热休克蛋白70对未成熟心肌细胞功能的影响

目的探讨热休克蛋白70(HSP70)对未成熟心肌细胞功能的影响。方法健康新生长耳大白兔随机分为2组。对照组(C):腹腔注射生理盐水0.4ml,注射后24h取离体心脏,常规建立Langendorff离体心脏灌注模型,灌注15min转为工作心15min后停灌45min,恢复灌注15min改为工作心30min。实验组(E):腹腔注射重酒石酸去甲肾上腺素,24h后取离体心脏,方法同对照组。测定心肌细胞中HSP70含量及相关生化指标并作统计学处理比较。结果HSP70含量E组明显高于C组;E组ATP含量、SOD活性方面均优于C组(P<0.01),MDA含量、CK、LDH漏出率方面均低于C组(P<0.01),心肌线粒体Ca2+-ATPase活性、[ATP]m均优于C组(P<0.01),心肌细胞内Ca2+含量、心肌线粒体Ca2+含量低于C组(P<0.01)。结论HSP70对缺血再灌注未成熟心肌细胞功能具有明显的保护作用。     

金属硫蛋白对离体心脏NO、NOS表达的影响

目的探讨金属硫蛋白(MT)对离体心脏一氧化氮(NO)、一氧化氮合酶(NOS)的影响。方法Wistar大鼠16只 ,分为2组 :对照组(C,n=8) ,腹腔注射蒸馏水0.5ml24h后取离体心脏灌注HTK心脏保护液 ,4℃保存3h后建立Langendorff灌注模型 ,灌注KH液2h ;实验组 (E ,n=8)腹腔注射3.6%Zn SO4(1.5ml/kg)24h后取离体心脏 ,处理方法同C组。以心肌MT含量、NO、NOS的含量、心肌生化指标作为观察指标。结果MT含量E组明显较C组增高 (P<0.01) ,NO、NOS的含量E组较C组明显增多(P<0.01) ,生化指标E组明显优于C组。结论心肌MT表达影响NO、NOS的表达 ,可能是MT发挥心肌保护作用的因素之一。     

去甲肾上腺素预处理对大鼠缺血再灌注心肌的延迟性保护作用

目的观察去甲肾上腺素预处理后热休克蛋白(HSP70)在心肌细胞的表达以研究去甲肾上腺素预处理对大鼠离体缺血再灌注损伤心肌的延迟性保护作用.方法24只Sprague-Dawley大鼠随机分为对照组(n=8)及实验组(E112h及E24h组),实验组按腹腔注射去甲肾上腺素到建立Langendorff灌注模型的时间分为E12h及E24h组,每组各8只.分别检测LDH及CK的漏出量、心肌ATP含量及心功能指标(LVSP及±dp/dt max),用Westem Blot法测量心肌组织中HSP70的含量.结果E12h组HSP70表达较对照组增高,E24hHSP70表达最高.E24h组LDH与CK含量较其它两组低,而ATP较其它两组高(P＜0.01),心功能改善.E12h与对照组相应各指标(LDH、CK、ATP、LVSP及±dp/dt max)间差异无显著性.结论去甲肾上腺素预处理24h后HSP70高表达对大鼠离体缺血再灌注心肌有延迟性保护作用.     

热休克蛋白70对离体心脏心肌间质的保护作用

目的探讨热休克蛋白70（HSP70）对大鼠离体心脏心肌间质的影响。方法Wistar大鼠16只，分为2组：对照组（C，n=8），腹腔注射生理盐水0．4ml，24h后取离体心脏灌注HTK心脏保护液，4℃保存3h后建立Langendorff灌注模型，灌注KH液2h；实验组（E，n=8），腹腔注射重酒石酸去甲肾上腺素，24h后取离体心脏，方法同对照组。以心肌细胞中HSP70含量、血流动力学指标、心肌组织羟脯氨酸（HP）、内皮索（ET）含量和心肌超微结构等作为观察指标。结果HSP70含量E组与C组比较明显增高；E组心功能恢复方面优于C组（P〈0．05），HP含量优于C组（P〈0．01），ET含量低于C组（P〈0．01），心肌超微结构损伤较C组明显减轻。结论HSP 70对供心心肌间质具有保护作用。     

