皮肤共聚焦激光扫描显微镜在儿童色素减退性疾病诊断中的应用

目的 探讨儿童常见色素减退性疾病皮肤共聚焦激光扫描显微镜(CLSM)图像的基本特征.方法 分别观察1 915例色素减退性疾病(包括白癜风、特发性滴状色素减少症、无色素痣、线状苔藓、白色糠疹及炎症后色素减退)患者的皮损,用CLSM观察皮损处、交界处及白斑周边正常皮肤的各层镜下特征.应用卡方检验和Fisher精确检验进行统计学分析.结果 1级和2级色素减少占总病例数的51.4％(984/1 915)和35.7％(684/1 915),白癜风3级色素减少的比例为77.9％(141/181),明显高于其他疾病(0～ 10.3％);815例角质层轻度角化过度,1 060例棘层灶性水肿,79例基底细胞环出现改变,1 133例真皮浅层可见稀疏炎症细胞或树枝状细胞.6种色素减退性皮肤病CLSM扫描图像色素减少程度及各层CLSM图像变化均有统计学差异(P＜0.05).CLSM特征:特发性滴状色素减少症可见色素环完全缺失;线状苔藓、白色糠疹、炎症后色素减退可见非特异性炎症改变,线状苔藓可见点灶状基底细胞液化变性;无色素痣仅为色素减少及折光变弱,白癜风白斑区色素完全缺失,且皮肤色素环缺乏完整性.结论 白癜风、特发性滴状色素减少症、无色素痣、线状苔藓、白色糠疹及炎症后色素减退皮损CLSM图像有差异,可以作为鉴别诊断的依据之一.     

子宫内膜增生及子宫内膜癌的内分泌特征、甾体激素受体及超微结构研究

对４８例子宫内膜增生，２４例子宫内膜癌进行了血甾体激素放免分析及组织雌、孕激素受体生化定量检测，对３８例子宫内膜增生及２２例子宫内膜癌进行了受体免疫组化定性检测，对６例子宫内膜增生及１２例子宫内膜癌进行了扫描电镜及透射电镜观察。结果表明，子宫内膜增生与内膜癌之间血甾体激素水平无明显差别。但绝经后的子宫内膜增生及内膜癌患者的雌激素水平高于正常绝经后的妇女；从子宫内膜增生，腺囊样增生，腺瘤样增生到内膜癌，其雌、孕激素受体含量逐渐降低；受体含量与临床分期、细胞分化程度、宫颈浸润、肌层浸润及复发情况相关。经对雌、孕激素受体的定性分析，子宫内膜增生阳性率近１００％，非典型增生及原位癌为８３．３３％，内膜癌为７５％及８１．２５％。电镜观察子宫内膜增生的雌激素依赖性结构明显增加，而内膜癌的雌激素依赖性结构减少，细胞器形态排列异常伴有核异型性及核小体，并出现假包涵体。     

豚鼠耳蜗外毛细胞的共聚焦图像

建立了用激光扫描共聚焦显微镜观察豚鼠耳蜗外毛细胞生理变化及细胞形态的方法。以荧光染料Ｆｌｕｏ－３作胞内Ｃａ￣（２＋）指示剂，用共聚焦显微镜观察了乙酰胆碱（ＡＣｈ）、ＡＴＰ、高钾引起的外毛细胞（ＯＨＣ）胞内Ｃａ￣（２＋）浓度的变化。三种物质均引起Ｃａ￣（２＋）升高，但升高的幅度、时相及所在细胞部位不同。高钾还引起ＯＨＣ短缩、偏折。利用共聚焦显微镜“光学切片”的特性，对Ｆｌｕｏｒｅｓｃｅｉｎ荧光素染色的活的ＯＨＣ形态及碘化吡啶（ＰＩ）染色的细胞核形态进行了三维重建。讨论了共聚焦显微镜在ＯＨＣ生理及形态学研究中的应用前景。     

