Objective: To understand the allocation and use of safety seats for preschool children and explore its related factors, so as to provide a scientific reference for promoting the usage of safety seats. Methods: A stratified random cluster sampling method was used to select 3 143 parents of preschool children aged 3 to 6 from six kindergartens in Shunyi District, Beijing from January 3 to 10, 2022. An online questionnaire survey was conducted to collect and evaluate the equipment and use of child safety seats in different characteristics of preschool children, as well as their scores of health beliefs. Multiple factor Logistic regression analysis was used to investigated the related factors of safety seat configuration and use. Results: The equipping rate and usage rate of safety seats for preschool children were 66.56% and 58.45%, respectively. The proportion of equipped and used safety seats for preschool children in core families (69.52%, 62.23%) were higher than that in large families (64.35%, 55.62%), only child families ( 72.39 %, 64.87%) were higher than non only child families (61.49%, 52.86%), and urban families (71.63%, 63.04%) were higher than rural families (52.31%, 45.51%) ( χ 2=9.23, 13.86; 41.72, 46.44; 101.96 ,76.97,all P <0.05) . As the educational level of parents ( χ 2 trend =154.23,98.76) and annual income of the family ( χ 2 trend =155.78,127.69) rised, the reporting rates of the equipped and used child safety seats in the family also increased(all P <0.05 ). There were statistically significant differences in the scores of different dimensions of health beliefs for the provision ( t =-20.22-18.16) and use ( t =24.32-24.17) of safety seats for preschool children(all P <0.05). After adjusting for child sex, child age, family annual income, parental education level, family type, whether the child was an only child, and place of residence，multivariate Logistic regression analysis showed that preschool children with higher perceived susceptibility score( OR =1.11, 1.08), higher self efficacy score( OR =1.23, 1.33), and higher suggestive factors score( OR =1.08, 1.12) were more likely to have and use safety seats in their families, while preschool children with higher perceived impairments score( OR =0.82, 0.80) were less likely to have and use safety seats in their families (all P <0.05). Conclusions The installation rate of child safety seats needs to be improved, and there is also a certain gap in their use after installation. Parents of preschool children should improve susceptibility and self efficacy to safety seat equipment and use, and perceptual barriers should be reduced.

ObjectiveTo investigate the role of runt-related transcription factor 3 （RUNX3） in transforming growth factor-β₁ （TGF-β₁）-induced activation of mouse primary pulmonary fibroblasts （PFs）， and its effects on fibroblast activation protein （FAP） expression， cell proliferation， and collagen synthesis. MethodsPFs were isolated from C57BL/6 mice and cultured. A RUNX3 knockdown model was established using small interfering RNA （siRNA）. Cells were assigned to the control group （Control）， TGF-β₁-treated group （TGF-β₁）， negative control group （TGF-β₁+siRNA-NC）， and RUNX3-silenced group （TGF-β₁+si-RUNX3）. In addition， a RUNX3 overexpression rescue experiment was performed based on TGF-β₁ stimulation. Protein and mRNA levels of RUNX3， FAP， and typeⅠcollagen （COL1A1） were measured by Western blot and reverse transcription quantitative real-time PCR （RT-qPCR）. Cell proliferation was assessed using CCK-8 and EdU assays. Co-expression of COL1A1 and FAP was examined by double immunofluorescence staining. ResultsCompared with the Control group， RUNX3， FAP， and COL1A1 expression levels were upregulated in PFs in the TGF-β₁ group （P<0.01）. The CCK-8 assay showed that the absorbance value was reduced in the RUNX3 knockdown group compared with the negative control group （P<0.01）. Consistently， the EdU assay demonstrated a lower proportion of EdU-positive cells in the RUNX3 knockdown group than in the negative control group （P<0.01）. Immunofluorescence double staining revealed decreased fluorescence intensities of COL1A1 and FAP in the RUNX3 knockdown group relative to the negative control. Under RUNX3 overexpression conditions， these fluorescence signals exhibited a partial rebound （P<0.01）. ConclusionRUNX3 in TGF-β1-induced PFs may promote cell proliferation and collagen synthesis by positively regulating FAP expression. Targeting the RUNX3/FAP axis may represent a potential therapeutic strategy for pulmonary fibrosis.

