大肠杆菌基因组水平蛋白质-RNA相互作用初步研究

目的初步研究大肠杆菌中基因组水平的蛋白质-RNA相互作用（protein-RNA interactions,PRI）。方法通过RNA酶消化细菌裂解液,提取与蛋白质相互作用的RNA片段,构建cDNA文库,进行高通量测序,并通过生物信息学分析获得与蛋白质结合的转录本。结果获得了与蛋白质结合的3193条转录本,涉及2234个mRNA、47个sRNA（small regulatory RNAs）、39个tRNA、11个rRNA以及862个基因间区（intergenic region,IGR）。结论初步获得大肠杆菌中与蛋白质相互作用的转录本信息,为进一步开展PRI研究提供了支持。     

结合RNA的蛋白质位点预测研究

目的尝试通过构建数学模型来确定蛋白质中与RNA相互作用的氨基酸位点。方法从蛋白质结构数据库（protein data bank,PDB）中收集532例蛋白质与RNA相互作用的数据,对每个氨基酸提取150个特征,并利用机器学习方法构建2个与RNA结合的蛋白质位点预测模型。结果经过在同一数据集上与其他模型比较,所构建的模型具有更好的性能。结论该预测模型的建立,为研究人员获得与RNA结合的蛋白质位点提供了较好的生物信息学支持。     

鼠疫耶尔森菌菌株91001的蛋白质组学初步研究

目的建立鼠疫耶尔森菌的蛋白质组学研究方法,获得鼠疫耶尔森菌的基本蛋白质组数据。方法以对人无致病能力的布氏田鼠疫源地菌株91001为研究对象,按照溶解性的不同分别收集菌体蛋白,以3种不同方法(Shotgun-LC-MS-MS,1D-LC-MS-MS和2D-MS)进行分析,将分析数据与基于91001菌株基因组全序列建立的蛋白质理论数据库进行比对,确定91001菌株本实验培养条件下所表达的蛋白组分。结果Shotgun-LC-MS-MS方法鉴定了971种蛋白,1D-LC-MS-MS鉴定了915种,而2D-MS方法则鉴定了233种蛋白质,三者合计为1193种蛋白质,占基因组预测CDS的28．7％(1193／4143)。结论不同的蛋白质组分析方法鉴定的蛋白质种类和数目都存在差异,为更全面地获得鼠疫耶尔森菌蛋白质组数据,有必要同时采取多种方法进行分析。     

人血管生成素-1基因cDNA的克隆及结构分析

 目的 克隆人血管生成素-1基因,并分析其结构特征,为研究生理及病理性血管生成奠定基础。方法 提取人胎盘组织总RNA,用逆转录聚合酶链式反应(RT-PCR)技术扩增逆转录产物,经克隆及序列测定获得人血管生成素-1 cD-NA;用计算机软件分析基因序列及编码蛋白质结构。结果 获得高质量的胎盘组织总RNA。RT-PCR扩增出—1.5 kb的cD-NA片段,将PCR产物的阳性克隆测序,显示含1534 bp的插入片段,可编码503个氨基酸的蛋白质,与小鼠的血管生成素-1氨基酸高度同源。编码的氨基酸中含3个功能结构域。结论 为从基因及蛋白质水平研究人血管生成素-1与生理、病理性血管生成奠定了基础。     

细菌sRNA伴侣蛋白Hfq的研究进展

细菌sRNA能调节不同的功能,从结构调节到催化作用,影响生物体中各种各样的加工过程,包括RNA加工,mRNA稳定性,翻译,蛋白质稳定性和蛋白质分泌.小RNA(sRNA)通过多种机制发挥作用,其中许多需要RNA伴侣蛋白Hfq辅助.它们通过与靶RNA结合而参与到不同的RNA反应中.本文对sRNA伴侣蛋白Hfq的作用机制进行了综述.     

微量免疫比浊法测定血清免疫球蛋白、转铁蛋白和载脂蛋白含量

本研究建立了一种微量免疫比浊方法,可同时测定人体血清免疫球蛋白(IgG、IgA、IgM)、转铁蛋白(Tf)和载脂蛋白(Apo-AI,Apo-B).方法原理根据人体蛋白质分子的抗原特性,与抗体试剂发生特异性结合后形成微细颗粒复合物在PEG液中沉淀,经呈90度角的荧光照射后所产生的光散射强度求得相应的蛋白质含量.方法具有用血量微、灵敏度高和简单快速的特点.本文用此方法对我国优秀运动员的正常参考值进行了测定.     

