Identification of signaling pathways that specify a subset of migrating enteric neural crest cells at the wavefront in mouse embryos.

During enteric nervous system (ENS) development, pioneering wavefront enteric neural crest cells (ENCCs) initiate gut colonization. However, the molecular mechanisms guiding their specification and niche interaction are not fully understood. We used single-cell RNA sequencing and spatial transcriptomics to map the spatiotemporal dynamics and molecular landscape of wavefront ENCCs in mouse embryos. Our analysis shows a progressive decline in wavefront ENCC potency during migration and identifies transcription factors governing their specification and differentiation. We further delineate key signaling pathways (ephrin-Eph, Wnt-Frizzled, and Sema3a-Nrp1) utilized by wavefront ENCCs to interact with their surrounding cells. Disruptions in these pathways are observed in human Hirschsprung's disease gut tissue, linking them to ENS malformations. Additionally, we observed region-specific and cell-type-specific transcriptional changes in surrounding gut tissues upon wavefront ENCC arrival, suggesting their role in shaping the gut microenvironment. This work offers a roadmap of ENS development, with implications for understanding ENS disorders.

Mesenchymal Stem Cells Attenuates Hirschsprung Disease-Associated Enterocolitis by Reducing M1 Macrophages Infiltration via COX-2 Dependewhant Mechanism.

OBJECTIVE AND DESIGN: Hirschsprung disease-associated enterocolitis (HAEC) is a common life-threatening complication of Hirschsprung disease (HSCR). We aimed to investigate the effectiveness, long-term safety and the underlying mechanisms of Mesenchymal stem cells (MSCs) based therapy for HAEC. MATERIAL OR SUBJECTS: Specimens from HSCR and HAEC patients were used to assess the inflammatory condition. Ednrb knock-out mice was used as HAEC model. MSCs was intraperitoneally transplanted into HAEC mice. The therapy effects, long-term outcome, safety and toxicity and the mechanism of MSCs on the treatment of HAEC were explored in vivo and in vitro. RESULTS: Intestinal M1 macrophages infiltration and severe inflammation condition were observed in HAEC. After the injection of MSCs, HAEC mice showed significant amelioration of the inflammatory injury and inhibition of M1 macrophages infiltration. The expression levels of pro-inflammatory cytokines (TNF-α and IFN-γ) were decreased and anti-inflammatory cytokines (IL-10 and TGF-ß) were increased. In addition, we found that effective MSCs homing to the inflamed colon tissue occurred without long-term toxicity response. However, COX-2 inhibitor could diminish the therapeutic effects of MSCs. Using MSCs and macrophages co-culture system, we identified that MSCs could alleviate HAEC by inhibiting M1 macrophages activation through COX-2-dependent MAPK/ERK signaling pathway. CONCLUSIONS: MSCs ameliorate HAEC by reducing M1 macrophages polarization via COX-2 mediated MAPK/ERK signaling pathway, thus providing novel insights and potentially promising strategy for the treatment or prevention of HAEC.

Identification and validation of the common pathogenesis and hub biomarkers in Hirschsprung disease complicated with Crohn's disease.

Background: Although increasing evidence has supported that Hirschsprung disease (HSCR) is the risk factor for children developing Crohn's disease (CD), the common mechanism of its co-occurrence remains unknown. The purpose of this study is to further explore the underlying mechanism and biomarkers for the co-occurrence of HSCR and CD. Methods: The Gene Expression Omnibus (GEO) database was used to obtain gene expression profiles for CD (GSE95095) and HSCR (GSE98502). Following the identification of the shared differentially expressed genes (DEGs) of CD and HSCR, functional annotation, protein-protein interaction (PPI) network creation, and module assembly were performed to discover hub genes. RT-qPCR was performed to validate the expression of the hub genes in HSCR samples. The receiver operating characteristic (ROC) curve was utilized to assess the accuracy of the hub genes as biomarkers in predicting CD in both the training dataset and test dataset. Results: A total of 103 common DEGs (50 downregulated genes and 53 upregulated genes) were chosen for further investigation. The importance of chemokines and cytokines in these two disorders is highlighted by functional analysis. MCODE plug identified three important modules, which functionally enriched the immune system process. Finally, nine hub genes were identified using cytoHubba, including IL1B, IL10, CXCL10, ICAM1, EGR1, FCGR3A, S100A12, S100A9, and FPR1. The nine hub genes were mainly enriched in immune- and inflammation-related pathways. External data profiles and RT-qPCR confirmed the expression of the nine hub genes in HSCR and CD. ROC analysis revealed that the nine hub genes had a strong diagnostic value. Conclusion: Our study reveals the common pathogenesis of HSCR and CD. These hub genes and diagnostic models may provide novel insight for the diagnosis and treatment of HSCR complicated with CD.

Transanal full-thickness pull-through approach in the treatment of anastomotic leakage after operation for Hirschsprung disease.

