先天性巨结肠肌间神经丛neurexin和neuroligin蛋白的表达及意义

目的：观察先天性巨结肠肌间神经丛neurexin、neuroligin蛋白在狭窄段、移行段及扩张段的表达及意义。方法：将20例先天性巨结肠狭窄段、移行段和扩张段制备成肌间神经丛铺片标本,采用免疫组织化学技术,对neurexin、neuroligin蛋白进行检测。结果：neurexin、neuroligin蛋白在扩张段表达最多,移行段明显减少,狭窄段无表达（P〈0.05）;同一组织neurexin蛋白和neuroligin蛋白的表达差异无统计学意义（P〉0.05）。结论：neurexin、neuroligin蛋白在人体肠神经系统肌间神经丛中表达的减少及缺失是导致先天性巨结肠肠蠕动功能丧失的重要原因。     

成人型巨结肠与慢性便秘

成人型巨结肠分为先天性和后天获得性两大类，前者是由于远端结肠、直肠黏膜下神经丛（Meissner神经丛）和肌间神经丛（Auerbach神经丛）中缺乏神经节细胞所致，以新生儿和婴幼儿多见，约10％病人出生后症状轻微，直到成人期症状明显时才得以诊断；而后者则可由精神性、神经性、药物性、     

升结肠翻转术治疗先天性巨结肠症(附28例报告)

<正> 先天性巨结肠是一种常见的消化道畸形,其结肠肌间神经节细胞先天性缺如,病变肠段失去正常蠕动及排便反射的功能,其中长段型约占10％～40％,无神经节细胞段可达到降结肠及横结肠。而部分普通型患儿因就诊时间晚,近端结肠代偿性扩张肥厚严重,且肌间神经节细胞也继发变性。巨结肠根     

NSE和S-100抗体免疫标记在先天性巨结肠病理诊断中的意义

先天性巨结肠(megacolon,又名Hirschsprung病)是常见消化道发育畸形,病理学改变为神经节细胞缺乏症,系直肠和下段结肠平滑肌神经丛缺乏神经节细胞,导致肠管痉挛正常蠕动消失形成功能性肠梗阻。先天性巨结肠的病理学诊断依靠全层肠标本活检,但常规HE切片光镜下难以识别神经节细胞,尤其是在新生儿;乙酰胆碱酯酶特殊染色可显示固有膜和黏膜肌层乙酰胆碱酯酶阳性纤维明显增加,但它作为辅助诊断应用存在争议,假阳性和假阴性反应曾有报告[1]。VinoresSA,MayE认为免疫组化标记神经元特异性烯醇化酶(NSE)非常有利于证实神经节细胞的存在[2]。本…     

慢传输型便秘大鼠结肠肌间神经丛内VIP能神经、SP能神经的免疫组化研究

目的　探讨结肠肌间神经丛血管活性肠肽 (VIP)能神经及P物质 (SP)能神经在慢传输型便秘发病中的作用。方法　建立大鼠慢传输型便秘模型 ,利用铺片技术制作结肠肌间神经丛标本 ,采用免疫组织化学法在铺片上显示便秘大鼠结肠肌间神经丛内VIP能神经及SP能神经的分布及形态学改变。结果　与正常大鼠比较 ,便秘大鼠结肠肌间神经丛VIP能神经节及节间束细小 ,节内神经元数量明显减少 ,节间束内神经纤维分布稀疏 (平均面密度值 2 3.0 8± 1.82vs 19.35± 1 13,P <0 0 1) ;便秘大鼠结肠肌间神经丛SP能神经节明显增大 ,神经束增粗 ,神经元和神经纤维明显增多 (平均面密度值 15.4 5± 1 0 5vs18.14±1 18,P <0 .0 1)。结论　慢传输型便秘大鼠结肠壁内存在明显的神经病理学改变 ,结肠传输功能障碍可能与结肠肌间神经丛VIP和SP能神经病理改变和 或功能障碍有关。     

