AIM:To assess the influence of preoperative lacrimal passage probing and dilation on the therapeutic effectiveness of nasolacrimal silicone intubation in cases of incomplete primary acquired nasolacrimal duct obstruction(PANDO).METHODS: A retrospective case-control clinical study was conducted. The study included patients with incomplete PANDO who underwent nasolacrimal silicone intubation at the hospital between October 2022 and October 2024. Participants were categorized into two groups based on their receipt of nasolacrimal duct probing and dilation prior to intubation. Group A consisted of patients who had undergone one or more sessions of nasolacrimal duct probing and dilation before nasolacrimal silicone intubation, while Group B included patients who did not receive nasolacrimal duct probing and dilation before intubation. All patients had their nasolacrimal silicone tubes removed 3 mo postoperatively and were monitored for 6 mo following tube removal to evaluate surgical success rates.RESULTS: A total of 136 patients(152 eyes)completed the six-month follow-up period. The overall surgical success rate for both groups was 72.4%(110/152 eyes).Group A included 22 males and 36 females, mean age 58.09±11.07 y, and achieved a success rate of 82.8%(53/64 eyes), while Group B included 27 males and 51 females, mean age 59.35±8.84 y and attained a success rate of 64.8%(57/88 eyes). The difference in surgical success rates between the two groups was statistically significant(P=0.014). Furthermore, Group A achieved a complete success rate of 67.2%(43/64 eyes), compared to 47.7%(42/88 eyes)in Group B, with this difference also reaching statistical significance(P=0.017). No serious postoperative complications were observed in either group.CONCLUSION: Nasolacrimal silicone intubation is a safe and effective intervention for incomplete PANDO, with preoperative nasolacrimal duct probing and dilation potentially enhancing therapeutic outcomes.

In recent years,the number of Chest Pain Centers in China has increased rapidly.As of December 2024,there were 5 947 registered units of Chest Pain Centers nationwide,2 994 of which had passed the acceptance.As of April 2025,18.34 million acute chest pain cases have been treated at Chest Pain Centers nationwide,according to the Chest Pain Center reporting platform.The Chest Pain Centers treated more than 3.62 million patients with acute chest pain in 2024,including more than 770 000 patients with acute myocardial infarction,and achieved positive results:significantly increasing the proportion of reperfusion therapy for acute ST-segment elevation myocardial infarction patients,ensuring the treatment of myocardial infarction patients in a short time,shortening the length of patient hospitalization,and reducing the hospitalization costs and mortality of patients.Through the construction of Chest Pain Center,the treatment process was significantly optimized and the prognosis of patients was improved.At present,the construction and quality control of Chest Pain Centers still need to be further promoted.In some areas,the construction proportion of Chest Pain Centers and the level of diagnosis and treatment of chest pain need to be improved,patient delay is still an important challenge for the treatment of acute chest pain in China.

