大肠侧向发育型肿瘤细胞株的建立及其鉴定

目的 为了深入研究大肠侧向发育型肿瘤(LST)细胞的生物学特性及其发生发展的机制。方法采用人LST组织进行肿瘤细胞建株及细胞分离与原代培养,传代至70代进行细胞鉴定．包括细胞的形态结构分析、染色体分析和免疫组化分析等,并与非LST细胞作对比。结果 LST细胞为上皮来源的肿瘤细胞,多形性上皮细胞占大多数,其有近似肿瘤细胞的生长特性,群体倍增时间为36h;染色体数量为42-66,85％在三倍体范围内,并具有异常染色体核型;免疫组织化学观察ESA、CK20均呈阳性;超微结构具肿瘤细胞样表现和特征。结论成功建立了1个LST细胞株,并命名为LST-R1细胞,为进一步深入研究LST细胞的生物学特性及大肠癌的发生发展机制奠定了基础。     

''Metaplastic lesions'' in intrahepatic bile ducts in hepatolithiasis: A histochemical and immunohistochemical study

Pathology of the intrahepatic bile ducts bearing calculi was examined with an emphasis on metaplasia in 22 cases of hepatolithiasis and in seven cases of normal livers. Normal livers contained few glandular elements within the bile duct walls and no metaplastic lesions or endocrine cells. In hepatolithiasis, a number of mucous glands resembling pyloric glands (pseudopyloric gland metaplasia) were seen within duct walls in all cases. The epithelial cells of the glands were positive for class III mucin with paradoxical concanavalin A staining which is known to be specific for pyloric glands. These cells were also positive for neutral, sialo- and sulfomucin to a variable extent. Intestinal metaplasia, including goblet cell and Paneth cell metaplasia, was found within duct walls and in covering epithelia in five (23%) cases. Endocrine cells, including argyrophil, argentaffin and gut hormone-containing cells were present in these metaplastic lesions in 13 (59%) cases. The occurrence of endocrine cells was closely associated with intestinal metaplasia, although there were a few endocrine cells in metaplastic pseudopyloric glands. These findings suggest that metaplastic lesions similar to the well-known metaplastic lesions in the gallbladder occur in the intrahepatic bile duct walls in hepatolithiasis. The appearance of metaplastic lesions and endocrine cells may be causally related to chronic inflammatory processes associated with hepatolithiasis.     

白萝卜提取物促胃动力作用中Cajal间质细胞与P物质的变化

目的探讨白萝卜提取物（raphanus sativus 1 extract，Ecr）通过Cajal间质细胞促胃肠动力作用机制。方法40只SD大鼠随机分为A（Ecr）组、B（0．9％氯化钠对照）组分别灌服等体积Ecr和0．9％氯化钠，1h后用双重免疫荧光组织化学标记的方法观察胃电起搏区Cajal间质细胞（interstitial cells of Cajal，ICC）与P物质（SP）阳性神经及产物表达的变化。结果肌间神经丛中ICC与SP阳性神经呈网络样分布，二者紧密毗邻；肌间神经丛及肌层内与ICC结合的SP阳性神经产物表达明显增加（P〈0．01）。结论白萝卜提取物的促胃肠动力作用可能与胃体ICC上结合的SP阳性神经产物表达增高有密切关系。     

Expression of vascular endothelial growth factor receptor‐3 mRNA in the rat developing forebrain and retina

Vascular endothelial growth factor receptor (VEGFR)‐3, a receptor for VEGF‐C and VEGF‐D, is expressed in neural progenitor cells, but there has been no comprehensive study of its distribution in the developing brain. Here, the temporal and cell‐specific expression of VEGFR‐3 mRNA was studied in the developing rat forebrain and eye. Expression appeared along the ventricular and subventricular zones of the lateral and third ventricles showing ongoing neurogenesis as early as embryonic day 13 but was progressively down‐regulated during development and remained in the subventricular zone and rostral migratory stream of the adult forebrain. VEGFR‐3 expression was also detectable in some differentiating and postmitotic neurons in the developing cerebral cortex, including Cajal‐Retzius cells, cortical plate neurons, and subplate neurons. Expression in the subplate increased significantly during the early postnatal period but was absent by postnatal day 14. It was also highly expressed in nonneural tissues of the eye during development, including the retinal pigment epithelium, the retinal ciliary margin, and the lens, but persisted in a subset of cells in the pigmented ciliary epithelium of the adult eye. In contrast, there was weak or undetectable expression in the early neural retina, but a subset of retinal neurons in the postnatal and mature retina showed intense signals. These unique spatiotemporal mRNA expression patterns suggest that VEGFR‐3 might mediate the regulation of both neurogenesis and adult neuronal function in the rat forebrain and eye. J. Comp. Neurol. 518:1064–1081, 2010. © 2009 Wiley‐Liss, Inc.     

