Objective: To understand classroom environmental conditions and their association with spinal curvature abnormalities among students of primary and secondary schools in Guizhou Province, so as to provide a basis for formulating school health policies and scientific prevention and control measures for abnormal spinal curvature in students. Methods: Using a stratified random sampling method, 471 schools (2 811 classrooms) were selected annually across the province from 2022 to 2024 in order to monitor and evaluate classroom environment according to national standards. Spinal examinations were conducted for 196 606 (2022), 194 876 (2023), and 195 048 (2024) students, and χ 2 test was used to analyze the correlation of desk and chair compliance with student spinal curvature abnormalities. Results: The qualified rates of classroom blackboard illuminance uniformity ( 85.41 %) and the blackboard size (82.24%) were the highest in primary and secondary schools in Guizhou Province, while the average blackboard surface illumination qualified rate (20.10%) was the lowest. The average desk illumination (50.11%-58.63%), desk illumination uniformity (61.92%-72.27%) and qualified noise rate (50.04%-51.94%) increased significantly in 3 years; in addition, the compliance rate of desks and chairs decreased from 25.00% to 13.52%, and the differences were all statistically significant ( χ 2=42.48, 80.93, 46.09, 129.72, all P <0.05). Middle school classrooms outperformed primary schools in terms of per capita area, average blackboard illuminance, blackboard illuminance uniformity, average desktop illuminance, desktop illuminance uniformity, and noise compliance ( χ 2=311.55, 12.41, 20.64, 40.76, 10.25, 52.47), but had lower compliance for blackboard size and reflectance ( χ 2=537.29, 7.59) (all P <0.01). Urban schools had higher compliance than suburban schools for per capita area, average blackboard illuminance, average desktop illuminance, and desktop illuminance uniformity ( χ 2=73.71, 17.68, 29.30 , 36.03), but lower compliance for desk-chair suitability, blackboard size, and blackboard reflectance ( χ 2=4.72, 26.02, 5.43 ) (all P <0.05). The spinal curvature abnormality detection rate was 0.83%. A significant association was found between abnormality detection and desk-chair non compliance ( χ 2=223.85, P <0.01). Conclusions Classroom environment hygiene in Guizhou schools is suboptimal. Strengthening school environmental hygiene infrastructure and greater attention to its impact on student s health are essential.

Objective: To understand the epidemiological characteristics of abnormalities spinal curvature among children and adolescents in Karst landform in Guizhou Province, so as to provide a scientific basis for targeted comprehensive intervention. Methods: From September to December 2023, 194 875 children and adolescents aged 6-18 were selected from all countieldistricts of 9 cities (prefectures) in Guizhou Province by stratified random cluster sampling method according to the proportion of Karst landform area in Guizhou Province for carrying out spinal curvature abnormality screening, and a questionnaire survey was conducted on 139 449 students in the fourth grade and above of primary school. Binary Logistic regression was used for multivariate analysis. Results: The detection rate of abnormal spinal curvature among children and adolescents aged 6-18 in Guizhou Province was 1.13%. The detection rates of abnormal spinal curvature in areas with a high proportion of Karst landform, areas with a low proportion of Karst landform, intercalation areas, and nonKarst landform areas were 1.10%, 2.00%, 0.90%, and 0.60%, respectively. Among them, the detection rates of abnormal spinal curvature in female students (2.40%, 1.60%, 0.90%) in areas with a low proportion of Karst landform, intercalation areas and nonKarst landform were higher than those in male students (1.60%, 0.10%, 0.30) (χ2=12.66, 112.69, 30.22, all P<0.05). The detection rates of abnormal spinal curvature among senior high school students (2.00%, 4.30%, 1.40%, 1.30%) in different Karst landform distributions were successively higher than those among junior high school students (1.40%, 3.20%, 1.00%, 0.60%) and primary school students (0.70%, 0.80%, 0.60%, 0.30%) (χ2=306.11, 175.80, 14.24, 39.57, all P<0.05). The results of multivariate Logistic regression analysis showed that the detection rates of abnormal spinal curvature in both highproportion and lowproportion Karst landform areas were higher than those in nonKarst landform areas [OR(95%CI)=1.84(1.05-2.25), 1.60(1.23-2.09), both P<0.05]. Conclusion The detection rate of abnormal spinal curvature in children and adolescents aged 6-18 in Guizhou Province is related to the distribution of Karst landform, so it is necessary to strengthen screening and appropriate comprehensive interventions.

