IL-17A促进中耳上皮细胞凋亡加重组织损伤

目的建立肺炎链球菌性急性中耳炎小鼠模型,研究白细胞介素17A(interleukin 17A,IL-17A)促进中耳上皮细胞凋亡以及凋亡促进中耳组织损伤的作用。方法鼓膜穿刺术建立C57BL/6小鼠和IL-17A缺陷鼠急性中耳炎模型。收集中耳灌洗液,ELISA检测灌洗液中炎症因子和损伤标志物的表达。中耳组织石蜡切片经TUNEL染色和HE染色,观察小鼠中耳上皮细胞的凋亡和中耳组织损伤程度。采用广谱的凋亡抑制剂Z-VAD-FNK抑制凋亡后,观察中耳组织损伤的变化。结果 IL-17A显著上调白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)和髓过氧化物酶(MPO)表达,加重了中耳炎症。IL-17A还促进了上皮细胞的凋亡和黏膜损伤。抑制凋亡后,显著降低了IL-17A介导的炎症反应,减轻了中耳组织损伤。结论 IL-17A可促进上皮细胞凋亡,加重中耳组织损伤。     

融合蛋白DnaJ-△A146Ply对小鼠肺炎链球菌脑膜炎感染的保护效果

目的 探究融合蛋白DnaJ-ΔA146Ply对小鼠肺炎链球菌脑膜炎感染的保护效果。方法 侧脑室法构建模型的免疫保护实验:C57BL/6雌鼠随机分为抗DnaJ-ΔA146Ply血清+Ⅲ型组、WT血清+Ⅲ型组、抗DnaJ-ΔA146Ply血清+Tigr4组、WT血清+Tigr4组,经侧脑室注射血清及肺炎链球菌血清型Ⅲ型和Tigr4,构建被动免疫模型,24 h后计数菌载量、ELISA测定脑匀浆炎症因子情况及脑组织切片HE染色,观察抗DnaJ-ΔA146Ply血清被动免疫保护效果。尾静脉法构建模型的免疫保护实验:C57BL/6雌鼠随机分为DnaJ-ΔA146Ply组和PBS组,经3次皮下免疫后,通过尾静脉注射肺炎链球菌血清型Tigr4构建肺炎链球菌脑膜炎,24 h后进行脑匀浆、心脏血等菌载量计数,观察免疫保护作用。结果 侧脑室法注射抗DnaJ-ΔA146Ply血清并进行肺炎链球菌攻毒后,脑组织中细菌载量明显降低,脑中TNF-α表达量下调,侧脑室及软脑膜炎症细胞浸润情况减轻。小鼠3次皮下免疫DnaJ-ΔA146Ply后抗体效价达到106,在尾静脉法构建的Tigr4脑膜炎小鼠中,免疫组脑组织中细菌载量降低。结论 融合蛋白DnaJ-ΔA146Ply对不同血清型肺炎链球菌导致的小鼠脑膜炎感染具有保护作用,能够降低细菌侵袭和脑组织炎性反应。     

血管紧张素Ⅱ在EAM小鼠中的表达及致病作用的初步研究

目的探讨EAM小鼠血管紧张素Ⅱ(ATⅡ)水平及其在EAM发生中的作用。方法首先诱导EAM模型,ELISA检测ATⅡ的表达水平,然后实验分为对照组、EAM组、EAM+氯沙坦(ATⅡ抑制剂)组,第21天处死小鼠,HE染色观察心肌炎症浸润情况;RT-q PCR检测心脏组织中相关炎症因子(IL-1β、IL-6、TGF-β)的m RNA的表达水平;Transwell实验检测ATⅡ对巨噬细胞迁移作用及氯沙坦的抑制作用。结果成功诱导小鼠心肌炎模型,心肌组织中有大量的淋巴细胞浸润,ATⅡ水平增高,具有促进炎症因子释放及巨噬细胞迁移的作用;使用ATⅡ抑制剂氯沙坦治疗后,心肌组织炎症评分降低,减少了淋巴细胞浸润;IL-1β、IL-6水平降低,而抑炎因子TGF-β表达高于EAM组;ATⅡ对巨噬细胞的迁移作用受到抑制。结论 ATⅡ可能是EAM发生的重要因素,抑制ATⅡ能够有效缓解心肌炎症损伤。     

