Objective To study the five-year survival status and influencing factors of elderly patients with lung cancer complicated with chronic obstructive pulmonary disease (COPD). Methods A cohort study was conducted to follow up 450 patients with lung cancer and chronic obstructive pulmonary disease who were hospitalized in our hospital from January 2018 to December 2023. The endpoint of the follow-up was the end of a five-year period or death. The Life Tables method was used to calculate survival rates and plot survival curves. The Cox proportional hazards model was used to analyze the influencing factors of five-year survival. Results The results indicated that the overall five-year survival rate of patients was 4.89%, and it decreased year by year. Cox regression analysis showed that age, gender, family functioning, and psychological status significantly influenced patient survival rate (all P<0.05). Stratified analysis found that the smoking status, family functioning, and psychological status of male patients all had an impact on survival rate (all P<0.05), while the psychological status of female patients had a more significant impact on survival (P=0.008). Conclusion This study provides a scientific basis for comprehensive intervention of elderly lung cancer patients with COPD. It is recommended that clinical attention should be paid to psychological and family factors to improve patient prognosis.

ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

Objective:To study and evaluate the efficacy of hemolytic protection method for extracting DNA from Plasmodium dried blood spots (DBS). Methods:Centrifugal column method, resin method, and hemolytic protection method were used, respectively, to extract DNA from the same batch of DBS (1.0, 10.0, 100.0, 1 000.0, 10 000.0 parasites/μl blood) that were prepared and preserved according to unified standards. Dilute DNA extracted from 1.0 parasites/μl blood DBS by 10 times to prepare 0.1 parasites/μl blood template DNA. Nested PCR was used to detect templates prepared by the three different methods (with template DNA of ≥1.0 parasites/μl blood DBS repeated 3 times). The limit of detection (LOD) and detection rate were compared by repeating the detection of 0.1 parasites/μl blood template DNA for 30 times. The ultra-micro ultraviolet spectrophotometer was used to measure the concentration and purity of DNA extracted from DBS (10.0, 100.0, and 1 000.0 parasites/μl blood), three times each, and the DNA recovery amount was calculated. Meanwhile, the applicability and cost of different methods were analyzed.Results:The nested PCR detection rate of the template DNA of ≥1.0 parasite/μl blood DBS extracted by the three methods was all 3/3. For the 0.1 parasite/μl blood template DNA, the nested PCR detection rates were 0 (0/30), 73.33% (22/30), and 100% (30/30), respectively, and there was a significant difference (χ 2 = 65.95, P < 0.001). The LOD was 1.0, 0.1, 0.1 parasite/μl blood, respectively. When comparing the DNA concentration, purity, and recovery amount extracted by the 3 methods, all showed significant differences ( H = 23.25, 17.50, 23.25, P < 0.001). The centrifugal column method has 9 steps, an extraction time of 2.5 h, and the cost for each sample was 33.32 yuan. The resin method has 7 steps, an extraction time of 20.0 h, and the cost for each sample was 7.11 yuan. The hemolytic protection method has 4 steps, an extraction time of 1.0 h, and the cost for each sample was 1.96 yuan. Conclusion:Hemolytic protection method is an efficient, sensitive, rapid, and economical method for extracting DNA from Plasmodium DBS.

Objective To investigate the effects of integrating pelvic floor magnetic stimulation and medium-frequency pulsed electrotherapy with biofeedback on alleviating obstructed defecation symptoms in patients diagnosed with pelvic floor spasm syndrome.Methods A total of 133 patients diagnosed with pelvic floor spasm syndrome at the Gastrointestinal Surgery Outpatient and Inpatient Departments of the Third Hospital of Hebei Medical University between January 2017 and January 2025 were enrolled in this study.All patients underwent two sessions of conventional biofeedback therapy.According to the additional treatments they received,the patients were catego-rized into four groups:the Control group(received biofeedback therapy only);the Magnetic Stimulation group(received two additional sessions of pelvic floor magnetic stimulation);the Electrical Stimulation group(received two additional sessions of medium-frequency pulse electrical therapy);and the Magnetic-Electric Combined group(received two additional sessions of both pelvic floor magnetic stimulation and medium-frequency pulse electrical therapy).The improvement in obstructed defecation symptoms was evaluated before treatment,after treatment,and at the 3-month follow-up across all four groups.Results A total of 133 patients diagnosed with pelvic floor spasm syndrome were enrolled in this study.After a 3-month follow-up period,8 patients were lost to follow-up(6.0%).Consequently,125 patients completed the follow-up and were included in the final analysis.These patients were distributed across four groups:the control group(n=32),the magnetic stimulation group(n=30),the electrical stimulation group(n=31),and the combined magneto-electric stimulation group(n=32).Compared with biofeedback therapy alone,combination therapy led to a greater reduction in the need for manual assistance during defecation,the sensation of incomplete evacuation,and the requirement for straining during defecation,with the most pronounced improvements observed in the combined magneto-electric stimulation group.Conclusions The integration of biofeedback,pelvic floor magnetic stimulation,and medium-frequency pulse electrotherapy can significantly alleviate symptoms including straining during defecation,the sensation of incomplete bowel evacua-tion,and reduce the need for manual assistance.Moreover,this combined approach contributes to the stabilization and maintenance of therapeutic effects in the short term.

