Atherosclerotic cardiovascular disease(ASCVD)is one of the leading causes of death worldwide,and its progression is closely related to the pathological effects of neutrophil extracellular traps(NETs).In atherosclerosis(AS),NETs aggravate the process of disease by promoting inflammatory response,inducing endothelial dysfunction,promoting thrombosis and other mechanisms.The components such as myeloperoxidase(MPO),neutrophil elastase(NE)and citrullinated histone H3(CitH3)released by NETs can activate the immune inflammatory cascade,directly damage the vascular endothelium and promote thrombosis.In vascular inflammation,the formation of NETs is regulated by actin,and the released harmful molecules can induce endothelial cell apoptosis and drive the progress of inflammation through oxidative stress.The degradation and clearance of NETs depend on the action of enzymes such as deoxyribonuclease Ⅰ(DNase Ⅰ),and its regulatory mechanisms in atherosclerosis and vascular inflammation remain to be further studied.Based on the above mechanism,NETs-related markers have shown the potential as novel diagnostic and prognostic assessment biomarkers for ASCVD.This article aims to systematically elaborate the core pathological mechanism of NETs driving ASCVD through inflammatory activation,endothelial injury and thrombosis,providing a theoretical basis for targeted intervention.

Objective To investigate the susceptibility and infection characteristics of dengue virus(DENV)in cells derived from diverse tissues of tree shrews and to provide a basis for expanding the repertoire of DENV-permissive cell models in this species.Methods DENV was inoculated at a multiplicity of infection(MOI)of 0.02 into tree shrew skin fibroblasts(TSFs),primary tree shrew renal epithelial cells(pTRECs),tree shrew aortic endothelial cells(TAECs),tree shrew aortic smooth muscle cells(TASMCs),tree shrew hepatocytes(THs),tree shrew corneal stromal cells(TCSCs),tree shrew brain microvascular endothelial cells(TBMECs),and tree shrew retinal microvascular endothelial cells(TRMECs).C6/36,Vero,A549,and BHK-21 cells(commonly used for DENV propagation)were used as positive controls.Over 6 days post-infection,cellular cytopathic effects were monitored at 12-hour intervals using an inverted microscope,viral RNA loads in cell lysates were quantified by real-time fluorescent quantitative PCR to generate proliferation curves,and viral titers were determined by plaque assay.Results Seven types of tree shrew cells,except TRMECs,were susceptible to DENV.Prolonged infection induced pronounced cytopathic effects,including cell rounding,detachment,necrosis,and lysis,across all susceptible cells.The viral RNA loads detected in lysates of pTRECs,TBMECs,TASMCs,TAECs and THs,approached those of positive controls(≥4×107 copies/μL).Infectious progeny viruses were produced by these five cell types,with three(TAECs,3.13×105 PFU/mL;THs,2.03×105 PFU/mL;pTRECs,1.58×105 PFU/mL)exhibiting titers comparable to C6/36(3.85×10 5 PFU/mL)and earlier viral harvests.Conclusion DENV exhibits broad susceptibility to tree shrew cells of multiple tissue origins,with proliferation rates surpassing those of conventional cell lines sourced from other species.TAECs,THs,and pTRECs are particularly suitable for large-scale DENV proliferation,suggesting their potential involvement in in vivo infection.

With technological advancement and the growing demand for renewable energy, research and applications of new energy materials are becoming increasingly widespread. Typical new energy materials include lithium-ion battery materials, nanomaterials, nuclear energy materials and magnetic materials, etc., each of which has special toxicological characteristics. These materials may pose potential toxicological risks in the process of resource exploitation, production, transportation, usage, recycling or disposal, which have negatively impact on human health and the ecological environment. Occupational exposure is the main route of energy materials exposure, with potential health hazards on workers during the processes of production, transportation, recycling, and disposal. Among them, the disassembled batteries in the recycling or waste disposal process requires quality control, which is the high-risk position of occupational hazards. At present, the toxicology study of typical new energy materials mainly focuses on the potential impact of lithium-ion battery materials and nanomaterials on human health and the environment, but there are still limitations and challenges. In the future, it is necessary to further strengthen the human health risk management and prevention and control of new energy materials to protect human health and sustainable development.