结肠冗长的特发性便秘的结肠形态病理学分析

目的 研究表现为结肠冗长的特发性便秘的结肠形态病理学改变。方法 选取14例原发性结肠冗长病例与10例不伴便秘与梗阻的结肠癌病例作为对照,通过肉眼及HE染色后显微镜观察和图像分析,比较两组的异同点。并通过定量测量和统计学分析得出显著性结果。结果 原发性结肠冗长病例外观正常,显微镜下突出的改变为5例肌间神经丛中可见巨大的神经节细胞。图像分析结果显示其粘膜下血管面积、神经丛面积和神经节细胞数,肌间神经丛面积和神经节细胞数均显著大于对照组,肌层厚度则无显著性差异。结论 原发性结肠冗长有其特征性的神经病理学改变,进一步支持了特发性便秘是一种神经系统异常性疾病的论断。     

先天性巨结肠的外科治疗

先天性巨结肠（Hirschsprungdisease，HD）是以便秘为主要临床表现的一种常见的消化道畸形．病因为外胚层神经嵴细胞迁移发育过程停顿．病变肠壁肌间神经丛和黏膜下神经丛的神经节细胞缺如。使肠段失去正常蠕动（即间歇性收缩和松弛的推进式运动）而处于经常的痉挛状态，造成粪便排出障碍．近端肠管则被动继发扩张，异常扩大、肥厚，形成所谓的巨结肠。     

先天性巨结肠不同节段肠壁神经、平滑肌的分布及临床意义

目的 观察先天性巨结肠(HD)不同节段肠壁神经和平滑肌的病变范围,探讨先天性巨结肠根治术后肠动力功能紊乱原因及手术切除结肠范围.方法 用免疫组织化学和苏木素-伊红(HE)染色法,检测20例先天性巨结肠肠壁神经节细胞、神经纤维和平滑肌细胞病理组织学改变及分布范围.结果 巨结肠不同节段肠壁神经节细胞、神经纤维数量及突触素(Syn)、神经节细胞黏附分子(NCAM)的阳性表达,在距扩张远端8 cm虽未达到正常,但与对照组差异减小(P＞0.05).环肌层和纵肌层出现不同程度增厚,在距扩张远端8 cm仍未正常(P＜0.01).肌层出现空泡样变,与对照组比较差异无统计学意义(P＞0.05).结论 先天性巨结肠切除段肠壁神经、平滑肌层均存在病变,在距扩张段的远端8 cm处,两者病变总体缓解.在允许情况下,手术切除结肠的范围应达到或超过此范围.     

神经元特异性烯醇化酶和银染色在先天性巨结肠类缘病诊断中的应用

为探讨先天性巨结肠类缘病的病理诊断,分析了20例类缘病NSE及银染色。结果表明,NSE结合银染色对肌间和粘膜下神经丛中神经节细胞、神经纤维有特征性表达。认为NSE结合银染色是一种诊断先天性巨结肠类缘病有价值的诊断方法。     

神经生长相关蛋白-43基因及其蛋白在先天性巨结肠肠组织中的表达

目的探索神经生长相关蛋白-43(GAP-43)在先天性巨结肠痉挛段和扩张段肠组织中的表达情况,进一步探索先天性巨结肠的发病机理。方法收集2012年1月至2013年6月期间因先天性巨结肠于深圳市儿童医院行巨结肠根治术的30例患儿的活体肠组织标本,行免疫组化染色及荧光定量聚合酶链反应(RT-PCR)以检测扩张段和狭窄段肠组织中GAP-43 m RNA及其蛋白的表达,并比较扩张段和狭窄段肠组织中两者表达的差异。结果RT-PCR结果显示,痉挛组肠组织中GAP-43 m RNA的表达水平的中位数为0.052 8,低于扩张组的0.119 0(P<0.05)。免疫组化染色结果显示:30例患儿的狭窄段肠组织和扩张段肠组织的肌间神经丛及黏膜下神经丛中GAP-43蛋白均呈阳性表达,但痉挛段黏膜下神经丛和肌间神经丛的染色均较扩张段相应部位浅;与扩张组的黏膜下神经丛和肌间神经丛比较,痉挛组相应部位的GAP-43蛋白的平均光密度值均较低(P<0.05)。结论先天性巨结肠狭窄段肠组织中GAP-43蛋白的表达较扩张段肠组织下调,提示GAP-43蛋白可能是先天性巨结肠发病的危险因素之一。     

