臂丛根性撕脱伤激活脊髓表达磷酸化CaMKⅡ的时程及定位研究

目的研究大鼠臂丛根性撕脱伤后磷酸化CaMKⅡ的时间和空间定位。方法将SD大鼠（6-8周龄）在手术显微镜下做臂丛根性撕脱伤手术后,应用Western blot方法测定相应脊髓节段（C7、C8）磷酸化CaMKⅡ在1、2、3d时间点及正常大鼠的表达水平。同时应用免疫荧光染色方法检测目标蛋白在1d时间点大鼠脊髓节段的表达水平及空间定位。结果正常大鼠脊髓相应节段检测不到磷酸化CaMKⅡ,在臂丛根性撕脱伤后1～2d对应脊髓节段的磷酸化CaMKⅡ表达明显升高,撕脱伤后3d明显下降并接近正常水平;免疫荧光结果显示：臂丛根性撕脱伤后1d,损伤侧脊髓前角运动神经元和中间神经元明显表达磷酸化CaMKⅡ,对侧脊髓前角运动神经元和中间神经元也有表达。但强度较撕脱侧明显减弱。结论臂丛根性撕脱伤诱导磷酸化CaMKⅡ在损伤节段1d时高表达,然而在3d时逐步下降到正常水平。     

臂丛损伤脊髓运动神经元与神经根GAP-43 mRNA表达

目的：探讨臂丛根性撕脱伤后脊髓腹角运动神经元胞体及其神经根GAP-43 mRNA的表达变化及其影响因素,为臂丛损伤的修复治疗提供理论依据.方法：本实验创立三种臂丛根性撕脱伤模型：C7前根撕脱（Ⅰ组）;C7前根撕脱＋切断同侧C5～T1后根（Ⅱ组）;C7前根撕脱＋C5和C6之间作同侧脊髓半横断（Ⅲ组）.术后2周按CBS评分标准检查动物神经缺失症状,用SYBR Green荧光定量RT-PCR方法检测脊髓腹角运动神经元胞体及其神经根GAP-43 mRNA的表达改变.结果：根据CBS评分标准,对照组计为0分,Ⅰ组计分较低、Ⅲ组计分最高.对照组C7神经元胞体和C7神经根中GAP-43 mRNA表达量相近,但三种损伤组术后2周神经元胞体内GAP-43 mRNA表达均上调,而神经根内表达却下调.结论：（1）臂丛根性撕脱伤后脊髓腹角运动神经元胞体GAP-43 mRNA表达受突触前机制的调控;（2）臂丛损伤2周时神经元胞体内GAP-43 mRNA表达呈现高峰期,此时进行神经移位术将显著提高神经修复的效果.     

臂丛根性撕脱伤激活脊髓表达磷酸化CaMKⅡ的时程及定位研究

目的研究大鼠臂丛根性撕脱伤后磷酸化CaMKⅡ的时间和空间定位。方法将SD大鼠(6～8周龄)在手术显微镜下做臂丛根性撕脱伤手术后,应用Western blot方法测定相应脊髓节段(C7、C8)磷酸化CaMKⅡ在1、2、3 d时间点及正常大鼠的表达水平。同时应用免疫荧光染色方法检测目标蛋白在1 d时间点大鼠脊髓节段的表达水平及空间定位。结果正常大鼠脊髓相应节段检测不到磷酸化CaMKⅡ,在臂丛根性撕脱伤后1～2 d对应脊髓节段的磷酸化CaMKⅡ表达明显升高,撕脱伤后3d明显下降并接近正常水平;免疫荧光结果显示:臂丛根性撕脱伤后1 d,损伤侧脊髓前角运动神经元和中间神经元明显表达磷酸化CaMKⅡ,对侧脊髓前角运动神经元和中间神经元也有表达,但强度较撕脱侧明显减弱。结论臂丛根性撕脱伤诱导磷酸化CaMKⅡ在损伤节段1 d时高表达,然而在3 d时逐步下降到正常水平。     

