Objective: To explore the relative importance of different food attributes and levels in food decision making of school aged children, and to understand their meal preferences, so as to provide the evidence for formulating precise intervention strategies for dietary behaviours of school aged children. Methods: From May to June 2024, a total of 854 children aged 11 to 15 years old were selected from 2 middle schools (each school in urban and rural areas) in both Hubei Province and Anhui Province by stratified cluster random sampling method to conduct a D-optimal discrete choice experiment. The mixed Logit model was used to analyze children s preference for meal attributes and different levels, and to calculate the relative importance (RI) of attributes and willingness to pay (WTP) in meal choices. Results: The included five food attributes had statistical significance on meal choice of school aged children ( P <0.05). The relative importance of food attributes affecting the meal choices of school aged children in descending order were dining mode ( RI =31.26%), food varieties ( RI =30.56%), cooking method( RI =23.84%), taste( RI =8.06%) and price ( RI =6.27%). Among them, school aged children preferred home cooked meals ( β =0.74) (WTP=86.3 yuan),varied foods(grain/tubers+vegetables+fish, meat, eggs and beans) ( β =0.61) (WTP=71.9 yuan), fried/roasted cooking ( β =0.51) and spicy taste ( β =0.33).Price was negatively correlated with meal choices( β =-0.01) ( P <0.05). Based on residential area and body mass index (BMI), the stratified analysis showed that dining mode was highest in the relative importance for rural children with overweight and obese children ( RI =31.28%,34.17%), both of whom preferred home cooked meals ( β =0.76, 0.91), and meals containing fish, meat, eggs and beans with grain/tubers or grain/tubers and vegetables in terms of food choice (area: β =0.53, 0.53 ; BMI: β =0.55, 0.56) ( P <0.05). Conclusions School aged children have different preferences for different attributes of meals. The quality of school meals should be improved,the cost of buying healthy meals should be reduced,targeted family health education should be carried out,and healthy cooking methods should be advocated.

The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines. Therefore, there remains a need for alternative antivirals that target processes less susceptible to mutations, such as the formation of six-helix bundle (6-HB) during the viral fusion step of host cell entry. In this study, a novel high-throughput screening (HTS) assay employing a yeast-two-hybrid (Y2H) system was established to identify inhibitors of HR1/HR2 interaction. The compound IMB-9C, which achieved single-digit micromolar inhibition of SARS-CoV-2 and its Omicron variants with low cytotoxicity, was selected. IMB-9C effectively blocks the HR1/HR2 interaction in vitro and inhibits SARS-CoV-2-S-mediated cell-cell fusion. It binds to both HR1 and HR2 through non-covalent interaction and influences the secondary structure of HR1/HR2 complex. In addition, virtual docking and site-mutagenesis results suggest that amino acid residues A930, I931, K933, T941, and L945 are critical for IMB-9C binding to HR1. Collectively, in this study, we have developed a novel screening method for HR1/HR2 interaction inhibitors and identified IMB-9C as a potential antiviral small molecule against COVID-19 and its variants.

Liver transplantation (LT) has become a standard treatment for end-stage liver diseases, and graft injury is intricately associated with poor prognosis. Granzyme B (GZMB) plays a vital role in natural killer (NK) cell biology, but whether NK-derived GZMB affects graft injury remains elusive. Through the analysis of single-cell RNA-sequencing data obtained from human LT grafts and the isolation of lymphocytes from mouse livers following ischemia-reperfusion injury (IRI), we demonstrated that 2NK cells with high expression of GZMB are enriched in patients and mice. Both systemically and liver-targeted depletion of NK cells led to a notable reduction in GZMB⁺ cell infiltration, subsequently resulting in diminished graft injury. Notably, the reconstitution of Il2rg ^-/- Rag2 ^-/- mice with purified Gzmb-KO NK cells demonstrated superior outcomes compared to those with wild-type NK cells. Crucially, global knockout of GZMB and pharmacological inhibition exhibited remarkable improvements in liver function in both mouse IRI and rat LT models. Moreover, a phosphorylated derivative of FDA-approved vidarabine was identified as an effective inhibitor of mouse GZMB activity by molecular dynamics, which could provide a potential avenue for therapeutic intervention. Therefore, targeting NK cell-derived GZMB during the LT process suggests potential therapeutic strategies to improve post-transplant outcomes.

