手足口病患儿柯萨奇病毒A10型山东地方株VP1区基因特征分析

目的 了解手足口病(HFMD)患儿标本中分离到的柯萨奇病毒A10型(Coxsackie virus A10,CVA10)山东地方株VP1区基因特征.方法 采集2008年至2009年部分HFMD病例临床标本,细胞培养方法分离病毒,提取病毒核酸,RT-PCR扩增病毒VP1区序列,生物信息学方法鉴定其基因型别.选择代表性CVA10毒株,与GenBank中检索到的其他CVA10毒株进行核苷酸和氨基酸同源性分析,并以VP1部分序列构建亲缘进化树.结果 从山东省HFMD患儿的760份临床标本中分离出330株病毒,其中有17株分子定型鉴定为CVA10.CVA10山东地方株彼此间核苷酸和氨基酸的同源性分别在82.3%～100.0%和94.2%～100.0%,与CVA10原型株(Kowalik株)核苷酸和氨基酸的同源性为75.6%～76.8%和90.2%～93.2%.亲缘进化树显示,CVA10山东地方株分别属于两个不同的基因亚型.结论 CVA10也是引起HFMD的病原体之一,山东省存在CVA10两种基因亚型的共循环.     

2011年上海地区轻、重症手足口病患者肠道病毒71型分离株VP1、VP4区的基因特征分析

目的 了解2011年上海地区手足口病重症和轻症患儿中肠道病毒71型(EV71)分离株VP1、VP4区的基因特征.方法 对来自2011年重症与轻症手足口病患儿的各5株EV71分离株进行VP1、VP4全序列的RT-PCR扩增测序,并与美国国立生物技术信息中心公布的EV71 A、B、C基因型代表株进行核苷酸、氨基酸比对分析和系统进化分析.结果 轻、重症患儿的EV71分离株之间VP1基因的核苷酸同源性为96.0％～98.1％;VP4基因的核苷酸同源性为93.7％～99.5％.轻、重症患儿的EV71分离株与C基因型代表株比较接近,VP1区核苷酸同源性分别为86.9％～98.2％、87.4％o～98.5％,VP4区核苷酸同源性分别为85.5％～100.0％、84.5％～99.5％,其中与2008年安徽省阜阳市的EV71流行株(C4亚型)VP1区核苷酸同源性分别可达97.0％～98.2％、97.9％～98.5％,VP4区核苷酸同源性分别可达96.1％～100.0％、97.1％～99.5％.3例重症患儿分离株在VP1和VP4的天冬酰胺(N)282丝氨酸(S)、苏氨酸(T)7丙氨酸(A)同时发生变异.结论 2011年上海地区10例轻、重症手足口病患儿中分离的EV71流行株均属C基因型的C4亚型;3例重症患儿分离株在VP1和VP4的N282S、T7A同时发生变异.     

2016-2018河北地区儿童病毒性脑炎流行特征调查

目的了解河北地区儿童病毒性脑炎病原谱和分子进化特征。方法收集2016-2018年河北地区351例诊断为病毒性脑炎的患儿的临床资料。采用实时荧光定量PCR检测脑脊液中病原,并利用PCR扩增肠道病毒VP1基因序列片段。对柯萨奇病毒B5型测序并利用软件MEGA6.0绘制种系发生树,采用邻位相连法构建进化树。结果351例患者中,学龄前儿童247例占70.37%。其中发热312例、287例头痛、228例呕吐或腹泻、61例咽痛或有呼吸道症状、23例嗜睡、皮疹9例和惊厥3例。351份脑脊液样品中102份样品检测结果为阳性,阳性率29.06%。其中单一感染101例,双重感染1例。102份样本中,检出EVs 72份,VZV 15份,HSV 10份,检出HHV 6 3份,EBV与EV合并感染检出1份。72份肠道病毒样本中全部为肠道病毒B群,未检出肠道病毒A群。31株CVB5、18株Echovirus 30、3株CVB3、3株Echovirus 9、2株CVB2、1株CVB1。31株CVB5与其他中国株之间VP1核苷酸同源性90.1%～99.5%,氨基酸同源性92.8%～99.7%;18株Echovirus 30与其他中国株之间VP1核苷酸同源性91.6%～98.7%,氨基酸同源性95.3%～99.2%。系统进化分析显示CVB5均在C分枝上。结论发热和头痛是病毒性脑炎患儿的主要临床特征,肠道病毒是主要病原体。CVB5与其他中国株在同一分枝,说明亲缘关系较近。     

