免疫荧光法观察P物质神经纤维在大鼠牙髓及牙周组织中的分布

目的:研究大鼠牙髓及牙周组织中神经肽SP免疫阳性神经纤维的分布情况。方法:采用冰冻切片和间接免疫荧光方法观察大鼠牙髓、牙周膜以及牙龈内SP免疫阳性神经纤维的表达。结果:SP免疫阳性神经纤维在大鼠牙龈和牙髓组织中分布较多,而在牙周组织中分布较少,并且主要分布在根中和根尖1/3的部位。无论在牙髓或牙周,SP免疫阳性神经纤维多沿血管分布。结论:牙髓及牙周组织中均有SP免疫阳性神经纤维的分布,并可能通过调节牙髓和牙周血管功能等途径,在其损伤和炎症反应中发挥作用。     

空气喷砂技术对牙髓组织中P物质作用的实验研究

目的：观察空气喷砂技术去龋时对牙髓组织的SP、CGRP神经反应性的影响。方法：实验组为空气喷砂技术组及高速涡轮机组,未制洞大鼠为空白对照组,制备雌性SD大鼠上颌磨牙,ABC免疫组织化学染色法观察牙髓组织内P物质的反应。结果：空气喷砂组大鼠牙髓SP、CGRP强阳性染色,高速涡轮机组大鼠牙髓SP、CGRP弱阳性染色,实验组间、实验组与对照组间均有差别,差别具有统计学意义（P〈0.05）。结论：空气喷砂技术刺激牙髓神经合成SP、CGRP增多,推论该技术在制备窝洞时对牙髓组织刺激性相对较小。     

大鼠腰椎P物质和降钙素基因相关肽能神经纤维的分布

目的　观察腰椎小关节 ,椎间盘 ,前、后纵韧带 ,棘上、棘间韧带上 P物质 (SP)和降钙素基因相关肽 (CGRP)能神经纤维的分布 ,探讨其意义。方法　采用 2 5例 SD大鼠腰椎组织切片 ,以SP、CGRP多克隆抗体进行免疫组织化学研究。结果　 SP、CGRP能神经纤维在腰椎上有广泛的分布。除滑膜皱襞外 ,小关节的其他部位均观察到上述神经纤维。在椎间盘 ,神经纤维只分布于纤维环的最外层。棘间韧带上仅见 CGRP能神经纤维。结论　腰椎上有广泛的感觉神经分布 ,上述结构参与慢性下腰痛发生有其神经解剖学基础     

背根神经节内P物质、降钙素基因相关肽免疫阳性神经元与阴茎包皮系带感觉信息的传递

目的：探讨背根神经节（DRG）内P物质（SP）、降钙素基因相关肽（CGRP）免疫阳性神经元与阴茎包皮系带感觉信息传递之问的关系。方法：通过荧光金（FG）逆行标记对大鼠阴茎包皮系带内神经末梢的来源作追踪定位，并结合SP、CGRP免疫荧光标记法，研究大鼠DRG内FG标记阳性神经元中SP、CGRP免疫阳性神经元的形态和分布。结果：FG逆行标记结果发现，大鼠阴茎包皮系带内的神经末梢起源于第6腰髓对应的背根神经节（L6-DRG）和第1骶髓对应的背根神经节（S1-DRG）的神经元。对这些神经元分别作SP、CGRP免疫荧光标记后发现，标记细胞大小不等，分别呈深红色和深绿色，沿神经束成行排列或散在分布。FG／SP、FG／CGRP双标记阳性细胞均为中小型，其数量分别占FG逆行标记阳性细胞总数的1／3和1／2，FG／SP／CGRP三标记阳性细胞占FG逆行标记阳性细胞总数的1／5。结论：大鼠L6-DRG和S1-DRG内的SP、CGRP免疫阳性神经元可能参与阴茎包皮系带感觉信息的传递。     