失血性休克大鼠中脑导水管周围灰质腹外侧区一氧化氮合酶阳性神经元的变化

目的 观察失血性休克大鼠中脑导水管周围灰质（PAG）腹外侧区的一氧化氮合酶（NOS）阳性神经元的变化。方法 取雄性SD大鼠20只随机分成4组,即对照组（假手术组）、休克时组、休克45 min组和休克90 min组。对照组仅进行左股动脉插管而不放血;其他各组经左股动脉插管放血复制失血性休克模型。分别在休克时、休克45 min和休克90 min灌注固定后取脑干进行冰冻切片,行NADPH-d组化染色观察,并对各组PAG腹外侧区尾段的NOS阳性神经元和其背内侧NOS阳性突起密度进行比较。结果 PAG腹外侧区尾段的NOS阳性神经元密度的变化无统计学意义（P〉0.01）,但在失血性休克后NOS阳性神经元染色明显加深,其背内侧突起增密,变化有统计学意义（P〈0.01）。结论 大鼠PAG腹外侧区的NOS阳性神经元可能参与了失血性休克的发生发展,并且发挥着影响。     

Zellweger Syndrome: A Histochemical Diagnosis of Two Cases

Until 17 years ago the diagnosis of the cerebrohepatorenal Zellweger syndrome (ZS) rested largely on clinical grounds confirmed by pathologic findings at autopsy. The observation that peroxisomes are not detectable morphologically or histochemically in liver or kidney of patients with this syndrome gave histopathologists the opportunity to make the diagnosis of this complex syndrome at biopsy. Catalase reaction of the peroxisomes can be used as a rapid and accurate method to differentiate between ZS and other clinical conditions in which the peroxisomes are present in normal or reduced number. We describe two patients in whom the diagnosis of ZS was made by the absence of histochemical staining for catalase in a liver biopsy. The findings were subsequently confirmed using standard biochemical tests.     

Keratinization of middle ear cholesteatomas

Summary A histochemical study was performed to determine the involvement of epidermal transglutaminase (ETgase) in the keratinization of middle ear cholesteatomatous lesions, and to compare it with its role in the middle ear mucosa and epidermis. In a first assay, we localized the (E)Tgase activity in situ. A second immunohistochemical assay revealed the distribution of the particulate form of ETgase, which is involved in cross-linked envelope formation. A remarkable difference between strongly keratinized epidermal tissues and the cholesteatoma matrix is the frequent observation in the latter of the remnants of (E)Tgase activity in cytosol, even in advanced stages of differentiation. As a consequence, the cell-membrane-associated ETgase activity, and thus the extent of cross-linking within the envelope, is at a lower level than expected. This aspect is reminiscent of the keratinization phenomenon manifested by thin epidermal tissues. In addition, our findings are the first to show that ETgase is a substantial marker of middle ear mucosa.This paper is dedicated to the memory of Prof. E. Gillis, deceased 3 August 1988, aged 51. His guidance will be sorely missed by those who knew and worked with him.     

大鼠腹膜间皮瘤的酶组织化学及超微结构观察

对２２例大鼠腹膜间皮瘤进行酶组织化学和超微结构的综合研究，间皮瘤主要由ＥＣ、ＦＬＣ、ＩＣ、ＰＭＣ细胞组成，上皮瘤细胞脱氢酶活性高于纤维型，脱水酶活性低于纤维型。不同类型间皮瘤细胞酶活性特点表现各异，并与邻近组织或细胞的酶活性改变有一定的相关性。故认为间皮瘤的发生可能为多起源。同时首次报道未定型间皮瘤酶活性及超微结构的特点，这有助于进一步认识间皮瘤的组织发生及鉴别诊断。     

Effect of toxaphene on isolated hepatocytes of the yellowtail flounder, Pleuronectes ferrugineus storer

The histochemical and enzyme cytochemical effects of Toxaphene were investigated using isolated hepatocytes in suspension culture from laboratory-bred juvenile, female yellowtail flounder (Pleuronectes ferrugineus). Hepatocytes were kept in suspension culture for 4 days and exposed for 3 days to a control medium, to a medium with hexane (the solvent of Toxaphene), or to a medium with Toxaphene in two different concentrations (1 and 10 mocrog/ml). Subsequently, the cultivated cells were examined histochemically (Sudan black B, oil red O, Schmorl's reaction) and enzyme cytochemically (acid phosphatase, NADPH-ferrohemoprotein reductase). Toxaphene decreased the viability of the isolated cells significantly, as compared to the control suspensions. Toxaphene also increased the storage of total and neutral lipids (as demonstrated by Sudan black B and oil red O, respectively) in a dose-dependent manner. In addition, Toxaphene increased the enzymatic activity of acid phosphatase, and increased the storage of lipofuscin pigment (as demonstrated by the Schmorl's reaction) within the hepatocytes, suggesting an increase in the number and/or size of the lysosomes. Hexane did not have a significant toxic effect on the isolated hepatocytes. It is concluded that Toxaphene is potentially toxic to fish in a marine environment and that this in vitro system may provide a model for assessing the direct effect of various toxicants on fish hepatocytes.