因型,直链淀粉含量差异显著。研究单缺失基因型发现,减少效应最大的是Wx-B1b基因,Wx-B1b和Wx-D1b基因没有显著差异,减少效应最小的是Wx-A1b基因。研究双缺失基因型发现,Wx基因合成直链淀粉的能力,Wx-B1a基因最高,Wx-A1a基因最低,而Wx-B1a和Wx-D1a基因差异很小;直链淀粉含量与株高、穗长、小穗数、穗粒数、千粒重等农艺性状相关不显著,表明淀粉品质育种可以与高产育种实现有机结合;直链淀粉含量与SDS-沉降值呈显著负相关(r=-0.726),说明直链淀粉含量降低在一定程度上有利于提高小麦营养与加工品质,这一结果至今未见有文献报道;全糯类型的淀粉糊化特性与其他类型显著不同,具有最高的峰值粘度和稀懈值,最低的低谷粘度、最终粘度、反弹值、峰值时间、糊化温度、起始糊化温度,表明糯小麦淀粉在食品和工业上具有特殊用途;稀懈值与直链淀粉含量呈极显著负相关(r=-0.969),其他粘度参数与直链淀粉含量呈显著正相关(最终粘度r=0.797,低谷粘度r=0.910、反弹值r=0.954、峰值时间r=0.970、糊化温度r=0.962、起始糊化温度r=0.932)。以上结论可为糯小麦品种选育和淀粉品质改良提供理论依据。 Waxy wheat is very important in food processing industry, starch processing industry and the other industries, so it is a focus of wheat research in many countries these years. Foreign wheat breeders have not conquered the difficulty of high yield combined with waxy character, and there is no waxy wheat variety with high protein and strong gluten in China at present,all of which were caused by lacking proper breeding methods and effective, applied molecular markers-assisted selection technique at a certain extent. Until now, research about the effect of waxy genes mainly depended on nullisomic-tetrasomic lines, recombinant lines or near-isogenic lines, and it is lacking the reports of using improved BC5F2 backcross progenies under the common genetic background. Near-isogenic lines at the Wx loci are important materials for wheat starch quality breeding. However, there are no such lines under the Chinese cultivated wheat genetic background. To develop partial waxy wheats, waxy wheats and near-isogenic lines at the Wx loci under the Chinese cultivated wheat genetic background, we use SDS-PAGE of waxy proteins and effective, applied molecular marker-assisted selection technical system to identify the genotype of waxy wheat’s progenies. The results indicated that such a system is applicable in waxy wheat’s molecular marker-assisted selection effectively. A series of Chinese Spring Wx loci near-isogenic lines were used to identify the specific bands of six STS markers and one CAPS marker of Wx genes. After optimizing PCR amplification and separating of PCR products, three co-dominant and dominant STS-markers were identified at the Wx-A1, Wx-D1 and Wx-B1 loci, respectively, which were used to identify the genotype of waxy wheat’s progenies. Eight Wx genotypes were developed from the improved BC5F2 backcross progenies, which follows Mendelian segregation. Among them, there were two aabbdd waxy plants whose amylose content in the BC5F3 seeds were 1.81% and 0.82%, respectively. In addition, there were partial waxy plants (AAbbdd and aabbDD) whose amylose content in the BC5F3 seeds were 15.24 % and 17.57%, respectively. Most agronomic and quality traits of these four plants resembled those of the recurrent parent “Chuanyu 12”, and superior to waxy wheat parent “98Y1441”. This shows that backcross approach in combination with molecular marker-assisted selection of waxy genes is helpful to develop partial and full waxy wheat with good traits in the waxy wheat breeding program. At the same time, molecular marker-assisted selection technical system in this paper, also establish the base for breeding the near-isogenic lines at the Wx loci under the Chinese cultivated wheat genetic background. According to the results of genotype identification, we use waxy wheat’s improved BC5F2 backcross progenies to verify the effects of null alleles on reducing amylose content and determine the amylose synthesis capacity of each Wx gene independently, and investigate the relativity among amylose content with agonomic traits, quality traits, starch pasting properties respectively. There was significant difference in amylose content of the eight genotypes carrying different null Wx alleles. The reducing effect of the single null alleles was the most significant in Wx-B1b, followed by Wx-D1b and Wx-A1b, and there was no significant difference between Wx-B1b and Wx-D1b. The results of the double null lines further demonstrated that for the capacity of amylose synthesis, Wx-B1a was the highest, followed by Wx-D1a and