趋化因子CXCL12及其受体CXCR4与造釉型颅咽管瘤侵袭、复发的相关性研究

目的探讨趋化因子及其受体(CXCL12/CXCR4)在造釉细胞型颅咽管瘤中的表达并分析其与肿瘤侵袭、复发的相关性。方法应用免疫组化法和western blot方法对20例造釉细胞型颅咽管瘤标本(原发组11例,复发组9例)中CXCL12/CXCR4的表达进行检测。结果 CXCL12/CXCR4在复发组造釉细胞型颅咽管瘤中的阳性表达明显高于原发组(P<0.05)。结论 CXCL12/CXCR4可能在造釉细胞型颅咽管瘤侵袭、复发过程中起重要的作用。CXCL12/CXCR4可能成为评价造釉细胞型颅咽管瘤侵袭性生长和复发的标记物。     

炎症相关因子IFN-α及TGF-β1在颅咽管瘤的表达及意义

目的探讨炎症相关因子IFN-α、TGF-β1与颅咽管瘤炎症反应的关系。方法采用免疫组化GTVisionTM法检测α-干扰素(interferon-α,IFN-α)、转化生长因子-β1(transforming growth factorβ1,TGF-β1)在58例颅咽管瘤组织中的表达情况,比较不同病理类型颅咽管瘤、复发组及初发组颅咽管瘤组织中IFN-α、TGF-β1的表达水平。结果造釉细胞型及复发颅咽管瘤IFN-α表达广泛。44例造釉细胞型颅咽管瘤有40例IFN-α表达阳性(90.91%),鳞状乳头型14例中有12例表达阳性,造釉细胞型IFN-α阳性表达率明显高于鳞状乳头型(Z=-2.003,P<0.05)。16例复发颅咽管瘤均有IFN-α阳性表达,阳性表达率高于初发组(Z=-2.085,P<0.05)。TGF-β1在鳞状乳头型颅咽管瘤明显表达,14例中有10例阳性表达,而造釉细胞型44例仅有14例表达阳性(Z=-2.129,P<0.05)。经Spearman相关分析,在全部颅咽管瘤中,IFN-α与TGF-β1表达程度呈负相关(rs=-0.273,P<0.05)。结论颅咽管瘤存在复杂的炎症反应,炎症相关因子IFN-α、TGF-β1可能参与颅咽管瘤细胞的生长调控,两者之间可能存在拮抗作用。     

CD147、MMP-9与釉质型颅咽管瘤侵袭性生长及复发的相关性研究

目的:探讨细胞外基质金属蛋白酶诱导剂( CD147,EMMPRIN)和基质金属蛋白酶-9(MMP-9)在釉质型颅咽管瘤中的表达及其与肿瘤侵袭、复发的相关性。方法:应用免疫组化法对40例釉质型颅咽管瘤标本（原发组24例,复发组16例）中CD147和MMP-9的表达进行检测。结果:CD147在复发组釉质型颅咽管瘤中的总阳性表达率87.5％,明显高于原发组总阳性率37.5％ (P＜0.05);MMP-9在复发组釉质型颅咽管瘤中的总阳性表达率93.7％,明显高于原发组总阳性率41.7％ (P ＜0.05)。复发组釉质型颅咽管瘤中CD147与MMP-9表达呈正相关（P＜0.01）,原发组釉质型颅咽管瘤中CD147与MMP-9表达无明显相关性。结论: CD147与MMP-9可能在釉质型颅咽管瘤侵袭生长和肿瘤复发过程之中扮演着重要的角色,CD147与MMP-9可能成为评价釉质型颅咽管瘤侵袭性生长和复发倾向的重要参考指标。     

