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。     

hsa-miR-5688在肝细胞癌中靶途径预测体系的构建与验证

目的 构建并预测在原发性肝细胞癌(肝癌)中hsa-miR-5688的靶途径及调控机制,并通过TCAG数据库和串联质量标签(TMT)标记LC-MS/MS蛋白质测序方法验证其结果,以期构建准确高效的微小RNA(miRNA,miR)靶途径预测方案.方法 通过五大线上数据库筛选hsa-miR-5688靶基因,对细胞定位、分子功能、信号通路、疾病、组织表达和基因互作进行富集分析,构建分析策略,预测其靶途径及调控机制.利用TCAG数据库分析调控途径中靶基因的mRNA表达量、突变频率和生存曲线;基于肝癌复发和预后良好的肝癌组织标本,应用TMT标记蛋白质结合LC-MS/MS方法对靶途径中的互作蛋白进行定量检测,以验证预测体系的正确性.结果 预测获得hsa-miR-5688调控的靶基因,包括信使RNA(mRNA)3031个,环状RNA(circRNA)1511个;代谢通路、细胞定位、分子功能、疾病、组织表达等富集结果经多重分析发现,靶基因RHOBTB1和UBE2W共同参与的泛素化蛋白降解途径是其调控肝癌进程的最关键信号通路.TCAG临床数据分析表明,泛素化途径的靶基因mRNA表达量在肝癌组织中明显高于正常肝组织,且基因突变会明显降低患者生存率.TMT标记蛋白质结合LC-MS/MS分析表明,与RHOBTB1和UBE2W互作的蛋白质泛素化降解相关基因UBA5、NEDD4L、UBE3C和USP14直接参与肝癌复发调控.结论 hsa-miR-5688可通过泛素化蛋白降解途径参与肝癌发生和复发进程,证明通过优化生物信息学构建快速预测miRNA靶途径的体系具有一定的高效性和准确性.     

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。     

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.     

The role of interventional radiology in trauma

Introduction Interventional Radiology has evolved into a specialty having enormous input into the care of the traumatized patient.In all hospitals,regardless of size,the Interventional Radiologist must consider their relationships with the trauma service in order to     

Sexual homicide: Brief review of the literature and case report involving rape,genital mutilation and human arson

The objective of the current paper is to report a new case of sexual murder involving human arson and summarize the literature on the phenomenon of sexual homicide. The present case study is unprecedented in Greece and a rarity in international literature due to the fact that the victim suffered genital mutilation and incineration while still alive. The evaluation consisted of 176 articles; 53 were reviewed by the authors. The results revealed sparse, but significant, research findings. The authors discuss the limitations regarding research, incidence of the phenomenon, crime-scene patterns, offender characteristics (killing methods, motive inferences, sociodemographic data, classifications, psychopathology, modus operandi), and victim selection. The incidence of the phenomenon is unclear (1–4%) due to non-standardized criteria. It is an expression of displaced anger or sexual sadism and/or a way to elude detection (ancillary benefit). Most offenders (in their first kill) and victims were in their late 20s to early 30s and belong to Caucasian populations. Personal weapons were commonly used against women, strangulation is the prevalent killing method against children, and firearms against men. Most of the sexual homicide perpetrators are non-psychotic at the time of the attack, but experience personality pathology, primitive defenses, pathological object relations, and withdrawal into fantasy in order to deal with social isolation.     

Extension du modèle puissance–temps limite pour estimer la production d’énergie aérobie et anaérobie lors de l’exercice intense

Aim

Examine how the modelling of the relation between power and time to exhaustion can provide an estimation of the production of aerobic and anaerobic energy during intense exercise.

Current knowledge

The hyperbolic model made it possible to define the critical power corresponding to the maximal rate of energy renewed by aerobic metabolism. A new model distinguishing the critical power from the maximal aerobic power has been built to estimate more precisely the anaerobic contribution. Data from middle distance runners and subjects tested on cycle ergometer showed a relative contribution of anaerobic metabolism arising from critical power and increasing until around 10 % of total power when aerobic energy production reaches its maximum.

Prospects

Considering the slow component of oxygen uptake would provide a more precise analysis of energy production and transformation during exercise at high intensity.     

Beziehungen zwischen Aufprallgeschwindigkeit,Fahrzeugbeschädigungen,Frakturen und „Wurfweite” bei 50 tödlichen Fußgänger-PKW-Unfällen

Zusammenfassung Aus 160 Obduktionen von Fußgängern, die durch PKW getötet worden waren, sind 50 nach folgenden Gesichtspunkten ausgewählt worden:Auffahrunfälle Erwachsener mit gesicherten Angaben in den Gerichtsakten über Fahrzeugbeschädigungen, Zusammenstoßstellen, Endlage der Fußgänger und Bremsspuren. Aus diesen Daten wurden Ausgangs- und Aufprallgeschwindigkeiten berechnet sowie die Wurfweite der Fußgänger gemessen.Die Ausgangsgeschwindigkeiten, nach der Berechnung zwischen 32 und 95 km/h, lagen meistens etwas höher als die angegebenen Geschwindigkeiten. Zwischen Ausgangs- und Aufprallgeschwindigkeiten waren die Differenzen größer. Ab 12 km/h Aufprallgeschwindigkeit kam es bereits zu tödlichen Verletzungen. Auffallend ist die zweigipfelige Verteilung der Häufigkeit sowohl bei der Einteilung nach den Aufprallgeschwindigkeiten als auch nach den Deformationsarbeiten (Aufprallgeschwindigkeit: 26% bei 51–60 km/h und 22% bei 21 bis 30 km/h; Deformationsarbeit: 34% bei 39–200 kpm und 30% bei 701–1000 kpm).Bei Aufprallgeschwindigkeiten über 50 km/h wurden doppelt so viel Knochenbrüche an der Wirbelsäule und am Becken als bei Aufprallgeschwindigkeiten unter 50 km/h festgestellt.Bei 44 Fußgängerunfällen lag ein Drittel innerhalb der Erwartungsgrenze der Wurfweite nach Fiala, je ein Drittel aber darüber bzw, darunter.Das Beschädigungsbild und die Anstoßverletzungen geben Hinweise zur Ermittlung der Aufprallgeschwindigkeit, vor allem wenn weitere Berechnungsgrundlagen fehlen.

