结晶紫染色法显示弹力纤维的应用

病理诊断常用 Comori醛复红 ,Weigert来复红 ,Verhochff法 Unna地衣红对弹力纤维[1] 进行特殊染色来观察显示皮肤、肺、血管组织中弹力纤维的增生、变性、散乱、断裂 ,其染色结果可呈紫色、兰黑色、褐色 ,但复染苏木素定位胞核和 VG显示胶原纤维染色后 ,对比不鲜明 ,经尝试用结晶紫改良 Weigert染色法后 ,弹力纤维呈绿色 ,胶原纤维红色 ,肌肉组织及背景黄色 ,对比鲜明 ,色彩艳丽、清晰 ,收到了理想效果。材料和方法1 .材料1 .1　标本来源 :本室活检和尸检正常或病变的心肌、皮肤、肺组织 ,经福尔马林固定 ,常规石蜡切片 5 μm。1 .2　染…     

小鼠心脏发育的形态学研究

目的建立小鼠心脏正常发育的时间表以及对应的形态学特征模式.方法小鼠胚胎ED8.5、ED9.5、ED10.5、ED11.5、ED12.5、ED14.5、ED16.5、ED18.5和P1(postnatal day)(出生后1 d的仔鼠)标本,进行整体或心脏部位不同轴向切片,HE染色,采用PCTV图像分析系统,对各时相小鼠心脏形态发育特征进行研究.结果 ⑴细胞结构发育的时空模式:① ED8.5时,生心板形成;ED9.5时,心肌细胞呈不规则的纺锤形,细胞的大小多样化,细胞核小;ED10.5时,小血管和着色较浅的肌原纤维出现,细胞之间连接较松;ED11.5时,心肌纤维排列较紧,纵断面上呈细长形,横断面上呈不规则多角形;ED12.5时,细胞核着色更清晰,心肌细胞形状逐渐规则,细胞之间紧密连接,同时闰盘结构出现在心室心肌细胞.②ED12.5时心肌小梁结构第一次在心室出现,ED14.5时增厚,而在心房少见心肌小梁.⑵心室结构的形成和心脏发育的成熟:①心肌间充质网络结构在ED10.5的心室中明显呈现,随着它的发育,心室的心内膜在ED11.5出现,心室心外膜可以辨认.②房室隔在ED12.5完全形成,心内膜垫在ED12.5开始发生并快速发育,促进室间隔在ED14.5完全形成.③心包膜在ED16.5可明显辨认,心包膜腔形成,此时近段流出道心内膜垫完全心肌细胞替换.结论肌原纤维细胞和心肌间充质细胞同时在ED10.5出现,提示肌原纤维对心肌细胞的成型和心肌化起作用.细胞的结构变化和心肌层的成熟过程,显示小鼠心脏部位成熟时间的不同,心室成熟相对较晚.     

心室肌的大体与显微的层次解剖研究

目的:通过对心室肌的大体解剖与组织学切片显示心室肌纤维的层次走向,比较心肌传统概念与心肌带理论的差异.方法:(1)应用大体解剖、组织切片HE染色、虚拟切片扫描方法显示心室肌的纤维走向及层次构造.(2)将防腐固定后的成人正常心脏顺心肌纤维方向分层解剖心肌,使之展开成带,验证心肌带理论.结果:将防腐固定的成人正常心脏经改良的解剖方法解剖,可解剖出一条完整的心肌带,分为2个环,4个段.心肌带呈螺旋走向.用大体解剖与组织切片HE染色、虚拟切片扫描显示右心室肌起始于纤维三角为一条单层肌肉带,左心室肌分为三层肌纤维起始于纤维三角与纤维环,肌纤维从纤维三角与纤维环持续由外向内转变呈现为一种交错结构,纵行的浅层心肌纤维在心尖以90°的角返回心底方向,构成深层心室肌形成肉柱,部分心肌纤维斜、横行构成中层心室肌.三者在心尖呈现出一种螺旋形结构.应用虚拟切片扫描技术能完整、清楚地显示心室肌某一切面的结构走向.结论:心肌传统理论与心肌带理论并未完全冲突,心肌带理论的降段、左室断、升段与传统理论的浅、中、深三层相对应.     

