应用基因芯片研究肾移植急性排斥发生时外周血免疫基因变化

目的：借助基因芯片研究肾移植受者外周血淋巴细胞免疫相关基因在急性排斥反应中的作用。方法：8例肾移植术后发生急性排斥反应患者，于手术和肾穿刺当日收集外周血作为对照血样和实验血样。用Ficoll技术分离外周血淋巴细胞，并采用一步法提取总RNA，逆转录合成cDNA探针，荧光染料标记后，采用免疫相关基因芯片进行芯片杂交，扫描后筛选出差异表达基因。结果：急性排斥发生时外周血淋巴细胞免疫相关基因差异表达49条，其中上调25条，下调24条。结论：从基因水平上，外周血淋巴细胞在肾移植急性排斥的免疫应答中涉及不同环节，免疫抑制剂在这些环节上对基因的表达存在不同程度的影响。     

应用基因芯片研究肾移植急性排斥中外周血淋巴细胞免疫相关基因变化

目的：借助基因芯片研究肾移植受者外周血淋巴细胞在急性排斥的免疫应答中的作用。 方法： 8例肾移植术后发生急性排斥反应患者，于手术和肾穿刺当日收集外周血作为对照血样和实验血样。用Ficoll技术提取外周血淋巴细胞，并采用一步法提取总RNA，逆转录合成cDNA探针, 荧光染料标记后，采用免疫相关基因芯片进行芯片杂交，扫描后筛选出差异表达基因。 结果： 急性排斥发生时外周血淋巴细胞免疫相关基因差异表达49条，其中上调25条，下调24条。 结论： 从基因水平上，外周血淋巴细胞在肾移植急性排斥的免疫应答中涉及不同环节，免疫抑制剂在这些环节上对基因的表达存在不同程度的影响。     

供体骨髓输注减轻同种肝移植排斥反应

目的:研究非清髓性骨髓移植在同种肝移植免疫反应中的作用及其意义。 方法:动物实验为同种大鼠肝移植：Ⅰ组为Wistar→Wistar同基因对照组；Ⅱ组为SD→Wistar急性排斥组；Ⅲ组为SD→Wistar环孢素A（cyclosporine A,CsA）肌肉注射处理组；Ⅳ组为SD→Wistar术前1周大剂量供体骨髓细胞输注处理组。用术后一般状态、生存时间、病理学排斥分级、肝标本中IL-2、IFN-γ的表达以及胸腺内嵌合体的检测等手段，确定不同处理组移植后的排斥反应情况及机体的免疫状态。 结果:Ⅰ组大鼠术后无排斥反应发生；Ⅱ组大鼠术后9-13 d内全部死亡，中位存活时间（10.7±0.5）d，排斥反应明显；Ⅲ组大鼠仅见轻度排斥；Ⅳ组自然存活亚组6只大鼠4只长期存活 (>100 d)，排斥分级明显低于Ⅱ组（P<0.05）,与Ⅱ组相比，Ⅳ组IL-2、IFN-γ蛋白的表达均明显下调。在骨髓输注15 d后，用PCR法在雌性Wistar大鼠受体胸腺内可检测到雄性SD大鼠供体特异性的Y染色体（Y-chromosome-specific sequence，Sry）基因片段。 结论:供体骨髓输注可形成供受体细胞的嵌合状态，可降低同种异体肝移植的排斥反应，延长受体生存时间。     

大鼠ARDS基因表达谱中信号传导基因的表达

 目的 对大鼠急性呼吸窘迫综合征(ARDS)基因表达谱中的信号传导基因进行研究与分析。方法 从正常和ARDS大鼠肺组织中提取总RNA,分离纯化mRNA,经反转录合成掺入生物素标记的cDNA探针,然后与基因芯片杂交,扫描芯片荧光信号图像,用Sam3.0进行统计处理,用博奥生物分子功能注释系统V4.0进行功能分析。随机选择3个差异表达基因 ,用荧光定量RT- PCR验证。结果 在ARDS大鼠肺组织中,信号传导相关基因上调的有2个,下调的有9个,涉及到的相关信号通路11个。结论 大鼠ARDS基因表达谱中涉及到许多信号传导基因和通路的差异表达。     

大鼠肝移植急性排斥反应模型的建立

背景：研究表明Wistar和SD大鼠普遍为封闭杂交系而非近交系,有一定的遗传稳定性,同时也有一定的基因多态性,因此Wistar和SD大鼠之间的肝移植模型可能不是研究大鼠急性排斥反应的理想模型。 目的：建立Lewis-BN大鼠肝脏移植急性排斥反应模型。 方法：采用改良的“Kamada”二袖套法,分别进行同基因Lewis-Lewis间肝移植及Lewis-BN间肝移植。移植后3,5,7 d观察受体肝脏组织病理变化,测定谷丙转氨酶和总胆红素水平变化及大鼠存活时间。 结果与结论：同基因移植组大鼠无急性排斥表现,平均存活时间超过100 d,肝功能损害较轻。异基因移植组大鼠存活 (12.75±1.25) d,移植后第7 天肝脏病理检查有明显的排斥反应,肝功能损害较重,移植后各时相点谷丙转氨酶和总胆红素水平均明显高于同基因移植组(P < 0.05)。提示Lewis-BN大鼠肝移植组合为稳定的大鼠肝移植急性排斥模型,但近交系大鼠对手术耐受性差,建模难度大,熟练的显微外科技术和细心轻柔的操作是模型成功的关键。     

