人类8型疱疹病毒开放读码框73C基因原核

目的构建人类8型疱疹病毒（HHV-8）基因开放读码框（ORF）73c重组原核表达质粒并诱导表达，获得HHV-8潜伏态相关核抗原重组蛋白，并对其免疫活性进行初步鉴定。方法软件设计引物，分别在引物两端添加特异的酶切位点，以pGEM-Teasy／ORF73质粒为模板，聚合酶链反应（PCR）扩增ORF73C基因序列，克隆人原核表达载体pET-28a（＋），构建原核表达质粒pET28a-ORF73c；转化E.coli DH5α，经酶切、测序鉴定其插入序列的正确性，转入E.coli BL21（DE3），并诱导表达，以多聚组氨酸亲合层析柱纯化，凝胶蛋白电泳、Western印迹方法行表达蛋白的分析和抗原特异性免疫鉴定。结果测序结果表明构建的pET28a-ORF73C原核表达质粒连接正确，插入的ORF73c基因片段为453bp。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS—PAGE）表明ORF73c基因成功表达，在相对分子质量20000处有表达条带；Western印迹分析显示重组蛋白能被卡波济肉瘤患者阳性血清识别，具有良好的抗原性。结论成功构建了pET28a-ORF73C原核表达质粒，获得的纯化HHV-8潜伏态相关核抗原蛋白与卡波济肉瘤患者血清具有特异性识别，显示其作为检测抗原的可能，但其敏感性和特异性则有待进一步的验证。     

戊型肝炎

戊型肝炎病毒ORF2抗原在抗．HEV ELISA检测中的应用——阮冰等(浙江杭州市浙江大学医学院附属第一医院卫生部传染病重点实验室310003)《感染病杂志》2004，2(4)：167-169[比较重组蛋白与合成肽抗原建立的酶联免疫吸附试验(ELISA)在戊型肝炎实验室诊断中的应用价值。方法：选择戊型肝炎病毒第二开放读码框架(ORF2)，分别采用重组蛋白及合成肽抗原，建立ELISA，同时检测97份戊型肝炎急性期患，47份甲、乙、丙、丁、庚型肝炎患及120份健康供血的血清抗．HEV。结果：重组蛋白ELISA、合成肽ELISA与Genelabs重组抗原ELISA试剂盒检测结果的总符合率分别为96．9％和92％；重组蛋白ELISA与合成肽ELISA的一致率为91％；     

人类8型疱疹病毒与胃肠型HIV卡波西肉瘤组织ORF42、mTORC1表达相关性研究

目的 探究胃肠型HIV卡波西肉瘤组织中人类8型疱疹病毒(HHV8)感染与ORF42、哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)表达的关系.方法 选取2015年9月至2019年9月新疆维吾尔自治区人民医院收治的艾滋病合并胃肠型卡波西肉瘤患者36例作为研究对象.留取入组患者卡波西肉瘤组织及瘤旁组织,采用免疫组织化学染色...     

HHV-8 ORF73C基因的克隆及其在大肠杆菌中的表达

利用PCR技术获得人疱疹病毒8型（HHV-8）ORF73C基因，克隆至pGEMT—easy载体，进-步克隆至原核表达载体pGEX-6P-1中，构建含HHV-8 ORF73C基因的重组表达质粒pGEXORF73；重组表达质粒pGEXORF73在大肠杆菌中得到有效表达。     

实时荧光定量聚合酶链反应检测人类8型疱疹病毒载量方法的建立及其诊断应用

目的 建立一种检测人类8型疱疹病毒(HHV-8)载量的SYBR GreenⅠ实时荧光定量PCR(RT-qPCR)方法,并进行初步诊断应用.方法 分别构建HHV-8开放阅读框(ORF)26和内参照基因β-actin重组T载体,制备质粒标准品.确定最佳实验条件,建立高灵敏度的RT-qPCR方法.并检测30份经病理学确诊的卡波西肉瘤(KS)石蜡组织中HHV-8载量,初步评价其应用的有效性.结果 明确了建立该法的最关键因素为退火温度和延伸环节.标准曲线参数良好,ORF26和β-actin标准品Ct值与其相应梯度稀释标准品浓度呈良好线性关系(γ2＞0.990),扩增效率分别为88.3%和90.3%.经熔解曲线和电泳分析,产物特异.30份KS样本中HH V-8 ORF26的检出率为100%.经典型KS中HHV-8载量显著高于AIDS相关型KS(均数分别为69.18和8.63,x2=7.950,P=0.005).结论 成功建立了灵敏度高的HHV-8实时荧光定量PCR检测方法,为临床诊断可疑或感染病例提供了技术平台.     

人类8型疱疹病毒在新疆地区献血员中的感染率分析

人类8型疱疹病毒(human herpesvirus 8,HHV-8)在北美、欧洲和大部分亚洲国家属低流行,性传播为主要传播途径[1].在中国,HHV-8所致卡波西肉瘤(Kaposi's sarcoma,KS)病例主要集中在新疆地区的维吾尔族和哈萨克族,确切传播方式不明.研究表明,新疆是HHV-8感染的流行区[2].Hladik等[3]提出HHV-8可经输血传     

