神经生长因子对谷氨酸介导的基底核神经元损伤的保护作用

本研究的目的是观察神经生长因子(nerve growth factor,NGF)对谷氨酸诱导的基底核神经元损伤的保护作用。取出生后1d乳鼠前脑基底核神经元培养,随机分为正常对照组、模型组(谷氨酸损伤组)和NGF保护组。用倒置相差显微镜进行活细胞观察,采用RT-PCR技术检测前脑基底核神经元的神经生长相关蛋白-43(growth associated protein-43,GAP-43)的表达。结果显示谷氨酸损伤组神经元胞体回缩,突起消失或断裂。NGF保护组的神经元绝大多数胞体饱满,突起明显,细胞间的网络联系仍清晰可见,接近于正常对照组;NGF保护组神经元内GAP-43mRNA表达比谷氨酸损伤组高,两者比较有统计学意义(P<0.05)。结果提示,NGF能保护基底核神经元免受谷氨酸兴奋性毒性的损伤。     

Neurturin和神经生长因子对体外培养的神经生长因子受体阳性神经元生长发育的影响

目的探讨Neurturin(NTN)、神经生长因子(NGF)对新生大鼠基底前脑胆碱能神经元生长发育的影响。方法用免疫组化结合图象分析技术观察Neurturin和NGF对培养的新生大鼠基底前脑NGF-R阳性神经元生长发育的作用。结果培养4d时,NTN组的NGF-R阳性神经元细胞数目、突起数、胞体面积和最长突起长度均与对照组有显著性差异(P<0.05),NGF+NTN组各项指标均优于NGF组或NTN组(P<0.05);培养8d时,NTN组除突起数目外其他各项指标均与对照组无显著性差异(P>0.05),NGF+NTN组各项指标仍然均优于NGF组或NTN组(P<0.05);NTN组NGF-R阳性神经元突起数多于NGF组(P<0.05)。结论Neurturin对体外培养的新生大鼠基底前脑NGF-R阳性神经元生长发育有营养作用,但作用时间较短,而且增加突起数的作用较NGF强;Neurturin和NGF的联合作用较单独使用Neurturin或NGF为好。     

神经节苷脂对损伤胎鼠背根神经节的保护作用

目的观察神经节苷脂(GM1)对受损伤胎鼠背根神经节(DRG)神经元形态改变的影响,探讨其可能的保护作用。方法选择胎龄为15d的SD大鼠为研究对象,获取DRG神经元并进行体外分散培养,培养48h后,随机分为对照组、谷氨酸损伤组和谷氨酸损伤+GM1保护组,继续培养12h。终止培养后,观察各组神经元的形态结构改变,用MTT法鉴定细胞的存活率。结果对照组DRG神经元细胞贴壁呈单层散在分布,少部分出现细胞聚集现象,突起较长且互相交织形成网状。谷氨酸损伤组DRG神经元细胞聚集现象明显,神经元突起变短、断裂甚至消失。谷氨酸损伤+GM1保护组DRG神经元细胞部分呈簇状聚集,部分呈单个散在分布,突起仍然相互交织。MTT结果显示谷氨酸损伤+GM1保护组细胞存活率高于谷氨酸损伤组。结论神经节苷脂可以影响损伤胎鼠DRG神经元的形态改变,对胎鼠背根神经节神经元具有一定的保护作用。     

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。     

BDNF、NGF对体外培养的胚胆碱能神经元生长发育的影响

本文用AChE组化方法，研究了BDNF、NGF对培养的胚鼠基底前脑胆碱能神经元的作用及BDNF和NGF的协同作用。结果表明BDNF和NGF都具有增加AChE阳性神经元数量的作用，二者的不同在于BDNF作用出现的时间较早、强度较小；而NGF作用出现的时间较迟但强度较大。并发现BDNF对体外培养的胚胆碱能神经元胞体早期的生长发育作用比较明显，而NGF的作用则不甚显著。BDNF对胚胆碱能神经元发出突起和突起的延伸作用较NGF强。BDNF和NGF的联合作用较单独使用BDNF或NGF为好。本文的结果提示在体外培养中两种营养因子联合应用较只用一种因子有益。     

中药神经再生素作用于背根神经节细胞过程中基因的表达变化

目的 探讨中药神经再生素(NRF)作用的分子生物学机制。方法 采用半定量PCR方法，观察和比较NRF组、NGF组和空白对照组中生长相关蛋白43(GAP-43)、低分子量神经丝蛋白(NF-L)、翻译延伸因子2A3-2(EF-Ts，RRAJ5161)和锌指样蛋白DDP2(F196315)基因水平的表达变化。结果 在NRF作用于离体培养的DRG细胞过程中，GAP-43、NF-L、2A3-2和DDP2基因表达在不同时间呈不同程度的上调。结论 NRF促神经生长的作用与NGF相似，可作用于蛋白翻译水平，促进神经细胞的生长；作用于细胞骨架蛋白水平，维持神经细胞的形态和神经细胞正常生理活动；作用于神经细胞特异的蛋白水平，具有维持细胞生存及促进神经细胞突起生长的作用；还可以作用于反式因子调控水平，影响相关基因的表达，促进神经元存活和神经突起延伸。     

