Objective:Comprehensive characterization of B-cell phenotypes and spatial distribution in oral lichen planus (OLP) and related oral lichenoid lesions (OLL)(OLP/OLL), with an emphasis on transcriptomic profiling and functional analysis, to uncover the epigenetic mechanisms underlying B cell-mediated immune regulation within the oral mucosal microenvironment.Methods:Single-cell RNA sequencing raw data were sourced from the GSE211630 database, encompassing samples from 2 cases of erosive OLP (EOLP), 3 cases of non-erosive OLP (NEOLP) and 1 healthy control (NORMAL). Following stringent quality control, the data underwent normalization, selection of highly variable genes and batch effect correction. Subsequent analyses included dimensionality reduction and unsupervised clustering to identify distinct cell populations. This study collected pathological specimens from 3 OLP/OLL patients and 3 healthy controls who were treated at the Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from January 2021 to December 2023. Using 10X Genomics Visium HD spatial transcriptomics technology, tissue sections were processed through dewaxing, staining and histological imaging, enabling the reconstruction of nucleic acid structures and the capture of gene expression profiles. Data analysis included quality assessment, gene quantification, normalization, dimensionality reduction and clustering. Furthermore, cell type deconvolution was performed using the robust cell type decomposition algorithm, integrating single-cell transcriptomic data to accurately predict and spatially resolve cell type distributions within the tissue microenvironment.Results:After integrating single-cell data from EOLP, NEOLP and NORMAL, cells were classified into seven major categories: B/plasma cells, endothelial cells, epithelial cells, fibroblasts, myeloid cells, smooth muscle cells and T/natural killer cells. The proportion of B/plasma cells varied significantly among the three groups, accounting for 10.7% (1 693/15 815), 3.8% (833/21 653) and 0.4% (47/11 556) of the total cells respectively. Further clustering analysis of B/plasma cells identified four distinct subpopulations: naive B cells, activated B cells, memory B cells and plasma cells. In the EOLP group, these subpopulations constituted 25.9% (348/1 344), 45.9% (617/1 344), 3.3% (45/1 344) and 24.9% (334/1 344) of the B/plasma cells respectively. In the NEOLP group, they represented 31.6% (195/617), 59.6% (368/617), 0.2% (1/617) and 8.6% (53/617). Howerer, only plasma cells were detected in the NORMAL group. Spatial analysis revealed that B cells were actively involved in the formation of tertiary lymphoid structures (TLS) at various stages in OLP/OLL samples, with a prominent structural organization observed in secondary follicle-like TLS. Within these structures, the expressions of T cells marker gene CD3E and B cells marker gene MS4A1 were significantly elevated. Additionally, in secondary follicle-like TLS, the gene encoding follicular dendritic cell secreted protein, germinal center marker gene B cell lymphoma 6 and the gene for activation induced cytidine deaminase also showed strong expression. In OLP/OLL samples, plasma cell marker gene CD38, immunoglobulin (IGH) G3, IGHG1, IGHM, IGHD, IGHE, imunoglobulin Kappa constant, immunoglobulin alpha 1, immunoglobulin Lambda constant 1 and complement gene C3 all exhibited high levels of expression.Conclusions:Compared to normal mucosa, extensive B-cell infiltration is observed in both OLP and OLL, accompanied by significant differences in B-cell phenotypes and proportions. B cells appear to play a central role in local immune responses, primarily through the formation of TLS. However, the precise functional mechanisms underlying their involvement require further investigation.

