Objective This study aimed to explore tRNA-derived small RNAs(tsRNAs)associated with cardiac fibrosis and to investigate their regulatory mechanisms and clinical relevance. Methods We conducted a screening to identify tsRNAs relevant to cardiac fibrosis via PANDORA-seq and identified a significant increase in the expression of 5′tiRNA-His-GTG.We used qRT-PCR,Western blotting,CCK-8,and EdU to assess the effects of 5′tiRNA-His-GTG on the progression of cardiac fibrosis.Additionally,we utilized RNA pulldown and RIP-qPCR to explore the regulatory mechanism of 5′tiRNA-His-GTG in the TGF-β1/Smad3 pathway.We measured the expression level of 5′tiRNA-His-GTG in plasma samples from clinical patients and applied multivariate logistic regression analysis to determine whether 5′tiRNA-His-GTG is an independent risk factor for coronary atherosclerotic heart disease(CAD). Results Overexpression of 5′tiRNA-His-GTG promoted the fibrotic response in cardiac fibroblasts,increased the phosphorylation of Smad3 in the TGF-β1/Smad3 pathway,and activated the transcription and translation of downstream fibrosis-related genes,including COL1A1 and ACTA2.Mechanistically,5′tiRNA-His-GTG interacted with Smad3.The plasma levels of 5′tiRNA-His-GTG in the CAD group were approximately four fold higher than those in the control group,and 5′tiRNA-His-GTG was independently correlated with CAD onset. Conclusion Our results demonstrate that 5′tiRNA-His-GTG promotes Smad3 phosphorylation by interacting with Smad3,thus modulating the progression of cardiac fibrosis.

Objective To evaluate the clinical efficacy,safety,and mechanism of action of Bushen Ziyin Guchong decoction combined with moxibustion and auricular point pressing in the treatment of menorrhagia of kidney yin deficiency type. Methods A randomized controlled trial was conducted in which 138 menorrhagia patients of kidney yin deficiency type were enrolled.The participants were divided into an experimental group(69 cases)and a control group(69 cases),with a 3-menstrual cycle intervention.The experimental group received oral Bushen Ziyin Guchong decoction plus moxibustion at Yinbai point(SP1)and auricular point pressing.The control group was orally administered drospirenone or ethinylestradiol Table Ⅱ.Clinical efficacy,sex hormones(FSH,LH,E2,PRL,P,T),angiogenic factors(VEGF,ANG-1),endometrial thickness,and safety indicators were compared between the two groups. Results The total effective rate of the experimental group was significantly higher than that of the control group(95.70% vs.84.10%,P<0.05).The experimental group outperformed the control group in reducing the levels of FSH,LH,E2,P,VEGF,and ANG-1;improving PBAC scores(99.44±47.20 vs.116.55±49.98);decreasing endometrial thickness(0.51±0.24 vs.0.64±0.26 cm);and lowering TCM syndrome scores(10.36±3.38 vs.12.42±1.37 points)(all P<0.05).The incidence of adverse reactions was lower in the experimental group(5.8% vs.17.4%,P=0.033). Conclusion Bushen Ziyin Guchong decoction combined with moxibustion and auricular point pressing significantly improves menorrhagia of kidney-yin deficiency type by regulating hypothalamic-pituitary-ovarian(HPO)axis function,inhibiting pathological angiogenesis,and repairing the endometrial structure.Compared with single Western medicine,it has superior efficacy and better safety.

