GPC-Ⅱ-3液间歇低温灌流保存兔肺的形态学观察

目的探讨对移植供肺具有长期保护作用的新技术。方法用自制的GPC-Ⅱ-3液,以间歇低温灌流的方法保存兔肺24～192h,观察其组织结构的变化。结果保存120h以内的肺,其组织结构与对照组无明显区别,肺泡、各级支气管、血管的结构清楚、支气管上皮结构完整清楚,上皮细胞胞核清晰,染色质分布均匀、肺泡完整,肺泡上皮和毛细血管内皮清楚。部分肺泡腔中,可见结构清楚的尘细胞及其吞噬的粉尘颗粒;保存144h的肺,细胞成分的染色似乎有所加深,其余结构无明显变化。保存168h的肺,肺泡、各级支气管、血管的结构依然清楚,但细胞成分的染色有所加深。部分支气管上皮细胞有脱落现象。保存192h的肺,细胞成分的染色明显加深,有固缩迹象,支气管上皮有脱落现象加重。结论用GPC-Ⅱ-3液以低温冷藏的方法能够保存兔肺的组织结构120h。     

GPC-Ⅱ-3液间歇低温灌流保存兔心脏的形态学研究

目的探讨对移植供心具有长期保护作用的新技术。方法新西兰大白兔25只随机分为实验组(20只)和对照组(5只),用自制的GPC-Ⅱ-3液,间歇低温灌流连续保存(1～4℃)兔心24～192h,观察其组织结构的变化。结果保存120h以内的兔心脏,其组织结构与对照组无明显区别;心肌细胞、心内膜、心外膜清晰;心肌细胞横纹清楚,细胞核染色质分布均匀;心肌细胞间质无明显水肿、增宽现象;心肌细胞核的双层核膜清晰;线粒体无肿胀,其双层嵴膜清楚,基质致密;肌原纤维清楚,排列整齐,明带、暗带分明;心肌细胞闰盘清楚,结构完好;心肌毛细血管内皮细胞及其基膜完整、结构清楚。保存144h的兔心,肌原纤维清楚,排列整齐,明带、暗带分明,但线粒体嵴有所减少或消失,毛细血管内皮细胞开始出现溶解现象。保存168～192h的心脏,心肌细胞核染色质有较明显的凝集现象,甚至出现小片状坏死;线粒体明显肿胀,嵴消失;毛细血管内皮细胞溶解,但其基膜仍然完整。结论用GPC-Ⅱ-3液以间歇低温灌流的方法能够保存兔心脏的组织结构120h。     

自制GPC-Ⅱ-3液对兔肝低温延时保存的形态学观察

目的 探索对供肝具有较长时间保护作用的器官保存液。方法 20只新西兰大白兔(体重2．5～3．0kg)随机分为实验组(15只)和实验对照组(5只)。用自制的器官保存液(GPC-Ⅱ-3液)，低温冷藏连续保存兔肝0～192h，观察其组织结构的变化。结果 保存0～120h的兔肝，其组织结构基本正常；保存到144h兔肝，电镜下见肝细胞的线粒体出现轻度肿胀和嵴少：随着保存时间的延长肝细胞出现明显的细胞变性。结论 用GPC-Ⅱ-3液以低温冷藏的方法能够在组织结构上保存兔肝120h。     

脑死亡供体肺脏病理改变及临床移植应用前景探讨

目的观察脑死亡供体肺病理改变,探讨临床移植应用的可行性。方法对23例脑死亡供体肺进行了病理活检,HE染色、网状纤维染色及PASM染色观察肺脏组织变化,电镜观察超微结构变化。结果气管、支气管及肺泡结构尚完整,仅见局灶肺泡上皮细胞及支气管纤毛上皮细胞脱落,未见肺透明膜形成;肺泡间隔未见明显增宽、血管充血易见,血管内皮细胞未见明显改变,未见纤维化表现。电镜下肺泡Ⅱ型上皮细胞胞质轻微水肿,但结构尚完整;内质网扩张,基底膜未见明显改变。结论严格限定纳入标准,脑死亡供肺可以实施移植。     

