1.Establishment of an in vitro model for Erastin and RSL3-induced ferroptosis in hepatocellular carcinoma
Xinyue Zhu ; Qiaohui Ren ; Yan Zang ; Xinyi Zhou ; Junxiao Yao ; Lianzi Wang ; Xudong Sha ; Tao Li
Acta Universitatis Medicinalis Anhui 2025;60(4):611-618
Objective:
To establish anin vitromodel of ferroptosis induced by Erastin and RAS-selective lethal 3(RSL3) in hepatoma cells, and to provide theoretical basis for the development of novel therapeutic strategies for HCC.
Methods:
Hepatoma cells(HCCLM3, HepG2, Hep3B, Huh7 and PLC/PRF/5) in logarithmic growth phase were treated with Erastin(0-40 μmol/L) and RSL3(0-10 μmol/L) at double concentrations respectively. After 24 h, CCK-8 method was used to detect cell viability, draw growth curve, calculate IC50, and HCC cells sensitive to inducers were selected for follow-up experiments. The effect of inducer on the state of hepatoma cells was observed under light microscope, and immunoblotting and flow cytometry were used to verify whether the ferroptotic modelin vitrowas successfully constructed.
Results:
Huh7, Hep3B and HepG2 cells were sensitive to Erastin and RSL3, but HCCLM3 and PLC/PRF/5 were insensitive to Erastin and RSL3. When the concentration of Erastin and RSL3 reached the maximum, the survival rate was still above 65%. Huh7, Hep3B and HepG2 cells were selected for subsequent experiments. Compared with the control group, the expression of Glutathione peroxidase 4(GPX4), a ferroptotic marker, was down-regulated in a concentration-dependent manner. In Huh7, Hep3B and HepG2 cells, lipid reactive oxygen species(ROS) levels significantly increased after 24 h treatment with 10 μmol/L and 20 μmol/L Erastin, respectively; in Huh7 cells, lipid ROS levels significantly increased after 24 h treatment with 0.5 μmol/L and 1 μmol/L RSL3, respectively; in Hep3B and HepG2 cells, lipid ROS levels significantly increased after 24 h treatment with 1 μmol/L and 2 μmol/L RSL3, respectively, compared with control group.
Conclusion
Huh7, Hep3B and HepG2 cells are highly sensitive to Erastin and RSL3. Huh7, Hep3B and HepG2 cells treated with 10 μmol/L Erastin for 24 h are good models for simulating ferroptosis induced by Erastinin vitro, Huh7 cells treated with 0.5 μmol/L RSL3 for 24 h and Hep3B and HepG2 cells treated with 1 μmol/L RSL3 for 24 h are good models for simulating ferroptosis induced by RSL3in vitro.
2.Legal liability and ethical challenges of artificial intelligence in medical decision making:analysis based on diagnosis and treatment scenarios
Yuanming WANG ; Junxiao GE ; Qingsong XU
Academic Journal of Naval Medical University 2025;46(8):977-981
When artificial intelligence is integrated into medical decision-making,the ambiguous definition of technological autonomy and the multi-party game of responsibility allocation lead to legal disputes over rights and responsibilities as well as ethical challenges.Focusing on clinical diagnosis,treatment strategy generation,and prognosis assessment,this paper reveals the data bias and decision traceability deficiencies caused by the"black box"of algorithms,and then proposes solutions from 3 dimensions:defining responsible parties,standardizing technical defect assessments,and promoting interdisciplinary collaboration.From a legal perspective,based on the"high-risk system"classification under the European Union Artificial Intelligence Act and no-fault liability principle under the product liability relative laws,a 3-tier responsibility framework of developer-operator-medical institution is constructed:for auxiliary systems,operating physicians bear ultimate responsibility;for decision-making systems,developers are required to provide verifiable algorithms and take joint liability.Ethically,dynamic review mechanisms(such as mandatory disclosure of decision tree logic and data balance metrics)are suggested to balance technological efficiency with patients' right to informed consent.
3.Elesclomol combined with Cu2 + induced cuproptosis in hepatoma cell lines PLC/PRF/5 and Huh-7
Qiaohui Ren ; Xinyue Zhu ; Wei Lv ; Yan Zang ; Lianzi Wang ; Xinyi Zhou ; Junxiao Yao ; Tao Li
Acta Universitatis Medicinalis Anhui 2025;60(8):1470-1477
Objective:
To investigate the inhibitory effect of Elesclomol (ES) + Cu2 + on the proliferation of human hepatoma cell lines PLC/PRF/5 and Huh-7 and its potential to induce Cuproptosis.