Objective To develop a rapid and accurate ultra-performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS)method for the detection of etomidate,metomidate,propoxate,and isopropoxate in human hair.Methods Hair samples containing etomidate,metomidate,propoxate,and isopropoxate were extracted with methanol containing the internal standard orthoxine,filtered with a 0.22 μm organic filter membrane and detected vio UPLC-MS/MS.All components were separated by using a gradient elution procedure consisting of 0.01％formic acid(1 mmol/L ammonium acetate)and acetonitrile.Positive electrospray ionization was performed using multiple reaction monitoring(MRM)mode.Results The linear relationships of etomidate,metomidate,propoxate,and isopropoxate were good in the range of 0.01～1 ng/mg(r ≥ 0.997 9),with recovery rates ranging from 87.9％to 101.5％.The accuracy was between 80.0％and 110.0％.The intra-day and inter-day relative standard deviations(RSD)were 2.9％～9.6％and 3.6％～19.9％.Conclusion This method is easy to operate and has high recovery efficiencies.It is sufficiently simple and sensitive to be applied to detect etomidate,metomidate,propoxate,and isopropoxate in hair.

Objective To compare the consistency of voice parameters collected by commonly used smart-phone brands in China and professional recording equipment,and to study whether smartphones can be used for voice research.Methods A total of 67 normal subjects were selected for voice recording using six different smart-phone brands(via the"Active Health"screening APP from the National Key Research and Development Program)and professional recording equipment.Acoustic voice parameters such as fundamental frequency parameters,fre-quency variation parameters,amplitude variation parameters,formant parameters,and energy parameters were ex-tracted from the vowels/a/,/i/,and/u/.A one-way ANOVA test and Tukey's HSD post-hoc comparisons were conducted on the independent variables.Results There were no significant differences between smartphones and professional recording equipment in terms of fundamental frequency parameters such as median F0,mean F0,max F0 and min F0;frequency parameters such as jitter local,jitter local absolute,jitter rap,jitter ppq5,and jitter ddp;amplitude parameters such as shimmer local,shimmer local dB,shimmer apq3,shimmer apq5,shimmer apq11,and shimmer dda;and formant parameters such as F1,F2,F3,and F4.However,significant differences were found in energy parameters such as mean energy(F=31.171,P＜0.001),max energy(F=34.193,P＜0.001),and min energy(F=5.453,P＜0.001)between smartphones and professional recording equipment.Conclusion The smartphones using the"Active Health"screening app from the National Key Research and Development Program can replace professional recording equipment for voice research.However,caution should be exercised when selec-ting energy-related acoustic parameters.

The aim of this study is to identify the cause of death of a South China tiger cub at the Meihuashan breeding institute of Fujian Province.Pathogenic bacteria were isolated and cultured from liver,spleen,lung and other tissue samples of the dead South China tiger aseptically.The iso-lated bacteria were identified through morphological observation,biochemical characterization,sequence analysis of housekeeping gene gyrB,virulence gene detection,animal pathogenicity test and drug sensitivity test.A pathogenic Aeromonas hydrophila strain,designated FJ/Tiger-201809 was successfully isolated from the trachea of dead South China tiger.The nucleotide sequence ho-mology between the isolate and 11 strains of Aeromonas gyrB ranged from 91.2％to 99.1％,with the highest homology of 99.1％observed with Aeromonas hydrophila(AF208251.1).Genetic evo-lution analysis showed that the isolated strain FJ/Tiger-201809 was in the same evolutionary branch as other reference strains of Aeromonas hydrophila and was closely related.The pathoge-nicity test in mice showed artificial infection of mice with the strain resulted in varying degrees of lesions in several organs of the mice,and the median lethal dose(LD50)was 1 × 107.8 CFU/mL.Virulence gene test results showed that the isolate FJ/Tiger-201809 carried two virulence genes,aer and act.The results of drug sensitivity test showed that FJ/Tiger-201809 was highly sensitive to enrofloxacin and ampicillin among 18 commonly used antibiotics,relatively sensitive to penicil-lin G and doxycycline,and resistant to the other 14 antibiotics.In conclusion,this study isolated and identified a strain of Aeromonas hydrophila from a dead South China tiger with multiple drug resistance and strong pathogenicity,which provided an important reference for the prevention and control of bacterial diseases in South China tiger.

