nm23-H1基因与人早幼粒白血病细胞HL-60增殖的相关性分析

目的探讨nm23-H1基因的表达与白血病细胞HL-60增殖之间的关系。方法：以25ng／mL阿糖胞苷处理HL-60细胞，MTT法测定细胞生长抑制率，NBT还原比色法判断细胞分化状况，RT-PCR检测nm23-H1基因表达的变化；构建nm23-H1基因的真核表达质粒pEGFP-N1-nm23-H1，转染HL-60细胞，通过细胞生长曲线和血清依赖性实验检测nm23-H1基因的过表达对HL-60细胞生长的影响。结果：小剂量Ara-C对HL-60细胞的生长呈时间依赖性抑制，作用4d后细胞NBT还原能力增强且nm23-H1基因的表达下调；转染nm23-H1基因的HL-60细胞生长加快、血清依赖性下降。结论：Ara-C对HL-60细胞增殖的抑制作用与下调nm23-H1基因的表达有-定关系；nm23-H1基因在HL-60细胞中的过表达有促细胞增殖的作用，即增高了HL-60细胞的恶性程度。     

SAFB1蛋白的表达纯化及其对乳腺癌细胞抑制作用的验证

利用RT-PCR方法从7721细胞系中分离野生型SAFB1基因,将该基因分别克隆入PET28a和pcDNA3.1载体来构建原核表达及真核表达载体.重组质粒经IPTG诱导表达后亲和层析纯化,用Western印迹实验验证蛋白表达的正确性.将SAFB1基因转入MCF7中,以RT-PCR的方法检测蛋白的表达,检测其对XOR基因的调控作用及做生长曲线验证其对乳腺癌细胞抑制作用.结果证实了SAFB1对乳腺癌细胞生长抑制作用非常明显,且对XOR基因的表达有一定的抑制作用.     

PinX1基因真核表达载体的构建及其对乳腺癌MCF-7细胞增殖的抑制作用

探讨了PinX1 基因在乳腺癌MCF-7 细胞生长和细胞周期中的作用, 初步探讨了该基因用于乳腺癌临床治疗的可行性. 采用RT-PCR 技术从293-T 细胞中扩增PinX1 基因, 将其克隆入真核表达载体pEGFP-C1 中, 再将重组质粒转染MCF-7 细胞. 通过real-time PCR 检测PinX1 基因的mRNA 表达, 用MTT 法检测转染前后细胞生长曲线的变化, 用流式细胞仪检测转染目的基因后细胞生长周期的改变. 检测结果表明, PinX1 基因已经在转染后MCF-7 细胞的细胞核内稳定表达, 乳腺癌细胞生长明显减缓(P <0.05), 增殖变慢(P <0.05), 细胞生长阻滞于G0/G1 期, 说明PinX1 基因可抑制乳腺癌MCF-7 细胞的生长和增殖.     

人TPO基因转染肝癌细胞后的表达及其受体水平的变化

从hTPO工程菌株中提取的pcDNA3/TPO质粒用脂质体包裹后转染人肝癌细胞株HepG2,并以未转染的HepG2和转染pcDNA3野生型质粒的HepG2作为对照，研究hTPO和其受体c-mpl在三组细胞中的表达情况以及HepG2细胞的生长情况，发现（1）RT－PCR法检测结果表明pcDNA3/TPO基因在转染HepG2细胞后，mRNA能有效表达，但对c-mpl的表达基本无影响；（2）细胞的生长计数和MTT法检测结果说明hTPO的有效表达对人肝癌细胞株HepG2的生长、繁殖有一定的抑制作用，同时还发现脂质体导入及pcDNA3型质粒的转染能够促进HepG2的生长。     

人端粒酶催化亚单位(hTERT)特异性核酶对乳腺癌细胞MCF-7活性的作用及影响

目的 探讨端粒酶特异性核酶对乳腺癌细胞MCF-7端粒酶活性、增殖和凋亡的影响.方法 设计合成RZ基因,构建人端粒酶锤头状核酶真核表达质粒pcDNA3.1( )-hTERT-RZ,测序鉴定.提取MCF-7细胞总RNA,RT-PCR扩增靶基因的cDNA片段,插入载体pMD18-T并测序鉴定.将核酶重组体及hTERT载体体外转录,琼脂糖凝胶电泳检测Rz的切割活性.核酶载体转染乳腺癌细胞MCF-7,RT-PCR法检测核酶重组子转染效率及核酶转染细胞中hTERT mRNA的表达量,TRAP法测定细胞端粒酶活性,MTT法、流式细胞仪测定细胞增殖及凋亡.结果 测序表明peDNA3.1( )-hTERT-Rz和pMD18-T-hTERT构建成功.5%琼脂糖凝胶电泳表明体外转录出Rz和靶基因hTERT,Rz对靶基因hTERT具有切割活性.构建好的核酶真核表达载体转染乳腺癌MCF-7细胞.发现其潜伏期延长,对数生长期减缓,S期细胞比例降低,G1期细胞比例增高(P＜0.01).端粒酶活性检测发现端粒酶活性明显降低.结论 端粒酶催化亚单位特异性核酶对靶基因hTERT在体外和体内均有催化切割活性,该核酶在乳腺癌MCF-7细胞中抑制hTERT的表达和细胞增殖,为核酶应用于恶性肿瘤的基因治疗提供了实验依据和新型工具酶.     

