Anti-miR-141-3p maintains homeostasis between autophagy and apoptosis by targeting Yy1 in the fetal lumbosacral defecation center of rats.

Anorectal malformations (ARMs) are congenital diseases that lead to postoperative fecal incontinence, constipation, and soiling, despite improvements in surgery; however, their pathological mechanisms remain unclear. Here, we report the role of microRNA-141-3p in maintaining homeostasis between apoptosis and autophagy in the lumbosacral defecation center of fetal rats with ARMs. Elevated microRNA-141-3p expression inhibited YIN-YANG-1 expression by binding its 3' UTR, and repressed autophagy and triggered apoptosis simultaneously. Then, adenylate cyclase 3 was screened to be the downstream target gene of YIN-YANG-1 by chromatin immunoprecipitation sequencing experiments, and Yin Yang 1 could positively activate the transcription of adenylate cyclase 3 by directly interacting with the motif GAGATGG and ATGG in its promoter. Intraamniotic microinjection of adeno-rno-microRNA-141-3p-sponge-GFP in fetal rats with ARMs on embryonic day 15 restored apoptosis-autophagy homeostasis. These findings reveal that microRNA-141-3p upregulation impaired homeostasis between apoptosis and autophagy by inhibiting the YIN-YANG-1/adenylate cyclase 3 axis, and that intraamniotic injection of anti-microRNA-141-3p helped maintain homeostasis in the lumbosacral defecation center of ARMs during embryogenesis.

Therapeutic potential of adenovirus-encoding brain-derived neurotrophic factor for spina bifida aperta by intra-amniotic delivery in a rat model.

Spina bifida aperta is a type of neural tube defect (NTD). Although prenatal fetal surgery has been an available and effective treatment for it, the neurological functional recovery is still need to be enhanced. Our previous results revealed that deficiencies of sensory, motor, and parasympathetic neurons were primary anomalies that occurred with the spinal malformation. Therefore, we emphasized that nerve regeneration is critical for NTD therapy. We delivered an adenoviral construct containing genes inserted for green fluorescent protein and brain-derived neurotrophic factor (Ad-GFP-BDNF) into the amniotic fluid to investigate its prenatal therapeutic potential for rat fetuses with spina bifida aperta. Using immunofluorescence, TdT-mediated dUTP nick-end labeling staining, and real-time polymerase chain reaction analysis, we assessed cell apoptosis in the defective spinal cord and Brn3a positive neuron survival in the dorsal root ganglion (DRG); a protein array was used to investigate the microenvironmental changes of the amniotic fluid. We found that most of the overexpressed BDNF was present on the lesions of the spina bifida fetuses, the number of apoptosis cells in Ad-GFP-BDNF-transfected spinal cords were reduced, mRNA levels of Bcl2/Bax were upregulated and Casp3 were downregulated compared with the controls, the proportion of Brn3a positive neurons in DRG were increased by activating the BDNF/TrkB/Akt signaling pathway, and most of the significant changes in cytokines in the amniotic fluid were related to the biological processes of regulation of apoptotic process and generation of neurons. These results suggest that intra-amniotic Ad-GFP-BDNF gene delivery might have potential as a supplementary approach to treat congenital malformations of neural tubes.

Long-term outcome, colonic motility, and sphincter performance after Swenson's procedure for Hirschsprung's disease: a single-center 2-decade experience with 346 cases.

BACKGROUND: The aim of this study is to explore the long-term outcome and the colon and sphincter function in children undergoing the Swenson's operation for Hirschsprung's disease (HD). METHODS: Three hundred forty-six children (266 males and 80 females) undergoing Swenson's operation for HD for 8 to 20 years were followed up. Barium enema and defecography, total and segmental colonic transit time with the simplified radioopaque markers, and the anorectal vector manometry were used. RESULTS: Stooling patterns were fair in most patients, and the functional examinations were abnormal in few cases. CONCLUSIONS: The long-term outcome is satisfactory in most of the children undergoing the Swenson's operation for HD, but it is still not as good as what the surgeons have expected. This is probably because of the abnormal colonic motility and/or the dysfunctions of the internal anal sphincter.

Evaluation of anorectal function after transanal one-stage endorectal pull through operation in children with Hirschsprung's disease.

