负载HSP60树突状细胞对小鼠动脉粥样斑块影响研究

目的： 探讨负载热休克蛋白60(HSP60)树突状细胞(DC)接种对ApoE-null小鼠粥样硬化斑块的影响。 方法：小鼠髓源性DC体外培养成熟，分别用磷酸缓冲液（PBS）、卵清蛋白（OVA）和HSP60处理，体外检测各组DC的功能；同系小鼠予以高脂饮食16周形成斑块，各组DC用荧光物质标记后，分别经皮接种3次，48 h 后取主动脉HE染色及荧光观察，测血清IL-10、IFN-γ浓度。 结果：体外HSP60及OVA可促进DC表达CD86，而PBS则无此效应。接种小鼠后，HSP60-DC和OVA-DC组血清IFN-γ较PBS-DC组高；而IL-10无明显差别；IFN-γ/IL-10 比值增高。HSP60-DC促使主动脉粥样斑块炎性细胞浸润明显增加，斑块趋于不稳定；OVA-DC则对斑块无显著效应。 结论：HSP60负载的DC可以特异性刺激主动脉粥样斑块炎性反应，诱导炎性细胞因子释放，引起免疫偏移。     

阿司匹林对树突状细胞成熟及免疫功能的体外研究

研究阿司匹林在体外对小鼠树突状细胞(mDC)成熟及免疫功能的影响。在骨髓来源未成熟树突状细胞培养过程中,加入不同剂量(1mmol/L、2mmol/L)的阿司匹林,观察DC形态。流式细胞仪检测各组mDC表面共刺激分子CD86、CD80的表达;台盼蓝染色测细胞活力;混合淋巴细胞反应(MLR)检测mDC对T淋巴细胞刺激能力;ELISA法检测MLR上清液中细胞因子。与对照组比,阿司匹林处理的DC(aspirin-DC)表面CD80、CD86表达明显降低(P<0.01);对T淋巴细胞刺激能力减弱;MLR上清液中炎性因子(TNF-α、IL-1β)明显减少(P<0.01)。结果:在体外培养过程中给予阿司匹林干预能够抑制mDC的成熟及功能,呈剂量依赖性;阿司匹林对DC功能的抑制可能是阿司匹林抑制炎症反应,治疗冠心病的机制之一。     

CD40配基化调节小鼠树突状细胞上B7-H3分子表达的研究

目的 探讨CD40配基化对小鼠骨髓来源树突状细胞上B7-H3分子表达的调节作用及其生物学意义。方法 采用GM-CSF和IL-4联合方案体外诱导小鼠髓系DC，并利用mCD40-CHO和TNF-α分别刺激凋亡肿瘤细胞负载的Dc制备成熟DC；采用间接免疫荧光标记法检测成熟Dc上B7-H3分子的表达；RT-PCR检测B7-H3 mRNA转录水平；混合淋巴细胞反应（MLR）和B7-H3单抗阻断实验分析CD40配基化的DC表面B7-H3分子在T细胞活化中的作用；^3H-TdR掺入试验检测DC对T淋巴细胞的促增殖效应；ELISA测定各组MLR反应和DC培养上清中IFN-γ分泌水平。结果 B7-H3分子在DC不同分化发育阶段均有表达，CD40配基化能显著上调凋亡肿瘤细胞负载的DC中B7-H3表达，TNF-α激发的DC弱表达（P〈0．05）；阻断CD40配基化的DC上B7-H3分子能抑制T细胞增殖和IFN-γ分泌；CD40配基化促进凋亡肿瘤细胞负载的DC分泌IFN-γ量也明显高于TNF-α组（P〈0．05）。结论 体外CD0配基化DC的B7-H3分子上调性表达有助于其刺激T细胞增殖和IFN-γ的产生。     