Objective To investigate the association of spleen volume with the risk of non-alcoholic fatty liver disease(NAFLD)as well as their causal relationship.Methods We included 90 NAFLD cases and 47 healthy controls who had received contrast-enhanced computed tomography(CT)scan of the abdomen at the Second Affiliated Hospital of Xi'an Jiaotong University from November 2022 to November 2023.We conducted three-dimensional reconstruction of the spleen through a deep learning network model using a two-stage coarse-to-fine segmentation approach.We compared the two groups using the two-sample t test or Mann-Whitney U test for continuous data and using the chi-square test for categorical data;evaluated the correlation between spleen volume and liver function indicators through Pearson correlation or Spearman rank correlation analyses;determined the factors influencing the development of NAFLD through multivariable Logistic regression analysis;and further assessed the casual relationship between spleen volume and NAFLD using the inverse variance-weighted two-sample Mendelian randomization(IVW-MR)method.Results Spleen volume was significantly larger in NAFLD cases than in controls(272.93±104.16 vs 204.37±81.20 cm3,P<0.001).The Spearman rank correlation analysis showed that spleen volume was positively correlated with the hepatic steatosis index(rs=0.422,P<0.001)and gamma-glutamyl transferase levels(rs=0.211,P=0.047)in patients with NAFLD.The multivariable Logistic regression analysis indicated that spleen volume was an independent risk factor for the development of NAFLD(odds ratio[OR]=1.01,95%confidence interval[CI]:1.00-1.02,P=0.049).The IVW-MR analysis detected a causal relationship between spleen volume and NAFLD(OR=1.16,95%CI:1.05-1.28,P=0.005).Conclusion Increased spleen volume may be a risk factor for the development and progression of NAFLD.Further studies are still needed to investigate the specific mechanism.

Objective: To investigate the effects and mechanisms of Astragalus Polysacharin(APS) on the proliferation and metastasis of breast cancer cells by regulating miR-107 and miR-346-mediated macrophage polarization in breast cancer-derived exosomes. Methods: Forty 8-week-old female BALB/c mice were selected and breast cancer xenograft models and 4T1 transplanted tumor models were established. The mice were divided into the control group and the APS group. The APS group mice received daily intragastric administration of APS for 25 days, while the control group mice were given the same amount of normal saline. After all treatments were completed, the mice were euthanized, and tumor tissues were isolated. Western blot and flow cytometry were used to detect the expressions of proliferating cell nuclear antigen(PCNA), Ki-67, CD206, CD163, inducible nitric-oxide synthase(iNOS), and CD86. The apoptosis of single-cell suspensions in tumor tissues was analyzed. Human breast cancer cell line MDA-MB-231 was cultured and stimulated with APS, and exosomes from the cell culture medium were collected. The proliferation, migration, and invasion of cells were detected by CCK-8 assay, scratch assay, permeability chamber cell invasion assay, and qRT-PCR. Differentially expressed genes were screened by bioinformatics. Results : By measuring the expressions of molecules related to breast cancer cell proliferation and metastasis, it was shown that APS treatment reduced the expressions of proliferation-related proteins(PCNA and Ki-67) and metastasis-related proteins(Vimentin) in MDA-MB-231 xenograft tumor tissues; and the polarization of tumor-associated macrophages was observed. APS treatment of 4T1 transplanted tumor tissues could reduce the number of M2 macrophages and increase the number of M1 macrophages, resulting in a decrease in the ratio of M2/M1 macrophages and an increase in cell apoptosis in 4T1 transplanted tumor tissues. The expressions of related proteins iNOS and CD86 increased, and CD206 and CD163 decreased. After APS treatment, the exosomes produced by MDA-MB-231 reduced the polarization of M2 macrophages and affected the expressions of miR-107 and miR-346. Conclusion APS inhibits the polarization of M2 macrophages by regulating the expression of miR-107 or miR-346 in breast cancer cell-derived exosomes, ultimately inhibiting the proliferation and metastasis of breast cancer cells.