Reduced Neuronal Innervation in the Distal End of the Proximal Esophageal Atretic Segment in Cases of Esophageal Atresia with Distal Tracheoesophageal Fistula

Background Esophageal dysmotility is a common occurence after surgical repair of proximal esophageal atresia (EA) and distal tracheoesophageal fistula (TEF). The etiology of this motility disorder, however, remains controversial. Esophageal dysmotility also is present in isolated TEF or EA before surgery, suggesting a congenital cause. However, there is no information available in the literature with regard to the intramural nervous system of the human esophagus in EA-TEF. Patients and Methods We examined the distal end of proximal esophageal atretic segment of neonates undergoing EA-TEF repair for intrinsic neuronal innervation. Using specific antibodies, we studied neuronal markers of specimens from nine cases of EA-TEF and 9 cases of normal esophagus by immunohistochemistry using neurofilament (NF), synaptophysin (SY), S100, and glial cell line-derived neurotrophic factor (GDNF). Results In the atretic segment, specimens staining with hematoxylin and eosin showed that there were marked hypoganglionosis and immature ganglion cells in the myenteric plexus. GDNF immunoreactivity in the atretic esophagus were markedly reduced in both the muscular layer and myenteric plexus. SY and NF-immunorective nerve fibers were distributed throughout the myenteric plexus of the normal esophagus, but the scarcity of these immunoreactive nerve fibers in the atretic esophagus was apparent. In contrast, the density of immunorective nerve fibers for S100 in the myenteric plexus and muscular layer was increased in the distal end of the atretic esophagus. Conclusion We concluded that the distribution of ganglion cells and some nerve fibers in the distal end of the atretic esophageal segment is deficient. Inadequate and abnormal neuronal innervation of the esophagus could be related to the esophageal dysmotility seen in EA. Because GDNF is a survival factor for central and peripheral neurons, defective expression of GDNF could have an important role in the defective and/or abnormal neuronal innervation of atretic esophageal segment.     

Three-dimensional morphology of gut innervation in total intestinal aganglionosis using whole-mount preparation

BACKGROUND: Total intestinal aganglionosis (TIA) is a rare form of Hirschsprung's disease (HD). The aim of this study was to examine the 3-dimensional morphology of the myentric plexus of the entire gastrointestinal tract in a newborn with total intestinal aganglionosis. METHODS: Whole-mount preparations were made of the entire gastrointestinal tract using NADPH-diaphorase histochemistry and c-kit (a marker of interstitial cells of Cajal) immunohistochemistry. RESULTS: Whole-mount preparations of the esophagus, stomach, and duodenum showed 3-dimensional morphology of the myenteric plexus forming a meshlike network of nerve fibers, connected to each other and to ganglia. There were large numbers of NADPH-diaphrase-positive nerve fibers between the muscle fibers in the circular muscle layer. In esophagus, stomach, and duodenum, c-kit-positive interstitial cells of Cajal (ICC) formed a 3-dimensional network between the two muscle layers and also were abundant within the circular muscle layer. In the jejunum, ileum, and colon, the myenteric plexus was absent and was replaced by hypertrophic nerve bundles that stained weakly with NADPH-diaphrase. Circular muscle layer completely lacked NADPH-diaphrase-positive nerve fibers. The c-kit-positive ICCs in the jejunum, ileum, and colon were sparse and localized mainly around the nerve trunks between the circular and longitudinal muscle layers. CONCLUSIONS: Whole-mount preparation is an elegant 3-dimensional technique in which the relationship of branching and interconnecting nerve fibers to each other and to muscle can be seen clearly. Absence of myenteric plexus, lack of nitrergic innervation, and depletion of interstitial cells of Cajal in the bowel wall throughout the small and large bowel contribute to the inability of the smooth muscle to relax, thereby causing lack of peristalsis in TIA.     