OBJECTIVE: The short-term efficacy of the transanal one-stage endorectal pull through operation for Hirschsprung's disease is satisfactory. However the long-term outcome of anorectal function has not been fully understood. The aim of this study was to evaluate the stooling pattern, colonic motility and anal sphincter performance after transanal one-stage pull through operation in children with Hirschsprung's disease. METHODS: Fifty-eight children who underwent transanal one-stage pull through operation for Hirschsprung's disease were followed up. The mean follow-up duration was 15.8 months (range, 6-24 months). The stooling patterns of the patients were investigated by the informed questionnaire. Barium enema, defecography, total and segmental colonic transit time and the anorectal vector manometry were performed. Thirty- three healthy children were used as controls. RESULTS: Most of patients had normal stool consistency and frequency. Postoperative enterocolitis occurred in 3 patients, and constipation was found in five patients. Postoperative soiling was observed in 9 patients. None of the 58 patients had incontinence, cuff infection, anastomotic leak and mortality. The barium enema showed that the configuration of the colon recovered well in most of patients. Postopertive defecography showed the anorectal angle of all the patients was open, fixed and bigger than that of preoperation and the healthy controls (P < 0.01). Postoperatively, the mean total gastrointestinal transit time (TGITT), the left colonic transit time (LCTT) and rectosigmoid colonic transit time (RSTT) in the 58 patients were significantly shorter than preoperatively (P < 0.01) and were similar to those of the control group. The rectoanal inhibitory reflex was regained in 5 patients. The anal maximal pressure of the patients with constipation in resting and squeezing condition were significantly higher than those of the asymptomatic patients and controls (P < 0.05). The vector volume (VV) and vector symmetric index (VSI) in patients with soiling were significantly lower than those in preoperation and the controls (P < 0.05). The VSI in the patients with constipation was significantly higher compared with the controls (P < 0.05). CONCLUSIONS: The stooling function, colonic motility and anal sphincter performance manifest well in most of the patients after the transanal endorectal pull through operation for Hirschsprung's disease. Stooling disorders in few cases are probably related to decrease or disappearance of the sigmoid loops, dysfunction of the "neorectosigmoid", an open and fixed anorectal angle and achalasia of the internal anal sphincter.

[Long-term outcomes of individualized biofeedback training based on the underlying dysfunction for patients with imperforate anus].

OBJECTIVE: Although better anorectal function was achieved due to the advanced operation procedures, the fecal incontinence and constipation still remain as major postoperative complications. The aim of this study was to design a individualized biofeedback program based on the underlying dysfunctions and to assess the short-term and long-term clinical and quality of life outcomes for patients with imperforate anus. METHODS: Thirty-one patients were treated using individualized biofeedback program according to their different dysfunctions after completed assessment of anorectal function. The many of special biofeedback training methods that consisted of the strengthening perianal muscles biofeedback, sensory discrimination biofeedback, synchronizing external sphincter contraction biofeedback and improving defecation dynamic biofeedback were selected to make up the individualized biofeedback training program. The clinical scores and quality of life scores were assessed at short-term and long-term follow-up. RESULTS: The clinical scores and quality of life scores in patients at before training group, after biofeedback training group, short-term follow-up group and long-term follow-up group were 3.2 +/- 1.1 and 8.7 +/- 1.1, 5.2 +/- 0.6 and 11.0 +/- 0.8, 5.0 +/- 0.6 and 10.8 +/- 0.9, and 4.6 +/- 0.7 and 10.0 +/- 0.7 respectively. After biofeedback training, the first leak volume and all parameters that evaluated the function of the perianal muscles were increased significantly, the abnormal rectal threshold sensations and defecation dynamics were reverted to normal, and 82% patients who have not external anal sphincter reflex acquired the new reflex. The latencies of pudendo-anal reflex have significant difference in patients with poor response compared with those patients with good response before and after the biofeedback training. At short-term follow-up 12 patients (57%) maintained the clinical outcome and only 9 patients (43%) have regressions slightly. At long-term follow-up 7 patients (33%) maintained the clinical outcome very well and 14 patients (67%) have regressions. Although the clinical and quality of life scores have decreased slightly at follow-up, they were still increased significantly compared with before biofeedback training group. CONCLUSION: The causes of postoperative fecal dysfunction in patients with imperforate anus are multifactorial, and the individualized biofeedback is more suitable for improvement of the clinical outcome and the quality of life, and it maintain good clinical outcome and quality of life at short-term and long-term follow-up.