臂丛根性撕脱伤后神经根回植术的大鼠动物模型

目的：建立合理的臂丛根性撕脱伤后神经根回植术的大鼠动物模型。方法：在手术显微镜下，采用前入路，将C6神经根从脊髓上撕脱，咬除同侧C5椎体下外部分，显露脊髓；切断肌皮神经，切取长约30mm尺神经桥接肌皮神经与脊髓间的缺损，并将神经近端植入脊髓。术后观察手术侧前后肢的一般情况；6个月后，观察神经的解剖与组织学的连续性。结果：大鼠存活良好，手术侧前肢无坏死、溃疡、脱落，后肢无瘫痪；从脊髓到肱二头肌，神经的连续性完整；组织学检查见桥接神经段内有神经纤维再生。结论：该模型显露脊髓和切取桥接用神经方便，再植位置准确，便于直接观察神经根再植后神经再生及功能恢复情况，无明显的脊髓损伤并发症，较好模拟了臂丛根性撕脱伤后神经前根的回植。     

RNA干扰技术抑制nNOS基因对根性撕脱伤脊髓运动神经元的影响

目的研究神经元型一氧化氮合酶(neuronal nitric oxide synthase,nNOS)基因在臂丛神经根撕脱伤中的作用地位。方法设计、筛选并构建针对nNOS基因的siRNA表达载体,建立C5～T1神经根撕脱伤SD雄性大鼠模型,于撕脱后14d脊髓鞘内给予nNOS的siRNA干预,3d后用NADPH-d酶组化反应结合中性红染色评估撕脱伤脊髓运动神经元的nNOS基因表达水平和创伤神经元的存活率。结果臂丛C5～T1根性撕脱伤17d后,C7节段损伤侧前角运动神经元的存活率在siRNA组、siRNA序列对照组和生理盐水组分别为(80.51±9.16)%、(88.26±2.95)%和(89.13±3.47)%;C7节段损伤侧前角运动神经元的nNOS阳性率为(18.10±6.63)%、(42.21±2.29)%和(39.46±2.41)%。nNOS的siRNA能显著减低撕脱伤诱导的nNOS蛋白在前角运动神经元内的表达水平。虽然与对照组相比,nNOS-siRNA组撕脱伤后运动神经元存活率有下降趋势,nNOS的siRNA但并未对撕脱伤导致的运功神经元的死亡造成显著影响。结论鞘内应用siRNA技术能下调撕脱伤后大鼠脊髓运动神经元内nNOS蛋白的表达,nNOS基因有可能发挥维持撕脱伤后运动神经元存活的作用。     

臂丛损伤后脊髓前角运动神经元超微结构改变

目的探讨大鼠臂丛损伤导致脊髓前角运动神经元死亡的机制。方法成年雄性SD大鼠24只,其中对照组6只,损伤组18只。建立3种臂丛损伤模型:右C7前根撕脱(A组);右C7前根撕脱+同侧C5～T1后根离断(B组);右C7前根撕脱+右C5与C6之间脊髓半横断(C组)。术后14d取C7节段脊髓,采用尼氏染色方法和透射电镜技术,观察脊髓前角运动神经元的存活率及其超微结构改变。结果术后2周A组脊髓前角运动神经元的存活率最高,B组居中,C组最低。3个臂丛损伤组C7前角均可见凋亡特征性改变:运动神经元内核染色质聚集靠边,核固缩、碎裂、核膜皱褶并内陷,并有染色质团块形成的凋亡小体。细胞体积缩小,胞浆内细胞器密集,线粒体轻度肿胀,核周粗面内质网减少,游离核糖体增多,胞浆内可见较多的空泡。神经元胞体周围的有髓神经纤维和无髓神经纤维呈轻度肿胀,髓鞘的板层结构消失。结论臂丛损伤诱导脊髓运动神经元死亡途径中存在凋亡和坏死两种机制,运动神经元可形成凋亡小体。     

葛根素对臂丛根性撕脱后脊髓MMP-9蛋白表达的影响

目的　探讨大鼠臂丛根性撕脱后脊髓基质金属蛋白酶-9（MMP-9）蛋白的表达变化以及葛根素对其表达的影响。 方法　成年雄性SD大鼠174只，随机分为正常组、模型组、葛根素低、中、高处理组。模型组和葛根素处理组行右侧C5~C7脊神经前根撕脱术，术后腹腔注射给药。Western Blot法检测C5~C7节段脊髓MMP-9蛋白的表达。 结果　与正常组比较，臂丛神经根性撕脱后1 d时MMP-9蛋白表达达高峰，3 d降至接近正常，1周后下降至较低水平。葛根素处理组脊髓组织MMP-9蛋白表达在1 d时比模型组显著降低。 结论　MMP-9可能参与臂丛根性撕脱后早期损伤反应，且葛根素可能有助于减轻该种损伤。     

