大鼠自身免疫性心肌炎模型的建立

目的: 建立Lewis大鼠实验性自身免疫性心肌炎模型。方法: 从猪心中获得心肌肌凝蛋白,以此蛋白免疫Lewis大鼠,建立自身免疫性心肌炎模型，于初次免疫后第18 d、30 d、49 d进行心脏超声及血流动力学检查，ELISA法检测血清细胞因子，取心肌组织进行组织病理学检查。结果: 心脏超声及血流动力学检查显示，免疫组鼠心功能明显低于正常对照组，以第18 d最明显。血清学检查显示IFN-γ和IL-2水平在急性期达到高峰，IL-6、IL-10水平则在恢复期达到高峰。心脏病理检查显示，初次免疫后18 d，免疫组鼠全部出现明显的心肌炎病理改变。HE染色显示初次免疫后21 d大量炎性细胞浸润，伴有心肌的坏死，第30 d炎性细胞有所减少，第49 d炎性细胞基本消失。Massine trichrome染色显示炎症区域逐渐由纤维组织代替。结论: 猪心肌肌凝蛋白可以诱导Lewis大鼠出现典型的自身免疫性心肌炎，炎症损伤在心肌炎的发生和发展中起重要作用，参与了心功能紊乱和心室重构过程。     

Toll样受体3信号通路介导自身免疫性心肌炎炎症反应及作用机制

目的探讨自身免疫性心肌炎大鼠心肌组织Toll样受体3(TLR3)表达水平及临床意义。方法将54只大鼠按照随机数字表法分为AM组、干预组和对照组,每组18只,对照组大鼠正常喂养,AM组、干预组建立自身免疫性心肌炎大鼠模型,干预组于模型建立第21天注射0.1 mg TLR3配体溶液,AM组、对照组大鼠注射等量PBS溶液。分别于模型建立第7、14、21、24、28、35天处死各组3只大鼠,观察各组大鼠心肌组织病理变化,酶联免疫吸附试验检测血清心肌肌凝蛋白自身抗体滴度,免疫组织化学染色法检测心肌组织TLR3蛋白表达,实时定量PCR检测心肌组织TLR3、TNF-αm RNA表达。结果对照组大鼠血清心肌肌凝蛋白自身抗体在不同时间点比较差异无统计学意义(P0.05),干预组血清心肌肌凝蛋白自身抗体滴度在第28天达高峰,AM血清心肌肌凝蛋白自身抗体滴度在第21天达高峰;第14、21、24、28、35天AM组、干预组血清心肌肌凝蛋白自身抗体滴度显著高于对照组(P0.05),第24、28、35天干预组血清心肌肌凝蛋白自身抗体滴度显著高于AM组(P0.05)。对照组TLR3蛋白表达在不同时间点比较差异无统计学意义(P0.05);第14、21、24、28、35天,AM组、干预组TLR3蛋白表达显著高于对照组(P0.05),第24、28、35天,干预组TLR3蛋白表达显著高于AM组(P0.05)。对照组TLR3、TNF-αm RNA相对表达量在不同时间点比较差异无统计学意义(P0.05);第7、14、21、24、28、35天,AM组、干预组TLR3、TNF-αm RNA相对表达量显著高于对照组(P0.05),第24、28、35天,干预组TLR3、TNF-αm RNA相对表达量显著高于AM组(P0.05)。结论自身免疫性心肌炎心肌组织TLR3表达上调,TLR3信号通路介导的炎症反应参与了自身免疫性心肌炎的发病。     

实验性自身免疫性心肌炎小鼠模型研制

目的 :用BALB/c小鼠建立自身免疫性心肌炎动物模型。方法 :从新鲜猪心中分离提纯心肌肌凝蛋白。在第 1,第 8和第 3 0天用心肌肌凝蛋白和完全弗氏佐剂混合的乳浊液皮下注射免疫实验组小鼠 ,而在对照组仅用完全弗氏佐剂皮下注射。于初次免疫后的第14、第 2 1、第 3 0和第 60天收集血液和心脏标本 ,进行肌凝蛋白抗体和组织病理学检查。结果 :模型组第 14天到第 3 0天的标本中有不同程度的炎性反应和心肌细胞变性坏死 ,而 60天时出现了纤维化。同时在模型组小鼠血清中检测到肌凝蛋白抗体。结论 :通过免疫心肌肌凝蛋白发生自身免疫性心肌炎的BALB/c小鼠可作为良好的研究心肌炎和扩张型心肌病发病机制的动物模型     

HVEM过表达树突状细胞对自身免疫性心肌炎的作用

目的 探讨单纯疱疹病毒进入介导子(herpes virus entry mediator,HVEM)基因修饰的树突状细胞(dendritic cells,DC)能否防治肌凝蛋白诱导的小鼠心肌炎模型,并分析其机制.方法 以肌凝蛋白加弗氏佐剂免疫小鼠制备实验性自身免疫性心肌炎(EAM)动物模型,通过小鼠尾静脉输注肌凝蛋白冲击的HVEM基因修饰的DC,观察其对心肌炎的免疫保护作用,并探讨其可能机制.结果 HVEM基因修饰的DC能够抑制自身免疫应答造成的小鼠心肌损伤,其机制主要是通过分泌IL-10,并诱导产IL-10调节性T细胞(Tr1)抑制自身反应性T细胞的活化.结论 HVEM基因修饰的DC能抑制EAM的发生,并且HVEM介导的信号网络可能在不同的细胞中产生不同的作用.     

