Objective: To develop a smart blood donation service assistant for popularizing donation-related knowledge to blood donors via intelligent Q&A support, thereby enabling precise service delivery. Methods: Based on the operational scenarios of the Zhejiang Provincial Blood Center, the system utilized the open-source Dify platform for agent orchestration, and integrated with the DeepSeek model as the language processing engine to support online real-time interaction. External tools, including the Amap API and MySQL database queries, were encapsulated via the Model Context Protocol (MCP). A professional blood knowledge base for Retrieval-Augmented Generation (RAG) was constructed using the BGE-M3 embedding model. An innovative dual-large language model collaborative verification mechanism was introduced to design the overall framework. The system was deployed privately using Docker containerization, and offline closed-loop optimization was achieved through customized Python scripts. Results: An interactive interface for blood donors was developed by integrating the chatflow Web component from Dify. The intelligent assistant Agent can recommend optimal blood donation sites and navigation routes by invoking the Amap API based on the donor's location. The Blood Donation Knowledge Agent enables timely responses to inquiries, along with reasonable suggestions and reminders. This agent specializes in the field of voluntary blood donation, empowering the assistant to answer doubts and questions for blood donors in the form of intelligent question-and-answer interaction. It also guides users through preliminary self-assessments, helping potential donors identify eligibility issues beforehand, thereby effectively increasing the on-site success rate of blood donation and reducing resource waste. Conclusion: The smart blood donation assistant validates the feasibility of the "Dify+MCP+RAG" technical architecture within the blood transfusion informatization field. The assistant not only improves the service experience for blood donors, but also, ensures the sustainable evolution of the system through its modular design and closed-loop optimization mechanism, thus providing valuable insights for the intelligent transformation of traditional blood donation service systems.

Objective: To explore the longitudinal relationship between bullying victimization and sleep quality among rural boarding junior high school students in Hebei Province, and to investigate the chain mediated role of loneliness and rumination, so as to provide evidence for promoting sleep health in the population. Methods: A baseline survey was conducted in May, 2023 (T1) by convenient sampling method, and two rounds of longitudinal surveys were conducted in November, 2023 (T2) and May, 2024 (T3) among students in two rural boarding junior high schools in Hebei Province, and a sample of 601 students who completed all the surveys was finally obtained. Students completed questionnaires, including the Delaware Bullying Victimization Scale Student (DBVS-S), the University of California at Los Angeles Loneliness Scale (UCLA), the Ruminative Responses Scale, and the Pittsburgh Sleep Quality Index (PSQI). Group differences were examined by using t-test or ANOVA, correlations between variables were analyzed by using Pearson correlation coefficients, and a serial mediation structural equation model was constructed, with mediation effects tested via the Bootstrap method. Results: Female students scored higher on sleep quality than male students (7.47±2.70, 6.47 ±2.46, t =4.74, P <0.01). Pearson correlation analysis indicated that bullying victimization was positively correlated with loneliness, rumination, and sleep quality; loneliness was positively correlated with rumination and sleep quality; and rumination was positively correlated with sleep quality ( r =0.26, 0.33, 0.23; 0.39, 0.38; 0.54, all P <0.01). Mediation analysis showed that T2 loneliness had an independent mediating effect of 0.70 (95% CI =0.36-1.35) between T1 bullying victimization and T3 sleep quality, T2 rumination had an independent mediating effect of 1.34 (95% CI =0.71-2.45), and the serial mediation effect of T2 loneliness and T2 rumination was 0.64 (95% CI =0.37-1.13), accounting for 22.11% of the total effect (all P <0.01). Conclusions Bullying victimization adversely affects sleep quality among rural boarding junior high school students through a longitudinal chain mediating pathway involving loneliness and rumination. Psychological interventions should be strengthened for students who experience bullying to alleviate their loneliness and reduce rumination, thereby improving sleep quality.

