卵巢上皮癌相关肿瘤抗原新基因的研究

目的探讨卵巢癌相关肿瘤抗原新基因在卵巢癌诊断和免疫治疗中的应用价值。方法利用网上生物信息学分析软件对筛选到的55个克隆的核苷酸序列进行分析,并检测其中8个新基因在卵巢癌患者和正常女性血清中的抗体。结果55个克隆的核苷酸序列代表45个卵巢癌抗原基因,其中8个是国内外首次发现的在GeneBank中尚未登记的新基因,是具有免疫原性的功能基因。其中OCY-142新基因是抑癌基因BARD1的可剪切变异体,可被CD4+和CD8+T细胞识别。8个卵巢癌肿瘤抗原新基因中7个在卵巢癌患者血清中的阳性率显著高于正常人,7个卵巢癌肿瘤抗原新基因对卵巢癌患者和正常女性进行联合检测,其敏感性和特异性达到96.9%和95.2%。CAl25异常的卵巢癌41例患者中,OCY-142单独检测的阳性率为95.12%,而CA125正常的卵巢癌57例患者中,OCY-142检测发现21例(36.9%)阳性。结论这些卵巢癌相关肿瘤抗原新基因在卵巢癌患者血清中产生的相应抗体可能成为卵巢癌的新的血清学标志物,可能是卵巢癌免疫治疗理想的新靶点。     

SEREX肿瘤抗原基因表达蛋白的研制和血清学初步筛选

目的 :利用基因工程方法获得SEREX肿瘤抗原基因原核表达蛋白 ,为进一步血清学分析奠定基础。方法 :采用SEREX方法 ,筛选中国人卵巢癌cDNA表达文库。对获得的 3个阳性克隆基因MY OVA 2、MY OVA 7和MY OVA 13,利用基因工程的方法克隆其全长基因 ,原核表达和亲和层析获得目的蛋白 ,并采用Dot blot的方法初步检测了目的蛋白抗原在正常人和肿瘤患者中的血清学反应格局。结果 :获得了纯化的目的蛋白 ,采用点杂交对 13例正常人和 74例肿瘤患者血清的测定结果显示 ,MY OVA 13的抗体只在肿瘤组中检测到 ,在正常组中为阴性 ;MY OVA 2和MY OVA 7在两组人群的血清中自身抗体检出率均呈现阳性 ,不同的肿瘤类型的阳性率各不相同。结论 :基因工程获得的蛋白质抗原可望成为肿瘤的血清学诊断工具     

人内质网分子伴侣BiP抗原表位筛选及其在RA血清学诊断中的作用研究

目的:筛选鉴定人内质网分子伴侣BiP抗原表位多肽,探讨其在类风湿性关节炎(Rheumatoid arthritis,RA)血清学诊断中的价值.方法:运用计算机软件分析BiP蛋白的结构特点及抗原决定簇分布,根据结果设计合成系列多肽.比较合成多肽与RA患者血清(BiP抗体阳性)IgG的反应强弱,鉴定抗原抗体反应最强的抗原表位多肽.进一步以获得的BiP抗原表位多肽作为包被抗原,检测79例RA患者、34例系统性红斑狼疮(SLE)患者、66例干燥综合征(SS)患者和173例正常人血清中抗BiP抗原表位多肽IgG抗体水平.结果:筛选到一条与RA患者血清反应最强的多肽(N403),抗N403多肽IgG抗体在RA患者中的阳性率(67%),明显高于SS患者(29%)、SLE患者(0%)、和正常人(0%)(P＜0.01).抗N403多肽IgG抗体在RA血清学诊断中的敏感性为67%,特异性为93%.而且该抗体的检测对于RF、CCP、HRF、RA33、AKA、APF等抗体阴性的RA患者的诊断具有重要意义.结论:筛选获得的BiP抗原表位多肽N403,其检测抗体在RA临床血清诊断中具有很好的应用价值.     

从卵巢癌cDNA文库中筛选新的肿瘤标志物

采用SEREX法筛选了自行构建的中国人卵巢癌cDNA表达文库 ,得到 2 7个阳性克隆 ,其中 3个为全长cDNA。序列分析和同源性比对结果表明所获得的 2 7个克隆分属于分化抗原、细胞结构蛋白及功能未知三类。选择其中一个全长cDNAMY OVA 13(该基因编码的蛋白是MAD2 ) ,利用基因工程方法克隆、表达和纯化得到目的蛋白 ,采用点杂交方法检测了MY OVA 13目的蛋白与正常人和肿瘤患者中的血清学反应。MY OVA 13抗体只在肿瘤组中检测到 (5 / 74 ) ,在正常组中均为阴性 ;间接ELISA测定结果显示 ,MY OVA 13抗体的水平在肿瘤组中均高于正常组 ,其中肝癌和前列腺癌组与正常组相比 ,在统计学上有显著差异 ,P值分别 <0 0 1和 <0 0 5。我们的初步结果表明MY OVA 13及其抗体具有一定的肿瘤特异性 ,有可能作为肿瘤诊断的标志物     