MVD、VEGF及Ki-67在颅咽管瘤中的表达与意义

目的探讨颅咽管瘤中MVD、VEGF及Ki-67表达与意义。方法根据病理亚型、形态学特征及复发情况,将47例颅咽管瘤按不同分类方法分为成釉质细胞型30例,鳞状乳头型17例;原发31例,复发16例;实性为主肿瘤10例,囊性为主肿瘤37例。SP法检测肿瘤标本MVD、VEGF及Ki-67表达,统计分析不同分类的组间表达差异。结果鳞状乳头型肿瘤MVD、VEGF的表达明显高于成釉质细胞型(均P0.001),而Ki-67在两种病理亚型之间无显著差异(P0.05);囊性为主的肿瘤中VEGF表达明显高于实性为主的肿瘤(P=0.018),而MVD、Ki-67在两种形态学分组之间无显著差异(均P0.05);原、复发组间MVD、VEGF及Ki-67均无显著差异(均P0.05)。在本组颅咽管瘤中,MVD与VEGF显著相关(P=0.014),两者均与Ki-67无显著相关(均P0.05)。结论 MVD、VEGF的测定可为颅咽管瘤的血管发生提供依据,VEGF的表达与肿瘤囊性结构形成密切相关,单独检测MVD、VEGF和Ki-67其中一项指标不能预测颅咽管瘤复发。     

颅咽管瘤中BCL-2蛋白的表达及意义

目的　分析颅咽管瘤中凋亡抑制基因BCL - 2在蛋白水平的表达 ,探讨其在颅咽管瘤生长机制中的意义。方法　选取经手术及病理证实的颅咽管瘤 4 9例 ,采用免疫组化法检测BCL 2蛋白的表达。结果　BCL 2的阳性表达程度在不同病理类型的颅咽管瘤上有显著差异 ,造釉细胞型颅咽管瘤的表达强度显著高于鳞状乳头型颅咽管瘤 (P <0 0 1)。结论　BCL 2在不同病理类型的颅咽管瘤中的表达呈现明显的区别 ,提示 :凋亡调控机制的失控可能是造釉型颅咽管瘤生长的重要机制之一 ;胚胎残余组织的继续增殖学说更能解释造釉细胞型颅咽管瘤的起源可能。     

颅咽管瘤中基质金属蛋白酶MMP-2的表达及意义

目的探讨颅咽管瘤局部侵袭性生长的生物学机制。方法使用免疫组织化学S-P法检测基质金属蛋白酶MMP-2在14例颅咽管瘤细胞中的表达。结果2例乳头型、9例造釉细胞型及1例混合型颅咽管瘤均有不同程度的表达。MMP-2阳性染色主要位于肿瘤细胞的胞质中,尤其以鳞状上皮的棘细胞和造釉上皮的近基底层细胞的表达最为明显。1例囊状复发及1例γ-刀术后造釉细胞瘤型未见表达。结论3种类型颅咽管瘤细胞均能产生MMP-2,提示MMP-2可能是颅咽管瘤局部侵袭的机制之一。     

颅咽管瘤鸡胚绒毛尿囊膜移植瘤模型的建立

目的初步建立颅咽管瘤鸡胚绒毛尿囊膜(chorioallantoic membrane CAM)移植瘤模型,并评估移植瘤血管生成能力及细胞增殖能力。方法利用术中所得颅咽管瘤组织,异种移植到鸡胚CAM表面,建立移植瘤模型。对移植瘤瘤体周围CAM血管进行测定,评估移植瘤CAM血管生成能力,同时利用免疫组化法以CD34标记移植瘤微血管密度(microvessel density,MVD),以PCNA检测移植瘤细胞增殖活性。结果成功建立颅咽管瘤CAM移植瘤模型,造釉细胞型颅咽管瘤成瘤率为47.14%,鳞状上皮型颅咽管瘤成瘤率为43.33%,两者无显著差异(χ2=0.123,P=0.726);造釉细胞型移植瘤瘤周CAM血管生成能力,微血管密度及细胞增殖活性均高于鳞状上皮型移植瘤。移植瘤PCNA的表达与MVD(Pearson r=0.639,P0.001)及CAM血管评分(Spearman r=0.490,P=0.001)呈正相关。结论采用异位异种移植的方法可在鸡胚CAM上建立颅咽管瘤移植瘤模型。移植瘤可不同程度地刺激CAM上血管生成,移植瘤自身可建立微循环,具有一定的增殖能力。     

