SDF-1/CXCR4在滋养层细胞中的表达及其在母-胎免疫耐受中的作用

目的:研究趋化因子SDF1及其配体CXCR4在滋养层细胞中的表达及其在母胎免疫耐受中的作用。方法:取早孕期的绒毛,分离纯化培养绒毛外滋层养细胞(extravilloustrophoblast,EVT),用免疫细胞化学染色法检测SDF1与CXCR4在绒毛中的表达。用流式细胞术筛选源于滋养层细胞高表达CXCR4的绒癌细胞株用于体外微孔隔离室迁移实验,以分析SDF1的趋化活性。用免疫组织化学染色法检测早孕期绒毛及足月妊娠胎盘中SDF1及CXCR4的表达。结果:在EVT中可检出SDF1和CXCR4的表达,在一定范围内,SDF1的趋化活性与其浓度呈正相关(r=0.68,P<0.01)。10μg/L的SDF1趋化作用最强,最大趋化指数CI为1.62±0.12。在早孕期的绒毛及足月胎盘中,滋养层细胞的胞膜和细胞质中均检出SDF1和CXCR4的表达,但在足月胎盘中的表达强度明显低于早孕期的绒毛组织(P<0.01)。结论:SDF1/CXCR4在妊娠中发挥着重要作用,对维系母胎免疫耐受具有重要意义。     

CXCL12 在母胎界面的表达及其促进BeWo滋养细胞系的体外迁移作用

目的研究CXCL12在早孕期母胎界面的表达情况及其对BeWo细胞的体外迁移作用.方法免疫组织化学法检测早孕期绒毛及蜕膜组织中CXCL12的表达情况;用Transwell细胞侵入系统检测CXCL12对BeWo细胞迁移的影响.结果早孕期绒毛及蜕膜组织均有CXCL12的表达,分布于绒毛柱、滋养细胞及间质血管内;CXCL12对BeWo细胞具有趋化作用,浓度达10ng/ml时趋化效应最强.结论 CXCL12可促进BeWo细胞的迁移,因此早孕期母胎界面产生的CXCL12可能参与滋养细胞的迁移,对于维系正常妊娠发挥了一定作用.     

人早孕滋养细胞条件培养液对外周血单个核细胞的趋化效应

目的:探讨人早孕滋养细胞在蜕膜免疫细胞选择性募集中的可能机制。方法:分离纯化人早孕滋养细胞进行原代培养,制备滋养细胞条件培养液(CM);将CM倍比稀释加入Transwell下室,分离的人外周血单个核细胞加入上室进行趋化实验,流式细胞术分析CM对PBMC各型免疫细胞的趋化效应。结果:4倍稀释的CM即显示对外周CD56+CD16-NK细胞、CD56+CD16+NK细胞、单核细胞的趋化活性,随CM浓度上升,各细胞的趋化活性也升高(P0.001);CM原液也可明显趋化外周T细胞和γδT细胞(P0.05);各浓度CM对NKT细胞均不表现趋化作用。CM趋化后的免疫细胞组成与趋化前显著不同,而与蜕膜免疫细胞的组成相似。结论:人早孕滋养细胞能够分泌多种趋化因子,募集外周血免疫细胞到达母胎界面,帮助形成蜕膜独特的免疫微环境,并维持母胎免疫耐受。     

环孢素A对人蜕膜免疫细胞分泌IL-10与IFN-γ的调控作用

探讨环孢素A(CsA)对人早孕期母胎界面主要组成细胞IL-10与IFN-γ分泌的调节作用,为反复自然流产等妊娠疾患的治疗提供新线索。收集6~10孕周行人工流产术的正常妊娠妇女绒毛和蜕膜组织;分离、培养蜕膜免疫细胞、滋养细胞与蜕膜基质细胞;ELISA法检测给予环孢素A(0μmol/L,1μmol/L)24、48、72 h后蜕膜免疫细胞、滋养细胞及蜕膜基质细胞培养上清中IL-10与IFNγ-分泌水平。结果显示,(1)1μmol/L CsA可以明显促进人早孕期蜕膜免疫细胞分泌IL-10(P<0.01),并抑制其产生IFN-γ(P<0.01);1μmol/L CsA不影响原代培养的人早孕期滋养细胞与蜕膜基质细胞IL-10的分泌(P>0.05);不论是否给予CsA处理,原代培养的人早孕期滋养细胞与蜕膜基质细胞均不产生IFN-γ。结果表明,CsA通过促进蜕膜免疫细胞分泌IL-10,并抑制其产生IFNγ-,从而有利于母胎界面形成Th2型免疫优势。     

母-胎界面趋化因子及其受体表达与作用

母-胎界面的滋养细胞、蜕膜淋巴细胞和蜕膜基质均表达多种趋化因子及其受体。滋养细胞分泌的MIP-1α募集外周血中CD56brightNK细胞至母-胎界面;蜕膜血管表达SLC,特异性趋化CD56brightNK细胞。除募集蜕膜淋巴细胞外,母-胎界面的趋化因子及其受体尚参与固有免疫应答,并调节滋养细胞侵袭、分化及胎盘形成。滋养细胞固有表达CXCR4和CCR5,HIV-1利用CXCR4或CCR5入侵胎儿细胞,导致HIV-1经子宫垂直传播。滋养细胞及蜕膜细胞分泌的IL-8在炎症相关的早产中具有重要作用。     