BACKGROUND:Total flavonoids from Sophora flavescens have a variety of pharmacological effects,including anti-inflammatory,immunomodulatory,antioxidant,and anti-hepatic injury,but the therapeutic effects and mechanisms in non-alcoholic fatty liver disease are not clear.OBJECTIVE:To reveal the mechanism of total flavonoids from Sophora flavescens in the treatment of non-alcoholic fatty liver disease using bioinformatics,network pharmacology and zebrafish experimental validation.METHODS:A zebrafish model of non-alcoholic fatty liver disease was constructed to observe lipid accumulation,pathomorphologic changes,and expression of inflammatory genes in the liver of zebrafish after treatment with total flavonoids from Sophora flavescens.The active ingredients of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease-related targets were obtained from TCMSP,Swiss Target Prediction,and Bat-man databases.STRING was used to perform protein-protein interaction network analysis,GO functional enrichment and KEGG pathway enrichment analysis.Based on the GSE33814 dataset,the differentially expressed genes of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease intersection targets were screened out.Correlation analysis and receiver operating characteristic curve were performed using R4.3.2 software.Core genes were verified by the validation set GSE89632.RT-qPCR and western blot assays were performed to verify the expression of core pathway-related genes and proteins.RESULTS AND CONCLUSION:(1)Total flavonoids from Sophora flavescens could improve lipid accumulation in the liver of zebrafish with non-alcoholic fatty liver disease,significantly inhibited the elevation of lipid and aminotransferase levels in zebrafish(P<0.05),and regulated the expression of genes related to inflammation and lipid metabolism.(2)A total of 168 common targets were obtained using the network pharmacology,and top 10 core genes,identified by Cytoscape topology analysis,were HSP90AA1,STAT3,PIK3R1,MAPK1,AKT1,RXRA,PIK3CA,EGFR,JAK2,and ESR1.GO and KEGG analysis pathways mainly included insulin resistance,lipids,and atherosclerosis.There were a total of 59 differentially expressed genes after intersection of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease targets.The receiver operating characteristic curve and validation set analyses yielded six core targets that were significantly different between healthy individuals and patients with non-alcoholic fatty liver disease(P<0.01).(3)RT-PCR and western blot results verified that total flavonoids from Sophora flavescens inhibited the activation of the JAK2/STAT3 signaling pathway in zebrafish.To conclude,total flavonoids from Sophora flavescens may alleviate the inflammatory response through the JAK2/STAT3 signaling pathway,thus inhibiting lipid accumulation and improving non-alcoholic fatty liver disease.

Objective:To discuss the clinical efficacy of low-temperature radiofrequency ablation assisted by endoscopy combined with resection and drainage of cervical abscess for the treatment of congenital pyriform sinus fistula （CPSF） in the acute inflammatory period of children. Methods:Clinical data of 30 patients with CPSF in the acute inflammatory period who received low-temperature radiofrequency ablation assisted by endoscopy under laryngoscope, combined with resection and drainage of cervical abscess, from January 2018 to December 2023 were reviewed. After the operation, patients were followed up closely at different stages. All patients underwent color Doppler ultrasound and electronic laryngoscopy, and the results were analyzed. Results:All 30 children successfully completed the surgery without pharyngeal fistula, dysphagia, perifistula, or distal fistula infection, and the incision in the neck healed well. The follow-up survey ranged from 6 months to 2 years, and no recurrences were observed. Conclusion:Low-temperature radiofrequency ablation assisted by endoscopy combined with resection and drainage of cervical abscess is a promising method for treating CPSF in the acute inflammatory period. It is less traumatic, simple, safe, has a significant curative effect, and a low recurrence rate. This approach can be used as a supplementary operation for CPSF in children and provides a new way for clinical treatment.
Humans ; Pyriform Sinus/abnormalities* ; Abscess/surgery* ; Drainage ; Fistula/congenital* ; Female ; Male ; Child ; Radiofrequency Ablation ; Treatment Outcome ; Postoperative Period ; Endoscopy ; Laryngoscopy ; Inflammation ; Child, Preschool

