正常大鼠肺内表皮生长因子受体(EGFR)的免疫组化定位

作者以免疫组织化学技术对大鼠肺组织内表皮生长因子受体(EGFR)进行了定位研究.结果显示 EGFR 广泛分布于支气管粘膜上皮细胞、肺泡细胞、血管内皮下结缔组织和血管平滑肌细胞上。其提示 EGF 通过作用于肺组织内的特异性 EGFR 而发挥其生理和/或病理学功能。     

阿霉素肾病大鼠表皮生长因子及其受体的表达

目的研究阿霉素肾病大鼠肾组织中表皮生长因子(EGF)及其受体EGFR的表达分布以及表达量与尿蛋白之间关系。方法选择第5天、14天、28天作为动态观察的时点,同期设立正常对照。采用荧光定量RT-PCR、免疫组织化学及计算机图像定量分析EGF mRNA以及EGF、EGFR蛋白在肾组织的表达,同时测定24 h尿蛋白定量。WT1和EGFR双重免疫组化确定EGFR在肾小球内确切细胞定位。结果阿霉素注射后第5天,EGFmRNA即较正常增高,28 d明显增高并高于5 d和14 d。正常对照组EGF阳性细胞主要分布于远曲小管和髓袢,阿霉素组EGF还在集合管和近曲小管上表达;EGF阳性表达范围和强度随尿蛋白增加而增加;EGFmRNA表达量以及EGF在肾小管中的表达强度与24 h尿蛋白量呈正相关。肾小管上皮细胞广泛表达EGFR,阿霉素组EGFR在小管表达均高于正常,但组间各时点差异无显著性;随尿蛋白增加EGFR在肾小球内表达逐渐增多。EGFR在肾小球和肾小管中的表达强度均与24 h尿蛋白量呈正相关。WT1和EGFR双重免疫组化显示阿霉素肾病组EGFR可在足突细胞上表达,正常组则无。结论阿霉素肾病大鼠的肾小球脏层上皮有EGFR的表达。EGF/EGFR可能参与了阿霉素肾病的发病过程以及蛋白尿的形成。     

骨髓单个核细胞移植对大鼠溃疡性结肠炎的作用

目的:探讨同种异体骨髓单个核细胞(BM-MNCs)移植大鼠溃疡性结肠炎模型的作用。方法:将DAPI标记的同种异体大鼠骨髓单个核细胞(BM-MNCs)经尾静脉注射移植到大鼠溃疡性结肠炎(UC)模型体内(模型组),以尾静脉注射等量PBS的UC大鼠作为对照组。光镜观察大鼠结肠组织病变改变,荧光显微镜观察标记DAPI的BM-MNCs在结肠组织中的定植及分布情况,免疫荧光检测BM-MNCs中CK19、CD34的表达情况。结果:移植组大鼠结肠组织可见新生黏膜上皮及腺体,黏膜下有新鲜至成熟肉芽组织生成,明显优于对照组;移植14天,大鼠结肠组织中可观察到DAPI标记的BM-MNCs细胞;DAPI标记的细胞可表达血管内皮细胞特异性表达蛋白CD34或黏膜上皮细胞特异性表达蛋白CK19。结论:BM-MNCs可向受损病变部位结肠组织迁移和定植,且分化为血管内皮细胞和黏膜上皮细胞。     

扬子鳄胚胎背腺的发生及退化

本文在１４例扬子鳄Ａｌｌｉｇａｔｏｒｓｉｎｅｎｓｉｓ胚胎中观察了背腺的发生及退化过程。孵化第２８天,背中线左右两侧第二行鳞片处的表皮内陷形成实心的背腺腺芽;第３８天,背腺腺泡明显,腺上皮为复层上皮;从第４６天开始,腺上皮出现明显的退化,大量增殖的腺管上皮细胞逐渐堵塞腺管及腺孔。扫描电镜观察表明,孵化第３２—３６天的胚胎背部第二行鳞的各列鳞片表面均有背腺腺孔,以后逐渐出现少数不规则的退化,孵化第５８天以后,绝大多数背腺腺孔消失。对扬子鳄背腺的发生及退化现象作了讨论。     

