小鼠出生后肾脏发育过程中的细胞增殖与凋亡

目的：观察小鼠出生后肾脏发育过程中增殖细胞核抗原(PCNA)的表达及细胞凋亡的特征,探讨出生后小鼠肾脏发育过程中细胞增殖与凋亡的规律及其关系。方法：应用免疫组织化学技术和原位末端标记法(TUNEL法)分别检测小鼠出生后1～70d肾脏中PCNA阳性的细胞和凋亡细胞。结果：小鼠出生后1～70d,皮质中的肾小体、肾小管、髓放线以及髓质中的肾小管和集合管的细胞,早期增殖活跃,随着肾脏发育成熟而表达逐渐减弱。同时,也存在着细胞凋亡现象,且凋亡高峰一般出现在增殖高峰之后。结论：细胞增殖与凋亡在小鼠生后肾脏发育的整个过程中普遍存在,生后1～7d细胞增殖旺盛,增殖高峰之后出现凋亡高峰,生后28～70d两者活动均减弱。     

发育期小鼠肾小体细胞凋亡的研究

目的 研究小鼠肾小体发育过程中的细胞凋亡规律及形态学特点。方法 应用光镜、电镜技术和脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记法(TUNEL法)分别对不同胚龄和生后日龄小鼠肾小体内细胞凋亡进行观察。结果 胚龄14d肾小体发生时就出现细胞凋亡，胚龄18d左右达到高峰，l随后缓慢下降。肾小体内凋亡细胞以内皮细胞和足细胞多见。凋亡细胞的两种结局：1．被邻近的细胞所吞噬；2．落人肾小体的毛细血管腔或肾小囊中。结论 小鼠肾小体的整个发育过程均存在细胞凋亡现象，肾小体内细胞凋亡与肾小体发育的完善有关。     

增殖细胞核抗原在小鼠生后肾脏发育中的表达

目的：观察小鼠出生后肾脏发育过程中增殖细胞核抗原（PCNA）的表达特征,探讨出生后小鼠肾脏发育过程中细胞增殖的规律。方法：应用免疫组织化学技术检测小鼠出生后1、3、7、14、21、28和70d肾脏PCNA的表达。结果：小鼠出生后1～70d,皮质中的生肾区、肾小体、肾小管、髓放线以及髓质中的肾小管和集合管PCNA阳性细胞的表达具有一定的规律,早期PCNA阳性表达丰富,随着肾脏发育逐渐成熟而表达减弱直至消失。在70d成年小鼠肾脏中,没有检测到PCNA阳性细胞。结论：出生后小鼠肾脏皮质中的生肾区、肾小体、肾小管、髓放线以及髓质中小管的细胞增殖规律是由高逐渐降低的,直至成年完全停止。     

db/db自发性糖尿病小鼠睾丸生殖细胞增殖及凋亡研究

目的　研究糖尿病对生殖细胞增殖和凋亡的影响。　方法　以免疫组织化学ABC法检测增殖细胞核抗原 (proliferationcellnucleusantigen ,PCNA)检测生殖细胞的增殖情况 ,电镜和脱氧核糖核酸末端转移酶介导的dUTP缺口末端标记法 (TUNEL)检测凋亡的生殖细胞。　结果　PCNA阳性率随糖尿病病程进展 (糖尿病组 )和年龄增加 (对照组 )均呈明显下降趋势 ,但每年龄段糖尿病组PCNA阳性率均明显低于相应正常对照组。凋亡管横断面数及凋亡阳性率糖尿病组均较正常高 ,而且随糖尿病病程进展呈明显增加趋势 ,对照组中随加龄二者有增加。电镜下 ,糖尿病组小鼠睾丸精原细胞、精母细胞和精子细胞均可见到核染色质凝集、边集 ,凋亡小体形成和降解等凋亡过程。　结论　db/db自发性糖尿病小鼠睾丸生殖细胞PCNA表达减弱 ,TUNEL标记凋亡细胞增加 ,说明糖尿病使生殖细胞增殖与凋亡的平衡破坏 ,这可能是糖尿病生殖功能障碍的原因     

血管紧张素Ⅱ对发育期小鼠肾小体细胞增殖和凋亡的影响

目的 观察血管紧张素Ⅱ(AngⅡ)对体外培养E12d小鼠肾组织各期肾小体细胞增殖和凋亡的影响.方法 孕12d的母鼠40只随机分为6组,体外培养对照组(0mol/L AngⅡ)和施加不同浓度AngⅡ(10-9～10-5mol/L)组.应用免疫组织化学、原位缺口末端标记(TUNEL)法、免疫荧光双标技术及激光扫描共焦显微镜,观察体外培养2、4、6d后小鼠肾组织中各期肾小体增殖细胞核抗原(PCNA)和凋亡阳性细胞的表达. 结果 随培养浓度增高和培养时间延长,各期肾小体细胞增殖和凋亡指数均有增加,且增殖指数在AngⅡ低浓度(10-9、10-8、10-7mol/L)、短时间(培养2、4d)时变化明显;凋亡指数在高浓度(10-6、10-5mol/L)、长时间(培养6d)时变化明显. 结论 AngⅡ对发育期小鼠肾小体细胞有促增殖和凋亡作用,且此作用有浓度、时间依赖性.     