BACKGROUND: Anastomotic leakage (AL) is one of the most perplexing complications that can occur following a radical operation to treat Hirschsprung disease (HSCR). The purpose of this study was to document our experience with anastomotic leakage following radical HSCR surgery to enhance therapeutic effect and prognosis. METHODS: Between January 2007 and April 2021, a retrospective study was conducted on 12 children who developed anastomotic leakage following radical surgery for HSCR. Medical records were analyzed to determine the clinical manifestations, primary surgical procedures, evaluation methods, surgical plans, and outcomes of the patients. To assess postoperative bowel function, the Rintala score was used. RESULTS: The Soave procedure was used as the primary surgical method in seven cases (58.3%), the Swenson procedure was used in four cases (33.3%), and the Rehbein procedure was used in one case (8.3%). Enterostomy (10, 83.3%) and conservative treatment (2, 16.7%) were performed when anastomotic leakage was diagnosed. Two patients who directly closed stoma without redoing pull-through both accepted enterostomy within 48 h. One female with anastomotic fistula who was closed leakage or fistula in situ had to endure lifelong stoma. Other patients who underwent redo pull-through procedures had normal bowel function. Seven patients underwent a redo pull-through procedure. Three of them preferred the transanal full-thickness pull-through (FTPT) approach, while four preferred the Soave technique. Three children had mild postoperative soiling, which improved with conservative treatment. Bowel function score was 17.5 ± 1.1. CONCLUSION: Enterostomy should be performed immediately if anastomotic leakage occurs. After leakage, it is necessary to redo the pull-through procedure in an anastomotic fistula or anastomotic stricture. Transanal FTPT reconstruction is an effective method for repairing anastomoses and leakage.

Long-term outcomes of single-incision laparoscopic technique in Soave procedure compared with heart-shaped anastomosis for Hirschsprung disease.

PURPOSE: This retrospective study compared the long-term outcomes of single-incision laparoscopy-assisted Soave procedure (SILSP) with single-incision laparoscopy-assisted heart-shaped anastomosis (SILHSA) in patients with Hirschsprung disease (HSCR). METHODS: Patients diagnosed with HSCR that underwent SILSP or SILHSA between January 2009 and January 2015 at our institute were enrolled in this retrospective study. Data on the clinical characteristics, perioperative complications, and postoperative quality of life were retrospectively collected and analyzed. RESULTS: There were 109 patients in the SILSP group and 95 patients in the SILHSA group. No differences in clinical characteristics, including age, weight, hospitalization length, blood loss volume, and operation time, were noted between the two groups. The incidence rates of constipation and soiling were lower in the SILHSA group than those in the SILSP group. The SILHSA group showed lower scores in constipation and soiling compared with the SILSP group, indicating a better surgical outcome for patients receiving SILHSA procedure. CONCLUSION: SILHSA is a feasible and reliable minimally invasive surgical procedure for patients with HSCR. Patients who underwent SILHSA had lower incidence rates of constipation and soiling than patients who underwent SILSP, suggesting that SILHSA could be a better choice for patients with HSCR.

A Study of Three-Dimensional Versus Two-Dimensional Laparoscopic Surgery in Resection of Congenital Choledochal Cyst of Children and Jejunum Roux-en-Y Anastomosis.

Background: This study aimed to explore the clinical efficacy of three-dimensional (3D) laparoscopic surgery in the treatment of congenital choledochal cysts (CCCs) by comparing it with two-dimensional (2D) laparoscopic surgery. Patients and Methods: We retrospectively reviewed data of 155 pediatric patients who underwent surgical treatment of choledochal cysts between January 2014 and December 2017. We divided the patients into two groups according to the surgical method used-a 3D laparoscopic group (N = 42) and a 2D laparoscopic group (N = 113). The 3D laparoscopic group was further divided into two subgroups based on age-Group 1 (age ≤12 months, N = 11) and Group 2 (age >12 months, N = 31). We analyzed data in terms of the following characteristics: patient demographics, perioperative and follow-up conditions, and complications. Moreover, we also recorded and analyzed the surgeon's assessment on laparoscopic system usage. Results: Cyst excision and Roux-en-Y hepaticojejunostomy (HJ) was successfully completed in all the patients. Intergroup differences in operating time and blood loss were statistically significant. There were no significant differences between the two groups in early and late complications, such as the incidence of wound infection, HJ stricture, or adhesive ileus. There were no significant differences in the operative data and outcomes between the two subgroups of patients who underwent 3D laparoscopic treatment. Based on surgeon's assessment, the 3D laparoscopic system had better depth perception and accuracy than the 2D laparoscopic system; however, there was no difference in the adverse effect on surgeons. Conclusions: Compared with the traditional 2D laparoscopic surgery, 3D laparoscopic surgical resection of CCCs combined with jejunum Roux-en-Y anastomosis is a safer and more effective procedure that can shorten operative time and reduce intraoperative bleeding with no increase in surgical strain. The 3D laparoscopic surgery technique may provide a better choice for CCC operations.

Long-term outcomes of laparoscope-assisted heart-shaped anastomosis for children with hirschsprung disease: A 10-year review study.