着床期小鼠子宫内膜中EGF及其受体转录和表达状况的研究

为探讨表皮生长因子（epidermal growth factor,EGF)在胚泡着床过程中的作用。本文应用原位杂交和免疫组织化学方法,检测了EGE及其受体在胚泡着床前后小鼠子宫内膜中的转录和表达。结果显示：未孕和受精后第4-5天,子宫内膜表面上皮和腺上皮细胞仍呈EGF,EGFR原位杂交和免疫组化阴性着色,受精后第4-5天子宫内膜基质细胞EGF及其受体转录和表达较未孕期增强,受精后第6天,EGF及其受体免疫组化和原位杂交阳性着色主要分布于初级蜕膜带（primary decidual zone,PDZ);随着胚泡植入的进行,PDZ区蜕膜细胞EGF及其受体的转录和表达明显减少,而PDZ周围蜕膜细胞EGF及其受体的转录和表达增强,结果提示,EGF是小鼠胚泡着床过程中的一个重要调节因子。     

角质形成细胞生长因子(KGF)在喉粘膜良性、癌前及恶性病变中的mRNA水平分析

利用原位杂交的方法检测KGFmRNA在正常喉粘膜上皮（N）、慢性非特异性炎症（IF）、不典型增生（DYS）及鳞癌（SCC）中的转录水平,探讨KGF在喉粘膜良性及恶性病变中的分布和可能的作用。结果表明,KGFmRNA不仅在间质中的成纤维细胞中表达,少量的炎细胞及血管内皮细胞中亦表达,而且从N、IF、DYS到SCC、KGFmRNA转录水平逐渐增强;上皮细胞及肿瘤性上皮细胞不表达KGFmRNA,KGFmRNA在分化差的SCC周围间质中表达较分化好的SCC周围间质增多。结论：KGF在上皮与间充质细胞的交互作用中发挥着重要的作用,对维持喉粘膜正常结构、代谢及喉癌的发生发展具有重要意义。     

中华大蟾蜍多种组织内5-羟色胺免疫染色细胞的分布

用过氧化物酶-抗过氧化物酶(PAP)法,对中华大蟾蜍消化道(冬眠期与非冬眠期),脑及其他组织的5-HT分布进行了研究。5-HT免疫染色细胞位于脑干中缝核区和间脑的第Ⅲ脑室腹侧的室管膜细胞区。阳性神经元呈圆形或卵圆形,细胞常有突起与其他阳性细胞突起相连,上述部位中还有一些阳性神经纤维。消化道的免疫染色细胞密度在胃幽门、胃体和胃贲门处最高,食道和十二指肠次之,大肠和小肠最低。非冬眠期蟾蜍消化道内免疫染色细胞密度明显高于冬眠期的(P<0.05)。阳性细胞位于粘膜上皮或腺上皮细胞间,细胞有一个或一个以上呈阳性反应的突起,有的突起伸入肠腔面或腺腔面,有的穿过基膜到达固有层,表明这些细胞兼有内、外分泌的功能。在甲状旁腺的主细胞间,肺呼吸性细支气管上皮和肺泡管上皮细胞间都有5-HT免疫染色细胞,细胞呈立方形、圆形、卵圆形或不规则形,常有几个细胞成簇分布。     

