OBJECTIVES: To investigate the therapeutic mechanism of Thesium chinense Turcz. (TCT) for antibiotic-associated diarrhea (AAD). METHODS: Network pharmacology, KEGG pathway enrichment analysis and molecular docking were used to identify the shared targets and genes of TCT and AAD, the key signaling pathways and the binding between the active components in TCT and the core protein targets. In a Kunming mouse model of AAD established by intragastric administration of lincomycin hydrochloride, the effects of daily gavage of 1% carboxymethyl cellulose sodium or TCT gel solutions at 1.5 g/kg and 3 g/kg (n=10) on body weight and diarrhea were observed. HE staining, ELISA, 16S rRNA sequencing, and Western blotting were used to examine pathologies, expression levels of IL-6 and TNF-α, changes in gut microbiota, and protein expressions of EGFR, p-EGFR, PI3K, p-PI3K, Akt, and p-Akt in the colon tissues of the mice. RESULTS: We identified a total of 66 active components of TCT and 68 core targets including EGFR, STAT3 and PIK3CA. KEGG pathway enrichment analysis suggested that the therapeutic effects of TCT was mediated primarily through the PI3K/Akt signaling pathway. Molecular docking showed that EGFR had the highest binding affinity with coniferin, and the EGFR-coniferin complex maintained a stable conformation at 10 ns, whose stability was also confirmed by Gibbs free energy analysis. In the mouse models of AAD, treatment with TCT significantly improved colonic tissue morphology, decreased colonic levels of TNF-α and IL-6, increased gut microbiota diversity, and modulated the relative abundances of the key genera including Lactobacillus and Bacteroides. TCT treatment also markedly reduced protein expressions of p-EGFR, p-PI3K and p-Akt in the colon tissues of the mice. CONCLUSIONS TCT can alleviate AAD in mice by modulating gut microbiota composition, regulating the EGFR/PI3K/Akt signaling pathway, and reducing TNF‑α and IL-6 expressions.
Animals ; Gastrointestinal Microbiome/drug effects* ; Signal Transduction/drug effects* ; Mice ; ErbB Receptors/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Diarrhea/drug therapy* ; Phosphatidylinositol 3-Kinases/metabolism* ; Anti-Bacterial Agents/adverse effects* ; Drugs, Chinese Herbal/therapeutic use* ; Molecular Docking Simulation

OBJECTIVE: To analyze the clinical characteristics and genotypic changes of six children with RARS2 gene variants. METHODS: The clinical data of 6 children with RARS2 gene variants diagnosed at the Third Affiliated Hospital of Zhengzhou University from January 2017 to August 2024 were collected. Genetic variants were detected using trio-whole exome sequencing. Genomic DNA was extracted from samples and subjected to high-throughput sequencing. Variants were detected and analyzed using relevant databases and software. Pathogenic variants were validated by Sanger sequencing. The protein structure encoded by a previously unreported variant was predicted using a SWISS-MODEL online server. This study was approved by the Medical Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (Ethics No.: 2024-373-01). RESULTS: Among the six children, four were males and two were females, with the most recent follow-up age ranging from 1-year-and-1-month to 7 years old. The age of onset was under 1 year in all cases. All six children exhibited seizures, including infantile spasms in three, spasms and tonic spasms in one, and focal seizures in two. One child became seizure-free for 4 ~ 5 years following Valproic acid combined with topiramate and adrenocorticotropic hormone (ACTH) pulse therapy, but subsequently experienced a relapse. Another child has remained seizure-free for nearly one year with oral sodium valproate, levetiracetam, and a "cocktail" therapy. Seizures were not controlled in the remaining four children. Pontocerebellar hypoplasia was observed on neuroimaging in two children. All six patients exhibited severe psychomotor retardation. A total of 10 RARS2 gene variants were identified, three of which were previously unreported. CONCLUSION The predominant clinical features of Pontocerebellar hypoplasia associated with RARS2 gene variants include infantile onset, severe psychomotor retardation or regression, drug-resistant epilepsy, and feeding difficulties. The characteristic neuroimaging finding is pontocerebellar hypoplasia. However, its appearance may vary widely with time. The majority of affected children have a poor prognosis.
Humans ; Male ; Female ; Child, Preschool ; Infant ; Child ; Olivopontocerebellar Atrophies/genetics* ; Arginine-tRNA Ligase/genetics* ; Mutation ; Cerebellar Diseases

