ObjectiveTo investigate the effects of Astragali Radix polysaccharides （APS） on the polarization of BV2 microglial cells in an oxygen-glucose deprivation/reoxygenation （OGD/R） model through regulation of the Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB） signaling pathway. MethodsThe OGD/R injury model of BV2 microglia was established and divided into blank group， OGD/R group and APS group （0.4 g·L^-1 APS）. Neuroinflammatory injury was induced by lipopolysaccharide （LPS） and treated with APS. The cells were divided into blank group， LPS group （1 mg·L^-1 LPS） and APS group （0.4 g·L^-1 APS+1 mg·L^-1 LPS）. Cell viability was detected using the cell counting kit-8 （CCK-8） assay. Cell morphology was observed under an inverted microscope. Nitric oxide （NO） content in the cell supernatant was determined by the Griess assay. The secretion levels of tumor necrosis factor-α （TNF-α）， interleukin （IL）-6， IL-10， and IL-4 were measured by enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence （IF） was used to detect the double-positive rates of ionized calcium-binding adapter molecule-1/inducible nitric oxide synthase （Iba-1⁺/iNOS⁺） and ionized calcium-binding adapter molecule-1/arginase 1 （Iba-1⁺/Arg1⁺）， as well as the nuclear translocation rate of nuclear factor-κB p65 （NF-κB p65）. Protein expression levels of Iba-1， iNOS， Arg1， TLR4， and NF-κB p65 were detected by Western blot. ResultsIn the OGD/R injury model， compared with the blank control group， BV2 microglial cells in the OGD/R group were activated and exhibited amoeboid morphological changes. The secretion levels of NO， TNF-α， and IL-6 were significantly increased （P<0.01）. The double-positive expression rate of Iba-1⁺/iNOS⁺ and the protein expression of Iba-1 and iNOS were significantly increased （P<0.01）. The nuclear translocation rate of NF-κB p65 and the protein expression levels of TLR4 and NF-κB p65 were significantly increased （P<0.01）. The levels of IL-10 and IL-4 were significantly decreased （P<0.01）， and the double-positive expression rate of Iba-1⁺/Arg1⁺ and Arg1 protein expression were significantly decreased （P<0.01）. Compared with the OGD/R group， the APS group （0.4 g·L^-1） showed reduced cell activation， significantly decreased secretion levels of NO， TNF-α， and IL-6 （P<0.01）， significantly decreased double-positive expression rate of Iba-1⁺/iNOS⁺ and relative protein expression of Iba-1 and iNOS （P<0.01）， significantly decreased nuclear translocation rate of NF-κB p65 and protein expression levels of TLR4 and NF-κB p65 （P<0.01）， significantly increased levels of IL-10 and IL-4 （P<0.01）， and significantly increased double-positive expression rate of Iba-1⁺/Arg1⁺ and Arg1 protein expression （P<0.01）. In the LPS-induced neuroinflammation model， compared with the blank control group， the LPS group showed increased cell activation， significantly increased levels of NO， TNF-α， and IL-6， significantly increased Iba-1⁺/iNOS⁺ double-positive expression rate， NF-κB p65 nuclear translocation rate， and protein expression levels of Iba-1， iNOS， TLR4， and NF-κB p65 （P<0.01）， while IL-10 and IL-4 levels， Iba-1⁺/Arg1⁺ double-positive expression rate， and Arg1 protein expression were significantly decreased （P<0.01）. Compared with the LPS group， the APS group showed reduced cell activation， significantly decreased levels of NO， TNF-α， and IL-6， Iba-1⁺/iNOS⁺ double-positive expression rate， NF-κB p65 nuclear translocation rate， and protein expression levels of Iba-1， iNOS， TLR4， and NF-κB p65 （P<0.01）， while IL-10 and IL-4 levels， Iba-1⁺/Arg1⁺ double-positive expression rate， and Arg1 protein expression were significantly increased （P<0.01）. ConclusionAPS may reduce microglial activation and promote their polarization toward the M2 phenotype by inhibiting activation of the TLR4/NF-κB signaling pathway， thereby alleviating the neuroinflammatory response induced by OGD/R.