Objective To investigate the effects of remimazolam on airway inflammation and oxidative stress in a mouse model of ovalbumin(OVA)-induced bronchial asthma and explore the potential underlying mechanism of this effect. Methods Forty specific pathogen-free(SPF)male BALB/c mice(6-8 weeks old,18-20 g)were randomly divided into 4 groups(n=10 per group):control group,OVA-induced asthma group(OVA group),remimazolam intervention group(OVA+RMZL group),and dexamethasone(DEX)positive control group(OVA+DEX group).The bronchial asthma model was established via OVA sensitization followed by OVA challenge.One hour prior to each challenge,the mice in the OVA+RMZL group received an intraperitoneal injection of remimazolam(10 mg/kg),whereas the mice in the control group were administered phosphate-buffered saline(PBS,pH 7.4)instead.Airway pressure was measured via a lung function analyzer.Hematoxylin-eosin(HE)staining and periodic acid-Schiff(PAS)staining were performed to assess pathological changes and mucus secretion in lung tissues,respectively.An enzyme-linked immunosorbent assay(ELISA)was used to determine the concentrations of interleukin(IL)-4,IL-5,IL-13,and interferon-γ(IFN-γ)in bronchoalveolar lavage fluid(BALF),and OVA-specific immunoglobulin E(IgE)in serum.The levels of malondialdehyde(MDA),superoxide dismutase(SOD),and reduced glutathione(GSH)in lung tissues were detected via commercial assay kits.Reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting were employed to measure the mRNA and protein expression levels of nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase-1(HO-1),respectively,in lung tissues.Additionally,immunohistochemistry was used to evaluate the nuclear translocation of Nrf2 in lung tissues. Results Compared with the control group,the OVA group presented a significant increase in airway pressure,scores of lung inflammation and mucus secretion,levels of T helper 2(Th2)cytokines(IL-4,IL-5,IL-13)in BALF,serum OVA-specific IgE concentration,contents of reactive oxygen species(ROS)and MDA,as well as the mRNA and protein expression levels of Nrf2,HO-1,and NAD(P)H:quinone oxidoreductase 1(NQO1)(all P<0.05).In contrast,the OVA group presented significant decreases in the BALF IFN-γ level,SOD activity,and GSH content(all P<0.05).Compared with the OVA group,both the OVA+RMZL and OVA+DEX groups presented significant reductions in airway pressure,scores of lung inflammation and mucus secretion,levels of Th2 cytokines(IL-4,IL-5,IL-13)in BALF,serum OVA-specific IgE concentration,and contents of ROS and MDA(all P<0.05).Conversely,these two intervention groups presented significant increases in the IFN-γ level,SOD activity,GSH content,and Nrf2,HO-1,and NQO1 mRNA and protein expression levels(all P<0.05). Conclusion Remimazolam can alleviate airway inflammation and ameliorate oxidative stress in the lung tissues of OVA-induced asthmatic mice.The protective effect of remimazolam may be attributed to its ability to activate the Nrf2/HO-1 signaling pathway.

Objective To explore the value of nuclear receptor-binding protein 1(NRBP1)in assessing the prognosis of non-small cell lung cancer(NSCLC)patients and analyze the related regulatory mechanisms through in vitro experiments. Methods Sixty-seven NSCLC patients who underwent surgical treatment at Wuhan Third Hospital from January 2015 to December 2017 were included.Immunohistochemical analysis was performed to assess NRBP1 expression in NSCLC tumor tissues and adjacent tissues.Its correlation with clinicopathological parameters was analyzed.Additionally,the Kaplan-Meier plotter database was used to evaluate the association between NRBP1 and the prognosis of NSCLC patients.SiRNA-mediated knockdown of NRBP1 expression in A549 and 95D cells was performed,and cell proliferation ability was detected via a CCK-8 assay.Cell colony formation experiments were performed to investigate the effects on colony formation.Flow cytometry was used to analyze changes in the cell cycle.qRT-PCR and Western blotting were employed to analyze NRBP1 expression.The expression of Ki-67,CDK4,cyclin D1,and P21,which are related to cell proliferation and the cell cycle in A549 cells,was detected in A549 cells via Western blotting. Results Compared with that in adjacent tissues,the positive expression rate of NRBP1 in NSCLC lung cancer tissue was greater(P<0.01),which was correlated with TNM stage(P<0.05).Compared with patients with high NRBP1 expression,patients with low NRBP1 expression had longer overall survival[HR=1.28(1.13-1.44),log-rank P<0.01].Downregulation of NRBP1 expression in A549 cells and 95D cells inhibited cell proliferation and colony formation,increasing the number of cells in the G₁ phase and decreasing the number of cells in the S phase.Downregulation of NRBP1 expression in A549 cells led to decreases in Ki-67,CDK4,and cyclin D1 expression and an increase in P21 expression. Conclusion Increased expression of NRBP1 in NSCLC is associated with poor prognosis,and NRBP1 can participate in the regulation of NSCLC cell proliferation by altering the G₁/S phase of the cell cycle.

