GDNF在坐骨神经损伤后大鼠L4—6背根节神经元的表达及变化

目的 观察坐骨神经夹伤后不同时间点,胶质细胞源性神经营养因子（ＧＤＮＦ）在成年大鼠Ｌ４～６背根节（ＤＲＧ）中的分布,探讨ＧＤＮＦ对感觉神经元损伤的反应。方法 成年大鼠右侧坐骨神经夹伤后,分别取伤后１、５、７、１０、１４、２８和５６ｄＬ４～６背根节,行抗ＧＤＮＦ多克隆抗体免疫组织化学ＡＢＣ法染色,之后对染色结果进行图像分析。结果 ＧＤＮＦ免疫反应存在于坐骨神经夹伤后１、５、７、１０、１４、２８和５６ｄ成年大鼠Ｌ４～６背根节神经元中,图像分析结果表明,伤后１、３、５和７ｄ的损伤侧背根节神经元的平均光密度大于对照侧背根节神经元。结论 大鼠坐骨神经夹伤后,ＧＤＮＦ在背根节Ｌ４～６感觉神经元中有一定量的变化,且伤后１周内ＧＤＮＦ在伤侧背根节神经元中的表达增强。     

大鼠坐骨神经损伤后Src抑制的蛋白激酶C的底物在背根神经节中的表达变化

目的:探讨大鼠坐骨神经损伤后,Src抑制的蛋白激酶C的底物(SSeCKS)在背根神经节(DRG)中的表达变化及其意义。方法:制备成年SD(Sprague-Dawley)大鼠坐骨神经夹伤及切断模型。通过Western印迹法、Real-time PCR及免疫组织化学方法检测坐骨神经损伤后SSeCKS在DRG中表达的时空变化。结果:大鼠坐骨神经夹伤后6h,DRG中可检测到SSeCKS的表达并逐步升高,伤后12h达到高峰,2d后逐渐下降;而大鼠坐骨神经切断后DRG中SSeCKS的表达高峰发生在伤后2周,1d时最低;SSeCKS主要分布于DRG大、中、小神经元胞质;SSeCKS与NeuN、NF200以及GAP43存在共定位。结论:大鼠坐骨神经损伤后,引起DRG中SSeCKS的表达变化,其可能参与疼痛信号转导通路并与周围神经损伤后的再生有关。     

移植坐骨神经大鼠相应背根神经节中生长相关蛋白43的表达

背景:周围神经移植后,相应的背根神经节感觉神经元胞体内生长相关蛋白43(GAP-43)表达的时程与规律尚未明确。目的:观察大鼠坐骨神经移植术后GAP-43在相应背根神经节中的表达变化。方法:取健康成年雄性Wistar大鼠,随机分为2组,对照组行假手术组;实验组行坐骨神经移植,分别于术后3d,1,2,4,6,8周6个时间点处死后取其手术侧L4~L5背根神经节,应用RT-PCR和Western Blot技术检测GAP-43mRNA及其蛋白表达。结果与结论:对照组大鼠背根神经节中GAP-43mRNA表达量较低,且随时间无明显改变;实验组术后1周时GAP-43mRNA在相应背根神经节中即有表达增强,2周时达到高峰,6~8周表达逐渐减弱。两组大鼠背根神经节中GAP-43蛋白与GAP-43mRNA表达的变化规律相同。结果表明神经损伤后相应神经元的再生能力存在损伤反应性变化。     

5-HT_(1A)受体亚型mRNA在坐骨神经分支选择性损伤模型大鼠脊髓和背根节中的表达变化(英文)

坐骨神经分支选择性损伤 (SNI)模型是一种新型神经病理痛模型。本实验用 SD雄性大鼠 ,分支结扎并切断左侧坐骨神经干的胫神经和腓总神经 ,保留腓肠神经分支 ,右侧仅暴露坐骨神经。术后 1、2、3、4、7、14、2 1和 2 8d,用 RT-PCR的方法对 5 -HT1 A受体 m RNA在腰髓的背角和背根神经节 (DRG)的表达水平进行检测。结果显示 ,5 -HT1 A受体 m RNA在损伤侧腰髓背角内的表达水平于 1d后开始升高 ,7d时达高峰 ,随后逐渐下降 ,但仍高于正常水平。其表达水平在对侧脊髓背角内没有明显变化。在损伤侧 DRG内 ,5 -HT1 A受体 m RNA的表达水平于 1d后开始增高 ,4d时达高峰 ,随后开始下降 ,但仍维持较高的表达水平 ;而损伤对侧 DRG内的 5 -HT1 A受体 m RNA的表达没有变化。上述结果提示 5 -HT1 A受体亚型可能在脊髓及外周伤害性信息的传递和调节中发挥着重要作用 ,本研究的结果为进一步了解 5 -HT1 A受体在神经病理性痛中的作用机制提供了依据。     

