氧化低密度脂蛋白及抗氧化剂对人脐静脉内皮细胞表达CD40和CD40 L的影响

目的探讨OX -LDL和VitE对人脐静脉内皮细胞表达CD40 和CD40L的影响。方法应用流式细胞技术检测细胞表面CD40和CD40L表达水平。结果低浓度OX -LDL(<200μg/L)可使内皮细胞表达CD40 和CD40L含量明显增加 ,具有浓度和时间依赖效应。高浓度(>200μg/L)OX -LDL刺激时 ,CD40 和CD40L表达明显下降。抗氧化剂VitE呈剂量依赖性部分逆转OX -LDL对内皮细胞表达CD40 和CD40L。结论OX -LDL刺激内皮细胞高表达CD40和CD40L可能是其致动脉粥样硬化的重要机制。     

人脐静脉内皮细胞表达CD137分子及其功能的初步研究

目的　了解人脐静脉内皮细胞 (HUVEC)CD137分子的表达情况 ,探讨其对内皮细胞功能的影响。方法　胰蛋白酶法分离培养人脐静脉内皮细胞 ,用RT PCR、定量荧光PCR和流式细胞仪技术 ,分别观测未刺激和用内毒素 (LPS)刺激的内皮细胞CD137mRNA与蛋白的表达情况。用anti CD137单克隆抗体交联内皮细胞表面CD137,ELISA检测其上清液中IL 8的含量。结果　①RT PCR和实时荧光定量RT PCR结果显示 ,未经刺激的人脐静脉内皮细胞表达少量CD137mRNA ,流式细胞仪检测证实其细胞表面表达少量CD137蛋白 ;当用LPS刺激内皮细胞培养 2 4h后 ,mRNA和蛋白表达量均显著提高。②ELISA检测发现 ,anti CD137单克隆抗体交联内皮细胞表面CD137分子后 ,和同型抗体对照组相比 ,其培养上清液中细胞因子IL 8的表达量明显提高 ,与LPS刺激内皮细胞上调IL 8表达的程度相当。结论　人脐静脉内皮细胞组成性表达少量CD137分子 ,LPS可以显著上调其表达。交联内皮细胞表面CD137分子可提高其IL 8产生和分泌 ,提示其具有刺激内皮细胞活化的生物功能     

B7-H3在人肝癌细胞株HepG2对外周血CD8+T细胞活化、周期及分泌IL-17调节中的作用

目的 研究B7-H3在人肝癌细胞株HepG2对人外周血CD8+T细胞活化、周期及IL-17分泌等调节中的作用.方法 RT-PCR及FCM检测B7-H3在HepG2细胞上的表达;应用脂质体法将PGPU6/GFP/neo-B7-H3shRNA质粒转入肝癌细胞株HepG2,阻断B7-H3的表达;免疫磁珠分选健康人外周血CD8+T细胞;FCM分析B7-H3分子在HepG2细胞对PHA刺激下CD8+T细胞活化、周期及PMA刺激下CD8+T细胞分泌IL-17调节中的作用.结果 肝癌细胞株HepG2高表达B7-H3分子,PGPU6/GFP/neo-B7-H3 shRNA质粒能有效阻断B7-H3在HepG2细胞上的表达;FCM分析结果显示,肝癌细胞株HepG2对CD8+T细胞活化及周期均有抑制作用;阻断B7-H3的表达后,明显减弱HepG2细胞对CD8+T细胞早期活化表型CD69表达的抑制作用,且能够通过下调CD8+T细胞Go/G1期细胞数量,上调S期细胞数量逆转HepG2细胞对CD8+T细胞周期的阻滞作用;在HepG2存在条件下,CD8+T细胞对IL-17的分泌明显增加,阻断B7-H3的表达后,IL-17的分泌被进一步上调.结论 HepG2细胞高表达B7-H3分子;B7-H3能够协同HepG2细胞对CD8+T细胞活化表型CD69的表达及细胞周期的抑制作用;HepG2细胞上调CD8+T细胞对IL-17的分泌作用,但B7-H3可抑制该上调作用.     

