1.Application advances, ethical dilemmas, and future directions of large language models in lung cancer diagnosis and treatment
Zhizhen REN ; Yufan XI ; Xu ZHU ; Yijie LUO ; Geting HUANG ; Junqiao SONG ; Xiuyuan XU ; Nan CHEN ; Qiang PU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(03):353-362
Lung cancer is a leading cause of cancer-related morbidity and mortality worldwide. Coupled with the substantial workload, the clinical management of lung cancer is challenged by the critical need to efficiently and accurately process increasingly complex medical information. In recent years, large language models (LLMs) technology has undergone explosive development, demonstrating unique advantages in handling complex medical data by leveraging its powerful natural language processing capabilities, and its application value in the field of lung cancer diagnosis and treatment is continuously increasing. The paper systematically analyzes that the exceptional potential of LLMs in lung cancer auxiliary diagnosis, tumor feature extraction, automatic staging, progression/outcome analysis, treatment recommendations, medical documentation generation, and patient education. However, they face critical technical and ethical challenges including inconsistent performance in complex integrated decision-making (e.g., TNM staging, personalized treatment suggestions) and "black box" opacity issues, along with dilemmas such as training data biases, model hallucinations, data privacy concerns, and cross-lingual adaptation challenges ("data colonization"). Future directions should prioritize constructing high-quality multimodal corpora specific to lung cancer, developing interpretable and compliant specialized models, and achieving seamless integration with existing clinical workflows. Through dual drivers of technological innovation and ethical standardization, LLMs should be prudently advanced for holistic lung cancer management processes, ultimately promoting efficient, standardized, and personalized diagnosis and treatment practices.
2.Mendelian randomization study on the association between telomere length and 10 common musculoskeletal diseases
Weidong LUO ; Bin PU ; Peng GU ; Feng HUANG ; Xiaohui ZHENG ; Fuhong CHEN
Chinese Journal of Tissue Engineering Research 2025;29(3):654-660
BACKGROUND:Multiple observational studies have suggested a potential association between telomere length and musculoskeletal diseases.However,the underlying mechanisms remain unclear. OBJECTIVE:To investigate the genetic causal relationship between telomere length and musculoskeletal diseases using two-sample Mendelian randomization analysis. METHODS:Genome-wide association study summary data of telomere length were obtained from the UK Biobank.Genome-wide association study summary data of 10 common musculoskeletal diseases(osteonecrosis,osteomyelitis,osteoporosis,rheumatoid arthritis,low back pain,spinal stenosis,gout,scapulohumeral periarthritis,ankylosing spondylitis and deep venous thrombosis of lower limbs)were obtained from the FinnGen consortium.Inverse variance weighting,Mendelian randomization-Egger and weighted median methods were used to evaluate the causal relationship between telomere length and 10 musculoskeletal diseases.Inverse variance weighting was the primary Mendelian randomization analysis method,and sensitivity analysis was performed to explore the robustness of the results. RESULTS AND CONCLUSION:(1)Inverse variance-weighted results indicated a negative causal relationship between genetically predicted telomere length and rheumatoid arthritis(odds ratio=0.78,95%confidence interval:0.64-0.95,P=0.015)and osteonecrosis(odds ratio=0.56,95%confidence interval:0.36-0.90,P=0.016).No causal relationship was found between telomere length and the other eight musculoskeletal diseases(all P>0.05).(2)Sensitivity analysis affirmed the robustness of these causal relationships,and Mendelian randomization-Egger intercept analysis found no evidence of potential horizontal pleiotropy(all P>0.05).(3)This Mendelian randomized study supports that telomere length has protective effects against rheumatoid arthritis and osteonecrosis.However,more basic and clinical research will be needed to support our findings in the future.
3.Complications among patients undergoing orthopedic surgery after infection with the SARS-CoV-2 Omicron strain and a preliminary nomogram for predicting patient outcomes.
