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 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.

Objective To systematically evaluate the assessment tools of adolescent chronic disease patients'transition readiness to adults at home and abroad,and to provide references for medical personnel to choose appropriate tools.Methods CNKI,Wan Fang Data,CBM,PubMed,Web of Science,Embase and Cochrane were searched for literature related to the assessment tools of transition readiness in adolescents with chronic illnesses to adulthood from inception to August,2024.Literature screening and data extraction of the tools were extracted independently by 2 researchers.At the same time,the criteria for the selection of health measurement tools based on consensus(COSMIN)systematic evaluation guidelines were used to evaluate the inclusion assessment tools,and recommendations were finally formed.Results A total of 29 articles were included,involving 9 universal and 5 specific assessment tools of adolescent chronic illnesses transition readiness.The Transition Readiness Assessment Questionnaire has satisfactory content validity and internal consistency,and it is recommended as Grade A.The Grade C included the Am I ON TRAC for adult care?Questionnaire and the State Assessment Questionnaire for Transition,Epilepsy-specific Transition Readiness Assessment Questionnaire and the Readiness for Adult Care in Rheumatology,while the others are recommended as Grade B.Conclusion There are a variety of tools to assess the transition readiness of adolescents with chronic diseases to adults,and there are few tools suitable for adolescents with chronic diseases in China,with few specific tools.After comprehensive consideration,the Transition Readiness Assessment Questionnaire can be recommended temporarily.

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 By observing the regulatory effect of Strengthening Spleen and Eliminating Cancer Formula on N6-methyladenosine(m6A)methyltransferase,To explore the effect of Strengthening Spleen and Eliminating Cancer Formula on inhibiting the IRX5 m6A level in colorectal cancer(CRC),the regulatory effect on N6-methyladenosine(m6A)methyltransferase was observed.Methods Clinically,m6A hypermethylated genes in colorectal cancer was analyzed by m6A sequencing of pathological tissues from five CRC patients after radical surgery,looking for protein detection indexes for validation.23 BALB/c nude mice were selected and injected with HCT116 cells to establish a nude-mouse transplantation model of human colorectal cancer.They were divided into Model group,Western medicine group(5-fluorouracil group),Chinese medicine group(Jianpi Xiaoai Formula low-dose group,Jianpi Xiaoai Formula high-dose group),with 6 rats in each group,5 rats in control group.The tumor volume of all groups was compared.The overall methylation level of m6A was detected by colorimetric method.The protein expression levels of METTL3,METTL14,and WTAP,in tumor were detected by Western blot.The SRAMP website was used to predict the m6A sites of IRX5.HCT116 cells were treated with oe-NC,oe-METTL3,sh-NC,and sh-METTL3.The expression of IRX5 protein was detected by Western blot.HCT116 cell line was treated with Jianpi Xiaoai Formula drug-containing serum,and transfected with oe-METTL3 and oe-IRX5.The group was set as followed:control group,Jianpi Xiaoai Formula drug-containing serum group,Jianpi Xiaoai Formula drug-containing serum group+oe-NC,Jianpi Xiaoai Formula drug-containing serum group+oe-METTL3,Jianpi Xiaoai Formula drug-containing serum group+oe-IRX5,cell cloning experiment and Transwell experiment were performed to detect cell proliferation,migration and invasion ability of each group.The protein expression levels of METTL3 and IRX5 were detected by Western blot.Results The results of m6A sequencing of genes showed that the m6A methylation level increased in patients with CRC,and the m6A methylation levels of SOX1 and IRX5 were significantly elevated.Compared with the model group,the tumor volume of Jianpi Xiaoyou Formula high-dose group,low-dose group and 5-Fu group decreased significantly(P<0.01),and the tumor inhibition effect was more obvious with the increase of Jianpi Xiaoai Formula concentration(P<0.01).The methylation level of m6A in Jianpi Xiaoai Formula high dose group,low dose group and 5-Fu group decreased significantly(P<0.01).The SRAMP website predicted that IRX5 contained multiple m6A sites.Overexpression of METTL3 promoted the expression of IRX5 protein(P<0.001),while knockdown of METTL3 inhibited the expression of IRX5 protein(P<0.001).The drug-containing serum of Jianpi Xiaoai Formula could inhibit the protein expression of METTL3 and IRX5(P<0.05)and inhibit the proliferation,migration and invasion of HCT116(P<0.01).Overexpression of METTL3 and IRX5 reversed the inhibitory effect of Jianpi Xiaoai Formula on HCT116 evil phenotype(P<0.01).Conclusion Jianpi Xiaoai Formula may inhibit METTL3 expression mediated IRX5 low expression to inhibit the progression of colorectal cancer.

