Fmr1基因在大鼠快速眼动睡眠剥夺后脑皮质、海马和丘脑区的表达

目的探讨快速眼动(REM)睡眠剥夺过程中Fmr1基因在大鼠皮质、海马和丘脑区的表达及变化。方法采用改良多平台水环境法(MMPM)制作大鼠睡眠剥夺模型,采用免疫组织化学法及RT-PCR方法检测Fmr1基因的表达变化。结果在皮质和丘脑中,与CC组和TC组相比,SD1d和SD2d组的Fmr1基因表达无明显变化,SD3d组开始增高(P0.05),SD5d、SD7d组和SD9d组显著增高(P0.01);在海马中,与CC组和TC组相比,SD1d和SD2d组的Fmr1基因表达无明显变化,SD3d组开始降低(P0.05),SD5d、SD7d组和SD9d组显著降低(P0.01)。结论 Fmr1基因在大鼠睡眠剥夺第3天开始表达发生变化,在皮质和丘脑中表达增高,在海马中表达降低。     

慢性吗啡处理及戒断后大鼠前额皮质、海马和杏仁核中Parvalbumin的表达变化

为了观察慢性吗啡处理及戒断后大鼠前额皮质、海马和杏仁核中parvalbumin(PV)的表达变化,为其功能的研究提供形态学依据,本实验将30只雄性SD大鼠随机分为吗啡依赖组和生理盐水对照组。吗啡依赖组大鼠腹膜腔注射吗啡,2次/d,起始剂量为5mg/kg,逐日递增5mg,至第10d为50mg/kg;对照组注射同体积的生理盐水。于末次注射后动物分别存活3h、3d和14d。用免疫组化方法和相对平均灰度值检测前额皮质、海马和杏仁核内PV的表达。结果显示:在生理盐水处理组各存活时间点,前额皮质、海马和杏仁核内PV的表达相同。和生理盐水对照组相比,3h时海马和杏仁核内PV的表达明显增加(P<0.05),但前额皮质内PV的表达减少。第3d时,海马CA1、CA3、CA4、齿状回和杏仁核内PV的表达减少,但CA2区PV表达继续增加,而前额皮质的表达开始恢复。至第14d时,CA2区PV的表达开始恢复,但CA1、CA4和杏仁核内PV的表达又开始增加,明显高于第3d组(P<0.05)。以上结果提示慢性吗啡处理及戒断后PV的表达具有区域特异性和时相特异性;这种变化在戒断早期可能主要与躯体依赖相关,而戒断晚期主要与精神依赖相关。     

脂肪来源干细胞移植促进大鼠脑梗死周边区皮质结直肠癌缺失蛋白(DCC)和神经丝蛋白200(NF-200)的表达

目的研究脂肪来源干细胞(ADSC)移植对大鼠脑梗死周边区皮质中结直肠癌缺失蛋白(DCC)和神经丝蛋白200(NF-200)表达的影响。方法 SD大鼠随机分为假手术组、模型组和ADSC组。采用改良Longa线栓法制备大鼠大脑中动脉闭塞脑梗死模型。ADSC组大鼠于造模后24 h经颈动脉注射入50μL用PKH-26标记的ADSC悬液(含约2×10~6个细胞),模型组大鼠注射等剂量的生理盐水。于造模后第7、 14天,在荧光显微镜下观察脑组织中ADSC的迁移情况,采用Western blot法检测脑梗死周边区皮质DCC、 NF-200的蛋白水平,免疫荧光法检测脑梗死周边区皮质中DCC、 NF-200的表达和分布。结果 ADSC组大鼠脑组织中可检测到PKH-26标记的ADSC,且大部分位于脑梗死周边区皮质中。造模后第7、 14天,模型组DCC的表达均高于假手术组,而NF-200的表达均低于假手术组; ADSC组DCC和NF-200的表达在各个时间点均高于模型组。在梗死周边区皮质, DCC主要表达于NF-200阳性的神经元轴突中。结论经颈动脉移植的ADSC可以迁移至脑梗死周边区皮质,并可能通过促进该区域DCC、 NF-200的表达,从而强化神经轴突的再生修复。     

睡眠剥夺对HSP70表达及脑超微结构的影响

目的：观察睡眠剥夺（SD）后大鼠脑组织HSP70表达的变化及对超微结构的影响。方法：44只雄性SD大鼠随机分为11组，每组4只，免疫组织化学方法检测HSP70的表达，电镜观察海马超微结构的变化。结果：睡眠剥夺后12小时即可在大脑皮质及海马观察到HSP70阳性细胞，2—3天数量达到高峰，7天时明显下降。白天睡眠剥夺12小时（SDd12h）组HSP70阳性细胞数较夜晚睡眠剥夺12小时（SDn12h）组多（P〈0．05）。RS组大脑皮质HSP70阳性细胞数较白天睡眠剥夺1天（SDd1d）组减少（P〈0．05）。白天睡眠剥夺3天（SDd3d）海马出现超微结构改变，白天睡眠剥夺7天（SDd7d）后改变更加明显。结论：睡眠剥夺可影响大鼠脑组织HSP70表达及超微结构。     