锌离子在小鼠肺组织分布的TSQ荧光研究

目的研究游离锌离子在小鼠肺组织的定位分布。方法应用TSQ荧光技术检测小鼠肺组织内的锌离子。结果荧光显微镜下可见被TSQ荧光标记的锌离子广泛的分布在肺叶支气管壁粘膜、粘膜下及软骨,肺内小支气管基膜,肺泡上皮和肺内血管平滑肌,尤其以肺叶支气管粘膜以及软骨中荧光染色最为明显。结论小鼠肺组织内含有丰富的锌离子可能参与肺泡上皮细胞以及肺各级支气管软骨细胞中某些蛋白质的合成和组装。     

自制GPC-II-3液对兔肝低温延时保存的形态学观察

目的探索对供肝具有较长时间保护作用的器官保存液。方法20只新西兰大白兔(体重2.5～3.0kg)随机分为实验组(15只)和实验对照组(5只)。用自制的器官保存液(GPC鄄II鄄3液),低温冷藏连续保存兔肝0～192h,观察其组织结构的变化。结果保存0～120h的兔肝,其组织结构基本正常;保存到144h兔肝,电镜下见肝细胞的线粒体出现轻度肿胀和嵴少;随着保存时间的延长肝细胞出现明显的细胞变性。结论用GPC鄄II鄄3液以低温冷藏的方法能够在组织结构上保存兔肝120h。     

缺血再灌注肺脏超微结构变化的电镜观察

目的　观察研究新西兰兔肺缺血 再灌注 (ischemic-reperfusion ,I R)过程肺毛细血管及肺泡上皮细胞超微结构变化。方法　取用新西兰兔I R模型的肺组织 (正常组、缺血 1h、再灌注 0 5h和再灌注 2h ,共 4组 )制作常规电镜标本 ,在透射电镜下进行观察。结果　缺血 1h肺毛细血管和肺泡上皮开始发生病理学变化 ,缺血后再灌注 0 5h时 ,结构明显损害 ,再灌注 2h后 ,毛细血管与肺泡结构均开始表现出损伤的修复趋势。结论　肺I R状态下 ,毛细血管内皮细胞和肺泡上皮细胞形态结构上的异常是导致肺功能障碍的病理学依据。     

人胎肺发育过程中生存素表达及生物学意义

目的检测生存素(survivin)和caspase-3在人胎肺发育过程的表达特征,探讨两者在胎肺发育中的意义。方法采用16～35周人胎肺组织,用免疫组化SP法检测survivin和caspase-3的表达。结果在胎肺的假腺期和小管期,survivin主要表达于远端支气管上皮,在原始肺泡期,survivin阳性表达于原始肺泡上皮,到35周,近端支气管上皮细胞中出现散在的survivin阳性细胞。caspase-3在支气管上皮的表达,以小管末期最强,主要定位于近端支气管上皮、Ⅱ型肺泡细胞。结论 胎肺发育早期,survivin对支气管树形态构筑过程中上皮细胞的发育与分化具有重要的保护意义,caspase-3参与支气管树的重建。胎肺发育晚期,Survivin和caspase-3协同作用,调控着Ⅰ型和Ⅱ型肺泡细胞的成熟和分化。     

小鼠肺泡发育与肺泡上皮细胞分化的电镜观察

目的　观察小鼠胚胎及生后肺泡发育及肺泡上皮分化。方法　小鼠胚胎 14天至生后 14天肺组织 ,隔天取材 ,HE染色光镜观察及透射电镜观察。结果　胚胎 14～ 18天 ,小鼠肺的发育以支气管树分支和管壁结构的逐渐完善为主 ,终蕾上皮为柱状或立方状的未分化细胞。胚胎 19天 ,支气管远端形成许多内壁光滑的原始肺泡 ,其上皮分化出Ⅱ型肺泡细胞。生后 1～ 4天 ,肺泡上皮出现少量扁平的Ⅰ型肺泡细胞 ,但仍以Ⅱ型肺泡细胞为主。生后 5～ 14天 ,成熟肺泡形成 ,肺泡上皮以Ⅰ型肺泡细胞为主。结论　出生时 ,小鼠的肺发育只完成了其大体形态的发生 ,肺泡上皮以Ⅱ型肺泡细胞为主。成熟肺泡的形成、数量的增加及Ⅰ型肺泡细胞的大量出现持续到出生后。     