痛风的临床病理特征分析并文献复习

目的:总结痛风临床病理特点。方法:回顾性分析1例痛风患者的生化机制、临床病理特征、刚果红染色、PAS染色特点、鉴别诊断要点,并复习相关文献。结果:患者主要临床表现为间断性多关节肿痛3年,加重伴发热3个月。体格检查发现患者有多发性皮下结节、多关节肿胀压痛。左腕、左肘皮下结节活检,经HE染色后光镜查见大量肉芽肿性病变,有的多核巨细胞内查见被吞噬的异物,有的病灶尚查见呈均质状物(尿酸盐结晶),其周围有较多异物巨细胞及纤维结缔组织包绕呈结节状,在结节的周边纤维血管周围可查见残留分化成熟的淋巴细胞及少数嗜酸性粒细胞。刚果红、PAS染色均为阴性。偏光显微镜下,刚果红未查见绿色强折光晶体,但见多量略呈淡黄色具有强折光性的晶体呈棒状或梭形。结论:痛风在刚果红染色偏光显微镜下观察呈淡黄色梭形或针状结晶,具有强折光性晶体,但这是否是痛风在刚果红染色的特征尚有待于进一步研究。     

慢性心力衰竭大鼠心肌毛细血管密度及血管内皮生长因子变化

目的观察慢性心力衰竭大鼠心肌毛细血管密度及血管内皮生长因子(VEGF)变化,探究冠脉微循环障碍的病理特点及病因机制.方法实验组(n=15)皮下注射异丙肾上腺素,对照组(n=10)皮下注射生理盐水,间隔24h,连续2次.12周后测定血液动力学;计算左心室重量/体重;HE染色、Masson染色分别观察左心室病理改变、胶原变化;西非单叶豆素组织化学染色结合图像分析确定心内膜下心肌毛细血管密度、心肌细胞密度、毛细血管密度与心肌细胞密度的比值(毛细血管/心肌细胞);观察心内膜下心肌VEGF免疫组织化学变化.结果同对照组比较,实验组左心室收缩、舒张功能下降(P<0.05);左心室重量/体重升高(P<0.001);心内膜下心肌散在坏死,胶原沉积;心内膜下心肌毛细血管密度、心肌细胞密度、毛细血管/心肌细胞下降(P<0.05);VEGF合成增加(P<0.001).结论慢性心力衰竭大鼠心内膜下心肌毛细血管分布稀疏;该区域毛细血管代偿性生成减少与心肌VEGF表达无关.     

心肌组织特异性Isca1敲除造成新生大鼠心脏结构异常

目的通过对心肌组织特异性Isca1敲除大鼠进行磁共振分析及病理学分析,探究心肌特异性敲除Isca1对大鼠心脏结构影响。方法繁育心肌组织特异性Isca1敲除大鼠,PCR技术鉴定大鼠基因型及基因敲除效率,对新出生0.5d及2.5d心肌组织特异性Isca1敲除大鼠进行核磁共振影像分析,对新出生2.5d心肌组织特异性Isca1敲除大鼠心肌组织进行H&E染色及透射电镜分析。结果心肌组织特异性Isca1敲除大鼠敲除效率大于78%;与野生型相比,0.5d心肌组织特异性Isca1敲除大鼠心脏未见显著扩张;2.5d心肌组织特异性Isca1敲除大鼠心脏右室呈扩张趋势;2.5d心肌组织特异性Isca1敲除大鼠肌纤维排列不齐,出现排列紊乱,无层次或极向,部分心肌纤维出现溶解断裂,肌节和Z线模糊,出现肌膜损伤,线粒体嵴断裂,肿胀明显。结论心肌组织特异性Isca1敲除造成新生大鼠心脏结构异常。     

心内膜下心肌梗死的常见误诊病例浅析

目的:探讨心内膜下心肌梗死的常见误诊原因及对策。方法:回顾性分析9例心内膜下心肌梗死患者的病历资料,包括病史、系统体格检查,ECG、心肌酶谱、胸片、超声心动图检查等。结果:9例心内膜下心肌梗死患者均有临床症状、ECG及心肌酶学指标的动态变化,诊断明确,以冠状动脉病变为其主要病因,在此基础上由其他因素诱导发病。结论:临床医师对心内膜下心肌梗死认识不足、对ECG和心肌酶谱的特异性动态改变未充分认识,是导致误诊的主要原因,建议详细询问病史、系统体格检查及辅助检查,综合判断明确诊断。     