臂丛损伤再生中GAP-43 mRNA及其蛋白的表达

目的：分析臂丛损伤后脊髓前角运动神经元表达GAP-43 mRNA及其蛋白的变化规律，探讨神经损伤再生的机制。方法：建立3种臂丛损伤模型：右C₇前根撕脱（A组）；右C₇前根撕脱＋同侧C5-T₁后根断离（B组）；右C₇前根撕脱+右C₅C₆间脊髓半横断（C组）。用荧光定量RT-PCR方法检测术后14 d时 C₇前角GAP-43 mRNA的表达量。用免疫组化方法检测术后1、 3、 7、14 d脊髓前角GAP-43免疫阳性运动神经元的表达。结果：对照组C₇前角GAP-43 mRNA呈低表达，损伤组GAP-43 mRNA表达显著上调。损伤组术后1 d、3 d时均未见C₇前角 GAP-43免疫阳性神经元，术后7 d各损伤组GAP-43免疫阳性神经元开始出现，14 d时免疫阳性神经元数目达到高峰。3组间比较，C组表达量最高，B组最低，A组居中。结论：臂丛损伤诱导运动神经元GAP-43 mRNA及其蛋白表达上调，GAP-43合成增加是神经元蛋白重组所致，与轴索再生和神经功能重建有关。     

电针疗法对臂丛根性撕脱伤脊髓后角及中央管nNOS蛋白表达的影响

目的采用电针疗法干预大鼠臂丛神经撕脱伤模型,探索电针疗法对臂丛根性撕脱伤脊髓后角及中央管n NOS蛋白表达的影响。方法健康、雌性成年Sprague-Dawley(SD)大鼠共40只,行臂丛神经根性撕脱手术,随机分为撕脱伤组(AV组)和撕脱伤加电针治疗组(AV+EA组),AV+EA组动物隔日接受大椎(DU4)和手三里(LI10)电针治疗直至处死,每次治疗的输出脉冲波形为非对称双向疏密波,以20 Hz频率不间断治疗15 min。动物存活1周、2周、3周、6周处死,选取C7节段脊髓,行NADPH-d酶组织化学染色和中性红复染。结果脊髓后角,在AV组n NOS的微量表达;在AV+EA组2~3周n NOS在健侧的表达比伤侧增多,至6周脊髓后角n NOS阳性神经元表达AV组损伤侧;AV+EA组n NOS阳性神经元的表达与同时期的AV组。结论电针疗法导致脊髓后角及中央管n NOS阳性神经元的时空表达特异性。     

小动物PET-CT在大鼠臂丛根性撕脱脊髓损伤的应用研究

目的探究使用小动物正电子放射断层扫描(PET)/计算机断层成像(CT)技术诊断大鼠神经根撕脱导致的脊髓损伤的程度。方法对正常SD大鼠,通过尾静脉或腹腔注射不同剂量18F-FDG,筛选最优注射方式及剂量;随后,建立SD大鼠右侧全臂丛根性撕脱模型(BPRA组)并以假手术大鼠为对照组,对术后2周大鼠进行18F-FDG-micro-PET-CT成像检测,计算脊髓节段每克组织放射性占注入量的百分比(%ID/g),比较BPRA和假手术组大鼠C5~T1脊髓组织18F-FDG摄取差异。结果尾静脉注射1μCi/g的示踪剂18F-FDG,40 min后进行micro-PET-CT扫描其效果最佳;假手术组大鼠下颈段脊髓节段18F-FDG摄取均匀,而右全臂丛根性撕脱伤术后2周,BPRA模型组颈段脊髓18F-FDG摄取高亮范围增大,脊髓C5~C8、T1节段18F-FDG摄取率(%ID/g)为0.69±0.04与假手术组(0.60±0.02)比显著增加(P0.05)。结论尾静脉注射18F-FDG 1μCi/g联合micro-PET-CT成像技术能够监测臂丛撕脱伤后在体的脊髓组织的病理反应所引起的局部神经元和胶质细胞的代谢变化,为该病的基础研究提供工具及临床病程诊断提供了新思路。     

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.