Objective To explore the association between triglyceride glucose-body mass index (TyG-BMI) and the risk of cardiovascular and cerebrovascular diseases in middle-aged and elderly (aged≥45 years) people, and to provide a scientific basis for the prevention and control of cardiovascular and cerebrovascular diseases. Methods A total of 859 patients with cardiovascular and cerebrovascular diseases who were hospitalized in the Department of General Medicine of Heping Hospital affiliated to Changzhi Medical College from January 2020 to November 2024 were selected as the observation group, and 859 patients with non-cardiovascular and cerebrovascular diseases hospitalized during the same period were selected and matched 1:1 by age (±2 years) and gender as the control group. The TyG-BMI index was compared between the two groups，and the TyG-BMI was divided into three groups according to the control group quartile: low-level group (Q1:< P25), medium-level group (Q2:P25-P75), and high-level group (Q3:>P75). The relationship between the TyG-BMI and the risk of cardiovascular and cerebrovascular diseases was analyzed by conditional logistic regression. Results The TyG-BMI score in the case group was significantly higher than that in the control group (t=14.944，P<0.001), with the most significant difference in the age group of 45-55 years old (t=14.276, P<0.001). Multivariate logistic regression analysis showed that each unit increase in the TyG-BMI index was associated with a 1.406-fold increased risk of cardiovascular and cerebrovascular disease (95% CI: 1.231-2.301). The analysis of different index levels showed that compared with low-level TyG-BMI, the risk of cardiovascular and cerebrovascular diseases was higher at medium or high TyG-BMI. The subgroup analysis showed that the TyG-BMI index only had an effect on cardiovascular disease (OR=1.472, 95%CI: 1.141-3.032). Conclusion The TyG-BMI is positively correlated with the occurrence of cardiovascular and cerebrovascular diseases in middle-aged and elderly people, with a stronger association with cardiovascular disease.

Objective To investigate whether microRNA-21-5p(miR-21)plays a protective role in hyperoxia-induced acute lung injury(HALI)by regulating ferroptosis through the STAT3/P53/SLC7A11 axis.Methods The interaction between STAT3 and P53 was analyzed by co-immunoprecipitation(Co-IP).Fifty 9-week-old male C57BL/6J mice were randomly divided into normoxia(Control)group,HALI group,miR-21 overexpression(miR-21)group,STAT3 inhibitor(HY-13818)group,and ferrostatin-1(Fer-1)group.After the mice from the miR-21 group received miR-21-5p AAV6 or empty vector via tracheal catheter instillation,the animals were then monitored for 3 weeks.The HY-13818 group was intraperitoneally injected with HY-13818(10 mg/kg)3 times weekly for 2 weeks.The Fer-1 group received 0.8 mg/kg ferrostatin-1 via tail vein injection once daily for 2 consecutive days during modeling.The HALI model was established by exposure to>90%oxygen.After 48 h of hyperoxia,blood samples were collected via orbital sampling for RT-PCR analysis of miR-21 expression.Lung tissues were harvested for wet/dry weight ratio and assessment of histopathological changes via HE staining for lung injury score.Activity of superoxide dismutase(SOD)and contents of malondialdehyde(MDA),Fe2?,glutathione(GSH),and reactive oxygen species(ROS)were measured using photocolorimetry,spectrophotometry and fluorometry.Western blotting was used to evaluate the protein expression of STAT3,P53,SLC7A11,and GPX4.Results The results of Co-IP showed that STAT3 could bind to P53.The HALI group exhibited obviously destroyed alveolar structure,disordered arrangement,thickened interval,with a large number of infiltrated neutrophils and collapsed alveoli,and had significantly increased pathological score of lung injury and ratio of lung Wwet/Ddry weight when compared with the Control group(P<0.05).In the miR-21 group,HY-13818 group and Fer-1 group,the severity of lung injury was significantly reduced,and the pathological score of lung injury and the ratio of Wwet/Ddry weight were decreased(P<0.05)when compared with the HALI group.Compared with the control group,the contents of MDA,Fe2+and ROS were increased(P<0.05),the activity of SOD and content of GSH were declined(P<0.05),the protein levels of STAT3 and P53 were increased(P<0.05),and those of SLC7A11 and GPX4 were decreased(P<0.05)in the HALI group.Compared with the HALI group,decreased MDA and ROS levels(P<0.05),enhanced SOD activity,Fe2+and GSH levels(P<0.05),down-regulation of STAT3 and P53(P<0.05)and up-regulation of SLC7A11 and GPX4(P<0.05)were observed in the miR-21 group and HY-13818 group.Conclusion MiR-21 alleviates HALI,which may be related to its inhibition of ferroptosis through the STAT3/P53/SLC7A11 axis.

