联合成像系统研究心肌细胞微区力学及细胞内钙离子变化

目的 使用联合成像系统在亚细胞水平研究心肌细胞微区力学性质及细胞内钙离子的快速变化,并探讨两者之间的关系.方法 应用原子力显微镜(AFM)和激光扫描共聚焦显微镜(LSCM)组合成的联合成像系统对急性分离的大鼠单个心室肌细胞微区力进行检测,同时测定细胞内钙离子浓度的变化,分析细胞微区力和细胞内钙离子浓度变化两者之间的相关性.结果 观察到心肌细胞内钙离子浓度的快速变化引起了钙波的传递.通过AFM微悬臂的偏转可以计算出细胞微区力,细胞微区力的大小与细胞内钙离子浓度呈正相关(r=0.701,P=0.003).结论 联合成像系统可以同步研究心肌细胞微区力学及细胞内分子快速变化,进而在亚细胞水平研究心脏疾病的发病机制.     

急性心肌梗死患者抗心肌肌球蛋白重链抗体诱导心肌细胞凋亡

目的：探讨抗心肌肌球蛋白重链抗体（AMHCA）诱导心肌细胞凋亡的作用及其机制。方法：从急性心肌梗死患者血清中提取AMHCA,研究其对成年大鼠心肌细胞凋亡的影响。DNA末端标记膜联蛋白-V/PI复染色法观察和测量心肌细胞凋亡。分别用免疫印迹、膜片钳和激光共聚焦法检测凋亡相关蛋白P53和Bcl-2以及第二信使钙离子的表达和浓度。结果：AMHCA诱导的心肌细胞凋亡具有剂量依赖性。在此过程中,促凋亡的核蛋白P53促进心肌细胞凋亡,而抑凋亡的胞质蛋白Bcl-2 抑制心肌细胞凋亡。同时,胞内钙离子浓度升高,而L型钙通道则没有影响。结论：急性心肌梗死患者体内的AMHCA可能是一种新的起动因子,可诱导心肌细胞凋亡。     

超声微泡介导转染FKBP12．6基因对小鼠n9c2（2—1）心肌细胞中Ca^2＋浓度的影响

目的：探讨超声微泡介导转染FKBP12．6基因后，对小鼠H9c2（2-1）心肌细胞中Ca^2＋浓度的影响。方法：将pcDNA3．1-FKBP12．6质粒与白蛋白包裹微泡造影剂混合，经超声转染H9c2（2-1）细胞后，通过倒置显微镜观察心肌细胞生长状况的变化；激光共聚焦显微镜检测细胞内Ca^2＋浓度的变化；免疫组织化学方法检测FKBP12．6蛋白的表达。结果：超声触发微泡破裂转染的FKBP12．6基因可在心肌细胞高效表达，细胞生长良好。高表达FKBP12．6的心肌细胞中，总的钙离子浓度增加。结论：超声微泡介导FKBP12．6基因转染心肌细胞，可以明显增加心肌细胞中的Ca^2＋浓度，心肌细胞的收缩能力增强。     

缬沙坦对心肌梗死后心衰大鼠线粒体凋亡的影响及可能机制

目的 探讨缬沙坦对心肌梗死后心衰大鼠线粒体凋亡的影响及可能机制。方法 采用左前降支冠状动脉结扎法制备心衰大鼠模型，将大鼠随机分为假手术组(Sham)、模型组(HF)、缬沙坦治疗组(ARNI);超声检测各组大鼠的心肌功能；HE染色观察心肌组织损伤；Masson染色观察心肌组织纤维化情况；激光共聚焦观察心肌钙离子荧光强度变化；TUNEL染色检测各组大鼠心肌细胞凋亡情况；定磷法检测心肌组织SERCA2a活力；Western blot检测各组大鼠心肌组织Cyto-C、Cleaved-caspase-3、Cleaved-caspase-9、SERCA2a、p-p38及p38蛋白表达。结果 与HF组比较，ARNI明显改善心衰大鼠心肌功能；减轻心肌组织病理损伤与纤维化情况；降低心肌钙离子荧光强度，抑制心肌细胞凋亡及凋亡相关蛋白Cyto-C、Cleaved-caspase-3、Cleaved-caspase-9表达，显著增强SERCA2 a活力，显著抑制p-p38蛋白表达。结论 缬沙坦可以改善心肌梗死后心衰大鼠线粒体凋亡，可能与通过介导p38MAPK-SERCA2信号通路活性调节钙离子超载有关。     

挤压伤大鼠血清对心肌细胞蛋白质合成的影响

目的：观察挤压伤大鼠血清对培养的心肌细胞蛋白质合成的作用。方法：培养1-3 d龄SD乳鼠心肌细胞，观察挤压伤大鼠血清和正常大鼠血清对心肌细胞表面积、蛋白质含量、[³H]-Leu掺入的影响。结果：挤压伤大鼠血清组心肌细胞表面积、蛋白质含量、[³H]-Leu掺入明显大于正常大鼠血清组，且效应呈浓度依赖性。结论：肢体挤压伤大鼠血清可使培养的心肌细胞蛋白质合成增加，引起细胞肥大.     

