Objective:To establish and optimize a novel method, focus forming assay (FFA), for quantitation of influenza A virus (FluA) and compare its application performance with traditional plague forming assay (PFA).Methods:The foci chromogenic effects of three peroxidase substrates in immunostaining were compared. The PFA and FFA methods were used to explore FluA incubation times and plaque morphology on 12-well plates, and to determine optimal incubation times and virus adsorption volumes for different FluA subtypes on 96-well plates. The correlation between FFA and PFA was evaluated, and the optimized FFA was applied to the in vitro antiviral efficacy analysis of Favipiravir and neutralization test against different subtypes of FluA. Results:TRUEBLUE substrate was identified as the optimal substrate for foci visualization. Compared with the PFA, the FFA showed improved sensitivity and reduced detection time in FluA titration, and good correlation was shown between the two methods′ results. By replacing the 96-well plate with the 12-well plate for FFA titration of different subtypes of FluA, the detection time was shortened, and the amount of serum samples used could be further reduced by optimizing the virus adsorption volume. The half-maximal effective concentration of favipiravir against influenza viruses assessed by the FFA and PFA methods showed no significant difference, and was consistent with the results obtained from quantitative PCR. Additionally, the focus reduction neutralization test and hemagglutination inhibition assays demonstrated strong correlation in determining antibody titers against FluA in serum neutralization assays.Conclusions:The improved FFA method developed here provides a more efficient experimental tool for FluA titration, antiviral drug screening and broad-spectrum vaccine evaluation.

Objective To evaluate the sensitivity of swept-source optical coherence tomography(SS-OCT)in detecting early deminer-alization at the resin-enamel bonding interface,and the differences in enamel demineralization around restorations among different resins(Filtek Z350 XT,Filtek Bulk Fill,TetricN-Ceram Bulk Fill).Methods Twenty-seven extracted third molars were selected and prepared into 5-mm-thick specimens,and Class Ⅰ cavities measuring 3 mm × 3 mm × 4 mm were created on the occlusal surfaces.The specimens were randomly divided into three groups,with nine teeth in each group,and were respectively filled with Filtek Z350 XT(layered filling of 4 mm),Filtek Bulk Fill(bulk filling of 4 mm in one step),and Tetric N-Ceram Bulk Fill(bulk filling of 4 mm in one step).After applying acid-resistant nail varnish to non-experimental areas,the specimens were placed in a demineralizing solution for 4 weeks.SS-OCT and Micro-CT scans of the resin-enamel bonding interface were performed before demineralization and weekly thereafter to monitor the progression of demineralization and changes in demineralization depth were quantitatively analyzed.Results Both SS-OCT and Micro-CT were capable of non-destructive dynamic monitoring of demineralization at the resin-enamel bonding inter-face.After demineralization,four types of patterns were observed at the resin-enamel bonding interface.At different stages of demineral-ization,no significant differences in demineralization depth were detected among the three resins using either SS-OCT or Micro-CT.There was a high level of agreement between the demineralization depth measurements obtained from SS-OCT and Micro-CT at each demineralization stage(ICC:0.760-0.897).Conclusion The early demineralization of resin-enamelbonding interface can be noninva-sively detected by SS-OCT,and there was no significant differenceamong three resins in their ability to resist enamel demineralization around the restoration.

