Intestinal Neuronal Dysplasia Research Articles

Hirschsprung Disease and Intestinal Neuronal Dysplasia Type B: Is There a Difference in the Clinical Presentation of These Two Diseases?

Although the signs and symptoms that comprise the clinical presentation of Hirschsprung disease (HD) and intestinal neuronal dysplasia type B (IND-B) are well established, no studies have specifically compared the clinical characteristics presented by patients with these diseases. We compared the clinical pictures of patients with HD and IND-B at the time of histopathological diagnosis. This was a single-center, retrospective, analytical, and comparative study. We included 119 patients aged 0-15 years diagnosed with HD or IND-B. Information from the medical records of these patients was retrieved to obtain demographic and clinical information at the time of diagnosis. The data were compared statistically according to the characteristics of the variables. Sixty-nine patients (58.0%) were diagnosed with HD, and 50 (42.0%) had IND-B. The HD group had significantly more individuals with symptom onset in the neonatal period (p = 0.001), delayed meconium clearance (p < 0.001), failure to thrive (p = 0.02), and acute complications, such as enterocolitis (p = 0.049) or acute abdominal obstruction (p = 0.031), more commonly requiring emergency surgery (p < 0.001). Patients with IND-B were diagnosed at a significantly older age (p = 0.002). They more commonly had chronic constipation as their main symptom (p = 0.004), with local complications, such as evacuation bleeding (p = 0.007). There were significant differences between the clinical pictures of patients with HD and IND-B. Knowledge of each disease's most common signs and symptoms can help direct diagnostic susception and initial management.

A novel mouse model of intestinal neuronal dysplasia: visualization of the enteric nervous system.

Intestinal neuronal dysplasia (IND) is a congenital anomaly affecting gastrointestinal neural innervation, but the pathogenesis remains unclear. The homozygous Ncx/Hox11L.1 knockout (Ncx-/-) mice exhibit megacolon and enteric ganglia anomalies, resembling IND phenotypes. Sox10-Venus transgenic mouse were used to visualize enteric neural crest cells in real time. This study aims to establish a novel mouse model of Sox10-Venus+/Ncx-/- mouse to study the pathogenesis of IND. Sox10-Venus+/Ncx-/- (Ncx-/-) (n = 8) mice and Sox10-Venus+/Ncx+/+ controls (control) (n = 8) were euthanized at 4-5weeks old, and excised intestines were examined with fluorescence microscopy. Immunohistochemistry was performed on tissue sections with neural marker Tuj1. Ncx-/- mice exhibited dilated cecum and small intestine. Body weight of Ncx-/- mice was lower with higher ratio of small intestine length relative to body weight. The neural network (Sox10-Venus) was observed along the intestine wall in Ncx-/- and control mice without staining. Ectopic and increased expression of Tuj1 was observed in both small intestine and proximal colon of Ncx-/- mice. This study has established a reliable animal model that exhibits characteristics similar to patients with IND. This novel mouse model can allow the easy visualization of ENS in a time- and cost-effective way to study the pathogenesis of IND.

Retracted: Comparison of Outcomes between Two Surgical Techniques for Patients with Intestinal Neuronal Dysplasia.

[This retracts the article DOI: 10.1155/2022/9165651.].

Association between Clinical and Histopathological Findings in Intestinal Neuronal Dysplasia Type B: An Advance towards Its Definition as a Disease.