Objective: To investigate the effect of targeted silencing of DNA methyltransferase 3A(DNMT3A) on collagen deposition, proliferation and migration activity of mouse lung fibroblasts(PFs). Methods: In order to ensure the proliferation and migration activity of primary fibroblasts, the lung tissues of neonatal C57 suckling mice were taken, PFs were extracted after being sheared, and the morphology was observed and identified under the microscope. PFs cells were activated by 5 ng/ml TGF-β1for 24 h after cell attachment, and DNMT3A silencing model was constructed by small interfering RNA; The experiment was divided into control group, TGF-β1group, TGF-β1+ siRNA-NC group and TGF-β1+ siRNA-DNMT3A group. The protein expressions of DNMT3A, α-smooth muscle actin(α-SMA) and Collagen Ⅰ were detected by Western blot; Real time quantitative reverse transcription polymerase chain reaction(RT-qPCR) was used to detect the mRNA expression changes ofDNMT3A,α-SMAandCollagenⅠ. The proliferation ability of PFs was detected by CCK-8 and EdU staining; the migration ability of PFs was detected by scratch test and Transwell migration test. Results: Compared with the control group, TGF-β1induced the increase of DNMT3A in the activated PFs cell group(P<0.01), the protein and mRNA levels of fibrosis and proliferation related indicators α-SMA and Collagen Ⅰ also increased(allP<0.05), and the proliferation and migration ability of PFs increased(allP<0.000 1). Compared with the siRNA-NC group, the protein expression levels of DNMT3A(P<0.000 1) and related indicators α-SMA(P<0.01) and Collagen Ⅰ(P<0.01) significantly decreased in the DNMT3A silencing group by Western blot, and the mRNA levels ofDNMT3A,α-SMAandCollagenⅠby RT-qPCR also decreased(allP<0.001), and the proliferation(P<0.01) and migration ability(P<0.05) of PFs cells decreased compared with the control group. Conclusion Silencing DNMT3A can inhibit the deposition of collagen and the proliferation of PFs. DNMT3A can promote the proliferation and migration of PFs, and then promote the activation of PFs and the development of pulmonary fibrosis. This process may be regulated by DNA methylation modification.

Objective : To clarify the intrinsic link between a high-fat diet(HFD) and the pathological progression and prognosis of spinal cord injury(SCI) while preliminarily exploring the potential underlying mechanisms. Methods: SCI models were established in mice that were fed either a regular diet(RD) or HFD, with injury inflicted specifically on the T9-T12segments. Hematoxylin-Eosin(HE) staining, Masson staining, and Nissl staining were used to observe the local histological changes in SCI tissues. The basso, beattie, and bresnahan(BBB) score and footprint analysis were used to evaluate and compare hindlimb functional recovery after SCI in both RD and HFD mice.In vitroexperiments were conducted to identify key fatty acids in the HFD that exacerbate neuronal damage, whilein vivoexperiments assessed the effects of 2-bromopalmitate(2-BP), a palmitic acid inhibitor, on HFD-fed mice with SCI. Results : Compared to RD-fed mice, HFD-fed mice exhibited significantly larger lesion areas, more severe neuronal damage, and poorer hindlimb functional recovery after SCI. Palmitic acid was identified as the key fatty acid aggravating neuronal damage. Further more, inhibition of palmitoylation, mediated by palmitic acid, enhanced neuronal survival, promoted tissue repair, and improved hindlimb functional recovery in HFD-fed mice post-SCI. Conclusion HFD exacerbates pathological damage following SCI in mice through palmitic acid, impairing recovery. Palmitic acid-mediated palmitoylation is likely the main mechanism underlying this effect.

Hypothermia is a clinical syndrome characterized by core body temperature<35℃,caused by significant heat loss from body surface in cold environment.As a systemic cold injury,it can be lethal if treatment is delayed.Emergent diagnosis and treatment of hypothermia are expected to improve the prognosis of patients.In 2005,the U.S.Army Research Institute of Environmental Medicine(USARIEM)issued guidelines for the prevention and management of cold injuries,but there has been no corresponding standard in China.Therefore,Emergency Branch of Chinese Medical Rescue Association,Emergency Medical Equipment Society of China Association of Medical Equipment,Integrated Rehabilitation Medical Branch of Chinese Medical Rescue Association,and Pre-Hospital Emergency Care Working Committee of Chinese Aging Well Association jointly developed the Chinese Expert Consensus on Emergent Treatment of Hypothermia(2025 edition).The consensus covers the pathophysiology,etiology and epidemiology,diagnosis and severity grading,prehospital treatment,and in-hospital treatment of hypothermia,including 15 recommendations in total,aiming to provide guidance for the relevant clinical rescue work.

