鸡新城疫副眼和副鼻器官微环境局部免疫机理的研究——Ⅰ.血清学和免疫荧光研究

本文研究了用新城疫B_1系疫苗免疫后的SPF雏鸡的泪液、唾液,气管粘液和血液中的血凝抑制(HI)抗体的性质和动态变化规律;同时采取哈德泪腺、气管、肺脏、脾脏、胸腺和法氏囊,作低温组织切片间接免疫荧光试验。结果证明:泪液、唾液和气管粘液中存在着HI抗体.经过两次免疫后,这种抗体至少可持续71天,而且在免疫初期(泪液在一免后的第26天前,唾液和气管粘液在一免后的第29天前)这些外分泌液中的HI抗体滴度要比血液中相应的抗体滴度高.说明这些抗体主要是由局部免疫组织产生的,而不是由血液中机械性渗透来的。哈德泪腺是以B淋巴细胞为主的外周免疫器官,它是在局部产生抗体的主要部位,同时气管、肺脏的局部淋巴组织也参与抗体的合成。参与局部免疫的抗体主要属于IgA。局部免疫反应以体液免疫为主,并由体液免疫和细胞免疫共同完成。     

人胎气管内胰岛淀粉样多肽免疫反应细胞的个体发生(英文)

为了进一步探讨胰岛淀粉样多肽（ＩＡＰＰ）的分布、定位以及它与其他生物活性物质的关系;用ＩＡＰＰ组织化学ＰＡＰ邻片双标法,观察了１８例１４～３８周人胎气管内ＩＡＰＰ免疫反应（ＩＲ）细胞的个体发生及与５羟色胺（５－ＨＴ）的关系。结果显示,胎１４周,气管粘膜表面的假复层柱状上皮中已有ＩＡＰＰ－ＩＲ细胞（Ｆｉｇ．１＆２）;１５周开始,粘膜固有层气管腺导管上皮中也出现分散的ＩＡＰＰ－ＩＲ细胞（Ｆｉｇ．３）;随胎龄增长,１７～２１周,气管上皮内ＩＡＰＰ－ＩＲ细胞逐渐增多;免疫染色加深（Ｆｉｇ．４＆５）,有些细胞发出细突直达腔面（Ｆｉｇ．６＆７）,粘膜下层的气管腺腺泡中也有ＩＡＰＰ－ＩＲ（Ｆｉｇ．８）; ２２～３８周,气管内 ＩＡＰＰ－ＩＲ 细胞又呈逐渐减少趋势,ＩＡＰＰ－ＩＲ仅出现在基底锥形细胞中（Ｆｉｇ．９＆１０）,且免疫染色较深。邻片未显５－ＨＴ－ＩＲ。本研究表明,人胎儿期气管上皮细胞内有ＩＡＰＰ的表达;且ＩＡＰＰ－ＩＲ细胞随胎期的发育而发生变化。     

小熊猫胃的解剖和组织结构研究

小熊猫的胃属单室腺型胃,它以角切迹为界,可分为胃底部和幽门部两部分。胃壁由粘膜、粘膜下层、肌层和浆膜四层组成。四上皮为单层柱状上皮,具有分泌粘液的功能。胃腺有贲门腺、胃底腺、幽门腺三种,但贲门腺不发达。主细胞、壁细胞和粘液细胞的数量与分布呈现规律性变化。肌层发达,特别是内环行肌发达。并与大熊猫胃的结构作了比较。     

小熊猫胃的解剖和组织结构研究

小熊猫的胃属单室腺型胃,它以角切迹为界,可分为胃底部和幽门部两部分.胃壁由粘膜、粘膜下层、肌层和浆膜四层组成.四上皮为单层柱状上皮,具有分泌粘液的功能.胃腺有贲门腺、胃底腺、幽门腺三种,但贲门腺不发达.主细胞、壁细胞和粘液细胞的数量与分布呈现规律性变化.肌层发达,特别是内环行肌发达.并与大熊猫胃的结构作了比较.     

巴西彩龟消化道组织学的初步观察

巴西彩龟的消化道管壁由内向外依次由粘膜、粘膜下层、肌层和浆膜4层结构组成.食管、胃和大肠有纵行粘膜皱襞,小肠无皱襞但有绒毛.食管上皮和肠上皮为复层柱状上皮,胃上皮为单层柱状上皮.消化道粘膜固有层内分布有淋巴细胞,虽然淋巴细胞有聚集现象,但没有形成典型的淋巴小结.无食管腺和肠腺,虽然有胃腺但胃腺细胞分化不明显,说明巴西彩龟消化道的组织分化程度较低,在系统发育中处于较低等地位.     