Objective To establish a fluorescent recombinase-aided amplification (RAA) assay for detection of Strongyloides stercoralis nucleic acid and to preliminarily evaluate its performance. Methods Six sets of specific primers targeting S. stercoralis 18S ribosomal RNA (18S rRNA) gene and one fluorescent probe were designed and synthesized. The optimal primer-probe set was determined through systematic screening and optimization to establish the fluorescent RAA assay. The assay was evaluated using S. stercoralis genomic DNA at concentrations of 100, 10, and 1 pg/μL, and 100, 10, and 1 fg/μL, as well as recombinant pUC57 plasmids containing the target gene fragments at 1 × 10⁵, 1 × 10⁴, 1 × 10³, 1 × 10², 1 × 10¹, 1 × 10⁰ copies/reaction, to determine the analytical sensitivity. Genomic DNA from Ascaris lumbricoides, Ancylostoma duodenale, Enterobius vermicularis, Angiostrongylus cantonensis, Trichinella spiralis, Clonorchis sinensis, Schistosoma japonicum, and Taenia saginata was used to assess assay specificity. A total of 25 stool samples from patients suspected of S. stercoralis infection were tested by the modified Baermann funnel technique, PCR, and the established fluorescent RAA assay. The sensitivity, specificity, concordance rate and their 95% confidence intervals (CI) of these three techniques were estimated, and agreement between methods was evaluated using the Kappa coefficient. Results Exo-4 was identified as the optimal primer set screened from the six primer sets, and the best amplification performance was achieved when the final concentrations of the forward and reverse primers were 0.44 μmol/L and a probe concentration was 0.20 μmol/L. The limit of detection of the fluorescent RAA assay was 100 fg/μL for genomic DNA of S. stercoralis and 1 × 10⁰ copies/reaction for recombinant plasmids. Specific fluorescence signals were detected within 5 min, with no cross-reactivity observed with A. lumbricoides, A. duodenale, E. vermicularis, A. cantonensis, T. spiralis, C. sinensis, S. japonicum, or T. saginata. Among the 25 clinical stool samples from patients suspected of S. stercoralis infections, the modified Baermann funnel technique and fluorescent RAA assay detected 19 positives and 6 negatives, whereas PCR detected 18 positives and 7 negatives. The fluorescent RAA assay showed a sensitivity of 100.00% [95% CI: (82.35%, 100.00%)], specificity of 100.00% [95% CI: (54.07%, 100.00%)], concordance rate of 100.00% [95% CI: (86.28%, 100.00%)], and a Kappa coefficient of 1.00 [95% CI: (1.00, 1.00)] (P < 0.001) relative to the modified Baermann funnel technique, and a sensitivity of 100.00% [95% CI: (81.47%, 100.00%)], specificity of 85.71% [95% CI: (42.13%, 99.64%)], concordance rate of 96.00% [95% CI: (79.65%, 99.90%)], and a Kappa coefficient of 0.90 [95% CI: (0.70, 1.00)] (P < 0.001). Positive amplification products emitted green fluorescence under a portable blue-light device, enabling visual interpretation of results. Conclusions The fluorescent RAA assay established in this study is rapid, highly sensitive, and highly specific. It enables detection of S. stercoralis nucleic acid under isothermal conditions and allows visual interpretation of results, providing a novel tool for rapid clinical diagnosis and field screening of S. stercoralis infections.

Objective: To evaluate the efficacy of a short video based health education model in enhancing e-cigarette awareness among junior high school students, so as to provide empirical evidence for optimizing school based health education in the digital era. Methods: In December 2024, a cluster sampling method was applied to recruit 465 eighth grade students from a middle school in Kunming for a survey. The intervention was implemented through playing short videos produced by the research team in the classroom, and a self designed adolescent e-cigarette questionnaire was used to conduct assessments before and after intervention, along with qualitative interviews with 60 teachers and students by purposive sampling. Pre and post intervention outcomes were compared by using paired t-test and Chi-square test. Qualitative data underwent a three stage coding process of "open, axial, and selective coding" to construct an explanatory framework. Results: Quantitative analysis showed significant post intervention increases in both baseline awareness scores（92.1±6.6）and deep awareness scores (70.0±17.6) compared with pre intervention( 78.2± 14.0，46.1±20.5)( t =23.54，26.72，both P <0.05), both meeting the criteria for large effect sizes (Cohen s d =1.09, 1.24). The improvement in deep awareness was particularly pronounced, specifically regarding addictive ingredients, illicit additives(tiletamine,metomidate,etomidate), and misconceptions about smoking cessation efficacy ( χ 2=150.54,113.27,117.40,112.92, 69.93 , all P <0.01). Qualitative insights revealed that narrative transportation and visual enhancement were the core mechanisms of efficacy: 87.3% of students preferred contextualized narratives, 85.5% of students decoded cognitive bottlenecks more effectively because of visual enhancement, and 72.7% relied on short videos for micro learning. Conclusion Short video based health education can effectively enhance e-cigarette awareness among junior high school students, offering a pragmatically viable digital pathway for school based health education.

