人胚中肾发育和生长因子及其受体的表达

目的:观察第3~8周人胚中肾的发育和生长因子及其受体的表达。方法:第3~8周人胚,H-E和免疫组织化学染色,光镜观察。结果:人胚中肾于第3周末出现,并从头尾方向同时进行生长和退化过程。中肾于第8周退化。发育第5周中肾的肾小管和肾小球细胞强表达碱性成纤维细胞生长因子(bFGF)和转化生长因子β1(TGF-β1)及其受体。中肾内有核血细胞于第4周出现,第7周数量达到高峰。表皮生长因子、胰岛素样生长因子I、bFGF和TGF-β1及其受体在发育第5~8周中肾的有核血细胞表达。结论:人胚中肾的发育主要在第3~7周。第6~7周人胚中肾内的有核血细胞数量达高峰。人胚中肾内的有核血细胞、肾小管内皮细胞、肾小囊和肾小球细胞表达多种生长因子。     

The expression of fibroblast growth factors and their receptors in Hodgkin's lymphoma

The expression of fibroblast growth factors (FGF1 and FGF2) and their receptors has been reported in a variety of human neoplasms, including haematological neoplasia. Fibroblast growth factors can promote tumour growth directly, or indirectly through promoting the growth of vessels. In the present study, we evaluated the expression of FGF1 and FGF2 as well as FGF receptors 1-4 (FGFR1-FGFR4) in 39 cases of Hodgkin's lymphoma, including all subtypes, as well as Hodgkin's lymphoma cell lines. FGF1 and FGF2 and their receptors FGFR2-FGFR4, but not FGFR1, were found to be expressed by the malignant cells in the majority of cases. Interestingly, only FGFR3, but none of the FGFs or the other FGFRs, was expressed by the Hodgkin's lymphoma cell lines. This indicates that only FGFR3 is constitutively expressed by Hodgkin's lymphoma cells, whereas FGFs and the other FGFRs are induced in vivo. Culture of the Hodgkin's cell lines under conditions of hypoxic stress could induce vascular endothelial growth factor (VEGF) but not FGF expression. FGFs, in contrast to VEGF, might be expressed in response to paracrine stimuli. In situ hybridization did not reveal FGFR3 gene amplification or translocation as the cause of constitutive FGFR3 expression, as has been shown in a subset of multiple myeloma. FGFR3 might be expressed as part of the Hodgkin's cell phenotype. The demonstration of widespread expression of FGFs and some of their receptors in Hodgkin's lymphoma suggests that FGFs are important for sustaining growth of the lymphoma and suggests that anti-FGF antibodies could be used as an adjuvant treatment.     

胎盘生长因子在妊娠期高血压疾病胎盘组织中的定位及定量分析

目的探讨胎盘生长因子(placental growth factor,PLGF)在妊娠期高血压疾病胎盘中定位及定量表达。方法选择妊娠期高血压疾病患者46例,其中子痫前期重度23例,子痫1例;慢性高血压并发子痫前期1例,选择同期正常妊娠妇女20例作为对照组。采用免疫组织化学染色法和免疫印迹(Western blot)方法检测两组患者胎盘PLC蛋白定位及定量表达。结果 (1)免疫组织化学染色发现PLGF蛋白在妊娠期高血压疾病组及正常妊娠组胎盘中分布范围基本一致, 主要分布在绒毛合体滋养细胞和间质细胞的胞浆,部分血管合体膜上也有PLGF阳性染色。(2)Western blot方法检测妊娠期高血压疾病组子痫前期轻、重度胎盘绒毛PLGF蛋白表达低于正常妊娠组(0.3±0.4 vs 0．6±0．4、0．2±0．5 vs 0．6± 0．4),差异有统计学意义(P<0．01);妊娠期高血压患者胎盘中PLGF蛋白的表迭为0．5±0．6,与对照组比较,差异无显著性(P>0．05)。结论胎盘PLGF蛋白表达异常在妊娠期高血压疾病发病中可能具有重要的作用。     

Differential expression of laminin chains and their integrin receptors in human gastric mucosa.

