人生长激素的生物素—新和素放大酶联免疫法的建立

为建立有生物素和亲和素放大系统的人血清生长激素 (hGH)酶联免疫分析法 (BA ELISA) ,以微量的GH抗原免疫BALB/C小鼠 ,利用杂交瘤技术制备出 2 9株分泌抗hGH单克隆抗体 (MCA)的杂交瘤株。选择最佳配对MCA ,利用亲和素与生物素高亲和力的特性 ,建立了血清hGH双位点的BA ELISA法。我们的MCA滴度为 (0 5～ 5 0 0 )× 10 4,亲和常数Ka =(0 13～ 1 40 )× 10 10 L/mol。血清hGH的ELISA的灵敏度为 0 0 4± 0 0 1μg/L ;在血清中分别加入 1,2 5 ,5 μg/L的hGH ,其回收率为 90 7%～ 10 7 6 % (平均为 10 1 2 0 %± 0 0 3 % ) ;hGH含量为 2 8、10 0、2 9 8μg/L的血清批内、批间变异系数分别为 4 9%、3 8%、7 9%和 6 2 %、3 5 %、9 5 % ;用本法测定了正常儿童、生长激素缺乏症 (GHD)患儿和活动性肢端肥大症患者空腹血清hGH水平 ,分别为 1 87± 3 0、0 30± 0 42和 8 71± 6 95 μg/L。上述结果表明 ,本ELISA方法灵敏、特异、稳定、准确 ,能确切反映hGH分泌功能 ,在诸多方面均优于RIA。     

CA125时间分辨免疫荧光分析法及其临床应用

以 CA1 2 5单克隆抗体包被微孔板 ,用平衡饱和法与 CA1 2 5、Eu3+标记异硫氰酸苄基二乙烯三胺四乙酸 -CA1 2 5单抗形成三层夹心 ,以β-二酮体为增强液 ,建立 CA1 2 5的时间分辩免疫荧光分析法 ( IFMA)。方法的批内和批间 CV分别为 2 .4 5%、2 .66% ,平均回收率 1 0 4 .2 9% ,灵敏度 3.2 7U/L,可测范围为 6.0 5-2 0 62 U/L。本法与 CEA无交叉 ,与 AFP有 6.77%的交叉 ,与β-HCG有 1 0 .89%的交叉。 84名健康女性血清 CA1 2 5浓度为 1 1 .4 2± 1 2 .2 0 U/L,34例卵巢癌患者血清 CA1 2 5浓度为 334 .33± 376.66U/L,与对照组比较有高度显著性差异 ( P<0 .0 1 )。血清样品测定结果与酶免疫分析 ( ELISA)和化学发光免疫分析 ( CLIA)测定结果呈相关 ,其相关系数 ( r)分别为 0 .888和 0 .899。诊断试验结果表明 ,CA1 2 5IFMA的敏感度、特异性、准确度和预测值等指标符合临床诊断要求     

微粒子化学发光免疫测定促甲状腺素敏感性方法的建立与应用

目的 :建立一种低成本、高敏感性检测血清促甲状腺激素(TSH)的微粒子化学发光免疫分析 (CLEIA)方法。方法 :采用碱性磷酸酶标记的抗TSHβ亚单位特异性单克隆抗体 (mAb) ,与FITC偶联的抗TSHα亚单位的另一mAb配对 ,构成双抗体夹心免疫结合 ,用抗FITC多抗免疫磁珠做固相分离载体 ,用金刚烷胺CSPD为发光底物的非均相免疫分析法。结果 :方法灵敏度 4 μIU/L ,批内变异系数 (CV)平均为7.4 5 % ,批间CV平均为 10 .4 5 % ,回收率为 91.4 %～ 10 2 .4 %。将实际样品测定与ACS 180  系统检测的结果相比较有较好的相关性。结论 :微粒子化学发光定量测定TSH的成本低、灵敏度及特异性高和稳定性好 ,具有广阔的应用前景     

