急性脱髓鞘对小鼠海马和大脑皮层中Nkx2.2的影响

目的:探讨双环己酮草酰二腙(CPZ)诱导急性脱髓鞘后小鼠认知功能的改变,以及海马和大脑皮层中Nkx2.2同源域转录因子(Nkx2.2)的表达变化,并探讨两种变化之间的关系。方法:0.2%CPZ喂养ICR小鼠6周建立急性脱髓鞘模型。通过Morris水迷宫(MWM)和悬尾实验(TST)检测CPZ介导的急性脱髓鞘后小鼠行为学的改变; Western Blot检测脱髓鞘后小鼠海马和大脑皮层中髓鞘碱性蛋白(MBP)和Nkx2.2蛋白的表达;免疫组织化学检测CPZ脱髓鞘后小鼠海马和大脑皮层中Nkx2.2的表达变化。结果:MWM和TST结果显示:与对照组小鼠相比,CPZ组小鼠MWM实验中的逃避潜伏期时间增加,穿越平台次数减少,TST的悬尾不动时间增加(P 0.05); Western Blot结果显示:CPZ组小鼠海马和大脑皮层中的MBP蛋白含量均明显低于对照组(P 0.05),Nkx2.2蛋白含量均明显高于对照组(P 0.05);免疫组织化学结果显示:CPZ组小鼠海马和大脑皮层中Nkx2.2的免疫阳性表达均高于对照组(P 0.05)。结论:CPZ介导急性脱髓鞘后小鼠的认知功能下降,海马和大脑皮层中Nkx2.2的表达增加,提示Nkx2.2在脱髓鞘模型中的异常增加可能是导致小鼠认知功能下降的原因之一。     

肌苷对CPZ介导的急性脱髓鞘小鼠行为学及皮质髓鞘的影响

目的:探讨肌苷对双环己酮草酰二腙(cuprizone,CPZ)介导的急性脱髓鞘小鼠行为学及皮质髓鞘的影响。方法:在普通饲料中掺入0.2%CPZ,饲养小鼠6 w,同时联合腹腔注射肌苷,制备脱髓鞘治疗模型,利用体重测量、Morris水迷宫、悬尾实验、透射电镜等技术,观察肌苷对脱髓鞘小鼠治疗后行为学及皮质髓鞘的影响。结果:(1)体重变化:与生理盐水对照组比较,CPZ损伤组、肌苷治疗组小鼠体重从第6 d开始均明显降低(P0.05);(2)行为学:Morris水迷宫空间定位实验中,与CPZ损伤组比较,肌苷治疗组平台象限停留时间明显增加(P0.05);悬尾实验中,小鼠6 min不动时间均无明显差异(P0.05);(3)皮质髓鞘电镜观察显示,肌苷治疗组小鼠的皮质髓鞘病理改变程度降低,有新生髓鞘。结论:通过含0.2%CPZ饲料饲养联合腹腔注射肌苷制备脱髓鞘治疗小鼠,可使小鼠的学习、记忆功能明显改善,皮质髓鞘有明显的修复与再生,揭示肌苷对伴有学习记忆功能障碍的脱髓鞘小鼠有髓鞘保护作用。     

Nlrp3敲减对cuprizone诱导的脱髓鞘小鼠焦虑症状的影响

目的:探讨NOD样受体家族pyrin域3(Nod like receptor family pyrin domain containing 3,Nlrp3)对脱髓鞘损伤致小鼠焦虑情绪的影响。方法:雄性C57BL/6小鼠分为对照组、双环己酮草酰二腙(CPZ)模型组、CPZ+Nlrp3 shiNA组和Nlrp3 shRNA组;将对照或Nlrp3 shRNA慢病毒分别立体定位注射至小鼠胼胝体。1周后,小鼠喂食CPZ并测量喂食期间小鼠体质量;开场实验检测小鼠活动区域与距离,LFB-PAS染色观察脱髓鞘程度。结果:CPZ喂食2周内,小鼠平均体质量急剧下降;到5周时,CPZ模型组平均体质量显著低于对照组与CPZ+Nlrp3 shRNA组。行为学检测显示,对照组和CPZ+Nlrp3 shRNA组小鼠后肢站立次数较CPZ模型组明显减少,而中央区域活动距离与活动总距离的比值CPZ模型组显著增加。LFBPAS染色显示CPZ模型组胼胝体膝部髓鞘几乎完全缺失,其髓鞘化评分显著低于正常和CPZ+Nlpr3shRNA组。结论:Nlrp3敲减可缓解CPZ诱导的小鼠髓鞘脱失,并改善其焦虑症状。     

