Ultrastructural demonstration of abnormal multiaxonal Schwann-cell units in Hirschsprung's disease

The pathophysiology of Hirschsprung's disease is not fully understood. Using light microscopy we have previously demonstrated the absence of a unique Schwann-cell antigen in the circular muscle of aganglionic colon identified by D₇ monoclonal antibody. In an attempt to characterise the morphological changes in neuronal cells at subcellular level, we studied innervation patterns in normal and aganglionic colon by electron microscopy. The most striking observation on ultrastructural serial examination of the entire resected specimen of colon from patients with Hirschsprung's disease was the presence of grossly swollen monoaxonal or oligoaxonal Schwann cell units with loss of cellular contents in the circular muscle of aganglionic colon. The extent of subcellular changes in Schwann cells and axons corresponded with a diminution of immunoreactivity with a panel of neuronal cell antibodies. These ultrastructural findings suggest that degenerative changes in Schwann cells and axons within the circular muscle coat of aganglionic segment may be a significant factor in the pathogenesis of Hirschsprung's disease. Offprint requests to: P. Puri     

Suction biopsy in Hirschsprung's disease.

Seventy two children with symptoms and signs consistent with Hirschsprung''s disease had full thickness and suction rectal biopsies performed. Results were identical with both methods, except for one case of total aganglionosis of the colon. Full thickness biopsy no longer has a place as a screening method.     

Hyperganglionosis mimicking Hirschsprung's disease.

Three patients with hyperganglionosis are reported in whom an initial diagnosis of Hirschsprung's disease was suspected. In one patient there was a classic presentation with constipation, in another Hirschsprung's disease coexisted, and in the third the initial inadequate suction rectal biopsy specimen was suggestive of Hirschsprung's disease on acetylcholinesterase staining. Evidence of hypertrophy and hyperplasia of the intermuscular and submucosal plexuses on a full thickness bowel biopsy specimen was used to confirm the diagnosis of hyperganglionosis, suggested by the characteristic demonstration of moderate increase in the number of acetylcholinesterase stained nerve fibres in the lamina propria mucosae on rectal biopsy. Surgical management was guided by clinical signs. Two patients had colonic resections; the third had temporary stomal diversion. Hyperganglionosis is rarer than Hirschsprung's disease but is known to mimic it. We suggest full thickness bowel specimens are needed to confirm the diagnosis and that inadequate rectal suction biopsies must be interpreted with caution.     

Familial aspects of Hirschsprung's disease.

Twenty-eight of 370 patients (14 families) treated for Hirschsprung's disease (HD) over a 34 year period had a family member with histologically proven HD. These 14 represented 4% of the 351 families: more than one affected child per family in 10 (2.8%) and both parent and child in 2 families. Neuronal intestinal dysplasia (NID) in a parent was associated with total colonic aganglionosis in two siblings of one family which suggests a similar genetic inheritance pattern to HD. Aganglionosis extended beyond the rectosigmoid in 61% of the familial group as opposed to 27% of the non-familial group. A significantly higher number of total colonic aganglionosis (TCA) was noted in those with a family history; 11 out of 28 (39%) as opposed to 19 out of 342 (5.6%) without a family history (p less than 0.001). Fifty percent of males with TCA had a family history but in only 2 cases was this transmitted through a female sibling. Although no significant difference was noted between male and female probands, a three times higher incidence of familial occurrence was noted in females with rectosigmoid disease than in males. Progression of length of segment in succeeding generations was noted in two families. Associated anomalies occurred in 16% without familial occurrence and 11% of the familial group.     

Diagnosis and therapy of ultrashort Hirschsprung's disease.

BACKGROUND: Although ultrashort Hirschsprung's disease (UHD) was enzyme-histochemically characterised about 35 years ago, its existence is still often ignored. The aim of this study is to summarise the clinical diagnostic, incidence, gender ratio, morphological characteristics, and therapy over 15 years. METHODOLOGY: The reliable diagnosis of suspected UHD requires a minimal enema of contrast medium to exclude Hirschsprung's disease (HD). In UHD during pressing or crying no reflux of contrast medium is observed. Final proof of UHD is an enzyme-histochemical biopsy examination of distal rectal mucosa. The biopsies must demonstrate submucosa and be taken from the dentate line and 1 cm, 2 cm, 4 cm and 6 cm above the dentate line. The cryostat sections must be cut 15 microm thick; this thickness is reduced to 4.5 microm by the thawing, spreading and drying of the sections on microscope slides. A reliable diagnosis of UHD needs an enzyme-histochemical acetylcholinesterase reaction of native sections of rectal mucosa. RESULTS: UHD develops with first symptoms of chronic constipation in the second half of the first year of life. The chronic constipation proves to be therapy resistant. In HD constipation occurs in the first weeks of life or after weaning. In contrast to HD, no nerve fibres with increased AChE activity are observed in the lamina propria mucosa. Nets of nerve fibres with increased AChE activity can be found only in the muscularis mucosa and the musculus corrugator cutis ani (MCCA). The therapy of choice has proven to be a partial myectomy of the distal internal sphincter if dilatation of the internal sphincter was ineffective. UHD is either limited to the anal ring, or extends 3 - 4 cm into the distal rectum. Over the past 15 years, UHD had in our series an incidence of 13.4 % of all aganglionoses. The gender ratio of girls to boys was 1 : 2. CONCLUSION: UHD is reliably diagnosed by an AChE reaction in native biopsy sections from the anocutaneous transitional zone and, potentially, from 3 - 4 cm above the pectinate line. As UHD is always accompanied by aganglionosis of the distal internal sphincter, an increase in AChE activity is observed in the nerve fibres of the MCCA. The therapy of choice is a partial myectomy of the distal internal sphincter.     

