对虾病毒病和细菌合并感染的病理特点和诊断价值

应用电镜超薄切片及光学石蜡切和环氧树脂薄片技术,观察健康对照组,疾病始发组和濒临死亡组的草虾,中国对虾、日本对虾和肝胰腺和中肠。结果在３组４种对虾中均检测到病毒,只在濒临死亡组的４种对虾中发现病毒与细菌的合并感染。合并染的病毒有ＭＢＶ,球形病毒和淋细胞核型杆状病毒。细菌以弧菌为主。其病理变化表现为器官和组织的保护层受损、溶解样坏死和凝固样坏死,仅以病毒和细菌合并感染徐为养殖对虾病毒病早期诊断的指标     

养殖对虾病毒致病性的初步研究

用从日本对虾分离的杆伏病毒扣长毛对虾分离的球伏病毒分别感染健康无病毒的日本对虾,对虾死亡率分别为６５％（杆状病毒组）和５０％（球状病毒组）,而对照组的死亡率仅为２５％,在诸环境因素中,温度的上升和低压可能是对虾病毒病发作的最重要诱因之一。     

对虾白斑综合症病毒囊膜蛋白VP19的融合表达及其抗病毒感染作用

根据GenBank上WSSV囊膜蛋白基因vp 19的序列,设计并合成引物,PCR扩增得到vp19基因并克隆到pGEM‐T载体中,经过BamHⅠ/Hind Ⅲ酶切、连接并将vp19插入到pET32b表达载体中.用重组质粒pET32b-vp19转化大肠杆菌Origami(DE3)pLysS,在IPTG诱导下,融合蛋白Trx-VP19以可溶性的形式得到表达,经SDS-PAGE和Western-blot检测显示其分子量与预期的大小相符合.目的蛋白经Ni2+柱纯化并定量后分别直接注射鳌虾和包被饲料投喂鳌虾.实验结果表明注射Trx-VP19可以提高鳌虾个体抗WSSV感染力的作用.     

对虾白斑病毒1197基因的表达和产物纯化的研究

1993年以来,我国沿海省份相继发生养殖对虾大批死亡现象,损失严重,引起国内外研究者的重视.经过一段时间的努力研究后,我们报导了引起该病的病原是一种无包含体的杆形病毒[1].此后,该病毒的一些其他特性及引起对虾的病理学及组织学的变化又陆续见诸于报导[2,3].前文中我们报导了该病毒的一个早晚期基因1197的克隆和序列分析,并针对此基因设计了二个核酶,在体外成功地对此基因进行了切割[4].本文报导我们对该基因进行了原核表达,并对表达产物进行了纯化的研究,以期为阐明该基因表达产物的功能打下进一步研究的基础.     

细小病毒B19感染实验室诊断技术的研究进展

人类细小病毒感染,临床可有多种表现,尤其可导致胎儿早产、流产、死胎、贫血、发育迟缓及形成积水型胎儿。本文拟从电镜检查、ＥＬＩＳＡ、固相放射免疫技术（ＲＩＡ）、分子杂交及多聚酶链反应（ＰＣＲ）等几个方面对该病毒感染实验室诊断的国外最新研究进展作一介绍。     

对虾白斑综合症病毒囊膜蛋白VP19的融合表达及其抗病毒感染作用

根据GenBank上WSSV囊膜蛋白基因vp19的序列,设计并合成引物,PCR扩增得到vp19基因并克隆到pGEM‐T载体中,经过BamHⅠ/HindⅢ酶切、连接并将vp19插入到pET32b表达载体中。用重组质粒pET32b-vp19转化大肠杆菌Origam(iDE3)pLysS,在IPTG诱导下,融合蛋白Trx-VP19以可溶性的形式得到表达,经SDS-PAGE和Western-blot检测显示其分子量与预期的大小相符合。目的蛋白经Ni2 柱纯化并定量后分别直接注射鳌虾和包被饲料投喂鳌虾。实验结果表明注射Trx-VP19可以提高鳌虾个体抗WSSV感染力的作用。     

双抗体夹心ELISA法检测养殖对虾病毒的研究

用纯化的长毛对虾球状病毒（PPSV）和日本对虾中肠腺坏死杆状病毒（BMNV）制备新西兰兔抗BMNV和抗PPSV抗血清及Balb／c小鼠抗BMNV和抗PPSV抗血清,建立检测PSV和BMNV的双抗体夹心ELISA检测法,结果表明,双抗体夹心ELISA法具有较高的灵敏度,可以从100μl待测组织匀浆液中检测到50ng的PPSV蛋白,以及100ng的BMNV蛋白。不同病毒抗血清无交叉反应性,用该ELISA技术检测养殖对虾和多种采自养殖虾池及其附近的近海岸生物,发现相当比例的外观正常的对虾和近海岸生物已呈阳性反应     