Objective:To evaluate alterations in dopaminergic neurons, cortical metabolism, and functional connectivity networks in patients with early-onset Parkinson′s disease (EOPD) using multimodal neuroimaging.Methods:In this prospective cross-sectional study, 26 patients with EOPD and 16 healthy controls (HC group) were recruited from the PLA General Hospital between April and November 2023. All participants underwent integrated 11C-β-CFT PET/MR, 18F-FDG PET/CT brain imaging and resting-state functional MRI. Clinical assessments were conducted using the Unified Parkinson′s Disease Rating Scale and Hoehn-Yahr staging. Cognitive status was evaluated using the Mini-Mental State Examination and Montreal Cognitive Assessment. Standardized uptake value ratios for both 11C-β-CFT and 18F-FDG PET images were calculated using cerebellar gray matter as the reference region. Voxel-wise two-sample t-tests were performed to identify regions with significant group differences in tracer uptake. Seed regions showing altered 11C-β-CFT or 18F-FDG uptake were used to compute seed-based functional connectivity (FC) with all other brain voxels, and group differences in FC were assessed. Correlations between imaging metrics and clinical scales were evaluated using Pearson or Spearman analyses as appropriate. Results:Compared with HC group, EOPD group showed significantly reduced 11C-β-CFT uptake in the bilateral putamen, globus pallidus, and left temporal pole ( P<0.05), and decreased 18F-FDG uptake in the right superior frontal gyrus and anterior cingulate cortex ( P<0.05). Relative to HC group, EOPD group exhibited markedly lower FC between the right putamen and the left gyrus rectus as well as the right parahippocampal gyrus; the right superior frontal gyrus and the left gyrus rectus; the anterior cingulate cortex and the olfactory area of the frontal lobe, the left gyrus rectus, and the right superior parietal gyrus; the left temporal pole and the left orbitofrontal cortex as well as the left olfactory area ( P<0.05). Correlation analyses revealed no statistically significant associations between altered FC values and clinical scale scores in the EOPD group. Conclusions:Patients with EOPD demonstrate impaired nigrostriatal dopaminergic function, regional cortical hypometabolism, and aberrant functional connectivity across multiple brain networks.

Purpose To explore the expression level of let-7b-5p in glioma and its effects and potential mecha-nisms on U251 cell growth.Methods The expression of let-7b-5p in glioma was detected using qRT-PCR.Data from the CGGA database were analyzed to examine the relationship between the let-7b-5p expression levels,WHO grade and overall survival rates of glioma patients.Transient transfection was used to downregulate the expression of let-7b-5p and IGF1R in U251 cells.The role and potential mechanism of let-7b-5p in the U251 cell were evaluated using qRT-PCR,CCK8 assays,clone formation assays,Western blotting,and double luciferase reporter assays.Results The expres-sion of let-7b-5p in glioma cells(A172:3.64±0.64,V251:4.56±0.52,U87-MG:3.31±0.50)and tissues(2.18±0.22)was significantly higher than that in astrocytes(HMC3:1.00±0.21,P＜0.05 or P＜0.01)and nor-mal brain tissues(1.01±0.19,P＜0.05).Let-7b-5p expression was negatively correlated with WHO grades but pos-itively correlated with survival rates in primary and recurrent glioma patients(P＜0.000 1 and P=0.028,respective-ly).Knockdown of let-7b-5p in U251 cells significantly promoted the growth of glioma cells(CCK8:knockdown group 126.00±12.09 vs miR-NC group 90.93±5.13,P＜0.05)and activated PI3K/AKT signal pathway.Suppressing IGF1R expression in U251 cells reversed the effects of let-7b-5p knockdown on glioma cell growth[CCK8:let-7b-5p knockdown+IGF1R knockdown group(92.08±6.14)vs let-7b-5p knockdown+sh-NC group(116.67.08±8.50)]and PI3K/AKT signal pathway activation.Conclusion Let-7b-5p functions as a tumor suppressor gene in glioma.It may regulate glioma cell growth by targeting IGF1R and modulating PI3K/AKT signal pathway.