Objective To investigate the effects of Sini Powder on serum hypothalamic-pituitary-adrenal(HPA)axis-related hormones and inflammatory factors in liver cancer mice with comorbid depression,and to evaluate its effect on depressive behavior and tumor proliferation activity.Methods Forty-eight specific pathogen-free female C57BL/6 mice were randomly assigned to either a blank(n=8)or model group(n=40).The modeling group was subjected to chronic unpredictable mild stress(CUMS)for six weeks.Both groups underwent orthotopically transplanted liver tumor surgery at the end of the fourth week of CUMS treatment.At the end of the sixth week of CUMS treatment,color Doppler ultrasonography was used to observe tumor formation in the orthotopic transplantation liver tumors,and the tail suspension test was used to assess depressive behavior.Non-tumor-bearing and deceased mice were excluded.The remaining model group mice were stratified by tail suspension immobility time and randomly assigned to the following groups:model group(distilled water),Fluoxetine group(5.0 mg/kg),and Sini Powder low-dose,medium-dose,and high-dose groups(5.2,10.4,and 20.8 g/kg,respectively),with six mice per group.The treatments were administered once daily for 21 consecutive days.After treatment,depressive behaviors were assessed using the open field,tail suspension,and forced swimming tests.The proliferation status of the orthotopic liver transplantation tumor was evaluated by measuring the size of the tumor,observing pathological changes in the tumor tissue through hematoxylin and eosin staining,and detecting the positive cell rate of proliferating cell nuclear antigen(Ki-67)in the tumor tissue using immunohistochemistry.The levels of HPA axis-related hormones in serum,such as corticotropin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),corticosterone(CORT),as well as inflammatory factors such as tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-6(IL-6)were measured using an enzyme-linked immunosorbent assay.Western blotting was used to assess mitogen-activated protein kinase(MAPKs)phosphorylation and the expression of nuclear factor-κB(NF-κB),NOD-like receptor family pyrin domain-containing receptor 3(NLRP3),and cysteine aspartic protease-1(Caspase-1)in orthotopic tumors.Results Compared with the blank group,the model group showed reduced total distance traveled in open field test,prolonged immobility times in the tail suspension and forced swimming tests(P<0.05,P<0.01),indicating successful establishment of the liver cancer with comorbid depression mice model.Also,the model group showed increased orthotopic tumor volume(P<0.01),and elevated serum CRH,ACTH,CORT,TNF-α,IL-1β,and IL-6 levels(P<0.01).The phosphorylation of MAPKs in tumor tissues was suppressed(P<0.01),while NF-κB,NLRP3,and Caspase-1 expression levels were downregulated(P<0.01).Compared with the model group,Sini Powder medium-and high-dose groups exhibited increased total distance traveled in the open field test(P<0.05),reduced forced swimming test and prolonged total distance traveled in open field test(P<0.01),while Sini Powder high-dose group showed reduced immobility times in the tail suspension test(P<0.05).Also,Sini Powder low-dose,medium-dose,and high-dose groups showed slower tumor growth,histological changes,including vacuolization and necrosis,decreased Ki-67 positive cell rate(P<0.01),and reduced serum CRH,ACTH,CORT,TNF-α,IL-1β,and IL-6 levels(P<0.05).Additionally,the phosphorylation of MAPKs in tumor tissues was suppressed(P<0.01),and NF-κB,NLRP3,and caspase-1 expression levels were downregulated(P<0.01).Conclusion Sini Powder may alleviate depressive behaviors and suppress tumor proliferation activity in liver cancer mice with comorbid depression by modulating MAPKs activation,inhibiting NF-κB,NLRP3,and Caspase-1 expressions,and reducing serum inflammatory factors and HPA axis-related hormones levels.