The proliferating cells of the gastric mucosa are found among the pit and mucous neck cells. These cells migrate upward to renew the surface epithelium and downward to restitute the glandular cells. As the epithelial basement membranes (BMs) function as substrate for cell adhesion and migration as well as signals for their differentiation, we studied, by indirect immunofluorescence microscopy, the distribution of different laminin chains and their integrin receptors in adult human stomach. The immunoreactivity for laminin alpha 2 chain localized to the BMs of glands and the lower parts of the gastric pits whereas the laminin alpha 3 chain (laminin-5/kalinin) immunoreactivity was strictly confined to BMs underneath the surface epithelium and the upper parts of the pits. Proliferating mucosal epithelial cells, identified by Ki-67 antibodies, were confined to the areas containing both alpha 2 and alpha 3 laminin chains. The alpha 1, beta 1, and gamma 1 laminin chains were found in all BMs of the mucosa whereas the beta 2 chain was prominent in mucosal blood vessels and also detectable in some glands. Among the laminin integrin receptors, the alpha 3 and beta 4 subunits were seen to be expressed in cells along the BMs with the alpha 3 laminin chain. The alpha 6 integrin, on the other hand, was seen in all gastric epithelia. The present results demonstrate that in the adult human stomach laminin alpha 2 and alpha 3 chains show zonal distribution in BM underlying gastric mucosal epithelium whereas other laminin chains show a more general distribution.     

Toll-like receptor-2 expression in normal and pathologic human placenta

Toll-like receptors (TLRs) are important components of the innate immune system and are expressed by trophoblast in normal full-term placenta. At present, not much is known about the role of TLRs during normal pregnancy and in pregnancy complicated by infection. In this study, we have used immunohistochemistry to investigate the expression of TLR2 in 58 placentas from second and third trimester with chorioamnionitis and 25 full-term placentas from uncomplicated pregnancies without chorioamnionitis. TLR2 was found to be localized to the cyto- and syncytio-trophoblast cell layer and to decidual stromal cells. The expression of TLR2 in placentas with chorioamnionitis was significantly lower than in placentas without chorioamnionitis. Furthermore, there was a significantly higher expression in placentas from liveborn children than in placentas from stillborn/aborted fetuses and also a higher expression in second- than in third-trimester placentas. These data suggest that TLR2 expression in the trophoblast could be involved in the response to infectious pathogens in the placenta.     

Immunocytochemical localization of growth factors and their receptors in human pre-embryos and Fallopian tubes

We utilized indirect immunocytochemistry to demonstrate thepresence of growth factors and their receptors in human pre-embryosand Fallopian tubes. In pre-embryos, only transforming growthfactor- (TGF-) and the intracellu-lar domain of epidermal growthfactor receptor (EGFR) were found at the 4-cell stage. In 8-to 14-cell pre-embryos, TGF-, the intracellular and extracellulardomains of EGFR, and insulin-like growth factor-I and its receptorwere found. Antibodies against TGF- stained all Fallopian tubespecimens, while the extracellular domain of EGFR was only foundin specimens from patients with either blood type A or AB. Theseresults suggest a cross-reactivity between the extracellulardomain of the EGFR and blood group antigens. Our novel demonstrationof growth factor receptor staining in human pre-embryos showsthat growth factor receptor localization is dependent on thedevelopmental stage of human pre-embryos. We have also establisheda potentially important link between the Fallopian tube whichsecretes growth factors and the localization of growth factorreceptors in pre-embryos. These findings are compatible withthe hypothesis that tubal secretions are embryotrophic for theearly development of the pre-embryo.     

