gp130结合分子抑制JAK激酶与gp130的结合

ｇｐ１３０结合分子（ＧＡＭ）是以ｇｐ１３０胞浆内段为探针新近克隆成功的一个分子，其功能仍不清楚。为探讨ＧＡＭ在ｇｐ１３０信号传导中的可能作用，我们将不同长度的ｇｐ１３０变异体转染Ｈｅｐ３Ｂ细胞，观察ＧＡＭ与ｇｐ１３０的免疫共沉淀。结果表明ＧＡＭ可与ｇｐ１３０胞浆内近胞膜部分相结合。当ＧＡＭ、ｇｐ１３０和ＪＡＫ激酶共转染ＣＯＳ细胞时，ＧＡＭ的表达可明显抑制ＪＡＫ激酶对ｇｐ１３０的磷酸化。以上结果提示，ＧＡＭ可能作为一个抑制分子参与ｇｐ１３０的信号传导。     

GAMdDNA稳定转染的M1细胞系的建立与鉴定

目的 为搞清ＧＡＭ在ｇｐ１２３０介导的信号转导中所起的作用。方法 选择ＩＬ－６ 出现分化和增殖抑制的Ｍ１细胞系，分别用正向和反向插入ＧＡＭＣＤＮＡ的真核表达载体ＰＥＥ－ＢＯＳ与ｐＳＶ２－ｎｅｏ质粒共转染，经Ｇ４１８选择培养筛选得到稳定转染的细胞，并用半工ＰＣＲ方法产了ＧＡＭ基因与这两种细胞基因组ＤＮＡ的整合。结果 流式细胞仪分析表明正向ＧＭＡＣＤＮＡ转染的Ｍ１细胞中ＧＭＡ的表达水平明显升高在ｇｐ１     

gp130结合分子抑制JAK激酶与gp130的结合

ｇｐ１３０结合分子（ＧＡＭ）是以ｇｐ１３０胞浆内段为探讨新近克隆成功的一个分子，其功能仍不清楚。为探讨ＧＡＭ在ｇｐ１３０信号传导中的可能作用，我们将不几工度的ｇｐ１３０变异体传染Ｈｅｐ３Ｂ细胞，观察ＧＡＭ与ｇｐ１３０的免疫共沉淀，结果表明ＧＡＭ可与ｇｐ１３０胞浆内的膜部分相结合。当ＧＡＭ、ｇｐ１３０和ＪＡＫ激酶共转染ＣＯＳ细胞时，ＧＡＭ的表达可明显抑制ＪＡＫ激酶时ｇｐ１３０的磷酸化。以上结果提示：     

人树突状细胞cDNA质粒文库的构建及其大规模随机测序体系的 …

目的：建立人树突状细胞的ｃＤＮＡ文库。通过大规模随机测序克隆免疫新分子,方法：来源于正常人外周血的单核细胞,体外经ＧＭ－ＣＳＦ和ＩＬ－４培养,扩增出高纯度的ＤＣ,抽提纯化ｍＲＮＡ,转录成ｃＤＮＡ,定向插入ｐＳＰＯＲＴ２．０载体,建立人ＤＣ的ｃＤＮＡ质粒文库;用ＡＢＩ３７７全自动测序仪对该文库进行随机测序,将得到的ＥＳＴ在Ｓｕｎ－Ｅ４５０服务器中与ＥＭＢＬ及Ｓｗｉｓｓｐｒｏｔ数据库进行同源性比较,筛     

人补体C_1－抑制物基因在COS－7细胞中的表达

利用全长人Ｃ１－ＩＮＨｃＤＮＡ片段构建成功可在ＣＯＳ－７细胞中瞬间表达重组人Ｃ１－ＩＮＨ的表达质粒ｐＳＶＬＣ１。通过ＤＥＡＥ－Ｄｅｘｔｒａｎ法转染ＣＯＳ－７细胞，经体外标记、免疫沉淀和Ｗｅｂｔｅｒｎ印迹后发现，转染细胞可分泌─条分子量约１１０ｋＤ的免疫活性蛋白带。ＥＬＩＳＡ测定其表达水平，在转染后７２ｈ可达２μ／（ｍｌ·１０７细胞）。功能活性测定表明重组Ｃ１－ＩＮＨ具有抑制Ｃ１ｓ酯裂解的作用。     

