亚低温对脑出血大鼠血肿周围神经营养因子和血管新生的影响

目的:研究亚低温治疗对脑出血大鼠血肿周围脑组织中胶质细胞源性神经营养因子(GDNF)、血管内皮生长因子(VEGF)的影响。方法:雄性SD大鼠随机分为3组:假手术组(Sham组)、脑出血组(ICH组)和亚低温组。采用立体定向注射自体血复制脑出血大鼠模型,造模后15 min给予持续4 h的亚低温治疗。应用修正神经功能缺损评分(mNSS)对各组大鼠进行神经行为学评分;干湿重法检测各组大鼠脑组织含水量;Real-time PCR检测各组大鼠血肿周围脑组织内G-DNF、VEGF的mRNA表达水平;免疫印迹检测各组大鼠血肿周围脑组织内GDNF、VEGF的蛋白表达含量。结果:与假手术组相应时问点相比,脑出血组大鼠神经功能评分明显升高,脑组织含水量显著增多,血肿周围脑组织GDNF、VEGF的mRNA和蛋白表达明显上调;与脑出血组对应时间点相比,亚低温组神经功能评分明显降低,脑组织含水量显著减少,GDNF、VEGF的mRNA和蛋白表达显著上调。结论:亚低温治疗可增加脑出血大鼠脑内神经营养因子和血管生长因子的表达,促进神经修复和新生血管的形成,改善神经功能缺损和脑水肿。     

神经干细胞移植对脑外伤大鼠神经功能及脑组织Bcl-2与Bax表达的影响

目的探讨神经干细胞移植对脑外伤大鼠神经功能及脑组织Bcl-2与Bax表达的影响。方法108只SD成年大鼠随机分为假手术组(36只)、脑外伤组(36只)和神经干细胞移植组(36只)。采用改良的Feeney氏法制作大鼠脑外伤动物模型,损伤1周后在皮质处局部注射移植神经干细胞,分别在细胞移植后的第7天、14天和21天进行神经功能评分。免疫组织化学方法和Western blot方法检测各组大鼠脑组织Bcl-2与Bax蛋白的表达。结果脑外伤组大鼠出现明显的神经功能障碍,神经干细胞移植后第7、14和21天大鼠的神经功能评分明显降低(P0.05)。与假手术组相比,不同时间点脑外伤组大鼠脑组织Bcl-2蛋白的表达显著降低,Bax蛋白的表达显著升高(P0.05);与相同时间点的脑外伤组大鼠相比,神经干细胞移植组大鼠脑组织Bcl-2蛋白的表达显著升高,Bax蛋白的表达显著降低(P0.05)。结论神经干细胞移植促进脑外伤大鼠神经功能的恢复可能与上调Bcl-2蛋白和下调Bax蛋白表达相关。     

辛伐他汀对大鼠脑出血后TLR4介导的炎症损伤的干预作用

目的:探讨辛伐他汀(simvastatin,SIM)对大鼠脑出血后Toll样受体4(toll-like receptor 4,TLR4)介导的脑内炎症损伤的干预作用。方法:采用注入自体血方法复制脑出血(intralerebral hemorrhage,ICH)模型,随机分为假手术组、模型组和干预组。Garcia等方法进行神经功能障碍评分;干湿重法检测脑组织含水量;免疫组化SP法检测TLR4和IL-1β蛋白的表达;免疫印迹检测TLR4和IL-1β蛋白的表达水平。结果:与假手术组比较,ICH模型组脑组织含水量明显升高(P<0.05),TLR4和IL-1β蛋白的表达明显增加(P<0.05);辛伐他汀干预后,TLR4表达量下降,IL-1β蛋白的表达及脑含水量明显下调,与脑神经功能障碍评分上调相一致。结论:大鼠脑出血后引起出血周围脑组织炎症损伤,辛伐他汀可能通过减轻TLR4介导的炎症损伤,改善神经功能恢复,起到神经保护作用。     

辛伐他汀对实验性脑出血大鼠海马神经细胞自噬的影响

目的:观察辛伐他汀治疗对实验性脑出血大鼠海马区神经细胞自噬激活的调控作用。方法:将72只健康成年SD大鼠随机分三组(n=24):假手术组(Sham组)、脑出血组(ICH组)、辛伐他汀组(SIM组)。应用胶原酶诱导法建立脑出血大鼠模型,术后动物完全苏醒后给予辛伐他汀灌胃(3 mg/kg)。采用Garcia法对各组大鼠进行神经功能行为学评分,干湿比重法测定脑组织的水含量;免疫组织化染色学方法检测各组大鼠海马自噬微管相关蛋白1轻链3(Microtubule associated protein 1 light chain 3,LC3)的表达及分布;Westem blot法检测各组大鼠海马自噬基因Beclin-1表达。结果:与脑出血组相比,辛伐他汀治疗明显提高神经功能学评分(P0.05),减少脑组织含水量(P0.05),下调自噬相关蛋白LC3和Beclin-1的表达(P0.05),抑制海马神经元自噬的激活。结论:辛伐他汀能够降低脑出血大鼠海马区神经细胞自噬的激活,进而减轻脑水肿,促进神经功能的恢复。     