Objective To explore the effect of Osthole on neuroinflammation in Alzheimer's disease(AD)by regulating the lactylation of pyruvate kinase M2(PKM2).Methods(1)Animal experiments:18 mice were divided into three groups,namely wild-type(WT)group,APP/PS1 group and APP/PS1+Osthole group.Learning-and memory-related biobehavioral indicators were compared among the three groups.Immunohistochemistry was used to detect the positive expression of Iba1 in brain tissue,enzyme-linked immunosorbent assay(ELISA)was employed to detect the levels of interleukin(IL)-6,tumor necrosis factor(TNF)-α,and IL-1β in brain tissue,and Western Blot was used to detect the protein expression levels of Pan lactylation(Pan-kla)and PKM2 lactylation(PKM2-kla)in brain tissue.(2)Cell experiments:an in vitro AD model was constructed by treated in mouse microglia(BV2 cells)with LPS/Aβ1-42,and followed by treatment with Osthole.Cell viability was detected by methyl thiazolyl tetrazolium(MTT),expression of Iba1(a marker of microglial activation)was detected by Western Blot,nitric oxide(NO)production was assessed by Griess reagent,and levels of IL-6,TNF-α and IL-1β were detected by ELISA.BV2 cell-conditioned medium(CM)was co-cultured with neuroblastoma cells(Na2 cells)to assess the protective effect of Osthole on Na2 cells.(3)Molecular docking was performed between Osthole and PKM2,and experimental verification was conducted.Results In animal experiments,deficits of learning and memory in mice were aggravated in APP/PS1 group compared with that in WT group,which were improved upon treatment with Osthole.Furthermore,the APP/PS1 group mice showed an increase in Iba1 positive cells in brain tissue,an increase in the levels of pro-inflammatory factors IL-6,TNF-α and IL-1β,as well as an increase in the levels of Pan-kla and PKM2-kla compared with the WT group,while the above indexes were inhibited by the Osthole treatment.In cell experiments,Osthole had no significant effect on BV2 cell viability at concentrations up to 100 μmol/L.Treatment with LPS/Aβ1-42 upregulated the expression of Iba1,NO production,and levels of pro-inflammatory factors IL-6,TNF-α,and IL-1β in BV2 cells,while Osthole significantly inhibited the expression of these LPS/Aβ1-42-induced indicators.Meanwhile,Osthole attenuated the damage of BV2-CM on Na2 cells.The molecular docking results indicated a good binding affinity between Osthole and PKM2.Treatment with Osthole can down-regulated the levels of lactate,Pan-kla and PKM2-kla in the AD cell model.Conclusion Osthole can improve the condition of AD and reduce neuroinflammation by inhibiting the lactylation of PKM2.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective:To summarize domestic and foreign transitional readiness assessment tools for adolescents with chronic diseases and analyze the current status of their application in the clinic, so as to provide references for healthcare professionals to select appropriate tools.Methods:China National Knowledge Infrastructure, Wanfang Data, VIP and China Biomedical Literature Database, PubMed, Web of Science, Embase, Medline, CINAHL, Cochrane Library and scholar.google.com were searched for literature related to the assessment tools of transition readiness in adolescents with chronic illnesses from inception to March 29, 2024. Two researchers independently screened the literature and extracted the data.Results:A total of 491 articles were initially retrieved and 24 were included, involving a total of 8 transitional readiness assessment tools for adolescents with chronic diseases, including the Transition Readiness Assessment questionnaire, the University of North Carolina TRxANSITION Scale, Am I ON TRAC for adult care? Questionnaire, Transition Questionnaire, Self-Management and Transition to Adulthood with Rx=Treatment Questionnaire, Good 2 Go Questionnaire, State Assessment Questionnaire for Transition, and Self-assessment Scale of Transition Readiness for Adolescents. The Transition Readiness Assessment questionnaire and Self-Management and Transition to Adulthood with Rx=Treatment Questionnaire have good reliability and validity, comprehensive evaluation, wide applicability, simple use, and are suitable for clinical use.Conclusions:The quality of existing tools for assessing transitional readiness of adolescents with chronic diseases is mixed. The Transition Readiness Assessment questionnaire and Self-Management and Transition to Adulthood with Rx=Treatment Questionnaire are more appropriate options in the clinic, but they still need to be improved. Future studies need to continue to introduce tools to assess transitional readiness for chronic diseases in adolescents and develop localized assessment tools.