颅咽管瘤β-连接素表达及其基因突变分析

目的分析不同病理类型颅咽管瘤的β-连接素表达特征及基因外显子3突变的意义。方法研究我院自2004年1月至2006年1月共23例颅咽管瘤标本,其中造釉细胞型颅咽管瘤20例、鳞状乳头型3例,应用免疫组化方法检测β-连接素表达特征,同时分析其基因外显子3的突变情况。结果免疫组化结果显示,β-连接素在造釉细胞瘤型颅咽管瘤的细胞核及细胞浆中表达,而在鳞状乳头型颅咽管瘤中仅见细胞膜表达;基因突变分析显示,40%的造釉细胞型颅咽管瘤β-连接素基因在外显子3上发生突变,鳞状乳头型无此突变。结论造釉细胞型颅咽管瘤和鳞状乳头型颅咽管瘤不仅在临床病理上,而且在分子水平也存在差异。β-连接素基因外显子3的突变在造釉细胞型颅咽管瘤的发生、发展中可能起一定作用。     

不同类型颅咽管瘤的组织炎症和细胞增殖性

目的研究不同病理类型颅咽管瘤的组织炎症及细胞增殖性对预后的影响.方法对30例经手术切除的颅咽管瘤组织采用免疫组织化学SP法检测白细胞共同抗原CD45和细胞周期相关核抗原Ki-67的抗体MIB-1的表达,分别计算CD45标记指数(CD45 LI)和MIB-1标记指数(MIB-1 LI).结果CD45标记的炎症反应在颅咽管瘤组织中广泛存在,MIB-1主要在颅咽管瘤基底细胞层表达.造釉细胞型颅咽管瘤组织CD45 LI(32.4%±15.0%)及细胞MIB-1 LI(18.2%±8.7%)显著性高于鳞状乳头型颅咽管瘤(分别为13.6±7.6%、7.1%±4.9%,P＜0.05).结论颅咽管瘤造釉细胞性较鳞状乳头型具有更强的组织炎症和细胞增殖性,可能是影响其预后的原因.     

SYBR荧光实时定量PCR检测不同病理类型颅咽管瘤ICAM-1 mRNA表达

目的定量研究细胞间粘附分子基因(ICAM—1 mRNA)在不同病理类型颅咽管瘤的表达差异及意义。方法收集30例经手术治疗的颅咽管瘤标本,采用SYBR荧光实时定量PCR法检测 ICAM-1 mRNA在肿瘤组织的表达,并对表达结果行统计学分析。结果造釉细胞型颅咽管瘤 ICAM-1mRNA表达量为(62．18±6．43)×103 copies／μg,鳞状乳头型颅咽管瘤ICAM-1 mRNA表达量为 (1．13±0．17)×103 copies／μg,造釉细胞型颅咽管瘤ICAM-1 mRNA表达量显著性高于鳞状乳头型颅咽管瘤(P<0．01)。结论两种病理类型颅咽管瘤ICAM—1 mRNA表达存在显著性差异,此差异性可能与两种病理类型颅咽管瘤不同的肿瘤炎症有关。     

Development of the projection from the nucleus of the brachium of the inferior colliculus to the superior colliculus in the ferret

Neurons in the deeper layers of the superior colliculus (SC) have spatially tuned receptive fields that are arranged to form a map of auditory space. The spatial tuning of these neurons emerges gradually in an experience-dependent manner after the onset of hearing, but the relative contributions of peripheral and central factors in this process of maturation are unknown. We have studied the postnatal development of the projection to the ferret SC from the nucleus of the brachium of the inferior colliculus (nBIC), its main source of auditory input, to determine whether the emergence of auditory map topography can be attributed to anatomical rewiring of this projection. The pattern of retrograde labeling produced by injections of fluorescent microspheres in the SC on postnatal day (P) 0 and just after the age of hearing onset (P29), showed that the nBIC-SC projection is topographically organized in the rostrocaudal axis, along which sound azimuth is represented, from birth. Injections of biotinylated dextran amine-fluorescein into the nBIC at different ages (P30, 60, and 90) labeled axons with numerous terminals and en passant boutons throughout the deeper layers of the SC. This labeling covered the entire mediolateral extent of the SC, but, in keeping with the pattern of retrograde labeling following microsphere injections in the SC, was more restricted rostrocaudally. No systematic changes were observed with age. The stability of the nBIC-SC projection over this period suggests that developmental changes in auditory spatial tuning involve other processes, rather than a gross refinement of the projection from the nBIC.     

