Fine Structure and Ca-ATPase Activity of the Stratum Intermedium Cells During Odontogenesis in Gars,Lepisosteus , Actinopterygii

This is the first report on the stratum intermedium in vertebrates other than mammals. The aim of this study is to elucidate the fine structure and cytochemical features of the stratum intermedium during the stages of enameloid formation in Lepisosteus . Inner dental epithelium, stratum intermedium, stellate reticulum, and outer dental epithelium are consistently present in the tooth germs of Lepisosteus . The stratum intermedium cells are oval in shape, contain elliptical nuclei, and extend many small processes. It is implied that the structure of the enamel organ is different among actinopterygians, and that constitution of the enamel organ in Lepisosteus resembles that in higher vertebrates. Marked Ca-ATPase activity is observed at the cell membrane of the stratum intermedium cells, suggesting that the cells are involved in calcium transport during the stages of enameloid formation.     

Immunohistochemical localisation of the 25 kDa heat shock protein in unstressed rats: Possible functional implications

The distribution of the 25 kDa heat shock protein (hsp 25) in a number of tissue types from unstressed rats was investigated. Immunohistochemical analysis showed that hsp 25 was not found in the thymus, brain (cerebral cortex and cerebellum), testis, adrenal, liver, spleen, or kidney. A number of cells in the anterior pituitary showed strong staining. These cells were tentatively identified as being either gonadotropes or thyrotropes. Strong staining was also observed in the blood vessels within these tissues. Hsp 25 was found to be localised predominantly to intestinal smooth muscle of the duodenum and colon and to vascular smooth muscle. Smooth muscle from other sites, such as the trachea, was also intensely stained. Lower and more variable amounts of staining were observed in cardiac and skeletal muscle. These observations suggest that hsp 25 is associated with cytoskeletal elements in muscle, and that the high staining intensity in smooth muscle might be due to the lack of internal architecture present in this muscle type. © 1993 Wiley-Liss, Inc.     

Immunohistochemical Localization of SNARE Proteins in Dental Pulp and Periodontal Ligament of the Rat Incisor

Distribution of three soluble N‐ethylmaleimide‐sensitive fusion protein attachment protein receptor (SNARE) proteins, syntaxin‐1, synaptosomal‐associated protein of 25 kDa (SNAP‐25), and vesicle‐associated membrane protein‐2 (VAMP‐2), was examined in dental pulp and periodontal ligament of the rat incisor. In the trigeminal ganglion, syntaxin‐1 and SNAP‐25 immunoreactivity was predominately detected in medium‐ to large‐sized neurons. Most syntaxin‐1 immunoreactive neurons expressed SNAP‐25. In contrast, VAMP‐2 was localized in small‐ to medium‐sized neurons and in slender‐shaped cells surrounding SNAP‐25‐immunopositive neurons. When the inferior alveolar nerve, one of the mandibular nerve branches innervating the dental pulp and periodontal ligament, was ligated, SNARE proteins accumulated at the site proximal to the ligation. In the incisor dental pulp, all nerve fibers displayed immunoreactivity for syntaxin‐1, SNAP‐25, and VAMP‐2. In the periodontal ligament of the incisor, almost all nerve fibers displayed both syntaxin‐1 and SNAP‐25 immunoreactivity, but lacked VAMP‐2 immunoreactivity. SNAP‐25 protein expression was localized around the vesicle membranes at the axon terminal of the periodontal mechanoreceptors. These present data suggest that these three SNARE proteins are synthesized at the trigeminal ganglion, transported centrally and peripherally, and expressed in sensory endings where apparent synapses are not present. Because those proteins participate in docking and exocytosis of synapse vesicles in the central nervous system, they might also contribute to vesicle exocytosis at receptive fields where apparent synapses are not present. Anat Rec, 2010. © 2010 Wiley‐Liss, Inc.     

NGF对局灶性脑缺血再灌注大鼠海马CREB表达的上调作用

目的探讨外源性神经生长因子(NGF)对局灶性脑缺血再灌注大鼠海马CREB和磷酸化的CREB(p-CREB)表达的影响。方法用线栓法阻塞大鼠大脑中动脉制作局灶性脑缺血再灌注模型,应用免疫组织化学、W estern B loting和图像分析方法检测大鼠海马CA1区CREB和磷酸化CREB表达。结果假手术组海马CA1区CREB有明显表达,缺血再灌注组CA1区表达较假手术组减少,NGF组CA1区的CREB表达多于缺血再灌注组(P<0.05)。假手术组海马CA1区p-CREB表达很少,缺血再灌注组p-CREB表达多于假手术组,NGF组p-CREB表达多于缺血再灌注组(P<0.05)。结论NGF上调海马CREB和p-CREB的表达,对缺血神经元起保护作用,CREB和p-CREB参与NGF对缺血神经元的保护作用机制。     

