Abstract To improve adolescent health behavior, the study summarizes and analyzes the performance, pathways of transmission, and influencing factors of the intergenerational transmission of adolescent health behaviors from the perspective of intergenerational transmission. The study emphasizes the need to deepen research on the intergenerational transmission of adolescent health behaviors, promote multidisciplinary and cross team collaboration, and shift adolescent health care from individual focused care to a holistic approach that prioritizes family and community culture. Simultaneously, an action framework should be established to block the intergenerational transmission of health risk behaviors, with a focus on childhood and adolescence. Additionally, parent-child participatory health education and health promotion activities should be carried out under a tripartite coordinated intervention model involving the community, school, and family, collectively fostering the development of healthy behaviors among adolescents.

ObjectiveTo investigate the association of alpha-fetoprotein （AFP） and prealbumin （PAB） with the 90-day prognosis of patients with hepatitis B virus-related acute-on-chronic liver failure （HBV-ACLF）， as well as the difference in 90-day prognosis between the patients with different levels of AFP and PAB. MethodsA total of 371 HBV-ACLF patients who were hospitalized in The General Hospital of Western Theater Command from January 2018 to January 2023 were enrolled， and according to the follow-up results on day 90 after discharge， they were divided into survival group with 216 patients and death group with 155 patients. The medical record system was used to collect general data， AFP， PAB， and other related laboratory markers. The t-test was used for comparison of normally distributed continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for comparison between two groups. The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups， and the Kruskal-Wallis H test was used for comparison between multiple groups and further comparison between two groups. The chi-square test was used for comparison of categorical data between groups. The multivariate logistic regression analysis was used to identify the influencing factors for the prognosis of HBV-ACLF patients. The receiver operating characteristic （ROC） curve was plotted for AFP and PAB to determine their cut-off values. The Kaplan-Meier method was used to plot survival curves， and the Log-rank test was used for comparison. ResultsCompared with the death group， the survival group had significantly higher levels of hemoglobin （Hb）， PAB， AFP， and platelet count （PLT） （all P<0.05） and significantly lower age， total bilirubin （TBil）， white blood cell count （WBC）， cystatin， creatinine， urea， international normalized ratio （INR）， Model for End-Stage Liver Disease （MELD） score， proportion of patients with Child-Pugh class C， and incidence rates of ascites and hepatic encephalopathy （all P<0.05）. The multivariate logistic regression analysis showed that PAB （odds ratio ［OR］=0.985， 95% confidence interval ［CI］： 0.972‍ ‍—‍ ‍0.998， P=0.024）， AFP （OR=0.998， 95%CI： 0.996‍ ‍—‍ ‍1.000， P=0.028）， PLT （OR=0.989， 95%CI： 0.982‍ ‍—‍ ‍0.996， P=0.003）， age （OR=1.046， 95%CI： 1.018‍ ‍—‍ ‍1.075， P=0.001）， TBil （OR=1.004， 95%CI： 1.002‍ ‍—‍ ‍1.006， P<0.001）， and WBC （OR=1.237， 95%CI： 1.110‍ ‍—‍ ‍1.379， P<0.001） were independent influencing factors for 90-day prognosis in HBV-ACLF patients. According to the cut-off values of AFP and PAB on ROC curves， the patients were divided into group A with 102 patients （AFP≥73.19 ng/mL and PAB≥22.55 mg/L）， group B with 170 patients （AFP≥73.19 ng/mL and PAB<22.55 mg/L； AFP<73.19 ng/mL and PAB≥22.55 mg/L）， and group C with 99 patients （AFP<73.19 ng/mL and PAB<22.55 mg/L）. There were significant differences between these three groups in age， Hb， INR， MELD score， and Child-Pugh class （all P<0.05）. The Kaplan-Meier survival analysis showed that group A had a significantly higher 90-day cumulative survival rate than groups B and C （χ²=19.825， P<0.001）. ConclusionAFP combined with PAB can better predict the 90-day prognosis of HBV-ACLF patients， and patients with high levels of AFP and PAB tend to have a lower mortality rate on day 90.