Objective To mine and analyze the routine blood test data of children with allergic rhinitis (AR), identify routine blood parameters related to childhood allergic rhinitis, establish an effective diagnostic model, and evaluate the performance of the model. Methods This study was a retrospective study of clinical cases. The experimental group comprised a total of 1110 children diagnosed with AR at the First Affiliated Hospital of Air Force Medical University during the period from December 12, 2020 to December 12, 2021, while the control group included 1109 children without a history of allergic rhinitis or other allergic diseases who underwent routine physical examinations during the same period. Information such as age, sex and routine blood test results was collected for all subjects. The levels of routine blood test indicators were compared between AR children and healthy children using comprehensive intelligent baseline analysis, with indicators of P≥0.05 excluded; variables were screened by Lasso regression. Binary Logistic regression was used to further evaluate the influence of multiple routine blood indexes on the results. Five kinds of machine model algorithms were used, namely extreme value gradient lift (XGBoost), logistic regression (LR), gradient lift decision tree (LGBMC), Random forest (RF) and adaptive lift algorithm (AdaBoost), to establish the diagnostic models. The receiver operating characteristic (ROC) curve was used to screen the optimal model. The best LightGBM algorithm was used to build an online patient risk assessment tool for clinical application. Results Statistically significant differences were observed between the AR group and the control group in the following routine blood test indicators: mean cellular hemoglobin concentration (MCHC), hemoglobin (HGB), absolute value of basophils (BASO), absolute value of eosinophils (EOS), large platelet ratio (P-LCR), mean platelet volume (MPV), platelet distribution width (PDW), platelet count (PLT), absolute values of leukocyte neutrophil (W-LCC), leukocyte monocyte (W-MCC), leukocyte lymphocyte (W-SCC), and age. Lasso regression identified these variables as important predictors, and binary Logistic regression further analyzed the significant influence of these variables on the results. The optimal machine learning algorithm LightGBM was used to establish a multi-index joint detection model. The model showed robust prediction performance in the training set, with AUC values of 0.8512 and 0.8103 in the internal validation set. Conclusion The identified routine blood parameters can be used as potential biomarkers for early diagnosis and risk assessment of AR, which can improve the accuracy and efficiency of diagnosis. The established model provides scientific basis for more accurate diagnostic tools and personalized prevention strategies. Future studies should prospectively validate these findings and explore their applicability in other related diseases.
Humans ; Male ; Female ; Rhinitis, Allergic/blood* ; Child ; Biomarkers/blood* ; Retrospective Studies ; Early Diagnosis ; Child, Preschool ; ROC Curve ; Logistic Models ; Hematologic Tests ; Algorithms ; Adolescent ; Machine Learning

Nipah virus (NiV) and related viruses form a distinct henipavirus genus within the Paramyxoviridae family. NiV continues to spillover into the humans causing deadly outbreaks with increasing human-bat interaction. NiV encodes the large protein (L) and phosphoprotein (P) to form the viral RNA polymerase machinery. Their sequences show limited homologies to those of non-henipavirus paramyxoviruses. We report two cryo-electron microscopy (cryo-EM) structures of the Nipah virus (NiV) polymerase L-P complex, expressed and purified in either its full-length or truncated form. The structures resolve the RNA-dependent RNA polymerase (RdRp) and polyribonucleotidyl transferase (PRNTase) domains of the L protein, as well as a tetrameric P protein bundle bound to the L-RdRp domain. L-protein C-terminal regions are unresolved, indicating flexibility. Two PRNTase domain zinc-binding sites, conserved in most Mononegavirales, are confirmed essential for NiV polymerase activity. The structures further reveal anchoring of the P protein bundle and P protein X domain (XD) linkers on L, via an interaction pattern distinct among Paramyxoviridae. These interactions facilitate binding of a P protein XD linker in the nucleotide entry channel and distinct positioning of other XD linkers. We show that the disruption of the L-P interactions reduces NiV polymerase activity. The reported structures should facilitate rational antiviral-drug discovery and provide a guide for the functional study of NiV polymerase.
Nipah Virus/chemistry* ; Cryoelectron Microscopy ; Viral Proteins/genetics* ; RNA-Dependent RNA Polymerase/genetics* ; Phosphoproteins/genetics* ; Humans ; Models, Molecular ; Protein Binding

