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.

Whole exome sequencing applied to 42 Han Chinese patients with posterior hypospadias.

Hypospadias, a malformation of male external genitalia, is characterized by an aberrant opening of the urethra on the ventral side of the penis. It is considered a complex disorder with both environmental and genetic factors involved in its pathogenesis. To identify the genetic abnormality involved in the pathogenesis of hypospadias, we performed whole exome sequencing (WES) analysis in 42 hypospadias patients with karyotype 46, XY in the Nanhai Meternity&Child Health Hospital of Foshan. All the likely pathogenic variants were confirmed by Sanger sequencing and assessed by Sorting Intolerant from Tolerant (SIFT), PROVEAN, PolyPhen2, ClinPred, LRT, Mutation Assessor, FATHMM, and GERP software. We discovered 27 gene mutations in 20 patients, including eight cases of the SRD5A2 gene, 4 cases of the AR gene, 3 cases of the CYP17A1 gene, 1 case of the WT1 gene, 1 case of the ANOS1 gene, 1 case of the NR5A1 gene, 1 case of the FGFR1 gene, and one case of the DHX37 gene. Our study is the first to describe six novel missense mutations, AR(c.302G > A, c.2593G > T, and c.1705G > T), CYP17A1(c.1298 T > C), FGFR1 (c.995C > T) and DHX37(c.923G > A). In summary, genetic defect detection was useful for early diagnosis of severe hypospadias in the Han Chinese population. Nevertheless, most cases remain unexplained, and the exact pathogenesis of hypospadias still needs further study.

Clinical characteristics of necrotizing pneumonia caused by Mycoplasma pneumoniae infection in children / 临床儿科杂志

Objectives To analyze the clinical characteristics, treatment and prognosis of necrotizing pneumonia caused by Mycoplasma pneumoniae (MP) infection in children. Method The clinical data of children with necrotizing pneumonia cause by MP infection from October 2016 to October 2017 were retrospectively analyzed. Results A total of 26 children (10 males and 16 females) with an average age of (5.76±2.60) years, were enrolled in the study. All children were characterized by fever and cough. High fever ( ≥ 39.0 ℃) was seen in 23 cases (88.5％) and the total duration of fever was (16.88±7.42) days. Pulmonary auscultation showed a reduction in respiratory sounds in all children. The range of peripheral blood leukocytes were (9.0～36.8) ×109/L, mean peak neutrophil ratio was (69.2±13.2) ％, and the range of C-reactive protein (CRP) was (1～202.5) mg/L. The mean value of lactic dehydrogenase (LDH) was (448±247) U/L. At the beginning of the disease, the chest images showed homogeneous solid high-density images over the whole lung lobe and 20 cases (76.9％) were complicated with pleural effusion. At the later stage, lung CT showed thin-walled cavities or multiple air-containing cysts on the basis of lung consolidation. Fiberoptic bronchoscopy showed lumen obstruction caused by mucus plugs in 23 cases (88.5％) . All the children were treated with methylprednisolone. The dose of 2 mg/ (kg·d) was effective in 21 cases and the fever was relieved in 5 cases after the dose was adjusted to 4 mg/ (kg·d) , and the average hormone application time was (13.08 ± 8.38) d. The median length of hospital stay was [16.5 (7～32) ] d. Two cases were lost to follow-up and 24 cases finished 6-month follow-up. Lung CT showed almost complete recovery of the lungs in 16 cases, residual pleural hypertrophy in 5 cases, and bronchiectasis in 1 case and bronchiolitis obliterans in 2 cases. Conclusion Necrotic pneumonia in children caused by MP infection is characterized by persistent high fever, decreased respiratory sounds, lung consolidation and mucus plugs induced lumen obstruction. The prognosis is relatively good after active anti-infection and hormone therapy.

