低能量He-Ne激光照射体外循环血液对IDDM患者红细胞膜ATPase活性的影响

本文研究了低能量He-Ne激光照射体外循环血液对IDDM患者红细胞膜ATPase(Na~+/K~+-ATPase,Ca~(2+)-ATPase、Mg~(2+)-ATPase)活性的影响,结果表明,IDDM患者膜泵功能活性显著降低;低能量He-Ne激光照射血液可激活膜ATPase活性。作者讨论了膜泵功能降低及激光作用的可能机制。     

蝌蚪皮肤细胞中磷酸酶定位的电子显微镜研究

本实验以花背蟾蜍后肢芽期的蝌蚪为材料,研究酸性磷酸酶及碱性磷酸酶在表皮细胞中定位。酸性磷酸酶在细胞表面个别粘液泡、细胞间连接复合体及细胞间隙上都有分布。在细胞内此酶主要定位于溶酶体、高尔基复合体及内质网上。碱性磷酸酶主要定位于细胞表面的微绒毛、细胞间隙、连接复合体及胶原纤维上。     

免疫胶体金标记定位昆诺藜中的胼胝质和胼胝质降解酶

运用免疫胶体金技术研究了感染蚕豆萎蔫病毒2号以及健康昆诺藜细胞中胼胝质和胼胝质降解酶的分布情况。电镜观察发现胶体金特异性标记在含有VP37小管的胞间运动通道周围,其余区域感病和健康细胞的胼胝质及胼胝质降解酶分布差别不大,胼胝质主要分布于细胞间隙的大块电子致密物质以及细胞壁结构发生较大变化的区域,胼胝质降解酶主要分布于筛板、维管束细胞紧贴细胞壁区域的大块电子致密物质上,在细胞壁结构发生较大改变的区域也有胼胝质降解酶的分布。推测胼胝质在昆诺藜细胞中的积累和降解可能与细胞壁结构成分改变有关。     

失血性休克下犬重要器官超微结构的观察

犬失血性休克五小时,在电镜下可见心、肺、肝、小肠和肾等重要器官出现明显的结构损伤,具体变化如下:心肌纤维水肿,核周胞浆增多,肌原纤维粗细不均,间距变大,肌丝分散并有溶解;闰盘变直,有不同程度的肌丝解离;肌质网和横小管扩张;线粒体凝集变性,基质局灶性空泡化,外膜破裂,偶见巨大线粒体(图1)。肺泡内含水肿液;肺泡Ⅰ型细胞肿大,线粒体肿胀,肺泡Ⅱ型细胞增多,嗜锇性板层小体减少或变空,肺泡隔内毛细血管内皮细胞肿胀,基膜变薄,腔内有血小板、粒细胞聚集及微血栓形成(图2)。肝细胞索断裂,肝细胞游离在极度扩张的Disse间隙和肝窦内,肝细胞变大变圆,表面微绒毛消失,细胞膜破裂,线粒体高度肿胀,嵴断裂或消失,可见巨大线粒体,内含副结晶,不少肝细胞核浓缩,     

结肠癌相关抗原LEA分布的免疫电镜和免疫光镜研究

本文应用免疫细胞化学技术对新研究成功的结肠癌相关抗原LEA在光镜、电镜下进行了较详细的定位观察研究。LEA是一种可溶性抗原,其表达与癌细胞的分化程度密切相关。应用ABC免疫酶标法发现LEA在癌组织内的分布呈明显的异质性,LEA存在于癌细胞腔缘的质膜上,有明确的极性。在培养细胞,LEA均匀分布在膜表面。免疫电镜观察到LEA多集中分布在微绒毛上,金颗粒标记在细胞外被的糖萼成分上。LEA可能是通过微绒毛结构排出到腺腔中去的。     