人胎大肠氮能神经元发育的研究

用NADPH-d组织化学法对人胎大肠氮能神经元的发育进行了观察.结果表明第5个月胎龄时,肌间神经节处圆形细胞中部分细胞出现一氧化氮合酶(NOS)阳性反应,并分化成氮能神经细胞.第6个月胎龄时,氮能神经元胞体增大,突起伸长,在肌层、粘膜下层和肠腺基部出现氮能神经纤维分布.第7个月胎龄时,氮能神经元生长发育达到高峰,肌间神经节细胞数目增多,环肌层神经纤维分布密度增加,膨体结构明显.第8-10个月胎龄时,氮能神经元染色强度加深,其胞体分布以肌间神经节最多,粘膜下层和内环肌层较少.氮能神经纤维的分布密度以内环肌层最高,粘膜下层和外纵肌层次之,粘膜层较低.本研究揭示了大肠氮能神经元发育的变化规律.     

东方铃蟾消化道组织学的初步研究

采用组织学方法对东方铃蟾的消化道进行了研究。结果表明:肠分为十二指肠、空肠和大肠。消化道管壁由粘膜层、粘膜下层、肌层和浆膜层构成。食道、胃和肠均为单层柱状上皮。胃和十二指肠的粘膜皱褶最丰富。食道腺为复泡状腺,胃腺属于单管状腺,肠的各段无多细胞腺体,但空肠和大肠有丰富的杯状细胞。肌层均为平滑肌,内层环肌较厚,外侧纵肌较薄,其中大肠的外侧纵肌最发达。     

实验性脾虚证大鼠结肠和盲肠组织化学、免疫组织化学研究

成年雄性Wistar大鼠60只,分正常对照组;脾虚组;自然恢复组和四君子汤治疗组。四组动物取结肠和盲肠分别进行H-E;酶组织化学和5-羟色胺细胞PAP免疫组织化学观察。结果表明,脾虚组结肠和盲肠粘膜上皮的杯状细胞较对照组明显增多,淋巴细胞及淋巴集结也增多。固有层毛细血管充血并有炎症细胞的增多。上皮细胞和腺细胞琥珀酸脱氢酶(SDH)减弱,乳酸脱氢酶(LDH)增强;结肠上皮细胞和腺细胞碱性磷酸酶(AIP)和三磷酸腺苷酶(ATPase)增强,而盲肠上皮细胞和腺细胞AIP和ATPase减弱。结肠和盲肠5-HT细胞免疫组化反应减弱。这些结果表明肠上皮细胞酶的变化和5-HT细胞分泌活性的变化与脾虚证的发生密切相关,可能是导致脾虚证原因之一。经四君子汤治疗后以上各项指标均比自然恢复组更接近于正常对照组,说明此药对消化道粘膜上皮酶活性和5-HT细胞分泌活性恢复正常有明显效果,从而起治疗作用。     

石磺消化系统的组织学观察

对石磺消化系统各部分结构进行组织学观察.石磺的消化系统由消化道和消化腺两部分组成.消化道包括口、食道、贲门胃、幽门胃、中肠和后肠,不具吻;消化腺包括肝胰腺、唾液腺和肛门腺.在光学显微镜下,消化道由粘膜层、粘膜下层、肌层和外膜4层组成;肌层主要为环肌,粘膜层主要为柱状细胞.肝胰腺甚为发达,组织结构显示肝胰腺由很多分支的腺管组成,腺管由腺细胞、分泌细胞等组成.唾液腺和肛门腺发达.     