Objective:To analyze the clinical characteristics and genotypic changes of six children with RARS2 gene variants. Methods:The clinical data of 6 children with RARS2 gene variants diagnosed at the Third Affiliated Hospital of Zhengzhou University from January 2017 to August 2024 were collected. Genetic variants were detected using trio-whole exome sequencing. Genomic DNA was extracted from samples and subjected to high-throughput sequencing. Variants were detected and analyzed using relevant databases and software. Pathogenic variants were validated by Sanger sequencing. The protein structure encoded by a previously unreported variant was predicted using a SWISS-MODEL online server. This study was approved by the Medical Ethics Committee of the Third Affiliated Hospital of Zhengzhou University (Ethics No.: 2024-373-01). Results:Among the six children, four were males and two were females, with the most recent follow-up age ranging from 1-year-and-1-month to 7 years old. The age of onset was under 1 year in all cases. All six children exhibited seizures, including infantile spasms in three, spasms and tonic spasms in one, and focal seizures in two. One child became seizure-free for 4 ~ 5 years following Valproic acid combined with topiramate and adrenocorticotropic hormone (ACTH) pulse therapy, but subsequently experienced a relapse. Another child has remained seizure-free for nearly one year with oral sodium valproate, levetiracetam, and a " cocktail" therapy. Seizures were not controlled in the remaining four children. Pontocerebellar hypoplasia was observed on neuroimaging in two children. All six patients exhibited severe psychomotor retardation. A total of 10 RARS2 gene variants were identified, three of which were previously unreported. Conclusion:The predominant clinical features of Pontocerebellar hypoplasia associated with RARS2 gene variants include infantile onset, severe psychomotor retardation or regression, drug-resistant epilepsy, and feeding difficulties. The characteristic neuroimaging finding is pontocerebellar hypoplasia. However, its appearance may vary widely with time. The majority of affected children have a poor prognosis.

Objective This study sought to establish a non-alcoholic fatty liver disease(NAFLD)model in Wistar-SD hypercholesterolemia(WSHc)rats induced by a high-fat diet and to reveal the pathogenesis of NAFLD in these rats through the Srebp-1 gene.Methods After 2 weeks of dietary treatment,thirty 6-week-old WSHc rats were divided into High-fat control group,HFD+AAV no load group,and HFD+AAV group,with 10 rats in each group.The HFD+AAV no load group and HFD+AAV group were intravenously injected with a vector virus and an shRNA-containing virus,respectively.WSHc rats were fed with a normal fat diet as a normal control group.Serum levels of ALT,AST,TBIL,ALP,TBA,GLU,CHOL,and TG were measured every 2 weeks.After a further 8 weeks of feeding,the rats were euthanized and livers were excised for HE staining,Oil Red O staining,Masson staining,and Sirius red staining to observe the morphology,lipid deposition,and fibrosis of the liver tissues.RT-qPCR was performed to detect the expression of lipid metabolism-related genes namely Srebp-1,Aacs,FASN and LDLR in the livers.Furthermore,hepatocytes were isolated,cultured,and divided into a normal control group and a high-fat control group.Next,expression of the Srebp-1 gene was detected by RT-qPCR.Srebp-1 knockout(KO)hepatocytes were constructed,then TG content was detected and the lipid accumulation was observed by Oil Red O staining.Results After 10 weeks of high-fat diet treatment,serum ALT(P＜0.001),ALP(P＜0.001),TBA(P＜0.05),GLU(P＜0.001),and CHOL(P＜0.001)significantly increased in WSHc rats.Abnormal lipid deposition with formation of large vacuolar lipid droplets and fibrotic lesions in livers were observed.The mRNA expression of Srebp-1 noticeably increased in WSHc rats(P＜0.001).Moreover,compared with the high-fat control group,the ALT(P＜0.05)and GLU(P＜0.01)in the HFD+AAV group decreased,and liver lipid deposition and the formation of large vacuolar lipid droplets were alleviated.Expressions of genes such as FASN(P＜0.05)and LDLR(P＜0.01)were significantly upregulated.Additionally,there was a significant increase in the expression of Srebp-1 in hepatocytes of the high-fat control group(P＜0.001),while after Srebp-1 gene knockout,cellular TG levels decreased and the degree of lipid droplet aggregation was reduced.Conclusions The Srebp-1 gene plays a regulatory role in hepatic lipid metabolism and deposition,modulating the expression of lipid metabolism-related genes in WSHc rats with NAFLD.In vitro experiments demonstrated that downregulation of Srebp-1 alleviates lipotoxic injury in hepatocytes,suggesting that the development of NAFLD in WSHc rats is closely associated with abnormally high expressions of the Srebp-1 gene.

