Objective:To analyze the protective effect of a respiratory syncytial virus (RSV) vaccine based on bacterial outer membrane vesicle (OMV) in mice.Methods:The pre-fusion protein (preF) of RSV was linked to the surface of OMV through the transmembrane protein cytolysin A (ClyA) to form the nanovaccine OMV-preF. The morphological characteristics of OMV and OMV-preF were observed under a transmission electron microscope. OMV-preF was intramuscularly injected into BALB/c mice and the elicited humoral and cellular immune responses were evaluated. The protective effect of OMV-preF was assessed by challenging the immunized mice with RSV Long strain. One-way analysis of variance and Tukey test were used for statistical analysis.Results:The results showed that preF was stably expressed in OMV, and both OMV-preF and OMV exhibited a double-layer vesicle structures under the microscope. OMV-preF could significantly activate the cellular and humoral immune responses in mice, causing a significant increase in CD8 + T cells and CD19 + B cells as well as a significant increase in the serum level of specific IgG. The neutralizing antibodies produced in the immunized mice could significantly inhibit the replication of RSV Long strain in vivo. Conclusions:The nanovaccine OMV-preF can induce high-level humoral and cellular immune responses, and the antibodies produced following immunization can effectively inhibit viral replication. This study provides a new strategy for RSV subunit vaccines.

OBJECTIVE: This study investigates the efficacy and mechanisms of Qingjie Fuzheng Granules (QFG) in inhibiting colitis-associated colorectal cancer (CAC) development via RNA sequencing (RNA-seq) and 16S ribosomal RNA (rRNA) correlation analysis. METHODS: CAC was induced in BALB/c mice using azoxymethane (AOM) and dextran sulfate sodium (DSS), and QFG was administered orally to the treatment group. The effects of QFG on CAC were evaluated using disease index, histology, and serum T-cell ratios. RNA-seq and 16S rRNA analysis assessed the transcriptome and microbiome change. Key pharmacodynamic pathways were identified by integrating these data and confirmed via Western blotting and immunofluorescence. The link between microbiota and CAC-related markers was explored using linear discriminant analysis effect size and Spearman correlation analysis. RESULTS: Long-term treatment with QFG prevented AOM/DSS-induced CAC formation, reduced levels of interleukin (IL)-1β, tumor necrosis factor-alpha (TNF-α), IL-6, and interferon γ (IFN-γ), and increased CD3⁺ and CD4⁺/CD8⁺ T cells ratio, without causing hepatic or renal toxicity. A 16S rRNA analysis revealed that QFG rebalanced the Firmicutes/Bacteroidetes ratio and mitigated AOM/DSS-induced microbiota disturbances. Transcriptomics and Western blotting analysis identified the nucleotide-binding oligomerization domain-containing protein 2 (NOD2)/nuclear factor kappa-B (NF-κB) pathway as key for QFG's treatment against CAC. Furthermore, QFG decreased the abundance of Bacilli, Bacillales, Staphylococcaceae, Staphylococcus, Lactobacillales, Aerococcus, Alloprevotella, and Akkermansia, while increasing Clostridiales, Lachnospiraceae, Lachnospiraceae_NK4A136_group, Ruminococcaceae, and Muribaculaceae, which were highly correlated with CAC-related markers or NOD2/NF-κB pathway. CONCLUSION By mapping the relationships between CAC, immune responses, microbiota, and key pathways, this study clarifies the mechanism of QFG in inhibiting CAC, highlighting its potential for clinical use as preventive therapy.