Atopic dermatitis (AD) is a prevalent inflammatory skin disorder in which patients experience recurrent eczematous lesions and intense itching. The colonization of Staphylococcus aureus (S. aureus) is correlated with the severity of the disease, but its role in AD development remains elusive. Using single-cell RNA sequencing, we uncovered that keratinocytes activate a distinct immune response characterized by induction of Il24 when exposed to methicillin-resistant S. aureus (MRSA). Further experiments using animal models showed that the administration of recombinant IL-24 protein worsened AD-like pathology. Genetic ablation of Il24 or the receptor Il20rb in keratinocytes alleviated allergic inflammation and atopic march. Mechanistically, IL-24 acted through its heterodimeric receptors on keratinocytes and augmented the production of IL-33, which in turn aggravated type 2 immunity and AD-like skin conditions. Overall, these findings establish IL-24 as a critical factor for onset and progression of AD and a compelling therapeutic target.
Dermatitis, Atopic/genetics* ; Interleukins/metabolism* ; Animals ; Methicillin-Resistant Staphylococcus aureus/immunology* ; Mice ; Keratinocytes/microbiology* ; Humans ; Interleukin-33/immunology* ; Inflammation/microbiology* ; Staphylococcal Infections/microbiology* ; Disease Models, Animal ; Hypersensitivity/microbiology* ; Mice, Inbred C57BL

Objective To study the effects of monomers in Lagotis brachystachya Maxim including quercetin,echinacoside,and apigenin on the p38MAPK/JNK,TLR4/MyD88/NF-κB,and NLRP3 signaling through establishing a rat model of chronic alcoholic liver injury combined with gouty arthritis,and to explore the mechanism of the above-mentioned monomers in the treatment of chronic alcoholic liver injury combined with gouty arthritis.Methods Eighty rats were randomly divided into normal group,model group,bifendate group(100 mg·kg-1),colchicine group(0.3 mg·kg-1),and different dose groups of quercetin(50,25 mg·kg-1),echinacoside(50,25 mg·kg-1),and apigenin(50,25 mg·kg-1).The rats were administered at a dose of 10 mL·kg-1 per day in the morning and administered with 56-degree Red Star Erguotou at a dose of 4 mL·kg-1 in the afternoon,with gradual increasement of 2 mL·kg-1 per week until a final dose of 10 mL·kg-1.The rats were continuously fed by gavage for 8 weeks.Gouty arthritis model was induced at the 53th day of alcohol-based feeding.Toe volume was measured at difference time.At the end of the 8th week,blood was collected after the last administration.Serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP)levels,total cholesterol(TC)and triglycerides(TG)were measured.Enzyme-Linked Immunosorbent Assay(ELISA)was used to detect interleukin-1β(IL-1β)in rat serum.Western Blot was used to detect the expression of p38 mitogen-activated protein kinase(p38MAPK),phosphorylated p38MAPK(p-p38MAPK),c-Jun N-terminal kinase(JNK),phosphorylated JNK(p-JNK),Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),nuclear factor-κB(NF-κB),phosphorylated NF-κB(p-NF-κB),and NOD-like receptor protein domain containing 3(NLRP3)in liver and synovial tissues.Results Compared with the normal group,the toe swelling degree in the model group was significantly increased at difference time(P＜0.05,P＜0.01).The levels of ALT,AST,ALP,TC,TG,and IL-1β in serum were significantly increased(P＜0.01).The protein expression levels of p-p38MAPK,p-JNK,MyD88,p-NF-κB,TLR4,and NLRP3 in liver and synovial tissues were all increased to varying degrees(P＜0.01).Compared with the model group,the toe swelling degree in all treatment groups significantly decreased after 24 hours(P＜0.01).The levels of ALT and TC in the bifendate group and different dose groups of monomers decreased to varying degrees(P＜0.05,P＜0.01).Except for the low dose group of echinacoside,the levels of AST and ALP in other treatment groups decreased to varying degrees(P＜0.01).Except for the high dose group of quercetin,the levels of TG in other treatment groups decreased to varying degrees(P＜0.01).The protein expression levels of p-p38MAPK,p-JNK,MyD88,p-NF-κB,TLR4,and NLRP3 in liver and synovial tissues of rats in all treatment groups were down-regulated(P＜0.05,P＜0.01).Conclusion The monomers in Lagotis brachystachya Maxim show certain therapeutic effects of treating different diseases with same treatment on the rat model of chronic alcoholic liver injury combined with gouty arthritis,and their therapeutic effects may be mediated through p38MAPK/JNK,TLR4/MyD88/NF-κB,and NLRP3 signaling pathways.