Objective By integrating multi-omics and clinical information,we conducted a comprehensive analysis of specific alternative polyadenylation(APA)alterations and their prognostic significance in breast cancer(BRCA)and explored the key upstream and downstream regulatory factors involved. Methods Multiple omics datasets from the TCGA and GTEx cohorts were integrated to identify dysregulated APA events in BRCA.Clinical information and LASSO Cox regression analysis were used to identify APA events associated with prognosis.Potential upstream APA regulators and somatic mutations influencing prognostic APA events were identified through multivariate linear regression and LASSO regression analyses.Finally,changes in downstream miRNA binding sites associated with APA events were analyzed based on miRNA-3′untranslated regions(3′UTR) regulatory information. Results An in-depth and systematic analysis of APA profiles in BRCA revealed widespread APA dysregulation,with 77.37% of altered APA events involving transcript shortening,predominantly affecting cancer- and immune-related processes.A total of 112 prognosis-related APA events were identified,and an overall survival(OS)prediction signature was constructed on the basis of 10 core APA events.Further analysis pinpointed SNRNP70 and PABPN1 as crucial regulators of APA deregulation,with SNRNP70 downregulation potentially causing APA-mediated shortening of STARD10.Moreover,systematic analysis of APA-modulated somatic mutations revealed that they were associated with 82.44% of dysregulated APA events and may influence APA by altering gene function.The construction of an APA-miRNA network for BRCA further revealed that APA-oriented dysfunction in STARD10 arises from its evasion of miR-325-3p suppression. Conclusion This study identified a series of prognosis-related APA events and their critical upstream and downstream regulatory factors through an integrative multi-omics approach,providing insights into the regulatory mechanisms of APA in BRCA.

Objective This study aimed to investigate the therapeutic mechanism of Agrimonia pilosa in the treatment of bronchial asthma(BA) via network pharmacology and molecular docking techniques,followed by validation through animal experiments. Methods The key bioactive components and core therapeutic targets of Agrimonia pilosa for treating BA were identified from multiple databases,including TCMSP,OMIM,DisGenet,and GeneCards.The protein-protein interaction(PPI)network of these targets was subsequently constructed via STRING and Cytoscape,and the core targets were selected for Gene Ontology(GO)functional annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.A BA rat model was established by sensitizing the rats with 4% ovalbumin.The levels of IL-4,IL-5,IL-6,and IFN-γ in bronchoalveolar lavage fluid were determined via ELISA,while the protein expression levels of p38 MAPK,P-p38 MAPK,NF-κB p65,and NF-κB P-p65 in lung tissue were assessed via Western blotting to evaluate the effects of Agrimonia pilosa on BA rats and verify the associated signaling pathways. Results A total of five active components of Agrimonia pilosa were identified,along with 120 intersecting potential therapeutic targets for BA.Among these,five inflammation-related core targets,which were found to be involved in 159 signaling pathways,were highlighted.Molecular docking analysis revealed that quercetin displayed stable binding and high affinity with the identified inflammation-related core targets.The results of animal experiments using the BA model revealed that Agrimonia pilosa decreased the levels of the cytokines IL-4,IL-5,IL-6,and IL-13 while increasing the IFN-γ level.Moreover,Western blot analysis revealed that Agrimonia pilosa downregulated the protein expression of P-p38 MAPK and NF-κB P-p65. Conclusion Agrimonia pilosa may exert its effects by inhibiting the p38 MAPK/NF-κB signaling pathway,thereby reducing Th2 cytokine production and alleviating inflammatory responses,which may be a mechanism of Agrimonia pilosa in the treatment of BA.

Objective To evaluate the impact of the first-trimester systemic immune-inflammation index(SII)and systemic inflammation response index(SIRI)on gestational diabetes mellitus(GDM),subtypes of GDM,and blood glucose levels. Methods A retrospective cohort study was conducted,involving 34,171 pregnant women from the electronic medical record system of Guangdong Women and Children Hospital from 2015 to 2024.GDM patients and their subtypes were diagnosed via the 75 g oral glucose tolerance test.A modified Poisson regression model,multinomial Logistic regression model and linear regression model were used to explore the associations of the SII and SIRI with GDM,subtypes of GDM and blood glucose levels,respectively.A restricted cubic spline(RCS)model was applied to explore the possible nonlinear relationships between SII,SIRI and GDM as well as blood glucose levels. Results After adjusting for covariates,a modified Poisson regression model revealed that an elevated ln-transformed SII(RR=1.30,95%CI:1.23-1.37)and SIRI(RR=1.21,95%CI:1.16-1.27)were associated with an increased risk of developing GDM.The multinomial logistic regression model revealed that elevated ln-transformed SII and SIRI were associated with an increased risk of developing GDM-isolated post-load hyperglycemia(GDM-IPH)(SII:OR=1.36,95%CI:1.26-1.47;SIRI:OR=1.27,95%CI:1.19-1.36)and GDM-combined hyperglycemia(GDM-CH)(SII:OR = 1.82,95%CI:1.49-2.21;SIRI:OR = 1.47,95%CI:1.25-1.73),but elevated SII and SIRI did not have statistically significant effect on the risk of developing GDM-isolated fasting hyperglycemia(GDM-IFH)(SII:OR=1.22,95%CI:0.99-1.51;SIRI:OR=1.09,95%CI:0.92-1.30).The RCS results revealed that the ln-transformed SII exhibited a J-shaped dose-response relationship with fasting and 1-hour blood glucose levels and a linear dose-response relationship with 2-hour blood glucose levels,whereas the ln-transformed SIRI exhibited a linear dose-response relationship with 1- and 2-hour blood glucose levels. Conclusion Elevated SII and SIRI in the first trimester are significantly associated with an increased risk of GDM and its GDM-IPH and GDM-CH subtypes.The results provide new inflammatory markers for early risk identification and subtype-specific management of GDM.

