肝癌相关抗原HAg18的研究及其应用

用抗人肝癌单克隆抗体HAb18亲和层析纯化相应抗原,经SDS-PAGE及免疫印迹试验证实其分子量为61kD。抗原活性条带经高碘酸钠氧化及氯仿-甲醇-冰乙酸处理抗原无丢失,但经胰蛋白酶酶解后失活。PAS及苏丹Ⅲ染色均为阴性。用HAb18ELISA双抗体夹心法可检测肝癌病人血清中抗原水平,阳性率为65.1％(250/384)。HAb18配伍另一肝癌单克隆抗体,制成ELA快速诊断盒,肝癌检出率提高至70.6％(271/384)。免疫酶斑点测定HAg18与AFP、铁蛋白、C-EA无交叉。故认为HAb18是一新的肝癌蛋白性抗原。     

转铁蛋白受体在肝癌细胞上的表达

作者用抗转铁蛋白受体单克隆抗体抗TfR McAb,(D71)检测17例原发性肝癌患者肿瘤组织。发现14例肝癌细胞呈现特异性结合,阳性率为82.4％。其余3例肝癌细胞未检测到转铁蛋白受体(TfR),可能与肝癌细胞的分化程度或分泌程度有关。同时用抗A-FP单克隆抗体与CD71进行比较,发现肝癌胞细TfR含量高于A-FP含量。该发现为肝癌的诊断及导向治疗提供了新的较好的载体。     

肝癌单克隆抗体-柔红霉素结合物的制备及其选择性细胞毒作用的研究

我室研制的HAb18单克隆抗体特异性高,免疫组化肝癌阳性率80％,在荷肝癌裸鼠及肝癌病人体内显像均良好。以HAb18与柔红霉素(DAU)连续制备了HAb18-Dextran-DAU免疫抗癌药物。经检测,H-Ab18-Dex-DAU结合物中抗体仍保持其特异性结合能力,证明制备过程中抗体活性未受到明显损伤;48h细胞毒性试验显示结合     

人肝细胞癌单克隆抗体的免疫组化定位及鉴别诊断的初步应用

用临床手术切除的新鲜肝癌组织作为抗原免疫动物,细胞融合,筛选出一株抗人肝癌细胞膜的单克隆抗体。此抗体采用ABC法对50例临床病理诊断为肝细胞肝癌的组织切片作免疫组化定位诊断,结果44例为阳性,6例为阴性,阳性率为89％。但有两例阴性在电镜观察,一例见到神经分泌颗粒,确诊为胰腺癌。另一例确诊鳞癌。抗体对35例肝外肿瘤的定位表明,     

抗人HAb18G分子单链抗体基因的构建及在大肠杆菌中的分泌表达

目的 :构建抗人HAb18G分子单链抗体基因 ,并在大肠杆菌中进行分泌表达。方法 :通过连接肽 (Gly4Ser) 3基因序列将已成功克隆的抗人肝癌单抗HAb18轻、重链可变区基因拼接成单链抗体 (ScFv)基因 ,并在 5′和 3′端引入相应的酶切位点 ,克隆至改造的分泌性表达载体pCANTAB 5His之中 ,转化到E coliHB2 15 1中进行诱导表达。亲和层析纯化表达蛋白后 ,以SDS PAGE电泳、Westernblot及ELISA等方法分析检测表达产物。结果 :经限制性酶切鉴定及DNA测序分析 ,证实基因构建正确。SDS PAGE电泳和Westernblot分析表明ScFv基因在大肠杆菌中成功表达。表达产物的相对分子质量 (Mr)为 31kD ,与理论预期值相符 ,并且可与相应的抗原特异结合。结论 :成功实现了抗人HAb18G分子单链抗体的原核分泌表达 ,为其在人肝癌诊治中的进一步应用奠定了基础。     

细胞外基质蛋白1在肝癌中的表达及意义

目的：探讨细胞外基质蛋白1（extracellular matrix protein1,ECMl）在肝癌中的表达及意义。方法：应用免疫组织化学EnVision法,检测肝脏细胞株（正常肝细胞株LO2、肝癌细胞株HepG2）和肝脏组织（正常肝组织和肝癌组织）中ECM1的表达,并比较正常肝组织和肝癌组织之间的ECM1表达差异。结果：ECM1的表达如下：正常肝细胞株LO2（-）,肝癌细胞株HepG2（＋）;正常肝组织阳性率20％（4／20）,肝癌组织阳性率85．4％（70／82）。统计分析表明,对ECM1的表达,肝癌组织明显高于正常组织（P〈0．01）。结论：ECM1在肝癌组织中过表达。     