Deficient neurogenic innervation of the myenteric plexus with normal submucous plexus involving the entire small and large bowel

A newborn presented with a picture of intestinal obstruction. Multiple biopsies of the small and large bowel showed an unusual neurogenic innervation. The myenteric plexus of Auerbach was severely depleted of ganglion cells and nerve fibers, while the submucous plexus of Meissner was normally innervated. An ileostomy failed to function and extended trial with experimental smooth muscle stimulant (Cisapride) was equally ineffective. The patient was finally treated by a myectomy from the duodenum to the descending colon with a sigmoid colostomy. This procedure, coupled with a Nissen fundoplication, stopped the vomiting and allowed normal defecation through the colostomy. The patient is presently taking increasing increments of oral fluids with a concomitant decrease in the volume of parenteral nutrition. The myectomy initiated marked hypertrophy of the muscularis mucosa. Could this muscular hypertrophy account for the improvement in bowel function? Possible etiology will be discussed. We caution that rectal submucosal suction biopsy alone may be misleading if normal ganglion cells and nerve fibers are found, yet the patient's clinical symptoms fail to improve. A full thickness bowel wall biopsy is then recommended.     

Immunohistochemical characterization of abnormal innervation of colon in Hirschsprung's disease using D7 monoclonal antibody

Innervation patterns in normal and aganglionic colon were studied using a panel of antineuronal cell antibodies. One antibody, D7, which recognizes a subset of neuronal cells of the peripheral and central nervous system reacted strongly with nerve fibers in the circular muscle of the normal colon. Immunohistochemical scanning of the entire resected specimen of colon from three children with Hirschsprung's disease demonstrated large numbers of D7 immunoreactive nerve fibers in the circular muscle of the ganglionic colon, few fibers in the transitional zone, and no immunoreactive fibers in the aganglionic segment of bowel. While the absence of D7 immunoreactive fibers paralleled the absence of myenteric ganglion cells in the aganglionic segment, a critical region of colon was identified wherein D7 reactive fibers were evident ahead of the appearance of ganglion cells. These findings indicate that the fundamental pathology in Hirschsprung's disease is not only the absence of ganglion cells of the myenteric and submucuous plexuses but also the absence of D7 immunoreactive fibers in the circular muscle of the colon.     

Segmental dilatation of sigmoid colon in a neonate: atypical presentation and histology

Segmental dilatation of the colon is a rare disorder of colonic motility in children, often presenting with severe constipation in older infants, children, and occasionally adults. It may mimic the commoner Hirschsprung disease clinicoradiologically but differs in that the ganglion cell morphology and distribution are typically normal in the colon. We report a neonate with segmental dilatation of the sigmoid colon who had an atypical clinical presentation and describe certain abnormalities in bowel histology (hypertrophied muscularis propria, nerve plexus, and ganglion cells located within the circular layer rather than the normal myenteric location), for the first time in the English literature.     

Hypoganglionosis of the myenteric plexus with normal Meissner's plexus: a new variant of colonic ganglion cell disorders

A 16-year-old girl with a history of achalasia of the lower esophagus was investigated for progressive constipation since the age of five years. Normal ganglion cells of the Meissner's plexus were found in a mucosal rectal biopsy. Manometric studies showed absence of the normal rectosphincteric relaxation reflex. A long posterior rectal myectomy was performed in which no ganglion cells were found between the muscular layers. The patient underwent a rectosigmoid resection with a low anastomosis according to Duhamel. In the resected bowel, a normal Meissner's plexus was again found while marked hypoganglionosis of the myenteric plexus was evident. Intranuclear inclusion bodies in Schwann cells and scattered inflammatory cells were noteworthy. This case may be an example of an acquired hypoganglionosis localized to the myenteric plexus.     

Morphological alterations in small intestine of rats with myenteric plexus denervation

We aimed to investigate the effect of myenteric denervation by benzalkonium chloride (BAC) on small intestine morphology in the rat, and whether segmental myenteric denervation alters morphology elsewhere in the small intestine. Forty male Sprague-Dawley rats were equally divided into 4 groups: control (0.9% NaCl); denervation (0.062% BAC); chemical inflammation (5% acetic acid), and intraluminal stasis produced by partial obstruction. 28 days after operation tissue samples were taken from the treated segment, 10 cm distal to the treated segment, and 20 cm proximal to the treated segment. Morphological changes and the number of ganglion cells were examined under the light microscope. BAC application reduced the number of myenteric neurons by 85% in the treated segment. Denervation increased villus height and crypt depth in the treated and proximal segments. But changes in muscle thickness were seen throughout the intestine. As a result, although myenteric plexus denervation caused mucosa morphology in the treated and proximal segments, it caused smooth muscle changes throughout the small intestine.     

