Objective To explore the effects and mechanisms of Wendan Decoction(WDT)on anxiety-like and fear-like behaviors,as well as sleep,in the rat model of insomnia due to heart-gallbladder qi deficiency. Methods SD male rats were randomly allocated into blank,model,Anshen Dingzhi pills(0.54 g/kg),low-,medium-,and high-dose(8.7,17.4,and 34.8 g/kg)WDT groups.Plantar electric stimulation combined with forced exercise was used for modeling,and the rats were administrated with corresponding agents by gavage for 7 consecutive days after the modeling.Behavioral tests were performed,and the c-Fos protein level,neuron firing rate,and hippocampal neurotransmitter content in brain regions were determined. Results Compared with the model group,medium-dose WDT shortened the avoidance time,freezing time,and sleep latency(P<0.001,P<0.001,P=0.010).Furthermore,medium-dose WDT attenuated the elevated c-Fos expression in emotion-related brain regions,including the prelimbic cortex(P<0.001),infralimbic cortex(P<0.001),basolateral amygdala(P<0.001),central amygdala(P<0.001),ventral hippocampus(P=0.004),periaqueductal gray(P<0.001),and dorsal raphe nucleus(P<0.001),as well as the increased firing rate of ventral hippocampal interneurons(P=0.005).In addition,it reduced the hippocampal glutamate content and the glutamate/gamma-aminobutyric acid ratio(P<0.001,P=0.013). Conclusions WDT can reduce the excitatory drive of glutamic acid to interneurons by inhibiting the release of glutamic acid from pyramidal neurons of ventral hippocampus,thus playing a role in improving sleep and mitigating the anxiety-like,fear-like,and depression-like behaviors in the rat model of insomnia due to heart-gallbladder qi deficiency.

Objective To identify key genes of lipid metabolism in diabetic nephropathy(DN) through machine learning models and animal model validation. Methods The limma R package was used for differential gene expression analysis on 69 samples from two transcriptome datasets of the Gene Expression Omnibus and 2 184 differentially expressed genes were identified.Subsequently,we adopted undifferentiated consensus clustering to classify DN samples into two specific subtypes.At the same time,we performed weighted gene co-expression network analysis to mine the gene modules significantly associated with DN.In addition,using least absolute shrinkage and selection operator,support vector machine-recursive feature elimination,and random forest machine learning techniques,combined with protein-protein interaction network analysis,we screened out three core genes.Finally,we constructed a mouse model of type 2 diabetes mellitus to verify the effectiveness of the expression of these key genes. Results Three core genes,APOO,ALDH7A1,and ALB,were predicted as potential biomarkers of lipid metabolism in DN,and their expression levels were downregulated in DN.Through experimental validation in a diabetic mouse model,we confirmed the altered expression of APOO,ALDH7A1,and ALB in DN,which supported their potential as diagnostic markers. Conclusions Our findings suggest that APOO,ALDH7A1,and ALB are new diagnostic markers associated with lipid metabolism in DN,which provides new perspectives for understanding the molecular mechanisms of lipid metabolism in DN.

Objective To explore the association between dietary chili intake and cognitive function in the elderly population and the modifying effect of body mass index. Methods On the basis of the baseline survey data of the West China Elderly Health Cohort,a cross-sectional design was adopted,and 1 961 older adults aged 60 and above who had completed a dietary survey were included in this study.Multivariate linear regression and multivariate Logistic regression models were used to explore the associations of chili intake with cognitive function score and cognitive decline,respectively.Stratified analysis was performed to explore whether the associations varied among different body mass index groups. Results Among the surveyed older adults,1 119(57.06%) were diagnosed as with cognitive decline by minimum mental state examination.The results of multivariate linear regression analysis showed that after adjusting for possible confounding factors in turn,the score of chili intake had a stable positive correlation with cognitive function score(β=0.086,95%CI=0.010-0.163,P=0.027).Compared with those who did not have chili intake,the following groups had increased cognitive function scores:those with low chili intake(β=0.559,95%CI=0.023-1.096,P=0.041) or moderate chili intake(β=0.641,95%CI=0.075-1.207,P=0.026),those who had chili intake 1-5 days per week(β=0.994,95%CI=0.430-1.558,P=0.001),those who began chili intake over the age of 18(β=0.797,95%CI=0.172-1.422,P=0.013),those who began chili intake at the age of 18 and under(β=0.476,95%CI=0.045-0.907,P=0.031),those had mild chili intake(β=0.471,95%CI=0.047-0.896,P=0.030),and those who had high chili intake(β=0.982,95%CI=0.275-1.689,P=0.007).Logistic regression analysis showed that chili intake 1-5 days per week(OR=0.634,95%CI=0.474-0.849,P=0.002) was associated with a lower risk of cognitive decline than those without chili intake.Stratified analysis showed that the positive association between chili intake and cognitive function was more significant in the group with body mass index ≥24 kg/m²,and the higher score of chili intake corresponded to the lower risk of cognitive decline(OR=0.924,95%CI=0.875-0.975,P=0.004). Conclusions Appropriate chili intake may have a protective effect on cognitive function in the elderly population,especially in the overweight and obese elderly.Prospective studies are needed to further verify the causal relationship.

