BACKGROUND:Spleen has the functions of blood storage,hematopoiesis,and immunity.With the increase of age,the structural degeneration and functional decline of spleen lead to the impairment of immune system function,thus accelerating the aging process of the body.The treatment of spleen aging in tree shrews with highly active umbilical cord mesenchymal stem cells has not been reported. OBJECTIVE:To explore the intervention effect and mechanism of highly active umbilical cord mesenchymal stem cells on spleen aging in tree shrews. METHODS:Highly active umbilical cord mesenchymal stem cells were isolated,cultured,and obtained from the umbilical cord tissue of newborn tree shrews by caesarean section.The differentiation abilities of adipogenesis,osteogenesis,and chondrogenesis were detected by three-line differentiation kit.Cell cycle and surface markers were detected by flow cytometry.The second generation of highly active umbilical cord mesenchymal stem cells were transfected with Genechem Green Fluorescent Protein with infection complex values of 100,120,140,160,180,and 200,respectively,to screen the best transfection conditions.After transfection,the fourth generation of highly active umbilical cord mesenchymal stem cells was injected into the tail vein of tree shrews in the elderly treatment group.The young control group and the aged model group were not given special treatment.After 4 months of treatment,the spleen tissue was taken and the structure of the spleen was observed by hematoxylin-eosin staining.β-Galactosidase staining was used to detect the activity of aging-related galactosidase.Immunohistochemical staining was used to detect the expression levels of p21 and p53 proteins.Ki67 and PCNA immunofluorescence staining was used to detect cell proliferation activity.Immunofluorescence staining was used to detect the expression levels of spleen autophagy protein molecules Beclin 1 and APG5L/ATG5.Reactive oxygen species fluorescence staining was used to detect the content of reactive oxygen species in spleen tissue.CD3 immunofluorescence staining was used to detect the change of the proportion of total T lymphocytes.The secretion levels of interleukin 1β and transforming growth factor β1 in spleen were detected by enzyme linked immunosorbent assay.The distribution of highly active umbilical cord mesenchymal stem cells labeled with green fluorescent protein in spleen tissue was observed by DAPI double staining of nucleus. RESULTS AND CONCLUSION:(1)Highly active umbilical cord mesenchymal stem cells grew in a short spindle shape with fish-like growth,with a large proportion of G0/G1 phase,and had the potential to differentiate into adipogenesis,osteogenesis,and chondrogenesis.(2)Multiplicity of infection=140 and transfection for 72 hours were the best conditions for labeling tree shrews highly active umbilical cord mesenchymal stem cells with Genechem Green Fluorescent Protein.(3)Compared with the aged model group,in the aged treatment group,the spleen tissue cells of tree shrews were arranged closely,and the area of white pulp was increased(P<0.01);the boundary between red pulp and white pulp was clear;the proportion of germinal centers did not show statistically significant difference(P>0.05).The activity level of galactosidase related to spleen tissue aging was decreased(P<0.001),and the expression levels of aging protein molecules p21 and p53 were down-regulated(P<0.001).The expression levels of proliferation-related molecules Ki67 and PCNA were up-regulated(P<0.001,P<0.05);expression levels of autophagy-related molecules Beclin 1 and APG5L/ATG5 were up-regulated(P<0.001),and the content of reactive oxygen species decreased(P<0.001),and the proportion of CD3+T cells increased(P<0.05).The secretion level of interleukin 1β in the aging-related secretion phenotype decreased(P<0.001);no significant difference was found in transforming growth factor β1 level(P>0.05).Compared with the young control group,the above indexes were significantly different in the elderly treatment group(P<0.05).(4)Green fluorescent cells labeled with green fluorescent protein were observed in spleen tissue of tree shrews the elderly treatment group by frozen tissue section observation.The results show that intravenous infusion of highly active umbilical cord mesenchymal stem cells can migrate to spleen tissue,inhibit the production of reactive oxygen species,down-regulate the expression of aging-related proteins,induce autophagy,promote cell proliferation,reduce chronic inflammation,and then improve the structure and function of spleen tissue.