Liposomes serve as critical carriers for drugs and vaccines, with their biological effects influenced by their size. The microfluidic method, renowned for its precise control, reproducibility, and scalability, has been widely employed for liposome preparation. Although some studies have explored factors affecting liposomal size in microfluidic processes, most focus on small-sized liposomes, predominantly through experimental data analysis. However, the production of larger liposomes, which are equally significant, remains underexplored. In this work, we thoroughly investigate multiple variables influencing liposome size during microfluidic preparation and develop a machine learning (ML) model capable of accurately predicting liposomal size. Experimental validation was conducted using a staggered herringbone micromixer (SHM) chip. Our findings reveal that most investigated variables significantly influence liposomal size, often interrelating in complex ways. We evaluated the predictive performance of several widely-used ML algorithms, including ensemble methods, through cross-validation (CV) for both liposome size and polydispersity index (PDI). A standalone dataset was experimentally validated to assess the accuracy of the ML predictions, with results indicating that ensemble algorithms provided the most reliable predictions. Specifically, gradient boosting was selected for size prediction, while random forest was employed for PDI prediction. We successfully produced uniform large (600 nm) and small (100 nm) liposomes using the optimised experimental conditions derived from the ML models. In conclusion, this study presents a robust methodology that enables precise control over liposome size distribution, offering valuable insights for medicinal research applications.

Objective To investigate the effects of Duzhi Pills combined with MOTOmed training on limb function and activities of daily living in patients with cerebral infarction of qi deficiency and blood stasis type.Methods A prospective randomized controlled trial enrolled 98 cerebral infarction patients with qi deficiency and blood stasis treated at Qinhuangdao Hospital,Oriental Hospital of Beijing University of Chinese Medicine(Qinhuangdao Hospital of Traditional Chinese Medicine)from June 2022 to December 2024.Participants were randomized into control group(n=49,receiving conventional rehabilitation)and observation group(n=49,receiving additional Duzhi Pills+MOTOmed training)for 8 weeks.All patients received standard medication.Limb motor function and activities of daily living,and quality of life were assessed before and after treatment.Results(1)After treatment,both groups showed significant increases in Carroll Hand Function Test for Upper Extremity(CHFT-UE)scores,Fugl-Meyer Assessment Scale for Upper Extremity(FMA-UE)scores,and Simple Test for Evaluating Hand Function(STEF)scores compared to baseline(P＜0.05),with significantly greater improvements observed in the observation group(P＜0.01).(2)After treatment,both groups exhibited significant increases in Barthel Index scores for daily activity(P＜0.05),with the observation group demonstrating significantly greater improvement(P＜0.01).(3)After treatment,all subscale scores of the 36-Item Short-Form Health Survey(SF-36),including physical function,bodily pain,mental health,vitality,social functioning,emotional role,and general health,were significantly increased in both groups(P＜0.05),with significantly greater improvements in the observation group,the difference being statistically significant(P＜0.01).Conclusion The combined regimen of Duzhi Pills and MOTOmed training significantly enhances limb function,daily activity,and quality of life in cerebral infarction patients with qi deficiency and blood stasis syndrome,demonstrating clinical value for rehabilitation.

Liposomes serve as critical carriers for drugs and vaccines,with their biological effects influenced by their size.The microfluidic method,renowned for its precise control,reproducibility,and scalability,has been widely employed for liposome preparation.Although some studies have explored factors affecting liposomal size in microfluidic processes,most focus on small-sized liposomes,predominantly through experimental data analysis.However,the production of larger liposomes,which are equally significant,remains underexplored.In this work,we thoroughly investigate multiple variables influencing liposome size during microfluidic preparation and develop a machine learning(ML)model capable of accurately predicting liposomal size.Experimental validation was conducted using a staggered herringbone micromixer(SHM)chip.Our findings reveal that most investigated variables significantly influence liposomal size,often interrelating in complex ways.We evaluated the predictive performance of several widely-used ML algorithms,including ensemble methods,through cross-validation(CV)for both lipo-some size and polydispersity index(PDI).A standalone dataset was experimentally validated to assess the accuracy of the ML predictions,with results indicating that ensemble algorithms provided the most reliable predictions.Specifically,gradient boosting was selected for size prediction,while random forest was employed for PDI prediction.We successfully produced uniform large(600 nm)and small(100 nm)liposomes using the optimised experimental conditions derived from the ML models.In conclusion,this study presents a robust methodology that enables precise control over liposome size distribution,of-fering valuable insights for medicinal research applications.