NGF放射免疫分析及其应用

目的 :建立神经生长因子 (NGF)放射免疫分析 ,并初步探讨其应用价值。方法 :将NGF免疫新西兰家兔制备抗体。建成NGF放射免疫分析法 ,研制成功放免试剂盒。应用本试剂盒测定 81例抑郁症患者血浆和 32例正常人血浆 ,4 0例小鼠颌下腺组织中NGF水平。结果 :灵敏度为 1 0 pg/ml。抗血清特异性 ,与GDNF、EGF、CNTF、β -EP、CCK、GHRH、GH、BNP、Bombesin等均无交叉反应。批内变异系数 <3 85 % (n =1 0 ) ,批间变异系数 <5 1 7% (n =5 )。抑郁症患者血浆NGF浓度为 36 0 0 5± 5 1 89pg/ml,正常为 31 3 5 3±34 0 2 pg/ml。小鼠颌下腺组织中NGF浓度为 2 0 2 0 9± 32 6 1ng/mg。 结论 :NGF放射免疫分析灵敏度高、操作简便 ,为进一步了解NGF在生理和病理状态下的作用提供了可靠的方法     

PSA、fPSA双标记时间分辨荧光免疫分析临床应用价值

目的 :探讨PSA、fPSA双标记时间分辨荧光免疫分析 (DELFIA)技术在前列腺良恶性疾病诊断中的价值。方法 :采用双标记DELFIA检测 2 3例病理确诊的前列腺癌 (PCa) ,4 0例良性前列腺增生 (BPH)及30例正常对照血清PSA、fPSA水平 ,并与化学发光分析 (Immulite)比较。结果 :PSADELFIA测定结果PCa组(1 5 8 39± 92 5 5ng/ml)高于BPH组 (9 0 0± 1 0 6 1ng/ml)及对照组 (1 1 7± 0 6 9ng/ml) ,三组间比较有极显著性差异 (p <0 0 0 1 )。DELFIA与Immulite两种方法测定结果呈高度正相关关系 (r =0 95 4 ,p <0 0 0 1 )。PSADELFIA敏感性为 1 0 0 % ,特异性为 5 5 0 % ;Immulite敏感性为 1 0 0 % ,特异性为 4 7 5 %。fPSADELFIA测定结果PCa组 (73 8± 93 5 8ng/ml)高于BPH组 (1 5 8± 1 6 7ng/ml)及对照组 (0 31± 0 1 7ng/ml) ;三组间比较均有极显著性差异 (p <0 0 0 1 )。BPH组及对照组两种方法测定结果呈正相关关系 (r =0 94 ,p <0 0 0 1 ) ,PCa组两种方法测定结果亦呈正相关关系 (r =0 5 6 3,p <0 0 1 )。 结论 :PSA、fPSA双标记时间分辨荧光免疫分析一次检测可同时测出PSA及fPSA结果 ,这种分析对前列腺癌的检测有很高的敏感性     

C型利钠肽放免试剂盒研制及初步临床应用

目的 :建立C型利钠肽 (CNP)放射免疫分析 ,并初步探讨其临床应用价值。方法 :将甲状腺球蛋白与CNP联接 ,免疫家兔制备抗体。建成CNP放射免疫分析法 ,研制成功放免试剂盒。应用本试剂盒测定83例健康人、5 4例冠心病、2 5例心力衰竭、73例高血压患者血浆CNP水平。结果 :分析灵敏度为 5pg/ml。所获CNP抗血清特异性强 ,与心钠素 (ANP)、神经降压素 (NT)、神经肽Y(NPY)、内皮素 (ET)、降钙素基因相关肽(CGRP)、降钙素 (CT)等多肽均无交叉反应。批内变异系数 <10 % ,批间变异系数 <15 %。心力衰竭、冠心病、高血压患者血浆CNP浓度分别为 6 2 5pg/ml、4 7 9pg/ml、5 6 4pg/ml,明显高于健康人 (2 7 5pg/ml)。结论 :CNP放射免疫分析操作简便、快速 ,为进一步研究CNP在人体生理、病理状态及基础医学研究工作提供了一种可靠的手段。     

