天麻素对脑缺血再灌注大鼠大脑皮质的影响及可能机制

目的 探讨天麻素对脑缺血再灌注大鼠大脑皮质的影响及抑制MAPK信号通路的可能机制.方法 24只SD大鼠随机分为正常对照组、脑缺血再灌注损伤组、天麻素组,每组8只.各组于再灌注72h后,通过神经功能评分及HE染色观察脑组织损伤情况;湿干重检测脑组织含水率;Western blot检测大鼠脑炎症相关指标IL-1β、NF-κB、TNF-α和通路蛋白p38MAPK、PI3K、p-Akt蛋白表达情况.结果 与对照组相比,损伤组神经功能评分值显著增高,大脑炎症反应加重,脑组织含水率均明显升高,IL-1β、NF-κB、TNF-α、p38 MAPK、PI3K和p-Akt蛋白表达水平上调(P＜0.05).与损伤组相比,天麻素处理后大鼠神经功能评分值显著降低,大脑炎症反应减轻,含水率显著降低,IL-1β、NF-κB、TNF-α、p38MAPK、PI3K和p-Akt蛋白表达水平下调(P＜0.05).结论 天麻素明显减轻脑缺血再灌注大鼠大脑皮质损伤程度,与调节MAPK信号通路减轻炎症反应相关.     

叔丁基对苯二酚对大鼠脑缺血再灌注损伤的保护作用及机制研究

目的:探讨叔丁基对苯二酚在大鼠脑缺血再灌注中的神经保护作用及可能机制。方法:将SD雄性大鼠随机分为假手术组、缺血再灌注组、缺血再灌注加溶剂治疗组、缺血再灌注加叔丁基对苯二酚治疗组,分别于再灌注后24 h对大鼠进行行为学评分,采用免疫印迹法检测叔丁基对苯二酚对Nrf2蛋白表达的影响,TUNEL法检测神经细胞的凋亡,酶联免疫吸附法检测炎性因子TNF-α和IL-1β的表达,同时检测超氧化物歧化酶和丙二醛的含量。结果:与假手术组相比,叔丁基对苯二酚可以显著上调Nrf2蛋白的表达(P0.05)和SOD的活性(P0.05),显著减少MDA的表达量(P0.05),抑制TNF-α和IL-1β的表达量(P0.05)。结论:叔丁基对苯二酚可以在大鼠脑缺血再灌注损伤中发挥显著地神经保护作用,其机制可能与上调Nrf2信号通路相关。     

三七总皂苷对大鼠局灶性脑缺血再灌注后IL-10及TNF-α表达的影响

目的观察三七总皂苷(PNS)对局灶性脑缺血再灌注后大鼠脑组织白细胞介素-10(IL-10)及肿瘤坏死因子(TNF-α)含量变化的影响,探讨PNS对脑缺血再灌注损伤的保护机制。方法将30只成年雄性SD大鼠随机分为假手术组、缺血再灌注组和PNS组,每组10只。缺血再灌注组采用线栓法制作大鼠大脑中动脉栓塞模型。各组分别于连续灌胃4周,观察3 d后处死取出脑脑织,采用ELISA法、免疫组织化学法和半定量RT-PCR法检测各组缺血区脑组织IL-10和TNF-α表达的变化,并进行大鼠行为能力测试评分。结果与假手术组和缺血再灌注组相比,PNS组术后脑组织中IL-10的表达水平明显增高,TNF-α的表达水平明显下调,PNS组行为能力较缺血再灌注组明显提高。结论三七总皂苷对脑缺血再灌注大鼠的保护作用,其作用机制可能与上调IL-10、降低TNF-α的表达有关。     

益气活血方和补肾生髓方对局灶性脑缺血再灌注大鼠缺血半暗带Notch-1和Jagged1表达的影响

目的:探讨益气活血方和补肾生髓方对脑缺血再灌注大鼠缺血半暗带Notch-1和Jagged1蛋白表达的影响。方法:线栓法制作局灶性脑缺血再灌注大鼠模型。随机分为假手术组、模型组、益气活血方防治组和补肾生髓方防治组。脑缺血2h,分别再灌注1周、2周、3周,采用免疫组化SABC法检测Notch-1和Jagged1蛋白表达的阳性总面积和平均吸光度。结果:在缺血侧额顶叶皮质缺血半暗带,再灌注各时点,模型组Notch-1和Jag-ged1蛋白表达的阳性总面积和平均吸光度显著高于假手术组(P0.05或P0.01),2种中药复方组再灌注各时点Notch-1和Jagged1蛋白表达的阳性总面积和(或)平均吸光度显著低于模型组(P0.05或P0.01)。结论:2种复方能通过降低缺血半暗带Notch-1、Jagged1蛋白表达,抑制Notch信号转导通路。     