Objective To investigate changes in the pro-inflammatory mediator S100A8/9 and NLRP3/Caspase-1/IL-1β pathway in a rat kidney-aging model induced by D-galactose.Methods Twelve SD rats were divided into control and D-galactose groups,and injected subcutaneously in the back of the neck with D-galactose(150 mg/kg)to establish a rat model of kidney aging.Kidney samples were collected under anesthesia after 8 weeks.Kidneys were stained for senescence-associated beta-galactosidase(SA-β-Gal),mRNA expression levels of the aging-related genes p21,p16,and p53 were detected by quantitative reverse transcription-polymerase chain reaction(qRT-PCR),and histopathological changes were observed by hematoxylin-eosin(HE)and Masson staining.Serum urea nitrogen and creatinine,and catalase(CAT),glutathione peroxidase(GSH-PX),superoxide dismutase(SOD),and malondialdehyde(MDA)levels in the kidney tissues were detected.Reactive oxygen species(ROS)were detected by dihydroethdium staining and protein expression levels of collagen Ⅲ,α-smooth muscle actin(α-SMA),Protein expression of S100A8/9 was detected by immunofluorescence,and transforming growth factor(TGF)-β1 levels in kidney tissues and key factors in the NLRP3/Caspase-1/IL-1β inflammatory pathway were detected by Western Blot.A renal senescence model using HK-2 cells was constructed using H2O2 in vitro,and expression levels of the senescence proteins p21 and p16 and mRNA expression levels of the inflammatory factors IL-18 and tumor necrosis factor-α(TNF-α)were detected.Cell senescence was observed by SA-β-Gal staining.The effects of the S100A8/9 inhibitor paquinimod on expression levels of S100A8/9 and NLRP3/Caspase-1/IL-1β pathway-related proteins in the aging model were also detected.Results mRNA levels of the aging genes p21,p16,and p53 in kidney tissues were significantly increased in rats in the D-galactose group compared with the control group(P＜0.01),and SA-β-Gal staining showed a significant increase in senescent cells(P＜0.01).Serum blood urea nitrogen and creatinine levels increased(P＜0.05),CAT,GSH-PX,and SOD activities decreased(P＜0.01),while MDA activity increased in the D-galactose group(P＜0.01).Collagen Ⅲ,α-SMA,and TGFβ1 expression and the ROS content in tissues increased(P＜0.05).Glomeruli were atrophied or absent in the D-galactose group,the lumens of the renal sacs and renal tubules were enlarged,the nuclei were deeply stained and constricted,and numerous collagen fibers were deposited.Levels of S100A8 and S100A9 protein(P＜0.01),as well as NLRP3,Caspase-1,and IL-1β increased(P＜0.05).Paquinimod alleviated HK-2 cell senescence and decreased expression levels of the senescence proteins p21 and p16,and mRNA levels of the inflammatory factors IL-18 and TNF-α(P＜0.05,P＜0.01).The number of senile cells was also decreased,shown by SA-β-Gal staining(P＜0.01).Paquinimod also inhibited the protein expression of S100A8 and S100A9(P＜0.01)and NLRP3,Caspase-1,and IL-1β(P＜0.05 or P＜0.01).Conclusions S100A8/9 participates in the chronic inflammatory response by activating the NLRP3/Caspase-1/IL-1β pathway,thereby promoting D-galactose-induced renal aging.