中华花龟消化系统的组织学初步研究

采用常规石蜡切片的方法,对4只成体中华花龟（Oeadia sinensis）的消化系统进行组织学观察。结果表明,中华花龟的消化道管壁除口腔外均由粘膜层、粘膜下层、肌层和浆膜组成;消化道各部分的差别主要在于粘膜层和肌层,舌、咽上皮为复层柱状上皮,食道、胃、肠上皮为单层柱状上皮,大肠上皮为复层扁平上皮;食道粘膜上皮特化成与水呼吸有关的绒毛,胃体部肌层最发达,内斜中环外纵相间排列,厚约652．6±41．2μm,小肠绒毛长而密集呈叶状;肝实质内含大小不等的色素细胞,门管区明显,肝小叶分界不清;胰腺腺泡细胞发达．内分泌细胞零散分布。     

Chicken fetal antigen: association with lymphoid development and differentiation

Rabbit antisera capable of detecting chicken fetal antigen (CFA) was prepared against 1-day chick red blood cells (RBCs) and made specific by exhaustive adsorption with adult chicken peripheral RBCs (PRBCs) from the same strain. Microcytotoxicity was used to study the incidence of CFA on lymphocytes obtained from several organs at different developmental stages in the chicken. Lymphocyte-associated CFA (LA-CFA) determinants and erythrocyte-specific CFA (ES-CFA) determinants were distinguished through the use of adsorption analysis. The high incidence of CFA-positive lymphocytes found in the fetal bursa and thymus was not equaled in the peripheral organs of the spleen, cecal tonsils, and gland of Harder. CFA expression on adult lymphocytes was restricted to the thymus and peripheral blood. It is suggested that these cells may represent a subpopulation of T lymphocytes. Adult spleen, cecal tonsils, and gland of Harder were virtually devoid of CFA-bearing lymphocytes. At fetal developmental stages when greater than 94% of the bursal B cells were CFA-positive, few, if any, of the highly differentiated Harderian B cells possessed CFA. It is suggested that LA-CFA expression is dependent upon B cell differentiation and/or the bursa (central) vs gland of Harder (peripheral) microenvironment. The pattern of CFA expression on bursacytes is discussed in light of the properties of age resistance and bursal-dependent target cells associated with virally induced lymphoid leukosis.     

Effect of emetine on the plasma cell population of chicken's gland of Harder.

The emetine effectively abolished the plasma cell population in the chicken's gland of Harder by day 3 of treatment. The plasma cell content regenerated by day 5 following emetine injection, possibly from a metabolically inactive, resting B cell population which was resistant to the emetine treatment. By day 7 extracellular substance in a very large quantity appeared among the plasma cells and epithelial cells which might represent a hyperactive plasma cell secretion. The changes in the circulating antibodies measured by hemagglutination well-correlated with the plasma cell content in the gland of Harder. The gland of Harder (GH) is an accessory lacrimal gland. Its main function is to lubricate the nictitating membrane and keep the surface of the eyeball wet. The presence and regulatory function of cAMP dependent histone kinase was showed. Since it has been published that in chicken the interstitium of this gland contains a proper amount of plasma cells, this observation called the attention of many investigators to study the role of GH in the immune response. The B cell maturation in the chicken GH has been studied. The surface marker studies have proved that beside the B cells the gland contains functionally adequate number of T cell which exert stimulatory effect on B cells to promote their transformation to plasma cells. In addition to T and B cells small number of macrophages also occur. In the 9 H the number of plasma cells is age dependent. At hatch only a few plasma cells occur in the interstitium of the gland but by 3 weeks of age they become predominant. Different isotypes of immunoglobulins are secreted by these plasma cells.     

Transformed growth phenotype of mouse mammary epithelium in primary culture induced by specific fetal mesenchymes.

