骨碱性磷酸酶(BAP)酶联免疫吸附分析方法的建立及初步应用

目的 建立骨碱性磷酸酶（BAP）酶联免疫吸附分析方法（ELISA）.方法 使用两株抗体,一株包被微孔板,另外一株用HRP标记.显色系统使用TMB显色.结果 测定范围在5～ 120ng/mL,最低检测限在1.5ng/mL,回收率在96.4％～ 102％之间,批内和批间变异分别为5.6％和7.9％,正常参考值为＜10ng/mL,用502份临床样本考核本试剂盒,与参比试剂盒有很好的临床符合性,与参比试剂盒具有同等的临床使用价值.     

前列腺特异抗原化学发光免疫分析方法的建立及初步应用

建立的前列腺特异抗原(PSA)化学发光免疫分析(CLIA)使用2株抗PSAMcAb,一株McAb包被微孔板,另一株McAb与HRP连接。底物是鲁米诺/H2O2/对碘苯酚系统的双抗体夹心一步法,测定范围0.5～64ng/mL,最低检出值为0.13ng/mL,平均回收率为100.6%,批内和批间CV分别小于8.3%和11.5%,正常参考值小于3.5ng/mL。1000份血清临床考核,与参比试剂盒临床符合率一致,具有同等的临床诊断价值。     

血清铁蛋白时间分辨荧光免疫分析试剂盒的研制

目的 血清铁蛋白(sFER)时间分辨荧光免疫(TRFIA)分析试剂盒的研制.方法 采用双抗体夹心法建立sFER-TRFIA试剂盒,并对试剂盒的各项指标进行评价.结果 试剂盒的线性测量范围为2.2～1 070 ng/mL,灵敏度为0.22 ng/mL,批内、批间的精密度分别为4.6%～7.2%、5.4%～8.9%,与癌胚抗原(CEA)、甲胎蛋白(AFP)和人白蛋白无交叉反应.86份血清样本用本试剂盒与国外其他试剂盒同时检测,其相关系数为0.994.结论 试剂盒各项指标均达到临床检测要求,可替代国外同类产品试剂盒.     

糖类抗原CA72-4化学发光定量免疫分析方法的建立

目的 建立糖类抗原CA72-4化学发光定量免疫测定方法.方法 一株抗CA72-4单克隆抗体经微孔板固相包被与另一株酶标单克隆抗体建立双夹心反应体系,检测样品中CA72-4抗原的含量.通过与国外权威试剂盒血样测值的对比,分析该方法临床应用的可行性.结果 该分析方法线性范围为0～ 160 U/mL,灵敏度为0.55U/mL.批内CV值为:5.49％,4.53％;批间CV值为:8.12％,7.81％;回收率为:99.74％ ～ 104.11％;与CEA,CA125,CA19-9无交叉反应;与法国Cisbio analysis公司所生产的CA72-4免放药盒血清测值对比分析后,发现两者相关性较高,并确定了本分析方法的正常范围.结论 该分析方法的测定结果与参比试剂盒结果有较高符合率,可用于临床血清中CA72-4的检测.     

甘胆酸化学发光定量免疫分析方法的建立

目的 建立甘胆酸（CG）化学发光免疫检测方法.方法 采用竞争法,固相微孔L板包被技术,通过与参比试剂盒临床结果相关性的比较,保证其具有同等的临床诊断价值.结果 本试剂盒的线性范围为0.25～ 40 μg/mL,灵敏度为0.05μg/mL,批内精密度分别为7.1％和7.4％;批间精密度为11.3％和15.74％;样品的回收率在85％～115％之间.与透明质酸、Ⅳ型胶原、Ⅲ型前胶原氨端肽、层粘连蛋白、胆酸以及鹅脱氧胆酸的交叉反应率在0.010％～1.480％之间,均不会对CG的正常测定构成干扰.通过临床样品正常值的测定确定了正常值范围,与有关文献报道基本一致.同时,与北京北方生物技术研究所的放免药盒临床结果进行科学比较,两组结果的相关系数r为0.947.结论 该试剂盒与参比试剂盒的测定结果具有较高的符合率,应用于临床,可为慢性肝炎复发、轻度肝硬变、肝癌、肝胆胃肠消化系统疾病的研究诊断提供可靠的指标.同时,肝胆酸与血清Ⅳ型胶原、透明质酸和前白蛋白（PA）动态联检对于肝硬化的诊断、疗效及预后判断具有重要的临床意义.     

高灵敏均相酶免疫法测定血清皮质醇方法建立与性能评价

目的 建立高灵敏均相酶免疫法定量检测血清皮质醇.方法 对此方法的线性范围、准确度、精密度、抗干扰能力、临床可报告范围以及参考区间进行性能验证.结果 本研究建立的血清皮质醇均相酶免疫测定方法,线性范围为30.0～1200.0ng/mL,准确度相对偏差B≤10.0％,批内CV≤10.0％,批间差R≤10.0％,当样本中胆红素≤50mg/L、血红蛋白≤1000mg/L、白蛋白≤100g/L、抗坏血酸≤176mg/dL时,对测定结果无影响,临床可报告范围上限6500ng/mL,血清皮质醇参考区间为60～230ng/mL.结论 该均相酶免疫法能够满足临床血清皮质醇检测的需求,可以进一步推广至临床使用.     

