Objective:To explore the genotype and the clinical phenotype of SCN2A-related developmental delay in children. Methods:A case series study was adopted. Collect clinical data from 10 cases of children with SCN2A gene variants diagnosed with global developmental delay/intellectual disability who were admitted to the Children′s Hospital between July 2019 and March 2023. Summarize the clinical phenotype and genotype based on clinical data such as general information, clinical manifestations, imaging examinations, laboratory tests, genetic testing results, and comprehensive pediatric neuropsychological development assessment. Results:A total of 10 patients were recruited, including 7 males and 3 females, with an age range of 27 days to 5 years and 9 months. 9 patients underwent children′s neuropsychological and behavioral assessments, and the results were consistent with global developmental delay, including 2 mild cases, 4 moderate cases, and 3 severe cases. 3 cases had autism spectrum disorder, and 2 cases had epilepsy. 6 patients underwent complete head MRI examination, and 4 of them showed abnormalities, including delayed myelination, widening of the local extra brain space in the frontal lobe, and abnormal frontal lobe morphology. All 10 cases had point variants. Among them, 9 cases are de novo and 1 case is maternal inheritance. Out of 10 cases, there were 5 cases with copy number variations, but all of them were of unknown significance. Among the 10 variants, 8 have been reported and 2 have not been reported, namely c.4145A>T(p.N1382I) and c.4937T>A(p.I1646N). In this study, 4 out of 10 patients with SCN2A variants had variation sites located in the S4 segment of domain which constitute Nav1.2, the sodium ion channel encoded by SCN2A. The developmental quotient level was lower when the variation sites were located in the S4 segment of domain, and the difference was statistically significant ( t=-3.101, P=0.017), indicating that the severity of developmental delay may be related to the localization of amino acids corresponding to variant sites within the protein domain. Conclusion:SCN2A mutations are strongly associated with diverse neurodevelopmental disorders. In this study, the phenotypic spectrum of SCN2A variants encompassed epilepsy, global developmental delay, and autism spectrum disorder. Affected individuals exhibited early-onset developmental delays, predominantly moderate to severe in severity. Voltage-sensing domain dysfunction in sodium channels may constitute a critical pathomechanism underlying neurodevelopmental impairments. Further electrophysiological characterization and molecular mechanistic studies are warranted todelineate the genotype-phenotype correlations between specific variant loci and clinical severity.

Objective To employ three dimensional high-resolution vessel wall magnetic resonance imaging(3D hr-VW-MRI)for analyzing the imaging characteristics of posterior circulation non-stenotic intracranial atherosclerotic plaque and to discuss its diagnostic value in identifying culprit plaques.Methods Ninety-three patients(age[62.94±9.70]years old,67 males,26 females)with non-stenotic atherosclerosis in our hospital from Jan.2019 to Jan.2021 were retrospectively recruited.The imaging features of plaques,including luminal area,maximum wall thickness and minimum wall thickness at the most stenotic site,stenosis rate,plaque burden,remodeling index,eccentricity index,enhancement ratio at the most stenotic site,and intraplaque hemorrhage,were measured based on T1-weighted imaging(T1WI)and contrast-enhanced T1WI.The culprit plaque was defined as a lesion arising from the responsible vascular supply area to a fresh infarction on the diffusion weighted imaging(DWI)and T2 fluid attenuated inversion recovery(T2-FLAIR)images with accompanying ischemic stroke/transient ischemic attack(TIA).A plaque was considered to be a nonculprit plaque when it occurred in patients with presumed ischemic stroke/TIA,but without an infarct on DWI and T2-FLAIR.Results Sixty-one culprit plaques and 32 non-culprit plaques were analyzed.The proportions of patients with hyperlipidemia,National Institutes of Health stroke scale(NIHSS)score,narrowest plaque enhancement rate,and incidence of intraplaque hemorrhage in the culprit plaque group were significantly higher than those in the non-culprit plaque group(all P＜0.05).Multivariate logistic regression analyses showed that NIHSS score(odds ratio[OR]=1.799,95％confidence interval[CI]1.303-2.484,P＜0.001),enhancement ratio(OR=1.076,95％CI 1.027-1.128,P=0.002)and intraplaque hemorrhage(OR=30.708,95％CI 2.563-367.925,P=0.007)were associated with plaque type.Conclusion NIHSS score,enhancement ratio at the most stenotic site,and intraplaque hemorrhage are independent risk factors for culprit plaques in patients with posterior circulation non-stenotic intracranial atherosclerotic disease.These indicators may help identify such culprit plaques and could be used to screen individuals with plaques having these characteristics,thereby providing a basis for early preventive interventions.

