肾综合征出血热病毒单克隆抗体杂交瘤细胞系的建立与鉴定

用陕西省分离的肾综合征出血热(HFRS)病毒82-010H株免疫BALB/c纯系鼠和CxS/2小鼠,取免疫鼠脾细胞与SP2/O小鼠骨髓瘤细胞融合,获得10株分泌HFRS病毒特异性抗体的杂交瘤细胞系,对其中3H_4和4B_9两株单克隆抗体进行了鉴定。用各地分离的16株HFRS病毒抗原片与该两株单克隆抗体作了间接免疫荧光试验,结果表明:4B_9单克隆抗体只能与重型(黑线姬鼠型)HFRS疫区的病毒呈现阳性免疫荧光反应,而对轻型(褐家鼠及大林姬鼠型)HFRS疫区分离的病毒株不呈现免疫荧光反应;而3H_4单克隆抗体又能将重型和轻型HFRS毒株再行区分,这可能对HFRS病毒血清学分型和抗原分析以及流行病学调查研究等均有意义。     

肝病死亡病例多种组织的HBV-DNA和HBsAg检测(5例报道)

对血清学HBV感染标记阳性的5名肝病死亡病例,在进行尸检病理检查的同时,采集了肝脏以及肝外多种组织进行了HBV-DNA和HBsAg的检测。HBV-DNA检测采用斑点杂交技术(Dot blot hybridization),~(32)P HBV-DNA探针(Probe)由北京医科大学附属人民医院肝病研究室提供。组织HBsag的检测采用间接免疫荧光法。 5例肝脏组织的HBV-DNA检测均为阳性,3例并肝癌的癌组织亦为阳性。肝、脾、肾、腹腔淋巴结等组织的HBV-DNA阳性率均极高,胰、肺、皮肤次之。1例作了脊髓和视神经的HBV-DNA检测,亦为阳性,5例心脏组织均为阴性。5例中的4例作了血清HBV-DNA检测,仅1例阳性。5例中的3例以免疫荧光法检测HBsAg,肝、肾及肝癌组织均为阳性,而心、肺组织均为阴性。显微镜病理检查,肝、胰、肺组织均有明确的病理改变。 作者们提出HBV感染虽以肝脏为主,但却是一种多脏器感染。肝炎的肝外表现恐非单纯的免疫复合物所致。部分被诊断为内科肾脏疾患的病例,有可能实为肾脏的HBV感染,应予重视。     

免疫过氧化物酶技术在检测肾综合征出血热特异抗体中的应用

肾综合征出血热(HFRS)在我国流行广泛,目前血清学诊断主要依靠免疫荧光技术(IFA)。IFA需要昂贵的荧光显微镜和训练有素的工作人员,不便于在基层单位使用。我们将免疫过氧化物酶技术(IPA)间接法用于HFRS特异抗体的检测,并与IFA作了比较。结果发现IPA用于早期血清的检测,其检出的特异抗体滴度比IFA高2.6倍,用于恢复期血清检测,比IFA高20倍左右。在所检测的49份患者血清中,IPA检出阳性40份,IFA检出阳性35份,IFA阳性IPA阴性的仅1份。IPA敏感,特异性可靠,仅需普通光学显微镜,观察清晰,操作简单,便于基层单位使用。     

伯氏考克斯体新桥株感染BALB/c小鼠的实验研究

目的:分析伯氏考克斯体新桥株对BALB/c小鼠的感染性.方法:用伯氏考克斯体新桥株腹腔接种感染小鼠,于感染后7,14,21,28 d分别处死小鼠,采用伯氏考克斯体特异的荧光定量PCR检测感染小鼠肝、脾、肺组织样本中伯氏考克斯体含量.采用间接免疫荧光法检测感染小鼠血清特异性抗体水平.结果:感染后28 d内,小鼠肝、脾、肺组织中均检出大量伯氏考克斯体,第7天检出量最高,脾的含量显著高于肝和肺,肝的含量显著高于肺.随着感染时间延长,伯氏考克斯体检出量呈下降趋势,但血清特异性抗体水平则不断升高.结论:伯氏考克斯体新桥株为强毒株,可以引起BALB/c小鼠较长时间的稳定感染;BALB/c小鼠可以用作伯氏考克斯体感染模型.     

