组织蛋白酶D在先天性巨结肠症中的表达

先天性巨结肠（HD）是常见的小儿胃肠道先天畸形,其病因目前仍无定论,其基本病变是狭窄段肠壁黏膜下和肌间神经丛内缺乏神经节细胞.识别黏膜下神经丛内未成熟神经节细胞,有可能成为诊断本病的理论依据.本文通过观察组织蛋白酶D（CD）在HD无神经节细胞肠段的分布情况,试图寻找神经节细胞的标志物,并将此技术用于HD诊断.     

先天性巨结肠症C-fos原癌基因表达的研究

目的探讨C-fos原癌基因与先天性巨结肠症发病机制间的关系。方法采用RT-PCR技术及免疫组织化学SP方法对巨结肠无神经节细胞的巨结肠痉挛段以及有神经节细胞的扩张段、正常肠组织各12例标本检测C-fos原癌基因mRNA的表达和观察其蛋白免疫组化状况。结果无神经节细胞的巨结肠痉挛段C-fosmRNA表达水平明显高于有神经节细胞扩张段近端(痉挛段相对含量2．11±0．09,扩张段相对含量0．55±0．06,单位mass),巨结肠扩张段近端与正常肠组织相比无明显差异;C-fos蛋白免疫组化巨结肠无神经节细胞的痉挛段粘膜固有层和粘膜肌层中出现大量的棕褐色C-fos蛋白表达产物,而巨结肠近段的扩张段未见C-fos蛋白表达阳性产物。结论C-fos基因可能参与了神经细胞的损害。C-fosmRNA表达及C-fos蛋白表达异常可能参与先天性巨结肠的发病机制。     

肠神经节细胞瘤病——神经生长因子产生过度的表现?

多发性内分泌瘤(MEN)Ⅱb综合征的完整临床表现包括嗜铬细胞瘤、甲状腺髓癌、粘膜神经瘤和顽固性便秘,类似巨结肠(Hirshsprung病)的临床和X线表现并不少见。但与该病的无神经节不同,肠肌神经丛大片增生,故被称为神经节细胞瘤病,发生机理还不清楚。本文报告的病变超微结构有不典型“侵入”型神经增殖,同时有相对多的充满小颗粒的神经末梢,表明有一个神     

慢传输型便秘肠神经系统一氧化氮合酶和P物质的分布意义

目的研究结肠一氧化氮（ＮＯ）和Ｐ物质（ＳＰ）在慢传输型便秘（ＳＴＣ）发病中的作用．方法应用一氧化氮合酶（ＮＯＳ）和ＳＰ的兔多克隆抗体，对手术切除的１５例ＳＴＣ患者（男２例，女１３例，年龄２６岁～６８岁）和１１例对照组患者乙状结肠标本进行免疫组化染色和半定量分析．结果ＳＴＣ患者乙状结肠肌间神经丛ＮＯＳ免疫反应性明显升高（χ２＝１５６３，Ｐ＜００１），而ＳＰ免疫反应性明显降低（χ２＝１３４４，Ｐ＜００１）；粘膜下神经丛ＮＯＳ及ＳＰ免疫反应性与对照组相差不显著．结论ＳＴＣ结肠神经系统ＮＯＳ和ＳＰ免疫反应性的异常变化，可能是其结肠传输减慢的神经病理基础     

先天性巨结肠相关基因的研究进展

先天性巨结肠（HD）又称肠无神经节细胞症，是一种神经嵴细胞源性疾病和多基因遗传病，病因和发病机制目前尚不清楚。近年来新发现了数种与HD发病相关的基因，现综述如下。     

先天性巨结肠组织化学诊断的临床评价

先天性巨结肠(HD)的诊断一般是根据X线、肛门直肠测压和直肠活检.但X线表现在新生儿期不尽可靠;肛门直肠测压可显示内括约肌松弛反射缺如,对诊断有裨益,但有时难以取得患儿合作;Swenson式直肠全层活检需要用全麻,技术上有一定危险.近年发展了直肠吸吮活检组织化学染色法,是对诊断的一大进步,尤其对新生儿病例,且不需麻醉,罕有并发症.直肠活检染色检查乙酰胆碱酯酶(AChE)活力,HD时可在固有膜和粘膜肌层观察到显著的神经纤维,同时缺乏节     

