ObjectiveTo optimize the extraction method for polysaccharides from turnip（Brassica rapa）， and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction（BP-1） and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index， a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio， extraction time and extraction temperature， in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry（UV）， gas chromatography-mass spectrometry（GC-MS）， Congo red assay， and Fourier-transform infrared spectroscopy（FT-IR） to determine purity， monosaccharide composition， triple-helix structure， and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy（SEM） and atomic force microscopy（AFM）. Zebrafish were divided into the blank group（adding E3 medium）， and BP-1-1， BP-1-10， BP-1-50， BP-1-200， BP-1-1 000 groups（adding BP-1 solutions at concentrations of 1， 10， 50， 200， 1 000 mg·L^-1， respectively）， and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype， survival rate， malformation rate， and heart rate. Experimental animals were randomly divided into the blank group， model group， BP-1-10 group（10 mg·L^-1）， BP-1-50 group（50 mg·L^-1）， and BP-1-200 group（200 mg·L^-1）. The blank group was cultured in E3 medium， the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization（dpf） zebrafish embryos using 10 g·L^-1 of glucose combined with 500 µmol·L^-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution， while the blank and model groups received an equal volume of saline. Glucose and insulin（INS） levels were measured using enzyme-linked immunosorbent assay（ELISA） kits， the effects on the liver were observed by hematoxylin-eosin（HE） histopathological sections. The mRNA expression levels of glucagon（Glucagon）， insulin（Insa）， and phosphoenolpyruvate carboxykinase 1（PCK1） were detected with real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultsThe optimized extraction conditions were determined as follows：solid-to-liquid ratio of 1∶40（g·mL^-1）， extraction time of 66 min， and extraction temperature of 79 ℃. Under these conditions， the yield of turnip polysaccharides was （10.34±0.96）%. UV analysis indicated that BP-1 contained no proteins or nucleic acids， GC-MS analysis revealed that BP-1 consisted of six monosaccharides（arabinose， rhamnose， ribose， mannose， galactose and glucose）. Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure， FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids， SEM analysis revealed an irregular flaky structure with a flat and smooth surface， AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L^-1. Pharmacodynamic results showed that， compared with the blank group， the model group exhibited significantly increased glucose levels and significantly decreased INS levels（P<0.01）. Compared with the model group， the BP-1-50 group significantly reduced glucose levels and increased INS levels（P<0.05）. Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal， with hepatocytes appearing plump. Real-time PCR results showed that， compared with the blank group， the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1， and significantly downregulated mRNA expression of Insa（P<0.01）. Compared with the model group， the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1， and significantly upregulated mRNA expression of Insa（P<0.01）. ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate， is simple to operate， has low costs， making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides， contains α-glycosidic bonds and uronic acids， exhibits hypoglycemic activity， and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.