Objective To investigate the current status of home medicine cabinet management and unmet needs for home-based pharmaceutical services among elderly hypertensive patients in Hubei Province,and to provide evidence for optimizing home pharmaceutical care models.Methods From July 2022 to March 2023,310 elderly hypertension patients across Hubei Province were enrolled.Pharmacists conducted home visits to administer structured questionnaires capturing demographics and blood pressure control rates,perform medication categorization and quality assessments of home medicine cabinets and document cabinet management issues in real-time.Results Among 290 valid questionnaires,binary logistic regression revealed better blood pressure control in non-working and non-smoking individuals.Medicine cabinet audits identified three critical issues:failure to separate Chinese and chemical medicines,duplicate medications with similar ingredients and missing original packaging.Post-intervention,all cabinets demonstrated proper medication categorization and storage compliance.Conclusions Significant medication safety hazards exist in elderly hypertensive patients'home medicine cabinets.Pharmacist-led home interventions effectively cleared substandard medications and improved medication safety.Future home pharmaceutical services should focus on optimizing cabinet management protocols,enhancing medication safety awareness and personalizing pharmaceutical care to ensure precise and effective home-based medication management.

The novel coronavirus(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2),classified as a single-stranded RNA virus,replicates and transcribes its genome through the action of an RNA-dependent RNA polymerase,which is itself comprised of numerous non-structural pro-teins(non-structural proteins,Nsps).The present study delineates the development of a detection system founded on a bicistronic reporter plasmid in conjunction with an array of Nsp plasmids,ai-ming to investigate the influence of SARS-CoV-2 Nsps on the virus's replication and expression profiles.Specifically,a bicistronic reporter gene plasmid along with twelve distinct Nsp plasmids(encompassing Nsp3C-Flag,Nsp4,Nsp6-Nsp10,Nsp12-Nsp16)were meticulously constructed via molecular cloning techniques.The successful expression of these Nsps was subsequently confirmed through Western blot analysis.Initially,the activity of the RNA-dependent RNA polymerase was assessed by co-transfection of the reporter plasmid with Nsp12,in the presence of its auxiliary fac-tors Nsp7 and Nsp8,with careful regulation of the co-transfection ratio,culturing temperature,and the timing of activity determination for the triad of Nsp plasmids.The normalized NLuc fluorescein value,in reference to the FLuc fluorescein value of the housekeeping gene,served as a metric for determining the polymerase activity.Building upon this foundation,the co-transfection concentra-tions of Nsp9-16 were fine-tuned,followed by the incremental addition of varying doses of Nsp3C,Nsp4,and Nsp6,to further elucidate the activity of RNA-dependent RNA polymerase(RdRp).The findings indicated that upon transfection with varying ratios of Nsp7,Nsp8,and Nsp12 at a propor-tion of 1∶8∶24,the polymerase activity was markedly elevated compared to the control group,with a statistical significance level(P＜0.001).Furthermore,in the absence of Nsp3,Nsp4,and Nsp6,the inclusion of Nsp10-16 substantially augmented the activity of the RdRp,particularly in scenarios where Nsp9 was not introduced,achieving statistical significance(P＜0.001).In the pres-ence of Nsp3 and Nsp4,the RdRp activity was augmented further upon the addition of Nsp9,reac-hing a level of significance(P＜0.05).The data imply that Nsp9 is capable of enhancing the RdRp activity of SARS-CoV-2 exclusively in the context of Nsp3 and Nsp4 coexistence,suggesting that the stimulatory influence of Nsp9 on viral replication may be contingent upon the formation of double-membrane vesicles.

