BACKGROUND:The mesenchymal stem cell secretome contains bioactive substances,cytokines,and growth factors.Three-dimensional cell culture can regulate the secretion of these components,potentially enhancing the ability to promote injury repair.OBJECTIVE:To investigate the repair effect of three-dimensional cultured human umbilical cord mesenchymal stem cell secretome on skin injuries in mice.METHODS:Human umbilical cord mesenchymal stem cells were cultured in conventional two-dimensional culture dishes and 96-well U-bottom cell culture plates,from which their secretory components were subsequently collected.The expression of skin damage repair related secretory factors in umbilical cord mesenchymal stem cells was analyzed using RT-qPCR.The protein expression level of skin damage repair related factors in umbilical cord mesenchymal stem cell secretome was detected using enzyme-linked immunosorbent assay.The potential of human umbilical cord mesenchymal stem cell secretome to repair vascular injuries was evaluated using an immortalized human umbilical vein endothelial cell migration model.A mouse skin injury model was established,and the human umbilical cord mesenchymal stem cell secretome was injected subcutaneously.Repair effects on skin injury were assessed through wound healing rates and histopathological analysis.RESULTS AND CONCLUSION:(1)After three days of cultivation,human umbilical cord mesenchymal stem cells cultured in two dimensions exhibited a fibroblast-like,swirling growth pattern,whereas three-dimensional culture led to the formation of uniform microspheres.(2)Compared with two-dimensional culture,three-dimensional culture significantly increased the mRNA expression of transforming growth factor β and basic fibroblast growth factor in human umbilical cord mesenchymal stem cells.(3)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly enhanced the protein expression of vascular endothelial growth factor,interleukin-10,and granulocyte-macrophage colony-stimulating factor in the human umbilical cord mesenchymal stem cell secretome.(4)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly promoted the migration of immortalized human umbilical cord mesenchymal stem cells.(5)Compared with the untreated control group and the two-dimensional cultured human umbilical cord mesenchymal stem cell secretome,the three-dimensional cultured human umbilical cord mesenchymal stem cell secretome can significantly accelerate the skin wound healing rate and wound skin structure remodeling in mice.These results indicate that three-dimensional culture can enhance the expression of paracrine factors of human umbilical cord mesenchymal stem cells,and their secretome can significantly promote the repair of mouse skin damage.

BACKGROUND:The mesenchymal stem cell secretome contains bioactive substances,cytokines,and growth factors.Three-dimensional cell culture can regulate the secretion of these components,potentially enhancing the ability to promote injury repair.OBJECTIVE:To investigate the repair effect of three-dimensional cultured human umbilical cord mesenchymal stem cell secretome on skin injuries in mice.METHODS:Human umbilical cord mesenchymal stem cells were cultured in conventional two-dimensional culture dishes and 96-well U-bottom cell culture plates,from which their secretory components were subsequently collected.The expression of skin damage repair related secretory factors in umbilical cord mesenchymal stem cells was analyzed using RT-qPCR.The protein expression level of skin damage repair related factors in umbilical cord mesenchymal stem cell secretome was detected using enzyme-linked immunosorbent assay.The potential of human umbilical cord mesenchymal stem cell secretome to repair vascular injuries was evaluated using an immortalized human umbilical vein endothelial cell migration model.A mouse skin injury model was established,and the human umbilical cord mesenchymal stem cell secretome was injected subcutaneously.Repair effects on skin injury were assessed through wound healing rates and histopathological analysis.RESULTS AND CONCLUSION:(1)After three days of cultivation,human umbilical cord mesenchymal stem cells cultured in two dimensions exhibited a fibroblast-like,swirling growth pattern,whereas three-dimensional culture led to the formation of uniform microspheres.(2)Compared with two-dimensional culture,three-dimensional culture significantly increased the mRNA expression of transforming growth factor β and basic fibroblast growth factor in human umbilical cord mesenchymal stem cells.(3)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly enhanced the protein expression of vascular endothelial growth factor,interleukin-10,and granulocyte-macrophage colony-stimulating factor in the human umbilical cord mesenchymal stem cell secretome.(4)Compared with two-dimensional culture,three-dimensional cultured human umbilical cord mesenchymal stem cell secretome significantly promoted the migration of immortalized human umbilical cord mesenchymal stem cells.(5)Compared with the untreated control group and the two-dimensional cultured human umbilical cord mesenchymal stem cell secretome,the three-dimensional cultured human umbilical cord mesenchymal stem cell secretome can significantly accelerate the skin wound healing rate and wound skin structure remodeling in mice.These results indicate that three-dimensional culture can enhance the expression of paracrine factors of human umbilical cord mesenchymal stem cells,and their secretome can significantly promote the repair of mouse skin damage.

