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1.
目的探究小鼠肝移植模型中移植肝内免疫排斥反应程度和调节性T细胞的数量及功能随时间变化的规律,为揭示机体免疫耐受的建立机制提供理论依据。方法同基因或异基因肝移植术后3、7、14和28 d留取移植肝标本(每组每个时间点n=4),HE染色进行排斥活性指数评定(Banff评分标准),抗CD3、B220和Foxp3抗体免疫组化染色鉴定移植物内T、B淋巴细胞及调节性T细胞的浸润情况,实时荧光定量PCR检测Foxp3 mRNA表达,流式细胞术检测Treg细胞表面CLAT-4的表达。结果异基因肝移植术后2周内发生免疫排斥反应,随后逐渐好转;移植物内浸润的炎性细胞主要为T淋巴细胞,其中Foxp3 mRNA及Foxp3阳性细胞数术后异基因组(15.0±1.3)显著高于同基因组(2.7±1.0)。异基因移植肝内浸润的调节性T细胞表面CLAT-4表达(53%±3%)也显著高于同基因组(13%±2%)(P0.05)。结论异基因肝移植术后免疫排斥反应由调节性T细胞负向调节并达到免疫耐受。  相似文献   

2.
目的研究松果体摘除及补充褪黑素对大鼠胸腺CD4+ CD25+调节性T细胞(Treg细胞)产生、输出及其相关基因叉状头/翅膀状螺旋转录子(Foxp3)表达的影响。方法选用雄性清洁级SD大鼠120只,分为正常组、假手术组、松果体摘除组、松果体摘除+褪黑素低剂量组[腹腔注射7.5mg/(kg·d)]和松果体摘除+褪黑素高剂量组[腹腔注射15mg/(kg·d)],术后4、8周取材。应用流式细胞检测技术、RT-PCR法及免疫组织化学染色法观察并分析松果体摘除及补充外源性褪黑素后胸腺及外周血CD4、CD25双阳性细胞数量及胸腺Foxp3表达的变化。结果松果体摘除术后无论4周或者8周胸腺CD4+ CD25+ Treg细胞数量均无明显变化,补充褪黑素后双阳性细胞数量呈时间、剂量依赖性明显增加;松果体摘除术后4周外周血CD4+ CD25+ Treg细胞数量明显增加,但补充褪黑素后恢复正常,而术后8周各组之间无显著性差异;半定量RT-PCR及免疫组织化学结果显示,松果体摘除后4周大鼠胸腺Foxp3表达水平明显升高,补充高低两种剂量褪黑素后均有所下降并达到正常水平,而松果体摘除后8周各组之间Foxp3的表达无明显差异。结论松果体摘除对胸腺CD4+ CD25+ Treg细胞的产生无明显影响,但导致大鼠胸腺Foxp3的转录和翻译明显增加,胸腺CD4+ CD25+ Treg细胞的输出增加,而补充外源性褪黑素后能逆转上述异常,长时间作用则其影响逐渐消失。  相似文献   

3.
目的 观察佐剂关节炎(AA)大鼠肺功能、调节性T细胞(Treg)及其表面标志物-叉头状转录因子(Foxp3)变化,探讨Treg、Foxp3在RA肺病变中的作用.方法 将30只大鼠随机分为正常组和模型组,每组15只,向模型组大鼠右后足跖皮内注射弗氏完全佐剂0.1 mL致炎,复制成AA模型.致炎30 d后,观察大鼠足跖肿胀度、关节炎指数,采用动物肺功能仪检测大鼠肺功能,流式细胞术检测大鼠外周血Treg表达,RT-PCR、免疫组化、免疫印迹法检测大鼠肺组织Foxp3 mRNA、Foxp3蛋白表达.结果 与正常组比较,AA大鼠组织肿胀度和关节炎指数升高,肺功能参数降低,外周血CD4+ CD25+ Treg、CD4+ CD25+Foxp3+ Treg表达降低,同时,肺组织Foxp3 mRNA和Foxp3蛋白表达水平亦降低(P<0.01或P<0.05).结论 AA大鼠在发生关节炎症的同时,其肺功能水平降低,Treg、Foxp3表达降低,免疫平衡失调.  相似文献   

4.
目的:研究褪黑素在抑制肿瘤过程中对荷胃癌小鼠胸腺及脾CD4+CD25+调节性T(Treg)细胞表达变化的影响.方法:培养小鼠前胃癌细胞株(MFC),采用小鼠荷胃癌模型.用褪黑素干预1周后,测量小鼠胸腺、脾质量;并用流式细胞仪检测胸腺、脾Treg细胞;用实时荧光定量PCR和免疫印迹法检测胸腺、脾叉头型转录因子3 (Foxp3)的表达.结果:与正常对照组相比,小鼠胸腺、脾质量各组之间差异无统计学意义.在褪黑素抗肿瘤过程中,与正常对照组相比,荷瘤空白对照组小鼠胸腺与脾的Treg细胞均上升,褪黑素干预后降至正常水平;褪黑素还能使小鼠胸腺Foxp3 mRNA表达下降,脾Foxp3 mRNA反而上调;而小鼠胸腺、脾scurfin蛋白均明显下降.结论:褪黑素能下调荷胃癌小鼠胸腺、脾的CD4+CD25+Treg细胞及特异性转录蛋白scurfin及胸腺Foxp3 mRNA,使脾Foxp3 mRNA反而上调.褪黑素抗胃癌作用与免疫器官中CD4+CD25+Treg细胞免疫调节有关.  相似文献   

