苯丙胺对大鼠学习能力和海马结构GFAP阳性细胞结构的影响

目的观察苯丙胺对大鼠学习能力、GFAP免疫阳性细胞结构和亚细胞结构的影响。方法苯丙胺腹腔注射SD大鼠,用水迷宫检测其空间辨别性学习能力,用免疫组织化学方法观察海马结构胶质纤维酸性蛋白(GFAP)免疫阳性细胞的形态变化;用电镜方法检测GFAP免疫阳性细胞超微结构变化。结果学习能力检测发现,在14d以前,苯丙胺组大鼠较生理盐水组的平均运行时间和平均潜伏期缩短(P0.05);在15～28d,苯丙胺组大鼠较生理盐水组正确率降低(P0.05);在29～42d,苯丙胺组大鼠较生理盐水组正确率降低、平均运行时间延长(P0.05)。GFAP阳性细胞形态学观察发现,正常对照组大鼠海马结构星形胶质细胞主要分布在海马以及齿状回的多形层和分子层;生理盐水组大鼠海马结构星形胶质细胞分布与游水组相似,但细胞突起变长及增粗,分支增多,且各亚区比正常对照组相应亚区的星形胶质细胞数量增多(P0.05);苯丙胺组大鼠海马星形胶质细胞分布与正常对照组相似,但细胞突起变长、增粗和分支增多更明显,各亚区的星形胶质细胞比正常对照组和生理盐水组相应亚区增多(P0.05);电镜观察发现苯丙胺组大鼠脑内星形胶质细胞增生肥大,亦可见退变星形胶质细胞。结论在本实验条件下,苯丙胺导致大鼠空间辨别性学习记忆能力下降,引起大鼠海马结构星形胶质细胞增生和损害。     

雷公藤内酯醇(T10)通过抑制脊髓内胶质细胞的激活改善大鼠CFA诱导的炎性痛的机制研究

目的:观察腹腔注射雷公藤内酯醇(Triptolide,T10)对完全弗氏佐剂(complete Freund’s adjuvant,CFA)诱导的大鼠慢性炎性痛行为的改善作用并探讨其机制。方法:SD大鼠随机分为四组,即Saline+Veh组、Saline+T10组、CFA+Veh组和CFA+T10组。后两组大鼠于右侧足底注射CFA制备大鼠慢性炎性痛模型,前两组则在相同部位注射生理盐水。第二组和第四组大鼠分别于造模前1 h给予T10腹腔注射,随后每12 h给予腹腔注射药物1次,持续用药7 d;第一组和第三组则以相同方式给予100 ml/kg的生理盐水作为对照。采用辐射热法和机械刺激法连续观察给药后大鼠的痛行为;应用免疫组织化学染色和Western Blot方法观察连续给予T10 3d和7 d后大鼠腰膨大平面脊髓背角内小胶质细胞和星形胶质细胞的活化程度;应用实时定量PCR(RT-PCR)方法观察连续给药7 d后大鼠腰5背根神经节内炎性因子,如白细胞介素-6(interleukin-6,IL-6)、白细胞介素-1β(interleukin-1 beta,IL-1β)和肿瘤坏死因子-α(tumour necrosis factor-α,TNF-α)的表达变化。结果:(1)与CFA+Veh组相比,CFA+T10组大鼠机械缩足阈值及辐射热痛潜伏期显著下降(P0.05),并持续到造模成功后7 d,表明腹腔注射T10可以明显改善CFA诱导的大鼠机械痛敏和辐射热痛敏。(2)免疫组织化学染色和Western Blot结果显示:造模后3 d和7 d后CFA+Veh组大鼠脊髓背角出现大量CFA诱导活化的小胶质细胞和星形胶质细胞;小胶质细胞标记物IBa-1和星形胶质细胞标记物GFAP的表达量分别在CFA造模后3 d和7 d显著增高(P0.05),而给予T10 3 d和7 d后炎性痛大鼠腰膨大平面IBa-1和GFAP的活化程度均显著减轻(P0.05)。(3)RT-PCR结果显示:与Saline+Veh组相比,CFA造模7 d后腰5背根神经节内炎性因子IL-1β、IL-6 mRNA和TNF-αmRNA的表达显著增高(P0.05),腹腔注射T10 7 d后上述炎性因子的表达则显著降低(P0.05)。结论:腹腔注射T10可以显著改善CFA诱导的大鼠慢性炎性痛,其机制可能与抑制脊髓背角小胶质细胞和星形胶质细胞的活化以及炎性因子IL-1β、IL-6和TNF-α的合成有关。     

