Objective: To assess the effect of 12-year-old children s pit and fissure sealants on the health of first permanent molars, so as to provide evidence for optimizing caries prevention strategies among children. Methods: In March 2025, a cluster random sampling method was used to conduct oral examinations on 965 students aged 12 from Chengdu s 2021 Comprehensive Intervention Program for Pediatric Oral Diseases. Data from the Comprehensive Intervention System for Children s Oral Diseases were referenced. Participants were divided into a sealed group ( n =755) and an unsealed group ( n =210) based on whether they had received sealants on their first permanent molars. Chi square test or analysis of variance were used to compare indicators such as caries incidence, new caries detection rate, and new caries mean (DMFT increment) between the two groups Results: The sealed group showed significantly lower caries incidence, new caries detection rate, and new caries mean (33.38%, 17.65%, 0.59±1.00) compared to the unsealed group (43.81%, 24.70%, 0.87±1.22)( χ 2/F =7.79, 18.26, 9.55, all P <0.05). However, no significant difference was found in the filled teeth ratio between the two groups (20.38% , 20.16%; χ 2=0.01, P =0.94). In girls, the sealed group exhibited significantly lower caries incidence, new caries detection rate, and new caries mean (36.78%, 20.99%, 0.69± 1.10 ) than the unsealed group (57.55%, 33.52%, 1.15±1.29) ( χ 2/F =14.42, 23.76, 10.92, all P <0.05), whereas no significant differences were observed between boys in the sealed (30.47%, 14.85%, 0.50±0.89) and unsealed groups (29.81%, 16.18%, 0.59± 1.08) ( χ 2/F =0.02, 0.41, 0.74, all P >0.05). Boys had significantly lower new caries detection rates and new caries means than girls in both groups ( χ 2/F =16.20, 6.94; 29.93, 11.84, all P <0.05). In urban areas, the sealed group had lower new caries detection rates and new caries means (19.37%, 0.68±1.04) than the unsealed group (24.66%, 0.90±1.20) ( χ 2/F =6.86, 3.94, both P <0.05). In suburban areas, all indicators for the sealed group (24.71%, 13.77%, 0.42±0.87) were significantly lower than those for the unsealed group (38.81%, 24.77%, 0.82±1.28) ( χ 2/F =5.28, 15.36, 6.00, all P <0.05). Indicators from specialized dental institutions (11.25%, 4.81%, 0.16±0.56) were significantly lower than those from county level or above general hospitals (33.33%, 19.11%, 0.38±1.00) and primary healthcare institutions (37.59%, 19.24%, 0.67±1.05) ( χ 2/F =20.99, 34.31, 21.08 , all P <0.01). Conclusions The 12-year-old children s pit and fissure sealants effectively reduce the caries incidence in first permanent molars, particularly showing significant effectiveness in girls and suburban children. Intervention strategies should be optimized according to gender.