激光心肌血管重建术后激光管道的形态学观察

为临床研究和应用激光心肌血管重建术提供解剖学基础,本文在１６只杂交犬的心肌梗塞模型上,行ＣＯ２激光心肌打孔。术后３或６个月取出心脏,观察不同时间未铸型及铸型激光心肌管道的形态特征。结果表明激光管道心内膜侧管径大于心外膜侧,管道多呈锥形。     

宫颈特殊染色技术对子宫颈癌及宫颈上皮内瘤变初筛的临床研究

目的:探讨宫颈特殊染色技术筛查方法对宫颈癌及癌前病变的筛查意义。方法:本研究通过对1963例就诊我院妇科门诊的患者进行宫颈特殊染色检查(FRD),以组织病理学检查结果为标准,分析FRD宫颈特殊染色的临床意义。结果:1963例患者行宫颈特殊染色检查及对初筛阳性患者行阴道镜下活检,根据活检病理结果进行分析,CINI阳性率80.77%℅,CINII81.25%,CINIII100%℅,侵润癌100%,总阳性率90.50%。结论:利用亚甲蓝显色和醋酸白化反应双重定位及指示,不仅可提高宫颈癌及癌前病变的检出率,而且操作简便,判读容易,结果快速,成本低廉。     

霉菌、幽门螺杆菌及双菌种感染在胃溃疡、胃癌中检出的意义

探讨霉菌、幽门螺杆菌(Hp)单菌种感染和霉菌、Hp(双菌种)同时感染在胃癌及胃溃疡中的组织病理学变化、发病情况及意义。采用常规石蜡切片,HE染色和PAS、Giemsa特殊染色、免疫组织化学染色及PCR方法,对223例慢性浅表性胃炎、111例慢性萎缩性胃炎、116例胃溃疡、121例胃癌纤维胃镜活检标本进行回顾性研究。结果显示,慢性浅表性胃炎、慢性萎缩性胃炎未检出双菌种感染。胃溃疡双菌种感染11例,检出率9.5%;胃癌双菌种感染21例,检出率17.4%。双菌种感染在胃癌及胃溃疡中的发现,表明双菌种感染可能是导致胃溃疡、胃癌发生的又一致病因素。     

Elastin distribution in the normal uterus, uterine leiomyomas, adenomyosis and adenomyomas: a comparison

OBJECTIVE: To describe the histologic distribution of elastin in the nonpregnant human uterus, uterine leiomyomas, adenomyosis and adenomyomas. STUDY DESIGN: Uteri were obtained from women undergoing hysterectomy for benign conditions, including 26 cases of uterine leiomyomas, 24 cases of adenomyosis, 18 adenomyomas and 6 cases of autopsy specimens. Specific histochemical staining techniques were employed in order to demonstrate the distribution of elastin. RESULTS: The distribution of elastin components in the uterus was markedly uneven and showed a decreasing gradient from outer to inner myometrium. No elastin was present within leiomyomas, adenomyomas or adenomyosis. CONCLUSION: The distribution of elastin may help explain the normal function of the myometrium in labor. It implies that the uneven distribution of elastin components and absence of elastin within leiomyomas, adenomyomas and adenomyosis could be of some clinical significance. The altered elastin distribution in disease states may help explain such symptoms as dysmenorrhea in uterine endometriosis.     