复发型结肠黏膜-黏膜下拉长型息肉一例

1例腹痛4年后复发患者，结肠检查见多发息肉，病理提示腺体结构和腺上皮均无异型，黏膜肌层完整，黏膜下层见大量增生的厚壁血管及淋巴管沿息肉长轴方向走行，局部血管充血，免疫组化染色结果示增生的血管内皮细胞，CD31（+），最终诊断为结肠黏膜-黏膜下拉长型息肉。     

先天性巨结肠X线诊断和分析

目的：探讨先天性巨结肠（HD）的临床表现、X线表现及其分型，重点探讨X线钡灌肠的方法和漏诊原因。方法：对198例已经手术和粘膜活检证实的HD进行回顾性分析。结果：（1）HD的主要临床表现为出生后少或不排胎粪、呕吐及进行性腹胀，X线检查是诊断本病的主要检查方法。(2)根据病变痉挛段范围不同，可将本组病例分为五型。结论：钡灌肠检查是诊断HD最可靠、最简便的方法。     

HD的病因,胚胎及分子生物学

ＨＤ的病因、胚胎及分子生物学滨州医学院附属医院（２５６６０３）滨州医学院免疫学教研室傅廷亮王运平先天性巨结肠（ＨＤ）的特征是结肠远段神经丛中神经节细胞先天性缺如。尽管ＨＤ的病因尚未完全阐明，但随着免疫组化和重组ＤＮＡ技术的应用，近年来有关ＨＤ病因的研...     

先天性巨结肠围手术期护理体会

先天性巨结肠（HD）是新生儿消化道梗阻的常见原因之一,其发病机制为结肠远端及直肠缺乏神经节细胞而造成消化道梗阻,70％-90％的患儿可在新生儿时期明确诊断。手术切除可以取得良好效果,正确的围手术期护理也是手术成功的重要环节。我们对42例HD患儿围手术期采取多种护理措施,取得了良好的效果,现将护理体会报告如下。     

Intestinal neuronal dysplasia

Intestinal neuronal dysplasia type B (IND B) is currently defined as a disease of the submucous plexus of the intestine. The aetiology of IND B remains largely obscure. The congenital origin of IND B is supposed; nevertheless, the findings of IND B associated with chronic intestinal obstruction support the notion that this disease could be caused by a reaction of the enteral nervous system to intestinal obstruction or inflammatory disease either in the fetal or the postnatal period. The treatment of IND type B has no unified concept of treatment. The ultimate clinical diagnosis of IND B should be based on a definitive histological diagnosis relating to clinical symptoms, the course of treatment and long-term follow-up of patients with this dysfunction of intestinal motility, despite the fact that no correlations of the clinical picture, radiological investigation and anorectal manometric studies with IND B have been found so far.     

Morphometric quantification of normal submucous plexus in the distal rectum of adult healthy volunteers

OBJECTIVE: Inadequate morphometric characterization of the normal adult submucous plexus has precluded the diagnosis of colonic dysganglionoses associated with constipation, such as intestinal neuronal dysplasia type B (IND B). The internal submucous plexus (Meissner plexus) was morphometrically quantified in adult healthy volunteers. DESIGN: Open, prospective morphometric study in balanced groups of female and male volunteers. PARTICIPANTS: Thirty-seven adult healthy male and female volunteers with normal bowel function and no history of gastrointestinal disease. METHODS: Four jumbo rectal biopsies (3-5 mm3) were taken 5 and 10 cm above the pectinate line. Two expert gastrointestinal pathologists assessed biopsy sections after specific nerve cell staining for lactic dehydrogenase, nitric oxide synthase and acetylcholinesterase, mainly for characteristics of ganglia and nerve cells in the submucous plexus. RESULTS: No healthy individual demonstrated over 20% of submucosal ganglia as giant ganglia or more than four giant ganglia per 30 sections (the morphometric criteria for IND B). Single submucosal nerve cells and ganglion numbers halved between 10 and 5 cm above the pectinate line, but there were no age or gender differences. The biological variability of nerve cell and ganglion density in the submucous plexus was large. CONCLUSIONS: Healthy adults show less than 20% of submucosal ganglia as giant ganglia and no more than four giant ganglia per 30 rectal biopsy sections. There is therefore no overlap with the histomorphological criteria of IND B. These data therefore support the specificity of the previously defined criteria for IND B in adults.     