HBsAg重组腺病毒转染的树突状细胞诱导BALB/c小鼠抗HBV免疫的研究

目的 研究HBsAg重组腺病毒转染的小鼠树突状细胞（DC）体内外免疫刺激活性和诱导小鼠抗HBV免疫的特点。方法 BALB／c小鼠的骨髓细胞体外扩增为DC，转染HBsAg重组腺病毒Ad-S或被HBsAg蛋白冲击后，流式细胞术分析DC的表型，混合淋巴细胞反应（MLR）检测DC的刺激活性；或与pcDNA3．1（＋）-S质粒分别免疫小鼠后，LDH法测定脾细胞CTL活性，放免法检测血清抗．HBs；流式细胞术分析体外自体MLR中及免疫后小鼠脾脏T细胞内细胞因子。结果 DC／Ad-S、DC／HBsAg和各对照组DC之间的表型和MLR差异无统计学意义，并均刺激TH和Tc分泌IFN-γ。DC／Ad-S免疫后2周能诱导比DNA疫苗更强产生IFN-γ的TH（P〈0．05），以及比DC／HBsAg和DNA疫苗更强分泌IFN-γ的Tc（均P〈0．01）和特异性CTL（P〈0．05，P〈0．01）。但DC／Ad-S和DC／HBsAg诱导的CTL反应及Tc分泌IFN-γ在免疫后4周均明显减弱，且诱导抗．HBs作用弱于DNA疫苗。结论 HBsAg重组腺病毒转染的DC比HBsAg冲击的DC及DNA疫苗能诱导更强的TH1/Tcl（Ⅰ型）细胞免疫和特异性CTL反应，是诱导抗HBV细胞免疫的有效刺激细胞。     

香加皮羽扇豆烷乙酸酯(CPLA)对树突状细胞分化成熟的影响

目的：探讨香加皮羽扇豆烷乙酸酯（CPLA）对人外周血单个核细胞（PBMC）来源的树突状细胞（DC）在体外分化成熟及免疫活性的影响。方法：从人外周血分离单个核细胞，与细胞因子GM—CSF、IL-4共培养，于第5天加入DC的促成熟刺激剂TNF-α（阳性对照组）或CPLA。倒置显微镜和透射电镜下观察DC的形态；应用流式细胞术检测成熟DC的表面标志CD1a、CD83、CD80和CD86的表达情况；用ELISA检测DC培养上清中IL-12和IFN-γ的含量；用MTT法测定DC刺激T细胞增殖的能力。结果：培养10d后，经CPLA刺激的PBMC呈现出典型DC的形态学特征；成熟DC的特征性表面分子CD1a、CD83、CD80和CD86表达水平均明显上调（P〈0．05）；细胞培养上清中IL-12和IFN-γ含量明显增高（P〈0．05）；刺激T细胞增殖的能力明显增强（P〈0．05）。结论：CPLA可诱导PBMC来源的DC分化成熟，并可促进其细胞因子的分泌，增强DC的免疫调节活性。     

慢性乙型肝炎患者树突状细胞体外功能恢复及诱导T细胞反应研究

目的 研究慢性乙型肝炎(CHB)患者外周血树突状细胞(DC)和T细胞体外功能,并尝试恢复功能的方法.方法 12例CHB患者作为研究对象,10例健康人作为正常对照研究.对外周血DC进行表型分析和混合淋巴细胞反应(MLR)测定,并分别用HBsAg或HBcAg作为抗原负载DC,后用ELIspot法检测产生IFN-γ的细胞频数.结果 CHB患者外周血DC共刺激分子水平表达降低,刺激MLR能力不足.经过体外补充促成熟细胞因子,DC缺陷能够得到部分纠正.成熟DC较不成熟Dc刺激MLR和诱导产生IFN-γ的T细胞能力强,HBcAS比HBsAS有更强的免疫原性.结论 慢性CHB患者外周血DC功能缺陷,能够通过补充促成熟因子纠正.成熟DC比不成熟DC显示出更强的MLR反应和诱导IFN-γ产生细胞的能力,这提示HBV蛋白负载DC作为治疗性疫苗具有可行性.     

IL-10诱导小鼠树突状细胞耐受的分子机制

目的：研究白细胞介素-10 (interleukin-10，IL-10)诱导小鼠来源的树突状细胞(DC)耐受及其与配对免疫球蛋白样受体（PIR-A/B）的关系。方法: 以IL-10（20 μg/L）诱导小鼠来源的树突状细胞系（DC2.4）6 d，即IL-10-DC组，脂多糖（LPS）刺激其48 h为成熟DC2.4细胞（LPS-DC），体外化学合成特异性针对PIR-B的小干扰RNA片段，以脂质体2 000转染IL-10组（Si-DC组）。分别应用半定量RT-PCR和流式细胞仪（FCM）检测DC2.4、IL-10组、LPS组及Si-DC组细胞PIR-A/B的表达。以［3H］-TdR标记法检测上述各组细胞刺激同种异体淋巴细胞的增殖反应(MLR)，ELISA方法测混合培养上清中IFN-γ的水平变化。结果: RT- PCR结果表明，IL-10诱导PIR-B表达升高、PIR-A表达下降，LPS则下调PIR-B、上调PIR-A的表达。FCM检测IL-10组和LPS组的PIR-A/B胞外区PIR表达均升高，且前者明显高于后者。同正常DC2.4和LPS组相比，IL-10可抑制MLR，小干扰RNA沉默PIR-B表达可增强MLR，伴随MLR反应上清中IFN-γ的水平升高。结论: IL-10诱导DC高度表达免疫抑制性受体PIR-B，使其获得耐受，上调PIR-B的表达是IL-10诱导DC耐受的分子机制之一。     