对杯状细胞分泌方式的扫描电镜观察

本文应用扫描电镜对家兔气管、支气管、十二指肠、回肠与胆囊上皮内杯状细胞的分泌方式和过程进行了观察。发现在休止期杯状细胞表面低于其相邻的上皮细胞,顶部有长而密集的微绒毛。以后随分泌颗粒的形成和增多,逐渐向细胞顶端转移、集聚并压挤细胞顶部的细胞膜,使之逐渐向外膨出,微绒毛亦相继变短、变粗、数目减少终于消失。在细胞表面的细胞膜上出现小孔。然后胞膜局部下陷,小孔增多,状如筛孔,最后细胞表面变平。根据上述现象推断杯状细胞是以胞吐方式进行分泌,其表面微绒毛起到膜储备的作用。     

棉铃虫 H.Armigera 和大蓑蛾 C.Variegata 中肠上皮细胞微绒毛形态结构研究

用超薄切片及冷冻蚀该制样方法,对大蓑蛾C.Variegata中肠和棉铃虫H.Armigera中肠的超微形态结构研究中,发现两者的中肠微绒毛内含物和已报导的豚鼠,家兔胃内壁上皮细胞微绒毛内含有明显差异,现简述如下:大蓑蛾C.Varigata中肠冷冻蚀该观察能清楚地看到上皮层杯状细胞和柱状细胞的核、核孔、核膜、细胞膜及微绒毛。从核孔园柱状突出或边缘隆起四周有环形浅窝,核膜面微粒多少,可确定核膜的PF面和EF面以及细绒毛的PF面和EF面。在微绒毛的横断面,可看到排列整齐的线粒体(图略)。另外,棉铃虫H.Armigera中肠超薄切片观察,在杯状细胞和柱状细胞的纵切面和横切面同样看到大量排列整齐的     

5’—核苷酸酶和硷性磷酸酶在二乙基亚硝胺诱发大鼠肝癌细胞中的定位

我们用光镜和电镜观察了5′—核苷酸酶和硷性磷酸酶在二乙基亚硝胺诱发SD大鼠肝癌细胞中的定位,结果发现这两种酶是定位在正常鼠肝细胞毛细胆管微绒毛区和SD大鼠肝癌癌细胞毛细胆管微绒毛区,而且后者(图3、4、5)比前者(图1、2)丰富得多。提示肝癌患者血清中上述两种酶显著增高,可能系因癌变而使酶经毛细胆管附近的相邻两个细胞缝隙连接而进入血液循环。     

胃癌印戒细胞超微结构酶细胞化学研究：五种细胞器标志酶的电镜观察

胃印戒细胞癌多见于30岁以下的女性,其侵袭力强,转移率高,这种生物学行为的发生机理日益受到关注。但从酶学特别是对多种细胞器标志酶进行超微结构水平的研究,文献中未见报告。本文应用酶细胞化学技术对胃癌印戒细胞五种细胞器标志酶(碱性磷酸酶AKPaee,酸性磷酸酶Acpace,葡萄糖-6-磷酸酶G6Pase,焦磷酸硫胺素酶TPPaee和细胞色素氧化酶Coaee)进行了细胞和超微结构水平的定位观察,并以正常胃粘膜作为对照,探讨了胃癌印戒细胞五种细胞器标志酶的分布特点及其与生物学行为间的关系。结果发现:正常胃粘膜上皮细胞AKPace均阴性;Acpase活性以主细胞为最强,旦分布于自噬及分泌溶酶体中;胃粘膜上皮细胞的核周间隙和粗面内质网(RER)G6paee均阳性,主细胞富于RER     

虹膜色素上皮细胞Na^＋—K^＋—ATP酶的超微结构定位

Na+-K+-ATP酶存在于细胞膜,具有进行钾钠离子交换的机能,并与细胞内离子、水物质等的输送有关,因此被称为钠钾泵[1].本文采用电镜酶组化方法对兔虹膜色素上皮细胞Na+-K+-ATP酶的分布进行了研究.     