Objective To investigate the possible mechanism by which G protein-coupled estrogen receptor(GPER)improves mitochondrial dysfunction in the hepatocytes of patients with sepsis. Methods An in vitro sepsis model was established by stimulating AML-12 hepatocytes with lipopolysaccharide(LPS).The cells were divided into four groups:control,LPS-treated,LPS + G1(a GPER agonist),and LPS + G15(a GPER antagonist)groups.Cell viability,apoptosis,the levels of inflammatory factors(TNF-α,IL-1β,and IL-6),and energy metabolism indicators(ATP/AMP)were assessed via techniques such as CCK-8,flow cytometry,and ELISA.The mRNA and protein expression levels of GPER were determined by RT-qPCR and Western blotting respectively.Western blot analysis was also employed to examine AMPK phosphorylation(p-AMPK)and the protein expression of pyruvate dehydrogenase E1 alpha subunit(PDHA1)and carnitine palmitoyltransferase 1B(CPT1B).Mitochondrial reactive oxygen species(mtROS)and mitochondrial membrane potential(ΔΨm)were measured via laser scanning confocal microscopy and flow cytometry,respectively.The ultrastructure of the mitochondria was observed via transmission electron microscopy. Results In the LPS-induced hepatocyte sepsis model,the GPER agonist G1 not only significantly improved cell viability,inhibited apoptosis,and reduced inflammatory responses but also effectively reversed energy metabolism disorders,decreased oxidative stress(all P<0.01),and repaired mitochondrial ultrastructural damage.In contrast,the GPER antagonist G15 not only completely abolished the protective effects of G1 but also further exacerbated these damage indicators(all P<0.01). Conclusion GPER protects against sepsis-induced hepatocyte injury via activating the AMPK-PDHA1/CPT1B axis,which enhances glucose and lipid metabolism,preserves mitochondrial energy homeostasis,and mitigates oxidative stress.These findings suggest that GPER is a potential novel target for sex-specific therapeutic strategies in sepsis.

Objective The effects of glycyrrhetinic acid on sorafenib-induced cardiomyocyte injury and apoptosis were investigated via the hypoxia-inducible factor-1α(HIF-1α)/adenovirus E1B 19 kDa-interacting protein 3(BNIP3)signaling pathway. Methods MTT was used to determine the effects of glycyrrhetinic acid on H9c2 cells in rat cardiomyocytes.The experiment was divided into control group,sorafenib group,sorafenib+glycyrrhetinic acid group,sorafenib+glycyrrhetinic acid+HIF-1α inhibitor YC-1 group,and sorafenib+glycyrrhetinic acid+YC-1+autophagy agonist rapamycin(RAPA)group.A kit was used to determine LDH release and MDA levels.The fluorescent probe method was used to determine the level of total ROS in the cells.Apoptosis was detected via flow cytometry.Immunofluorescence was used to detect the protein expression of LC3 and the colocalization of BNIP3 and LC3.The expression levels of the mitochondrial autophagy-related proteins LC3 II/I,p62 and Beclin-1 were detected via Western blotting.RT-qPCR and Western blotting were used to detect the mRNA and protein expression levels of HIF-1α and BNIP3,respectively. Results Compared with that in the sorafenib group,the viability of H9c2 cells in the 100 and 200 μmol/L glycyrrhetinic acid groups was significantly increased(P<0.05).Compared with those in the control group,LDH release and the MDA and ROS levels in H9c2 cells in the sorafenib group were significantly up-regulated(P<0.05);the apoptosis rate of cells was significantly increased(P<0.05);the expression levels of LC3 II/I and Beclin-1 protein were significantly down-regulated(P<0.05);and the levels of p62 were significantly up-regulated(P<0.05);the fluorescence intensity of LC3 in H9c2 cells was weakened,and the co-localization of LC3 and BNIP3 in H9c2 cells was reduced,the mRNA and protein expression levels of HIF-1α and BNIP3 were significantly down-regulated(P<0.05).Compared with those in the sorafenib group,LDH release and the MDA and ROS levels in H9c2 cells in the sorafenib+glycyrrhetinic acid group were significantly down-regulated(P<0.05);the apoptosis rate of H9c2 cells was significantly reduced(P<0.05);the expression levels of LC3 II/I and Beclin-1 proteins were significantly up-regulated(P<0.05);the p62 levels were significantly down-regulated(P<0.05);the fluorescence intensity of LC3 in H9c2 cells was enhanced,the colocalization of LC3 and BNIP3 in H9c2 cells was enhanced,the mRNA and protein expression levels of HIF-1α and BNIP3 were significantly up-regulated(P<0.05).YC-1 significantly reversed these effects(P<0.05),and RAPA significantly inhibited the effect of YC-1(P<0.05). Conclusion Glycyrrhetinic acid can promote mitophagy by activating the HIF-1α/BNIP3 signaling pathway and can inhibit sorafenib-induced cardiomyocyte injury and apoptosis.