Porcine parvovirus(PPV)is a causative agent of porcine parvovirus infection(PPI),which significantly impacts the pig industry due to its association with reproductive dysfunction.The condition is characterized by stillbirth,mummified fetuses,fetal weakness,and abortion in pregnant sows.The pathogenic mechanism remains incompletely understood,with no effective therapeutic drugs available currently.Despite extensive research on the host's response to PPV in-fection and identification of various signaling pathway transduction mechanisms,a comprehensive understanding is still lacking.This review aims to summarize the current knowledge regarding al-terations in related signaling pathways following PPV infection,provide novel insights into the pathogenesis of PPV and facilitate the drug development.

This study aims to investigate the effects of Japanese encephalitis virus(JEV)on TLRs signaling pathway and its regulation of the secretion of inflammatory factors during the infection of testicular interstitial cells,In this study,the mRNA levels of TLR3,TLR7,TLR8,TRIF and MyD88 genes were detected by qPCR after 1 MOI dose of JEV was inoculated into testicular stro-mal cells at different time periods.Western blot assay was used to detect the expression levels of TLR3,TLR7,TRIF and MyD88 protein at 6 h after JEV infection,and ELISA was used to detect the expression levels of IL-1β,IL-6 and TNF-α at different time periods(6,12 and 24 h).The re-sults showed as follows:After 6 h of JEV infection,the mRNA levels of TLR3,TLR7,TRIF and MyD88 genes were significantly up-regulated(P＜0.05),and the mRNA levels of TLR8 genes were down-regulated(P＜0.05).Western blot results showed that the protein expressions of TLR3,TLR7,TRIF and MyD88 were significantly up-regulated when JEV infected testicular stromal cells for 6 h(P＜0.05),which was consistent with the corresponding mRNA transcription levels.There was no significant change in TLR8 protein expression.ELISA results showed that 6 h after JEV infection of testicular stromal cells,IL-6 was significantly increased(P＜0.01),and the expressions of IL-1β and TNF-α were not changed.TLR3,TLR7,TLR8,TRIF and MyD88 were si-lenced by siRNA,and the silenced cells were inoculated with JEV for 6 h,and IL-6 expression lev-els were detected by ELISA.The results showed that silenced TLR3,TLR7,TLR8,TRIF and MyD88 could significantly reduce the increase of IL-6 secretion induced by JEV infection(P＜0.05).These results indicated that JEV could induce the expression of inflammatory factor IL-6 by activating TLR3,TLR7 and TLR8 signaling pathway after infection of testicular stromal cells.This study provides a reference for further elucidating the mechanism of reproductive disorders caused by JEV infection.