辛伐他汀预处理对脑缺血再灌注损伤Bcl-2和Bax表达的影响

目的:探讨辛伐他汀预处理对大鼠局灶性脑缺血再灌注损伤的保护作用及其机制。方法:随机将48只雄性SD大鼠分为4组:空白对照组、假手术组、缺血再灌注组和辛伐他汀预处理组。辛伐他汀预处理组在模型制备前用辛伐他汀20mg/kg连续灌胃10d,1次/d;假手术组及缺血再灌注组用相同体积的等渗盐水连续灌胃10d。以线栓法制作大鼠大脑中动脉缺血再灌注模型,于缺血2h再灌注24h后行5分制神经功能评分,并断头取脑分别测定脑梗死体积、凋亡细胞数及Bcl-2、Bax表达。结果:与空白对照组和假手术组比较,缺血再灌注组和辛伐他汀预处理组神经功能缺损较为严重,脑梗死体积增大,凋亡细胞数目增多,Bcl-2、Bax表达增加(均P0.01)。与缺血再灌注组相比,辛伐他汀预处理组神经功能有不同程度改善,脑梗死体积明显减小,凋亡细胞数及Bax表达降低,Bcl-2表达增高,比较差异均有统计学意义(P0.05,P0.01)。结论:辛伐他汀预处理对大鼠脑缺血再灌注有神经保护作用,其作用机制可能与辛伐他汀上调脑组织中Bcl-2、下调Bax表达,抑制细胞凋亡有关。     

白介素10对大鼠缺血梗死灶周围脑组织Bcl-2和Bax表达的作用研究

目的 探讨白介素10对大鼠脑缺血梗死灶周围Bcl-2和Bax表达的作用.方法 18只SD大鼠随机分为假手术组(Sham组)、脑缺血组(MCAO组)、脑缺血 溶剂对照组(Vehicle组)和脑缺血 IL-10干预组(IL-10组),免疫组化和RT-PCR法观察术后24h梗死灶周围脑组织Bcl-2和Bax表达水平的变化.结果与Sham相比,MCAO组梗死灶周围脑组织Bcl-2和Bax的表达显著上调,Bcl-2/Bax比值显著下降;与Vehicle组比,IL-10组Bcl-2的表达显著上调,Bax表达显著下调,Bcl-2/Bax比值显著增加.结论 IL-10与脑缺血梗死灶周围Bcl-2表达上调,Bax表达下调,Bcl-2/Bax比值增加有关,提示IL-10可能与抑制梗死灶周围脑组织神经细胞凋亡有关.     

黄连素对实验性脑出血大鼠脑水肿与神经功能的影响

目的:研究不同剂量黄连素对实验性脑出血(intracerebral hemorrhage,ICH)大鼠脑水肿与神经功能的影响,并初步探讨其作用机制.方法:实验共分为假手术组(Sham组),脑出血组(ICH组),药物低(Ber-10组)、中(Ber-20组)、高剂量组(Ber-40组),每组10只,除假手术组采用注射胶原酶的方法建立实验性脑出血大鼠模型,药物低、中、高剂量组分别给予黄连素10,20或40 mg/kg灌胃,每天1次,共给药28 d;对各组大鼠进行神经功能缺损评分,干湿重法测定脑含水量,Tunel法检测血肿周围脑组织细胞凋亡数,Western印迹法检测Cleaved cas-3,Cleaved cas-9,线粒体与胞浆中cyto-c的蛋白表达水平.结果:与脑出血组大鼠相比,采用黄连素给药后大鼠的神经功能缺损评分明显上升(P<0.05),脑含水量明显下降(P<0.05);血肿周围脑组织细胞凋亡数显著降低(P<0.05),Cleaved cas-3和Cleaved cas-9的蛋白表达,线粒体中cyto-c的蛋白表达明显上升(P<0.05)而胞浆中cyto-c的蛋白表达明显下降(P<0.05).结论:黄连素具有改善实验性脑出血大鼠神经功能缺损以及保护脑细胞的作用,可能与黄连素抑制线粒体凋亡通路有关.     