Objective To investigate the therapeutic mechanism of Thesium chinense Turcz.(TCT)for antibiotic-associated diarrhea(AAD).Methods Network pharmacology,KEGG pathway enrichment analysis and molecular docking were used to identify the shared targets and genes of TCT and AAD,the key signaling pathways and the binding between the active components in TCT and the core protein targets.In a Kunming mouse model of AAD established by intragastric administration of lincomycin hydrochloride,the effects of daily gavage of 1%carboxymethyl cellulose sodium or TCT gel solutions at 1.5 g/kg and 3 g/kg(n=10)on body weight and diarrhea were observed.HE staining,ELISA,16S rRNA sequencing,and Western blotting were used to examine pathologies,expression levels of IL-6 and TNF-α,changes in gut microbiota,and protein expressions of EGFR,p-EGFR,PI3K,p-PI3K,Akt,and p-Akt in the colon tissues of the mice.Results We identified a total of 66 active components of TCT and 68 core targets including EGFR,STAT3 and PIK3CA.KEGG pathway enrichment analysis suggested that the therapeutic effects of TCT was mediated primarily through the PI3K/Akt signaling pathway.Molecular docking showed that EGFR had the highest binding affinity with coniferin,and the EGFR-coniferin complex maintained a stable conformation at 10 ns,whose stability was also confirmed by Gibbs free energy analysis.In the mouse models of AAD,treatment with TCT significantly improved colonic tissue morphology,decreased colonic levels of TNF-α and IL-6,increased gut microbiota diversity,and modulated the relative abundances of the key genera including Lactobacillus and Bacteroides.TCT treatment also markedly reduced protein expressions of p-EGFR,p-PI3K and p-Akt in the colon tissues of the mice.Conclusion TCT can alleviate AAD in mice by modulating gut microbiota composition,regulating the EGFR/PI3K/Akt signaling pathway,and reducing TNF-α and IL-6 expressions.

Objective:To explore the differences in sociodemographic and clinical characteristics between pa-tients with unipolar depression bipolar depression and to establish a nomogram for identifying bipolar depression.Methods:Using data from the China Mental Disorders Cohort Study,the sociodemographic and clinical characteristics of 2 643 patients with unipolar depression and 250 patients with bipolar depression diagnosed accord-ing to the criteria of the Diagnostic and Statistical Manual of Mental Disorders,Fifth Edition(DSM-5)were includ-ed to compare their sociodemographic and clinical characteristics.These characteristics included general demograph-ic information,disease-related information,clinical examination results,and the severity of the disease assessed with the Global Assessment of Functioning(GAF)and Hamilton Depression Rating Scale.Logistic regression analysis was employed to identify factors influencing bipolar depression,and a nomogram was constructed for its identifica-tion.Results:The risk factors for bipolar depression included being male(OR=1.48),being employed(OR=1.38),having non-melancholic features during episodes(OR=2.33),a Body Mass Index ranging from normal to obese(OR=2.48,2.49,4.65),psychotic features(OR=2.14),mixed episode(OR=9.36),comorbid physical diseases(OR=2.47),four or more depressive episodes(OR=1.67),earlier age of onset(OR=0.95),longer ill-ness duration(OR=1.03),and higher GAF scores(OR=1.02).The nomogram model achieved an AUC of 0.81(95％CI:0.78-0.84).The Hosmer-Lemeshow test result was x2=6.96(P＞0.05),indicating good model fit.The calibration curve showed good performance.The decision curve analysis revealed that the nomogram pro-vides significant clinical benefit when the risk of bipolar depression was within the range of 0 to 0.9.Conclusion:The nomogram established based on the identified sociodemographic and clinical factors can accurately assess the risk of bipolar depression,providing a useful tool for early identification and intervention.

Objective:To describe the time trend of suicide rate in China from 2016 to 2021,and to identify key populations and regions for targeted prevention.Methods:The crude suicide rates were obtained from China Health Statistical Yearbook,and the standardized suicide rate was calculated based on Population Census of China as the standard population.Poisson regression model was used to test the time trend of the average annual rate of sui-cide rate.The average suicide rates(average annual percentage change,AAPC)were calculated to describe suicide rates and trend in China.Results:From 2016 to 2021,the crude suicide rate(AAPC=-1.4％,95％CI:-3.6％-0.8％)and the standardized suicide rate(AAPC=-0.7％,95％CI:-2.5％-1.2％)showed a decreasing trend,but no statistically significance was found.Crude suicide rate in urban residents was lower than that in rural resi-dents.Crude suicide rate of the male was higher than that of the female in both urban and rural regions.The suicide rate of adolescents(AAPC=7.7％-25.2％),urban men aged 35-39 years(AAPC=3.4％)and men aged55-59 years(AAPC=2.1％-2.2％)in urban and rural regions had significantly increasing trends.The average annu-al suicide rate from 2016 to 2021 was 6.03/100 000,with a trend of increasing with age.Rural residents and men had higher average annual suicide rate.Conclusion:While overall suicide rates showed declining trends from 2016 to 2021,rising rates among adolescents and middle-aged men warrant targeted intervention.Continued efforts are needed to prevent suicide in rural areas and in old population.