三叶因子1在实验性胃溃疡自愈期大鼠下颌下腺的表达及意义

目的 探讨大鼠实验性胃溃疡自愈期间下颌下腺三叶因子1(TFF1)的表达变化.方法 采用免疫组织化学和RT-PCR法,分别从蛋白水平及基因水平检测42只溃疡组、21只盐水组和6只正常组大鼠下颌下腺TFF1的表达.结果 免疫组织化学SP法显示,正常大鼠下颌下腺颗粒曲管的多颗粒细胞呈TFF1免疫反应阳性,各级导管管腔内也有TFF1阳性物质,纹状管管腔游离面可见线条状TFF1阳性物.溃疡组下颌下腺TFF1肽积分吸光度值明显增加,高于相应的盐水对照组和正常组(P<0.05或P<0.01),其中1d、2d、4d、6d TFF1肽积分吸光度值逐渐增加,6d最高,10d、14d、23d也显著高于盐水对照组.RT-PCR结果 显示,下颌下腺有TFF1 mRNA转录,且溃疡组TFF1/GAPDH mRNA吸光度比值在溃疡2～23d均高于盐水对照组和正常组(P<0.01或P<0.05).结论 下颌下腺TFF1肽在大鼠实验性胃溃疡形成和愈合过程中高表达,主要通过导管系统排泄,参与实验性胃溃疡的愈合.     

Effects of benzo(a)pyrene on the expression of heat shock proteins, pro-inflammatory cytokines and antioxidant enzymes in hepatic tumors induced by rat hepatoma N1-S1 cells

Benzo(a)pyrene (BaP) is a polycyclic aromatic hydrocarbon (PAH) that is easily introduced to humans via consumption of grilled or smoked meat. BaP causes harmful oxidative effects on cell development, growth and survival through an increase in membrane lipid peroxidation, oxidative DNA damage and mutagenesis. Therefore, the present study was conducted to evaluate the synergistic effects of BaP on oxidative stress in hepatic tumors. In this study, we established a hepatic tumor model by injecting rat hepatoma N1-S1 cells into healthy rats. Changes in the abundance of heat shock proteins (HSPs), antioxidant enzymes and pro-inflammatory cytokines were then investigated by western blot analysis. In addition, we examined changes in oxidative stress levels. Injection of N1-S1 cells or concomitant injection of BaP and N1-S1 cells resulted in the formation of hepatic tumors at the injection site. Evaluation of rat plasma reveals that hepatic tumors induced by BaP and N1-S1 cells expresses higher levels of Hsp27, superoxide dismutase (SOD), and tumor necrosis factor-α (TNF-α) when compared to those induced by N1-S1 cells only. The collective results of this study suggest that BaP exerts synergistic effects on the expression of HSP, cytokines and antioxidant enzymes in hepatic tumors induced by rat hepatoma N1-S1 cells.     

Expression and Role of the BDNF Receptor-TrkB in Rat Adrenal Gland under Acute Immobilization Stress

We reported that plasma brain-derived neurotrophic factor (BDNF) was maximally elevated following a 60-min period of acute immobilization stress and that salivary glands were the main source of plasma BDNF under this stress condition. However, the expression pattern of the BDNF receptor, Tyrosine receptor kinase B (TrkB), under this condition has yet to be determined. We therefore investigated the effect of this stress on the expression level of TrkB in various rat organs using real-time PCR. No significant differences were found between controls and 60 min-stressed rats with respect to TrkB level in various organs. Only adrenal glands showed significantly increased TrkB mRNA levels after 60 min of stress. TrkB mRNA and protein were observed to localize in chromaffin cells. In addition, we investigated whether BDNF-TrkB interaction influences the release of stress hormones from PC12 cells, derived from chromaffin cells. Truncated receptor, TrkB-T1, was identified in PC12 cells using RT-PCR. Exposure of PC12 cells to BDNF induced the release of catecholamine. This BDNF-evoked release was totally blocked by administration of the K252a in which an inhibitor of Trk receptors. Thus, BDNF-TrkB interactions may modulate catecholamine release from adrenal chromaffin cells under acute stress conditions.     