Intestinal neuronal dysplasia type B (IND-B) is a controversial entity that affects the submucosal nerve plexus of the distal intestine. The lack of definition of the causal relationship between histological findings and clinical symptoms has been identified as the primary point to be elucidated in the scientific investigation related to IND-B, which is essential for it to be considered a disease. To investigate the relationship between histopathological findings and symptoms in a series of patients with IND-B. Twenty-seven patients with histopathological diagnosis of IND-B, according to the Frankfurt Consensus (1990), who underwent surgical treatment through colorectal resections were included. Data from medical records regarding the clinical picture of the patients at the time of diagnosis, including the intestinal symptom index (ISI) and a detailed histopathological analysis of the rectal specimens, were retrieved. Exploratory factor analysis was performed, applying the principal components method for clusters with Varimax rotation. Two factors were determined: the first, determined by histopathological and clinical variables, and the second, composed of the main symptoms presented in patients with IND-B, including ISI. Factorial rotation showed the association between the two factors and, through a graph, demonstrated the proximity between ISI values and histopathological alterations. There was evidence of an association between the clinical features presented by patients with IND-B and the histopathological findings of the rectal samples. These results support the understanding of IND-B as a disease.

Intestinal neuronal dysplasia type B of the duodenum in an adolescent patient

Intestinal neuronal dysplasia type B (IND-B) is a rare neuromuscular, dysmotility disorder that typically affects the lower gastrointestinal tract, commonly presenting as chronic constipation in children. We present the first pediatric case report of successful surgical management of IND-B affecting the duodenum as a likely chronic adaptive sequela to prior obstruction from superior mesenteric artery syndrome.

Sporadic diffuse gastric ganglioneuromatosis causing gastric outlet obstruction in an adult patient: A rare case report and literature review

: Gastrointestinal ganglioneuromatosis is a rare benign neural neoplasm, consists of hyperplasia of nerve fibers, ganglion cells, and schwann cells in the wall of stomach and intestine. The disease may affect any part of the gastrointestinal tract but involvement of stomach is extremely unusual. Mainly are of three types: polypoid ganglioneuromas, ganglioneuromatous polyposis, and diffuse ganglioneuromatosis. To our knowledge, only two cases were reported as diffuse ganglioneuromatosis located in the stomach.: 27-years male patient, admitted with complaint of dysphagia for both solid and liquid and recurrent non-bilious vomiting since 1 week underwent Billroth - 1 procedure and antral segment of stomach was sent for histopathological examination. A histopathological diagnosis of Diffuse Gastric Ganglioneuromatosis with muscular mural hypertrophy was made. The presence of ganglion cells and positive staining for S-100 and NSE confirms the diagnosis.Obstructive gastrointestinal symptoms with a possibility of diffuse ganglioneuromatosis is mainly a disease of young children and infant. But an adult patient, which is an unusual age group, presenting with obstructive gastrointestinal symptoms, the rare possibility of Diffuse ganglioneuromatosis and intestinal neuronal dysplasia should be considered and investigated in order to avoid unnecessary and extensive surgical intervention.

Intestinal neuronal dysplasia presenting as psoas abscess: A case report

Intestinal neuronal dysplasia (IND) is a rare condition mainly affecting the children. Constipation and abdominal distension have been reported as common manifestations. In addition, the reports about adult cases are scarce. A 31-year-old man presented with pain in his left hip and intermittent fever for 1 month. The whole abdomen CT and pelvic contrast-enhanced MRI revealed a left psoas abscess (PA). The patient has been given anti-infective treatment and underwent CT-guided drainage of left PA with a temporary drain. But the patient's condition did not improve significantly. Then, the colonoscopy revealed that it may be the PA secondary to inflammatory bowel disease. But the pathology was not in line with inflammatory bowel disease. We finally performed an ileostomy surgery and took the whole layer of intestinal wall for biopsy. The pathological result revealed that a large number of proliferative ganglion cells and circuitous hyperplastic nerve fibers were found in the submucosa and muscular layer of the intestinal wall. Given pathological results and clinical manifestations, the patient was diagnosed with IND-B. In this case, we first report an extremely rare case of adult IND manifesting as PA. So, this unusual case provides a new supplement to adult cases of IND.