背根神经节内P物质、降钙素基因相关肽免疫阳性神经元与阴茎包皮系带感觉信息的传递

目的:探讨背根神经节(DRG)内P物质(SP)、降钙素基因相关肽(CGRP)免疫阳性神经元与阴茎包皮系带感觉信息传递之间的关系。方法:通过荧光金(FG)逆行标记对大鼠阴茎包皮系带内神经末梢的来源作追踪定位,并结合SP、CGRP免疫荧光标记法,研究大鼠DRG内FG标记阳性神经元中SP、CGRP免疫阳性神经元的形态和分布。结果:FG逆行标记结果发现,大鼠阴茎包皮系带内的神经末梢起源于第6腰髓对应的背根神经节(L6-DRG)和第1骶髓对应的背根神经节(S1-DRG)的神经元。对这些神经元分别作SP、CGRP免疫荧光标记后发现,标记细胞大小不等,分别呈深红色和深绿色,沿神经束成行排列或散在分布。FG/SP、FG/CGRP双标记阳性细胞均为中小型,其数量分别占FG逆行标记阳性细胞总数的1/3和1/2,FG/SP/CGRP三标记阳性细胞占FG逆行标记阳性细胞总数的1/5。结论:大鼠L6-DRG和S1-DRG内的SP、CGRP免疫阳性神经元可能参与阴茎包皮系带感觉信息的传递。     

VEGF、SP、CGRP及PCNA在门脉高压大鼠结肠黏膜中的表达

目的：探讨血管内皮生长因子、P物质、降钙素基因相关肽及增殖细胞核抗原在门脉高压大鼠黏膜中的表达及其意义。方法：20只SD大鼠随机分为两组，实验组大鼠行门静脉两步结扎加左肾上腺静脉结扎，对照组为假手组；应用免疫组化方法检测VEGF、SP、CGRP及PCNA在大鼠结肠黏膜中的表达情况。结果：实验组SP、CGRP及PCNA较对照组表达增强，VEGF无明显变化。结论：SP、CGRP可能参与门脉高压大鼠结肠黏膜局部病变。     

P物质阳性神经纤维在退变腰椎间盘的分布及意义

目的研究P物质（substance P,SP）阳性神经纤维在腰椎间盘中的出现及分布,以探讨其在椎间盘退变中的潜在作用。方法收集16例患者的21个病变腰椎间盘和来自于新鲜尸体的12个正常对照椎间盘,行组织学检查和SP免疫组织化学染色检查。结果SP阳性神经纤维偶见于正常椎间盘,在病变椎间盘内可见较多的SP免疫反应阳性神经纤维分布,阳性神经纤维数量与腰椎间盘退变程度呈正相关。结论腰椎间盘退变程度和SP阳性神经纤维数量有明显的相关性,SP可能作为神经炎性介质加速了腰椎间盘的退变。     

阴茎勃起神经递质CGRP蛋白表达的雄激素依赖性研究

目的 研究大鼠阴茎神经组织降钙素基因相关肽(CGRP)蛋白表达的雄激素依赖性。方法 64只雄性10周龄Sprague Dawley大鼠分为4组：A组为对照组，B组为去势组，C组为去势后肌注十一酸睾酮组，D组为喂饲保列治组。模型建立后分别在4周和10周处死部分大鼠，取腔静脉血和阴茎组织，应用放射免疫技术测定血睾酮(T)和二氢睾酮(DHT)浓度，应用免疫组化技术和计算机图象分析系统观察阴茎组织OGRP阳性神经纤维面积比例。结果 各实验组T和DHT含量低于对照组。4周时实验组和对照组阳性神经纤维面积比例的差异无统计学意义。10周时对照组阳性神经纤维面积比例明显高于实验组。各实验组4周模型和10周模型两两比较阳性神经纤维面积比例前者大于后者；对照组4周模型和10周模型阳性神经纤维面积比例无差异。结论 雄激素水平降低可引起大鼠阴茎神经组织CGRP蛋白表达下调，该影响有时间依从性。     

腰椎小关节退变致腰腿痛机制的实验研究

目的：探讨退变腰椎小关节病变和腰腿疼的关系及其可能的作用机制。方法：收集我院骨科临床手术中切除的腰椎小关节标本，利用组织病理学和影像学检查对比研究正常和退变腰椎小关节的病变特点，免疫组织化学方法观察P物质（substance P，SP）、降钙素基因相关肽（calcitonin gene-related peptide．CGRP）能神经纤维在小关节中的分布，酶联免疫吸附剂测定法（enzyme-linked immunosobnent assay，ELISA）测定腰椎小关节中细胞因子IL-1β的含量。结果：退变腰椎小关节的X线和CT表现为关节间隙有不同程度的变窄或消失、关节面骨质增生、硬化，HE染色软骨表面不平整、细胞排列紊乱；白介素-1β（interleukin-1β，IL-1β）水平升高。在两组小关节中都发现了SP、CGRP能神经纤维的存在。结论：腰椎小关节退变后发生了不同程度的病理变化，机械摩擦和细胞因子刺激小关节中的神经纤维可能是小关节病变引起腰腿疼的原因之一。     