5.
目的:研究雷帕霉素(Rapamycin,RAPA)和环孢素A(cyclosperin A,CsA)体内处理对同种移植小鼠急性排斥过程中TLR5及Foxp3表达的影响.方法:建立同种皮肤移植模型,术后给予RAPA或CsA,1次/d,连续14 d,同时设生理盐水对照组.每日观察移植物存活状况.术后选取1、7、10、14、21 d共5个时间点,提取受体小鼠脾细胞,RT-PCR检测TLR5及Foxp3 mRNA表达,探讨不同处理组基因表达与移植物生存的相关性.体外实验利用鞭毛蛋白(flagellin)联合RAPA和CsA处理正常小鼠T细胞6 h,RT-PCR检测TLR5表达.结果:RAPA和CsA可明显延长小鼠同种移植皮片存活期.RAPA可促进脾细胞TLR5 mRNA表达升高,停药后的第21天仍明显高于CsA组和对照组(P<0.05);CsA组在第7、10天有显著升高,第21天降低(P<0.05);3组中最高表达的时间点为移植后第10天,此时正值排斥高峰期.RAPA可引起Foxp3 mRNA的表达升高(P<0.05),停药后仍维持较高水平;CsA组仅在移植后第7、10天短暂升高,第14天低于对照组(P<0.05).移植后1、7、10、21 d 3组的TLR5与Foxp3 mRNA变化趋势正相关.体外实验中,RAPA或CsA与flagellin联合使用,可促进TLR5的表达,以前者的作用更为明显.结论:RAPA和CsA在同种移植急性排斥高峰期可引起TLR5和Foxp3表达的协同升高,且RAPA的作用更加明显和持久,鞭毛蛋白体外可以增强这种作用效果.  相似文献   

6.
以5mg/kg浓度LPS腹腔注射C57BL/6小鼠建立内毒素耐受模型,观察注射72h及8d后脾脏大小、重量及总细胞数;HE染色观察脾脏病理学变化;FACS检测CD4+Foxp3+Treg细胞比例并计算其细胞总数;同时检测CD4+Foxp3+Treg细胞功能相关膜分子CTLA-4的相对表达;Ki-67染色法检测CD4+Foxp3+Treg细胞的增殖情况;PI染色法检测CD4+Foxp3+Treg细胞的凋亡情况;Real-time PCR技术检测脾脏中TGF-β的相对表达。结果发现与对照组相比,LPS注射72h后脾脏大小、重量及细胞总数显著增加(P0.05),且发生病理学改变;脾脏中CD4+Foxp3+Treg细胞比例和数量显著减少(P0.05),CTLA-4表达水平显著降低(P0.05);同时,增殖比例明显减少,凋亡比例增加(P0.05);最后,脾脏中TGF-β的相对表达水平显著减少。LPS注射8d后脾脏大小、重量及细胞总数均与对照组相比无差异(P0.05),且脾脏组织结构未见改变;脾脏中CD4+Foxp3+Treg细胞比例及数量仍减少(P0.05),而其表达CTLA-4水平增加(P0.05),且凋亡和增殖比例无明显差异(P0.05);最后,脾脏组织中TGF-β的相比表达显著增加。结果表明,内毒素耐受状态下,CD4+Foxp3+Treg细胞比例发生明显改变,可能与细胞凋亡和增殖变化有关。  相似文献   

7.
 目的:观察针刺对胚胎着床障碍大鼠CD4+CD25+Foxp3+Treg细胞的影响。方法:144只孕鼠随机分为对照组(N)、米非司酮组(M)、米非司酮+针刺组(A)和米非司酮+黄体酮组(W),每组随机再分为6 d组、8 d组和10 d组。其中M组、A组和W组妊娠1 d给予米非司酮-麻油溶液造模,N组则给予等量麻油溶液。同时,每天下午A组固定并行针刺三阴交和后三里,N组和M组仅固定,W组肌肉注射黄体酮,直到处死当天结束。统计孕鼠胚胎着床数,用流式细胞术检测外周血CD4+CD25+Foxp3+Treg细胞和子宫内膜的CD4+Foxp3+Treg细胞的比例,Western blotting和real-time PCR测定着床点的子宫内膜Foxp3的表达。结果:与N组相比,M组的着床胚胎数、外周血CD4+CD25+Foxp3+Treg细胞和子宫内膜CD4+Foxp3+Treg细胞的比例、着床点子宫内膜Foxp3蛋白和mRNA的表达均明显下降(P<005);与M组相比,A组和W组的着床胚胎数、外周血CD4+CD25+Foxp3+Treg细胞和子宫内膜CD4+Foxp3+Treg细胞的比例以及着床点子宫内膜Foxp3蛋白和mRNA的表达均有不同程度地升高。结论: 针刺改善胚胎着床障碍大鼠的胚胎着床可能与CD4+CD25+Foxp3+Treg细胞的调节密切相关。  相似文献   

