Objective To investigate the relationship between serum levels of plasminogen activator inhibi-tor-1(PAI-1)and annexin A2(ANXA2)and the carotid atherosclerosis stability(CAS)plaque in patients with transient ischemic attack(TIA).Methods A total of 131 patients with TIA admitted to the hospital from January 2021 to January 2023 were selected as the TIA group,and 46 healthy people in the same period were selected as the control group.According to the CAS plaque stability of TIA patients,they were divided into unstable plaque group(64 cases)and stable plaque/no plaque group(67 cases).The serum levels of PAI-1 and ANXA2 were detected by enzyme-linked immunosorbent assay and chemiluminescence immunoassay,respectively.Multivariate Logistic regression analysis was used to analyze the factors of CAS plaque stability in TIA patients.Receiver operating characteristic curve was used to analyze the predictive value of serum PAI-1 and ANXA2 levels for CAS instability in TIA patients.Results Compared with the control group,the levels of PAI-1 and ANXA2 in TIA group were significantly increased(P<0.05).Compared with the stable plaque/no plaque group,the unstable plaque group was significant increaseed in the serum levels of PAI-1 and ANXA2(P<0.05).Smoking,high risk of TIA,elevated PAI-1 and ANXA2 were independent risk factors for CAS plaque stability in patients with TIA(P<0.05).The area under the curve predicted by serum PAI-1 and ANXA2 levels combined was 0.879,which was larger than 0.788 and 0.783 predicted by serum PAI-1 and ANXA2 levels alone(P<0.05).Conclusion The increased levels of serum PAI-1 and ANXA2 are closely re-lated to CAS plaque instability in patients with TIA.The combination of serum PAI-1 and ANXA2 levels has a higher value in predicting CAS plaque instability in patients with TIA.

Objective:To investigate the biological role of miR-1224-5p in the proliferation,migration and apoptosis of prostate cancer cells,as well as its regulatory mechanism of expression.Methods:Human prostate cancer cells(PC3,DU145,LNCaP,22Rv1)and normal prostate epithelial RWPE-1 cells were selected for this study.The expression of miR-1224-5p in prostate cancer cells was detected by qPCR.miR-1224-5p mimic,inhibitor and corresponding negative control(NC)plasmids were transfected into PC3 and DU145 cells by liposome transfection technology,respectively.The transfection efficiency was verified by qPCR.The effects of miR-1224-5p mimic and inhibitor on cell proliferation,migration and apoptosis were detected by CCK-8 assay,plate clone formation assay,scratch healing assay and flow cytometry.The potential N6-methyladenosine(m6A)modification sites in the pri-miR-1224-5p sequence were predicted using SRAMP website,and the prediction results were verified using methylated RNA immunoprecipitation(MeRIP).The expression of methyltransferase 3(METTL3),a key methyltransferase involved in m6A modification,in prostate cancer cells was detected by qPCR.CCK-8 assay and Transwell assay were used to detect the effect of METTL3 siRNA transfection on the proliferation and migration of PC3 and DU145 cells.The regulatory effect of METTL3-mediated m6A modification on the expression of miR-1224-5p was detected by qPCR and MeRIP.Results:miR-1224-5p was upregulated in prostate cancer cells(all P<0.01).Transfection with miR-1224-5p mimic promoted the proliferation,migration and inhibited the apoptosis of PC3 and DU145 cells(P<0.05 or P<0.01).Conversely,miR-1224-5p inhibitor suppressed the proliferation,migration and induced apoptosis of PC3 and DU145 cells(P<0.05 or P<0.01).pri-miR-1224-5p contained m6A modification sites.METTL3 was highly expressed in prostate cancer cells(all P<0.01).Transfection of METTL3 siRNA inhibited the proliferation and migration of PC3 and DU145 cells(all P<0.01).METTL3-mediated m6A modification regulated the expression of miR-1224-5p.Conclusion:miR-1224-5p is upregulated in prostate cancer cells through METTL3-mediated m6A modification.Down-regulation of miR-1224-5p can inhibit the proliferation,migration and induce apoptosis of prostate cancer cells.