Zn~(2+)预处理对离体大鼠缺血再灌注损伤心肌的保护作用

目的 :观察Zn2 + 预处理诱导金属硫蛋白 (MT)在心肌细胞的表达及对离体大鼠缺血再灌注损伤心肌的保护作用。方法 :3 2只Sprague Dawley大鼠随机分为对照组及实验组 ,实验组按腹腔注射ZnSO4 到建立再灌注模型的时间分为E1 2h,E2 4h及E4 8h组 ,每组各 8只。检测LDH ,CK、ATP含量及心功能指标 (LVSP与±dp/dtmax) ,用1 0 9Cd/血红素饱和法测量心肌组织中金属硫蛋白的含量。结果 :E2 4h及E4 8h组MT表达量较对照组高 (P <0 .0 1) ,E1 2h组虽有表达 ,但其与对照组间差异无显著性。E2 4h及E4 8h组LDH与CK较其它二组低 ,而ATP较其它二组高 ,心功能指标较其它二组改善。E1 2h组及对照组之间相应各指标 (LDH ,CK、ATP、LVSP与±dp/dtmax)间差异无显著性 (P >0 .0 5 )。结论 :Zn2 + 预处理可诱导MT在心肌细胞的表达而对离体大鼠缺血再灌注损伤心肌具有保护作用。     

牛磺酸对烧伤大鼠心肌细胞浆、线粒体及核内钙含量变化的干预作用

目的探讨大鼠烧伤后心肌细胞胞浆、线粒体、核内钙含量变化及牛磺酸的干预作用。方法大鼠30%Ⅲ度烫伤(烧伤组),伤后即刻腹腔注射2%牛磺酸液(200mg/kg体质量,牛磺酸治疗组)。两组均于伤后1、3、6、12、24、48h检测核膜Ca2 -ATP酶活性,核、胞浆、线粒体钙含量。37℃假烫伤为对照组,1h后检测上述各项为对照值。结果大鼠伤后心肌细胞核Ca2 -ATP酶活性明显下降,而核内、胞浆、线粒体钙含量均明显升高。牛磺酸治疗组核Ca2 -ATP酶活性明显高于烧伤组,胞浆、线粒体、核钙含量均明显低于烧伤组。结论大鼠烧伤后心肌细胞胞浆、线粒体、核钙含量均明显升高,牛磺酸具有较好的拮抗作用。     

脂多糖诱导脓毒症小鼠心肌细胞及线粒体自噬

目的探讨脓毒症心肌损伤时心肌细胞及线粒体自噬水平的变化。方法雄性C57BL/J小鼠随机分为空白对照组(NC)、LPS处理6、12、24、36 h组。LPS处理组小鼠腹腔注射10 mg/kg LPS建立脓毒症模型,空白对照组注射等量生理盐水。分别于以上时间点处死小鼠,收集血液及心脏组织,提取心肌组织胞质蛋白、线粒体及线粒体蛋白,另取心肌组织进行冰冻切片;应用ELISA试剂盒测定血清心肌肌钙蛋白I(c Tn I)含量变化;JC-1染色结合荧光酶标仪检测LPS刺激后不同时间点线粒体膜电位变化;Western blot法检测自噬相关蛋白微管相关蛋白1轻链3(LC3)、PTEN诱导激酶1(pink1)、E3泛素连接酶帕金森病蛋白(parkin)水平;免疫荧光组织化学染色检测心肌组织LC3、pink1/parkin蛋白的定位及其在不同处理组的表达差异。结果与对照组相比,LPS诱导的脓毒症小鼠血清c Tn I在6 h即开始明显升高;LPS处理组线粒体膜电位下降,在LPS 12 h组最低;细胞自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ在腹腔注射LPS 12 h明显升高,然后逐渐下降;pink1/parkin在腹腔注射LPS 6 h明显升高,然后逐渐下降。结论脓毒症心肌损伤时心肌细胞及线粒体自噬水平升高,自噬应激可能更早发于心肌线粒体。     

Urocortin-I预处理对离体大鼠心肌线粒体呼吸功能及呼吸酶活性的影响

目的:探讨urocortin-I预处理对离体大鼠缺血再灌注心肌线粒体呼吸功能及酶活性的影响,观察心肌细胞ATP含量的变化。方法:(1)健康雄性SD大鼠随机分为4组:正常组(Nor组)、缺血再灌注组(IR组)、urocortin-I预处理组(Ucn I组)、5-羟葵酸(5-HD)拮抗urocortin-I组(5-HD+Ucn I组)。采用Langendorff装置建立大鼠离体心脏缺血再灌注模型。分别于平衡末(T_1)、缺血前(T_2)及再灌注末(T_3)分离、提取心肌线粒体,测定各组线粒体呼吸功能及呼吸酶活性。(2)利用MPA离体心脏灌注装置分离成年大鼠心肌细胞,将分离培养24 h后同批次的心肌细胞随机分为正常组(Nor组)、缺氧复氧组(I/R组)、urocortin-I预处理组(Ucn I组)、5-HD拮抗urocortin-I组(5-HD+Ucn I组)。建立缺氧复氧模型,于复氧末用高效液相色谱法检测各组心肌细胞ATP含量。结果:T_3时点除Nor组外,其余各组与T_1、T_2时点相比呼吸功能(3态呼吸速率、呼吸控制率)及琥珀酸氧化酶、NADH氧化酶、细胞色素C氧化酶活性均明显下降(P0.05);T_3时Ucn I组心肌线粒体的呼吸功能及呼吸酶活性明显优于5-HD+Ucn I组及IR组(P0.05),但次于Nor组(P0.05);T_3时5-HD+Ucn I组心肌线粒体的呼吸功能及呼吸酶活性(琥珀酸氧化酶、NADH氧化酶)较IR组好(P0.05),但2组间细胞色素C氧化酶活性差异无统计学显著性;T_1、T_2时点各组组内及组间呼吸功能及3种呼吸酶活性的差异无统计学显著性。心肌细胞实验结果显示,复氧末Nor组ATP含量较其余各组均高(P0.01);I/R组和5-HD+Ucn I组心肌细胞的ATP含量较Ucn I组低(P0.05);此外,5-HD+Ucn I组心肌细胞的ATP含量较I/R组高(P0.05)。结论:Ucn I预处理可减轻缺血再灌注对心肌线粒体呼吸功能及呼吸酶活性的干预,保证了缺氧/复氧后心肌ATP的含量。     