Diagnostic utility of Bcl-2 immunohistochemical expression in pediatric functional bowel obstruction cases with ganglionated specimens

BackgroundFunctional disturbances of the gastrointestinal tract are caused by a number of neurodysplastic conditions, including diseases that are rarer than Hirschsprung’s disease (HSCR), such as ganglion cell immaturity and intestinal neuronal dysplasia (IND). Bcl-2 shows positive immunoreactivity in degenerative and immature ganglion cells (IGCs). This work evaluates the implication of the immunohistochemical expression of Bcl-2 in cases of pediatric intestinal pseudo-obstruction (IPO) with ganglionated specimens.This 2-year prospective observational study was conducted from January 2017 to December 2019 on all intestinal specimens for cases referred from a pediatric surgery department with clinical data suggesting IPO (abdominal distension > 3 months, intermittent bilious vomiting, no radiological evidence of the transitional zone of HSCR or evidence of mechanical obstruction). The exclusion criteria were patients with IPO related to myopathic disorders, aganglionic biopsies, and specimens with inadequate tissue blocks. The same number of intestinal specimens of cases without IPO was used as a control group. All specimens underwent a histologic examination of ganglion cells and nerve bundles and were also evaluated for Bcl-2 immunohistochemical expression.ResultsTwenty-one specimens were analyzed, including six colonic resection specimens and 15 intestinal biopsies taken by full-thickness transanal biopsy (n = 12) or incisional biopsy from inadequately functioning stomas (n = 3). The mean ± standard deviation age of the patient cohort was 22 ± 7.4 months (range, 19–153 months), and there were 13 (61.9%) male patients. Bcl-2 protein was strongly positive in 57.1% of the cases, weakly positive in 19%, and negative in 23.8% as well as in all 21 control specimens. A highly significant correlation was observed between Bcl-2 expression and ganglion cell number (P < 0.001), where all hyperganglionic specimens showed strong positivity compared with the hypoganglionic and adequately ganglionic specimens. A positive association was also found between Bcl-2 expression and IND compared with other disorders of dysganglionosis (P = 0.04) and post-HSCR cases (P = 0.002).ConclusionBcl-2 immunohistochemistry is a valuable tool to diagnose allied disorders of HSCR through its expression in IGCs, which are difficult to identify by conventional hematoxylin and eosin staining.

S3358 Exploring Probe-Based Confocal Laser Endomicroscopy as a Novel Tool in Graft Monitoring After Small Bowel Transplantation

Introduction: The most common and severe complication early in Small Bowel Transplant (SBT) is graft rejection making timely detection key to graft preservation. Endoscopic surveillance with protocol biopsy is the gold standard for graft monitoring. Confocal Laser Endomicroscopy (CLE) has emerged as a modality for real-time diagnosis at a histological scale. However, its role in SBT is not known. Herein, we report 3 cases of SBT patients and compare CLE results with standard protocols. Case Description/Methods: Case 1: A 73-year-old man underwent SBT for sclerosing mesenteritis and total enterectomy. CLE and ileoscopy was performed 15 times post-transplant without findings of acute cellular rejection (ACR). Patient expired of sepsis on day 30 post-transplant. Case 2: A 17-year-old male with familial adenomatous polyposis complicated by total colectomy, hepatoblastoma s/p resection and chemotherapy underwent SBT for large desmoid tumor requiring total enterectomy. CLE and ileoscopy was performed 14 times post-transplant without findings of ACR. Case 3: A 23-year-old female underwent SBT for intestinal neuronal dysplasia after chronic rejection from 1st transplant 10 years ago. Ileoscopy done at 20 days post-transplant showed congested and ulcerated mucosa in the proximal ileum. CLE images and histopathology showed findings consistent with ACR. Immunosuppression was increased and reassessment with CLE and ileoscopy done. Discussion: Cases 1 and 2 show normal CLE assessment. Normal villus architecture, non-edematous, and adequate microcirculation suggesting low suspicion for ACR. This correlated with normal pathology characterized by less than 6 apoptotic bodies per 10 crypts, no intravascular neutrophils, and negative viral cytopathology. However, in case 3, CLE images showed significant villus congestion and decreased microcirculation (Figure) indicative of ACR. Biopsies confirmed ACR. Rejection protocols were rapidly initiated.Repeat CLE and ileoscopy showed recovery of mucosal congestion and return of microcirculation. This correlated with complete resolution of ACR on pathology. In the absence of endoscopic criteria for small bowel graft rejection, biopsies are the gold standard. Enhanced technology for in vivo visualization of grafted bowel is needed and would allow for examination of a greater surface area of the bowel than limited biopsies. CLE is a promising tool that could significantly improve early transplant graft monitoring. Further data and prospective studies are needed.Figure 1.: CLE images showing significant villus congestion, and decreased microcirculation.