Objective To investigate the adverse pregnancy outcomes of fetuses with increased nuchal translucency(NT).Methods A total of 376 pregnant women at the Third Affiliated Hospital of Zhengzhou University from May 2023 to January 2024 were selected as research subjects,who had a diagnosis of fetal NT ≥ the 95th percentile and complete pregnancy outcomes for singleton pregnancies.The fetuses were divided into simple thickening group(n=320)and thickening with structural abnormalities group(n=56)based on NT ultrasound results.The interventional prenatal diagnosis outcomes and pregnancy outcomes of two groups were compared.Results The rate of chromosomal abnormalities and the incidence of adverse pregnancy outcomes were higher in thickening with structural abnormalities group compared to simple thickening group with statistically significant differences(P＜0.05).The overall incidence of adverse pregnancy outcomes in NT thickened fetuses was 31.65％,but after excluding chromosomal abnormalities and structural malformations,the good pregnancy outcome rate in NT thickened fetuses was 98.09％.Conclusion NT thickening is associated with adverse pregnancy outcomes in fetuses,and the risk of poor fetal outcome is further increased when NT thickening combined with structural abnormalities in early pregnancy,but the pregnancy outcome is better in fetuses with NT thickening after excluding chromosomal abnormalities and structural malformations.

Objective To investigate the adverse pregnancy outcomes of fetuses with increased nuchal translucency(NT).Methods A total of 376 pregnant women at the Third Affiliated Hospital of Zhengzhou University from May 2023 to January 2024 were selected as research subjects,who had a diagnosis of fetal NT ≥ the 95th percentile and complete pregnancy outcomes for singleton pregnancies.The fetuses were divided into simple thickening group(n=320)and thickening with structural abnormalities group(n=56)based on NT ultrasound results.The interventional prenatal diagnosis outcomes and pregnancy outcomes of two groups were compared.Results The rate of chromosomal abnormalities and the incidence of adverse pregnancy outcomes were higher in thickening with structural abnormalities group compared to simple thickening group with statistically significant differences(P＜0.05).The overall incidence of adverse pregnancy outcomes in NT thickened fetuses was 31.65％,but after excluding chromosomal abnormalities and structural malformations,the good pregnancy outcome rate in NT thickened fetuses was 98.09％.Conclusion NT thickening is associated with adverse pregnancy outcomes in fetuses,and the risk of poor fetal outcome is further increased when NT thickening combined with structural abnormalities in early pregnancy,but the pregnancy outcome is better in fetuses with NT thickening after excluding chromosomal abnormalities and structural malformations.

Objective To screen the risk factors for hypoglycemia in adult intensive care unit(ICU)patients,construct a risk prediction model,and validate its predictive effect.Methods A retrospective study was conducted on adult critically ill patients admitted to the general ICU of Hospital of Chengdu University of Traditional Chinese Medicine from December 2023 to September 2024.Patients admitted from December 2023 to June 2024 served as the modeling group,and those from July to September 2024 as the validation group.A total of 928 patients were included,with 650 in the modeling group and 278 in the validation group.After literature review and expert consultation,27 potential risk factors for hypoglycemia in ICU patients were initially screened,and data were collected including general information[gender,age,acute physiology and chronic health evaluation Ⅱ(APACHEⅡ)score,sequential organ failure assessment(SOFA)score,nutrition risk in critically ill(NUTRIC)score,mechanical ventilation status,hemodialysis status,enteral nutrition status],disease data(sepsis,liver disease history,kidney disease history,diabetes history,hypoglycemia history),blood glucose-related indicators[mean blood glucose,blood glucose coefficient of variation,insulin dosage,intravenous insulin titration use,inotropic drug use,insulin secretagogues(Sulfonylureas and Glinides),and combined use of hypoglycemic drugs(two or more)],and laboratory indicators[serum creatinine(SCr),blood urea nitrogen(BUN),serum albumin(Alb),alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBil),glomerular filtration rate(GFR)].The