Integrated machine learning-driven disulfidptosis profiling: CYFIP1 and EMILIN1 as therapeutic nodes in neuroblastoma.

BACKGROUND: Neuroblastoma (NB), a prevalent pediatric solid tumor, presents formidable challenges due to its high malignancy and intricate pathogenesis. The role of disulfidptosis, a novel form of programmed cell death, remains poorly understood in the context of NB. METHODS: Gaussian mixture model (GMM)-identified disulfidptosis-related molecular subtypes in NB, differential gene analysis, survival analysis, and gene set variation analysis were conducted subsequently. Weighted gene co-expression network analysis (WGCNA) selected modular genes most relevant to the disulfidptosis core pathways. Integration of machine learning approaches revealed the combination of the Least absolute shrinkage and selection operator (LASSO) and Random Survival Forest (RSF) provided optimal dimensionality reduction of the modular genes. The resulting model was validated, and a nomogram assessed disulfidptosis characteristics in NB. Core genes were filtered and subjected to tumor phenotype and disulfidptosis-related experiments. RESULTS: GMM clustering revealed three distinct subtypes with diverse prognoses, showing significant variations in glucose metabolism, cytoskeletal structure, and tumor-related pathways. WGCNA highlighted the red module of genes highly correlated with disulfide isomerase activity, cytoskeleton formation, and glucose metabolism. The LASSO and RSF combination yielded the most accurate and stable prognostic model, with a significantly worse prognosis for high-scoring patients. Cytological experiments targeting core genes (CYFIP1, EMILIN1) revealed decreased cell proliferation, migration, invasion abilities, and evident cytoskeletal deformation upon core gene knockdown. CONCLUSIONS: This study showcases the utility of disulfidptosis-related gene scores for predicting prognosis and molecular subtypes of NB. The identified core genes, CYFIP1 and EMILIN1, hold promise as potential therapeutic targets and diagnostic markers for NB.

miR-375-3p targets YWHAB to attenuate intestine injury in neonatal necrotizing enterocolitis.

PURPOSE: Necrotizing enterocolitis (NEC) is a significant contributor to neonatal mortality. This study aimed to investigate the role of high levels of miR-375-3p in breast milk in the development of NEC and elucidate its mechanism. METHODS: Differential expression of miR-375-3p in the intestines of breast-fed and formula-fed mice was confirmed using real-time polymerase chain reaction (RT-PCR). NEC mice models were established, and intestinal injury was assessed using HE staining. RT-PCR and Western blot were conducted to examine the expression of miR-375-3p, tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein ß (YWHAB), as well as the inflammatory in IEC-6 cells, and intestinal tissues obtained from NEC mice and patients. Flow cytometry and cell counting kit-8 (CCK-8) were employed to elucidate the impact of miR-375-3p and YWHAB on cell apoptosis and proliferation. RESULTS: Breastfeeding increases miR-375-3p expression in the intestines. The expression of miR-375-3p in NEC intestinal tissues exhibited a significant decrease compared to the healthy group. Additionally, the expression of interleukin-6 (IL-6), interleukin-10 (IL-10), and tumor necrosis factor-α (TNF-α) was higher in the NEC group compared to the control group. Down-regulation of miR-375-3p inhibited IEC-6 cell proliferation, increased apoptosis, and elevated secretion of inflammatory factors. Bioinformatics revealed that YWHAB may be a target of miR-375-3p. RT-PCR and Western blot indicated a down-regulation of YWHAB expression in intestines of NEC patients and mice. Furthermore, YWHAB was found to be positively connected with miR-375-3p. Knockdown miR-375-3p down-regulated YWHAB expression in cells. Inhibition of YWHAB exhibited similar effects to miR-375-3p in IEC-6 cells. YWHAB plasmid partially reverse cellular functional impairment induced by miR-375-3p knockdown. CONCLUSIONS: Breastfeeding elevated miR-375-3p expression in intestines in neonatal mice. MiR-375-3p leads to a decrease in apoptosis of intestinal epithelial cells, an increase in cell proliferation, and a concomitant reduction in the expression of inflammatory factors partly through targeting YWHAB.

Circular RNA MTCL1 targets SMAD3 by sponging miR-145-5p for regulation of cell proliferation and migration in Hirschsprung's disease.

