ObjectiveTo analyze the pharmacodynamic substance basis of Shangkeling Spray and its potential mechanisms in intervening knee osteoarthritis （KOA） using ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS）， network pharmacology， and molecular docking technology. MethodsUPLC-MS was used to identify the chemical components of Shangkeling Spray. Pharmacokinetic properties were employed to screen potential active ingredients. Network pharmacology methods were utilized to collect potential targets of these ingredients and the pathological gene set of KOA. An "active ingredient-disease" target network was constructed using databases such as STRING. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） functional enrichment analyses were performed using clusterProfiler. Libraries including NumPy were employed to calculate shortest path lengths to identify dominant pharmacodynamic links. Core gene clusters were identified using MCODE， validated through the Gene Expression Omnibus （GEO） database， and molecular docking was performed between key active ingredients and core targets. ResultsA total of 322 and 314 chemical components were identified under positive and negative ion modes， respectively， with 410 components in total after de-duplication， mainly including flavonoids， coumarins， terpenoids， organic acids， and alkaloids. Analysis of the "active ingredient-disease" network identified "development and regeneration"， "cell growth and death"， "immune system"， and "nervous system" as the dominant pharmacodynamic links of Shangkeling Spray in the treatment of KOA. Molecular docking showed that key active ingredients， such as bletillin A， formononetin， morin， oxymatrine， aconitine， gallic acid， curdione， apigenin， naringenin， and oleanolic acid， tightly bound to functional domains of 10 key targets including Jun proteins（JUN）， interleukin-6 （IL-6）， protein kinase B1 （Akt1）， Caspase-3， nuclear transcription factor-κB subunit p65（RELA）， nuclear factor-kappaB1（NF-κB1）， Cyclin D₁， mammalian target of rapamycin（mTOR）， tumor necrosis factor （TNF）， and Fos proto-oncogene protein （FOS）. These interactions synergistically regulated the phosphatidylinositol 3-kinase （PI3K）/Akt/mTOR-related signaling axis and nervous system-related pathways， mediating cartilage repair， reducing inflammation and pain， and improving KOA. ConclusionThis study preliminarily clarifies the pharmacodynamic substance basis of Shangkeling Spray and suggests that its main active ingredients may improve KOA by synergistically regulating the PI3K/Akt/mTOR-related pathways， providing a reference for subsequent exploration of its substance benchmark and mechanism of action.

ObjectiveTo explore the relationship between negative life events and depression severity in adolescent patients with depressive disorder， as well as the chain mediating role of insomnia symptoms and quality of life. Methods374 outpatient patients and hospitalized patients with adolescent depressive disorders were enrolled. The Adolescent Life Event Scale （ASLEC）， the Insomnia Severity Index （ISI）， the World Health Organization Quality of Life Questionnaire Short Form （WHOQOL-BREF）， and the Center for Epidemiology Depression Scale （CES-D） were used to evaluate the negative life event situation， insomnia symptoms， quality of life level and depression severity of the subjects， respectively. In addition， the PROCESS 4.0 macroprogram was used to analyze the chain mediating effect of insomnia symptoms and quality of life between negative life events and depression severity in patients with adolescent depressive disorder. ResultsThe results of correlation analysis showed that there was a significant correlation between negative life events and insomnia symptoms， quality of life， and depression severity （all P<0.05）. In addition， the results of chain mediation showed that negative life events had a significant direct effect on depression severity， with an effect size of 0.12 （P<0.001）. Insomnia symptoms and quality of life played a mediating role in the relationship between negative life events and depression severity in patients with adolescent depressive disorders， with indirect effect sizes of 0.062 （95%CI： 0.040-0.087） and 0.091 （95%CI： 0.059-0.123）， respectively. It could also play a chain mediation role， and the effect size was 0.039 （95%CI： 0.024-0.057）. ConclusionNegative life events experienced by patients with adolescent depressive disorder not only directly affect the severity of depressive symptoms， but may also indirectly exacerbate depression through insomnia symptoms and quality of life.