供肝热和冷保存性缺血性损伤的超微结构观察

目的:探索供肝热、冷缺血对肝组织的超微结构影响,并讨论与某些手术并发症发生的关系.方法:从我院实施的67例原位肝移植病例中选择9例移植前供肝活检组织病理学检查无明显形态学改变、供肝者临床肝功能正常、肝炎病毒及巨细胞病毒检测均为阴性的供肝组织进行透射电镜观察.结果:在供肝中央静脉周围肝细胞质中常有较多吞噬溶酶体,少数肝细胞核出现浅的切迹,胞质中线粒体明显减少,部分线粒体肿胀.少部分毛细胆管腔扩大,微绒毛肿胀,微绒毛芯中微丝束减少或消失.部分小叶间胆管上皮细胞凋亡,管腔中含凋亡小体及细胞碎片.少数小叶间静脉内皮细胞肿胀,胞质空泡变性,甚者内皮细胞脱离管腔,细胞质膜断裂.结论:供肝毛细胆管和小叶间胆管上皮细胞的损伤是保存性损伤引起轻度胆汁淤积的主要因素.而小叶间胆管周围小血管内皮细胞的损伤可能是导致小叶间胆管上皮细胞凋亡的直接原因.小叶间静脉内皮细胞的损伤使血管壁平滑度受损,血流减缓,加重肝细胞的缺氧.     

Cytochemistry of gastric parietal cells with high-pressure freezing followed by freeze-substitution

Gastric parietal cells were examined for changes in their ultrastructure and distribution of the proton pump during feeding and fasting states in rats. The fundic glands from rats fed ad libitum or fasted with free access to water were cryofixed using high-pressure freezing followed by freeze-substitution in acetone containing osmium or acrolein and then embedded in Epon 812 or Lowicryl K4M resin, respectively. Excellent ultrastructural preservation was achieved. During the feeding state, intracellular canaliculi and numerous microvilli were well developed, while tubulovesicles were poorly developed. In contrast, during the fasting state, the microvilli in the narrowed space of the intracellular canaliculi were tightly packed and the tubulovesicles were enlarged. Ultrathin sections were immunostained with antibodies against the alpha- and beta-subunits of the proton pump, H+ x K(+)-ATPase, using the immunogold method. The labelling was strong and clearly localized in comparison with that obtained using the conventional chemical-fixation method. Each subunit was localized on the membrane of the microvilli, intracellular canaliculi and tubulovesicles. The distribution of subunit proteins varied between the two states. During ad libitum feeding, the immunolabelling was localized strongly on the membranes of the microvilli and intracellular canaliculi. In contrast, the labelling was strong on the tubulovesicle membrane in the fasting state. The results obtained with each anti-subunit antibody by H+ x K(+)-ATPase immunostaining revealed differences in distribution and labelling density between the feeding and fasting states.     

Morphological and histochemical analyses of living mouse livers by new 'cryobiopsy' technique

A new 'cryobiopsy' (CB) technique has been invented for freezing the functioning livers of living mice in vivo without stopping their blood circulation. Livers of anesthetized mice were pinched off with pre-cooled CB forceps and immediately plunged into isopentane-propane cryogen. They were routinely freeze-substituted in acetone containing paraformaldehyde for light microscopy (LM) or osmium tetroxide for scanning electron microscopy (SEM). By freeze-fracturing some of them with a scalpel in liquid nitrogen before the freeze-substitution, well-preserved tissue areas were exposed only for SEM. They were either embedded in paraffin wax for LM or infiltrated with t-butyl alcohol followed by freeze-drying for SEM. Serial paraffin sections were stained with hematoxylin-eosin (HE) or histochemical periodic acid-Schiff (PAS) reaction. By HE-staining, the tissue surface areas were often compressed with the CB forceps and sinusoidal erythrocytes became aggregated side by side. In slightly deeper tissue areas, however, hepatic sinusoids were widely open with flowing erythrocytes. Lots of PAS-reaction products were well preserved in hepatocytes of the CB specimens. On the contrary, they were unevenly distributed in hepatocytes of conventionally quick-frozen specimens, and often lost in those of the conventionally dehydrated specimens. By SEM, some cell organelles, such as mitochondria and endoplasmic reticulum, and also dilated fenestrae of endothelial cells, open Disse's spaces and bile canaliculi appeared to be under normal blood circulation in the prepared CB samples. The new CB technique would be easy and useful for repeated examination of functioning organs of a living animal.     