棉子糖低钾右旋糖酐液供体肺灌注保存的临床病理学研究

目的探讨棉子糖低钾右旋糖酐液(raffinose-low-potassium dextran solution,R-LPD液)供肺灌注低温保存后的组织形态学变化。方法应用光镜、电镜和免疫组化技术对R-LPD液灌注的18例肺移植供肺标本按不同的时间段进行观察。结果R-LPD供肺灌注低温保存的最佳时间是6~8h,随着保存时间延长,供体肺的形态结构改变也逐渐明显,保存30h时,可见肺泡壁及血管周围间质组织水肿,肺泡间隔断裂,部分肺泡上皮细胞变性、坏死、脱落。结论R-LPD液在肺移植冷缺血保存期中起较好的肺泡上皮细胞保护作用,比低钾右旋糖酐液(low-potassium dextran solution,LPD液)具有明显的优越性。     

α1,3半乳糖转移酶基因敲除巴马克隆猪与核供体巴马猪心、肺解剖学与组织学的比较

目的探讨α1,3半乳糖转移酶(GGTA1)基因敲除巴马克隆猪与核供体巴马猪心、肺解剖学与组织学发育变化的异同。方法选取GGTA1基因敲除巴马克隆猪2头(BM164和BM155)及核供体巴马猪1头(BMM2),利用大体解剖学和常规石蜡切片HE染色技术,对其心脏和肺脏进行解剖学与组织学比较研究。结果核供体成年健康巴马猪心脏呈倒置圆锥形,前缘凸,后缘短而直,心表面冠状沟和左右纵沟清晰可见;心壁结构完整,可分3层,心内膜较薄,心室内膜下有较多浦肯耶纤维,心肌排列紧密,横纹清晰,闰盘少且不清楚,心外膜较厚,无髓神经纤维明显。GGTA1基因敲除巴马克隆猪与核供体巴马猪相比较,心脏在解剖学与组织学构造上均未见明显差异。核供体成年健康巴马猪肺脏呈粉红色海绵状,质软而轻,富有弹性;肺脏小支气管软骨丰富,黏膜下层内气管腺明显,可见炎性细胞,细支气管黏膜皱褶呈指状向管腔凸起,周围有炎性细胞浸润,肺泡大小较一致,肺泡壁较厚,尘细胞少见。GGTA1基因敲除巴马克隆猪肺脏解剖学与组织学结构与核供体巴马猪基本一致。结论 GGTA1基因敲除巴马克隆猪与核供体巴马猪心、肺解剖学构造无明显差异,组织学结构正常,心、肺发育良好。     

哮喘大鼠肺内SP阳性纤维的变化和Fos蛋白表达的形态学研究

为探讨P物质(SP)和Fos蛋白与哮喘发病的关系,本实验采用免疫组织化学方法及计算机图像分析技术对正常对照大鼠和哮喘大鼠肺内SP免疫阳性纤维的分布变化及Fos蛋白的表达情况进行了研究。结果显示:(1)正常对照组大鼠肺组织内可观察到SP阳性纤维分布于支气管至终末细支气管和肺血管壁内,但呼吸性细支气管和肺泡壁内少见SP阳性纤维。哮喘组大鼠肺内SP阳性纤维的数量及分布均发生了明显变化,从肺内支气管到终末细支气管均有较密集的SP阳性纤维呈束走行;且呼吸性细支气管和肺泡壁内也出现了SP阳性纤维。哮喘组大鼠肺内SP阳性纤维的数量明显高于正常对照组大鼠(P<0.05)。(2)正常对照组大鼠的肺内几乎不表达c-fos基因,但在哮喘组大鼠肺内可观察到许多Fos阳性细胞,这些细胞主要为炎性细胞和平滑肌细胞。以上结果表明,肺内SP阳性纤维增生及c-fos基因表达与哮喘发病密切相关。     

Short term morphologic effects of high ambient levels of ozone on lungs of rhesus monkeys.