烫伤大鼠肠道P物质含量和P物质能神经的观察分析

以30%TBSAⅢ度烫伤大鼠为模型,采用 RIA 和 IHC 的方法,动态观察了伤后72小时大鼠空肠免疫活性 P 物质(iSP)的含量及绒毛内 P 物质—免疫反应(SP—IR)阳性神经纤维的变化。结果发现:①烫伤大鼠空肠内 iSP 伤后1小时出现显著升高,4小时仍处于高水平状态;8小时后明显下降,伤后72小时一直处于低水平状态。③绒毛内 SP—IR 阳性神经纤维在形态、分布密度和纤维内 SP—IR 物质的含量都有明显的改变。图像分析定量结果表明:绒毛内 SP—IR 阳性神经纤维分布密度,伤后1小时明显增加,随后减少,48小时再次出现增加;神经纤维内 SP—IR 阳性物质伤后1小时及4小时明显升高,12小时明显下降,24小时后再次升高。实验结果说明:烫伤后大鼠空肠内 P 物质出现应激性释放与消耗,P 物质可能参与了烫伤后大鼠的肠道损伤;肠绒毛内的 P 物质能神经纤维是肠粘膜屏障损伤的直接参与因素。     

Immunohistochemical study of intralaryngeal ganglia in the cat.

To study the mechanism of autonomic regulation in the larynx, intralaryngeal local ganglia of the cat were investigated using immunohistochemical techniques. Small intralaryngeal ganglia were found in the peripheral portions of internal branches of the superior laryngeal nerve. Ninety-one percent of the ganglionic neurons were immunoreactive (IR) to vasoactive intestinal polypeptide (VIP), and 10% of the VIP-IR cells were also immunoreactive to enkephalin (ENK) and/or substance P (SP). The immunoreactivity of neuronal cell bodies remained unchanged even after denervation of the bilateral superior and recurrent laryngeal nerves. A dense distribution of calcitonin gene-related peptide (CGRP)-IR nerve fibers was found around almost all neuronal cells in the intralaryngeal ganglia. A few VIP-IR, ENK-IR, and SP-IR nerve fibers were also observed. Only the CGRP-IR fibers disappeared after the denervation experiments. In the laryngeal glands and mucosal arterioles, VIP-IR nerve terminals were found that were also immunoreactive to ENK and/or SP. However, these immunoreactive nerve endings in the glands and arterioles remained after the denervation experiments. The results of our study indicate that laryngeal exocrine secretion and blood flow are regulated by postganglionic autonomic parasympathetic fibers from intralaryngeal ganglia that contain VIP alone or VIP with ENK and/or SP, and that these ganglionic neurons may be innervated by CGRP-IR extrinsic nerve fibers.     

Distribution of neuropeptide-like immunoreactive nerve fibers in the canine larynx.

The distribution of neuropeptide immunoreactive nerve fibers in the canine larynx was examined. In the epithelium of supra- and subglottic regions, a dense distribution of substance P (SP)- and calcitonin gene-related polypeptide (CGRP)-immunoreactive (IR) nerve fibers was observed. Some vasoactive intestinal polypeptide (VIP)-IR intraepithelial nerve fibers were also seen in the subglottic region. In the laryngeal glands, a dense distribution of VIP-IR nerve fibers with a few SP- and enkephalin (ENK)-IR nerve fibers were found around the acini. In the walls of arteries in the lamina propria, many VIP-, SP-, and CGRP-IR nerve fibers were seen, whereas neuropeptide Y-, ENK-, and VIP-IR nerve fibers were predominantly distributed around the arteries in the vocal muscle. In the free edge of the vocal cord, few immunoreactive nerve fibers were detected within the epithelium and around the arteries in the lamina propria. These results suggest that there are regional differences in the occurrence of peptides in nerve fibers innervating the epithelium and the blood vessels in the larynx and that the perception mechanism of the epithelium and the regulatory system of local blood flow are varied according to their location in the larynx.     

Immunohistochemical and enzyme histochemical localization of peptidergic, aminergic and cholinergic nerve fibers in the rat seminal vesicle.