抗人肝癌单克隆抗体HL2的制备及鉴定

用肝癌细胞株QGY-7703、BEL-7402、SMMC-7721以贯序法免疫BALB/c小鼠,获得一株分泌肝癌单抗的杂交窟株HL_2,其染色体数目为97—105。单抗HL_2系IgG_2b亚类。吸收实验及阻断实验证明HL_2抗原与CEA、AFP、HBsAg、HBcAg及HBeAg无免疫同源性。ABC法和ELISA法说明单抗HL_2与四株肝癌细胞、六例肝癌组织均呈明确阳性反应,除与SGC-7901、H_(128)、K_(562)、Hela细胞株及部分胚胎肝脏,胚胎结肠有较弱性反应外,与肝癌旁组织、正常肝脏、其它肿瘤组织和细胞株、二种正常人细胞及其它胚胎组织无反应。显示了单抗HL_2的特异性较好、阳性率较高,是一个有应用前景的单抗。     

抗人双特异性蛋白磷酸化酶Dusp23单克隆抗体的制备及鉴定

目的制备高亲和力、高特异性的抗人双特异性磷酸化酶23（Dusp23）单克隆抗体,并对其生物学特性进行鉴定,为Dusp23功能的研究奠定基础。方法以纯化的原核表达Dusp23为免疫原,免疫BALB／c小鼠。待免疫小鼠血清效价满足融合需要,取其脾细胞与Sp2／0细胞融合,经多次筛选及克隆化建立可稳定分泌抗Dusp23单克隆抗体的杂交瘤细胞株,将细胞株打入BALB／c小鼠腹腔制备腹水,用ProteinG亲合层析柱纯化所得腹水获得纯化抗体,ELISA及Western—blot检测抗体的效价和亲和力,并用亚型鉴定试纸条鉴定单克隆抗体的亚型。利用纯化后的两株单克隆抗体对临床收集的肝癌组织标本进行免疫组化分析。结果筛选到2株（N012和N013）可以稳定分泌人双特异性磷酸化酶23（Dusp23）单克隆抗体的细胞株,并对其进行纯化和鉴定,两株杂交瘤细胞培养上清效价分别为1：5120和1：2560,腹水效价分别为1：25600和1：12800,以肝癌细胞HepG2和重组Dusp23蛋白为抗原,利用纯化后的两株抗体Western-blot鉴定结果均在预期位置出现阳性条带。两株杂交瘤细胞分泌抗体的亚型鉴定N012和N013均为IgG1型,轻链均为K型。两株抗体分别与临床肝癌组织标本免疫组化结果均为阳性。结论成功制备两株能特异性识别Dusp23的单克隆抗体,为进一步研究Dusp23的生物学功能,揭示其与肿瘤发生发展之间的关系奠定了基础。     

抗人胶原蛋白单克隆抗体Ab15、Ab16的研制

用新鲜肝癌标本细胞悬液免疫研制抗肝癌单克隆抗体时,获得二株分泌抗人胶原蛋白抗体的杂交瘤细胞株Ab15、Ab16,并用此抗体在部分组织进行了免疫组织化学定位,初步确定为抗人Ⅳ型胶原蛋白,认为这二株抗体在Ⅳ胶原蛋白的免疫病理研究中将有一定的使用价值。     

人肝细胞癌单克隆抗体的免疫组化定位及鉴别诊断的初步应用

用临床手术切除的新鲜肝癌组织作为抗原免疫动物,细胞融合,筛选出一株抗人肝癌细胞膜的单克隆抗体。此抗体采用ABC法对50例临床病理诊断为肝细胞肝癌的组织切片作免疫组化定位诊断,结果44例为阳性,6例为阴性,阳性率为88％。但有两例阴性在电镜观察,一例见到神经分泌颗粒,确诊为胰腺肿瘤。另一例确诊鳞癌。抗体对35例肝外肿瘤的定位表明,仅对少数鳞癌和移行细胞癌有交叉反应。与全身绝大部份脏器组织无交叉反应。因此认为,此抗体对肝癌组织有一定特异性,对肝癌细胞亲合力较好,对临床活检诊断与鉴别诊断有潜在的实用价值。     

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.     

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.