Objective:To construct a nomogram for predicting unfavorable prognosis at 6 months after moderate and severe traumatic brain injury (msTBI) and validate its predictive effectiveness.Methods:A retrospective cohort study was conducted to analyze the clinical data of 387 patients with msTBI who were admitted to 904th Hospital of the Joint Logistic Support Force of PLA from January 2020 to December 2022, including 265 males and 122 females, aged 6-97 years [58(47, 68)years]. According to the Glasgow outcome scale (GOS) score at 6 months after injury, the patients were divided into favorable prognosis group (GOS 4-5 points, n=201) and unfavorable prognosis group (GOS 1-3 points, n=186). The clinical characteristics, imaging manifestations, and laboratory test results of the two groups on admission were recorded. Univariate analysis was applied to evaluate the correlation between the aforementioned indicators and the unfavorable prognosis of the msTBI patients at 6 months after injury. Receiver operating characteristic (ROC) curves of single variable and the correlation heatmap among continuous variables were plotted. Lasso regression was used to select variables and multivariate Logistic regression analysis was used to determine independent predictive factors so as to construct Logistic regression equation and plot the nomogram. The internal verification was carried out by means of random and non-random split of data. In random split, the data were divided randomly with a ratio of 6∶4 into training group ( n=232) and verification group ( n=155). In non-random split, the patients admitted from January 2020 to December 2021 were assigned to the training group ( n=260), while those admitted from January 2022 to December 2022 to the verification group ( n=127). Area under the curve (AUC) was used to evaluate the predictive ability of the model in the training group and verification group, calibration curve and Hosmer-Lemeshow (H-L) test to evaluate its goodness of fit, and decision curve analysis (DCA) to evaluate its clinical applicability. The influence of inclusion of neutrophil-to-lymphocyte ratio (NLR) model on the warning effectiveness of poor prognosis was analyzed in comparison with the model without inclusion of NLR. Results:Univariate analysis showed that there was a certain correlation between age, length of hospital stay, Glasgow coma scale (GCS), American Society of Anesthesiologists Physical Status (ASA-PS) classification, Injury severity score (ISS), prehospital tracheal intubation, hypotension, hypoxia, pupillary responsiveness, midline shift, basilar cisterna status, traumatic subarachnoid hemorrhage (tSAH), D-Dimer, prothrombin time activity (PTA), glucose, hemoglobin, K +, Cl -, Ca 2+, HCO -, creatinine, albumin, lactic acid, platelet, lymphocyte, systemic immune-inflammation index (SII), NLR, lymphocyte-to-monocyte ratio (LMR) and unfavorable prognosis of msTBI patients at 6 months after injury ( P<0.05 or 0.01). The ROC curve of single variable showed that GCS (AUC=0.82), ISS (AUC=0.81), pupillary responsiveness (AUC=0.76), basal cistern status (AUC=0.73) and NLR (AUC=0.73) had good predictive validity. The results of the correlation heatmap showed that there was a significant correlation and collinearity among the continuous variables, while no collinearity was found between ISS and NLR. Fourteen potential predictors selected by Lasso regression were included in multivariate Logistic regression analysis and its results showed that age ( OR=0.86, 95% CI 1.38, 5.19), GCS 6-8 points ( OR=3.13, 95% CI 1.06, 9.27), GCS 3-5 points ( OR=12.36, 95% CI 2.81, 54.27), ISS ( OR=3.68, 95% CI 1.38, 9.80), pupillary responsiveness ( OR=2.45, 95% CI 0.85, 7.07), and NLR ( OR=2.62, 95% CI 1.52, 4.51) were identified as the independent risk factors for unfavorable prognosis of msTBI patients at 6 months after injury ( P<0.05 or 0.01). The multivariate Logistic regression equation was Logit [P/(1-P)]=0.066×"age"+ 1.474×"GCS 6-8"+2.357×"GCS 3-5"+0.066×"ISS"+0.965×"absence of pupillary light reflex"+0.194×"NLR"-10.704. In the internal verification of random split of data, the AUC value of the model was 0.93 (95% CI 0.89, 0.96) in the training group and 0.93 (95% CI 0.89, 0.97) in the verification group. In the internal verification of non-random split, the AUC value was 0.94 (95% CI 0.91, 0.97) in the training group and 0.93 (95% CI 0.89, 0.97) in the verification group. The calibration curve and H-L test showed that the model had good calibration ability ( P>0.5). The results of DCA showed that the application of the nomogram would increase the net benefit of the patients (risk threshold probability of 0.0-0.8). Compared with the conventional model (AUC=0.90), inclusion of NLR model (AUC=0.93) enhanced the warning effectiveness. Conclusions:Age, GCS, ISS, pupillary responsiveness and NLR are independent risk factors affecting unfavorable prognosis in msTBI patients at 6 months after injury, based on which the nomogram constructed can better predict the clinical outcome of msTBI patients.