坐骨神经慢性卡压损伤模型大鼠背根神经节纤维化改变

文题释义：组织纤维化：是结缔组织过度增生和细胞外基质沉积的结果,纤维化过程是一个异常的、不受控制的组织修复过程,其中组织损伤和自身免疫疾病是导致组织纤维化的主要因素,过度的组织纤维化最终导致组织结构和功能改变。 慢性神经卡压损伤：指周围神经在特定部位受到慢性卡压引起的相应神经功能障碍。 背景：既往的研究主要集中于周围神经慢性卡压损伤所导致的靶器官——骨骼肌萎缩及其纤维化发生的机制研究,关于周围神经慢性卡压损伤信号向上引起背根神经节功能改变的研究较少。 目的：观察大鼠坐骨神经慢性卡压损伤对背根神经节纤维化的影响。方法：按照Mackinnon设计的方法制备大鼠坐骨神经慢性卡压模型,术后3周,分别取大鼠坐骨神经卡压侧和对侧L_4-6背根神经节,采用RT-PCR、Western blot及免疫荧光检测大鼠卡压侧和对照侧背根神经节内转化生长因子β1、结缔组织生长因子及胶原蛋白Ⅰ的表达变化。结果与结论：①损伤3周后,相比于对照侧,背根神经节内转化生长因子β1、结缔组织生长因子及胶原蛋白Ⅰ的mRNA及蛋白表达均具有相同的升高趋势(P < 0.05);②损伤3周后,大鼠卡压侧和对照侧背根神经节内转化生长因子β1和结缔组织生长因子主要表达在背根神经元内和轴突内,而胶原蛋白Ⅰ主要表达在背根神经元和轴突的周围,形成包绕背根神经元和轴突的网状结构;③上述数据证实,大鼠坐骨神经慢性卡压损伤可以导致背根神经节纤维化的改变,并且背根神经节纤维化的改变可能与背根神经元内转化生长因子β1及结缔组织生长因子表达升高有关。ORCID: 0000-0003-1632-0837(黎琴文) 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

p27kip1和Skp2在大鼠坐骨神经夹伤后脊髓中的表达变化

目的:研究p27~(kip1)和Skp2在正常和坐骨神经夹伤后大鼠脊髓中的表达变化。方法:Westem印迹法,免疫组织化学,免疫荧光双标。结果:Western印迹结果显示坐骨神经夹伤后,脊髓中p27~(kip1)表达下降后又有所恢复,并且p27~(kip1)和Skp2在脊髓中的表达变化呈负相关。免疫组织化学及免疫荧光双标显示,p27~(kip1)和Skp2在正常和损伤后的脊髓神经元和胶质细胞中都有表达。坐骨神经夹伤后p27~(kip1)和Skp2在脊髓腹角阳性细胞数目变化呈负相关。结论:坐骨神经损伤后p27~(kip1)。和Skp2在脊髓腹角神经元及其周围的胶质细胞表达变化相关,对研究脊髓损伤和修复的分子机制有重要意义。     