精子蛋白质的合成及意义

Bhargara(1957)发现射出体外的牛精子,可以结合氨基酸形成蛋白质,并且蛋白质的合成与活力具有平行关系。但是没有发现RNA的存在,因此他错误地提到了精子细胞中蛋白质的合成是不需要RNA的观点,并得到一些研究者的承认。直到60年代许多研究者还认为精于内不含RNA。他于1959年还发现洗涤精子和非洗涤精子,结合     

CLIP相关技术的研究进展

在真核生物基因表达的转录后调节中,RNA结合蛋白（ RBP）起着关键作用,很多RBP的异常与人类疾病的发生密切相关。自2000年的RNA免疫沉淀和芯片分析方法（ RNA immunoprecipitation with differential display or microarray analysis , RIP-ChIP）出现以来,人们开始就RBP与RNA相互作用进行了系统而广泛的研究。经过改良和发展,基于体内实时紫外交联免疫沉淀法（ ultraviolet crosslinking and immunoprecipitation , CLIP ）、交联免疫沉淀cDNA文库高通量测序法（ high-throughput sequencing of CLIP cDNA library , HITS-CLIP）、光催化核糖核苷增强交联和免疫沉淀法（ photoactivatable-ribonucleoside-enhanced crosslinking and immunprecipitation , PAR-CLIP）以及提高个别核苷酸分辨率交联和免疫共沉淀法（ individual nucleotide resolution CLIP , iCLIP）等RIP-ChIP衍生方法相继产生,使用这些方法,可以解析RBP的RNA识别特异性,而且通过与高通量测序技术结合,可以实现转录组尺度的RBP的靶序列的鉴定,分辨率也得到极大提高。该文就RNA与蛋白的相互作用的基本原理及其研究进展、相关技术存在的问题以及发展趋势进行简要综述。     

氨基酸组成分析直接鉴定SDSPAGE分离的研究简报蛋白质

蛋白质组学是后基因组时代研究的重要领域。蛋白质组研究的基本技术包括双向凝胶电泳分离技术、蛋白质鉴定技术、计算机图像数据处理与蛋白质组数据库构建等。氨基酸组成分析[1,2 ] 作为蛋白质组研究中的鉴定方法之一 ,对质谱鉴定、Ｎ端氨基酸序列鉴定等具有补充及验证作用。蛋白质是由 2 0种氨基酸通过肽键连接起来的生物大分子 ,测定蛋白质的氨基酸组成可获得蛋白质的基本信息。本研究以马心肌红蛋白和细胞色素Ｃ为样品 ,优化实验条件 ,以此建立一种高灵敏度、高准确度的鉴定双向凝胶电泳分离后的蛋白质点方法。该方法还可用于分析天然蛋…     

δ因子在成人中的意义

δ因子是在急慢性乙型病毒性肝炎(HBV)病人中复制的RNA病毒.δ抗原外面为乙型肝炎表面抗原(HBsAg)所覆盖;并与病毒的RNA基因有关.1977年Mario Rizzetto首先报道在急慢性HBV病人的肝细胞核内发现了δ抗原.δ抗原可引起全身性免疫反应,首先是IgM应答;随后是IgG应答.将HBsAg自病毒颗粒分离后,就可以测出血清中的δ抗原以诊断某些急性感染.对这组病人,目前最好的诊断方法,用IgM型抗-δ免疫技术.对慢性感染者,用直接免疫荧光和免疫酶技术,可在固定和非固定的肝组织中找到δ抗原.此外,δ因子反复复制时,血清中常存在IgM抗δ,同时又与肝脏内δ因子密切相关.近代用cDNA探子来测定血清中的δ-RNA,证实了δ-RNA与δ因子的反复复制有密切关系,而且还证实肝脏内有δ抗原的存在.     

Knee injury and Osteoarthritis Outcome Score (KOOS) - validation of a Swedish version

The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living.     