Objective:To summarize the genotype and clinical phenotype characteristics of children with epilepsy associated with the SCN1B gene encoding the sodium channel β1 subunit. Methods:The genotypes and clinical phenotypes of patients with SCN1B variants among suspected genetic epilepsy cases treated at the Children′s Medical Center of Peking University First Hospital between May 2016 and July 2024 were analyzed. These variants were identified using next-generation sequencing and subsequently validated by Sanger sequencing or quantitative polymerase chain reaction methods. Results:A total of 17 patients were analyzed, including 8 males and 9 females. Ten cases of missense variations (including 2 with the same variations), 4 cases of deletion variations, and 1 case each of nonsense variations, splice site variations, and exons 4-5 deletions were identified. Among them, 6 cases had novel SCN1B variations. The variants in 11 cases were inherited from 1 parent. Eleven types of gene variants have not been reported yet. Onset of epilepsy ranged from 3 months to 5 years and 3 months old (median age: 14 months). Types of seizures included generalized tonic-clonic seizures (GTCS) in 14 cases, focal seizures in 9 cases, myoclonic seizures in 3 cases, atypical absence seizures in 2 cases and epilepsy spasms, tonic seizures and atonic seizures in 1 case each. Eleven cases had diverse seizure types. Fourteen cases (14/17) demonstrated fever sensitivity. Electroencephalography revealed focal discharges in 3 cases, coinciding with focal and generalized discharges in 3 additional cases, and multifocal discharges in 6 cases. Seizures were identified in 4 cases: 1 case of myoclonic seizures, 1 case of GTCS, 1 case of atypical absence seizures, and 1 case exhibiting both myoclonic and tonic seizures. Nine cases (9/17) were diagnosed with genetic epilepsy with febrile seizures plus, 1 case diagnosed with myoclonic epilepsy in infancy and 1 diagnosed with infant epileptic spasms syndrome. There were 2 cases of nonspecific developmental epileptic encephalopathy, while the remaining 4 cases could not be diagnosed with a specific epileptic syndrome. Effective antiseizure medications (ASMs) included valproate in 8 cases, levetiracetam in 5 cases, topiramate in 3 cases, clobazam in 2 cases, clonazepam and vigabatrin in 1 case each. Sodium channel blockers exacerbated seizures in 3 cases, specifically oxcarbazepine in 2 cases and lamotrigine in 1 case. At the last follow-up, seizures were controlled for at least 6 months in 14 patients (14/17), while seizures remained uncontrolled in 3 patients despite trialing 2 or more ASMs. Thirteen patients exhibited normal development, while 4 experienced developmental delays. Conclusions:The heterozygous variants in children with SCN1B gene-related epilepsy include missense, deletion, nonsense, splice site variants, and exon deletions. The correlation between different genetic variants and clinical phenotypes remains unclear. These variants are associated with epilepsy onset from infancy to early childhood, presenting with various seizure types, with GTCS being the most common. Phenotypic manifestations can vary significantly in severity, ranging from benign febrile seizures or febrile seizures plus to developmental epileptic encephalopathy. Valproic acid demonstrates the highest effectiveness rate, while the use of sodium channel blockers may worsen seizures in certain patients, necessitating cautious administration.