蚯蚓纤溶酶底物特异性

蚯蚓纤溶酶是具有较强溶栓作用的丝氨酸蛋白酶,为研究蚯蚓纤溶酶在溶栓的同时是否对其他血液蛋白有破坏作用,本实验以一些血液功能蛋白为底物,用蚯蚓纤溶酶在体外对其进行水解,采用SDS—PAGE检测水解前后底物成分的变化。结果显示,蚯蚓纤溶酶短时间内能完全水解纤维蛋白和纤维蛋白原,长时间作用下能部分水解牛血清白蛋白和人免疫球蛋白重链,不水解牛血红蛋白、牛血超氧化物歧化酶和牛凝血酶原。表明蚯蚓纤溶酶对血栓成分具有较强的识别和水解能力,而对其他血液蛋白作用微弱,说明蚯蚓纤溶酶作为药物使用时具有较强的溶栓和预防血栓形成的作用,且副作用较小。     

Evolution of the elastic fiber network of the human uterine cervix before, during and after pregnancy. A quantitative evaluation by automated image analysis

To assess the evolution of the elastic fiber network of the human uterine cervix before, during and after pregnancy, biopsy samples were obtained from 49 women. The high affinity of the polyphenolic compound (+)-catechin for elastin was used to stain the elastic fibers selectively, and enabled automated image analysis. In the human uterine cervix, the elastic fiber network is made up of: (1) fibers running parallel to the basement lamina of the epithelium, and (2) thinner, perpendicular fibers. Quantification using automated image analysis shows a decline in the cervical elastin content from a prepregnancy level of 1.33 +/- 0.08 (SEM) to 0.73 +/- 0.09% (Vv) at the end of pregnancy. In parallel with a constant decline, dissociation and disorganization of the fibers become more clearly evident as pregnancy progresses. However, by 5-7 weeks postpartum the elastic fiber network appears almost completely restructured. These changes support a role of elastin in the processes of cervical maturation and reconstruction during pregnancy and after delivery.     

Mouse atrio-ventricular valve ultrastructure morphometrical correlations

Morphometric data on mouse atrio-ventricular valve ultrastructure are reported. The statistical analysis of the volumetric density percentage for cellular and extracellular valve components and of endothelial plasmalemmal vesicle density for the different endocardial domains (atrial, valvular and ventricular) showed: the bicuspid compared to the tricuspid valve has a more important lymphatic drainage, less vascularization, higher endothelial plasmalemmal vesicle density and more macrophages, striated muscle cells and collagen along with fewer interstitial cells, nervous terminals and elastin in the leaflet; the valvular endothelium as compared to other endocardial domains has a higher density of plasmalemmal vesicles, considering the results for both endothelial fronts (luminal and abluminal).     

Relationship between the activity of the ethylene-forming enzyme and the level of intracellular 2,4-dichlorophenoxyacetic acid in pear cell culturesin vitro

Ethylene production by auxin-dependent pear cells culturedin vitro falls rapidly when they are deprived of 2,4-D. This phenomenon is associated with a decrease in ACC production. Readdition of 2,4-D causes a resumption of ACC production and ethylene synthesis. Ethylene-forming enzyme (EFE) activity, although never limiting, decreases sharply during 2,4-D depletion and rises again upon addition of 2,4-D. This increase in the EFE activity is not a rapid response to 2,4-D, since it requires several hours. Changes in EFE activity follow the same pattern as changes in 2,4-D concentration; the decrease in EFE activity is also concomitant with a decrease in the ability of 2,4-dinitrophenol to inhibit ethylene production. The possible role of auxins in the modulation of EFE activity is discussed.     

Relationship between the activity of the ethylene-forming enzyme and the level of intracellular 2,4-dichlorophenoxyacetic acid in pear cell culturesin vitro

Ethylene production by auxin-dependent pear cells culturedin vitro falls rapidly when they are deprived of 2,4-D. This phenomenon is associated with a decrease in ACC production. Readdition of 2,4-D causes a resumption of ACC production and ethylene synthesis. Ethylene-forming enzyme (EFE) activity, although never limiting, decreases sharply during 2,4-D depletion and rises again upon addition of 2,4-D. This increase in the EFE activity is not a rapid response to 2,4-D, since it requires several hours. Changes in EFE activity follow the same pattern as changes in 2,4-D concentration; the decrease in EFE activity is also concomitant with a decrease in the ability of 2,4-dinitrophenol to inhibit ethylene production. The possible role of auxins in the modulation of EFE activity is discussed.     

Age-related changes in the content of fibrous proteins in the rat tissues.

