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.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.
3.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.
4.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*
;
Male
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Female
;
Middle Aged
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Postoperative Complications/epidemiology*
;
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
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.Protective effects of icariin against radiation-induced cardiac disease in mice
Fengmin YIN ; Chaoyuan PU ; Tao RAN ; Zixuan SU ; Mengjia WU ; Lei ZHANG ; Xinyi LUO ; Qilin LIU ; Yan CHEN ; Qihai GONG ; Wei HU
Chinese Journal of Radiological Medicine and Protection 2025;45(2):83-90
Objective:To explore the cardioprotective effects of icariin (ICA) against radiation-induced cardiac disease (RICD) in C57BL/6 mice.Methods:A total of 48 female C57BL/6J mice aged 6-8 weeks were randomly divided into three groups: the control group (CON), the irradiation group (IR), and the irradiation combined with icariin group (IR+ ICA), with 16 mice in each group. The IR and IR+ ICA groups received a single cardiac irradiation at a dose of 30 Gy, while the CON group received no radiation treatment. The IR+ ICA group was treated with ICA (70 mg·kg -1·d -1) two weeks before irradiation until the end of the experiment through intragastric administration. In contrast, the CON and IR groups were treated with an equal volume of vehicle solution (0.5% sodium carboxymethyl cellulose, NaCMC) via intragastric administration. The mice′s mental status, food intake, body weight, and survival rates were monitored during the experiment. At two weeks post-irradiation, the venous blood of the mice was collected and serum was separated for the enzyme-linked immunosorbent assays (ELISA) of creatine kinase MB isoenzyme (CK-MB) and cardiac troponin T (cTnT/TNNT2). At 12 weeks post-irradiation, the cardiac function of the mice was assessed using echocardiography. After the mice were euthanized under anesthesia, the histopathological changes and fibrosis degree of their myocardial tissues were assessed using hematoxylin and eosin (HE) and Masson′s trichrome staining, followed by the calculation of collagen volume fraction (CVF). The differential gene expression of brain natriuretic peptide (BNP), transforming growth factor-β (TGF-β), and interleukin-6 (IL-6) in the cardiac tissues of the mice was detected using real-time reverse transcription-polymerase chain reaction (RT-PCR). Apoptosis-related proteins and proteins associated with the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway were determined using Western blotting. The survival curves of the mice were plotted using Kaplan-Meier, and the survival differences of the mice among various groups were compared using the log-rank test. Results:After irradiation, the mice in the IR group showed lethargy, as well as decreased food intake and activity, while these symptoms in the IR+ ICA group were significantly alleviated. At two weeks post-irradiation, the CK-MB and cTnT levels of the IR group were significantly elevated compared with the CON group ( t = 5.28, 8.89, P < 0.01). At 12 weeks post-irradiation, the mice in the IR group exhibited significantly decreased body weight ( t = 2.47, P < 0.05) and decreased survival rates ( HR = 8.25, 95% CI: 1.157-58.770, P < 0.05) compared with the CON group. Echocardiography revealed that the IR group featured decreased left ventricular ejection fraction (EF), decreased fractional shortening (FS), and increased left ventricular end-diastolic diameter (LVDD) compared with the CON group ( t = 7.02, 4.45, P < 0.05). Histopathological examination revealed that the IR group suffered from cardiomyocyte edema, disordered arrangement, and increased fibrosis, with an elevated CVF. The IR group exhibited significantly upregulated gene expression of BNP, TGF-β, and IL-6 in cardiac tissues compared with the CON group ( t = 4.23, 6.39, 4.61, P < 0.05). After-irradiation, the IR group exhibited upregulated apoptosis-related proteins Cleaved Caspase-3 and Bax ( t = 6.29, 9.54, P < 0.05), decreased Bcl-2 expression ( t = 8.20, P < 0.001), and decreased phosphorylation levels of PI3K and Akt ( t = 6.47, 3.42, P < 0.001). The symptoms of the mice were partially ameliorated after treatment with ICA. Specifically, the mice in the IR+ ICA group exhibited higher body weight ( t = 5.13, P < 0.001) and significantly higher survival rates ( HR = 0.121, 95% CI: 0.017-0.864, P < 0.05) than the IR group. Compared to the IR group, the IR+ ICA group showed elevated cardiac function indicators EF and FS( t = 3.23, 3.05, P < 0.05), and reduced LVDD ( t = 3.02, P < 0.05). The histopathological analysis revealed mitigated edema and disordered arrangement of cardiomyocytes in the IR+ ICA group. Furthermore, the IR+ ICA group exhibited significantly lower BNP, TGF-β, and IL-6 expression levels than the IR group ( t = 2.83, 4.15, 2.96, P < 0.05). The expression of apoptosis-related proteins Cleaved Caspase-3 and Bax was lower ( t = 3.23, 3.24, P < 0.05), Bcl-2 expression was higher ( t = 5.92, P < 0.001), and restored phosphorylation levels of PI3K and Akt ( t = 2.89, 8.35, P < 0.001). Conclusions:Icariin has protective effects against the RICD. It alleviates cardiomyocyte apoptosis possibly by upregulating the phosphorylation levels of PI3K and Akt.