帕金森病MPTP模型小鼠海马内星形胶质细胞活化和p-CREB的表达下降

目的:通过检测帕金森病(Parkinson's disease,PD)MPTP模型小鼠海马内GFAP和p-CREB的表达,探讨PD认知障碍发生的原因。方法:雄性C57BL/6小鼠随机分为对照组和PD模型组,每组12只。PD模型组经腹腔注射MPTP和丙磺舒,3.5 d注射1次,共10次,持续5周,对照组小鼠注射等量生理盐水。在第6周,旷场试验和爬杆试验评价运动功能。免疫组化检测黑质、纹状体TH的表达及海马GFAP和p-CREB的表达;Western Blot检测p-CREB的表达。结果:旷场实验和爬杆实验结果显示PD模型组小鼠运动能力明显低于对照组(P0.001)。免疫组化显示PD模型组黑质TH阳性神经元和纹状体TH阳性神经纤维均明显低于对照组(P0.001),海马的GFAP阳性细胞的面积明显多于对照组。而Western Blot和免疫组化染色均显示PD模型组海马p-CREB的表达明显少于对照组(P0.001)。结论:慢性PD模型小鼠海马内星形胶质细胞激活和p-CREB表达降低,可能参与了PD认知损害的病理过程。     

模型小鼠卵巢切除对大脑Basigin表达的影响

目的:用切除雌性小鼠卵巢研究Basigin在脑皮质和海马中表达,以确定雌激素调控Basigin状况。方法:选择雌性昆明小鼠28只,体重(26±2)g,3月龄。小鼠随机分为切除卵巢组(n=14)和假手术组(n=14)。小鼠切除卵巢组经背侧行卵巢切除术,假手术组只切除卵巢周围相应大小的脂肪组织,而不切除卵巢。2个月后将小鼠处死取出脑组织,选择脑皮质区和海马区分别用免疫组织化学和Western Blot技术测定Basigin表达水平。结果经统计学分析。结果:小鼠脑皮质区和海马区均可见细胞质和胞膜中Basigin阳性表达。免疫组化表明,与假手术小鼠比较,切除卵巢小鼠皮质区和海马CA1、CA3区Basigin表达明显下调(P0.05~0.01),但海马CA4变化不明显。Western Blot检测结果显示,切除卵巢小鼠脑皮质区和海马区Basigin表达明显低于假手术小鼠(P0.05)。结论:卵巢切除小鼠脑皮质区和海马区Basigin表达明显下调,表明雌激素在调节脑皮质区及海马区Basigin表达具有重要作用。     

神经调节蛋白1及其受体ErbB4在精神分裂症小鼠前额叶皮质中的表达变化

目的:探讨精神分裂症神经调节蛋白1(NRG-1)及其受体(ErbB4)与少突胶质细胞脱髓鞘和髓鞘再生之间的关系。方法:采用双环己酮草酰二腙(CPZ)诱导的少突胶质细胞脱髓鞘制备精神分裂症小鼠动物模型。动物分为6组,对照组(食物中未添加CPZ),CPZ喂养2周、3周、4周组,第4周后停止给予CPZ,继续正常喂养1周即5周组;停止给予CPZ后,继续正常喂养2周即6周组。通过实时定量PCR及免疫印迹检测,观察NRG-1和ErbB4在小鼠大脑额叶脱髓鞘和髓鞘再生过程中的表达变化。结果:NRG-1 mRNA和蛋白的表达在第2周、第3周和第4周CPZ组较对照组、第5周和第6周明显降低。ErbB4 mRNA和蛋白的表达在各组与对照组比较均增强。ErbB4在第5周和第6周mRNA和蛋白的表达明显高于第2周、第3周和第4周CPZ组。结论:在小鼠精神分裂症形成过程中,NRG-1和ErbB4的表达变化与少突胶质细胞的脱髓鞘和髓鞘再生有关系。     

低氧预适应对小鼠海马神经元中TSC1表达的影响

目的:在体视显微镜下分割ICR小鼠海马CA1区和CA3区,研究结节性硬化症因子1(TSC1)在小鼠海马低氧中的神经保护作用。方法:ICR小鼠分为对照组(control)、低氧对照组(hypoxia)及低氧预适应组(HPC),在体视显微镜下观察海马形态并分割CA1区和CA3区;采用real time RT-PCR和Western Blot的方法分别检测小鼠海马组织TSC1 mRNA和蛋白的表达;采用免疫荧光检测小鼠海马组织TSC1荧光强度。结果:大脑冠状切片清晰显示出海马CA1区、CA3区和DG区;CA1区TSC1 mRNA在低氧组降低而在低氧预适应组增高;Western Blot和组织免疫荧光显示:与对照组相比,低氧预适应组CA1区TSC1表达增加;而CA3区TSC1在低氧组和低氧预适应组均增加。结论:TSC1的差异性表达可能提示TSC1可能参与了低氧预适应对低氧敏感的CA1区神经细胞的保护。     