人肿瘤抗原分子分析的一种新技术

新近发展起的“血清学鉴定重组表达抗原克隆技术(serologicalidentificationoftumorantigensbyrecombinantexpressioncloning-SEREXapproach)”已成功在人恶性黑色素瘤,肾细胞癌,成胶质细胞瘤以及何杰金氏病,食道癌、肝细胞癌等证明多种新的肿瘤抗原,并从分子水平得到确定。本文介绍该技术的原理、程序。1.原理:利用病人自身血清含有肿瘤抗原的相应IgG抗体,筛选出自身肿瘤cDNA文库中与高滴度IgG抗体反应克隆,提交到PBK-CMV噬菌体内剪切后测定cDNA插入片段核苷酸的顺序。此法在致免疫的肿瘤蛋白与病人自身血清反应的基础上可直接确…     

血清CEA、CA125、CA724联检对卵巢癌的诊断价值

目的:探讨用肿瘤标志物癌胚抗原(CEA)、糖类抗原125(CA125)、糖类抗原724(CA724)联检对卵巢癌的临床评价。方法:采用电化学发光法测定28例卵巢癌及37例正常人血清。结果:以单一指标阳性作为诊断标准CEA、CA125、CA724对卵巢癌的敏感性分别为25.0%、42.8%、32.1%,特异性分别为91.4%、97.2%、86.5%,联检结果二项或二项以上阳性作为标准则本文患者的诊断灵敏度为78.6%、特异性为83.8%。结论:联检血清CEA、CA125与CA724对卵巢癌的辅助诊断有较高的临床应用价值。     

从卵巢癌cDNA表达文库筛选肿瘤抗原基因的研究

目的：利用肿瘤患者自体血清筛选患者肿瘤cDNA表达文库（SEREX）,寻找肿瘤特异性抗原是肿瘤免疫学研究的新策略,由此方法所获B细胞抗原肽可为进一步寻找T细胞抗原肽,开展肿瘤免疫治疗和诊断提供线索。方法采用SEREX方法,筛选中国人卵巢癌cDNA表达文库,对获得的阳性克隆片段测序鉴定和进行BLAST同源性比较。采用RT－PCR,分析政党组织和肿瘤组织及肿瘤细胞株中6个新的EST片段的表达。结果,所获基因片段大多数为已知抗原基因（20／27）。根据其来源又可归类为结构基因。线粒体基因和调控基因。另外,获6个未知的EST片段。RT－PCR的结果表明,4个EST（EST87888、1750、1754和3533）片 肿瘤和正常组织中均有表达,2个EST在正常组织中未见表达,而在肿瘤组织中有表达,可能是与肿瘤相关或特异的EST片段。结论SEREX方法不失为一种有效筛选卵巢癌肿瘤抗原的新方法。所获肿瘤抗原候选基因,可进一步进行结构和功能的研究。     

肝癌抗原特异性的检测

目的　检测两种新的人肝细胞癌 (HCC)抗原 (编号为HCC 1 8a和HCC 3 13)对HCC的特异性。方法　应用重组克隆表达抗原的血清学技术 (SEREX) ,用异体HCC血清和非HCC血清检测两种新的HCC抗原的特异性。结果　相应的抗体主要见于HCC患者 ,阳性反应率分别为 80 % (8/ 10 )及 90 % (9/ 10 ) ;胃癌为 0及 2 0 % (1/ 5 ) ;正常人各为 7.1% (2 / 2 8) ;乳癌为 0。结论　两种新的HCC抗原对HCC有高度特异性 ,可考虑作为HCC血清学诊断的指标之一。     

用自身血清抗体筛选的一种新的肝细胞癌表达的抗原

<正> 目的:寻找人肝细胞癌(HCC)特异抗原.方法:应用“重组子表达抗原克隆的血清学鉴定技术(SEREX)”,用病人自身血清从广西肝细胞癌cDNA文库中筛选出编码HCC肿瘤抗原的基因克隆,用肝癌患者及其他人血清检验HCC肿瘤抗原对肝癌的特异性,测定抗原DNA序列并与基因库核对寻找其同源结构.结果:其中一种HCC肿瘤抗原与衰老标记蛋白-30(SMP-30)是同源结构,相应抗体主要只见于HCC病人,在20例其它5种恶性肿瘤和4例肝硬化病人未查到相应抗体,20例慢性肝     

自身免疫性肝病患者自身抗体检测及临床意义

目的 探讨自身免疫性肝病患者血清中出现的自身抗体等免疫学指标及临床意义.方法 对3 500例肝功能反复异常的患者采用间接免疫荧光法检测抗核抗体(ANA)、抗平滑肌抗体(SMA)、抗线粒体抗体(AMA).并对AMAM2型及抗可溶性肝抗原/肝胰抗原(抗SLA/LP)、抗肝肾微粒体抗体Ⅰ型(抗LKM-1)和抗肝特异性胞浆抗原Ⅰ型抗体(抗LC-1)等肝脏疾病相关的自身抗体进行检测.结果 3 500例患者中,自身免疫性肝炎患者29例,检出率为0.83%,其中符合Ⅰ型、Ⅱ型、Ⅲ型自身免疫性肝炎的比例占72.4%、10.3%和17.2%.原发性胆汁性肝硬化(PBC)患者58例,检出率为1.65%,血清中AMAM2型抗体阳性率为93.1%,其中19例AMAM2阳性患者进行肝穿病理检查时12例(63.7%)患者病理提示符合PBC诊断.结论 每种自身免疫性肝病都具有特征性自身抗体谱,注重自身抗体检测对明确诊断及鉴别诊断自身免疫性肝病具有重要的临床意义.     

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 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.