Comparison of Outcomes between Two Surgical Techniques for Patients with Intestinal Neuronal Dysplasia.

Objective Hirschsprung disease (HSCR) is a serious congenital intestinal disease with a prevalence of 1/5000. HSCR remains one of the most severe congenital malformations of the abdominal organs in children that require complex reconstructive surgery. This study is aimed at investigating the clinical analysis of ileal Santulli stoma and ileal double-lumen stoma in children diagnosed with intestinal neuronal dysplasia (IND). Methods Retrospective analysis was performed on the children who were admitted to our hospital for intestinal obstruction from January 2014 to January 2019, underwent fistula operation and fistula closure operation, and were diagnosed with IND. According to the different modes of fistula, the children were divided into ileal Santulli stoma group and ileal double-lumen stoma group. The body weight of the children in the two groups during the second stage of fistula closure operation was compared. The number of hospitalizations due to enteritis and dehydration during the two operations was compared. Results A total of 23 cases (12 males and 11 females) were included in this study, including 10 cases in the Santulli group and 13 cases in the ileal double-lumen stoma group. There were no significant differences in baseline data and fistula location between the two groups. Compared with the ileal double-lumen stoma group, the Santulli stoma group had significantly higher weight of fistula precursor (P < 0.05), the interval between two operations was shorter (P < 0.05), there is less hospitalization for enteritis and dehydration during the two operations (P < 0.05), and there is less economic cost after fistula (P < 0.05). Conclusion The clinical effect of ileum Santulli fistula is significantly better than double-lumen ileum fistula, which is not only beneficial to the growth and development of children after the first fistula but also can shorten the time of fistula closure, reduce the incidence of dehydration, and reduce the economic burden of family members. Therefore, it is worthy of clinical promotion and application.

PTEN Immunohistochemistry.

Intestinal neuronal dysplasia type B (IND B) is a complex entity involving the enteric nervous system, clinically manifested with constipation in infancy. Diagnosis has been established by histopathologic analysis of rectal biopsies. However, the criteria for the diagnosis have been questioned and modified, hindering diagnostic practice. To analyze the applicability of PTEN immunohistochemistry in the diagnosis of IND B and to compare with control cases and cases of Hirschsprung disease (HD). PTEN immunohistochemical expression was analyzed in colorectal samples from 29 cases of IND B and compared with 4 control cases and 6 cases of HD. The pattern of PTEN immunoexpression was analyzed in glial cells of the submucosal and myenteric nerve plexuses and in neural fibrils of the muscularis propria using a scoring system. Marked reduction or absence of PTEN expression was observed in glial cells of the submucosal nerve plexuses in all cases of the IND B group and in the myenteric nerve plexuses in 28 of 29 cases (96.5%). Lack of PTEN expression was detected in neural fibrils within the muscularis propria in 21 of 29 cases (72%) of the IND B group. PTEN expression was positive in the same neural structures of the control and HD groups. PTEN immunohistochemistry may be a valuable tool in the diagnostic evaluation of IND B. Lack of or reduction of PTEN expression in neural fibrils within the muscularis propria suggests that involvement of the neuromuscular junction may be a key event in the pathogenesis of the motility disturbance occurring in IND B.