Objective:To explore the effect of long noncoding RNA ANK3DT (lncRNA ANK3DT) on DNA damage repair ability and radiosensitivity of cervical cancer HeLa cells.Methods:The effect of lncRNA ANK3DT on the repair efficiency of DNA double-strand breaks (DSBs) was analyzed by the pre-constructed quantitative assay system based on the CRISPR/Cas9 system targeting homologous recombination (HR) and non-homologous end-joining (NHEJ) repair. Clone formation assay, flow cytometry, and immunofluorescence were used to detect the effects of down-regulation of lncRNA ANK3DT on cellular radiosensitivity, apoptosis and G 2/M phase arrest, and DSB repair after X-ray irradiation. Western blot, qPCR, and dual fluorokinase reporter gene plasmid was used to detect the effects of lncRNA ANK3DT on the HR repair-related protein CtIP expression and transcriptional regulation. Results:Down-regulation of lncRNA ANK3DT significantly inhibited HR repair and had no effect on NHEJ repair. The expression of lncRNA ANK3DT was significantly increased in HeLa cells at 24, 48, 72 h after X-ray irradiation ( t=-23.39, -88.83, -52.42, P<0.05). Down-regulation of lncRNA ANK3DT inhibited X-ray-induced DSB repair and increased apoptosis levels ( t=-14.63, P <0.05), prolonged G 2/M phase block ( t=-19.50, P <0.05), increased the radiosensitivity of HeLa cells (radiosensitization ratio=1.21), inhibited the CtIP promoter activity and decreased its mRNA and protein expression. Conclusions:lncRNA ANK3DT affects HR repair by regulating CtIP transcription, and down-regulation of lncRNA ANK3DT increases the radiosensitivity of HeLa cells by inhibiting DSB repair after ionizing radiation.

Objective:To utilize single-nucleus RNA sequencing(snRNA-seq) technology to investigate the primary cell subpopulations in human limb venous malformations (VMs) tissue and the role of the key gene RhoJ.Methods:Surgical resection specimens of VMs tissues and surrounding normal vein tissues were collected from 100 clinically diagnosed and screened patients with limb VMs at the Department of Hemangioma Surgery of the Third Affiliated Hospital of Zhengzhou University from January 2021 to December 2023. (1) Transcriptome analysis: Three patient samples were randomly selected for snRNA-seq studies, with the surgically removed VMs tissue serving as the experimental group and the surrounding normal vein tissue as the control group. A gene expression matrix for cell nuclei was established, followed by data quality control, dimensionality reduction, clustering, and cell type annotation. Cell-to-cell communication analysis was performed using the R language CellChat package to identify dominant cell subpopulations. The FindMarkers function was utilized to screen for differentially expressed genes (DEGs) between the dominant cell subpopulations of the experimental and control groups, and functional enrichment analysis was conducted. (2) Tissue experiments: An additional 35 patient samples from both the experimental and control groups were randomly selected. The mRNA and protein expression levels of the RhoJ gene were measured using real-time quantitative PCR (RT-qPCR) and Western blotting, respectively. (3) Validation experiments with human umbilical vein endothelial cells (HUVECs): HUVECs were transfected with pcDNA3.1-NC (blank control) and pcDNA3.1-RhoJ (plasmid expression vector carrying the RhoJ gene), respectively. The biological behavior differences between the two groups of cells were examined using the CCK-8 cell proliferation assay, Transwell invasion assay, and Matrigel angiogenesis assay. Measurement data conforming to a normal distribution were expressed as Mean±SD, and comparisons between the two groups were performed using an independent samples t-test. Results:Through CellChat intercellular communication analysis, it was discovered that endothelial cells were the predominant cell subpopulation in both the experimental and control groups, exhibiting strong communication links with other cell subpopulations. In the analysis of DEGs, it was found that the RhoJ gene in endothelial cells was significantly involved in the biological processes of angiogenesis and regulation. In tissue experiments, RT-qPCR and Western bloting results indicated that the relative expression levels of RhoJ mRNA (4.48±1.29 vs. 1.01±0.17) and protein (1.22±0.03 vs. 0.51±0.20) in the experimental group were significantly higher than those in the control group, with statistically significant differences ( P<0.01 for both). The results of the HUVECs validation experiment showed that the cell proliferation, invasion, and angiogenesis abilities of the pcDNA3.1-RhoJ group were significantly enhanced compared to the pcDNA3.1-NC group. Conclusion:Endothelial cells represent the dominant cell subpopulation during the occurrence and locally invasive progression of VMs, playing a crucial role in this process. The RhoJ gene is significant in regulating the biological behavior of VMs endothelial cells.