Wx-A1a, and there was no significant difference between Wx-B1a and Wx-D1a. Amylose contents of the eight genotypes were not significantly correlated with plant height, spike length, spikelets per spike, grains per spike, 1000-grain weight, which showed that starch quality breeding could integrate with high yield breeding. Amylose contents of the eight genotypes were negatively significantly correlated with SDS-sedimentation value(r=-0.726), which suggested that reduction in amylose content is propitious to improve quality at a certain extent. Starch pasting properties of the full waxy type was significantly different from the other seven types, with the highest peak viscosity and breakdown, and lowest valley viscosity, final viscosity, setback, peak time, pasting temperature and starting pasting temperature. It indicated that waxy wheat starch has special use in food and industry. Breakdown was negatively significantly correlated with amylose contents (r=-0.969), and the other parameters were positively significantly correlated with amylose contents (r=0.797 for final viscosity, r=0.910 for trough viscosity, r =0.954 for setback, r =0.970 for peak time, r=0.962 for pasting temperature and r=0.932 for starting pasting temperature ). The results of the study are very useful for waxy wheat variety breeding and starch quality improvement.
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课题组在不断地创制新的同源四倍体材料的同时,连续多年以提高结实率为目的培育、筛选自交系材料,已获得自交繁殖十二年的高代自交系材料。相对于诱导创制初期,材料表现出的结实率低,同种系单株间的差异较大;高代材料已表现出较显著的结实率提升和较一致的农艺性状表型。 本实验选取课题组多年培育的同源四倍体水稻高代自交系材料,通过形态学、农艺性状和细胞遗传学比较,研究水稻同源四倍体与二倍体之间的异同。结果显示,所有同源四倍体材料的染色体组成均为2N=4X=48,花粉母细胞(PMC)减数分裂行为较正常,99%以上的染色体都能在减数分裂中期I(MI)发生联会、配对,形成四价体和二价体,这与理论染色体组构成相符。在减数分裂过程中,结实率较高的材料染色体异常现象较少而结实率较低的材料染色体异常现象较严重。统计分析表明,二价体和四价体的比例对结实率没有显著影响,但是三价体的数目对结实率有一定影响。这一结果表明了结实率和细胞减数分裂行为可能存在相关性,同源四倍体的减数分裂行为为筛选高结实率的同源四倍体种系提供了依据。 然后,对同源四倍体水稻高代自交系材料进行农艺性状和品质性状的统计与分析。主要针对结实率、每穗实粒数、有效分蘖和穗长等主要农艺性状,以及直链淀粉含量这一重要的品质性状进行统计。将统计结果与1996年诱导加倍的初代材料的数据相对比分析,结果显示所有材料经过多代选育培养,其农艺性状已经有了较显著的提高,同时同源四倍体材料的农艺性状稳定性也有了较显著的提升。如结实率的提高幅度较大,所有材料的平均结实率均显著高于加倍初代,而同种材料不同单株间的结实率差异也显著地减少,变异系数(CV)的平均值由1996年的41.15%减少到了2008年的28.81%。其他重要农艺性状也有不同程度的改良,种内变异系数也相应地降低。此外,实验测量了同源四倍体材料和来源二倍体材料的直链淀粉含量。分析结果显示,部分材料的直链淀粉含量与二倍体亲本产生了较显著的差异,这可能是诱导加倍过程中的遗传变异造成的;同源四倍体材料的种内变异系数(CV)平均值由1996年的6%下降到了2008年的3.88%,显示出在品质性状方面,同源四倍体材料的遗传稳定性也有较显著的增加。同源四倍体材料农艺性状经过多年的选育,表现出一定的提升,同时,经过多年自交纯化,所有材料种系内的性状差异逐渐缩小,说明同源四倍体水稻的遗传稳定性随着自交纯化而增强,这为同源四倍体水稻的进一步选育打下了良好的基础。 最后,通过测量连续两年的自交系材料的遗传多态性,分析材料间遗传差异和种群遗传结构,深入研究连续两代材料间的遗传差异,研究同源四倍体水稻与二倍体材料遗传稳定性之间的差异。实验采用18对SSR微卫星标记对连续两代15个材料,共94份样本进行差异分析。通过扩增条带长度多态性分析,计算不同材料以及同种材料不同世代间的遗传距离,构建同源四倍体和二倍体水稻的分子指纹库,并绘制聚类图。结果显示,同源四倍体和二倍体不同材料间的遗传差异比较大,遗传距离处于0.4757至0.2816之间;而相同品种不同世代材料间的遗传差异较小,但也表现出一定的遗传差异。同种同源四倍体材料不同世代间的遗传差异比二倍体材料更大,两代四倍体材料间遗传距离处于0.1359至0.0485之间;而两代二倍体材料间的遗传距离处于均小于0.0388。结果表明,同源四倍体水稻高代材料具有一定的遗传稳定性,但与来源二倍体材料相比,其世代间的遗传变异性仍然较强。这种结果说明,经过多代的自交纯化培育,同源四倍体水稻材料能够建立起相对稳定的遗传结构,同时,其强于二倍体亲本的变异性有能够为新品种的选育,农艺性状、品质性状的改良提供一定的遗传基础。此外,分析结果表明通过分子标记辅助检验,水稻材料间的遗传多态性能够有效地区分不同的品种,这为水稻品种的分子鉴定提供了一定的依据。 本研究从细胞学鉴定,农艺性状统计分析以及分子标记辅助聚类分析多方面地对同源四倍体水稻高代系进行了研究,对探究同源四倍体水稻的遗传规律,进一步揭示其遗传特性、农艺性状的遗传构成,为进一步选育优质的多倍体水稻提供了一定的理论依据。 This group insists on creating new Autotetraploid Rice (Oryza sativa L.) materials, while improving the seed-setting of them for many years, cultivated and selected the inbred line materials, has obtained the high generation inbred lines after twelve years cultivation. Compared to the early induced materials, which shown the low seed setting, and the large difference between the different plants in the same germ-line; the high generation materials have shown significant improvement in seed setting and more uniform phenotype agronomic traits. The autotetraploid rice high generation inbred lines material, which has been cultivated for more than 12 years, was chose in this experiment. The similarities and differences between autotetraploid and diploid rice was studied through morphological, agronomic and cytogenetic ways. The results showed that all the chromosome of autotetraploid materials are composed of 2N=4X=48, the pollen mother cells (PMC) meiosis