In the context of global climate change, the impact of ambient temperature change on cardiovascular health has become increasingly significant. Epidemiological studies have shown that both high and low temperature can lead to the occurrence of adverse cardiovascular outcomes. For example, exposure to low temperature and high temperature, and diurnal temperature difference can increase the risk of cardiovascular diseases (CVD) mortality. Exposure to cold and heat, heatwaves and cold spells may induce myocardial infarction. Low temperature, high temperature and variations in temperature can increase the risk of heart failure. There are complex interactions among the effects of temperature and humidity, wind speed, radiation, precipitation and air pollution on adverse cardiovascular outcomes. Valnerable groups such as the elderly, patients with chronic diseases, and outdoor workers are particularly susceptible to temperature-related CVD risks. The mechanisms by which temperature affects adverse cardiovascular outcomes involves hemodynamic changes, autonomic nervous system dysregulation, blood rheology alterations, as well as inflammation and metabolic abnormalities, but many research gaps remain. Future studies should conduct in-depth research on mechanisms, individual differences, climate change impacts, and public health interventions to provide more effective policies and interventions to reduce the burden of CVD.

Objective To evaluate the clinical efficacy of stellate ganglion block(SGB)combined with intranasal dexmedetomidine for the treatment of insomnia.Methods A total of 64 patients aged 18 to 75 with insom-nia were randomly assigned to either the experimental group(DS group)or the control group(S group).The S group received SGB treatment for 14 consecutive days,whereas the DS group received an additional intranasal dexmedeto-midine spray at a dose of 100 μg,administered 30 minutes before bedtime on days 1 through 6,in conjunction with SGB.We measured and recorded the Pittsburgh Sleep Quality Index(PSQI),Self-Rating Depression Scale(SDS),Self-Rating Anxiety Scale(SAS)scores,and Psychomotor Vigilance Test(PVT)results for both groups at three time points:baseline(T1),the day after treatment(T2),and one month after treatment(T3).Results Intra-group Com-parison:In both the DS and S groups,PSQI scores and dimensionspecific scores at T2 and T3 were significantly lower compared to T1(P<0.05).SAS and SDS scores in both groups showed a significant reduction at T3 compared to T1 and T2(P<0.05),while PVT results exhibited no significant changes(P>0.05).Inter-group Comparison:The PSQI scores and dimensionspecific scores in the DS group at T2(8.44±2.99)and T3(8.22±2.60)were significantly lower than those in the S group at T2(10.88±2.56)and T3(10.88±2.84)(P<0.05).However,no significant differences were observed in SDS and SAS scores between the DS and S groups at T2 and T3(P>0.05).Conclusion Compared to standalone SGB,the combination of SGB with intranasal dexmedetomidine significantly enhances sleep quality in patients with insomnia,while not impacting their levels of anxiety,depression,or alertness.

Objective:To investigate the association between dopamine D2 receptor ( DRD2) promoter methylation levels and melancholic depression (MD), and its correlation with psychosocial factors and depression severity. Method:A total of 30 patients with melancholic depression (MD group) and 30 with non-melancholic depression (NMD group) were recruited from the First Affiliated Hospital of Xinjiang Medical University between October 2023 and October 2024, along with 30 healthy controls. Participants were assessed by the Hamilton depression rating scale (HAMD-24), dimensional anhedonia rating scale (DARS), childhood trauma questionnaire (CTQ), life event scale (LES), and social support rating scale (SSRS). DRD2 methylation levels in peripheral blood were measured by MassArray technology. Data were analyzed in SPSS 29.0 software using ANOVA, non-parametric tests, and t-tests to compare methylation levels among groups, and Logistic regression, Pearson correlation, and multiple linear regression to examine associations between DRD2 methylation and psychosocial factors and depression severity. Results:(1) There were statistically significant differences among the three groups in years of education ( F=9.873, P=0.007), HAMD-24 total scores ( H=76.669, P<0.001), DARS total scores ( H=60.617, P<0.001), SSRS total scores ( F=3.592, P=0.032), LES negative stimulus scores ( H=43.461, P<0.001), and CTQ total scores ( H=10.751, P=0.005). The MD group had a longer course of illness ( Z=-1.980, P=0.048), more episodes ( Z=-2.027, P=0.043), and a higher rate of suicide attempts in the past year ( χ2= 9.643, P=0.002) compared with the NMD group. The MD group had higher HAMD-24 total scores (31.00 (28.00, 32.25)) than the NMD group (23.00 (21.00, 25.00)), and lower DARS total scores (17.00 (9.75, 22.00)) compared with the NMD group(36.50 (33.75, 43.00)) (both P<0.05). (2) No statistically significant difference was found in DRD2 gene methylation levels among the three groups ( F=0.081, all P>0.05). Logistic regression showed that DRD2 gene methylation level was not an influencing factor for the onset of MD ( P>0.05). (3)The correlation analysis and multiple linear regression analysis showed that childhood trauma (CTQ total score) was negatively correlated with DRD2 gene methylation levels in MD patients ( r=-0.416, P=0.025) and MDD patients. DRD2 gene methylation level was not correlated with depression severity ( r=0.136, P>0.05) or anhedonia severity (both P>0.05). Social support (SSRS total score) was positively correlated with DARS total scores in MD patients( r=0.427, P=0.019), and years of education had a positive association with DARS total scores in MDD patients ( B=1.527, P=0.030), suggesting that both are negatively correlated with the degree of anhedonia. Conclusion:DRD2 methylation may be influenced by early-life trauma, but its role in the pathogenesis of MD and its association with depression severity require further investigation.

