表达ESAT-6-gpi和IL-21的B16F10瘤苗的构建及其活性鉴定

为构建膜表达糖基化磷脂酰肌醇(GPI)锚定的结核杆菌早期分泌靶抗原6 kD(ESAT-6)和分泌IL-21的B16F10瘤苗并鉴定其活性,利用重叠PCR法构建pIRES-ESAT-6-gpi/IL-21重组质粒,以脂质体转染重组质粒到B16F10细胞,G418筛选出阳性克隆,用RT-PCR、免疫荧光、FCM和Western blot检测瘤苗细胞靶抗原表达,用瘤苗细胞培养上清刺激小鼠CD8+T细胞,检测瘤苗所分泌IL-21的生物学活性。结果表明,pIRES-ESAT-6-gpi/IL-21重组质粒DNA测序正确,B16F10-ESAT-6-gpi/IL-21瘤苗细胞目的基因ESAT-6表达于瘤苗细胞表面,增殖能力未受外源基因导入影响,分泌的IL-21具有生物学活性,为研究膜表达ESAT-6和分泌表达IL-21瘤苗的抗瘤效应奠定了基础。     

糖基化磷脂酰肌醇修饰的小鼠IL-21瘤苗抗肿瘤效应及其机制初探

目的 构建糖基化磷脂酰肌醇(glycosyl phosphafidylinositol,GPI)修饰的小鼠IL-21瘤苗,并对此瘤苗的抗肿瘤效应及其机制作初步探讨.方法 通过重叠PCR方法获得IL-21-GPI融合基因并将其插入空载体pcDNA3.1.将鉴定过的重组载体以脂质体法转染B16F10细胞制成瘤苗,细胞间接免疫荧光法及流式细胞仪检测转染瘤细胞膜表面IL-21的表达,通过对小鼠脾细胞的增殖作用鉴定表达的IL-21的生物学活性.将瘤苗接种小鼠后,通过观察小鼠肿瘤体积和生存率分析瘤苗的抗瘤性,并检测了瘤苗免疫鼠的细胞免疫活性.结果 正确构建了pcDNA3.1/IL-21-GPI重组载体,膜表达的IL-21有良好的生物学活性,制备的瘤苗能发挥抗肿瘤效应,其机制与免疫鼠细胞免疫活性增强有关.结论 成功构建了具有抗肿瘤活性的GPI修饰的IL-21瘤苗,为其进一步抗肿瘤免疫治疗研究奠定了基础.     

B16F10/ESAT-6-GPI-IL-21瘤苗安全性及抗肿瘤效应研究

目的:评估B16F10/ESAT-6-GPI-IL-21瘤苗用于C57BL/6小鼠的安全性及其诱导的抗肿瘤免疫效应。方法:应用免疫荧光、FCM检测瘤苗细胞ESAT-6-GPI的表达情况;以Western blot方法检测瘤苗细胞IL-21的表达情况;动物实验检测瘤苗体内应用的安全性;制备荷瘤鼠术后免疫模型,评价瘤苗免疫效果。结果:B16F10/ESAT-6-GPI-IL-21瘤苗细胞表面有靶抗原ESAT-6-GPI表达,并分泌IL-21。于C57BL/6小鼠皮下接种2×105个B16F10/ESAT-6-GPI-IL-21瘤苗细胞,60天内未见致瘤,但能够诱导小鼠产生有效的抗肿瘤免疫效应。结论:B16F10/ESAT-6-GPI-IL-21瘤苗致瘤性明显下降,低剂量应用具有安全性,能够诱导小鼠产生有效的抗肿瘤免疫效应。     

