抗CD4单抗诱导免疫耐受防治自身免疫性心肌炎的研究

目的探讨体内注射CD4单克隆抗体诱导大鼠对猪心肌肌凝蛋白产生免疫耐受及这种免疫耐受对自身免疫性心肌炎大鼠的治疗作用。方法分别于第0、7天给予Lewis大鼠体内注射猪心肌肌凝蛋白诱导自身免疫性心肌炎的产生。分别于第-2、-1、0、1天注射CD4单克隆抗体诱导免疫耐受。初次免疫后18d,检测免疫耐受的诱导情况及其对心肌炎大鼠的作用。结果同非治疗组鼠相比较,抗体治疗组鼠心功能明显改善;没有明显的心肌变性、坏死、炎症细胞浸润及纤维化;抗原特异性淋巴细胞增殖反应明显下降;血清抗心肌肌凝蛋白自身抗体明显降低;血清T_H1细胞因子IFN-γ、1L-2的水平显著下调;T_H2细胞因子IL-6、IL-10的水平没有改变或者被上调。结论CD4单抗能够诱导机体对猪心肌肌凝蛋白产生免疫耐受,通过免疫耐受的诱导阻止了自身免疫性心肌炎大鼠心功能的紊乱和心肌的损伤。     

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     

腺病毒载体介导的共刺激分子融合蛋白对实验性自身免疫性心肌炎的作用

目的探讨腺病毒载体介导的共刺激分子融合蛋白CTLA4Ig与ICOSIg联合治疗对实验性自身免疫性心肌炎（EAM）的作用.方法猪心肌肌球蛋白免疫Lewis大鼠制成EAM模型.分别构建CTLA-4胞外域、ICOS胞外域与人IgG Fc段融合的腺病毒表达载体,常规方法生产表达上述融合蛋白的腺病毒用于治疗,免疫第14天注射后观察至第28天.第28天超声心动图检测心脏功能,苏木素-伊红（HE）染色观察心肌炎症程度,Western blot检测心肌CTLA-4、ICOS、ICOSL及B7-1、B7-2蛋白表达水平,ELISA检测血浆IL-2、IL-4和IFN-γ水平.结果CTLA4Ig、ICOSIg单独或联合治疗均使大鼠心功能指标、心肌炎症程度明显改善.Western blot显示联合治疗组CTLA-4、ICOSL及ICOS、B7-1蛋白表达下调,而B7-2表达差异无统计学意义.细胞因子平衡向TH2方向偏离.结论CTLA4Ig及ICOSIg联合阻断共刺激分子通路减轻EAM自身免疫性心肌损伤,改善大鼠心脏功能.其机制可能通过下调心肌组织CTLA-4、ICOS、ICOSL及B7-1蛋白表达.     

非诺贝特通过调节Th1/Th2细胞因子的平衡改善自身免疫性心肌炎

目的探讨非诺贝特(fenofibrate)对自身免疫性心肌炎(EAM)大鼠的治疗作用。方法将提纯精制后的猪心室肌球蛋白加等体积含灭活的结核杆菌的完全弗氏免疫佐剂充分混匀后,于Lewis大鼠双后足皮下注射制作大鼠EAM模型。于第0天免疫后,非诺贝特组给予非诺贝特灌胃,EAM组给予生理盐水灌胃。免疫后的第17天,超声检测大鼠心功能,处死大鼠取心脏评估心肌炎的严重程度及心体质量比,HE染色,应用实时定量PCR检测心脏的IFN-γ,IL-2,IL-4和IL-10的mRNA表达水平,并进一步通过ELISA检测血清中IFN-γ和IL-4的表达水平。结果非诺贝特组大鼠的心功能指标与EAM组相比得到了显著改善。非诺贝特治疗组的心体质量比和心肌的炎症浸润程度也比EAM模型组有明显的降低。另外,非诺贝特治疗组降低了Th1的细胞因子(IFN-γ,IL-2)的表达并增加了Th2细胞因子(IL-4,IL-10)的表达。ELISA的结果也显示非诺贝特治疗组血清中IFN-γ的水平降低而IL-4的水平升高。结论非诺贝特治疗可能通过调节Th1/Th2细胞因子的平衡改善自身免疫性心肌炎。     