Bryostatin-1调节TNF-α激活的树突状细胞免疫功能

目的探讨Bryostatin-1对树突状细胞(DC)免疫功能的调节作用。方法联合应用GM-CSF和IL-4自人外周血单核细胞定向分化DC;以Bryostatin-1刺激TNF-α诱导成熟的DC,收集DC及其上清夜,以FACS和ELISA方法分析DC表面免疫分子(CD80、CD83、CD86、HLA-DR和B7-H1)和细胞因子(IFN-γ、IL-1β、IL-10和IL-12)表达水平;自DC提取RNA,以Northern blot分析DC的B7-h1 mRNA表达水平;以DC为诱导细胞,进一步检测DC的免疫诱导功能。结果Bryostatin-1通过降低DC表面B7-H1表达和增强B7.2表达,以及调节DC细胞因子(IL-1b、IL-12和IFN-γ)分泌,增强DC的免疫诱导功能,PKC通道特异性的抑制剂BI明显逆转Bryostatin-1的上述作用。结论Bryostatin-1增强DC免疫功能,其机制可能是激活PKC通道。     

小胶质细胞表达B7-H1及LPS刺激对其表达影响的研究

目的 了解共刺激分子B7-H1在小鼠中枢神经系统小胶质细胞上的表达,及LPS刺激后的表达变化情况,探讨小胶质细胞在大脑炎症状态下的免疫功能变化。方法利用小鼠小胶质细胞株N9,以LPS刺激其活化,应用RT-PCR、定量PCR、流式细胞术、免疫组化检测B7-H1在刺激前后的表达变化规律;分离培养小鼠原代小胶质细胞并进行GSA-IB4染色鉴定。应用免疫组化检测原代小胶质细胞在LPS刺激前后的表达变化。结果①小鼠小胶质细胞株N9在未刺激状态下表达B7-H1的mRNA和蛋白分子;LPS刺激细胞活化后释放肿瘤坏死因子α,一氧化氮增加,同时B7-H1的mRNA和蛋白表达水平增加。②分离培养的小鼠原代小胶质细胞经GSA-IB4染色鉴定纯度在95％以上,免疫组化显示原代小胶质细胞组成性表达B7-H1,LPS刺激后表达明显上调。结论小鼠小胶质细胞株N9小胶质细胞组成性表达共刺激分子B7-H1,LPS刺激后表达上调,提示其参与了中枢神经系统免疫应答的调节。     

人膜型LIGHT分子对Mo-DC成熟和T细胞激活的调节作用

利用我们建立的表达人膜型LIGHT分子的基因转染细胞(L929/LIGHT)探讨LIGHT/HVEM信号体外对Mo-DC诱导分化的影响,并进一步研究其对T细胞活化和抗凋亡的共刺激作用。从健康人外周血中分离的单核细胞经GM-CSF联合IL-4诱导形成Mo-DC,流式细胞术分析Mo-DC诱导过程中HVEM和LIGHT的表达;基因转染细胞L929/mock、L929/LIGHT、L929/CD40L或L929/LIGHT联合L929/CD40L,分别经丝裂霉素处理后,与GM-CSF联合IL-4诱导的Mo-DC共育,流式细胞术检测Mo-DC细胞表面成熟标志CD83和CD86的表达,利用FITC-Dextran分析Mo-DC对抗原的摄取能力;L929/LIGHT或L929/mock经丝裂霉素处理后,与抗人CD3单抗激发的T细胞共育,流式细胞术分析CD4+和CD8+T细胞表面活化标志CD25的表达,Annexin V和PI双标记分析T细胞的凋亡率。结果表明,高表达HVEM的单核细胞在诱导形成成熟Mo-D(iDC)的过程中下调了HVEM的表达,成熟Mo-DC(mDC)又上调表达HVEM,而LIGHT在Mo-DC分化过程中呈短暂的诱导性表达;基因转染细胞L929/LIGHT及其联合L929/CD40L能上调Mo-DC表面共刺激分子CD83和CD86的表达,并下调Mo-DC对FITC-Dextran的摄取能力;L929/LIGHT细胞能上调CD4+、CD8+T细胞CD25的表达,并增强T细胞抗凋亡能力。因此,基因转染细胞L929/LIGHT表面表达的人膜型LIGHT分子介导的LIGHT/HVEM共刺激信号对Mo-DC的诱导成熟和T细胞活化及抗凋亡能力具有促进作用。     