Objective:To develop a Problematic Short Video Use Scale(PSVUS)and evaluate its validity and reliability.Methods:Through literature review and interviews,and with reference to the Diagnostic and Statisti-cal Manual of Mental Disorders,Fifth Edition(DSM-5)diagnostic guidelines for internet gaming disorder,the items were compiled.Initially,979 individuals(aged 14-74 years)participated in the survey,and 362 of them were re-tested after two weeks.The Perceived Stress Scale(PSS-4),UCLA Loneliness Scale(ULS-8),Brief Self-Control Scale(BSCS),and Hospital Anxiety and Depression Scale(HADS)were used to test the criterion-related validi-ty.Multiple classification methods were used to determine the cut-off value.Additionally,a 9-item brief version of the scale(PSVUS-9)was developed.Results:The PSVUS consists of 27 items organized into 9 dimensions(preoc-cupation,with drawal,tolerance,unsuccessful attempts,escape,continuation despite harm,deception,loss of inter-est,and functional impairment),explaining 78.71％of the total variance,with item factor loadings ranging from 0.62 to 0.95.The nine-factor structure exhibited good fit(x2/df=2.00,CFI=0.97,RMSEA=0.05).The scores of the dimensions were correlated with the scores of the PSS-4,ULS-8,BSCS and HADS(ICC=|0.21|-|0.68|,Ps＜0.05).The Cronbach's α coefficient of the PSVUS was 0.95,and the test-retest reliability was 0.83.The cut-off score for problematic use was determineded to be 70 or above.The PSVUS-9 score was correlated with the scores of the criterion measures(ICC=|0.39|-|0.68|,Ps＜0.01).The Cronbach's α coefficient of PSVUS-9 was 0.86 and the test-retest reliability was 0.80.A score of 24 or higher on the PSVUS-9 was indicative of problematic use.Conclusion:The Problematic Short Video Use Scale(PSVUS)and its brief version demonstrate good validity and reliability,and could be used to measure and screen for problematic short video use.

Objective:To develop and validate a combined model integrating radiomics, dosiomics, and clinical parameters based on CT simulation and dosimetric images in order to predict the occurrence of radiation pneumonitis (RP) in patients with non-small cell lung cancer (NSCLC).Methods:A retrospective study was conducted on the clinic data of 143 NSCLC patients who received radiotherapy at the Affiliated Hospital of Jining Medical University from January 2016 to December 2022. Patients were randomly stratified into a training group ( n = 100) and an internal validation group ( n = 43) at a 7∶3 ratio. Moreover, clinic data were collected from 34 NSCLC patients who received radiotherapy at the Jining Cancer Hospital between January 2019 and December 2022 as an external validation group. All three groups (the training group, internal validation, and external validation groups) were further categorized into two groups based on the RP severity (i.e., RP ≥ grade 2 and RP < grade 2). Their radiotherapy dose, CT simulation, and 3D dose distribution images were collected. Then, the total lung minus planning target volume (TL-PTV) was defined as the region of interest (ROI) for radiomics and dosiomic feature extraction, followed by feature dimensionality reduction. Consequently, key features associated with RP were determined. Four predictive models were developed using machine learning approaches (especially multilayer perceptron, MLP): a clinical model (CM), a radiomics model (RM), a dosiomics model (DM), and a radiomics and dosiomics nomogram (RDN), with a nomogram subsequently constructed. Ultimately, the performance and clinical feasibility of these models were assessed using receiver operating characteristic (ROC), area under the curve (AUC), and decision curve analysis (DCA). Results:A total of 1 834 radiomic features and 1 834 dosiomic features were extracted. Using the occurrence of RP ≥ grade 2 as the marker variable, 14 radiomic features, 15 dosiomic features, and three clinical features were selected from the training group to construct the prediction models (CM, RM, DM, and RDN). The performance and generalizability of these models were subsequently validated in both the internal validation and external validation groups. Specifically, the RDN exhibited AUCs of 0.915 (95% CI: 0.852-0.978), 0.879 (95% CI: 0.777-0.982), and 0.838 (95% CI: 0.701-0.975) in the three groups, respectively. A nomogram was established for RDN by integrating the radiomics score (R-score), dosiomics score (D-score), mean lung dose (MLD), V20, and V30. This nomogram allowed for individualized risk estimation of RP and facilitated personalized radiotherapy planning. Conclusions:The RDN model that is developed based on CT simulation and 3D dose distribution images and integrates radiomics, dosiomics, and clinical features can effectively predict the RP risk of NSCLC patients. The integration of multidimensional data contributes to the formation of the optimal predictive model, offering guidance for clinicians.