1alpha,25-dihydroxyvitamin D3 increases IgA serum antibody responses and IgA antibody-secreting cell numbers in the Peyer's patches of pigs after intramuscular immunization

Pigs were injected intramuscularly (i.m.) twice with human serum albumin (HSA) with or without 1alpha,25-dihydroxyvitamin D3[1alpha,25(OH)2D3] with a 5-week interval. The supplementation of 1alpha,25(OH)2D3 enhanced the HSA-specific IgA serum antibody response but decreased the IgM, IgG, IgG1 and IgG2 responses. Furthermore, higher numbers of HSA-specific IgA antibody-secreting cells were obtained in systemic lymphoid tissues (local draining lymph node, spleen and bone marrow) as well as in Peyer's patches and lamina propria of the gut (GALT). In addition, the in vivo mRNA expression for Th1 [interferon (IFN)-gamma, interleukin (IL-2)], Th2 (IL-4, IL-6 and IL-10) and Th3 [transforming growth factor (TGF)-beta] cytokines as well as the percentage of different cell subsets (CD2+, CD4+, CD8+, IgM+, MHC II+, CD25+) of monomorphonuclear cells from the local draining lymph node were determined at different time-points after the i.m. immunizations. Cytokine profiles did not resemble a typical Th-cytokine profile using 1alpha,25(OH)2D3: higher levels of IL-10 and significantly lower levels of IL-2 were observed the first day after the primary immunization. However, significantly higher levels of IL-2 and significantly lower levels of IFN-gamma were observed the first day after the second immunization. Furthermore, after the second immunization TGF-beta mRNA expression decreased more quickly in the 1alpha,25(OH)2D3 group. This difference became significant 7 days after the second immunization. One week later a significantly higher percentage of CD25+ cells was observed in this group, indicating more activated T and B cells using the steroid hormone. These results suggest that in pigs the addition of 1alpha,25(OH)2D3 to an intramuscularly injected antigen can enhance the antigen-specific IgA-response and prime GALT tissues, but the relation with cytokines and cell phenotype in the local draining lymph node needs further clarification.     

Expression of CD25 is a specific and relatively sensitive marker for the Philadelphia chromosome (BCR-ABL1) translocation in pediatric B acute lymphoblastic leukemia

Background: Precursor B acute lymphoblastic leukemia (B-ALL) is the most common cancer in children and overall, has an excellent prognosis. However, the Philadelphia chromosome translocation (Ph+), t(9;22)(q34;q11), is present in a small subset of patients and confers poor outcomes. CD25 (IL-2 receptor alpha chain) expression has been associated with Ph+ B-ALL in adults, but no similar study has been performed in pediatric B-ALL. Methods: A retrospective analysis of 221 consecutive pediatric patients with a diagnosis of B-ALL (blood and/or bone marrow) from 2009 to 2012 was performed to determine an association between Ph+ B-ALL and CD25 expression. A threshold of 25% was used to define positive cases for CD25 expression by flow cytometry. Results: There were 221 patients with a diagnosis of B-ALL ranging from 2 to 22 years (median, 6 years). Eight (3.6%) B-ALL patients were positive for the Philadelphia chromosome translocation (Ph+ B-ALL) and 213 were negative (Ph-negative B-ALL). CD25 expression was observed in 6 of 8 (75%) Ph+ B-ALL patients and 6 of 213 (2.8%) Ph-negative B-ALL patients. CD25 expression was significantly higher in Ph+ B-ALL compared to Ph-negative B-ALL, with median CD25 expression of 64% (range 0-93%) and 0.1% (range 0-91%), respectively (P ≤ 0.0002). Therefore, CD25 expression as a predictor of Ph+ B-ALL had 75% sensitivity, 97% specificity, 50% positive predictive value and 99% negative predictive value. Conclusions: CD25 expression is a specific and relatively sensitive marker for the identification of Ph+ B-ALL in the pediatric population.     

NGF和CGRP对局灶性脑缺血再灌注后大鼠皮质CHOP蛋白表达的影响

目的探讨外源性神经生长因子(NGF)和降钙素基因相关肽(CGRP)对局灶性脑缺血再灌注后大鼠顶叶皮质神经元CHOP蛋白表达的影响及其作用机制。方法用线栓法制备大鼠大脑中动脉阻塞(MCAO)模型,应用免疫组织化学方法,免疫印迹法(W estern B lotting)测定CHOP蛋白表达;M etamoph图像分析系统对结果进行分析。结果假手术组大鼠顶叶皮质未见CHOP蛋白表达;缺血组CHOP蛋白在缺血再灌注后12 h明显表达,24 h达高峰,72 h显著下降,缺血组较假手术组CHOP蛋白表达显著增加(P<0.01);NGF组和CGRP组CHOP蛋白表达分别弱于缺血组(P<0.01);NGF和CGRP合用组CHOP蛋白表达明显弱于缺血组(P<0.01),也分别弱于NGF组和CGRP组(P<0.01)。结论NGF和CGRP能分别下调局灶性脑缺血再灌注大鼠顶叶皮质CHOP蛋白表达,联合应用则作用更强,二者对保护脑缺血神经元可能有协同作用。