patients were divided into a hypoglycemia group and a non-hypoglycemia group based on the occurrence of hypoglycemia.Univariate analysis and binary Logistic regression analysis were used to identify influencing factors of hypoglycemia in adult ICU patients,and a nomogram prediction model was constructed.The area under the receiver operator characteristic curve(AUC)and calibration curves were employed to evaluate the discrimination and calibration of the model.Results The modeling cohort included 552 non-hypoglycemic patients and 98 hypoglycemic patients,with an ICU hypoglycemia incidence rate of 15.1％.Compared with the hypoglycemia group,the non-hypoglycemia group showed significantly lower proportions of patients with renal disease history,diabetes history,hypoglycemia history,undergoing hemodialysis,using intravenous insulin titration,and combined use of hypoglycemic drugs,as well as lower blood glucose coefficient of variation,lower APACHEⅡ scores,and significantly elevated GFR(all P<0.05).Binary Logistic regression analysis was performed using the 9 variables with statistically significant differences in univariate analysis as independent variables and hypoglycemia occurrence as the dependent variable.The results indicated that a history of diabetes,a history of hypoglycemia,APACHEⅡ score,GFR,blood glucose coefficient of variation,and combined use of hypoglycemic drugs were independent risk factors for hypoglycemia in ICU patients[odds ratios(OR)were 1.761,2.095,1.048,0.990,1.029,and 1.975,respectively,and 95％confidence intervals(95％CI)were 1.052-2.949,1.220-3.600,1.022-1.074,0.982-0.997,1.013-1.046,and 1.145-3.408,respectively.The corresponding Pvalues were 0.031,0.007,0.000,0.009,<0.001,0.014].A nomogram prediction model for hypoglycemia in ICU patients was constructed using six independent predictors selected through binary logistic regression analysis.The ROC curve AUC for the modeling group was 0.884(95％CI 0.826-0.941,P=0.250),with a maximum Youden index of 0.713,sensitivity of 92.1％,and specificity of 79.2％.The validation cohort included 38 patients with hypoglycemia and 240 patients without hypoglycemia.Compared with the hypoglycemia group,the non-hypoglycemia group showed significantly lower proportions of patients with a history of diabetes,a history of hypoglycemia,and combined use of hypoglycemic drugs,as well as lower APACHEⅡ scores and lower blood glucose coefficient of variation,with significantly increased GFR(all P<0.05).The ROC curve AUC for the validation cohort was 0.803(95％CI was 0.757-0.849,P=0.138),indicating high discriminatory ability.The predicted probability at the diagnostic cutoff point was P=0.138.The model's diagnostic threshold for predicted probability was P=0.138,while the optimal cut-off value based on the Youden index was 0.513,yielding a sensitivity of 76.5％and specificity of 74.8％,indicating predictive value for hypoglycemia in adult ICU patients.The mean absolute error(MAE)results for the modeling group and validation group were<0.05.The calibration curves of both the modeling and validation groups showed close alignment with the ideal curve,indicating excellent calibration performance of the model.Conclusion The constructed hypoglycemia risk prediction model for adult ICU patients has good predictive performance,which can quickly identify high-risk populations of hypoglycemia in ICU and provide reference for clinical preventive nursing.

Aim To explore the impact and mechanism of proprotein convertase subtilisin kexin 9(PCSK9)on the progression of abdominal aortic aneurysm(AAA).Methods 6～8 week old ApoE-/-mice were selected to estab-lish the AAA model.Angiotensin Ⅱ(Ang Ⅱ)was continuously infused through subcutaneous implantation of a micro-os-motic pump.The mice were fed with high-fat diet and killed after 28 days.The expression of PCSK9 in abdominal aor-tic smooth muscle cells was detected by immunohistochemistry and immunofluorescence in normal abdominal aortic blood vessels and AAA samples in human and mice.Primary cultured murine vascular smooth muscle cells(mVSMC)of C57BL/6 mice were treated with different concentrations of AngⅡ for 24 h,and the expression of PCSK9 mRNA and pro-tein was detected.PCSK9 overexpression and knockdown cell models were established,and mitochondrial reactive oxygen species(mtROS),mitochondrial membrane potential(MMP),mitochondrial permeability transition pore(MPTP)open-ing,and Z-DNA binding protein 1(ZBP1)protein expression were