Expression of the angiogenic growth factors VEGF, FGF-2, EGF and their receptors in normal human endometrium during the menstrual cycle

Angiogenesis is an important but poorly understood process of the cycling endometrium. Endometrial angiogenesis is believed to be regulated by angiogenic growth factors under the influence of ovarian steroids. Vascular endothelial growth factor (VEGF) and its receptors VEGFR-1 and VEGFR-2, fibroblast growth factor 2 (FGF-2) and its receptors FGFR-1 and FGFR-2, as well as epidermal growth factor (EGF) and its receptor EGFR are believed to be important in the control of angiogenesis in the human endometrium. Their expression was examined by immunohistochemistry in endometrial biopsies obtained from 16 healthy women with proven fertility. Western blot analysis showed that the primary antibodies used were specific for their epitopes. We found that VEGF, FGF-2, EGF and their receptors were all expressed, especially in and/or around blood vessels, thus supporting the hypothesis that these peptides contribute to the regulation of angiogenesis and blood vessel function in the human endometrium. The receptors VEGFR-1, VEGFR-2, FGFR-2 and EGFR were co-expressed and exhibited their strongest expression during the beginning of the secretory phase, coinciding with the developing endometrial oedema and formation of a complex subepithelial capillary plexus. No correlation was seen between receptor expression and stromal blood vessel density.     

p53在妊娠滋养细胞和肿瘤中的表达

目的：分析Ｐ５３过度表达与葡萄胎、恶性葡萄胎（恶葡）、绒毛膜癌（绒癌）的恶性程度及预后关系。方法：采用单克隆抗体免疫组化技术，对正常胎盘绒志及妊娠滋养细胞肿瘤组织，进行肿瘤抑制基因Ｐ５３表达检测。其中包括早期妊娠１０例，中期妊娠１０例，晚期妊娠１３例，葡萄胎４０例，恶葡９例，绒癌２０例；结果：正常胎盘Ｐ５３异常表达阴性，葡萄胎、恶葡、绒癌阳性表达率分别为４２．５％（１７／４０）、５５．５％（５／９     

Expression and localization of placenta growth factor and PlGF receptors in human meningiomas

It has previously been suggested that in human brain tumours, endothelial cell proliferation during angiogenesis is regulated by a paracrine mechanism involving vascular endothelial growth factor (VEGF) and its receptors (VEGF receptor 1 and VEGF receptor 2). The mechanism of growth factor up-regulation is based on hypoxic activation of mRNA expression and mRNA stabilization and genetic events, leading to an increase of growth factor gene expression. The role of the other newly discovered VEGF family members with a high specificity for endothelial cells in the pathogenesis of glial neoplasms is unknown. To investigate which other members of the VEGF family are overexpressed in human brain tumours, the mRNA levels of placenta growth factor (PlGF), VEGF-A, and VEGF-B genes were determined by northern blot analysis in surgically obtained human meningiomas. In the 16 meningiomas examined, the mRNA for PlGF was highly expressed in four tumours and VEGF-A mRNA was highly abundant in three tumour samples. There was no close correlation between PlGF mRNA levels and VEGF-A expression levels. VEGF-B mRNA was abundantly expressed in all tumour samples at uniform levels. In a PlGF-positive tumour sample, immunoreactive VEGFR-1 and VEGFR-2 were detected in endothelial cells of the blood vessels. PlGF protein was detectable in most but not all capillaries of the tumour. PlGF is thus highly up-regulated in a subset of human meningiomas and may therefore have functions, in some tumour vessels, connected to endothelial cell maturation and tube formation. These findings suggest that PlGF, in addition to VEGF-A, may be another positive factor in tumour angiogenesis in human meningiomas. Copyright © 1999 John Wiley & Sons, Ltd.     