Objective To investigate the association of alpha-fetoprotein(AFP)and prealbumin(PAB)with the 90-day prognosis of patients with hepatitis B virus-related acute-on-chronic liver failure(HBV-ACLF),as well as the difference in 90-day prognosis between the patients with different levels of AFP and PAB.Methods A total of 371 HBV-ACLF patients who were hospitalized in The General Hospital of Western Theater Command from January 2018 to January 2023 were enrolled,and according to the follow-up results on day 90 after discharge,they were divided into survival group with 216 patients and death group with 155 patients.The medical record system was used to collect general data,AFP,PAB,and other related laboratory markers.The t-test was used for comparison of normally distributed continuous data between two groups;a one-way analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for comparison between two groups.The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups,and the Kruskal-Wallis H test was used for comparison between multiple groups and further comparison between two groups.The chi-square test was used for comparison of categorical data between groups.The multivariate logistic regression analysis was used to identify the influencing factors for the prognosis of HBV-ACLF patients.The receiver operating characteristic(ROC)curve was plotted for AFP and PAB to determine their cut-off values.The Kaplan-Meier method was used to plot survival curves,and the Log-rank test was used for comparison.Results Compared with the death group,the survival group had significantly higher levels of hemoglobin(Hb),PAB,AFP,and platelet count(PLT)(all P<0.05)and significantly lower age,total bilirubin(TBil),white blood cell count(WBC),cystatin,creatinine,urea,international normalized ratio(INR),Model for End-Stage Liver Disease(MELD)score,proportion of patients with Child-Pugh class C,and incidence rates of ascites and hepatic encephalopathy(all P<0.05).The multivariate logistic regression analysis showed that PAB(odds ratio[OR]=0.985,95%confidence interval[CI]:0.972-0.998,P=0.024),AFP(OR=0.998,95%CI:0.996-1.000,P=0.028),PLT(OR=0.989,95%CI:0.982-0.996,P=0.003),age(OR=1.046,95%CI:1.018-1.075,P=0.001),TBil(OR=1.004,95%CI:1.002-1.006,P<0.001),and WBC(OR=1.237,95%CI:1.110-1.379,P<0.001)were independent influencing factors for 90-day prognosis in HBV-ACLF patients.According to the cut-off values of AFP and PAB on ROC curves,the patients were divided into group A with 102 patients(AFP≥73.19 ng/mL and PAB≥22.55 mg/L),group B with 170 patients(AFP≥73.19 ng/mL and PAB<22.55 mg/L;AFP<73.19 ng/mL and PAB≥22.55 mg/L),and group C with 99 patients(AFP<73.19 ng/mL and PAB<22.55 mg/L).There were significant differences between these three groups in age,Hb,INR,MELD score,and Child-Pugh class(all P<0.05).The Kaplan-Meier survival analysis showed that group A had a significantly higher 90-day cumulative survival rate than groups B and C(χ2=19.825,P<0.001).Conclusion AFP combined with PAB can better predict the 90-day prognosis of HBV-ACLF patients,and patients with high levels of AFP and PAB tend to have a lower mortality rate on day 90.