Analysis on viral pathogens in hospitalized children with acute respiratory tract infection from 2014 to 2017 in Shijiazhuang / 中华实验和临床病毒学杂志

Objective@#To investigate the epidemiologic features of respiratory viral etiology in hospitalized children with acute respiratory tract infection (ARTI) in Shijiazhuang.@*Methods@#A total of 28 512 cases of hospitalized children with clinical diagnosis of ARTI in Children′ s Hospital of Hebei Province from 2014 to 2017 were recruited into this study. One nasopharyngeal swab was collected from each patient. Immunofluorescence assay was used to detect seven kinds of respiratory viruses, including respiratory syncytial virus (RSV), parainfluenza virus (PIV) type 1-3, influenza virus type A, B (FluA, FluB) and adenovirus (ADV).@*Results@#At least one viral pathogen was identified in each of 9 263 out of 28 512 patients and the overall positive rate was 32.5%. Of 9 263 virus-positive patients, 9 070 (97.9%) had mono-infection. The most frequently detected virus was RSV, followed by PIV-3 and FluA. The positive rates of RSV and PIV-1 showed annually decreasing tendency, meanwhile the positive rate of FluA increased in the nearly two years. The detection rate of ADV and PIV-1 increased every other year. There was a significant difference in the positive rate among different years (P<0.05). The overall positive rate decreased along with the age increased (linear by linear association χ2=1191.289, P<0.05). The detection rates of RSV and PIV-3 were the highest in groups of <1 year old and 1-3 years old and decreased along with the age increased. The preschool children were more susceptible to developing FluA, FluB and ADV related diseases. There was a significant difference in the positive rate among different age groups (P<0.05). The viral distribution was uneven in different seasons, and the infection peaked in winter, the difference was statistically significant (P<0.05). The epidemic seasons of RSV and FluA were winter, and FluB infection was epidemic in winter and spring. The positive rates of PIV-1 and PIV-2 were most common in summer and autumn. PIV-3 was usually prevalent in spring and summer and ADV was prevalent sporadically.@*Conclusions@#RSV is the most common pathogen in hospitalized children with clinical diagnosis of ARTI during 2014-2017 and the positive rate of which showed an annually decreasing tendency. The positive rate of FluA increased in the nearly two years. Children in infancy are susceptible to the seven common respiratory viruses and winter is the epidemic season for these viruses.

4.4 Å Resolution Cryo-EM structure of human mTOR Complex 1

Mechanistic target of rapamycin (mTOR) complex 1 (mTORC1) integrates signals from growth factors, cellular energy levels, stress and amino acids to control cell growth and proliferation through regulating translation, autophagy and metabolism. Here we determined the cryo-electron microscopy structure of human mTORC1 at 4.4 Å resolution. The mTORC1 comprises a dimer of heterotrimer (mTOR-Raptor-mLST8) mediated by the mTOR protein. The complex adopts a hollow rhomboid shape with 2-fold symmetry. Notably, mTORC1 shows intrinsic conformational dynamics. Within the complex, the conserved N-terminal caspase-like domain of Raptor faces toward the catalytic cavity of the kinase domain of mTOR. Raptor shows no caspase activity and therefore may bind to TOS motif for substrate recognition. Structural analysis indicates that FKBP12-Rapamycin may generate steric hindrance for substrate entry to the catalytic cavity of mTORC1. The structure provides a basis to understand the assembly of mTORC1 and a framework to characterize the regulatory mechanism of mTORC1 pathway.

The PHD1 finger of KDM5B recognizes unmodified H3K4 during the demethylation of histone H3K4me2/3 by KDM5B

KDM5B is a histone H3K4me2/3 demethylase. The PHD1 domain of KDM5B is critical for demethylation, but the mechanism underlying the action of this domain is unclear. In this paper, we observed that PHD1KDM5B interacts with unmethylated H3K4me0. Our NMR structure of PHD1KDM5B in complex with H3K4me0 revealed that the binding mode is slightly different from that of other reported PHD fingers. The disruption of this interaction by double mutations on the residues in the interface (L325A/D328A) decreases the H3K4me2/3 demethylation activity of KDM5B in cells by approximately 50% and increases the transcriptional repression of tumor suppressor genes by approximately twofold. These findings imply that PHD1KDM5B may help maintain KDM5B at target genes to mediate the demethylation activities of KDM5B.