8.
目的:研究哮喘小鼠CD4+CD25+Foxp3+调节T(Treg)细胞和Foxp3蛋白的变化,探讨淫羊藿苷对哮喘干预机制。方法:将BALB/c小鼠32只随机分为正常对照组、哮喘模型组、淫羊藿苷组、地塞米松阳性对照组,每组8只。哮喘模型制备用卵白蛋白(OVA)致敏大鼠并雾化吸入刺激,治疗组采用相应药物灌胃给药,对照组用等量生理盐水代替。最后一次激发24小时后,采用Buxco肺功能仪有创法检测小鼠气道反应性;HE染色评价观察小鼠肺组织病理形态学改变;流式细胞术检测脾脏CD4+CD25+Foxp3+Treg细胞比例及Foxp3蛋白表达量;免疫印迹法测定肺组织Foxp3蛋白表达量;ELISA法测定血清及肺泡灌洗液(BALF)IL-10水平。结果:与正常对照组比较:模型组气道阻力及气道炎症指数显著增加(P<0.05);脾Treg细胞比例无明显变化(P>0.05);脾Foxp3蛋白表达显著下降(P<0.05),肺组织Foxp3蛋白表达显著升高(P<0.05),外周血IL-10水平显著下降(P<0.05),BALF中IL-10无显著差异(P>0.05);与模型组比较:淫羊藿苷组气道阻力和气道炎症明显减轻(P<0.05);脾Treg细胞比例和脾Foxp3蛋白表达水平无显著变化(P>0.05),但淫羊藿苷可进一步上调哮喘小鼠肺组织Foxp3蛋白表达量(P<0.05),并增加外周血IL-10水平(P<0.05)。结论:哮喘小鼠脾脏CD4+CD25+Foxp3+Treg细胞Foxp3蛋白表达水平下降,淫羊藿苷可显著改善哮喘小鼠气道炎症和气道高反应,可能与其上调肺组织Foxp3蛋白表达和增加外周血IL-10水平相关。  相似文献   

9.
目的观察慢性阻塞性肺疾病(COPD)大鼠调节性T细胞(Treg)、叉头状转录因子3(Foxp3)、T-bet、GATA连接蛋白3(GATA3)的变化。方法将30只大鼠随机分为对照组、模型组,每组15只。模型组大鼠采用烟熏加脂多糖(LPS)气管滴入方法建立COPD模型。模型复制成功第28天,采用ELISA检测大鼠血清、支气管肺泡灌洗液(BALF)白介素4(IL-4)、γ干扰素(IFN-γ)水平,流式细胞术检测外周血Treg表达,分别采用逆转录PCR及Western blot法检测肺组织Foxp3、T-bet、GATA3mRNA和蛋白表达。结果模型组大鼠肺泡腔及肺间质内大量炎性细胞浸润,肺组织结构破坏。与对照组比较,模型组大鼠血清IFN-γ表达升高(P0.01),IL-4、CD4+CD25+Treg、CD4+CD25+Foxp3+Treg降低(P0.05);模型组Foxp3、GATA-3mRNA、蛋白表达降低,T-bet mRNA和蛋白升高(P0.01)。相关性分析显示,T-bet、GATA-3、Foxp3基因和蛋白表达与IL-4、IFN-γ分泌相关(P0.05)。结论 COPD炎症反应与T-bet、GATA-3、Foxp3表达失衡有一定关系。  相似文献   

10.
目的:免疫炎症反应在急性缺血性脑卒中的病理生理过程中发挥着重要的作用。具有促炎作用的辅助性T细胞17(Th17)及维持免疫耐受的调节性T细胞(Treg)是体内重要的2种免疫细胞。Th17/Treg细胞平衡是机体维持正常免疫的基础。本研究探讨急性缺血性脑卒中大鼠脑组织中Th17/Treg的变化。方法:采用线栓法制备SD大鼠急性大脑中动脉闭塞(middle cerebral artery occlusion,MCAO)模型,以假手术组作为对照组。在MCAO术后3 d利用TTC染色观察各组大鼠脑梗死体积;采用ELISA测定脑组织中白细胞介素17A(IL-17A)和IL-10蛋白的含量;采用RT-qPCR测定脑组织中IL-17、IL-10、Foxp3和RORγt的mRNA表达水平;采用流式细胞术测定脑组织中Th17细胞和Treg细胞的比例变化。结果:与假手术组相比,MCAO组大鼠脑组织中IL-17A的含量增加,IL-10的含量减少(P0.05);RORγt和IL-17的mRNA表达水平上调(P0.05),Foxp3和IL-10的mRNA表达水平下调(P0.05);脑组织Th17细胞增多,Treg细胞明显减少(P0.05),Th17/Treg比值升高(P0.05)。结论:急性缺血性脑卒中大鼠脑组织Th17细胞增多,Treg细胞减少,表明脑梗死后大鼠脑组织中Th17/Treg的平衡被破坏,免疫炎症反应被激活。  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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