GPI修饰的ESAT-6核酸疫苗的构建及其免疫功能初探

目的构建糖基化磷脂酰肌醇(GPI)修饰的结核杆菌早期分泌性抗原靶(ESAT-6)核酸疫苗,初步分析其免疫功能。方法利用重叠PCR法构建pIRES-ESAT-6-gpi重组体,转染B16F10细胞,G418筛选阳性克隆,用RT-PCR、免疫荧光检测转染细胞的ESAT-6抗原表达情况;采集ESAT-6核酸疫苗免疫鼠血清,分别检测抗体滴度和CDC效应,免疫磁珠法分选CD4+和CD8+T细胞,CFSE/7-AAD细胞毒实验分析CTL细胞毒活性。结果测序正确的重组体pIRES-ESAT-6-gpi转染细胞后,ESAT-6表达于细胞膜表面。ESAT-6核酸疫苗免疫血清和CD8+T细胞分别通过CDC效应和细胞毒作用杀伤膜表达ESAT-6的B16F10细胞。结论构建的GPI修饰的ESAT-6核酸疫苗能够诱导免疫鼠产生体液和细胞免疫反应,杀伤膜表达ESAT-6的B16F10细胞,为进一步基于该法构建B16F10瘤苗的抗肿瘤免疫效应及机制研究奠定了基础。     

小鼠白细胞介素21 cDNA的克隆及真核表达质粒的构建

目的：克隆小鼠自细胞介素2l(IL-21)基因，构建真核表达质粒，用以进行肿瘤的基因治疗。方法：用RT-PCR法。从ConA活化的小鼠T细胞中扩增IL-21 cDNA。克隆人哺乳动物细胞高效表达质粒pcDNA3．1中，构建重组mIL-21真核表达质粒。重组体用载体上的通用引物和PCR下游引物为测序引物，鉴定克隆的正确性。将已鉴定的重组质粒用脂质体法转染Sp2／0细胞，用RT-PCR法鉴定转染细胞中IL-21基因的表达，用MTT比色法检测表达的mIL-21诱导的NK细胞杀伤活性的增强。结果：正确构建了重组真核表达质粒pcDNA3．1／mIL-21，并在转染的细胞中检测出IL-2l的表达，表达的mIL-21可在体外增强NK细胞的杀伤活性：结论：成功地构建了重组真核表达质粒pcDNA3．1／mIL-21，为进一步在肿瘤动物模型中进行IL-21基因治疗及疗效观察奠定了基础.     

IL—6基因转染的瘤苗体内诱导的抗肿瘤免疫功能与效果

经丝裂霉素C体外处理后,将IL-6基因转染的、高分泌的IL-6的B16黑色素瘤细胞制成瘤苗。结果发现,体内注射IL-6基因转染的瘤苗后,小鼠脾脏CTL活性、NK活性及IL-2诱导的LAK活性显著升高。经IL-6基因转染瘤苗体内治疗后,荷瘤小鼠的皮下肿瘤生长显著减慢、肺转移结节数显著降低、存活期显著延长,若同时合用低剂量IL-2,则上述治疗效果更好。可见IL-6基因转染的瘤苗能有效地通过诱导机体抗肿瘤免疫功能而发挥抗肿瘤作用,与低剂量IL-2合用后,IL-6基因转染的瘤苗的抗肿瘤效果更佳。     