Objective To investigate the effects of Decursin(DE)on the proliferation and migration of colorectal cancer cells and the underlying mechanism. Methods Third-generation SW480 and HCoEpiC cells were selected for the experiment,and the expression of miR-17-5p RNA in the two cell lines was detected via qRT-PCR.The proliferation of SW480 cells was detected via the CCK-8 method at different DE concentrations(0,10,30,60 and 100 μmol/L),and the appropriate concentration of DE was selected.qRT-PCR was used to detect the transfection efficiency of miR-17-5p in SW480 cells.The proliferation of the transfected cells was detected via CCK-8 method,and the number of migrating cells was detected via Transwell experiment.The expression of miR-17-5p was detected via qRT-PCR,the cell proliferation activity was detected via the CCK-8 method,the number of migrating cells was detected via Transwell experiment,and the levels of PIK3R1,p-PI3K,PI3K and p-Akt were detected via Western blotting.The dual-luciferase reporter assay was used to detect the targeting effect of miR-17-5p on PI3K. Results The expression level of miR-17-5p RNA in W480 cells was greater than that in HCoEpiC cells(P<0.05).The proliferation of SW480 cells decreased with increasing DE concentration in a concentration-dependent manner(P<0.05).For the subsequent experiments,60 μmol/L DE was selected.The miR-17-5p mimic/inhibitor significantly increased/decreased the expression of miR-17-5p,cell proliferation activity and the number of migrating cells(P<0.05).After treatment with DE,the expression of miR-17-5p,cell proliferation activity,cell migration,and p-PI3K and p-Akt proteins decreased(P<0.05),whereas the expression of the PIK3R1 protein increased(P<0.05);however,the expression of the PI3K and Akt proteins did not change significantly.When miR-17-5p and Wt-PIK3R1 were co-transfected,the relative luciferase activity decreased/increased with increasing/decreasing miR-17-5p expression(P<0.05).When miR-17-5p and Mut-Wt-PIK3R1 were co-transfected,the relative activity of luciferase did not change significantly(P> 0.05). Conclusion Decursin can inhibit the proliferation and migration of colorectal cancer cells,which may inhibit the PI3K/Akt pathway by downregulating miR-17-5p and promoting the expression of PIK3R1.

Objective To explore the regulatory role of mechanistic target of rapamycin complex 1(mTORC1)on the protein stability of triphenyl peptidase 1(TPP1)through β-TrCP. Methods Interactions between TPP1 and mTORC1 or β-TrCP were investigated via GST Pull-down,coimmunoprecipitation and Western blotting.The cells were cultured under various nutrient stress conditions or treated with rapamycin to inhibit mTORC1 activity.TPP1 protein expression was assessed via Western blotting,while TPP1 transcriptional levels were measured via qPCR.Proteasome or cullin neddylation inhibitors were used to determine whether TPP1 degradation is mediated by the proteasome and Cullin-dependent E3 ubiquitin ligases.Putative β-TrCP recognition phosphodegron motifs in TPP1 were mutated via site-directed mutagenesis,replacing serine(S)residues with alanine(A).Lentivirus-delivered shRNA was employed to silence β-TrCP or Cullin1 gene expression.TPP1 phosphorylation and ubiquitination levels were detected by Western blotting,and the essential role of two serine residues within the phosphodegron in its ubiquitination was functionally validated.In vitro TPP1 activity assays were performed to assess whether phosphodegron mutations affect TPP1 enzymatic activity. Results TPP1 interacted with both mTORC1 and β-TrCP.TPP1 protein levels were negatively correlated with mTORC1 activity,and this regulation was not due to transcriptional changes.Inhibition of the proteasome or knockdown of β-TrCP increased TPP1 protein levels.In vitro kinase and in vivo ubiquitination assays confirmed that TPP1 was phosphorylated by mTORC1 and ubiquitinated by β-TrCP.Mutations in the TPP1 phosphodegron did not affect its enzymatic activity. Conclusion mTORC1 phosphorylates TPP1,promoting its specific binding to β-TrCP,leading to TPP1 ubiquitination and subsequent proteasomal degradation.

