自体DC治疗对大肠癌术后患者免疫功能的影响

目的探讨热休克凋亡的人自体大肠癌细胞致敏的树突状细胞（DC）疫苗对大肠癌术后免疫功能的影响。方法从大肠癌患者外周血单核细胞中诱导DC，并用重组人粒细胞-噬细胞集落刺激因子（rhGM-CSF）和白介素4（IL-4）刺激活化，经热休克凋亡自体大肠癌细胞致敏制备DC疫苗。将26例大肠癌术后患者随机分为DC疫苗治疗组13例，化疗对照组13例。对两组病例治疗前后免疫功能、临床疗效进行观察比较。结果DC疫苗治疗后外周血CD3^＋、CD4^＋／CD8^＋及NK细胞比率较治疗前明显升高，P〈0．05，且明显高于对照组化疗后的CD3^＋、CD4^＋／CD8^＋及NK细胞比率，P〈0．05；DC疫苗治疗后血清IL-2／IL-12／IFN-r水平较治疗前明显升高，P〈0．05，且明显高于对照组化疗组，P〈0．05。结论大肠癌术后行热休克凋亡自体大肠癌细胞致敏的DC疫苗治疗，可改善患者的细胞免疫水平，且不良反应少，值得推广应用。     

SLC基因修饰树突状细胞及不同免疫方式抗肿瘤作用探讨

目的：应用次级淋巴组织趋化因子（SLC）成熟肽基因／重组腺相关病毒（rAAV—SLC）转染树突状细胞（DC），探讨一种基因修饰DC的新方法及其抗肿瘤的生物学活性。方法：体外构建rAAV-SLC，按MOI为10、50、100与腺病毒（Ad2）协同转染小鼠骨髓来源成熟和未成熟DC，GFP作为成功转染的示踪蛋白。RT—PCR在mRNA水平、ELISA在蛋白水平检测SLC表达，Transwell检测转染DC的培养上清对T细胞的趋化作用。瘤苗的在体效应通过C57BL／6J小鼠肿瘤模型验证。皮下接种Hepal-6肿瘤细胞后成瘤小鼠分成3组：①生理盐水对照组，瘤内或足垫注射NS；②单纯DC对照组，瘤内或足垫注射未修饰DC；③SLC基因修饰DC组，瘤内或足垫注射SLC基因修饰的DC。每隔7天注射1次，观察各组小鼠肿瘤的生长情况。4周后分离瘤体，用荧光免疫组化检测瘤体内CD4^＋T细胞、CD8^＋T细胞浸润情况。结果：rAAV—SLC在MOI为100时能成功转染DC，未成熟DC的转染效率显著高于成熟DC，并产生对T细胞有明显趋化生物学活性的SLC。动物实验表明，瘤内注射SLC修饰后DC，肿瘤大小与对照组有明显差异，免疫组化显示瘤体内有较多的CD4^＋T细胞、CD8^＋T细胞浸润。足垫注射SLC修饰的DC与对照组相比，抗肿瘤作用无显著差异。结论：rAAV—SLC在Ad2的辅助下能有效转染未成熟DC并能表达具有趋化生物学活性的SLC。SLC基因修饰的树突状细胞瘤内注射以其对CD4^＋T细胞、CD8^＋T细胞的吸引作用而发挥了明显的抗肿瘤作用，同时也进一步证实用树突状细胞进行肿瘤生物免疫治疗时，不同免疫方式对疗效有明显影响。     

氟达拉滨联合瘤苗抗肿瘤生物治疗作用的实验研究

目的探讨氟达拉滨对小鼠CD4＋CD25＋Treg细胞的影响，同时研究其抗肿瘤作用。方法氟达拉滨或生理盐水分别腹腔注射10只小鼠，用流式细胞术检测CD4＋CD25＋Treg细胞相对量。氟达拉滨或生理盐水分别腹腔注射10只小鼠，4d后用丝裂霉素灭活的肿瘤细胞免疫小鼠，观察小鼠抗肿瘤的能力（观察肿瘤发生率和出瘤时间）；用乳酸脱氢酶杀伤实验进一步验证氟达拉滨可增强CTL的杀伤活性。结果氟达拉滨组CD4＋CD25＋Treg细胞占淋巴细胞百分比为（1．150±0．256）％，对照组为（1．488±0．270）％，氟达拉滨组明显低于对照组（P〈0．05）；氟达拉滨＋接种瘤苗组、生理盐水＋接种瘤苗组、氟达拉滨＋未接种瘤苗组和生理盐水＋未接种瘤苗组小鼠第13天肿瘤发生率分别为10％、30％、50％和70％，两两间差异均有统计学意义（P〈0．05）；氟达拉滨＋接种瘤苗组、生理盐水＋接种瘤苗组、氟达拉滨＋未接种瘤苗组和生理盐水＋未接种瘤苗组小鼠分别在接种后第13天、第10天、第8天和第5天发现出瘤现象；对照组的肿瘤生长曲线较为陡直，氟达拉滨组的生长曲线较为平缓。氟达拉滨联合瘤苗接种组和单纯瘤苗接种CTL的活性分别为24．425±13．544和17．575±10．260，两者间差异有统计学意义（P〈0．05）。结论低剂量氟达拉滨降低CD4＋CD25＋Treg细胞，使其抗肿瘤免疫作用增强，联合接种灭活瘤苗进一步增强抵抗肿瘤攻击的能力。     