表皮生长因子受体在阴道念珠菌病发病过程中的作用初探

目的初步探索表皮生长因子受体(epidermal growth factor receptor,EGFR)在阴道念珠菌病发病过程中的作用及分子机制。方法培养阴道上皮细胞(VK2/E6E7细胞),白念珠菌刺激后利用荧光定量PCR检测EGFR及可能相关的免疫通路因子的表达。构建EGFR-siRNA VK2/E6E7细胞模型,与白念珠菌共培养后,分别利用ELLISA及全自动细胞检测仪检测表皮生长因子受体敲除前后阴道上皮细胞分泌细胞因子的变化以及对白念珠菌感染防御能力的改变。构建阴道念珠菌感染小鼠模型,qPCR检测阴道组织表皮生长因子受体及免疫通路因子的表达;并检测EGFR磷酸化抑制剂阻断通路后,阴道组织局部的真菌载量和炎性细胞的变化。结果 qPCR检测显示,EGFR、STAT3、GM-CSF及IL-1β在VK2/E6E7细胞感染白念珠菌后表达升高,且具有统计学意义(P0.05);ELLISA检测结果显示,EGFR-siRNA VK2/E6E7细胞感染白念珠菌后GM-CSF、IL-8、 IL-1β、 MIP-3α表达显著下降(P0.05);全自动细胞检测仪检测结果显示,EGFR敲除细胞在感染白念珠菌30 h后的防御能力和细胞活性较正常细胞明显降低。qPCR检测显示小鼠阴道感染白念珠菌后EGFR、HER2、STAT3、IL-8表达升高具有统计学意义(P0.05);局部应用磷酸化抑制剂阻断EGFR通路后,小鼠阴道灌洗液菌载量较对照组明显增加,且在感染后第7日差异有统计学意义(P0.05)。结论表皮生长因子受体及其通路因子在阴道念珠菌病发病过程中起到较为重要的作用。     

用BC1抗体检测人正常腺上皮MUC1基因的表达

揭示ＭＵＣ１ 粘蛋白核心肽在人正常腺上皮中的表达模式。用ＢＣ１ 抗体和ＬＡＢ免疫组织化学方法检测组织中的ＭＵＣ１ 核粘蛋白的表达。ＭＵＣ１ 核粘蛋白主要分布于胃粘膜的上皮层和腺颈部细胞,基底部表达少, 呈均质状, 胃窦、胃底的表达无差异。十二指肠、空肠中的表达呈细颗粒状, 弥漫性, 位于核周; 杯状细胞和柱状细胞的表达类似,杯状细胞的粘液滴内未测得ＭＵＣ１ 基因核心肽。宫颈组织中的腺上皮核周及胆囊内存在ＭＵＣ１ 的表达。ＭＵＣ１ 核粘蛋白广泛地存在于人正常腺上皮细胞内, 但具异质性。     

GSK3在人、鼠、猪的肺组织及培养的猪支气管上皮细胞中的表达

目的探讨GSK3在动物肺组织及培养的猪支气管上皮细胞中的表达.方法用免疫组化,免疫细胞荧光法检测GSK3在人、大鼠、小鼠和猪的肺组织及培养的猪支气管上皮细胞中的表达.结果 GSK3α和β广泛表达于人、鼠和猪的肺组织中,定位于胞浆.它们在几种动物肺组织中的表达分布大致相似,主要见于各级支气管上皮细胞、肺泡上皮细胞、粘膜平滑肌细胞和粘膜下腺体.但GSK3α在软骨细胞中表达明显强于GSK3β;几种哺乳动物肺组织中均未检测到GSK3的磷酸化.免疫荧光检测培养的猪支气管上皮细胞中有丰富的GSK3α、β的表达,磷酸化的GSK3信号弱.结论 GSK3α、β丰富表达于不同种属哺乳动物肺组织的支气管上皮、腺体以及肺泡上皮,提示其可能在肺组织的生理功能和病理发生机制中起着重要作用.     

The epithelial origin of lymphocytes in the palatine tonsil of rabbits

Lymphopoiesis was studied by electron microscopy in the palatine tonsil of the rabbit from 18 days gestation to 5 days after birth. At 18 days tonsils formed as mounds of mesenchyma covered with epithelium. At 19 days the basal epithelial cells started to increase in number, eventually forming 'buds' which projected into the mesenchyme. Simultaneously, lymphocytes appeared nearthe epithelium or buds. There was marked resemblance between the basal epithelial cells and the lymphocytes. Budding slowed down after the 25th day, but individual basal cells continued to migrate into the mesenchyme and lymphocytes increased in number. Ultrastructure wassimilar in both types of cells, and differentfrom mesenchymal cells. At 29 days lymphocytes were found in the basal epithelial layer behind an intact basement membrane. The evidence indicated that lymphocytes were derived from epithelium.     