狼疮性肾炎肾小球中nestin和活化型Caspase 3的表达及意义

目的探讨狼疮鼠肾小球中nestin和足细胞中活化型Caspase 3的表达变化及其与小鼠蛋白尿的关系。方法选取30周龄雌性MRL/MPJ小鼠和MRL/faslpr小鼠为研究对象,分别设为对照组和狼疮性肾炎模型组。血、尿生化检测小鼠肾功能改变;采用免疫组化和Western blot技术检测小鼠肾小球中活化型Caspase 3蛋白的表达;免疫组化法检测小鼠肾小球中nestin蛋白的表达;免疫荧光双染技术检测肾小球中活化型Caspase 3与nestin共表达。结果 (1)与正常对照组相比,狼疮组小鼠24 h尿蛋白(Upro)明显升高(P0.05),而尿素氮(BUN)和血清肌酐(Scr)在两组之间差异无显著性。(2)透射电镜结构显示,狼疮性肾炎模型组小鼠肾小球上皮细胞出现足突融合,基膜出现明显的不规则增厚。(3)与正常对照组相比,活化型Caspase 3在狼疮性肾炎模型组小鼠肾小球中表达增高;(4)免疫组化结果显示,nestin阳性信号主要定位于细胞质和细胞核,定量分析结果显示,与正常对照组相比,狼疮性肾炎模型组肾小球中nestin蛋白表达上调。(5)免疫荧光双染检测结果显示,正常对照组中仅见nestin蛋白表达,未见明显的nestin和Caspase 3共定位表达,在狼疮性肾炎模型组小鼠肾小球中出现了nestin蛋白和Caspase 3蛋白明显共定位。(6)相关性分析:狼疮性肾炎模型组小鼠肾小球中活化型Caspase 3蛋白的表达量与24 h Upro呈明显正相关(r=0.827,P=0.006)。结论足细胞骨架结构的改变和(或)足细胞凋亡可能在狼疮性肾炎发病过程中发挥重要的作用。     

银杏提取物对实验性自身免疫性脑脊髓炎小鼠的影响

目的:探讨银杏提取物(GBE)对实验性自身免疫性脑脊髓炎(EAE)小鼠炎症脱髓鞘病变的影响。方法:应用髓鞘少突胶质细胞糖蛋白33-55(MOG33-55)配以完全弗氏佐剂(CFA)免疫小鼠,诱发EAE模型。将小鼠分为CFA对照组、EAE模型组和GBE治疗组(每日腹腔注射GBE70mg/kg)。通过神经功能评分、行为学实验以及免疫荧光染色,观察GBE对EAE小鼠的影响。结果:GBE组小鼠各时间段神经功能评分均低于EAE组(P0.05),行为学检测显示发病高峰期falling latency时间较EAE组延长10s;GBE组较EAE组视神经髓鞘碱性蛋白(MBP)表达水平增高,可见MBP阳性髓鞘结构包绕轴突;海马伞矢状切片免疫荧光染色证实GBE组CD11b阳性小胶质细胞较EAE组明显减少,但是GFAP阳性星形胶质细胞数量与EAE组无明显差别。结论:GBE可能通过抑制小胶质细胞激活从而延缓EAE小鼠脱髓鞘进程,提示GBE对多发性硬化具有一定的治疗作用。     

Cuprizone诱导髓鞘损伤再生模型中Sip1的表达变化及其意义

目的:研究Smad相互作用蛋白1(Sip1)在cuprizone诱导髓鞘损伤再生模型中的表达变化及其意义。方法:通过在饲料中掺入cuprizone喂食C57BL/6小鼠建立髓鞘损伤模型,利用黑金(black gold,BG)染色方法及Western Blot方法,检测髓鞘损伤模型是否建立成功;利用Western Blot方法检测在髓鞘损伤及再生过程中Sip1的表达情况。结果:BG染色方法检测到喂药6周后模型组小鼠与对照组相比,着色显著降低,Western Blot方法检测到模型组小鼠MOG(myelin oligodendrocyte glycoprotein)蛋白表达水平显著降低,GFAP(glial fibrillary acidic protein)蛋白表达水平显著增高,证明髓鞘损伤模型建立成功。在模型建立成功后,停止喂药,改用正常饲料喂养4周后,通过Western Blot检测到模型组小鼠MOG蛋白水平显著恢复,证明该阶段髓鞘已再生。利用Western Blot方法检测髓鞘损伤模型建立阶段及髓鞘再生阶段的Sip1蛋白水平,结果显示与对照组小鼠相比,在髓鞘损伤阶段,模型组小鼠的Sip1蛋白水平显著增加,在髓鞘再生阶段Sip1蛋白水平同样显著增加。结论:Sip1在髓鞘损伤再生过程中具有重要作用,本研究为Sip1作为治疗髓鞘相关疾病的靶点提供了理论依据。     

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 case for allele‐specific recognition by the TCR

There is a sharp difference in how one views TCR structure–function–behaviour dependent on whether its recognition of major histocompatibility complex‐encoded restriction elements (R) is germline selected or somatically generated. The generally accepted or Standard model is built on the assumption that recognition of R is by the V regions of the αβ TCR, which is not driven by allele specificity, whereas the competing model posits that recognition of R is allele‐specific. The establishing of allele‐specific recognition of R by the TCR would rule out the Standard model and clear the road to a consideration of a competing construct, the Tritope model. Here, the case for allele‐specific recognition (germline selected) is detailed making it obvious that the Standard model is untenable.     

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.