Objective:The haplotypes of Killer cell immunoglobulin-like receptors (KIR) can be divided into centromeric and telomeric ones. As the terminal gene at the centromeric end, KIR3DP1 plays an important role in stabilizing the haplotype structure. This study aimed to analyze the distribution of KIR3DP1 gene haplotypes among Han Chinese population in Zhejiang in order provide a basis for further analyzing the role of KIR3DP1 in the KIR haplotypes. Methods:A total of 166 unrelated blood donors from Zhejiang were collected (Blood donation period: March 2020 to August 2020), and genotyping was performed by next-generation sequencing based on exon capture. The copy number and allelic frequency of the KIR3DP1 gene and the distribution of centromeric haplotypes were statistically analyzed. This study was approved by the Medical Ethics Committee of Zhejiang Blood Center (Ethics No.: 2023-001). Results:The KIR3DP1 gene was positive for all individuals but with different copy numbers. Among these, 4 cases (2.4%) had only 1 copy, 156 cases (94.0%) had 2 copies, and 6 cases (3.6%) had 3 copies. A total of 10 KIR3DP1 alleles were found in the population, which could be classified into the KIR3DP1*001-L type, KIR3DP1*003-L type, and KIR3DP1 full deletion type. The KIR3DP1*003-L type allele was linked to the Cen- A01 and Cen- B01 types, and the KIR3DP1*001-L type allele and the KIR3DP1 deletion type were only present in the Cen- B02 type haplotype. Conclusion:This study has derived a high-resolution distribution map of the KIR3DP1 gene in the Han population from Zhejiang, and found that the KIR3DP1 alleles showed different linkage with the centromeric haplotypes, which has provided a basis for further studying the role of KIR3DP1 in genetic immunity.

Objective: To explore the mechanism of hesperidin on hindlimb motor function in mice with spinal cord injury(SCI). Methods: Oxygen glucose deprivation(OGD) was used to induce SCI in PC12 neuronal cells.CCK-8 assay determined the optimal hesperidin concentration(0,10,20,30,40,50,60 mmol/L) for subsequent experiments after 24 h treatment. PC12 cells were divided into Control group,OGD group and Hesperidin(30 mmol/L) + OGD groups. Hoechst staining assessed apoptosis; Western blot(WB),qPCR and immunofluorescence(IF) detected the expression of i NOS,CD206 and TNF-α. Flow cytometry measured apoptosis rates,and WB examined the impact of hesperidin on PI3K,p-PI3K,AKT,p-AKT,and NF-κB expression. Forty-five healthy male C57BL/6 mice were randomly divided into Sham group,Sci group and Hesperidin groups. Sham and Sci groups received 0. 1 mL PBS,while the Hesperidin group received 0. 1 mL hesperidin. After modeling,i NOS,CD206,and TNF-α expression levels,spinal cord cavitation,and hindlimb motor function were evaluated. Results: Hesperidin at 10-30 mmol/L for 24 h did not significantly affect PC12 cell activity. Hoechst staining showed reduced apoptosis in PC12 cells after 24 h of 30 mmol/L hesperidin treatment. Compared to the OGD group,Hesperidin + OGD group had decreased mRNA and protein expression of inflammatory factors(i NOS and TNF-α) and increased expression of the anti-in ammatory factor(CD206). Hesperidin treatment inhibited PI3K/AKT/NF-κB pathway activation,with similar results in mouse experiments. Compared to Sci group,Hesperidin group had reduced spinal cord cavitation area and improved hindlimb motor function. Conclusion Hesperidin may improve hindlimb motor function in mice with SCI by downregulating the phosphorylation level of the PI3K/AKT/NF-κB pathway,inhibiting in ammatory factor release,promoting anti-inflammatory factor release,regulating the inflammatory microenvironment after SCI,and suppressing the acute inflammatory response.