阻断CD40-CD40L共刺激途径对T细胞表型及细胞因子分泌的作用研究

目的探讨应用抗CD40L单克隆抗体阻断CD40-CD40L共刺激途径后对T细胞表型及其分泌的细胞因子的影响,为体外阻断该共刺激途径诱导T细胞对异体移植抗原的免疫耐受提供实验依据.方法供鼠(C57BL/6H-2b)脾T细胞作为反应细胞,受鼠(BALB/CH-2d)脾细胞作为刺激细胞,设单抗组(加抗CD40L单抗)和对照组(不加单抗),初次混合淋巴细胞培养(MLR)7天,在不同时间点采用3H-TdR掺入法检测细胞增殖率,以ELISA法测定培养上清液中IFN-γ、IL-2、IL-4、IL-10等的水平,第5天采用流式细胞仪检测CD4+T和CD8+T细胞上CD25、CD69、CD40L和CD45RA的表达.再次MLR 5天,第1、3、5天采用3H-TdR掺入法测定细胞的增殖情况和ELISA法测定培养上清液中的上述细胞因子的水平.结果初次和再次MLR结果均显示,单抗组细胞增殖反应率明显低于对照组.初次MLR单抗组中CD4+T和CD8+T细胞比例明显低于对照组(P＜0.05);单抗组中CD4+CD25+T、CD4+CD69+T、CD8+CD25+T、 CD4+CD40L+T和CD8+CD69+T细胞比例明显低于对照组(P＜0.05),而CD8+CD40L+T和CD4+CD45RA+T细胞的比例与对照组相比无明显差异(P＞0.05).初次MLR中单抗组和对照组培养上清中IL-4和IL-10几乎无法测出,而单抗组培养上清中IFN-γ和IL-2的水平均明显低于对照组(P＜0.01);再次MLR后培养上清中单抗组IFN-γ、IL-2和IL-4和IL-10的分泌水平明显低于对照组(P＜0.05),但处于低水平,仍明显低于对照组.结论在体外MLR体系中,应用抗CD40L单抗孵育供鼠脾T细胞,可同时作用于CD4+T和CD8+T细胞,使CD40L+,CD25+和CD69+表达下降,引起T细胞早期的活化和成熟障碍,T细胞增殖能力减低,抑制了Th1类细胞因子IFN-γ和IL-2及Th2类细胞因子IL-4和IL-10的分泌水平,可诱导供者T细胞免疫耐受.     

未成熟树突状细胞对T细胞活化及其IFN-γ和IL-12表达的影响

目的:探讨未成熟树突状细胞(DC)对T细胞活化及其IFN-γ和IL-12表达的影响。方法:通过诱导Lewis大鼠骨髓前体细胞,获取未成熟(iDC)和成熟(mDC)两种状态的DC;将两种DC用乙酰胆碱受体(AChR)负载后进行T细胞重复刺激、交叉刺激和无关抗原刺激试验,观察T细胞增殖情况并测定T细胞IFN-γ、IL-12的表达水平。结果:(1)AChR负载的iDC可明显抑制T细胞增殖,且这种被抑制的T细胞对AChR负载的mDC的交叉刺激也不引起增殖,但对无关抗原OVA负载的mDC的刺激可产生明显增殖。(2)与mDC组比较,AChR负载的iDC初次和重复刺激均明显抑制T细胞IFN-γ和IL-12的表达。结论:iDC可诱导抗原特异性T细胞耐受,耐受机制可能与Th1细胞因子IFN-γ和IL-12的抑制有关。     

脐带血树突状细胞的免疫功能研究

目的： 探讨脐带血树突状细胞(DC)的免疫功能状况。 方法： 以CD34-Lin- HLA-DR+细胞群设门4色荧光分析方法检测20例脐带血和20例外周血DC前体细胞亚群pDC1/pDC2（CD11c+CD123-/CD11c-CD123+）比值；酶联免疫吸附法检测其血浆相关细胞因子IL-12p40、IL-10、IFN-γ、IL-4水平。 结果： 脐带血、成人外周血pDC1/pDC2比值无显著差异（P>0.05）；脐带血血浆IL-12p40水平显著高于成人外周血（P<0.01），两者IL-10、IFN-γ、IL-4水平无显著差异（P>0.05）。 结论： 脐带血DC功能发育比较完善。     

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.     