Objective:To investigate the association between dopamine D2 receptor ( DRD2) promoter methylation levels and melancholic depression (MD), and its correlation with psychosocial factors and depression severity. Method:A total of 30 patients with melancholic depression (MD group) and 30 with non-melancholic depression (NMD group) were recruited from the First Affiliated Hospital of Xinjiang Medical University between October 2023 and October 2024, along with 30 healthy controls. Participants were assessed by the Hamilton depression rating scale (HAMD-24), dimensional anhedonia rating scale (DARS), childhood trauma questionnaire (CTQ), life event scale (LES), and social support rating scale (SSRS). DRD2 methylation levels in peripheral blood were measured by MassArray technology. Data were analyzed in SPSS 29.0 software using ANOVA, non-parametric tests, and t-tests to compare methylation levels among groups, and Logistic regression, Pearson correlation, and multiple linear regression to examine associations between DRD2 methylation and psychosocial factors and depression severity. Results:(1) There were statistically significant differences among the three groups in years of education ( F=9.873, P=0.007), HAMD-24 total scores ( H=76.669, P<0.001), DARS total scores ( H=60.617, P<0.001), SSRS total scores ( F=3.592, P=0.032), LES negative stimulus scores ( H=43.461, P<0.001), and CTQ total scores ( H=10.751, P=0.005). The MD group had a longer course of illness ( Z=-1.980, P=0.048), more episodes ( Z=-2.027, P=0.043), and a higher rate of suicide attempts in the past year ( χ2= 9.643, P=0.002) compared with the NMD group. The MD group had higher HAMD-24 total scores (31.00 (28.00, 32.25)) than the NMD group (23.00 (21.00, 25.00)), and lower DARS total scores (17.00 (9.75, 22.00)) compared with the NMD group(36.50 (33.75, 43.00)) (both P<0.05). (2) No statistically significant difference was found in DRD2 gene methylation levels among the three groups ( F=0.081, all P>0.05). Logistic regression showed that DRD2 gene methylation level was not an influencing factor for the onset of MD ( P>0.05). (3)The correlation analysis and multiple linear regression analysis showed that childhood trauma (CTQ total score) was negatively correlated with DRD2 gene methylation levels in MD patients ( r=-0.416, P=0.025) and MDD patients. DRD2 gene methylation level was not correlated with depression severity ( r=0.136, P>0.05) or anhedonia severity (both P>0.05). Social support (SSRS total score) was positively correlated with DARS total scores in MD patients( r=0.427, P=0.019), and years of education had a positive association with DARS total scores in MDD patients ( B=1.527, P=0.030), suggesting that both are negatively correlated with the degree of anhedonia. Conclusion:DRD2 methylation may be influenced by early-life trauma, but its role in the pathogenesis of MD and its association with depression severity require further investigation.

