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1.
树突状细胞体外定向诱导扩增及其分离纯化   总被引:8,自引:0,他引:8  
目的:探讨定向诱导及纯化树突状细胞(DC)的方法,为进一步研究树突状细胞的功能、特性及临床应用提供技术方法。方法:利用免疫磁珠江(MACS)分离纯化脐血CD34细胞,在液体培养体系中加入FL、GM-CSF、TNFα、IL-4ey SCF,培养12d后光镜检测细胞形态及流式细胞仪分析表达标志;利用抗估状细胞单克隆抗体(X-11)及免疫磁珠分离纯化树突状细胞,流式细胞仪检测纯化后DC的纯度。结果:经体  相似文献   

2.
运用流式细胞仪及细胞原位杂交方法测定低氧对培养的新生牛肺动脉平滑肌细胞(PASMC)生长的影响,并探讨血小板源生长因子(PDGF)在其中的作用,结果表明:培养的PASMC的低氧12h时,由G0/G1期进入S斯的细胞数明显增多,为对照组的157.7%(P〈0.05);PDGF可进一步促进低氧状态下PASMC的增殖,S期细胞为对照组的242.3%(P〈0.01),原位杂交结果显示:低氧(12h)条件下  相似文献   

3.
目的:研究抗小鼠胸腺基质细胞(MTSC)Pf18-3单抗识别分子的表达特性。方法:采用流式细胞(FACS)和激光扫描共焦显微镜(CLSM)分析TSC单抗Pf18-3染色特征。结果 :FACS分析发现Pf18-3单抗识别分子可表达于胸腺、胎肝和骨髓等不同来源的基质细胞系和腹腔巨噬细胞表面。新鲜分离的胸腺细胞Pf18-3识别分子表达阴性,但ConA活化可诱导并上调其表达,随活化时间延长表达增高,且主要  相似文献   

4.
IL-3、G-CSF拮抗Ara-c诱导的白血病细胞凋亡的研究   总被引:2,自引:0,他引:2  
目的 研究不同Ara-c浓度时白血病细胞凋亡的情况和G-CSF,IL3+G-CSF对其的拮抗作用,方法:分别用不同剂量Ara-c诱导白血病细胞凋亡,同时加G-CSF,IL-3+G-CSF作为拮抗剂,分G-CSF,IL-3+G-CSF,Ara-c三组。通过形态学,DNA电泳,蛋白电泳免疫印迹法观察白血病细胞的凋亡情况,结果:随着Ara-c剂量的增加,凋亡细胞数增高,G-CSF能拮抗低剂量及常规剂量A  相似文献   

5.
人胸腺上皮细胞(TEc)的体外培养和传代,使研究人TEc的特性和功能成为可能。人TEc体外培养成功的关键是抑制成纤维细胞的生长。体外培养的人TEc除表达角蛋白外,还表达HLA-I类抗原、LFA-3,ICAM-1和CD40等表面抗原;可分泌IL-1,IL-6,IL-8,GM-CSF,G-CSF和M-CSF等多种细胞因子和胸腺素;静息的胸腺细胞与TES的结合由CD2/LFA-3介导,激活的胸腺细胞与TEC的结合由CD2/LFA-3和LFA-1/ICAM-1共同介导;TEC对成熟的胸腺细胞有辅助增殖的作用,对不成熟的胸腺细胞有直接激活作用:胸腺细胞可通过直接作用和分泌细胞因子的间接作用来调节TEC的细胞因子分泌和表面抗原表达。  相似文献   

6.
在建立C3转化酶衰变加速活性检测方法的基础上,成功分离纯化人红细胞膜衰变加速因子(De-cay-acceleratingfactor,DAF,CD55)。以此作为免疫原,制备成功3株抗人DAF单抗DG3、DG9和DA11,分别作用于SCR3、SCR1和SCR4,其中DG3可完全阻断DAF对经典途径C3转化酶的促衰变加速活性。较系统观察了单独或联合使用抗人DAF抗体与抗人CD3单抗对T细胞的活化作用,并证明:①针对SCR3的不同单抗对T细胞活化有不同的作用;②抗人DAF单抗与抗人CD3单抗可协同刺…  相似文献   

7.
用碱性磷酸酶标记的eaeA基因寡核苷酸探针,对37个血清群的221株从肯尼亚5岁以下腹泻患儿粪便中分离的大肠杆菌进行检测,并对eaeA基因阳性菌株进一步做了bfpA和slt基因检测,HEp-2细胞粘附及荧光肌纤蛋白染色(FAS)试验。结果表明,eaeA基因的携带率为19.5%,EPEC、EHEC和其它血清群中eaeA基因的携带率分别为31.6%、66.7%和8.9%,根据bfpA和slt基因检测及HEp-2细胞粘附和FAS试验的结果,可将携带eaeA基因的大肠杆菌分成5种类型。提示eaeA基因的分布不仅限于EPEC和EHEC,而且携带eaeA基因菌株含有多种毒力决定因子。  相似文献   

8.
维持液中小牛血清浓度对LDH法检测NK活性的影响   总被引:4,自引:0,他引:4  
本文对含不同浓度小牛血清维持液中靶细胞的自然释放,最大释放及效应细胞释放进行检测,结果表明,:效应细胞自然释放随FCS为20%时,其释近接近甚至超过量最大释放:FCS低于5%时,基释放趋于稳定。由结果看,无FCS时,各项释放最稳定。  相似文献   

9.
早期作用造血因子对人脐血CD34^+细胞的体外扩增作用   总被引:1,自引:0,他引:1  
为了观察早期作用造血细胞因子SCF、FL、IL-3、IL-6、TPO单独及联合应用,对脐血CD34+细胞的体外扩增作用。我们用吸附单克隆抗体一磁珠分离系统富集人脐血CD34+细胞,在体外液体培养体系中加入不同的细胞因子扩增4周,每周取样计数有核细胞总数及集落形成细胞(CFC)数。结果表明:用磁性细胞分离议富集脐血CD34+细胞纯度为80%~87%;一些细胞因子有明显的协同效应,其联合应用的扩增作用显著高于单因于作用:SCF+FL存在下,IL-3是有效扩增有核细胞总数及CFC的关键因子了;细胞因子SCF+FL+IL-3和SCF+FL+IL-3+IL-6组合对有核细胞总数及CFC均有良好的扩增效应,培养2周时对CFC的扩增倍数分别为38.3±4.4和29.6±2.7倍,可满足成人移植及基因治疗等的需要。  相似文献   

10.
目的:为了探讨一氧化氮合成酶在CD23白血病细胞的表达,以及CD23与NO的关系。方法:采用CRT-PCR和FACS技术及Western-blotting放射自显影方法。结果:发现CD23白血病细胞株可以表达iNOSmRNA和iNOS蛋白,同时CD23可上调iNOS的表达。结论:iNOS可能与人CD23白血病细胞的增殖和凋亡有关。  相似文献   

11.
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.  相似文献   

12.
Renal dysplasia and asplenia in two sibs   总被引:2,自引:0,他引:2  
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.  相似文献   

13.
About 1900, modern food selection and processing caused widespread 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 panhypovitaminosis B, i.e., a mixture of substrate beriberi and substrate pellagra resembling vitamin beriberi and pellagra but exhibiting as even more diverse 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 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.  相似文献   

14.
15.
Newton H 《Medical history》2011,55(2):153-182
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.  相似文献   

16.
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.  相似文献   

17.
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.  相似文献   

18.
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.  相似文献   

19.
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.  相似文献   

20.
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.  相似文献   

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