肠杆菌科细菌致恶性肿瘤患者肺部感染的临床特点与耐药性分析

目的探讨肠杆菌科细菌致恶性肿瘤患者肺部感染的临床特点、病原学构成和耐药特征,为今后的临床治疗提供参考。方法收集2009年7月至2014年6月本院住院恶性肿瘤患者下呼吸道标本,应用BD Phoenix100全自动微生物鉴定仪鉴定菌种,药敏试验采用纸片法(K-B)进行,并进行ESBLs检测,按CLSI标准判定药敏结果,用WHONET 5.6软件分析数据。结果从恶性肿瘤患者下呼吸道标本中共分离肠杆科细菌256株,主要为肺炎克雷伯菌和大肠埃希菌,分别占39.5%和33.2%;药敏试验表明常见肠杆菌科细菌除对碳青霉烯类、含酶抑制剂抗菌素耐药率较低外,对其余抗菌药物均为中高度耐药;肺炎克雷伯菌的ESBLs检出率最高为60.4%。结论从本院恶性肿瘤合并下呼吸道感染患者中分离出的肠杆菌科细菌多重耐药现象严重,产酶率高,治疗该类细菌感染,首选碳青霉烯类、含酶抑制剂抗菌素,临床应加强ESBLs检测和耐药性监测,防止多重耐药菌的产生。     

甲烷和氢呼气检测诊断小肠细菌过度生长的现状

小肠细菌过度生长的传统诊断方法是小肠抽吸液培养。然而,此方法受到抽吸和培养技术的影响,并且受到标本对污染的敏感性的限制,迫切需要寻找一种切实可行的检测方法来填补临床空白。底物被人体口服后进入消化道,根据服用后时间,可以判别底物是否到达小肠或大肠。甲烷和氢气只来源于肠道细菌发酵的底物,这些气体最终通过肠道吸收入血并经肺排出,成为特定疾病的生物标志物。甲烷和氢呼气检测通过检测服用底物一段时间后口呼气中甲烷和氢气的浓度来判定小肠细菌是否过度生长。本文就小肠细菌过度生长诊断中检测气体的种类、检测前的准备工作、底物剂量、诊断时间和诊断阳性阈值等作一综述。

     

宫腔镜与阴道超声用于诊断子宫内膜病变的临床价值探讨

目的:探究阴道超声与宫腔镜对诊断子宫内膜病变的临床应用价值。方法:选取我院妇产科收治的子宫内膜病变患者153例,进行阴道超声及宫腔镜检测,观测患者病变类型以及病变部位,比较术后患者临床资料与术前阴道超声以及宫腔镜的检测结果,对阴道超声及宫腔镜的准确率、敏感性、阳性预测率以及阴性预测率等进行比较。结果:阴道超声检测内膜增生漏诊3例、内膜息肉误诊3例、粘膜下肌瘤误诊2例、内膜炎漏诊2例、内膜癌漏诊1例;宫腔镜检测内膜息肉误诊1例、黏膜下肌层误诊2例、内膜炎漏诊1例;与病理活检结果的符合率比较,宫腔镜的总符合率显著高于阴道超声,差异有统计学意义(P0.05)。宫腔镜检测子宫内膜病变的准确率、敏感性、阳性预测率及阴性预测率显著优于阴道超声,差异有统计学意义(P0.05)。结论:阴道超声和宫腔镜均可检测子宫内膜病变,前者操作简单、创伤小,后者操作复杂,但检出率、准确率及敏感度均较高,可减少临床误诊、漏诊的发生率。     

Virus diseases of farmed shrimp in the Western Hemisphere (the Americas): a review

