A recently published study(Xin et al.,Prog Biochem Biophys,2026,53(2):431-441.DOI:10.3724/j.pibb.2025.0508)addresses the therapeutic challenges of pancreatic ductal adenocarcinoma(PDAC)by innovatively developing an or...A recently published study(Xin et al.,Prog Biochem Biophys,2026,53(2):431-441.DOI:10.3724/j.pibb.2025.0508)addresses the therapeutic challenges of pancreatic ductal adenocarcinoma(PDAC)by innovatively developing an orally administered nanogene delivery system.Designed to achieve in situ,efficient delivery of chimeric antigen receptor(CAR)genes to tumor sites,this approach offers a novel strategy for CAR-macrophage(CAR-M)based immunotherapy.Its key highlights are as follows.展开更多
Dianthus spiculifolius Schur,as an emerging ornamental plant,has extensive applications and economic values.In this study,the DsCBL4 gene was successfully cloned,and its tissue-specific expression,expression patterns ...Dianthus spiculifolius Schur,as an emerging ornamental plant,has extensive applications and economic values.In this study,the DsCBL4 gene was successfully cloned,and its tissue-specific expression,expression patterns under various abiotic stresses,subcellular localization,and bioinformatics analysis of the encoded amino acid sequence were conducted.The results showed that the coding region of the DsCBL4 gene was 675 bp long,encoding 224 amino acids.It had high homology with the amino acids encoded by Amaranthus tricolor,Chenopodium quinoa and Spinacia oleracea.The predicted relative molecular mass of DsCBL4 was 25.61 ku,with an isoelectric point of 4.58,and it had phosphorylation sites,belonging to an unstable hydrophilic protein.Its secondary structure includedα-helices,irregular coils and extended chains.The tertiary structure prediction revealed that DsCBL4 had four EFhand calcium-binding domains necessary for Ca2+binding in plant calmodulin-like proteins and the FPSF motif for calcineurin B-like protein(CBL)-interacting protein kinase(CIPK)activation.The expression level of the DsCBL4 gene showed tissue specificity,with the highest expression in roots.It was induced by drought,low temperature,combined drought and low temperature,salt stress,nitrogen stress,phosphorus stress,calcium ion stress,high temperature stress,and abscisic acid(ABA)stress.Both transient infection in Nicotiana tabacum L.and stable expression in transgenic Arabidopsis thaliana showed that the DsCBL4 protein was localized to the cell membrane.These results suggested that DsCBL4 might be involved in the abiotic stress response of Dianthus spiculifolius through the calcium signaling pathway,providing a theoretical basis for understanding its molecular mechanism.This study provided an important reference for further exploring the role of the DsCBLs gene family in plant stress resistance.展开更多
Objective:Osteoarthritis(OA)and sarcopenia are significant health concerns in the elderly,substantially impacting their daily activities and quality of life.However,the relationship between them remains poorly underst...Objective:Osteoarthritis(OA)and sarcopenia are significant health concerns in the elderly,substantially impacting their daily activities and quality of life.However,the relationship between them remains poorly understood.This study aims to uncover common biomarkers and pathways associated with both OA and sarcopenia.Methods:Gene expression profiles related to OA and sarcopenia were retrieved from the Gene Expression Omnibus(GEO)database.Differentially expressed genes(DEGs)between disease and control groups were identified using R software.Common DEGs were extracted via Venn diagram analysis.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were conducted to identify biological processes and pathways associated with shared DEGs.Protein-protein interaction(PPI)networks were constructed,and candidate hub genes were ranked using the maximal clique centrality(MCC)algorithm.Further validation of hub gene expression was performed using 2 independent datasets.Receiver operating characteristic(ROC)curve analysis was used to evaluate the predictive value of key genes for OA and sarcopenia.Mouse models of OA and sarcopenia were established.Hematoxylin-eosin and Safranin O/Fast Green staining were used to validate the OA model.The sarcopenia model was validated via rotarod testing and quadriceps muscle mass measurement.Real-time reverse transcription PCR(real-time RT-PCR)was employed to assess the mRNA expression levels of candidate key genes in both models.Gene set enrichment analysis(GSEA)was conducted to identify pathways associated with the selected shared key genes in both diseases.Results:A total of 89 common DEGs were identified in the gene expression profiles of OA and sarcopenia,including 76 upregulated and 13 downregulated genes.These 89 DEGs were significantly enriched in