Colonic nerve network demonstrated by quinacrine

Quinacrine can be used for fluorescence microscopy to visualize certain cell bodies and fibres in the myenteric plexus of the intestine, forming a network different from the cholinergic and adrenergic systems. In the Piebald mouse, animal model of congenital megacolon, the quinacrine positive plexus is thin in the distal narrowed segment and well developed in the large proximal zone. In Hirschsprung's disease, the quniacrine positive network has the same pathologic features but, contrary to the animal model, there is an increased number and size of extrinsic cholinergic and adrenergic fibres. So this mouse is an animal model of a pseudo-Hirschsprung's disease, which has not yet been reported in human pathology.     

The pathogenesis of pseudoachalasia: a clinicopathologic study of 13 cases of a rare entity

Pseudoachalasia is an esophageal motor disorder usually associated with malignancy that has clinical, radiographic, and manometric findings that are often indistinguishable from primary achalasia. There are few reports examining the histologic features of the associated neoplasms and their relationship with the esophageal myenteric plexus. We studied the clinical and pathologic features of 13 cases of pseudoachalasia seen at our institution between 1979 and 1999. Detailed clinical and radiographic data were obtained from medical records. In all cases available histologic material was reviewed to confirm the presence and type of associated neoplasm. When possible, the relationship of the neoplasm to the esophageal myenteric plexus was examined. In selected cases immunohistochemical stains were performed to further evaluate this relationship. All patients had clinical, radiographic, and manometric features similar to primary achalasia. The cohort included seven men and six women, age range 24-79 years (median 61 years). Associated neoplasms included esophageal adenocarcinoma arising in Barrett's esophagus (n = 1), adenocarcinoma of the esophagogastric junction (n = 7), metastatic renal cell carcinoma to the esophagogastric junction (n = 1), breast adenocarcinoma (n = 1), pulmonary small cell carcinoma (n = 1), pleural malignant mesothelioma (n = 1), and mediastinal fibrosis (n = 1). The mechanism of pseudoachalasia was consistent with neoplastic infiltration of the esophageal myenteric plexus in 11 cases. Neoplastic cells surrounded myenteric ganglion cells, which appeared normal in number and morphology. In the patient with pulmonary small cell carcinoma, there was no evidence of neoplastic infiltration of the esophagogastric junction, and anti-ANNA-1 antibody was detected, suggesting a paraneoplastic syndrome. Tissue obtained at the time of esophagomyotomy revealed lymphocytic myenteric inflammation and marked depletion of ganglion cells identical to that seen in primary achalasia. The mechanism pseudoachalasia in the patient with breast adenocarcinoma is uncertain, as there was no evidence of direct involvement of the esophagogastric junction. In summary, we describe 13 cases of pseudoachalasia resulting in a clinical syndrome indistinguishable from primary achalasia. The most common mechanism is direct involvement of the esophageal myenteric plexus by neoplastic cells. Rarely, a distant neoplasm may cause this syndrome as a paraneoplastic process.     

Diabetic autonomic neuropathy in BB rat. Ultrastructural and morphometric changes in parasympathetic nerves

Longitudinal electron-microscopic and morphometric studies of autonomic nerves containing predominantly parasympathetic fibers were undertaken in the spontaneously diabetic BB rat. Unmyelinated fibers of the diabetic vagus nerve and myelinated fibers of the penile nerve showed increased numbers of axonal glycogenosomes and axonal sequestration. Morphometric examination of myelinated and unmyelinated fibers of the vagus nerve revealed diminished fiber size compared with age-matched control animals. The distal myenteric nerve showed marked degenerative changes, whereas no structural changes could be demonstrated in intra-myenteric ganglion cells. These changes are similar to those described previously in somatic nerves of this model but different from those seen in sympathetic nerves of the diabetic BB rat.