[摘 要] 目的：探讨lncRNA DLEU2通过EZH2/H3K27me3途径调控IKKα介导甲状腺癌（TC）放射性碘抵抗的作用机制。方法：利用TCGA数据库分析TC中DLEU2的表达及其与EZH2的相关性。构建放射性碘抵抗的TPC-1细胞（RR-TPC-1细胞）模型及裸鼠移植瘤模型，通过敲低或过表达DLEU2（si-DLEU2/OE-DLEU2）、抑制EZH2（UNC1999）、过表达IKKα（OE-IKKα）进行干预，采用qPCR、WB、RIP、ChIP、CCK-8、流式细胞术、TUNEL染色及体内成瘤实验检测基因与蛋白表达、表观修饰、细胞增殖、凋亡及肿瘤生长。结果：TCGA分析显示，DLEU2在TC组织中显著上调（P < 0.001），与患者不良预后相关（P = 0.008 4），且与EZH2表达呈正相关（r = 0.390, P < 0.001）；RIP证实EZH2与DLEU2存在相互作用/结合（P < 0.05）。体外实验表明，敲低DLEU2可显著下调RR-TPC-1细胞中EZH2、IKKα表达及H3K27me3修饰水平，抑制NF-κB通路活化（P < 0.05或P < 0.01），抑制细胞增殖、促进凋亡（均P < 0.05）。联合敲低DLEU2与抑制EZH2进一步增强上述效应，而过表达IKKα则可部分逆转上述效应（P < 0.05或P < 0.01）。体内实验进一步证实，敲低DLEU2联合抑制EZH2可显著抑制移植瘤生长，增加肿瘤细胞凋亡（均P < 0.01）；IKKα过表达则部分逆转上述抗肿瘤效应（P < 0.05或P < 0.01）。结论：lncRNA DLEU2通过招募EZH2催化H3K27me3修饰，间接激活IKKα/NF-κB信号并形成正反馈环路，介导TPC-1细胞131I抵抗。

Objective To understand the antibody levels of diphtheria and tetanus among healthy population in Shanghai Pudong New Area, and to provide a scientific basis for improving the vaccine immunization strategy. Methods Random sampling was used to select healthy people of all ages in 16 communities in Shanghai Pudong New Area from 2018 to 2024, and serum samples were collected and tested for serum anti-diphtheria and tetanus toxin IgG antibodies by enzyme-linked immunosorbent assay (ELISA) method to analyze the antibody positivity rate (≥0.1 IU/ml) and the geometric mean concentration (GMC) of antibodies. Results A total of 3 312 serum samples were included, with a male-to-female ratio of 0.76:1, and 53.77% were local residents. The seropositivity rates and geometric mean concentrations (GMC) of both diphtheria and tetanus antibodies generally declined with increasing age, but exhibited a transient rebound in the 7y-. A total of 1 175 individuals (35.48%) were seropositive for diphtheria, with a GMC of 0.054 IU/mL. For tetanus, 988 individuals (29.83%) were seropositive, with a GMC of 0.033 IU/mL. Significant differences in seropositivity rates (χ²_diphtheria=950.005,χ²_tetanus=1 324.393) and GMC (H_diphtheria=1027.160,H_tetanus=1 142.007) were observed among different age groups (P<0.001). Significant differences in seropositivity rates (χ²_diphtheria=950.005,χ²_tetanus=1324.393) and GMC (H_diphtheria=1027.160,H_tetanus=1142.007) were also found across different years (P<0.001). Conclusion The prevalence of diphtheria and tetanus antibodies in the healthy population of Pudong New Area is relatively low, particularly among adults over 20 years of age with inadequate immunization. This underscores the need to reinforce the National Immunization Program (NIP) vaccine specifications for children under 6 years of age and implement an immunization strategy for adolescents or adults against diphtheria and tetanus.