protein digestion and absorption,the PI3K-Akt signaling pathway,and extracellular matrix-receptor interaction.PPI network analysis and MCC algorithm analysis of the 89 common DEGs identified the top 17 candidate hub genes.Based on the differential expression analysis of these 17 candidate hub genes in the validation datasets,AEBP1 and COL8A2 were ultimately selected as the common key genes for both diseases,both of which showed a significant upregulation trend in the disease groups(all P<0.05).The value of area under the curve(AUC)for AEBP1 and COL8A2 in the OA and sarcopenia datasets were all greater than 0.7,indicating that both genes have potential value in predicting OA and sarcopenia.Real-time RT-PCR results showed that the mRNA expression levels of AEBP1 and COL8A2 were significantly upregulated in the disease groups(all P<0.05),consistent with the results observed in the bioinformatics analysis.GSEA revealed that AEBP1 and COL8A2 were closely related to extracellular matrix-receptor interaction,ribosome,and oxidative phosphorylation in OA and sarcopenia.Conclusion:AEBP1 and COL8A2 have the potential to serve as common biomarkers for OA and sarcopenia.The extracellular matrix-receptor interaction pathway may represent a potential target for the prevention and treatment of both OA and sarcopenia.展开更多
Objective:Keratoconus(KC)is a progressive corneal ectasia disorder,arising from a myriad of causes including genetic predispositions,environmental factors,biomechanical influences,and inflammatory reactions.This study...Objective:Keratoconus(KC)is a progressive corneal ectasia disorder,arising from a myriad of causes including genetic predispositions,environmental factors,biomechanical influences,and inflammatory reactions.This study aims to identify potential pathogenetic gene mutations in patients with sporadic KC in the Han Chinese population.Methods:Twenty-five patients with primary KC as well as 50 unrelated population matched healthy controls,were included in this study to identify potential pathogenic gene mutations among sporadic KC patients in the Han Chinese population.Sanger sequencing and whole-exome sequencing(WES)were used to analyze mutations in the zinc finger protein 469(ZNF469)gene.Bioinformatics analysis was conducted to explore the potential role of ZNF469 in KC pathogenesis.Results:Five novel heterozygous missense variants were identified in KC patients.Among them,2 compound heterozygous variants,c.8986G>C(p.E2996Q)with c.11765A>C(p.D3922A),and c.4423C>G(p.L1475V)with c.10633G>A(p.G3545R),were determined to be possible pathogenic factors for KC.Conclusion:Mutations in the ZNF469 gene may contribute to the development of KC in the Han Chinese population.These mutation sites may provide valuable information for future genetic screening of KC patients and their families.展开更多
Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were ra...Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were randomized equally into control group and heat stress group.After exposure to 32℃for 2 weeks in the latter group,the rats were examined for histopathological changes and Bmal1 expression in the thoracic aorta using HE staining and immunohistochemistry.In the cell experiments,cultured rat thoracic aortic endothelial cells(RTAECs)were incubated at 40℃for 12 h with or without prior transfection with a Bmal1-specific small interfering RNA(si-Bmal1)or a negative sequence.In both rat thoracic aorta and RTAECs,the expressions of Bmal1,the cell cycle proteins CDK1,CDK4,CDK6,and cyclin B1,and apoptosis-related proteins Bax and Bcl-2 were detected using Western blotting.TUNEL staining was used to detect cell apoptosis in rat thoracic aorta,and the changes in cell cycle distribution and apoptosis in RTAECs were analyzed with flow cytometry.Results Compared with the control rats,the rats exposed to heat stress showed significantly increased blood pressures and lowered heart rate with elastic fiber disruption and increased expressions of Bmal1,cyclin B1 and CDK1 in the thoracic aorta(P<0.05).In cultured RTAECs,heat stress caused significant increase of Bmal1,cyclin B1 and CDK1 protein expression levels,which were obviously lowered in cells with prior si-Bmal1 transfection.Bmal1 knockdown also inhibited heat stress-induced increase of apoptosis in RTAECs as evidenced by decreased expression of Bax and increased expression of Bcl-2.Conclusion Heat stress upregulates Bmal1 expression and causes alterations in expressions of cyclins to trigger apoptosis of rat thoracic aorta endothelial cells,which can be partly alleviated by suppressing Bmal1 expression.展开更多