MATURATION OF THE RAT FETAL THYROID

Maturation of the rat fetal thyroid was studied with the aid of I¹³¹ and of fluorescence and electron microscopy. The I¹³¹ concentration of the fetal gland increased exponentially from day 17 to day 20 of gestation and was related to the weight of the fetus (and presumably the weight of the thyroid) and also to the quantity of I¹³¹ accumulated by the fetus. In the 17-day gland, thyroglobulin or immunologically similar material was sparsely present in the incipient lumens of some cell clusters. With maturation, this material increased and was also observed within follicular cells on days 18 to 19 of gestation. On day 20, the specifically reacting material was present in the follicular lumens and was absent from the cytoplasm of follicular epithelium. Ultrastructurally, the earliest thyroid cells examined were replete with all the organelles found in the more mature epithelium. No direct correlation could be made between the cytoplasmic structures and the presence of thyroglobulin, although the granular endoplasmic reticulum was most likely the organelle responsible for synthesis of thyroglobulin. Thyroglobulin or a precursor was found in fetal thyroid cells before measurable quantities of I¹³¹ were concentrated and before cytoplasmic droplets appeared.     

Intracellular accumulation of basement membrane components during morphogenesis of rat submandibular gland

The distribution of two basement membrane (BM) components, laminin (LN) and type IV collagen (COLL IV), during acino-tubular morphogenesis of rat submandibular gland was examined immunohistochemically to determine the role of BM in the development of acino-tubular structures. On day 14 of gestation, LN could be found only in the BM separating an undifferentiated cell cluster of gland epithelium from surrounding mesenchyme. However, during a short period through days 15 to 17, LN was detected not only in the BM but also in intracellular vesicles of the cells of the terminal cluster. Immunoelectron microscopy showed the intracellular immunoreactive sites to be rough endoplasmic reticulum, indicating that active LN synthesis occurs in the cells of the terminal cluster. Intracellular immunostaining of LN disappeared completely on day 19 with the development of simple epithelium from the cell cluster, even though BM remained reactive. COLL IV also was accumulated in the intracellular vesicles of terminal cluster cells on day 16 of gestation but not on day 19. These results indicate that synthesis of certain BM components is transiently stimulated in gland epithelium before the formation of simple epithelial structure, and that these components are significantly involved in morphogenesis of the submandibular gland.     

Early weaning accelerates the differentiation of mucous neck cells in rat gastric mucosa: Possible role of TGFα/EGFR

The development of the gastric mucosa is controlled by hormones, growth factors and feeding behavior. Early weaning (EW), which means the abrupt interruption of suckling, increases proliferation and differentiation in the rat gastric epithelium. Transforming growth factor α (TGFα) is secreted in the stomach, binds to the epidermal growth factor receptor (EGFR) and may control cell proliferation, differentiation and migration. Here, we investigated the influence of suckling–weaning transition on the differentiation of mucous neck cells in the stomach and its association to the expression of TGFα and EGFR. Fifteen-day-old Wistar rats were divided into two groups: suckling (control), in which pups were kept with the dam, and early weaning (EW), in which rats were separated from their mother and fed with hydrated powdered chow. TGFα and EGFR levels were increased at 18 days in EW animals compared to control ones (p<0.05). Histochemical reactions with Periodic Acid-Schiff reagent+Alcian Blue or Bandeiraea simplicifolia II lectin were used to stain the mucous neck cells and showed an increase in this cell population throughout EW, which was more pronounced at 17 days when compared to suckling pups (p<0.05). These morphological results were confirmed by RT-PCR for mucin 6. The levels of mucin 6 mRNA were higher in EW animals from the 16th to the 18th day (1–3 days post-weaning) when compared to the respective control group. Inhibition of EGFR through AG1478 administration to EW animals prevented the expansion of mucous neck cell population induced by EW (p<0.05). Therefore, early weaning up regulated TGFα/EGFR expression and induced differentiation of mucous neck cells. Moreover, we showed that EGFR takes part in the maturation of this cell population. We conclude that regular suckling–weaning transition is crucial to guarantee the development of the gastric mucosa.     