ObjectiveTo optimize the extraction method for polysaccharides from turnip（Brassica rapa）， and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction（BP-1） and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index， a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio， extraction time and extraction temperature， in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry（UV）， gas chromatography-mass spectrometry（GC-MS）， Congo red assay， and Fourier-transform infrared spectroscopy（FT-IR） to determine purity， monosaccharide composition， triple-helix structure， and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy（SEM） and atomic force microscopy（AFM）. Zebrafish were divided into the blank group（adding E3 medium）， and BP-1-1， BP-1-10， BP-1-50， BP-1-200， BP-1-1 000 groups（adding BP-1 solutions at concentrations of 1， 10， 50， 200， 1 000 mg·L^-1， respectively）， and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype， survival rate， malformation rate， and heart rate. Experimental animals were randomly divided into the blank group， model group， BP-1-10 group（10 mg·L^-1）， BP-1-50 group（50 mg·L^-1）， and BP-1-200 group（200 mg·L^-1）. The blank group was cultured in E3 medium， the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization（dpf） zebrafish embryos using 10 g·L^-1 of glucose combined with 500 µmol·L^-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution， while the blank and model groups received an equal volume of saline. Glucose and insulin（INS） levels were measured using enzyme-linked immunosorbent assay（ELISA） kits， the effects on the liver were observed by hematoxylin-eosin（HE） histopathological sections. The mRNA expression levels of glucagon（Glucagon）， insulin（Insa）， and phosphoenolpyruvate carboxykinase 1（PCK1） were detected with real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultsThe optimized extraction conditions were determined as follows：solid-to-liquid ratio of 1∶40（g·mL^-1）， extraction time of 66 min， and extraction temperature of 79 ℃. Under these conditions， the yield of turnip polysaccharides was （10.34±0.96）%. UV analysis indicated that BP-1 contained no proteins or nucleic acids， GC-MS analysis revealed that BP-1 consisted of six monosaccharides（arabinose， rhamnose， ribose， mannose， galactose and glucose）. Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure， FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids， SEM analysis revealed an irregular flaky structure with a flat and smooth surface， AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L^-1. Pharmacodynamic results showed that， compared with the blank group， the model group exhibited significantly increased glucose levels and significantly decreased INS levels（P<0.01）. Compared with the model group， the BP-1-50 group significantly reduced glucose levels and increased INS levels（P<0.05）. Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal， with hepatocytes appearing plump. Real-time PCR results showed that， compared with the blank group， the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1， and significantly downregulated mRNA expression of Insa（P<0.01）. Compared with the model group， the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1， and significantly upregulated mRNA expression of Insa（P<0.01）. ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate， is simple to operate， has low costs， making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides， contains α-glycosidic bonds and uronic acids， exhibits hypoglycemic activity， and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.

As an important setting for the administration of vaccinations, the reasonable setting up and standardized management of vaccination clinics will enhance immunization service quality, public satisfaction, and improve the vaccination rate to protect people′s health. In recent years, various provinces in China are continuously promoting the standardized construction and management of vaccination clinics. However, the level of standardization management remains unbalanced, and the capacity of vaccination services needs to be further improved. This paper reviews the standardized management process of vaccination clinics, summarizes the practice and achievements in various regions, and analyzes the challenges and issues during these processes, to provide reference for improving the standardized management level of vaccination clinics in the future.

Objective To assess the effectiveness of the Regulations on Medical Dispute Prevention and Resolution(hereafter referred to as the Regulations)and to provide evidence-based recommendations for enhancing the legal governance sys-tem of medical dispute management.Methods A cross-sectional study was conducted involving physicians,nurses,techni-cians,clinical department directors,and head nurses.The investigation was conducted through literature review,questionnaire surveys,and expert interviews.Factor analysis and chi-square tests were employed for statistical analysis.Results Significant differences(P＜0.01)were observed among healthcare practitioners in Guangxi concerning their understanding of the Regula-tions,preferences for dispute resolution methods,implementation of informed consent,and risk intervention practices.However,no significant differences emerged in medical quality and safety evaluations or recommendations for surgical accident insurance.Conclusion This study suggests it is a need to refine the legal framework for medical dispute prevention and resolution.It is rec-ommended to strengthen medical personnel's compliance with informed consent obligations and deepen their understanding of rel-evant laws and regulations.Efforts should be intensified to promote third-party mediation mechanisms such as the Medical Dispute Mediation Committee(MedMC)and medical accident insurance coverage.Additionally,pre-dispute risk assessments should be enhanced,and a risk early intervention model integrating artificial intelligence,healthcare practices,and legal regulations should be established.