参附注射液对脑缺血再灌注损伤大鼠Nrf2信号通路的影响

目的:探讨参附注射液治疗在大鼠脑缺血再灌注损伤中的保护作用及其对Nrf2信号通路的影响。方法:将68只雄性SD大鼠随机分为假手术组、脑缺血再灌注组、缺血再灌注损伤加参附注射液(8 mg/kg)治疗组,采用线栓法制作大鼠大脑中动脉缺血再灌注损伤模型,再灌注后24 h对大鼠进行行为学评分后处死并取脑,采用免疫印迹法检测参附注射液对Nrf2、HO-1、NQO1和cleaved-caspase-3蛋白表达的影响,尼氏法检测脑缺血再灌注损伤后的神经元损伤程度。结果:参附注射液治疗可以显著增加核内Nrf2蛋白的表达(P0.05),显著增加脑缺血再灌注损伤围脑组织中HO-1、NQO-1的表达量(P0.05),明显减少cleaved-caspase-3蛋白的表达和受损神经元的个数,显著改善神经功能评分(P0.05)。结论:参附注射液可以通过上调Nrf2信号通路从而在脑缺血再灌注损伤中发挥神经保护作用。     

NGF对局灶性脑缺血再灌注后大鼠皮质IL-1β表达的调控作用

目的:探讨外源性神经生长因子(NGF)对局灶性脑缺血再灌注大鼠皮质白细胞介素1β(IL-1β)表达的影响.方法:线栓法制作大鼠局灶性腩缺血再灌注模型,应用免疫组织化学显色、免疫印迹分析方法,结合图像分析技术检测大鼠缺血侧顶叶皮质 IL-1β蛋白表达变化.结果:假手术组大鼠顶叶皮质IL-1β没有表达,缺血再灌注组顶叶皮质IL-1β蛋白在各时间点明显表达,IL-1β蛋白阳性产物的光密度值高于假手术组;NGF组IL-1β蛋白阳性产物的光密度值低于缺血冉灌注组.结论:脑缺血再灌注损伤后诱导 IL-1β蛋白表达,NGF 明显降低缺血再灌注大鼠顶叶皮质IL-1β蛋白表达,这可能是 NGF发挥神经保护作用的分子机制之一.     

氟伐他汀预处理对脑缺血再灌注损伤大鼠脑组织TNF-α及IL-10表达的影响

目的观察氟伐他汀预处理对大鼠脑缺血再灌注损伤后脑组织肿瘤坏死因子(TNF-α)及白介素-10(IL-10)含量变化,探讨氟伐他汀的脑保护机制。方法实验大鼠随机分为4组:假手术组(Sham组)、脑缺血再灌注组(I/R组)、溶剂对照组(Vehicle组)和氟伐他汀预处理组(Flu组)。线栓法制作大脑中动脉脑缺血再灌注损伤模型,Flu组在模型制备前用氟伐他汀连续灌胃14 d。于缺血2 h再灌注24 h后断头取脑,采用ELISA法、免疫组织化学法和半定量RT-PCR法,检测大鼠脑组织中TNF-α及IL-10表达的变化。结果与Sham组比较,I/R组和Vehicle组脑组织中TNF-α和IL-10的表达明显上调(P<0.05)。与I/R组和Vehicle组比较,Flu组脑组织中TNF-α的表达明显下调,IL-10的表达显著上调(P<0.05)。结论氟伐他汀对脑缺血再灌注损伤的保护作用,其作用机制可能与上调IL-10,下调TNF-α的表达有关。     

腺苷预处理对大鼠脑缺血-再灌注损伤后脑内肿瘤坏死因子α和核因子κB表达的影响

目的 通过观察腺苷预处理对脑缺血-再灌注（IR）损伤后大鼠脑内肿瘤坏死因子α（TNF-α）和核因子κB（NF-κB）表达的影响,探讨腺苷对大鼠脑缺血-再灌注损伤的保护机制。方法 Sprague-Dawley大鼠60只随机分为假手术组（F组）、缺血-再灌注组（IR组）和腺苷预处理缺血-再灌注组（AP组）,每组动物按照缺血-再灌注后2 h、6 h、24 h及48 h这4个时间点随机分组,每组5只。采用大脑中动脉阻塞（MCAO）法制备大鼠大脑中动脉闭塞模型。通过对以上3组脑缺血 再灌注48 h后的大鼠脑组织行头颅MRI、HE染色以及对以上60只大鼠进行神经功能缺损评分,通过免疫组织化学方法观察大鼠脑组织中TNF-α和NF-κB的平均积分吸光度值进一步验证腺苷预处理对大鼠脑缺血-再灌注损伤的脑保护作用,探讨腺苷对大鼠脑缺血-再灌注损伤的保护机制。结果 F组大鼠术后无神经功能缺损体征,AP组和IR组大鼠术后均出现明显神经功能缺损体征。AP组和IR组大鼠神经功能缺损评分显著高于F组,AP组大鼠神经功能缺损评分显著低于IR组,差异均具有统计学意义（P＜0.05）。F组大鼠术后MRI中T1WI、T2WI均未见明显异常,AP组和IR组大鼠脑梗死体积显著高于F组,AP组大鼠脑梗死体积显著低于IR组,差异均具有统计学意义（P＜0.05）。AP组和IR组术后大鼠脑组织中TNF-α和NF-κB的表达显著高于F组,AP组术后大鼠脑组织中TNF-α和NF-κB的表达显著低于IR组,差异均具有统计学意义（P＜0.05）。结论 腺苷预处理可通过减少大鼠缺血-再灌注损伤后脑内TNF-α和NF-κB的表达而减轻神经细胞损伤,达到其脑保护作用。     