Groups of laboratory-reared, young adult rhesus monkeys were exposed to 0.8 p.p.m. or 0.5 p.p.m. of ozone for 8 hours a day on 7 consecutive days. Lesions were studied using correlated techniques which permitted examination of specified levels of airways and adjacent lung parenchyma by light microscopy, scanning electron microscopy, and transmission electron microscopy. Lesions were observed in the trachea and lungs of all exposed animals. The extent and severity of damage, but not its nature, varied with exposure concentration. Damage was most severe in respiratory bronchioles and more distal parenchymal regions were unaffected. Major features of the response within respiratory bronchioles were hyperplasia and hypertrophy of nonciliated bronchiolar epithelial cells and intraluminal accumulations of macrophages. Replacement of type 1 epithelium in alveoli by type 2 cells and forms intermediate between types 1 and 2 were also observed. In large conducting airways, damage to ciliated cells was observed but mlcus-producing cells were morphologically unaltered. Two gradients in severity of ozone-induced lesions were appreciable in the trachea and lungs. The most obvious gradient was in respiratory bronchioles where the degree of damage was most severe in proximal locations. A second gradient in severity was noted in conducting airways in which more severe and extensive lesions occurred in the trachea and major bronchi than in small bronchi and terminal bronchioles.     

Stereological estimation of the volume weighted mean volumes of alveoli and acinar pathways in the rat lung to characterise alterations after ischaemia/reperfusion

The aim of this study was to characterise pulmonary reimplantation injury in isolated, perfused rat lungs following 2 h of cold ischaemia, and 50 min. of in vitro reperfusion. The effects of 2 differently composed lung preservation solutions (low potassium Euro-Collins and Celsior; each n=5) were examined in comparison with untreated, nonischaemic control lungs (n=3). After fixation by vascular perfusion and tissue collection by systematic random sampling, the volume weighted mean volume (_v) of alveoli and acinar pathways was estimated by light microscopic stereology using the method of point sampled intercepts in plastic embedded, Azan-stained material. Significantly higher _v of alveoli and acinar paths was found in the Celsior group than in Euro-Collins preserved lungs. However, in the controls the size of acinar pathways was similar to Celsior preserved lungs whereas alveolar size was comparable to preservation with Euro-Collins. The between-animal coefficient of variation of alveoli was very low in controls and Celsior preserved but higher in the Euro-Collins group. Size distribution of alveoli and acinar paths in 15 size classes was largely homogeneous in all groups tested. In the Euro-Collins group the fractions of both class 1-alveoli and class 1-acinar paths significantly exceeded those of the other groups. Widely expanded alveoli (size classes 13–15) only occurred after preservation with Celsior whereas wider acinar paths (size class 15) were found in the Celsior group and in the controls. It is concluded that lung preservation with low-potassium Euro-Collins and Celsior solutions may act differently on distinct spaces in the distal gas-exchange regions of lungs. This may be due to selective effects on pulmonary surfactant activity and on elastic tissue elements in the alveolar ducts, respectively. Additionally, the method of point sampled intercepts is considered to be an efficient tool to evaluate the effects of different preservation solutions on lung parenchyma.     

The effect of changes in lung volume on the size and shape of alveoli

1. A technique is described by which living anaesthetized guinea-pigs were artificially ventilated with a positive and negative pressure cycle, and rapidly frozen at the instant of cessation of ventilation at a preselected point on the respiratory cycle.2. The dimensions of alveoli and alveolar ducts were measured by histological morphometric methods, and related to the degree of lung inflation at the instant of freezing.3. The alveolar volume fraction increased from 0.52 at low lung volumes, up to 0.62 at lung volumes in the mid-inflation range, then decreased to 0.51 at high lung volumes. The alveolar duct volumetric fraction remained constant at all lung volumes.4. The total volume of alveoli increased linearly with increase in lung volume. The total alveolar duct volume increased little until the lungs were 40% inflated, above which it increased steeply.5. The total number of alveoli was linearly related to the body weight.6. The total alveolar surface area increased steeply between lungs of low lung volume and those which were up to 50% inflated, above which the increase levelled off. The absolute values of total alveolar surface depended on the body weight as well as the degree of lung inflation.7. The mean alveolar duct diameter was 40% greater, and the mean alveolar mouth diameter 35% greater in lungs which were fully inflated than in lungs which were nearly collapsed. The average geometrical shape of alveoli was not related to the degree of lung inflation.8. The harmonic mean thickness of the air-blood barrier was 33% less in lungs which were fully inflated than in lungs which were nearly collapsed. The alveolar surface membrane was smooth whether the lungs were fully inflated or collapsed.     