behavior is normal, more than 99% chromosomes in metaphase I(MI) were federated and paired to form tetravalents or bivalents, which constitutes a consistent theory of genome. In the meiosis process, the material with a higher seed setting showed less chromosome abnormal than the material whose seed setting is lower. However, statistical analysis showed that the bivalent and tetravalent rate had no significant impact on seed setting, but the number of trivalent had a certain impact on seed setting. The result shows that the seed setting may be related to the meiosis behavior, which provides a basis to cultivate new autotetraploid germ line with high seed setting through the meiotic behavior. Furthermore, the agronomic and quality traits of autotetraploid rice high generation inbred material were statistically analyzed. The statistically analysis was focused on major agronomic traits such as: seed setting, grains per panicle, effective tillers and panicle length, as well as the important quality trait amylose content. The statistic data was compared with the data in 1996, when the first induced generation of autotetraploid material, and the result shows that after a multi-generation breeding, the agronomic traits has been significantly improved in all the materials, while the stability of agronomic traits also significant upgraded. For instant, the seed setting increased significantly, the average seed setting of all materials was significantly higher than the first induced generation, and the differences between different plants in the same species also significantly reduced, the average of the coefficient of variation (CV) was reduced from 41.15% in 1996 to 28.81% in 2008. Other important agronomic traits had improved in different degrees; the coefficient of variation within species is also reduced accordingly. In addition, the amylose content of autotetraploid and diploid materials was measured in this experiment. The results shows that the amylose content of some of the material differed from diploid parents significantly, it may caused by the genetic change during the inducing, autotetraploid materials intra-specific coefficient of variation (CV) average reduced from 6% in 1996 to 3.88% in 2008, shows that this is a significant increase of quality traits stability in autotetraploid rice. Agronomic traits of autotetraploid material shows some improvement after years of breeding, at the same time, after years of purification, all material within the germ-line gradually narrow the differences in traits indicates that autotetraploid rice genetic stability was enhanced, which laid a good foundation for the further autotetraploid rice breeding. Finally, this experiment studied the genetic differences between materials of two generations and researched the difference of genetic stability between diploid and autotetraploid rice materials through investigating the genetic polymorphism, genetic differences between materials and population genetic structure of inbred line materials of two consecutive years.18 pairs of SSR microsatellite markers for 15 materials of two generations were used in this experiment, and the total of 94 samples were analyzed. Through the amplification length polymorphism analysis of different materials and materials in different generations, the genetic distance between materials and generations was analyzed, a diploid and autotetraploid rice molecular fingerprint database and map rendering cluster were constructed. The result shows that the genetic distance is between 0.4757 to 0.2816 among different autotetraploid and diploid materials; the genetic distance between different generations of same species was less, but also shows a certain degree of genetic differences. The inter-generational genetic differences of autotetraploid materials were greater than of the diploid materials, which are 0.1359 to 0.0485 as the genetic distance; comparing with the 0.0388 of diploid materials. The result shows that high generation inbred autotetraploid rice material has a certain genetic