The organization of the connections between the cortex and the claustrum in the monkey

The distribution of labelled cells and of extracellular granules in the claustrum has been studied after injections of horseradish peroxidase in several areas of the neocortex. The frontal and parietal lobes are related to the anterior and posterior halves respectively of the claustrum, and the occipital and temporal cortex to the posterior and inferior margins. Parts of the claustrum related to areas of the cortex in the frontal lobe overlap considerably in the antero-posterior dimension with parts related to widely separated but interconnected areas of the parieto-temporal cortex. Areas of cortex within one lobe which are interconnected are related to parts of the claustrum which overlap in the dorsoventral dimension.     

Sexual pheromones and the evolution of the reward system of the brain: the chemosensory function of the amygdala

The amygdala of all tetrapod vertebrates receives direct projections from the main and accessory olfactory bulbs, and the strong similarities in the organization of these projections suggest that they have undergone a very conservative evolution. However, current ideas about the function of the amygdala do not pay sufficient attention to its chemosensory role, but only view it as the core of the emotional brain. In this study, we propose that both roles of the amygdala are intimately linked since the amygdala is actually involved in mediating emotional responses to chemical signals. The amygdala is the only structure in the brain receiving pheromonal information directly from the accessory olfactory bulbs and we have shown in mice that males emit sexual pheromones that are innately attractive for females. In fact, sexual pheromones can be used as unconditioned stimuli to induce a conditioned attraction to previously neutral odorants as well as a conditioned place preference. Therefore, sexual pheromones should be regarded as natural reinforcers. Behavioural and pharmacological studies (reviewed here) have shown that the females' innate preference for sexual pheromones is not affected by lesions of the dopaminergic cells of the ventral tegmental area, and that the systemic administration of dopamine antagonists do not alter neither the attraction nor the reinforcing effects of these pheromones. Anatomical studies have shown that the vomeronasal amygdala gives rise to important projections to the olfactory tubercle and the islands of Calleja, suggesting that these amygdalo-striatal pathways might be involved in the reinforcing value of sexual pheromones.     

Evidence for the collateral innervation of the esophagus and the heart from neurons in the compact formation of the nucleus ambiguus of the rat

We investigated whether the heart receives collateral projections from the neurons which innervate the esophagus with a retrograde double-labeling method using two fluorescent tracers. Following injections of True Blue (TB) into the esophagus and Diamidino Yellow (DY) into the heart, about 21.9% of the labeled esophageal motoneurons in the compact formation of the nucleus ambiguus (AmC) were retrogradely double labeled. No single-labeled cardiac motoneurons were found in the AmC. The present results provide anatomical substrates for the esophagocardiac reflex.     

A projection to the striatum from the medial subdivision of the posterior group of the thalamus in the cat

Injections of wheat germ agglutinin-conjugated horseradish peroxidase in the lateral part of the caudate nucleus or the putamen of the cat result in retrograde thalamic cell-labeling in the rostral extension of the medial subdivision of the posterior group (POM). Autoradiography after [³H]amino acid injection of POM reveals a dense and discontinuous distribution of axons in the lateral half of the caudate and putamen concentrated at their middle rostrocaudal levels. This newly discovered thalamostriatal projection of POM may account for somatosensory activity observed in striatal cells.     

Role of the paraventricular nucleus in the projection from the nucleus of the solitary tract to the olfactory bulb

Electrophysiological experiments were performed on anesthetized rats to determine the effects of lesions of the paraventricular nucleus on the amplitude of evoked potentials recorded in the periglomerular layer of the olfactory bulb after nucleus of the solitary tract electrical stimulation. Lesions of the paraventricular nucleus enhance the amplitude of both the positive and negative components of the evoked potential in the olfactory bulb. The pathway from the paraventricular nucleus to the olfactory bulb seems to exert a suppressive influence over the projection from the nucleus of the solitary tract to the olfactory bulb under these conditions.     