高级别子宫颈上皮内瘤变宫颈锥切及子宫全切除标本病变组织的病理特点

目的 探讨高级别宫颈上皮内瘤变(CIN,即CIN Ⅲ、Ⅲ)官颈锥切及子宫全切除术病变组织的形态特征、分布、生长方式及衍化特点等病理特点,为宫颈锥切治疗的深度和范围提供依据.方法 选择高级别CIN宫颈锥切标本及子宫全切除标本共592份,肉眼观察宫颈外口病变的形态特征,镜下观察高级CIN病变在各象限的分布特点.随机抽取其中200份标本,镜下观察其病变的深度、宽度及程度;观察病变细胞的特征、生长方式、衍化特点以及病变周围的间质反应情况.结果 (1)肉眼观察:宫颈光滑46份(7.8%),轻度糜烂151份(25.5%),中度以上糜烂395份(66.7%);合并宫颈肥大48份(8.1%)、宫颈赘生物4份(0.7%).(2)镜下观察:高级CIN病变在宫颈各象限的分布比例比较,差异无统计学意义(P>0.05).累及腺体深度<3 mm者188份(94.0%),3～5 mm者12份(6.0%),无一例>5 mm;累及腺体的宽度最宽为1.9 mm;累及腺体程度:孤立性累及腺体者77份(38.5%)、多灶性累及腺体者143份(71.5%)、融合性累及腺体者73份(36.5%);生长方式:平铺性生长162份(81.0%),内陷性生长98份(49.O%),外生性生长13份(6.5%);衍化特点:移行变化66份(33.O%),骤然变化167份(83.5%).高级别CIN有多种细胞类型,多数为基底细胞型,小圆细胞型;并有28.5%-45.0%的标本其病变周围有不同类型的免疫间质反应.结论 肉眼观察,高级别CIN病变无具有诊断意义的特征性改变;镜下观察,高级别CIN病变以平铺性生长、多灶性累及腺体以及累及腺体深度<3 mm为主,提示高级别CIN病变行宫颈锥切时既要注意切除的深度,又要注意切除面的范围.     

电子显微镜与聚合酶链反应技术检测阴道及子宫颈人乳头瘤病毒

应用电子显微镜（电镜）、聚合酶链反应（ＰＣＲ）技术，对１９９０年４月至７月，因外阴瘙痒、白带多就诊检查发现阴道下１／３或子宫颈有异常突起，病理及核酸分子杂交技术检查证实为人乳头瘤病毒（ＮＰＶ）感染的标本４７份，进行ＨＰＶ检测。ＰＣＲ检测阴道尖锐湿疣ＨＰＶ６，１１的阳性率为９２．６％，ＨＰＶ６，１１和ＨＰＶ１６，１８混合感染阳性率为３．７％。电镜下未观察到典型的ＨＰＶ颗粒，观察到挖空细胞核内存在着染色质之间颗粒及染色质周围颗粒，在３例斑点杂交ＨＰＶ１１阳性者的标本中观察到核内小体。基底细胞也有改变。     

Solid-state transformation of a leukotriene antagonist

SK&F 104353, a potent leukotriene antagonist, exists in two physically distinct species as a disodium salt. Type I melts with decomposition, showing a single endotherm in the region of 250–270°C in differential scanning calorimetry (DSC) thermograms, as well as exhibiting a weak powder X-ray diffraction (XRD) pattern. Type II contains a single DSC melting endotherm in the same region, but in contrast has additional, smaller endotherms in the region of 70–85°C; the new transitions remain upon cooling and reheating. The powder XRD pattern of II is very similar to that of I with respect to peak positions, but II's bands are sharper, suggestive of increased crystallinity. Types I and II can also be distinguished using solution calorimetry, manifesting different heats of solution in an ethanol/water mixture. Apparent irreversible conversion of I to II can readily occur, and has a significant impact with respect to physical stability of potential suspension aerosol formulations. The results are explained in terms of a solid-state transformation, i.e., from a metastable, partially amorphous form of low crystallinity (I) to a similar but more physically stable form of higher crystallinity (II); the conversion is facilitated by the presence of water, although true ‘hydration’ is not involved. Temperature-dependent Raman spectroscopy has been applied to characterize the transformation in more detail. The unusual solid-state behavior of this compound may reflect more general structural characteristics of leukotriene antagonists.     

