心肌梗死大鼠模型的建立及疾病进程评价

背景:心肌梗死是冠心病最严重的一种临床类型,抗心肌缺血类药物的研发已成为研究热点。建立合适的动物模型并探讨其病理生理机制,可为新药研发过程中药物的药效学评价提供有效的工具,促进心血管疾病有效治疗药物的开发。目的:评价急性心肌梗死损伤后大鼠的心脏功能、组织形态和心肌细胞超微结构改变。方法:30只雄性SD大鼠随机分为假手术组及模型组,每组15只。模型组采用冠状动脉左前降支结扎法制备心肌梗死模型,假手术组只穿线不结扎。分别在造模后第7,14,28天,心电图观测大鼠Ⅱ导联ST段改变情况;开胸观察心脏大体改变情况;2,3,5-三苯基氯化四氮唑染色法测定心肌梗死面积;苏木精-伊红染色法观察心脏组织病理改变;Masson染色法观察心肌组织纤维化程度;透射电镜观察心肌细胞超微结构改变。结果与结论:(1)与假手术组比较,模型组大鼠术后7 d心电图可见J点明显抬高,14 d可见Q波的波形增宽、振幅增大;(2)心脏大体观察可见,模型组大鼠术后7,14 d时左心室肥大,28 d时心脏萎缩;(3)2,3,5-三苯基氯化四氮唑染色显示,模型组大鼠心梗面积进展性增加;(4)苏木精-伊红和Masson染色显示,心肌细...     

大鼠急性心肌梗死区域miRNA-29和miRNA-21的动态变化与心功能的关系

目的 探讨大鼠急性心肌梗死后早期梗死区域miRNA-29和miRNA-21的动态变化与心功能的关系.方法 SD雄性大鼠25只,结扎冠状动脉前降支,建立大鼠心肌梗死模型,分为术后7、14、21、28 d组和假手术组(对照组),每组各5只.彩色超声心动图检测大鼠左室射血分数(LVEF)、左室短轴缩短率(LVFS)和左室舒张末期内径(LVIDd).石蜡切片,苏木精-伊红(HE)和Masson染色.Western blotting检测大鼠心肌梗死后梗死区的胶原纤维I(I)和弹性蛋白(E)的变化.实时荧光定量聚合酶链法检测大鼠心肌梗死区域miRNA-29和miRNA-21的水平.结果 大鼠心肌梗死后,心功能下降,2周时LVEF和LVFS下降至最低,后缓慢升高(P<0.05);胶原I逐渐升高(P<0.01),弹性蛋白E逐渐降低(P<0.05).而且较对照组,心肌梗死区miRNA-29于第1周上升明显,第2周迅速下降(P<0.01);miRNA-21第2周明显上升,后逐渐下降(P<0.01).结论 miRNA-29和miRNA-21表达升高可能在急性心肌梗死早期的心功能降低中起重要作用.     

心梗后心衰大鼠心肌细胞凋亡时相变化特点观察

目的 :观察分析心梗后心衰大鼠心肌细胞凋亡及相应调控基因蛋白表达的时相变化特点。方法 :以左冠状动脉穿线结扎的方法造成大鼠急性心梗 ,经阻抗法评价心功能 ,制成心衰动物模型 ,持续喂养 8周。并设假手术组 (穿线不结扎 )作为对照。在此过程中 ,选取 3ｄ、5ｄ、1 0ｄ、4周、8周 5个时间点 ,分别以原位末端标记法 (ＴＵＮＥＬ)检测模型大鼠梗塞边缘区 (梗塞和非梗塞区交界处 )、健存区和假手术组心肌组织中的凋亡细胞 ,Ｓ -Ｐ免疫组化法检测心肌细胞中的ｂｃｌ- 2、Ｂａｘ基因蛋白表达的变化。结果 :模型组边缘区各个时间点之间比较 ,早期…     

EMMPRIN在大鼠肾间质纤维化过程中的表达变化

目的 检测细胞外基质金属蛋白酶诱导因子(EMMPRIN)在大鼠肾间质纤维化模型中的表达变化.方法 64只Sprague-Dawley大鼠随机分为假手术组(Sham组)和模型组(UUO组),每组16只.假手术组,仅游离输尿管不结扎,模型组,行单侧输尿管结扎术,分别于术后3、7、14 d处死大鼠并留取肾脏组织.肾脏标本行HE、Masson染色方法检测间质纤维化损伤的程度.应用免疫组织化学方法检测肾组织中EMMPRIN,α-SMA,E-cadherin的表达.结果 HE染色显示:UUO术后肾间质肾小管萎缩、扩张,胞外基质沉积,炎细胞浸润增加.Masson染色显示:UUO术后大鼠肾组织胶原纤维组织增生,出现明显的间质纤维化改变.免疫组织化学结果显示:与假手术组相比,UUO模型3d组的EMMPRIN表达显著增高,同时有α-SMA表达升高和E-cadherin表达显著下降(P＜0.05);随着梗阻时间的延长,模型7d组EMMPRIN的表达逐渐减弱(P＜0.05).结论 EMMPRIN可能参与了UUO大鼠肾间质纤维化的早期上皮间质转分化过程,而随着纤维化程度加重,后期表达逐渐减少.     