Analysis of the RET, GDNF, EDN3, and EDNRB genes in patients with intestinal neuronal dysplasia and Hirschsprung disease

BACKGROUND: Hirschsprung disease (HSCR) is a frequent congenital disorder with an incidence of 1 in 5000 live births, characterised by the absence of parasympathetic intramural ganglion cells in the hindgut resulting in intestinal obstruction in neonates and severe constipation in infants and adults. Intestinal neuronal dysplasia (IND) shares clinical features with HSCR but the submucosal parasympathetic plexus is affected. IND has been proposed as one of the most frequent causes of chronic constipation and is often associated with HSCR. METHODS: We examined 29 patients diagnosed with sporadic HSCR, 20 patients with IND, and 12 patients with mixed HSCR/IND for mutations in the coding regions of the RET, GDNF, EDNRB, and EDN3 genes. The entire coding regions were analysed by single strand conformational polymorphism and DNA sequencing. RESULTS: Only three RET mutations were detected in patients with HSCR. In patients with IND or a mixed HSCR/IND phenotype, no mutations in these genes were observed. While HSCR and HSCR/IND showed over representation of a specific RET polymorphism in exon 2, IND exhibited a significantly lower frequency comparable with that of controls. CONCLUSIONS: The mutation frequency found in our sporadic HSCR patients (10%) and the allelic distribution of RET polymorphisms are comparable with earlier published data. A significantly different allelic distribution in an established HSCR associated polymorphism argues against common genetic pathways for HSCR and IND.     

Histological studies on Hirschsprung's disease and its allied disorders in childhood

BACKGROUND/AIMS: To obtain accurate diagnosis for Hirschsprung's disease (HD) and its allied disorders such as hypoganglionosis (Hypo) and intestinal neuronal dysplasia (IND) in childhood patients with chronic constipation, we studied the histology of childhood patients with refractory constipation accompanied by abdominal distension and pain. METHODOLOGY: Based on clinical signs and symptoms noted on admission, all of 109 patients (60 males and 49 females, aged 2-15 years with a mean age of 9.8 years) were suspected to have chronic refractory constipation. To obtain accurate histological diagnosis in childhood patients with chronic refractory constipation, we performed rectal biopsies on these patients. Tissue samples were frozen and 12-microm sections were stained with acetylcholinesterase (AChE) by the method of Karnovsky and Roots, and with nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase by the modified Scherer-Singler's method. RESULTS: On the basis of histological studies using rectal biopsies, 20 cases were diagnosed with Hypo, 5 with HD, 2 with intestinal neuronal dysplasia (IND), and 82 with normal findings. The incidence of normal cases was significantly greater than that of allied disorders of HD including both Hypo and IND (P<0.0001). The incidence of Hypo was also significantly greater than that of Hypo and IND (P<0.01, P<0.0001, respectively). Both HD and IND could be diagnosed by rectal mucosal biopsies with AChE staining. However, accurate diagnosis of Hypo could be made only through examination of the submucosal and myenteric plexuses by NADPH-diaphorase staining in full-thickness rectal specimens. CONCLUSIONS: We were able to obtain accurate diagnosis of childhood patients with HD and IND by rectal mucosal biopsy with AChE staining. On the other hand, accurate histological diagnosis in patients with Hypo could also be obtained by NADPH-diaphorase staining in full-thickness rectal specimens. That is to say, it is easier for the investigator to detect the cholinergic fiber and ganglion cell in the gut wall using NADPH-diaphorase staining than by using AChE staining.     

