Cell death in ischemic,reperfused porcine hearts: A histochemical and functional study

Summary The temporal development of infarcts was histochemically and functionally determined in porcine hearts. In one series of experiments (22 pigs), the distal third of the left anterior descending coronary artery (LAD) was transiently occluded for periods between 20 and 90 min and was reperfused for another 24h. At the end of the experiments, the infarcted myocardium of four tissue slices was determined with a tetrazolium stain and related to the risk region which was delineated by a fluorescent dye. Infarcts started to develop in the ischemic septum and the subendocardial layer of the free anterior wall between 20 and 35 min of ischemia. Thereafter, infarctions progressed rapidly from the inner towards the outer layer at risk. The jeopardized anterior left ventricular wall became almost completely infarcted within 60 min of ischemia. In a second series of experiments (10 pigs) recovery of systolic shortening was studied with implanted ultrasonic crystals over 3 weeks of reperfusion. At the end of the experiments, systolic shortening was about 75% of baseline level when ischemia had lasted between 20 and 35 min. Almost no recovery was observed when the occlusion time lasted 45 to 60 min. This study suggests that the assessment of myocardial infarction with a tetrazolium stain after 24 h of reperfusion corresponds very well with functional recovery after 3 weeks of reperfusion. Furthermore, determination of regional myocardial function of the ischemic, reperfused segment in the chronic stage may be considered an additional tool to evaluate therapeutic effects on infarct size in this model.The study was supported by a grant of Deutsche Forschungsgemeinschaft (DFG) Sonderforschungsbereich 330 Organprotektion Göttingen.This paper contains parts of the Habilitationsschrift of Dr. H. H. Klein.     

Establishment of a human glioma cell line bearing a homogeneously staining chromosomal region and releasing α- and β-type transforming growth factors

Summary A human glioma cell line (YKG1), which was positively identified for glial fibrillary acidic (GFA) and S-100 proteins, was established from a surgical specimen of a patient with glioblastoma. Chromosome analysis of the cells revealed a homogeneously staining region (HSR) on a marker chromosome. The assay for transforming growth factors (TGFs) in the conditioned medium of the cell line revealed that it contained high levels of - and -type TGFs, which might regulate the growth of glioblastoma and influence on the peritumoral tissues.     

恶性肿瘤病人的耳廓视诊、电探测、染色的变化

采用耳廓视诊、耳廓电探测、耳廓染色三种手段,对116例恶性肿瘤病人、120例良性疾病病人和115例健康人进行对照检查证明：恶性肿瘤病人在耳廓上的相应部位软骨增生,耳轮色素沉着,Y_2软骨增生,耳廓上代表癌区的部位及肿瘤在耳廓上的相应部位电流增大;染色时,耳轮、Y_1～Y_7（注）及耳廓相应部位着色。而一般疾病组除耳廓相应部位电流增大,染色着色及充血、血疹外,耳廓上癌区基本上无阳性反应。健康人组通过三种手段检查,基本上无阳性反应。经统计学处理有极显著差异（P＜0．01）。双盲法验证,准确率达92.65％。     

细胞内抗原免疫金银染色技术的改进

为解决细胞内抗原应用免疫金银法染色时背景过重的问题,建立了甘氨酸二次阻断的处理方法,效果较好。     

细胞因子Midkine融合蛋白的表达及其单克隆抗体的制备与应用

目的:克隆细胞因子midkine(MK)基因,表达其融合蛋白,制备抗MK单克隆抗体(mAb)并检测MK在肿瘤细胞中的表达。方法:利用MK基因的特异性引物,通过RT-PCR从人肾癌组织mRNA中扩增人MKcDNA分子。将其定向克隆于原核表达载体中,在大肠杆菌中表达MS2-MK融合蛋白。以纯化的融合蛋白免疫BALB/c小鼠,用传统的杂交瘤技术,进行细胞融合、筛选、克隆化并制备mAb腹水。用ELISA法分析其Ig亚类,用免疫细胞化学染色法检测MK在肿瘤细胞中的表达。结果:成功地克隆出MK基因并表达了MS2-MK融合蛋白。通过免疫和筛选,获得2株分泌小鼠抗人MKmAb的杂交瘤细胞,其分泌的mAb的Ig亚类分别为IgG1和IgG2a。免疫细胞化学染色显示,2株mAb与人胃癌细胞株MGC803和胃癌组织呈强阳性反应。结论:在原核细胞中获得MS2-MK融合蛋白的表达,并制备出抗MK的mAb,为研究MK的生物学活性提供了条件。     

pcDNA3.1/hIL-18真核表达载体的构建及表达

目的 :构建重组真核表达质粒pcDNA3.1/IL 18,并在哺乳动物细胞COS 7和Rlc310中进行瞬时和稳定性表达。方法 :从含hIL 18基因的中介载体 pGEM TEasy( pGEM T/hIL 18)中 ,以限制性内切酶酶切方法获得目的片段 ,克隆入真核表达质粒 pcDNA3.1( )中。以脂质体法转染COS 7和Rlc310细胞 ,用RT PCR检测IL 18mRNA的水平 ,免疫组化染色法检测蛋白表达。结果 :构建了hIL 18基因的重组真核表达质粒pcDNA3.1/IL 18,并可在哺乳动物细胞中瞬时、稳定表达 ,获得了可稳定表达hIL 18基因的Rlc310细胞株。结论 :pcDNA3.1/IL 18的构建及表达 ,为IL 18抗肿瘤作用的研究奠定了基础     