2016-2018河北地区儿童病毒性脑炎流行特征调查北大核心CSCD

目的了解河北地区儿童病毒性脑炎病原谱和分子进化特征。方法收集2016-2018年河北地区351例诊断为病毒性脑炎的患儿的临床资料。采用实时荧光定量PCR检测脑脊液中病原,并利用PCR扩增肠道病毒VP1基因序列片段。对柯萨奇病毒B5型测序并利用软件MEGA6.0绘制种系发生树,采用邻位相连法构建进化树。结果351例患者中,学龄前儿童247例占70.37%。其中发热312例、287例头痛.228例呕吐或腹泻.61例咽痛或有呼吸道症状.23例嗜睡、皮疹9例和惊厥3例。351份脑脊液样品中102份样品检测结果为阳性,阳性率29.06%。其中单--感染101例,双重感染1例。102份样本中,检出EVs 72份,VZV 15份,HSV 10份,检出HHV63份,EBV与EV合并感染检出1份。72份肠道病毒样本中全部为肠道病毒B群,未检出肠道病毒A群。31株CVB5、18株Echovirus 30、3株CVB3.3株Echovirus9、2株CVB2.1株CVB1。31株CVB5与其他中国株之间VP1核苷酸同源性90.1%~99.5%,氨基酸同源性92.8%~99.7%;18株Echovirus30与其他中国株之间VP1核苷酸同源性91.6%~98.7%,氨基酸同源性95.3%~99.2%。系统进化分析显示CVB5均在C分枝上。结论发热和头痛是病毒性脑炎患儿的主要临床特征,肠道病毒是主要病原体。CVB5与其他中国株在同一分枝,说明亲缘关系较近。     

2010年福建省肠道病毒71型分离株的基因型特征研究

摘 要:目的 全面研究和了解肠道病毒71型在福建省的遗传背景,分析和探讨肠道病毒71型在我省的基因型地理分布特征及传播特征。方法 对2010年福建省9个设区市的 50株EV71分离株进行VP1区全长基因序列测定,并对核苷酸序列进行同源性比较及遗传进化分析。结果 2010年福建省50株EV71型分离株的VP1区核苷酸序列全长均为891个bp,核苷酸及氨基酸序列同源性比较,50株EV71型分离株之间的VP1区核苷酸序列同源性为95.4%-100%,编码蛋白氨基酸序列同源性为97.3%-100%,与08年阜阳流行株Fuyang17.08-2同源性最高,核苷酸序列同源性为97.1%-99.3%,氨基酸序列同源性为98.7%-100%;VP1区基因遗传进化分析,与Fuyang17.08-2株进化关系较为接近,同属于C4a基因亚型。结论 2010年福建省50株EV71型分离株均属于C4a基因亚型,未产生明显的抗原漂移及变异;福建省及各个地市均分布亲缘关系较近的C4a基因亚型的EV71多传播链病毒,应加强长期动态地监测和分析。关键词:肠道病毒71型;VP1区;基因型特征;     

肠道病毒71型VP1蛋白二级结构及B细胞表位的预测

目的预测C4基因亚型肠道病毒71型的VP1蛋白二级结构及B细胞表位,为EV71疫苗研制提供理论基础。方法本研究将宁波地区EV71分离株与国内外EV71代表株进行同源性分析和并构建亲缘进化树。以VP1基因序列为材料,应用EX-PASY服务器上的GOR4、SOPMA两种方法分析预测蛋白质二级结构,并结合蛋白质的亲水性、柔韧性、表面可能性等指标综合评价VP1蛋白的B细胞抗原表位。结果同源性分析和亲缘进化分析得出,2010年宁波EV71分离株属于C4a基因亚型,与基因库检索到的C4a基因亚型代表株核苷酸和氨基酸同源性较高,分别为93.6%～99.3%和98.3%～100%。VP1蛋白二级结构以无规则卷曲和β-片层为主,有多处抗原指数较高的区段,结合亲水性、柔韧性、表面可能性等指标,综合预测VP1蛋白的B细胞抗原表位在第39～40、159～167、212～220、287～290位氨基酸残基的可能性较大。结论 EV71VP1蛋白多个区段具有抗原潜力通过生物信息学方法预测VP1蛋白二级结构及B细胞表位为EV71疫苗研制提供理论基础。     

