甘氨酸对脂多糖诱导LBP/CD14 mRNA表达的影响

目的内毒素是位于革兰氏阴性杆菌(G-)细胞壁外膜中具有高度生物学活性的以脂多糖(lipopolysaccharide,LPS)为主的成分.LBP(lipopolysaccharide-bindingprotein)/CD14(clusterofdifferentiation14)系统在识别和调控LPS作用方面起着关键作用.甘氨酸是体内固有的最简单的氨基酸,本教研室在先前的研究中发现甘氨酸对LPS的致热性有良好的拮抗作用,体外研究发现甘氨酸能破坏内毒素的构型,抑制内毒素诱导单核/巨噬细胞分泌TNF、IL-1等细胞因子.本研究是在前面研究的基础上进一步探讨甘氨酸对内毒素体内作用过程的影响,从而为研制有效的内毒素拮抗剂提供理论依据.方法用RT-PCR技术检测大鼠肝组织LBPmRNA表达水平,并在此基础上观察甘氨酸对内毒素诱导大鼠肝组织LBPmRNA表达的影响.用原位杂交方法检测小鼠腹腔巨噬细胞CD14mRNA表达水平,并在此基础上观察甘氨酸对内毒素诱导小鼠腹腔巨噬细胞CD14mRNA表达的影响.结果比较5组大鼠肝组织LBPmRNA的表达水平,内毒素组显著高于其他各组(P＜0.01),甘氨酸拮抗组显著低于内毒素组(P＜0.01),但甘氨酸拮抗组仍高于对照组.比较5组小鼠腹腔巨噬细胞的CD14mRNA表达水平,内毒素组小鼠腹腔巨噬细胞CD14mRNA表达水平显著高于其他各组(P＜0.01),甘氨酸拮抗组显著低于内毒素组(P＜0.01).结论(1)内毒素可诱导大鼠肝组织LBPmRNA的表达.(2)甘氨酸可抑制内毒素诱导大鼠肝组织LBPmRNA的表达.(3)内毒素可诱导小鼠腹腔巨噬细胞CD14mRNA的表达.(4)甘氨酸可抑制内毒素诱导小鼠腹腔巨噬细胞CD14mRNA表达.     

HIF-1α和MMP-2在大鼠脑出血灶周脑组织中的表达

目的：通过动态观察脑出血灶周脑组织中缺氧诱导因子(HIF)-1α和基质金属蛋白酶(MMP)-2的表达,探讨HIF-1α和MMP-2表达之间的联系。方法：50只SD大鼠随机分成假手术组和脑出血模型组,分别于术后6 h,24 h,72 h,7 d,21 d处死大鼠。RT-PCR方法检测脑出血灶周组织HIF-1α和MMP-2 mRNA的表达及免疫组织化学方法检测HIF-1α和MMP-2蛋白的表达,并对HIF-1α和MMP-2的表达进行相关分析。结果：模型组术后灶周组织出现HIF-1α, MMP-2 mRNA表达上调和大量黄色的HIF-1α,MMP-2蛋白阳性细胞,于24~72 h达高峰。HIF-1α mRNA及其蛋白表达分别与MMP-2 mRNA及其蛋白表达呈正相关(分别r=0.588, P=0.002; r=0.765, P＜0.001)。结论：脑出血灶周脑组织中HIF-1α和MMP-2的表达上调,且HIF-1α可能调控MMP-2的表达。     

甘氨酸对脂多糖诱导LBPmRNA表达的影响

目的:了解甘氨酸(GLY)对脂多糖(LPS)诱导脂多糖结合蛋白(LBP)mRNA表达的影响。方法:用RT-PCR方法检测LPS作用下大鼠肝组织LBPmRNA的表达水平,并在此基础上观察甘氨酸对LPS诱导大鼠肝组织LBPmRNA表达的影响。结果:LPS组大鼠肝组织LBPmRNA表达高于control组(P＜0.01),GLY+LPS组大鼠肝组织LBPmRNA表达低于LPS组(P＜0.01)。结论:LPS可诱导大鼠肝组织LBPmRNA的表达,甘氨酸能抑制LPS诱导大鼠肝组织LBPmRNA的表达。     

