用基因工程表达的HBcAg制备抗-HBcAg单克隆抗体及其初步应用

用基因工程表达的NBcAg免疫BALB/c小鼠,取脾细胞与小鼠骨髓瘤细胞Sp2/0融合,以ELISA抑制法检测抗体,获得28株分泌抗-HBc McAb的杂交瘤细胞系。对其中7株进行了克隆化,培养上清抗体滴度1:320～1:2560,免疫小鼠腹水和血清滴度1:10万～1:80万。Ig类型测定2株IgG1、2株IgG2a、1株IgG3、2株IgM。杂交瘤细胞系在体外连续传代培养6个月及液氮冻存的细胞系经复苏后仍能稳定地分泌McAb。得到的McAb只与HBcAg反应且能被HBcAg特异阻断,证明其特异性高。用ELISA间接法测定了McAb的相对亲和力,从50％最大结合的浓度分析结果,7株McAb的相对亲和力为2E6>2F5>2B11>2C10>1A7>1B7>1H7,浓度范围为0.6～10μg/ml。2E6和2C10 2株McAb与HRP结合,用ELISA双抗休夹心法筛选出2F5、2B111和H7 3株McAb适合作为包被抗体。2F5与2E6-HRP配对,建立了快速McAb-ELIS抑制法测定患者血清中抗-HBc。用本法测定了222份临床患者血清标本,结果与“华美”试剂盒测定结果基本一致。     

抗A型和抗B型血型单克隆抗体的制备及其在血型鉴定中的初步应用

本试验用A型和B型人红细胞混合悬液免疫BALB/c小鼠,取免疫脾细胞与同系鼠Sp2/0骨髓瘤细胞融合,建立了5株分泌盐水凝集抗体的杂交瘤细胞株。3次克隆化后,细胞株稳定分泌McAb已6个月。经A、B、O各型红细胞鉴定及阻断试验、吸收释放试验结果证明:H_2、H_5、B_7株是特异性抗A型抗原的,D_5株是特异性抗B型抗原的。本试验制备的McAb亲合力高于人抗血清,用保护剂稀释后稳定性良好。用McAb和人抗血清对献血员血液标本692份,脐带血58份做血型鉴定所得结果完全一致。     

单克隆抗体亲和层析法纯化HFRS病毒血凝素抗原的研究

用蔗糖-丙酮法从HFRS病毒陈株感染的乳小白鼠鼠脑中提取粗制血凝素,将此粗制血凝素通过抗HFRS病毒血凝素McAb 3G_1与Sepharosc 4B偶联的免疫吸附柱,获得纯化HFRS病毒血凝素抗原具有较高的血凝滴度和良好的免疫原性。用SDS-PAGE和Western-blot技术分析该纯化的血凝素抗原,并与同法纯化的正常鼠脑抗原比较,证明该特异性血凝素抗原分子量为5×10~4道尔顿。用2株抗HFRS病毒以及10株抗HFRS病毒血凝素McAb,以ELISA阻断试验分析纯化50K血凝素,结果有10株McAb可阻断McAb3G_1与其结合,其抗原决定簇之间有部分相同或童叠,提示这些具有不同特性的抗原决定簇确实位于同一蛋白上。以上结果表明,该50K血凝素蛋白特性及结构均较复杂,尚待进一步研究。     

巨细胞病毒单克隆抗体的制备及其初步应用

用CMV AD-169株感染自制人胚肺纤维母细胞，制备抗原，锣疲BALB／c小鼠，．取其脾细胞与Sp2／0-Ag14细胞融合，建立了一株稳定分泌抗HCMV特异性McAb的3B0杂交瘤细胞株。3B0McAb经鉴定属于IgG3亚娄。用间接免疫荧光法和间接酶免疫法证实为抗CMV特异性抗体。用微量细胞培养中和试验证实3B0McAb在加有豚鼠血清后可阻止HcMV感染后细胞病变的出现。3B0McAb经间接免疲荧光法检测中，晚期孕妇羊水细胞中HCMV抗原，在68例标本中有一例为阳性，阳性率为1．47％。该法方便，快速可为在胎儿时期诊断先天性HCMV感染提供依据。     

