1.Lycorine promotes ferroptosis in osteosarcoma cells via YAP1/TRFC path-way and enhances sensitivity to cisplatin
Linkuan WANG ; Xinping WANG ; Kaijun YI ; Zhiyong LI ; Qi ZHAO ; Yi-hua SHI
Chinese Journal of Pathophysiology 2025;41(5):882-894
AIM:This study aims to investigate the sensitizing effects of lycorine(Lyc)in combination with cisplatin(Cis)on human osteosarcoma cells and to explore the underlying mechanisms of action.METHODS:Human osteosarcoma cell lines MG63,HOS,and cisplatin-resistant HOS/DDP cells were utilized to evaluate the effects of Lyc and cisplatin,both alone and in combination,on cell viability using the CCK8 assay.The clonogenic assay was performed to assess cell proliferation capacity,while the scratch assay evaluated the drugs' effects on cell migration.Reactive oxygen species(ROS)levels were measured using a ROS assay kit,and changes in intracellular glutathione(GSH)levels were assessed with a GSH/oxidized glutathione(GSSG)assay kit.The mitochondrial membrane potential was analyzed via JC-1 staining to determine the drugs' effects on mitochondrial function.Intracellular iron(Ⅱ)content changes were detected us-ing FerrOrange,a fluorescence probe,and cellular malondialdehyde(MDA)levels were measured using an MDA assay kit.RT-qPCR was employed to evaluate the expression levels of key genes related to ferroptosis,and Western blot analysis was conducted to detect changes in the protein expression levels of Yes-associated protein 1(YAP1),acyl-CoA synthetase long-chain family member 4(ACSL4),glutathione peroxidase 4(GPX4),heme oxygenase-1(HO-1),and transferrin re-ceptor(TFRC).RESULTS:Both Lyc and cisplatin effectively inhibited the proliferation of human osteosarcoma cells.Notably,the combination of Lyc and cisplatin led to a more substantial reduction in cell viability,proliferation,and migra-tion abilities in MG63,HOS,and HOS/DDP cells compared to cisplatin alone.Additionally,this combination significant-ly increased ROS levels while decreasing GSH content,indicating mitochondrial damage and elevated iron(Ⅱ)and MDA levels.RT-qPCR results revealed that the combination treatment more significantly downregulated ferroptosis-promoting genes and upregulated ferroptosis-inhibiting genes compared to cisplatin treatment alone(P<0.05).Western blot results showed a slight decrease in GPX4 protein expression following Lyc and cisplatin treatment,while expression levels of YAP1,TFRC,ACSL4,and HO-1 were significantly increased(P<0.05).CONCLUSION:Lyc enhances the sensitivi-ty of MG63,HOS,and HOS/DDP cells to cisplatin by promoting ferroptosis through the YAP1/TFRC signaling pathway.
2.Lycorine promotes ferroptosis in osteosarcoma cells via YAP1/TRFC path-way and enhances sensitivity to cisplatin
Linkuan WANG ; Xinping WANG ; Kaijun YI ; Zhiyong LI ; Qi ZHAO ; Yi-hua SHI
Chinese Journal of Pathophysiology 2025;41(5):882-894
AIM:This study aims to investigate the sensitizing effects of lycorine(Lyc)in combination with cisplatin(Cis)on human osteosarcoma cells and to explore the underlying mechanisms of action.METHODS:Human osteosarcoma cell lines MG63,HOS,and cisplatin-resistant HOS/DDP cells were utilized to evaluate the effects of Lyc and cisplatin,both alone and in combination,on cell viability using the CCK8 assay.The clonogenic assay was performed to assess cell proliferation capacity,while the scratch assay evaluated the drugs' effects on cell migration.Reactive oxygen species(ROS)levels were measured using a ROS assay kit,and changes in intracellular glutathione(GSH)levels were assessed with a GSH/oxidized glutathione(GSSG)assay kit.The mitochondrial membrane potential was analyzed via JC-1 staining to determine the drugs' effects on mitochondrial function.Intracellular iron(Ⅱ)content changes were detected us-ing FerrOrange,a fluorescence probe,and cellular malondialdehyde(MDA)levels were measured using an MDA assay kit.RT-qPCR was employed to evaluate the expression levels of key genes related to ferroptosis,and Western blot analysis was conducted to detect changes in the protein expression levels of Yes-associated protein 1(YAP1),acyl-CoA