NKT细胞在MRSA感染性肺炎中作用的初步研究北大核心CSCD

目的:初步探讨NKT细胞在耐甲氧西林金葡菌(MRSA)感染性肺炎模型中的作用。方法:MRSA经鼻黏膜感染C57BL/6野生型(WT)小鼠和CD1d^(-/-)小鼠。观察小鼠生存率,检测肺组织中细菌载量,ELISA检测肺组织中细胞因子TNF-α、IFN-γ、IL-17、IL-6含量,流式细胞术检测肺组织中CD3^(+)CD8^(+)T细胞、NK细胞和NKT细胞百分比。结果:MRSA致死剂量感染5 d,WT小鼠生存率显著低于CD1d^(-/-)小鼠(46.2%vs 90.0%,P<0.05)。感染12~48 h,WT小鼠肺组织中细菌载量显著高于CD1d^(-/-)小鼠(P<0.01);WT小鼠肺组织中TNF-α、IFN-γ、IL-6和IL-17含量较CD1d^(-/-)小鼠明显增高(P<0.01)。MRSA感染24 h,WT小鼠肺组织中NKT细胞、NK细胞百分比显著高于CD1d^(-/-)小鼠(P<0.01),而CD3^(+)CD8^(+)T细胞百分比低于CD1d^(-/-)小鼠(P<0.01)。结论:在MRSA肺炎中,NKT细胞被激活,从而加重肺部炎症反应,降低生存率,减缓细菌清除。     

CD38蛋白下调炎症因子的表达减轻脂多糖联合D-氨基半乳糖诱导的急性肝脏损伤

目的研究CD38蛋白在脂多糖联合D-氨基半乳糖(LPS/D-GalN)所诱发的小鼠急性肝损伤中的作用。方法野生型C57BL/6小鼠(WT)和CD38基因敲除小鼠(CD38 KO)随机分为正常对照组,模型早期组和模型晚期组。在造模后2 h和6 h处死动物,检测血清谷丙转氨酶(ALT)和谷草转氨酶(AST),实时荧光定量PCR检测肝脏组织炎症因子的表达,HE染色检测组织病理改变。结果在LPS/D-GalN诱导的模型小鼠中,与WT小鼠相比,CD38 KO小鼠血清ALT和AST水平显著升高,肝脏组织中炎性因子白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)以及γ干扰素(IFN-γ)的表达水平显著升高;病理组织学检测显示肝脏组织出血严重,肝实质细胞空泡变形明显增多,肝细胞死亡显著增加。结论 CD38蛋白通过下调炎症因子的表达,减少肝细胞死亡,减轻LPS/D-GalN诱导的急性肝脏损伤。     

幼年期肺炎链球菌肺炎对哮喘形成的影响

目的研究幼年期肺炎链球菌肺炎(S.pneumoniaepneumonia,S.pp)对成年期哮喘形成的影响。方法 3周龄雌性Balb/c小鼠随机分为对照组(control)、幼年期肺炎链球菌肺炎组(Inf S.pp)、哮喘组(OVA)、幼年期肺炎链球菌肺炎哮喘组(Inf S.pp+OVA)。3周龄雌性Balb/c小鼠鼻腔滴注2×106cfu/ml肺炎链球菌25μl建立肺炎链球菌肺炎模型,成年期用OVA致敏激发Balb/c小鼠建立哮喘模型,最后1次激发完成24 h内检测小鼠气道高反应性,HE染色制作肺组织病理切片,观察组织病理变化,检测支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中细胞总数、细胞分类计数及细胞因子水平,流式细胞术检测小鼠肺组织中CD4~+T细胞水平。结果幼年期肺炎链球菌肺炎哮喘组肺组织炎症细胞侵润、气道反应性、BALF细胞总数、嗜酸性粒细胞和中性粒细胞及细胞因子IL-5、IL-13、IL-17水平显著高于对照组(P0.05),IFN-γ、IL-10表达显著降低(P0.05),肺组织内Th2、Th17水平显著高于对照组(P0.05),Foxp3~+Treg水平显著降低(P0.01),Th1/Th2、Foxp3~+Treg/Th17比例明显降低(P0.01)。幼年期肺炎链球菌肺炎哮喘组肺组织炎症细胞侵润、气道高反应性、BALF细胞分类计数及炎性因子水平、肺组织CD4~+T细胞水平与哮喘组比较无统计学差异。结论幼年期肺炎链球菌肺炎对成年期哮喘形成无显著影响。     

小鼠克雷白杆菌肺炎模型的建立及检测

目的:研究有效建立小鼠克雷白杆菌肺炎模型的方法并进行相关炎症因子检测。方法:构建动物模型,C57小鼠随机分为对照组和模型组,应用气管内注射肺炎克雷白杆菌(106cfu,20#l)的方法建立肺炎模型,感染后24 h,观察小鼠一般状态;收集肺泡灌洗液、分离肺组织,大体观察;肺组织切片HE染色观察两组小鼠炎症情况;肺泡灌洗液进行中性粒细胞计数,肺泡灌洗液和肺组织MPO检测的方法来比较两组小鼠中性粒细胞浸润情况;应用实时定量PCR、Western blot对两组小鼠相关炎症因子进行检测。结果:和对照组相比,模型组小鼠出现明显喘息,寒颤以及竖毛;肺部出现明显的充血、水肿;HE染色显示模型组肺泡壁增厚,大量炎症细胞浸润;模型组肺泡灌洗液中性粒细胞的数量明显高于对照组(P<0.05);模型组支气管肺泡灌洗液和肺组织中MPO表达水平均明显高于对照组(P<0.05);实时定量PCR、Western blot结果均表明模型组小鼠肺组织中炎症因子较对照组明显升高。结论:利用气管内注菌建立小鼠克雷白肺炎模型的方法确实可靠,该模型可用于细胞因子及其受体与克雷白杆菌肺炎关系的研究。     