Objective: To investigate the influencing factors for kinesiophobia among elderly patients with chronic obstructive pulmonary disease (COPD), so as to provide the reference for alleviating kinesiophobia among COPD patients. Methods: From December 2023 to July 2024, COPD patients aged 60 years and above who sought medical treatment at a tertiary grade-a hospital in Guiyang City were selected. Demographic information was collected through questionnaire surveys. Kinesiophobia, exercise self-efficacy, social support, type D personality and coping styles were assessed using the Chinese version of Tampa Scale for Kinesiophobia, the Chinese version of the Self-Efficacy for Exercise Scale, Social Support Rating Scale, Type D Personality Scale and Chinese version of the Medical Coping Modes Questionnaire, respectively. Factors affecting kinesiophobia among elderly patients with COPD were analyzed using a multiple linear regression model. Results: A total of 300 COPD patients were surveyed, including 238 males (79.33%) and 62 females (20.67%). The majority of patients had a disease duration of less than 5 years, with 130 cases (43.33%). The average kinesiophobia score was (48.01±7.74) points. The average exercise self-efficacy score was (3.39±1.01) points. The average social support score was (34.42±6.76) points. There were 280 patients (93.33%) with type D personality. The average scores of the confrontation, avoidance, and resignation dimensions of coping styles were (17.42±5.00), (13.76±1.91), and (11.81±2.95) points, respectively. Multiple linear regression analysis showed that age (70-<80 years, β'=0.124; ≥80 years, β'=0.205), educational level (primary school and below, β'=0.228; junior high school, β'=0.182), household monthly income per capita (<3 000 yuan, β'=0.234; 3 000~<5 000 yuan, β'=0.165), social support (β'=0.294), type D personality (β'= 0.170), and coping styles (confrontation dimension, β'=-0.140; avoidance dimension, β'=0.154; resignation dimension, β'=0.175) statistically associated with kinesiophobia among elderly patients with COPD. Conclusion Kinesiophobia among elderly patients with COPD is associated with age, educational level, household monthly income per capita, social support, type D personality and coping styles.

OBJECTIVE: To investigate the effectiveness of tibial transverse transport (TTT) in treating Wagner grade 3-4 type 2 diabetic foot ulcers and analyze dynamic changes in immunoglobulin levels. METHODS: The clinical data of 68 patients with Wagner grade 3-4 type 2 diabetic foot ulcers treated with TTT between May 2022 and September 2023 was retrospectively analyzed. The cohort included 49 males and 19 females, aged 44-91 years (mean, 67.3 years), with 40 Wagner grade 3 and 28 grade 4 ulcers. The duration of type 2 diabetes ranged from 5 to 23 years, with an average of 10 years. The number of wound healing cases, healing time, amputation cases, death cases, and complications were observed and recorded. Serum samples were collected at 6 key time points [1 day before TTT and 3 days, 7 days (the first day of upward transverse transfer), 14 days (the first day of downward transverse transfer), 21 days (the first day after the end of transfer), 36 days (the first day after the removal of the transfer device)], and the serum immunoglobulin levels were detected by flow cytometry including immunoglobulin G (IgG), IgA, IgM, IgE, complement C3 (C3), C4, immunoglobulin light chain κ (KAP), immunoglobulin light chain λ (LAM). RESULTS: All the 68 patients were followed up 6 months. Postoperative pin tract infection occurred in 3 cases and incision infection in 2 cases. Amputation occurred in 5 patients (7.4%) at 59-103 days after operation, and 8 patients (11.8%) died at 49-77 days after operation; the wounds of the remaining 55 patients (80.9%) healed in 48-135 days, with an average of 80 days. There was no recurrence of ulcer, peri-osteotomy fracture, or local skin necrosis during follow-up. The serum immunoglobulin levels of 55 patients with wound healing showed that the levels of IgG and IgM decreased significantly on the 3rd and 7th day after operation compared with those before operation ( P<0.05), and gradually returned to the levels before operation after 14 days, and reached the peak on the 36th day. IgA levels continued to decrease with time, and there were significant differences at all time points when compared with those before operation ( P<0.05). The level of IgE significantly decreased at 21 days after operation compared with that before operation ( P<0.05), while it was higher at other time points than that before operation, but the difference was not significant ( P>0.05). The level of C3 showed a clear treatment-related increase, which was significantly higher on the 7th, 14th, and 21st days after operation than that before operation ( P<0.05), and the peak appeared on the 14th day. The change trend of C4 level was basically synchronous with that of C3, but the amplitude was smaller, and the difference was significant at 7 and 14 days after operation compared with that before operation ( P<0.05). There was no significant difference in KAP/LAM between different time points before and after operation ( P>0.05). CONCLUSION TTT can accelerate wound healing, effectively treat diabetic foot ulcer, and reduce amputation rate, and has definite effectiveness. The potential mechanisms of TTT in the treatment of diabetic foot ulcers include the dynamic regulation of IgG, IgA, IgM, and IgE levels to balance the process of inflammation and repair, and the periodic increase of C3 and C4 levels may promote tissue cleaning, angiogenesis, and anti-infection defense.