调节性T细胞在大鼠小肠移植急性排斥反应中的作用

目的 分析调节性T细胞在大鼠急性排斥反应中的作用。方法 采用免疫组化SABC染色法，测定BN→LEW大鼠小肠移植急性排斥反应时，外周血及移植肠浸润淋巴细胞中调节性T细胞：CD4^ ,CD8^ ,CD25^ T淋巴细胞及相关细胞因子IL－4和IFN－γ的表达，并与同基因大鼠间小肠移植（BN→BN）作比较。结果 外周血淋巴细胞分析显示，大鼠小肠移植急性排斥反应时，以CD4^ ,CD25^ ,T细胞为主，CD8^ 淋巴细胞只占少部分；分泌IL－4的细胞在术后4，7，14d分别只占14．3％，16．2％和16．9％。移植肠基底浸润的淋巴细胞以CD4^ ,CD25^＋和分泌IFN－γ的淋巴细胞为主。结论 在大鼠同种小肠移植中，急性排斥反应与CD25^ ,CD4^ T细胞及Th1相关细胞因子IFN－γ的表达增加相关。而CD8^＋T淋巴细胞和Th2相关细胞因子IL－4可能具有保护作用。     

应用基因芯片筛选抑郁症相关基因的初步研究

目的 筛选与抑郁症相关的差异表达基因.方法 采用人8000条cDNA表达谱基因芯片技术,研究抑郁症患者及健康对照组外周血淋巴细胞基因表达差异,从而筛选抑郁症相关基因.结果 与对照组相比,抑郁症组中存在显著性表达差异的基因有138个,其中上调有78个,下调有60个基因.结论 离子通道、免疫活性、蛋白代谢、能量代谢(糖脂代谢)、细胞增殖、细胞黏附以及信号传导等相关基因类型可能与抑郁症密切相关.     

供体特异性输血诱导CD8+CD28-调节性T细胞在肝移植急性排斥反应中的作用

目的 通过供体特异性输血(DST)诱导机体产生抗原特异性CD8+CD28-调节性T细胞(CD8+CD28-Tr),评价其对大鼠肝移植急性排斥反应的抑制作用.方法 采用DST在正常BrownNorway(BN)大鼠体内诱导针对LEW抗原的CD8+CD28-Tr,体外验证其免疫抑制活性之后将其输注给LEW→BN肝移植急性排斥模型受体,观察输注后大鼠的生存时间和肝脏病理学表现.采用SPSS11.0软件进行Log-Rank检验,比较大鼠生存率的差异.结果 DST可在正常BN大鼠体内诱导大量CD8+CD28-Tr产生,其在体外具有免疫抑制活性,体内输注可缓解大鼠肝移植急性排斥反应(LEw→BN).结论 DST可以在正常大鼠体内诱导大量CD8+CD28-Tr产生,其体内输注可以抑制大鼠肝移植急性排斥反应,DST可能有望成为体内诱导特异性CD8+CD28-Tr的途径之一.     

胸腺接种肝特异性抗原减轻移植肝细胞凋亡的发生

目的：研究肝特异性抗原（LSA）胸腺接种对移植肝细胞凋亡的影响。 方法： 实验为同种肝移植：Ⅰ组为Wistar→Wistar同基因对照组；Ⅱ组为SD→Wistar急性排斥组；Ⅲ组为SD→Wistar术前1周供体肝特异性蛋白受体胸腺内注射处理组。观察手术后一般状态、生存时间、肝细胞凋亡及肝组织T细胞活化连接蛋白（LAT）的表达情况，确定移植后的受体的生存情况及机体的免疫状态。 结果： Ⅰ组大鼠术后一般状态好，均长期存活；Ⅱ组大鼠体重进行性减轻，术后9-13 d内全部死亡；Ⅲ组大鼠术后一般状态同Ⅰ组，明显好于Ⅱ组，存活率明显高于Ⅱ组；实验组各时段凋亡肝细胞数均与同时段对照组无显著差异，明显低于SD-Wistar急性排斥组；实验组与对照组移植肝中均未见LAT的表达。 结论： 移植前胸腺接种供体LSA对移植肝细胞具有明显的保护作用。     

Lovastatin对肝癌HepG2细胞作用后基因差异表达的初步研究

目的： 用cDNA芯片观察lovastatin作用前后HepG2细胞的差异表达基因。 方法： 20 μmol/L的lovastatin作用于HepG2细胞18 h，各提取实验组与对照组细胞总RNA，用cDNA直接标记法制备探针，进行杂交，杂交信号经扫描后，用软件分析基因表达情况。应用RT-PCR验证部分差异表达基因。 结果： Lovastatin作用后表达上调的基因有30个，表达下调的基因有11个，涉及信号转导、肿瘤免疫、细胞周期等方面，RT-PCR结果符合基因芯片结果。 结论： 用基因芯片筛选出的lovastatin作用后的肝癌细胞差异表达基因为深入研究他汀类药物的抗肿瘤机制提供重要的理论依据。     

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.     

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 case for allele‐specific recognition by the TCR

There is a sharp difference in how one views TCR structure–function–behaviour dependent on whether its recognition of major histocompatibility complex‐encoded restriction elements (R) is germline selected or somatically generated. The generally accepted or Standard model is built on the assumption that recognition of R is by the V regions of the αβ TCR, which is not driven by allele specificity, whereas the competing model posits that recognition of R is allele‐specific. The establishing of allele‐specific recognition of R by the TCR would rule out the Standard model and clear the road to a consideration of a competing construct, the Tritope model. Here, the case for allele‐specific recognition (germline selected) is detailed making it obvious that the Standard model is untenable.