毛细胆管发育不良致先天性阻塞性黄疸的超微结构观察

用电子显微镜观察了一例先天性阻塞性黄疸病例的肝脏超微结构,结果显示肝细胞内有不同程度的淤胆,大部分肝细胞间未见毛细胆管结构或毛细胆管发育不良,提示本例患者的阻塞性黄疸由先天性毛细胆管发育不良引起,电镜检查有利于判断患儿的预后。     

Acetylcholine-induced activation of the eccrine sweat glands in a case of hypohidrotic congenital ectodermal dysplasia

In a male patient suffering from congenital ectodermal dysplasia with sweating only on the plantar and palmar surface, we found that the anhidrotic eccrine sweat glands became able to produce sweat after repeated local application of acetylcholine. We ultrastructurally examined the sweat glands in skin biopsies before and after acetylcholine treatment. The secretory portion of the sweat glands of the right plantar region was characterized by well-developed basal infoldings and intercellular interdigitations, and also by dilated intercellular spaces and canaliculi. In the flexor region of the right brachium (anhidrotic area), the secretory portion showed hypoplastic features, especially denoted by the presence of few locally distributed basal infoldings and intercellular canaliculi. The secretory cells seemed to be hypofunctional with the nuclei containing much heterochromatin. After treatment, in the flexor region of the left brachium and the right infraclavicular region, where sweating was induced, the secretory portion appeared to be considerably activated, though hypoplastic structures were unchanged: the secretory cell nuclei contained dispersed chromatin, the Golgi apparatus in the dark cells was well developed, and both in the clear cells and the myoepithelial cells micropinocytotic vesicle-like structures increased in number. In ductular cells, mitochondria also increased in number.     

Ultrastructural transformation of gastric parietal cells reverting from the active to the resting state of acid secretion revealed in isolated rat gastric mucosa model processed by high-pressure freezing

To elucidate a functional transformation of gastric parietal cells, we have newly developed an isolated rat gastric mucosa model whose parietal cells exhibited a reverting process from the active to the resting state of acid secretion. Briefly, the parietal cells were treated with cimetidine following prior stimulation of acid secretion in the model, and cryofixed by plunge freezing for light microscopy or high-pressure freezing for electron microscopy. As a result, immunohistochemistry of H(+)/K(+)-ATPase demonstrated a progressive translocation of H(+)/K(+)-ATPase from the apical to the cytoplasmic region. The ultrastructure of parietal cells at 5 min in the reverting phase was quite similar to that of maximally stimulated one. However, the apical microvilli of intracellular canaliculi (IC) changed bulbous by degrees, resulted in complete occlusion of IC at 60 min in the reverting phase. The apical membranes were subsequently internalized into the cytoplasm forming unique penta-laminar membranes. Interestingly, at 90 min in the reverting phase, the penta-laminar membranes formed a number of multilamellar autophagosomes that were intensely labeled for H(+)/K(+)-ATPase. Then, the parietal cells exhibited well-developed Golgi apparatus and lysosomal compartments involving the multilamellar membranes at 105 min, and mostly reverted to their resting conformation at 120 min in the reverting phase. Corresponding to the ultrastructural changes of microvilli, the immunohistochemistry of ezrin showed a dissociation of ezrin from the apical region at 30 min in the reverting phase. The present findings provide new insights into the functional transformation in gastric parietal cells reverting to their resting conformation.     