Role of GDNF, GFRα1 and GFAP in a Bifidobacterium-Intervention Induced Mouse Model of Intestinal Neuronal Dysplasia.

ObjectiveTo investigate the effects of glial cell-derived neurotrophic factor (GDNF), GDNF family receptor alpha 1 (GFRα1), and glial fibrillary acidic protein (GFAP) on colonic motility in a mouse model of intestinal neuronal dysplasia by intervention with Bifidobacterium and to explore the influence of Bifidobacterium on enteric glial cells (EGCs).MethodsWestern blotting and qRT-PCR were employed to detect the expression of GFRα1 and GFAP in colonic tissues of mice with or without Tlx2 mutations, and ELISA was used to detect the expression of GDNF in serum. IHC was used to detect the appearance of the ganglion cells. Subsequently, Tlx2 homozygous mutant (Tlx2−/−) mice were treated with Bifidobacterium. Colonic motility was measured before and after intervention by measuring the glass bead expelling time. The variations in abdominal circumference and GDNF, GFRα1, and GFAP expression were measured. In addition, 16SrRNA gene sequencing was performed to detect the abundance of the intestinal microbiota.ResultsThe mRNA and protein expression of GFRα1 and GFAP was decreased in the colonic tissues of Tlx2−/− mice and GDNF expression was decreased in serum compared with Tlx2+/− and WT mice. After confirming the colonization of Bifidobacterium by 16S rRNA gene sequencing, the expelling time and abdominal distension were ameliorated, and the expression of GFAP, GDNF, and GFRα1 was increased.ConclusionsThe expression of GDNF, GFRα1, and GFAP is associated with colonic motility. The altered expression of EGC-related factors suggested that Bifidobacterium may be involved in the EGC activation process. The amelioration of IND symptoms after intervention with Bifidobacterium prompted the elicitation of adjuvant therapy.

A successful treatment of total colonic aganglionosis in a child

Introduction. The earliest mention of total colonic aganglionosis, as the form of Hirschsprung’s disease, is dated 1926. According to various sources, total colonic aganglionosis is met approximately in 1 per 500 000 live births. Currently, the curative strategy in total colonic aganglionosis is strongly personalized. There are various surgical techniques for managing this severe congenital colon anomaly. Recent achievements in pediatric coloproctology have minimized the mortality rate in children with intestinal neuronal dysplasia and total colonic aganglionosis. However, despite of the evident progress in coloproctology, questions about the volume of reconstructive surgery in total colonic aganglionosis, application of “reservoir” surgical techniques so as to promote child’s further development and socialization are still unsolved.Material and methods. The patient was successfully treated with a reservoir technique which included an ileo-transverse anastomosis by Martin-Kimur with the formed conduit pulled to the anterior abdominal wall. Then, a radical intervention was made which consisted of combined abdominoperineal transanal endorectal pull through (TEPT) of the ileo-transverse conduit/reservoir, of the resection of rudimentary colon and a coloanal anastomosis by Boley.Conclusion. The obtained small intestine-colon reservoir in total aganglionosis allows to preserve motor function due to peristaltic movements of the small intestine wall and to prevent intestinal obstruction, thereby facilitating the physiological function of the intestine. In addition, the colon wall in the reservoir saves the area of absorbing surface and minimizes water-electrolyte disorders.

Challenges in the diagnosis of intestinal neuronal dysplasia type B: A look beyond the number of ganglion cells.