大鼠提睾肌缺血再灌注损伤时血管平滑肌细胞和内皮细胞的增殖与凋亡

目的研究大鼠提睾肌缺血再灌注后血管平滑肌细胞和内皮细胞的增殖和凋亡。方法以免疫组织化学SP法和末端脱氧核苷酸转移酶介导生物素标记法 (TUNEL)对大鼠提睾肌缺血再灌注后的血管平滑肌细胞和内皮细胞的Bcl-2、Bax、P53和增殖细胞核抗原 (PCNA)的表达进行观察研究。结果缺血再灌注后 ,血管平滑肌细胞和内皮细胞均有Bcl-2、Bax、P53和PCNA的表达 ,在平滑肌细胞 ,Bcl-2阳性细胞数量明显高于Bax和P53。PCNA阳性细胞明显增多。在内皮细胞 ,Bax和P53的表达最强 ,TUNEL阳性细胞率最高。结论大鼠提睾肌缺血再灌注可造成平滑细胞的增殖和内皮细胞的凋亡 ,其结果可能与微循环障碍有关。     

小鼠肾脏发育中的细胞凋亡

目的：研究小鼠肾脏发育过程中的细胞凋亡规律及形态学特点,方法：应用光镜、电镜技术和TUNEL法分别对不同胚龄、生后日龄小鼠肾脏细胞凋亡进行了观察。结果：皮质凋亡细胞多出现在生肾区S小体之间和是肾小体内,凋亡高峰期在胚龄14－18d之间,髓质凋亡细胞出现在肾小鼠管上皮内,凋亡高峰期在生后7d 左右。超微结构观察可见皮质和髓质凋亡细胞主要表现为核固缩,染色质凝集,细胞皱缩。皮质和髓质凋亡细胞结局为,被邻近细胞吞噬,或脱落到肾小管腔内,结论：小鼠肾脏发育过程中确有细胞凋亡;皮质中细胞凋亡与生肾区的出现和肾小体发育完善有关,髓质中细胞凋亡与髓质中肾小管和集合小0管的有发育完善有关。     

饮酒对小鼠睾丸生精小管一氧化氮合酶、增殖细胞核抗原表达及细胞凋亡影响的研究

目的　探讨酒精对小鼠睾丸的组织结构、内皮型一氧化氮合酶 (eNOS)、增殖细胞核抗原 (PCNA)及细胞凋亡的影响。　方法　用 5 %、10 %及 15 % 3种不同浓度的酒精作用于 2 2d龄小鼠 ,取睾丸做石蜡切片、HE染色 ;用免疫组织化学方法检测睾丸eNOS、细胞增殖的变化 ;TUNEL法检测细胞凋亡的变化 ,并进行统计学分析。　结果　随着酒精浓度的增大 ,睾丸组织结构发生明显改变 ,生精小管的直径逐渐减小 ,eNOS阳性细胞面积密度逐渐增大 ,单位面积内PCNA阳性细胞和凋亡细胞数目增加 ,高浓度酒精组与其他组差异极显著 (P <0 0 1)。　结论　酒精可使生精小管的直径变小 ,eNOS及PCNA表达增强 ,凋亡细胞增加并随酒精浓度的增大而变化加重。这可能是过量饮酒导致生精细胞减少 ,生殖能力降低的重要因素。     

遗传性视网膜变性rd小鼠及其感光细胞凋亡研究

目的 研究遗传性视网膜变性rd小鼠感光细胞层的发育变化及细胞凋亡。方法 对出生后5d到40d的rd小鼠及对照小鼠视网膜感光细胞层进行光镜及超微结构观察、TUNEL法检测及形态计量学分析。结果 与同龄对照鼠相比,rd小鼠出生后第10d视网膜开始变性,尔后1周内感光细胞迅速减少,第18d时只残留一层视椎细胞。rd小鼠出生后第10d感光细胞层开始出现TUNEL染色阳性细胞,第14d及16d达到高峰。电镜下变性高峰期rd小鼠视网膜感光细胞层可见大量浓缩核、染色质边聚及凋亡小体。结论 rd小鼠视网膜感光细胞在发育过程中变性,并通过凋亡的方式死亡。     

大鼠视网膜发育过程中细胞增殖与凋亡的形态学

目的:观察视网膜发育过程中增殖细胞核抗原(PCNA)的表达变化及细胞凋亡状态。方法:14~20 d胎鼠(E14-20 d)、生后0~15 d幼鼠(P0-15)及成年鼠(P36),取出眼球组织,免疫组织化学显色及原位末端标记(TUNEL)观察PCNA的表达和细胞凋亡变化。结果:发育早期E14~P7,可观察到有较强PCNA阳性表达,P9时PCNA阳性细胞数目明显减少,P15开始未见明显PCNA阳性表达。E18可观察到凋亡细胞,随着发育的进行,凋亡细胞逐渐增多,P7时凋亡细胞数目最多,之后又逐渐减少,成熟的视网膜组织内未见明显凋亡细胞。结论:在视网膜神经细胞发育过程中,细胞的增殖和凋亡呈现了有序的动态变化,正是这种有序的增殖和凋亡的平衡才使视网膜最终发育成了正常的组织结构及功能。     

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 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.