用VeroE-6细胞从病人血清中直接分离肾综合征出血热病毒

用引进的VeroE-6细胞株,接种肾综合征出血热(HFRS)病人的早期血清和全血标本,从中分离出21株与HFRS相关的病原体,分离阳性率达91.3％。用感染细胞制成的滴片标本     

四川南充肾综合征出血热高发区小兽动物带病毒状态的研究

1987年6月至1988年5月,我们对四川南充市郊肾综合征出血热(HFRS)高发区定时定点进行了小兽动物带病毒状态的研究。 方法:现场设点分为室内、室外和野外。室内指住室、厨房、畜圈等;室外指阶檐下5m以外,野外指院落外的菜地、稻田、平坝和坡地。采用夜间布夹法:室内按15m~2布夹1只,室外10m布夹1只,野外间隔10m布夹。应用间接免疫荧光法检测鼠肺HFRSV抗原,在无菌条件下剖取鼠肺,置于液氮罐保存,再作冰冻超薄切片。免疫荧光抗体购自上海生物制品研究所,抗76-118株兔血清为四川省防疫站惠赠,效价1∶1024;用76-118株阳性鼠脑抗原片作阳性对照,并设阴性对照。 结果;(1)小兽密度与种类组成:共布放     

酶联免疫法与金标法检测抗-HIV的比较

目的 观察ELISA(间接法、双抗原夹心法)快速诊断金标法(胶体金免疫分析技术)在检测抗-HIV中试剂质量比较,并分析其原因。方法 用2001年卫生部免疫检验室间质评HIV质控血清10份,其中6份为阳性血清(经Weet-Blot确诊),用两厂家生产的ELISA间接法、双抗原夹心法、金标法做比较。结果 两厂家生产的ELISA间接法一份阳性标本不能检出,但两厂家生产的ELISA双抗原夹心法和金标法阳性检出率为100％。结论 试剂的质量对保证大量的献血员、高危人群的HIV筛查工作至关重要。     

酶标抗体的制备及细胞内病毒和立克次体抗原的检出

用三种不同方法提取的抗体IgG与辣根过氧化物酶按戊二醛一步法和二步法制备酶标抗体。酶标抗体经免疫化学和免疫组织化学鉴定,初步认为可供光学显微镜检出细胞内病毒和立克次体抗原应用。乙型脑炎病毒感染小白鼠脑石蜡切片间接法酶标抗体染色的结果表明:脑干神经核和皮层海马回等部位的感染神经元的胞体、轴突及树突内均可检出棕黄色颗粒样抗原-抗体反应沉淀物。斑点热立克次体自然感染蜱血淋巴涂片的直接法和间接法酶标抗体染色均显示阳性反应。     

酶联免疫法与金标法检测抗-HIV的比较

目的 观察EUSA(间接法、双抗原夹心法)快速诊断金标法(胶体金免疫分析技术)在检测抗-HIV中试剂质量比较，并分析其原因。方法用2001年卫生部免疫检验室间质评HIV质控血清10份，其中6份为阳性血清(经Weet-Blot确诊)，用两厂家生产的EUSA间接法、双抗原夹心法、金标法做比较。结果两厂家生产的ELISA间接法一份阳性标本不能检出。但两厂家生产的EUSA双抗原夹心法和金标法阳性检出率为100％。结论试剂的质量对保证大量的献血员、高危人群的HIV筛查工作至关重要。     