Differential expression of high-affinity melatonin receptors (MT1) in normal and malignant human breast tissue

Melatonin is a pineal hormone that strongly inhibits the growth of breast cancer cells in vitro and in vivo. We report thefirst use of immunohistochemical analysis to determine the distribution of the high-affinity melatonin receptor subtype, MTI, in human breast tissue, the hypothalamic suprachiasmatic nucleus, and skin. The MT1 antibody, which is specific for the cytoplasmic portion of the receptor, produced cytoplasmic staining in normal-appearing breast epithelial cells and ductal carcinoma cells; stromal cells, myoepithelial cells, and adipocytes were nonreactive. The majority of nonneoplastic samples (13/19 [68%]) were negative to weakly positive, while moderate to strong reactivity was seen in most cancer samples (49/65 [75%]). Thus, although MT1 receptors were detectable in normal and malignant breast epithelium, high receptor levels occurred more frequently in tumor cells (P < .001), and tumors with moderate or strong reactivity were more likely to be high nuclear grade (P < .045). These findings may have implications for the use of melatonin in breast cancer therapy.     

Differential expression of N-cadherin and E-cadherin in normal human tissues

E-cadherin, which expressed in various epithelial tissues, is important for the maintenance of normal epithelial phenotypes. However, the distribution of N-cadherin in normal human tissues has not been defined systemically. In the present study, we employed a sensitive, reliable immunohistochemical detection system for N-cadherin on formalin-fixed, paraffin-embedded tissue sections, and succeeded in demonstrating N- and E-cadherin protein expressions and their distribution in normal human tissues. E-cadherin immunoreactivity was detected in all the epithelial tissues examined, except for the adrenal cortical cells and granulosa cells. N-cadherin was selectively expressed on epithelial cells of the thymus, pituitary, pancreas, liver, adrenal, endometrium of the uterus, ovary, and stomach as well as in neuronal tissues. Double immunostaining revealed that N-cadherin expression was closely associated with the hormone-producing ability of cells in the pancreas and pituitary. Thus, this study indicated the possibility that N-cadherin is selectively expressed in relation to hormonal regulation in some organs and plays different functions in different situations. The method presented here should prove useful for the further investigation of the N-cadherin expression and function in several disease conditions on formalin-fixed, paraffin-embedded archival tissues.     

转化生长因子3β在妊高征和正常妊娠胎盘组织中的差异表达

目的探讨转化生长因子3β(TGF-3β)蛋白及mRNA在妊高征和正常妊娠胎盘组织中表达的差异.方法取10例妊高征和10例年龄配对的正常妊娠的胎盘组织,应用Western blot方法,检测妊高征和正常妊娠胎盘组织中TGF-3β蛋白的表达水平;应用定量PCR方法,检测妊高征和正常妊娠胎盘组织中TGF-3βmRNA表达水平.结果与正常妊娠胎盘组织TGF-3β蛋白和mRNA表达水平相比,妊高征胎盘组织中的TGF-3βmRNA表达水平增加,妊高征胎盘组织中的TGF-3β蛋白表达水平也明显增加.结论 TGF-3β在妊高征胎盘组织中表达增加,妊高征胎盘中TGF-3β高表达可能与妊高征的病因有关.     

血管生长相关因子在脑血管畸形中的差异性表达

背景：脑血管畸形是在青壮年人群中造成出血性脑卒中的常见病因,畸形血管破裂出血可引发严重的神经功能障碍。脑血管畸形的形成及发病原理尚不明确。现代分子生物学的研究表明血管生长相关因子在脑血管畸形中可能存在异常表达。目的：评价血管生长相关因子在脑畸形血管中的表达差异,探讨血管生长相关因子与脑畸形血管形成的关系。方法：选择脑血管畸形患者与颅内出血开颅治疗患者各50例,通过免疫组织化学染色方法检测脑血管畸形标本和开颅治疗患者颞浅动脉标本中血管因子的表达差异。结果与结论：正常颞浅动脉中血管生长相关因子(血管内皮生长因子和转化生长因子α)几乎不表达,畸形血管中两者高表达(P < 0.05)。结果证实,与正常血管相比,脑血管畸形患者血管内皮生长因子和转化生长因子α的表达存在明显差异,脑畸形血管患者血管组织可表达更多的血管内皮生长因子和转化生长因子α。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程全文链接：     