Objective:To establish and evaluate a second-tier screening method for neonatal congenital adrenal hyperplasia (CAH) and develop appropriate screening interpretation criteria.Methods:We employed liquid chromatography-tandem mass spectrometry to simultaneously detect five steroid hormones in dried blood spots: 17α-hydroxyprogesterone (17α-OHP), androstenedione (A4), 11-deoxycortisol (11-DOC), 21-deoxycortisol (21-DOC), and cortisol (F), calculating (17α-OHP+A4)/F and (17α-OHP+21-DOC)/F ratios for second-tier CAH screening. The study utilized 429 residual dried blood spot samples from neonates (0-7 days) who completed first-tier screening between January 2020 and March 2024 in Guangzhou Women and Children's Medical Center, Guangzhou Medical University, including first-tier negatives ( n=369), confirmed false positives ( n=50), and CYP21A2-confirmed 21-hydroxylase deficiency patients ( n=10). Mann-Whitney U and Kruskal-Wallis tests analyzed steroid concentration variations across gestational ages and birth weights in all negative samples, with reference intervals established via P2.5- P97.5 percentiles and screening cutoffs set at population P97.5. Receiver operating characteristic (ROC) curve analysis identified optimal interpretation indicators among steroid hormone profiles, with second-tier screening performance evaluated by comparing sensitivity and specificity across different steroid hormone indicators to establish the optimal diagnostic criteria. Results:The five steroid hormones demonstrated intra-assay precision with coefficient of variation (CV) of 9.8%-14.2% and inter-assay precision with CV of 4.7%-14.4% across three different concentration levels of quality control materials. Accuracy ranged from 98.5% to 110.0% and the lower limits of quantification were 0.25 ng/ml for 17α-OHP, 0.05 ng/ml for A4/11-DOC, 0.31 ng/ml for 21-DOC, and 0.1 ng/ml for F. Stratification by gestational age categorized 17α-OHP into ≤31, 32-34, and ≥35 weeks; A4 into ≤31, 32-36, and ≥37 weeks; and 11-DOC into ≤31 and ≥32 weeks, while the remaining indicators were not stratified. When grouped by birth weight (low/normal), all measured parameters except 21-DOC showed statistically significant differences between groups (all P<0.05). Established reference intervals included 17α-OHP: 0.53-7.82 ng/ml (≤31 weeks), <0.25-3.60 ng/ml (32-34 weeks), <0.25-1.64 ng/ml (≥35 weeks); A4: 0.12-2.36 ng/ml (≤31 weeks), <0.05-1.45 ng/ml (32-36 weeks), 0.17-0.95 ng/ml (≥37 weeks); 11-DOC: 0.43-4.04 ng/ml (≤31 weeks), 0.08-1.46 ng/ml (≥32 weeks); F: 1.70-83.70 ng/ml; 21-DOC: <0.31-0.69 ng/ml; (17α-OHP+A4)/F: 0.01-0.74; and (17α-OHP+21-DOC)/F: 0.01-0.69. Comprehensive comparison of CAH second-tier screening performance demonstrated that interpretation based on elevated 17α-OHP accompanied by either elevated 21-DOC or elevated ratios [(17α-OHP+A4)/F or (17α-OHP+21-DOC)/F] achieved 100% sensitivity, 96% specificity, and a 96% reduction in false-positive rate. Conclusion:The application of liquid chromatography-tandem mass spectrometry for multi-steroid hormone profiling in second-tier neonatal CAH screening, utilizing gestational age-specific 17α-OHP cutoffs combined with elevated 21-DOC or ratio criteria, demonstrated 100% screening sensitivity while substantially reducing false-positive rates from primary screening, though further validation with expanded sample sizes remains necessary.

This study was aimed at investigating the effects of demethylase fat mass and obesity-associated protein(FTO)and methyltransferase methyltransferase like protein 3(METTL3),key molecules in N6-methyladenosine(m6A)modification,on the replication and proliferation of Japanese encephalitis virus(JEV).Recombinant lentiviruses were generated by packaging the FTO and green fluorescent protein into lentiviral vectors.Neuro2a cells,a mouse neuroblastoma cell line,were infected with the lentivirus,and stable FTO-expressing cell lines were obtained through puromycin selection.Successful overexpression of FTO was confirmed through fluorescence microscopy,real-time quantitative PCR,and western blot analysis.When Neuro2a cells overexpressing FTO were infected with JEV,the overexpression of FTO decreased JEV replication in the cells,and increased the expression of interferon(IFN)and related molecules.Additionally,treatment of JEV-infected Neuro2a cells with the METTL3-specific inhibitor STM2457 resulted in a dose-dependent decrease in JEV replication and viral protein expression.These findings suggested that lowering m6A methylation levels inhibits JEV replication,thus shedding light on the regulatory role of methylation modification in JEV replication.

