Objective:To investigate the association of circadian syndrome(CircS)with the risks of 10 incident chronic diseases and multimorbidity(≥2 diseases).Methods:Based on the baseline data from the 2011 China Health and Retirement Longitudinal Study(CHARLS),8,093 participants were enrolled(CircS group:n=3,091;non-CircS group:n=5,002).Cox proportional hazards models were used to assess the association of CircS with the risks of incident chronic diseases and multimorbidity.Additionally,dose-response analyses,stratified analyses by follow-up duration(≤4 years vs＞4 years),and analyses of dynamic changes in CircS status were performed.Results:CircS was significantly associated with the increased risks of incident stroke(HR=2.07),memory-related diseases(HR=1.62),liver diseases(HR=1.38),heart diseases(HR=1.27),kidney diseases(HR=1.28)and digestive system diseases(HR=1.23).Dose-response analyses revealed that the participants with CircS scores≥6 points had a 170%increase in the risk of incident stroke(HR=2.70,95%CI:2.12-3.44),and a 66%increase in the risk of incident heart diseases(HR=1.66,95%CI:1.35-2.04).The risk of incident stroke was significantly increased during the short-term follow-up(≤4 years),whereas the risks of incident kidney diseases,digestive sys-tem diseases,and liver diseases emerged predominantly after the long-term follow-up(＞4 years).Regarding mul-timorbidity,CircS was significantly associated with higher risks of developing≥2(HR=1.30),≥3(HR=1.42),≥4(HR=1.53)and≥5(HR=1.49)diseases in the participants,and the risks of incident multimorbidity(≥5 diseases)were most pronounced in those with CircS scores≥6 points(HR=2.74).The analyses of dynamic changes in CircS status revealed that persistent CircS conferred the highest risk of incident diseases(stroke:HR=3.04).Nota-bly,residual risks of incident stroke,heart diseases and liver diseases remained even after CircS remission.Con-clusion:CircS is significantly associated with the risks of incident multiple chronic diseases and multimorbidity.This association exhibits dose-response gradients and time-varying patterns.The risks of incident diseases remain even after CircS remission,suggesting that CircS may serve as a potential indicator for the risk assessment of chronic diseases.
Objective To identify common hepatitis B virus (HBV) integration target genes and fragments in the liver tissues from patients with HBV-related hepatocellular carcinoma (HCC), thereby providing a theoretical basis for optimizing the early screening strategies of liver cancer. Methods Liquid-phase hybridization capture sequencing with HBV probes was applied to detect HBV integration in the cancerous and adjacent non-cancerous tissues from 26 patients with HBV-related HCC, and the patterns of HBV integration as well as their association with the pathogenesis of early-stage HCC were explored. Targeted amplification and Sanger sequencing were performed to validate HBV integration within the FN1 gene locus in a subset of samples from adjacent non-cancerous tissues. Results The number of HBV integration breakpoints detected in the cancerous tissues was significantly lower than that in the adjacent non-cancerous tissues. In early-stage HCC cases, HBV integration was predominantly observed in the adjacent non-cancerous tissues (P＜0.05). TERT was the most common target gene of HBV integration in the cancerous tissues, whereas FN1 was the predominant target gene of HBV integration in the adjacent non-cancerous tissues. HBV integrated fragments were primarily clustered in the 300–500 bp and 1300–1900 bp regions of the HBV genome, and the regions encompassed the entire HBX gene, Precore/core gene, and partial S genes. Finally, the precise integration of HBV within the FN1 gene locus was validated using polymerase chain reaction (PCR) and Sanger sequencing. The integration breakpoints of target genes in the cancerous tissues were mostly located in exons, whereas those in the adjacent non-cancerous tissues were mostly located in introns. Conclusion HBV integration is significantly enriched in the adjacent non-cancerous tissues of early-stage HCC, with FN1 being the most common target gene. Since the integrated HBV fragments can be released into the bloodstream, this study provides a preliminary theoretical basis for the subsequent development of non-invasive early screening techniques to detect HBV integration based on plasma cell-free DNA (cfDNA).