Objective:To construct four recombinant hemagglutinin (HA) antigens from seasonal influenza viruses and evaluate their immunogenicity in mouse models.Methods:HA coding sequences from four seasonal influenza virus strains Wisconsin (H1N1), Darwin (H3N2), Austria (B/Victoria lineage, BV) and Phuket (B/Yamagata lineage, BY) were optimized and synthesized, and then used to construct four recombinant plasmids. Recombinant baculoviruses were obtained through transformation and transfection. The expression of recombinant HA antigens was identified by SDS-PAGE and Western blot. The recombinant HA antigens were purified by nickel column affinity chromatography and intramuscularly administered to BALB/c mice after formulation with Al(OH) 3 or AddaVax adjuvant. Humoral immune responses were assessed by indirect ELISA and hemagglutination inhibition test, while cellular immune responses were evaluated by ELISPOT. Microneutralization test was used to detect the titers of serum antibodies in mice. Statistical analysis was performed using t test or non-parametric rank sum test. Results:PCR amplification and agarose gel electrophoresis confirmed the correct construction of the recombinant bacmids. Western blot showed verified the successful expression of the four recombinant antigens (H1-HA, H3N2-HA, BV-HA, and BY-HA). SDS-PAGE results showed that the purity of all four recombinant HA antigens exceeded 95%. After three-dose immunization, the total IgG levels in mice immunized with the recombinant H1N1-HA, H3N2-HA, or BV-HA formulated with AddaVax adjuvant were higher than those in the corresponding groups immunized with the same recombinant antigen alone (all P<0.05). The secretion levels of IFN-γ, IL-2, and IL-4 in the group receiving the mixture of all four recombinant HA antigens formulated with AddaVax adjuvant were higher than those in the group immunized with a commercial quadrivalent split influenza vaccine (all P<0.01). Results of the microneutralization test showed that the antibody titer in the quadrivalent split influenza vaccine group was 1∶225, whereas the titer in the group immunized with the mixture of four recombinant HA antigens formulated with AddaVax adjuvant could reach up to 1∶1 200. Conclusions:In this study, four recombinant seasonal influenza virus HA antigens are successfully expressed and demonstrated good immunogenicity in mice when formulated AddaVax adjuvant.

Objective:To explore the genetic and clinical characteristics of epilepsy related with ATP6V1A gene heterozygous variants.Methods:A case series study was conducted. The clinical data of 10 children of epilepsy associated with ATP6V1A gene variants who were admitted to the Children′s Medical Center, Peking University First Hospital from January 2019 to December 2024 was collected. The characteristics of children′ gene variation, clinical phenotype, auxiliary examination results, treatment and prognosis were analyzed.Results:Among the 10 children, there were 4 boys and 6 girls. All 10 children with ATP6V1A gene variants were de novo heterozygous variants, including 1 case of mosaic variant. A total of 9 different variants were identified and 7 variants have not been reported previously. The age at epilepsy onset was 28 (9, 48) months. Five children experienced their first seizure as a fever induction. The types of epileptic seizures included focal seizures in 6 children, epileptic spasms in 5 children, tonic spasms and atonic seizures in 1 child respectively. Three children had 2 seizure types. Global developmental delays were exhibited in 8 children, 2 of whom manifested autism spectrum disorder phenotypes. Two children showed normal development. Electroencephalography revealed slowed background activity in 5 children. Interictal epileptiform discharges were recorded in 9 cases, including hypsarrhythmia, focal, multifocal or generalized discharges. Clinical seizures were captured in 4 children. Brain magnetic resonance imaging abnormalities were found in 4 children, including frontotemporal cortical dysplasia, prominent sulci, delayed myelination of white matter, dysplasia of the corpus callosum, bilateral ventricular enlargement, and cerebral atrophy. Five children were diagnosed with developmental and epileptic encephalopathy (DEE), and 4 of them were diagnosed with infantile epileptic spasms syndrome. At the last follow-up, the age was 78 (25, 120) months. Seizures were controlled in 6 children, while 4 children had uncontrolled seizures despite treatment with ≥3 anti-seizure medications. Conclusions:All children with ATP6V1A gene related epilepsy harbored de novo heterozygous missense variants, with few showing mosaic variants. Seizure onset age ranged widely from the neonatal period to childhood. The predominant seizure types were focal seizures and epileptic spasms. The phenotypic spectrum may exhibit DEE, while a minority maintain normal development.

