结核杆菌Ag85B DNA疫苗的制备及其免疫效应初探

将结核杆菌抗原Ａｇ85Ｂ基因克隆到真核表达质粒ｐｃＤＮＡ3中 ,构建成重组ｐｃＤＮＡ3 Ａｇ85Ｂ ,作为抗结核的ＤＮＡ疫苗 ,观察其对小鼠免疫应答的能力 ,为进一步研制抗结核疫苗提供依据。1 结核杆菌抗原Ａｇ85Ｂ真核表达载体的构建及在Ｃｏｓ 7细胞中的瞬时表达 :由本室构建的重组质粒ｐＵＣ1 8 Ａｇ85Ｂ ,经ＥｃｏＲⅠ /ＳａｌⅠ酶切 ,获得 860ｂｐＤＮＡ片段 ,插入经ＥｃｏＲⅠ /ＸｈｏⅠ酶切的载体ｐｃＤＮＡ3中 ,用ＨｉｎｄⅢ和ＸｈａⅠ酶切分析鉴定阳性克隆 ,获得表达载体ｐｃＤＮＡ3 Ａｇ85Ｂ。用 5μｇ纯化的重组质粒ｐｃＤＮ…     

中国莱姆病螺旋体rRNAA基因多态性分析

本文作者首次应用地高辛标记的大肠杆菌１６＋２３ＳｒＲＮＡ基因探针分析中国莱姆病螺旋体ｒＲＮＡ基因限制性图谱。４５株中国菌株和６株国外菌株用于实验，选用ＥｃｏＲⅤ和ＨｉｎｄⅢ两种限制性内切酶。实验结果表明，中国菌株遗传差异性较大，分属三个基因种，即Ｇｅｎｏｓｐｅｃｉｅｓ Ⅰ，ＧｅｎｏｓｐｅｃｉｅｓⅡ和ＧｅｎｏｓｐｅｃｉｅｓⅢ，其中Ｂ．ｂｕｒｇｄｏｒ ｆｅｒｉ Ｓｅｎｓｕ Ｓｔｒｉｃｔｏ基因种在亚洲菌     

问号赖型钩端螺旋体重组质粒pDL121的初步研究

钩端螺旋体病的传染和流行常给人类生命及畜牧业生产造成严重危害。我国曾发生过数十次大规模钩端螺旋体病流行。作为我国两个流行菌型之一的问号赖型钩端螺旋体在生物学特性和致病性有其独特性，但其遗传学特性和致病机制尚不完全清楚。为了探讨问号赖型钩端螺旋体抗原基因的特点，应用六种常见内切酶（ＢａｍＨＩ，ＢｇｌⅡ，ＥｃｏＲＩ，ＨｉｎｄⅢ，ＰｓｔⅠ，ＰｖｕⅡ）对从问号赖型钩端螺旋体０１７株的基因文库中筛选出来的重组质粒ｐＤＬ１２１外源基因进行酶谱分析，同时用Ｄｉｇｏｘｉｎ标记的ｐＤＬ１２１外源基因片段作探针对不…     

人B细胞激活抗原B7（hB7）基因cDNA的克隆

我们采用Ｎｅｓｔｅｄ ＰＣＲ的方法，成功地从Ｒａｊｉ细胞中扩增出Ｂ７ ｃＤＮＡ，通过单个碱基改变，在两端分别引入单酶切位点（５＇端为ＨｉｎｄⅢ，３＇端为ＥｃｏＲⅠ），经酶切后连入ｐＢｌｕｅｓｃｒｉｐｔ克隆载体，测序证明所得序列与Ｆｒｅｅｍａｎ等报告的完全一致。我们所选用的两个酶切位点ＨｉｎｄⅢ和ＥｃｏｒＩ恰好位于ｐＢｌｕｅｓｃｒｉｐｔ多克隆位点的中央，克隆入Ｂｌｕｅｓｃｒｉｐｔ后，两端有多个单酶切     

我国登革2型病毒E基因的克隆与表达研究

以我国登革２型病毒４３（Ｄ２－４３）株ＲＮＡ为模板，采用ＲＴ－ＰＣＲ技术扩增了Ｅ基因。扩增的ｃＤＮＡ片段约１２９０ｂＰ， 与预期大小相一致，经ＨｉｎｄⅢ酶切鉴定和Ｓｏｕｔｈｅｒｎ印迹杂交证实了Ｅ基因片段的特异性。将Ｅ基因的扩增片段首先克隆到ｐＢｌｕｅｓｃｒｉｐtⅡＫＳ~＋质粒ＤＮＡ中，利用重组子中载体的酶切位点，将该基因片段再克隆到高效表达载体ｐＢＶ２２０中。用ＰＣＲ技术从２０个重组子中筛选出６个阳性克隆　，经ＳａｌⅠ和ＰｓｔⅠ酶切进一步鉴定，证明这６个重组质粒中均含有Ｅ基因片段。采用温控诱导，４小时表达的蛋白量约占菌体总蛋白的２０％。经Ｗｅｓｔｅｒｎｂｌｏｔ分析证明，表达的蛋白可与该病毒株的多克隆腹水抗体和登革２型病毒标准株的单克隆抗体起特异反应。     