In recent years,the number of Chest Pain Centers in China has increased rapidly.As of December 2024,there were 5 947 registered units of Chest Pain Centers nationwide,2 994 of which had passed the acceptance.As of April 2025,18.34 million acute chest pain cases have been treated at Chest Pain Centers nationwide,according to the Chest Pain Center reporting platform.The Chest Pain Centers treated more than 3.62 million patients with acute chest pain in 2024,including more than 770 000 patients with acute myocardial infarction,and achieved positive results:significantly increasing the proportion of reperfusion therapy for acute ST-segment elevation myocardial infarction patients,ensuring the treatment of myocardial infarction patients in a short time,shortening the length of patient hospitalization,and reducing the hospitalization costs and mortality of patients.Through the construction of Chest Pain Center,the treatment process was significantly optimized and the prognosis of patients was improved.At present,the construction and quality control of Chest Pain Centers still need to be further promoted.In some areas,the construction proportion of Chest Pain Centers and the level of diagnosis and treatment of chest pain need to be improved,patient delay is still an important challenge for the treatment of acute chest pain in China.

Currently,there are practical and technical difficulties in the construction of clinical ques-tions in the development of clinical practice guidelines.Clinicians or guideline developers seldom construct clin-ical questions based the actual case scenario,leading to some information loss between structured and actual clinical connotation.To overcome this challenge,we proposed a case-guided questions construction approach,and carried out case research and verification in the formulation of the guideline.We found that this method could more efficiently and scientifically assist the formulation of clinical questions,and provide reference for clinicians or guideline developers.

AIM To investigate the effects of Xuesaitong Capsules(Panax notoginseng saponins)on ischemia/reperfusion injury in a mouse model of skin frostbite.METHODS The mice were randomly divided into the control group,the model group,the dexamethasone group(1 mg/kg),and the low-dose,medium-dose,and high-dose Xuesaitong Capsules groups(0.036,0.072,and 0.144 g/kg),with eight mice in each group.A frostbite model was established using a dry ice-cooled ceramic(ferrite)magnet.On the 2nd day after modeling,each group started its corresponding dosing by gavage for 14 consecutive days.The wound healing,histopathological changes,and serum levels of high-sensitivity C-reactive protein(hs-CRP),thromboxane B2(TXB2),6-keto-prostaglandin F1α(6-K-PGF1α),nitric oxide(NO)and endothelin(ET)were assessed using ELISA.The superoxide dismutase(SOD)activity and malondialdehyde(MDA)levels in skin tissues were measured biochemically.The protein expressions of tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6,Toll-like receptor(TLR)4 and phosphorylated nuclear factor-KB p65(p-NF-κB p65)in skin tissues were determined by Western blot.Additionally,LncRNA H19 mRNA expression in skin tissues was evaluated using RT-qPCR.RESULTS After the final administration,compared with the control group,the model group exhibited partial scab detachment,wound healing,and larger wound areas;hyperkeratosis with incomplete keratinization,detachment of the dermis and subcutaneous tissue,partial loss of appendages,subcutaneous edema,and dilated,congested,and hemorrhagic stromal vessels with extensive lymphocyte infiltration revealed by the histopathological examination;elevated serum levels of hs-CRP,TXB2,and ET(P＜0.05,P＜0.01);decreased 6-K-PGF1α and NO levels(P＜0.05,P＜0.01);reduced SOD activity in skin tissues(P＜0.01);increased MDA levels(P＜0.01);and upregulated protein expressions of TNF-α,IL-1β,IL-6,TLR4 and p-NF-κB p65,as well as LncRNA H19 mRNA expression(P＜0.05,P＜0.01).Compared with the model group,the group intervened with high-dose Xuesaitong Capsules displayed reduced wound areas(P＜0.01);decreased serum levels of hs-CRP,TXB2 and ET(P＜0.05,P＜0.01);increased 6-K-PGF1α and NO levels(P＜0.05,P＜0.01);enhanced SOD activity(P＜0.05,P＜0.01);reduced MDA level in skin tissues(P＜0.05,P＜0.01);and down-regulated TNF-α,IL-1β,IL-6,TLR4 and p-NF-κB p65 protein expressions and suppressed LncRNA H19 mRNA expression in skin tissues as well(P＜0.05,P＜0.01).CONCLUSION Xuesaitong Capsules alleviate ischemia/reperfusion injury in frostbite-injured mice by exerting anti-inflammatory and anti oxidative stress effects and restoring vascular endothelial function mediated by the downregulation of LncRNA H19 expression and inhibition of the TLR4/NF-κB signaling pathway.