Atherosclerotic cardiovascular disease(ASCVD)is one of the leading causes of death worldwide,and its progression is closely related to the pathological effects of neutrophil extracellular traps(NETs).In atherosclerosis(AS),NETs aggravate the process of disease by promoting inflammatory response,inducing endothelial dysfunction,promoting thrombosis and other mechanisms.The components such as myeloperoxidase(MPO),neutrophil elastase(NE)and citrullinated histone H3(CitH3)released by NETs can activate the immune inflammatory cascade,directly damage the vascular endothelium and promote thrombosis.In vascular inflammation,the formation of NETs is regulated by actin,and the released harmful molecules can induce endothelial cell apoptosis and drive the progress of inflammation through oxidative stress.The degradation and clearance of NETs depend on the action of enzymes such as deoxyribonuclease Ⅰ(DNase Ⅰ),and its regulatory mechanisms in atherosclerosis and vascular inflammation remain to be further studied.Based on the above mechanism,NETs-related markers have shown the potential as novel diagnostic and prognostic assessment biomarkers for ASCVD.This article aims to systematically elaborate the core pathological mechanism of NETs driving ASCVD through inflammatory activation,endothelial injury and thrombosis,providing a theoretical basis for targeted intervention.

Objective To investigate the susceptibility and infection characteristics of dengue virus(DENV)in cells derived from diverse tissues of tree shrews and to provide a basis for expanding the repertoire of DENV-permissive cell models in this species.Methods DENV was inoculated at a multiplicity of infection(MOI)of 0.02 into tree shrew skin fibroblasts(TSFs),primary tree shrew renal epithelial cells(pTRECs),tree shrew aortic endothelial cells(TAECs),tree shrew aortic smooth muscle cells(TASMCs),tree shrew hepatocytes(THs),tree shrew corneal stromal cells(TCSCs),tree shrew brain microvascular endothelial cells(TBMECs),and tree shrew retinal microvascular endothelial cells(TRMECs).C6/36,Vero,A549,and BHK-21 cells(commonly used for DENV propagation)were used as positive controls.Over 6 days post-infection,cellular cytopathic effects were monitored at 12-hour intervals using an inverted microscope,viral RNA loads in cell lysates were quantified by real-time fluorescent quantitative PCR to generate proliferation curves,and viral titers were determined by plaque assay.Results Seven types of tree shrew cells,except TRMECs,were susceptible to DENV.Prolonged infection induced pronounced cytopathic effects,including cell rounding,detachment,necrosis,and lysis,across all susceptible cells.The viral RNA loads detected in lysates of pTRECs,TBMECs,TASMCs,TAECs and THs,approached those of positive controls(≥4×107 copies/μL).Infectious progeny viruses were produced by these five cell types,with three(TAECs,3.13×105 PFU/mL;THs,2.03×105 PFU/mL;pTRECs,1.58×105 PFU/mL)exhibiting titers comparable to C6/36(3.85×10 5 PFU/mL)and earlier viral harvests.Conclusion DENV exhibits broad susceptibility to tree shrew cells of multiple tissue origins,with proliferation rates surpassing those of conventional cell lines sourced from other species.TAECs,THs,and pTRECs are particularly suitable for large-scale DENV proliferation,suggesting their potential involvement in in vivo infection.

Objective To investigate the association of spleen volume with the risk of non-alcoholic fatty liver disease(NAFLD)as well as their causal relationship.Methods We included 90 NAFLD cases and 47 healthy controls who had received contrast-enhanced computed tomography(CT)scan of the abdomen at the Second Affiliated Hospital of Xi'an Jiaotong University from November 2022 to November 2023.We conducted three-dimensional reconstruction of the spleen through a deep learning network model using a two-stage coarse-to-fine segmentation approach.We compared the two groups using the two-sample t test or Mann-Whitney U test for continuous data and using the chi-square test for categorical data;evaluated the correlation between spleen volume and liver function indicators through Pearson correlation or Spearman rank correlation analyses;determined the factors influencing the development of NAFLD through multivariable Logistic regression analysis;and further assessed the casual relationship between spleen volume and NAFLD using the inverse variance-weighted two-sample Mendelian randomization(IVW-MR)method.Results Spleen volume was significantly larger in NAFLD cases than in controls(272.93±104.16 vs 204.37±81.20 cm3,P<0.001).The Spearman rank correlation analysis showed that spleen volume was positively correlated with the hepatic steatosis index(rs=0.422,P<0.001)and gamma-glutamyl transferase levels(rs=0.211,P=0.047)in patients with NAFLD.The multivariable Logistic regression analysis indicated that spleen volume was an independent risk factor for the development of NAFLD(odds ratio[OR]=1.01,95%confidence interval[CI]:1.00-1.02,P=0.049).The IVW-MR analysis detected a causal relationship between spleen volume and NAFLD(OR=1.16,95%CI:1.05-1.28,P=0.005).Conclusion Increased spleen volume may be a risk factor for the development and progression of NAFLD.Further studies are still needed to investigate the specific mechanism.