Cardiac arrest leads to abrupt cessation of cardiac ejection and effective systemic circulation,placing patients in a life-threatening state.Despite advancements in cardiopulmonary resuscitation(CPR)techniques significantly improving the rate of return of spontaneous circulation,overall hospital discharge rates remain low,primarily due to severe hypoxic-ischemic encephalopathy following CPR,which is the leading cause of mortality or prolonged coma.Accurately predicting neurological prognosis post-CPR is critical for optimizing therapeutic strategies and improving long-term outcomes.This review systematically explores diverse methods for predicting neurological prognosis after CPR,encompassing traditional neurological examinations,cerebral performance category scales,hemodynamic monitoring,and advanced approaches such as neuroimaging,serum biomarkers,neuroelectrophysiological assessments,inflammatory indicators,brain-computer interface technology,and multimodal detection.These methods collectively reflect the severity of brain injury and potential for functional recovery.Neurological examinations and cerebral performance category scales provide rapid initial assessments of functional status;neuroimaging visually delineates the extent of structural brain damage;serum biomarkers and inflammatory indicators evaluate metabolic and immune responses to injury;neuroelectrophysiological monitoring captures real-time neuronal activity.Furthermore,emerging brain computer interface technology and multimodal approaches integrate multiple modalities,offering novel possibilities for precise assessment and dynamic monitoring.By synthesizing reliable prognostic tools,this review facilitates early and accurate evaluation of brain injury severity and recovery potential,guiding timely clinical interventions and providing evidence-based references for post-CPR.

This study aims to explore protective effects of Lycium barbarum polysaccharides(LBP)on intestinal damage caused by lipopolysaccharide(LPS)-induced inflammatory bowel disease(IBD)in chicks.Network pharmacology was initially employed to determine the target proteins of wolfberry in the prevention and treatment of IBD.Following this,protein-protein interaction analy-sis,GO and KEGG pathway enrichment analysis,and molecular docking studies were conducted.Subsequently,an animal study was conducted:a total of 100 one-day-old male Hy-line brown lay-ing hens were randomly divided into five groups:a blank control group(CON),an LPS treatment group(LPS),a low-dose LBP group(LPS+LBP 0.25 g/L,L-LBP),a medium-dose LBP group(LPS+LBP 0.5 g/L,M-LBP),and a high-dose LBP group(LPS+LBP 1 g/L,H-LBP).Upon reac-hing 21 days old,duodenal,jejunal,ileal,and cecal tissues were collected to determine SOD and GSH-Px levels.Furthermore,the mRNA expression levels of TNF-α,AKT1,IL-6,IL-1β and TP53 in the intestinal tissues were measured using quantitative real-time PCR.The results demonstrated that network pharmacology identified 45 active ingredients in wolfberry that target 116 key protein sites,including TNF,AKT1 and IL6.The primary objectives focus on signaling pathways including AGE-RAGE,IL-17,TNF,HIF-1,and NF-κB.Molecular docking showed excellent ligand-receptor docking scores,with stable binding facilitated by hydrogen bonds and hydrophobic interactions.Compared to the LPS group,the 0.5 g/L LBP exhibited notably higher levels of SOD and T-AOC.In comparison with the LPS group,the medium and high-dose LBP experimental groups showed notably decreased the mRNA expressions of TNF-α,AKT1,IL-6,and IL-1β,while TP53 mRNA expression was significantly upregulated(P＜0.01).In summary,wolfberry exerts preventive and therapeutic effects on IBD through a multi-component,multi-target,and multi-pathway mecha-nism.

This study aims to explore protective effects of Lycium barbarum polysaccharides(LBP)on intestinal damage caused by lipopolysaccharide(LPS)-induced inflammatory bowel disease(IBD)in chicks.Network pharmacology was initially employed to determine the target proteins of wolfberry in the prevention and treatment of IBD.Following this,protein-protein interaction analy-sis,GO and KEGG pathway enrichment analysis,and molecular docking studies were conducted.Subsequently,an animal study was conducted:a total of 100 one-day-old male Hy-line brown lay-ing hens were randomly divided into five groups:a blank control group(CON),an LPS treatment group(LPS),a low-dose LBP group(LPS+LBP 0.25 g/L,L-LBP),a medium-dose LBP group(LPS+LBP 0.5 g/L,M-LBP),and a high-dose LBP group(LPS+LBP 1 g/L,H-LBP).Upon reac-hing 21 days old,duodenal,jejunal,ileal,and cecal tissues were collected to determine SOD and GSH-Px levels.Furthermore,the mRNA expression levels of TNF-α,AKT1,IL-6,IL-1β and TP53 in the intestinal tissues were measured using quantitative real-time PCR.The results demonstrated that network pharmacology identified 45 active ingredients in wolfberry that target 116 key protein sites,including TNF,AKT1 and IL6.The primary objectives focus on signaling pathways including AGE-RAGE,IL-17,TNF,HIF-1,and NF-κB.Molecular docking showed excellent ligand-receptor docking scores,with stable binding facilitated by hydrogen bonds and hydrophobic interactions.Compared to the LPS group,the 0.5 g/L LBP exhibited notably higher levels of SOD and T-AOC.In comparison with the LPS group,the medium and high-dose LBP experimental groups showed notably decreased the mRNA expressions of TNF-α,AKT1,IL-6,and IL-1β,while TP53 mRNA expression was significantly upregulated(P＜0.01).In summary,wolfberry exerts preventive and therapeutic effects on IBD through a multi-component,multi-target,and multi-pathway mecha-nism.