GM-CSF与IL-21基因共转染卵巢癌细胞及其对NK细胞活性的影响

 目的 探讨粒细胞巨噬细胞集落刺激因子（GM-CSF）与 IL-21 基因共转染的体外抗肿瘤效应。 方法 制备 pRSC-GM-CSF- IL21 重组质粒，再用脂质体将 质粒 pRSC-GM-CSF、pRSC-IL21 和 pRSC-GM-CSF-IL21 分别转染人卵巢癌 SKOV3 细胞（依次命名为 SKOV3/GM- CSF、SKOV3/IL21、SKOV3/GM-CSF-IL21 细胞）。以 RT- PCR 方法检测 GM-CSF 与 IL-21 表达；倒置相差显微镜观察各组细胞形态学变化；噻唑蓝法测定各组细胞增殖活性；流式细胞仪检测各组细胞细胞周期分布，细胞表面 HLA-ABC、主要组织相容性复合体 I 类相关基因（MIC A/B）、细胞间黏附分子 1（ICAM-1）的表达，以及各组细胞培养上清液对 NK 细胞活性的影响。以未转染的 SKOV3 细胞和转染空质粒 pRSC 的细胞（SKOV3/Neo）为对照。 结果 经酶切与测序鉴定，pRSC-GM-CSF-IL21 重组体构建正确，共转染 SKOV3 细胞中有目的基因 GM-CSF、IL-21 的表达。各组细胞的形态学、增殖特性、细胞周期分布及 HLA-ABC 表达无明显变化。SKOV3、SKOV3/Neo、SKOV3/GM-CSF、SKOV3/IL21 和 SKOV3/GM-CSF-IL21 细胞的 MIC A/B 表达量分别为 19.24% ± 2.31%、31.11% ± 3.76%、37.27% ± 3.04%、54.97% ± 4.77% 和 63.94% ± 5.90%；ICAM-1 表达量分别为 35.88% ± 3.06%、37.75% ± 4.09%、68.59% ± 4.84%、78.53% ± 5.78% 和 84.10% ± 3.98%；加入各组细胞培养上清液后 NK 细胞活性分别为 31.8% ± 3.6%、42.7% ± 3.3%、54.7% ± 7.4%、55.9% ± 4.4% 及 63.4% ± 6.4%。SKOV3/GM-CSF-IL21 细胞分别与 SKOV3、SKOV3/ Neo 细胞比较，3 项指标的差异均有统计学意义（P < 0.01）。 结论 GM-CSF 与 IL-21 基因共转染可上调 SKOV3 细胞表面 MIC A/B 与 ICAM-1 表达，表达产物可增强 NK 细胞活性，这有助于免疫细胞激活，增强抗肿瘤效应。     

新型细胞因子IL-24真核表达载体的构建、表达及其对肿瘤的抑制作用

目的：构建IL-24真核表达质粒，并研究其体内外表达对肿瘤细胞的生长抑制作用。方法：采用重组DNA技术构建IL-24真核表达质粒pEGFP-C1-IL-24。用脂质体法将重组质粒及空载体外转染HIC细胞，再经激光扫描共聚焦显微镜（LSCM）观察其表达，用MTT法检测HIC细胞的体外增殖能力，用流式细胞仪检测细胞周期与凋亡。小鼠皮下接种转染IL-24真核表达质粒或空载的B16细胞观察其体内致瘤性的变化。小鼠实体瘤模型研究基因转染对肿瘤的生长抑制作用。结果：pEGF-C1-IL-24转染HIC细胞后，LSCM可观察到其表达。IL-24基因转染的HIC细胞体外增殖能力明显受到抑制．G2-M期细胞比例增高，细胞被阻滞在G2-M期。转染IL-24的B16细胞体外致瘤率降低。与对照组相比．IL-24基因治疗组肿瘤生长明显受到抑制（P〈0.05）。结论：IL-24基因转染的肿瘤细胞体外生长受抑。瘤内注射pEGFP-C1-IL-24可抑制肿瘤生长，具有明显的抗肿瘤作用。     

IL-21修饰的脐血间充质干细胞对卵巢癌裸鼠治疗作用

探讨IL-21修饰的脐血间充质干细胞(UMSC)对荷卵巢癌裸鼠的治疗作用。从脐血单个核细胞中培养UMSC,经流式细胞术(FCM)鉴定后用于重组体pIRES2-IL-21-EGFP转染。RT-PCR检测UMSC中IL-21的表达。荷瘤裸鼠治疗实验分为PBS、UMSC、UMSC空质粒和UMSC-IL-21四组。以肿瘤大小、荷瘤鼠生存期判断IL-21修饰的UMSC对荷瘤鼠的治疗效应。以ELISpot法检测脾细胞IFN-γ分泌、免疫荧光法检测肿瘤组织中IL-21、NKG2D的表达,并同时检测了NK细胞活性。结果显示,脐血分离的单个核细胞经三代培养后已初步符合UMSC表型,pIRES2-IL-21-EGFP转染的UMSC能表达IL-21和EGFP。UMSC-IL-21治疗组,能抑制肿瘤生长,延长荷瘤裸鼠生存期,与其他组相比,差异有明显统计学意义(P0.01)。UMSC-IL-21组肿瘤局部能表达IL-21与NKG2D、脾细胞分泌IFN-γ能力升高,NK细胞杀伤活性增强,表明UMSC-IL-21有良好地抗裸鼠卵巢癌作用,其机制可能与表达IL-21诱导IFN-γ分泌增加,激活NK细胞活性有关。     