梅毒螺旋体Tp0453特异性抗原优势表位区段的克隆表达及其在血清学诊断中的初步评价

目的 应用基因工程技术在大肠埃希菌中表达梅毒螺旋体(Tp) Tp0453(62～224位氨基酸)重组蛋白,为提高梅毒血清学诊断试剂的特异性提供实验基础.方法 采用PCR法从Tp全基因组中扩增目的片段Tp0453(第184～672位核苷酸),T-A克隆后构建原核表达质粒pQE30-Tp0453,酶切鉴定后转化E.coli M15,异丙基-β-D-硫代吡喃半乳糖苷诱导表达,Ni2+亲和层析柱纯化目的蛋白,表达产物经Western印迹鉴定其免疫反应性.以纯化的重组抗原建立双抗原夹心法ELISA,检测经Tp抗体明胶颗粒凝集试验(TPPA)法确证的梅毒阳性血清48份、阴性血清40份.结果 PCR法扩增出约490 bp目的片段,成功构建原核表达质粒pQE30-Tp0453,并在E.coli M15中表达,蛋白表达率为18％,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳初步测定目的蛋白的相对分子质量为21 000,破菌后电泳证实目的蛋白以包涵体形式表达,纯化后蛋白纯度＞95％,Western印迹证实该蛋白能与梅毒患者阳性血清发生特异性反应.双抗原夹心法ELISA检测88份临床标本,灵敏度为97.9％(47/48),特异度为100.0％(40/40),与TPPA的总符合率为98.9％(87/88).结论 所表达的Tp0453重组蛋白具有良好的免疫反应性,为开发临床检测效果更好的梅毒诊断试剂盒提供了实验基础.     

贝氏柯克斯体5种重组表面蛋白抗原包被酶联免疫吸附试验的特异性和敏感性分析北大核心CSCD

目的使用贝氏柯克斯体5种重组表面蛋白作为包被抗原的酶联免疫吸附试验用于血清学检测的特异性和敏感性分析。方法本研究建立由贝氏柯克斯体的Com1、GroEL、Mip、OmpH、RplL重组表面蛋白抗原分别包被的酶联免疫吸附试验(ELISA)方法,然后分别检测贝氏柯克斯体实验感染小鼠血清和Q热患者血清中IgG特异性抗体。结果 8份贝氏柯克斯体感染小鼠血清中除1份血清与RplL反应为阴性外其它均为阳性;11份Q热患者血清中除1份血清与RplL、1份血清与OmpH反应为阴性外,其它血清反应均为阳性。将这5种重组蛋白分别与莫氏立克次体、黑龙江立克次体、恙虫病东方体实验感染小鼠血清反应,除1份黑龙江立克次体感染小鼠血清与GroEL反应为阳性外,其它血清反应均为阴性。另外,Com1对斑疹伤寒、斑点热、恙虫病患者血清的阳性反应率平均为22.2%,GroEL为25.0%,Mip为25.0%,OmpH为25.0%,RplL为13.9%。结论这些结果证明这5种重组蛋白抗原均可被贝氏柯克斯体感染血清所识别。所建立的ELISA方法具有良好特异性和敏感性,可用作Q热血清学诊断。     

丙型肝炎

丙型肝炎病毒不同基因型NS3蛋白的抗原异质性分析——乔伟振等(江苏无锡市第一人民医院中心实验室214001)《中华微生物学和免疫学杂志》2005，25(1)：10-14（探讨不同基因型丙型肝炎病毒(HCV)NS3蛋白的抗原特性及其用于抗．HCV检测的意义。方法：分别构建和表达含有HCVl型和6型NS3基因片段的重组质粒和重组蛋白，以ELISA法和Westernblot分析不同基因型重组蛋白与已知抗．HCV阳性血清的抗原反应性。结果：HCV1型和6型NS3重组蛋白氨基酸序列的同源性为83．2％；85份抗-HCV阳性血清以此不同基因型HCVNS3单片段抗原检测，     

重组抗原检测戊型肝炎病毒IgM抗体及其意义

用戊型肝炎病毒(HEV)重组抗原建立酶联免疫吸附试验(ELISA)，检测肝炎患者和健康供血人群血清中抗戊型肝炎病毒IgM类抗体(抗-HEV IgM)，并评价其检测意义。在与合成肽抗原比较检测的77份急性戊型肝炎患者血清中，有54份(70.1%)由重组抗原捡测抗-HEV IgM阳性。其阳性率明显高于台成酞抗原(21/77,27．3％)。15例戊型肝炎患者双份血清用重组抗原ELISA检铡，急性期血清有11份抗-HEV IgM阳性，恢复期血清仅1份阳性。抗-HEV IgG阳性的健康供血者(8人)和甲、乙、丙型肝炎患者(11例)无1例IgM抗体阳性。结果表明，抗-HEV IgM可以作为戊型肝炎病毒新近感染的标志。HEV重组抗原检铡抗-HEV IgM敏感性高，特异性强，有助于戊型肝炎病毒感染的早期诊断。     

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.     

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.     

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.     

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.     

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.     

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。     

安徽巢湖市丝虫病控制、消除历程和监测报告

【摘要】 目的  回顾并总结巢湖市丝虫病的流行、控制,以及消除丝虫病的历程。 方法  统计分析1970~1989年在该市采取以消灭传染源、切断传播途径为主的防治策略,以及展开大规模防治的各个阶段。 结果  1989年,采用分层整群抽样调查方法调查,微丝蚴率下降至0.032%,以行政村为单位,微丝蚴率已控制在1.0%以下,达到卫生部规定的基本消灭丝虫病标准;1991年,经省级调查考核,认定全市已达到卫生部规定的基本消灭丝虫病标准。1996年,省级审评确认已达到卫生部颁布的消灭丝虫病标准。经过10年的监测和跟踪治疗,到2005年,全市丝虫病病原学监测未查见微丝蚴阳性者,解剖镜检人房内淡色库蚊1 348只,蚊体内均未查见人体幼丝虫。 结论  巢湖市达到了消除丝虫病的标准。     

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.