Humans ; Male ; Female ; Middle Aged ; Aged ; Diabetic Foot/immunology* ; Wound Healing ; Adult ; Retrospective Studies ; Aged, 80 and over ; Treatment Outcome ; Tibia/transplantation* ; Diabetes Mellitus, Type 2/complications* ; Amputation, Surgical ; Immunoglobulins/blood* ; Immunoglobulin G/blood*

Metachromatic leukodystrophy (MLD) is an inherited disease caused by a deficiency of the enzyme arylsulfatase A (ARSA). Lentivirus-modified autologous hematopoietic stem cell gene therapy (HSCGT) has recently been approved for clinical use in pre and early symptomatic children with MLD to increase ARSA activity. Unfortunately, this advanced therapy is not available for most patients with MLD who have progressed to more advanced symptomatic stages at diagnosis. Patients with late-onset juvenile MLD typically present with a slower neurological progression of symptoms and represent a significant burden to the economy and healthcare system, whereas those with early onset infantile MLD die within a few years of symptom onset. We conducted a pilot study to determine the safety and benefit of HSCGT in patients with postsymptomatic juvenile MLD and report preliminary results. The safety profile of HSCGT was favorable in this long-term follow-up over 9 years. The most common adverse events (AEs) within 2 months of HSCGT were related to busulfan conditioning, and all AEs resolved. No HSCGT-related AEs and no evidence of distorted hematopoietic differentiation during long-term follow-up for up to 9.6 years. Importantly, to date, patients have maintained remarkably improved ARSA activity with a stable disease state, including increased Functional Independence Measure (FIM) score and decreased magnetic resonance imaging (MRI) lesion score. This long-term follow-up pilot study suggests that HSCGT is safe and provides clinical benefit to patients with postsymptomatic juvenile MLD.
Humans ; Leukodystrophy, Metachromatic/genetics* ; Pilot Projects ; Genetic Therapy/methods* ; Hematopoietic Stem Cell Transplantation ; Male ; Follow-Up Studies ; Female ; Lentivirus/genetics* ; Child ; Child, Preschool ; Hematopoietic Stem Cells/metabolism* ; Cerebroside-Sulfatase/metabolism* ; Adolescent

Objective:To screen for microRNAs(miRNAs)highly expressed in the serum exosomes(Exo)of esophageal squamous cell carcinoma(ESCC)patients and analyze their relationship with the clinicopathological characteristics of the patients,and to explore the potential of Exo-derived miRNAs as clinical auxiliary diagnostic markers for ESCC.Methods:Serum and relevant clinical data of 50 healthy subjects and 45 newly diagnosed ESCC patients admitted to the Fourth Hospital of Hebei Medical University between December 2021 and June 2023 were collected,serving as the control group and the ESCC group respectively.The Gene Expression Omnibus(GEO)database and qPCR were used to screen and identify the candidate miRNA for increased expression in the serum of ESCC patients-miR-1246.The diagnostic efficacy of serum miR-1246 for ESCC was analyzed by the receiver operating characteristic curve.The relationship between miR-1246 and the clinical feature progression of ESCC patients was analyzed