Objective To explore the diagnostic value of deep learning image reconstruction(DLIR)com-bined with low-dose chest computed tomography(CT)with axial scan in the diagnosis of mycoplasma pneumoniae pneumonia(MPP)in children,and to provide reference for clinical practice.Methods 160 cases MPP children from February 2024 to June 2025 were selected as study subjects,and low-dose chest CT with axial scan was performed on all patients.DLIR and conventional adaptive iterative reconstruction-V(ASIR-V)were used for image reconstruction.The objective image quality[background noise(SD),signal-to-noise ratio(SNR),and contrast-to-noise ratio(CNR)],subjective image quality,and CT sign detection rate were compared,and the consistency of DLIR and ASIR-V in the diagnosis of MPP severity and clinical diagnosis was compared.Results As the intensity of DLIR and the weight of ASIR increasd,SD gradually decreased,while SNR and CNR gradually increased.The high-strength DLIR(DLIR-H)SD was lower than that of ASIR with a blending level of 80%(ASIR-V80%).The SNR and CNR were higher than those of ASIR-V80%,showing statistical significance(P<0.05).Ridit test showed that DLIR-H had the best subjective image quality score under different DLIR intensities,and ASIR-V80%had the best subjective image quality score under different ASIR weights.Furthermore,the subjective image quality score of DLIR-H was higher that of ASIR-V80%,and the differences were statistically significant(P<0.05).Using DLIR-H,the detection rates of air bronchogram,pulmonary consolidation,and interstitial infiltration(69.38%,86.88%,20.63%,respectively)were higher than those using ASIR-V80%(50.00%,71.88%,7.50%,respectively),and the differences were statistically significant(P<0.05).Consistency analysis showed that the Kappa value between the diagnostic results of MPP severity using DLIR-H and clinical diagnosis was 0.856(95%CI:0.771～0.996),while that between the diagnostic results of MPP severity using ASIR-V80%and clinical diagnosis was 0.498(95%CI:0.346～0.650).ROC analysis showed that the area under the curve(AUC)for diagnosing MPP severity was 0.925(95%CI:0.872～0.960)for DLIR-H and 0.729(95%CI:0.653～0.796)for ASIR-V80%,and the diagnostic value of DLIR-H was superior to that of ASIR-V80%(Z=3.952,P<0.001).Conclusion DLIR can effectively improve image quality.DLIR-H combined with low-dose chest CT with axial scan has high diagnostic value for the severity of MPP,and can serve as a feasible solution for clinical diagnosis of MPP severity and reducing radiation dose.

Objective To explore the diagnostic value of deep learning image reconstruction(DLIR)com-bined with low-dose chest computed tomography(CT)with axial scan in the diagnosis of mycoplasma pneumoniae pneumonia(MPP)in children,and to provide reference for clinical practice.Methods 160 cases MPP children from February 2024 to June 2025 were selected as study subjects,and low-dose chest CT with axial scan was performed on all patients.DLIR and conventional adaptive iterative reconstruction-V(ASIR-V)were used for image reconstruction.The objective image quality[background noise(SD),signal-to-noise ratio(SNR),and contrast-to-noise ratio(CNR)],subjective image quality,and CT sign detection rate were compared,and the consistency of DLIR and ASIR-V in the diagnosis of MPP severity and clinical diagnosis was compared.Results As the intensity of DLIR and the weight of ASIR increasd,SD gradually decreased,while SNR and CNR gradually increased.The high-strength DLIR(DLIR-H)SD was lower than that of ASIR with a blending level of 80%(ASIR-V80%).The SNR and CNR were higher than those of ASIR-V80%,showing statistical significance(P<0.05).Ridit test showed that DLIR-H had the best subjective image quality score under different DLIR intensities,and ASIR-V80%had the best subjective image quality score under different ASIR weights.Furthermore,the subjective image quality score of DLIR-H was higher that of ASIR-V80%,and the differences were statistically significant(P<0.05).Using DLIR-H,the detection rates of air bronchogram,pulmonary consolidation,and interstitial infiltration(69.38%,86.88%,20.63%,respectively)were higher than those using ASIR-V80%(50.00%,71.88%,7.50%,respectively),and the differences were statistically significant(P<0.05).Consistency analysis showed that the Kappa value between the diagnostic results of MPP severity using DLIR-H and clinical diagnosis was 0.856(95%CI:0.771～0.996),while that between the diagnostic results of MPP severity using ASIR-V80%and clinical diagnosis was 0.498(95%CI:0.346～0.650).ROC analysis showed that the area under the curve(AUC)for diagnosing MPP severity was 0.925(95%CI:0.872～0.960)for DLIR-H and 0.729(95%CI:0.653～0.796)for ASIR-V80%,and the diagnostic value of DLIR-H was superior to that of ASIR-V80%(Z=3.952,P<0.001).Conclusion DLIR can effectively improve image quality.DLIR-H combined with low-dose chest CT with axial scan has high diagnostic value for the severity of MPP,and can serve as a feasible solution for clinical diagnosis of MPP severity and reducing radiation dose.