Rectal biopsy for diagnosis of intestinal neuronal dysplasia in children: a prospective multicentre study on interobserver variation and clinical outcome

BACKGROUND: Intestinal neuronal dysplasia (IND) of the colonic submucous plexus is considered to be a congenital malformation of the enteric nervous system causing symptoms resembling those of Hirschsprung's disease. In contrast with the established diagnosis of aganglionosis using enzyme histochemistry, controversy exists over the diagnostic criteria of IND on rectal biopsies previously defined by a consensus report and the causal relation between morphological findings and clinical symptoms. AIMS: The interobserver variability was prospectively investigated with respect to final diagnoses and several histological features in rectal biopsy specimens from children suspected of having colonic motility disturbances. METHODS: 377 biopsy specimens from 108 children aged 4 days to 15 years were independently coded without knowledge of clinical symptoms by three experienced pathologists for 20 histological features, and a final diagnosis was given for every case. Interobserver variation for the different items and the final diagnosis were analysed using Cohen's kappa statistic. Clinical data at biopsy and outcome after 12 months were related to morphological findings. RESULTS: The three pathologists agreed completely with respect to the diagnosis Hirschsprung's disease (kappa = 1), but in only 14% of the children without aganglionosis. In 15 (17%) of the 87 children without aganglionosis, at least one pathologist judged the case as normal, while another diagnosed IND. kappa values were close to the zero value expected by chance for the diagnoses normal and IND. Young age was related to the presence of several morphological features-for example, acetylcholine esterase staining and presence of giant ganglia. Children with chronic constipation diagnosed as having IND, given no other specific diagnosis by any of the pathologists, were significantly younger (median 8.8 months) and had a higher cure rate after one year (60%) than constipated patients considered by all observers to have no histological abnormalities (median 6.1 years, cure rate 23%). CONCLUSIONS: In contrast with Hirschsprung's disease, there is a high interobserver variation with regard to the different morphological features and final diagnosis of IND, based on the criteria and conditions of the previous consensus report. The high frequency of histological "abnormalities" in young infants suggests that some of the features may represent a normal variant of postnatal development rather than a pathological process. Investigations using more refined and morphometric methods in rectal specimens from infants and children without bowel disease are needed to define the normal range of morphological appearance at different ages. These preliminary data indicate that, with current knowledge, rectal biopsy for diagnostic purposes should only be performed in constipated children for diagnosis of Hirschsprung's disease.     

肺癌的特异性神经元抗核抗体-Hu检测

目的：了解Hu抗体对肺癌的诊断意义。方法：用亲和免疫组织化学方法ABC法及免疫印迹技术测定24例患血清中的Hu抗体。结果：7例患血清中检出Hu抗体，小细胞肺癌6例，阳性率达66．7％。结论：Hu抗体检测对肺癌的早期诊断有较高应用价值。     

Abnormalities of the enteric nervous system in heterozygous endothelin B receptor deficient (spotting lethal) rats resembling intestinal neuronal dysplasia

BACKGROUND: A homozygous mutation of the endothelin B receptor (EDNRB) gene in spotting lethal (sl/sl) rats leads to Hirschsprung's disease (HSCR) with long segmented aganglionosis. However, the effects on the development of the enteric nervous system (ENS) promoted by a heterozygous mutation of the EDNRB gene are not known. The present study aimed to describe and morphometrically assess the phenotypic abnormalities of the ENS in heterozygous (+/sl) EDNRB deficient rats in comparison with homozygous (sl/sl) EDNRB deficient and wild-type (+/+) rats. METHODS: The distal small intestine, caecum, and colon were obtained from sl/sl, +/sl, and +/+ rats. To demonstrate the three dimensional organisation of the ENS, the intestinal wall was microdissected into wholemounts and incubated against the pan-neuronal marker protein gene product 9.5. Assessment of the ENS included morphometric quantification of ganglionic size and density, the number of nerve cells per ganglia, and the diameter of nerve fibre strands within both the myenteric and submucous plexus. RESULTS: Sl/sl rats were characterised by complete aganglionosis resembling the same histopathological features observed in patients with HSCR. +/sl rats revealed more subtle abnormalities of the ENS: the submucous plexus was characterised by a significantly increased ganglionic size and density, and the presence of hypertrophied nerve fibre strands. Morphometric evaluation of the myenteric plexus did not show statistically significant differences between +/sl and +/+ rats. CONCLUSIONS: In contrast with sl/sl rats, +/sl rats display non-aganglionated malformations of the ENS. Interestingly, these innervational abnormalities resemble the histopathological criteria for intestinal neuronal dysplasia (IND). Although IND has been described in several intestinal motility disorders, the concept of a clearly defined clinical-histopathological entity is still controversially discussed. The present findings support the concept of IND based on clearly defined morphological criteria suggesting a genetic link, and thus may provide a model for human IND. Furthermore, the data underline the critical role of the "gene dose" for the phenotypic effects promoted by the EDNRB/EDN3 system and confirm that the development of the ENS is not an "all or none" phenomenon.     