Liang ZHANG ; Wen-Long GOU ; Ke-Yu LUO ; Jun ZHU ; Yi-Bo GAN ; Xiang YIN ; Jun-Gang PU ; Huai-Jian JIN ; Xian-Qing ZHANG ; Wan-Fei WU ; Zi-Ming WANG ; Yao-Yao LIU ; Yang LI ; Peng LIU
Chinese Journal of Traumatology 2025;28(6):445-453
PURPOSE:
The rate of complications among patients undergoing surgery has increased due to infection with SARS-CoV-2 and other variants of concern. However, Omicron has shown decreased pathogenicity, raising questions about the risk of postoperative complications among patients who are infected with this variant. This study aimed to investigate complications and related factors among patients with recent Omicron infection prior to undergoing orthopedic surgery.
METHODS:
A historical control study was conducted. Data were collected from all patients who underwent surgery during 2 distinct periods: (1) between Dec 12, 2022 and Jan 31, 2023 (COVID-19 positive group), (2) between Dec 12, 2021 and Jan 31, 2022 (COVID-19 negative control group). The patients were at least 18 years old. Patients who received conservative treatment after admission or had high-risk diseases or special circumstances (use of anticoagulants before surgery) were excluded from the study. The study outcomes were the total complication rate and related factors. Binary logistic regression analysis was used to identify related factors, and odds ratio (OR) and 95% confidence interval (CI) were calculated to assess the impact of COVID-19 infection on complications.
RESULTS:
In the analysis, a total of 847 patients who underwent surgery were included, with 275 of these patients testing positive for COVID-19 and 572 testing negative. The COVID-19-positive group had a significantly higher rate of total complications (11.27%) than the control group (4.90%, p < 0.001). After adjusting for relevant factors, the OR was 3.08 (95% CI: 1.45-6.53). Patients who were diagnosed with COVID-19 at 3-4 weeks (OR = 0.20 (95% CI: 0.06-0.59), p = 0.005), 5-6 weeks (OR = 0.16 (95% CI: 0.04-0.59), p = 0.010), or ≥7 weeks (OR = 0.26 (95% CI: 0.06-1.02), p = 0.069) prior to surgery had a lower risk of complications than those who were diagnosed at 0-2 weeks prior to surgery. Seven factors (age, indications for surgery, time of operation, time of COVID-19 diagnosis prior to surgery, C-reactive protein levels, alanine transaminase levels, and aspartate aminotransferase levels) were found to be associated with complications; thus, these factors were used to create a nomogram.
CONCLUSION
Omicron continues to be a significant factor in the incidence of postoperative complications among patients undergoing orthopedic surgery. By identifying the factors associated with these complications, we can determine the optimal surgical timing, provide more accurate prognostic information, and offer appropriate consultation for orthopedic surgery patients who have been infected with Omicron.
Humans
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COVID-19/complications*
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Male
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Female
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Middle Aged
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Postoperative Complications/epidemiology*
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SARS-CoV-2
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Orthopedic Procedures/adverse effects*
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Aged
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Nomograms
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Adult
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Retrospective Studies
