沙眼衣原体重组口服活疫苗的体液免疫学特性

目的检测所构建的沙眼衣原体(Ct)重组疫苗株口服免疫小鼠后,产生的体液免疫应答。方法将所构建的重组菌以5×108菌落形成单位(CFU)/只的剂量,口服免疫雌性Balb/c小鼠,随机分为7组,每组3只。于免疫后第2,7,14,21,28,35和42d各取1组,采集血清、小肠肠腔冲洗液和阴道冲洗液标本。分别以D型Ct和鼠伤寒杆菌临床分离株为抗原,检测各组血清、小肠冲洗液和阴道冲洗液标本中特异性IgG,IgA和IgM。取抗D型Ct血清IgG阳性标本和阴道冲洗液IgA阳性标本,分别用于检测与C型Ct的交叉反应。结果在血清标本中,有抗D型Ct和抗鼠伤寒杆菌特异性IgG和IgM,在小肠冲洗液和阴道冲洗液标本中有抗上述两种抗原特异性IgG和IgA。抗D型CtIgG阳性血清标本和IgA阳性阴道冲洗液标本与C型Ct均有交叉反应。结论所构建的疫苗株免疫小鼠后,能够诱导Ct特异性IgG及IgA产生。     

蛋白质组学筛选沙眼衣原体感染依赖性免疫优势抗原

目的：采用蛋白质组学方法筛选沙眼衣原体（Chlamydia trachomatis,Ct）感染依赖性免疫优势抗原。方法：将纯化的Ct D型标准株EB经紫外线照射制备UV-EB,然后将未经处理的活EB和紫外线处理的UV-EB分别感染体外培养的HeLa细胞,间接免疫荧光法检测紫外线对EB的灭活效果;分别将低剂量活EB滴鼻感染小鼠或将较高剂量UV-EB经肌肉接种免疫小鼠,首先检测两种接种方式下Ct在体内的增殖情况,然后收集两组小鼠的血清,分别与Ct D型全基因组908个开放读码框编码的GST-Ct融合蛋白进行ELISA反应,蛋白质组学筛选仅与活EB感染组小鼠血清反应且反应频率高的Ct感染依赖性免疫优势抗原;最后用筛选到的抗原体外刺激Ct生殖道感染小鼠的脾细胞,ELISA法检测细胞因子IFN-γ的产生情况。结果：经紫外线灭活的UV-EB体外不能感染HeLa细胞,肌肉接种免疫小鼠后也不能在小鼠肌肉组织内增殖,而活EB滴鼻感染组小鼠的肺组织中可检出大量的Ct;活EB滴鼻感染组小鼠与UV-EB肌肉接种免疫组小鼠的血清抗Ct抗体滴度差异无显著统计学意义;将两组小鼠血清分别与Ct全基因组编码的蛋白反应,通过蛋白质组学方法共鉴定出60个抗原能被至少1份小鼠血清识别;22个抗原能同时被两组或任一组血清以≥50%的频率识别,为Ct免疫优势抗原,其中5个抗原仅被活EB滴鼻免疫组血清优势识别,即为Ct感染依赖性免疫优势抗原;该类抗原体外刺激小鼠脾细胞,能诱导良好的Th1型细胞免疫回忆应答。结论：筛选获得了Ct D型感染依赖性免疫优势抗原,为Ct亚单位疫苗研究提供了新的研究靶标。     

E-选择素在生殖道沙眼衣原体感染中的作用研究

目的 研究E-选择素在生殖道沙眼衣原体感染中的作用.方法 将64只小鼠生殖道沙眼衣原体感染模型随机分成2组,实验组中的小鼠用人工合成E-选择素干预,对照组中的小鼠用生理盐水进行干预,统计两组在1w,2w,3w,4w不同时间点上的感染小鼠阴道脱落菌量以及沙眼衣原体感染率.结果 成功建立生殖道沙眼衣原体感染的小鼠模型:与对照组相比较,人工合成E-选择素干预后1w和2w的生殖道沙眼衣原体感染率(90.2％vs 77.4％,85.3％vs 70.1％)显著增高,而阴道脱落菌量显著增高(4.758 ±0.225 vs 3.210 ±0.315,2.698±0.177 vs 1.809±0.203),两组间差异具有统计学.在干预后3w两组的沙眼衣原体感染率分别为34％和24％,阴道脱落菌量分别为1.412±0.134和1.201±0.189,两组间差异无统计学意义;两组在干预后4w的沙眼衣原体感染率均为0且无阴道脱落菌量.结论 E-选择素能促进生殖道沙眼衣原体感染的发生率以及增加沙眼衣原体的毒力,本文为进一步阐明生殖道沙眼衣原体感染的发病机制提供实验依据.     