Objective To compare the adverse reactions to pegasparaginase(PEG-ASP)and Erwiniaasparaginase(ASP)in the treat-ment of pediatric acute lymphoblastic leukemia(ALL)and to provide clinical guidance for treatment.Methods A total of 13 patients who were switched from PEG-ASP to Erwinia ASP due to allergic reactions during the induction phase of ALL treatment at the Depart-ment of Hematology and Oncology,Children's Hospital Affiliated of Zhengzhou University between February 2022 and February 2023 were selected as the case group.A control group of 26 patients treated with PEG-ASP and matched for sex at a ratio of 1∶2 was also selected.Pre-and post-treatment coagulation function,blood biochemistry,blood ammonia level,and gastrointestinal reactions were compared between the groups.Results There were no significant differences in AST,TB,UB,TC,TG,AMY,UN,Cr,APTT,and Fbg levels between the two groups,pre-and post-treatment.However,ALT,GGT,BB,and Glu levels were higher in the control group,whereas albumin was lower than in the case group(P<0.05).Both groups had an increased post-treatment blood ammonia level(P<0.05),with significantly a higher level observed in the case group than in the control group(P<0.001).No significant differences in adverse gastro-intestinal reactions were observed between the two groups.Conclusion Erwinia ASP and PEG-ASP affected liver function in pediatric ALL re-induction therapy with minimal effects on coagulation and weaker gastrointestinal reaction.Erwinia ASP significantly affected the blood ammonia level,and both treatments caused mild gastrointestinal reactions.Regular blood ammonia monitoring,post Erwinia ASP treatment,is recommended.

Background::Hepatitis B poses a heavy burden for children in China, however, the national studies on the distributional characteristics and health care costs of children with severe hepatitis B is still lacking. This study aimed to analyze the disease characteristics, health economic effects, and medical cost for children with severe hepatitis B in China.Methods::Based on patient information in the Hospital Quality Monitoring System, cases with severe hepatitis B were divided into four groups according to age, and the etiology and symptoms of each group were quantified. The cost of hospitalization was calculated for cases with different disease processes, and severity of disease. The spatial aggregation of cases and the relationship with health economic factors were analyzed by Moran’s I analysis. Results::The total number of children discharged with hepatitis B from January 2016 to April 2022 was 1603, with an average age of 10.5 years. Liver failure cases accounted for 43.48% (697/1603) of total cases and cirrhosis cases accounted for 11.23% (180/1603). According to the grouping of disease progression, there were 1292 cases without associated complications, and the median hospitalization cost was $818.12. According to the spatial analysis, the aggregation of cases was statistically significant at the prefectural and provincial levels in 2019, 2020, and 2021 (all P <0.05). The number of severe cases was negatively correlated with gross domestic product (Moran’s I <0) and percentage of urban population (Moran’s I <0), and positively correlated with the number of pediatric beds per million population (Moran’s I >0). Conclusion::The number of severe hepatitis B cases is low in areas with high gross domestic product levels and high urban population ratios, and health care costs have been declining over the years.

Objective To compare the adverse reactions to pegasparaginase(PEG-ASP)and Erwiniaasparaginase(ASP)in the treat-ment of pediatric acute lymphoblastic leukemia(ALL)and to provide clinical guidance for treatment.Methods A total of 13 patients who were switched from PEG-ASP to Erwinia ASP due to allergic reactions during the induction phase of ALL treatment at the Depart-ment of Hematology and Oncology,Children's Hospital Affiliated of Zhengzhou University between February 2022 and February 2023 were selected as the case group.A control group of 26 patients treated with PEG-ASP and matched for sex at a ratio of 1∶2 was also selected.Pre-and post-treatment coagulation function,blood biochemistry,blood ammonia level,and gastrointestinal reactions were compared between the groups.Results There were no significant differences in AST,TB,UB,TC,TG,AMY,UN,Cr,APTT,and Fbg levels between the two groups,pre-and post-treatment.However,ALT,GGT,BB,and Glu levels were higher in the control group,whereas albumin was lower than in the case group(P<0.05).Both groups had an increased post-treatment blood ammonia level(P<0.05),with significantly a higher level observed in the case group than in the control group(P<0.001).No significant differences in adverse gastro-intestinal reactions were observed between the two groups.Conclusion Erwinia ASP and PEG-ASP affected liver function in pediatric ALL re-induction therapy with minimal effects on coagulation and weaker gastrointestinal reaction.Erwinia ASP significantly affected the blood ammonia level,and both treatments caused mild gastrointestinal reactions.Regular blood ammonia monitoring,post Erwinia ASP treatment,is recommended.