雷公藤多甙对大鼠海马内注射β-淀粉样蛋白后星形胶质细胞增生的影响

目的：探讨大鼠海马内注射v淀粉样蛋白1-40(Aβ1-40)后海马星形胶质细胞表达GFAP的变化及雷公藤多甙(TWP)对其的影响。方法：在大鼠左侧海马定向注射Aβ1-40作为Aβ组，注射生理盐水作为对照组，雷公藤多甙腹腔注射治疗海马内注入了Aβ1-40的大鼠为TWP组。用免疫组织化学方法观察各组大鼠海马星形胶质细胞GFAP的表达。结果：Aβ组海马星形胶质细胞增生、肥大，GFAP阳性细胞数增多，细胞截面积明显增大。TWP组星形胶质细胞数较Aβ组减少，并且细胞截面积明显缩小。结论：雷公藤多甙能抑制Aβ诱导的海马星形胶质细胞的反应性增生。     

8-O-乙酰山栀子苷甲酸通过抑制脊髓背角JNK的磷酸化水平改善大鼠炎性痛的研究

目的:探究8-O-乙酰山栀子苷甲酸(8-O-acetyl-SM,8-Oa S)对于慢性炎性痛模型大鼠痛行为及脊髓背角星形胶质细胞内c-Jun氨基酸末端激酶(c-Jun N-terminal kinase,JNK)磷酸化水平的影响。方法:运用大鼠足底注射完全弗式佐剂(complete Freund’s adjuvant,CFA)的方法建立慢性炎性痛模型;通过腹膜腔注射8-Oa S进行干预;采用von Frey丝测定大鼠足底50%机械性缩足反射阈值;应用免疫荧光组织化学染色法观察大鼠腰膨大节段脊髓背角胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)及磷酸化的JNK(phosphorylatedc-Jun N-terminal kinase,p JNK)表达;应用Western Blot方法对大鼠脊髓背角内GFAP、JNK以及p-JNK的表达水平进行定量分析。结果:(1)行为学结果显示:与对照组相比,CFA模型大鼠机械性痛阈明显降低(P0.01),腹膜腔注射8-Oa S可有效提高CFA诱导的机械性痛阈值(P0.05);(2)免疫荧光染色结果显示:CFA模型脊髓背角内GFAP的表达量明显高于正常组,且p JNK基本表达于星形胶质细胞内;(3)Western Blot结果显示:与对照组相比,CFA造模后7 d脊髓背角内GFAP和p JNK表达明显上调,腹膜腔给予8-Oa S可以显著下调CFA诱导的脊髓背角内GFAP和p JNK的水平(P0.01)。结论:腹膜腔内给予8-Oa S可有效提高炎性痛大鼠的机械性痛阈,其机制是通过下调脊髓背角星形胶质细胞内JNK信号通路的磷酸化水平进而抑制星形胶质细胞的激活。     

脊髓星形胶质细胞参与福尔马林诱导的炎性痛

目的观察脊髓星形胶质细胞是否参与了福尔马林诱导炎性痛的过程。方法采用福尔马林制备炎性痛模型,生化分光光度法检测小鼠脊髓后角谷氨酸浓度,蛋白印迹法观察脊髓后角星形胶质细胞、谷氨酸转运体1、谷氨酸天冬氨酸转运体和谷氨酰胺合成酶蛋白表达水平。结果炎性痛模型组小鼠脊髓后角谷氨酸含量,星形胶质细胞数量,谷氨酸天冬氨酸转运体,谷氨酸转运体1和谷氨酰胺合成酶表达水平明显高于对照组。结论脊髓星形胶质细胞可能参与福尔马林诱导炎性痛的过程。     

海马内注射LPS后诱导大鼠皮质星形胶质细胞活化和PirB的表达

目的:探讨海马内注射LPS(lipopolysaccharide,LPS)后,大鼠皮质神经元和星形胶质细胞PirB(paired-immunoglobulin-like receptor B)的表达变化。方法:成年SD大鼠,分为对照组和实验组,用免疫组织化学法检测大鼠脑皮质PirB的表达及星形胶质细胞GFAP的表达变化,免疫荧光双重标记技术,观察星形胶质细胞与PirB阳性细胞的共存。结果:PBS注射的对照组动物脑皮质Ⅴ层内可见PirB阳性神经元样细胞,呈圆形、卵圆形和锥体形,免疫反应产物呈棕色、主要位于细胞膜上;海马内注射LPS 30 d后,PirB阳性神经元样细胞和PirB阳性神经胶质样细胞主要分布于皮质Ⅳ、Ⅴ层,免疫反应产物位于胞质和突起内;另外,可见大鼠梨状皮质、内嗅皮质内GFAP阳性星形胶质细胞明显被激活,表现为细胞胞体相对增大,突起变粗。PirB分别和MAP-2、GFAP、CD11b免疫荧光双重标记染色显示:PirB和MAP-2阳性神经元的胞体存在共定位;PirB与GFAP阳性染色存在部分共定位,但未见PirB与CD11b阳性染色双标细胞。结论:LPS能诱导大鼠皮质星形胶质细胞活化和PirB蛋白表达上调,而且部分活化的星形胶质细胞表达PirB,PirB可能参与脑内炎症突触可塑性改变和学习记忆功能缺失的免疫调节。     