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Risk Factors
4.Exploration on Mechanism of Topical Treatment of Allergic Contact Dermatitis in Mice with Portulacae Herba Based on Nrf2/HO-1/NF-κB Signaling Pathway
Xiaoxue WANG ; Guanwei FAN ; Xiang PU ; Zhongzhao ZHANG ; Xia CHEN ; Ying TANG ; Nana WU ; Jiangli LUO ; Xiangyan KONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):115-123
ObjectiveTo investigate the mechanism of topical treatment of allergic contact dermatitis (ACD) mice with Portulacae Herba based on the nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1)/nuclear factor-κB (NF-κB) signaling pathway. MethodsA total of 70 6-week-old specific pathogen free (SPF) female Kunming mice were adaptively fed for 1 week and randomly divided into blank group, model group, compound dexamethasone acetate cream group (2.075×10-2 g·g-1), blank matrix cream group, low-dose Portulacae Herba cream group (0.1 g·g-1), high-dose Portulacae Herba cream group (0.2 g·g-1), and Portulacae Herba + inhibitor group (0.2 g·g-1 + 30 mg·kg-1 ML385), with 10 mice in each group. One day before the experiment, the mice were shaved on the neck and back. Except for the blank group, the mice in the other groups were treated with 2,4-dinitrochlorobenzene (DNCB) to establish an ACD model. After respective administration, the skin lesion of the mice was scored, and the histopathological changes of the skin were stained with hematoxylin-eosin (HE). Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), reactive oxygen species (ROS), superoxide dismutase (SOD) activity, and malondialdehyde (MDA) in serum of mice. The expression of Nrf2/HO-1/NF-κB signaling pathway-related proteins in mouse skin tissue was detected by immunohistochemistry (IHC), Western blot, and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultsCompared with the blank group, the mice in the model group had an increased skin lesion score (P<0.01), severe pathological damage to skin tissue, increased content of IL-1β, IL-6, ROS, and MDA in their serum (P<0.01), and decreased content of SOD (P<0.01). In addition, the mRNA and protein expression levels of Nrf2, HO-1, and nuclear factor-κB inhibitor α (IκBα) in skin tissue were up-regulated (P<0.01), while the protein expression levels of phosphorylated (p)-IκBα and p-NF-κB p65 and the mRNA expression of NF-κB p65 were down-regulated (P<0.01). Compared with the model group and the blank matrix cream group, the mice treated with Portulacae Herba had a decreased skin lesion score (P<0.01), reduced pathological damage to skin tissue, decreased content of IL-1β, IL-6, ROS, and MDA in their serum (P<0.01), and increased content of SOD (P<0.01). Additionally, the mRNA and protein expression levels of Nrf2, HO-1, and IκBα in skin tissue were down-regulated (P<0.05,P<0.01), and the protein expression levels of p-IκBα and p-NF-κB p65 and the mRNA expression of NF-κB p65 were up-regulated (P<0.05,P<0.01). Compared with the Portulacae Herba + inhibitor group, the high-dose Portulacae Herba cream group had a decreased skin lesion score (P<0.01), alleviated pathological damage to skin tissue, decreased content of IL-1β, IL-6, ROS, and MDA in the serum of mice (P<0.05,P<0.01), and increased content of SOD (P<0.01). The protein expression levels of Nrf2, HO-1, and IκBα and the mRNA expression of Nrf2 and HO-1 in skin tissue were up-regulated (P<0.05,P<0.01), and the protein expression levels of p-IκBα and p-NF-κB p65 and the mRNA expression of NF-κB p65 were down-regulated (P<0.05). ConclusionPortulacae Herba can improve DNCB-induced ACD skin damage in mice by regulating the Nrf2/HO-1/NF-κB signaling pathway.