Objective To evaluate the sensitivity of swept-source optical coherence tomography(SS-OCT)in detecting early deminer-alization at the resin-enamel bonding interface,and the differences in enamel demineralization around restorations among different resins(Filtek Z350 XT,Filtek Bulk Fill,TetricN-Ceram Bulk Fill).Methods Twenty-seven extracted third molars were selected and prepared into 5-mm-thick specimens,and Class Ⅰ cavities measuring 3 mm × 3 mm × 4 mm were created on the occlusal surfaces.The specimens were randomly divided into three groups,with nine teeth in each group,and were respectively filled with Filtek Z350 XT(layered filling of 4 mm),Filtek Bulk Fill(bulk filling of 4 mm in one step),and Tetric N-Ceram Bulk Fill(bulk filling of 4 mm in one step).After applying acid-resistant nail varnish to non-experimental areas,the specimens were placed in a demineralizing solution for 4 weeks.SS-OCT and Micro-CT scans of the resin-enamel bonding interface were performed before demineralization and weekly thereafter to monitor the progression of demineralization and changes in demineralization depth were quantitatively analyzed.Results Both SS-OCT and Micro-CT were capable of non-destructive dynamic monitoring of demineralization at the resin-enamel bonding inter-face.After demineralization,four types of patterns were observed at the resin-enamel bonding interface.At different stages of demineral-ization,no significant differences in demineralization depth were detected among the three resins using either SS-OCT or Micro-CT.There was a high level of agreement between the demineralization depth measurements obtained from SS-OCT and Micro-CT at each demineralization stage(ICC:0.760-0.897).Conclusion The early demineralization of resin-enamelbonding interface can be noninva-sively detected by SS-OCT,and there was no significant differenceamong three resins in their ability to resist enamel demineralization around the restoration.

Objective:To establish and optimize a novel method, focus forming assay (FFA), for quantitation of influenza A virus (FluA) and compare its application performance with traditional plague forming assay (PFA).Methods:The foci chromogenic effects of three peroxidase substrates in immunostaining were compared. The PFA and FFA methods were used to explore FluA incubation times and plaque morphology on 12-well plates, and to determine optimal incubation times and virus adsorption volumes for different FluA subtypes on 96-well plates. The correlation between FFA and PFA was evaluated, and the optimized FFA was applied to the in vitro antiviral efficacy analysis of Favipiravir and neutralization test against different subtypes of FluA. Results:TRUEBLUE substrate was identified as the optimal substrate for foci visualization. Compared with the PFA, the FFA showed improved sensitivity and reduced detection time in FluA titration, and good correlation was shown between the two methods′ results. By replacing the 96-well plate with the 12-well plate for FFA titration of different subtypes of FluA, the detection time was shortened, and the amount of serum samples used could be further reduced by optimizing the virus adsorption volume. The half-maximal effective concentration of favipiravir against influenza viruses assessed by the FFA and PFA methods showed no significant difference, and was consistent with the results obtained from quantitative PCR. Additionally, the focus reduction neutralization test and hemagglutination inhibition assays demonstrated strong correlation in determining antibody titers against FluA in serum neutralization assays.Conclusions:The improved FFA method developed here provides a more efficient experimental tool for FluA titration, antiviral drug screening and broad-spectrum vaccine evaluation.