Objective To investigate the association between ambient temperature and the lethality of suicide methods, and to evaluate the potential effect modification by age, sex and season. Methods A retrospective analysis was conducted on suicide-related emergency records from the 120 dispatch system of Wuhan Emergency Medical Center between June 2023 and May 2025, which were matched with concurrent meteorological data. The suicide methods were classified as high-lethality and low-lethality based on case fatality rate. Binary logistic regression was used to assess the association between daily mean ambient temperature and the high-lethality suicide methods, with adjustment for age and sex. Subgroup analyses were stratified by age, sex and season. Multiple comparisons were corrected using false discovery rate (FDR), and the robustness was evaluated through 10 sensitivity analyses. Results A total of 1, 160 individuals were included in this study, among whom 287 individuals (24.7%) involved the high-lethality suicide methods. Each 1℃ increase in daily mean ambient temperature was associated with a 2.2% reduction in the odds of selecting a high-lethality suicide method (OR=0.978, 95% CI: 0.964–0.993, P=0.004). After FDR correction, the temperature effects in the 30–59 age group and in autumn reached statistical significance; however, likelihood ratio tests for interactions revealed no significant effect modification (P > 0.15). In the method-specific analyses, sharp instrument injuries (OR=1.021) and falls from height (OR=0.969) remained significant after FDR correction, and the direction of effects across methods was consistent with the lethality classification framework. The results were consistent across the 10 sensitivity analyses, with ORs remaining stable in the range of 0.976–0.985. Conclusion Ambient temperature is negatively associated with the lethality of suicide methods, and rising temperatures are accompanied by a decline in the proportion of the high-lethality suicide methods. These results remain robust across multiple sensitivity analyses.
Objective:To investigate the impact of diabetes mellitus(DM)on the risk of developing urinary tract tumors.Methods:Two-sample Mendelian randomization(MR)analyses were conducted using summary statis-tics from the IEU OpenGWAS and FinnGen databases.Phenotypic validation was conducted using cross-sectional population analysis,and secondary MR analysis was performed to enhance the strength of evidence.Results:Preliminary MR analysis revealed a weak association between type 2 diabetes mellitus(T2DM)and bladder cancer(OR=1.001,95%CI:1.000-1.001,P=0.010).DM(OR=1.016,95%CI:1.001-1.032,P=0.037)and T2DM(OR=1.002,95%CI:1.001-1.002,P=0.021)were confirmed as risk factors for prostate cancer(PCa),with the findings supported by secondary MR,and were validated by cross-sectional analysis.Subgroup analysis showed that T2DM increased the risk of PCa in situ(OR=2.114,95%CI:1.062-4.240,P=0.032).Conclusion:T2DM increases the risk of PCa,especially for carcinoma in situ.
β-Thalassemia is an inherited hemolytic anemia caused by defects in the β-globin gene.β-Thalasse-mia major results from homozygous or compound heterozygous β⁰ or β⁺ mutations.The 8p11 myeloproli-ferative syndrome(EMS)is a myeloproliferative neoplasm associated with gene translocation of fibroblast growth factor receptor 1(FGFR1)on the short arm of chromosome 8(8p11).BCR-FGFR1 represents a specific fusion gene sub-type of this syndrome,which tends to progress to leukemia with a poor prognosis.Allogeneic hematopoietic stem cell transplantation(allo-HSCT)is currently the only therapeutic approach expected to achieve long-term remis-sion.This article reports,for the first time in the world,a case of pediatric EMS arising in the context of β-thalas-semia major.The patient developed EMS driven by the BCR-FGFR1 fusion gene.Conventional cytogenetic analysis revealed,for the first time,an atypical translocation,t(8;21)(p11.2;q11.2),accompanied by a secondary deletion del(22)(q13).Although BCR-FGFR1 fusions have been well documented,this precise molecular configu-ration and its associated cytogenetic background have not been described to date.Given the lack of established therapeutic experience in patients with concurrent β-thalassemia major and EMS,strategies were formulated in line with mainstream international regimens and current clinical guidelines.The patient received intensive chemo-therapy combined with early allo-HSCT,resulting in gradual clinical improvement.At 47 months post-transplantation,the patient remains in good general condition.The findings of this study expand the molecular spectrum of FGFR1-driven neoplasms and provide novel clinical insights into the management of thalassemia complicated with hematologic malignancies.Furthermore,we review potential mechanisms underlying the de-velopment of hematological malignancies in thalassemia major patients and underscore the pivotal role of high-resolution genomic profiling and allo-HSCT in achieving durable remission.