detected.Bioinformatics was used to analyze the dif-ferential expression of multiple single-cell sequencing datasets to obtain the key differentially expressed genes,and to study their expression and role in AAA.Results Immunohistochemistry and immunofluorescence results showed that PCSK9 expression in human and mouse AAA increased(P＜0.01),and co-localized with smooth muscle.Ang Ⅱ promoted PCSK9 expression in mVSMC in a concentration-dependent manner,the 2.0 μmol/L Ang Ⅱ group showed a 2.9-fold and 1.1-fold increase in the expression of PCSK9 mRNA and protein,respectively(P＜0.01),with the most significant effect observed.After successfully constructing PCSK9 overexpression and PCSK9 interference mVSMC models,PCSK9 overex-pression led to an increase in intracellular mtROS,a decrease in MMP,an increase in MPTP opening,and a decrease in cellular activity(P＜0.01);PCSK9 knockdown could reduce Ang Ⅱ induced increase in mtROS,decrease in MMP and MPTP opening;compared with the siNC+Ang Ⅱ group,the siPCSK9+Ang Ⅱ group showed a decrease in mtROS and an in-crease in the fluorescence brightness of MMP and MPTP(P＜0.05).Bioinformatics analysis revealed that ZBP1 was a core differentially expressed gene in AAA.Immunohistochemistry and immunofluorescence results showed that ZBP1 ex-pression in human and mouse AAA tissues increased,and co-localized with smooth muscle.Western blot results showed that PCSK9 overexpression or treatment with 2.0 μmol/L Ang Ⅱ could increase ZBP1 protein expression(P＜0.01),while PCSK9 knockdown could alleviate the increased ZBP1 expression caused by AngⅡ(P＜0.05).Conclusion PCSK9 may induce mitochondrial damage in smooth muscle cells,activate downstream molecule ZBP1 to cause cell damage,and promote the development of AAA.

Financial incentives， as an intervention in behavioral therapy， have been proven to be beneficial in many health behaviors， such as drug addiction treatment， long-term antiviral therapy， and the prevention and treatment of chronic diseases. At present， typical financial incentive schemes mainly include commitment contracts and lottery-based models. Key factors in the design of such interventions include the adaptability of behavioral economic principles and medication behavior， the effectiveness and cost-effectiveness of design scheme， and the acceptability of intervention population. Although financial incentives have accumulated rich experience in health behavior interventions abroad， research on medication adherence remains limited， with existing intervention scheme design and mechanism constraints， and their effectiveness potentially varying across different cultural and social environments. China should explore practical models of financial incentives to enhance medication adherence, considering patients’ medication patterns and socio-cultural factors.

Aim To explore the impact and mechanism of proprotein convertase subtilisin kexin 9(PCSK9)on the progression of abdominal aortic aneurysm(AAA).Methods 6～8 week old ApoE-/-mice were selected to estab-lish the AAA model.Angiotensin Ⅱ(Ang Ⅱ)was continuously infused through subcutaneous implantation of a micro-os-motic pump.The mice were fed with high-fat diet and killed after 28 days.The expression of PCSK9 in abdominal aor-tic smooth muscle cells was detected by immunohistochemistry and immunofluorescence in normal abdominal aortic blood vessels and AAA samples in human and mice.Primary cultured murine vascular smooth muscle cells(mVSMC)of C57BL/6 mice were treated with different concentrations of AngⅡ for 24 h,and the expression of PCSK9 mRNA and pro-tein was detected.PCSK9 overexpression and knockdown cell models were established,and mitochondrial reactive oxygen species(mtROS),mitochondrial membrane potential(MMP),mitochondrial permeability transition pore(MPTP)open-ing,and Z-DNA binding protein 1(ZBP1)protein expression were detected.Bioinformatics was used to analyze the dif-ferential expression of multiple single-cell sequencing datasets to obtain the key differentially expressed genes,and to study their expression and role in AAA.Results Immunohistochemistry and immunofluorescence results showed that PCSK9 expression in human and mouse AAA increased(P＜0.01),and co-localized with smooth muscle.Ang Ⅱ promoted PCSK9 expression in mVSMC in a concentration-dependent manner,the 2.0 μmol/L Ang Ⅱ group showed a 2.9-fold and 1.1-fold increase in the expression of PCSK9 mRNA and protein,respectively(P＜0.01),with the most significant effect