Intestinal neuronal dysplasia type B (IND-B) is a controversial condition among gastrointestinal neuromuscular disorders. Constipation is its most common clinical manifestation in patients. Despite intense scientific research, there are still knowledge gaps regarding the diagnostic criteria for IND-B in the histopathological analysis of rectal biopsies. The guidelines published in the past three decades have directed diagnostic criteria for quantifying the number of ganglion cells in the nervous plexus of the enteric nervous system. However, it is very complex to distinguish numerically what is pathological from what is normal, mainly because of the difficulty in determining a reliable control group composed of healthy children without intestinal symptoms. Thus, a series of immunohistochemical markers have been proposed to assist in the histopathological analysis of the enteric nervous system. Several of these markers facilitate the identification of other structures of the enteric nervous system, in addition to ganglion cells. These structures may be related to the etiopathogenesis of IND-B and represent new possibilities for the histopathological diagnosis of this disease, providing a view beyond the number of ganglion cells. This review critically discusses the aspects related to the disease definitions and diagnostic criteria of this organic cause of constipation.

The Role of Bcl-2 in Pediatric Functional Bowel Obstruction Cases with Ganglionated Specimens

Abstract Background Functional disturbances of the gastrointestinal tract is caused by a number of neurodysplastic conditions including other diseases, rarer than the Hirschsprung’s disease dysganglionosis, such as ganglion cells immaturity including Intestinal Neuronal Dysplasia (IND). Bcl-2 shows positive immunoreactivity in the degenerative and immature ganglion cells . Aim This work evaluates the role of immunohistochemical expression of Bcl-2 in pediatric functional bowel obstruction cases with ganglionated specimens. Material and Methods Twenty one cases of functional bowel obstruction were enrolled in our study including the Hirschsprung Allied Disorders (HAD) and operated cases of Hirschsprung’s Disease (HD) with adequate ganglionated resection margins and presented by obstructive symptoms following surgery. All cases underwent histologic examination of ganglion cells and nerve bundles as well as evaluation for Bcl-2 immunohistochemical expression. Results 57.1% of the cases were strongly positive for Bcl-2 protein, 19% were weakly positive and 23.8 % were negative. A highly significant correlation was demonstrated between Bcl-2 expression and ganglion cell number ( P &amp;lt; 0.001 ) where all hyperganglionic specimens showed strong positivity in contrast to the hypoganglionic and adequately ganglionic ones. A positive association was also found between Bcl-2 expression and IND in comparison to other disorders of dysganglionsis (P = 0.04) and recurrent HD cases (P = 0.002). A significant correlation was also detected between ganglion cell number and nerve bundle hypertrophy (P = 0.02). Conclusion Bcl-2 immunohistochemistry can be used as a valuable tool in the diagnosis of Hirschsprung’s disease allied disorders through its expression in the immature ganglion cells which are difficult to be identified by conventional H &amp; E staining.

Enteric nervous system with chronic mesenteric ischemia: experimental study

Introduction. The pathology of the enteric ganglia can lead to different diseases (Hirschsprungs, neuronal intestinal dysplasia, ganglioneuromatosis, and Chagasse). Causes of acquired dysganglionosis remained unclear. Some authors hypothesized that pathology of the enteral nervous system may be secondary to intestinal ischemia.&#x0D; Aim. To investigate the intestinal function and histological changes of the colon in rats with chronic ischemia.&#x0D; Materials and methods. A total of 20 Sprague Dawley rats underwent surgery (ligation of the terminal mesenteric vessels next to the descending colon). The appetite of animals were checked, and stool were collected after the procedure. Reoperation was performed after 7 (n = 1), 9 (n = 2), 12 (n = 2), 14 (n = 1), 21 (n = 1), 42 (n = 1), 53 (n = 1), and 62 (n = 1) days. The diameter of the colon and changes of the serosa were visualized. In the experimental group, two samples biopsy was performed (ischemic and normal colon).&#x0D; Results. Functional changes were observed in 90% of rats after the ligation of mesenteric vessels (constipation/impact, softening stool/diarrhea, and hemocolitis). Colonic stenosis of the ischemic area in 30% was detected. 70% animals have the intestinal dilatation above the ischemic segment (partial bowel obstruction). Necrosis of the ischemic colon was observed in 20%. Spontaneous fixation of the omentum to the ischemic segment was found in 40% animals. A microscopically inflamed infiltration of the mucosa in the ischemic zone (70%) and in normal colon (50%) was revealed in the ligation group. The number of the enteric ganglia decreased in the ischemic segment.&#x0D; Conclusion. Functional disorders (colitis and obstruction) and morphological changes (inflammation and ganglion cells pathology) were found in rats with chronic mesenteric ischemia.