Objective:To study the effects of fluoride and aluminum exposure alone and in combination on pyroptosis and the NOD-like receptor protein 3 (NLRP3)/cysteinyl aspartate specific proteinase-1 (Caspase-1)/gasdermin D (GSDMD) signaling pathway in NG108-15 cells.Methods:Using a factorial design method, NG108-15 cells cultured in vitro were divided into control group [0 mg/L sodium fluoride (NaF) + 0 mg/L aluminium trichloride (AlCl 3)], fluoride group (40 mg/L NaF + 0 mg/L AlCl 3), aluminum group (0 mg/L NaF + 160 mg/L AlCl 3), and fluoride + aluminum group (40 mg/L NaF + 160 mg/L AlCl 3) according to the concentrations of NaF and AlCl 3. After 24 hours of cultivation, the cells were collected for subsequent experiments. The fluorescence intensity of pyroptosis index GSDMD in each group was detected by immunofluorescence method. The mRNA expression levels of NLRP3, apoptosis associated speck-like protein (ASC), Caspase-1, and GSDMD in each group were detected by real-time fluorescence quantitative PCR (qRT-PCR). The protein expression levels of NLRP3, ASC, Caspase-1, GSDMD, gasdermin D N-terminus (GSDMD-N) and interleukin-1β (IL-1β) in each group were detected by Western blotting. Results:The immunofluorescence results showed that compared with the control group (1.00 ± 0.02), the fluorescence intensity of GSDMD in the fluoride, aluminum, and fluoride + aluminum groups (1.49 ± 0.02, 1.22 ± 0.04, 1.25 ± 0.03) were higher ( P < 0.05). The results of qRT-PCR showed that compared with the control group (1.00 ± 0.02, 1.00 ± 0.01, 1.00 ± 0.08, 1.00 ± 0.06), the mRNA expression levels of NLRP3 (1.21 ± 0.06, 1.60 ± 0.07, 1.42 ± 0.02), ASC (2.61 ± 0.07, 1.53 ± 0.01, 2.12 ± 0.03), Caspase-1 (1.32 ± 0.05, 1.53 ± 0.04, 2.07 ± 0.05), and GSDMD (1.60 ± 0.03, 1.65 ± 0.04, 2.23 ± 0.05) were higher in the fluoride, aluminum, and fluoride + aluminum groups ( P < 0.05). Western blotting results showed that compared with the control group (1.00 ± 0.04, 1.00 ± 0.08, 1.00 ± 0.05, 1.00 ± 0.02, 1.00 ± 0.03, 1.00 ± 0.06), the protein expression levels of NLRP3 (1.55 ± 0.06, 1.40 ± 0.07, 1.24 ± 0.05), ASC (1.66 ± 0.05, 1.48 ± 0.06, 1.32 ± 0.06), Caspase-1 (1.51 ± 0.02, 1.40 ± 0.01, 1.28 ± 0.03), GSDMD (1.24 ± 0.03, 1.31 ± 0.06, 1.18 ± 0.03), GSDMD-N (1.18 ± 0.04, 1.27 ± 0.03, 1.27 ± 0.03), and IL-1β (1.81 ± 0.03, 1.70 ± 0.08, 1.52 ± 0.05) were higher in the fluoride, aluminum, and fluoride + aluminum groups ( P < 0.05). Conclusion:Exposure to fluoride and aluminum alone and in combination can induce pyroptosis in NG108-15 cells, and the mechanism may be related to up-regulation of molecules related to the NLRP3/Caspase-1/GSDMD signaling pathway.

BACKGROUND:Metformin is currently considered the first-line medication for the treatment of type 2 diabetes.Metformin may delay the progression of osteoarthritis,but its specific mechanism of action remains unclear.OBJECTIVE:To evaluate the therapeutic effects and the related action mechanisms of metformin on osteoarthritis in rats.METHODS:(1)Network pharmacology:Potential common targets for metformin,osteoarthritis,and ferroptosis were screened using the CTD,SwissTargetPrediction,GeneCards,and OMIM databases.After importing the targets into the STRING database,protein-protein interaction analysis was conducted to identify the key targets for metformin,osteoarthritis,and ferroptosis.(2)Molecular docking:P53 and its downstream factor SLC7A11 protein structures in PDB format were downloaded from the PDB database.The 2D structure of metformin was converted to a 3D structure,and molecular docking of metformin with the proteins was performed using Discovery Studio 2019 Client.(3)In vivo experiments:Thirty male SD rats were randomly divided into three groups(n=10).The blank group did not receive surgery.The osteoarthritis model was established using the modified Hulth method for the model and metformin groups.One day after the surgery,rats in the metformin group were gavaged with metformin 200 mg/kg per day,while the blank and model groups were gavaged with physiological saline.Treatment continued for 4 weeks.Hematoxylin-eosin staining and Safranin O-fast green staining were used to observe the pathological morphology and structure of the knee cartilage,and Mankin scoring was performed.ELISA was used to measure the levels of tumor necrosis factor-α and interleukin-6 in the serum.The microplate method was used to measure serum ferroptosis-related indicators,including glutathione,malondialdehyde,and Fe2+.Immunofluorescence staining,western blot assay,and real-time qPCR were used to detect the protein and mRNA expression of P53,SLC7A11,glutathione peroxidase 4,proteoglycans,and matrix metalloproteinase 13 in the cartilage tissue of the rats.RESULTS AND CONCLUSION:(1)A total of 96 intersecting targets among metformin,osteoarthritis,and ferroptosis were identified.After protein-protein interaction analysis,77 potential targets were found.Further screening identified the core targets as TP53,AKT1,JUN,interleukin-6,MYC,interleukin-1β,and tumor necrosis factor-α,among others.(2)Docking analysis results showed that metformin bound strongly and stably with P53 and its downstream factor SLC7A11.(3)In the model group,the knee cartilage surface was irregular,with cartilage tissue defects and reduced chondrocyte numbers.Compared to the model group,the knee cartilage structure damage in the metformin group was significantly improved,with a smoother cartilage surface and increased chondrocyte numbers.The Mankin score in the model group was significantly higher than that in the blank group,while the Mankin score in the intervention group was significantly lower than that in the model group.(4)Compared with the model group,the metformin group had significantly lower levels of tumor necrosis factor-α,interleukin-6,malondialdehyde,and Fe2+,and significantly higher glutathione levels.(5)Compared to the model group,the metformin group had significantly increased protein and mRNA expression of SLC7A11,glutathione peroxidase 4,and proteoglycans,and significantly decreased protein and mRNA expression of P53 and matrix metalloproteinase 13 in their cartilage tissue.(6)The results indicate that metformin can effectively improve cartilage damage in osteoarthritis rats and alleviate chondrocyte ferroptosis by inhibiting the aberrantly activated P53/SLC7A11/glutathione peroxidase 4 signaling pathway.This improvement in chondrocyte iron metabolism and lipid peroxidation response further reduces cartilage matrix degradation and prevents further cartilage damage and inflammatory response.