人早孕蜕膜基质细胞及免疫活性细胞趋化因子受体CXCR6的表达

目的分析人早孕期蜕膜基质细胞趋化因子配基受体对CXCL16/CXCR6的表达及免疫活性细胞趋化因子受体CXCR6的表达,以探讨CXCL16/CXCR6在蜕膜免疫活性细胞募集中的可能规律.方法收集早孕期蜕膜组织,分离蜕膜基质细胞和免疫细胞,分别采用半定量RT-PCR、免疫细胞化学、流式细胞术分析蜕膜基质细胞CXCL16和CXCR6的表达;流式细胞术分析蜕膜CD56^＋CD16^-NK细胞、CD56^＋CD16^＋NK细胞、NKT细胞、T细胞、γδT细胞、单核细胞CXCR6的表达.结果人早孕蜕膜基质细胞高水平转录趋化因子受体CXCR6,低水平转录其配体CXCL16,但CXCL16和CXCR6在蛋白水平的表达偏低.早孕蜕膜γδT细胞CXCR6阳性率为87.29%;CD14^＋单核细胞CXCR6阳性率为47.71%;NKT细胞CXCR6阳性率为44.14%;T细胞CXCR6表达率为32.91%;而蜕膜两种NK细胞（CD56^＋CD16^-、CD56^＋CD16^＋）几乎不表达CXCR6.结论人γδT细胞、单核细胞、NKT细胞、T细胞可能通过表达趋化因子受体CXCR6被募集到蜕膜局部并驻留,从而参与早孕期母胎界面的免疫调节.     

IFN-γ/IL-4对早孕期蜕膜细胞和绒毛滋养层细胞的体外影响

探讨Th1/Th2型细胞因子对人孕早期绒毛滋养层细胞和蜕膜细胞活性及其内分泌功能的影响。绒毛滋养层细胞和蜕膜细胞活性采用MTT法进行检测 ,绒毛组织分泌的人绒毛膜促性腺激素 (hCG )和蜕膜组织分泌的催乳素 (PRL )均采用放免法 (RIA )进行分析测定。结果 :(1)一定浓度范围的Th1型细胞因子IFN γ (10～ 10 0 0ng/ml)和Th2型细胞因子IL 4 (1～ 10ng/ml)对绒毛滋养层细胞活性及其hCG分泌分别有不同程度的抑制作用 (P <0 0 1)和促进作用 (P <0 0 1) ;(2 )Th1型细胞因子IFN γ低浓度时 (1～ 10ng/ml)对蜕膜细胞活性及PRL分泌有刺激作用 (P <0 0 5 ) ,高浓度时 (10 0～ 10 0 0ng/ml)则有抑制作用 (P <0 0 1) ,Th2型细胞因子IL 4对蜕膜上述功能的调节作用与对绒毛滋养层的调节作用相似。Th1/Th2型细胞因子可能是通过影响绒毛滋养层细胞和蜕膜细胞活性及其内分泌功能而在早期妊娠中起重要的免疫调节作用     

人胸腺基质淋巴细胞生成素(hTSLP)在早孕绒毛组织的表达及意义

目的:探讨人早孕母胎界面表达TSLP及其受体(TSLPR)的特征.方法:使用RT-PCR、Western blot、免疫组织化学法分析正常绒毛组织TSLP/TSLPR的表达;RT-PCR、免疫细胞化学法分析原代培养的滋养细胞TSLP/TSLPR的表达;ELISA检测原代培养的人滋养细胞上清TSLP分泌水平.结果:从转录水平至蛋白水平,绒毛细胞滋养细胞与合体滋养细胞均表达TSLP及TSLPR.结论:TSLP表达于正常早孕绒毛组织,可能与早孕期调节性T细胞的扩增及功能变化相关.     

早孕期人滋养细胞CXCR4/CXCL12的表达及低氧对其的影响

目的：研究人早孕期滋养细胞CXCR4/CXCL12的表达情况及低氧对其表达的调节作用.方法：免疫组织化学法检测早孕期绒毛组织中及原代培养的滋养细胞CXCR4/CXCL12的表达情况;实时定量PCR检测低氧状态下滋养细胞CXCR4 mRNA水平.结果：胎盘绒毛柱、滋养细胞及血管内均有CXCR4/CXCL12表达;低氧培养12、24、48和72小时后滋养细胞CXCR4 mRNA表达增加,显著高于正常氧条件下的表达情况.结论：早孕期胎盘表达CXCR4/CXCL12对于维系正常妊娠的顺利进行可能发挥重要作用;低氧是CXCR4表达的重要调节因子,可能参与了妊娠的病理生理过程.     

白细胞介素25促进早孕期母-胎界面滋养细胞的增殖

探索人早孕绒毛组织和滋养细胞中白细胞介素25(interleukin 25,IL-25)及其受体的表达,以及IL-25对滋养细胞的促增殖作用。免疫组织化学法检测IL-25在正常早孕期绒毛组织的表达;流式细胞术检测IL-25及其受体在正常妊娠和自然流产绒毛组织的滋养细胞中的表达;CCK-8实验分析重组IL-25(rhIL-25)及其中和性抗体对人绒毛膜滋养细胞株HTR-8/SVneo细胞活力的影响。结果显示IL-25在正常绒毛组织滋养细胞中呈阳性表达,自然流产患者绒毛滋养细胞的IL-25及其受体的表达均低于正常妊娠(P0.001);重组IL-25处理后,HTR-8/SVneo细胞活力以时间依赖和剂量依赖方式显著增强(P0.001或P0.000 1);相反,IL-25中和性抗体明显抑制其细胞活力(P0.05或P0.01或P0.001)。结果表明人早孕母-胎界面滋养细胞表达IL-25,以自分泌方式促进自身增殖,IL-25异常低表达可能参与反复自然流产的发病机制。     

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 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.