758例住院躯体疾病患者焦虑抑郁自评结果分析

Dunbar通过对各类躯体疾病病人的心理社会问题研究 ,提出心理因素可以影响疾病的发生、病程及预后[1 ] 。情绪障碍是影响躯体疾病患者康复的重要因素。本文选择丰台区 3所区属综合性医院 (南苑医院、长辛店医院、铁营医院 )2 0 0 0 10～ 2 0 0 2 3内科住院病人共 75 8例进行了分析。 75 8例患者平均年龄 5 9±10岁 ( 2 4-87) ;男性 42 5例 ( 5 6 0 7% ) ,平均 5 9± 10岁 ,女性 3 3 3例 ( 4 3 93 % ) ,平均 5 9± 10岁 ;高血压 2 5 9例 ( 3 4 2 % ) ,冠心病 2 0 4例 ( 2 6 9% ) ,糖尿病 15 5例( 2 0 4% ) ,脑血管病 14 0例 ( 18 5 % ) …     

HCG放射免疫分析与电化学发光分析的比较

2 2　重复性测定　分别用两种方法对各自的质控 1、质控 2浓度样本进行 10个批次以上的HCG测定。RIA法批内变异系数分别为 8 6 2 %、8 73% ;批间变异系数分别为14 38%、9 0 9%。ECLIA法批内变异系数分别为 3 84 %、3 2 9% ;批间变异系数分别为 5 90 %、5 31%。3　讨论83份受检者用两种方法测出血清HCG结果的相关分析表明 ,放射免疫分析和电化学发光分析测定重复性均在允许范围内 ,重复性测定结果表明ECLIA法明显优于RIA法可达到批内CV <4 % ,批间CV <6 %。其原因可能为ECLIA法为全自动操作 ,保证各次实验条件一致 ,且方法学本身和试剂盒质量均优于RIA所致。RIA法系手工操作 ,易受操作者情绪、熟练程度等多方面的影响。两方法所测结果呈显著正相关 ,r =0 .84 9。ECLIA法未孕健康妇女HCG测值低于RIA法 ,而孕妇及患者则明显高于RIA法 ,ECLIA法最低可测值为 0 1U/L ,显示其灵敏度明显高于RIA法。两方法均能满足临床需要。ECLIA法灵敏、准确但价格昂贵。RIA法成熟、便宜、易于开展。     

β_2-m单克隆抗体放射免疫试验的建立

本文首次采用β_2-m单克隆抗体,建立了竞争性、宽范围的放射免疫测定法。本方法最小检出值0.27μg/管,批内变异系数为4.47～7.89%,批间变异系数为9.07～15%,回收率为95—105%。测定43例体检血清标本,同市售商品药盒多克隆抗体放免法相比较,两者之间有极显著正相关(p<0.001)。实验结果表明:β_2-m单克隆抗体完全可以替代多克隆抗体,且本法检测范围较宽(0.31～10μg/ml),缩小了样品稀释倍数,使检测方法更简便。     

染色体t(8;21)伴急性髓系白血病六例

病例1～4　男性3例、女性1例,年龄2 2～3 8岁。因发热、牙龈渗血就诊。查体:贫血貌,胸骨有压痛,肝、脾及淋巴结无肿大。外周血WBC 2 .2～3 .8×10 9/L ,BPC 5 .4～7.8×10 9/L ,Hb 4.5～9.0g/L ;分类:原粒 早幼粒0 .0 7～0 .10 ,幼单0 5 7～0 .62。骨髓象:原粒 早幼粒占2 0 %～2 6% ,原单 幼单占42 .2 %～48%。过氧化酶染色阳性或弱阳性,酯酶 氟化钠抑制试验,抑制前阳性率92 %～95 % ,积分176～182 ,抑制率3 1%～3 5 %。诊断为急性粒 单细胞性白血病(M 4b)。细胞遗传学检查:常规培养2 4～48h ,收获制片,G显带,分析计数2 0～3 0个中期…     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

The dominant diseases of modernized societies as omega-3 essential fatty acid deficiency syndrome: Substrate beriberi