Objective:Postpartum depression(PPD)is a common and serious mental disorder after childbirth,imposing a heavy burden on mothers,infants,and families.Abnormalities in the tryptophan-kynurenine(TRP-KYN)metabolic pathway...Objective:Postpartum depression(PPD)is a common and serious mental disorder after childbirth,imposing a heavy burden on mothers,infants,and families.Abnormalities in the tryptophan-kynurenine(TRP-KYN)metabolic pathway are considered to be involved in its pathogenesis,but the role of quinolinic acid phosphoribosyltransferase(QPRT),a key downstream enzyme in this pathway,remains unclear.This study aims to explore the association between PPD in women undergoing cesarean section and QPRT gene polymorphisms,as well as other risk factors for PPD.Methods:A candidate gene association study design was adopted.From January 2024 to June 2025,full-term singleton pregnant women scheduled to undergo elective cesarean section under spinal anesthesia were recruited at the Third Xiangya Hospital of Central South University and Hunan Provincial Maternal and Child Health Hospital.At 42 days postpartum,postpartum depression was assessed using the Edinburgh Postnatal Depression Scale(EPDS).Peripheral blood samples were collected and genomic DNA was extracted.Four QPRT single nucleotide polymorphism loci(rs1134700,rs2303255,rs9922666,and rs9933310)were selected for genotyping to analyze the association between these loci and PPD.Bioinformatics analysis and dual-luciferase reporter gene assays were performed to investigate the possible mechanism by which significant loci influence disease occurrence.Results:A total of 362 women were ultimately included in the analysis,among whom 29 were diagnosed with PPD,with an incidence of 8.01%.Analysis of general data showed that comorbid hypertension or thyroid disease,inconsistency between neonatal sex and expectation,prenatal depression,prenatal self-harm ideation,domestic violence,poor marital and mother-in-law/daughter-in-law relationships,stressful life events,dissatisfaction with current life status,poor mood during pregnancy,and high stress during pregnancy were all risk factors for PPD in women undergoing cesarean section(all P<0.05).Genetic association analysis revealed that the QPRT rs9933310 A>G polymorphism was associated with PPD.Women carrying the rs9933310 GG or AG genotype had a 2.92-fold higher risk of PPD compared with women with the AA genotype(OR=2.92,95%CI 1.18 to 6.99).Expression quantitative trait loci(eQTL)analysis suggested that the G allele at this locus was associated with downregulation of QPRT expression(AA>AG>GG).Multi-database queries indicated that the rs9933310 locus may have promoter and/or enhancer activity.In addition,JASPAR database prediction and experimental validation showed that the mutant(G)allele at the QPRT rs9933310 locus was more likely than the wild-type(A)allele to weaken promoter-enhancer activity at this locus,and resulted in loss of transcription factors Gata1,GATA2,GATA3,Gata4,Sox17,Sox2,Sox3,Sox6,and SRY,thereby regulating QPRT expression.Conclusion:Comorbid hypertension or thyroid disease,inconsistency between neonatal sex and expectation,prenatal depression,prenatal self-harm ideation,domestic violence,poor marital and mother-in-law/daughter-in-law relationships,stressful life events,dissatisfaction with current life status,poor mood during pregnancy,high stress during pregnancy,and mutation at the QPRT rs9933310 locus are all risk factors for PPD.The QPRT rs9933310 G allele is an independent risk factor for PPD in women undergoing cesarean section,and its pathogenic mechanism may involve downregulation of QPRT expression and disruption of TRP-KYN pathway homeostasis.QPRT has a potential role in the pathogenesis of PPD and may become a novel antidepressant target acting on the TRPKYN pathway.展开更多
Background Verticillium dahliae,a soil-borne fungi,can cause Verticillium wilt,and seriously diminish the yield and quality of cotton.However,the pathogenic mechanism of V.dahliae is complex and not clearly understood...Background Verticillium dahliae,a soil-borne fungi,can cause Verticillium wilt,and seriously diminish the yield and quality of cotton.However,the pathogenic mechanism of V.dahliae is complex and not clearly understood at the moment.This study aimed to identify the high-affinity nicotinic acid transporter genes in V.dahliae.The gene expression profiles in V.dahliae following sensing of root exudates from susceptible and resistant cotton varieties were analyzed.The function of VdNAT1 in the pathogenic process of V.dahliae was studied using the tobacco rattle virus(TRV)-based host-induced gene silencing(HIGS)technique.Results Eight high-affinity nicotinic acid transporter genes were identified from V.dahliae through the bioinformatics method.Each protein contains a conserved major facilitator superfamily(MFS)domain,which belongs to the MFS superfamily.Evolutionary relationship analysis revealed that all 8 genes belong to the anion:cation symporter(ACS)subfamily.All proteins have transmembrane domains,ranging from 7 to 12.The expression levels of most VdNAT genes were significantly increased after induction by root exudates from susceptible cotton varieties.Silencing VdNAT1 gene by HIGS significantly inhibited the accumulation of fungal biomass in cotton plants,and alleviated the disease symptoms of cotton.Conclusions Eight VdNAT genes were identified from V.dahliae,and most VdNAT genes was up-regulated after induced by root exudates from susceptible cotton variety.In addition,VdNAT1 is required for the pathogenicity of V.dahliae.Overall,these findings will facilitate the pathogenic molecular mechanism of V.dahliae and provide candidate genes.展开更多