Enterocolitis in Hirschsprung's disease

During the 5 years 1985–1989, 24 (32%) of 76 patients treated for Hirschsprung's disease (HD) developed enterocolitis, this being present at the time of diagnosis in 10 (13%) infants, 7 of whom were neonates. HD presented as necrotizing enterocolitis in 5 neonates, 4 of whom were premature. The enterocolitis developed postoperatively in 14 (18%) patients, in 7 after an enterostomy and in 7 after a pull-through procedure. Recurrent episodes of enterocolitis occurred in 4 of the patients who developed postoperative enterocolitis. The risk of enterocolitis was increased in girls, in patients with associated Down's syndrome, those with a family history of HD, and those managed by an endorectal pull-through procedure. The Duhamel procedure was associated with a low (5%) incidence of postoperative enterocolitis. Long-segment aganglionosis was not associated with an increased risk of enterocolitis and preoperative enterocolitis conferred no increased risk of postoperative enterocolitis.     

Manometry and histochemistry in the diagnosis of Hirschsprung's disease.

Acetylcholine esterase histochemistry and rectal manometry have been used sequentially to evaluate constipated children and to make the diagnosis of Hirschsprung's disease. When applied together, these modalities give a reliable diagnosis of Hirschsprung's disease, restricting the use of deep rectal biopsy to those patients in whom symptoms are most suggestive of Hirschsprung's disease. Acetylcholine esterase histochemistry may be more reliable than rectal manometry in the newborn and premature periods. It is hoped that the combined use of these techniques may make deep rectal biopsy obsolete except in patients with hypoganglionosis.     

先天性巨结肠与巨结肠同源病的Ret蛋白免疫组织化学研究

目的了解Ret蛋白在先天性巨结肠和具结肠同源病肠组织中的表达,并进一步研究Ret蛋白在先天性巨结肠和巨结肠同源病发病中的作用。方法采用鼠抗人Ret单克隆抗体通过免疫组化SABC法对15例先天性巨结肠,11例巨结肠同源病肠组织Ret蛋白的表达进行研究,10例正常结肠组织作对照。结果Ret蛋白在先天性巨结肠和巨结肠同源病的扩张段,对照组均呈现阳性反应（P〉0．05）,在先天性巨结肠狭窄段大多数表现为阴性反应,极少数出现阳性反应,而巨结肠同源病狭窄段可见Ret免疫反应蛋白阳性细胞,而且偶见巨大的阳性细胞;先天性巨结肠狭窄段分别与巨结肠同源病狭窄段和对照组比较均有统计学意义（P〈0．001）。结论Ret蛋白对先天性巨结肠的发生有重要作用,而与巨结肠同源病的发生无明确关系。     

Prolonged carriage of Clostridium difficile in Hirschsprung's disease.

The role of Clostridium difficile in the aetiology of diarrhoea in children with Hirschsprung's disease was investigated in a prospective longitudinal study. In 64 children with Hirschsprung's disease no significant difference was found in the isolation rate of C difficile in patients with diarrhoea (32%) and without diarrhoea (26%). Comparable isolation rates were found in 47 control children with and without diarrhoea (27% and 16% respectively). The number of strains producing toxin B was similar in the four groups of children. In contrast to the disappearance of C difficile by 12 months of age in the control groups of children, C difficile could be repeatedly isolated from a proportion of children with Hirschsprung's disease over 12 months of age. These findings help to reconcile the existing contradictory reports on the incidence of C difficile in Hirschsprung's disease associated enterocolitis.     