建立流行性出血热(EHF)实验动物模型的研究 Ⅱ.应用免疫荧光法检查EHF病毒实验感染小鼠

流行性出血热(EHF)病毒通过传代可在小鼠乳鼠引起明显的症状。本文以免疫荧光作为判断指标,报告了感染小鼠体内EHF病毒抗原分布、血清抗体产生水平、病毒在体内繁殖动态等结果,并对流行性出血热的发病原理进行了探讨。材料和方法一、病毒、动物、感染方法参见前文报告。二、检查标本的取材及处理方法感染动物发病后,眼球放血致死,血清4℃保存备作抗体测定。横切鼠脑取视丘、海马、小脑、延脑等部位标本。内脏取肾、肺、心、肝、脾、肠以及胸腺、颌下腺等组织。皆切成大小约5×5×2mm的组织块,在95％乙醇     

Real-time PCR assay for the detection of picoplankton DNA distribution in the tissues of drowned rabbits

The detection of plankton DNA is one of the important methods for the diagnosis of drowning from postmortem tissues. This study investigated the quantities of picoplankton (Cyanobacteria) DNA in the lung, liver, kidney tissues and blood of drowned and non-drowned rabbits, and the sensitivity of detection of picoplankton DNA by polymerase chain reaction (PCR) detect for the diagnosis of death from drowning. For this purpose, the DNA of the 16S ribosomal RNA gene of picoplankton was quantitatively assayed from the tissues of drowned and non-drowned rabbits immersed in water after death. Each of the liver, kidney and lung tissues and blood were obtained from drowned and non-drowned rabbits. Picoplankton DNA in the tissues was extracted using the DNeasy® Blood & Tissue kit to determine the yield of picoplankton DNA from each tissue. TaqMan real-time PCR was performed for quantitative analysis of picoplankton DNA. Target DNA was detected in the liver, kidney and lung samples obtained from the drowned rabbits, while no picoplankton DNA was detected in the non-drowned rabbit tissues (except in lung samples). The results verified that direct PCR for the detection of picoplankton DNA is useful for the diagnosis of drowning. Although we observed seasonal changes in the quantity of picoplankton in river water, we were able to detect DNA from various organs of drowned bodies during the season when picoplankton were not the most abundant.     

枪伤后猪脑功能PET显像及各重要脏器病理学的研究

 目的探讨枪伤前后猪脑功能PET显像及各重要脏器病理学的改变.方法采用81式自动步枪,口径7.62mm,560式子弹,选取成年健康家猪左后肢肌肉丰厚处,避开大血管和骨头,射击距离5 m,射击后立即止血包扎,1 h后行脑功能PET检查,6 h后进行脑、心、肝、脾、肾和肺等脏器的病理学检查.结果枪伤前后脑功能葡萄糖代谢有明显差异(P＜0.01).枪伤后除肺脏有病理学改变外,其他重要脏器无明显病理学改变.肺组织主要表现为肺泡萎陷,含气量减少伴炎细胞浸润,肺水肿,肺泡结构不清,小血管淤血等.结论枪伤后猪脑PET功能显像代谢明显增加,肺部损伤明显,其作用机制与应激、局部周围组织炎性反应和远达效应有关.     

中国成年男子器官、组织中元素浓度研究

目的获得中国成年男子主要器官、组织中元素浓度代表值。方法作为系列研究第3阶段，在中国4个不同膳食类型地区，补充采集16例当地长住急死正常尸体肌肉、肋骨、肝、肾、肺、甲状腺、心、胃、脾、小肠、血液、皮肤、脂肪、睾丸、胰腺、大肠、肾上腺和胸腺共18种器官、组织样品，采用ICP-MS、ICP-AES和GF-AAS技术，测定56种元素浓度，与前两阶段相应结果合并统计取中位数作代表值。结果获得了这系列研究最多达68例中国成年男子尸体18种器官、组织样品中60种元素浓度代表值。结论除更新过去报道10种器官、组织50种元素浓度代表值外，首次获得新增8种器官、组织56种元素和已测10种器官、组织10种元素浓度代表值数据。这些结果为确定中国参考人器官、组织浓度参数参考值和在辐射防护领域其他应用提供了较前更全面和有代表性的国情依据。     

DIA3 phenotyping in human tissues,dental pulps,and hair roots by isoelectric focusing

Summary The polymorphism of DIA3 was investigated in tissues of various human organs, dental pulps, and hair roots by isoelectric focusing. DIA3 types were demonstrated from tissues of brain, prostate, testis, ovary, and uterus, but not from tissues of spleen, pancreas, heart, liver, muscle, lung, skin, and kidney. Determination was possible from dental pulps stored at room temperature for up to 2 weeks and from fresh hair roots. The results show that the DIA3 typing by isoelectric focusing is useful for medicolegal individualization of brain, reproductive organs, teeth, and hairs.     

