天麻素对氧糖剥夺诱导的小胶质细胞Notch信号通路的影响

目的:研究天麻素(GAS)对氧糖剥夺(OGD)激活的BV2细胞Notch通路表达的影响.方法:BV2细胞分成4组,分别为对照组(control)、氧糖剥夺组(OGD)、OGD联合0.17 mmol/L GAS处理组(OGD+GAS-L),和OGD联合0.34 mmol/L GAS处理组(OGD+GAS-H).免疫荧光双...     

mGluR4激动剂对氧糖剥夺SH-SY5Y细胞损伤的保护作用研究

目的:研究m GluR4在SH-SY5Y细胞氧糖剥夺模型中的预防作用,并探讨其相关机制。方法:维甲酸(RA)诱导体外培养SH-SY5Y并构建氧糖剥夺模型,将细胞分为对照组、氧糖剥夺组(OGD)、OGD+m GluR4激动剂VU0155041处理组、OGD+VU0155041+SB202190处理组。CCK8检测细胞存活率; Annexin V-FITC/PI双染法检测细胞凋亡,Western Blot检测蛋白表达水平;流式细胞仪检测细胞增殖。结果:RA诱导的SH-SY5Y细胞中广泛表达m GluR4,而m GluR4在小胶质瘤细胞中不表达; 10μmol/L的m GluR4激动剂VU0155041可显著预防OGD处理的SH-SY5Y细胞活力; m GluR4激动剂VU0155041可显著预防OGD引起的SH-SY5Y细胞凋亡,并且m GluR4激动剂VU0155041对细胞的保护作用与增殖无关; OGD可促进p-p38 MAPK通路的激活,并上调Cytc、active caspase 3水平;而m GluR4激动剂VU0155041可下调p-p38 MAPK水平,下调Cytc、active caspase 3水平,抑制细胞凋亡; p38 MAPK通路阻断剂SB202190可下调Cytc、active caspase 3水平、抑制细胞凋亡。m GluR4激动剂通过p38 MAPK通路调控SH-SY5Y细胞的存活。结论:m GluR4激动剂通过p38 MAPK通路调控SH-SY5Y细胞的存活。     

天麻素对缺血缺氧大鼠星形胶质细胞MCP1表达的影响

目的:通过建立新生大鼠缺血缺氧脑损伤(HIBD)模型和体外星形胶质细胞系TNC1细胞糖氧剥夺(OGD)模型，从体内体外探索天麻素(GAS)对星形胶质细胞缺血缺氧损伤后单核细胞趋化因子1(MCP1)表达的影响。方法:将星形胶质细胞TNC1随机分为对照组(control)、糖氧剥夺组(OGD)、糖氧剥夺+天麻素(OGD+G)。将3 d新生SD大鼠随机分为假手术组(sham)、缺血缺氧脑损伤模型组(HIBD)、HIBD模型+天麻素干预组(HIBD+G)。使用免疫荧光双标染色和Western Blot检测各组细胞及大鼠左侧损伤皮质半暗区MCP1的表达变化。结果:体外免疫荧光染色和Western Blot结果显示，与control组相比，OGD组TNC1细胞MCP1表达明显增加(P<0.05),GAS可减轻OGD损伤后MCP1的表达(P<0.05);体内免疫荧光双标染色和Western Blot结果也显示：GAS干预显著降低了HIBD后星形胶质细胞MCP1的表达(P<0.05)。结论:GAS可以通过降低星形胶质细胞MCP1的表达对HIBD发挥保护作用。     

黄芪甲苷对氧糖剥夺复氧后星形胶质细胞上TNF-α与AQP-4表达的影响

目的:探索黄芪甲苷(astragaloside IV,ASIV)对氧糖剥夺复氧(oxygen glucose deprivation/reperfusion,OGD/R)后星形胶质细胞上肿瘤坏死因子-α(tumor necrosis facror-alpha,TNF-α)与水通道蛋-4(aquaporin-4,AQP-4)表达的影响。方法:选用第二代第1 d的星形胶质细胞。将星形胶质细胞放入缺氧培养箱(1%O_2、94%N2、5%CO_2),分别干预1、2、3、4 h,复氧24 h后用RT-PCR和Western Blot检测AQP-4和TNF-α的表达情况。将星形胶质细胞分为对照组、OGD/R组、OGD/R+0.001μmol/ml ASIV组、OGD/R+0.01μmol/ml ASIV组,用RT-PCR和免疫荧光检测AQP-4和TNF-α的表达情况。结果:RT-PCR和Western Blot结果显示:氧糖剥夺复氧后AQP-4和TNF-α的表达均增加,且AQP-4在OGD3 h时表达达到高峰,TNF-α在OGD1 h时表达达到高峰(P0.05);OGD3h+0.001μmol/ml ASIV组和OGD3 h+0.01μmol/ml ASIV组的AQP-4基因表达水平较OGD3 h组相比均明显降低(P0.05);与OGD3 h组相比,OGD3 h+0.001μmol/ml ASIV组的AQP-4蛋白表达水平明显降低(P0.05);OGD1 h+0.001μmol/ml ASIV组和OGD1 h+0.01μmol/ml ASIV组的TNF-α基因表达水平与OGD1 h组相比均明显降低(P0.05);与OGD1 h组相比,OGD1 h+0.001μmol/ml ASIV组的TNF-α蛋白表达水平明显降低(P0.05)。结论:氧糖剥夺复氧模型可以诱导星形胶质细胞上TNF-α和AQP-4的表达,且TNF-α的表达先于AQP-4;黄芪甲苷可能是通过下调TNF-α的表达来影响AQP-4的表达,进而影响细胞水肿。     