镁对缺血再灌注心肌线粒体合成ATP能力的影响

我们曾经观察到心肌缺血再灌注过程中心肌线粒体Ca~(2+)-ATP酶活性降低,Ca~(2+)大量沉积于线粒体内.含Mg~(2+)溶液灌流对此具有逆转作用。心肌线粒体是心肌细胞进行氧化磷酸化、合成ATP的场所,大量的钙盐沉积,有可能抑制ATP的合成,因此,我们进一步观察含Mg~(2+)溶液灌注对缺血再灌注心肌线粒体合成ATP的影响。实验采用Wistar大鼠离体等容收缩心脏,行Langendorff灌流,常规K-H液预灌流30分钟行低灌流缺血(0.2ml/min,60分钟后,常规K-H液再灌)注30分钟.根据缺血期准注K-H液中Mg~(2+)浓度不同,实验分为0、1.2及15μmol三组(n均为5)。灌流结束后差速离心分离心肌线粒体,加入含有底物ADP     

休克再灌注后早期UCP—2对线粒体膜电位的影响作用

目的通过观察休克-再灌注大鼠肠上皮、骨骼肌、心肌细胞线粒体UCP-2的变化规律及其对线粒体膜电位(MP)的影响,探讨休克再灌注后不同组织线粒体功能发生差异性损伤的机制.方法取150-220gWistar大鼠,复制休克再灌注模型,差速离心法游离肠上皮、心肌、骨骼肌细胞线粒体,用Westernblot法测量UCP-2含量,用荧光分光光度计法测量线粒体膜电位.实验分组如下对照组(假手术)、单纯休克组(40mmHg90min)、再灌注(单纯休克+回输全部失血量及2倍失血量的晶体液)2h组、再灌注6h组、再灌注12h组、再灌注24h组.结果(1)失血性休克90min后,3种组织线粒体UCP-2均有升高,与对照组相比有显著差异(P＜0.05),肠上皮线粒体UCP-2在再灌注后24h内均高于对照组(P＜0.05),心肌线粒体UCP-2于再灌注后2h高于对照组,但再灌注后6h与对照组相比无显著差异(P＜0.05),骨骼肌线粒体UCP-2于再灌注后6h高于对照组,但再灌注后12h与对照组相比无显著差异(P＜0.05);(2)在测量MP的各组实验中,加入UCP-2抑制剂GDP后MP均高于无GDP处理组(P＜0.01);(3)休克及再灌注后24h内,肠上皮细胞线粒体MP均比对照组降低,心肌线粒体MP于休克及再灌注后2h低于对照组,于再灌注后6h与对照组相比无显著差异,骨骼肌线粒体MP于休克及再灌注后2h低于对照组,于再灌注后6h与对照组相比无显著差异,但再灌注后12-24h骨骼肌线粒体MP又低于对照组(P＜0.05).讨论近来认为,UCP-2使质子泄漏引起MP降低,其结果导致氧化磷酸化解偶联,ATP合成效率降低,同时能够降低4态呼吸时线粒体内ROS的产生.本实验也证实,加入GDP抑制UCP-2的质子转运活性,线粒体MP增高.休克导致线粒体UCP-2表达上调,再灌注后UCP-2水平逐渐恢复,但不同组织中UCP-2的变化规律不同,心肌细胞线粒体UCP-2水平恢复较快,骨骼肌及肠上皮恢复较慢,这可能与休克时不同组织的缺血程度不同有关,其结果与MP及线粒体功能的恢复可能有关,这可能是导致休克再灌注后不同组织线粒体功能发生差异性损伤的机制之一.结论本实验结果提示UCP-2可能介导休克-再灌注后MP的调节,休克再灌注后,不同组织中线粒体UCP-2的变化规律不同,可能与休克再灌注后不同组织线粒体功能发生差异性损伤有关.     

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.