肾小管上皮细胞表达共刺激分子B7-DC并抑制CD4+T细胞活化

目的 探讨肾小管上皮细胞(TECs)表达B7-DC及其对T细胞活化的调节作用.方法 免疫组化检测肾穿刺标本B7-DC表达;流式细胞术分析人及小鼠肾小管上皮细胞B7-DC的表达变化;使用TECs/CD4 T共培养分析TECs表达的B7-DC对CD4 T细胞活化的影响.结果 慢性肾小球肾炎、狼疮性肾炎、小管间质性.肾炎等肾活检组织中发现B7-DC显著表达于肾小管.IFN-γ、TNF-a等炎症因子可诱导体外.肾小管上皮细胞表达137-DC.共培养试验发现阻断B7-DC信号可增强CD4 T细胞分泌细胞因子IFN-γ及IL-2并促进CD4 T细胞表达CD69.结论 肾小管上皮细胞表达B7-DC并可显著下调CD4 T细胞活化,在多种慢性肾脏疾病发展中可能发挥重要作用.     

LPS诱导肺微血管内皮细胞ICAM-1的表达及 NF-κ B作用的实验研究

目的研究 LPS诱导的肺微血管内皮细胞( PWVEC) ICAM- 1的表达及调控机制.方法 100ng/ml LPS刺激 PMVEC 0h、2h、4h、6h、8h或 10ng/ml、50ng/ml、100ng/ml LPS刺激 6h,免疫细胞化学检测 PMVEC ICAM- 1的表达.凝胶电泳迁移率变化分析检测 NF- κ B的活化.并通过加入活化阻断剂 PDTC观察对 PMVEC ICAM- 1表达的影响.结果 PMVEC ICAM- 1的表达与 LPS的刺激呈时相 - 剂量依赖方式.LPS的刺激迅速活化 NF- κ B,60min达到高峰,后逐渐下降.PDTC能显著降低 ICAM- 1的表达( P<0. 01).结论 LPS刺激诱导 NF- κ B的活化,启动 ICAM- 1的合成表达.     

CD40在人腹膜间皮细胞的表达

目的:对人腹膜间皮细胞CD40的表达及其调节因素进行初步探讨。方法:从CAPD患者的透出液中分离、培养腹膜间皮细胞,用IFN-γ、TNF-α、IL-1、LPS刺激24h,通过流式细胞仪(FACS)检测分析腹膜间皮细胞CD40、CD40L及ICAM-1的表达。结果:腹膜间皮细胞结构性表达少量的CD40;IFN-γ可显著增加腹膜间皮细胞表面CD40蛋白的表达,而TNF-α、IL-1、LPS对腹膜间皮细胞表面CD40蛋白的表达无显著影响。未见间皮细胞表达CD40L。IFN-γ、TNF-α、IL-1、LPS对间皮细胞ICAM-1表达均有显著增强作用。IFN-γ增强ICAM-1表达作用显著高于TNF-α、IL-1、LPS,间皮细胞CD40表达强度与ICAM-1呈显著正相关。结论:人腹膜间皮细胞可功能性表达CD40。     

TLR7、TLR9激动剂对人外周血B细胞作用的研究

人B淋巴细胞被TLR7、TLR9配体刺激后,可被激活进行克隆性增殖,促进其行使免疫功能。本研究使用TLR7、TLR9激动剂刺激健康人PBMC。使用流式细胞术,用荧光素标记的CD19筛选出B细胞后,CFSE法测定B细胞增殖情况,Annexin V测定B细胞凋亡情况,并且测定B细胞表面CD80、CD86、CD40、HLA-DR表达和B细胞IgG、IgM分泌情况。结果表明,TLR7、TLR9激动剂能显著刺激B细胞增殖、表面CD80、CD86、CD40和HLA-DR的表达上调以及刺激B细胞分泌IgM。不仅如此,TLR9激动剂还能降低B细胞早期凋亡并且刺激IgG分泌增加,而TLR7激动剂在上述方面作用却不显著。TLR9信号通路在刺激B细胞增殖、降低其早期凋亡、刺激其CD80上调以及IgG分泌方面显著强于TLR7。课题组需要进一步研究以探寻TLR9与TLR7在人B细胞内受体表达和信号转导通路的区别,为TLR通路缺陷相关疾病的发病机制和治疗研究提供理论依据。     

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.     

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.