Objective:To explore the potential value of dynamic changes of inflammatory factors in evaluating the efficacy of platinum regimen in the first-line treatment of advanced lung adenocarcinoma(LUAD).Methods:A total of 121 patients with advanced LUAD without common target mutations who were admitted to Dongtai People's Hospital to receive first-line treatment with platinum regimen from January 2021 to January 2023 were selected,and relevant clinical data such as general information,hematuria routine,angiogenic factors,serum tumor markers,inflammatory factors,immunoglobulin and T lymphocyte indexes were collected and statisti-cally analyzed.Patients in complete response(CR)group and partial response(PR)group were classified as chemotherapy sensitive group;stable(SD)group and progressive(PD)group were classified as chemotherapy insensitive group according to response evaluation criteria in solid tumors(RECIST).Through univariate and multi-factor Logistics regression analysis,the influencing factors of first-line treatment efficacy of platinum regimen in patients with advanced LUAD were analyzed,and the regression equation y=1-1/(1+e-z)prediction model was established and verified.Receiver operating characteristic(ROC)curve was used to analyze the potential value of dynamic changes of inflammatory factors in evaluating the efficacy of platinum-based first-line treatment for advanced LUAD.Results:①Univariate and multivariate Logistic regression analysis showed that CEA,TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP were still the influencing factors for the efficacy of first-line chemotherapy of platinum regimen in patients with advanced LUAD(P<0.05).②Inflammatory factors TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP were included in multivariate Logistic regression analysis.After correcting confounders(Model 5),high levels of TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP were positively correlated with insensitivity after first-line chemotherapy of platinum regimen in advanced LUAD patients(P<0.05).Restrictive cubic spline model analysis showed that the dynamic changes of inflammatory factors and the insensitivity of patients with advanced LUAD after first-line chemotherapy all had a nonlinear dose-response relationship.With the increases of TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP levels,patients with advanced LUAD have a greatly increased risk of desensitization after first-line chemotherapy.③Stratified interaction test analysis showed that tumor stage,differentiation degree and lymph node metastasis before and after correction factors were significantly correlated with expressions of inflammatory factors(all P<0.05,all P interaction<0.05).④Prediction model based on the influence factors of inflammatory factors was well distinguished and accurate,the area under ROC curve(AUC)before and after internal validation for detecting chemotherapy sensitivity in patients with advanced LUAD treated with platinum regimen first-line therapy was 0.87(95%CI:0.81～0.93)and 0.88(95%CI:0.82～0.95),respectively,and the sensitivity were 89.69%and 89.75%,respectively,the specificity were 91.77%and 91.85%,respectively.Conclusion:Dynamic changes of inflammatory factors is an important factor affecting the chemotherapy efficacy of patients with advanced LUAD treated with platinum-containing regimen.With the significant decrease of inflammatory factors,the sensitivity of patients to chemotherapy is also significantly increased.

The novel coronavirus(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2),classified as a single-stranded RNA virus,replicates and transcribes its genome through the action of an RNA-dependent RNA polymerase,which is itself comprised of numerous non-structural pro-teins(non-structural proteins,Nsps).The present study delineates the development of a detection system founded on a bicistronic reporter plasmid in conjunction with an array of Nsp plasmids,ai-ming to investigate the influence of SARS-CoV-2 Nsps on the virus's replication and expression profiles.Specifically,a bicistronic reporter gene plasmid along with twelve distinct Nsp plasmids(encompassing Nsp3C-Flag,Nsp4,Nsp6-Nsp10,Nsp12-Nsp16)were meticulously constructed via molecular cloning techniques.The successful expression of these Nsps was subsequently confirmed through Western blot analysis.Initially,the activity of the RNA-dependent RNA polymerase was assessed by co-transfection of the reporter plasmid with Nsp12,in the presence of its auxiliary fac-tors Nsp7 and Nsp8,with careful regulation of the co-transfection ratio,culturing temperature,and the timing of activity determination for the triad of Nsp plasmids.The normalized NLuc fluorescein value,in reference to the FLuc fluorescein value of the housekeeping gene,served as a metric for determining the polymerase activity.Building upon this foundation,the co-transfection concentra-tions of Nsp9-16 were fine-tuned,followed by the incremental addition of varying doses of Nsp3C,Nsp4,and Nsp6,to further elucidate the activity of RNA-dependent RNA polymerase(RdRp).The findings indicated that upon transfection with varying ratios of Nsp7,Nsp8,and Nsp12 at a propor-tion of 1∶8∶24,the polymerase activity was markedly elevated compared to the control group,with a statistical significance level(P＜0.001).Furthermore,in the absence of Nsp3,Nsp4,and Nsp6,the inclusion of Nsp10-16 substantially augmented the activity of the RdRp,particularly in scenarios where Nsp9 was not introduced,achieving statistical significance(P＜0.001).In the pres-ence of Nsp3 and Nsp4,the RdRp activity was augmented further upon the addition of Nsp9,reac-hing a level of significance(P＜0.05).The data imply that Nsp9 is capable of enhancing the RdRp activity of SARS-CoV-2 exclusively in the context of Nsp3 and Nsp4 coexistence,suggesting that the stimulatory influence of Nsp9 on viral replication may be contingent upon the formation of double-membrane vesicles.