7.Influence of pre-radiotherapy ultrasonic monitoring of bladder filling levels on setup errors in cervical cancer patients
Jiangyan LUO ; Haizhen YUE ; Jiacheng LIU ; Yichen PU ; Zihong LU ; Jianqi HU ; Hao WU
Chinese Journal of Radiological Medicine and Protection 2025;45(4):290-295
Objective:To investigate the influence of ultrasonic monitoring of bladder filling levels on setup errors before fractionated radiotherapy for cervical cancer patients through a comparative analysis, and its effectiveness in improving clinical target volume (CTV) margins.Methods:A retrospective study was conducted on 1 284 error data of setup via cone beam CT (CBCT) and 6D setup error correction system from 172 cervical cancer patients treated in the Radiotherapy Department of Peking University Cancer Hospital from January 2019 to October 2023. These patients were classified into two groups: 87 (659 times of setup) with ultrasonic monitoring of bladder filling levels and 85 (625 times of setup) without ultrasonic monitoring. The setup errors, error distributions, and numbers of abnormal setups between the two groups were compared in the lateral (Lat), longitudinal (Lng), vertical (Vrt), pitch (Pitch), roll (Roll), and rotational (Rtn) dimensions. Moreover, the CTV to planning target volume(PTV) margin values in the three-dimensional direction were calculated for both groups to assess the clinical value of ultrasonic monitoring of bladder filling levels before fractionated radiotherapy.Results:Compared to the group without ultrasonic monitoring, the group with ultrasonic monitoring exhibited lower median values of setup errors in all six-dimensional directions and smaller upper and lower interquartile ranges ( Z = -10.86 to -6.34, P<0.05). The group with ultrasonic monitoring manifested more concentrated setup errors in various directions and statistically significantly reduced numbers of abnormal setups ( χ2=15.33, P<0.05). Moreover, CTV-PTV margins of the group with ultrasonic monitoring displayed reduced CTV-PTV margin values by 0.55, 1.52, and 1.26 mm in the Vrt, Lng, and Lat directions, respectively. Conclusions:Pre-radiotherapy ultrasonic monitoring of bladder filling levels in cervical cancer patients can significantly improve the repeatability of setup, thus notably reducing the incidence of abnormal setups. Theoretically, it can narrow the range from the CTV to the PTV, thereby minimizing radiation exposure to healthy tissues and ultimately enhancing radiotherapy precision for cervical cancer and reducing radiation damage.