52例中国人主要器官、组织中37种元素的浓度

目的测定中国成年男子主要器官、组织中元素浓度。方法在我国4个不同膳食类型地区补充采集21例急死正常尸体肌肉、肋骨、肝、。肾、肺、甲状腺、心、胃、脾和小肠样品，连同过去31例尸体采集的样品，采用ENAA、INAA、ICP-AES和ICP-MS方法及必要质控措施，共测定37种元素浓度。结果获得了我国成年男子这些器官、组织样品中37种元素浓度。结论除改善I在甲状腺和33种元素在其他5种器官、组织过去报道结果代表性外，本研究首次获得我国这些元素在心、胃、脾与小肠和Be、Cl、S、I在除甲状腺中Ⅰ以外10种器官、组织浓度资料。这些结果为确定中国成年男子器官、组织元素浓度背景值和中国参考人及生物动力学模型相应参数提供较前更为系统和代表性依据。本结果还和近年文献资料及ICRP参考人新估计值进行了比较。     

流行性出血热流产婴儿与胎盘的病理变化及巨噬细胞内抗原定位

妊娠6~ 个月孕妇感染流行性出血热(EHF),临床有典型的EHF的发病过程。第23病日血清学检测孕妇外周血EHF-IgG(卅)、IgM(-),第24病日流产一女婴,抽取脐带血检测EHF-IgG(卅),流产女婴心血检测EHF-IgG(卅),IgM(-)。病理:肾小球上皮细胞肿胀,上皮细胞足突融合、坏死,毛细血管基底膜皱缩,肾小管上皮细胞线粒体肥大,间质内大单核细胞浸润。肺间质细胞增生、水肿、毛细血管基底膜不规则增厚,肺泡壁Ⅱ型细胞增多。肝细胞肿胀,胞浆内查见包涵体样物质,有多核肝巨细胞,地依红染色阴性。胎盘母体面、脐带、肺、肝间质内大单核细胞、中性粒细胞浸润。EHF-MCAb荧光直接法染色、PAP、ABC等免疫组化染色,在巨噬细胞浆内均为阳性反应,从而证实母体感染EHF可以经胎盘感染给胎儿,其传播方式可能是通过巨噬细胞系统。     

52例中国人主要器官、组织中37种元素的负荷量研究

目的估算37种元素在中国成年男子主要器官、组织和全身的负荷量。方法在我国4个不同膳食类型地区补充采集21例急死正常尸体肌肉、肋骨、肝、。肾、肺、甲状腺、心、胃、脾和小肠样品，连同过去31例尸体所采集样品对37种元素的测定浓度，按中国参考人器官、组织重量参考值估算了相应器官、组织和全身的负荷量。结果获得了我国成年男子10种器官、组织中37种元素器官、组织和全身的负荷量。结论首次获得的这些结果为确定中国人器官、组织所测元素负荷量背景值和中国参考人相应负荷量参考值及生物动力学模型参数提供较前更为系统和有代表性的依据。     

Unpredictable tensile strength biomechanics may limit thawed cadaver use for simulant research