Objective:To study the effects of fluoride and aluminum exposure alone and in combination on pyroptosis and the NOD-like receptor protein 3 (NLRP3)/cysteinyl aspartate specific proteinase-1 (Caspase-1)/gasdermin D (GSDMD) signaling pathway in NG108-15 cells.Methods:Using a factorial design method, NG108-15 cells cultured in vitro were divided into control group [0 mg/L sodium fluoride (NaF) + 0 mg/L aluminium trichloride (AlCl 3)], fluoride group (40 mg/L NaF + 0 mg/L AlCl 3), aluminum group (0 mg/L NaF + 160 mg/L AlCl 3), and fluoride + aluminum group (40 mg/L NaF + 160 mg/L AlCl 3) according to the concentrations of NaF and AlCl 3. After 24 hours of cultivation, the cells were collected for subsequent experiments. The fluorescence intensity of pyroptosis index GSDMD in each group was detected by immunofluorescence method. The mRNA expression levels of NLRP3, apoptosis associated speck-like protein (ASC), Caspase-1, and GSDMD in each group were detected by real-time fluorescence quantitative PCR (qRT-PCR). The protein expression levels of NLRP3, ASC, Caspase-1, GSDMD, gasdermin D N-terminus (GSDMD-N) and interleukin-1β (IL-1β) in each group were detected by Western blotting. Results:The immunofluorescence results showed that compared with the control group (1.00 ± 0.02), the fluorescence intensity of GSDMD in the fluoride, aluminum, and fluoride + aluminum groups (1.49 ± 0.02, 1.22 ± 0.04, 1.25 ± 0.03) were higher ( P < 0.05). The results of qRT-PCR showed that compared with the control group (1.00 ± 0.02, 1.00 ± 0.01, 1.00 ± 0.08, 1.00 ± 0.06), the mRNA expression levels of NLRP3 (1.21 ± 0.06, 1.60 ± 0.07, 1.42 ± 0.02), ASC (2.61 ± 0.07, 1.53 ± 0.01, 2.12 ± 0.03), Caspase-1 (1.32 ± 0.05, 1.53 ± 0.04, 2.07 ± 0.05), and GSDMD (1.60 ± 0.03, 1.65 ± 0.04, 2.23 ± 0.05) were higher in the fluoride, aluminum, and fluoride + aluminum groups ( P < 0.05). Western blotting results showed that compared with the control group (1.00 ± 0.04, 1.00 ± 0.08, 1.00 ± 0.05, 1.00 ± 0.02, 1.00 ± 0.03, 1.00 ± 0.06), the protein expression levels of NLRP3 (1.55 ± 0.06, 1.40 ± 0.07, 1.24 ± 0.05), ASC (1.66 ± 0.05, 1.48 ± 0.06, 1.32 ± 0.06), Caspase-1 (1.51 ± 0.02, 1.40 ± 0.01, 1.28 ± 0.03), GSDMD (1.24 ± 0.03, 1.31 ± 0.06, 1.18 ± 0.03), GSDMD-N (1.18 ± 0.04, 1.27 ± 0.03, 1.27 ± 0.03), and IL-1β (1.81 ± 0.03, 1.70 ± 0.08, 1.52 ± 0.05) were higher in the fluoride, aluminum, and fluoride + aluminum groups ( P < 0.05). Conclusion:Exposure to fluoride and aluminum alone and in combination can induce pyroptosis in NG108-15 cells, and the mechanism may be related to up-regulation of molecules related to the NLRP3/Caspase-1/GSDMD signaling pathway.