This study was aimed at investigating the effects of demethylase fat mass and obesity-associated protein(FTO)and methyltransferase methyltransferase like protein 3(METTL3),key molecules in N6-methyladenosine(m6A)modification,on the replication and proliferation of Japanese encephalitis virus(JEV).Recombinant lentiviruses were generated by packaging the FTO and green fluorescent protein into lentiviral vectors.Neuro2a cells,a mouse neuroblastoma cell line,were infected with the lentivirus,and stable FTO-expressing cell lines were obtained through puromycin selection.Successful overexpression of FTO was confirmed through fluorescence microscopy,real-time quantitative PCR,and western blot analysis.When Neuro2a cells overexpressing FTO were infected with JEV,the overexpression of FTO decreased JEV replication in the cells,and increased the expression of interferon(IFN)and related molecules.Additionally,treatment of JEV-infected Neuro2a cells with the METTL3-specific inhibitor STM2457 resulted in a dose-dependent decrease in JEV replication and viral protein expression.These findings suggested that lowering m6A methylation levels inhibits JEV replication,thus shedding light on the regulatory role of methylation modification in JEV replication.

AIM To investigate the effects of Xuesaitong Capsules(Panax notoginseng saponins)on ischemia/reperfusion injury in a mouse model of skin frostbite.METHODS The mice were randomly divided into the control group,the model group,the dexamethasone group(1 mg/kg),and the low-dose,medium-dose,and high-dose Xuesaitong Capsules groups(0.036,0.072,and 0.144 g/kg),with eight mice in each group.A frostbite model was established using a dry ice-cooled ceramic(ferrite)magnet.On the 2nd day after modeling,each group started its corresponding dosing by gavage for 14 consecutive days.The wound healing,histopathological changes,and serum levels of high-sensitivity C-reactive protein(hs-CRP),thromboxane B2(TXB2),6-keto-prostaglandin F1α(6-K-PGF1α),nitric oxide(NO)and endothelin(ET)were assessed using ELISA.The superoxide dismutase(SOD)activity and malondialdehyde(MDA)levels in skin tissues were measured biochemically.The protein expressions of tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6,Toll-like receptor(TLR)4 and phosphorylated nuclear factor-KB p65(p-NF-κB p65)in skin tissues were determined by Western blot.Additionally,LncRNA H19 mRNA expression in skin tissues was evaluated using RT-qPCR.RESULTS After the final administration,compared with the control group,the model group exhibited partial scab detachment,wound healing,and larger wound areas;hyperkeratosis with incomplete keratinization,detachment of the dermis and subcutaneous tissue,partial loss of appendages,subcutaneous edema,and dilated,congested,and hemorrhagic stromal vessels with extensive lymphocyte infiltration revealed by the histopathological examination;elevated serum levels of hs-CRP,TXB2,and ET(P＜0.05,P＜0.01);decreased 6-K-PGF1α and NO levels(P＜0.05,P＜0.01);reduced SOD activity in skin tissues(P＜0.01);increased MDA levels(P＜0.01);and upregulated protein expressions of TNF-α,IL-1β,IL-6,TLR4 and p-NF-κB p65,as well as LncRNA H19 mRNA expression(P＜0.05,P＜0.01).Compared with the model group,the group intervened with high-dose Xuesaitong Capsules displayed reduced wound areas(P＜0.01);decreased serum levels of hs-CRP,TXB2 and ET(P＜0.05,P＜0.01);increased 6-K-PGF1α and NO levels(P＜0.05,P＜0.01);enhanced SOD activity(P＜0.05,P＜0.01);reduced MDA level in skin tissues(P＜0.05,P＜0.01);and down-regulated TNF-α,IL-1β,IL-6,TLR4 and p-NF-κB p65 protein expressions and suppressed LncRNA H19 mRNA expression in skin tissues as well(P＜0.05,P＜0.01).CONCLUSION Xuesaitong Capsules alleviate ischemia/reperfusion injury in frostbite-injured mice by exerting anti-inflammatory and anti oxidative stress effects and restoring vascular endothelial function mediated by the downregulation of LncRNA H19 expression and inhibition of the TLR4/NF-κB signaling pathway.