OBJECTIVE To explore the risk factors for peritoneal dialysis-associated peritonitis(PDAP)in the type 2 diabetes mellitus patients complicated with renal anemia and construct the risk prediction model so as to provide references for clinical diagnosis and treatment of PDAP.METHODS A total of 142 type 2 diabetes mellitus patients who were complicated with renal anemia and underwent peritoneal dialysis in nephrology department of the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine from Jan.2017 to Dec.2023 were recrui-ted as the research subjects and were divided into the peritonitis group with 68 cases and the non-peritonitis group with 74 cases according to the status of complication with PDAP.The related data were collected,the univariate a-nalysis and binary logistic regression analysis were performed,the risk prediction model was established,and the obtained model was visualized for further analysis.RESULTS The univariate analysis showed that the age and lev-els of glycosylated hemoglobin,triglyceride and fibrinogen were higher in the peritonitis group than in the non-per-itonitis group;the levels of serum potassium,serum iron and total iron binding capacity of the peritonitis group were lower than those of the non-peritonitis group,and there were significant differences(all P＜0.05).The bina-ry logistic regression analysis indicated that glycosylated hemoglobin no less than 7.00％(OR=4.047,95％CI:1.663 to 9.847,P=0.002),no less than 60 years of age(OR=2.181,95％CI:1.039 to 4.578,P=0.039),triglyc-eride greater than 1.47 mmol/L(OR=2.393,95％CI:1.140 to 5.026,P=0.021)and serum iron less than 7.90μmol/L(OR=2.582,95％CI:1.188 to 5.608,P=0.017)were the risk factors for the PDAP in the type 2 diabe-tes mellitus patients complicated with renal anemia.CONCLUSIONS The glycosylated hemoglobin no less than 7.00％,no less than 60 years of age,triglyceride greater than 1.47 mmol/L and serum iron less than 7.90 μmol/L are the risk factors for PDAP in the type 2 diabetes mellitus patients complicated with renal anemia.The risk pre-diction model can be established based on the above factors,and the intervention measures should be actively taken so as to reduce the risk of PDAP and improve the quality of life.

As antibiotic resistance becomes increasingly concerning,antimicrobial peptides,as a new type of antibiotic alternative,have attracted more attention.However,the low enzymatic stability of antimicrobial peptides severely limits their clinical applications.To address this issue,researchers have developed various structural modification strategies,including the introduction of unnatural amino acids,peptide chain cyclization and chemical group modification.This article reviews the basic principles and cases of the above modification strategies analyzes the advantages and disadvantages of different modification strategies and recommends ways these strategies can be optimized.In addition,this article predicts the developments of and potential challenges to strategies for enhancing enzymatic stability of antimicrobial peptides in the hope of providing references for subsequent research and development of antimicrobial peptides.

As antibiotic resistance becomes increasingly concerning,antimicrobial peptides,as a new type of antibiotic alternative,have attracted more attention.However,the low enzymatic stability of antimicrobial peptides severely limits their clinical applications.To address this issue,researchers have developed various structural modification strategies,including the introduction of unnatural amino acids,peptide chain cyclization and chemical group modification.This article reviews the basic principles and cases of the above modification strategies analyzes the advantages and disadvantages of different modification strategies and recommends ways these strategies can be optimized.In addition,this article predicts the developments of and potential challenges to strategies for enhancing enzymatic stability of antimicrobial peptides in the hope of providing references for subsequent research and development of antimicrobial peptides.