天麻素对少突胶质前体细胞糖氧剥夺损伤的保护作用

目的:探索天麻素对少突胶质前体细胞系OLN93细胞糖氧剥夺(OGD)损伤后血管紧张素受体1(AT1)、NADPH氧化酶2(NOX2)和组织沉默调节蛋白3(Sirt3)表达的影响。方法:将OLN93细胞随机分为对照组(control)、糖氧剥夺组(OGD)和天麻素预处理组(OGD+G),通过CCK-8观察天麻素对细胞活力的影响；免疫荧光双标染色与Western Blot检测肿瘤坏死因子-α(TNF-α)及AT1/NOX2/Sirt3的表达变化。结果:CCK8-kit结果显示：与control相比，OGD组OLN93细胞活力明显下降(P<0.05);GAS组OLN93细胞糖氧剥夺损伤后细胞活力明显有所提高(P<0.05)。Western Blot结果显示：与control相比，OGD组OLN93细胞TNF-α、AT1、NOX2蛋白表达明显增加(P<0.05);GAS明显降低OLN93细胞OGD损伤后TNF-α、AT1、NOX2蛋白表达，进一步促进Sirt3蛋白表达(P<0.05)。免疫荧光双重染色结果显示：GAS预处理显著降低OLN93细胞OGD损伤后TNF-α、A...     

天麻素对激活小胶质细胞miR-124和miR-155表达的影响

目的:研究天麻素对激活的小胶质细胞miR-124和miR-155表达含量的影响。方法:分为体内实验和体外实验,体内实验：将新生3 d的SD大鼠随机分为假手术组(sham)、缺血缺氧模型组(HIBD)、缺血缺氧+天麻素干预组(G+HIBD),体外实验：将BV2细胞随机分为对照组(control)、氧糖剥夺组(OGD)、天麻素+氧糖剥夺组(G+OGD),real time RT-PCR检测大鼠胼胝体及BV2细胞miR-124和miR-155的表达变化。结果:体内、体外real time RT-PCR结果显示：与sham、control组相比,HIBD组大鼠胼胝体及OGD处理的BV2细胞、OGD组的miR-155表达水平升高,miR-124表达水平降低(P<0.05);使用天麻素干预后,miR-155表达水平降低,miR-124表达水平升高(P<0.05)。结论:天麻素能促进激活的小胶质细胞miR-124的表达,抑制miR-155表达,从而发挥其神经保护作用。     

自噬参与低氧预处理抗PC12细胞糖氧剥夺损伤过程

目的:观察低氧预处理(HPC)对氧糖剥夺(OGD)损伤的PC12细胞的作用,同时探讨自噬在其中的作用。方法:培养的PC12细胞按照处理因素分为6组:对照组、HPC模型组、3-甲基腺嘌呤(3-MA)组、低氧预处理后氧糖剥夺(HPC+OGD)组、3-甲基腺嘌呤预处理后行低氧预处理及氧糖剥夺(3-MA+HPC+OGD)组和OGD组。CCK-8检测细胞存活率;caspase-3活性检测试剂盒检测酶活性;TUNEL染色和流式细胞术检测细胞凋亡的情况;Western blot法检测自噬相关蛋白LC3、beclin-1和凋亡相关蛋白caspase-3的表达。结果:与对照组相比,OGD组细胞存活率明显降低;与3-MA+HPC+OGD组及OGD组相比,HPC+OGD组细胞存活率明显升高(P0.05)。与对照组相比,OGD组细胞的caspase-3酶活性明显升高;与3-MA+HPC+OGD组及OGD组相比,HPC+OGD组的caspase-3酶活性明显降低(P0.05)。与对照组相比OGD组细胞凋亡明显增多;HPC+OGD组与OGD组相比凋亡明显减少(P0.05)。此外,与对照组相比,OGD组的激活型caspase-3蛋白水平明显升高(P0.05);且HPC+OGD组与OGD组相比激活型caspase-3蛋白水平明显减少而LC3、beclin-1的蛋白水平明显升高(P0.05)。结论:HPC抗OGD损伤的机制可能与其激活细胞自噬有关。     