Methods:
Human hepatoma cell lines PLC/PRF/5 and Huh_7 cells were Cultured in vitro. ES solution , Cu2 + solution and copper chelating agent ammonium tetrathiomolybdate VI (ATTM) solution was treated separately or in combination. The effect of ES + Cu2 + on the survival rate of human hepatoma cell lines PLC/PRF/5 and Huh_7 cells and the effect of ES + Cu2 + on the survival rate after pretreatment with copper chelating agent ATTM were evaluated using CCK_8 kit. The cell death induced by ES + Cu2 + was detected by flow cytometry and the changes of ES + Cu2 + after pretreatment with copper chelating agent ATTM. The expression of Cuproptosis related proteins ATPase copper transporting beta (ATP7B) ,ferredoxin 1 (FDX1) , dihydrolipoamide s_acetyltransferase(DLAT) and superoxide dismutase 1 (SOD1) were detected by Western blot. The effect of ES + Cu2 + on cell proliferation and the reverse effect after ATTM pretreatment was detected by cell scratch assay.
Results:
The toxicity of ES + Cu2 + to human hepatocellular carcinoma cell lines PLC/PRF/5 and Huh_7 was significantly dose_dependent (P < 0. 05) . Compared with the control group , the combined application of ES and Cu2 + had a more significant inhibitory effect on hepatocellular carcinoma cells than ES or Cu2 + alone (P < 0. 05) , and copper chelating agent ATTM could reverse the inhibitory effect of ES + Cu2 + on hepatocellular carcinoma cells (P < 0. 05) . Flow cytometry results showed that compared with the control group , the proportion of cell death in PLC/PRF/5 and Huh_7 cells treated with ES + Cu2 + increased , while the proportion of cell death decreased after ATTM intervention (P < 0. 05) . The results of cell scratch test showed that the migration ability of PLC/PRF/5 and Huh_7 cells was decreased after ES + Cu2 + treatment , however, the addition of ATTM reversed the inhibitory effect of ES + Cu2 + on cell migration (P < 0. 05) . Compared with the control group , the expression levels of copper death related proteins ATP7B , FDX1 , DLAT and SOD1 decreased after ES + Cu2 + treatment , but the addition of ATTM reversed the expression trend of these proteins (P < 0. 05) .
Conclusion
The combination of ES and Cu2 + can effectively inhibit the proliferation and migration of PLC/PRF/5 and Huh_7 of hepatocellular carcinoma cells , and induce Cuproptosis , which provides a new strategy for the treatment of hepatocellular carcinoma.
4.Comparison of Clinical and Radiographic Outcomes Between Transforaminal Endoscopic Lumbar Discectomy and Microdiscectomy: A Follow-up Exceeding 5 Years
Xinyu YANG ; Shijun ZHANG ; Junxiao SU ; Sai GUO ; Yakubu IBRAHIM ; Kai ZHANG ; Yonghao TIAN ; Lianlei WANG ; Suomao YUAN ; Xinyu LIU
Neurospine 2024;21(1):303-313
Objective:
To compare the long-term clinical and radiographic outcomes of transforaminal endoscopic lumbar discectomy (TELD) versus microdiscectomy (MD).
Methods:
The data of 154 patients with lumbar disc herniation (LDH) who underwent TELD (n = 89) or MD (n = 65) were retrospectively analyzed. The patients’ clinical outcomes were evaluated using visual analogue scales for leg and low back pain, the Japanese Orthopaedic Association (JOA) score, and the Oswestry Disability Index (ODI). The evolution of radiographic manifestations was observed during follow-up. Potential risk factors for a poor clinical outcome were investigated.
Results:
During a mean follow-up of 5.5 years (range, 5–7 years), the recurrence rate was 4.49% in the TELD group and 1.54% in the MD group. All scores significantly improved from preoperatively to postoperatively in both groups (p < 0.01). The improvement in the ODI and JOA scores was significantly greater in the TELD than MD group (p < 0.05). Forty-seven patients (52.8%) in the TELD group and 32 (49.2%) in the MD group had Modic changes before surgery, most of which showed no changes at the last follow-up. The degeneration grades of 292 discs (71.0%) were unchanged at the last follow-up, while 86 (20.9%) showed improvement, mostly at the upper adjacent segment. No significant difference was observed in the intervertebral height index or paraspinal muscle-disc ratio.