雷公藤甲素对LPS诱导的神经炎症中星形胶质细胞和小胶质细胞激活的抑制作用

选用健康成年雄性SD大鼠,用不同剂量的雷公藤甲素[10、25、50μg/(kg·d)]预处理5d后,海马注射内毒素(LPS,4μg)24h。采用免疫组织化学和荧光组织化学方法观察海马内星形胶质细胞标记物胶质纤维酸性蛋白(GFAP)和小胶质细胞标记物蓖麻凝集素-1(RCA-1)表达的变化。结果显示:与生理盐水对照组比较,LPS注射引起注射部位GFAP表达增加,RCA-1阳性细胞增多、变大。而雷公藤甲素[50μg/(kg·d)]可明显下调LPS诱导的GFAP和RCA-1的表达,其抑制程度与药物剂量呈正相关。本研究结果提示,雷公藤甲素对海马内LPS诱导的星形胶质细胞和小胶质细胞的激活有明显的抑制作用。     

大鼠生后中枢神经系统S100B和胶质纤维酸性蛋白表达的变化

目的 探讨SD大鼠生后中枢神经系统发育过程中S100B和胶质纤维酸性蛋白(GFAP)的表达变化.方法 24只雄性SD大鼠分为生后7d、14d、21d和成年4组,用免疫组织化学方法对脑、脊髓切片进行S100B、GFAP抗体染色,观察不同时间点不同部位中两种阳性细胞平均数.结果 生后7d到成年,前额皮质、海马、纹状体、黑质和脊髓S100B阳性标记的密度和数量逐渐减少,生后2～3周时渐趋于稳定;脑内GFAP阳性星形胶质细胞(AST)随年龄增大逐渐增多,突起增粗增长,生后21d GFAP阳性细胞数量已接近成年;相反,脊髓GFAP阳性标记数则随年龄增长呈现由多到少的趋势;海马CAl区生后各年龄段GFAP和S100B免疫荧光双标显示,生后1周至成年S100B阳性细胞的数量明显减少,尤以分子层明显;随年龄增长,双标阳性细胞的比例逐渐增高,多集中分布于锥体细胞层和多形层.结论 大鼠中枢神经系统中S100B和GFAP两种星形胶质细胞蛋白存在不同的表达模式;同时S100B和GFAP蛋白的表达在发育过程中可能受不同机制的调节,并可能代表星形胶质细胞的不同亚型.     

鞘内注射甘珀酸降低CX43的表达对大鼠镜像痛的影响

目的:研究大鼠脊髓星形胶质细胞CX43的表达变化及对镜像痛的影响。方法:96只SD大鼠随机分为四组:Sham组,SNI组,SNI+NS组,SNI+CBX组,每组24只。各组分别于术前l d、术后3、7、10、l4和21 d观察大鼠疼痛行为学变化,其中Sham组,SNI组于术后14 d采用免疫组化法分析缝隙连接蛋白43(Cx43)、胶质原纤维酸性蛋白(GFAP)的表达情况;SNI+CBX组于术后第5~9 d鞘内注射甘珀酸10μl(20μg),SNI+NS组注射等量生理盐水,并于术后10、14和21 d三个时间点采用Western Blot的方法检测脊髓内CX43蛋白表达情况,采用免疫组化法分析GFAP的表达变化情况。结果:SNI组与Sham组相比,行为学上大鼠双侧机械缩足阈值降低,脊髓星形胶质细胞CX43及GFAP表达增加(P0.05);SNI+CBX组与SNI+NS组相比,Western Blot显示CX43表达降低,免疫组化显示脊髓背角双侧GFAP表达降低(P0.05),术后14 d,21 d大鼠双侧机械缩足阈值升高。结论:鞘内注射甘珀酸降低CX43的表达可抑制星形胶质细胞的活化,并缓解大鼠双侧的机械痛。     

卡马西平对大鼠海马星形胶质细胞形态和GFAP表达的影响

目的观察治疗剂量卡马西平对SD大鼠海马星形胶质细胞形态和胶质纤维酸性蛋白(GFAP)表达的影响。方法治疗剂量卡马西平每24 h单次灌胃给药后,于不同时间点经心脏灌注,取脑;GFAP免疫组化染色测量海马GFAP阳性细胞数量及形态。结果1周组海马CA1区星形胶质细胞的形态及GFAP表达无变化,1个月组星形胶质细胞数量及GFAP表达量无明显增加,3个月组GFAP阳性细胞数显著增加,达对照组的1.74倍(P<0.05);并伴细胞体积增大,侧支增多。结论治疗剂量的卡马西平对星形胶质细胞形态和GFAP表达的影响呈时间依赖性。     

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.