Objective To investigate the expression of opioid growth factor(OGF),OGF receptor(OGFR),P16,and P21 in elderly patients with colorectal cancer and their correlations with clinicopathological characteristics. Methods Bioinformatic analysis of OGFR was performed via differential expression analysis,survival analysis,single-cell RNA sequencing,and spatial transcriptomics.A total of 167 elderly patients with colorectal cancer were enrolled,with 30 fresh tissue samples collected from January 2023 to December 2024 and 137 paraffin-embedded tissue samples collected from January 2016 to January 2020.All the specimens were collected from confirmed cases at the First Affiliated Hospital of Hebei North University.OGF expression in the 30 fresh tissue samples was detected by ELISA,while the protein levels of OGFR,P16,and P21 were measured by Western blot.Immunohistochemistry was employed to assess OGFR,P16,and P21 expression in the 137 paraffin-embedded samples.SPSS 20.0 was used to analyze the correlations of OGFR,P16,and P21 expression with clinicopathological characteristics.The Spearman method was adopted to evaluate correlations among OGFR,P16,and P21 expression.Overall 5-year survival and differences between groups were assessed by Kaplan-Meier survival curves and the log-rank test. Results Differential expression analysis revealed that the expression of OGFR was upregulated in colorectal cancer tissue samples(P<0.001).Survival analysis revealed a significant correlation between high OGFR expression and poor prognosis(disease-free interval:P=0.019,disease-specific survival:P=0.015).Single-cell and spatial omics analyses indicated that OGFR was primarily expressed in tumor cells.ELISA results showed that compared with that in the adjacent normal tissue,the OGF expression levels elevated in pathological stages Ⅰ(P<0.001),Ⅱ(P=0.042),Ⅲ(P<0.001),and Ⅳ(P=0.001).Western blot results demonstrated that compared with that in the adjacent normal tissue,the protein level of OGFR was up-regulated in pathological stages Ⅰ(P<0.001),Ⅱ(P=0.047),Ⅲ(P=0.045),and Ⅳ(P=0.012).Compared with those in the adjacent normal tissue,the protein levels of P16 and P21 were down-regulated in pathological stages Ⅰ(P=0.001,P<0.001),Ⅱ(P=0.031,P=0.008),Ⅲ(P=0.026,P=0.024),and Ⅳ(P=0.018,P=0.015).Immunohistochemistry results demonstrated that OGFR had higher expression in cancer tissue samples than in adjacent normal tissue samples(P<0.001),and its expression level was correlated with T stage(P=0.047),lymph node metastasis(P=0.035),TNM stage(P=0.013),differentiation degree(P=0.043),and distant metastasis(P=0.030).Conversely,the protein levels of P16 and P21 were lower in cancer tissue samples(all P<0.001).P16 expression was correlated with T stage(P=0.009),lymph node metastasis(P=0.035),TNM stage(P=0.001),and differentiation degree(P=0.021).P21 expression showed correlations with T stage(P=0.009),lymph node metastasis(P=0.002),and TNM stage(P=0.046).The 5-year overall survival rate of the 137 elderly patients with colorectal cancer was 55.47%.The patients with positive expression of OGFR had a lower 5-year overall survival rate than those with negative expression(P=0.001).The patients with negative expression of P16 or P21 had a lower 5-year overall survival rate than those with positive expression(P16:P<0.001;P21:P=0.007). Conclusions In the colorectal cancer tissue of elderly patients,the expression of OGFR was negatively correlated with that of P16 and P21.The OGF-OGFR signaling pathway may promote colorectal cancer progression by suppressing P16 and P21 expression.The combined detection of these markers shows potential as a valuable reference for assessing disease progression.