可溶性syndecan-1 对人多发性骨髓瘤细胞体外的生物学作用

目的：研究可溶性ｓｙｎｄｅｃａｎ－１（ＣＤ１３８）分子对人多发性骨髓瘤（ＭＭ）细胞生长行为的影响及其机制。方法：用亲和层析法分离可溶性ｓｙｎｄｅｃａｎ－１;借助＾３Ｈ－ＴｄＲ掺入、间接免疫荧光、ＡＮＮＥＸＩＮ－Ｖ及ＰＩ标记分析ＭＭ细胞增殖、凋亡周期变化。结果：（１）从ＸＧ－２细胞培养上清中分离纯化得到分子量为６０ｋＤ左右的可溶性ｓｙｎｄｅｃａｎ－１分子;（２）可溶性ｓｙｎｄｅｃａｎ－１分子在体外能抑制ＸＧ－１增殖,阻滞细胞增殖停留在Ｇ２／Ｍ期。并介导其凋亡;ＦＧＦ、５％小牛血清、肝素酶Ⅲ可逆转一定浓度可溶性ｓｙｎ－ｄｅｃａｎ－１的作用,ＨＧＦ、ＶＥＧＦ和ＩＧＦ－１可部分逆转可溶性ｓｙｎｄｅｃａｎ－１分子对细胞的生长抑制作用;（３）ＩＬ－６及激发型抗ＩＬ－６Ｒ１３０（ｇｐ１３０）单抗对可溶性ｓｙｎｄｅｃａｎ－１ｗ     

中国版纳猪MHCI类P1分子全长的原核表达与纯化

目的：获得原核表达的中国版纳猪ＳＬＡＩ类Ｐ１蛋白质分子。方法：ＰＣＲ扩增去信号肽的ＳＬＡＩ类Ｐ１ｃＤＮＡ序列，亚克隆至ｐＧＥＭＴ载体，测序。将亚克隆的Ｐ１　ｃＤＮＡ片段插入表达载体ｐＥＴ４２ｂ（＋），构建重组表达质粒ｐＥＴ－４２ｂ（＋）／ｓｌａ－ｐｌ，转化Ｅ·ｃｏｌｉ表达菌 ＢＬ２１－ＣｏｄｏｎＰｌｕｓ（ＤＥ３）－ＲＩＬ，ＩＰＴＧ诱导 Ｐ１－８ ｘ ｈｉｓ融合蛋白表达，经包涵体洗涤，８ ｍｏｌ／Ｌ尿素变性溶解，Ｎｉ２＋亲和层析，梯度透析后，定量保存。ＳＤＳ－ＰＡＧＥ、ｗｅｓｔｅｒｎ－ｂｌｏｔｔｉｎｇ鉴定目的蛋白的表达与纯化。结果：目的蛋白（分子量３９．５ ｋＤ）表达量占细菌总蛋白 １５％，每升表达菌获得纯度９５％的目的蛋白 ４０ ｍｇ～６０ｍｇ。结论：成功建立猪 ＳＬＡ分子全长原核表达、纯化体系，为建立间接识别猪移植抗原ＳＬＡＩ类分子的人Ｔ细胞系及表位分析打下基础。     

人补体C1—抑制物基因在COS—7细胞中的表达

利用全长人Ｃ１－ＩＮＨ ｃＤＮＡ片段构成建功可在ＣＯＳ－７细胞中瞬间表达重组人Ｃ１－ＩＮＨ的表达质粒ｐＳＶＬＣ１。通过ＤＥＡＥ－Ｅｅｘｔｒａｎ法转染ＣＯＳ－７细胞，经体外标记、免疫沉淀和Ｗｅｓｔｅｒｎ印迹后发现，转染细胞可分泌一条分子量约１１０ｋＤ的免疫活性蛋白带。     

一种新的真核细胞表达载体pEF—BOS的应用

采用一种新的真核细胞表达载体ｐＥＦ－ＢＯＳ在ＣＯＳ细胞内表达了ｇｐ１３０和ＧＡＭ两种分子，并利用免疫沉淀和Ｗｅｓｔｅｒｎ－ｂｌｏｔ观察了表达结果，发现应用ｐＥＦ－ＢＯＳ表达载体可在ＣＯＳ细胞内获得上述两种分子的较高表达，优于一般常用的ｐＳＶ表达载体。     

人细胞间粘附分子1基因功能区Ⅰ,Ⅱ的克隆及高效表达

用ＰＣＲ聚合酶链反应技术从人细胞间粘附分子１（ＩＣＡＭ－１）ｃＤＮＡ中获取了其编码细胞外氨基端功能区Ⅰ、Ⅱ１８５个氨基酸的ＤＮＡ序列，构建了重组质粒ｐＥＴ／ｒｓＩＣＡＭ－１。经转化宿主菌，通过ＳＤＳ－ＰＡＧＥ，薄层扫描和Ｗｅｓｔｅｒｎ ｂｌｏｔ，证明目的基因在大肠杆菌中获得了高效表达。     

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.