TLR2介导沙眼衣原体感染早期免疫应答的机制研究

目的 探讨TLR2在小鼠沙眼衣原体生殖道早期感染时的免疫应答中的作用.方法 沙眼衣原体小鼠肺炎株(MoPn)经生殖道分别感染野生型小鼠(WT=11只)、TLR2基因缺陷小鼠(TLR2 KO=14只),建立生殖道沙眼衣原体感染模型.分别于感染后不同时间点取阴道棉拭子,获取分泌物,检测分泌物衣原体包涵体数量和炎症细胞因子IL-1α、IL-6和MIP-2水平.结果 TLR2 KO和WT小鼠在每个检测时间点,生殖道分泌物带菌量无差异,且带菌持续时间相同;与WT小鼠比较,在感染后第3d和第10d,TLR2 KO小鼠生殖道分泌物的炎症细胞因子IL-1α、IL-6和MIP-2水平均低于野生型WT小鼠,但差异均无统计学意义(P＞0.05);而在感染后第7d,3种细胞因子均明显低于WT小鼠,且差异有统计学意义(P＜0.05).结论 在沙眼衣原体生殖道感染中,TLR2可能介导了早期炎症细胞因子的产生.     

Toll样受体2(TLR2)和TLR4在沙眼衣原体生殖道感染介导的免疫应答

目的探讨Toll样受体2(TLR2)和TLR4在小鼠沙眼衣原体生殖道感染中的作用。方法用1×10~4包涵体形成单位的Mo Pn沙眼衣原体小鼠肺炎株经生殖道感染野生型(WT)小鼠(n=11)、TLR2基因缺陷(TLR2-/-)小鼠(n=14)和TLR4-/-小鼠(n=11),复制生殖道沙眼衣原体感染模型。在感染后第3、7、10、17、24、31、38、45天取阴道棉拭子,免疫荧光法检测衣原体包涵体数量,ELISA检测白细胞介素1α(IL-1α)、IL-6和巨噬细胞炎性蛋白2(MIP-2)水平。感染后第70天,采用免疫荧光法分析血清抗体类型和效价;用Mo Pn沙眼衣原体感染无菌分离的腹腔巨噬细胞,培养24 h,ELISA测定上清液中IL-1α、IL-6和MIP-2含量;体外用紫外线灭活的衣原体抗原刺激无菌分离的脾细胞,培养72 h,ELISA测定上清液中γ干扰素(IFN-γ)、IL-17、IL-4和IL-5水平。结果与WT小鼠相比,TLR2-/-或TLR4-/-小鼠在各时间点的生殖道带菌量无差异、且带菌持续时间相同,感染后第38天,所有小鼠均清除了下生殖道感染的衣原体。TLR2-/-小鼠巨噬细胞产生的IL-1α、IL-6和MIP-2水平均明显低于WT小鼠,而TLR4-/-小鼠巨噬细胞产生的3种炎症细胞因子水平均与WT小鼠无显著性差异;TLR2-/-小鼠棉拭子标本具有较低水平的炎症细胞因子。所有小鼠脾细胞均产生高水平的IFN-γ和IL-17、较低水平的IL-4和IL-5、血清Ig G2a/Ig G1比值均大于1,但各组间无显著性差异。结论 TLR2而不是TLR4介导了沙眼衣原体生殖道感染的早期免疫应答,但沙眼衣原体诱导的适应性免疫应答可能既不依赖于TLR4,也不依赖于TLR2。     

TLR2信号在沙眼衣原体生殖道感染中介导了早期炎症细胞因子的产生

目的探讨TLR2和TLR4在小鼠沙眼衣原体生殖道感染免疫应答中的作用。方法用沙眼衣原体小鼠肺炎株(Mo Pn,1×104IFUs)经生殖道感染野生型小鼠(WT,11只)、TLR2基因缺陷小鼠(TLR2 KO,14只)和TLR4基因缺陷小鼠(TLR4 KO,11只),复制生殖道沙眼衣原体感染模型。于感染后不同时间点取生殖道分泌物,部分用于免疫荧光法检测衣原体包涵体数量,部分用于炎症细胞因子IL-1α、IL-6和MIP-2水平检测。在感染后第70天,处死小鼠,无菌分离腹腔巨噬细胞,于体外衣原体Mo Pn感染,培养24 h后,测定上清液中IL-1α、IL-6和MIP-2含量。细胞因子含量测定均采用ELISA法。结果 TLR2 KO或TLR4 KO小鼠与WT小鼠在每一个检测时间点下生殖道带菌量无差异,且带菌持续时间相同,截止到感染后第38天,3种基因型所有小鼠均清除了下生殖道感染的衣原体。TLR2基因缺失小鼠巨噬细胞产生的炎症细胞因子IL-1α、IL-6和MIP-2水平均明显低于野生型WT小鼠(P0.01),而TLR4基因缺失小鼠巨噬细胞产生的3种炎症细胞因子水平均与野生型WT小鼠无差异(P0.05);TLR2基因缺失小鼠棉拭子标本同样具有较低水平的炎症细胞因子(P0.05)。结论在沙眼衣原体生殖道感染中,TLR2介导了早期炎症细胞因子的产生。沙眼衣原体诱导的早期免疫应答部分依赖于TLR2,而不依赖TLR4。     