Objective:Metabolic dysfunction-associated steatohepatitis(MASH),a progressive subtype of metabolic dysfunction-associated steatotic liver disease(MASLD),is characterized by hepatic steatosis,lobular inflammation,and ...Objective:Metabolic dysfunction-associated steatohepatitis(MASH),a progressive subtype of metabolic dysfunction-associated steatotic liver disease(MASLD),is characterized by hepatic steatosis,lobular inflammation,and hepatocyte ballooning,and may further progress to liver fibrosis and cirrhosis.Lectin-like oxidized low-density lipoprotein receptor-1(LOX-1),a member of the scavenger receptor family,recognizes and binds oxidized low-density lipoprotein.This study aims to investigate the role of LOX-1 in MASH progression.Methods:LOX-1 expression in MASLD mouse liver was analyzed using Gene Expression Omnibus(GEO)datasets.Immunofluorescence staining was performed to detect LOX-1 and alpha-smooth muscle actin(α-SMA)levels and co-localization in fibrotic liver tissues and LX-2 cells.LOX-1 knockout(Lox-1^(−/−))mice were generated using CRISPR/caspase-9(Cas9)and genotyped by PCR and Sanger sequencing.Wild-type(WT)and Lox-1^(−/−)mice were randomized into control and Western diet model groups.Serum and liver samples were collected for alanine aminotransferase(ALT)and aspartate aminotransferase(AST)measurement by biochemical kits,liver structure evaluation by hematoxylin and eosin(HE)staining,collagen deposition by Masson staining,lipid accumulation by Oil Red O staining,and fibrotic marker gene expression by real-time quantitative PCR(RT-qPCR).Network pharmacology and search tool for the retrieval of interacting genes/proteins(STRING)-based protein-protein interaction(PPI)with Gene Ontology(GO)enrichment were used to predict downstream targets and pathways.Results:The results from the GEO datasets GSE30552 and GSE40041 indicated LOX-1 mRNA was upregulated in high fat diet(HFD)and bile duct ligation(BDL)mouse models(both P<0.001).LOX-1 and α-SMA levels were elevated in fibrotic liver tissues.Lox-1^(−/−)mice were successfully established.Biochemical tests showed that serum AST and ALT levels were significantly elevated in WT mice fed a Western diet(both P<0.001),and these levels decreased after LOX-1 knockout(both P<0.05).HE staining revealed that WT mice on the Western diet exhibited marked hepatocellular ballooning degeneration,steatosis,inflammatory cell infiltration,and periportal fibroplasia,which were significantly ameliorated by LOX-1 knockout.Masson staining demonstrated increased blue-stained collagen fibers in the liver tissues of WT mice fed the Western diet compared with controldiet mice,and LOX-1 knockout inhibited collagen fiber deposition(all P<0.05).RT‑qPCR results showed that hepatic mRNA levels of Acta2,Col1a1,and Timp1 were significantly increased in Western diet-fed mice,and LOX-1 knockout reduced the expression of these fibrogenic marker genes.Oil Red O staining indicated that hepatocytes in WT mice fed the Western diet were notably enlarged,displayed macrovesicular steatosis,and exhibited diffusely distributed red lipid droplets,whereas LOX-1 knockout alleviated hepatic lipid accumulation(both P<0.001).RT‑qPCR results further demonstrated that knockdown of LOX-1 reduced Acta2,Col1a1,and Timp1 mRNA levels in LX‑2 cells(all P<0.05).Immunofluorescence analysis revealed co‑localization of LOX-1 and α‑SMA in LX‑2 cells,and LOX-1 silencing suppressed α‑SMA expression.Network pharmacology suggested LOX-1 may promote MASH via lipid and cholesterol metabolism networks.Conclusion:LOX-1 gene knockout ameliorates Western diet-induced MASH in mice and may serve as a potential therapeutic target.展开更多
文摘A recently published study(Xin et al.,Prog Biochem Biophys,2026,53(2):431-441.DOI:10.3724/j.pibb.2025.0508)addresses the therapeutic challenges of pancreatic ductal adenocarcinoma(PDAC)by innovatively developing an orally administered nanogene delivery system.Designed to achieve in situ,efficient delivery of chimeric antigen receptor(CAR)genes to tumor sites,this approach offers a novel strategy for CAR-macrophage(CAR-M)based immunotherapy.Its key highlights are as follows.