Objective:To develop a knowledge assessment questionnaire for childhood developmental disorders,to test its reliability and validity,and to explore its application value among caregivers of children with develop-mental disorders.Methods:Based on expert consensus and relevant guidelines,two rounds of expert consultation and a pre-test were conducted to form the questionnaire.Stratified sampling was employed to select six children's rehabilitation institutions across regions with varying levels of economic development in Hunan Province as the study sites,and a total of 252 primary caregivers of children with developmental disorders were recruited for the investigation.The knowledge assessment questionnaire and medical-seeking behavior questionnaire were used to collect data for further testing the reliability and validity of the developed questionnaire.By using the median grouping method,the caregivers were divided into a high-knowledge group and a low-knowledge group.The dif-ferences in medical treatment and rehabilitation behaviors were compared between the two groups,and the refer-ence cut-off value of the questionnaire was determined accordingly.Results:The finally developed knowledge questionnaire consisted of 4 dimensions and 18 items.The item-level content validity index(I-CVI)ranged from 0.93 to 1.00,and the scale-level content validity index(S-CVI)was 0.98.Confirmatory factor analysis results revealed that the model fitted well[comparative fitted index(CFI)=0.898,root mean square error of approxima-tion(RMSEA)=0.069].The overall Cronbach's α coefficient of the questionnaire was 0.798,and both McDonald's ω coefficients and Cronbach's α coefficient of each dimension were all greater than 0.7.Based on the median total scores of the questionnaire(15 points),caregivers in the low-knowledge group(scores≤15 points)were more likely to experience delayed diagnosis and treatment,and showed poorer compliance with rehabilitation treat-ment.Therefore,it was suggested that 15 points be taken as the reference cut-off value of the questionnaire.Care-givers with scores≤15 points were considered to have inadequate knowledge and require targeted health educa-tion.Conclusion:The knowledge assessment questionnaire for childhood developmental disorders has good reli-ability,validity and discrimination ability.It can be applied to scientific research and clinical health education practice related to the diagnosis,treatment,and rehabilitation of childhood developmental disorders.
Objective:To investigate the expression and function of Vasorin(Vasn)in the polarization of mouse monocyte-macrophage leukemia cells(RAW 264.7),providing a foundation for the mechanistic study of various inflammatory diseases.Methods:Macrophages were induced to undergo M1/M2 polarization using lipopolysac-charide(LPS)and interleukin(IL)-4 respectively,and the expression of Vasn was detected.A stable Vasn-overexpressing cell line was constructed using lentiviral technology.Experimental groups included a control(con-trol)group,a negative control(NC)group,and an overexpression(OE)group.Additionally,each of these groups was further subdivided into untreated,LPS-treated,and IL-4-treated subgroups.Expressions of polarization mark-ers and inflammatory factors were assessed using nitric oxide(NO)detection,reverse transcription-quantitative polymerase chain reaction(RT-qPCR),enzyme-linked immunosorbent assay(ELISA),and western blotting(WB).In addition,differentially expressed genes were screened through transcriptome sequencing,and the core signaling pathways were identified by GO functional and KEGG pathway enrichment analysis.Finally,the phos-phorylation levels of key proteins in the pathways were verified by WB experiments.Results:The expressions of Vasn mRNA and Vasn protein were significantly downregulated following LPS stimulation,and were signifi-cantly upregulated following IL-4 stimulation(all P＜0.05).Compared with those in the NC group,the Vasn mRNA and Vasn protein expression levels in the OE group were significantly elevated(all P＜0.05).After LPS stimulation,the cellular NO concentration,secretion levels and mRNA expressions of inflammatory factors in-cluding mouse tumor necrosis factor-alpha(TNF-α),IL-6 and IL-1β,as well as the M1 polarization marker pro-teins cluster of differentiation 86(CD86)and inducible nitric oxide synthase(iNOS),were significantly upregu-lated.In contrast,the secretion levels,marker gene expression,and protein expressions of M1 polarization factors in the OE group were significantly lower than those in the NC group(all P＜0.05).GO functional enrichment analysis and KEGG pathway enrichment were used to identify important signaling pathways including NF-κB.In the OE group,the phosphorylation level of p65,a key protein in the NF-κB pathway,was significantly down-regulated(P＜0.05),which was consistent with the sequencing results.After IL-4 stimulation,the levels of IL-10,the M2 polarization marker proteins arginase-1(ARG1)and mannose receptor(CD206),as well as the mRNA ex-pressions of the M2 polarization genes ARG1,chitinase 3-like protein 1(YM1),and IL-10 were significantly el-evated.Moreover,the secretion levels,marker gene expression,and protein expression of M2 polarization factors in the OE group were significantly higher than those in the NC group(all P＜0.05).GO functional enrichment analysis and KEGG pathway enrichment were uesd to identify important signaling pathways including JAK-STAT.In the OE group,the phosphorylation level of STAT6,a key protein in the JAK-STAT signaling pathway,was significantly enhanced(P＜0.05),which was consistent with the sequencing results.Conclusion:Vasn not only inhibits LPS-induced M1 polarization of macrophages by regulating the NF-κB pathway,alleviating the in-flammatory response,but also enhances IL-4-induced M2 polarization and anti-inflammatory responses by pro-moting STAT6 phosphorylation.