Objective:This study starts from the main problems existing in the current stage of the development of pediatric clinical research in China and focuses on the construction of a clinical research management system. By innovating the platform management model and enhancing personnel skills, we can provide support and assurance for improving the clinical research platform and its sustainable development in the future.Methods:This study established an integrated management platform for Industry-Sponsored Initiated Clinical Research Trial (IST) and Investigator Initiated Clinical Research (IIT) and implement integrated management of IST and IIT projects. At the same time, in response to the weak links in quality management, synchronous training of management and research talents should be implemented to achieve dual improvement in project management quality and platform service efficiency throughout the process.Results:We had optimized the clinical research organization structure, improved the management system, and strengthened the quality supervision of IIT projects by establishing rules and systems to improve the quality and efficiency of project management.Conclusions:Based on analyzing the current situation in China and referring to international experience, we have built a clinical research management platform in line with the development of the hospital, and implemented centralized management of the entire process of IST and IIT projects, strengthened project process quality control, which is expected to provide a reference for peers.

Objective:To summarize the clinical phenotype and genetic features of patients with relapsing encephalopathy with cerebellar ataxia (RECA) caused by ATP1A3 gene R756 variants. Methods:A retrospective analysis was performed on patients carrying the ATP1A3 gene R756 variants, identified by whole-exome sequencing of family members, at Capital Center for Children′s Health, Capital Medical University and Children's Medical Center, Peking University First Hospital from August 2005 to February 2024. Their clinical, laboratory, neuroimaging, electrophysiological and genetic characteristics were summarized. Results:A total of 13 RECA patients were enrolled in this study, including 8 males and 5 females. The age of onset was 8 months to 5 years, with a median age of onset of 18 months. All of 13 patients presented paroxysmal episodes of neurological decompensations triggered by fever and residual symptoms following the acute phase. During acute attack stage, ataxia was observed in all 13 cases, muscle weakness in 12 cases, dysarthria in 12 cases, altered consciousness in 10 cases, dysphagia in 10 cases, dystonic episodes in 4 cases, abnormal eye movement in 2 cases, choreoathetosis in 2 cases, and epileptic seizures in 1 case. All 13 patients had residual symptoms during the nonparoxysmal period, of whom 9 patients had ataxia, 9 patients had dysarthria, 4 patients had dystonia, 3 patients had cognitive disorders, and 1 patient had epileptic seizures. All 13 cases had ATP1A3 missense variants, and variant c.2266C>T/p.R756C was found in 6 cases, c.2267G>A/p.R756H in 5 cases, and c.2267G>T/p.R756L in 2 cases. Nine cases carried de novo variants, 4 with inherited variants. Conclusions:RECA caused by variants of ATP1A3 in residue 756 typically presents with an acute onset during infancy or early childhood, precipitated by febrile episodes and characterized by recurrent episodes of ataxia, with bulbar paralysis, muscle weakness and altered consciousness. Recurrence is common, and the most common persistent symptoms are cerebellar ataxia and dysarthria. A few patients have cognitive impairment. Three types of ATP1A3 gene variants R756C, R756H and R756L are related with RECA, and R756C is the most common variant.

Background: Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients. Methods: Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines. Results: Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher. Conclusion MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.

Background: s/Aims: Non-invasive models stratifying clinically significant portal hypertension (CSPH) are limited. Herein, we developed a new non-invasive model for predicting CSPH in patients with compensated cirrhosis and investigated whether carvedilol can prevent hepatic decompensation in patients with high-risk CSPH stratified using the new model. Methods: Non-invasive risk factors of CSPH were identified via systematic review and meta-analysis of studies involving patients with hepatic venous pressure gradient (HVPG). A new non-invasive model was validated for various performance aspects in three cohorts, i.e., a multicenter HVPG cohort, a follow-up cohort, and a carvediloltreating cohort. Results: In the meta-analysis with six studies (n=819), liver stiffness measurement and platelet count were identified as independent risk factors for CSPH and were used to develop the new “CSPH risk” model. In the HVPG cohort (n=151), the new model accurately predicted CSPH with cutoff values of 0 and –0.68 for ruling in and out CSPH, respectively. In the follow-up cohort (n=1,102), the cumulative incidences of decompensation events significantly differed using the cutoff values of <–0.68 (low-risk), –0.68 to 0 (medium-risk), and >0 (high-risk). In the carvediloltreated cohort, patients with high-risk CSPH treated with carvedilol (n=81) had lower rates of decompensation events than non-selective beta-blockers untreated patients with high-risk CSPH (n=613 before propensity score matching [PSM], n=162 after PSM). Conclusions Treatment with carvedilol significantly reduces the risk of hepatic decompensation in patients with high-risk CSPH stratified by the new model.

Background: Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients. Methods: Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines. Results: Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher. Conclusion MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.

Background: Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients. Methods: Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines. Results: Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher. Conclusion MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.