Conclusion
Both TELD and MD provide generally satisfactory long-term clinical outcomes for patients with LDH. TELD can be used as a reliable alternative to MD with less surgical trauma. Modic type II changes, decreased preoperative intervertebral height, and a high body mass index are predictors of a poor prognosis.
5.Comparison of Clinical and Radiographic Outcomes Between Transforaminal Endoscopic Lumbar Discectomy and Microdiscectomy: A Follow-up Exceeding 5 Years
Xinyu YANG ; Shijun ZHANG ; Junxiao SU ; Sai GUO ; Yakubu IBRAHIM ; Kai ZHANG ; Yonghao TIAN ; Lianlei WANG ; Suomao YUAN ; Xinyu LIU
Neurospine 2024;21(1):303-313
Objective:
To compare the long-term clinical and radiographic outcomes of transforaminal endoscopic lumbar discectomy (TELD) versus microdiscectomy (MD).
Methods:
The data of 154 patients with lumbar disc herniation (LDH) who underwent TELD (n = 89) or MD (n = 65) were retrospectively analyzed. The patients’ clinical outcomes were evaluated using visual analogue scales for leg and low back pain, the Japanese Orthopaedic Association (JOA) score, and the Oswestry Disability Index (ODI). The evolution of radiographic manifestations was observed during follow-up. Potential risk factors for a poor clinical outcome were investigated.
Results:
During a mean follow-up of 5.5 years (range, 5–7 years), the recurrence rate was 4.49% in the TELD group and 1.54% in the MD group. All scores significantly improved from preoperatively to postoperatively in both groups (p < 0.01). The improvement in the ODI and JOA scores was significantly greater in the TELD than MD group (p < 0.05). Forty-seven patients (52.8%) in the TELD group and 32 (49.2%) in the MD group had Modic changes before surgery, most of which showed no changes at the last follow-up. The degeneration grades of 292 discs (71.0%) were unchanged at the last follow-up, while 86 (20.9%) showed improvement, mostly at the upper adjacent segment. No significant difference was observed in the intervertebral height index or paraspinal muscle-disc ratio.
Conclusion
Both TELD and MD provide generally satisfactory long-term clinical outcomes for patients with LDH. TELD can be used as a reliable alternative to MD with less surgical trauma. Modic type II changes, decreased preoperative intervertebral height, and a high body mass index are predictors of a poor prognosis.
6.Application of dual-model strategy in image intelligent diagnosis of nail diseases
Junxiao CHEN ; Jie YIN ; Dongying HU ; Zhao WU ; Xiuyan ZHU ; Shiyong WANG
Academic Journal of Naval Medical University 2024;45(8):981-989
Objective To explore a method to improve the accuracy and generalization ability of medical diagnostic neural network models under conditions of small data volumes,and to address the issue of poor neural network model performance in computer-aided diagnosis of nail diseases due to limited training data.Methods A dual-model strategy integrating instance segmentation with fine-grained feature classification was proposed.The neural network model based on dual-model strategy was trained using the dataset of Image-Based Intelligent Diagnosis of Nail Disease Model task of the first National Digital Health Innovation Application Competition & Health and Medical Big Data Theme Competition.This dataset covered 8 types of nail diseases,including nail matrix nevi,paronychia,nail psoriasis,onychomycosis,subungual hemorrhage,melanonychia,periungual warts,and nail melanoma,with class imbalance present.The diagnostic performance of the dual-model strategy was evaluated and compared with single-model strategies(image classification models[ResNet50 and Swin Transformer]and target detection model based on faster region-based convolutional neural network[Faster R-CNN])under the same hardware and software training conditions.Results The dataset included 1 048 samples,including 210 cases of nail matrix nevi,186 cases of paronychia,69 cases of nail psoriasis,203 cases of onychomycosis,149 cases of subungual hemorrhage,71 cases of melanonychia,93 cases of periungual warts,and 67 cases of nail melanoma,with 90%used for training various models and 10%for evaluation.The micro F1 score was 0.324 in the image classification model based on ResNet50,0.381 in the image classification model based on Swin Transformer,0.572 in the target detection model based on Faster R-CNN,and 0.714 in the dual-model strategy model Mask R-CNN+Swin Transformer.The accuracy rates for diagnosing different nail diseases in the dual-model strategy were:nail matrix nevi 80.95%(17/21),paronychia 89.47%(17/19),nail psoriasis 100.00%(7/7),onychomycosis 70.00%(14/20),subungual hemorrhage 73.33%(11/15),melanonychia 14.29%(1/7),periungual warts 55.56%(5/9),and nail melanoma 42.86%(3/7).The micro F1 score for evaluating the dual-model strategy on a test set of 1 000 cases was 0.844.Conclusion The dual-model strategy can effectively combine models with different functions to well accomplish the task of intelligent diagnosis of nail diseases under small data volume training conditions.