What should be the treatment for intestinal neuronal dysplasia type B? A comparative long-term follow-up study

PurposeTo present the long-term follow-up outcomes of patients with intestinal neuronal dysplasia type B (IND-B) managed either conservatively or surgically. MethodsWe conducted an ambispective, observational, longitudinal, and comparative study. Clinical data were reviewed at the start of treatment. After a minimum period of five years, the patients participated in semi-structured interviews in which the bowel function score (BFS) was applied to assess intestinal function, a proposed intestinal symptom index (ISI) to assess clinical symptoms, and a classification of clinical prognosis to assess treatment success. Comparisons between the two types of treatment were performed by evaluating pre- and post-treatment criteria. ResultsFifty patients diagnosed with IND-B were included in the study. Thirty-eight patients underwent surgical treatment (26 elective surgical treatment for primary colorectal resection and 12 emergency colostomies for intestinal obstruction or enterocolitis). Twelve patients were managed conservatively. With the exception of the patients who required an emergency operation (n = 12), the two groups were composed of patients with severe constipation who had similar clinical and functional characteristics at the time of IND-B diagnosis. A better clinical response was observed in patients submitted to conservative treatment, with a greater increase in the BFS (16.5 [-4/+18] versus 4 [-15/+17]; p = 0.001), indicating better bowel function and a more pronounced drop in ISI (-6 [-7/-4] versus -4 [-6/+1]; p = 0.015), suggesting fewer symptoms. The percentage of patients who had a successful treatment was higher in the group treated conservatively (72.7% versus 42.3%; p = 0.03). ConclusionConservative management showed better long-term outcomes than surgical management in children with IND-B.

Effect of Neuroligin1 and Neurexin1 on the Colonic Motility in a Mouse Model of Neuronal Intestinal Dysplasia.

Aim To investigate the expressions of neuroligin1 (NL1) and neurexin1 (NX1) in a mouse model of neuronal intestinal dysplasia (Tlx2−/− mice) and to explore their effects on colonic motility. Methods Immunohistochemistry staining was employed to explore the histological appearances of NL1, NX1, the presynaptic marker of glutamatergic synapses VGLUT1, and the subunit of NMDA receptors of NR1 in the colon of mice with or without Tlx2 mutation. Western blotting and qRT-PCR were performed to detect their relative expressions in the colon. Colonic motility was measured by a glass bead technique. Then, the Tlx2−/− mice were intervened by Huperzine A. Variations on expressions of NL1, NX1, VGLUT1, and NR1 and variations on colonic motility were measured. Additionally, serum concentrations of Glu were measured by ELISA. Results Immunohistochemistry staining reveals that NL1, NX1, VGLUT1, and NR1 were mainly concentrated in the myenteric plexus of ENS. Compared to those in WT and Tlx2+/- mice, expressions of NL1 and NX1 in colon of Tlx2−/− mice were upregulated with increased VGLUT1 and NR1 abundances and impaired colonic motility (P < 0.05). After intervention, the upregulated expressions of NL1 and NX1 were decreased with a correlated reduce of VGLUT1 and NR1 and a recovery of the impaired colonic motility (P < 0.05). After intervention, the upregulated expressions of NL1 and NX1 were decreased with a correlated reduce of VGLUT1 and NR1 and a recovery of the impaired colonic motility (P < 0.05). After intervention, the upregulated expressions of NL1 and NX1 were decreased with a correlated reduce of VGLUT1 and NR1 and a recovery of the impaired colonic motility (Conclusion NL1 and NX1 are closely related to the colonic motility through their effects of targeting the formation of glutamatergic synapses and may be involved in the pathogenesis of NID. The variations of serum Glu seem to be a potential and less painful auxiliary measure for colonic motility and NID.