When mesenchyme from fetal mammary or salivary gland is implanted into adult mouse mammary gland, adjacent epithelium responds with intense hyperplasia. The hyperplastic cells are more vulnerable than are non-stimulated cells to transformation in vivo by a chemical carcinogen or by mammary tumor virus. This system provides a potentially useful model for determining how stroma contributes to mammary tumorigenesis. We have developed co-culture systems and used them to investigate in more detail the nature of the signal produced by the mesenchyme cells. Monolayers of mesenchyme cells were prepared on tissue-culture wells. The mesenchyme cells were trapped on the surface by a thin overlay of agarose. Primary mammary epithelial cells were cultured atop this barrier layer, either as organoids in collagen gels for assessment of anchorage-dependent growth, or as single-cell dispersions in soft agarose for assessment of anchorage-independent growth. Our procedures for assay of anchorage-independent growth allow us for the first time to detect and measure this transformation-defining characteristic in non-immortalized mammary epithelial cells in primary culture. Fetal mammary fat pad precursor tissue and fetal salivary mesenchyme both stimulated anchorage-dependent growth of mammary epithelium, with cell number increasing as much as fifteenfold during a 6-day culture period. These same fetal tissues also stimulated anchorage-independent growth of the mammary epithelial cells, with colony-forming efficiencies of up to 40% in co-cultures with salivary mesenchyme. No colonies formed in the absence of mesenchyme. Cells of colonies contained keratin, which indicates that the colonies grew from epithelial cells and not from a contaminant of another cell type. When co-cultured epithelial cells were subsequently re-cultured in the absence of mesenchyme, they lost their ability to grow independent of anchorage. No colonies grew in co-cultures with fetal cells from heart, kidney, or lung, which is consistent with the lack of stimulation by these tissues in the mammary gland in vivo. A tumor promoter, 12-O-tetradecanoylphorbol acetate (TPA), also caused anchorage-independent growth of the dispersed mammary epithelial cells. Culture medium conditioned by primary or early-passage salivary mesenchyme cells was capable of stimulating growth under both anchorage-dependent and anchorage-independent conditions, confirming that these effects are mediated by a paracrine factor. The results indicate that stimulatory fetal mesenchymes produce soluble molecules that act analogously to transforming growth factors.     

Effects of mesenchyme on epithelial tissue architecture revealed by tissue recombination experiments between the submandibular gland and lung of embryonic mice

Lung epithelium during morphogenesis maintains a sheet structure of polarized cells lining a lumen, in which E-cadherin, β-catenin and tight junctional proteins are localized at the cell–cell contact sites. On the other hand, the submandibular gland epithelium at early stages of development forms a non-cavitated mass of cells where E-cadherin/β-catenin are present on the entire cell surfaces and tight junctional proteins are almost absent or weakly scattered. In the present study, tissue recombination experiments were performed between the two organs to explore roles of mesenchyme in the architectural development of the epithelium. Homotypic recombinants of both submandibular gland and lung showed the tissue architecture as observed in the intact organs. In contrast, 11-day lung epithelium cultured with 13-day submandibular mesenchyme formed multilayers of cells with the lumen being less visible. It was accompanied by redistribution of E-cadherin/β-catenin along the entire cell surfaces and by an irregular distribution of tight junctional proteins. A similar redistribution of these molecules was observed in 15-day lung epithelium cultured with the submandibular mesenchyme, although the epithelial sheet structure lining the lumen was formed. On the other hand, the tissue architecture of submandibular gland epithelium was little affected by lung mesenchyme, although the epithelium was flattened and showed branching morphogenesis.     

Early Development of Mouse Anterior Pituitary: Role of Mesenchyme

Epithelial-mesenchymal interaction in the early development of the anterior pituitary gland was examined by chronological observations on fetal pituitary epithelium grafted in vivo with and without its own mesenchyme. At 8.5 days of gestation, the RATHKE'S pouch began to evaginate toward the diencephalon. The mesenchymal tissue around the pouch was at first very sparsely scattered, but then condensed, on day 10 becoming visible under a dissecting microscope. When RATHKE'S pouch epithelia from 10- and 12-day fetuses were transplanted alone under the kidney capsule, they proliferated slightly to form cysts, the cells of which differentiated into ACTH-producing cells, but not into prolactin-producing cells. Pituitary morphogenesis did not occur. When these epithelia were recombined with homotypic mesenchyme and transplanted, the epithelia proliferated remarkably on one side of the wall of the pouch, resulting in formation of a pars distalis that contained both ACTH-producing cells and prolactin-producing cells. Heterotypic mesenchyme, such as lung, dermis and mammary gland mesenchyme, could induce 12-day epithelium, but not 10-day epithelium to develop into pars distalis. Thus, fetal pituitary epithelium has the capacity of autodifferentiation into ACTH-producing cells, not into prolactin-producing cells, and requires mesenchymal support for development of the pars distalis.     