右美托咪啶减轻急性肺损伤的机制研究

目的探讨右美托咪啶(Dex)对小鼠急性肺损伤(ALI)的影响及潜在作用机制。方法将32只C57BL/6雄性小鼠随机分为空白对照组(Sham组)、右美托咪啶组(Dex组)、脂多糖组(LPS组)和药物干预组(LPS+Dex组)。LPS组和LPS+Dex组小鼠通过腹腔注射LPS(10 mg/kg)构建小鼠ALI模型,Dex组及LPS+Dex组小鼠腹腔注射Dex(40μg/kg),Sham组和LPS组注射等剂量生理盐水。LPS注射12 h后,采用HE染色比较各组小鼠肺损伤状况;qRT-PCR检测各组小鼠肺组织中促炎性细胞因子IL-1β、TNF-α、IL-6和MCP-1 mRNA的表达;Western blot检测各组小鼠肺组织中GPX4、COX2和转录因子红系2相关因子2(Nrf2)蛋白的表达。结果与Sham组相比,LPS组小鼠肺损伤评分明显升高(P0.05),促炎性细胞因子IL-1β、TNF-α、IL-6和MCP-1mRNA表达水平均明显升高(P0.05),肺组织铁死亡标志物GPX4蛋白表达水平明显降低(P0.05),COX2蛋白表达水平则明显升高(P0.05),肺组织中Nrf2蛋白表达水平明显降低(P0.05)。与LPS组相比,LPS+Dex组小鼠肺损伤评分明显降低(P0.05),IL-1β、TNF-α、IL-6和MCP-1 mRNA表达水平明显降低(P0.05),GPX4的蛋白表达水平明显升高(P0.05),COX2蛋白表达水平则明显降低(P0.05),Nrf2的蛋白表达水平也明显升高(P0.05)。结论 Dex可能通过激活Nrf2抑制小鼠肺组织铁死亡,进而发挥肺保护作用。     

深低温停循环下幼猪脑皮质HIF-1与NGB的表达及作用

目的:观察深低温停循环(DHCA)下幼猪脑皮质缺氧诱导因子1(HIF-1)与神经珠蛋白(NGB)的表达变化。方法:五指山猪共15头随机分成体外循环组(CPB组)、深低温停循环组(DHCA组)与深低温停循环选择性顺行脑灌注组(SACP组)。建立体外循环后,DHCA组降温至18℃后停循环40 min,SACP组停循环后经无名动脉顺行脑灌注40 min。复灌180 min后取脑皮质组织行HE染色镜检,Western blot及real-time PCR检测HIF-1α、NGB蛋白表达及mRNA水平。结果:HE染色显示与DHCA组相比SACP组脑组织损伤显著减轻。复灌180min后DHCA组及SACP组脑皮质中HIF-1α的蛋白及mRNA水平均显著高于CPB组(P0.05),同时SACP组动物脑组织HIF-1α的蛋白及mRNA水平显著高于DHCA组(P0.05)。复灌180 min后DHCA组及SACP组脑皮质中NGB的蛋白及mRNA水平均显著高于CPB组(P0.05),同时SACP组动物脑组织NGB的蛋白及mRNA水平显著高于DHCA组(P0.05)。结论:HIF-1与NGB的表达上调参与了脑组织对DHCA脑损伤的反应机制,并可能是SACP脑保护作用的机制之一。     

灯盏花素对大鼠脑缺血后细胞间粘附分子-1及其mRNA表达的影响

目的:灯盏花素对大鼠脑缺血后细胞间粘附分子-1(ICAM-1)及其mRNA表达的影响。方法:复制大鼠大脑中动脉闭塞/再灌注模型,应用RT-PCR及免疫组织化学的方法,观察各组大鼠脑缺血后细胞间粘附分子-1 mRNA及其蛋白的表达。结果:ICAM-1在假手术组大鼠脑组织呈低表达;单纯缺血组(缺血90 min)ICAM-1表达上调(P＜0.05);缺血再灌注组(缺血90 min再灌24 h)脑组织ICAM-1表达显著高于假手术组和单纯缺血组(P＜0.01);灯盏花素治疗组于相同时限ICAM-1表达与单纯缺血、缺血再灌注组相比显著下调(P＜0.01)。大鼠脑组织ICAM-1 mRNA在假手术组呈低表达;单纯缺血组ICAM-1 mRNA水平显著上调(P＜0.01);药物治疗组于相同时限ICAM-1 mRNA水平显著低于单纯缺血组及缺血再灌注组(P＜0.01)。结论:灯盏花素下调细胞间粘附分子-1mRNA及其蛋白的表达,减轻缺血后再灌注损伤,从而发挥脑保护作用。     

Wortmannin对急性肺损伤小鼠肺组织IL-1β和TNF-α表达的影响

目的研究Wortmannin对急性肺损伤模型小鼠肺组织白介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)表达的影响。方法 30只昆明小鼠随机分为正常对照组、急性肺损伤组和Wortmannin处理组。采用腹腔注射LPS(10 mg/kg)建立小鼠急性肺损伤模型,对照组腹腔注射同体积的生理盐水,Wortmannin处理组则于造模前2 h腹腔注射Wortmannin(1.4 mg/kg)。LPS注射后6 h处死大鼠,计算肺组织湿/干重(W/D)比值,Western blot方法检测三组小鼠肺组织内IL-1β和TNF-α蛋白的表达变化,RT-PCR方法检测三组小鼠肺组织内IL-1βmRNA和TNF-αmRNA的表达变化。结果急性肺损伤组小鼠肺组织IL-1β和TNF-α蛋白及mRNA表达水平显著上升,显著高于正常对照组(0.05);相比于急性肺损伤组小鼠,Wortmannin处理组小鼠肺组织IL-1β和TNF-α蛋白及mRNA表达水平显著降低(0.05)。结论 Wortmannin能抑制急性肺损伤小鼠肺组织IL-1β和TNF-α表达。     