The novel coronavirus(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2),classified as a single-stranded RNA virus,replicates and transcribes its genome through the action of an RNA-dependent RNA polymerase,which is itself comprised of numerous non-structural pro-teins(non-structural proteins,Nsps).The present study delineates the development of a detection system founded on a bicistronic reporter plasmid in conjunction with an array of Nsp plasmids,ai-ming to investigate the influence of SARS-CoV-2 Nsps on the virus's replication and expression profiles.Specifically,a bicistronic reporter gene plasmid along with twelve distinct Nsp plasmids(encompassing Nsp3C-Flag,Nsp4,Nsp6-Nsp10,Nsp12-Nsp16)were meticulously constructed via molecular cloning techniques.The successful expression of these Nsps was subsequently confirmed through Western blot analysis.Initially,the activity of the RNA-dependent RNA polymerase was assessed by co-transfection of the reporter plasmid with Nsp12,in the presence of its auxiliary fac-tors Nsp7 and Nsp8,with careful regulation of the co-transfection ratio,culturing temperature,and the timing of activity determination for the triad of Nsp plasmids.The normalized NLuc fluorescein value,in reference to the FLuc fluorescein value of the housekeeping gene,served as a metric for determining the polymerase activity.Building upon this foundation,the co-transfection concentra-tions of Nsp9-16 were fine-tuned,followed by the incremental addition of varying doses of Nsp3C,Nsp4,and Nsp6,to further elucidate the activity of RNA-dependent RNA polymerase(RdRp).The findings indicated that upon transfection with varying ratios of Nsp7,Nsp8,and Nsp12 at a propor-tion of 1∶8∶24,the polymerase activity was markedly elevated compared to the control group,with a statistical significance level(P＜0.001).Furthermore,in the absence of Nsp3,Nsp4,and Nsp6,the inclusion of Nsp10-16 substantially augmented the activity of the RdRp,particularly in scenarios where Nsp9 was not introduced,achieving statistical significance(P＜0.001).In the pres-ence of Nsp3 and Nsp4,the RdRp activity was augmented further upon the addition of Nsp9,reac-hing a level of significance(P＜0.05).The data imply that Nsp9 is capable of enhancing the RdRp activity of SARS-CoV-2 exclusively in the context of Nsp3 and Nsp4 coexistence,suggesting that the stimulatory influence of Nsp9 on viral replication may be contingent upon the formation of double-membrane vesicles.

This study aims to prepare monoclonal antibody to S1 protein of porcine epidemic diar-rhea virus(PEDV).E.coli expression system and affinity chromatography were used to success-fully obtain purified recombinant PEDV S1 protein.After immunizing BALB/c mice,hybridoma technology and indirect ELISA were used to prepare and screen positive hybridoma cells.Finally,ascites antibodies were prepared by in vivo induction method.ELISA results showed that a total of 4 hybridoma cell lines with anti-PEDV S1 monoclonal antibody were screened,and they were named E6,G3,H6 and F2.The supernatant titers of all 4 hybridoma cell lines reached 1∶6 400.The monoclonal antibody H6 with higher antibody titers and more stable antibody secretion was selected for antibody type identification.It was found that monoclonal antibody H6 belongs to the IgG1 subclass and the light chain is the λ chain.The antibody titers that induced mouse ascites were 1∶106 and without cross-reaction with other proteins.Western blot results showed that the monoclonal antibody exhibited specific bands at 38 kDa with the recombinant S1 protein,PEDV QY2016,and PEDV CV777 strains.The IFA results also showed that the monoclonal antibody reacted with cells infected with PEDV QY2016 and PEDV CV777 strains,exhibiting a green fluo-rescent signal.The affinity constant of monoclonal antibody H6 was K=1.75×107 moL/L,indica-ting that the H6 strain had a good affinity and could be used for the development of subsequent di-agnostic antibodies.In summary,this study successfully prepared monoclonal antibodies that can specifically recognize PEDV S1 protein,which can be used for the antigen detection of PEDV and providing important test materials for the research of PEDV detection methods.