Objective To explore the clinical significance of roundabout guidance receptor 4(ROBO4) and protocadherin 12(PCDH12) in the human serum in the screening,diagnosis,and staging of colorectal cancer(CRC) through proteomics and bioinformatics,thus providing a novel screening method with high specificity,sensitivity,and accuracy for the early screening and diagnosis of CRC in clinical practice. Methods Proteomics and bioinformatics approaches were employed to select the target proteins ROBO4 and PCDH12,and the correlation between ROBO4 and PCDH12 was analyzed.A total of 121 CRC patients(cancer group),112 patients with polyps(polyp group),and 108 healthy volunteers(normal group) who received treatment or physical examination from January 2023 to June 2024 were enrolled in this study.ELISA was employed to determine the expression levels of ROBO4,PCDH12,carcinoembryonic antigen(CEA),and carbohydrate antigen 199(CA199) in the samples of each group.The expression levels of the four proteins in the serum were compared among the three groups of patients mentioned above,and the relationships of the serum levels of ROBO4 and PCDH12 with the pathological characteristics of the CRC patients were analyzed.Furthermore,the receiver operating characteristic(ROC) curves were established to evaluate the diagnostic efficacy of the serum levels of ROBO4 and PCDH12 for CRC patients in early and progressive stages. Results The target proteins ROBO4 and PCDH12 were screened out through proteomics.Bioinformatics analysis showed that ROBO4 and PCDH12 were highly expressed in CRC patients.Further correlation analysis revealed a positive correlation between ROBO4 and PCDH12 in CRC(R=0.870,P<0.001).The ELISA results of clinical samples showed that compared with the normal group and polyp group,the cancer group presented elevated expression levels of ROBO4 and PCDH12(all P<0.001).There was no statistically significant difference in the expression level of ROBO4 or PCDH12 between the normal group and the polyp group(P=0.586,P=0.550).The ROC curves showed that the diagnostic efficacy of ROBO4,PCDH12,and ROBO4+PCDH12 was 0.787,0.757,and 0.812,respectively,all of which were higher than the diagnostic efficacy of CEA and CA199.Further analysis of serum levels of ROBO4 and PCDH12 with the pathological data of CRC showed that the serum level of ROBO4 was correlated with differentiation degree(P=0.013),pathological stage(P=0.002),lymph node metastasis(P=0.001),and distant metastasis(P=0.026).The serum level of PCDH12 was correlated with differentiation degree(P=0.043),pathological stage(P=0.012),and lymph node metastasis(P=0.001).The ROC curves showed that the diagnostic efficacy of ROBO4,PCDH12,and ROBO4+PCDH12 was 0.637,0.758,and 0.787 for early CRC and 0.872,0.757,and 0.882 for progressive CRC,respectively. Conclusions The serum levels of ROBO4 and PCDH12 demonstrate high diagnostic efficacy for CRC patients and are correlated with the pathological features of CRC.The two proteins are expected to be novel serum biomarkers for the diagnosis and screening of CRC.