5.Efficacy of cannulated screw internal fixation combined with quadratus femoris bone flap with preservation of the posterior superior retinaculum for femoral neck fracture in young and middle-aged patients
Huan LUO ; Tianhua ZHOU ; Chuan LI ; Luqiao PU ; Xingbo CAI ; Teng WANG ; Chen MENG ; Yaolin ZHANG ; Yongqing XU
Chinese Journal of Trauma 2025;41(1):65-71
Objective:To compare the efficacy of cannulated screw internal fixation combined with quadratus femoris bone flap with preservation of the posterior superior retinaculum and cannulated screw internal fixation alone in the treatment of femoral neck fracture in young and middle-aged patients.Methods:A retrospective cohort study was conducted to analyze the clinical data of 83 young and middle-aged patients with femoral neck fracture admitted to the 920th Hospital of Joint Logistic Support Force of PLA from January 2018 to January 2023, including 56 males and 27 females, aged 28-55 years [(42.7±3.2)years]. According to Garden classification, the fractures were classified as type III in 22 patients and type IV in 61. Based on Pauwels classification, the fractures were classified as type I in 15 patients, type II in 38 and type III in 30. Forty patients were treated with cannulated screw internal fixation combined with modified quadratus femoris bone flap (cannulated screw combined with bone flap group) and 43 with cannulated screw internal fixation alone (cannulated screw group). The two groups were compared in terms of the operation time, intraoperative blood loss, time to weight-bearing, length of hospital stay, and wound healing. The visual analogue scale (VAS) scores and Harris hip function scores at 1, 3, 6, 12 months after surgery and at the last follow-up. The postoperative complication rate was detected.Results:All the patients were followed up for 20-70 months [(40.0±1.2)months]. The operation time and intraoperative blood loss were (105.2±2.7)minutes and (100.6±16.3)ml in the cannulated screw combined with bone flap group, which were longer or more than (92.4±4.7)minutes and (92.5±14.6)ml in the cannulated screw group ( P<0.01). The time to weight-bearing was (12.1±1.4)weeks in the cannulated screw combined with bone flap group, shorter than (23.6±1.2)weeks in the cannulated screw group ( P<0.01). There was no statistically significant difference in the length of hospital stay between the two groups (P>0.05). The incisions in both groups were healed by first intention. At 1 month after surgery, no statistically significant difference was observed in VAS scores between the two groups ( P>0.05); at 3, 6, 12 months after surgery and at the last follow-up, the VAS scores were (6.6±0.2)points, (4.5±0.3)points, (3.2±0.5)points, and (2.6±0.4)points in the cannulated screw combined with bone flap group, lower than (7.0±0.1)points, (5.2±0.2)points, (3.9±0.4)points, and (3.3±0.1)points in the cannulated screw group ( P<0.05 or 0.01). At 1 and 3 months after surgery, no statistically significant difference was observed in the Harris hip function scores between the two groups ( P>0.05); at 6, 12 months after surgery and at the last follow-up, the Harris hip function scores were (82.2±1.7)points, (90.0±1.4)points, and (91.6±1.0)points in the cannulated screw combined with bone flap group, higher than (75.2±1.7)points, (83.4±1.9)points, and (85.2±0.7)points in the cannulated screw group ( P<0.01). At the last follow-up, in the cannulated screw combined with bone flap group, the Harris hip function was rated excellent in 32 patients, good in 5, and fair in 3, with an excellent and good rate of 92.5%, while in the cannulated screw group, the Harris hip function was rated excellent in 20 patients, good in 13, and fair in 10, with an excellent and good rate of 76.7% ( P<0.05). The postoperative complication rate was 5.0% (2/40) in the cannulated screw combined with bone flap group, significantly lower than 23.2% (10/43) in the cannulated screw group ( P<0.05). Conclusion:Compared with cannulated screw internal fixation alone, cannulated screw internal fixation combined with quadratus femoris bone flap with preservation of the posterior superior retinaculum has the advantages of earlier weight-bearing, less pain, better recovery of hip joint function, and lower incidence of postoperative complications in the treatment of femoral neck fracture in young and middle-aged patients, despite longer operation time and more intraoperative blood loss.
6.Research progress on the application of Traditional Chinese Medicine-related technologies in brain protection for aerospace crew members
Xinyi SHENG ; Lianghui MENG ; Yan LUO ; Yuyang PU ; Shirong WANG ; Ziqi FANG ; Rong LIANG ; Jiajia YANG
Space Medicine & Medical Engineering 2025;36(5):479-485
In recent years,with the continuous advancement of deep space exploration missions,astronauts have been increasingly exposed to complex and extreme environmental factors in space,such as microgravity,space radiation,circadian rhythm disruption,and confined isolation.These conditions can easily induce brain dysfunction in astronauts,manifesting as cognitive decline,emotional disturbances,sleep disorders,and neuroinflammatory responses.Traditional Chinese Medicine(TCM),characterized by its holistic regulation and syndrome differentiation-based treatment principles,exhibits multi-target regulation and systemic coordination,and has attracted growing attention in the field of brain function protection.This study systematically reviews the protective applications and research progress of TCM and related techniques in both actual spaceflight missions and simulated space environment models against brain functional disorders.The underlying mechanisms and future prospects are discussed,with the aim of providing insights for safeguarding astronauts'brain health and laying a theoretical foundation for the precise intervention and systematic protection strategies of TCM in future deep space missions.