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。     

有氧运动协同骨髓干细胞动员对心肌梗死后心电图和血流动力学指标的影响

背景：心电图和血流动力学是评价心功能康复的有效指标,目前已经证实有氧运动或骨髓干细胞动员单一因素干预均可对心肌梗死动物心电图和血流动力学产生良好影响,而二者联合干预对心电图和血流动力学指标的影响尚未见文献报道。 目的：探讨有氧运动联合骨髓干细胞动员对缺血心脏心电图和血流动力学部分指标的影响。 方法：结扎大鼠左冠状动脉前降支制作急性心肌梗死模型,心肌梗死运动组和心肌梗死运动动员剂组大鼠于造模后1周在电动跑台进行有氧运动训练,每周训练5 d,持续8周。心肌梗死动员剂组和心肌梗死运动动员剂组大鼠在造模后3 h皮下注射生理盐水稀释的重组人粒细胞集落刺激因子10 μg/(kg•d),连续使用5 d。8周后检测心电图和血流动力学部分指标评价心功能。 结果与结论：心肌梗死大鼠左心室收缩压、左室内压最大上升速率和左室内压最大下降速率值均明显降低,左室舒张末压升高,提示心梗后心脏已发生心功能不全;心肌梗死运动组和心肌梗死动员剂组大鼠左心室收缩压、左室内压最大上升速率和左室内压最大下降速率值均有一定程度的升高,左室舒张末压有所下降,提示有氧运动和骨髓干细胞动员均能改善心梗大鼠心肌收缩和舒张功能;心肌梗死运动动员剂组大鼠心功能的各项评价指标更接近于正常对照组大鼠,说明有氧运动协同骨髓干细胞动员显著增强了大鼠心肌收缩性能,使心肌收缩/舒张功能都得到显著改善。     

bFGF通过促进心脏血管新生改善心肌梗死后小鼠心脏重构

目的:探讨碱性成纤维细胞生长因子(basic fibroblast growth factor,b FGF)对心肌梗死小鼠心脏的保护作用及机制。方法:采用异氟烷麻醉C57/B6小鼠(8~12周龄)后,侧开胸结扎冠状动脉左前降支,建立小鼠心肌梗死模型;设立假手术组为对照,心肌梗死模型小鼠随机分为心梗组和bFGF给药组,其中bFGF组小鼠心梗7d后给予5μg b FGF隔天腹腔注射给药;在小鼠心梗第28天时采用心脏多普勒超声检测心功能,以左室舒张末期内径、左室收缩末期内径、左心室射血分数和左心室短轴缩短分数评价心脏功能改变;28 d后处死小鼠,行病理切片观察心肌纤维化程度和心肌梗死区内血管新生的情况;Western blot检测血管新生指标。结果:bFGF给药组小鼠心肌纤维化程度较心梗模型组明显减少;第28天行超声心动图检查结果示,心梗组小鼠心功能较假手术组差,而bFGF给药组小鼠心功能与心梗组比较有明显的改善;小鼠心肌病理切片免疫荧光观察结果发现,bFGF给药组心肌梗死区的新生血管比心梗组明显增多;Western blot实验表明,bFGF能够激活AKT/HIF-1α/VEGF通路。结论:隔天腹腔注射bFGF能够减少心肌梗死小鼠心肌纤维化,改善心功能。bFGF可能通过AKT/HIF-1α/VEGF信号通路促进血管新生,从而保护心脏。     

心肌梗死后心力衰竭小鼠心肌组织内质网应激相关凋亡途径的研究

目的:探讨心肌梗死后心力衰竭小鼠心肌组织内质网应激介导的凋亡途径。方法:结扎小鼠左冠状动脉前降支建立心肌梗死后心力衰竭(心衰)模型,32只小鼠利用随机数字法分4组:假手术组、心肌梗死后2周组、4周组和6周组,采用超声心动图检查观察心室扩张及心功能变化情况,Western blotting检测内质网分子伴侣GRP78蛋白以及内质网应激相关凋亡蛋白CCAAT/增强子结合蛋白ε(CCAAT/enhancer-binding proteinε,CHOP)、c-Jun氨基端激酶(c-Jun N-terminal kinase,JNK)及其磷酸化水平、caspase-12及其活性剪切片段的表达。采用TUNEL法观察心肌细胞的凋亡情况。结果:与假手术组相比较,心肌梗死后心力衰竭的小鼠左心室扩大,心功能下降。小鼠心肌组织GRP78、CHOP蛋白表达增高(P0.05)。JNK、caspase-12蛋白表达没有明显变化,但其活化形式磷酸化JNK及剪切后的caspase-12表达增高(P0.05)。TUNEL染色显示心肌梗死后心力衰竭的小鼠心肌组织凋亡明显增多(P0.05)。结论:心肌梗死后心力衰竭诱导内质网应激反应,并伴随着其相关的CHOP、JNK、caspase-12三条通路的激活,提示内质网应激可能参与了心梗后心衰中心肌细胞的凋亡。     

前列腺素E1通过抑制内质网应激保护心肌梗死后大鼠的心脏

目的:探讨前列腺素E1(PGE1)对心肌梗死(MI)后大鼠心脏的影响及相关分子机制。方法:将SPF级雄性SD大鼠分为假手术组、模型组和模型+PGE1组。通过结扎冠状动脉左前降支建立MI大鼠模型。通过超声心动图分析大鼠心功能变化;通过HE和Masson染色分析心肌组织形态学变化;通过TTC染色评估心肌梗死情况;通过TUNEL法检测心肌细胞死亡情况;通过免疫组化和Western blot检测内质网应激(ERS)相关因子葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)和caspase-12,以及凋亡相关因子Bcl-2、Bax和cleaved caspase-3的表达情况。结果:与假手术组比较,模型组大鼠心功能降低,心肌组织出现病理形态学改变,心肌梗死面积扩大,心肌细胞死亡增多,心肌组织中GRP78、CHOP、caspase-12、Bax和cleaved caspase-3的蛋白水平升高,Bcl-2表达降低(P<0. 01)。与模型组比较,模型+PGE1组大鼠心功能明显提高,心肌组织病理损伤明显减轻,心肌梗死的面积显著减小,心肌细胞死亡显著减少,心肌组织中GRP78、CH...     

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。     

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.     

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 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.