肌球蛋白诱导大鼠自身免疫性心肌炎不同时期心功能及心肌病理学变化

目的:观察心肌肌球蛋白诱导自身免疫性心肌炎不同时间实验大鼠心功能及心肌组织病理学动态变化。方法:以猪心肌肌球蛋白为致病性抗原,与完全弗氏佐剂等体积混合成乳浊液在实验第1天、8天、30天皮下注射免疫Lewis大鼠作为心肌炎组,以完全弗氏佐剂皮下注射大鼠作为对照组。HE染色和Masson染色观察两组大鼠初次免疫后第21天、30天、90天心肌病理变化,超声心动图检测两组大鼠初次免疫后第21天、30天、90天的射血分数(EF)、左室短轴缩短分数(FS)、左室舒张末内径(LVEDd)、左室收缩末内径(LV-EDs)及左心室后壁厚度(LVPW)等指标。结果:免疫后不同时间超声心动图检测提示心肌炎组大鼠心功能较对照组明显下降(EF:84.25±2.17%vs93.45±4.13%,P<0.05;FS:48.49±4.36%vs63.17±4.49%,P<0.05;LVEDd:3.66±0.35mmvs4.63±0.32mm,P<0.05)。免疫后第21天,HE染色见心肌炎组大鼠心肌细胞肿胀、变性、坏死及不同程度的急性炎性细胞浸润,至第90天心肌组织炎症减弱,间质出现纤维化,胶原容积分数升高,对照组大鼠未出现心肌炎症及纤维化。结论:心肌肌球蛋白可诱导大鼠实验性自身免疫性心肌炎,急性期心功能受损主要由心肌炎症引起,慢性期病理学改变主要表现为心肌纤维化。     

IL-1RⅡ重组质粒导入对大鼠实验性自身免疫性心肌炎的治疗作用

目的: 评估编码Ⅱ型白细胞介素-1受体(type Ⅱ interleukin-1 receptor,IL-1RⅡ)的重组质粒在大鼠实验性自身免疫性心肌炎 (experimental autoimmune myocarditis,EAM)中的治疗效果。方法: 将猪心室肌球蛋白与等体积完全弗氏免疫佐剂充分混匀后,于第0 d Lewis大鼠双后足皮下注射制作EAM模型,第6 d应用流体动力学方法进行重组质粒IL-1RⅡ的体内导入,第17 d进行大鼠心脏超声检查后,处死大鼠,检测大鼠心/体重比值、心脏病理学评估及心肌炎面积率,进一步应用实时荧光定量RT-PCR检测心衰标记物脑钠尿肽(brain natriuretic peptide,BNP)的水平。同时检测白细胞介素-1(interleukin-1,IL-1)相关因子白细胞介素-1β(interleukin-1 beta,IL-1β)、前列腺素E₂合成酶(prostaglandin E₂ synthase, PGES)以及单核细胞趋化蛋白-1(monocyte chemotactic protein-1, MCP-1) mRNA及蛋白表达水平。结果: IL-1RⅡ重组质粒导入后次日表达量最高,且升高持续实验整个过程至第17 d。 IL-1RⅡ治疗组的大鼠心/体重比值、心肌炎面积率、BNP及IL-1表达均较对照组有明显下降,大鼠左室心功能亦有明显改善。结论: 应用流体动力学方法进行重组质粒IL-1RⅡ的体内导入治疗,对大鼠实验性自身免疫性心肌炎有改善左室功能和阻止心肌损害作用。     