N⁶-methyladenosine (m⁶A) modification plays a critical role in cell cycle regulation, while the mechanism of m⁶A in regulating mitosis remains underexplored. Here, we found that the total m⁶A modification level in cells increased during mitosis by the liquid chromatography-mass spectrometry/mass spectrometry and m⁶A dot blot assays. Silencing methyltransferase-like 3 (METTL3) or METTL14 results in delayed mitosis, abnormal spindle assembly, and chromosome segregation defects by the immunofluorescence. By analyzing transcriptome-wide m⁶A targets in HeLa cells, we identified polo-like kinase 1 (PLK1) as a key gene modified by m⁶A in regulating mitosis. Specifically, through immunoblotting and RNA pulldown, m⁶A modification inhibits PLK1 translation via YTH N⁶-methyladenosine RNA binding protein 1, thus mediating cell cycle homeostasis. Demethylation of PLK1 mRNA leads to significant mitotic abnormalities. These findings highlight the critical role of m⁶A in regulating mitosis and the potential of m⁶A as a therapeutic target in proliferative diseases such as cancer.
Humans ; Polo-Like Kinase 1 ; Cell Cycle Proteins/metabolism* ; Proto-Oncogene Proteins/metabolism* ; Protein Serine-Threonine Kinases/metabolism* ; Mitosis/physiology* ; HeLa Cells ; Adenosine/genetics* ; Methyltransferases/metabolism* ; RNA, Messenger/metabolism* ; RNA-Binding Proteins/metabolism*

Objective To investigate the therapeutic efficacy of artesunate(AS)on polycystic ovary syndrome(PCOS)in mice and explore the potential mechanism primarily.Methods Twenty-five female C57BL/6J mice were randomly divided into Control group,model group(PCOS group),low-and high-dose AS groups(AS15 and AS30 groups)and metformin group(Met group).In addition to the Control group,the mouse model of PCOS was established by subcutaneous injection of dehydroepiandrosterone(DHEA,60 mg/kg)following by a high-fat diet for 21 d.After modeling,AS of 15 and 30 mg/kg was intraperitoneally injected into the mice of the AS 15 and AS30 groups,respectively,and 200 mg/kg Met was given to those of the Met group by gavage,once per day,for 6 weeks.ELISA was used to detect serum testosterone(T),fasting insulin(FINS),luteinizing hormone(LH)and follicle-stimulating hormone(FSH),and the LH/FSH ratio was calculated.The levels of fasting blood glucose(FBG),triglyceride(TG)and total cholesterol(TC)were detected by automatic biochemical analyzer,and the homeostasis model assessment of insulin resistance(HOMA-IR)was calculated.The estrous cycle was observed,and HE staining was performed for pathological changes in the ovary and uterus.Immunofluorescence assay was employed to measure the expression of p-eIF2α,ATF4 and CHOP in the ovarian tissue.After steroidogenic human granulosa-like tumor cell line KGN were exposed to 100 μmol/L DHEA to simulate the hyperandrogen environment of PCOS,and then treated with 5 and 10 μg/mL AS for 24 h,the protein levels of endoplasmic reticulum stress signaling pathway was detected by Western blotting.Results Compared with the Control group,the PCOS mice had disturbed estrous cycle,polycystic changes in the ovaries,and significantly increased serum T level and LH/FSH ratio(P＜0.05),and obviously elevated HOMA-IR,TC and TG levels in terms of metabolism(P＜0.01).The expression levels of p-eIF2α,ATF4 and CHOP were notably up-regulated in the ovarian granulosa cells of PCOS mice and KGN cells after DHEA exposure(P＜0.05).Additionally,AS treatment attenuated the pathological changes of ovary and uterine expression,decreased the serum T level and the LH/FSH ratio(P＜0.05),and reduced HOMA-IR,TC and TG levels(P＜0.05)when compared with the PCOS mice.Moreover,the expression levels of p-eIF2α,ATF4 and CHOP were significantly down-regulated after AS treatment in both ovarian granulosa cells of PCOS mice and KGN cells(P＜0.05).Conclusion AS significantly improves glycolipid metabolic disorder and reproductive dysfunction in PCOS mice,which may be associated with its suppressing endoplasmic reticulum stress by inhibiting the PERK/eIF2α/ATF4/CHOP pathway.