不同时长REM期睡眠剥夺及莫达非尼干预后大鼠下丘脑Orexin A神经元的表达

目的:探讨不同程度快动眼睡眠(REM)期睡眠剥夺(sleep deprivation,SD)及莫达非尼干预后大鼠下丘脑Orexin A神经元的表达。方法:将成年雄性Sprague-Dawley大鼠随机分为和SD组和对照组,SD组又分为用药组(drug group,DG)和非用药组(non-drug group,NDG),每组分SD12,24,48,72,96h共5个小组;对照组(cage control,CC)1个小组,正常饲养于笼中。每小组3只大鼠。采用小平台水环境法建立大鼠REM期SD模型。免疫组化方法观察大鼠下丘脑Orexin A阳性神经元的数量。结果:Orexin A阳性表达在SD12,24h时长的DG组与NDG组表达较CC组均有增加(P0.05)而二者之间差别不明显(P0.05);在SD48,72,96h时长的NDG组的表达较CC组下降(P0.05),而DG组和CC组间无显著差异(P0.05)且明显高于NDG组表达(P0.05)。结论:推测莫达非尼可能是通过活化下丘脑促觉醒肽Orexin A的分泌和表达实现促觉醒作用。     

慢性复合应激对大鼠海马生长休止蛋白7表达的影响

目的 探讨慢性复合应激对大鼠海马神经元中生长休止蛋白7(Gas7)的表达变化及意义.方法 36只大鼠随机分为慢性复合应激组和正常对照组.复合应激组动物进行6周的垂直旋转、睡眠剥夺、捆绑(6h/d)和夜间光照等慢性复合性应激试验;实验结束后,所有动物采用免疫组织化学和免疫印迹等方法 检测大鼠海马Gas7蛋白表达的变化和神经元凋亡情况.结果 Gas7在对照组和实验组大鼠海马各区均有表达,主要表达在海马神经元的胞质和突起内.其中在CA1区和齿状回呈较强阳性表达,CA3区表达则较弱;慢性复合应激组CA1区和齿状回阳性染色加深,平均吸光度明显上升(P<0.05),而CA3区则不明显(P>0.05).慢性复合应激组中部分切片在下托可见少量Caspase-3免疫反应阳性细胞.免疫印迹检测表明,慢性复合应激组大鼠海马Gas7表达明显增强,与正常对照组相比,差异具有显著性(P<0.05).结论 Gas7可能参与了在应激情况对神经元的保护作用,以及促进神经元的发生和发育.     

缺氧缺血性脑损伤新生大鼠海马区脑红蛋白、凋亡相关蛋白Bcl-2、Bax的表达及其相关性

目的:探讨新生大鼠缺氧缺血性脑损伤(HIBD)后海马CA1区脑红蛋白(Ngb)及凋亡相关蛋白Bcl-2、Bax的变化规律及其相关性。方法:选用60只新生7日龄Wistar大鼠,随机分为三组:sham组(n=6),正常对照组(n=6)和HIBD组(n=48)。HIBD组新生大鼠建立HIBD模型,分别于HIBD后3h、6h、12h、24h、48h、72h、7d、14d取材,应用免疫组织化学染色方法检测各组海马CA1区Ngb及Bcl-2、Bax的表达变化情况。结果:(1)与sham组相比,HIBD组Ngb阳性细胞数于缺氧缺血后3h开始下降,但差异无统计学意义;6h、12h、24h呈逐渐下降趋势,阳性细胞数量已明显低于sham组;到48h时达最低水平;14d时Ngb阳性细胞数量基本恢复至sham组水平。(2)与sham组相比,HIBD组于缺氧缺血3hBcl-2阳性表达细胞开始显著增多,于12h达到高峰,24h时开始逐渐下降,14d时略高于sham组表达,但差异无统计学意义。(3)与sham组相比,HIBD组Bax蛋白免疫反应阳性细胞数目于缺氧缺血后3h时开始增加,12h时明显增多,于48h达高峰,此后逐渐下降,7d时阳性细胞数基本恢复至sham组水平,14d时未见Bax蛋白的阳性表达。(4)新生大鼠海马CA1区内,Ngb阳性细胞数与Bcl-2阳性细胞数无相关;与凋亡相关蛋白Bax阳性细胞数呈负相关(r=-0.697,P0.01)。结论:Bcl-2与Bax基因参与了HIBD后神经细胞凋亡的发生;Ngb可能通过抑制促凋亡蛋白Bax的表达,从而减少神经细胞的凋亡,发挥其神经保护作用。     