化学发光免疫分析血清C肽方法建立

双抗体夹心一步法建立人C肽(C-peptide) 化学发光免疫分析(CLIA)方法.方法可测范围0.15～15ng/mL,灵敏度0.02ng/mL,批内、批间变异系数(CV)分别为4.2%～6.5%和6.8%～9.3%,与胰岛素无交叉反应,与胰岛素原的交叉反应率为7.3%.本方法与国外同类试剂盒同时检测40份C-肽释放试验血清,相关系数为0.9709.本方法各项指标均满足临床检测要求,达到国外同类产品水平.     

不同抗干扰能力STA-Liatest D-Di试剂盒检测类风湿患者D-二聚体水平效果比较研究

目的 分析比较不同抗干扰能力STA-Liatest D-Di试剂盒检测类风湿患者D-二聚体水平的效果.方法 本研究选取本院就诊的确诊类风湿患者104例,使用STA-Liatest D-Di试剂盒进行DD120CN的检测,并与标准免疫比浊法检测结果进行比较,同时对两种试剂盒的灵敏度与特异性进行比较和评价.结果 本研究中,纳入研究的受试对象包含女性80例,男性24例,对于RF水平小于50 IU/mL的49例患者进行检测,STA-Liatest D-Di 1000IU/mL试剂盒检测的D-二聚体水平为8.59±1.16 ng/mL,STA-Liatest D-Di 50IU/mL试剂盒检测结果为8.65±1.21ng/mL,两种试剂盒间差异无统计学意义.对于RF水平大于50 IU/mL的55例患者进行检测,STA-Latest D-Di 50IU/mL试剂盒检测结果更高,P＜0.01.在女性受试对象中STA-Liatest D-Di 1000IU/mL试剂盒检测的特异度更高,P＜0.05.在男性受试对象中,两种试剂盒检测的灵敏度均为90.00％,STA-Latest D-Di 1000IU/mL试剂盒检测的特异性更高,P＜0.01.结论 检测工作中,对于RF水平大于50 IU/mL的患者来说STA-Liatest D-Di 50IU/mL更显优势.且无论对于男性还是女性患者,STA-Liatest D-Di 50IU/mL试剂盒和STA-Liatest D-Di 1000 IU/mL试剂盒对于检测风湿患者D-二聚体水平的灵敏度均较高;STA-Liatest D-Di 1000 IU/mL试剂盒的检测特异性则稍高于STA-Latest D-Di 50IU/mL试剂盒.     

促甲状腺素吖啶酯化学发光免疫分析方法的建立

目的 建立一种定量检测人血清促甲状腺素(TSH)的方法.方法 根据双抗体夹心法实验原理,分别用吖啶酯和生物素标记两株不同的单克隆抗体,链霉亲和素包被微粒,通过链霉亲和素-生物素系统分离固相和液相完成检测.结果 本方法的最低检测限为0.01 μIU/mL,线性范围0.3～60μIU/mL,批内变异为1.9％ ～2.3％,总变异为6.4％ ～8.6％,与ROCHE公司的促甲状腺素检测试剂盒(电化学发光法)比对,相关系数r=0.9935.结果 本方法的各项指标均能满足临床检测的要求,可以临床推广应用.     

4种不同方法学对可溶性生长刺激基因2蛋白试剂盒性能的初步评价

目的 4种不同方法学对可溶性生长刺激基因2蛋白(sST2)检测试剂盒的性能进行评价,并以ELISA为参比方法进行方法学比对。方法 收集2021年1月至3月首都医科大学附属北京安贞医院42名不同疾病的住院患者和健康体检中心20名表观健康人临床检测剩余血清。参考美国临床和CLSI文件和行业标准要求,对POCT免疫层析方法试剂盒(简称A)、增强免疫荧光方法试剂盒(简称B)、乳胶免疫比浊方法试剂盒(简称C)和磁微粒化学发光方法试剂盒(简称D)进行精密度、正确度、线性范围、参考区间、抗干扰能力、可报告范围方面的性能评价,以ELISA经典试剂盒作为参比方法(简称P),进行方法学比对和医学决定水平处偏差的计算。结果 A试剂的批间CV>15%,A和C试剂的平均回收率>20%,类风湿因子(RF)对C试剂干扰CV>10%;A试剂与参比方法试剂检测结果决定系数R²<0.90;4种试剂的可报告范围均>8 000.00ng/mL;4种试剂和参比试剂在医学决定水平处偏差均>10%。结论 A试剂(POCT免疫层析方法)和C试剂(乳胶免疫比浊方法试剂盒)性能评...     

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.     

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.     

What will studies of Fulani individuals naturally exposed to malaria teach us about protective immunity to malaria?

There are an estimated over 200 million yearly cases of malaria worldwide. Despite concerted international effort to combat the disease, it still causes approximately half a million deaths every year, the majority of which are young children with Plasmodium falciparum infection in sub-Saharan Africa. Successes are largely attributed to malaria prevention strategies, such as insecticide-treated mosquito nets and indoor spraying, as well as improved access to existing treatments. One important hurdle to new approaches for the treatment and prevention of malaria is our limited understanding of the biology of Plasmodium infection and its complex interaction with the immune system of its human host. Therefore, the elimination of malaria in Africa not only relies on existing tools to reduce malaria burden, but also requires fundamental research to develop innovative approaches. Here, we summarize our discoveries from investigations of ethnic groups of West Africa who have different susceptibility to malaria.     

'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.     

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.