5-HT受体亚型mRNAs在大鼠背根神经节和三叉神经节的表达—原位杂交组织化学法研究

应用原位杂交组织化学技术观察了大鼠背根节和三叉神经节内 5 -HT2 、5 -HT3 、5 -HT4、5 -HT5 、5 -HT6 和 5 -HT7受体亚型 m RNAs的表达。结果显示 :在背根节和三叉神经节中均观察到了 5 -HT2 A、5 -HT3 、5 -HT4和 5 -HT7受体亚型 m RNAs阳性细胞 ;两种神经节内上述受体亚型 m RNAs阳性细胞的百分比不同。 5 -HT3 受体亚型 m RNA阳性细胞的百分比最高 ,分别为 3 0 .4% (背根节 )和 3 2 .8% (三叉神经节 )。约 17.2 %和 14 .6 %的背根节和三叉神经节细胞分别呈 5 -HT2 A受体 m RNA阳性 ;而 5 -HT4和 5 -HT7受体亚型 m RNAs阳性细胞在此二节中的百分比分别为 14 .9%、2 4.7%和 11.2 %、9.8%。各受体亚型 m RNA阳性细胞均以中、小型节细胞为主。在各神经节内均未观察到 5 -HT2 B、5 -HT2 C、5 -HT5 和 5 -HT6 受体 m RNA的表达。上述结果提示 5 -HT2 A、5 -HT3 、5 -HT4和 5 -HT7受体亚型在外周伤害性感受中可能发挥着重要的作用     

大鼠坐骨神经夹伤后SSeCKS的表达变化

为了探讨大鼠坐骨神经夹伤后SSeCKS(Src抑制的蛋白激酶C底物)在神经中的表达变化及其意义,本研究制备了成年SD大鼠坐骨神经夹伤模型,通过Western blotting和免疫组织化学方法检测坐骨神经夹伤后SSeCKS表达的时空变化。Western blotting结果显示:大鼠坐骨神经夹伤后,SSeCKS的表达迅速升高,伤后6h即达到高峰,之后逐渐下降。免疫组织化学染色结果表明SSeCKS在神经夹伤两端高表达,以近端明显,呈簇状聚集。夹伤远端表达较近端稍低。远离夹伤处及相应对侧,SSeCKS的表达水平有所下降且分布较为均一。免疫荧光组织化学双标记结果显示:夹伤两端SSeCKS与S100和NF200的共存不明显,但可见部分SSeCKS与GAP43双标纤维。本研究提示大鼠坐骨神经夹伤可引起SSeCKS的表达变化,该变化可能参与周围神经损伤后某些伤害性刺激信号分子的转导并与损伤后神经的再生及功能修复有关。     

受体酪氨酸激酶Eph基因与肿瘤微环境及相关治疗的研究进展

Eph受体是目前发现人类最大的受体酪氨酸激酶家族,包括A类受体(Eph A)和B类受体(Eph B),其配体也包括两类(Ephrin A和Ephrin B)。与其他受体酪氨酸激酶基因一样,Eph基因在胚胎生长发育中发挥重要作用。近年大量研究发现其在肿瘤发生、进展和转移中也具有重要功能,但具体分子机制尚不明确。长期以来,人们研究肿瘤侵犯和转移的重点均集中在肿瘤细胞上,肿瘤微环境理论的提出使肿瘤细胞周围的非肿瘤细胞和基质以及肿瘤细胞与它们之间的相互作用逐渐受到重视。分析肿瘤的微环境,对探索肿瘤发生、进展、转移和治疗具有重要意义。现将Eph基因在肿瘤微环境调节中的作用和功能及与相关靶向治疗最新进展作一综述。     

局麻药长时间阻滞外周神经对神经源性疼痛发生过程中背根神经节内GAP-43表达的影响

用免疫组织化学方法观察了局麻药长时间阻滞外周神经对神经源性疼痛发生过程中生长相关蛋白(GAP-43)在背根神经节内表达的影响。实验选用SD大鼠35只,随机分成正常对照组、单纯坐骨神经横切组、坐骨神经横切前阻滞组、坐骨神经横切后阻滞组4大组,后三大组再按术后取材时间分为3、7d两个时间组。暴露大鼠右侧坐骨神经,于坐骨结节远端约1cm处横断坐骨神经。根据分组,阻滞组分别从横切前1h和横切后4h开始对神经横切的近心端进行长效局麻药阻滞,持续整个观察期。术后不同时间取与伤侧坐骨神经相连的背根神经节(DRG),应用免疫组织化学和图像分析的方法研究背根神经节中GAP-43的表达并进行定量分析。结果表明:术后3、7d,单纯坐骨神经横切组背根神经节内的GAP-43表达增高,而阻滞组内的GAP-43表达与正常组无差别。本研究结果提示坐骨神经横切前1h或之后4h开始局麻药阻滞神经均能抑制神经源性疼痛发生过程中GAP-43的高表达。     

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.     

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.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.