OBJECTIVE To explore the risk factors for peritoneal dialysis-associated peritonitis(PDAP)in the type 2 diabetes mellitus patients complicated with renal anemia and construct the risk prediction model so as to provide references for clinical diagnosis and treatment of PDAP.METHODS A total of 142 type 2 diabetes mellitus patients who were complicated with renal anemia and underwent peritoneal dialysis in nephrology department of the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine from Jan.2017 to Dec.2023 were recrui-ted as the research subjects and were divided into the peritonitis group with 68 cases and the non-peritonitis group with 74 cases according to the status of complication with PDAP.The related data were collected,the univariate a-nalysis and binary logistic regression analysis were performed,the risk prediction model was established,and the obtained model was visualized for further analysis.RESULTS The univariate analysis showed that the age and lev-els of glycosylated hemoglobin,triglyceride and fibrinogen were higher in the peritonitis group than in the non-per-itonitis group;the levels of serum potassium,serum iron and total iron binding capacity of the peritonitis group were lower than those of the non-peritonitis group,and there were significant differences(all P＜0.05).The bina-ry logistic regression analysis indicated that glycosylated hemoglobin no less than 7.00％(OR=4.047,95％CI:1.663 to 9.847,P=0.002),no less than 60 years of age(OR=2.181,95％CI:1.039 to 4.578,P=0.039),triglyc-eride greater than 1.47 mmol/L(OR=2.393,95％CI:1.140 to 5.026,P=0.021)and serum iron less than 7.90μmol/L(OR=2.582,95％CI:1.188 to 5.608,P=0.017)were the risk factors for the PDAP in the type 2 diabe-tes mellitus patients complicated with renal anemia.CONCLUSIONS The glycosylated hemoglobin no less than 7.00％,no less than 60 years of age,triglyceride greater than 1.47 mmol/L and serum iron less than 7.90 μmol/L are the risk factors for PDAP in the type 2 diabetes mellitus patients complicated with renal anemia.The risk pre-diction model can be established based on the above factors,and the intervention measures should be actively taken so as to reduce the risk of PDAP and improve the quality of life.

Objective:To construct four recombinant hemagglutinin (HA) antigens from seasonal influenza viruses and evaluate their immunogenicity in mouse models.Methods:HA coding sequences from four seasonal influenza virus strains Wisconsin (H1N1), Darwin (H3N2), Austria (B/Victoria lineage, BV) and Phuket (B/Yamagata lineage, BY) were optimized and synthesized, and then used to construct four recombinant plasmids. Recombinant baculoviruses were obtained through transformation and transfection. The expression of recombinant HA antigens was identified by SDS-PAGE and Western blot. The recombinant HA antigens were purified by nickel column affinity chromatography and intramuscularly administered to BALB/c mice after formulation with Al(OH) 3 or AddaVax adjuvant. Humoral immune responses were assessed by indirect ELISA and hemagglutination inhibition test, while cellular immune responses were evaluated by ELISPOT. Microneutralization test was used to detect the titers of serum antibodies in mice. Statistical analysis was performed using t test or non-parametric rank sum test. Results:PCR amplification and agarose gel electrophoresis confirmed the correct construction of the recombinant bacmids. Western blot showed verified the successful expression of the four recombinant antigens (H1-HA, H3N2-HA, BV-HA, and BY-HA). SDS-PAGE results showed that the purity of all four recombinant HA antigens exceeded 95%. After three-dose immunization, the total IgG levels in mice immunized with the recombinant H1N1-HA, H3N2-HA, or BV-HA formulated with AddaVax adjuvant were higher than those in the corresponding groups immunized with the same recombinant antigen alone (all P<0.05). The secretion levels of IFN-γ, IL-2, and IL-4 in the group receiving the mixture of all four recombinant HA antigens formulated with AddaVax adjuvant were higher than those in the group immunized with a commercial quadrivalent split influenza vaccine (all P<0.01). Results of the microneutralization test showed that the antibody titer in the quadrivalent split influenza vaccine group was 1∶225, whereas the titer in the group immunized with the mixture of four recombinant HA antigens formulated with AddaVax adjuvant could reach up to 1∶1 200. Conclusions:In this study, four recombinant seasonal influenza virus HA antigens are successfully expressed and demonstrated good immunogenicity in mice when formulated AddaVax adjuvant.