The distribution of the autonomic nervous system in the rat seminal vesicle was studied with immuno- and enzyme-histochemistry. In the smooth muscle layer, a large number of neuropeptide Y (NPY)- and tyrosine hydroxylase (TH)-immunoreactive nerve fibers were observed, while few fibers were distributed in the mucosal layer. A small number of substance P-immunoreactive fibers were present only in the smooth muscular layer, but vasoactive intestinal polypeptide (VIP)-immunoreactive fibers were abundantly distributed in both layers. In the mucosal layer, the VIP-fibers ran attached to blood vessels and encircled the glandular cavities. Acetylcholinesterase-positive fibers were also observed in the mucosal and muscular layers. Electron-microscopic studies revealed that NPY- and TH-containing nerve terminals were in apposition to smooth muscle cells, and VIP-fibers terminated at blood vessels. These results suggest different modes of action of NPY-, TH- and VIP-containing nerve fibers in the seminal vesicle.     

不同年龄段牙髓组织比例及牙髓干细胞分布的实验研究

目的通过观察不同年龄段前磨牙及第三磨牙牙髓占牙齿的比例,及干细胞分布情况,评估牙齿组织工程l临床适宜的选材标本。方法临床收集不同年龄段因正畸或其他原因拔除的无龋坏、无牙周疾病等的健康前磨牙及第三磨牙,分别称重牙齿及牙髓并计算比例。扫描电镜观察牙髓表面形态。脱钙后病理切片观察．牙髓组织内成牙本质细胞层完整与否,观察细胞与纤维成分比例;阿尔新蓝一希尔红染色,偏振光显微镜观察有无双折射现象：免疫组化染色观察．间充质干细胞特异性标记物STRO-1以及成牙本质相关蛋白牙本质涎蛋白DSP的表达情况．评估牙髓干细胞在牙髓组织中的分布情况。结果随着年龄的增长．前磨牙及第三磨牙牙髓占牙齿比例逐渐缩小．11～20岁年龄段与31～40岁、41～50岁,以及50岁以上年龄段比较．有统计学差异。扫描电镜下可见摘除的牙髓组织类似树桩．有些区域比较平滑,有些区域表面有不规则突起。牙髓组织内存在双折射。STRO-1阳性表达细胞在牙髓组织中散在分布．在血管周围较密集。DSP表达则主要集中在牙本质区域及成牙本质细胞内,牙髓内仅有少量表达。结论随着年龄的增长．牙髓逐渐萎缩,牙髓干细胞的分布集中于血管周围．在牙髓组织内散在分布．未分化的牙髓干细胞不表达成牙本质相关蛋白DSP。     

Distribution and property of nerve fibers in human long bone tissue

Objective： To observe the distribution of the nerve fibers in the bone tissue and the entry points of these fibers into the bone. Methods： The adult tibia was used for the ground sections which were afterwards made into the slice sections by decalcification in ethylenediamine tetraacetic acid （EDTA）. The ground sections were stained in silver and the slice sections were stained in silver and haematoxylin and eosin （HE） respectively. Then, the samples of the transmission electron microscope and the atomic force microscope were made and observed. Results ： In the human long bone tissue, many nerve fibers were distributed in the membrane, cortical bone, cancellous bone and marrow. The nerve fibers entered the bone from the nutrient foramen, and passed through the nutrient canal, Haversian＇ s canal and Volkmann＇ s canal, and finally into the bone marrow. In the nutrient canal, the nerve fibers, mainly the medullary nerve fibers, followed the blood vessel into the bone. In the cortical bone, the nerve fibers also followed the blood vessels and were mainly distributed along Haversian＇ s canal and Volkmann＇ s canal. In the bone trabecular and bone marrow, there were many nerve fiber endings arranged around the blood vessels, mainly around the tunica media of medium-size arteries in the marrow and around capillary blood vessels, and a few scattered in the bone marrow. There were sporadic nerve endings in epiphyseal plate and no nerve fibers permeated epiphysis to diaphysis. No distribution of nerve fibers could be found in cartilaginous part. Conclusions： There are many nerve fibers in bone and the nerve passageway is nutrient foramen, Volkman＇ s canal, Haversian＇ s canal and bone marrow.     