TLR4在HSP70_(EC-TCV)冲激的DC成熟过程中的作用

观察TLR4在Hca-F榄香烯复合瘤苗来源的HSP70(HSP70EC-TCV)冲激处理的小鼠骨髓来源的DC成熟过程中的作用。用rmGM-CSF和rmIL-4诱导小鼠骨髓来源的DC,以HSP70EC-TCV或加入抗TLR4抗体30 min后再用HSP70EC-TCV冲激处理DC,流式细胞仪检测DC的CD40和CD86表达,ELISA法检测DC上清中IL-12和T细胞上清中IL-2浓度,MTT法检测T细胞对Hca-F细胞的杀伤率。结果表明,抗TLR4抗体对DC的CD40和CD86表达无明显影响,但对HSP70EC-TCV诱导DC分泌IL-12和进而致敏的T细胞分泌IL-2及产生杀瘤功能有明显的抑制作用。TLR4信号通路参与HSP70EC-TCV诱导DC成熟过程。     

MAGE-1与IL-18共表达DNA疫苗在小鼠体内诱导特异性免疫应答能力的研究

目的：观察已构建的peIL-18-MAGE共表达质粒疫苗接种小鼠所引起的免疫应答情况。方法：质粒大量制备肌注小鼠，每隔10天加强免疫1次，共免疫3次。末次免疫后7天，收集血清及脾细胞，流式细胞术检测小鼠T细胞亚群情况及血清中抗MAGE-1多抗的产生，MTT法检测小鼠CTL杀伤活性。结果：peIL-18、pcMAGE-1、peIL-18＋pcMAGE-1及peIL-18-MAGE免疫组CD3^＋、CD4^＋、CD8^＋、CD4^＋／CD8^＋及MAGE-1抗体的产生均高于peDNA3免疫组（P〈0．05），且peIL-18-MAGE共表达免疫组高于peIL-18＋peMAGE-1共同注射组（P〈0．05）。pdL-18-MAGE免疫组对SMMC-7721的杀伤活性最高。结论：pelL-18-MAGE共表达基因疫苗接种小鼠与其它实验小鼠相比，可以有效产生更好的特异性免疫应答。     

糖基化修饰的DC疫苗特异性抗骨髓瘤活性的体外研究

目的：探讨经过糖基化修饰改造过的肿瘤相关糖抗原冲击树突状细胞（DC）制备的DC疫苗特异性抗骨髓瘤作用。方法：采用化学方法及肿瘤细胞的生物工程法使骨髓瘤细胞表达新肿瘤相关抗原N-丙酰多聚唾液酸（NPrPSA）；在无血清培养条件下，采用GM—CSF／IFN-α及TNF-α从MM患者外周血单核细胞诱导培养DC。然后用表达新抗原的肿瘤细胞冲击DC制备DC疫苗，将其刺激T细胞后，通过LDH释放法观察其对骨髓瘤细胞的杀伤效力，并采用ELISA法测定了T细胞分泌细胞因子IFN-γ的能力。结果：糖基化修饰的CD138^＋Npr—DC组LDH释放率较CD138^＋未修饰DC组明显增高（P〈0．01）。CD138^＋Npr—DC组T细胞分泌IFN-γ与CD138^＋未修饰DC组相比亦明显增加（P〈0．05），而CD138^＋Npr—DC组与CD138^-未修饰DC组之间无统计学意义（P〉0．05）。结论：糖基化修饰的DC疫苗可以有效地激活Th1细胞免疫应答，分泌高水平的细胞因子IFN-γ；可诱导出较普通DC疫苗更明显的抗骨髓瘤作用，并具有一定的安全性。     

D183.49壳聚糖对脐血造血干细胞移植小鼠免疫重建的研究

目的：探讨D183．49壳聚糖对脐血造血干细胞的体外扩增作用及其对脐血造血干细胞移植小鼠的免疫重建并初步探索对脐血造血干细胞移植有调控作用的药物筛选方法。方法：分离脐血中单个核细胞体外扩增培养，测定细胞增殖率及CD34^＋细胞数。60Coγ射线亚致死量照射小鼠，尾静脉注射扩增培养后的单核细胞，每日腹腔注射D183．49壳聚糖，28天。观察记录生存状况；测定血象及CD3、CD4、CD8、CD19水平；脾脏病理切片观察。结果：①体外研究：D183．49壳聚糖10μg组体外增殖效率最显著，48小时达峰值；流式细胞仪测定CD34^＋细胞扩增8倍，P〈0．01。②体内研究：移植药物小鼠血象恢复明显短于其他实验组；CD3、CD4、CD8、CD19水平基本恢复正常，P〈0．05。小鼠脾脏病理切片显示亦恢复正常。结论：D183．49壳聚糖对脐血造血干细胞有明显扩增作用，其能够促进脐血造血干细胞移植小鼠免疫造血重建。     