An electron microscopic study of Aujeszky's disease

Summary Sixteen calves were killed at intervals during the course of the disease from 48 h onwards after subcutaneous infection with Aujeszky's disease virus. Ultrastructural changes were evident in the spinal ganglia from 84 h post-inoculation and the intercostal nerves from 96 h post-inoculation. The cytopathic changes in the spinal ganglia consisted of neuronal degeneration, neuronophagia, Schwann cell degeneration and cellular infiltration. The neuronophagic nodule was invariably contained within an intact sheath of satellite cells. Changes in the intercostal nerves were less dramatic but cellular infiltration was frequently seen and occasional Schwann cells were degenerate. In the terminal stages of the disease demyelination was rarely observed. In the ganglion virus was invariably seen in degenerating neurons and occasionally in Schwann cells and monocytes. Satellite cells were rarely infected even when ensheathing an infected neuron. Extra-cellular virus was not observed in ganglia or nerves. Schwann cells and monocytes in the nerves were occasionally infected. Virus particles were seen in the axoplasm both in the ganglion and in the entire length of the nerve. The particles in the axoplasm varied in morphology; thus unenveloped and enveloped particles, and particles in the process of acquiring an envelope were recognised. It was concluded that the neural pathway of Aujeszky's disease virus is probablyvia the axoplasm.     

Molecular and ultrastructural studies of non-crystalline calcium phosphates

Five non-crystalline calcium phosphate precipitates prepared from solutions varying in pH, degree of supersaturation and carbonate content were examined by physicochemical and morphological methods. X-ray diffraction, infrared spectrophotometry and electron diffraction measurements confirmed the amorphous nature of each precipitate. Both transmission and scanning electron microscopy demonstrated that the native elemental particles of all 5 synthetic products were spherical in shape and independent of the preparative procedures employed. Four of the precipitates contained spherules of similar sizes (200–1200 Å in diameter) while the one relatively rich in carbonate exhibited much larger particles (700–2000 Å in diameter). The spherules were uniformly dense to the electron beam when viewed with the current density at its lowest useful level. Exposure to higher electron currents immediately resulted in the development of numerous electron-lucent centers in each particle. Heating in vacuum to temperatures up to 600° did not change the amorphous nature of the precipitates or the morphology and size of their elemental particles even though almost all of the water contained in the original precipitates was removed by this process. Heat treatment did not induce formation of electron-lucent centers. On the contrary it, made the spherules increasingly resistant to beam damage. These findings suggest that the beam electrons interact with the oxygen or hydrogen atoms of the aqueous constituent of the amorphous calcium phosphate and that this initial reaction is followed by displacement and/or loss of other atoms.

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Zusammenfassung Fünf nicht-kristalline Calciumphosphat-Niederschläge, welche aus Lösungen mit variierendem pH, Übersättigungsgrad und Karbonatgehalt gewonnen wurden, wurden mit physikochemischen und morphologischen Methoden untersucht. Röntgendiffraktion, Infrarot-Spektrophotometrie und Elektronendiffraktionsmessungen bestätigten die amorphe Natur jedes Niederschlages. Sowohl Durchstrahlungs-, als auch Raster-Elektronenmikroskopie zeigten, daß die nativen Elementarpartikel aller 5 synthetischen Produkte von kugeliger Form und unabhängig von den angewandten Präparativ-Techniken waren. Vier der Niederschläge enthielten Kügelchen von der gleichen Größe (200–1200 Å Durchmesser), während jener mit relativ hohem Karbonatgehalt viel größere Partikel aufwies (700–2000 Å Durchmesser). Die Kügelchen waren für den Elektronenstrahl von einheitlicher Dichte, wenn mit der Stromdichte bei niedrigster brauchbarer Stufe betrachtet. Wurden die Proben stärkeren Elektronenströmen ausgesetzt, so entwickelten sich sogleich in jedem Teilchen zahlreiche elektronendurchlässige Zentren. Erhitzen in Vakuum bis zu Temperaturen von 600° veränderte weder die amorphe Beschaffenheit der Niederschläge noch die Morphologie und Größe ihrer elementaren Partikel, obwohl beinahe alles Wasser, welches in den ursprünglich vorliegenden Proben enthalten war, durch dieses Vorgehen entfernt wurde. Die Hitzebehandlung verursachte keine Bildung von elektronendurchlässigen Zentren sondern machte die Kügelchen im Gegenteil in zunehmendem Maße widerstandsfähig gegenüber Strahlenschädigungen. Diese Befunde lassen vermuten, daß sich die ausgestrahlten Elektronen und die Sauerstoff- oder Wasserstoffatome der wäßrigen Komponente des amorphen Calciumphosphates gegenseitig beeinflussen und daß diese anfängliche Reaktion von einer Verdrängung und/oder einem Verlust anderer Atome gefolgt wird.