静脉注射IL-19重组质粒对大鼠实验性自身免疫性心肌炎的治疗作用

目的:评估静脉注射白细胞介素19(interleukin-19,IL-19)重组质粒对大鼠实验性自身免疫性心肌炎(experimental autoimmune myocarditis,EAM)的治疗作用。方法:将猪心室肌球蛋白与等体积完全弗氏免疫佐剂充分混匀后,双足皮下注射制作大鼠EAM模型,免疫后第6天应用静脉注射方法将IL-19重组质粒导入体内,第17天行心脏超声检查后处死大鼠,检测心脏重量与体重比值、病理学评估、心肌炎面积率;实时荧光定量PCR检测心衰标记物心房钠尿肽(atrial natriuretic peptide,ANP)、脑钠尿肽(brain natriuretic peptide,BNP)的表达水平,进一步检测心肌相关炎症因子IL-18、IL-1β、IL-12p35和IFN-γ的表达水平。结果:IL-19重组质粒导入组的大鼠心功能得到显著改善;心脏重量体重比、心肌炎面积率、ANP、BNP的表达均较模型组明显下降;相关炎症细胞因子的表达也显著降低。结论:应用静脉注射法进行IL-19重组质粒体内导入治疗,明显抑制大鼠实验性自身免疫性心肌炎的炎症反应,减轻了心肌损伤从而改善心功能。     

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。     

TNF-α和cTnI在BALB/c小鼠自身免疫性心肌炎模型的动态变化

目的：以包含心肌球蛋白致病性抗原表位的人工合成多肽免疫BALB／c小鼠，建立实验性自身免疫性心肌炎动物模型，观察其自身免疫阶段和慢性病变期的病理变化，检测炎性因子和自身抗体变化。方法：人工合成含心肌肌球蛋白抗原重链表位长度为30个氨基酸残基的多肽0．8mg，纯化多肽皮下免疫遗传易感BALB／c小鼠，建立自身免疫性心肌炎模型，检测TNF—α和cTnl病变期间的改变。结果：实验组小鼠出现自身免疫性心肌炎病理改变。第14d出现心肌损伤，至第21d～30d心肌损伤达高峰，60d趋向稳定。心脏扩大，病理切片可见心肌层充血，间质巨噬细胞浸润明显，心肌细胞肿胀、逐步崩解坏死，被纤维组织取代。血清出现抗心肌肌球蛋白自身抗体阳性，并伴TNF—α和cTnl水平升高、高峰、下降的变化。结论：可成功诱导BALB／c系小鼠发病，建立实验性自身免疫性心肌炎动物模型，TNF—α和cTnl水平反映自身免疫性心肌炎病情发生、发展和转归。     

TNF-α和cTnI在BALB/c小鼠自身免疫性心肌炎模型的动态变化

目的:以包含心肌球蛋白致病性抗原表位的人工合成多肽免疫BALB/c小鼠,建立实验性自身免疫性心肌炎动物模型,观察其自身免疫阶段和慢性病变期的病理变化,检测炎性因子和自身抗体变化.方法:人工合成含心肌肌球蛋白抗原重链表位长度为30个氨基酸残基的多肽0.8mg,纯化多肽皮下免疫遗传易感BALB/c小鼠,建立自身免疫性心肌炎模型,检测TNF-α和cTnI病变期间的改变.结果:实验组小鼠出现自身免疫性心肌炎病理改变.第14d出现心肌损伤,至第21d～30d心肌损伤达高峰,60d趋向稳定.心脏扩大,病理切片可见心肌层充血,间质巨噬细胞浸润明显,心肌细胞肿胀、逐步崩解坏死,被纤维组织取代.血清出现抗心肌肌球蛋白自身抗体阳性,并伴TNF-α和cTnI水平升高、高峰、下降的变化.结论:可成功诱导BALB/c系小鼠发病,建立实验性自身免疫性心肌炎动物模型,TNF-α和cTnI水平反映自身免疫性心肌炎病情发生、发展和转归.     

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.