中外七个贾第虫株DNA限制性酶切片段长度多态性（RFLP）比较研究

本文首次对分离自中国、美国、柬埔寨和澳大利亚７株贾第虫基因组ＤＮＡ，用６种限制性内切酶消化并进行琼脂糖电泳，分析比较它们基因组ＤＮＡ酶切片段多态性；同时用生物素标记的贾第虫全基因组ＤＮＡ探针对国内４株贾第虫ＤＮＡ进行Ｓｏｕｔｈｅｒｎ印迹杂交。结果表明，７株贾第虫ＤＮＡ经ＨｉｎｄⅢ和ＢｇｌⅡ酶切后，可将国内４株和国外３株贾第虫区别开；国内４株贾第虫ＨｉｎｄⅢ、ＢｇｌⅢ、ＨａｅⅢ、ＰｓｔⅠ和Ｈｉｎｆ５种内切酶酶切图谱一致；而经ＡｌｕＩ酶切后，Ｃ１和ＣＲ的ＤＮＡ图谱一致，Ｃ２和Ｃ３的也一致；Ｓｏｕｔｈｅｒｎ印迹杂交显示，国内４株间的ＤＮＡ杂交带相似，提示分离自人和兔的贾第虫无严格的宿主专一性，二者可产生交叉感染；国内３株与国外２株人贾第虫ＤＮＡ图谱存在差异，其原因似与地理隔离有关。     

HBV亚片段基因探针的制备及其在肝癌研究中的应用

以ＥｃｏＲⅠ酶切质粒ｐＢＲ３２２－２ＨＢＶＤＮＡ，回收ＨＢＶ３．２ｋｂＤＮＡ，继以ＢａｍＨⅠ、ＢｇｌⅡ酶切ＨＢＶ－ＤＮＡ，回收ＨＢｓ０．９ｋｂ及ＨＢｘ０．５８ｋｂＤＮＡ片段。以地高辛随机引物法标记上述回收片段，获得高度敏感性及特异性的ＨＢＶ全基因及亚片段基因探针。以ＨＢＶ全基因探针通过斑点杂交、Ｓｏｕｔｈｅｒｎ转膜杂交检测了４４例肝细胞性肝癌组织标本中ＨＥＶ－ＤＮＡ的存在情况及存在特点，继以ＨＢｓ、ＨＢｘ亚基因探针检测了ＨＥＶ纯整合型肛癌标本中各亚基因片段的整合情况。结果显示：７５％肝癌组织中有…     

利用杆状病毒载体在昆虫细胞中表达人白细胞介素2

利用杆状病毒载体在昆虫细胞中表达人白细胞介素２金立杰刘东升薛伟钢将人白细胞介素２（ＩＬ－２）ｃＤＮＡ在ＢａｍＨⅠ－ＰｓｔⅠ位点插入到苜蓿银纹夜蛾核型多角体病毒（ＡｃＮＰＶ）的转移载体ｐＶＬ１３９２中。经ＢａｍＨⅠ、ＰｓｔⅠ、ＸｂａⅠ、ＥｃｏＲⅤ等限制...     

中国莱姆病螺旋体rRNA基因多态性分析

本文作者首次应用地高辛标记的大肠杆菌１６＋２３ＳｒＲＮＡ基因探针分析中国莱姆病螺旋体ｒＲＮＡ基因限制性图谱。４５株中国菌株和６株国外菌株（美国Ｂ３１，２９７；法国２００４７；瑞士ＶＳ４６１；德国ＬＷ２和俄罗斯菌株ＩＰ２ｌ）用于实验，选用ＥｃｏＲⅤ和ＨｉｎｄⅢ两种限制性内切酶。实验结果表明，中国菌株遗传差异性较大，分属三个基因种，即ＧｅｎｏｓｐｅｃｉｅｓⅠ（Ｂ．ｂｕｒｇｄｏｒｆｅｒｉＳｅｎｓｕＳｔｒｉｃｔｏ），ＧｅｎｏｓｐｅｃｉｅｓⅡ（Ｂ．ｇａｒｉｎｉ）和ＧｅｎｏｓｐｅｃｉｅｓⅢ（Ｂ．ａｆｚｅｌｉ），其中Ｂ．ｂｕｒｇｄｏｒｆｅｒｉＳｅｎｓｕＳｔｒｉｃｔｏ基因种在亚洲菌株中是首次发现。此外还发现中国菌株ＨＢ１８的ｒＲＮＡ基因杂交图谱与国内外其它菌株不同。分属于三个基因种的中国菌株的ｒＲＮＡ基因杂交图谱与欧洲和美国菌株略有差异。中国菌株ＨＢ１８可能属于一个新的基因种。     

大肠杆菌菌株（λ衍生噬菌体）／PBR322衍生质粒作为人IL—6表 …

为探索人白细胞介素－６在原核细胞中高效表达的途径，选用ＢＬ２１／ＰＥＴ－２８作为表达系统，以聚质反应技术扩增了ｈＩＬ－６的经补ＤＮＡ的编码区，引入了ＥｃｏＲＩ和ＨｉｎｄⅢ位点。用限制性内切酶ＥｃｏＲＩ和ＨｉｎｄⅢ酶切后，克隆于ＰＥＴ－２８载体的相应位点，转化ＤＨ５α。选出必克隆，经ＤＮＡ序列分析，表明其编码区内无非特异突变。将重组质粒转子ＤＨ５α，筛选阳性克隆，用异基硫代β－Ｄ－半乳糖苷进行诱导表     

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.