Objective:To explore the genotype and the clinical phenotype of SCN2A-related developmental delay in children. Methods:A case series study was adopted. Collect clinical data from 10 cases of children with SCN2A gene variants diagnosed with global developmental delay/intellectual disability who were admitted to the Children′s Hospital between July 2019 and March 2023. Summarize the clinical phenotype and genotype based on clinical data such as general information, clinical manifestations, imaging examinations, laboratory tests, genetic testing results, and comprehensive pediatric neuropsychological development assessment. Results:A total of 10 patients were recruited, including 7 males and 3 females, with an age range of 27 days to 5 years and 9 months. 9 patients underwent children′s neuropsychological and behavioral assessments, and the results were consistent with global developmental delay, including 2 mild cases, 4 moderate cases, and 3 severe cases. 3 cases had autism spectrum disorder, and 2 cases had epilepsy. 6 patients underwent complete head MRI examination, and 4 of them showed abnormalities, including delayed myelination, widening of the local extra brain space in the frontal lobe, and abnormal frontal lobe morphology. All 10 cases had point variants. Among them, 9 cases are de novo and 1 case is maternal inheritance. Out of 10 cases, there were 5 cases with copy number variations, but all of them were of unknown significance. Among the 10 variants, 8 have been reported and 2 have not been reported, namely c.4145A>T(p.N1382I) and c.4937T>A(p.I1646N). In this study, 4 out of 10 patients with SCN2A variants had variation sites located in the S4 segment of domain which constitute Nav1.2, the sodium ion channel encoded by SCN2A. The developmental quotient level was lower when the variation sites were located in the S4 segment of domain, and the difference was statistically significant ( t=-3.101, P=0.017), indicating that the severity of developmental delay may be related to the localization of amino acids corresponding to variant sites within the protein domain. Conclusion:SCN2A mutations are strongly associated with diverse neurodevelopmental disorders. In this study, the phenotypic spectrum of SCN2A variants encompassed epilepsy, global developmental delay, and autism spectrum disorder. Affected individuals exhibited early-onset developmental delays, predominantly moderate to severe in severity. Voltage-sensing domain dysfunction in sodium channels may constitute a critical pathomechanism underlying neurodevelopmental impairments. Further electrophysiological characterization and molecular mechanistic studies are warranted todelineate the genotype-phenotype correlations between specific variant loci and clinical severity.

Objective To investigate the current status of home medicine cabinet management and unmet needs for home-based pharmaceutical services among elderly hypertensive patients in Hubei Province,and to provide evidence for optimizing home pharmaceutical care models.Methods From July 2022 to March 2023,310 elderly hypertension patients across Hubei Province were enrolled.Pharmacists conducted home visits to administer structured questionnaires capturing demographics and blood pressure control rates,perform medication categorization and quality assessments of home medicine cabinets and document cabinet management issues in real-time.Results Among 290 valid questionnaires,binary logistic regression revealed better blood pressure control in non-working and non-smoking individuals.Medicine cabinet audits identified three critical issues:failure to separate Chinese and chemical medicines,duplicate medications with similar ingredients and missing original packaging.Post-intervention,all cabinets demonstrated proper medication categorization and storage compliance.Conclusions Significant medication safety hazards exist in elderly hypertensive patients'home medicine cabinets.Pharmacist-led home interventions effectively cleared substandard medications and improved medication safety.Future home pharmaceutical services should focus on optimizing cabinet management protocols,enhancing medication safety awareness and personalizing pharmaceutical care to ensure precise and effective home-based medication management.