by Logistic regression,and the relationship between miR-1246 and the clinicopathological characteristics of ESCC patients was analyzed by the χ2 test.Exosomes in the serum of the subjects were isolated,purified and characterized for verification.The expression of miR-1246 in Exo was detected by qPCR.ESCC KYSE150 and KYSE30 cells were routinely cultured.mimics-NC and miR-1246 mimics were transfected respectively into KYSE150 cells using Lipofectamine 2000.Inhibitor-NC and miR-1246 inhibitor were transfected into KYSE30 cells,which were respectively denoted as the minics-NC,miR-1246 mimics,inhibitor-NC and miR-1246-inhibitor groups.KYSE150 and KYSE30 cells were treated with Exo derived from KYSE150 cells in the mimics-NC and miR-1246 mimics groups.The proliferation,migration and invasion abilities of cells in each group were detected by the CCK-8 assay,scratch wound healing assay and Transwell chamber assay respectively.The expressions of Exo markers,epithelial-mesenchymal transition-related proteins,TET family methylcytosine dioxygenase 2(TET2)and cell adhesion molecule 1(CADM1)proteins in each group of cells were detected by WB assay.The targeting binding relationship between miR-1246 and TET2 and CADM1 was verified by the dual-luciferase reporter gene assay.Results:Bioinformatics screening showed that the miRNA with the most significant differential expression in the serum of ESCC patients was miR-1246.The serum Exo extracted from the patients conformed to the typical Exo characteristics.The expression level of serum Exo-miR-1246 in ESCC patients at stages Ⅰ-Ⅱ was significantly higher than that in healthy subjects(P<0.01);the level of serum Exo-miR-1246 in ESCC patients at stages Ⅲ-Ⅳ was significantly higher than that in patients at stages Ⅰ-Ⅱ(P<0.01).ROC curve analysis showed that Exo-miR-1246 in serum had a high value for auxiliary differential diagnosis of ESCC(P<0.05),and the auxiliary diagnostic efficacy of Exo-miR-1246 for the clinical progression of ESCC patients was higher than that of CEA and SCC-Ag(P<0.05).The combined detection of the three could further improve the efficacy of auxiliary diagnosis of patient staging(P<0.01).Exo-miR-1246 might be an independent risk factor for the clinical progression of ESCC patients(P<0.05).The expression level of serum Exo-miR-1246 was associated with the T-stage,N-stage and clinical stage of ESCC(P<0.01).Overexpression of miR-1246 could promote the proliferation,migration,invasion,epithelial-mesenchymal transition and inhibit apoptosis of ESCC cells,while inhibition of miR-1246 had the opposite effect.Database data analysis found that TET2 and CADM1 were the target genes of miR-1246.The dual-luciferase reporter gene assay confirmed that miR-1246 could directly bind to TET2 and CADM1 mRNA and inhibit their expressions(P<0.01).Treatment of KYSE150 and KYSE30 cells with Exo derived from cells overexpressing miR-1246 had the same effect as overexpressing miR-1246 in these cells.Conclusion:Exo-derived miR-1246 has the potential to be a clinical auxiliary diagnostic marker for ESCC.It may affect the occurrence and development of ESCC by regulating the expression levels of TET2 and CADM1.