7.Effects of Portulaca oleracea cream on skin pruritus and barrier function in allergic contact dermatitis mice
Xiaoxue WANG ; Xia CHEN ; Xiang PU ; Guanwei FAN ; Xiangyan KONG ; Ying TANG ; Nana WU ; Jiangli LUO
China Pharmacy 2025;36(11):1352-1357
OBJECTIVE To study the effects and mechanism of Portulaca oleracea cream on skin pruritus and barrier function in allergic contact dermatitis (ACD) mice. METHODS Low-concentration and high-concentration P. oleracea creams were prepared, with the P. oleracea extract solution (1 g/mL, calculated by crude drug) concentrations of 10% and 20%. Sixty BALB/c mice were randomly allocated into blank group, model group, Mometasone furoate cream group (positive control), blank matrix cream group, P. oleracea low-concentration and high-concentration cream groups. Except for blank group, ACD model was induced in each group using 2,4-dinitrochlorobenzene solution. The blank group and model groups received normal saline, while the remaining groups were treated with their respective creams, once a day, at a dose of approximately 0.5 g per application, continuously for 14 days. At 24 h post-final administration, skin lesions of mice were observed and scored; pathological changes of skin tissue were observed; serum levels of immunoglobulin E(IgE) and tumor necrosis factor-α (TNF-α) were quantified. mRNA expression of MAS-related G protein-coupled receptors (including MrgprA3, MrgprC11, and MrgprD) in dorsal root ganglion (DRG) was assessed; while protein expressions of skin barrier function-related proteins Claudin-1 and Occludin in skin tissues were determined. RESULTS Compared with blank group, mice in the model group exhibited severe skin damage, characterized by loss of epidermal architecture, hyperkeratosis of the skin tissue, and the infiltration of a large number of inflammatory cells. The skin injury scores, as well as the serum levels of IgE and TNF-α, and the mRNA expression levels of MrgprA3, MrgprC11, and MrgprD in DRG, were all significantly elevated compared to the blank group (P<0.05 or P<0.01); in contrast, the protein expression levels of Claudin-1 and Occludin in the skin tissue were markedly reduced (P<0.05). Compared with model group, mice in P. oleracea low-concentration and high- concentration cream groups demonstrated significant alleviation of skin damage, as evidenced by reduced epidermal hyperplasia, mitigated spongiosis in the dermis, and decreased infiltration of inflammatory cells; these quantitative indicators were almost significantly reversed (P<0.05 or P<0.01). No significant differences were observed in the aforementioned skin injuries, pathological alterations, or quantitative indicators between the blank matrix cream group and the model group. CONCLUSIONS P. oleracea may ameliorate skin lesions and restore skin barrier function of ACD mice, the mechanism of which may be associated with downregulating mRNA expressions of MrgprA3, MrgprC11 and MrgprD in DRG, and up-regulating the protein expressions of Claudin-1 and Occludin in skin tissue.