Endovascular management of carotid-cavernous fistulas

Objective To investigate endovascular treatment of traumatic direct carotid-cavernous fistulas （CCF） and their complications such as pseudoaneurysms. Methods： Over a five-year period, 22 patients with traumatic direct CCFs were treated endovascularly in our institution. Thirteen patients were treated once with the result of CCF occluded, 8 twice and 1 three times. Treatment modalities included balloon occlusion of the CCF, sacrifice of the ipsilateral internal carotid artery with detachable balloon, coll embolization of the cavernous sinus and secondary pseudoaneurysms, and covered-stem management of the pseudoaneurysms. Results All the direct CCFs were successfully managed endovascularly. Four patients developed a pseudoaneurysm after the occlusion of the CCF with an incidence of pseudoaneurysm formation of 18.2% （4/22）. A total number of 8 patients experienced permanent occlusion of the ICA with a rate of ICA occlusion reaching 36.4% （8/22）. Followed up through telephone consultation from 6 months to 5 years, all did well with no recurrence of CCF symptoms and signs. Conclusion Traumatic direct CCFs can be successfully managed with endovascular means. The pseudoaneurysms secondary to the occlusion of the CCFs can be occluded with stent-assisted coiling and implantation of covered stents.     

The acutely limping child

Acute limping may be the result of multiple pathologies in children. The differential diagnosis varies based on the age of the child. Irrespective of age, the initial imaging work-up includes AP and frog leg radiographs of the pelvis and ultrasound; MRI may sometimes be helpful. In children less than 3 years, infections and trauma are most frequent. MRI is the imaging modality of choice when osteomyelitis is clinically suspected. Between the ages of 3 and 10 years, transient synovitis of the hip and Legg-Calvé-Perthes disease are main considerations but infection, inflammation and focal bony lesions are also considered. In children over 10 years, slipped capital femoral epiphysis also is considered.     

Do Balance Board Training Programs Reduce the Risk of Ankle Sprains in Athletes?

Introduction Ankle sprains are the most common musculo-skeletal injury that occurs in athletes,particularly in sports that require jumping and landing on one foot such as soccer,and basketball(1-4).These injuries often result in significant time loss from participation,long-term disability,and have a major impact on health care costs and resources(5-8).     

High Intensity Interval Training:New Insights

KEY POINTS ·High-intensity interval training(HIT)is characterized by repeated sessions of relatively brief，intermittent exercise.often performed with an“a11 out”effort or at an intensity close to that which elicits peak oxygen uptake(i.e.，≥90%of VO2 peak).     

Developmental validation of the PowerPlex(®) ESI 16 and PowerPlex(®) ESI 17 Systems: STR multiplexes for the new European standard

In response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega^® developed a suite of four new DNA profiling kits. This paper describes the developmental validation study performed on the PowerPlex^® ESI 16 (European Standard Investigator 16) and the PowerPlex^® ESI 17 Systems. The PowerPlex^® ESI 16 System combines the 11 loci compatible with the UK National DNA Database^®, contained within the AmpFlSTR^® SGM Plus^® PCR Amplification Kit, with five additional loci: D2S441, D10S1248, D22S1045, D1S1656 and D12S391. The multiplex was designed to reduce the amplicon size of the loci found in the AmpFlSTR^® SGM Plus^® kit. This design facilitates increased robustness and amplification success for the loci used in the national DNA databases created in many countries, when analyzing degraded DNA samples. The PowerPlex^® ESI 17 System amplifies the same loci as the PowerPlex^® ESI 16 System, but with the addition of a primer pair for the SE33 locus. Tests were designed to address the developmental validation guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), and those of the DNA Advisory Board (DAB). Samples processed include DNA mixtures, PCR reactions spiked with inhibitors, a sensitivity series, and 306 United Kingdom donor samples to determine concordance with data generated with the AmpFlSTR^® SGM Plus^® kit. Allele frequencies from 242 white Caucasian samples collected in the United Kingdom are also presented. The PowerPlex^® ESI 16 and ESI 17 Systems are robust and sensitive tools, suitable for the analysis of forensic DNA samples. Full profiles were routinely observed with 62.5 pg of a fully heterozygous single source DNA template. This high level of sensitivity was found to impact on mixture analyses, where 54–86% of unique minor contributor alleles were routinely observed in a 1:19 mixture ratio. Improved sensitivity combined with the robustness afforded by smaller amplicons has substantially improved the quantity of data obtained from degraded samples, and the improved chemistry confers exceptional tolerance to high levels of laboratory prepared inhibitors.