Objective To analyze the relationship between combined lifestyle and cognitive function among older adults in rural communities. Methods A multi-stage stratified whole-group sampling method was employed to select older adults aged ≥60 years in four villages of Chongming District,Shanghai as the participants.Their somatic indicators were measured by trained personnel,and data on demographic characteristics,cognitive function,and lifestyle were collected through face-to-face interviews.Binary Logistic regression and trend tests were adopted to explore the relationship between combined lifestyle and cognitive function.Additionally,the subgroup analysis and the test for multiplicative interactions were conducted for the relationship between combined lifestyle and cognitive function.Furthermore,sensitivity analysis was performed to verify the robustness of the results. Results A total of 1 449 older adults were investigated,and 1 408 were effective.The older adults showed the average age of(73.25±5.69) years and the cognitive impairment prevalence of 15.63%.Among them,163 older adults lived unhealthy lifestyles,accounting for 11.6%.After adjusting for demographic factors,disease history,and medication history,moderate lifestyles(OR=0.609,95%CI=0.394-0.944,P=0.026) and healthy lifestyles(OR=0.377,95%CI=0.212-0.672,P=0.001) had negative associations with the prevalence of cognitive impairment.Among combined lifestyle components,dietary diversity(OR=0.404,95%CI=0.211-0.771,P=0.006),physical activity intensity(OR=0.460,95%CI=0.235-0.901,P=0.024),and sleep duration(OR=0.611,95%CI=0.416-0.897,P=0.012) had negative associations with the prevalence of cognitive impairment.The trend test revealed that the negative association between cognitive impairment and combined lifestyle enhanced with the sequential increase in combined lifestyle score(P_{for trend}=0.011).The subgroup analysis revealed an interaction effect between gender and lifestyle subgroups,and a healthy lifestyle exerted a protective effect on women(P_{for interaction}=0.013). Conclusions Among older adults in rural communities,cognitive impairment has a significantly negative association with healthy lifestyles.As the combined lifestyle score increases,the risk of developing cognitive impairment shows a decreasing trend,which suggests that comprehensive lifestyle interventions may have a protective effect on cognitive function.

Objective To investigate the characteristics of symptom clusters and sentinel symptoms in lung cancer patients before and at the completion of radiotherapy,thus providing a theoretical basis for targeted symptom management. Methods A convenience sampling method was used to recruit patients from the Department of Radiotherapy at Yunnan Cancer Hospital between January and October 2024.A general information questionnaire and the Anderson symptom inventory-lung cancer module(revised) were employed for data collection.The exploratory factor analysis was performed for the symptoms with an incidence rate greater than 20% one day before and at the completion of radiotherapy to identify symptom clusters at different time points.The Apriori algorithm was then adopted to model the associations among symptoms within each cluster and to identify the sentinel symptoms. Results A total of 244 patients were included in this study.Three symptom clusters existed before radiotherapy:psychological symptom cluster(sadness,distress,and sleep disorder),lung cancer-related respiratory symptom cluster(cough,expectoration,chest tightness,and shortness of breath),and perception symptom cluster(pain and numbness).Distress and cough were the sentinel symptoms of psychological symptom cluster and lung cancer related respiratory symptom cluster,respectively.At the end of radiotherapy,the psychological symptom cluster and lung cancer-related respiratory symptom cluster remained unchanged.The radiotherapy side effect symptom cluster(dry mouth,pain,loss of appetite) appeared,and the perception symptom cluster expanded to encompass 5 items(amnesia,drowsiness,fatigue,nausea,and constipation).Pain and drowsiness were the sentinel symptoms of radiotherapy side effect symptom cluster and perception symptom cluster,respectively. Conclusions There are several symptom clusters in lung cancer patients before and at the completion of radiotherapy,and sentinel symptoms exist in symptom clusters at different periods.Nurses can take sentinel symptoms as the intervention targets to formulate effective and targeted nursing measures and reduce the incidence or severity of other symptoms in the cluster,thus diminishing the whole symptom cluster and reducing the burden of patients’ symptoms.

Objective To assess the human papillomavirus(HPV)testing and infection status among men who have sex with men(MSM)using geosocial networking apps in China and explore the influencing factors,thus providing evidence for designing effective prevention and control strategies for this population. Methods The participants were recruited via the Blued7.23 app from July 15 to August 2,2024.Univariate and multivariate Logistic regression models were used to analyze the factors associated with HPV testing and infection among MSM. Results Among the 1 080 participants,23.3% had undergone HPV testing,and 43.3% of those tested were positive for HPV.The main reason for HPV testing was the presence or suspicion of HPV-related symptoms(n=224,58.9%).The multivariate Logistic regression analysis showed that the MSM who had higher levels of HPV knowledge,had higher income,and self-identified as homosexuals were more likely to undergo HPV testing.Moreover,the MSM who had higher levels of HPV knowledge,self-identified as homosexuals,had sex with women in the past six months,and had infected human immunodeficiency virus were identified as high-risk groups for HPV infection. Conclusions In China,the HPV testing rate is low among MSM using geosocial networking apps.However,the prevalence of HPV infection remains high.Therefore,the government and communities should enhance HPV knowledge dissemination,promote HPV testing,and improve the accessibility and quality of HPV testing services.These measures can increase the testing coverage and effectively control the spread of HPV among MSM.