observed.After successfully constructing PCSK9 overexpression and PCSK9 interference mVSMC models,PCSK9 overex-pression led to an increase in intracellular mtROS,a decrease in MMP,an increase in MPTP opening,and a decrease in cellular activity(P＜0.01);PCSK9 knockdown could reduce Ang Ⅱ induced increase in mtROS,decrease in MMP and MPTP opening;compared with the siNC+Ang Ⅱ group,the siPCSK9+Ang Ⅱ group showed a decrease in mtROS and an in-crease in the fluorescence brightness of MMP and MPTP(P＜0.05).Bioinformatics analysis revealed that ZBP1 was a core differentially expressed gene in AAA.Immunohistochemistry and immunofluorescence results showed that ZBP1 ex-pression in human and mouse AAA tissues increased,and co-localized with smooth muscle.Western blot results showed that PCSK9 overexpression or treatment with 2.0 μmol/L Ang Ⅱ could increase ZBP1 protein expression(P＜0.01),while PCSK9 knockdown could alleviate the increased ZBP1 expression caused by AngⅡ(P＜0.05).Conclusion PCSK9 may induce mitochondrial damage in smooth muscle cells,activate downstream molecule ZBP1 to cause cell damage,and promote the development of AAA.

Objective To screen the risk factors for hypoglycemia in adult intensive care unit(ICU)patients,construct a risk prediction model,and validate its predictive effect.Methods A retrospective study was conducted on adult critically ill patients admitted to the general ICU of Hospital of Chengdu University of Traditional Chinese Medicine from December 2023 to September 2024.Patients admitted from December 2023 to June 2024 served as the modeling group,and those from July to September 2024 as the validation group.A total of 928 patients were included,with 650 in the modeling group and 278 in the validation group.After literature review and expert consultation,27 potential risk factors for hypoglycemia in ICU patients were initially screened,and data were collected including general information[gender,age,acute physiology and chronic health evaluation Ⅱ(APACHEⅡ)score,sequential organ failure assessment(SOFA)score,nutrition risk in critically ill(NUTRIC)score,mechanical ventilation status,hemodialysis status,enteral nutrition status],disease data(sepsis,liver disease history,kidney disease history,diabetes history,hypoglycemia history),blood glucose-related indicators[mean blood glucose,blood glucose coefficient of variation,insulin dosage,intravenous insulin titration use,inotropic drug use,insulin secretagogues(Sulfonylureas and Glinides),and combined use of hypoglycemic drugs(two or more)],and laboratory indicators[serum creatinine(SCr),blood urea nitrogen(BUN),serum albumin(Alb),alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBil),glomerular filtration rate(GFR)].The patients were divided into a hypoglycemia group and a non-hypoglycemia group based on the occurrence of hypoglycemia.Univariate analysis and binary Logistic regression analysis were used to identify influencing factors of hypoglycemia in adult ICU patients,and a nomogram prediction model was constructed.The area under the receiver operator characteristic curve(AUC)and calibration curves were employed to evaluate the discrimination and calibration of the model.Results The modeling cohort included 552 non-hypoglycemic patients and 98 hypoglycemic patients,with an ICU hypoglycemia incidence rate of 15.1％.Compared with the hypoglycemia group,the non-hypoglycemia group showed significantly lower proportions of patients with renal disease history,diabetes history,hypoglycemia history,undergoing hemodialysis,using intravenous insulin titration,and combined use of hypoglycemic drugs,as well as lower blood glucose coefficient of variation,lower APACHEⅡ scores,and significantly elevated GFR(all P<0.05).Binary Logistic regression analysis was performed using the 9 variables with statistically significant differences in univariate analysis as independent variables and hypoglycemia occurrence as the dependent variable.The results indicated that a history of diabetes,a history of hypoglycemia,APACHEⅡ score,GFR,blood glucose coefficient of variation,and combined use of hypoglycemic drugs were independent risk factors for hypoglycemia in ICU patients[odds ratios(OR)were 1.761,2.095,1.048,0.990,1.029,and 1.975,respectively,and 95％confidence intervals(95％CI)were 1.052-2.949,1.220-3.600,1.022-1.074,0.982-0.997,1.013-1.046,and 1.145-3.408,respectively.The corresponding Pvalues were 0.031,0.007,0.000,0.009,<0.001,0.014].A nomogram prediction model for