Objective:Comprehensive characterization of B-cell phenotypes and spatial distribution in oral lichen planus (OLP) and related oral lichenoid lesions (OLL)(OLP/OLL), with an emphasis on transcriptomic profiling and functional analysis, to uncover the epigenetic mechanisms underlying B cell-mediated immune regulation within the oral mucosal microenvironment.Methods:Single-cell RNA sequencing raw data were sourced from the GSE211630 database, encompassing samples from 2 cases of erosive OLP (EOLP), 3 cases of non-erosive OLP (NEOLP) and 1 healthy control (NORMAL). Following stringent quality control, the data underwent normalization, selection of highly variable genes and batch effect correction. Subsequent analyses included dimensionality reduction and unsupervised clustering to identify distinct cell populations. This study collected pathological specimens from 3 OLP/OLL patients and 3 healthy controls who were treated at the Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine from January 2021 to December 2023. Using 10X Genomics Visium HD spatial transcriptomics technology, tissue sections were processed through dewaxing, staining and histological imaging, enabling the reconstruction of nucleic acid structures and the capture of gene expression profiles. Data analysis included quality assessment, gene quantification, normalization, dimensionality reduction and clustering. Furthermore, cell type deconvolution was performed using the robust cell type decomposition algorithm, integrating single-cell transcriptomic data to accurately predict and spatially resolve cell type distributions within the tissue microenvironment.Results:After integrating single-cell data from EOLP, NEOLP and NORMAL, cells were classified into seven major categories: B/plasma cells, endothelial cells, epithelial cells, fibroblasts, myeloid cells, smooth muscle cells and T/natural killer cells. The proportion of B/plasma cells varied significantly among the three groups, accounting for 10.7% (1 693/15 815), 3.8% (833/21 653) and 0.4% (47/11 556) of the total cells respectively. Further clustering analysis of B/plasma cells identified four distinct subpopulations: naive B cells, activated B cells, memory B cells and plasma cells. In the EOLP group, these subpopulations constituted 25.9% (348/1 344), 45.9% (617/1 344), 3.3% (45/1 344) and 24.9% (334/1 344) of the B/plasma cells respectively. In the NEOLP group, they represented 31.6% (195/617), 59.6% (368/617), 0.2% (1/617) and 8.6% (53/617). Howerer, only plasma cells were detected in the NORMAL group. Spatial analysis revealed that B cells were actively involved in the formation of tertiary lymphoid structures (TLS) at various stages in OLP/OLL samples, with a prominent structural organization observed in secondary follicle-like TLS. Within these structures, the expressions of T cells marker gene CD3E and B cells marker gene MS4A1 were significantly elevated. Additionally, in secondary follicle-like TLS, the gene encoding follicular dendritic cell secreted protein, germinal center marker gene B cell lymphoma 6 and the gene for activation induced cytidine deaminase also showed strong expression. In OLP/OLL samples, plasma cell marker gene CD38, immunoglobulin (IGH) G3, IGHG1, IGHM, IGHD, IGHE, imunoglobulin Kappa constant, immunoglobulin alpha 1, immunoglobulin Lambda constant 1 and complement gene C3 all exhibited high levels of expression.Conclusions:Compared to normal mucosa, extensive B-cell infiltration is observed in both OLP and OLL, accompanied by significant differences in B-cell phenotypes and proportions. B cells appear to play a central role in local immune responses, primarily through the formation of TLS. However, the precise functional mechanisms underlying their involvement require further investigation.

Objective:To investigate the efficacy of antiviral therapy and influencing factors of hepatitis B surface antigen(HBsAg) negative conversion for hepatitis B e antigen(HBeAg)-positive chronic hepatitis B(CHB) in children.Methods:The clinical data of 38 children with CHB who received antiviral treatment in Children's Hospital Affiliated to Xi'an Jiaotong University from January 2019 to August 2024 were collected.All patients were treated with interferon alpha monotherapy or combined with nucleoside analogues for 48 weeks.The patients were divided into HBsAg negative group and HBsAg non-negative group according to the therapeutic results at 48 weeks.Multivariate Logistic regression were used to identify influencing factors of HBsAg negative conversion at 48 weeks.The receiver operator characteristic（ROC）curve was used to analyze the predictive value of each factor to HBsAg negative conversion.Results:The alanine aminotransferase normalization rate，hepatitis B virus DNA negative rate，HBeAg negative rate and HBsAg negative rate were 76.3%，94.7%，39.5% and 47.4%，respectively at 48 weeks.There were 18 cases in HBsAg negative group and 20 cases in HBsAg non-negative group.There were statistical significant differences in age and HBsAg decline level at 12 and 24 weeks of antiviral treatment between HBsAg negative group and HBsAg non-negative group（ P<0.05）.Multivariate Logistic regression analysis showed that age and HBsAg decline level at 12 and 24 weeks of antiviral treatment were independent predictors of HBsAg negative conversion at 48 weeks（ OR=0.664，95% CI 0.473-0.932， P=0.018； OR=8.719，95% CI 1.920-39.604， P=0.005； OR=6.182，95% CI 2.083-18.347， P=0.001）.The area under the curve of age and HBsAg decline level at 12 and 24 weeks were 0.737（95% CI 0.576-0.899， P=0.012），0.847（95% CI 0.725-0.969， P<0.001）and 0.939（95% CI 0.811-0.991， P<0.001），respectively.When the age was less than 4.625 years，the sensitivity，specificity，positive predictive value and negative predictive value of HBsAg negative conversion at 48 weeks were 83.3%，65.0%，68.2% and 81.3%，respectively.A decrease in HBsAg level of >1.07 lg IU/mL at 12 weeks of treatment had a sensitivity，specificity，positive predictive value，and negative predictive value of 72.2%，90.0%，86.7%，and 78.3%，respectively，for predicting HBsAg seroclearance at 48 weeks.A reduction in HBsAg of >1.92 lg IU/mL at 24 weeks of treatment showed a sensitivity，specificity，positive predictive value，and negative predictive value of 83.3%，90.0%，88.2%，and 85.7%，respectively，in predicting HBsAg seroclearance at 48 weeks. Conclusion:The children with CHB have a higher rate of HBsAg negative conversion after antiviral therapy at 48 weeks.Age and HBsAg decline level at 12 and 24 weeks of antiviral treatment can serve as early predictors for HBsAg negative conversion in children with CHB.