Genistein对大鼠基底前脑胆碱能神经元发育影响的体外研究

本文旨在研究染料木素(genistein)对体外培养的基底前脑胆碱能神经元发育的影响。取孕18d胎鼠基底前脑神经元,体外有血清培养7d后,随机分为3组:无血清培养液组(control组),genistein+无血清培养液组(G组)、E2+无血清培养液组(E2组)。48h后用MTT法测定细胞活力和代谢状态;用AChE组化染色以及ChAT和MAP2免疫荧光双重染色,镜下计数AChE阳性和ChAT阳性神经元数,并对两种神经元的细胞面积、第一级突起数及最长突起长度进行检测。结果显示:MTT法所测的OD值、AChE阳性和ChAT阳性神经元数、细胞面积、第一级突起数及最长突起长度在G组与control组之间无明显差异,然而E2组中的上述数值均明显增加。本研究的结果提示:genistein对体外培养的基底前脑胆碱能神经元无类似雌激素样的神经保护和营养作用。     

银杏内酯对大鼠基底前脑神经元生长发育的影响

目的：观察银杏内酯对培养的胚胎大鼠基底前脑胆碱能神经元和NADPH-d阳性神经元生长发育的作用。方法：取E17Wistar大鼠胚胎基底前脑进行体外细胞培养，并给予银杏内酯。进行乙酰胆碱酯酶(acetyl cholinesterase，AChE)和NADPH-d酶组织化学染色，并在光镜下进行阳性细胞计数和显微测量胞体及突起发育状况。结果：给予银杏内酯处理后，培养物中乙酰胆碱酯酶阳性神经元和NADPH-d阳性神经元数量均较对照组显著增加，且神经元发育状况好于对照组，胞体大，突起多且长。结论：银杏内酯可以促进体外培养的基底前脑神经元中乙酰胆碱酯酶和NADPH-d的表达，并能促进表达这两种酶的神经元生长发育。     

牛膝多肽促大鼠海马神经元突起的生长

目的 研究牛膝多肽(ABPP)对体外培养的大鼠海马神经元突起生长的促进作用及其对生长相关蛋白-43 (GAP-43)和神经丝蛋白(NF-H)mRNA和蛋白表达的影响. 方法 以体外原代培养的胎鼠海马神经元为研究模型,通过四甲基偶氮唑盐(MTT)检测ABPP对海马神经元细胞活性的影响,通过免疫荧光细胞化学法,采用Image-Pro Express软件测量不同浓度ABPP(0.1mg/L、0.5mg/L、1.0mg/L)加药24h,对海马神经元神经突起生长的影响;通过实时荧光定量PCR法定量分析不同浓度ABPP(0.1mg/L、0.5mg/L、1.0mg/L)加药6h,对海马神经元GAP-43和NF-H基因表达的影响;采用免疫印迹法观察不同浓度ABPP(0.1mg/L、0.5mg/L、1.0mg/L)加药24h后,对海马神经元GAP-43 和NF-H蛋白表达的影响.同时应用胞外信号调节激酶(ERK)特异性拮抗剂PD98059与ABPP共培养海马神经元,分析ABPP对海马神经元的作用与ERK通路的关系. 结果 ABPP可有效地促进海马神经元突起的生长,加药后24h浓度为1.0mg/L,作用最强;实时荧光定量RT-PCR、免疫荧光细胞化学法和免疫印迹检测的结果表明,ABPP能增加体外培养的海马神经元GAP-43和NF-H的mRNA和蛋白表达,PD98059可抑制该过程. 结论 ABPP能促进体外培养的海马神经元神经突起的生长,其作用与上调GAP-43和NF-H 基因的表达相关,并可能与ERK通路有关.     

神经损伤与再生中神经元GAP-43mRNA表达的原位杂交研究

目的：研究周围神经损伤过程中生长相关蛋白（GAP-43）mRNA表达的变化规律。方法：建立大鼠坐骨神经中段钳夹损伤模型，用原位杂交技术对大鼠的腰髓和背根神经节的GAP-43mRNA表达进行观察。结果：大鼠坐骨神经损伤2d后，腰髓腹角运动神经元和背根节感觉神经元可检测到GAP-43mRNA杂交信号；术后4、7和14d明显增强；术后30d减弱，60d已恢复正常。结论：周围神经损伤诱导神经元胞体GAP-43mRNA表达显著增强，表明GAP-43在神经再生过程中起重要作用。     

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 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.