新疆生产建设兵团手足口病柯萨奇病毒A6型的流行及其基因特征分析

目的了解新疆生产建设兵团地区柯萨奇病毒A6(CVA6)的流行特点并对其基因特征进行分析。方法对2014-2016年新疆生产建设兵团地区收集的手足口病非肠道病毒71型(EV71)非柯萨奇病毒A16型(CVA16)阳性标本进行CVA6荧光定量PCR检测,采用RT-PCR扩增CVA6阳性标本中VP1编码区基因,并进行核苷酸序列测定与分析;利用MEGA6.06与GenBank代表株进行核苷酸同源性分析并构建系统进化树。结果2014-2016年采集的CVA6阳性标占在非EV71非CVA16阳性标本的82.39%,68株CVA6流行株之间VP1区核苷酸和氨基酸序列同源性分别为92.5%~100.0%和98.9%~100.0%,系统发育树显示97条CVA6序列共形成A-D 4个分支,其中D分支进一步分为D1-D3 3个亚分支。CVA6流行株中的5株处于D2亚分支,63株处于D3亚分支。结论新疆生产建设兵团地区2014-2016年手足口病非EV71非CVA16肠道病毒以CVA6为主,基于VP1系统发育分析分为A-D 4个分支,D分支中的D2和D3亚分支并存,其中D3亚分支是优势流行株。     

北京市西城区手足口病柯萨奇病毒A5型VP4区基因特征分析

目的分析2009年北京市西城区手足口病柯萨奇病毒A5型(CoxA5)的VP4区基因特征,了解其变异情况。方法采集手足口病患儿咽试子,提取病毒RNA,用肠道病毒VP4区分型引物进行半巢式RT-PCR(RT-nPCR),通过测序和GenBank Blast比对确定肠道病毒型别,并对其VP4区进行序列和进化分析。结果共鉴定出5株CoxA5,GenBank登录号为JN981017-JN981021。在VP4区,5株CoxA5与中国株HQ728261核苷酸及氨基酸序列同源性分别为96%～99%和98%～100%。进化分析表明,5株CoxA5与中国株HQ728261遗传距离为0.005～0.045,共同构成一个独立的进化树分支。5株CoxA5在VP4区无氨基酸的缺失和插入,未发现特有的氨基酸置换。结论 2009年北京市西城区手足口病病原体CoxA5VP4区与中国株HQ728261在进化上密切相关,其序列未发生明显变异。     

2008-2009年江苏省肠道病毒71型分子流行病学研究

目的探讨2008-2009年江苏省肠道病毒71型(EV71)分离株的病毒基因特征。方法选取18份EV71分离株,用一对特异引物进行VP1全基因核苷酸序列扩增,在ABI3130型自动测序仪上测序,序列结果用DNA Star和MEGA4软件进行分析,并与各型参考株进行比较。结果 18株病毒与安徽省阜阳市和山东省临沂市分离的C4a基因型参考株最为相近,核苷酸同源性在96.3%~99.6%之间,氨基酸同源性在99.0%~99.7%之间;而与A、B基因型参考株比较差异较大,核苷酸同源性只在81.8%~84.4%之间,氨基酸同源性在94.3%~97.0%之间;说明本文的18株病毒均属于EV71型C4a基因型。结论 2008-2009年江苏省分离的18株EV71可能与安徽省和山东省分离的EV71有相同的起源,均属于C4a亚型;目前C4a亚型病毒在中国大陆可能有较广泛的分布和传播。     

手足口病柯萨奇病毒A10和A6型的分子鉴定及其VP1区3′端序列分析

目的鉴定手足口病患儿咽拭子病原体柯萨奇病毒A10(CVA10)和A6(CVA6),并对其VP1编码区3′端序列进行分析。方法合成对肠道病毒VP1区3′端序列具有较高特异性的兼并引物040-011,RT-PCR方法测定其VP1区3′端基因序列,通过BLAST程序比对,鉴定病毒的基因型;与GenBank中其他已知病毒基因型序列进行同源性分析,构建遗传进化树。结果 38例非EV71非CVA16引起的手足口病患儿咽拭子,5例检测到CVA10,1例检测到CVA6。在进化关系上CVA10与D基因型最为密切,核苷酸和氨基酸同源性分别为92.7%~94.2%、91.7%~100%;CVA6与2003~2009年报告的部分毒株核苷酸同源性为86.8%~90.2%,其中与2008年台湾EU908148的毒株同源性最高;氨基酸同源性为85.2%~89.0%。结论兼并引物040-011对CVA10和CVA6有良好的基因扩增特异性;引起手足口病的肠道病毒除EV71和CVA16外,CVA10和CVA6也是重要的病原;鉴于2008年芬兰和2009年山东省文登市CVA10引起的手足口病暴发,应加强对手足口病病原的检测和分型,以了解柯萨奇病毒...     