372例不孕症患者支原体、衣原体感染的检测分析

生殖道支原体和衣原体可造成宫颈炎、输卵管的黏连或子宫内膜炎而导致不孕不育,引起流产、早产,是女性生殖道感染的常见病原体,为导致妇女不孕的因素之一[1]。目前临床检测的主要有解脲脲原体(Uu)、人型支原体(Mh)、沙眼衣原体(Ct)等。为了了解不孕症患者Uu、Mh、Ct的感染状况,随机对2010年2月～2011年12月来我院门诊就诊的     

人ACE2转基因小鼠SARS-CoV感染后的病理变化及免疫学反应

目的 观察SARS-Cov感染人血管紧张素转换酶2(hACE2)转基因小鼠引起的病理变化并初步探讨其发生的免疫学机制,为SARS研究提供可靠的动物模型.方法 实时PCR测定hACE2转基因小鼠的拷贝数;用PUMC01株SARS-CoV感染hACE2转基因小鼠,光镜下观察小鼠全身组织器官的病理变化;ELISA方法检测血清特异性抗体和肺组织匀浆上清细胞因子TNF-a、IL-6、IFN-γ变化.结果 单拷贝hACE2转基因小鼠在感染SARS-CoV后肺组织出现更严重的间质性肺炎并伴有肺外多器官损伤;少数转基因小鼠血清检测到特异性IgC,抗体;转基因小鼠肺组织匀浆上清TNF-a、IL-6、IFN-γ的水平明显升高.结论 hACE2转基因小鼠感染SARS-CoV后出现与人类SARS患者相似的病理特征和免疫学反应,为研究SARS发病机制和药物评价提供了小动物模型.     

衣原体噬菌体phiCPG1衣壳蛋白Vp1对沙眼衣原体的作用研究

目的 将原核表达纯化的衣原体噬菌体phiCPG1衣壳蛋白Vp1作用于沙眼衣原体(Chlamydia trachomatis,Ct),以期发现该Vp1蛋白对Ct生长的影响.方法 原核表达并纯化噬菌体phiCPG1的衣壳蛋白Vp1,将纯化后的蛋白复性后与Ct的标准株或临床野生株(终浓度为53μg/ml)室温孵育3h后接种至单层致密McCoy细胞中,培养过程中均加入了了 Vp1蛋白,48 h后碘染色包涵体计数和透射电镜观察Vp1蛋白对Ct生长的影响;MTT法检测Vp1蛋白对McCoy细胞系的细胞毒性作用;液体培养基稀释法检测Vp1蛋白对大肠杆菌BL21和DH5α的抗菌作用.结果 衣壳蛋白Vp1对Q标准株E和D型的抑制率分别为78％和72％,对Ct临床野生株的抑制率为40％～70％,透射电镜结果显示Vp1蛋白能够抑制Ct的正常发育,使包涵体内出现异常增大的网状体.而该Vp1蛋白对非衣原体的其他生物体如大肠杆菌BL21、DH5α和真核细胞McCoy的生长没有影响.结论 噬菌体衣壳蛋白Vp1对Ct的生长具有明显的抑制作用,为临床上Ct感染的治疗提供了新的思路.     

生殖道支原体、沙眼衣原体、加德纳菌感染与男性不育的关系

男性不育不是一种独立的疾病,而是多种疾病或多种因素造成的结果。为了解生殖道感染与男性不育的关系,本文对本院就诊的435例男性不育症患者及368例体检者进行支原体(Mycoplasma,Uu)、沙眼衣原体(Cblamydia,Ct)及阴道加德纳菌(Gardnerella vaginalis,Gv)检测,比较两组对象阳性率的差异,现报道如下。     

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.     

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 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.     

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.