Objective To establish an electrical field(EF)stimulation model for Schwann cells(SCs),and to provide a basis for exploring the mechanisms of EF stimulation in promoting proliferation,migration and epithelial-to-mesenchymal transition of SCs.Methods A YC-3 bipolar programmable electrical stimulator and an electrotaxis chamber were used to construct an EF stimulation system to stimulate SCs.In the study,SCs were divided into control group(Ctrl)receiving no EF stimulation and EF group stimulated by continuous constant-voltage EF(100 mV/mm,3 h).The effects of EF stimulation on the proliferation and migration of SCs were analyzed using CCK-8 assay,and wound healing assay+Transwell assay,separately;and its effect on SCs adhesion was observed by analyzing the expressions of E-cadherin and N-cadherin using Western Blot.Results The CCK-8 assay results suggested that the absorbance at 450 nm was significantly higher in EF group than in Ctrl group(P<0.05).The results of wound healing assay+Transwell assay revealed that EF group had higher cell migration efficiency than Ctrl group(P<0.05).Western Blot results showed decreased E-cadherin expression and increased N-cadherin expression in EF group as compared with Ctrl group(P<0.05).Conclusion The improved EF stimulation system for SCs is operable.EF stimulation can promote the proliferation and migration of SCs.The decreased E-cadherin expression and increased N-cadherin expression may be related to the occurrence of epithelial-to-mesenchymal transition in SCs after EF stimulation.

As an important setting for the administration of vaccinations, the reasonable setting up and standardized management of vaccination clinics will enhance immunization service quality, public satisfaction, and improve the vaccination rate to protect people′s health. In recent years, various provinces in China are continuously promoting the standardized construction and management of vaccination clinics. However, the level of standardization management remains unbalanced, and the capacity of vaccination services needs to be further improved. This paper reviews the standardized management process of vaccination clinics, summarizes the practice and achievements in various regions, and analyzes the challenges and issues during these processes, to provide reference for improving the standardized management level of vaccination clinics in the future.

Objective To assess the effectiveness of the Regulations on Medical Dispute Prevention and Resolution(hereafter referred to as the Regulations)and to provide evidence-based recommendations for enhancing the legal governance sys-tem of medical dispute management.Methods A cross-sectional study was conducted involving physicians,nurses,techni-cians,clinical department directors,and head nurses.The investigation was conducted through literature review,questionnaire surveys,and expert interviews.Factor analysis and chi-square tests were employed for statistical analysis.Results Significant differences(P＜0.01)were observed among healthcare practitioners in Guangxi concerning their understanding of the Regula-tions,preferences for dispute resolution methods,implementation of informed consent,and risk intervention practices.However,no significant differences emerged in medical quality and safety evaluations or recommendations for surgical accident insurance.Conclusion This study suggests it is a need to refine the legal framework for medical dispute prevention and resolution.It is rec-ommended to strengthen medical personnel's compliance with informed consent obligations and deepen their understanding of rel-evant laws and regulations.Efforts should be intensified to promote third-party mediation mechanisms such as the Medical Dispute Mediation Committee(MedMC)and medical accident insurance coverage.Additionally,pre-dispute risk assessments should be enhanced,and a risk early intervention model integrating artificial intelligence,healthcare practices,and legal regulations should be established.

Objective To establish an electrical field(EF)stimulation model for Schwann cells(SCs),and to provide a basis for exploring the mechanisms of EF stimulation in promoting proliferation,migration and epithelial-to-mesenchymal transition of SCs.Methods A YC-3 bipolar programmable electrical stimulator and an electrotaxis chamber were used to construct an EF stimulation system to stimulate SCs.In the study,SCs were divided into control group(Ctrl)receiving no EF stimulation and EF group stimulated by continuous constant-voltage EF(100 mV/mm,3 h).The effects of EF stimulation on the proliferation and migration of SCs were analyzed using CCK-8 assay,and wound healing assay+Transwell assay,separately;and its effect on SCs adhesion was observed by analyzing the expressions of E-cadherin and N-cadherin using Western Blot.Results The CCK-8 assay results suggested that the absorbance at 450 nm was significantly higher in EF group than in Ctrl group(P<0.05).The results of wound healing assay+Transwell assay revealed that EF group had higher cell migration efficiency than Ctrl group(P<0.05).Western Blot results showed decreased E-cadherin expression and increased N-cadherin expression in EF group as compared with Ctrl group(P<0.05).Conclusion The improved EF stimulation system for SCs is operable.EF stimulation can promote the proliferation and migration of SCs.The decreased E-cadherin expression and increased N-cadherin expression may be related to the occurrence of epithelial-to-mesenchymal transition in SCs after EF stimulation.