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.

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.

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.

Objective By observing the regulatory effect of Strengthening Spleen and Eliminating Cancer Formula on N6-methyladenosine(m6A)methyltransferase,To explore the effect of Strengthening Spleen and Eliminating Cancer Formula on inhibiting the IRX5 m6A level in colorectal cancer(CRC),the regulatory effect on N6-methyladenosine(m6A)methyltransferase was observed.Methods Clinically,m6A hypermethylated genes in colorectal cancer was analyzed by m6A sequencing of pathological tissues from five CRC patients after radical surgery,looking for protein detection indexes for validation.23 BALB/c nude mice were selected and injected with HCT116 cells to establish a nude-mouse transplantation model of human colorectal cancer.They were divided into Model group,Western medicine group(5-fluorouracil group),Chinese medicine group(Jianpi Xiaoai Formula low-dose group,Jianpi Xiaoai Formula high-dose group),with 6 rats in each group,5 rats in control group.The tumor volume of all groups was compared.The overall methylation level of m6A was detected by colorimetric method.The protein expression levels of METTL3,METTL14,and WTAP,in tumor were detected by Western blot.The SRAMP website was used to predict the m6A sites of IRX5.HCT116 cells were treated with oe-NC,oe-METTL3,sh-NC,and sh-METTL3.The expression of IRX5 protein was detected by Western blot.HCT116 cell line was treated with Jianpi Xiaoai Formula drug-containing serum,and transfected with oe-METTL3 and oe-IRX5.The group was set as followed:control group,Jianpi Xiaoai Formula drug-containing serum group,Jianpi Xiaoai Formula drug-containing serum group+oe-NC,Jianpi Xiaoai Formula drug-containing serum group+oe-METTL3,Jianpi Xiaoai Formula drug-containing serum group+oe-IRX5,cell cloning experiment and Transwell experiment were performed to detect cell proliferation,migration and invasion ability of each group.The protein expression levels of METTL3 and IRX5 were detected by Western blot.Results The results of m6A sequencing of genes showed that the m6A methylation level increased in patients with CRC,and the m6A methylation levels of SOX1 and IRX5 were significantly elevated.Compared with the model group,the tumor volume of Jianpi Xiaoyou Formula high-dose group,low-dose group and 5-Fu group decreased significantly(P<0.01),and the tumor inhibition effect was more obvious with the increase of Jianpi Xiaoai Formula concentration(P<0.01).The methylation level of m6A in Jianpi Xiaoai Formula high dose group,low dose group and 5-Fu group decreased significantly(P<0.01).The SRAMP website predicted that IRX5 contained multiple m6A sites.Overexpression of METTL3 promoted the expression of IRX5 protein(P<0.001),while knockdown of METTL3 inhibited the expression of IRX5 protein(P<0.001).The drug-containing serum of Jianpi Xiaoai Formula could inhibit the protein expression of METTL3 and IRX5(P<0.05)and inhibit the proliferation,migration and invasion of HCT116(P<0.01).Overexpression of METTL3 and IRX5 reversed the inhibitory effect of Jianpi Xiaoai Formula on HCT116 evil phenotype(P<0.01).Conclusion Jianpi Xiaoai Formula may inhibit METTL3 expression mediated IRX5 low expression to inhibit the progression of colorectal cancer.