[Analysis of a child with Verheij syndrome due to variant of PUF60 gene].

OBJECTIVE: To explore the clinical phenotype and genetic variant in a child with Verheij syndrome (VRJS). METHODS: A child who had presented at the Soochow University Affiliated Children's Hospital and Wujiang District Children's Hospital in July 2022 for "elevated scapula since early childhood" was selected as the study subject. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing. Candidate variant was verified by Sanger sequencing and bioinformatic analysis. RESULTS: The child had manifested elevated scapulae, torticollis, neck asymmetry, facial dysmorphism, dispersed café-au-lait spots, limited mobility of upper limbs and shoulder joints, and intellectual disability. Sequencing revealed that he has harbored a de novo heterozygous c.405dupT (p.Ile136Tyrfs*4) variant of the PUF60 gene. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), this variant was classified as pathogenic (PVS1+PS2_moderate+PM2_supporting). Combined his clinical features and result of genetic testing, the child was diagnosed with VRJS due to variant of the PUF60 gene. CONCLUSION: The clinical manifestations of VRJS include facial dysmorphism, intellectual disability, elevated scapulae, vertebral fusion, other skeletal malformations, without significant abnormalities of the heart, kidney, and eyes, which need to be distinguished from Klippel-Feil syndrome. Above finding has expended the mutation spectrum of the PUF60 gene and provided a reference for delineation of the genotype-phenotype correlation of the VRJS.

Development of a nomogram for severe influenza in previously healthy children: a retrospective cohort study.

OBJECTIVE: We aimed to develop a nomogram to predict the risk of severe influenza in previously healthy children. METHODS: In this retrospective cohort study, we reviewed the clinical data of 1135 previously healthy children infected with influenza who were hospitalized in the Children's Hospital of Soochow University between 1 January 2017 and 30 June 2021. Children were randomly assigned in a 7:3 ratio to a training or validation cohort. In the training cohort, univariate and multivariate logistic regression analyses were used to identify risk factors, and a nomogram was established. The validation cohort was used to evaluate the predictive ability of the model. RESULT: Wheezing rales, neutrophils, procalcitonin > 0.25 ng/mL, Mycoplasma pneumoniae infection, fever, and albumin were selected as predictors. The areas under the curve were 0.725 (95% CI: 0.686-0.765) and 0.721 (95% CI: 0.659-0.784) for the training and validation cohorts, respectively. The calibration curve showed that the nomogram was well calibrated. CONCLUSION: The nomogram may predict the risk of severe influenza in previously healthy children.

Disulfide-stapled design of α-helical bundles to target the trimer-of-hairpins motif of human respiratory syncytial virus fusion protein.

Human respiratory syncytial virus (RSV) is the major cause of acute lower respiratory tract infections worldwide in infants and young children. The RSV F glycoprotein is a class I fusion protein that mediates viral entry into host cells and is a major target of neutralizing antibodies. Targeting F glycoprotein has been recognized as a promising antiviral therapeutic strategy against RSV infection. Here, we reported the disulfide-stapled design of α-helical bundle to target the trimer-of-hairpins (TOH) motif of RSV F glycoprotein, which is the central regulatory module that triggers viral membrane fusion event. In TOH motif, three N-terminal heptad repeat (NtHR) helices form a trimeric coiled-coil core and other three C-terminal heptad repeat (CtHR) helices add to the core in an antiparallel manner. Interaction analysis between NtHR and CtHR revealed that the C-terminal tail of CtHR packs tightly against NtHR as compared to the N-terminal and middle regions of CtHR. A core binding site in CtHR C-terminus was identified, which represents a 13-mer chp peptide and can effectively interact with NtHR helix in native ordered conformation but would become largely disordered when splitting from the protein context of CtHR helix. Two chp helices were stapled together in a parallel manner with single, double or triple disulfide bridges, thus systematically resulting in seven disulfide-stapled α-helical bundles. Molecular simulations revealed that the double and triple stapling can effectively stabilize the structured conformation of α-helical bundles, whereas the free conformation of single-stapled bundles still remain intrinsically disordered in solvent. The double-stapled bundle chp-ds[508,516] and the triple-stapled bundle chp-ts[508,512,516] were rationally designed to have high potency; they can form a tight three-helix bundle with NtHR helix, thus potently targeting NtHR-CtHR interactions involved in RSV-F TOH motif through a competitive disruption mechanism.

Risk factors for delayed radiographic resolution in children with refractory Mycoplasma pneumoniae pneumonia.

OBJECTIVE: To determine the risk factors for delayed radiographic resolution in children with refractory Mycoplasma pneumoniae pneumonia (RMPP) and explore the most suitable time for interventional bronchoscopy. METHODS: This retrospective study involved 142 children with RMPP who were admitted to our hospital from 1 January 2015 to 31 December 2017. They were divided into a common resolution group and a delayed resolution group based on their chest radiograph series. RESULTS: Among the 142 patients, 67 showed common resolution on chest radiographs and 75 showed delayed resolution. Independent risk factors for delayed resolution were a clinical course of ≥11.5 days before the performance of interventional bronchoscopy, mucus plug formation, corticosteroid resistance, and atelectasis. When bronchoscopy was performed before the disease had been present for <11.5 days, the length of hospitalization, total fever duration, and duration of time until disappearance of coughing were shorter than those in children who underwent bronchoscopy after the disease had been present for ≥11.5 days. CONCLUSIONS: Corticosteroid resistance, the time to interventional bronchoscopy, atelectasis, and mucus plug formation were associated with delayed resolution on chest radiographs. Performance of interventional bronchoscopy before the clinical course has reached 11.5 days may help alleviate clinical symptoms and improve radiographic resolution.