次声作用后血浆NO、ET-1、SOD、MDA水平的变化

目的：测定8 Hz、130 dB次声不同时间暴露后大鼠血浆一氧化氮（NO）、内皮素（ET-1）、SOD、MDA水平的变化。方法：用8 Hz、130 dB的次声连续作用大鼠1、7、14、21和28 d，每天2 h，测定大鼠血浆NO、ET-1、SOD、MDA水平。结果：在暴露期间，7、14 d时大鼠血浆NO含量显著最低（P＜0.01），1 d、21 d和28 d时正常（P＞0.05）；大鼠血浆ET-1含量均明显升高（P＜0.01），7 d时升高最多，14 d时升高最少；大鼠血浆SOD活性明显降低（P＜0.01）；大鼠血浆MDA水平明显升高（P＜0.01）。结论：次声可引起大鼠血浆NO、ET-1、SOD、MDA水平的变化，发生的改变与次声暴露时间有关。     

早期胚胎心肌细胞兴奋收缩耦联模式

目的　研究胚胎心肌细胞兴奋收缩耦联的机理；方法　使用膜片钳与钙离子浓度分析系统测量酶消化法得到的小鼠胚胎心肌细胞的膜电位与细胞内钙离子浓度；结果　细胚胎心肌细胞存在两种兴奋收缩耦联模式，一种与正常成熟心肌细胞的兴奋收缩耦联模式类似，与细胞膜上的钠通道、L-钙离子通道相关；另一种由细胞内钙振荡诱发，这种钙振荡通过细胞膜上的钠钙交换蛋白引起了细胞膜电位的小幅度变化，该模式是一种更基本的兴奋收缩模式。近似熵分析表明，与后一种模式相比较，前一种模式的规律性更强。结论　胚胎心肌细胞存在两种兴奋收缩耦联模式。     

海洛因成瘾模型大鼠心肌细胞的动作电位和L型钙通道电流

背景：钙离子通道异常可导致心肌受损,海洛因可直接作用钙离子通道,从而改变心肌结构。 目的：观察海洛因成瘾致大鼠心律失常后心肌细胞超微结构、L型钙离子通道电流及心肌细胞动作电位的变化情况。 方法：SD大鼠随机分为对照组和模型组,模型组大鼠以海洛因初使剂量5 mg/(kg•d),采用逐日剂量递增法[递增剂量2.5 mg/(kg•d)]复制海洛因成瘾大鼠模型;20 d海洛因成瘾模型成功建立,继续递增剂量至第30天,进一步建立海洛因成瘾大鼠心律失常模型。 结果与结论：与对照组相比,海洛因成瘾大鼠心肌电镜结构改变主要表现在细胞核膜皱缩、核浓缩、变小,染色质集结成块,线粒体嵴排列紊乱、消失,肌小节排列紊乱、灶性断裂,肌丝辨识不清等,心肌细胞L型钙通道电流-电压曲线呈现上移趋势,90%去极化动作电位显著缩短。表明海洛因可直接致心肌结构发生病理改变,钙通道电流发生改变是造成心肌损伤的主要原因之一。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

参麦注射液改善急性心肌梗死大鼠心功能与左室重构的机制初探

目的：观察了参脉注射液对心肌梗死后心功能和心室重构的影响，及其对心肌细胞凋亡的影响。方法：（1）结扎大鼠左冠状动脉前降支建立大鼠心肌梗死心室重构模型，随机分为心肌梗死后1、2周模型组，②参麦注射液治疗1、2周治疗组，另设两假手术组。多导生理记录仪测量：左室收缩压（LVSP）、左室舒张末压（LVEDP）、左室内压最大上升速率（＋dp／dtmax）和下降速率（-dp／dtmax）以反映左室收缩与舒张功能。测定心脏心脏总重量、左室截面直径，并计算心室重量指数；HE染色、Masson染色、透射电镜观察心结构改变。（2）乳鼠心肌细胞原代培养，AngⅡ诱导凋亡模型。利用荧光染色观察凋亡形态变化；流式细胞仪检测细胞凋亡、心肌细胞线粒体膜电位；激光共聚焦显微镜检测钙离子荧光强度。免疫组化染色检测Bcl-2／Bax、Caspase-3蛋白的表达。     

参麦注射液对急性缺氧-复氧心肌细胞凋亡的影响

目的：观察参麦注射液对急性缺氧-复氧后心肌细胞凋亡的影响，并探讨其可能机制。 方法： 采用原代培养的大鼠心肌细胞，通过化学缺氧法使细胞缺氧5 min，再恢复氧供应15 min，复制心肌细胞缺氧-复氧（anoxia-reoxygenation, A/R）模型。Annexin V-FITC/PI双染色流式细胞仪检测细胞凋亡百分率；Fluo-3负载激光扫描共聚焦显微镜观察细胞内钙离子水平。 结果： A/R组心肌细胞凋亡百分率明显高于正常组，细胞内平均钙离子荧光强度也显著强于正常组（P<0.01）。参麦注射液组细胞凋亡率显著小于A/R组，同时细胞内钙超载也明显轻于A/R组（P<0.01）。 结论： 参麦注射液能有效抑制缺氧-复氧心肌细胞的凋亡，这种保护作用的机制之一可能是通过减轻细胞内Ca2+超负荷实现的。     

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.