Objective:To investigate the current status of fertility concerns among early-stage endometrial cancer (EC) and atypical endometrial hyperplasia (AEH) patients with fertility preservation, and analyze its influencing factors.Methods:Convenience sampling was used to select early-stage EC and AEH patients with fertility preservation at Peking University People's Hospital from January to December 2021 as study subjects. The study subjects were surveyed using the General Information Questionnaire, Chinese version of the Reproductive Concerns after Cancer Scale (RCAC), and Social Support Rating Scale (SSRS). Pearson correlation was employed to examine the relationship between fertility concerns and social support in early-stage EC and AEH patients with fertility preservation. Single-factor analysis and multiple linear regression analysis were used to examine the factors influencing fertility concerns in early-stage EC and AEH patients with fertility preservation.Results:A total of 170 questionnaires were distributed, and 167 valid questionnaires were collected, with a valid response rate of 98.24% (167/170). The RCAC and SSRS scores of 167 early-stage EC and AEH patients with fertility preservation were (56.58±10.58) and (34.22±8.21), respectively. Educational level, disease type and staging, marital status, and social support were statistically significant factors influencing fertility concerns among early-stage EC and AEH patients undergoing fertility preservation ( P<0.05), explaining 32.80% of the total variance. Conclusions:Early-stage EC and AEH patients with fertility preservation exhibit high levels of fertility concerns. Clinical practice should develop individualized psychological intervention programs for patients with high education level, unmarried status, high pathological grading, and lack of social support to improve their physical and mental health.

Objective:To prepare domestic 64Cu-1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid- D-Phe1-Tyr3-Thr8-octreotide (DOTATATE), and to verify its distribution and preliminary diagnostic value in neuroendocrine tumors (NETs). Methods:DOTATATE was labeled with the domestic 64Cu to obtain 64Cu-DOTATATE. The lipophilicity, in vitro stability, and pharmacokinetics were studied. Biodistribution experiments and microPET imaging were performed on NCI-H727 (somatostatin receptor (SSTR)2 positive expression) tumor-bearing nude mice. The preliminary clinical applications were conducted on 10 NETs patients (5 males, 5 females; age (58.5±13.0) years) from Chinese PLA General Hospital between May 2023 and April 2024. Data were analyzed by using independent-sample t test. Results:64Cu-DOTATATE was successfully prepared with the radiochemical purity greater than 98%, log P of -2.609±0.051 and good stability. Pharmacokinetic experiments in BALB/c mice suggested rapid blood clearance of the drug (elimination half-time of 22.78min). Biodistribution results in tumor-bearing mice showed that 64Cu-DOTATATE was mainly metabolized through the liver and kidneys, with significant tumor uptake at 1h ((2.519±0.273) percentage activity of injection dose per gram of tissue (%ID/g)) and sustained high uptake at 24h ((4.331±0.549)%ID/g). MicroPET imaging of tumor-bearing mice showed a slight increase in uptake and good retention at 24h, with a significant statistical difference compared to the blocked group ((2.197±0.250) vs (0.985±0.064) % ID/g; t=6.40, P=0.008). The tumor/liver ratios were 0.075±0.007, 0.083±0.011, 0.118±0.005, 0.263±0.031 at 1, 2, 6 and 24h, respectively. Preliminary clinical application indicated that 64Cu-DOTATATE exhibited good targeting in patients, and the liver radioactivity distribution was moderate (SUV max=10.62±3.46), providing good image quality. Conclusion:Domestic 64Cu-DOTATATE PET/CT imaging is a promising imaging evaluation method in NETs with the value for further clinical research.