抗伏马菌素B_1单克隆抗体的制备及鉴定

目的建立抗伏马菌素B1（FB1）的单克隆杂交瘤细胞株,制备抗FB1的单克隆抗体（McAb）。方法采用FB1-KLH偶联物小剂量长周期免疫BALB/c小鼠后,采用细胞融合方法获得分泌抗FB1的杂交瘤细胞株,采用多次亚克隆的方法建立了4株稳定分泌抗FB1抗体的杂交瘤细胞株。腹水诱生法获得大量McAb,采用抗体亚类测定试剂盒鉴定抗体的亚类,SDS-PAGE测定抗体分子量,McAb的特异性和灵敏度用间接竞争抑制ELISA。结果免疫小鼠血清检测结果显示经过5次免疫后血清的效价稳定在1×10-6,经过4次亚克隆后建立4株稳定分泌抗FB1抗体的杂交瘤细胞株,获得腹水,纯化抗体,抗体亚类属于IgG2a类,轻链为κ,轻链和重链的相对分子质量分别是55000和32000,ELISA检测抗体特异性显示可与FB1发生特异性反应,采用此抗体建立间接竞争抑制ELISA方法的线性范围在2～500ng/ml。结论制备了特异性和灵敏度较高的抗FB1单克隆抗体,为建立伏马菌素免疫学检测方法打下良好基础。     

抗人IgA单克隆抗体的特性分析和初步应用

用常规免疫法和脾内直接注射抗原(SIgA)法免疫小鼠,平行作细胞融合实验,建立4株分泌抗人IgA McAb杂交瘤细胞株,所得McAb对人IgA有较高特异性和抗体效价,经免疫印染法表明对α重链呈特异性。用抗原竞争抑制ELISA和ELISA添加试验分析McAb的抗原结合位点,均表明McAb A4与A1b、A3b、A9针对不同的抗原位点。用于检测EB病毒VCA和EA的IgA抗体,显示满意的特异性和敏感性,这将为有关试剂的生产提供稳定的抗体来源。     

单克隆抗体亲和层析法纯化HFRS病毒血凝素抗原的初步研究

用蔗糖-丙酮法从HFRS病毒陈栋感染的乳小白鼠鼠脑中提取粗制血凝素,将此粗制血凝素通过抗HFRS病毒血凝素McAb 3G_1与Sepharose 4B偶联的免疫吸附柱,获得初步纯化的HFRS病毒血凝素抗原,经鉴定具有较高的血凝滴度和较好的免疫原性。用SDS-PAGE和Wes-tern-blot技木分析该纯化的血凝素抗原,并与同法纯化的正常鼠脑抗原比较,证明该特异性血凝素抗原分子量为5×10~4道尔顿。用2株抗HFRS病毒以及10株抗HFRS病毒血凝素McAb,以ELISA阻断试验分析纯化50K血凝素,结果有10株McAb可阻断McAb3 G_1与其结合,其抗原决定簇之间有部分相同或重叠,提示这些具有不同特性的抗原决定簇确实位于同一结构蛋白上。以上结果表明,该50k血凝素蛋白特性及结构均较复杂,尚待进一步研究。     