免疫电镜技术运用过程中电子染色方法的选用

目的探究免疫电镜不同染色方法对免疫组化阳性实验结果影响的关系。方法组织切片经常规免疫组化（雌激素受体GPR30）并行DAB-硫酸镍铵显色后进行电镜切片，然后分为双氧铀-柠檬酸铅双染、双氧铀单染与未染色3组，以便对不同电子染色结果进行比较。结果GPR30免疫阳性产物位于细胞核外的膜性结构上，在铀-铅双染组显示很高的电子密度，但是背景染色也很深，在铀单染组的反差比较好，而未染色组的反差更好。结论免疫电镜技术中针对不同的免疫阳性反应选用不同的电子染色方法，有利于阳性结果的判断与鉴别。     

改良单宁酸-氯化铁法媒染子宫血管的实验研究

目的：光镜下观察大鼠子宫各部位血管构筑。方法：应用单宁酸一氯化铁法(TA—Fe法)灌流固定大鼠，取子宫不同位置做冰冻切片，氯化铁显色，常规脱水、透明、封片，显微摄影。结果：子宫壁出现类似电镜负染色效果，组织结构灰黑色，血管双线条状，分支明显，过管壁切面可见内皮细胞或平滑肌；子宫动脉进入肌层分支并形成一、二级血管网和多支环状动脉，环行动脉或二级血管网发出的微动脉和毛细血管进入粘膜，形成密集的三级毛细血管网。结论：经灌流的子宫标本较长时间保存在媒染固定液内，导致血管外结构也被媒染；子宫壁有三级血管网；每个子宫有多支环状动脉；各段子宫血管构筑不尽相同。     

Secretin and VIP-stimulated adenylate cyclase from rat heart

The exact positions of microelectrodes used to measure thePO₂ in the cerebral cortex of the rat were determined by staining the tissue with Alcian Blue. The measurement sites were subsequently located under a light microscope and correlated with the capillary and cellular arrangement of the cortex. The microelectrodes used for thePO₂ measurements were made of gold glass fibers; the Alcian Blue was injected hydrostatically through a micropipette attached to thePO₂ microelectrode. The sites where dye had been deposited were seen under a light microscope as green blue spots about 100 m in diameter. The capillaries were visualized by silver nitrate perfusion. Differences between the localPO₂ values in the neo- and the archeocortex were found. In the neocortex the meanPO₂ was 31 mm Hg, capillary volume 1.6%, capillary surface area 980/mm², capillary length 13.5/mm; whereas in the archeocortex these values where 21 mm Hg, 0.9%, 820/mm² and 9.4/mm respectively. These data indicate a relationship between the microcirculatory transport system and the local oxygen tension and provide further evidence that the meanPO₂ level tends to decrease when moving from the surface into the archeocortex.Supported by the Deutsche ForschungsgemeinschaftReported in part at the 3rd Symposium of ISOTT, Cambridge, GB, 1977; and at the 27th International Congress of Physiological Sciences, Paris, France, 1977     

Characterization of a monoclonal antibody (BG3C8) that reacts with basal cells of stratified epithelia

Monoclonal antibodies were produced against a suspension of formaldehyde fixed human epidermal cells. The supernatant fluid of one clone (BG3C8) yielded a bright immunofluorescent staining of basal cells both in cryostat sections of human split skin and in preparations of purified basal cells. As determined by one- and two-dimensional gel immunoblotting of epidermal basal cell proteins the antibody recognized a minor basic polypeptide of 55,000 apparent molecular weight that was not present in extracts of cultured cell lines of epithelial, fibroblast and lymphoid origin. The distribution of the 55,000 molecular weight protein in normal human tissue was determined by immunohistological staining of cryostat tissue sections that included: central nervous, endocrine, female and male reproductive, alimentary, lymphatic-haemopoietic, respiratory and urinary systems, skin and its appendages, mesenchymal tissue (bone, cartilage, muscle, connective tissue, blood vessels, nerves and synovia) as well as placenta and umbilical cord. The results showed a restricted distribution of this antigen which was found only in basal cells of most stratified or pseudostratified epithelia and in myoepithelial cells. This antibody may be useful in the study of normal and pathological differentiation in various epithelial disorders.