Objective:To construct a quantum dot-based electrochemical sensor amplified by terminal deoxy-nucleotidyl transferase(TdT)for the ultrasensitive detection of epithelial cell adhesion molecule(EpCAM)on the surface of tumor cells.Methods:A two-step hydrothermal method was adopted to synthesize DNA-functionalized quantum dot(QD-DNA).TdT was used to construct extended long polymeric aptamers.The two components were hybridized to obtain conjugated probes,and an electrochemical sensor was further fabricated.The reproducibility,anti-interference capability and specificity of the sensor were systematically investigated;meanwhile,the phenotypic changes of MCF-7 cells during drug treatment were monitored.Results:The QD-based electrochemical sensor amplified by TdT was successfully fabricated,with a detection limit as low as 80 cells,showing favorable detection sensitivity,specificity,accuracy and anti-interference capability.The sensor could effectively detect the expression levels of EpCAM on different cell surfaces and dynamically monitor the phenotypic changes of tumor cells during drug treatment.Conclusion:The QD-based electrochemical sensor am-plified by TdT can effectively distinguish tumor cells from normal cells and dynamically monitor the phenotypic changes of tumor cells during drug treatment,showing promising application potential in early tumor diagnosis and precise screening.
Objective:To assess the associations between different metabolic obesity phenotypes and the risks of hypertension,chronic kidney disease(CKD),and hyperuricemia among adults in Southern China.Methods:A to-tal of 8,972 participants were recruited using convenience sampling in this study.According to body mass index(BMI)and metabolic health status,all participants were divided into four groups:the metabolically healthy non-overweight/obesity(MHNO)group,the metabolically unhealthy non-overweight/obesity(MUNO)group,the metabolically healthy overweight/obesity(MHO)group,and the metabolically unhealthy overweight/obesity(MUO)group.Multivariable logistic regression models were applied to analyze the associations of different meta-bolic obesity phenotypes with hypertension,CKD,and hyperuricemia.Stratified analyses were further conducted by gender to explore the heterogeneity.Results:Compared with the MHNO group,the risks of hypertension in both the MUNO and MUO groups(MUNO group:OR=2.47,95%CI:2.09-2.91;MUO group:OR=3.08,95%CI:2.62-3.63)were significantly increased,with the MUO group showing the highest risk.Meanwhile,com-pared with the MHNO group,both the MUNO and MUO groups had a significantly increased risk of CKD(MUNO group:OR=2.81,95%CI:1.95-4.05;MUO group:OR=3.15,95%CI:2.20-4.51),as well as a signifi-cantly increased risk of elevated urinary albumin-to-creatinine ratio(uACR)(MUNO group:OR=1.90,95%CI:1.39-2.61;MUO group:OR=2.37,95%CI:1.72-3.26).The risk of hyperuricemia was increased in the MUNO,MHO and MUO groups,with the MUO group exhibiting the highest risk(OR=4.32,95%CI:3.47-5.37).Gender-stratified analysis revealed that these associations were consistent across both males and females.Conclusion:Metabolically unhealthy status is significantly associated with the risk of multiple cardiometabolic and renal dis-eases,and this association is further strengthened when combined with obesity.
Bronchial asthma is a heterogeneous disease characterized by chronic airway inflammation,reversible airflow limitation and airway remodeling.Airway remodeling is the key pathological basis for the progression of mild asthma to severe and refractory asthma.As an early core event and primary feature of airway remodeling,epithelial-mesenchymal transition(EMT)is initiated by direct damage to the airway epithelial barrier caused by harmful stimuli such as allergens,PM2.5 and tobacco smoke.It is coordinately regulated by inflammatory factors including TGF-β,the interleukin family,TNF-α and signaling pathways such as PI3K/AKT,Smad and Wnt/β-catenin,accompanied by downregulated expression of E-cadherin and elevated levels of mesenchymal markers including α-SMA and vimentin.Non-coding RNAs(miRNA,lncRNA)and key genes such as PER2 and GLCCI1 participate in the fine regulation of EMT at the transcriptional and epigenetic levels.At present,biologics,macro-lides,hormones,signaling pathway inhibitors and traditional Chinese medicine have shown potential in inhibiting EMT and improving airway remodeling.However,clinical translation still faces challenges including disturbance of physiological functions,phenotypic heterogeneity,unclear long-term safety and insufficient large-sample clini-cal evidence.Further elucidation of the regulatory network of EMT in asthma airway remodeling and exploration of precise,reversible and safe targeted intervention strategies are of great theoretical value and clinical signifi-cance for reversing airway remodeling and ameliorating the prognosis of severe and refractory asthma.