Efficacy of Sox10 Promoter Methylation in the Diagnosis of Intestinal Neuronal Dysplasia From the Peripheral Blood

OBJECTIVES:Intestinal neuronal dysplasia (IND) is a common malformation of the enteric nervous system. Diagnosis requires a full-thickness colonic specimen and an experienced pathologist, emphasizing the need for noninvasive analytical methods. Recently, the methylation level of the Sox10 promoter has been found to be critical for enteric nervous system development. However, whether it can be used for diagnostic purposes in IND is unclear.METHODS:Blood and colon specimens were collected from 32 patients with IND, 60 patients with Hirschsprung disease (HD), and 60 controls. Sox10 promoter methylation in the blood and the Sox10 expression level in the colon were determined, and their correlation was analyzed. The diagnostic efficacy of blood Sox10 promoter methylation was analyzed by receiver operating characteristic curve.RESULTS:The blood level of Sox10 promoter methylation at the 32nd locus was 100% (90%–100%; 95% confidence interval [CI], 92.29%–96.37%) in control, 90% (80%–90%; 95% CI, 82.84%–87.83%) in HD, and 60% (50%–80%; 95% CI, 57.12%–69.76%) in IND specimens. Sox10 promoter methylation in the peripheral blood was negatively correlated with Sox10 expression in the colon, which was low in control, moderate in HD, and high in IND specimens (r = −0.89). The area under the curve of Sox10 promoter methylation in the diagnosis of IND was 0.94 (95% CI, 0.874–1.000, P = 0.000), with a cutoff value of 85% (sensitivity, 90.6%; specificity, 95.0%). By applying a cutoff value of 65%, promoter methylation was more indicative of IND than HD.DISCUSSION:The analysis of Sox10 promoter methylation in the peripheral blood can be used as a noninvasive method for IND diagnosis.

Identification of potential molecular pathogenesis mechanisms modulated by microRNAs in patients with Intestinal Neuronal Dysplasia type B

This study proposed to determine global microRNA (miRNA) expression and miRNA-regulated pathways in Intestinal Neuronal Dysplasia type B (IND-B). Fifty patients (0–15 years old) with IND-B were included in the study. Peripheral blood samples were collected from all 50 patients and from 10 healthy asymptomatic children (controls). Rectal biopsies were collected from 29/50 patients; biopsy tissues were needle microdissected to isolate the different intestinal layers, for molecular analysis. Global miRNA expression was determined using TaqMan arrays. Correlation analysis between miRNA expression in plasma and biopsy samples as well as among tissues derived from the distinct intestinal layers was performed. Computational approaches were used for miRNA target prediction/identification of miRNA-regulated genes and enriched pathways biologically relevant to IND-B pathogenesis. miRNAs were statistically significantly deregulated (FC ≥ 2 and p ≤ 0.05) in submucosal and muscular layers: over-expressed (miR-146a and miR-146b) and under-expressed (miR-99a, miR-100, miR-130a, miR-133b, miR-145, miR-365, miR-374-5p, miR-451). Notably, let-7a-5p was highly over-expressed in patient plasma compared to healthy controls (FC = 17.4). In addition, miR-451 was significantly under-expressed in both plasma and all biopsy tissues from the same patients. Enriched pathways (p < 0.01) were axon guidance, nerve growth factor signalling, NCAM signalling for neurite out-growth, neuronal system and apoptosis. miRNA expression is deregulated in the submucosa and muscular layers of the rectum and detected in plasma from patients with IND-B. Biologically enriched pathways regulated by the identified miRNAs may play a role in IND-B disease pathogenesis, due to the activity related to the neurons of the enteric nervous system.