CXCR-4 AND CCR-5 expression in normal term human placenta

The maternal-fetal interphase has an active Immunitary System (IS) whose mediators -cells, cytokines and chemokines- coordinately act to favour pregnancy normal development. It is not known exactly which of those mediators are present in each placental cellular stratus and what the physiological or potentially pathologic consequences derived from their presence can be. It is known that chemokines recruit cells with regulatory activity towards the deciduous and some of their receptors are coreceptors to infectious agents like HIV, making research of chemokines expression and their receptors in the maternal-fetal interphase of great interest in recent years. In the present study, the CXCR-4 and CCR-5 expression was investigated in 8 samples of normal human placenta obtained from term pregnancies, with low obstetric risk, by using Immunocitochemical techniques (Biotin-Avidin-Peroxidase). The most relevant finding in this study was the demonstration that CXCR-4 and CCR-5 differential expression in trophoblast, stroma and endothelium represents, as far as we know, the first report of the presence of these receptors in all layers of placental tissue. These results help to broaden the knowledge about the expression of chemokines receptors -that act as main coreceptors in the HIV infection- in the maternal-fetal interphase, and this can be a contribution to be taken into account in the vertical transmission study of this infectious agent.     

Immunohistochemical expression of two members of the GDNF family of growth factors and their receptors in the olfactory system

The glial cell line-derived (GDNF) family of trophic factors, GDNF, neurturin, persephin and artemin, are known to support the survival and regulate differentiation of many neuronal populations, including peripheral autonomic, enteric and sensory neurons. Members of this family of related ligands bind to specific GDNF family receptor (GFR) proteins, which complex and signal through the Ret receptor tyrosine kinase. We showed previously that GDNF protein was detectable in olfactory sensory neurons (OSNs) in the olfactory neuroepithelium (ON). In this immunohistochemical study, we localized GDNF, neurturin, GFRα1, GFRα2 and Ret in the adult rat ON and olfactory bulb. We found that GDNF and Ret were widely expressed by immature and mature OSNs, while neurturin was selectively expressed in a subpopulation of OSNs zonally restricted in the ON. The GFRs had differential expression, with mature OSNs and their axons preferentially expressing GFRα1, whereas progenitors and immature neurons more avidly expressed GFRα2. In the bulb, GDNF was highly expressed by the mitral and tufted cells, and by periglomerular cells, and its distribution generally resembled that of Ret, with the exception that Ret was far more predominant on fibers than cell bodies. Neurturin, in contrast, was present at lower levels and was more restricted in its expression to the axonal compartment. GFRα2 appeared to be the dominant accessory protein in the bulb. These data are supportive of two members of this neurotrophic family, GDNF and neurturin, playing different physiological roles in the olfactory neuronal system.     

Expression of vascular endothelial growth factor receptors 1, 2 and 3 in placentas from normal and complicated pregnancies

Extensive angiogenesis and invasion of the maternal decidua by trophoblasts are essential for the development and function of the placenta. Vascular endothelial growth factors (VEGF), placenta growth factor (PlGF) and their receptors VEGFR-1/Flt-1, VEGFR-2/KDR and VEGFR-3/Flt4 have important roles in vasculogenesis and angiogenesis. We have studied the localization of these proteins by immunohistochemistry and Western blotting in the placenta and of PlGF in maternal serum, and their association with diabetes, pre-eclampsia, fetal growth restriction (FGR) and fetal alcohol syndrome (FAS). VEGFR-1 and VEGFR-3 were detected mainly in the syncytiotrophoblastic layer whereas VEGFR-2 was detected in the vascular endothelial cells of the placenta. VEGFR-1, but not the other receptors, showed increased expression in placental syncytiotrophoblasts from 50% of patients with severe pre-eclampsia and FGR when compared with normal placentas. PlGF was undetectable in 38 of 44 samples of amniotic fluid of mothers with normal and complicated pregnancies. However, maternal serum PlGF concentrations were significantly lower in pre-eclamptic patients and in those with FGR when compared to diabetic women or healthy controls. These results suggest that low maternal serum PlGF and increased placental expression of its receptor VEGFR-1 are associated with pre-eclampsia and FGR.     