Objective:To explore the association between tumor necrosis factor(TNF)gene polymorphisms and cognitive aging in longevity areas of Guangxi.Methods:A total of 1,129 elderly Zhuang individuals(575 males,554 females),aged over 60 years,from the Hongshui River Basin and Hezhou region in Guangxi were included.They were divided into the longevous elderly(93.56±3.53 years),offspring of the longevous elderly(68.40±4.81 years),local elderly controls(69.30±5.50 years),and non-local elderly controls(68.93±5.61 years).The partici-pants'educational level,blood glucose,blood lipids,C-reactive protein(CRP)and other indicators were recorded.Cognitive status was assessed and defined by the mini-mental state examination(MMSE)in the study population.Genotyping of the TNF gene was performed using the SNPscanTM multiplex single nucleotide polymorphism(SNP)genotyping assay.Haplotype analysis was conducted using Haploview 4.2 software.Statistical analyses were performed using SPSS software,including analysis of variance(ANOVA),t-test,chi-square test,and logis-tic regression analysis.Results:The offspring of the longevous elderly had significantly higher MMSE scores(25.56±3.07)than the local elderly controls and non-local elderly controls,and exhibited a lower cognitive impair-ment rate(11.0%)compared with the other two groups(P＜0.05).In the dominant genetic model of the lon-gevous elderly,offspring of the longevous elderly,and local elderly controls,compared with the individuals carry-ing the TNF rs1799964 TT genotype,the individuals carrying the rs1799964 TC+CC genotype had a significantly reduced risk of cognitive impairment(P＜0.05).Compared with the individuals carrying the rs1800630_CC geno-type,the individuals carrying the rs1800630 CA+AA genotype had a significantly lower risk of cognitive impair-ment(P＜0.05).In the allelic model,the individuals carrying the rs1800630_A allele showed a lower incidence of cognitive impairment than those carrying the C allele(OR=0.341,0.311 and 0.457,respectively;P＜0.05).However,no association was found between the genotypes of rs1799964 and rs1800630 and cognitive function in the non-local elderly controls.Haplotype analysis revealed that the TNF C-A-C haplotype(rs1799964-rs1800630-rs1799724)was negatively correlated with the risk of cognitive impairment in the longevous elderly,offspring of the longevous elderly,and local elderly controls(OR=0.369,0.313 and 0.452,respectively;all P＜0.05),whereas no such correlation was observed in the non-local elderly controls.Conclusion:In the population of the Hongshui River Basin in Guangxi,the direct offsprings of longevous individuals maintain relatively good cogni-tive function.The individuals carrying the TNF rs1799964_C and rs1800630_A alleles have a significantly re-duced risk of cognitive impairment,serving as protective genetic factors against cognitive aging.
Objective To investigate the hematological parameters and clinical phenotypes of a heterozygote carrying the rare IVSⅠ-2 (T＞A) mutation in the β-globin gene. Methods Patients suspected of having β-thalassemia who underwent initial screening via routine blood tests [red blood cell count (RBC), hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), hematocrit (HCT), red blood cell distribution width (RDW)], and analyses of hemoglobin A2 (Hb A2) and fetal hemoglobin (Hb F) were enrolled in this study. Common β-thalassemia gene mutations were detected using fluorescent polymerase chain reaction (PCR) melting curve analysis. For individuals in whom no mutations were identified by the above methods, DNA sequencing was further performed to detect rare or unknown β-thalassemia gene mutations. Results Among 235 patients with β-thalassemia, 1 patient with rare β-thalassemia was identified. The patient harbored a heterozygous IVSⅠ-2 (T＞A) (HBB:c.92+2 T＞A) mutation in the β-globin gene coexisting with α-thalassemia of the αα/-α3.7 deletion type. The patient's routine blood test results were as follows: RBC 5.89×1012/L, Hb 107.00 g/L, MCV 55.40 fL, MCH 18.15 pg, MCHC 327.70 g/L, HCT 0.327 and RDW 0.18. The hemoglobin analysis results of the patient revealed an Hb A2 level of 5.0% and an Hb F level of 4.0%. Conclusion β-Thalassemia caused by the rare heterozygous IVS Ⅰ-2 (T＞A) mutation in the β-globin gene is reported for the first time in China. Clinically, the patient presents with mild anemia, decreased MCV and MCH levels, and elevated Hb A2. This mutation is relatively rare and prone to missed diagnosis.