LRG介导内毒素预处理诱导的脑保护作用

目的:通过测定不同组大鼠脑缺血/再灌注损伤模型中脂多糖应答分子(LRG)表达水平的变化与炎症因子含量及大鼠脑梗死容积之间的关系,探讨内毒素重复预处理诱导脑保护效应的机制。方法:将66只SD大鼠随机分为假手术组、缺血对照组和内毒素预处理组,每组22只。采用戊巴比妥钠(40 mg/kg)腹腔注射麻醉大鼠,采用大鼠大脑中动脉栓塞模型(MCAO)制备大鼠大脑缺血/再灌注模型。采用免疫印迹法结合图像分析软件半定量计算LRG分子表达水平的变化,酶联免疫法测定脑组织TNF-α、IL-1β、IL-6、IL-10的含量,用氯化三苯四唑(TTC)染色法和计算机图像分析系统计算大鼠大脑梗死容积。结果:相比于缺血对照组,内毒素重复预处理组大鼠大脑LRG分子表达水平明显上调,TNF-α、IL-1β、IL-6含量明显降低,IL-10的含量明显增高,同时,内毒素预处理组较缺血对照组神经功能评分明显升高,脑梗死容积明显缩小。结论:内毒素重复预处理通过上调LRG分子表达,促进致炎-抑炎因子之间的动态平衡介导脑保护作用。     

脑缺血再灌注损伤后大鼠脑皮质TNF-α及其mRNA的时程变化

为了观察大鼠脑缺血再灌注损伤后大脑皮质TNF-α及其mRNA表达的时程变化,本实验采用大脑中动脉线栓法制作大鼠局灶性脑缺血再灌注模型,用免疫组化染色法检测不同再灌注时间大脑皮质TNF-α的蛋白表达水平,以及用逆转录-多聚酶链反应法(RT-PCR)检测脑皮质内TNF-αmRNA的表达情况。结果显示:脑缺血1h,再灌注12h后大脑皮质内TNF-α及其mR-NA的表达开始升高,24h后达到高峰,随后逐渐下降。两组大脑皮质TNF-α及其mRNA的表达水平差异有统计学意义(P<0.01),模型组的TNF-α及其mRNA的表达水平明显高于假手术组。本研究结果表明,TNF-α在大鼠脑缺血再灌注损伤的调节过程中发挥重要作用。     

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.     

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.     

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.     

The universal muscular chain reaction,muscle spasm,Torsions, ruptures and extravasations. Chameleons of pathology and some manifestations of simple muscular disorders

Most bodily functions require the coordinated actions of complementary and supplementary paired muscle groups. Where this essential muscular cooperation is lacking, hollow organs may burst and others become literally screwed up, giving rise to many similar spastic diseases such as Torticollis, Twisted ovarian cyst, Torsion of the Testis, Volvulus of the intestines, Varicose Veins, Megacolon, Aortamegaly, Scoliosis, Erb's Palsy, Peyronie's Disease, Main-en-Griffe, Undescended Foot (Pes Cavus), Talipes, Strabismus. Spasm is “panenepidemic” and unclassified examples of Torsion Dystonia and Dyskinesia really are as common as debt and taxes.     

Class II HLA in Georgia Caucasus Tbilisi Georgians and their Mediterranean ancestry: The Usko Mediterranean languages

Georgia (or Sakartvelo in its own language) is a South Caucasus Mts. country with its easternmost part is enigmatically named Iberia, like the Iberian Peninsula, which may refer to rivers “Kura” and “Ebro” or their valleys respectively. Most of their inhabitants speak Georgian which is included within Dene-Caucasian group and Usko-Mediterranean subgroup of languages. The latter includes Basque, Berber, ancient Iberian-Tartessian, Etruscan, Hittite, Minoan Lineal A and others. In the present paper, HLA class II -DRB1 and -DQB1 alleles has been studied and extended haplotypes calculated. Most frequent haplotypes are also of Mediterranean origin (i. e.: (A*02-B*51)-DRB1*11:01-DQB1*03:01, (A*02-B*51)-DRB1*13:01-DQB1*06:03, or (A*24-B*35)-DRB1*01:01-DQB1*05:01) and DA genetic distances show that closest world populations to Georgians are Mediterraneans. Georgians also show common extended haplotypes ((A*02-B*51)-DRB1*11:01-DQB1*03:01, (A*02-B*13)-DRB1*07:01-DQB1*02:01 and (A*03-B*35)-DRB1*11:01-DQB1*03:01) with Svan people, a secluded population in North Georgia mountains. We can conclude that Georgians belong to a very old Mediterranean substratum according to both linguistics (Usko Mediterranean languages) and HLA genetics.     

'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.