Objective To evaluate the clinical efficacy of stellate ganglion block(SGB)combined with intranasal dexmedetomidine for the treatment of insomnia.Methods A total of 64 patients aged 18 to 75 with insom-nia were randomly assigned to either the experimental group(DS group)or the control group(S group).The S group received SGB treatment for 14 consecutive days,whereas the DS group received an additional intranasal dexmedeto-midine spray at a dose of 100 μg,administered 30 minutes before bedtime on days 1 through 6,in conjunction with SGB.We measured and recorded the Pittsburgh Sleep Quality Index(PSQI),Self-Rating Depression Scale(SDS),Self-Rating Anxiety Scale(SAS)scores,and Psychomotor Vigilance Test(PVT)results for both groups at three time points:baseline(T1),the day after treatment(T2),and one month after treatment(T3).Results Intra-group Com-parison:In both the DS and S groups,PSQI scores and dimensionspecific scores at T2 and T3 were significantly lower compared to T1(P<0.05).SAS and SDS scores in both groups showed a significant reduction at T3 compared to T1 and T2(P<0.05),while PVT results exhibited no significant changes(P>0.05).Inter-group Comparison:The PSQI scores and dimensionspecific scores in the DS group at T2(8.44±2.99)and T3(8.22±2.60)were significantly lower than those in the S group at T2(10.88±2.56)and T3(10.88±2.84)(P<0.05).However,no significant differences were observed in SDS and SAS scores between the DS and S groups at T2 and T3(P>0.05).Conclusion Compared to standalone SGB,the combination of SGB with intranasal dexmedetomidine significantly enhances sleep quality in patients with insomnia,while not impacting their levels of anxiety,depression,or alertness.

Animal experiments are widely used in biomedical research for safety assessment,toxicological analysis,efficacy evaluation,and mechanism exploration.In recent years,the ethical review system has become more stringent,and awareness of animal welfare has continuously increased.To promote more efficient and cost-effective drug research and development,the United States passed the Food and Drug Administration(FDA)Modernization Act 2.0 in September 2022,which removed the federal mandate requiring animal testing in preclinical drug research.In April 2025,the FDA further proposed to adopt a series of"new alternative methods"in the research and development of drugs such as monoclonal antibodies,which included artificial intelligence computing models,organoid toxicity tests,and 3D micro-physiological systems,thereby gradually phasing out traditional animal experiment models.Among these cutting-edge technologies,3D bioprinting models are a significant alternative and complement to animal models,owing to their high biomimetic properties,reproducibility,and scalability.This review provides a comprehensive overview of advancements and applications of 3D bioprinting technology in the fields of biomedical and pharmaceutical research.It starts by detailing the essential elements of 3D bioprinting,including the selection and functional design of biomaterials,along with an explanation of the principles and characteristics of various printing strategies,highlighting the advantages in constructing complex multicellular spatial structures,regulating microenvironments,and guiding cell fate.It then discusses the typical applications of 3D bioprinting in drug research and development,including high-throughput screening of drug efficacy by constructing disease models such as tumors,infectious diseases,and rare diseases,as well as conducting drug toxicology research by building organ-specific models such as those of liver and heart.Additionally,the review examines the role of 3D bioprinting in tissue engineering,discussing its contributions to the construction of functional tissues such as bone,cartilage,skin,and blood vessels,as well as the latest progress in regeneration and replacement.Furthermore,this review analyzes the complementary advantages of 3D bioprinting models and animal models in the research of disease progression,drug mechanisms,precision medicine,drug development,and tissue regeneration,and discusses the potential and challenges of their integration in improving model accuracy and physiological relevance.In conclusion,as a cutting-edge in vitro modeling and manufacturing technology,3D bioprinting is gradually establishing a comprehensive application system covering disease modeling,drug screening,toxicity prediction,and tissue regeneration.

Objective     To explore the correlation between the quantitative and qualitative features of CT images and the invasiveness of pulmonary ground-glass nodules, providing reference value for preoperative planning of patients with ground-glass nodules. Methods    The patients with ground-glass nodules who underwent surgical treatment and were diagnosed with pulmonary adenocarcinoma from September 2020 to July 2022 at the Third Affiliated Hospital of Kunming Medical University were collected. Based on the pathological diagnosis results, they were divided into two groups: a non-invasive adenocarcinoma group with in situ and minimally invasive adenocarcinoma, and an invasive adenocarcinoma group. Imaging features were collected, and a univariate logistic regression analysis was conducted on the clinical and imaging data of the patients. Variables with statistical difference were selected for multivariate logistic regression analysis to establish a predictive model of invasive adenocarcinoma based on independent risk factors. Finally, the sensitivity and specificity were calculated based on the Youden index. Results     A total of 555 patients were collected. The were 310 patients in the non-invasive adenocarcinoma group, including 235 females and 75 males, with a meadian age of 49 (43, 58) years, and 245 patients in the invasive adenocarcinoma group, including 163 females and 82 males, with a meadian age of 53 (46, 61) years. The binary logistic regression analysis showed that the maximum diameter (OR=4.707, 95%CI 2.060 to 10.758), consolidation/tumor ratio (CTR, OR=1.027, 95%CI 1.011 to 1.043), maximum CT value (OR=1.025, 95%CI 1.004 to 1.047), mean CT value (OR=1.035, 95%CI 1.008 to 1.063), spiculation sign (OR=2.055, 95%CI 1.148 to 3.679), and vascular convergence sign (OR=2.508, 95%CI 1.345 to 4.676) were independent risk factors for the occurrence of invasive adenocarcinoma (P<0.05). Based on the independent predictive factors, a predictive model of invasive adenocarcinoma was constructed. The formula for the model prediction was: Logit(P)=–1.293+1.549×maximum diameter of lesion+0.026×CTR+0.025×maximum CT value+0.034×mean CT value+0.72×spiculation sign+0.919×vascular convergence sign. The area under the receiver operating characteristic curve of the model was 0.910 (95%CI 0.885 to 0.934), indicating that the model had good discrimination ability. The calibration curve showed that the predictive model had good calibration, and the decision analysis curve showed that the model had good clinical utility. Conclusion     The predictive model combining quantitative and qualitative features of CT has a good predictive ability for the invasiveness of ground-glass nodules. Its predictive performance is higher than any single indicator.