丹参酮ⅡA对脑缺血再灌注大鼠顶叶皮质TNF-α和NF-κB表达的影响

目的探讨丹参酮ⅡA对局灶性脑缺血再灌注大鼠顶叶皮质肿瘤坏死因子-α(TNF-α)和核转录因子-κB(NF-κB)表达的影响。方法选取36只健康雄性大鼠,随机分为假手术组(Sham,12只),缺血再灌注组(I/R,12只)和丹参酮ⅡA治疗组(TanⅡA,12只),用线栓法阻塞大鼠右大脑中动脉制作局灶性脑缺血再灌注模型,脑缺血2h再灌注24h后,应用免疫组织化学方法与Western blot方法检测大鼠手术侧顶叶皮质TNF-α和NF-κB的表达。结果与Sham组相比,I/R组大鼠顶叶皮质TNF-α与NF-κB蛋白的阳性表达的平均光密度值显著升高(0.05);而与I/R组相比,TanⅡA组大鼠顶叶皮质TNF-α与NF-κB蛋白的阳性表达的平均光密度值显著降低(0.05)。结论丹参酮ⅡA能够降低脑缺血再灌注大鼠顶叶皮质TNF-α与NF-κB蛋白的表达水平。     

脑震荡鼠大脑多巴胺转运体变化研究

为了进一步探讨脑震荡(cerebral concussion,CC)大鼠认知行为障碍的神经生物学基础,本实验观察了CC大鼠多巴胺转运体(dopamine transporters,DAT)的变化规律。用自制金属单摆式闭合性脑损伤打击装置复制脑震荡大鼠模型48只,随机分为伤后1、2、4、8、16、24d六个实验组,另设一正常对照组(n=8)。运用抗DAT抗体SP法行免疫组化染色,并测定大脑前额叶皮质(PFC)、尾壳核(CPu)、伏隔核(Acbc)、内侧隔核(MS)、斜角带垂直臂核(VDB)、斜角带水平臂核(HDB)、梨形皮质(Pir)、顶叶皮质(CPL)、海马CA1-3、丘脑中央背内侧核(MD)、丘脑背外侧核(LD)、下丘脑腹内侧核(VMH)和杏仁核等16个脑区DAT阳性表达物的光密度值。结果表明,与正常对照组相比脑震荡后大鼠多个脑区DAT阳性表达物呈下调变化。PFC、CPu-2、MS、LD、MD的DAT阳性表达物在伤后第2d表达最低;CPu-1、VDB、HDB、CPL、CA1、CA2、CA3、VMH和杏仁核的DAT阳性表达物在伤后第4d表达最低;Pir的DAT阳性表达物在伤后第8d表达最低;Acbc的DAT阳性表达物在伤后第16d表达最低。各脑区恢复时间各不相同,至24d基本恢复正常。以上结果提示DAT可在大鼠不同脑区广泛表达;脑震荡后其在多个脑区呈下调性改变,提示多巴胺转运体的伤后变化可能参与了脑震荡后的认知障碍。     

脂肪来源的干细胞移植对脑缺血大鼠神经轴突再生的影响

目的:探讨脂肪来源的干细胞(ADSC)移植对脑缺血大鼠神经轴突生长以及神经胶质酸性蛋白(GFAP)、神经突蛋白(Neuritin)、神经微丝蛋白200(NF-200)表达的影响。方法:54只清洁级成年雄性SD大鼠,随机分为3组:假手术组(Sham组)、模型组(MCAO组)及MCAO+ADSC治疗组(ADSC组),每组18只。采用改良Zea-Longa线栓制法大脑中动脉栓塞(MCAO)模型,ADSC移植前用DAPI标记,ADSC组于造模成功1 d后经侧脑室注射入ADSC(1×106),分别于术后7 d、14 d、28 d观察其恢复情况,并断头取脑,通过免疫荧光、Western blot法检测脑缺血组织中GFAP、Neuritin、NF-200表达情况。结果:ADSC组缺血周边区脑组织中能观察到DAPI染色的阳性细胞;ADSC组与MCAO组相比在各个时间点脑组织中GFAP阳性细胞表达明显降低(P<0.05),神经突蛋白和神经微丝蛋白200表达明显增高(P<0.05)。结论:ADSC移植后可引起脑缺血后期组织中Neuritin、NF-200有效表达,并抑制GFAP阳性细胞增生,促进了神经轴突再生和修复。     

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.     

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.