BACKGROUND:Total flavonoids from Sophora flavescens have a variety of pharmacological effects,including anti-inflammatory,immunomodulatory,antioxidant,and anti-hepatic injury,but the therapeutic effects and mechanisms in non-alcoholic fatty liver disease are not clear.OBJECTIVE:To reveal the mechanism of total flavonoids from Sophora flavescens in the treatment of non-alcoholic fatty liver disease using bioinformatics,network pharmacology and zebrafish experimental validation.METHODS:A zebrafish model of non-alcoholic fatty liver disease was constructed to observe lipid accumulation,pathomorphologic changes,and expression of inflammatory genes in the liver of zebrafish after treatment with total flavonoids from Sophora flavescens.The active ingredients of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease-related targets were obtained from TCMSP,Swiss Target Prediction,and Bat-man databases.STRING was used to perform protein-protein interaction network analysis,GO functional enrichment and KEGG pathway enrichment analysis.Based on the GSE33814 dataset,the differentially expressed genes of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease intersection targets were screened out.Correlation analysis and receiver operating characteristic curve were performed using R4.3.2 software.Core genes were verified by the validation set GSE89632.RT-qPCR and western blot assays were performed to verify the expression of core pathway-related genes and proteins.RESULTS AND CONCLUSION:(1)Total flavonoids from Sophora flavescens could improve lipid accumulation in the liver of zebrafish with non-alcoholic fatty liver disease,significantly inhibited the elevation of lipid and aminotransferase levels in zebrafish(P<0.05),and regulated the expression of genes related to inflammation and lipid metabolism.(2)A total of 168 common targets were obtained using the network pharmacology,and top 10 core genes,identified by Cytoscape topology analysis,were HSP90AA1,STAT3,PIK3R1,MAPK1,AKT1,RXRA,PIK3CA,EGFR,JAK2,and ESR1.GO and KEGG analysis pathways mainly included insulin resistance,lipids,and atherosclerosis.There were a total of 59 differentially expressed genes after intersection of total flavonoids from Sophora flavescens and non-alcoholic fatty liver disease targets.The receiver operating characteristic curve and validation set analyses yielded six core targets that were significantly different between healthy individuals and patients with non-alcoholic fatty liver disease(P<0.01).(3)RT-PCR and western blot results verified that total flavonoids from Sophora flavescens inhibited the activation of the JAK2/STAT3 signaling pathway in zebrafish.To conclude,total flavonoids from Sophora flavescens may alleviate the inflammatory response through the JAK2/STAT3 signaling pathway,thus inhibiting lipid accumulation and improving non-alcoholic fatty liver disease.