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Résumé Cinq précipités non-cristallins de phosphate de calcium, préparés à partir de solutions qui varient en pH, degré de saturation et contenu en carbonate, sont examinés par des méthodes physico-chimiques et morphologiques. Des mesures de diffractions électronique et en rayons X et de spectrophotométrie infra-rouge confirment la nature amorphe de chaque précipité. Le microscope électronique par transmission et par balayage démontre que les particules élémentaires des 5 produits synthétiques sont de forme sphérique, quelque soit la méthode de préparation. Quatre des précipités contiennent de petites sphères de taille identique (200 à 1200 Å de diamètre), alors que le dernier précipité, riche en carbonate, contient des particules plus grandes (700 à 2000 Å de diamètre). Ces sphères sont de densité électronique uniforme lorsqu'une intensité de courant, la moins élevée, est utilisée. Un faisceau d'électrons, d'intensité plus élevée, permet de mettre en évidence de nombreux centres, transparents aux électrons, dans chaque particule. In chauffage sous vide à 600° ne modifie pas la nature amorphe des précipités ou la morphologie et la taille des particules élémentaires, bien que la presque totalité de l'eau, contenu dans ces précipités, soit éliminée. L'échauffement ne provoque pas la formation de centres transparents aux électrons. Au contraire, les sphères deviennent résistantes à des dommages, causés par le faisceau. Il semble que le faisceau d'électrons agit sur les atomes d'oxygène et d'hydrogène de la phase aqueuse du phosphate de calcium amorphe et que cette réaction initiale est suivie par le déplacement et/ou la perte d'autres atomes.

     

Nerve endings in the pulmonary trunk, ductus arteriosus and aorta of intact and decapitated pig fetuses

Summary Nerve fibres reactive to acetyl-thiocholine, and tissues showing catecholamine fluorescence were examined in the pulmonary trunk, ductus arteriosus and aorta of 28 pig fetuses between 31 and 113 days of gestation (term=114±1 days). Eight additional fetuses, which had been decapitated in utero at 40–43 days, were also studied at ages between 51 and 114 days of gestation. Spherical micro-networks of nervous tissue reactive to acetyl-thiocholine are present in the adventitia on the cranial aspect of the pulmonary trunk and ductus arteriosus, between the aorta and pulmonary trunk, and on the caudal aspects of the pulmonary trunk and the pulmonary arteries. These fibres invest spherical clusters of catecholamine containing cells which are well supplied with blood vessels. Nerve fibres which fluoresce are also found in association with these cells. Decapitation in utero does not appear to affect the distribution or morphology of these structures. The observations show that structures are present in the major arteries of the fetal pig which may act as sensory receptors, and that these structures are unaffected by chronic vagotomy of the fetus produced by decapitation early in gestation.     

Electron microscopic study of metachromatic leucodystrophy

Summary In metachromatic leucodystrophy abnormal lipids are stored within the lysosomes of the glial cells and phagocytes. The lipid inclusions which are the less structured are the richest in acid phosphatases. Those of which the ultrastructure is prismatic show a very low enzymatic activity and are more similar to residual bodies than to active lysosomes. It appears that the lack in lysosomal arylsulfatase leads to an accumulation of the sulfatide moiety of the stored lipids while the other lipids originating from the myelin breakdown, would be normally catabolized. This leads to a progressive transformation of the ultrastructure of the lysosome contents.

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Résumé Dans la leucodystrophie métachromatique, les lipides anormaux s'accumulent dans les lysosomes des cellules gliales et des phagocytes. Les inclusions lipidiques les moins organisées sont les plus riches en phosphatases acides. Celles dont l'ultrastructure est prismatique ont une activité enzymatique très faible et semblent plus proches de corps résiduels que de lysosomes actifs. Il semble que l'absence d'arylsulfatase lysosomiale induise la persistance des sulfatides accumulés dans les lysosomes alors que les autres lipides provenant de la dégénérescence myélinique, seraient normalement catabolisés. Un remaniement progressif de l'ultrastructure du contenu des lysosomes en est la conséquence.