Lgals3在分枝杆菌感染与肉芽肿形成过程中的作用及机制研究北大核心CSCD

目的:研究宿主Lgals3分子在分枝杆菌感染与肉芽肿形成过程中的作用及机制。方法:检测Lgals3在H37Rv气溶胶感染的小鼠、家兔肺组织及临床结核病(TB)患者样本中的表达与分布;运用CRISPR/Cas9技术构建lgals3基因全身敲除(lgals3^(−/−))小鼠,并建立lgals3^(−/−)小鼠海分枝杆菌(Mm)尾静脉感染模型,观察小鼠尾组织损伤与脓肿形成,比较尾部大体病变;HE染色观察小鼠尾组织病理学变化及肉芽肿形成差异;抗酸染色观察细菌分布;小鼠尾组织匀浆铺板接种于7H10-OADC培养基后比较菌载量;免疫组化和图像分析技术观察比较小鼠尾组织中Lgals3和TGF-β、MMP-9的表达与分布。结果:H37Rv气溶胶感染的小鼠、兔子肺组织及临床结核患者肉芽肿区域中Lgals3呈强阳性表达。应用CRISPR/Cas9技术成功获得lgals3^(−/−)小鼠;Mm感染后,lgals3^(+/+)小鼠尾组织出现明显的渐进性脓肿及溃疡,而lgals3^(−/−)小鼠仅出现散在的小面积脓肿,且差异具有统计学意义(P<0.01)。HE染色观察发现lgals3^(+/+)小鼠感染Mm后伴随明显炎症细胞浸润,且形成典型肉芽肿样病变,而lgals3^(−/−)小鼠仅有少量炎症细胞分布。组织匀浆铺板计数发现,相较于lgals3^(+/+)小鼠,lgals3^(−/−)小鼠菌载量显著降低(P<0.01)。免疫组化和图像分析技术发现,lgals3^(+/+)小鼠感染Mm后Lgals3在肉芽肿区域内呈弥散分布,TGF-β和MMP-9表达上调并呈强阳性表达;lgals3^(−/−)小鼠仅检测到少量TGF-β和MMP-9。结论:Mm感染可诱导宿主Lgals3的表达,并在TGF-β和MMP-9协同作用下促进肉芽肿形成,帮助细菌逃避早期宿主免疫应答。     

CD226基因敲除加重小鼠放射性肝损伤的发生

目的观察CD226基因敲除(KO)对小鼠放射性肝损伤的影响。方法雄性野生型(WT)和CD226KO小鼠随机分为非辐照组和辐照组,非辐照组不经过任何处理,辐照组接受8 Gy ~(60)Co照射建立急性放射性损伤模型。观察小鼠存活和体质量改变情况, HE染色观察肝脏和结肠病变情况,实时定量PCR检测肝和结肠组织炎症因子诱导型一氧化氮合酶(iNOS)、白细胞介素1β(IL-1β)、 IL-6、 IL-12p40、肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)的mRNA水平。结果与辐照WT小鼠相比,辐照后的CD226KO小鼠体质量明显下降,存活率下降; CD226KO小鼠肝脏损伤加重,肝指数(肝质量/体质量)下降显著, iNOS、 IL-1β、 IL-6、 IL-12p40、 TNF-α、 MCP-1 mRNA水平显著升高;但CD226KO小鼠结肠炎症减轻,炎症因子mRNA水平降低。结论 CD226缺失降低辐照小鼠存活率,与肝损伤加重密切相关。     

miR-221在哮喘患儿痰液的表达及对哮喘小鼠气道炎症的影响

目的探讨miR-221在哮喘患儿及哮喘小鼠中的表达及其对气道炎症的作用。方法应用realtime PCR方法检测哮喘患儿和健康儿童痰液,以及卵清蛋白诱导的哮喘小鼠和正常小鼠肺组织中miR-221的表达。化学合成miR-221抑制剂与阴性对照,并分别鼻滴至哮喘小鼠,HE染色验证哮喘小鼠模型的成功建立,免疫荧光方法和western blot方法检测各组小鼠肺组织IL-1β与IL-4的蛋白表达。结果 HE染色显示哮喘模型组小鼠炎症细胞浸润明显高于正常对照组,哮喘小鼠模型成功建立。与对照组相比,哮喘患儿痰液及哮喘小鼠肺组织中miR-221表达水平显著升高(P0.01)。哮喘小鼠肺组织IL-1β与IL-4的蛋白表达水平显著高于正常对照组,而低于miR-221抑制剂组(P0.01)。结论 miR-221在支气管哮喘患儿及支气管哮喘小鼠模型中高表达,miR-221抑制剂能够抑制哮喘小鼠的气道炎症。     

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.     

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.     

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.     

'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.     

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 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.