stability, but the genetic variation between generations is still strong comparing with the source diploid materials. It indicates that, after many generations of purification cultivation, autotetraploid rice materials established a relatively stable genetic structure, at the same time, stronger variability than its diploid parents are useful in the breeding of new varieties, provides a genetic foundation to the agronomic and quality traits improvement. In addition, the analysis result shows that the through the molecular marker-assisted testing, rice genetic polymorphism between materials can effectively distinguish different species, provides a certain basis for molecular identification of varieties of rice. A series of investigation such as cytological identification, statistical analysis of agronomic traits, molecular marker-assisted cluster analysis was applied in this experiment to research genetic pattern of autotetraploid rice high generation inbred lines, revealed the genetic characteristics and the genetic composition of agronomic traits, provides a theoretical basis for the further selection of high quality autotetraploid rice.
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为探索碳离子束辐照对细胞中端粒酶活性的变化,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/μ12C6+),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)银染端粒重复序列扩增法检测不同剂量下细胞端粒酶活性的变化。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和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)。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。本实验使重离子在辐照治疗中的优势得以体现。
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The aim of this study is to investigate the effects of different doses of 100 MeV/u carbon ions on sweet sorghum seeds in order to improve crop yields and their sugar content. After irradiation, seeds were germinated and grown to 30 days, and others were sown in the field. At the end of harvesting season all planted seeds were picked separately and M2 generations obtained. The differences among the treatments were examined using the RAPID procedure. In the study done by using 38 primers; according to the amplification results, the differences among the various doses treatment were shown.
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利用X射线对体外培养的人肝癌细胞SMMC-7721和正常人肝细胞HL-7702进行辐照,以细胞克隆形成法测定细胞存活率,多聚酶链式反应银染端粒重复序列扩增法(Telomeric repeat amplification protocol,TRAP-PCR)检测细胞中端粒酶活性变化,探讨辐照后细胞存活与细胞中端粒酶活性变化的关系。结果表明,SMMC-7721和HL-7702细胞的存活率都随辐照剂量的增加而相应降低。在1-4Gy剂量范围内,两种细胞的端粒酶活性变化均呈剂量依赖性增强。但在1Gy、2Gy、3Gy剂量下,SMMC-7721其端粒酶活性较对照(0Gy)细胞低;当辐照剂量达4Gy时,端粒酶活性略高于对照细胞。在1-4Gy剂量范围内,HL-7702和SMMC-7721的细胞存活与端粒酶活性在变化趋势上呈负的关系。
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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
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We have developed a new experimental system based on a microfluidic chip to determine severe acute respiratory syndrome coronavirus (SARS-Cov). The system includes a laser-induced fluorescence microfluidic chip analyzer, a glass microchip for both polymerase chain reaction (PCR) and capillary electrophoresis, a chip thermal cycler based on dual Peltier thermoelectric elements, a reverse transcription-polymerase chain reaction (RT-PCR) SARS diagnostic kit, and a DNA electrophoretic sizing kit. The system allows efficient cDNA amplification of SARS-CoV followed by electrophoretic sizing and detection on the same chip. To enhance the reliability of RT-PCR on SARS-CoV detection, duplex PCR was developed on the microchip. The assay was carried out on a home-made microfluidic chip system. The positive and the negative control were cDNA fragments of SARS-CoV and parainfluenza virus, respectively. The test results showed that 17 positive samples were obtained among 18 samples of nasopharyngeal swabs from clinically diagnosed SARS patients. However, 12 positive results from the same 18 samples were obtained by the conventional RT-PCR with agarose gel electrophoresis detection. The SARS virus species can be analyzed with high positive rate and rapidity on the microfluidic chip system.