Thalidomide(THA)is renowned for its potent anti-inflammatory properties.This study aimed to eluci-date its underlying mechanisms in the context of Crohn's disease(CD)development.Mouse colitis models were established by dextran sulfate sodium(DSS)treatment.Fecal microbiota and metabolites were analyzed by metagenomic sequencing and mass spectrometry,respectively.Antibiotic-treated mice served as models for microbiota depletion and transplantation.The expression of forkhead box P3+(FOXP3+)regulatory T cells(Tregs)was measured by flow cytometry and immunohistochemical assay in colitis model and patient cohort.THA inhibited colitis in DSS-treated mice by altering the gut microbiota profile,with an increased abundance of probiotics Bacteroides fragilis,while pathogenic bacteria were depleted.In addition,THA increased beneficial metabolites bile acids and significantly restored gut barrier function.Transcriptomic profiling revealed that THA inhibited interleukin-17(IL-17),IL-1β and cell cycle signaling.Fecal microbiota transplantation from THA-treated mice to microbiota-depleted mice partly recapitulated the effects of THA.Specifically,increased level of gut commensal B.fragilis was observed,correlated with elevated levels of the microbial metabolite 3alpha-hydroxy-7-oxo-5beta-cholanic acid(7-ketolithocholic acid,7-KA)following THA treatment.This microbial metabolite may stable FOXP3 expression by targeting the receptor FMR1 autosomal homolog 1(FXR1)to inhibit auto-phagy.An interaction between FOXP3 and FXR1 was identified,with binding regions localized to the FOXP3 domain(aa238-335)and the FXR1 domain(aa82-222),respectively.Conclusively,THA modu-lates the gut microbiota and metabolite profiles towards a more beneficial composition,enhances gut barrier function,promotes the differentiation of FOXP3+Tregs and curbs pro-inflammatory pathways.

OBJECTIVE: The haplotypes of Killer cell immunoglobulin-like receptors (KIR) can be divided into centromeric and telomeric ones. As the terminal gene at the centromeric end, KIR3DP1 plays an important role in stabilizing the haplotype structure. This study aimed to analyze the distribution of KIR3DP1 gene haplotypes among Han Chinese population in Zhejiang in order provide a basis for further analyzing the role of KIR3DP1 in the KIR haplotypes. METHODS: A total of 166 unrelated blood donors from Zhejiang were collected (Blood donation period: March 2020 to August 2020), and genotyping was performed by next-generation sequencing based on exon capture. The copy number and allelic frequency of the KIR3DP1 gene and the distribution of centromeric haplotypes were statistically analyzed. This study was approved by the Medical Ethics Committee of Zhejiang Blood Center (Ethics No.: 2023-001). RESULTS: The KIR3DP1 gene was positive for all individuals but with different copy numbers. Among these, 4 cases (2.4%) had only 1 copy, 156 cases (94.0%) had 2 copies, and 6 cases (3.6%) had 3 copies. A total of 10 KIR3DP1 alleles were found in the population, which could be classified into the KIR3DP1*001-L type, KIR3DP1*003-L type, and KIR3DP1 full deletion type. The KIR3DP1*003 L type allele was linked to the Cen-A01 and Cen-B01 types, and the KIR3DP1*001*L type allele and the KIR3DP1 deletion type were only present in the Cen-B02 type haplotype. CONCLUSION This study has derived a high-resolution distribution map of the KIR3DP1 gene in the Han population from Zhejiang, and found that the KIR3DP1 alleles showed different linkage with the centromeric haplotypes, which has provided a basis for further studying the role of KIR3DP1 in genetic immunity.
Adult ; Female ; Humans ; Male ; Alleles ; China/ethnology* ; Gene Frequency ; Genotype ; Haplotypes/genetics* ; High-Throughput Nucleotide Sequencing/methods* ; East Asian People/genetics* ; Receptors, KIR/genetics*

Protein tyrosine phosphatase nonreceptor type 2 (PTPN2) is a promising target for sensitizing solid tumors to immune checkpoint blockades. However, the highly polar active sites of PTPN2 hinder drug discovery efforts. Leveraging small interfering RNA (siRNA) technology, we developed a novel glutathione-responsive nano-platform HPssPT (HA/PEIss@siPtpn2) to silence PTPN2 and enhance immunotherapy efficacy in hepatocellular carcinoma (HCC). HPssPT showed potent transfection and favorable safety profiles. PTPN2 deficiency induced by HPssPT amplified the interferon γ signaling in HCC cells by increasing the phosphorylation of Janus-activated kinase 1 and signal transducer and activator of transcription 1, resulting in enhanced antigen presentation and T cell activation. The nano-platform was also able to promote the M1-like polarization of macrophages in vitro. The unique tropism of HPssPT towards tumor-associated macrophages, facilitated by hyaluronic acid coating and CD44 receptor targeting, allowed for simultaneous reprogramming of both tumor cells and tumor-associated macrophages, thereby synergistically reshaping tumor microenvironment to an immunostimulatory state. In HCC, colorectal cancer, and melanoma animal models, HPssPT monotherapy provoked robust antitumor immunity, thereby sensitizing tumors to PD-1 blockade, which provided new inspiration for siRNA-based drug discovery and tumor immunotherapy.