基金Supported by the National Key R&D Program(2022YFF1300500)the Spring Goose Support Program(CYQN24018)the National Natural Science Foundation of China(32572123)。
文摘Dianthus spiculifolius Schur,as an emerging ornamental plant,has extensive applications and economic values.In this study,the DsCBL4 gene was successfully cloned,and its tissue-specific expression,expression patterns under various abiotic stresses,subcellular localization,and bioinformatics analysis of the encoded amino acid sequence were conducted.The results showed that the coding region of the DsCBL4 gene was 675 bp long,encoding 224 amino acids.It had high homology with the amino acids encoded by Amaranthus tricolor,Chenopodium quinoa and Spinacia oleracea.The predicted relative molecular mass of DsCBL4 was 25.61 ku,with an isoelectric point of 4.58,and it had phosphorylation sites,belonging to an unstable hydrophilic protein.Its secondary structure includedα-helices,irregular coils and extended chains.The tertiary structure prediction revealed that DsCBL4 had four EFhand calcium-binding domains necessary for Ca2+binding in plant calmodulin-like proteins and the FPSF motif for calcineurin B-like protein(CBL)-interacting protein kinase(CIPK)activation.The expression level of the DsCBL4 gene showed tissue specificity,with the highest expression in roots.It was induced by drought,low temperature,combined drought and low temperature,salt stress,nitrogen stress,phosphorus stress,calcium ion stress,high temperature stress,and abscisic acid(ABA)stress.Both transient infection in Nicotiana tabacum L.and stable expression in transgenic Arabidopsis thaliana showed that the DsCBL4 protein was localized to the cell membrane.These results suggested that DsCBL4 might be involved in the abiotic stress response of Dianthus spiculifolius through the calcium signaling pathway,providing a theoretical basis for understanding its molecular mechanism.This study provided an important reference for further exploring the role of the DsCBLs gene family in plant stress resistance.
基金supported by the National Natural Science Foundation of China(82060418).
文摘Objective:Osteoarthritis(OA)and sarcopenia are significant health concerns in the elderly,substantially impacting their daily activities and quality of life.However,the relationship between them remains poorly understood.This study aims to uncover common biomarkers and pathways associated with both OA and sarcopenia.Methods:Gene expression profiles related to OA and sarcopenia were retrieved from the Gene Expression Omnibus(GEO)database.Differentially expressed genes(DEGs)between disease and control groups were identified using R software.Common DEGs were extracted via Venn diagram analysis.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were conducted to identify biological processes and pathways associated with shared DEGs.Protein-protein interaction(PPI)networks were constructed,and candidate hub genes were ranked using the maximal clique centrality(MCC)algorithm.Further validation of hub gene expression was performed using 2 independent datasets.Receiver operating characteristic(ROC)curve analysis was used to evaluate the predictive value of key genes for OA and sarcopenia.Mouse models of OA and sarcopenia were established.Hematoxylin-eosin and Safranin O/Fast Green staining were used to validate the OA model.The sarcopenia model was validated via rotarod testing and quadriceps muscle mass measurement.Real-time reverse transcription PCR(real-time RT-PCR)was employed to assess the mRNA expression levels of candidate key genes in both models.Gene set enrichment analysis(GSEA)was conducted to identify pathways associated with the selected shared key genes in both diseases.Results:A total of 89 common DEGs were identified in the gene expression profiles of OA and sarcopenia,including 76 upregulated and 13 downregulated genes.These 89 DEGs were significantly enriched in protein digestion and absorption,the PI3K-Akt signaling pathway,and extracellular matrix-receptor interaction.PPI network analysis and MCC algorithm analysis of the 89 common DEGs identified the top 17 candidate hub genes.Based on the differential expression analysis of these 17 candidate hub genes in the validation datasets,AEBP1 and COL8A2 were ultimately selected as the common key genes for both diseases,both of which showed a significant upregulation trend in the disease groups(all P<0.05).The value of area under the curve(AUC)for AEBP1 and COL8A2 in the OA and sarcopenia datasets were all greater than 0.7,indicating that both genes have potential value in predicting OA and sarcopenia.Real-time RT-PCR results showed that the mRNA expression levels of AEBP1 and COL8A2 were significantly upregulated in the disease groups(all P<0.05),consistent with the results observed in the bioinformatics analysis.GSEA revealed that AEBP1 and COL8A2 were closely related to extracellular matrix-receptor interaction,ribosome,and oxidative phosphorylation in OA and sarcopenia.Conclusion:AEBP1 and COL8A2 have the potential to serve as common biomarkers for OA and sarcopenia.The extracellular matrix-receptor interaction pathway may represent a potential target for the prevention and treatment of both OA and sarcopenia.