Objective:To explore the association between healthy lifestyle and co-morbidity of overweight/obesity and myopia in adolescents.Methods:Based on the Shanghai Municipal Dynamic Cohort of Student Common Diseases, we used the group-based trajectory model (GBTM) to establish a model for the development of adolescents' healthy lifestyle trajectories aged 11-14 years and the Cox proportional hazard regression model to assess the effects of different healthy lifestyles on the co-morbidity of overweight/obesity and myopia in adolescents.Results:A total of 2 331 adolescents were included in the analysis. The average follow-up time was 1.87 years, with age of (11.44±0.45) years at the initial follow-up, among 1 106 boys and 1 225 girls. During the 2-year follow-up period, the incidence rates of overweight/obesity, myopia, and co-occurrence of overweight/obesity and myopia were 25.9%, 77.0%, and 13.7%, respectively. GBTM identified 3 healthy lifestyles: the unhealthy group (7.3%), the relatively healthy group (50.2%), and the healthy group (42.5%). The risk of overweight/obesity was lower in the relatively healthy group than in the unhealthy group ( HR=0.694, 95% CI: 0.508-0.947). There was no statistically significant association between the GBTM lifestyle subgroups and myopia ( P>0.05). The risk of co-occurrence of overweight/obesity and myopia was lower in both relatively healthy group and healthy group than in the unhealthy group (relatively healthy group: HR=0.515, 95% CI: 0.348-0.763; healthy group: HR=0.571, 95% CI: 0.384-0.849). Sensitivity analyses showed this result to be stable for myopia, overweight/obesity and myopia co-morbidities. Conclusion:Persistent unhealthy lifestyle increases the risk of overweight/obesity and myopia co-occurrence in adolescents.

Objective To investigate the correlation between Traditional Chinese Medicine(TCM)constitution types and disease progression in elderly patients with hypertension complicated by heart failure(HF).Methods A total of 200 eligible elderly patients treated in our hospital from June 2021 to June 2023 were recruited,and based on TCM constitution types,they were classified as balanced constitution(n=20,10.0％),qi-deficiency(n=35,17.5％),yang-deficiency(n=30,15.0％),yin-deficiency(n=21,10.5％),phlegm-dampness(n=35,17.5％),damp-heat(n=16,8.0％),blood-stasis(n=25,12.5％),qi-stagnation(n=11,5.5％),and inherited special constitu-tion(n=7,3.5％).TCM constitution,blood pressure,and cardiac function indicators were as-sessed.All patients were followed up for 1 year,and acute exacerbations was recorded to analyze disease progression.Blood pressure,cardiac function indicators,and acute exacerbation frequency were compared among different TCM constitution types,and Spearman correlation analysis was used to assess the relationship between TCM constitution types and the frequency of acute exacer-bations.Results Significant differences were observed in SBP,DBP and frequency of acute exacer-bations among elderly hypertension patients with HF having different TCM constitution types(P＜0.01).The patients with a phlegm-dampness constitution had obviously higher SBP,DBP,mean arterial pressure,stroke volume,central venous pressure and frequency of acute exacerba-tions than those with other constitution types,while the patients with a balanced constitution had the notably highest left ventricular ejection fraction(P＜0.01).Spearman correlation analysis showed that balanced,phlegm-dampness,and blood-stasis constitutions were positively correlated with the frequency of acute exacerbations in these patients(r=0.480,r=0.375,r=0.295,P＜0.01),whereas qi-deficiency constitution was negatively correlated(r=-0.235,P＜0.01).Conclusion Elderly patients with hypertension and HF are predominantly of the qi-deficiency,phlegm-dampness,and yang-deficiency constitution types.Among them,the patients with phlegm-dampness constitution exhibit notably higher blood pressure and cardiac function indicators.TCM constitution types such as balanced and phlegm-dampness are closely related to disease progres-sion in these patients.

Objective:To construct a questionnaire to measure parents′ behavioral and social drivers (BeSD) in administering rotavirus vaccine to their children, and to test the reliability and validity of the questionnaire.Methods:A questionnaire was constructed to investigate the BeSD of rotavirus vaccination among parents based on the WHO BeSD questionnaire. A survey was conducted among parents of children aged 0-3 years old in vaccination clinics from 15 community health service centers in Nanjing City of Jiangsu Province, Yuhuan and Kaihua Counties of Zhejiang Province, Anqing City of Anhui Province, and Yangpu District of Shanghai City from April to May in 2024. Exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) were used to analyze the reliability (internal consistency reliability, combination reliability and split half reliability) and validity (structural validity, convergence validity and discriminative validity) of the questionnaire.Results:A total of 850 questionnaires were collected, of which 614 were valid, with an effective response rate of 72.20%. The EFA resulted in the refinement of the rotavirus vaccine BeSD questionnaire from 37 to 35 items (including 24 core items), and suggested a five-factor structure including the perceptions of vaccine, confidence in vaccine, social process of vaccination, motivation for vaccination, and practical problems of vaccination. The cumulative variance contribution rate reached 56.27%. The CFA confirmed an ideal five-factor model fit (GFI=0.82,CFI=0.86,PNFI=0.67,RMSEA=0.07). The AVE of each dimension was greater than 0.50. The AVE square root of each dimension of the questionnaire was greater than its correlation coefficient. The Cronbach′s α coefficient of the total questionnaire was 0.79.Conclusion:The developed BeSD questionnaire of rotavirus vaccine has good reliability and validity, and can be used to measure parents′ behavioral and social factors in administering rotavirus vaccine to their children.