应用SCID鼠筛选肝癌转移性亚克隆及M-H7402亚细胞系的建立

本研究应用人肝癌细胞SCID鼠转移模型，筛选和建立肝癌转移细胞系，旨在为肝癌转移研究提供实验材料．采用人肝癌细胞H7402，腋后背部皮下接种SCID鼠，接种后66天时，在荷瘤鼠赢弱时引颈处死，取其肺组织，进行原代细胞培养和传代培养，获得肝癌细胞H7402的亚细胞克隆，命名为M—H7402．然后，对M—H7402细胞进行了生物学鉴定，检测了细胞形态、染色体、细胞动力学、细胞周期、甲胎蛋白表达、癌基因表达和转移相关基因表达等指标．结果显示，M—H7402细胞的生长、增殖和形态等与亲本细胞H7402十分相近，其染色体形态仍为人类核型，众数维持在72～80之间，占75．0％．RT—PCR检测结果显示，M—H7402细胞甲胎蛋白表达为阳性．Western blot检测结果显示，与亲本细胞H—7402相比，癌基因C—myc表达水平较高，转移抑制基因nm23的表达水平明显下调．从肺组织中获得的肝癌转移细胞系M—H7402，在体外可连续传代培养，细胞形态不变．应用SCID鼠筛选获得的亚细胞克隆，与亲本细胞形成配对关系，可为肝癌细胞转移的研究提供新的实验材料。     

抑癌基因p27在肿瘤细胞MCF7中的表达

抑癌基因p27是一个细胞周期依赖性激酶抑制子CKI(CDK inhibitor),对细胞周期起着负调控的作用,将p27基因克隆到表达载体pcDNA,转染到肿瘤细胞MCF7中,筛选到稳定表达株．p27基因的过量表达确实对肿瘤细胞的生长产生了抑制作用,并且引发了部分肿瘤细胞的凋亡．     

钾通道阻断剂对人乳腺上皮细胞MOF10A增殖的影响

探讨电压门控钾离子通道在乳腺上皮细胞增殖过程中的作用．通过MTT法检测了钾通道阻断剂TEA、电压门控钾通道阻断剂4-AP对人乳腺上皮细胞MCF10A增殖的影响并与乳腺癌细胞MCF7作了比较,免疫印迹方法观察了电压门控钾通道Kv1．2、Kv1．5的表达．研究发现,两种钾离子通道阻断剂对人乳腺细胞和乳腺癌细胞增殖的影响均呈剂量依赖性关系．经5mmol／LTEA处理72h后,MCF10A细胞的生长抑制率为21．67％,而MCF7细胞的生长抑制率为41．36％;5mmol／L 4-AP处理72h后,MCF10A的生长抑制率为29．24％,而MCF7细胞的生长抑制率为40．24％．Kv1．2在MCF10A和MCF7中表达没有变化．而Kv1．5在MCF10A细胞中的表达明显高于MCF7细胞．提示电压门控钾离子通道在乳腺细胞的增殖中起重要作用,其中Kv1．5可能和乳腺细胞的转化密切相关．     

pEGFP-IFI16表达载体构建及其表达对Hep-2细胞增殖的影响

RT-PCR扩增的IFI16基因连接到pUCm-T载体并测序鉴定,经测序正确的IFI16基因再连接到pEGFP-C1载体构建pEGFP-IFI16重组质粒,重组质粒pEGFP-IFI16转染Hep-2细胞后用荧光显微镜及半定量RT-PCR分析其表达情况,并用流式细胞仪测定细胞生长曲线分析其表达对Hep-2细胞增殖的影响.结果显示pEGFP-IFI16重组质粒构建正确,转染Hep-2细胞后荧光显微镜下观察到绿色荧光信号,半定量RT-PCR结果显示转染后Hep-2细胞IFI16基因条带亮度明显升高,细胞生长曲线测定结果显示转染后Hep-2细胞从第二天起其增殖速度变慢、至第三天时其增殖速度明显慢于对照细胞的增殖速度.说明成功构建了能在Hep-2细胞中表达EGFP-IFI16融合蛋白的pEGFP-IFI16重组质粒,pEGFP-IFI16重组质粒体表达能抑制Hep-2细胞的增殖.     

Fractalkine基因真核表达质粒的构建及其在小鼠肝癌细胞中的表达

为克隆小鼠趋化因子Fractalkine(．FK)基因，构建真核表达质粒，并在小鼠肝癌细胞中表达，用以进行肿瘤的基因治疗，用RT-PCR法，从小鼠乳腺癌细胞D2F2扩增FK的cDNA，插入pCR2．1 TOPO载体，测序证实后，将其亚克隆至质粒pIRES中构建FK真核表达载体；用脂质体将重组质粒转染小鼠肝癌MM45 T．Li细胞，经G418筛选获得抗性细胞克隆，用RT-PCR和免疫化学方法鉴定转染细胞中FK基因的表达．结果表明：经限制性内切酶酶切图谱分析和DNA序列测定证实目的基因已插入重组质粒，RT-PCR和免疫化学方法证明转基因MM45T．Li细胞克隆中存在小鼠FK基因的表达。     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that     

On the Thermodynamic Limit for Responses to Static Electromagnetic Fields

For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the     

An affordable accurate evaluation of the binding conformations predicted by molecular docking studies in bio-system

Franck-Condon factors bridge the gap between theoretical modeling and experimental observations for molecular electronic spectroscopy and electron transfer.Under the displaced harmonic oscillator approximation,multidimensional Franck-Condon factors are decomposed into a product of many one-dimensional(1D)Franck-Condon(FC)factors,and each 1D-FC factor is associated with one Huang-Rhys factor that determines the leading contribution of     

Call for Papers

<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with