基金supported by the National Natural Science Foundation(82271057)the Natural Science Foundation of Hunan Province(2023JJ30818),China。
文摘Objective:Keratoconus(KC)is a progressive corneal ectasia disorder,arising from a myriad of causes including genetic predispositions,environmental factors,biomechanical influences,and inflammatory reactions.This study aims to identify potential pathogenetic gene mutations in patients with sporadic KC in the Han Chinese population.Methods:Twenty-five patients with primary KC as well as 50 unrelated population matched healthy controls,were included in this study to identify potential pathogenic gene mutations among sporadic KC patients in the Han Chinese population.Sanger sequencing and whole-exome sequencing(WES)were used to analyze mutations in the zinc finger protein 469(ZNF469)gene.Bioinformatics analysis was conducted to explore the potential role of ZNF469 in KC pathogenesis.Results:Five novel heterozygous missense variants were identified in KC patients.Among them,2 compound heterozygous variants,c.8986G>C(p.E2996Q)with c.11765A>C(p.D3922A),and c.4423C>G(p.L1475V)with c.10633G>A(p.G3545R),were determined to be possible pathogenic factors for KC.Conclusion:Mutations in the ZNF469 gene may contribute to the development of KC in the Han Chinese population.These mutation sites may provide valuable information for future genetic screening of KC patients and their families.
文摘Objective To investigate the structural changes of rat thoracic aorta and changes in expression levels of Bmal1 and cyclins in thoracic aorta endothelial cells following heat stress.Methods Twenty male SD rats were randomized equally into control group and heat stress group.After exposure to 32℃for 2 weeks in the latter group,the rats were examined for histopathological changes and Bmal1 expression in the thoracic aorta using HE staining and immunohistochemistry.In the cell experiments,cultured rat thoracic aortic endothelial cells(RTAECs)were incubated at 40℃for 12 h with or without prior transfection with a Bmal1-specific small interfering RNA(si-Bmal1)or a negative sequence.In both rat thoracic aorta and RTAECs,the expressions of Bmal1,the cell cycle proteins CDK1,CDK4,CDK6,and cyclin B1,and apoptosis-related proteins Bax and Bcl-2 were detected using Western blotting.TUNEL staining was used to detect cell apoptosis in rat thoracic aorta,and the changes in cell cycle distribution and apoptosis in RTAECs were analyzed with flow cytometry.Results Compared with the control rats,the rats exposed to heat stress showed significantly increased blood pressures and lowered heart rate with elastic fiber disruption and increased expressions of Bmal1,cyclin B1 and CDK1 in the thoracic aorta(P<0.05).In cultured RTAECs,heat stress caused significant increase of Bmal1,cyclin B1 and CDK1 protein expression levels,which were obviously lowered in cells with prior si-Bmal1 transfection.Bmal1 knockdown also inhibited heat stress-induced increase of apoptosis in RTAECs as evidenced by decreased expression of Bax and increased expression of Bcl-2.Conclusion Heat stress upregulates Bmal1 expression and causes alterations in expressions of cyclins to trigger apoptosis of rat thoracic aorta endothelial cells,which can be partly alleviated by suppressing Bmal1 expression.