Differential expression of type I insulin-like growth factor receptors in different stages of human T cells

Insulin-like growth factor I (IGF-I) has been implicated to play a regulatory role in T cell development and in T cell function. We investigated the expression of type I IGF receptors on human peripheral T cells related to the maturation and activation stage using the type I IGF receptor-specific monoclonal antibody αIR3. It appeared that 87% of the CD4⁺CD45RA⁺ cells and 66% of the CD8⁺CD45RA⁺ cells were αIR3⁺, whereas only 37% of the CD4⁺CD45RO⁺ cells and 38% of the CD8⁺CD45RO⁺ cells bound αIR3. We also found that the fraction of αIR3⁺ cells within in vivo or in vitro activated (HLA-DR⁺) T cells is markedly lower than in nonactivated (HLA-DR^?) cells. In vitro phytohemagglutinin-activated T cells and CD4⁺CD45RO⁺ cells activated with recall antigens also contained less αIR3⁺ cells (1–6%) than nonactivated cells (30–54%).     

Fc receptors in human placenta.

Cryostat sections of placental tissue strongly adsorbed erythrocytes sensitized with IgG antibodies of human, rabbit, and guinea pig origin. No adsorption occurred using erythrocytes sensitized with F(ab')2 fragments. The reaction was strongly inhibited by intact IgG and by Fc fragments, weakly inhibited by pFc' fragments, and not inhibited by Facb and F(ab')2 or albumin. These properties are similar to those of corresponding receptors in normal lymphoid tissues. Results obtained with sections of hydatidiform mole showed that the reaction occurred with the trophoblastic tissue. Porcine placenta had no Fc receptor activity. The presence of an Fc receptor in human placental tissue may therefore be of significance for the selective transfer of IgG from mother to foetus.     

Differential expression of oestrogen receptors in human secondary lymphoid tissues

Many autoimmune diseases including rheumatoid arthritis (RA), Sj?gren's syndrome (SS) and systemic lupus erythematosus (SLE) occur much more frequently in women than in men. There is much evidence that oestrogen is the major cause of this gender difference. Interestingly, oestrogen relieves the symptoms of RA and SS but it exacerbates SLE. This contradictory effect of oestrogen on autoimmune diseases is not well understood. Most of the effects of oestrogen are mediated by two receptors: oestrogen receptor alpha and beta (ERalpha and ERbeta). To determine whether these contradictory effects of oestrogen relate to the involvement of distinct effects of the two ERs, we investigated expression of ERalpha and ERbeta in human secondary lymphoid tissues. We observed that, in tonsils, ERbeta is expressed in lymphocytes of germinal centres (GC) and the follicular mantle zone as well as in granulocytes, while ERalpha is expressed only in activated germinal centres but not in the follicular zone. ERbeta is the predominant ER in human leucocytes from peripheral blood, spleen and in leucocytes infiltrating cancers in both males and females. In addition, in different human lymphoma cell lines including Hodgkin lymphoma, Burkitt lymphoma, and multiple myeloma, ERbeta is abundant while ERalpha is not detectable. Our results indicate that ERbeta is the predominant type of ER in mature lymphocytes. We suggest that ERalpha and ERbeta have distinct roles in secondary lymphoid tissues and that further studies with ERbeta-specific agonists will help to elucidate the role of ERbeta in these tissues.