Objective: To explore the effects of a classroom based transdiagnostic preventive intervention program for emotional disorders on depressive and anxiety symptoms, and resilience among junior high school students, so as to provide reference for promoting adolescent mental health. Methods: From October to December 2023, 72 students from a junior high school in Beijing were recruited as research subjects and divided into an intervention group ( n =36) and a control group ( n =36). The intervention group participated in 8 sessions (40 mins per session) of transdiagnostic prevention intervention course for emotional disorders; the control group students participated in research based learning lessons or class meetings. Patient Health Questionnaire-9 (PHQ-9), Generalized Anxiety Disorder-7 (GAD-7), and Resilience Scale-8 (RS-8) were used to evaluate junior high school students. The Wilcoxon signed rank test was used to compare the differences in depressive and anxiety symptoms, and resilience scores between the intervention group and the control group before and after the intervention. PROCESS macro was used to test the mediating effect. Results: There were no statistically significant differences in the scores of depressive and anxiety symptoms, and resilience before and after intervention in the intervention group ( Z =-1.07, 0.07, 1.43, all P >0.05). In the control group, the depressive symptoms score was higher after the intervention than before, and the resilience score was lower than before the intervention ( Z = 2.23 , -2.53, both P <0.05). After intervention, the depressive symptoms score in the intervention group was lower than that in the control group, and the resilience score in the intervention group was higher than that in the control group ( Z =-2.09, 3.33, both P < 0.05). The increase in the scores of resilience played a complete mediating role between the intervention and depressive symptoms, with the mediating effect value being -0.98 (95% CI =-2.47 to -0.11), accounting for 30.98% of the total effect. Conclusion Transdiagnostic preventive intervention program for emotional disorders based on classroom could prevent the deterioration of depressive symptom in junior high school students, and maintaining the level of resilience is an important factor for the intervention effect.

Objective To explore the method for establishing a pig left lung orthotopic transplantation model. Methods A total of 4 male Yorkshire pigs weighing (40.0±2.5) kg underwent surgical procedures, including anesthesia, tracheal intubation, donor lung procurement, and recipient lung transplantation. Histological morphology and blood gas analysis of the transplanted lungs were assessed 2 hours after reperfusion to evaluate the histological changes and function of the transplanted lungs. Results The pigs underwent left lung in situ transplantation, effectively simulating conditions relevant to human lung transplantation. Two hours after the transplantation, arterial blood gas analysis showed that arterial partial pressure of oxygen was 155.4-178.6 mm Hg, arterial partial pressure of carbon dioxide was 53.1-62.4 mm Hg, and the oxygenation index was 310.8-357.2 mm Hg. Hematoxylin and eosin staining indicated a low degree of pulmonary edema and minimal cellular infiltration. Conclusion The pig left lung orthotopic transplantation model possesses strong operability and stability. Researchers can replicate this model according to the described methods and further conduct basic research and explore clinical translational applications.

BackgroundObstructive sleep apnea （OSA） represents a potential modifiable risk factor for cognitive impairment， while spatial navigation performance serves as a sensitive marker to detect such deficits at an early stage. Currently， polysomnography （PSG） is the gold standard for OSA diagnosis， with the apnea hypopnea index （AHI） used to stratify the disease severity. Nevertheless， AHI alone cannot fully explain the individual variability in cognitive impairment among OSA patients. Existing evidence indicates that sleep architecture disruption impairs spatial memory consolidation and subsequently alters spatial navigation through pathways independent of AHI， yet this hypothesis remains unvalidated in routine clinical OSA cohorts. ObjectiveTo compare the global cognitive function and spatial navigation performance across OSA patients of varying severities， and to analyze associated factors of spatial navigation performance， so as to provide clinical evidence for the early identification and intervention of cognitive impairment in OSA patients. MethodsThis cross-sectional study consecutively enrolled 135 outpatients from the Mental Health Center of Inner Mongolia Autonomous Region from March 8， 2024 to September 12， 2025. All participants completed PSG monitoring and fulfilled the OSA diagnostic criteria outlined in the Guidelines for the Diagnosis and Treatment of Adult Obstructive Sleep Apnea. Subjects were stratified into mild （5 ≤AHI < 15 events/h）， moderate （15 ≤ AHI < 30 events/h）， and severe （AHI ≥ 30 events/h） groups based on AHI values， and all PSG sleep architecture parameters were extracted for statistical analysis. Global cognitive function was evaluated using the Mini-Mental State Examination （MMSE）， the Trail Making Test （including TMT-A and TMT-B）， and two subtasks from the THINC Integrated Toolkit （THINC-it）： the Spotter test （choice reaction time， CRT） and the Codebreaker test （digit symbol substitution test， DSST）. Spatial navigation performance was assessed through the Santa Barbara Sense of Direction Scale （SBSOD） and the computerized AMUNET spatial navigation test. ResultsAmong the 135 OSA patients， 46 （34.07%） were classified into the mild group， 47 （34.82%） into the moderate group， and 42 （31.11%） into the severe group. In terms of cognitive function， there were no statistically significant intergroup differences across three groups in CRT， consecutive correct DSST responses， MMSE scores， and completion time of TMT-A and TMT-B （P>0.05）. Regarding spatial navigation performance， no statistically significant differences were observed among three groups in SBSOD scores and the outcomes of the four AMUNET subtasks （P>0.05）. Spearman correlation analyses revealed that the performance on the delayed allocentric AMUNET subtask was positively correlated with stage N2 duration， percentage of stage N2 sleep （N2%）， and rapid eye movement （REM） latency （r=0.277， 0.296， 0.239， P_FDR<0.05）. In contrast， this subtask exhibited negative correlations with REM duration， percentage of REM sleep （REM%）， stage N3 duration and percentage of stage N3 sleep （N3%） （r=-0.223–-0.175， P_FDR<0.05）. SBSOD score was positively correlated with the completion time of TMT-A and TMT-B （r=0.236， 0.278）， and negatively correlated with MMSE scores （r=-0.311）. For AMUNET indices， performance on the allocentric navigation subtask showed a positive correlation with CRT （r=0.013）. Moreover， performance of both egocentric and allocentric navigation subtasks yielded positive correlations with the completion time of TMT-A and TMT-B （r=0.191–0.265， P_FDR<0.05）. Performance on the egocentric， allocentric， and combined egocentric–allocentric navigation subtasks was negatively correlated with MMSE scores and the consecutive correct DSST responses （r=-0.318–-0.207， P_FDR<0.05）. Hierarchical multiple liner regression analysis demonstrated that after controlling for baseline characteristics and global cognitive function， N2% emerged as a positive predictor of the delayed allocentric navigation subtask performance （β=0.242， 95% CI： 0.104–1.463）. ConclusionPatients with OSA across all severity strata exhibit comparable global cognitive function and spatial navigation performance. Spatial memory is associated with sleep architecture parameters， among which N2% acts as an independent positive predictor of spatial memory performance. ［Funded by Natural Science Foundation Project of Inner Mongolia Autonomous Region （number， 2024QN08050）； Science and Technology Project for High-Level Clinical Specialty Development of Public Hospitals of the Capital Region under the Autonomous Region Health Commission of Inner Mongolia Autonomous Region in 2023 （number， 2023SGGZ047）］