基金supported by the Natural Science Foundation of Hunan Province,China(2018JJ2598).
文摘Objective:Postpartum depression(PPD)is a common and serious mental disorder after childbirth,imposing a heavy burden on mothers,infants,and families.Abnormalities in the tryptophan-kynurenine(TRP-KYN)metabolic pathway are considered to be involved in its pathogenesis,but the role of quinolinic acid phosphoribosyltransferase(QPRT),a key downstream enzyme in this pathway,remains unclear.This study aims to explore the association between PPD in women undergoing cesarean section and QPRT gene polymorphisms,as well as other risk factors for PPD.Methods:A candidate gene association study design was adopted.From January 2024 to June 2025,full-term singleton pregnant women scheduled to undergo elective cesarean section under spinal anesthesia were recruited at the Third Xiangya Hospital of Central South University and Hunan Provincial Maternal and Child Health Hospital.At 42 days postpartum,postpartum depression was assessed using the Edinburgh Postnatal Depression Scale(EPDS).Peripheral blood samples were collected and genomic DNA was extracted.Four QPRT single nucleotide polymorphism loci(rs1134700,rs2303255,rs9922666,and rs9933310)were selected for genotyping to analyze the association between these loci and PPD.Bioinformatics analysis and dual-luciferase reporter gene assays were performed to investigate the possible mechanism by which significant loci influence disease occurrence.Results:A total of 362 women were ultimately included in the analysis,among whom 29 were diagnosed with PPD,with an incidence of 8.01%.Analysis of general data showed that comorbid hypertension or thyroid disease,inconsistency between neonatal sex and expectation,prenatal depression,prenatal self-harm ideation,domestic violence,poor marital and mother-in-law/daughter-in-law relationships,stressful life events,dissatisfaction with current life status,poor mood during pregnancy,and high stress during pregnancy were all risk factors for PPD in women undergoing cesarean section(all P<0.05).Genetic association analysis revealed that the QPRT rs9933310 A>G polymorphism was associated with PPD.Women carrying the rs9933310 GG or AG genotype had a 2.92-fold higher risk of PPD compared with women with the AA genotype(OR=2.92,95%CI 1.18 to 6.99).Expression quantitative trait loci(eQTL)analysis suggested that the G allele at this locus was associated with downregulation of QPRT expression(AA>AG>GG).Multi-database queries indicated that the rs9933310 locus may have promoter and/or enhancer activity.In addition,JASPAR database prediction and experimental validation showed that the mutant(G)allele at the QPRT rs9933310 locus was more likely than the wild-type(A)allele to weaken promoter-enhancer activity at this locus,and resulted in loss of transcription factors Gata1,GATA2,GATA3,Gata4,Sox17,Sox2,Sox3,Sox6,and SRY,thereby regulating QPRT expression.Conclusion:Comorbid hypertension or thyroid disease,inconsistency between neonatal sex and expectation,prenatal depression,prenatal self-harm ideation,domestic violence,poor marital and mother-in-law/daughter-in-law relationships,stressful life events,dissatisfaction with current life status,poor mood during pregnancy,high stress during pregnancy,and mutation at the QPRT rs9933310 locus are all risk factors for PPD.The QPRT rs9933310 G allele is an independent risk factor for PPD in women undergoing cesarean section,and its pathogenic mechanism may involve downregulation of QPRT expression and disruption of TRP-KYN pathway homeostasis.QPRT has a potential role in the pathogenesis of PPD and may become a novel antidepressant target acting on the TRPKYN pathway.
基金supported by National Natural Science Foundation of China(No.32160615).
文摘Background Verticillium dahliae,a soil-borne fungi,can cause Verticillium wilt,and seriously diminish the yield and quality of cotton.However,the pathogenic mechanism of V.dahliae is complex and not clearly understood at the moment.This study aimed to identify the high-affinity nicotinic acid transporter genes in V.dahliae.The gene expression profiles in V.dahliae following sensing of root exudates from susceptible and resistant cotton varieties were analyzed.The function of VdNAT1 in the pathogenic process of V.dahliae was studied using the tobacco rattle virus(TRV)-based host-induced gene silencing(HIGS)technique.Results Eight high-affinity nicotinic acid transporter genes were identified from V.dahliae through the bioinformatics method.Each protein contains a conserved major facilitator superfamily(MFS)domain,which belongs to the MFS superfamily.Evolutionary relationship analysis revealed that all 8 genes belong to the anion:cation symporter(ACS)subfamily.All proteins have transmembrane domains,ranging from 7 to 12.The expression levels of most VdNAT genes were significantly increased after induction by root exudates from susceptible cotton varieties.Silencing VdNAT1 gene by HIGS significantly inhibited the accumulation of fungal biomass in cotton plants,and alleviated the disease symptoms of cotton.Conclusions Eight VdNAT genes were identified from V.dahliae,and most VdNAT genes was up-regulated after induced by root exudates from susceptible cotton variety.In addition,VdNAT1 is required for the pathogenicity of V.dahliae.Overall,these findings will facilitate the pathogenic molecular mechanism of V.dahliae and provide candidate genes.