Immunolocalization patterns of cytokeratins during salivary acinar cell development in mice

Embryonic development of the mouse salivary glands begins with epithelial thickening and continues with sequential changes from the pre-bud to terminal bud stages. After birth, morphogenesis proceeds, and the glands develop into a highly branched epithelial structure that terminates with saliva-producing acinar cells at the adult stage. Acinar cells derived from the epithelium are differentiated into serous, mucous, and seromucous types. During differentiation, cytokeratins, intermediate filaments found in most epithelial cells, play vital roles. Although the localization patterns and developmental roles of cytokeratins in different epithelial organs, including the mammary glands, circumvallate papilla, and sweat glands, have been well studied, their stage-specific localization and morphogenetic roles during salivary gland development have yet to be elucidated. Therefore, the aim of this study was to determine the stage and acinar cell type-specific localization pattern of cytokeratins 4, 5, 7, 8, 13, 14, 18, and 19 in the major salivary glands (submandibular, sublingual, and parotid glands) of the mouse at the E15.5, PN0, PN10, and adult stages. In addition, cell physiology, including cell proliferation, was examined during development via immunostaining for Ki67 to understand the cellular mechanisms that govern acinar cell differentiation during salivary gland morphogenesis. The distinct localization patterns of cytokeratins in conjunction with cell physiology will reveal the roles of epithelial cells in salivary gland formation during the differentiation of serous, mucous or seromucous salivary glands.     

Structural changes in the luminal epithelium of the porcine uterus between days 10 and 19 of the estrous cycle

Light and electron microscopy were used to study the morphology of uterine luminal epithelium from 4 or 5 gilts slaughtered on each of days 10, 13, 16, and 19 of the estrous cycle. Ultrastructural evidence indicated that metabolic activity and accumulation of glycogen by the uterine epithelium increased between days 10 and 16 of the estrous cycle. This phase of high synthetic activity had been terminated by day 19, as evidenced by the reduction or absence of glycogen deposits and decreased incidence of organelles associated with synthetic activity. Diffuse degeneration of epithelial cells occurred throughout the period of study but was maximal between days 16 and 19. Mitotic activity indicated that cell replacement also occurred between day 16 and day 19.     

Immunohistochemical studies on the development of avian embryo pituitary corticotrophs under normal and experimental conditions

Summary Immunofluorescent ACTH cells are present in the developing chick pituitary gland from the 9th day of incubation.Rathke pouch grafts from 4–5 day or 5.5 and 6.5 day-old chicks, grafted into chick chorioallantoic membrane and grown for 12 days, gave rise to tinctorially normal pituitary glands in both cases.The early grafts were of pouch epithelium alone, separated from mesenchyme by trypsinization. The later grafts were surrounded by their attached mesenchyme, from which they are virtually inseparable.In 17 out of 18 of the 4.5 day grafts no immunofluorescent ACTH cells developed. (In the 18th case a few feebly stained single cells). In 16 out of 30 of the 5.5 and 6.5 day grafts ACTH cells were present in normal numbers.Of the 3 hypotheses put forward to explain these findings only one appears valid. This is that the ACTH cells are contributed directly by the mesectoderm (neural crest) surrounding the 5.5 and 6.5 day pituitary primordia.     

Possible functional involvement of thymosin beta 4 in developing tooth germ of mouse lower first molar

We examined the detailed in situ expression pattern of thymosin beta 4 (Tβ4) in the developing mouse mandibular first molar. Tβ4 mRNA was expressed in the presumptive dental epithelium at embryonic day 10.5 (E10.5) and in the thickened dental epithelium at E12. An in situ signal was observed in the invaginated epithelial bud at E13, in the enamel organ at E14 and E14.5, and in the primary enamel knot (PEK) at E14.5. The signal was localized in the epithelial cells of the outer layer of the enamel organ at E15 and E15.5. No signal was found in the PEK at these stages. Tβ4 mRNA was expressed in the inner enamel epithelium, cervical loop and dental lamina at E16 and E17. The expression of Tβ4 mRNA was observed in the polarized inner epithelial cells at E18, newborn day 1 (N1) and N2. However, the signal intensity decreased markedly at N3. We herein report for the first time that Tβ4 is distinctly expressed in developing tooth germ, and it may also play functional roles in the initiation, growth and differentiation of tooth germ.