烧伤大鼠小肠肌间神经丛中一氧化氮合酶的组织化学研究

目的　探讨烧伤后大鼠小肠壁内一氧化氮合酶 (NOS)的活性及分布的变化规律。　方法　采用NADPH -黄递酶 (NDP)组织化学法和整装铺片技术对烧伤大鼠小肠壁内NOS的活性及分布进行定量和定位研究。　结果　NOS广泛分布于大鼠小肠壁内 ,主要定位于肌间神经丛。NOS阳性神经元多为圆形和卵圆形 ,神经纤维中多含有膨体 ,形成“串珠”样结构 ,且多与血管和肌纤维伴行。烧伤后肌间神经丛中NOS阳性神经元密度变化不显著 (P >0 .0 5 ) ,但神经元内NOS活性则显著下降(P <0 .0 5～ 0 .0 1) ,同时发现烧伤后NOS阳性神经元结构模糊 ,神经纤维断裂较多 ,未能形成完整的神经纤维网 ,神经纤维中NOS阳性膨体亦明显减少。结论　烧伤后大鼠小肠肌间神经丛中NOS阳性神经结构受损 ,神经元内NOS活性下降 ,同时一氧化氮 (NO)的释放途径亦存在障碍 ,这些变化可能与烧伤后肠道结构受损 ,功能障碍密切相关     

Alteration in gastric nerve fibers containing gastrin-releasing peptide in relation to the gastrin-producing cell population after truncal vagotomy in a rat model

The fine structural alteration in the gastric nerve fibers containing gastrin-releasing peptide (GRP) was studied in relation to the dynamics of gastrin-producing cells (G-cells) after truncal vagotomy in a rat model. The circulating gastrin levels were markedly elevated from the 1st day after vagotomy and the number of G-cells with positive immunoreaction for G17 and G34(1–15) was significantly increased in the vagotomized group. On the 3rd day after vagotomy, the G-cells showed conspicuous ultrastructural changes characterized by hypertrophy of the Golgi complexes and increased numbers of secretory granules. The GRP-positive nerve fibers formed a fine network in the gastric wall and were densely distributed in the oxyntic mucosa close to the blood vessels and showing varicosites composed of either small clear or GRP-positive large vesicles containing an electron-dense core. In the oxyntic mucosa of the vagotomized rats, axonal swelling of the nerves occurred on the 3rd day, and a depletion of GRP immunoreactivity was evidenced by a markedly decreased number of large-cored vesicles on the 7th day, when the serum GRP levels were also found to be markedly elevated. These findings indicate that the alteration in gastric nerve fibers containing GRP after truncal vagotomy may be related to hypergastrinemia and antral G-cell hyperplasia in the rat gastric mucosa.     

NK1受体在大鼠牙髓及牙周组织中的表达及意义

目的:观察NK1受体在大鼠牙髓及牙周组织中的表达情况。方法:采用冰冻切片和ABC免疫组化方法,检测NK1受体在大鼠牙髓及牙周组织中的表达。结果:NK1受体在大鼠牙周及牙髓组织内分布广泛,在牙周膜成纤维细胞、成牙骨质细胞、成牙本质细胞均呈强阳性表达;在牙髓成纤维细胞和牙槽骨细胞呈弱阳性表达;在牙齿根髓和冠髓内可发现染色阳性的血管样条索。结论:P物质可能直接作用于牙髓和牙周组织,细胞在牙髓及牙周炎症、修复反应中发挥作用。     

Neuroendocrine innervation of the hepatic vessels in the rat and in man

An extensive array of nerve fibers ramify around the afferent blood vessels of the liver and the extrahepatic and intrahepatic biliary pathways, and are thought to be involved in regulation of blood flow. Although the role of sympathetic innervation is established, little is known about the location or role of regulatory peptidergic innervation in the liver. We examined the anatomic distribution of a wide variety of regulatory peptides and several neural antigens by in situ immunohistochemistry in the rat and in man. A rich peptidergic plexus of nerve fibers and ganglion cells was observed around the arterial vessels in both species, with intense immunoreactivity for neuron-specific enolase, neurofilaments, neuropeptide Y, substance P, and vasoactive intestinal polypeptide. S-100 protein immunoreactivity was seen principally in large nerve bundles, suggesting that the majority of nerves in this area were unmyelinated. In contrast, the portal vessels revealed very little peptidergic innervation. No staining was observed with antibodies directed against insulin, glucagon, gastrin, serotonin, met-enkephalin-Arg-Gly-Leu, cholecystokinin, or growth hormone. These findings indicate the presence of a rich, although selective, peptidergic plexus surrounding afferent hepatic blood vessels. This plexus may play an important role in regulation of hepatic blood flow.