Use of fab fragments of digoxin-specific antibodies in the therapy of massive digoxin poisoning

A case of massive digoxin ingestion with multiple arrhythmias, consisting of high grade A-V block and ventricular ectopy not responsive to lidocaine, is described. The arrhythmias ceased following administration of digoxin-specific Fab fragments. The patient improved and was transferred to the psychiatric unit.     

Control of intraabdominal hemorrhage and shock: a comparison of fluid resuscitation, MAST, and balloon occlusion

Our study examined the efficacy of four treatment modalities in controlling hemorrhage and achieving hemodynamic stabilization in hemorrhagic shock: intravenous fluid replacement (IV); military antishock trousers used concomitantly with fluids (MAST); balloon occlusion at the level of the diaphragm with concomitant fluid replacement (balloon); and a combination of MAST inflation, balloon occlusion, and fluid resuscitation (MAST and balloon). Twenty-eight mongrel dogs were anesthetized, and the spleen was exposed and completely crushed. The abdomen was closed, and treatment was initiated and continued for four hours or until the dog died. For all conditions the hematocrit dropped during the course of the experiment; balloon occlusion was effective at slowing this drop (P less than .0001), but MAST had no statistically significant effect. Animals with balloons bled more slowly into the abdominal cavity than did animals in the other two groups (P less than .0001). MAST also were effective at slowing the bleeding (P less than .05). Of the balloon and the MAST and balloon dogs, all except one survived the entire four hours; this difference between balloon and nonballoon dogs is significant (P = .002). MAST did not have a statistically significant effect on survival. Perfusion pressure (PP) declined during the course of the experiment, and the balloon was effective at slowing this decline (P less than .0001); none of the other comparisons was statistically significant.     

Comparison of intravenous and oral routes of theophylline loading in acute asthma

In a prospective, randomized clinical trial, 19 patients with an acute exacerbation of asthma were given a loading dose of aminophylline by the IV (n = 10) or oral route (n = 9) of administration following treatment with epinephrine. Plasma concentrations of theophylline were measured prior to giving the loading dose, and one, two, three, and 24 to 48 hours later. Therapeutic effectiveness was evaluated by analyzing spirometric measurements prior to giving the loading dose, and one, three, and 24 to 48 hours later. Side effects also were recorded. In the IV group, the mean peak plasma theophylline concentration was 15.1 micrograms/mL one hour after loading, and in the oral group the mean peak serum theophylline concentration was 14.2 micrograms/mL three hours after loading. There was no correlation between theophylline concentrations and normalized change in spirometric values. There was no significant difference in spirometric values between the IV and oral groups. Nausea was slightly more common in the IV group. We conclude that there is no therapeutic advantage to giving a loading dose of aminophylline by the IV route rather than orally in patients with mild-to-moderate exacerbation of asthma initially treated with epinephrine.     

Role of microRNA in regulation of myeloma-related angiogenesis and survival

Multiple myeloma (MM) is a malignant disease caused by clonal proliferation of plasma cells that result in monoclonal gammopathy and severe end organ damage. Despite the uniform clinical signs, the disease is very diverse in terms of the nature and sequence of the underlying molecular events. Multiple cellular processes are involved in helping the malignant cells to remain viable and maintain proliferative properties in the hypoxic microenvironment of the bone marrow. Specifically, the process of angiogenesis, triggered by the interactions between the malignant MM cells and the stroma cells around them, was found to be critical for MM progression. In this review we highlight the current understanding about the epigenetic regulation of the proliferation and apoptosis of MM cells and its dependency on angiogenesis in the bone marrow that is carried out by different microRNAs.     

Application of Granular Biocomposites Based on Homogenised Peat for Absorption of Oil Products

Among the various methods for collecting oil spills and oil products, including from the water surface, one of the most effective is the use of sorbents. In this work, three-component bio-based composite granular adsorbents were produced and studied for oil products’ pollution collection. A bio-based binder made of peat, devulcanised crumb rubber from used tyres, and part fly ash as cenospheres were used for absorbent production. The structure, surface morphology, porosity, mechanical properties, and sorption kinetics of the obtained samples were studied. Composite hydrophobicity and sorption capacity to oil products, such as diesel fuel (DF) and motor oil (MO), were determined. The obtained pellets are characterised by a sufficiently pronounced ability to absorb oil products such as DF. As the amount of CR in the granules increases, the diesel absorption capacity increases significantly. The case of 30-70-0 is almost three times higher than the granules from homogenised peat. The increase in q is due to two factors: the pronounced surface hydrophobicity of the samples (Θ = 152°) and a heterogeneous porous granule structure. The presence of the cenosphere in the biocomposite reduces its surface hydrophobicity while increasing the diesel absorption capacity. Relatively rapid realisation of the maximum saturation by the MO was noted. In common, the designed absorbent shows up to 0.7 g·g⁻¹ sorption capacity for MO and up to 1.55 g·g⁻¹ sorption capacity for diesel. A possible mechanism of DF absorption and the limiting stages of the process approximated for different kinetic models are discussed. The Weber–Morris diffusion model is used to primarily distinguish the limiting effect of the external and internal diffusion of the adsorbate on the absorption process.     