Objective:To investigate the efficacy of antiviral therapy and influencing factors of hepatitis B surface antigen(HBsAg) negative conversion for hepatitis B e antigen(HBeAg)-positive chronic hepatitis B(CHB) in children.Methods:The clinical data of 38 children with CHB who received antiviral treatment in Children's Hospital Affiliated to Xi'an Jiaotong University from January 2019 to August 2024 were collected.All patients were treated with interferon alpha monotherapy or combined with nucleoside analogues for 48 weeks.The patients were divided into HBsAg negative group and HBsAg non-negative group according to the therapeutic results at 48 weeks.Multivariate Logistic regression were used to identify influencing factors of HBsAg negative conversion at 48 weeks.The receiver operator characteristic（ROC）curve was used to analyze the predictive value of each factor to HBsAg negative conversion.Results:The alanine aminotransferase normalization rate，hepatitis B virus DNA negative rate，HBeAg negative rate and HBsAg negative rate were 76.3%，94.7%，39.5% and 47.4%，respectively at 48 weeks.There were 18 cases in HBsAg negative group and 20 cases in HBsAg non-negative group.There were statistical significant differences in age and HBsAg decline level at 12 and 24 weeks of antiviral treatment between HBsAg negative group and HBsAg non-negative group（ P<0.05）.Multivariate Logistic regression analysis showed that age and HBsAg decline level at 12 and 24 weeks of antiviral treatment were independent predictors of HBsAg negative conversion at 48 weeks（ OR=0.664，95% CI 0.473-0.932， P=0.018； OR=8.719，95% CI 1.920-39.604， P=0.005； OR=6.182，95% CI 2.083-18.347， P=0.001）.The area under the curve of age and HBsAg decline level at 12 and 24 weeks were 0.737（95% CI 0.576-0.899， P=0.012），0.847（95% CI 0.725-0.969， P<0.001）and 0.939（95% CI 0.811-0.991， P<0.001），respectively.When the age was less than 4.625 years，the sensitivity，specificity，positive predictive value and negative predictive value of HBsAg negative conversion at 48 weeks were 83.3%，65.0%，68.2% and 81.3%，respectively.A decrease in HBsAg level of >1.07 lg IU/mL at 12 weeks of treatment had a sensitivity，specificity，positive predictive value，and negative predictive value of 72.2%，90.0%，86.7%，and 78.3%，respectively，for predicting HBsAg seroclearance at 48 weeks.A reduction in HBsAg of >1.92 lg IU/mL at 24 weeks of treatment showed a sensitivity，specificity，positive predictive value，and negative predictive value of 83.3%，90.0%，88.2%，and 85.7%，respectively，in predicting HBsAg seroclearance at 48 weeks. Conclusion:The children with CHB have a higher rate of HBsAg negative conversion after antiviral therapy at 48 weeks.Age and HBsAg decline level at 12 and 24 weeks of antiviral treatment can serve as early predictors for HBsAg negative conversion in children with CHB.

Disinfection by-products (DBPs) are compounds generated by chemical reactions between disinfectants and chemical substances in the water during the process of drinking water treatment. More than 700 kinds of DBPs have been identified, the most abundant components are trihalomethanes and haloacetic acids. The safety of DBPs has attracted widespread attention. At present, studies have confirmed that DBPs have cytotoxicity, genotoxicity, carcinogenicity and developmental toxicity. In recent years, the damage of DBPs to reproductive function has been an increasing concern. This article reviewed the research progress on the effects of DBPs exposure on adult reproductive function based on toxicological and epidemiological studies, and discussed the future research direction.

Repetitive transcranial magnetic stimulation (rTMS) has become an essential method in psychiatric disorders. However, many problems occurred in clinical application. This article interpreted the Association Standard T/CMEAS 011-2023'Operating Specifications for Repetitive Transcranial Magnetic Stimulation in Clinical Applications on Psychiatric Disorders′ released by the Chinese Medicine Education Association. The main content included a range of applications, normative references, terms and definitions, site specifications, equipment specifications, ability specifications of rTMS operators and rTMS process specifications.This article provided suggestions for clinical applications of rTMS on psychiatric disorders.

Disinfection by-products (DBPs) are compounds generated by chemical reactions between disinfectants and chemical substances in the water during the process of drinking water treatment. More than 700 kinds of DBPs have been identified, the most abundant components are trihalomethanes and haloacetic acids. The safety of DBPs has attracted widespread attention. At present, studies have confirmed that DBPs have cytotoxicity, genotoxicity, carcinogenicity and developmental toxicity. In recent years, the damage of DBPs to reproductive function has been an increasing concern. This article reviewed the research progress on the effects of DBPs exposure on adult reproductive function based on toxicological and epidemiological studies, and discussed the future research direction.