OBJECTIVE: To develop an artificial intelligence-assisted system for the automatic detection of the features of common 21 auricular points based on the YOLOv8 neural network. METHODS: A total of 660 human auricular images from three research centers were collected from June 2019 to February 2024. The rectangle boxes and features of images were annotated using the LabelMe5.3.1 tool and converted them into a format compatible with the YOLO model. Using these data, transfer learning and fine-tuning training were conducted on different scales of pretrained YOLO neural network models. The model's performance was evaluated on validation and test sets, including the mean average precision (mAP) at various thresholds, recall rate (recall), frames per second (FPS) and confusion matrices. Finally, the model was deployed on a local computer, and the real-time detection of human auricular images was conducted using a camera. RESULTS: Five different versions of the YOLOv8 key-point detection model were developed, including YOLOv8n, YOLOv8s, YOLOv8m, YOLOv8l, and YOLOv8x. On the validation set, YOLOv8n showed the best performance in terms of speed (225.736 frames per second) and precision (0.998). On the external test set, YOLOv8n achieved the accuracy of 0.991, the sensitivity of 1.0, and the F1 score of 0.995. The localization performance of auricular point features showed the average accuracy of 0.990, the precision of 0.995, and the recall of 0.997 under 50% intersection ration (mAP⁵⁰). CONCLUSION The key-point detection model of 21 common auricular points based on YOLOv8n exhibits the excellent predictive performance, which is capable of rapidly and automatically locating and classifying auricular points.
Humans ; Neural Networks, Computer ; Artificial Intelligence ; Acupuncture Points

Objective:This study aimed to identify key genes in endothelial cell (EC) associated with the pathogenesis and progression of human venous malformations (VMs) through bioinformatics analysis, providing potential biomarkers for early screening and targeted therapy of VMs.Methods:A case-control study was conducted using surgically resected tissue specimens from VMs patients at the Third Affiliated Hospital of Zhengzhou University (from September 2021 to September 2023), with malformed venous tissues as the experimental group and distal normal venous tissues as controls. Single-nucleus RNA sequencing (snRNA-seq) was performed on paired experimental and control samples from four VM patients. High-dimensional weighted gene co-expression network analysis (hdWGCNA), combined with gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG), and protein-protein interaction (PPI) network analysis, identified critical genes. Validation experiments included 15 additional VM cases and controls using reverse transcription quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry (IHC), and Western blot.Results:A total of 55 430 nuclei were captured using snRNA-seq, with 30 391 nuclei from the experimental group and 25 039 nuclei from the control group. Cluster analysis identified 22 distinct cell populations, which were annotated into 8 cell types. hdWGCNA revealed four modules associated with invasion, which were enriched in angiogenesis, integrin signaling, and cell adhesion according to GO analysis. KEGG pathway analysis indicated that the PI3K-AKT signaling pathway and focal adhesion are key regulatory mechanisms. PPI network analysis combined with cytoscape identified EGFL7, TEK, and FLT1 as key genes. RT-qPCR results demonstrated that the relative mRNA expression levels of these three genes in the experimental group (6.66±2.31, 1.86±0.62, 3.49±0.58) were significantly higher than those in the control group (1.05±0.14, 1.00±0.14, 1.06±0.25), with statistically significant differences ( t=9.37, 4.27, 11.20, P<0.05). Immunohistochemical analysis showed that the relative protein expression levels of these three genes in the cytoplasm of the experimental group (0.84±0.15, 0.68±0.14, 0.85±0.12) were also significantly higher than those in the control group (0.19±0.05, 0.23±0.06, 0.30±0.05), with statistically significant differences ( t=16.62, 5.93, 11.68, P<0.05). Western blot analysis confirmed that the relative protein expression levels of these three genes in the experimental group (0.35±0.04, 0.36±0.09, 0.31±0.04) were significantly higher than those in the control group (0.19±0.01, 0.13±0.02, 0.14±0.04), with statistically significant differences ( t=7.05, 4.61, 5.93, P<0.05). Conclusion:EGFL7, FLT1, and TEK in EC may play crucial roles in the occurrence and invasion of VMs.