BACKGROUND AND OBJECTIVE: Several operating procedures have been reported for treating Hirschsprung Disease (HD), but the incidence of postoperative complications remains at a high level affecting both the patients' prognosis and postoperative life quality. Heart-shaped anastomosis as a novel surgery method for children with HD has been improved by our medical center. This new surgery method is characterized by a heart-shaped colorectal anastomosis after splitting the posterior rectum wall to 0.5 cm above the dentate line. The aim of this review was to research the outcomes and quality of life of laparoscope-assisted heart-shaped anastomosis (LHSA) for children with HD by comparing it with a more generally applied surgery method, the laparoscope-assisted Soave procedure (LSP). METHODS: A retrospective review was conducted for 198 patients who underwent operations from January 2005 to December 2014 in our institution, who were divided into 97 cases of the LHSA group and 101 cases of LSP group according to the treatment methods; all the outcomes and individual quality of life data were assessed and compared. RESULT: All the enrolled 198 children diagnosed with HD, LHSA and LSP had been successfully completed in all cases. For complications, the incidence of constipation and soiling in the LHSA group was lower than the LSP group (P = 0.030, P = 0.042 respectively). On aspects of quality of life after operation, the individual quality of life of children with fecal incontinence was scored; and the higher the scores, the better the quality of life. Patients in the LHSA group had higher scores in terms of soiling, unhappy or anxious and peer rejection than the LSP group (P = 0.003, P = 0.009, P = 0.021, respectively). Other surgical characteristics and outcomes did not significantly differ between the two groups. CONCLUSION: LHSA is a feasible and safe minimally invasive surgery method with good long-term follow-up outcomes for HD patients. Compared with LSP, LHSA has advantages of low incidence of constipation and soiling, and provides better quality of life. Therefore, LHSA may provide a better choice for HD patients. LEVELS OF EVIDENCE: The type of study was Clinical Research Paper and the level of evidence was level III.

Results of rectoanal manometry after a novel laparoscopic technique: laparoscope-assisted heart-shaped anastomosis for Hirschsprung's disease.

PURPOSE: The present research utilizes a mid-term follow-up study to assess the results of anorectal manometry after laparoscope-assisted heart-shaped anastomosis (LHSA) for Hirschsprung's disease (HSCR), and compares it to a more generally applied approach, the laparoscope-assisted Soave procedure (LSP). METHODS: Retrospectively, patients from January 2015 to June 2017 who received LHSA or LSP were included in this study. After surgery, anorectal manometry was performed by the outpatient department. Anal sphincter resting pressure, anal canal length, amplitude of anal contraction, and frequency of anal contraction pre- and postoperatively were recorded. Additionally, mid-term complications were also monitored. RESULTS: Preoperative manometry showed no statistically significant difference between the LHSA and LSP groups. Postoperatively, anal sphincter resting pressure was lower in the LHSA group (60.64 ± 9.33 vs. 68.84 ± 11.80 mmHg, p = 0.001). Furthermore, anal canal length of the LHSA group was shorter than that of the LSP group (1.41 ± 0.18 vs. 1.53 ± 0.25 cm, p = 0.015). Frequency of anal contraction also showed a statistically significant difference between the LHSA and LSP groups (13.53 ± 2.17 vs. 12.50 ± 2.03 per minute, p = 0.032). The complication rates showed no significant difference and were as follows: incidence of enterocolitis was 13.89% in the LHSA group and 20.45% in the LSP group, incidence of constipation was 11.11% after LHSA and 27.27% after LSP, and incidence of soiling was 13.89% after LHSA and 25.00% after LSP. CONCLUSIONS: Manometric results of this study show satisfactory outcomes after LHSA. LHSA is an advanced surgical technique to make intestinal anastomosis easy and ensure a good prognosis.

Intravenous immunoglobulin for the treatment of intractable cholangitis after Kasai portoenterostomy in biliary atresia patients.

OBJECTIVE: To assess the efficacy of intravenous immunoglobulin (IVIG) as add-on treatment for intractable cholangitis (IC) after Kasai portoenterostomy (KPE) in biliary atresia (BA) patients. METHODS: 113 BA patients who had one or more episodes of cholangitis after KPE were recruited in this study. According to whether response to routine conservative treatment, all patients were divided into IC group and simple cholangitis (SC) group. Meanwhile, patients with IC subdivided into IVIG group and control group according to whether application of IVIG. RESULTS: The IC group had higher serum procalcitonin (PCT) (P = 0.014), C-reactive protein (CRP) (P = 0.023), and γ-Gltamyltranspeptidase (γ-GGT) (P = 0.031) level than the SC group. The IVIG group had shorter duration of fever after treatment (P = 0.011) and length of hospital stay (P = 0.018) than the control group. The time until recurrent episode of cholangitis was significant longer in IVIG group than in control group (P = 0.019). CONCLUSIONS: IVIG as add-on treatment may be an effective treatment for the cholangitis acute episode, and we conclude by calling for more prospective studies to attest to the role of IVIG in the treatment of cholangitis.