Penaeid shrimp aquaculture is an important industry in the Americas, and the industry is based almost entirely on the culture of the Pacific White Shrimp, Litopenaeus vannamei. Western Hemisphere shrimp farmers in 14 countries in 2004 produced more than 200,000 metric tons of shrimp, generated more than $2 billion in revenue, and employed more than 500,000 people. Disease has had a major impact on shrimp aquaculture in the Americas since it became a significant commercial entity in the 1970s. Diseases due to viruses, rickettsial-like bacteria, true bacteria, protozoa, and fungi have emerged as major diseases of farmed shrimp in the region. Many of the bacterial, fungal and protozoan caused diseases are managed using improved culture practices, routine sanitation, and the use of chemotherapeutics. However, the virus diseases have been far more problematic to manage and they have been responsible for the most costly epizootics. Examples include the Taura syndrome pandemic that began in 1991-1992 when the disease emerged in Ecuador, and the subsequent White Spot Disease pandemic that followed its introduction to Central America from Asia in 1999. Because of their socioeconomic significance to shrimp farming, seven of the nine crustacean diseases listed by the World Animal Organization (OIE) are virus diseases of shrimp. Of the seven virus diseases of penaeid shrimp, five are native to the Americas or have become enzootic following their introduction. The shrimp virus diseases in the Americas are increasingly being managed by exclusion using a combination of biosecurity and the practice of culturing domesticated specific pathogen-free (SPF) stocks or specific pathogen-resistant (SPR) stocks. Despite the significant challenges posed by disease, the shrimp farming industry of the Americas has responded to the challenges posed by disease and it has developed methods to manage its diseases and mature into a sustainable industry.     

Detection of honey bee (Apis mellifera) viruses with an oligonucleotide microarray

In recent years, declines in honey bee (Apis mellifera L.) colonies have been observed to varying degrees worldwide with the worst losses in the USA being termed Colony Collapse Disorder (CCD). Pathogen load and the prevalence of honey bee viruses have been implicated in these losses and many diseased hives have multiple viruses present. We have designed and tested an oligonucleotide microarray which enables the simultaneous detection of nine honey bee viruses: Acute bee paralysis virus, Black queen cell virus, Chronic bee paralysis virus, Deformed wing virus, Kashmir bee virus, Sacbrood virus, Israel acute paralysis virus, Varroa destructor virus 1 and Slow paralysis virus. The microarray can be used to robustly diagnose nine viruses in one test.     

Infectious hypodermal and hematopoietic necrosis,a newly recognized virus disease of penaeid shrimp

Populations of the Pacific blue shrimp, Penaeus stylirostris, reared at the University of Arizona's experimental shrimp culture facility on Oahu in Hawaii from late 1980 through 1981, were severely affected by a highly acute and lethal disease of viral etiology. Also found to be susceptible to the disease were P. vannamei and P. monodon. The disease was named infectious hypodermal and hematopoietic necrosis (IHHN) disease to describe the principal lesions observed. The histopathology of acute and subacute IHHN disease in these species was dominated by the presence of conspicuous eosinophilic intranuclear-inclusion bodies of the Cowdry type A variety in ectodermally (especially the cuticular hypodermis) and mesodermally (especially the hematopoietic tissues) derived tissues that were undergoing necrosis. Electron microscopy of affected tissues demonstrated the presence of two or three types of virus-like particles with cubic morphology and diameters of 17 to 27 nm that suggest IHHN virus to be either a parvo- or picornavirus.     

Characterization of brown spot disease of gulf coast shrimp

Adult penaeid shrimp were collected from mariculture raceways, holding tanks, and near-shore waters of Galveston Island, Texas. Those with darkly pigmented lesions were chosen for study. When observed by scanning electron microscopy, diseased cuticle shows dense populations of bacteria within a lesion, whereas normal cuticle shows a relatively low number of bacteria. Some lesions exhibited a morphologically homogeneous population of bacteria while others revealed a heterogeneous or mixed infection. Bacteriological analysis of a homogenate of each lesion revealed the presence of a wide variety of organisms which produced extracellular lipases, proteases, and chitinases. Lipolytic and proteolytic organisms were consistently isolated from all 20 lesions examined, while chitinoclastic bacteria were found in all but two lesions. According to the presumptive isolation scheme used, 10 distinct phenotypes of bacteria were differentiated among the isolates from 20 lesions. These phenotypes were classified into four genera: Vibrio, Alteromonas, Spirillum, and Flavobacterium. Groups of 10 shrimp were abraded and inoculated with representative isolates from each of the 10 phenotypes. Four isolates were able to initiate lesion formation. These were two Vibrio species, one Alteromonas species, and one Spirillum species. All of these four pathogens were lipolytic, while only two were chitinoclastic. This evidence suggests that there is not one specific chitinoclastic bacterium which causes brown spot symptoms, but that a variety of bacteria acting alone or in groups may be the causal agents.     