With the rapid advancement of medical technology and the increasing complexity of clinical environments, competence in clinical ethical decision-making has become an indispensable core competency for exemplary physicians. However, traditional medical ethics education has long been confronted with the persistent challenges of theory-practice dissociation and the disconnect between pedagogical settings and clinical realities. Thus, how to construct a "Clinical Ethics" curriculum that aligns with the objectives of training high-caliber medical professionals has emerged as a critical and challenging issue in current medical education reform. To effectively address this challenge, the Office of Education at Peking University Third Hospital, since 2021, has assembled a multidisciplinary "Clinical Ethics" teaching and management team comprising faculty from Peking University School of Humanities and key clinical specialists from the hospital. In 2022, the "Clinical Ethics" elective course was formally launched for fourth-year students in the eight-year MD program who had completed coursework in "Medical Ethics" and organ-system-based curricula, with the aim of exploring and operationalizing a pre-clinical ethics education model tailored for senior medical students. The course employs a modular instructional design encompassing "theoretical review-case-based deliberation-clinical workshop", which effectively facilitates the deep integration of medical humanities with clinical practice.Following four years of iterative refinement through the "plan-act-observe-reflect" cycle, the course has demonstrated positive outcomes in enhancing students' ethical awareness and decision-making capabilities, and has preliminarily established a sustainable collaborative mechanism among the multidisciplinary faculty team. This article systematically delineates the course development background, pedagogical model and methodological innovations, and the multidisciplinary collaborative framework, alongside reflections and summaries based on implementation outcomes, with the intent of providing references for the future development of "Clinical Ethics" curricula.