分泌抗A群脑膜炎球菌多糖单克隆抗体的杂交瘤细胞系的建立

用A群脑膜炎球菌悬液免疫的BALB/c小鼠脾细胞与Sp2/0骨髓瘤细胞,经50％PEG处理进行融合。采用微量被动血凝试验(PHA)筛选及有限稀释法克隆化后,获得4株分泌抗A群脑膜炎球菌多糖单克隆抗体(McAb)的杂交瘤细胞系(WSA—C_(11),WSA—E_6,WSA—D_4,WSA—F_8),培养上清和诱生的小鼠腹水的特异性抗体的PHA和ELISA滴度分别为1.28—5.12×10~(-2)和1×10~(-5)—10~(-6)、4株McAb和PHA滴度均被A群脑膜炎球菌多糖抗原抑制,其血凝抑制滴度与所加抑制抗原的量成线性关系,4株McAb均与3株A群脑膜炎球菌菌株呈现强的玻片凝集反应,3株B群,2株C群,1株D群,及7株新群脑膜炎球菌菌株不发生效义玻片凝集反应。试验证明4株杂交瘤细胞所分泌的抗体是针对具有群特异性的A群脑膜炎球菌多糖抗原的。4株McAb具有很高的亲和力,经80～160倍稀释与A群脑膜炎球菌菌株仍呈现强的玻片凝集反应,而A群脑膜炎球菌诊断血清(Pc-Ab)高于10倍稀与本菌的玻片凝集反应明显减弱,McAc的细菌试管定量凝集反应滴度也较PcAb诊断血清高16～32倍。4株McAb均属小鼠IgM类。杂交瘤细胞系染色体计数为91～95条。4株条交瘤细胞在体外连续培养4个多月,40余代,经液氮冻存复苏后仍保持稳定的分泌抗体的能力。     

抗人AFP单抗及多抗与丝裂霉素联接物对肝癌细胞的杀伤作用

用本室制备的分泌IgG AFP McAb的杂交瘤株G2及G10,接种于BALB/c小鼠腹腔,取其腹水经硫酸铵及Sepharose 4B柱亲和层析纯化,获得抗人AFP单抗。将马抗人AFP血清经硫酸铵及Sepharose 4B柱亲和层析提取AFP抗体IgG,得到AFP多抗。将纯化     

人胎盘层粘连蛋白单克隆抗体的制备与鉴定

目的获得能特异性识别人层粘连蛋白（hLN）的单克隆抗体（McAb）。方法用纯化的人层粘连蛋白（hLN）作抗原免疫Balb/C小鼠,以细胞融合、酶联免疫吸附实验（ELISA）筛选和克隆化技术获得抗hLN的杂交瘤细胞株;用生物学方法鉴定杂交瘤细胞,用免疫学方法鉴定McAb的特异性。结果获得4株稳定分泌抗人LN的杂交瘤细胞株（2A1、3B5、2C4、4D1）,培养上清的ELISA效价分别为：1∶512、1∶1024、1∶512、1∶256;腹水效价分别为：1×10^6、1×10^7、1×10^6、1×10^5;采用ELISA相加法表明2A1、4D1与3B5、2C4识别的hLN上的抗原决定簇和识别的不同,4株单抗均属实IgG。结论成功建立了4株稳定分泌抗人LNMcAb的杂交瘤细胞株,它们分别识别hLN上2个不同的抗原位点,有望作为hLN定量检测的特异性抗体。     

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.     

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.     

The universal muscular chain reaction,muscle spasm,Torsions, ruptures and extravasations. Chameleons of pathology and some manifestations of simple muscular disorders

Most bodily functions require the coordinated actions of complementary and supplementary paired muscle groups. Where this essential muscular cooperation is lacking, hollow organs may burst and others become literally screwed up, giving rise to many similar spastic diseases such as Torticollis, Twisted ovarian cyst, Torsion of the Testis, Volvulus of the intestines, Varicose Veins, Megacolon, Aortamegaly, Scoliosis, Erb's Palsy, Peyronie's Disease, Main-en-Griffe, Undescended Foot (Pes Cavus), Talipes, Strabismus. Spasm is “panenepidemic” and unclassified examples of Torsion Dystonia and Dyskinesia really are as common as debt and taxes.     

'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.     

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.     