The distributions and coexistence of peptides in nerve fibres of large bowel affected by Hirschsprung's disease

The distributions of nerve fibres immunoreactive for the peptides calcitonin gene-related peptide (CGRP), enkephalin (ENK), neuropeptide Y (NPY), somatostatin (SOM), substance P (SP), and vasoactive intestinal peptide (VIP) and the catecholamine-synthesizing enzyme tyrosine hydroxylase (TH) were studied in healthy colon and samples of ganglionic and aganglionic colon from cases of proven Hirschsprung's disease. Studies of coexistence of reactivities in nerve fibres were performed to predict the possible origins of fibres that are found in the aganglionic bowel, e. g., from sensory or sympathetic ganglia. The muscularis externa of the ganglionic colon contained many nerve fibres immunoreactive for ENK, SP, and VIP, fewer for NPY, and only rare fibres reactive for CGRP, SOM, or TH. In ganglionic colon reactivities for SP and ENK coexisted in nerve fibres in the muscularis externa but in aganglionic colon no ENK immunoreactivity was found and most SP fibres were double-labelled with CGRP reactivity, indicating their probable sensory nature. Abnormally increased numbers of somatostatin-reactive fibres and noradrenergic fibres (marked by TH) were noted in the external muscle, but no coexistence was seen between these reactivities and only a small proportion of the noradrenergic fibres in the muscle showed NPY reactivity although almost all around blood vessels did. Many fibres in the diseased segment had coexistence of NPY and VIP reactivities; these may arise from more orally located intrinsic cell bodies or from pelvic parasympathetic ganglia. In the mucosa of aganglionic colon there was a striking lack of SP-reactive fibres while other fibre types were often normal in number. It is concluded that nerve fibres from sensory ganglia, sympathetic ganglia, nerve cells located more oral in the ganglionated part, and possibly from pelvic parasympathetic ganglia invade the aganglionic bowel in Hirschsprung's disease.     

Diagnosis of Hirschsprung's disease

A prospective study of the accuracy of various diagnostic methods used in the detection of Hirschsprung's disease (syn. congenital intestinal aganglionosis, CIA) in 60 consecutive infants and children was done during the period 1972--76. Every patient underwent a barium enema, a rectal mucosal biopsy, which was prepared for both the demonstration of ganglia and for the assessment of acetylcholinesterase activity (ACE), and anal manometry was performed. In evaluating the clinical history, special emphasis was placed on signs of neonatal ileus. In the group of 10 patients with a definite diagnosis of CIA the results were almost uniform. In the 'non-CIA' group the search for ganglia in biopsy material proved non-confirmatory in nearly half of the cases studied due to the fact that specimens were taken too superficially. The findings pertaining to ACE, barium enema and the results of manometry were at variance or inconclusive of a final diagnosis in 10, 16 and 22% of the performed studies, respectively. The value given to neonatal history proved to be of the same order, i.e., 20% proved to be falsely positive.     

Intestinal atresia and Hirschsprung's disease

Intestinal atresia associated with Hirschsprung's disease has been reported in only 26 cases (20 of small bowel and 6 of colon). Three additional patients are reported, two with associated myelomenigocoele. The significance of the myelomeningocoele and possible aetiological mechanisms of these associations are discussed with particular reference to the role of a vascular accident or the embryological failure of migration of nerve cells. The most likely cause is a volvulus proximal to the aganglionic bowel resulting in the associated atresia. Offprint requests to: S. Cywes     

The electrorectogram in Hirschsprung's disease

The electrorectographic pattern of Hirschsprung's disease (HD) was studied in 14 HD patients with a mean age of 4.6 ± 1.5 years; 7 healthy children acted as controls. Monpolar recordings were made from a silver-silver chloride electrode situated 1 cm from the tip of a 4 F catheter attached to the rectal mucosa by suction. At least four 120-min recording sessions were performed for each subject. No complications were encountered. Regular and reproducible triphasic pacesetter potentials (PP) were recorded from all healthy children, followed randomly by bursts of action potentials (AP). No PP or AP were recorded from patients with HD; the silent electrorectogram (ERG) was reproducible. Since numerous difficulties in histopathologic interpretation affect the reliability of rectal biopsy for the diagnosis of HD, the ERG may play a role in this respect. The ERG is noninvasive and nonradiologic; however, until this investigati tool is substantiated by the work of other investigators, a histologic diagnosis needs to be made before undertaking a pull-through procedure.     

Ondine's curse and Hirschsprung's disease: neurocristopathic syndrome.

We report a female newborn with Ondine's curse and Hirschsprung's disease--neurocristopathic syndrome. The female infant required endotracheal intubation and mechanical ventilation due to apnea which developed soon after birth. She had abdominal distension with bilious vomiting. A barium enema revealed a caliber change at the rectum and rectal biopsies showed no ganglion cells. Colostomy was performed at the age of 17 days. Hypoxemia with hypercapnia was noted during her sleep, and tracheostomy was performed at the age of 55 days. In addition, deafness and pupillary autonomic dysfunction were observed. The definitive surgery for Hirschsprung's disease was performed at the age of 4 months. She is now 2 years old with normal growth but needs ventilator support at home. In this case, we detected no mutation in the RET gene and EDNRB gene.