卡介苗作为抗肿瘤重组蛋白疫苗佐剂的研究

目的：探讨卡介苗（BCG）能否增强抗肿瘤疫苗HSP-MUC1的特异性抑瘤作用，从而将BCG开发为肿瘤疫苗的新型佐剂。方法：动物水平上，BCG和HSP-MUC1共免疫小鼠，观察BCG能否增强HSP-MUC1所激发的特异性抑瘤作用。细胞水平上对BCG发挥佐剂作用的机制进行探讨，将BCG和HSP-MUC1共刺激树突状细胞（DC），观察BCG能否协同HSP-MUC1刺激DC表面的CD86分子表达的上调；并对DC培养上清中IL-6、TNF-α的水平进行测定。结果：BCG＋HSP-MUC1组小鼠的肿瘤重量显著地低于HSP-MUC1组（P〈0.05）。细胞学试验显示，BCG能够显著增强HSP-MUC1对DC的激活作用，使DC表面的CD86分子显著上调。BCG＋HSP-MUC1组的DC培养上清中细胞因子IL-6、TNF-α的水平显著地高于HSP-MUC1组（P〈0.05）。结论：BCG能够显著地增强HSP-MUC1的抗肿瘤活性，具有肿瘤疫苗佐剂的良好功效。     

慢性乙型肝炎患者抗病毒治疗前后外周血树突状细胞和淋巴细胞亚群的变化及意义

目的 分析慢性乙型肝炎患者拉米夫定抗病毒治疗前后外周血树突状细胞和淋巴细胞亚群变化。方法 对16例拉米夫定抗病毒治疗有效的慢性乙型肝炎患者动态观察48周，在治疗前、后对外周血单核细胞来源的树突状细胞进行体外培养，用流式细胞仪检测DC表面分子及淋巴细胞亚群水平，观察治疗前、后的变化。结果 16例患者中，11例治疗持续有效，未发生YMDD变异，5例发生变异。无变异组的慢性乙肝患者，治疗12周时，HLA—DR水平较治疗前降低（P〈0．05）；治疗48周时，CD80、CD40和CD1a与治疗前比明显提高（P〈0．05）而HLA—DR恢复至治疗前水平。变异组患者。治疗12周时，CD83及HLA—DR降低（P〈0．05）；治疗48周时，HLA-DR仍低于治疗前（P〈0．05）。无变异组患者，治疗12周时，CD4^＋T细胞、CD8^＋T细胞、NK细胞、CD19^＋B细胞和CD4^＋／CD8^＋T细胞比值均无明显变化；48周时，CD4^＋T细胞比例增高，NK细胞比例下降（P〈0．05）。而变异组患者的淋巴细胞亚群则无显著改变。结论 拉米夫定治疗持续有效组，DC表面共刺激分子CD80、CD40可随着HBV被长时间有效抑制而得到部分恢复，治疗过程中有DC表面的HLA-DR暂时降低，而后HLA—DR水平恢复和CD1a明显提高，同时外周血CD4^＋T细胞比例上升和NK细胞比例下降。而发生YMDD变异组。DC表面HLA—DR呈现持续性降低。     

转染肿瘤总RNA的DC疫苗诱导抗肝癌特异性免疫

目的：探讨转染人肝癌总RNA的树突状细胞(DC) 疫苗体外诱导特异性细胞毒性T淋巴细胞(CTL)的作用。 方法： 采用原发性肝癌（HCC）病人外周血单核细胞（PBMC），在粒/巨细胞集落刺激因子(GM-CSF)和白细胞介素-4(IL-4) 刺激下增殖分化为DC细胞；从人肝癌细胞中体外扩增肝癌RNA。以HCCRNA转染DC细胞，并与PBMC混合培养诱导扩增CTL。MTT法测定CTL的杀瘤活性。 结果： 转染HCCRNA 48 h后， DC表面分子CD83、CD86和HLA-DR表达明显增高。转染HepG-2细胞HCCRNA的DC和病人HCCRNA诱导的CTL对HepG-2细胞和病人HCC细胞的杀瘤活性均明显高于正常肝细胞RNA+DC、脂质体+DC、Opti-MEM+DC以及空白对照组；而对胃癌SGC-7901细胞无杀伤活性。 结论： 以肝癌RNA为肿瘤抗原，DC作为疫苗的抗原提呈细胞，体外冲击致敏DCs，能诱导肝癌特异性CTL。本研究为HCC术后复发和转移的防治提供一种可能有效的疫苗治疗方法。     

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.     

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.     

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.     

'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.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

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.