Objective To quantitatively evaluate the structure and content of the long-term care insurance(LTCI)policy in China's deeply aging areas.Methods Using the Policy Modeling Consistency(PMC)index model indicator design method,a LTCI policy evalua-tion index system was constructed,consisting of nine primary indicators and 34 secondary indicators.A total of 123 provincial-level LTCI policies issued in deeply aging regions of China between June 1,2014,and October 1,2024 were analyzed.High-frequency word extraction was performed using ROSTCM 6.0,and a social network diagram of LTCI policies was created.The policy structure and content were quantitatively evaluated and ana-lyzed based on the established policy evaluation index system.Results The main content of LTCI policies in deeply aging areas of China covered services,institutions and assessment.The highest policy score was 7.28,and the lowest was 2.20,with an average score of 5.00.There were 25 perfect policies,63 excellent policies,28 good policies and seven qualified policies.In the dimension of policy content,the indexes of five primary indicators of policy evaluation,policy target groups,policy nature,policy perspective and policy tools were 0.60 or more;while the indexes of four primary indicators of policy content,incentives and constraints,policy timeliness,and policy level were 0.50 or less.Conclusion LTCI policies issued in China's deeply aging areas provide comprehensive coverage in aspects such as poli-cy evaluation,policy target groups and policy nature,and need to be improved in policy tool selection and the construction of incentive and constraint mechanisms.

The novel coronavirus(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2),classified as a single-stranded RNA virus,replicates and transcribes its genome through the action of an RNA-dependent RNA polymerase,which is itself comprised of numerous non-structural pro-teins(non-structural proteins,Nsps).The present study delineates the development of a detection system founded on a bicistronic reporter plasmid in conjunction with an array of Nsp plasmids,ai-ming to investigate the influence of SARS-CoV-2 Nsps on the virus's replication and expression profiles.Specifically,a bicistronic reporter gene plasmid along with twelve distinct Nsp plasmids(encompassing Nsp3C-Flag,Nsp4,Nsp6-Nsp10,Nsp12-Nsp16)were meticulously constructed via molecular cloning techniques.The successful expression of these Nsps was subsequently confirmed through Western blot analysis.Initially,the activity of the RNA-dependent RNA polymerase was assessed by co-transfection of the reporter plasmid with Nsp12,in the presence of its auxiliary fac-tors Nsp7 and Nsp8,with careful regulation of the co-transfection ratio,culturing temperature,and the timing of activity determination for the triad of Nsp plasmids.The normalized NLuc fluorescein value,in reference to the FLuc fluorescein value of the housekeeping gene,served as a metric for determining the polymerase activity.Building upon this foundation,the co-transfection concentra-tions of Nsp9-16 were fine-tuned,followed by the incremental addition of varying doses of Nsp3C,Nsp4,and Nsp6,to further elucidate the activity of RNA-dependent RNA polymerase(RdRp).The findings indicated that upon transfection with varying ratios of Nsp7,Nsp8,and Nsp12 at a propor-tion of 1∶8∶24,the polymerase activity was markedly elevated compared to the control group,with a statistical significance level(P＜0.001).Furthermore,in the absence of Nsp3,Nsp4,and Nsp6,the inclusion of Nsp10-16 substantially augmented the activity of the RdRp,particularly in scenarios where Nsp9 was not introduced,achieving statistical significance(P＜0.001).In the pres-ence of Nsp3 and Nsp4,the RdRp activity was augmented further upon the addition of Nsp9,reac-hing a level of significance(P＜0.05).The data imply that Nsp9 is capable of enhancing the RdRp activity of SARS-CoV-2 exclusively in the context of Nsp3 and Nsp4 coexistence,suggesting that the stimulatory influence of Nsp9 on viral replication may be contingent upon the formation of double-membrane vesicles.