Projections from the five divisions of the orbital cortex to the thalamus in the rat

The orbital cortex (ORB) of the rat consists of five divisions: the medial (MO), ventral (VO), ventrolateral (VLO), lateral (LO), and dorsolateral (DLO) orbital cortices. No previous report has comprehensively examined and compared projections from each division of the ORB to the thalamus. Using the anterograde anatomical tracer, Phaseolus vulgaris leucoagglutinin, we describe the efferent projections from the five divisions of the ORB to the thalamus in the rat. We demonstrated that, with some overlap, each division of the ORB distributed in a distinct (and unique) manner to nuclei of the thalamus. Overall, ORB projected to a relatively restricted number of sites in the thalamus, and strikingly distributed entirely to structures of the medial/midline thalamus, while completely avoiding lateral regions or principal nuclei of the thalamus. The main termination sites in the thalamus were the paratenial nucleus (PT) and nucleus reuniens (RE) of the midline thalamus, the medial (MDm) and central (MDc) divisions of the mediodorsal nucleus, the intermediodorsal nucleus, the central lateral, paracentral, and central medial nuclei of the rostral intralaminar complex and the submedial nucleus (SM). With some exceptions, medial divisions of the ORB (MO, VO) mainly targeted “limbic-associated” nuclei such as PT, RE, and MDm, whereas lateral division (VLO, LO, DLO) primarily distributed to “sensorimotor-associated” nuclei including MDc, SM, and the rostral intralaminar complex. As discussed herein, the medial/midline thalamus may represent an important link (or bridge) between the orbital cortex and the hippocampus and between the ORB and medial prefrontal cortex. In summary, the present results demonstrate that each division of the orbital cortex projects in a distinct manner to nuclei of the thalamus which suggests unique functions for each division of the orbital cortex.     

On the development of the stratification of the inner plexiform layer in the chick retina

This study investigated the development of the subdivision of the chick inner plexiform layer (IPL). The approach included an immunohistological analysis of the temporal and spatial expressions of choline acetyltransferase, of the neural-glial-related and neural-glial cell adhesion molecules (NrCAM and NgCAM, respectively) and axonin-1, and of inwardly rectifying potassium (Kir) channels in 5- to 19-day-old (E5-E19) embryos. Ultrastructural investigations evaluated whether synaptogenesis accompanies the onset of differentiation of the IPL. We found that the differentiation of the IPL started at E9. Distinct cholinergic strata appeared, NrCAM immunoreactivity showed a poorly defined stratification, and Kir3.2 was expressed in the IPL and in the inner nuclear layer. From E10 until late E14, NgCAM- and axonin-1-immunoreactive strata emerged in an alternating sequence from the outer to the inner IPL. During this period, the NrCAM pattern sharpened, and eventually five bands of weaker and stronger immunoreactivity were found. Conventional synapses formed at the beginning of E9, and stratification of the IPL also began on the same day at the same location. Synaptogenesis and stratification followed a gradient from the central to the peripheral retina. The topographic course of differentiation of the IPL generally corresponded to the course of maturation of ganglion and amacrine cells. Synaptogenesis and the expression of G-protein-gated Kir3.2 channels accompanied the onset of stratification. These events coincide with the occurrence of robust and rhythmic spontaneous neuronal activity. The subsequent differentiation of the IPL seemed to be orchestrated by several mechanisms.     

Origins of the sympathetic innervation of the cervical end of the uterus in the rat

A retrograde neuronal tracer (Fast Blue) was injected in the cervical end of the uterine horn of virgin rats. The majority of the retrogradely labeled post-ganglionic sympathetic neurons were found in the sympathetic chain (74%). The superior mesenteric ganglia, inferior mesenteric ganglia and suprarenal ganglia accounted for 22, 3 and <1%, respectively. The distribution of neurons in the sympathetic chain labeled from the uterus resembles that described for other pelvic organs.