Objective:To establish and evaluate a second-tier screening method for neonatal congenital adrenal hyperplasia (CAH) and develop appropriate screening interpretation criteria.Methods:We employed liquid chromatography-tandem mass spectrometry to simultaneously detect five steroid hormones in dried blood spots: 17α-hydroxyprogesterone (17α-OHP), androstenedione (A4), 11-deoxycortisol (11-DOC), 21-deoxycortisol (21-DOC), and cortisol (F), calculating (17α-OHP+A4)/F and (17α-OHP+21-DOC)/F ratios for second-tier CAH screening. The study utilized 429 residual dried blood spot samples from neonates (0-7 days) who completed first-tier screening between January 2020 and March 2024 in Guangzhou Women and Children's Medical Center, Guangzhou Medical University, including first-tier negatives ( n=369), confirmed false positives ( n=50), and CYP21A2-confirmed 21-hydroxylase deficiency patients ( n=10). Mann-Whitney U and Kruskal-Wallis tests analyzed steroid concentration variations across gestational ages and birth weights in all negative samples, with reference intervals established via P2.5- P97.5 percentiles and screening cutoffs set at population P97.5. Receiver operating characteristic (ROC) curve analysis identified optimal interpretation indicators among steroid hormone profiles, with second-tier screening performance evaluated by comparing sensitivity and specificity across different steroid hormone indicators to establish the optimal diagnostic criteria. Results:The five steroid hormones demonstrated intra-assay precision with coefficient of variation (CV) of 9.8%-14.2% and inter-assay precision with CV of 4.7%-14.4% across three different concentration levels of quality control materials. Accuracy ranged from 98.5% to 110.0% and the lower limits of quantification were 0.25 ng/ml for 17α-OHP, 0.05 ng/ml for A4/11-DOC, 0.31 ng/ml for 21-DOC, and 0.1 ng/ml for F. Stratification by gestational age categorized 17α-OHP into ≤31, 32-34, and ≥35 weeks; A4 into ≤31, 32-36, and ≥37 weeks; and 11-DOC into ≤31 and ≥32 weeks, while the remaining indicators were not stratified. When grouped by birth weight (low/normal), all measured parameters except 21-DOC showed statistically significant differences between groups (all P<0.05). Established reference intervals included 17α-OHP: 0.53-7.82 ng/ml (≤31 weeks), <0.25-3.60 ng/ml (32-34 weeks), <0.25-1.64 ng/ml (≥35 weeks); A4: 0.12-2.36 ng/ml (≤31 weeks), <0.05-1.45 ng/ml (32-36 weeks), 0.17-0.95 ng/ml (≥37 weeks); 11-DOC: 0.43-4.04 ng/ml (≤31 weeks), 0.08-1.46 ng/ml (≥32 weeks); F: 1.70-83.70 ng/ml; 21-DOC: <0.31-0.69 ng/ml; (17α-OHP+A4)/F: 0.01-0.74; and (17α-OHP+21-DOC)/F: 0.01-0.69. Comprehensive comparison of CAH second-tier screening performance demonstrated that interpretation based on elevated 17α-OHP accompanied by either elevated 21-DOC or elevated ratios [(17α-OHP+A4)/F or (17α-OHP+21-DOC)/F] achieved 100% sensitivity, 96% specificity, and a 96% reduction in false-positive rate. Conclusion:The application of liquid chromatography-tandem mass spectrometry for multi-steroid hormone profiling in second-tier neonatal CAH screening, utilizing gestational age-specific 17α-OHP cutoffs combined with elevated 21-DOC or ratio criteria, demonstrated 100% screening sensitivity while substantially reducing false-positive rates from primary screening, though further validation with expanded sample sizes remains necessary.