大鼠侧脑室及其脉络丛室管膜上皮的超微结构观察

采用扫描和透射电子显微镜研究大鼠侧脑室及其脉络丛室管膜上皮的超微结构特征，并探讨其功能。扫描电镜观察结果：侧脑室壁室管膜上皮细胞游离端呈不规则多边形，可见纤毛及微绒毛，偶见分泌泡，脉络丛上皮细胞游离端微绒毛及分泌泡丰富，纤毛光见，可见、丛上细胞”。透射电镜观察发现，侧脑室壁及脉络丛上皮均由单层立方上皮组成，但二类上皮细胞的超微结构明显有别，室管膜上皮表面可见纤毛及微绒毛，分泌泡少见，脉络丛上皮游离端微绒毛丰富，常吻合成迷路样网，微绒毛的顶端膨大，形成微顶浆分泌小泡，胞浆内细胞器丰富，大鼠侧脑室壁及脉络丛上皮超微结构的差异反映二者功能的不同，本研究认为侧脑室脑脊液分泌的主要部位是脉络丛，其分泌的方式除脉络丛上皮细胞的主动运输功能外，可能微顶浆分泌法是侧脑室脑脊液形成的主要方式。     

Dubin Johnson综合征的肝细胞超微结构观察

采用肝穿刺的方法,利用透射电镜对Dubin Johnson综合征(结合型高胆红素血症)患者进行了肝细胞超微结构的电镜观察。观察结果：电镜下可见到肝细胞内及毛细胆管周围有大量的外有单层膜包绕的色素颗粒,颗粒大小不均匀;其内容物有三种形态,即电子密度较高且均匀的颗粒,电子密度略低的颗粒及电子密度不均匀的内含有小脂滴样物质的颗粒;毛细胆管扩张,腔内微绒毛减少,管壁增厚;粗面内质网及滑面内质网明显扩张成许多小囊泡,使得肝细胞成筛孔状,肝细胞间有胶原纤维增生,肝细胞间隙增宽,并有微绒毛出现。其结果有助于临床的诊断。     

肠易激综合征患者结肠粘膜细胞超微结构的研究

用电子显微镜观察6例肠易激综合症（IBS）患者,2例慢性结肠炎和2例正常对象结肠粘膜超微结构.结果显示,IBS患者结肠粘膜上皮细胞膜完整,细胞形态无异常,细胞间隙紧密,细胞微绒毛分布不规整,密度低而不均,长短方向不一,但线粒体、粗面内质网等细胞器无规律性和明显的变化.而IBS患者粘膜上皮中杯状细胞、浆细胞、肥大细胞和神经内分泌细胞内均可查见大量的分泌颗粒、空泡和粗面内质网等,呈现功能活跃和分泌旺盛状态.提示：IBS结肠粘膜内肥大细胞、浆细胞、神经内分泌细胞和杯状细胞的超微结构改变可能与IBS发病机制相关.     

The capillary of the organum vasculosum laminae terminalis (OVLT) in rabbits is more permeable to horseradish peroxidase (HRP) than that in rats

The distribution of the tracer, horseradish peroxidase (HRP), in the organum vasculosum laminae terminalis (OVLT) of rabbits and rats was examined by electron microscopy. In rabbits, the HRP-diaminobenzidine reaction products were heavily distributed in the OVLT and surrounding brain tissues 10 and 60 min after the injection of HRP (50 mg kg(-1), i.v.), and were retained in the parenchymal tissues at 24 h post-injection. The majority was found in numerous large phagosomes of macrophages located in the perivascular spaces of the vascular beds and in ependymal cells (tanycytes) in the parenchyma. A large amount of reaction product was also localized in the intercellular clefts of the parenchyma. HRP-incorporation was seen in both nerve cells and ependymal cells in the OVLT at 10 min post-injection, but only in nerve cells in the preoptic area at 60 min post-injection. In rats, however, a small amount of the reaction products was observed in the OVLT 10 min after the injection of HRP (50 and 70 mg kg(-1), i.v.), and the levels were markedly reduced at 60 min post-injection. No HRP-incorporation by nerve cells was seen. From these findings, we concluded that the capillary of the OVLT of the rabbit is more permeable to HRP than that of the rat.