Objective To explore the mechanism of ferroptosis induced by endoplasmic reticulum stress(ERs)in acute respiratory distress syndrome(ARDS).Methods In order to determine the effects of LPS on oxidative stress and Fe2+level of mouse capillary alveolar epithelial cells(MLE12 cells),the cells were treated with LPS(0,1,2,5 μg/ml)for 24 h.To verify the role of ferroptosis in lipopolysaccharide(LPS)-induced cell death,MLE12 cells were divided into control(Con)group,iron removal inhibitor(Fer-1)group,LPS group and LPS+Fer-1 group.LPS+Fer-1 group was pretreated with 10 μmol/L Fer-1 for 6 h,then the cells were exposed to 5 μg/ml LPS for 24 h.Con group was treated with solvent DMSO for 24 h.Fer-1 group was pretreated with 10 μmol/L Fer-1 for 6 h,and then treated with DMSO for 24 h.The cells in LPS group were exposed to 5 μg/ml LPS for 24 h.The MLE12 cells were divided into three groups:Con+Vector group,Con+sequence similarity family 134 mem-ber B(FAM134B)group,LPS+Vector group and LPS+FAM134B group.After transfected with vector or FAM134B overexpression plasmid for 48 h,the cells were exposed or not exposed to 5 μg/ml LPS for 24 h.Cell vi-ability was measured by CCK-8.The levels of malondialdehyde(MDA),glutathione and iron,the protein levels of ferroptosis markers[cyclooxygenase 2(PTGS2),glutathione peroxidase 4(GPX4)]and ERs markers[glucose reg-ulatory protein 78(GRP78),activated transcription factor 4(ATF4)and C/EBP homologous protein(CHOP)]were measured in different groups.In order to further confirm the results of in vitro cell experiments,40 mice were randomly divided into Con+Vector group,Con+FAM134B group,LPS+Vector group and LPS+FAM134B group,with 10 mice in each group.LPS-induced sepsis models were established in LPS+Vector group and LPS+FAM134B group,and the levels of GPX4 and ERs in lung tissue were evaluated by immunofluorescence staining and protein blot.Results LPS treatment increased the levels of PTGS2 and MDA,and decreased the levels of GPX4 and GSH in MLE12 cells in a dose-dependent manner.Compared with LPS group,the cell viability,GPX4 and GSH levels in LPS+Fer-1 group increased significantly(P<0.05),while the PTGS2 protein level and MDA level decreased significantly(P<0.05).Compared with LPS+Vector group,LPS+FAM134B group significantly increased cell viability(P<0.05),decreased PTGS2 protein level(P<0.05)and increased GPX4 level(P<0.05).At the same time,the level of MDA in LPS+FAM134B group was lower than that in LPS+Vector group(P<0.05),and the level of GSH was higher than that in LPS+Vector group(P<0.05).In animal experiment,compared with LPS+Vector group,the expression levels of 4-HNE,ATF4 and CHOP in lung tissue of LPS+FAM134B group decreased significantly(P<0.05),and the expression levels of GPX4,FAM134B group in-creased significantly(P<0.05).Conclusion LPS induces ferroptosis and ERs in MLE12 cells in a dose-depend-ent manner.Activating the endoplasmic reticulum autophagy associated FAM134B receptor helps to inhibit ERs and alleviate cell ferroptosis.