Objective:To analyze the protective effect of a respiratory syncytial virus (RSV) vaccine based on bacterial outer membrane vesicle (OMV) in mice.Methods:The pre-fusion protein (preF) of RSV was linked to the surface of OMV through the transmembrane protein cytolysin A (ClyA) to form the nanovaccine OMV-preF. The morphological characteristics of OMV and OMV-preF were observed under a transmission electron microscope. OMV-preF was intramuscularly injected into BALB/c mice and the elicited humoral and cellular immune responses were evaluated. The protective effect of OMV-preF was assessed by challenging the immunized mice with RSV Long strain. One-way analysis of variance and Tukey test were used for statistical analysis.Results:The results showed that preF was stably expressed in OMV, and both OMV-preF and OMV exhibited a double-layer vesicle structures under the microscope. OMV-preF could significantly activate the cellular and humoral immune responses in mice, causing a significant increase in CD8 + T cells and CD19 + B cells as well as a significant increase in the serum level of specific IgG. The neutralizing antibodies produced in the immunized mice could significantly inhibit the replication of RSV Long strain in vivo. Conclusions:The nanovaccine OMV-preF can induce high-level humoral and cellular immune responses, and the antibodies produced following immunization can effectively inhibit viral replication. This study provides a new strategy for RSV subunit vaccines.

Background Organic phosphate flame retardants are emerging environmental pollutants. While there have been multiple toxicities reported following organic phosphate flame retardants exposure, few studies focus on their potential developmental toxicities. It is necessary to elucidate these developmental toxicological effects and underlying mechanisms to improve risk assessments and better protect sensitive populations. Objective To evaluate potential developmental toxicities in early chicken embryos following exposure to triphenyl phosphate (TPhP) or cresyl diphenyl phosphate (CDP), to reveal TPhP and CDP’s capabilities to activate peroxisome proliferator-activated receptor γ (PPARγ) in vivo in an established chicken embryo gene reporter system, and to investigate the roles of PPARγ in TPhP/CDP-induced developmental toxicities with lentivirus-mediated in vivo gene silencing. Methods Firstly, diverse doses of TPhP and CDP were injected into the air sacs of fertilized eggs to assess the development of chicken embryos after 6 d of incubation, and an optimal dose was chosen for subsequent experiments. Subsequently, the report gene system was employed to evaluate the intraembryonic activation of PPARγ by TPhP and CDP. Eventually, PPARγ was silenced using lentivirus, and the embryos were co-treated with TPhP and CDP to further disclose the roles of PPARγ in the observed developmental toxicity. Results Following developmental exposure to TPhP or CDP, significantly lower chicken embryo weights (normalized with egg weights) were observed in the 6 d embryos (10, 30 mg·kg⁻¹ TPhP and 3, 10, 30 mg·kg⁻¹ CDP), indicating that both chemicals have general developmental toxicities and CDP is more potent. Additionally, exposure to CDP also resulted in remarkably increased sagittal brain area (normalized to embryo weights) and decreased sagittal eye area (normalized to embryo weights) (P<0.05), suggesting that CDP has specific developmental neurotoxicity and ocular toxicity. The PPARγ reporter gene experiment results revealed that rosiglitazone (positive control), TPhP, and CDP all significantly activated PPARγ relative to control (P<0.05). The potency order was rosiglitazone > CDP > TPhP. The lentivirus microinjection successfully achieved in vivo silencing of PPARγ in developing chicken embryos, and the estimated silencing efficacy was approximately 55% according to the real-time quantitative polymerase chain reaction (qRT-PCR) results. The in vivo silencing of PPARγ effectively alleviated TPhP or CDP-induced decrease of embryo weights (P<0.05), as well as CDP-induced increase of brain areas and decrease of eye areas (P<0.05). Conclusions Both TPhP and CDP can induce general developmental toxicities in early chicken embryos, and CDP is more potent than TPhP. Meanwhile, CDP can induce specific enlarged brain area and decreased eye area. The observed toxicities are associated with in vivo activation of PPARγ.