Objective:To explore the potential value of dynamic changes of inflammatory factors in evaluating the efficacy of platinum regimen in the first-line treatment of advanced lung adenocarcinoma(LUAD).Methods:A total of 121 patients with advanced LUAD without common target mutations who were admitted to Dongtai People's Hospital to receive first-line treatment with platinum regimen from January 2021 to January 2023 were selected,and relevant clinical data such as general information,hematuria routine,angiogenic factors,serum tumor markers,inflammatory factors,immunoglobulin and T lymphocyte indexes were collected and statisti-cally analyzed.Patients in complete response(CR)group and partial response(PR)group were classified as chemotherapy sensitive group;stable(SD)group and progressive(PD)group were classified as chemotherapy insensitive group according to response evaluation criteria in solid tumors(RECIST).Through univariate and multi-factor Logistics regression analysis,the influencing factors of first-line treatment efficacy of platinum regimen in patients with advanced LUAD were analyzed,and the regression equation y=1-1/(1+e-z)prediction model was established and verified.Receiver operating characteristic(ROC)curve was used to analyze the potential value of dynamic changes of inflammatory factors in evaluating the efficacy of platinum-based first-line treatment for advanced LUAD.Results:①Univariate and multivariate Logistic regression analysis showed that CEA,TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP were still the influencing factors for the efficacy of first-line chemotherapy of platinum regimen in patients with advanced LUAD(P<0.05).②Inflammatory factors TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP were included in multivariate Logistic regression analysis.After correcting confounders(Model 5),high levels of TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP were positively correlated with insensitivity after first-line chemotherapy of platinum regimen in advanced LUAD patients(P<0.05).Restrictive cubic spline model analysis showed that the dynamic changes of inflammatory factors and the insensitivity of patients with advanced LUAD after first-line chemotherapy all had a nonlinear dose-response relationship.With the increases of TNF-α,IL-6,IL-8,IL-18,IL-1β and hs-CRP levels,patients with advanced LUAD have a greatly increased risk of desensitization after first-line chemotherapy.③Stratified interaction test analysis showed that tumor stage,differentiation degree and lymph node metastasis before and after correction factors were significantly correlated with expressions of inflammatory factors(all P<0.05,all P interaction<0.05).④Prediction model based on the influence factors of inflammatory factors was well distinguished and accurate,the area under ROC curve(AUC)before and after internal validation for detecting chemotherapy sensitivity in patients with advanced LUAD treated with platinum regimen first-line therapy was 0.87(95%CI:0.81～0.93)and 0.88(95%CI:0.82～0.95),respectively,and the sensitivity were 89.69%and 89.75%,respectively,the specificity were 91.77%and 91.85%,respectively.Conclusion:Dynamic changes of inflammatory factors is an important factor affecting the chemotherapy efficacy of patients with advanced LUAD treated with platinum-containing regimen.With the significant decrease of inflammatory factors,the sensitivity of patients to chemotherapy is also significantly increased.

Objective To investigate the mechanism of Dayuan Decoction in the treatment of Helicobacter pylori(Hp)-positive perioral acne based on network pharmacology and animal experiments.Methods The effective active components and their corresponding targets of Dayuan Decoction were obtained from TCMSP.Targets related to Hp and perioral acne were retrieved from GeneCards,OMIM and TTD databases,and the intersection of drug targets and disease targets was considered as the target of Dayuan Decoction in the treatment of Hp-positive perioral acne.A protein-protein interaction network for the drug-disease intersection targets was constructed by using Cytoscape 3.7.2 and the STRING database,and the core targets were screened.GO and KEGG pathway enrichment analyses of the core target genes were performed using the DAVID database and the bioinformatics platform.By establishing an Hp-positive perioral acne rat model through gastric gavage with Hp bacterial liquid and intradermal injection of Propionibacterium acnes suspension,the intervention effect of Dayuan Decoction was observed and the core targets and related pathways were preliminary validated.Morphology in the perioral skin and gastric tissue were observed through HE staining.The expressions of TLR4,MyD88,NF-κB in perioral skin tissue,the contents of TNF-α and IL-6 in serum were measured using ELISA.Results A total of 145 active components were screened from Dayuan Decoction,corresponding to 253 targets,with 57 targets overlapping between drugs and diseases.The core targets for the treatment of Hp-positive perioral acne with Dayuan Decoction were identified as IL6,TNF,IL1B,AKT1,TP53,etc.KEGG pathway enrichment analysis revealed 153 signaling pathways,primarily involved in pathways in cancer,lipids and atherosclerosis,AGE-RAGE signaling pathway in diabetic complications,TLR signaling pathway and TNF signaling pathway.Dayuan Decoction attenuated the expression of TLR4,MyD88 and NF-κB in the perioral skin of the Hp-positive perioral acne rat model,as well as reduced the contents of TNF-α and IL-6 in serum(P<0.01).Conclusion Dayuan Decoction may exert therapeutic effects on Hp-positive perioral acne by modulating the TLR4/NF-κB signaling pathway,reducing the levels of inflammatory factors TNF-α and IL-6,and alleviating the inflammatory damage in the perioral skin.