BACKGROUND: Hirschsprung's disease (HSCR) is a congenital disorder resulting from abnormal development of the enteric nervous system (ENS). Given the complexity of its pathogenesis, it is important to investigate the role of epigenetic inheritance in its development. As Circ-MTCL1 is abundant in brain tissue and colon tissue, whether it has a significant part in the development of ENS is worth exploring. This study clarifies its role in HSCR and identifies the specific molecular mechanisms involved. METHODS: Diseased and dilated segment colon tissues diagnosed as HSCR were collected for the assessment of gene expression levels using RT-PCR. EdU and CCK-8 assays were adopted to evaluate cell proliferation, and Transwell assay was adopted to assess cell migration. The interaction between Circ-MTCL1, miR-145-5p and SMAD3 was confirmed by dual luciferase reporter gene analysis, RT-PCR and Western blotting. RESULTS: Circ-MTCL1 was down-regulated in the aganglionic colon tissues. The decreased expression of Circ-MTCL1 associated with a reduction in cell migration and proliferation. Bioinformatics analysis and cellular experiments confirmed its role might have been associated with the inhibition of miR-145-5p. MiR-145-5p was up-regulated in HSCR diseased segment colon tissues, exhibiting a negative correlation with Circ-MTCL1. Overexpression of miR-145-5p reversed the inhibition of cell migration and proliferation associated with Circ-MTCL1 down-regulation. The expression of SMAD3 was inhibited by miR-145-5p. The overexpression of SMAD3 eliminated the miR-145-5p-associated inhibition of cell migration and proliferation. Overexpression of miR-145-5p reversed the inhibitory effects of Circ-MTCL1 down-regulation-associated inhibition of cell migration and proliferation, while suppressing SMAD3 expression. Conversely, overexpression of SMAD3 counteracted the miR-145-5p-associated inhibition of cell migration and proliferation. CONCLUSIONS: Circ-MTCL1 may function as a miR-145-5p sponge, regulating the expression of SMAD3 and influencing cell migration and proliferation, thus participating in the development of HSCR.

Analysis of variant of GLI3 gene in a child featuring autosomal dominant Pallister-Hall syndrome / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical and genetic characteristics of a child with Pallister-Hall syndrome (PHS).@*METHODS@#DNA was extracted from peripheral blood sample from the child and subjected to whole exome sequencing. Suspected variants were verified by Sanger sequencing of his family members.@*RESULTS@#Genetic testing revealed that the child has harbored a heterozygous c.3320_3330delGGTACGAGCAG (p.G1107Afs×18) variant of the GLI3 gene. Neither parent was found to carry the same variant.@*CONCLUSION@#The c.3320_3330delGGTACGAGCAG (p.G1107Afs×18) frameshift variant of the GLI3 gene probably underlay the pathogenesis of PHS in this child. Genetic testing should be considered for patients featuring hypothalamic hamartoma and central polydactyly.

Comparison of efficacy of different concentrations of ropivacaine for interscalene brachial plexus block in patients undergoing arthroscopic shoulder surgery under general anesthesia / 中华麻醉学杂志

Objective:To compare the efficacy of different concentrations of ropivacaine for interscalene brachial plexus block in patients undergoing arthroscopic shoulder surgery under general anesthesia.Methods:Ninety American Society of Anesthesiologists physical statusⅠor Ⅱ patients (NYHA classⅠorⅡ) of both sexes, aged 18-64 yr, with body mass index of 18.0-26.9 kg/m 2, undergoing elective arthroscopic shoulder surgery were selected, and were divided into 3 groups ( n=30 each) using a random number table method: 0.25% ropivacaine group (group A), 0.375% ropivacaine group (group B) and 0.5% ropivacaine group (group C). Interscalene brachial plexus block was performed with 0.25%, 0.375% and 0.5% ropivacaine 20 ml in A, B and C groups, respectively.Before operation (T 0) and at 30 min (T 1), 4 h (T 2), 6 h (T 3), 8 h (T 4), 10 h (T 5) and 12 h (T 6) after administration, the diaphragmatic mobility was measured and recorded using M-mode ultrasound and forced expiratory volume in the first second (FEV 1) and forced vital capacity (FVC) were measured using portable spirometer.The occurrence of phrenic paralysis was recorded at T 1-6.The duration of sensory and motor block was recorded.When visual analogue scale score>3 within 24 h after operation, flurbiprofen axetil 50 mg was injected intravenously for analgesia and the consumption was recorded.The adverse reactions such as cardiovascular events, local anesthetic intoxication, Horner syndrome, pneumothorax, and nausea and vomiting within 24 h after administration were recorded. Results:Compared with group A, the diaphragmatic mobility was significantly decreased during quiet breathing at T 1-3 and was decreased during deep breathing at T 2-5, and the diaphragmatic paralysis rate was increased during quiet and deep breathing at T 2-3 in group B, diaphragmatic mobility was decreased during quiet and deep breathing at T 1-6, diaphragmatic paralysis rate was increased during quiet and deep breathing at T 1-4, FEV 1% and FVC% were decreased at T 1 and FVC% was decreased at T 2 in group C, and the duration of sensory and motor block was prolonged in B and C groups ( P<0.05 or 0.01). Compared with group B, the diaphragmatic mobility was significantly decreased during quiet breathing at T 4-6 and was decreased during deep breathing at T 1-6, the diaphragmatic paralysis rate during quiet breathing was increased at T 2-4 ( P<0.05) was increased during deep breathing at T 3-4, and FEV 1 % and FVC % at T 1 were decreased in group C ( P<0.05). There was no significant difference in the postoperative requirement for flurbiprofen axetil and the incidence of adverse reactions within 24 h after administration among the 3 groups ( P>0.05). Conclusion:0.25% ropivacaine 20ml provides better efficacy when used for interscalene brachial plexus block in the patients undergoing arthroscopic shoulder surgery.