OBJECTIVE To investigate the expression of long non-coding RNA(lncRNA)HOXC13-AS in head and neck squamous cell carcinoma tissues and its effects on tumor cell proliferation and migration in head and neck tumor cell lines.METHODS Clinical samples from 26 patients with laryngeal squamous cell carcinoma and 4 patients with hypopharyngeal squamous cell carcinoma who were treated surgically in the Second Hospital of Jilin University from March 2024 to December 2024 were collected as research subjects.Head and neck squamous cell carcinoma tissue specimens and corresponding adjacent tissue specimens were selected.The expression of HOXC13-AS in 30 cases of head and neck squamous cell carcinoma tissues and corresponding adjacent tissues was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Clinical data were collected to analyze the correlation between HOXC13-AS expression and various clinical pathological factors.siRNA technology was used to interfere with the expression of HOXC13-AS in laryngeal cancer cell line TU212 and hypopharyngeal squamous cell line FaDu.Cell Counting Kit-8(CCK-8)assay was used to detect cell proliferation ability,and cell scratch assay was used to detect cell migration ability.RESULTS The expression level of lncRNA HOXC13-AS in head and neck squamous cell carcinoma tissues(12.60±11.26)was significantly higher than that in corresponding adjacent tissues(1.40±0.61),with a statistically significant difference(t=5.485,P<0.001).The expression of HOXC13-AS in head and neck squamous cell carcinoma tissues was closely related to T stage(P=0.004),lymph node metastasis(P=0.006),and clinical stage(P=0.007)of patients.CCK-8 results showed that the absorbance values of TU212 cell line groups at 24,48,and 72 h were si-NC group(0.373±0.010,0.738±0.026,1.003±0.124),si-HOXC13-AS-1 group(0.365±0.015,0.686±0.019,0.935±0.028),and si-HOXC13-AS-2 group(0.364±0.024,0.700±0.026,0.943±0.053),with statistically significant differences(F24 h=0.484,F48 h=7.893,F72 h=1.345,P<0.01);the absorbance values of FaDu cell line groups at 24,48,and 72 h were si-NC group(0.727±0.054,0.834±0.072,1.224±0.127),si-HOXC13-AS-1 group(0.532±0.005,0.650±0.079,1.021±0.044),and si-HOXC13-AS-2 group(0.647±0.088,0.687±0.025,1.074±0.055),with statistically significant differences(F24 h=16.143,F48 h=14.259,F72 h=9.409,P<0.001).Cell scratch assay showed that the migration rates of TU212 cell line groups at 12 and 24 h were si-NC group(38.971%±3.824%,69.185%±0.469%),si-HOXC13-AS-1 group(18.182%±2.580%,33.378%±2.302%),and si-HOXC13-AS-2 group(25.017%±0.288%,48.413%±0.805%),with statistically significant differences(F12 h=47.295,F24 h=471.745,P<0.0001);The migration rates of FaDu cell lines in each group at 12 and 24 hours were as follows:si-NC group(39.067%±3.196%,58.222%±0.448%),si-HOXC13-AS-1 group(13.689%±0.132%,39.358%±3.985%),and si-HOXC13-AS-2 group(23.335%±0.680%,35.526%±0.758%),with statistically significant differences(F12 h=138.1,F24 h=101.749,P<0.0001).CONCLUSION HOXC13-AS is significantly upregulated in head and neck squamous cell carcinoma tissues and cells,and is associated with the clinical pathological characteristics of patients.Silencing HOXC13-AS can significantly inhibit the proliferation and migration of head and neck tumor cells.

Objective To retrieve,evaluate,and integrate evidence related to the operational proce-dures of ambulatory blood pressure monitoring(ABPM)in adults,aiming to enhance the accuracy and effec-tiveness of ambulatory blood pressure monitoring.Methods A systematic search was conducted following the"6S"pyramid model of evidence-based resources to identify literature pertaining to ABPM operations in adults from relevant domestic and international databases and websites,with the search period spanning from the inception of each database to April 2024.After screening the literature,the methodological quality of the included studies was evaluated.Evidence was extracted and summarized according to thematic categories.Results Based on the inclusion and exclusion criteria,15 publications were ultimately included,comprising 1 clinical decision,10 guidelines,1 best practice summary,and 3 expert consensus documents.A total of 32 evidence items were synthesized.Conclusions This study consolidates evidence related to ABPM operational procedures,providing an evidence-based foundation for standardizing ABPM practices among healthcare pro-fessionals.