基金supported by the Natural Science Foundation of Hunan Province,China(211142095031)。
文摘Objective:Metabolic dysfunction-associated steatohepatitis(MASH),a progressive subtype of metabolic dysfunction-associated steatotic liver disease(MASLD),is characterized by hepatic steatosis,lobular inflammation,and hepatocyte ballooning,and may further progress to liver fibrosis and cirrhosis.Lectin-like oxidized low-density lipoprotein receptor-1(LOX-1),a member of the scavenger receptor family,recognizes and binds oxidized low-density lipoprotein.This study aims to investigate the role of LOX-1 in MASH progression.Methods:LOX-1 expression in MASLD mouse liver was analyzed using Gene Expression Omnibus(GEO)datasets.Immunofluorescence staining was performed to detect LOX-1 and alpha-smooth muscle actin(α-SMA)levels and co-localization in fibrotic liver tissues and LX-2 cells.LOX-1 knockout(Lox-1^(−/−))mice were generated using CRISPR/caspase-9(Cas9)and genotyped by PCR and Sanger sequencing.Wild-type(WT)and Lox-1^(−/−)mice were randomized into control and Western diet model groups.Serum and liver samples were collected for alanine aminotransferase(ALT)and aspartate aminotransferase(AST)measurement by biochemical kits,liver structure evaluation by hematoxylin and eosin(HE)staining,collagen deposition by Masson staining,lipid accumulation by Oil Red O staining,and fibrotic marker gene expression by real-time quantitative PCR(RT-qPCR).Network pharmacology and search tool for the retrieval of interacting genes/proteins(STRING)-based protein-protein interaction(PPI)with Gene Ontology(GO)enrichment were used to predict downstream targets and pathways.Results:The results from the GEO datasets GSE30552 and GSE40041 indicated LOX-1 mRNA was upregulated in high fat diet(HFD)and bile duct ligation(BDL)mouse models(both P<0.001).LOX-1 and α-SMA levels were elevated in fibrotic liver tissues.Lox-1^(−/−)mice were successfully established.Biochemical tests showed that serum AST and ALT levels were significantly elevated in WT mice fed a Western diet(both P<0.001),and these levels decreased after LOX-1 knockout(both P<0.05).HE staining revealed that WT mice on the Western diet exhibited marked hepatocellular ballooning degeneration,steatosis,inflammatory cell infiltration,and periportal fibroplasia,which were significantly ameliorated by LOX-1 knockout.Masson staining demonstrated increased blue-stained collagen fibers in the liver tissues of WT mice fed the Western diet compared with controldiet mice,and LOX-1 knockout inhibited collagen fiber deposition(all P<0.05).RT‑qPCR results showed that hepatic mRNA levels of Acta2,Col1a1,and Timp1 were significantly increased in Western diet-fed mice,and LOX-1 knockout reduced the expression of these fibrogenic marker genes.Oil Red O staining indicated that hepatocytes in WT mice fed the Western diet were notably enlarged,displayed macrovesicular steatosis,and exhibited diffusely distributed red lipid droplets,whereas LOX-1 knockout alleviated hepatic lipid accumulation(both P<0.001).RT‑qPCR results further demonstrated that knockdown of LOX-1 reduced Acta2,Col1a1,and Timp1 mRNA levels in LX‑2 cells(all P<0.05).Immunofluorescence analysis revealed co‑localization of LOX-1 and α‑SMA in LX‑2 cells,and LOX-1 silencing suppressed α‑SMA expression.Network pharmacology suggested LOX-1 may promote MASH via lipid and cholesterol metabolism networks.Conclusion:LOX-1 gene knockout ameliorates Western diet-induced MASH in mice and may serve as a potential therapeutic target.