Objective To investigate the role of transcription factor EB(TFEB)in endotoxin-tolerant macrophages.Methods The RAW264.7 cells were divided into blank group(DMEM medium),LPS 5 group(5 ng/mL LPS treatment for 4 h),LPS 100 group(100 ng/mL LPS treatment for 4 h),and tolerance group(5 ng/mL LPS for 12 h followed by 100 ng/mL LPS for 4 h).The releases of inflammatory factors TNF-α and IL-6 were measured using ELISA.Western blotting and immunofluorescence assay were used to evaluate the distribution of autophagy-related proteins LC3 and P62,as well as TFEB in the cytoplasm and nucleus.Lentiviral overexpression of TFEB or siRNA-mediated knockdown of TFEB were performed to observe the changes in autophagy levels and bacterial clearance ability in the tolerant cells.Results The cells in the tolerance group had significantly lower contents of TNF-α and IL-6,as well as reduced bacterial clearance ability(P<0.01),down-regulated LC3 expression while up-regulated P62 level,and decreased expression of TFEB in both the cytoplasm and nucleus(P<0.01)when compared with the cells of the LPS 100 group.Overexpression of TFEB significantly increased LC3 level,reduced P62 level,and enhanced bacterial clearance ability in the endotoxin-tolerant cells(P<0.01).In contrast,siRNA-mediated knockdown of TFEB had no significant impacts on LC3 and P62 expression levels or bacterial clearance ability.Conclusion Overexpression of TFEB can restore the autophagy of endotoxin-tolerant cells and enhance their bacterial clearance capacity,thereby alleviating the immunosuppressive state of sepsis.These findings suggest that TFEB holds promise as a potential therapeutic target for the prevention and treatment of sepsis.

Objective To investigate the role and underlying mechanism of activating transcription factor 3(ATF3)in suppressing lipopolysaccharide(LPS)-induced autophagy and inflammatory responses in macrophages.Methods Firstly,the gene expression omnibus(GEO)database was used to analyze ATF3 expression in peripheral blood mononuclear cells(PBMCs)from sepsis patients,and gene set enrichment analysis(GSEA)was performed to identify enriched signaling pathways.Secondly,RAW264.7 macrophages were divided into a blank control group and an LPS-stimulated group(100 ng/mL LPS).Western blotting and immunofluorescence assay were used to detect ATF3 protein expression and observe its subcellular localization,respectively.Lentiviral transduction was used to generate ATF3 knockdown and overexpression cell lines to evaluate their effects on cytokine release and bacterial clearance.Cleavage Under Targets and Tagmentation(CUT&Tag)sequencing was employed to identify downstream target genes transcriptionally regulated by ATF3.Furthermore,the impact of ATF3 knockdown or overexpression on autophagy-related gene 5(ATG5),autophagy-related gene 16-like 1(ATG16L1),and autophagy levels was evaluated.Results GEO analysis revealed that ATF3 expression was significantly elevated in PBMCs from sepsis patients(P<0.01),and GSEA showed significant enrichment of autophagy-related and inflammation-related pathways(P<0.01).In RAW264.7 cells,100 ng/mL LPS stimulation significantly increased ATF3 expression in the nucleus than the blank control group(P<0.01).ATF3 knockdown led to increased secretions of TNF-α and IL-6 and enhanced bacterial clearance of macrophages(P<0.01),whereas ATF3 overexpression significantly suppressed TNF-α and IL-6 releases,and remained bacterial clearance at a low level when compared with the conditions in the negative control(NC)group(P<0.01).CUT&Tag results demonstrated that ATF3 was enriched at the promoter regions of key autophagy genes Atg5 and Atg16l1.Compared with the NC group,ATF3 knockdown significantly up-regulated the protein levels of LC3-II/I,ATG5,and ATG16L1 while decreased p62 expression(P<0.01).Conversely,ATF3 overexpression inhibited the expression of LC3-II/I,ATG5,and ATG16L1(P<0.01),but had no significant effect on p62 level.Conclusion Sepsis induces elevated ATF3 expression in macrophages,and suppresses autophagic activity and down-regulates pro-inflammatory cytokines TNF-α and IL-6,which probably mediated by ATF3 regulating transcription of ATG5 and ATG16L1,suggesting ATF3 as a potential therapeutic target for autophagy-inflammation imbalance.