Objective This study aimed to explore the protective effect of tocilizumab on acute myocardial injury induced by doxorubicin. Methods Human AC16 cardiomyocytes were treated with different concentrations of tocilizumab(1,3,and 5 μg/mL)before exposure to doxorubicin(1 μmol/L).Cell viability was assessed using the CCK-8 assay,while apoptosis was measured by flow cytometry.The ROS level was assessed with a DCFH2-DA probe.JC-1 staining was used to analyze the mitochondrial membrane potential.The expression levels of CXCL10,STING,and cGAS were detected through Western blotting and qRT-PCR. Results In the context of the cytotoxicity induced by doxorubicin,tocilizumab significantly increased AC16 cell viability(P<0.01)and reduced early and late apoptosis rates(P<0.01).In addition,tocilizumab reduced the ROS level(P<0.01)and downregulated the expression of cGAS,STING and CXCL10. Conclusion Tocilizumab has a cardioprotective effect on doxorubicin-induced myocardial injury by modulating the cGAS-STING pathway,highlighting its potential as a therapeutic agent to improve cardiac safety during chemotherapy.

Objective To investigate the expression level of LHPP in human breast cancer and its impact on proliferation and migration and to conduct a preliminary exploration of its immunoregulatory function. Methods The expression level of the LHPP protein was examined in breast cancer patients.To manipulate LHPP expression,breast cancer cells were transfected with overexpression plasmids (T47D) or specific shRNAs(MCF7).LHPP protein levels,along with cell proliferation,migration capacity,and Akt pathway activity,were quantified via Western blotting,CCK-8,Transwell,and related assays,respectively.A preliminary investigation was conducted to examine the role of LHPP in immune regulation. Results The expression was significantly lower in cancer tissues than in adjacent non-tumor tissues from breast cancer patients(P<0.05).LHPP expression varied across cancer cell lines,with T47D showing lower levels and MCF7 exhibiting relatively higher levels(all P<0.05).In T47D cells,LHPP overexpression successfully led to increased levels of LHPP.As a result,cell proliferation,migration,and the p-Akt/Akt ratio were significantly decreased(all P<0.05).In MCF7 cells,shLHPP transfection knocked down LHPP expression,leading to significant changes in the above-mentioned phenotypic and pathway markers(all P<0.05). Conclusion LHPP,which is expressed at low levels in breast cancer,functions as a tumor suppressor by inhibiting cancer cell proliferation and migration.This effect may be mediated through the Akt signaling pathway.