癫痫大鼠海马NMDA受体亚基NR2A和BDNF mRNA水平的检测

目的 通过锂-匹罗卡品癫痫模型(ithium-pilocarpine seizures rats model of epilepsy,LPS),研究NMDA受体亚基NR2A、BDNF mRNA的表达,探讨NR2A、BDNF在LPS中的作用.方法 建立氯化锂-匹罗卡品大鼠模型,运用原位杂交技术检测致痫后各组不同时间点海马CA1、CA3及DG区NR2A与BDNF mRNA的表达.结果 LPS海马NR2A、BDNF mRNA在各观察时间点及部位模型组与正常对照组比较均有明显上调,且有显著统计学差异(P＜0.05).模型组NR2A mRNA的表达上调7 d达峰值(P＜0.05);而BDNF mRNA表达上调14 d达峰值.VPA干预组NR2A mRNA在大鼠海马不同时间及部位(除1 d的CA3区)的表达较模型组明显下调(P＜0.05);BDNF mRNA在大鼠海马不同时间及部位(除28 d的DG区)的表达较模型组明显下调(P＜0.05).结论 锂-匹罗卡品腹腔注射可诱导大鼠海马NR2A和BDNF mRNA的表达明显上调;NR2A mRNA表达的增强可能是诱导调控BDNF mRNA表达增强的重要机制之一,说明NMDA受体亚基NR2A可能成为抑制癫痫发作的新靶点.     

妊娠期脂多糖接触激活体内TLR4信号通路引起胚胎脑内炎症应激

目的: 观察孕鼠腹腔脂多糖 (LPS) 注射后母体外周血单个核细胞 (PBMNC) 上Toll样受体4 (TLR4) 信号通路激活情况和胚胎脑组织中炎症性细胞因子的水平。方法: 10.5 d孕鼠腹腔注射LPS 0、3、6、12、24、48 和72 h后,通过蛋白免印迹测定孕鼠PBMNC 上TLR4、分化抗原14 (CD14)、髓样分化蛋白2 (MD-2)、p65和p50蛋白水平,Luminex 100免疫荧光法测定孕鼠血清、羊水、脑组织及胚胎脑组织细胞因子TNF-α、IL-1β和IL-10蛋白水平,实时RT-PCR测定孕鼠PBMNC和胚胎脑组织TNF-α、IL-1β和IL-10 mRNA水平。结果: LPS腹腔注射诱导激活孕鼠PBMNC上TLR4信号通路,TLR4、CD14和MD-2蛋白水平短暂增高,核因子κB (NF-κB) 激活片段p65 和p50蛋白水平增高,TNF-α、IL-1β和IL-10 mRNA水平增高(P<0.01);孕鼠外周血和羊水TNF-α、IL-1β和IL-10水平短暂增高(P<0.01),脑内IL-1β蛋白水平短暂升高(P<0.01);胚胎脑内TNF-α和IL-1β蛋白和mRNA水平显著增高(P<0.01)。结论: 妊娠期母体LPS接触可激活母体外周免疫细胞TLR4信号通路,引发胚胎脑内炎症应激状态,可能增加出生后相关疾病的发生风险。     

白细胞介素-18在内毒素诱导的老年大鼠记忆障碍中的作用

目的:探讨白细胞介素-18(IL-18)在内毒素(LPS)诱导的老年大鼠记忆障碍中的作用。方法:16只老年雄性Sprague-Dawley(SD)大鼠随机分为内毒素组和生理盐水对照组,在被动回避训练结束后,立即经腹腔注射LPS250μg/kg或等体积的生理盐水,24h后记录大鼠从明室进入暗室的潜伏期作为记忆成绩。记忆功能测试结束后立即取海马组织,分别用逆转录-多聚酶链式反应和酶联免疫吸附实验测定IL-18mRNA和IL-18蛋白的表达,并检测其中半胱氨酸天冬氨酸蛋白酶-1(caspase-1)的活性。另分批实验,将16只老年SD大鼠随机分为IL-18组和生理盐水对照组,同样在训练后经脑室注射IL-18(100ng)或等体积的生理盐水,24h后记录大鼠进入暗室的潜伏期。结果:LPS组大鼠进入暗室的潜伏期较对照组明显缩短,差异显著(P0.01);并且LPS组海马组织IL-18mRNA和IL-18蛋白表达水平和caspase-1酶活性均明显高于对照组,差异显著(P0.01);IL-18组大鼠进入暗室的潜伏期明显短于对照组,差异显著(P0.05)。结论:IL-18可能参与了LPS诱导的老年大鼠记忆障碍。     

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.     

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.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.