BACKGROUND:Due to the small number of autologous bone sources and the risk of immune rejection and disease spread caused by the use of allogeneic bone,artificial bone materials have played an irreplaceable role in bone transplantation today.Along with functional customization,biocompatibility requirements,and the emergence of biodegradable materials,a variety of biomaterials and a variety of preparation methods have emerged. OBJECTIVE:To summarize the preparation methods of scaffolds used in bone tissue engineering,and the advantages and disadvantages,research status and progress of various preparation methods. METHODS:A computer search was conducted on CNKI,WanFang Data,PubMed,and ScienceDirect databases for literature related to bone tissue engineering scaffold from January 2008 to August 2023.Chinese and English search terms were"tissue engineering,bone scaffold,gas foaming,cryotropic gelation,additive manufacturing".After excluding irrelevant and repetitive studies,a total of 80 articles were retained for summary. RESULTS AND CONCLUSION:(1)Compared with the traditional preparation process of scaffolds,the emerging additive manufacturing and electrospinning technologies have shown great potential in the production of complex structures such as bone and cartilage for tissue engineering in recent years,demonstrating enormous potential.(2)In addition to the advantages of speed,precision and the range of materials used,additive manufacturing methods also provide the feasibility of manufacturing highly complex geometry and topologically optimized structures,achieving precise adjustment and high repeatability of the structure.(3)Electrospinning is one of the most adaptable and promising technologies for the production of a series of fiber mats.The nanofiber scaffolds produced by electrospinning are biomaterials with surprisingly similar microstructures to the cytoplasmic matrix.(4)At present,hydroxyapatite and tricalcium phosphate are the best in ceramic materials,and there are a variety of materials in polymer materials,with excellent biocompatibility.(5)Therefore,the selection of materials should be based on a better understanding of their properties,avoiding complexity,and producing more enhanced scaffolds.However,most of the literature reports so far are exploratory in terms of clinical applicability,and the specific diseases for which they are suitable for treatment remain to be tested.The future development of bone scaffolds is reflected in the following aspects:mechanical properties matching the missing bone,controllable degradation rate,strong ability to promote bone regeneration,and specific functions.

Objective:To investigate the expression of microRNA-34a (miR-34a)and tumor necrosis factor-α (TNF-α) in lung injury induced by lipopolysaccharides (LPS) in neonatal rats with acute respiratory distress syndrome (ARDS). The expression of mirna-34a and tumor necrosis factor-α (TNF-α) in lung injury induced by lipopolysaccharides (LPS) was determined by tumor necrosis factor.Methods:A total of 80 7-day-old newborn SD rats were randomly(random number) divided into two groups, 40 in each group, NARDS animal models were established by intraperitoneal injection of 4 mg/kg LPS solution for 3,6,12 and 24 hours respectively. Normal control group was established by intraperitoneal injection of 4 mL/kg isotonic Nacl solution. The wet/dry weight ratio of lung was measured to observe the changes of lung histopathology. The expression of miR-34a in lung tissues and cells was detected by quantitative real-time PCR (qrt-PCR).Results:the wet/dry weight of lung in the experimental group was significantly higher than that in the control group. Hematoxylin-eosin (HE) staining showed diffuse changes in lung tissue, thickening of alveolar septum, partial alveolar fusion and decreased number of alveoli in experimental group. The expression levels of miR-34a and TNF-α in the experimental group were higher than those in the control group at each time point.Conclusions:It is speculated that miR-34a can promote the release of more inflammatory factors by positively regulating the expression of TNF-α, thus affecting the occurrence and development of lung injury. The expression of miR-34a positively correlated with TNF-α. miR-34a may affect the regulation mechanism of lung injury by mediating the expression of TNF-α through some signal pathway or inflammatory reaction, it provides a new therapeutic target for the treatment of neonatal acute respiratory distress syndrome (NARDS).