Objective:To screen for microRNAs(miRNAs)highly expressed in the serum exosomes(Exo)of esophageal squamous cell carcinoma(ESCC)patients and analyze their relationship with the clinicopathological characteristics of the patients,and to explore the potential of Exo-derived miRNAs as clinical auxiliary diagnostic markers for ESCC.Methods:Serum and relevant clinical data of 50 healthy subjects and 45 newly diagnosed ESCC patients admitted to the Fourth Hospital of Hebei Medical University between December 2021 and June 2023 were collected,serving as the control group and the ESCC group respectively.The Gene Expression Omnibus(GEO)database and qPCR were used to screen and identify the candidate miRNA for increased expression in the serum of ESCC patients-miR-1246.The diagnostic efficacy of serum miR-1246 for ESCC was analyzed by the receiver operating characteristic curve.The relationship between miR-1246 and the clinical feature progression of ESCC patients was analyzed by Logistic regression,and the relationship between miR-1246 and the clinicopathological characteristics of ESCC patients was analyzed by the χ2 test.Exosomes in the serum of the subjects were isolated,purified and characterized for verification.The expression of miR-1246 in Exo was detected by qPCR.ESCC KYSE150 and KYSE30 cells were routinely cultured.mimics-NC and miR-1246 mimics were transfected respectively into KYSE150 cells using Lipofectamine 2000.Inhibitor-NC and miR-1246 inhibitor were transfected into KYSE30 cells,which were respectively denoted as the minics-NC,miR-1246 mimics,inhibitor-NC and miR-1246-inhibitor groups.KYSE150 and KYSE30 cells were treated with Exo derived from KYSE150 cells in the mimics-NC and miR-1246 mimics groups.The proliferation,migration and invasion abilities of cells in each group were detected by the CCK-8 assay,scratch wound healing assay and Transwell chamber assay respectively.The expressions of Exo markers,epithelial-mesenchymal transition-related proteins,TET family methylcytosine dioxygenase 2(TET2)and cell adhesion molecule 1(CADM1)proteins in each group of cells were detected by WB assay.The targeting binding relationship between miR-1246 and TET2 and CADM1 was verified by the dual-luciferase reporter gene assay.Results:Bioinformatics screening showed that the miRNA with the most significant differential expression in the serum of ESCC patients was miR-1246.The serum Exo extracted from the patients conformed to the typical Exo characteristics.The expression level of serum Exo-miR-1246 in ESCC patients at stages Ⅰ-Ⅱ was significantly higher than that in healthy subjects(P<0.01);the level of serum Exo-miR-1246 in ESCC patients at stages Ⅲ-Ⅳ was significantly higher than that in patients at stages Ⅰ-Ⅱ(P<0.01).ROC curve analysis showed that Exo-miR-1246 in serum had a high value for auxiliary differential diagnosis of ESCC(P<0.05),and the auxiliary diagnostic efficacy of Exo-miR-1246 for the clinical progression of ESCC patients was higher than that of CEA and SCC-Ag(P<0.05).The combined detection of the three could further improve the efficacy of auxiliary diagnosis of patient staging(P<0.01).Exo-miR-1246 might be an independent risk factor for the clinical progression of ESCC patients(P<0.05).The expression level of serum Exo-miR-1246 was associated with the T-stage,N-stage and clinical stage of ESCC(P<0.01).Overexpression of miR-1246 could promote the proliferation,migration,invasion,epithelial-mesenchymal transition and inhibit apoptosis of ESCC cells,while inhibition of miR-1246 had the opposite effect.Database data analysis found that TET2 and CADM1 were the target genes of miR-1246.The dual-luciferase reporter gene assay confirmed that miR-1246 could directly bind to TET2 and CADM1 mRNA and inhibit their expressions(P<0.01).Treatment of KYSE150 and KYSE30 cells with Exo derived from cells overexpressing miR-1246 had the same effect as overexpressing miR-1246 in these cells.Conclusion:Exo-derived miR-1246 has the potential to be a clinical auxiliary diagnostic marker for ESCC.It may affect the occurrence and development of ESCC by regulating the expression levels of TET2 and CADM1.

To retrieve, evaluate, and integrate evidence related to the operational procedures of ambulatory blood pressure monitoring (ABPM) in adults, aiming to enhance the accuracy and effectiveness of ambulatory blood pressure monitoring.