IL-2协同抗CD45RB抗体调节Treg/Th17细胞的比例并延长小鼠移植皮肤存活时间

目的:研究IL-2在抗CD45RB抗体诱导免疫耐受中对Treg/Th17细胞分化的影响,进一步阐明抗CD45RB抗体诱导免疫耐受的机制。方法:用免疫磁珠分选C57BL/6小鼠脾脏中的CD4+T细胞,在抗CD45RB抗体与IL-2的作用下培养72小时后,流式检测Treg/Th17细胞的变化。以BALB/c小鼠为供体,C57BL/6小鼠为受体建立同种异基因皮肤移植模型,分别给予抗CD45RB抗体及IL-2等治疗,术后1、3、5、7、9天取受体鼠脾细胞,动态检测Treg/Th17细胞的变化;术后第9天取移植皮肤HE染色观察炎性细胞的浸润情况;观察并记录移植皮肤的存活时间。结果:CD4+T细胞在IL-2联合抗CD45RB抗体的作用下培养72小时后,Treg比例升高,Th17细胞比例下降;IL-2联合抗CD45RB抗体治疗后明显延长小鼠移植皮肤的存活时间。结论:IL-2可以明显增强抗CD45RB抗体诱导免疫耐受的形成,使Treg细胞上调,下调Th17细胞,有利于免疫耐受的形成。     

调节性B细胞与免疫耐受

B细胞的典型特征是可产生包括自身抗体的各种抗体.然而,B细胞还有其他的免疫功能,例如产生各种细胞因子和次级抗原递呈[1].与T细胞一样,B细胞中也有致病性和调节性的细胞亚群.研究发现抗CD45RB抗体可诱导稳定的免疫耐受模型,在诱导免疫耐受时需有宿主B淋巴细胞的参与[2].最近研究表明,体内确实存在能产生白细胞介素10(IL-10)或转化生长因子β1(TGF-β1)等细胞因子的调节性B细胞(regulatory B cells,Bregs).Bregs通过产生抑制性细胞因子或分泌抑制性抗体影响其他免疫细胞,在免疫耐受中发挥作用[1].本文将对Bregs在免疫耐受中的作用机制做一综述.     

体外DCs在抗CD45RB抗体诱导免疫耐受中的作用机制

目的:探讨树突状细胞(dendritic cells,DCs)在抗CD45RB抗体诱导的免疫耐受中所发挥的作用,从而阐明抗CD45RB抗体诱导免疫耐受的机制。方法:采用DCs的常规诱导方法(rmGM-CSF、IL-4和LPS),在诱导过程中加入不同剂量的抗CD45RB抗体,成熟后利用流式细胞仪检测细胞表型、周期和吞噬能力,ELISA法检测IL-12分泌量,混合淋巴细胞培养检测DCs对T细胞增殖能力的影响。结果:DCs经抗CD45RB抗体处理后,CD11C和CD83表达升高,CD86表达下降,自身增殖和吞噬能力增强,但分泌IL-12和刺激T细胞增殖的能力明显下降。结论:耐受性树突状细胞(tolerogenic dendritic cells,tDCs)能显著抑制T细胞的增殖,它的产生是抗CD45RB抗体诱导免疫耐受的主要机制之一。     

腺病毒介导反义CⅡTA基因对实验性自身免疫性心肌炎的治疗

目的　研究反义CⅡTA基因对心肌肌凝蛋白诱导的实验性自身免疫性心肌炎 (experi mentalautoimmunemyocarditis,EAM)的治疗作用。方法　以携带反义CⅡTA基因的腺病毒载体对肌凝蛋白免疫后的小鼠进行静脉注射 ,同时设立空载病毒组与单纯免疫组作为对照 ,以多种手段观察反义CⅡTA基因对EAM的治疗效果。结果　治疗组与两对照组相比 ,心肌病理损伤程度减轻 ;血浆抗心肌肌凝蛋白抗体滴度及cTnⅠ浓度明显下降 (P <0 .0 5 ) ,流式细胞术检测外周血与脾脏T细胞I Ad表达量明显下降 (P <0 .0 5 )。结论　反义CⅡTA基因对心肌肌凝蛋白诱导的EAM具有较好的治疗作用。     

体外急性心肌梗死早期诊断方法的研究

从人心肌中提取心肌肌凝蛋白轻链(CMLC),免疫Balb/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞做常规融合,制备单克隆抗体.通过抗人心肌肌凝蛋白轻链单克隆抗体的相加指数、亲和常数测定和抗体夹心实验,筛选到配对较好的两对单抗,在此基础上建立起轻链夹心ELISA方法,为临床急性心肌梗死早期诊断方法研究提供条件.     

人IL-2信号肽和大鼠心肌肌凝蛋白α重链片段融合基因的构建及分泌表达

目的：克隆编码大鼠心肌肌凝蛋白α重链aa736-960的cDNA片段，构建携带融合基因的逆转录病毒分泌表达载体。方法：以RT-PCR方法从幼龄大鼠心肌组织中扩增出681bp的目的基因，与hIL-2cDNA序列拼接，构建融合基因Hil-2/myosin及其逆转录病毒载体。采用脂质体法转染NIH3T3和PA317细胞，经G418筛选后，用RT-PCR、免疫组化、免疫电镜和Dot-blot法分析检测融合基因的表达。结果：核苷酸序列测定显示克隆基因的核苷酸及推导的氨基酸序列与报告序列一致，开放阅读框正确；RT-PCR检测提示转染细胞有融合基因mRNA转录；免疫组化、免疫电镜和Dot-blot检测表明转染细胞有该肌凝蛋白重链片段的分泌表达。结论：大鼠心肌肌凝蛋白α重链基因逆转录病毒载体构建成功，并可分泌表达，为心肌肌凝蛋白转基因诱导机体特异性免疫耐受及对心肌免疫损伤的研究打下了基础。     

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.