Objective To investigate the effect of medical ozone injection therapy on temporomandibular joint(TMJ)osteoarthritis and its pain in SD rats.Methods Fifity-four rats were randomly assigned according to a random number table into three groups:con-trol group,model group,and medical ozone group,with 18 rats in each group.In the control group,only physiological saline was in-jected during modeling;in the model group,only sodium iodoacetate was injected for modeling;in the medical ozone group,after in-jecting sodium iodoacetate into the joint cavity for modeling for one week,medical ozone was then injected into the joint cavity for inter-vention at a frequency of once a week,totaling 5 times.One week(week 2 after modeling),3 weeks(week 4 after modeling),and 5 weeks(week 6 after modeling)after medical ozone injections,6 rats from each group were euthanized.Mechanical withdrawal thresh-old of rats in each group was assessed before euthanasia,and the expression levels of interleukin-1β(IL-1β)in joint fluid of rats in each group were measured after euthanasia.Gross observation and modified Mankin's scoring were performed on TMJ cartilage of rats in each group after stained with Pelletier score and Safranin O-Fast Green.Results During the same time period,compared to the control group,the model group showed a significant decrease in the mechanical pain threshold of the TMJ in rats at 1 week,3 weeks,and 5 weeks(P＜0.01).The expression levels of IL-1β in the TMJ fluid increased(P＜0.01),and the Pelletier score and modified Mankin's score of TMJ cartilage increased(P＜0.01).In comparison to the model group,the medical ozone group exhibited a significant increase in the mechanical pain threshold of the TMJ in rats after 3 weeks and 5 weeks of medical ozone injections(P＜0.01).The expression levels of IL-1β in the TMJ fluid decreased(P＜0.01),and the Pelletier score and modified Mankin's score of TMJ cartilage decreased(P＜0.01).However,there were no statistically significant difference in the measured parameters in the TMJ cavity after 1 week of medical ozone injection(P＞0.05).Within the medical ozone group,compared to the 1-week treatment,the mechanical pain threshold of the TMJ increased(P＜0.01)and the expression levels of IL-1β in the TMJ fluid decreased at 3 weeks and 5 weeks(P＜0.01).However,there was no statistically significant difference in the Pelletier score and modified Mankin's score of TMJ cartilage(P＞0.05)between different treatment duration.Additionally,there were no statistically signif-icant differences in the mechanical pain threshold of the TMJ,expression levels of IL-1β in the TMJ fluid,Pelletier score,and modi-fied Mankin's score of TMJ cartilage between the medical ozone group at 3 weeks and 5 weeks(P＞0.05).Conclusion Medical ozone treatment for more than 3 weeks can improve temporomandibular joint osteoarthritis and its associated pain in rats.

Objective To establish an immortalized tree shrew corneal stromal cells(CSCs)line and to study its response to virus infection.Methods Primary tree shrew CSCs were isolated and cultured by the tissue block adhesion method.CSCs were then transfected with a lentivirus carrying the SV40T gene and monoclonal cells were selected for passage culture.The characteristics of the CSCs were investigated by morphological observation and compared with 40 generations until the 50 generations or more,immunofluorescence identification of vimentin and SV40T genes,karyotype examination,and cell proliferation curve.The CSCs were infected with herpes simplex virus-1(HSV-1)(McKrae strain),Zika virus(ZIKV,GZ01 strain),Dengue virus typeⅡ,and H1N1(PR8).Results The immortalized tree shrew CSCs after＞50 passages appeared spindle-shaped with good cell morphology and structure compared with 40 generations.Positive immunofluorescence expression of vimentin and SV40T genes.The cell growth curve showed that the cells were in logarithmic-phase growth on days 4～5 and grew vigorously.The number of chromosomes in the primary cells was stable at 62,while immortalized CSCs had 64 chromosomes at P21 and P56.The virus titer results showed that the immortalized tree shrew CSCs were sensitive to HSV-1(McKrae strain),ZIKV(GZ01 strain),Dengue virus typeⅡ,and H1N1(PR8),with virus titers of 1.32×105,5.62×106,2.69×107,and 7.76×104 CCID50/mL,respectively.Conclusions The immortalized tree shrew CSCs were established successfully,suggesting that this cell line is suitable for studies of the mechanisms of HSV,ZIKV,Dengue virus,and influenza A virus infection in relation to corneal diseases and antiviral drugs.