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The most biological diversity on this planet is probably harbored in soils. Understanding the diversity and function of the microbiological component of soil poses great challenges that are being overcome by the application of molecular biological approaches. This review covers one of many approaches being used: separation of polymerase chain reaction (PCR) amplicons using denaturing gradient gel electrophoresis (DGGE). Extraction of nucleic acids directly from soils allows the examination of a community without the limitation posed by cultivation. Polymerase chain reaction provides a means to increase the numbers of a target for its detection on gels. Using the rRNA genes as a target for PCR provides phylogenetic information on populations comprising communities. Fingerprints produced by this method have allowed spatial and temporal comparisons of soil communities within and between locations or among treatments. Numerous samples can be compared because of the rapid high throughput nature of this method. Scientists now have the means to begin addressing complex ecological questions about the spatial, temporal, and nutritional interactions faced by microbes in the soil environment.
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Temporal trends in total ozone for the St. Lawrence estuary were estimated from ground-based measurements at the NOAA/CMDL station in Caribou, Maine. Linear regression analysis showed that from 1979 to 1999 total ozone has decreased by about 3.3% per decade on an annual basis and ≤6.2% per decade on a monthly basis relative to unperturbed (pre-CFC) levels. The influence of increased ultraviolet-B (280–320 nm) radiation associated with ozone depletion on water column photochemical processes was evaluated by modeling the photobleaching of chromophoric dissolved organic material (CDOM). Linear regression analysis showed small (<0.5% per decade), but statistically significant upward trends in maximum noontime photobleaching rates. Most notably, positive trends in relative rates for May, June, and July, when maximum absolute rates are expected, were predicted. A global model based on TOMS ozone data revealed increases in photobleaching of ≤3% per decade at high latitudes in the Southern Hemisphere. Radiation amplification factors for increases in photochemically weighted UV (280–400 nm) in response to ozone depletion were estimated at 0.1 and 0.08 for photobleaching of CDOM absorbance at 300 and 350 nm, respectively. Application of the laboratory-based model to conditions that more closely resembled those in situ were variable with both overestimation and underestimation of measured rates. The differences between modeled rates and observed rates under quasi-natural conditions were as large or larger than the predicted increases due to ozone depletion. These comparisons suggest that biological activity and mixing play an important, but as yet ill-defined, role in modifying photochemical processes.
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The crystal structure of Er(PM)(3)(TP)(2) [PM = 1-Phenyl-3-methyl-4-isobutyryl-5-pyrazoloiie, TP = triphenyl phosphine oxide] was reported and its photoluminescence properties were studied by UV-vis absorption, excited, and emission spectra. The Judd-ofelt theory was introduced to calculate the radiative transition rate and the radiative decay time of 3.65 ms for the I-4(13/2) -> I-4(15/2) transition of Er3+ ion in this complex.
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Quantitative electrochemilumineseence (ECL) detection of a model protein, bovine serum albumin (BSA) was achieved via biotin-avidin interaction using an avidin-based sensor and a well-developed ECL system of tris(2,2'-bipyridine) ruthenium(II) derivative as label and tri-n-propylamine (TPA) as coreactant. To detect the protein, avidin was linked to the glassy carbon electrode through passive adsorptions and covalent interaction with carboxylate-terminated carbon nanotubes that was used as binder to immobilize avidin onto the electrode. Then, biotinylated BSA tagged with tris(2,2'-bipyridine) ruthenium(II) label was attached to the prepared avidin surface.