AQP4表达对脑出血大鼠模型脑水肿与相关凋亡蛋白影响

目的:研究AQP4表达对脑出血大鼠模型脑水肿与相关凋亡蛋白影响。方法:选取40只雄性SD大鼠随机分为对照组、模型组、AQP4siRNA组、空载质粒组,每组10只。模型组、空载质粒组、AQP4siRNA组均采用脑室内注射凝血酶Ⅶ注射建立大鼠尾壳核出血模型。AQP4siRNA组造模后将载有siRNA的质粒(10μl)注射于大鼠脑室内;空载质粒组造模后将不含AQP4siRNA片段的空质粒(10μl)注射于大鼠脑室内,用于排除质粒与手术对结果的干扰。术后1d和3d分别采用Longa分级法进行神经功能评分,术后3d后处死大鼠,采用干湿重法测量脑组织含水量,RT-PCR法检测血肿周围脑组织AQP4 mRNA表达,Western blotting法检测脑组织Caspase-3和Bcl-2蛋白表达水平。结果:与对照组比较,模型组神经功能评分明显增加,脑水肿明显,AQP4 mRNA表达明显增加,脑组织Caspase-3蛋白表达增加,Bcl-2蛋白表达降低(P0.05);与模型组比较,AQP4siRNA干预组可明显降低神经功能评分,减轻脑水肿(P0.05);同时AQP4 mRNA表达明显降低,脑组织Caspase-3蛋白表达明显降低,Bcl-2蛋白表达增加(P0.05)。结论:抑制AQP4基因表达能抑制脑水肿、减轻脑出血大鼠神经功能损伤,可能与减少Caspase-3和增加Bcl-2表达有关。     

远隔缺血后处理对脑缺血模型大鼠再灌注损伤炎性反应相关信号的影响

背景：虽然目前已有一些研究表明远隔缺血后处理可以发挥神经保护作用,但是具体的机制尚不明了。 目的：探讨远隔缺血后处理对大鼠局灶性脑缺血再灌注损伤的保护作用。 方法：应用线栓法制备大鼠大脑中动脉闭塞局灶性脑缺血再灌注模型,并进行远隔缺血后处理,同时设假手术组和缺血再灌注组作对照。于缺血再灌注24 h后进行神经功能评分,检测梗死体积及脑含水量;RT-PCR检测缺血周围区脑组织内白细胞介素1β、白细胞介素6、肿瘤坏死因子α和单核细胞趋化蛋白1 mRNA表达情况;Western blot检测Bcl-2和Bax蛋白表达情况。 结果与结论：与缺血再灌注组相比,远隔缺血后处理组神经功能评分有所降低,但差异无显著性意义,梗死范围和脑组织含水量显著降低(P < 0.05)。远隔缺血后处理组与缺血再灌注组相比,大鼠缺血周围区脑组织白细胞介素1β、白细胞介素6、肿瘤坏死因子α、单核细胞趋化蛋白1 mRNA和Bax蛋白表达降低(P < 0.05),Bcl-2蛋白表达升高(P < 0.05)。结果证实,远隔缺血后处理可以减轻大鼠局灶性脑缺血再灌注产生的损伤,其机制可能与减轻炎性反应有关。  中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程     

黄芪注射液对大鼠脑出血后神经的保护作用

目的探讨黄芪对大鼠脑出血灶周围凋亡神经元的保护作用。方法用Ⅶ型胶原酶脑内注入法制作大鼠脑出血模型,对各组大鼠进行神经功能评分,用免疫组织化学法检测Bcl-2和Bax蛋白表达,透射电镜观察大鼠脑出血灶周围神经元的超微结构,并对神经突触进行定量分析。结果与模型组比较,各治疗组Bcl-2的阳性表达均明显升高(P0.01);各治疗组Bax的阳性表达均明显降低(P0.01);Bcl-2/Bax蛋白比值增高。与术后0h、6h治疗组相比,术后24h治疗组Bcl-2和Bax的阳性表达有显著性差异(P0.05)。术后0h治疗组与术后6h治疗组相比,Bcl-2阳性表达无显著性差异(P0.05)。神经元超微结构观察,正常组神经元超微结构正常;模型组神经元超微结构严重破坏;各黄芪治疗组神经元超微结构与模型组相比明显好转。其中出血后0h、6h治疗组的神经元超微结构好于出血后24h治疗组。神经突触定量分析显示,与模型组相比,各治疗组神经突触的数量明显增多(P0.01);突触间隙明显变窄(P0.01);突触后致密区明显增厚(P0.01)。术后24h治疗组与术后0h、6h治疗组相比,各定量参数均有显著性差异(P0.05)。结论黄芪能保护大鼠脑出血灶周围的神经元,增强神经突触功能,减少神经元凋亡,并且早期用药效果较好。     

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.