8.Preparation of polyphenol-mediated copper ion coating on titanium surface and antibacterial and antioxidant properties
Zhenju GUAN ; Yonglin XIE ; Shougang XIANG ; Chengdong ZHANG ; Xiaolong LI ; Xingping LI ; Chao PU ; Bo ZHANG ; Xuwei LUO ; Dongqin XIAO
Chinese Journal of Tissue Engineering Research 2025;29(10):1997-2005
BACKGROUND:Titanium implants are widely used in clinical practice because of their high strength and good biocompatibility.However,during implantation,bacterial infection and tissue damage environment produce a large number of reactive oxygen species,which can easily lead to delayed tissue healing and surgical failure.Consequently,the development of titanium implants with antimicrobial and antioxidant properties becomes paramount. OBJECTIVE:Considering the potent antimicrobial attributes of copper ions and the remarkable antioxidant qualities of polyphenols,we proposed the fabrication of polyphenol-mediated copper ion coatings on titanium surfaces.These coatings were subsequently assessed for their in vitro antimicrobial and antioxidant properties. METHODS:Nanostructures were generated on the titanium surface using the alkali thermal method.The titanium was immersed in a solution containing tannic acid and copper ions to achieve polyphenol-mediated copper ion coatings.The surface morphology and water contact angle were detected.The loading and release of copper ions were examined using atomic absorption spectroscopy.Staphylococcus aureus was inoculated on the surface of pure titanium sheet(blank group),alkali heat treated titanium sheet(control group),and polyphenol mediated copper ion modified titanium sheet(experimental group)to observe the bacterial survival status.Osteoblast precursor cells MC3T3-E1 were co-cultivated on the surface of three groups of titanium sheets to assess their antioxidant properties and bioactivity. RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that the polyphenol-mediated copper ion modified titanium sheet had rod-like nanostructures and no cracks on the surface.The surface hydrophilicity of copper ion modified titanium sheet mediated by polyphenol was close to that of pure titanium sheet.Atomic absorption spectrometry results showed a 51%increase in the loading capacity of copper ions after polyphenol mediation,with a uniform release of copper ions.(2)The antibacterial rates of titanium sheets in the blank group,control group,and experimental group were 0%,21.65%,and 93.75%,respectively.The live/dead staining and CTC staining showed that the live bacteria on the surface of titanium plates in the blank group were the most,and the live bacteria on the surface of titanium plates in the experimental group were the least.(3)The results of live/dead staining and CCK-8 assay showed that the three groups of titanium sheets had good cytocompatibility,and the titanium sheets in the experimental group were more conducive to the proliferation of MC3T3-E1 cells.Active oxygen fluorescence probe detection exhibited that compared with the other two groups,the fluorescence intensity of active oxygen on the surface of the experimental group was significantly reduced.The results of alkaline phosphatase and alizarin red S staining showed that the osteogenic differentiation and extracellular matrix mineralization of MC3T3-E1 cells on the surface of titanium sheets in the experimental group were stronger than those in the other two groups.(4)These results show that the polyphenol-mediated copper ion coating has strong antibacterial and antioxidant properties and promotes osteogenic differentiation.
9.Analysis of the influencing factors of health-related quality of life in community-dwelling elderly with mild cognitive impairment from the perspective of health ecology
Jiayi LIN ; Yanbo ZHU ; Jiameng JIA ; Yuhao LUO ; Jiaju REN ; Jianni CONG ; Yueheng LOU ; Liqun LONG ; Rui CAO ; Pu GE
Chinese Journal of Behavioral Medicine and Brain Science 2025;34(3):245-251
Objective:To analyze the current situation of health-related quality of life (HRQOL) and its influencing factors among community-dwelling elderly with mild cognitive impairment (MCI) based on the health ecology model (HEM).Methods:From December 2023 to September 2024, a cross-sectional survey was conducted among 997 community-dwelling elderly in Beijing, Shandong, Chongqing and other regions. The EuroQol five-dimensional questionnaire 5 level (EQ-5D-5L), the Montreal cognitive assessment (MoCA), and the Pittsburgh sleep quality index (PSQI) were used to assess the subjects.Descriptive analysis, Mann-Whitney U test, Kruskal-Wallis test, and Tobit regression analysis were performed using Stata 18.0 software. Results:A total of 324 valid samples were finally obtained, and the health utility value of MCI elderly was 0.842(0.815, 0.951). Tobit regression analysis showed that ≥80 years old( β=-0.121, 95% CI=-0.193--0.050) in individual traits level, don't exercise( β=-0.196, 95% CI=-0.255--0.137) and poor quality of sleep ( β=-0.064, 95% CI=-0.116--0.013) in behavior characteristics level, and poor subjective economic status( β=-0.153, 95% CI=-0.261--0.045) in living and working conditions level were risk factors for the health utility value of the elderly with MCI. Drinking( β=0.096, 95% CI=0.022-0.171) in behavior characteristics level and participating in social activities( β=0.126, 95% CI=0.062-0.190) in interpersonal network level were protective factors for the health utility value of the elderly with MCI. Conclusions:The HRQOL of the community-dwelling elderly with MCI in China is low, and its influencing factors are multi-level. HEM should be combined to strengthen the intervention and management of the elderly with MCI from personal constitution to policy environment to improve their HRQOL.