Comparison of indapamide and hydrochlorothiazide plus amiloride as a third drug in the treatment of arterial hypertension

Summary In a randomized, double-blind crossover trial, indapamide (IND) 2.5 mg and hydrochlorothiazide 25 mg+amiloride 2.5 mg (HCTZ+a) were found to be equally effective in reducing blood pressure (BP) in 13 patients with moderate to severe hypertension already receiving chronic treatment with a beta blocker and a vasodilatator (supine BP during run-in: 169/103±21/5 mmHg; on IND: 149/91±21/14 mmHg; on HCTZ+A 144/88±23/5 mmHg). Both drugs induced insignificant reductions in body weight, and no change in plasma volume was seen.Serum potassium was significantly reduced on both regimens—the values recorded on IND being significantly lower than those seen on HCTZ+A. Values below 3.0 mmol/l were found in two patients receiving IND, but no subjective side effects were reported. Hyperuricemia occurred with the same frequency on both regimens.It is concluded that IND, just like the thiazide diuretics, is useful as the third drug in patients needing triple drug therapy to control BP, but metabolic adverse effects are not avoided by the choice of this drug.     

Autoimmune thyroiditis: a condition related to a decrease in T-suppressor cells

Monoclonal antibodies recognizing specific antigenic determinants of peripheral T-lymphocytes (OKT3PAN), helper T-cells (OKT4IND) and suppressor T-cells (OKT8SUP) were used to study the immune regulation in autoimmune thyroiditis. An indirect immunofluorescence microscopy method was employed to quantify the number of different subtypes. In addition, B-lymphocytes were also studied using a fluorescent surface Ig detection technique. Patients with autoimmune thyroiditis--independent of their clinical status--show a decrease in the number of suppressor T-lymphocytes. This finding, in agreement with other functional tests, indicates that the autoimmune phenomenon is linked to a decreased T-suppressor activity.     

Notable postnatal alterations in the myenteric plexus of normal human bowel

BACKGROUND: Nitric oxide is the most important transmitter in non-adrenergic non-cholinergic nerves in the human gastrointestinal tract. Impaired nitrergic innervation has been described in Hirschsprung's disease, hypertrophic pyloric stenosis, and intestinal neuronal dysplasia (IND). Recent findings indicate that hyperganglionosis, one of the major criteria of IND, is age dependent. However, information is scanty regarding the neurone density in normal human bowel in the paediatric age group. AIMS: To determine neurone density, morphology, and nitric oxide synthase distribution of the normal myenteric plexus at different ages during infancy and childhood. METHODS: Specimens were obtained from small bowel and colon in 20 children, aged one day to 15 years, at postmortem examination. Whole mount preparations were made of the myenteric plexus, which were subsequently stained using NADPH diaphorase histochemistry (identical to nitric oxide synthase) and cuprolinic blue (a general neuronal marker). The morphology of the myenteric plexus was described and the neurone density estimated. RESULTS: The myenteric plexus meshwork becomes less dense during the first years of life. The density of ganglion cells in the myenteric plexus decreases significantly with age during the first three to four years of life. The NADPH diaphorase positive (nitrergic) subpopulation represents about 34% of all neurones in the myenteric plexus. CONCLUSIONS: The notable decrease in neurone density in the myenteric plexus during the first years of life indicates that development is still an ongoing process in the postnatal enteric nervous system. Applied to the clinical situation, this implies that interpretation of enteric nervous system pathology is dependent on the age of the patient.