The universal muscular chain reaction,muscle spasm,Torsions, ruptures and extravasations. Chameleons of pathology and some manifestations of simple muscular disorders

Most bodily functions require the coordinated actions of complementary and supplementary paired muscle groups. Where this essential muscular cooperation is lacking, hollow organs may burst and others become literally screwed up, giving rise to many similar spastic diseases such as Torticollis, Twisted ovarian cyst, Torsion of the Testis, Volvulus of the intestines, Varicose Veins, Megacolon, Aortamegaly, Scoliosis, Erb's Palsy, Peyronie's Disease, Main-en-Griffe, Undescended Foot (Pes Cavus), Talipes, Strabismus. Spasm is “panenepidemic” and unclassified examples of Torsion Dystonia and Dyskinesia really are as common as debt and taxes.     

'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.     

Hypo- und Hypermagnesämie aus kardiologischer Sicht

Zusammenfassung Eine Reihe pathologischer Zustände bedingen Magnesiummangel. Zustände mit Hypermagnesämie sind ebenfalls bekannt, doch wesentlich seltener. Für den Kardiologen beachtenswert ist, daß unter Therapie mit bestimmten Diuretica bei Herzinsuffizienz, bei Herzinfarkt, Kardiomyopathie, Digitalisintoxikation und bestimmten Herzrhythmusstörungen Hypomagnesämie beobachtet wurde. Leider kann in der klinischen Routine nur ein extracelluläres Magnesiumdefizit durch Serumbestimmungen gemessen werden; über Magnesiummangel einzelner Organe kann nichts ausgesagt werden. Hinweise für Magnesiummangel geben aber neben der Messung des Serumspiegels Anamnese, klinischer Befund, bestimmte EKG-Veränderungen wie auch evtl. Hypokalämie, ein Zustand, bei dem sich oft — besonders bei Aldosteronismus — parallele Veränderungen zeigten.Tierexperimente deuten darauf hin, daß infarktähnliche Läsionen unter Magnesiummangel entstehen, doch ob Herzinfarkt beim Menschen durch Magnesiummangel ausgelöst werden kann, ist noch ungeklärt. In Leichenherzen zeigte sich im Infarktgebiet neben Calciumakkumulation signifikanter Magnesiumverlust, wobei unklar blieb, ob sich Ursache oder Folge des Infarktes widerspiegelten. Falls ein ursächlicher Zusammenhang besteht, ist er im Myokardstoffwechsel selbst zu suchen, wie bei der Alkoholkardiomyopathie, wo myokardialer Magnesiummangel zumindest als pathogenetischer Teilfaktor anerkannt wird. Andererseits versucht man aber auch Beziehungen zwischen Atherosklerose, Blutgerinnung und Hypomagnesämie herzustellen, in der Meinung, daß Magnesiummangel auch über den coronaren Pathomechanismus des Herzinfarktes wirken könnte. Sicher scheint, daß gewisse EKG-Veränderungen und Herzrhythmusstörungen durch einen irritierten Magnesiumhaushalt bedingt sein können, da sie bei Gabe bzw. Entzug von Magnesium verschwinden. Daß Magnesiummangel die Glykosidtoleranz verringert, wird tierexperimentell bestätigt. Unter Hypomagnesämie bewirkt Acetylstrophanthidin eher und länger Rhythmusstörungen als ohne, außerdem lassen diese sich durch Magnesiumgaben eliminieren. Da in gewissen Fällen spontane und digitalisinduzierte Herzrythmusstörungen durch Magnesiuminjektionen beseitigt wurden, scheint Magnesium als Therapeuticum angebracht. Einsatz verschiedener Magnesiumsalze bei Angina pectoris, degenerativen Herzerkrankungen und Herzinsuffizienz ohne geprüften und offensichtlich gestörten Magnesiumhaushalt ist fragwürdig, weil keine eindeutigen klinischen Erfolgsbeweise vorliegen. Immerhin mag es aber larvierte, durch Serumbestimmungen nicht erfaßbare Mangelzustände geben. Allgemein erscheint es aus kardiologischer Sicht ratsam, den Magnesiumhaushalt zu überwachen und in entsprechenden Fällen auszugleichen, um möglichen Myokardläsionen oder fatalen Herzrhythmusstörungen entgegenzuwirken.     

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.