Objective To investigate the genetic etiology and analyze the pathogenicity of the detected mutations in three sporadic Marfan syndrome(MFS)patients without family history. Methods Three probands with typical MFS features but no family history who were admitted to Xijing Hospital between 2024 and 2025 were enrolled as subjects.A retrospective study was conducted to collect clinical data from these three families.Peripheral blood samples were collected from the probands and their available family members,and genomic DNA was extracted.Whole-exome sequencing(WES)was performed to screen for genetic variants.Sanger sequencing was used to validate the identified FBN1 gene variants in the probands and their family members.The pathogenicity of the detected variants was classified according to the Standards and Guidelines for the Interpretation of Sequence Variants established by the American College of Medical Genetics and Genomics(ACMG).The potential impact of the variants on protein structure was analyzed via the online tool Mutation Taster and Swiss-Pdb Viewer software. Results Clinical features are as follows:All three probands exhibited cardiovascular system abnormalities.Probands from families 1 and 2 presented ocular abnormalities,whereas all three probands presented skeletal system manifestations.Genetic testing results are as follows:Three different FBN1 gene variants involving three distinct exons were identified across the three MFS families,namely,c.3787T>G(p.Cys1263Gly),c.5965T>C(p.Cys1989Arg),and c.6610T>G(p.Cys2204Gly).Pathogenicity assessment results are as follows:According to the ACMG guidelines,all three variants were classified as“likely pathogenic”.Database and literature searches confirmed that these variants were unreported and thus novel.Parental screening verified that all three mutations were de novo.Bioinformatics analysis results are as follows:Predictions via SIFT and PolyPhen-2 software indicated that the c.3787T>G(p.Cys1263Gly),c.5965T>C(p.Cys1989Arg),and c.6610T>G(p.Cys2204Gly)variants were harmful.Amino acid sequence alignment revealed high conservation of the mutated residues across different species in the FBN1 protein.Comparative analysis of the three-dimensional protein structure revealed that the c.6610T>G(p.Cys2204Gly)variant altered hydrogen bonds in the vicinity of the amino acid,further supporting the association between FBN1 gene variation and MFS. Conclusion This study identified three novel FBN1 gene variants,expanding the mutational and phenotypic spectrum of MFS.Parental testing confirmed their de novo origin,revealing the etiology of sporadic MFS without family history.These findings provide a basis for genetic counseling,clinical diagnosis,and management.

Objective To explore the impact of creative game intervention on core symptoms and neuroimmune function in children with autism and to analyze the relationship between core symptoms and neuroimmune function. Methods A total of 100 children with autism who visited our hospital for treatment from February 2022 to February 2025 were enrolled and randomly divided into an observation group(receiving creative play intervention)and a control group(receiving conventional rehabilitation intervention),with 50 cases in each group.Core symptom indicators[Childhood Autism Rating Scale(CARS),Autism Treatment Evaluation Checklist(ATEC),Autism Behavior Checklist(ABC)]and neuroimmune indicators[interleukin-17A(IL-17A),anti-myelin basic protein antibody(anti-MBP),IL-6,and osteopontin(OPN)]were compared between the two groups before and after intervention.Pearson correlation analysis was used to examine the correlations between core symptom indicators and neuroimmune indicators in the observation group. Results Before intervention,there were no statistically significant differences in CARS scores,ATEC scores,or ABC scores between the two groups(all P>0.05).After the intervention,the CARS scores,ATEC scores,and ABC scores of both groups were significantly lower than those before the intervention,and the scores of the observation group were significantly lower than those of the control group(all P<0.05).Before intervention,there were no statistically significant differences in the levels of IL-17A,anti-MBP,IL-6,or OPN between the two groups(all P> 0.05).After intervention,the levels of IL-17A,anti-MBP,IL-6,and OPN in both groups were significantly lower than those before intervention,and the levels in the observation group were significantly lower than those in the control group(all P<0.05).In the observation group,IL-17A,anti-MBP,IL-6,and OPN were positively correlated with the CARS score(r=0.442, 0.479, 0.483, 0.343, respectively), ATEC score(r=0.547, 0.610, 0.585, 0.406, respectively), and ABC score(r=0.463, 0.496, 0.513, 0.417, respectively) (all P<0.05). Conclusion Creative game intervention is beneficial for improving core symptoms and neuroimmune dysfunction in children with autism,and it is more effective than conventional rehabilitation intervention.The core symptoms in children with autism are closely related to neuroimmune function.

Objective T o investigate the association between the triglyceride-glucose(TyG)index and mild cognitive impairment(MCI)in older adults. Methods Older adults aged ≥ 60 years who underwent health examinations in the Shuiguohu Community of Wuhan,Hubei Province,from September 2023 to September 2024 were enrolled.According to the inclusion and exclusion criteria,487 participants were ultimately included.General demographic characteristics and laboratory data were collected,and body mass index(BMI)and the TyG index were calculated.The participants were divided into an MCI group and a normal cognition group on the basis of the Montreal Cognitive Assessment(MoCA)score.Between-group comparisons were performed via independent-samples t tests,Wilcoxon rank-sum tests,and Fisher’s exact tests.After adjustment for confounders,including age,sex,and BMI,binary logistic regression was used to analyze the association between the TyG index and MCI,and subgroup analyses were conducted according to combined categories of age and BMI.Restricted cubic spline(RCS)regression was applied to evaluate the dose-response relationship between the TyG index and MCI. Results Among the 487 participants,203 were classified into the MCI group,and 284 into the normal group.Compared with that in the normal group,the TyG index was significantly increased in the MCI group(P<0.05).After adjustment for confounding factors,binary logistic regression revealed that the TyG index was positively associated with MCI(P= 0.036).RCS regression indicated a linear increasing trend in MCI risk with increasing TyG index values(P_overall= 0.038,P_non-linear> 0.05).Subgroup analysis on the basis of combined age and BMI data revealed that an elevated TyG index was significantly positively associated with MCI risk among individuals aged ≥70 years with a BMI ≥ 24 kg/m². Conclusion Elevated TyG index is associated with an increased risk of MCI,suggesting that the TyG index may serve as a potential indicator for the early identification of MCI.

