质谱技术分析多发性骨髓瘤患者血清差异蛋白

目的应用双向电泳(2-DE)和质谱技术分析多发性骨髓瘤(MM)患者血清中的差异表达蛋白,寻找MM特异性蛋白标记物。方法收集MM患者5例、其他血液系统恶性肿瘤患者及正常对照者血清各10例,三组样本等体积混合,去除高丰度白蛋白和IgG,应用2-DE分离3组血清样本,凝胶经银染显色和图像数据分析识别差异蛋白质点,以基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)对差异蛋白质点进行鉴定。结果通过凝胶软件分析MM与疾病对照组及正常对照组血清2-DE图谱,共得到51个三倍以上差异蛋白点,对上述差异蛋白进行MALDI-TOF-MS质谱分析,共鉴定出22种阳性蛋白,其中13种蛋白表达上调,包括高表达蛋白如丝氨酸/苏氨酸蛋白激酶RIO1亚型2、Rab相关蛋白Rab37亚型2、HP蛋白、锌指结构α2糖蛋白等;9种蛋白表达下调,包括α2-HS-糖蛋白、转铁蛋白、多聚素-1等。结论 MM组与疾病对照组和正常对照组间存在差异表达的蛋白质,这些差异蛋白有助于MM早期诊断和鉴别诊断,并为MM发病机制的进一步研究提供线索。     

中医证侯血浆蛋白质双向凝胶电泳方法的建立及优化

目的建立中医证侯血浆蛋白质双向凝胶电泳（2-DE）方法并对其进行优化。方法以正常人血浆样品为研究对象,对白蛋白/IgG球蛋白去除后血浆样品、全血浆和热SDS法全血浆样品进行pH梯度2-DE分离,凝胶经硝酸银染色后,利用PDQuest软件对2-DE图谱进行重复性分析,并对不同处理方法的全血浆2-DE图像的结果进行比较。结果与全血浆直接上样2-DE凝胶图谱比较,白蛋白/IgG球蛋白去除后血浆上样双向电泳蛋白有所增多,但由于白蛋白/IgG球蛋白去除过程中的非特异性吸附,造成了同一样品在蛋白质图谱上的明显差异;与全血浆直接上样和白蛋白/IgG球蛋白去除后血浆样品上样2-DE凝胶图谱比较,热SDS法处理血浆样品可以得到更多的蛋白质,且重复性好（P〈0.05）,凝胶硝酸银染色显示蛋白质点染色深、分散清楚。结论热SDS法全血浆上样2-DE分析法具有蛋白丢失少、操作简便、重复性好的特性,是证侯血浆蛋白质组学研究的有效手段。     

HCV患者外周血单个核细胞蛋白质表达质谱分析

目的:应用比较蛋白质组学的方法分析丙型肝炎患者外周血单个核细胞蛋白质表达模式的变化及寻找丙型肝炎相关生物标记分子,进一步识别鉴定其差异表达蛋白质,分析其对丙型肝炎慢性化机制的意义.方法:应用固相化pH梯度双向凝胶电泳(2-DE)分离健康者(10)及HCV患者(28)PBMC的总蛋白质,凝胶银染显色后,PDQuest图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)获得差异蛋白点的肽质指纹图谱,通过SWISS-PROT数据库鉴定蛋白质.结果:得到两张2-DE图谱,HCV患者及健康者PBMC凝胶的蛋白质点数分别为625及614;初步筛选出HCV患者与健康者存在明显差异的12个蛋白点,经质谱分析,初步鉴定了10种蛋白质.这些差异蛋白质包括病毒蛋白、蛋白质合成与分解、三大代谢相关酶类、细胞结构相关蛋白质以及信号转导相关蛋白质.结论:应用2-DE及MALDI-TOF-MS方法建立了HCV患者PBMC双向凝胶电泳图谱,分离并初步鉴定了10种与HCV感染相关的差异表达的蛋白质,为研究HCV慢性化相关机制提供新的线索.     