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Summary Fifty cases were selected from 160 autopsies performed on pedestrians who were accidentally killed by private motorcars; the cases were selected according to the following criteria:The cases were confined to impact accidents involving adults in which reliable evidence could be gathered from the Court records in relation to damage to the car, the place of collision, the final position of the injured pedestrian and the extent of skidmarks. The initial and collision speeds of the cars were calculated from these facts and the distance of projection of the pedestrians were measured.The initial speeds, calculated between 32 and 95 km/h, were, in most cases, higher than the declared speeds. The differences between initial and collision speeds were larger. Fatal injuries could arise from a collision velocity of only 12 km/h. A striking feature of the analysis was the distribution of two peaks of frequency whether these were classified according to the collision speeds or according to the deformation of the vehicle (collision speed: 26% at the speed of 51 to 60 km/h and 22% at the speed of 21 to 30 km/h; deformation work: 34% at the deformation work of 39 to 200 kpm and 30% at the deformation work of 701 to 1,000 kpm).Twice as many fractures of the vertebral column and pelvis were sustained at collision speeds over 50 km/h as were sustained under 50 km/h.In one third of 44 pedestrian accidents, the distance of projection was within the expected range described by Fiala; in one third the distance was above and in one third below the expected range.If additional data for calculation were lacking, the patterns in injury arising from the impacts gave indications from which it was possible to estimate the collision speed.

Stipendiat der Alexander v. Humboldt-Stiftung, Dozent Dr. med. S. Kamiyama, Dept. of Legal Medicine, School of Medicine, Chiba University, 313 Inohanacho, Chiba, Japan.     

Diagnóstico ecográfico del neumotórax

The ultrasonographic diagnosis of pneumothorax is based on the analysis of artifacts. It is possible to confirm or rule out pneumothorax by combining the following signs: lung sliding, the A and B lines, and the lung point. One fundamental advantage of lung ultrasonography is its easy access in any critical situation, especially in patients in the intensive care unit. For this reason, chest ultrasonography can be used as an alternative to plain-film X-rays and computed tomography in critical patients and in patients with normal plain films in whom pneumothorax is strongly suspected, as well as to evaluate the extent of the pneumothorax and monitor its evolution.     

Carbohydrate Supplementation during Exercise: Does It Help? How Much is Too Much?

KEY POINTS· Carbohydrate intake during exercise can delay the onset of fatigue and improve performance of prolonged exercise as well as exercise of shorter duration and greater intensity (e.g., continuous exercise lasting about 1h and intermittent high-intensity exercise), but the mechanisms by which performance is improved are different.     

磁共振DWI和动态增强鉴别颅底脊索瘤和侵袭性垂体瘤的应用

目的 探讨磁共振扩散加权成像(DWI)和动态增强在颅底脊索瘤和侵袭性垂体瘤(IPA)鉴别诊断中的应用价值.方法 搜集经手术病理证实且影像学有鞍区破坏的颅底脊索瘤患者15例、向鼻咽部侵犯的IPA患者20例.测量二者的表观扩散系数(ADC)值,绘制受试者工作特征曲线(ROC),分析动态增强曲线的类型,统计达峰时间(TTP)、增强峰值(EP)和最大对比增强率(MCER),分析各个参数在鉴别诊断中的价值.结果 颅底脊索瘤的ADC值为(1.274±0.07)×10-3mm2/s,高于IPA ADC值(0.672±0.03) ×10-3 mm2/s(P ＜0.001),ADC阈值为0.964×10-3mm2/s时,ROC曲线下面积为0.997,敏感度为93.3％,特异度为100％.颅底脊索瘤时间-信号强度曲线(TIC)Ⅰ型14例,TICⅢ型1例,此例TICⅢ型者TTP约40 s;IPA TIC Ⅰ型7例,TICⅢ型13例.颅底脊索瘤和IPA的EP、MCER差异均有统计学意义(P ＜0.001).结论 ADC值和TIC的类型及其相关参数(EP,MCER)有助于颅底脊索瘤和IPA之间的鉴别.     

Hip and pelvic injuries in the elderly

Fractures of the hip and pelvis are frequent and serious injuries in elderly patients. Due to the aging population, their incidence should double by 2050. Therefore, the social and economical implications of these fractures are significant. Delay in diagnosis increases the associated morbidity and mortality. The purpose is to review the imaging features of these fractures, the imaging techniques (projections, CT) to depict them and their classification based on severity.