Objective:To explore the associations between organochlorine pesticides(OCPs)and cardiovascular disease(CVD)in the general population.Methods:A cross-sectional study was adopted,and 6,009 residents who met the inclusion and exclusion criteria from the National Health and Nutrition Examination Survey(NHANES)(2005-2016)were enrolled.General data,physical examination,and laboratory test results were collected from the participants.Seven types of OCPs were analyzed,including β-hexachlorocyclohexane(β-HCH),hexachloro-benzene(HCB),oxychlordane(OXY),trans-nonachlor(T-NONA),2,2-Bis(4-chlorophenyl)-1,1-dichloroethene(p,p'-DDE),2,2-Bis(4-chlorophenyl)-1,1,1-trichloroethane(p,p'-DDT)and Mirex.The participants were divided into a control group(n=5,499)and a CVD group(n=510)according to the pre-sence or absence of CVD.Multi-variate logistic regression analysis and restricted cubic spline(RCS)models were used to analyze the associations between individual exposure to OCPs and CVD risk in the population.The associations between the joint expo-sure to OCPs and CVD risk were further analyzed using Bayesian kernel machine regression(BKMR)and quan-tile g-computation(Qgcomp)models.Results:Results from the multivariate logistic regression analysis showed that,after adjusting for confounders,elevated concentrations of β-HCH,OXY,T-NONA,p,p'-DDE and Mirex were associated with an increased risk of CVD(P＜0.05).Compared with the first quartile of concentration,the CVD risk associated with β-HCH,OXY,T-NONA,p,p'-DDE and Mirex at the fourth quartile concentration in-creased by 3.19-fold,2.96-fold,6.26-fold,2.40-fold and 1.89-fold,respectively.The RCS model revealed non-linear dose-response relationships between β-HCH,OXY,T-NONA,Mirex and CVD risk(P＜0.05).The results from the BKMR model indicated a positive joint effect of OCPs mixture on CVD risk,with OXY likely being one of the major contributors.When the concentrations of other OCPs were fixed at the 25th,50th,and 75th per-centiles,OXY was positively associated with CVD risk.Additionally,potential interactions were observed be-tween OXY and β-HCH,OXY and p,p'-DDE,as well as OXY and Mirex.The Qgcomp model verified the stabil-ity and validity of the BKMR,with only p,p'-DDE exerting a negative effect.The positive weights were observed in the following order:OXY,T-NONA,Mirex,and β-HCH.Conclusion:Joint exposure to OCPs is associated with an increased risk of CVD.OXY is the major component driving the associations,suggesting that it may serve as an independent risk factor for increased CVD risk.
Objective To analyze the expression and prognostic value of CTNNA1 in hepatocellular carcinoma (HCC), and to explore the impact of CTNNA1 on the malignant biological behavior of HCC cells. Methods Public databases and clinical specimens were utilized to analyze the expression level of CTNNA1 in HCC and its correlation with patient prognosis. Western blotting was used to detect the expression of CTNNA1 protein in HCC cell lines including Huh7, SK-HEP1, LM3, SNU449, and MHCC-97H. Lentiviral vectors were used to overexpress the CTNNA1 gene in SNU449 cells, and short hairpin RNA (shRNA) was applied to knock down CTNNA1 in Huh7 and MHCC-97H cells. Cell counting kit-8 (CCK-8) assay, plate colony formation assay, cell scratch assay, and Transwell migration and invasion assays were used to detect the proliferation, migration and invasion abilities of cells with CTNNA1 overexpression and knockdown. Results The analysis of UALCAN and GEPIA databases, combined with clinical sample validation, indicated that compared with adjacent normal tissues, the mRNA and protein expression of CTNNA1 were upregulated in HCC cancer tissues, which was associated with poor patient prognosis (P < 0.05). Compared with the control group, the proliferation, migration and invasion abilities of HCC cells were significantly enhanced after CTNNA1 overexpression in SNU449 cells. Conversely, knockdown of CTNNA1 expression in Huh7 and MHCC-97H cells markedly weakened these malignant biological abilities (P < 0.05). Conclusion CTNNA1 is upregulated in HCC, and its high expression is closely correlated with poor prognosis. CTNNA1 may serve as a potential prognostic biomarker and therapeutic target for HCC.