Objective : To investigate the status of electronic cigarette use and its influencing factors among residents aged ≥15 years in Chengdu City in 2021 and 2024, so as to provide the evidence for optimizing electronic cigarette control strategies. Methods: Two rounds of cross-sectional surveys were conducted in 2021 and 2024. Multi-stage cluster random sampling method was used to select permanent residents aged ≥15 years in Chengdu City as survey subjects. The Chinese Adult Tobacco Survey Questionnaire was used to collect information on gender, age, and electronic cigarette use. Complex sampling weighting was used to calculate the rate of electronic cigarette use and the proportion of reasons for electronic cigarette use. A complex sampling multivariable logistic regression model was used to analyze the influencing factors for electronic cigarette use. Results: A total of 7 950 and 6 920 people were surveyed in 2021 and 2024, respectively, including 3 868 and 3 653 males (48.65% and 52.79%), and 4 082 and 3 267 females (51.35% and 47.21%), respectively. The rate of electronic cigarette use in 2024 was lower than in 2021 (2.29% vs. 3.15%, P<0.05). In 2024, the rates of electronic cigarette use were lower in males, rural areas, people aged 15-<45 years, educational level of bachelor' s degree or above, occupation in government/public institutions or occupation in enterprises/commercial/service industries, and current smokers compared with 2021, while the rate of electronic cigarette use was higher in residents aged ≥65 years compared with 2021 (all P<0.05). In 2021 and 2024, the main reasons for electronic cigarette use were smoking cessation and the belief that electronic cigarettes are less harmful. Smoking cessation accounted for 27.78% and 23.28%, and the belief that electronic cigarettes are less harmful accounted for 24.26% and 22.10%, respectively. Complex sampling multivariable logistic regression analysis showed that age (15-<25 years, OR=15.682, 95%CI: 5.660-43.450; 25-<45 years, OR=7.463, 95%CI: 2.987-18.647; 45-<65 years, OR=3.425, 95%CI: 1.562-7.510), educational level (bachelor' s degree or above, OR=2.916, 95%CI: 1.444-5.891), and smoking (current, OR=23.992, 95%CI: 8.182-70.353; former, OR=20.393, 95%CI: 6.150-67.624) were influencing factors for electronic cigarette use in 2021. Gender (male, OR=2.279, 95%CI: 1.074-4.832), age (25-<45 years, OR=2.114, 95%CI: 1.148-3.892), educational level (high school/vocational high school/technical secondary school, OR=2.792, 95%CI: 1.414-5.514; bachelor' s degree or above, OR=3.321, 95%CI: 1.567-7.039), occupation (unemployed, retired, and retired, OR=0.449, 95%CI: 0.226-0.892), and smoking (current, OR=10.547, 95%CI: 5.563-19.994; former, OR=8.998, 95%CI: 3.833-21.125) were influencing factors for electronic cigarette use in 2024. Conclusions Compared with 2021, the rate of electronic cigarette use among residents in Chengdu City decreased in 2024. Young people, males, bachelor' s degree or above, and current smokers are the key populations for electronic cigarette control. The cognitive bias held by residents that electronic cigarette can aid in smoking cessation and pose minimal harm needs to be corrected.

ObjectiveTo investigate the clinical efficacy and safety of state-target formula Angelica and Safflower spot removal essence in the treatment of melasma. MethodsA randomized，double-blind，placebo-controlled parallel-group trial was conducted，selecting patients with Qi stagnation and blood stasis type of melasma who visited the dermatology outpatient department of Beijing Hospital of Integrated Traditional Chinese and Western Medicine from September 2022 to February 2023. In accordance with the double-blind principle，a random number table was generated to randomly divide the patients into an experimental group （50 cases） and a control group （50 cases）. The experimental group applied the Angelica and Safflower spot removal essence topically，while the control group applied a placebo topically，with continuous treatment for 8 weeks and a follow-up of 1 month. Melasma Area and Severity Index （MASI），Physician Global Assessment （PGA），patient self-assessment scoring，VISIA detection and scoring were performed before medication and on the first day after stopping medication，and all adverse events were recorded. ResultsIn a study involving 100 patients，87 cases were included in the analysis. The experimental group consisted of 45 cases，with 5 dropouts，and the control group had 42 cases，with 8 dropouts. The baseline data of the patients in both groups were consistent，with no statistically significant differences. After treatment，the total effective rate of the experimental group was 75.56% （34/45），significantly higher than the control group's 2.38% （1/42），and the difference was statistically significant （χ²=48.38，P<0.01）. Compared with before treatment，after treatment，the MASI score，VISIA image spot，brown spot，and red area score of the experimental group patients were significantly reduced （P<0.05，P<0.01），while there was no statistically significant difference in the control group patients. Compared with the control group after treatment，the experimental group showed more significant improvement （P<0.05，P<0.01）. VISIA image analysis showed that after 8 weeks of treatment，the experimental group showed significant improvement in surface spots，deep spots，and red areas，while the control group remained unchanged or showed an increasing trend. 42.22% （19/45） of the experimental group had a PGA score of 3 or below，while 88.89% （40/45） had a PGA score of 4 or below. 80.95% （34/42） of the control group had a PGA score of 5 or above，and the difference between the two groups was statistically significant （χ²=38.26，P<0.01）. There was no statistically significant difference in the recurrence rate and adverse reaction rate within one month between the experimental group and the control group. ConclusionThe state-target formula Angelica and Safflower spot removal essence is clinically effective and safe in the treatment of melasma.