7.Comparison of Clinical and Radiographic Outcomes Between Transforaminal Endoscopic Lumbar Discectomy and Microdiscectomy: A Follow-up Exceeding 5 Years
Xinyu YANG ; Shijun ZHANG ; Junxiao SU ; Sai GUO ; Yakubu IBRAHIM ; Kai ZHANG ; Yonghao TIAN ; Lianlei WANG ; Suomao YUAN ; Xinyu LIU
Neurospine 2024;21(1):303-313
Objective:
To compare the long-term clinical and radiographic outcomes of transforaminal endoscopic lumbar discectomy (TELD) versus microdiscectomy (MD).
Methods:
The data of 154 patients with lumbar disc herniation (LDH) who underwent TELD (n = 89) or MD (n = 65) were retrospectively analyzed. The patients’ clinical outcomes were evaluated using visual analogue scales for leg and low back pain, the Japanese Orthopaedic Association (JOA) score, and the Oswestry Disability Index (ODI). The evolution of radiographic manifestations was observed during follow-up. Potential risk factors for a poor clinical outcome were investigated.
Results:
During a mean follow-up of 5.5 years (range, 5–7 years), the recurrence rate was 4.49% in the TELD group and 1.54% in the MD group. All scores significantly improved from preoperatively to postoperatively in both groups (p < 0.01). The improvement in the ODI and JOA scores was significantly greater in the TELD than MD group (p < 0.05). Forty-seven patients (52.8%) in the TELD group and 32 (49.2%) in the MD group had Modic changes before surgery, most of which showed no changes at the last follow-up. The degeneration grades of 292 discs (71.0%) were unchanged at the last follow-up, while 86 (20.9%) showed improvement, mostly at the upper adjacent segment. No significant difference was observed in the intervertebral height index or paraspinal muscle-disc ratio.
Conclusion
Both TELD and MD provide generally satisfactory long-term clinical outcomes for patients with LDH. TELD can be used as a reliable alternative to MD with less surgical trauma. Modic type II changes, decreased preoperative intervertebral height, and a high body mass index are predictors of a poor prognosis.
8.Comparison of Clinical and Radiographic Outcomes Between Transforaminal Endoscopic Lumbar Discectomy and Microdiscectomy: A Follow-up Exceeding 5 Years
Xinyu YANG ; Shijun ZHANG ; Junxiao SU ; Sai GUO ; Yakubu IBRAHIM ; Kai ZHANG ; Yonghao TIAN ; Lianlei WANG ; Suomao YUAN ; Xinyu LIU
Neurospine 2024;21(1):303-313
Objective:
To compare the long-term clinical and radiographic outcomes of transforaminal endoscopic lumbar discectomy (TELD) versus microdiscectomy (MD).
Methods:
The data of 154 patients with lumbar disc herniation (LDH) who underwent TELD (n = 89) or MD (n = 65) were retrospectively analyzed. The patients’ clinical outcomes were evaluated using visual analogue scales for leg and low back pain, the Japanese Orthopaedic Association (JOA) score, and the Oswestry Disability Index (ODI). The evolution of radiographic manifestations was observed during follow-up. Potential risk factors for a poor clinical outcome were investigated.
Results:
During a mean follow-up of 5.5 years (range, 5–7 years), the recurrence rate was 4.49% in the TELD group and 1.54% in the MD group. All scores significantly improved from preoperatively to postoperatively in both groups (p < 0.01). The improvement in the ODI and JOA scores was significantly greater in the TELD than MD group (p < 0.05). Forty-seven patients (52.8%) in the TELD group and 32 (49.2%) in the MD group had Modic changes before surgery, most of which showed no changes at the last follow-up. The degeneration grades of 292 discs (71.0%) were unchanged at the last follow-up, while 86 (20.9%) showed improvement, mostly at the upper adjacent segment. No significant difference was observed in the intervertebral height index or paraspinal muscle-disc ratio.
Conclusion
Both TELD and MD provide generally satisfactory long-term clinical outcomes for patients with LDH. TELD can be used as a reliable alternative to MD with less surgical trauma. Modic type II changes, decreased preoperative intervertebral height, and a high body mass index are predictors of a poor prognosis.