Objective:To analyze antiviral activity against respiratory syncytial virus (RSV) of interferon (IFN)-α2b and IFN-λ1 on Hep2 cells and human airway epithelial (HAE) cells.Methods:IFN-α2b or IFN-λ1 was incubated with Hep2 cells after RSV infection, and 48 hours later, the cytopathic effect was observed, the viral load was determined using real time/reverse transcription quantitative polymerase chain reaction (RT qPCR), RSV F protein expression was detected using immunofluorescence, and cell survival rate was detected using crystal violet. HAE cells were incubated with IFN-α2b or IFN-λ1 for 24 hours, and then HAE were challenged with RSV. The viral load in the culture supernatant was determined on days 1-7 using RT qPCR, RSV F protein was determined with immunofluorescence and the viral titers in the culture supernatant was detected on day 7 by plaque assay.Results:In Hep2 cells, the CPE of the treatment groups (IFN-α2b and IFN-λ1) was alleviated compared to the virus control group, and the CPE of the high concentration group was lighter than that of the low concentration group. Different concentrations of IFN-α2b and IFN-λ1 could significantly reduce the viral load of RSV ( P<0.001), and the viral load of the high concentration group was significantly lower than that of the low concentration group ( P<0.001). In addition, IFN-α2b and IFN-λ1 could reduce the RSV F protein expression after RSV infection and improve cell survival rate. In HAE cells, IFN-α2b and IFN-λ1 could inhibit RSV virus replication, reduce virus titers ( P<0.001) and reduce RSV F protein expression. Conclusions:IFN-α2b and IFN-λ1 both showed great antiviral activity against RSV in Hep2 and HAE cells, providing data reference for the study of interferon against respiratory viruses.

In recent years, a large number of studies have found that Parkinson's disease (PD) patients often complicate with osteoporotic fracture, which may be related to decreased dopamine transmitters, PD risk gene mutations, mitochondrial dysfunction, vitamin D reduction, anti-PD drug use, autonomic nervous dysfunction and intestinal microflora disorders. Therefore, this paper reviews the current research progress on the pathogenesis of osteoporotic fracture in PD patients to provide theoretical basis for co-morbid treatment of PD and osteoporotic fracture.

Objective:To isolate and culture WU polyomavirus (WUPyV), and to analyze the genome-wide evolutionary patterns, homology and population dynamics.Methods:Real-time quantitative PCR was used to detect the nasopharyngeal aspirate samples of hospitalized children with respiratory tract infection in Beijing Friendship Hospital during 2020 to 2022. Primary human airway epithelial cells cultured at the air-liquid interface were used to isolate and culture WUPyV. Whole genome sequence of the isolated strain was obtained by Sanger sequencing. For phylogenetic and evolutionary dynamics analysis, the whole genome was compared with the published whole genome sequences in GenBank database.Results:The detection rate of WUPyV was 4.7% (31/659) during 2020 to 2022, and a clinical strain BJ0593 of WUPyV type Ⅲc was successfully isolated. The homology of the whole genome and gene fragments of WUPyV was high. The average evolutionary rate of VP2 gene was about 1.256×10 -4 substitution/site every year, and the population dynamics of WUPyV tended to be flat in the last decade. Conclusions:This study successfully isolated a clinical WUPyV type Ⅲ strain for the first time, which provided the basis for further investigation on the molecular evolution and pathogenicity of WUPyV.

Objective:To investigate the epidemiological characteristics of human bocavirus 1 (HBoV1) and to analyze the genetic variation.Methods:A total of 2 848 nasopharyngeal aspirate (NPAs) specimens were collected from hospitalized children with acute respiratory tract infections (ARTI) in Beijing Friendship Hospital from April 2017 to March 2019, and HBoV1 was detected by quantitative real-time PCR. Epidemiological analysis was carried out based on the clinical information of the patients. The nested PCR method was used to amplify the NP1 and VP1 genes of HBoV1 for homology analysis. Maximum clade credibility tree (MCC tree) and genetic polymorphism map were constructed to analyze the time evolution of HBoV1 VP1.Results:HBoV1 was detected in 90(3.16%) of 2 848 NPAs, most (93.33%, 84/90) HBoV1-positive cases were among children <5 years of age. HBoV1 could be detected throughout the year with a higher prevalence 7.23% (18/249) in October. Of the 90 HBoV1-infected cases, the main clinical symptoms were fever and cough, 44(48.89%) were co-infected with other respiratory viruses; 55 NP1 sequences and 47 VP1 sequences were obtained by nested PCR amplification, phylogenetic analysis showed that the nucleotide homology was 98.9%~-100% and 99.1%~-100%, respectively. MCC tree showed that the HBoV1 VP1 gene sequence obtained in this study appeared in two adjacent clades, the gene evolution was stable.Conclusions:HBoV1 is one of the common viruses that cause respiratory infection among children in Beijing. HBoV1 genetic evolution is relatively stable, but it still needs to be monitored continuously.