Effects of Hirudo extract on release of thromboxane B2 and 6-keto-prostaglandin F1αfrom cultured human umbilical vein endothelial cells induced by thrombin / 中国中西医结合急救杂志

10.3969/j.issn.1008-9691.2013.03.007

Kinetic changes of C-reactive protein and carotid intima-media thickness in patients with IGT-3 years prospective study / 中国医师杂志

Objective To investigate the relationship between C-reactive protein (CRP) and carotid intima-media thickness in pa-tients with impaired glucose tolerance (IGT). Methods Senan CRP was measured with immunoturhidimetry and the carotid intima-media thickness (CAIMT) was measured using color Doppler in 108 patients with impaired glucose tolerance (IGT) and 80 subjects with normal glucose tolerance(NGT).Then we observed all IGT patients for 3 years using prospective follow-up method, Oral Glucose tolerance test (OGGT) and every index were measured in follow-up 1.5 year and 3 year. Results 2 objects were lost to follow-up. IGT group showed a significant higher CAIMT and CRP compared with NGT group. After follow-up 1.5 year and 3 year, the patients with impaired glucose toler-ance (IGT) were divided into type 2 diabetes (T2DM) group and IGT group based on the level of blood glucose. Both T2DM group and IGT group showed a significant higher CAIMT and CRP, compared with NGT group. The level of serum CRP of T2DM group was higher than that of IGT group, and the level of serum CRP of IGT group was higher than that of NGT group. There were great differences between each group.Linear correlation showed that the level of blood glucose was positively correlated with CRP and CAIMT in T2DM group after follow-up 3 year. CAIMT was positively correlated with the level of blood glucose and CRP. Mulfivariant stepwise regression showed that CRP was signifi-canfly correlated with the level of blood glucose and CAIMT. Conclusion Inflammation played an important role in the development of dia-betes, and it had great vessels complication. The patients with impaired glucose tolerance, who have high level of CRP, were facilitated to be diabetes, and they were at risk of getting great vessels complication during the phase of impaired glucose tolerance. So it would be helpful to prevent IGT patients with high CRP or CAIMT with anti-inflammatory therapy.

Effects of deliberate hypotension combined with acute isovolemic hemodilution on glomerular filtration rate in rabbits / 中华麻醉学杂志

Objective To determine the effects of acute normovolemic hemodilution combined with deliberate hypotension induced by sodium nitroprusside on glomerular filtration rate. Methods Forty rabbits weighing 2.6-4.0kg were anesthetized with intramuscular ketamine(1mg?kg-1). The animals were tracheotomized and intubated.Anesthesia was maintained with 0.5% halothane inhalation and fentanyl infusion (2?g?kg-1?min-1 ) . Carotid artery was cannulated for blood pressure monitoring and blood gas analysis and external jugular vein was cannulated for CVP monitoring and blood sampling. The animals were randomly allocated to one of four groups of ten animals in each group: control group; deliberate hypotension group;hemodilution group and combined hemodilution-deliberate hypotension group. Deliberate hypotension was induced with 0.02% sodium nitroprusside and MAP was reduced to 35-40mm Hg. In hemodilution group and combined group blood was removed from artery until Hct was reduced to 20 % -25 % and normovolemia was maintained by simultaneous infusion of Ringer's lactate solution, 3 times the volume of blood removed. Glomerular filtration rate was assessed by inulin clearance rate. 1% inulin 3 ml was injected I. V. over 30 s and blood samples were taken from CVP line for determination of blood inulin concentration at 1,3,5, 10, 30, 60, and 100 min after injection of inulin. Results Urine output decreased significantly in hypotension group and increased in hemodilution group. There was no significantly difference in urine output between control and combined groups. Inulin clearance decreased significantly in hypotension group as compared with that in control group, but there was no significantly difference in inulin clearance between control and combined groups. Conclusions Deliberate hypotension can decrease renal blood flow and deliberate hypotension combined with hemodilution can maintain kidney microcirculation.