对虾病毒病研究现状

对虾病毒病已严重危害世界对虾养殖业的发展,引起国内外学者的广泛关注。对近几年来对虾病毒病的研究现状和动态进行了概述,重点介绍了杆状病毒科(Baculoviridae)、细小病毒科(Parvoviridae)、呼肠孤病毒科(Reoviridae)、虹彩病毒科(Iridoviridae)、小RNA病毒科(Picornaviridae)、弹状病毒科(Rhabdoviridae)、被膜病毒科(Togaviridae)等几个科15种对虾病毒的特征、临床症状、病理特征、宿主范围和主要检测方法;对病毒病的传播途径、防治措施和存在的问题也进行了简要介绍。     

超声造影在肝硬化合并肝内局灶性小病灶中的诊断价值

目的:探讨超声造影(CEUS)在肝硬化合并肝内局灶性小病灶(≤3.0 cm)中的诊断价值。方法:选择我院自2010年4月至2014年8月患有肝硬化合并肝内局灶性小病灶的患者100例,采用MRI、CT及病理确定病灶良恶性,同时对比影像结果,计算常规超声和CEUS的准确性。结果:MRI、CT及病理确定病灶105个,其中恶性组45例,良性组60例,超声造影技术能清晰的显示出血流灌注的情况以及动脉相、门脉相及延迟相的特点。数据统计结果显示常规超声和CEUS的准确性、特异性和敏感性分别为59.05%、77.78%、45.00%和95.24%、95.56%、95.00%,两者之间的差异具有统计学意义(P0.05)。结论:相对于常规超声,CEUS对患有肝硬化合并肝内局灶性小病灶的患者具有更高的诊断价值。     

新型戊型肝炎诊断试剂盒的研制及其应用

用ＨＥＶＯＲＦ３合成肽及ＯＲＦ２重组抗原研制成新型ＨＥＶＥＩＡ诊断试剂盒。与ＧｅｎｌａｂｓＨＥＶＥＩＡ检测比较,灵敏度和特异性均达１００％（６０／６０）。三批试剂精密性测定均＜１０％。该试剂盒置４℃８个月或３７℃４ｄ保持稳定。检测不同肝炎患者ＨＥＶ抗体,发现急性非甲非乙非丙肝炎中有６３．２％,甲肝有１３．４％,乙肝有８．３％,丙肝有６．６％,正常人群为２．９％。所研制的戊型肝炎诊断试剂盒,灵敏度高,特异性强,精密性好,稳定性合格。适用于戊型肝炎诊断及戊肝病毒感染的流行病学调查     

戊型肝炎病毒与HBV重叠感染患者的血清学分析

目的了解戊型肝炎病毒(HEV)感染情况以及乙型肝炎病毒(HBV)重叠感染戊型肝炎病毒的血清学特点及临床意义。方法对HEV、HBV感染以及HBV重叠HEV感染进行血清学检测并进行统计学分析。结果 HBV重叠感染HEV病例多分布青壮年,且女性多于男性;重叠感染患者的年龄高峰都在21~40岁;HEV感染组与HBV-HEV重叠感染组比较,ALP、TB IL存在明显差异,胆汁淤积增多,黄疸程度加深,肝细胞损伤明显。结论 HEV主要侵犯青壮年,HBV重叠HEV感染后肝细胞损害加重,病情趋向重症化。     

血清抑制素-A表达联合经腹部超声对宫外孕的诊断价值分析

摘要 目的：探讨血清抑制素-A表达联合经腹部超声对宫外孕的诊断价值。方法：选取我院2018年1月到2021年8月共收治的60例宫外孕患者作为研究对象,将其分为观察组,另取同期来我院体检的60例正常妊娠孕妇作为对照组,对所有孕妇均行血清抑制素-A（INH-A）、碱性磷酸酶（ALP）、β-人绒毛膜促性腺激素（β-HCG）检测,并对所有孕妇进行经腹部超声诊断。对比两组患者血清中ALP、β-HCG与INH-A表达水平,对比两组孕妇经腹部超声诊断结果,分析两组孕妇血清INH-A和经腹部超声单一诊断与联合诊断对宫外孕的诊断价值。结果：观察组孕妇ALP、β-HCG水平明显低于对照组,INH-A水平明显高于对照组（P＜0.05）;观察组孕妇无胎心搏动、宫内无孕囊、附件区有包块以及子宫内膜厚度小于10 mm情况明显高于对照组（P＜0.05）;联合诊断检出率明显高于血清抑INH-A和经腹部超声（P＜0.05）,联合诊断的误诊率和漏诊率明显低于血清INH-A和经腹部超声（P＜0.05）;联合诊断的准确度、敏感度明显高于其他两组单一检查方法（P＜0.05）,三种诊断方法在特异度、阳性预测值和阴性预测值方面的检测价值对比并无显著差异（P＞0.05）。结论：应用血清抑制素-A表达联合经腹部超声诊断可提升宫外孕诊断的准确度与敏感性,因此可考虑对临床症状疑似宫外孕的患者应用两者联合诊断,提升诊断准确率。