To evaluate the clinical application value of magnetic bead sorting technology (MBS) using the T-cell select kit combined with an automatic cell sorter for isolating peripheral blood mononuclear cells (PBMCs) to realize automated specimen pre-processing and process optimization in T-SPOT.TB assay. A cross-sectional study was conducted involving 300 patients recruited from the first affiliated hospital of Air Force Medical University from March 2023 to July 2024. Among them, there were 167 males and 133 females. The age range of the patients was 18 to 65 years, with a median age of 49.0 (33.3, 59.0) years and a mean age of (46.4±13.9) years. 4 ml of anticoagulated whole blood samples in duplicate were collected from each patient. One fresh sample (0-4 h) was processed immediately using density gradient centrifugation (DGC) for PBMC isolation, while the other was processed using MBS method at either 0-4 h or 34-54 h post-collection. The cycles for cell enrichment step of the automatic cell sorter were adjusted from the conventional 4 cycles to 2 cycles. Statistical analysis was performed using Cohen′s Kappa test to evaluate the concordance of T-SPOT.TB results across all processing groups. The results showed that for fresh samples (0-4 h), the T-SPOT.TB results for samples processed with 4-cycle and 2-cycle MBS enrichment steps demonstrated positive concordance rates of 91.7% and 93.1%, negative concordance rates of 100% for both, overall concordance rates of 98.0% and 97.9%, and Kappa values of 0.94 and 0.95, respectively, compared with the reference results for paired samples processed using DGC. Correspondingly, the results for samples processed with the 4-cycle and 2-cycle MBS methods at 34-54 h post-collection yielded positive concordance rates of 90.0% and 81.3%, negative concordance rates of 95.1% and 100%, overall concordance rates of 93.0% and 82.9%, and Kappa values of 0.86 and 0.43, respectively, compared with the reference results for fresh samples processed using DGC. In conclusion,for 0-4 h samples, the T-SPOT.TB results from both 4-cycle and 2-cycle MBS enrichment protocols were highly consistent with the reference results from conventional DGC methods. However, for 34-54 h stored samples, results from the 4-cycle MBS method rather than the 2-cycle protocol exhibited significantly superior concordance with the reference results. The MBS method using T-Cell Select kit achieves automated sample pre-processing for T-SPOT.TB assay, reduces hands-on time, and provides a viable alternative with reliable results for long-stored specimens.

Objective To explore the barriers of implementing collaborative chronic wound management within medical consortium and provide references for ensuring continuous care for chronic wound patients.Methods Using purposive and snowball sampling methods,14 specialist nurses participating in chronic wound collaborative management across medical consortium in 7 provinces/municipalities(Beijing,Shanghai,Zhejiang,Sichuan,etc.)were selected for semi-structured interviews between December 2023 and January 2024.Data were analyzed via content analysis.Results Totally 4 themes and 8 subthemes were identified,including management and organizational level(the management system needs improvement;the function of medical information mutual recognition has not been fully realized),resource and technology level(there are insufficient nursing resources at the grassroots hospitals;the specialized nursing capabilities of wound nurses at grassroots hospitals need to be enhanced),patient and demand level(patients' trust is insufficient;patients' concepts are rigid),information technology and support level(the informatization construction of grassroots hospitals lags behind;the intelligence of information systems is insufficient).Conclusion Wound specialist nurses encounter various difficulties when participating in the coordinated management of chronic wounds within the medical consortium.The management and organizational system constraints hinder the effectiveness of the nursing coordination;the resource and technology gap affects the implementation of graded wound care in the region;the lack of patient trust and misconceptions intensify the resistance to coordination;the lagging in information technology construction limits the innovative development of nursing coordination.