Involvement of pro-apoptotic and anti-apoptotic factors in the early development of the human pituitary gland

The spatial and temporal pattern of appearance of pro-apoptotic caspase-3 and p53 proteins, and anti-apoptotic bcl-2 protein was investigated in the developing pituitary gland of 6 human embryos 5-8-weeks old, using morphological and immunohistochemical techniques. Their dynamic appearance was analyzed in the Rathke's pouch (future adenohypophysis), mesenchyme, and in the developing neurohypophysis. In the 5th and 6th week, caspase-3 positive cells appeared in the Rathke's pouch (5%) and stalk (11%), in the mesenchyme, but not in the neurohypophysis. In the 6th and 7th week, apoptotic cells were more numerous in the caudal part of the Rathke's pouch due to its separation from the oral epithelium. Pro-apoptotic p53 protein was detected in all parts of the pituitary gland throughout the investigated period. Nuclear condensations characterized cells positive to caspase-3 and p53 proteins. Apoptotic cells displayed condensations of nuclear chromatin on an ultrastructural level as well. While caspase-3 dependent pathway of cell death participated in morphogenesis of the adenohypophysis and associated connective tissue, p53-mediated apoptosis most likely participates in morphogenesis of all parts of the gland, including neurohypophysis. The anti-apoptotic bcl-2 protein was also detected in all parts of the developing gland. With advancing development, the positivity to bcl-2 protein increased in the cells of the adenohypophysis, while it decreased in the neurohypophysis. Bcl-2 protein probably prevented cell death in all parts of the gland and enhanced cell differentiation. The described pattern of appearance of the investigated pro-apoptotic and anti-apoptotic factors might be important for normal morphogenesis and function of the pituitary gland.     

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.     

Induction of alveolar type II cell differentiation in embryonic tracheal epithelium in mesenchyme-free culture

We have previously shown that fetal lung mesenchyme can reprogram embryonic rat tracheal epithelium to express a distal lung phenotype. We have also demonstrated that embryonic rat lung epithelium can be induced to proliferate and differentiate in the absence of lung mesenchyme. In the present study we used a complex growth medium to induce proliferation and distal lung epithelial differentiation in embryonic tracheal epithelium. Day-13 embryonic rat tracheal epithelium was separated from its mesenchyme, enrobed in growth factor-reduced Matrigel, and cultured for up to 7 days in medium containing charcoal-stripped serum, insulin, epidermal growth factor, hepatocyte growth factor, cholera toxin, fibroblast growth factor 1 (FGF1), and keratinocyte growth factor (FGF7). The tracheal epithelial cells proliferated extensively in this medium, forming lobulated structures within the extracellular matrix. Many of the cells differentiated to express a type II epithelial cell phenotype, as evidenced by expression of SP-C and osmiophilic lamellar bodies. Deletion studies showed that serum, insulin, cholera toxin, and FGF7 were necessary for maximum growth. While no single deletion abrogated expression of SP-C, deleting both FGF7 and FGF1 inhibited growth and prevented SP-C expression. FGF7 or FGF1 as single additions to the medium, however, were unable to induce SP-C expression, which required the additional presence of serum or cholera toxin. FGF10, which binds the same receptor as FGF7, did not support transdifferentiation when used in place of FGF7. These data indicate that FGF7 is necessary, but not sufficient by itself, to induce the distal rat lung epithelial phenotype, and that FGF7 and FGF10 play distinct roles in lung development.     

Bmpr1a signaling plays critical roles in palatal shelf growth and palatal bone formation

Cleft palate, including submucous cleft palate, is among the most common birth defects in humans. While overt cleft palate results from defects in growth or fusion of the developing palatal shelves, submucous cleft palate is characterized by defects in palatal bones. In this report, we show that the Bmpr1a gene, encoding a type I receptor for bone morphogenetic proteins (Bmp), is preferentially expressed in the primary palate and anterior secondary palate during palatal outgrowth. Following palatal fusion, Bmpr1a mRNA expression was upregulated in the condensed mesenchyme progenitors of palatal bone. Tissue-specific inactivation of Bmpr1a in the developing palatal mesenchyme in mice caused reduced cell proliferation in the primary and anterior secondary palate, resulting in partial cleft of the anterior palate at birth. Expression of Msx1 and Fgf10 was downregulated in the anterior palate mesenchyme and expression of Shh was downregulated in the anterior palatal epithelium in the Bmpr1a conditional mutant embryos, indicating that Bmp signaling regulates mesenchymal-epithelial interactions during palatal outgrowth. In addition, formation of the palatal processes of the maxilla was blocked while formation of the palatal processes of the palatine was significantly delayed, resulting in submucous cleft of the hard palate in the mutant mice. Our data indicate that Bmp signaling plays critical roles in the regulation of palatal mesenchyme condensation and osteoblast differentiation during palatal bone formation.