RESUMO
Background: The semaphorin 3D (SEMA3D) gene has been implicated in the pathogenesis of Hirschsprung disease (HSCR), a complex genetic disorder characterized by the loss of ganglion cells in varying lengths of gastrointestinal tract. We wished to investigate the role of SEMA3D variants, both rare and common variants, as well as its mRNA expression in Indonesian HSCR patients. Methods: Sanger sequencing was performed in 54 HSCR patients to find a pathogenic variant in SEMA3D. Next, we determined SEMA3D expression in 18 HSCR patients and 13 anorectal malformation colons as controls by quantitative real-time polymerase chain reaction (qPCR). Results: No rare variant was found in the SEMA3D gene, except one common variant in exon 17, p.Lys701Gln (rs7800072). The risk allele (C) frequency at rs7800072 among HSCR patients (23%) was similar to those reported for the 1,000 Genomes (27%) and ExAC (28%) East Asian ancestry controls (p = 0.49 and 0.41, respectively). A significant difference in SEMA3D expression was observed between groups (p = 0.04). Furthermore, qPCR revealed that SEMA3D expression was strongly up-regulated (5.5-fold) in the ganglionic colon of HSCR patients compared to control colon (ΔCT 10.8 ± 2.1 vs. 13.3 ± 3.9; p = 0.025). Conclusions: We report the first study of aberrant SEMA3D expressions in HSCR patients and suggest further understanding into the contribution of aberrant SEMA3D expression in the development of HSCR. In addition, this study is the first comprehensive analysis of SEMA3D variants in the Asian ancestry.
RESUMO
BACKGROUND: Hirschsprung-associated enterocolitis (HAEC) is the most common complication of Hirschsprung disease (HSCR) that may happen pre-operatively. Several methods have been reported to determine HAEC. Because the diagnosis of pre-operative HAEC might change the surgical plan, we aimed to determine the accuracy of the classical criteria for diagnosis of pre-operative HAEC and using the Delphi method as a gold standard. METHODS: Medical records of HSCR children who were admitted to our hospital from January 2009 to December 2015 were retrospectively analyzed. RESULTS: Ninety-six subjects were involved in this study, consisting of 74 males and 22 females. The most common findings of the Delphi score were abdominal distension (100%) and dilated loops of bowel (100%), followed by leucocytosis (78.6%), lethargy (71.4%), cutoff sign in rectosigmoid with absence of distal air (71.4%), and shift to left (71.4%). The frequency of pre-operative HAEC was 4.2% and 14.6% using the classical criteria and Delphi method, respectively (p = 0.016). The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy rates of the classical criteria for diagnosis of pre-operative HAEC were 14.3% (95% CI: 1.8-42.8%), 97.6% (95% CI: 91.5-99.7%), 50% (95% CI: 13.3-86.7%), 87% (95% CI: 84.3-89.2), and 85.4% (95% CI: 76.7-91.8%), respectively. CONCLUSIONS: The frequency of pre-operative HAEC is low in our hospital. The accuracy of the classical criteria is considered relatively moderate for diagnosis of pre-operative HAEC.