Objective To explore whether the environmental pollutant triclosan(TCS)has negative effects on the various biological characteristics of dental pulp stem cells(DPSCs),as well as the distribution and hazards of TCS in rat dental pulp tissue in vivo,which will provide a basis for the clinical application of DPSCs and the safety of TCS.Methods Tooth collection was approved by the Ethics Committee of Tianyou Hospital Affiliated to Wuhan University of Science and Technology.Human DPSCs were extracted,cultured,and identified.Up to 0.08 mmol/L of TCS was added to the in vitro culture medium of DPSCs.The proliferation ability of DPSCs was detected by CCK-8.The migration ability of DPSCs was detected via scratch assay.The differentiation ability of DPSCs was detected by inducing trilineage differenti-ation.The gene or protein expression levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-10(IL-10),inducible nitric oxide synthase(iNOS),and transforming growth factor-β(TGF-β)in DPSCs were detected.The level of reactive oxygen species(ROS)generated by DPSCs was analyzed using fluorescence staining.Changes in mitochondrial membrane potential of DPSCs were detected using a fluorescent probe.The activity of PI3K/Akt/mTOR,p38,and JNK pathways of DPSCs were detected.Animal experiments were approved by the Animal Ethics Committee of Wuhan University of Science and Technology.A rat model of short-term oral exposure to 50 mg/kg/d of TCS for 2 months was established,and the TCS concentration in the liver,brain,and dental pulp tissues of rats was detected through liquid chromatography-mass spectrometry.Results TCS at 0.02 mmol/L,0.04 mmol/L,and 0.08 mmol/L significantly inhibited the proliferation ability of human-derived DPSCs on the 5th and 7th days of contact.TCS at 0.04 mmol/L and 0.08 mmol/L significantly inhibited the migration ability and tri-lineage differentiation ability of DPSCs on the 3rd day of contact.TCS induced the gene or protein expression of proinflammatory factors including TNF-α,IL-1β,IL-6,and iNOS,induced the gene or protein expression of TGF-β,and inhibited the protein expression of anti-inflammatory factor IL-10.On day 1,TCS at 0.04 mmol/L and 0.08 mmol/L induced the production of ROS in DPSCs and reduced the mitochondrial membrane potential of DPSCs.On day 3,TCS at these levels inhibited PI3K/Akt/mTOR pathway activity and enhanced p38 pathway activity of DPSCs,without affecting the pathway activity of JNK.After short-term intragastric exposure of rats to TCS,TCS was detected in liver(430 ng/mL)and brain(41.4 ng/mL)tissues but not in the dental pulp.The TCS concentration was highest in the liver,but no obvious histopathological changes were observed.Conclusion TCS inhibits a variety of biological characteristics of DPSCs and poses a potential risk to the organism.No TCS exists in the dental pulp tissue of rats exposed to TCS for a brief period of time,and the health of the rats is not damaged.

Malocclusion,identified by the World Health Organization(WHO)as one of three major oral diseases,profoundly impacts the dental-maxillofacial functions,facial esthetics,and long-term development of～260 million children in China.Beyond its physical manifestations,malocclusion also significantly influences the psycho-social well-being of these children.Timely intervention in malocclusion can foster an environment conducive to dental-maxillofacial development and substantially decrease the incidence of malocclusion or reduce the severity and complexity of malocclusion in the permanent dentition,by mitigating the negative impact of abnormal environmental influences on the growth.Early orthodontic treatment encompasses accurate identification and treatment of dental and maxillofacial morphological and functional abnormalities during various stages of dental-maxillofacial development,ranging from fetal stages to the early permanent dentition phase.From an economic and societal standpoint,the urgency for effective early orthodontic treatments for malocclusions in childhood cannot be overstated,underlining its profound practical and social importance.This consensus paper discusses the characteristics and the detrimental effects of malocclusion in children,emphasizing critical need for early treatment.It elaborates on corresponding core principles and fundamental approaches in early orthodontics,proposing comprehensive guidance for preventive and interceptive orthodontic treatment,serving as a reference for clinicians engaged in early orthodontic treatment.