人IL-24基因的克隆及在COS-7细胞中的表达

目的：克隆人IL-24基因，构建真核表达载体，在COS-7细胞中进行表达，并检测重组表达蛋白hIL-24的抗肿瘤活性。方法：分离人外周血单个核细胞，提取细胞总RNA，采用RT-PCR技术克隆人IL-24基因，将其重组于pUC19，双脱氧末端终止法测定其核苷酸序列，构建真核重组表达载体pcDNA3-hIL-24，进行双酶切和PCR鉴定，以质粒pcDNA3-hIL-24转染COS-7细胞进行瞬时表达，用RT-PCR检测mRNA表达，并用MTT法、TUNEL法和流式细胞术检测其诱导肿瘤细胞凋亡的活性。结果：成功获得621bp的人IL-24基因，测序正确，经双酶切和PCR鉴定真核表达质粒构建正确，以脂质体转染COS-7细胞后，用RT-PCR法、MTT法、TUNEL法和流式细胞术检测表明COS-7细胞可表达hIL-24，且所表达的人IL-24具有较强的诱导A549肺腺癌细胞凋亡的作用。结论：人IL-24基因的瞬时表达和凋亡效应的初步研究已获成功，为研究人IL-24抗肿瘤的分子机制和临床应用提供了实验依据．     

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.     

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.     

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.     

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.     

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.     

Environmental risk factors and their role in the management of atopic dermatitis

Introduction: The etiology of atopic dermatitis (AD) is multifactorial with interaction between genetics, immune and environmental factors.

Areas covered: We review the role of prenatal exposures, irritants and pruritogens, pathogens, climate factors, including temperature, humidity, ultraviolet radiation, outdoor and indoor air pollutants, tobacco smoke exposure, water hardness, urban vs. rural living, diet, breastfeeding, probiotics and prebiotics on AD.

Expert commentary: The increased global prevalence of AD cannot be attributed to genetics alone, suggesting that evolving environmental exposures may trigger and/or flare disease in predisposed individuals. There is a complex interplay between different environmental factors, including individual use of personal care products and exposure to climate, pollution, food and other exogenous factors. Understanding these complex risk factors is crucial to developing targeted interventions to prevent the disease in millions. Moreover, patients require counseling on optimal regimens for minimization of exposure to irritants and pruritogens and other harmful exposures.     

Class II HLA in Georgia Caucasus Tbilisi Georgians and their Mediterranean ancestry: The Usko Mediterranean languages

Georgia (or Sakartvelo in its own language) is a South Caucasus Mts. country with its easternmost part is enigmatically named Iberia, like the Iberian Peninsula, which may refer to rivers “Kura” and “Ebro” or their valleys respectively. Most of their inhabitants speak Georgian which is included within Dene-Caucasian group and Usko-Mediterranean subgroup of languages. The latter includes Basque, Berber, ancient Iberian-Tartessian, Etruscan, Hittite, Minoan Lineal A and others. In the present paper, HLA class II -DRB1 and -DQB1 alleles has been studied and extended haplotypes calculated. Most frequent haplotypes are also of Mediterranean origin (i. e.: (A*02-B*51)-DRB1*11:01-DQB1*03:01, (A*02-B*51)-DRB1*13:01-DQB1*06:03, or (A*24-B*35)-DRB1*01:01-DQB1*05:01) and DA genetic distances show that closest world populations to Georgians are Mediterraneans. Georgians also show common extended haplotypes ((A*02-B*51)-DRB1*11:01-DQB1*03:01, (A*02-B*13)-DRB1*07:01-DQB1*02:01 and (A*03-B*35)-DRB1*11:01-DQB1*03:01) with Svan people, a secluded population in North Georgia mountains. We can conclude that Georgians belong to a very old Mediterranean substratum according to both linguistics (Usko Mediterranean languages) and HLA genetics.     

'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.