hypoglycemia in ICU patients was constructed using six independent predictors selected through binary logistic regression analysis.The ROC curve AUC for the modeling group was 0.884(95％CI 0.826-0.941,P=0.250),with a maximum Youden index of 0.713,sensitivity of 92.1％,and specificity of 79.2％.The validation cohort included 38 patients with hypoglycemia and 240 patients without hypoglycemia.Compared with the hypoglycemia group,the non-hypoglycemia group showed significantly lower proportions of patients with a history of diabetes,a history of hypoglycemia,and combined use of hypoglycemic drugs,as well as lower APACHEⅡ scores and lower blood glucose coefficient of variation,with significantly increased GFR(all P<0.05).The ROC curve AUC for the validation cohort was 0.803(95％CI was 0.757-0.849,P=0.138),indicating high discriminatory ability.The predicted probability at the diagnostic cutoff point was P=0.138.The model's diagnostic threshold for predicted probability was P=0.138,while the optimal cut-off value based on the Youden index was 0.513,yielding a sensitivity of 76.5％and specificity of 74.8％,indicating predictive value for hypoglycemia in adult ICU patients.The mean absolute error(MAE)results for the modeling group and validation group were<0.05.The calibration curves of both the modeling and validation groups showed close alignment with the ideal curve,indicating excellent calibration performance of the model.Conclusion The constructed hypoglycemia risk prediction model for adult ICU patients has good predictive performance,which can quickly identify high-risk populations of hypoglycemia in ICU and provide reference for clinical preventive nursing.

Purpose/Significance To analyze the dynamic changes,influencing factors and weak links of e-health literacy of middle-aged and elderly people,so as to provide references for improving e-health literacy of middle-aged and elderly people.Method/Process A questionnaire survey is conducted among 505 middle-aged and elderly people in Guangzhou,and the results are analyzed by using SPSS 24.0.Result/Conclusion The average score of e-health literacy of middle-aged and elderly people is in the middle level;The weak link of e-health literacy is the lack of health information interaction and evaluation capacity.Individual factors are the primary factors that affect e-health literacy of middle-aged and elderly people.It is suggested to increase policy and financial support,carry out"the younger population helps the aging population"mode,and elderly-oriented transformation of digital health Apps.

Objective To investigate the effect of cisplatin treatment on the transcriptional level of human liver cancer cells by conducting transcriptome sequencing analysis after treating human liver cancer cell lines with differ-ent concentrations of cisplatin(CDDP).Methods Liver cancer cell lines HepG2 and Huh7 were incubated with cisplatin at different final concentrations of 0,20,50,100 and 200 μmol/L.After 12 hours,cell viability,immuno-fluorescence and RNA-sequencing(RNA-seq)were performed.Differential gene expression analysis(DEG),KEGG pathway analysis,and protein-protein interaction network analysis were conducted.Results Cisplatin de-creased cell viability and increased DNA damage in HepG2,Huh7 cells.Among the genes regulated after cisplatin treatment at different concentrations,59 genes were commonly up-regulated in both HepG2 and Huh7 cells,while 81 genes were commonly down-regulated.The commonly upregulated genes were mainly enriched in cancer initiation and progression pathways.The 81 commonly down-regulated genes were mainly enriched in Rap1 signaling pathway,Ras signaling pathway,signaling pathways regulating pluripotency of stem cells,axon guidance,and cell adhesion-related pathways.Survival analysis of key nodes in the protein-protein interaction network of commonly up-regulated and downregulated genes revealed a significant correlation between high expression of Jun proto-oncogene,AP-1 transcription factor subunit(JUN)and prolonged patient survival and a significant correlation between low ex-pression of growth arrest and DNA damage inducible alpha(GADD45A)and prolonged patient survival.Conclu-sions The study revealed common transcriptional changes in liver cancer cells under cisplatin treatment.Differential expression of JUN and GADD45A is a potential core mechanism to explain drug resistance.This conclusion provides some important prognostic indicators for clinical treatment.