An attempt was made to establish a relationship between the content of elastin and collagen in the rat tissues during the process of aging. The content of collagen fractions and elastin in the rat liver, lung and skin, as well as the elastolytic activity of blood serum were determined. It was found that the concentration of elastin as well as the elastolytic activity of blood serum are increasing during the maturation of rats and the total collagen content is increasing too. After the animals reached the age from twelve to twenty four months--above mentioned values began to decrease. It is concluded that the changes in the content of the two fibrous proteins of the connective tissue depend on age.     

The effect of elastin damage on the mechanics of the aortic valve

Porcine bioprosthetic heart valves degenerate and fail mechanically through a mechanism that is currently not well understood. It has been suggested that damage to the elastin component of prosthetic valve cusps could be responsible for changes in the mechanical function of the valve that would predispose it to increased damage and ultimate failure. To determine whether damage to elastin can produce the structural and mechanical changes that could initiate the process of bioprosthetic valve degeneration, we developed an elastase treatment protocol that fragments elastin and negates its mechanical contribution to the valve tissue. Valve cusps were mechanically tested before and after digestion to measure the mechanical changes resulting from elastin damage. Elastin damage produced a decrease in radial and circumferential extensibility (from 43 to 18% strain radially and 12 to 7% strain circumferentially), with a slight increase in stiffness (1.3-2.6kN/m for radial and 10.6-11.9kN/m for circumferential directions). Digestions with trypsin, which does not cleave elastin, confirmed that the changes in mechanics of the circumferential samples were likely due to the nonspecific removal of proteoglycans by elastase, while the changes in the radial samples were indeed due to elastin damage. Removing the mechanical contribution of elastin alters the mechanical behavior of the aortic valve cusp, primarily in the radial direction. This finding implies that damage to elastin will distend the cusps, reduce their extensibility, and increase their stiffness. Damage to elastin may therefore contribute to the degeneration and failure of prosthetic valves.     

Endocardial fibroelastosis in children to one year of age]

Out of 2398 autopsies of newborns and infants performed between 1976 and 1985, thirty two cases of endocardial fibroelastosis were found, i.e. 10.6% of the congenital heart diseases in this age group. Endocardial fibroelastosis was most frequent (44%) in infants aged between 3 and 6 months. Seventeen cases of the disease were seen in male infants and 15 cases in female infants. Twenty cases of the endocardial fibroelastosis (62.5%) were of isolated character (primary) whereas 12 cases (37.5%) were of the secondary character associated with other heart abnormalities. Nine cases (about 28%) coexisted with congenital abnormalities of joint and bone system and respiratory tract. A morphologic analysis of the endocardial fibroelastosis with particular reference to its etiopathogenesis suggest its congenital origin or (and) an effect of infection on myocardium. It was also suggested that endocardial fibroelastosis may be a symptom of collagenic disease. About 70% of autopsied infants were hospitalized over 3 days what meant that the course of the disease was severe. Endocardial fibroelastosis was diagnosed clinically only in 6 cases (18.75%).     

Stimulation of ACC-dependent ethylene production in citrus leaf discs by light

The influence of light and darkness incubation on in vivo ethylene forming enzyme (EFE) activity in citrus ( Citrus sinensis L. Osbeck cv. Salustiana) mature leaf discs was studied. Leaf discs incubated in light produced higher amounts of ethylene than in darkness. Transfer of discs from light to the dark resulted in a marked inhibition of EFE activity, whereas transfer of discs from the dark to light enhanced ethylene forming activity considerably. Light did not affect 1-aminocyclopropane-l-carboxylie acid (ACC) uptake. Incubation in a CO₂-eniiched atmosphere enhanced EFE activity both in light and in darkness, but light stimulation of EFE activity was apparently not affected by CO₂. Effects of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU, inhibitor of photosynthetic electron flow) and KCN (inhibitor of cytochrome oxidase) were studied. DCMU at 0.2 m M inhibited EFE activity in light, whereas no effect was detected in the dark. On the other hand 1 m M KCN stimulated EFE activity in the light, and no significant effect was observed in the dark. CoCl₂ at 1 m M inhibited ACC-dependent ethylene production, suggesting that ethylene production from ACC is mediated by EFE in citrus leaf discs both in light and in the dark. Cycloheximide also inhibited EFE activity in the light and no effects were detected in the dark. Therefore protein synthesis in light (perhaps EFE synthesis) could be required for the light stimulation of the in vivo EFE activity.