A series of tensile strength experiments were conducted using an un-embalmed elderly 91-year-old female cadaver, who had been frozen and thawed on five occasions. One sample was taken from the heart, kidney, oesophagus, skeletal muscle, ascending aorta, trachea, spleen, liver, lung, pancreas, pericardium, skin (abdomen) and skin (thorax) and the tensile strengths were measured using universal test equipment – Hounsfield H50KM (Hounsfield Test Equipment Ltd, Surrey, UK). Tensile strengths were: heart 34.9 Pa, spleen 45.6 Pa, kidney 100.7 Pa, liver 106.2 Pa, pancreas 148.9 Pa, oesophagus 216.5 Pa, skeletal muscle 288.9 Pa, lung 293.6 Pa, ascending aorta 588.2 Pa, pericardium 1341.9 Pa, trachea 1523.9 Pa, skin (abdomen) 3483.9 Pa, and skin (thorax) 3999.9 Pa. Compared with published values for fresh cadavers (that are only available for certain tissues), some organs and tissues had tensile strengths that fell well below the normal range (heart, oesophagus and ascending aorta), compared with others where the tensile strengths were well above the normal range (kidney and skeletal muscle). Only one tissue, from the trachea, fell within the normal range. The remainder of the data gave relative tensile strengths of other organs and tissues with the spleen being the least, and skin being the most, elastic tissue. Freezing and thawing cadaveric organs and tissues may alter their physical properties in ways that are not predicable, with both increases and decreases in tensile strength. Although this pilot study allows the relative tensile strengths of such tissues to be compared, it also demonstrates that physical changes following freezing and thawing alter properties in such a way that its usefulness as a simulant for normal tissues may be limited.     

32P-磷酸铬间质给药在荷人胰腺癌裸鼠的体内生物学分布及形态学表现

目的 研究^32P-磷酸铬(^32P胶体)瘤体间质给药治疗BALB／c—nu／nu裸鼠荷人胰腺癌(Pc-3)移植瘤时在体内相应组织中的分布、药代动力学特点及全身毒性反应。方法 51只荷瘤裸鼠，经瘤体给予不同剂量^32P胶体或尾静脉给药，分批处死，动态观察^32P胶体在裸鼠体内放射性分布和组织器官形态学表现，观察体重变化和计数WBC和PLT，测量瘤体表面放射性计数率。结果 ^32P胶体瘤体间质注射后其放射性计数率明显高于其他器官组织，器官组织放射性计数率瘤体给药明显低于尾静脉给药。增体给药有效半减期为13d。形态学检查显示给药后大部分Pc-3细胞被破坏，并出现分化较好的瘤细胞；肝、脾、肺及淋巴结等重要器官组织的辐射损伤为可逆性．未见明显骨髓抑制现象。结论 ^32P胶体瘤体间质给药是治疗胰腺癌安全、简便、有效的核素介入疗法。     

Autofluorescence spectroscopy in whole organs with a mobile detector system

RATIONALE AND OBJECTIVES: Autofluorescence can be exploited to obtain spectroscopic information about tissues or organs in a noninvasive fashion. The knowledge of normal organ patterns is a prerequisite for subsequent characterization of pathological states, eg, inflammation or tumors. Therefore, the aim of this study was to investigate the autofluorescence properties of healthy organs in mice. MATERIALS AND METHODS: Organs from C57Bl/6 mice were removed in toto and stored in physiologic sodium chloride solution on ice (non-perfused specimens). Investigations were performed with a custom-made mobile fluorescence detector. Excitation-emission matrices (EEMs) were measured in selected organs (bladder, brain, kidney, liver, and spleen) (n = 5). Afterwards, single-emission spectra were obtained in selected organs (bladder, colon, brain, kidney, liver, and spleen) and peak fluorescence signal intensities were calculated (n = 9). RESULTS: EEMs showed that excitation at wavelengths from 300-310 nm (emission spectra in all samples of bladder and brain; probably caused by collagen/elastin) and from 350-360 nm (emission spectra in all samples with the exception of spleen; probably caused by NAD(P)H) seem to be best suited for autofluorescence measurements in organs. The single-emission spectra measurements were noticeably different in terms of occurrence (yes/no response) and intensity of fluorescence emission peaks in different organs. CONCLUSION: Combined autofluorescence measurements of collagen/elastin (for structural information) and NAD(P)H (for functional information) allow conclusions about the target organs. Therefore, autofluorescence measurements seem to be a diagnostic tool feasible for characterization of tissue.