Objective:To analyze the effects of fertilization methods, quality of inner cell mass (ICM) and trophectoderm (TE) on the sex ratio at birth (SRB) during single blastocyst transfer cycles.Methods:5 367 single blastocyst transfer cycles in in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET) of patients with successful pregnancy and delivery in Reproductive Medical Center of Zhanjiang Jiuhe Hospital from January 2010 to December 2022 were included in this retrospective cohort study. According to different types of embryos, the cycles were divided into fresh embryo group ( n=1 487, total population; n=1 258, ≤35 years old) and frozen-thawed embryo group ( n=3 880, total population; n=3 199, ≤35 years old). According to different fertilization methods, the cycles were divided into IVF group ( n=4 424, total population; n=3 667, ≤35 years old) and ICSI group ( n=943, total population; n=780, ≤35 years old). Chi-square test was used to analyze the differences of SRB among all the patients and the patients aged ≤35 years who underwent different embryo types and fertilization methods, and logistic regression was used to analyze the effects of ICM and TE ratings on SRB in each group of blastocysts. Results:There were 917 male infants (61.7%, 917/1 487) in the fresh embryo transfer group and 2 317 male infants (59.7%, 2 317/3 880) in frozen-thawed embryo transfer group. The SRB of the fresh embryo transfer group (160.9∶100) was slightly higher than that in frozen-thawed embryo transfer group (148.2∶100), but the difference was not statistically significant ( P=0.201). There were 2 732 male infants (61.8%, 2 732/4 424) in IVF group and 502 male infants (53.2%, 502/943) in ICSI group. The SRB of IVF group (161.5∶100) was significantly higher than that of ICSI group (113.8∶100), and the difference was statistically significant ( P<0.001). In female patients aged ≤35 years, the effects of different embryo transfer methods and different fertilization methods on SRB were consistent with the results of total population. Univariate logistic regression analysis showed that the SRB of patients with ICSI was lower than that of patients with IVF, except for 3-6AA. There were significant differences in SRB between ICSI cycles patients (88.8∶100; 72.8∶100; 156.1∶100) and IVF cycles patients (130.7∶100; 124.8∶100; 206.3∶100) when the blastocyst grade was 3-6BB ( OR=0.679, 95% CI: 0.516-0.896, P=0.006), 3-6AB( OR=0.583, 95% CI: 0.421-0.809, P=0.001) and 3-6BA OR=0.757, 95% CI: 0.585-0.979, P=0.034). Taking 3-6BB blastocysts as reference, the SRB of 3-6AA and 3-6BA blastocysts were higher (IVF: OR=1.527, 95% CI: 1.258-1.854, P<0.001; OR=1.579,95% CI: 1.341-1.859, P<0.001; ICSI: OR=2.566, 95% CI: 1.661-3.966, P<0.001; OR=1.758, 95% CI: 1.250-2.472, P=0.001). Compared with 3-6BA, the SRB of 3-6BC blastocysts was lower (IVF: OR=0.621, 95% CI: 0.447-0.862, P=0.004; ICSI: OR=0.442, 95% CI: 0.238-0.818, P=0.009). Taking 3-6AB as reference, the SRB of 3-6AA blastocysts was higher (IVF: OR=1.600, 95% CI: 1.307-1.958, P<0.001; ICSI: OR=3.130, 95% CI: 1.964-4.987, P<0.001). Conclusion:The SRB of fresh embryo group is slightly higher than that of frozen-thawed group in single blastocyst transfer cycles. Different fertilization methods can affect SRB, and the SRB in IVF is significantly higher than that in ICSI group. The quality of blastocysts can affect SRB, and blastocysts with higher quality TE have significantly increased SRB, while ICM quality have no significant effect on SRB.