About 1900, modern food selection and processing caused widespread $\text{epidemics}$ of the B vitamin deficiency diseases of beriberi and pellagra which, for genetic reasons, often expressed as different diseases ranging from bowel and heart disease to dermatoses and psychoses. But the B vitamins merely help convert essential fatty acids (EFA) into the prostaglandin (PG) tissue regulators and it now turns out that, through hydrogenation, milling and selection of w3-poor southern foods, we have also been systematically depleting, by as much as 90%, a newly discovered trace Nordic EFA (w3) of special importance to primates and sole precursor of the PG3(4) series, even as a concurrent fiber deficiency increases body demand for EFA. Since substrate EFA is processed by many B vitamin catalysts, an EFA deficiency will mimic a $\text{panh}$ypovitaminosis B, i.e., a mixture of $\text{substrate}$ beriberi and $\text{substrate}$ pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse $\text{endemic}$ disease. This would consitute a second stage of the Modern Malnutrition and explain why some workers now hold the dominant diseases of modermized societies to be new, nutritionally based, pellagraform yet lipid-related and to range, once again, from heart disease to psychosis. It is an assumption that our dominant diseases are unrelated to each other or are merely revealed by our diagnostic acumen and therapeutic success; and that hydrogenating millions of tons of food oils annually, to destroy the rancidity producing w3-EFA, is safe for $\text{primates}$. Extensive beriberiform disease is reported here in 32 typical cases taken from medical practice which responds strikingly to linseed oil supplements (60% w3-EFA) in confirmation of identical results in Capuchins.     

'Very sore nights and days': the child's experience of illness in early modern England, c.1580-1720

Sick children were ubiquitous in early modern England, and yet they have received very little attention from historians. Taking the elusive perspective of the child, this article explores the physical, emotional, and spiritual experience of illness in England between approximately 1580 and 1720. What was it like being ill and suffering pain? How did the young respond emotionally to the anticipation of death? It is argued that children’s experiences were characterised by profound ambivalence: illness could be terrifying and distressing, but also a source of emotional and spiritual fulfilment and joy. This interpretation challenges the common assumption amongst medical historians that the experiences of early modern patients were utterly miserable. It also sheds light on children’s emotional feelings for their parents, a subject often overlooked in the historiography of childhood. The primary sources used in this article include diaries, autobiographies, letters, the biographies of pious children, printed possession cases, doctors’ casebooks, and theological treatises concerning the afterlife.     

Guidelines for the Validation and Use of Immunoassays for Determination of Introduced Proteins in Biotechnology Enhanced Crops and Derived Food Ingredients

Recent advancements in agricultural biotechnology have created a need for analytical techniques to determine introduced proteins in crops enhanced through modern biotechnology techniques. These proteins are expressed in plant tissues and may be present in food ingredients. Immunoassays are ideally suited for protein detection and may be used as both quantitative and threshold methods. Microplate ELISA and lateral flow devices are two of the most commonly used immunoassay formats for agricultural biotechnology applications. This paper provides general background information and a discussion of criteria for the validation and application of immunochemical methods to the analysis of proteins introduced into plants and food ingredients using biotechnology methods. It is the result of a collaborative effort of members of the Analytical Environmental Immunochemical Consortium. This collaborative effort represents the combined expertise of several organizations to reach consensus on establishing guidelines for the validation and use of immunoassays. Further, the paper offers developers and users a consistent approach to adopting the technology as well as aid in producing accurate and meaningful results.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

Ultracryotomy: development and application (with examples from the yeast cytology) (author's transl)

The preparation steps usually necessary for obtaining ultrathin frozen sections of biological material (chemical prefixation, enclosing, cryoprotective treatment, freezing, sectioning, and post-staining the sections for transmission electron microscopy) are submitted to a critical analysis. The application of cryo-ultramicrotomy, in particularly for cytochemical purposes, is reviewed. Fundamental considerations of chemical prefixation and poststaining are supported by examples from yeast cytology. Furthermore, the efficiency of the cryo-ultramicrotomy (electron optical resolution of ultrastructural details) is demonstrated on yeast cells and protoplasts.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.