脑脊液蛋白质组学高丰度蛋白去除方法的建立与评价

目的 建立和评价脑脊液蛋白质组学中高丰度蛋白的去除方法.方法 使用Oasis HLB(hydrophilic-lipophilic balance)过滤柱,运用反相固相法,根据蛋白质不同的疏水性分馏脑脊液,去除高丰度蛋白和盐.将含有低丰度蛋白的过滤液冻干,利用双向凝胶电泳(2-DE)分析评价脑脊液蛋白层析去除高丰度蛋白的效果.结果 通过Oasis HLB柱处理可将大部分高丰度蛋白去除.2-DE图谱中低丰度蛋白质分辨率明显增加,点数增多59.86%.结论 本研究建立的脑脊液高丰度蛋白去除方法可有效地应用于疾病的脑脊液蛋白质组学研究.     

H22肝癌小鼠血清差异蛋白质的质谱鉴定

目的 分析正常小鼠和H22肝癌小鼠血清之间差异表达的蛋白并对部分差异蛋白进行鉴定分析,筛选可能与肝癌发生密切相关的蛋白质.方法 利用双向凝胶电泳技术（2-DE）与基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）,分析正常组和肿瘤组筛选并鉴定部分差异表达的蛋白,再进行生物学分析.结果 通过2-DE图谱研究发现,与正常组相比,肿瘤组存在明显差异的蛋白质点.选择其中可能为关键功能蛋白的蛋白点进行质谱分析,获得肽指纹图谱（PMF）,运用Mascot软件查询NCBI数据库分析并鉴定得出其中2个蛋白：假尿苷合成酶和线粒体核糖体大亚基蛋白.结论 肝癌的发生机制与假尿苷合成酶和线粒体核糖体大亚基蛋白2个差异表达蛋白密切相关,其参与肝癌的发生发展.     

老年性痴呆大鼠血清的蛋白质组学研究

目的以老年性痴呆（AD）大鼠为研究对象,应用蛋白质组学方法在血清中筛选与AD密切相关的蛋白质,为研究AD的发病机制提供理论支持。方法通过Morris水迷宫记忆行为学筛选符合痴呆标准的AD大鼠模型,收集AD组大鼠及假手术组大鼠的血清后,去除血清中的高丰度蛋白质并进行蛋白质浓度的测定,之后进行双向凝胶电泳,挑选差异最为明显的蛋白质点进行肽质量指纹图谱分析,经数据库查询,鉴定差异蛋白质。结果经观察二维凝胶电泳图谱后,发现AD组大鼠与假手术组大鼠的血清中共有10个差异明显的蛋白质点,选取了2个最为明显的蛋白质点进行鉴定,分别为：血浆视黄醇结合蛋白前体,补体成分4,基因2（C4b）。结论C4b是新发现的与AD相关的蛋白质,有可能是潜在的AD标志物,有助于更深刻地理解AD的发病机制。     

蛋白质组学技术分析5-aza-2dC处理白血病细胞HL-60前后的差异表达蛋白质

目的:研究甲基化转移酶抑制剂5-杂氮-2'-脱氧胞苷(5-aza-2'-deoxycitydine, 5-aza-2dC)对人急性髓系白血病细胞HL-60细胞蛋白质表达谱的影响,探讨5-aza-2dC抗急性髓系白血病作用的机制.方法:采用二维凝胶电泳(2-DE)技术分离5-aza-2-dC处理与未处理HL-60细胞的总蛋白质,PDquest 图像分析软件识别差异蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定差异蛋白质点. 然后采用Western 印迹检测差异蛋白质半乳糖凝集素1(Galectin-1) 在药物处理与未处理HL-60细胞中的表达水平.结果: 建立了5-aza-2dC 处理与未处理HL-60 细胞蛋白质的2-DE 图谱,识别了35个差异表达的蛋白质点,鉴定了27个差异表达的蛋白质,其中包括Galectin-1在内的21个蛋白质在5-aza-2dC 处理后的HL-60 细胞中表达上调,6个蛋白质表达下调或缺失.Western 印迹结果证实Galectin-1在5-aza-2dC 处理HL-60细胞中表达上调.结论:21个表达上调蛋白质的编码基因可能是HL-60细胞的甲基化失活基因,它们可能与急性髓系白血病发病以及5-aza-2-dC抗人急性髓系白血病细胞作用有关.     