10.Iron overload induces ferroptosis in osteoblast precursor cells and inhibits osteogenic differentiation
Yu PAN ; Renfeng ZHAO ; Xingping LI ; Chengdong ZHANG ; Feng SHI ; Chao PU ; Xuwei LUO ; Dongqin XIAO
Chinese Journal of Tissue Engineering Research 2025;29(30):6381-6390
BACKGROUND:Iron overload is an independent factor inducing osteoporosis,but the action mechanism is currently unclear.Therefore,exploring the effects of iron overload on osteoblast-related cells will help to deeply understand the pathogenesis of osteoporosis and provide potential strategies for osteoporosis treatment.OBJECTIVE:To explore the effects of iron overload environment on osteoblast precursor cell activity,ferroptosis,and osteogenic differentiation.METHODS:Osteoblast precursor cells(MC3T3-E1 cells)were divided into blank group,iron overload group,fer-1 group,and deferoxamine group.The iron overload group was treated with 300 μmol/L ammonium ferric citrate in the culture medium for 48 hours to simulate the iron overload microenvironment.The cells in fer-1 group and deferoxamine group were pretreated with 5 μmol/L antioxidant fer-1 and 5 μmol/L deferoxamine for 8 hours,respectively,and then added with 300 μmol/L ammonium ferric citrate for 48 hours.CCK-8 assay was used to determine the cell viability.Intracellular reactive oxygen species levels were detected employing a reactive oxygen species fluorescent probe.Changes in mitochondrial membrane potential were monitored with a mitochondrial membrane potential fluorescent probe.Mitochondrial morphology was observed employing transmission electron microscopy.Cellular glutathione levels were measured with a reduced glutathione colorimetric assay kit.Lipid peroxidation levels were assessed with a malondialdehyde colorimetric assay kit.Cellular ferrous ion levels were determined with a ferrous ion colorimetric assay kit.The osteogenic and mineralization capabilities of the cells were verified by alkaline phosphatase staining and alizarin red staining.Collagen secretion ability was detected using Sirius Red staining.The expression of osteogenic/ferroptosis-related genes and proteins was examined through reverse transcription quantitative polymerase chain reaction and western blot analysis.RESULTS AND CONCLUSION:(1)In an iron-overload environment,the mitochondrial membrane potential of cells decreased and their structure was compromised,with an elevation in intracellular lipid peroxidation levels and a downregulation of genes and proteins associated with ferroptosis resistance.However,pretreatment with fer-1 and deferoxamine led to an increase in mitochondrial membrane potential and partial restoration of morphology,a reduction in intracellular lipid peroxidation levels,and an upregulation of genes and proteins related to ferroptosis resistance.(2)In an iron-overload environment,the levels of cellular alkaline phosphatase,the formation of mineralized nodules,and the synthesis of collagen fibers were all found to be decreased.Pretreatment with fer-1 and deferoxamine was observed to upregulate the expression of osteogenic differentiation in cells.(3)In summary,iron overload could increase intracellular oxidative stress levels,mediate ferroptosis in MC3T3-E1 cells and inhibit osteogenic differentiation,thereby inducing osteoporosis.Therefore,maintaining iron homeostasis and inhibiting osteogenesis-related ferroptosis may be potential strategies to prevent or treat osteoporosis.

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