Insulin-like growth factors in lung precancer

Investigation of IGF-system resulted in distinguishing two groups of processes depending on the size of the bronchi. Goblet cell metaplasia, basal cell hyperplasia with or without atypical features, squamous cell metaplasia, dysplasia 1-3, neuroendocrine cell hyperplasia, epithelial atrophy were found in the large bronchi. The same changes occurred in the bronchioli and alveoli plus adenomatous hyperplasia with or without atypia, oval and narrow structures with epithelial atrophy, atypia and proliferation. Common property was accumulation in the cells of various members of IGF system: high accumulation of IGF II, IGFBP 1,2,3,4 in dysplasia; the same plus IGFBP 11,2,3,4,5 in squamous metaplasia. Tuberculous granuloma cells are an important source of IGF II, IGFBP 2,3,4,5,6 playing a role in proliferation and apoptosis of the pulmonary epithelium.     

Motility of human polymorphonuclear leukocytes. Roles of hydroxy fatty acids, other lipids, and cations.

Neonatal lung morphology was evaluated in 12 rhesus monkeys delivered by caesarean section 1 month before term and cared for as human premature neonates. In 6 monkeys, 0.20 to 0.27 ml of natural rabbit surfactant (SA) was instilled intratracheally before the first breath; the other 6 served as controls. Histology and morphometry of the controls' lungs revealed changes typical of hyaline membrane disease (HMD) in human premature infants, whereas the SA-treated lungs showed improved alveolar expansion and only minor lesions typical of HMD. Transmission and scanning electron microscopy of the controls' lungs showed extensive necrosis and desquamation of bronchiolar epithelium, with formation of hyaline membranes; Type I alveolar epithelial cells showed lesions similar to those in bronchioles, but immature Type II cells appeared relatively well preserved. In the lungs of SA-treated animals, the epithelial lining of most airways and alveoli was intact. This first demonstration of the beneficial effect of exogenous SA on lung adaptation in premature primates indicates that prophylaxis with SA might prevent HMD in premature human infants.     

Morphology of aging lung in F344/N rat: alveolar size,connective tissue,and smooth muscle cell markers

This study investigated the morphological changes of lungs in F344/N rats (9-36 months old). We initially examined general and quantitative morphological changes, and then we used immunohistochemistry to detect distributional changes in collagen subtypes (types I, III, and IV) and smooth muscle cell (SMC) markers (alpha-smooth muscle actin (ASMA), gamma-smooth muscle actin (GSMA), desmin, and vimentin) in the lungs. In 24-month-old rats, alveolar ducts and alveolar sacs were enlarged, and alveoli were wider and shallower than in younger animals. In old rats (>/=27 months), terminal and respiratory bronchioles and alveolar ducts were dilated and alveoli were more extended than in 24-month-old rats. No age-related distributional changes were observed for collagen types I, III, and IV as revealed by immunohistochemistry, or elastin as revealed by resorsin fuchsin. SMCs in the extra- and intrapulmonary bronchi were immunoreactive for ASMA, GSMA, and desmin, but not for vimentin at all ages. In old rats (>/=27 months), SMCs were loosely arranged in comparison with younger animals, and stainability for GSMA and desmin was decreased. In the respiratory bronchioles and alveolar ducts, a few cells immunoreactive for ASMA and vimentin were observed in the smooth muscle aggregations of the alveolar orifice in rats younger than 12 months. In older rats (>20 months), cells immunoreactive for ASMA and vimentin were increased in septal tips. In conclusion, extension of distal airways and immunohistochemical changes of SMC markers in F344/N rat lungs were evident by approximately 24 months of age, but there was no apparent change in connective tissue morphology.