褪黑素促进糖氧剥夺诱导的BV2小胶质细胞向M2极化

目的:研究褪黑素处理对糖氧剥夺(OGD)诱导的小鼠小胶质细胞系BV2细胞功能的影响。方法:BV2细胞分为对照组(control)、氧糖剥夺组(OGD)和褪黑素处理组(OGD+Mel)。利用流式细胞仪检测各组细胞CD16/32和兔多克隆抗体CD206的表达,real time RT-PCR方法检测BV2细胞中iNOS和Arg-1 mRNA表达,利用酶联免疫吸附试验(ELISA)方法检测BV2细胞培养上清中IL-1β和IL-4的水平。结果:OGD诱导可促进BV2细胞向M1极化,表现为CD16/32上调、iNOS mRNA表达增加,培养上清中IL-1β水平升高;经过褪黑素处理后,CD16/32、iNOS mRNA,培养上清中IL-1β表达下降,CD206、 Arg-1和IL-4表达增加。结论:OGD可诱导BV2细胞向M1方向分化,而褪黑素可使OGD诱导的BV2细胞向M2极化。     

阿利吉仑对氧糖剥夺损伤的SH-SY5Y细胞的保护作用及可能机制

目的:探讨阿利吉仑对氧糖剥夺(OGD)损伤的神经母细胞瘤SH-SY5Y细胞的保护作用及可能机制。方法:将SH-SY5Y细胞随机分为对照组、OGD组及阿利吉仑低、中和高剂量(5.0、10.0和20.0μmol/L)组。CCK-8法检测细胞活力;ELISA检测兴奋性氨基酸转运蛋白2(EAAT2/GLT-1)、兴奋性氨基酸转运蛋白3(EAAT3/EAAC1)、兴奋性氨基酸转运蛋白4(EAAT4)、内皮素1(ET-1)和S100钙结合蛋白β亚基(S-100β)的表达;Hoechst 33258染色观察SH-SY5Y细胞形态变化;乳酸(LD)测试盒和超微量Na~+-K~+-ATP酶测试盒检测LD含量和Na~+-K~+-ATPase活性。结果:OGD损伤4 h时,细胞相对活力不足60%,因此4 h可作为后续实验OGD造模时间。与对照组相比,OGD组的GLT-1、EAAC1和EAAT4表达显著下调(P0.05),ET-1和S-100β的表达显著上调(P0.05);与OGD组相比,阿利吉仑组的GLT-1、EAAC1和EAAT4表达呈剂量依赖性上调, ET-1和S-100β表达呈剂量依赖性下调(P0.05)。Hochest 33258染色结果表明,阿利吉仑可明显减少OGD引起的SH-SY5Y细胞凋亡。与对照组相比,OGD组的LD含量显著升高(P0.05),Na~+-K~+-ATPase活性显著降低(P0.05);与OGD组相比,阿利吉仑组的LD含量呈剂量依赖性降低,Na~+-K~+-ATPase活性呈剂量依赖性升高(P0.05)。结论:阿利吉仑对OGD损伤的SH-SY5Y细胞有一定的保护作用,其机制可能与阿利吉仑上调GLT-1、EAAC1和EAAT4的水平,提高Na~+-K~+-ATPase的活性,下调ET-1和S-100β的表达及LD含量有关。     

孕酮对氧糖剥夺损伤的PC12细胞的保护作用

目的: 观察孕酮(PROG)对氧糖剥夺(OGD)PC12细胞活力及葡萄糖转运蛋白3(GLUT3)表达的影响,在培养细胞证明孕酮抗缺氧缺血损伤的神经保护作用。方法: 神经生长因子诱导分化良好的PC12细胞,随机分为3组:(1)正常对照组:常规培养,不进行OGD处理;(2)OGD组:无糖培养基、低氧环境(95%N₂和5%CO₂持续通气)进行OGD 30 min处理,然后复氧复糖培养24 h;(3)PROG +OGD组:培养液含终浓度为10 nmol/L 孕酮,余同OGD组。WST-8法检测各组PC12细胞的活力;检测培养液中乳酸脱氢酶(LDH)的活性,判断细胞损伤的程度;反转录-聚合酶链反应(RT-PCR)检测PC12细胞GLUT1 mRNA和GLUT3 mRNA表达;Western blotting法检测PC12细胞GLUT3蛋白表达。结果: 孕酮可以减轻OGD引起的PC12细胞肿胀,降低LDH漏出,减轻细胞损伤,显著增加OGD组PC12细胞的活力(P<0.01)。RT-PCR显示PROG +OGD组GLUT3 mRNA的表达明显高于OGD组(P<0.05),Western blotting显示PROG +OGD组GLUT3蛋白的表达显著高于OGD组(P<0.01)。结论: 孕酮对体外培养OGD损伤的PC12细胞具有保护作用,其机制可能与上调GLUT3蛋白的表达有关。     

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.