慢性间歇性缺氧大鼠肾脏组织蛋白质组的变化

目的:比较慢性间歇性缺氧大鼠与正常大鼠肾脏组织蛋白质双向电泳图谱,寻找和鉴定慢性间歇性缺氧大鼠肾脏组织中的差异蛋白表达谱。方法:以固相pH梯度等电聚焦为第一向和垂直SDS-PAGE为第二向,分别对正常对照大鼠和慢性间歇性缺氧大鼠的肾脏组织蛋白质样品进行二维分离,2-DE图谱经ImageMaster 2D Platinum V5.0软件分析,选取4个差异蛋白点用基质辅助激光解吸附离子化飞行时间质谱(MALDI-TOF-MS)进行鉴定。结果:通过对2-DE图谱蛋白斑点的匹配及对比分析,与慢性间歇性缺氧相关的差异表达蛋白斑点为112个;经质谱鉴定的4个差异表达的蛋白斑点,慢性间歇性缺氧组量下调的差异点1个,即线粒体ATP合成酶δ亚基;上调的差异点3个,分别为己糖激酶、儿茶酚-O-甲基转移酶、脱嘌呤/脱嘧啶核酸内切酶/氧化还原因子-1。结论:大鼠肾脏组织慢性间歇性缺氧损伤相关的差异蛋白表达变化可能通过多种途经影响肾脏功能。     

苯比芘作用于16HBE气道上皮细胞的蛋白质组学研究

目的应用蛋白质组学方法筛选出苯比芘（BaP）作用过的16HBE气道上皮细胞（处理组）与正常16HBE气道上皮细胞（对照组）的差异表达蛋白质,为阐明BaP对人体气道的致癌机制提供理论依据。方法培养人气道上皮细胞16HBE,并用BaP进行处理,采用双向凝胶电泳（2-DE）分离细胞总蛋白,运用 Image Master 2D Elite 5.0图象分析软件识别差异表达的蛋白质点,应用基质辅助电离解析飞行时间质谱（MALDI-TOF-MS）鉴定差异表达的蛋白质点,明确其生物学功能。结果建立双向电泳图谱,找出15个表达量有明显差异的蛋白质点,其中包括转酮醇酶、埃兹蛋白及泛素-蛋白酶体等与肿瘤发病密切相关的蛋白质。结论建立重复性较好16HBE人气道上皮细胞双向电泳图谱,并鉴定出一些与肿瘤发病机制相关的蛋白质,为阐明BaP对人体气道的致癌机制提供理论依据     

慢性肾小球肾炎肾阴虚证血浆蛋白质双向凝胶电泳分析

目的初步观察慢性肾小球肾炎肾阴虚证患者血浆蛋白质组的变化与表达差异,为进一步探讨肾阴虚证的发生机制奠定基础。方法采用双向凝胶电泳（2-DE）技术分离慢性肾小球肾炎肾阴虚证患者、肾阳虚证患者和正常人的血浆总蛋白,银染显色;PDQest软件对凝胶图像进行定性定量差异表达分析;从中选取差异表达蛋白质斑点,通过基质辅助激光解析电离飞行时间质谱（MALDI-TOF-MS）分析,获取肽质量指纹图谱（PMF）;通过Mscot-Wizard软件进行数据库搜索,鉴定蛋白质。结果与正常人和慢性肾小球肾炎肾阳虚证患者相比,在慢性肾小球肾炎肾阴虚证患者血浆蛋白质双向凝胶电泳图谱中绝对高表达的蛋白质斑点有80个,绝对低表达的蛋白质斑点有42个;定性差异表达分析发现,在慢性肾小球肾炎肾阴虚证患者血浆蛋白质双向凝胶电泳图谱中特异出现的蛋白质斑点有2个,缺失的蛋白质斑点有13个。结论同病异证患者血浆中存在差异表达蛋白质,利用蛋白质组学技术有望对中医证候的发生机制进行阐释。     

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.     

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.     

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.