Hypo- und Hypermagnesämie aus kardiologischer Sicht

Zusammenfassung Eine Reihe pathologischer Zustände bedingen Magnesiummangel. Zustände mit Hypermagnesämie sind ebenfalls bekannt, doch wesentlich seltener. Für den Kardiologen beachtenswert ist, daß unter Therapie mit bestimmten Diuretica bei Herzinsuffizienz, bei Herzinfarkt, Kardiomyopathie, Digitalisintoxikation und bestimmten Herzrhythmusstörungen Hypomagnesämie beobachtet wurde. Leider kann in der klinischen Routine nur ein extracelluläres Magnesiumdefizit durch Serumbestimmungen gemessen werden; über Magnesiummangel einzelner Organe kann nichts ausgesagt werden. Hinweise für Magnesiummangel geben aber neben der Messung des Serumspiegels Anamnese, klinischer Befund, bestimmte EKG-Veränderungen wie auch evtl. Hypokalämie, ein Zustand, bei dem sich oft — besonders bei Aldosteronismus — parallele Veränderungen zeigten.Tierexperimente deuten darauf hin, daß infarktähnliche Läsionen unter Magnesiummangel entstehen, doch ob Herzinfarkt beim Menschen durch Magnesiummangel ausgelöst werden kann, ist noch ungeklärt. In Leichenherzen zeigte sich im Infarktgebiet neben Calciumakkumulation signifikanter Magnesiumverlust, wobei unklar blieb, ob sich Ursache oder Folge des Infarktes widerspiegelten. Falls ein ursächlicher Zusammenhang besteht, ist er im Myokardstoffwechsel selbst zu suchen, wie bei der Alkoholkardiomyopathie, wo myokardialer Magnesiummangel zumindest als pathogenetischer Teilfaktor anerkannt wird. Andererseits versucht man aber auch Beziehungen zwischen Atherosklerose, Blutgerinnung und Hypomagnesämie herzustellen, in der Meinung, daß Magnesiummangel auch über den coronaren Pathomechanismus des Herzinfarktes wirken könnte. Sicher scheint, daß gewisse EKG-Veränderungen und Herzrhythmusstörungen durch einen irritierten Magnesiumhaushalt bedingt sein können, da sie bei Gabe bzw. Entzug von Magnesium verschwinden. Daß Magnesiummangel die Glykosidtoleranz verringert, wird tierexperimentell bestätigt. Unter Hypomagnesämie bewirkt Acetylstrophanthidin eher und länger Rhythmusstörungen als ohne, außerdem lassen diese sich durch Magnesiumgaben eliminieren. Da in gewissen Fällen spontane und digitalisinduzierte Herzrythmusstörungen durch Magnesiuminjektionen beseitigt wurden, scheint Magnesium als Therapeuticum angebracht. Einsatz verschiedener Magnesiumsalze bei Angina pectoris, degenerativen Herzerkrankungen und Herzinsuffizienz ohne geprüften und offensichtlich gestörten Magnesiumhaushalt ist fragwürdig, weil keine eindeutigen klinischen Erfolgsbeweise vorliegen. Immerhin mag es aber larvierte, durch Serumbestimmungen nicht erfaßbare Mangelzustände geben. Allgemein erscheint es aus kardiologischer Sicht ratsam, den Magnesiumhaushalt zu überwachen und in entsprechenden Fällen auszugleichen, um möglichen Myokardläsionen oder fatalen Herzrhythmusstörungen entgegenzuwirken.     

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.     

The case for allele‐specific recognition by the TCR

There is a sharp difference in how one views TCR structure–function–behaviour dependent on whether its recognition of major histocompatibility complex‐encoded restriction elements (R) is germline selected or somatically generated. The generally accepted or Standard model is built on the assumption that recognition of R is by the V regions of the αβ TCR, which is not driven by allele specificity, whereas the competing model posits that recognition of R is allele‐specific. The establishing of allele‐specific recognition of R by the TCR would rule out the Standard model and clear the road to a consideration of a competing construct, the Tritope model. Here, the case for allele‐specific recognition (germline selected) is detailed making it obvious that the Standard model is untenable.