8.Sinicization and psychometric properties of adolescent personality structure questionnaire among Chinese adolescents
Ling ZHANG ; Meng LI ; Yufang ZHAO ; Keyu PU ; Jie LUO
Chinese Journal of Behavioral Medicine and Brain Science 2025;34(11):1043-1050
Objective:To sinicize the adolescent personality structure questionnaire(APS-Q), and to examine the psychometric properties of APS-Q among Chinese adolescents.Methods:APS-Q was sinicized and the Chinese version of APS-Q was used for investigation.A sample of 1 207 adolescents were investigated firstly and randomly divided into sample 1 ( n1=590) and sample 2 ( n2=617) for exploratory factor analysis and confirmatory factor analysis, respectively. Moreover, the sample 3 ( n3=420) was collected to conduct confirmatory factor analysis for the second time.Sample 4( n4=83) was used to assess the test-retest reliability of APS-Q. The Chinese big five personality inventory-15 (CBF-PI-15), generalized anxiety disorder-7 (GAD-7) and patient health questionnaire-9 (PHQ-9) were used as criterion tools. Both SPSS 21.0 and Mplus 7.4 software were used for the data analysis. Results:Item analysis showed that the CITCs for items 27, 31 and 35 were low.Exploratory factor analysis suggested that after deleting items 3, 4, 21, 22, 24, 25, 26, 27, 31 and 35, the revised APS-Q was consisted with 25 items, covering 6 dimensions, which were investment and goals, aggression, sense of self, relationship with family, relationship with friends, and self-acceptance, respectively. Confirmatory factor analysis supported the six-factor model fitted well (sample 2: χ2/ df=1.94, RMSEA=0.04, CFI=0.94, TLI=0.93, and SRMR=0.05; sample 3: χ2/ df=2.12, RMSEA=0.05, CFI=0.95, TLI=0.92, and SRMR=0.06). Multigroup CFA indicated that the six-factor structure of APS-Q had strict measurement invariance across sex. The scores of APS-Q were significantly correlated with big five personality, depression, and anxiety(all P<0.05). The total score of APS-Q demonstrated satisfactory internal consistency (α=0.90, ω=0.90, split-half reliability=0.82, MIC=0.27) and test-retest reliability ( r=0.84), while the subscale scores of APS-Q showed acceptable internal consistencies (α=0.71-0.82, ω=0.72-0.82, split-half reliabilities=0.60-0.79, MIC=0.36-0.52) and test-retest coefficients ( r=0.66-0.77). Conclusion:The Chinese version of APS-Q can be used as a reliable and valid measurement to assess personality function for Chinese adolescents.
9.Impact of lymph node dissection extent on survival in intrahepatic cholangiocarcinoma at different anatomical sites
Weixuan XIE ; Yang BAI ; Huaisheng XU ; Yifeng PU ; Lin WANG ; Zheng FANG ; Qingzhou ZHU ; Kunlun LUO
Chinese Journal of General Surgery 2025;34(8):1680-1687
Background and Aims:The incidence of intrahepatic cholangiocarcinoma(ICC)has been increasing in recent years.Due to its insidious onset and low rate of early diagnosis,radical resection remains the only potential curative treatment.Lymph node metastasis is a major adverse prognostic factor in ICC,but the scope and therapeutic value of lymphadenectomy remain controversial.Previous studies suggest that patients with central ICC may derive greater survival benefit from lymphadenectomy than those with peripheral ICC.Preoperative assessment of lymph node status mainly relies on imaging,but its accuracy is limited.This study aimed to investigate the prognostic impact of lymphadenectomy in ICC patients at different anatomical sites to inform surgical decision-making.Methods:A retrospective analysis was conducted on 220 ICC patients who underwent radical resection at the 904th Hospital of the Joint Logistic Support Force of the PLA from May 2016 to May 2021.The cohort included 126 males and 94 females,with a mean age of(56.76±13.15)years.Patients were categorized into peripheral ICC(n=144)and central ICC(n=76)groups.Clinical characteristics,albumin-bilirubin(ALBI)grade,preoperative risk of lymph node metastasis,number of lymph nodes dissected,lymph node metastasis status,and postoperative survival outcomes were compared.Subgroup analyses were conducted to assess the prognostic value of the number of lymph nodes dissected under different risk stratifications.Results:Significant differences were observed between peripheral and central ICC in ALBI grade(x2=9.952,P=0.002),preoperative lymph node metastasis risk(x2=6.166,P=0.014),number of lymph nodes dissected(x2=4.167,P=0.042),and lymph node metastasis rate(x2=7.331,P=0.007).The 3-year overall survival(OS)rate was higher in peripheral ICC(31.94%)than in central ICC(15.79%)(x2=13.890,P<0.001).Among central ICC patients,those with ≥6 lymph nodes dissected had better 3-year OS than those with<6(16.89%vs.13.04%,x2=3.894,P=0.048).In the high-risk subgroup of central ICC,≥6 lymph nodes dissected was also associated with improved 3-year OS compared with<6(15.62%vs.11.11%,x2=3.962,P=0.047).In contrast,the number of lymph node dissections had no significant prognostic impact in peripheral ICC or in patients classified as low risk.Conclusion:Patients with peripheral ICC had a better prognosis than those with central ICC.Adequate lymphadenectomy(≥6 nodes)improved survival and enhanced staging accuracy in central ICC patients at high risk of lymph node metastasis,highlighting the importance of preoperative risk assessment for optimizing surgical strategies.
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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