Traditional animal models have inherent limitations in the genetic manipulation,developmental visualization,and high-throughput screening for intracranial aneurysms(IAs).The emergence of the zebrafish model has been expanding the experimental approaches and research platforms available in these fields.This review systematically analyzes the biological basis of using zebrafish to investigate human cerebrovascular diseases,summarizes several established zebrafish aneurysm models,and introduces their applications in studies of IA mechanisms.Unlike previous reviews that mainly compared the construction methods and strengths of different models in parallel,this paper also focuses on the applications of zebrafish aneurysm models in studying disease mechanisms,aiming to promote a broader application in IA pathogenesis research and drug screening.

Post-stroke cognitive impairment(PSCI)is a common sequela in stroke patients and significantly affects their quality of life and rehabilitation progress.As a green,economical,and safe therapeutic approach,acupuncture has been demonstrated to possess unique advantages in treating PSCI.Electroencephalography(EEG),a noninvasive neuroimaging technique,has become a focus of clinical research because of its ability to predict cognitive impairment.Studies investigating the mechanisms underlying the effects of acupuncture on PSCI via EEG technology have revealed that acupuncture can improve cognitivefunction by modulating brain electrical activity.The integration of acupuncture and EEG technology shows great potential and promising application prospects in the assessment and treatment of PSCI.

In recent years,the increasing prevalence and early onset of myopia have made its control a pressing public health priority.Orthokeratology(OK lenses)is regarded as one of the key effective strategies for myopia management,although its exact mechanisms of action remain incompletely understood.The predominant hypothesis is that orthokeratology modifies the relative optical power of the cornea,thereby inducing peripheral retinal defocus and associated choroidal changes,which together contribute to slowing axial elongation.Grounded in peripheral defocus theory,this article reviews alterations in retinal peripheral defocus and the choroidal response following orthokeratology,discusses methods for assessing retinal peripheral defocus,examines factors influencing the magnitude of defocus,and explores its correlation with myopia control outcomes.The objective of this study is to clarify the mechanism through which orthokeratology effectively mitigates myopia progression.

Peripheral neuropathies are a collection of disorders characterized by injury and generalized dysfunction of nerves.Pathogenic factors are complex,including trauma,hyperglycemia,side effects of chemotherapeutic drugs and inflammatory,immune-related,metabolic,endocrine-related,and neoplastic factors.In addition,it can cause great problems for patients.Oxidative stress,as one of the key causes of peripheral neuropathy,is characterized mainly by an imbalance between oxidative processes and antioxidant defenses in the body,which tends to oxidize,leading to inflammatory infiltration of neutrophils,increased protease secretion,and the production of many oxidative intermediates.At present,antioxidants are widely used in clinical practices to inhibit oxidative stress to improve peripheral neuropathy,but their efficacy is limited.Nuclear factor-erythroid 2-related factor 2(Nrf2),a key transcription factor of intracellular oxidative stress,mediates antioxidant effects through numerous signaling pathways,such as the Nrf2/HO-1,Nrf2/NQO1,PERK/Nrf2,Nrf2/ERK,and Nrf2/NF-κB pathways.In the present study,the oxidative stress pathways associated with the targeting of cellular Nrf2 factors were reviewed,hopefully providing evidence for peripheral neuropathy-related research.

Cardiac arrhythmia is one of the most common causes of death worldwide.Most antiarrhythmic drugs lack specificity and are associated with numerous adverse effects.More importantly,nearly all antiarrhythmic drugs possess potential arrhythmogenic properties,which may exacerbate existing arrhythmias or induce new ones.The glutamatergic and GABAergic systems,as novel endogenous regulatory mechanisms,confer distinct functional plasticity in cardiomyocytes.Elucidating the diverse biological roles of the glutamatergic and GABAergic systems in cardiac tissues,as well as identifying the conditions that lead to their dysregulation,is critical for the development of novel therapeutic strategies targeting this rhythm-regulating system in the heart.