Objective To analyze the mutation types of non-deletional hemoglobin H (Hb H) disease, explore the correlation between distinct genotypes and clinical phenotypes, and identify rare α-globin gene mutations, so as to provide evidence for clinical diagnosis and treatment, genetic counseling, and prenatal diagnosis. Methods Routine blood tests [hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC)] and Hb H analysis were performed on patients diagnosed and treated for α-thalassemia at the First Affiliated Hospital of Guangxi Medical University from January 2024 to January 2026. Gap-polymerase chain reaction (gap-PCR), fluorescence-based PCR melting curve assay (FCMA) and DNA sequencing were used for genetic analysis of thalassemia. Results Among 217 patients with non-deletional Hb H disease, 187 patients were identified as Hb H-CS (--SEA/αCSα) and 27 patients as Hb H-QS(--SEA/αQSα), and 3 patients carried rare gene mutations causing Hb H disease, including one case each of --SEA/αATG＞GTGα, --SEA/αCD90-92(-AGCTTCGG)α and --SEA/αCD30(-GAG)α. None of the patients were complicated with β-thalassemia. The results of routine blood tests showed that mild, moderate and severe anemia in the Hb H-CS group accounted for 22.99%, 64.71% and 12.30%, respectively; the corresponding proportions of mild, moderate and severe anemia in the Hb H-QS group accounted for 44.45%, 51.85% and 3.70%, respectively. The Hb levels were 107.30 g/L for the --SEA/αATG＞GTGα genotype, 88.40 g/L for the --SEA/αCD90-92(-AGCTTCGG)α genotype, and 73.70 g/L for the --SEA/αCD30(-GAG)α genotype, all accompanied by decreased MCV and MCH. Hb analysis revealed that the Hb H level was 13.60% (10.45%–15.90%) in the Hb H-CS group and 23.20% (17.30%–25.00%) in the Hb H-QS group, with a statistically significant difference between the two groups (P＜0.05). The Hb H levels of the --SEA/αATG＞GTGα, --SEA/αCD90-92(-AGCTTCGG)α and --SEA/αCD30(-GAG)α genotypes were 25.30%, 24.40% and 20.40%, respectively. Conclusion The predominant genotype of non-deletional Hb H disease is --SEA/αCSα, followed by --SEA/αQSα. Moderate anemia is the main clinical manifestation of non-deletional Hb H disease. The Hb H level in the Hb H-QS group is higher than that in the Hb H-CS group. Three cases of Hb H disease with genotypes of --SEA/αATG＞GTGα, --SEA/αCD90-92(-AGCTTCGG)α and --SEA/αCD30(-GAG)α are identified, presenting with mild to moderate anemia.
Carbon-based dual-atom nanozymes(DAzymes)have emerged as a research hotspot in the field of nanozymes due to their highly controllable atomic configurations and superior electronic structure regulation capabilities.This paper systematically reviews the primary preparation methods for carbon-based DAzymes and compares their advantages.Centering on biomedical application scenarios,it also summarizes the functional advantages and application performances of carbon-based DAzymes.Finally,it outlines the future development trends in data-driven design,multifunctional integration,and clinical translation,aiming to provide insights for the rational design and application expansion of carbon-based DAzymes.
Thalassemia (Thal) is a major hereditary birth defect disease in southern China. As a high-prevalence region for Thal, Guangxi has significantly reduced the birth rate of infants with thalassemia major through a government-led and multi-departmental prevention system integrating premarital and preconception screening, prenatal diagnosis, genetic counseling, and standardized intervention. This experience provides an important model for comprehensive Thal prevention and control. At present, the prevention and control of thalassemia major has entered a new stage focusing on long-term consolidation and quality improvement rather than rapid reduction of affected births alone. Future efforts should continue to move the prevention gateway forward, refine an integrated screening system covering the premarital, preconception, and early pregnancy periods, strengthen closed-loop management of screen-positive couples, improve laboratory quality control, enhance genetic counseling capacity, and address regional disparities. Meanwhile, attention should be paid to comprehensive treatment for children already born with thalassemia major, including standardized blood transfusions, iron chelation therapy, hematopoietic stem cell transplantation, and gene therapy, thereby shifting the prevention-and-control system from "preventing new cases" to a model of "preventing new cases, managing existing cases, and pursuing cure". The experience from Guangxi suggests that only the integration of public-health-oriented prevention and clinical treatment can sustainably consolidate the achievements in prevention and control of thalassemia major, and provide a replicable pathway for the comprehensive prevention and treatment of birth defects in China."preventing new cases" to a model of "preventing new cases, managing existing cases, and pursuing cure". The experience from Guangxi suggests that only the integration of public-health-oriented prevention and clinical treatment can sustainably consolidate the achievements in prevention and control of thalassemia major, and provide a replicable pathway for the comprehensive prevention and treatment of birth defects in China.