9.Comparison of Clinical and Radiographic Outcomes Between Transforaminal Endoscopic Lumbar Discectomy and Microdiscectomy: A Follow-up Exceeding 5 Years
Xinyu YANG ; Shijun ZHANG ; Junxiao SU ; Sai GUO ; Yakubu IBRAHIM ; Kai ZHANG ; Yonghao TIAN ; Lianlei WANG ; Suomao YUAN ; Xinyu LIU
Neurospine 2024;21(1):303-313
Objective:
To compare the long-term clinical and radiographic outcomes of transforaminal endoscopic lumbar discectomy (TELD) versus microdiscectomy (MD).
Methods:
The data of 154 patients with lumbar disc herniation (LDH) who underwent TELD (n = 89) or MD (n = 65) were retrospectively analyzed. The patients’ clinical outcomes were evaluated using visual analogue scales for leg and low back pain, the Japanese Orthopaedic Association (JOA) score, and the Oswestry Disability Index (ODI). The evolution of radiographic manifestations was observed during follow-up. Potential risk factors for a poor clinical outcome were investigated.
Results:
During a mean follow-up of 5.5 years (range, 5–7 years), the recurrence rate was 4.49% in the TELD group and 1.54% in the MD group. All scores significantly improved from preoperatively to postoperatively in both groups (p < 0.01). The improvement in the ODI and JOA scores was significantly greater in the TELD than MD group (p < 0.05). Forty-seven patients (52.8%) in the TELD group and 32 (49.2%) in the MD group had Modic changes before surgery, most of which showed no changes at the last follow-up. The degeneration grades of 292 discs (71.0%) were unchanged at the last follow-up, while 86 (20.9%) showed improvement, mostly at the upper adjacent segment. No significant difference was observed in the intervertebral height index or paraspinal muscle-disc ratio.
Conclusion
Both TELD and MD provide generally satisfactory long-term clinical outcomes for patients with LDH. TELD can be used as a reliable alternative to MD with less surgical trauma. Modic type II changes, decreased preoperative intervertebral height, and a high body mass index are predictors of a poor prognosis.
10.HIV-1 subtypes and drug-resistance mutation sites in HIV/AIDS patients with antiretroviral-therapy failure
Junxiao LIN ; Hongzhu WANG ; Congcong LI ; Ying SHENG ; Guixia LI
Shanghai Journal of Preventive Medicine 2023;35(3):224-228
ObjectiveTo analyze the characteristics of HIV-1 subtypes and drug-resistance mutation sites among HIV-infected patients who received high-efficiency antiretroviral therapy but failed. MethodsA total of 130 plasma samples were collected from the patients who received antiviral treatment for 6 months in Taizhou City of Zhejiang Province in 2021 but failed the treatment and the viral load was ≥1 000 copies·mL-1. Nucleic acid in the samples was extracted, and the pol gene was amplified by nested reverse transcription PCR. After next-generation sequencing, online tools were used to compare and analyze the subtypes and drug-resistant mutation sites. ResultsA total of 110 samples were successfully sequenced. The main HIV-1 subtype was CRF01_AE, accounting for 42.72% (47 cases), followed by CRF07_BC, 35.45% (39 cases); CRF08_BC, 10.00% (11 cases); CRF85_BC , 8.18% (9); and a small number of B subtype, 1.81% (2 cases) and C subtype, 1.81% (2 cases). The online tool comparison showed that there were 67 cases with mutations of drug-resistance sites and 61 cases with drug-resistance. The mutation sites were mainly M184V, K103N, K65R and V181C, and the mutation rates were 20.00% (22 cases), 10.91% (12 cases), 8.18% (9 cases) and 8.18% (9 cases), respectively. These mutation sites caused different degrees of resistance to nucleoside reverse transcriptase inhibitors (NRTI), non- nucleoside reverse transcriptase inhibitors (NNRTI) and protease inhibitors (PI), including 45 cases of NRTI, 61 cases of NNRTI and 2 cases of PI resistance. ConclusionThe HIV infected people who fail the treatment in Taizhou are mainly with the subtypes CRF01_AE and CRF07_BC. The rate of drug-resistance mutation is at a moderate level, mainly due to the mutation of NRTI and NNRTI drug-resistance sites, and a small number of PI drug-resistance sites. Therefore, the antiviral treatment plan for HIV infected people should be reasonably adjusted, and the detection of drug-resistance mutation sites should be strengthened to avoid the generation of transmissible drug-resistance strains.


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