Objective:To investigate the value of three-dimensional (3D) T 1WI structural images using different acceleration methods including parallel acquisition technique, joint compressed sensing (uCS) technique, and artificial intelligence-assisted compressed sensing (ACS) technique for brain region segmentation. Methods:In this cross-sectional study, fifty patients (female: n=25, age range: 13 to 87 years old) at Corning Hospital of Ningbo University from July to September 2023 were prospectively and consecutively collected. All the subjects underwent brain MRI. Six groups of 3D T 1WI structural images were obtained using different acceleration technique and parameters, including 3D T 1WI without acceleration factor (3D-T 1WI group), 3D T 1WI with parallel acquisition technique with acceleration factor 3 (3D-T 1WI-PI-3 group), 3D T 1WI with uCS technique with acceleration factor 4.5 and 6.9 (3D-T 1WI-uCS-4.5 group, 3D-T 1WI-uCS-6.9 group), 3D T 1WI by ACS technique with acceleration factors of 3 and 5 (3D-T 1WI-ACS-3 group, 3D-T 1WI-ACS-5 group). T 2WI fluid-attenuated inversion recovery (FLAIR) images were also acquired. Subjective scores (cerebral grey matter and white matter clarity scores, clarity scores of cerebral white matter degeneration lesions in relation to the surrounding white matter, and Gibbs artifact scores) and objective metrics [signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), cerebrospinal fluid signal homogeneity, peak signal-to-noise ratio (PSNR), structural similarity index (SSIM), and natural image quality evaluator (NIQE)] were used to evaluate image quality in different groups. Totally 109 brain regions were segmented and volumes were measured using the uAI Research Portal image analysis tool. Kappa or intraclass correlation coefficient ( ICC) was used to evaluate the agreement of subjective and objective evaluation indexes between the 3D-T 1WI-PI-3 group, 3D-T 1WI-uCS-4.5 group, 3D-T 1WI-uCS-6.9 group, 3D-T 1WI-ACS-3 group, 3D-T 1WI-ACS-5 group, and 3D-T 1WI group. Kappa or ICC value>0.70 was considered as good agreement. Results:The acquisition time for the 3D-T 1WI group, 3D-T 1WI-PI-3 group, 3D-T 1WI-uCS-4.5 group, 3D-T 1WI-uCS-6.9 group, 3D-T 1WI-ACS-3 group, and 3D-T 1WI-ACS-5 group were 527, 204, 169, 95, 133, 90 s, respectively. Subjective evaluation showed that the 3D-T 1WI-uCS-4.5, 3D-T 1WI-ACS-3, and 3D-T 1WI-ACS-5 groups had excellent agreement with the 3D-T 1WI group in terms of the distribution of cases of cerebral grey matter and white matter clarity scores, respectively (all Kappa value=1.000); The distribution of cases of clarity score of cerebral white matter lesions and surrounding white matter in the 3D-T 1WI-PI-3 group, 3D-T 1WI-uCS-4.5 group, and 3D-T 1WI-ACS-3 group were in good agreement with that of the 3D-T 1WI group ( Kappa values of 0.775, 0.701, and 0.777, respectively); the distribution of the number of cases of the Gibbs artifact score of the 3D-T 1WI-uCS-4.5, 3D-T 1WI-ACS-3, and 3D-T 1WI-ACS-5 groups was in good agreement with the 3D-T 1WI group (all Kappa value=1.000). Objective evaluation showed the CNR of the images in the 3D-T 1WI-PI-3, 3D-T 1WI-uCS-4.5, and 3D-T 1WI-uCS-6.9 groups were in good agreement with those of the 3D-T 1WI group ( ICC of 0.720, 0.759, and 0.752, respectively); PSNR and SSIM were in good agreement among the 3D-T 1WI-PI-3 group, 3D-T 1WI-uCS-4.5 group, 3D-T 1WI-uCS-6.9 group, 3D-T 1WI-ACS-3 group, and 3D-T 1WI-ACS-5 group (PSNR: ICC=0.854; SSIM: ICC=0.851). NIQE of 3D-T 1WI-PI-3 group, 3D-T 1WI-uCS-4.5 group, and 3D-T 1WI-ACS-3 group images were in good agreement with the 3D-T 1WI group ( ICC value of 0.866, 0.727, 0.753, respectively). The ICC values of the volume of each segmented brain region among the 3D-T 1WI-PI-3, 3D-T 1WI-uCS-4.5, 3D-T 1WI-uCS-6.9, 3D-T 1WI-ACS-3, 3D-T 1WI-ACS-5 group and the 3D-T 1WI group images showed decreased in order (all ICC≥0.62). Conclusions:The uCS and ACS techniques used in 3D-T 1WI show high agreement with 3D-T 1WI in terms of brain segmentation. The application of these accelerating techniques can significantly shorten the acquisition time with obtaining images with good image quality, displaying great value.

Objective:To explore the clinical features and genetic etiology of a patient with adult-onset globoid cell leukodystrophy/Krabbe disease (KD).Methods:A patient who was admitted to the Tongji Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology on February 15, 2022 due to exacerbation of right leg weakness for over 4 years was selected as the study subject. Clinical data and results of medical imaging and genetic analysis were analyzed. Candidate variants were verified by family analysis.Results:The patient, a 36-year-old woman, had spasmodic gait as the primary presentation. Cranial magnetic resonance imaging (MRI) revealed symmetrical abnormalities in the bilateral corticospinal tracts, and the activity of β-galactocerebrosidase (GALC) in her white blood cells was significantly decreased. The patient was found to harbor compound heterozygous variants of the GALC gene, namely c.461C>A (p.Pro154His) and c. 1901T>C (p.Leu634Ser). Her mother, sister and nephew were heterozygous carriers of the c. 461C>A (p.Pro154His) variant, whilst her father was heterozygous for the c.1901T>C (p.Leu634Ser) variant. Conclusion:The patient was ultimately diagnosed with adult-onset KD, for which the compound heterozygous variants of the GALC gene may be accountable.