Objective:To investigate the effects of Avicennia marina leaf-derived extracellular vesicles(AmLEVs)on skin wound healing in diabetic rats and explore the underlying mechanisms.Methods:Extracellular vesicles were isolated from Avicennia marina leaves by differential centrifugation combined with ultracentrifugation.The morphology of the extracellular vesicles was observed by transmission electron microscopy(TEM),and the par-ticle size and zeta potential of AmLEVs were measured by dynamic light scattering(DLS)using a Malvern instru-ment.The antioxidant capacity of AmLEVs was evaluated using DPPH,ABTS,and hydroxyl radical assay kits.Human umbilical vein endothelial cells(HUVECs)and human skin fibroblasts(HSFs)were selected as experi-mental cells,and a high-glucose medium containing 25 mmol/L glucose was used to mimic the diabetic microen-vironment.After co-culture with AmLEVs,the levels of intracellular reactive oxygen species(ROS)were detected using the DCFH-DA probe.Cell viability,cell migration,and angiogenic capacity were assessed by cell counting kit-8(CCK-8)assay,scratch wound assay,and tube formation assay,respectively.The expression levels of VEGF mRNA and VEGF protein were determined by real-time quantitative PCR(RT-qPCR)and immunofluo-rescence staining.In addition,a full-thickness skin wound model was established in streptozotocin-induced dia-betic Sprague-Dawley(SD)rats to evaluate the wound-healing effects of AmLEVs.Results:AmLEVs were suc-cessfully isolated,with a typical cup-shaped morphology of the nanovesicles and an average particle size of ap-proximately 55 nm.In vitro experiments showed that AmLEVs exhibited strong scavenging activities against DPPH,ABTS,and hydroxyl radicals.Moreover,100 μg/mL AmLEVs effectively reduced high glucose-induced ROS accumulation,thereby reversing the inhibitory effects of the hyperglycemic environment on the prolifera-tion,migration,and tube formation of HUVECs,thus enhancing the viability of HSFs.Compared with the control group,the mRNA and protein expression levels of VEGF were decreased in the high-glucose injury group.How-ever,these levels were increased when the high-glucose injury group was treated with 100 μg/mL AmLEVs.In vivo experiments demonstrated that the wound healing rate in the rats of diabetic group was significantly decreased compared with that in the rats of the control group.However,the wound healing rate was significantly increased when the diabetic rats were treated with AmLEVs.HE and Masson staining revealed that AmLEV treat-ment enhanced re-epithelialization and increased collagen deposition compared with the diabetic model group.Furthermore,immunofluorescence staining,RT-qPCR,and western blotting confirmed that VEGF mRNA and protein expressions in skin tissues were downregulated in diabetic rats but were significantly upregulated follow-ing AmLEV treatment.Conclusion:AmLEVs effectively promote diabetic wound healing.The underlying mechanisms may involve modulating cellular oxidative stress,enhancing angiogenesis,and improving cell migra-tion,suggesting that AmLEVs can serve as a potential therapeutic agent for diabetic wound treatment.
Ovarian aging drives not only the progressive decline of female reproductive function but also the acceleration of multi-systemic aging. Aberrant extracellular matrix deposition and the consequent biomechanical changes in tissues disrupt the peri-follicular microenvironment, thereby contributing to both natural ovarian aging and premature ovarian insufficiency. This paper systematically reviews the mechanisms underlying ovarian fibrosis, integrating recent insights from single-cell and spatial multi-omics analyses to delineate the characteristics of the fibrotic and inflammatory microenvironment, summarizes the clinical potential of non-invasive assessment modalities, including shear wave elastography and humoral molecular markers, and evaluates current anti-fibrotic therapeutic strategies, aiming to provide valuable insights and directions for the early identification and clinical intervention of ovarian aging-related disorders.
Objective:To explore the neuroprotective effect and mechanism of soy isoflavones(SI)on cerebral ischemia-reperfusion injury(CIRI)by inhibiting NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome-mediated neuronal pyroptosis.Methods:Sixty healthy Sprague-Dawley(SD)rats were randomly divided into sham group,model group,SI group and SI+Nigericin group.Rats in the SI group were pre-treated with intragastric administration of SI(120 mg/kg)once daily for 21 consecutive days.The middle cerebra-lartery occlusion(MCAO)rat model was established by the suture-occluded method,and reperfusion was in-duced by withdrawing the suture after 2 hours of ischemia;the sham group only underwent vascular separation without suture insertion.Five minutes before reperfusion,rats in the SI+Nigericin group were injected with Nige-ricin(1 mg/kg)via the tail vein.Twenty-four hours after reperfusion,the severity of brain injury was assessed by neurological deficit score,brain water content(dry-wet weight method),2,3,5-triphenyltetrazolium chloride(TTC)staining,and hematoxylin-eosin(HE)staining.The expression of pyroptosis-related proteins was mea-sured by immunofluorescence staining and western blotting.The serum levels of interleukin(IL)-1β,IL-18 and the activity of lactate dehydrogenase(LDH)were measured by enzyme-linked immunosorbent assay(ELISA).Results:Compared with the model group,the SI group exhibited significantly reduced neurological deficit scores,brain water content,cerebral infarct volume(P＜0.001),as well as obviously alleviated cortical patho-logical injury.In the SI group,the protein expression levels of NLRP3,Gasdermin D(GSDMD),GSDMD N-terminal domain(GSDMD-N),caspase-1,cleaved caspase-1,IL-18,IL-1β and IL-1β p17 were downregulated;se-rum IL-1β and IL-18 contents as well as LDH activity were reduced(all P＜0.05),and the fluorescence signals of NLRP3 and GSDMD-N in brain tissues were obviously weakened.Nigericin could partially reverse the above protective effects of SI(P＜0.05).Conclusion:SI reduces neuronal pyroptosis by inhibiting NLRP3 inflamma-some activation,thereby effectively alleviating CIRI and exerting neuroprotective effects.