RNA-Seq在老年神经退行性疾病研究中的应用

阿尔茨海默病及帕金森病是老年人最常见的两种神经退行性疾病,但其发病机制及治疗是研究的热点。随着高通量测序技术的进步及成本的下降,RNA-Seq也成为神经退行性疾病机制研究及生物标志物发现的有力手段。RNA-Seq相对于microarray具有高灵敏度、高准确性、高重复性以及噪声低等优势,在阿尔茨海默病及帕金森病研究中有较为广泛的应用,包括检测差异表达基因,可变剪接、新长链非编码RNA预测分析和miRNAs调控等,但是容易受病理复杂性及样本等因素影响。目前阿尔茨海默病及帕金森病转录组研究相比于癌症等还不够深入,在临床诊断及治疗应用还面临较大挑战。但是随着新技术及新方法的发展,RNA-Seq将进一步推动神经退行性相关疾病的研究和临床转化。     

岳阳市城区小学生血吸虫病防治健康教育现状调查

目的 调查湖南省岳阳市城区小学生血吸虫病防治知识、行为现状及健康教育需求情况,为制定科学有效的小学生血吸虫病健康教育方案提供参考依据。方法 采用分层整群抽样方法,抽取岳阳市城区洞庭湖湖畔学校和中心城区学校各2所,每所再从五、六年级分别抽取2个班的学生,通过调查问卷了解小学生血吸虫病相关知识、预防行为及健康教育需求,并进行统计分析。结果 共调查湖畔小学353人、中心城区小学363人,两组学校小学生年龄、性别、年级构成差异无统计学意义（t=-0.494,χ²_性别=1.615,χ²_年级=2.152;P均>0.05）。学生血吸虫病防治知识总知晓率为42.60%（305/716）,其中,湖畔学校小学生血防知识知晓率（52.97%）高于中心城区（32.51%）,差异有统计学意义（χ²=30.661,P<0.05）;学生行为正确率为76.68%（549/716）,血防知识知晓组行为正确率（81.31%）高于不知晓组（71.24%）,差异有统计学意义（χ²=6.384,P<0.05）。学生血防知识主要来源于老师（47.49%）;91.90%的学生愿意了解更多的血防知识,最喜欢的血防知识学习方式是参加课外活动（50.42%）。结论 岳阳市城区小学生的血防知识知晓率及行为正确率偏低,对血吸虫病防治健康教育需求较高。建议针对学生开展形式多样的血防健康教育,以增强学生的血吸虫病防护意识,提高自我防护能力。     

欧洲儿童肝豆状核变性诊疗推荐意见简介

正肝豆状核变性(Wilson’s disease,WD)是常染色隐性遗传相关的铜代谢障碍性疾病,估计患病率为1:30000。该病是由于编码P型ATP酶的ATP7B基因突变,影响铜的分泌及排泄所致铜沉积。儿童肝豆状核变性的临床表现可以从无症状肝脏疾病到肝硬化或者肝衰竭,但是神经精神症状却非常少见。常规的检测方法包括血清铜蓝蛋白、24 h尿铜。最终的诊断需要基于症状、评估铜代谢的生化学检     

A new concept of the anatomy of the anal sphincter mechanism and the physiology of defecation

The role of the external and internal anal sphincters in the mechanism of anal continence is presented. The external sphincter induces continence by 1) preventing internal sphincter relaxation, what I have called the “voluntary inhibition action,” and 2) mechanical compression of the rectal neck and anal canal proper. The mechanism of both actions is described. The internal sphincter plays a significant role not only in involuntary, but also in voluntary, continence. The importance of this role in the correction of anal incontinence is clarified. “Stress defecation,” a condition which follows internal sphincter damage, is discussed. A “single loop continence” theory is presented, based on the fact that each of the three loops of the external sphincter has its own innervation, attachment, and direction of muscle bundles; each loop thus acts as a separate sphincter. The clinical application of this theory is presented.