synthetase long-chain family member 4(ACSL4),glutathione peroxidase 4(GPX4),heme oxygenase-1(HO-1),and transferrin re-ceptor(TFRC).RESULTS:Both Lyc and cisplatin effectively inhibited the proliferation of human osteosarcoma cells.Notably,the combination of Lyc and cisplatin led to a more substantial reduction in cell viability,proliferation,and migra-tion abilities in MG63,HOS,and HOS/DDP cells compared to cisplatin alone.Additionally,this combination significant-ly increased ROS levels while decreasing GSH content,indicating mitochondrial damage and elevated iron(Ⅱ)and MDA levels.RT-qPCR results revealed that the combination treatment more significantly downregulated ferroptosis-promoting genes and upregulated ferroptosis-inhibiting genes compared to cisplatin treatment alone(P<0.05).Western blot results showed a slight decrease in GPX4 protein expression following Lyc and cisplatin treatment,while expression levels of YAP1,TFRC,ACSL4,and HO-1 were significantly increased(P<0.05).CONCLUSION:Lyc enhances the sensitivi-ty of MG63,HOS,and HOS/DDP cells to cisplatin by promoting ferroptosis through the YAP1/TFRC signaling pathway.
3.Tubridge flow diverter for the treatment of recurrent internal carotid blood blister-like aneurysms after stent-assisted embolization
Yina WU ; Zhiwen LU ; Guoli DUAN ; Yibin FANG ; Kaijun ZHAO ; Yi XU ; Qinghai HUANG
International Journal of Cerebrovascular Diseases 2021;29(10):750-754
Objective:To investigate the safety and efficacy of Tubridge flow diverter for the treatment of recurrent internal carotid blood blister-like aneurysms after stent-assisted embolization.Methods:From June 2018 to April 2021, patients with recurrent internal carotid blood blister-like aneurysms treated with Tubridge flow diverter in the Department of Neurosurgery, Changhai Hospital, Naval Medical University were enrolled retrospectively. The perioperative safety, immediate postoperative and follow-up results were analyzed.Results:A total of 6 patients with recurrent internal carotid blood blister-like aneurysm after stent-assisted embolization were enrolled. The time interval from the first stent-assisted embolization to Tubridge placement was 14 to 90 d. Tubridge implantation alone was used in 4 patients, and Tubridge was implanted in the other 2 patients after the coils were packed. There were no complications during the perioperative period, and no rebleeding was observed after clinical follow-up for 5 to 36 months. Five patients were followed up by angiography for 1-3 months, and the aneurysms disappeared completely.Conclusion:Tubridge flow diverter for the treatment of recurrent internal carotid blood blister-like aneurysms is safe and effective.
4.Early management of complication on severe multiple trauma
Fan YANG ; Xiangjun BAI ; Kaijun HU ; Zhaohui TANG ; Chengla YI ; Yiliu LIAO
Chinese Journal of Emergency Medicine 2009;18(6):628-631
Objective To analyze the efficacy of early management of eomplications in patients with multi-ple traumas by applying the damage control theory. Method All total of 116 patients admitted to Wuhan Tongji Hospital between Jan 2006 and Jan 2008 were included in this retrospective study and were classified according to treatment method. Overall, 55 cases underwent damage control operations and 61 cases underwent conventional management. The groups of patients were compared in terms of parameters such as length of operation time, time to resuscitate from shock, lethal triad of death (LTD), complications and treatment outcomes after operation. Results The length of operation time was (67.43±19.52) min, resuscitation time from shock was (6.77±3.16) h and LTD was (11.54±4.10) h in the damage control group, and (163.95±55.41) min, (22.51±11.65) h and (34.55±5.63) h, respectively, in the conventional group, with significant differences between the two groups (P< 0.01). The per capita complication and mortality rates were 227.27 % and 3.64 %, respectively, in the dam-age control group and 363.93 % and 9.84%, respectively, in the conventional group (P <0.05). Couclusions Here, we demonstrated that early management of patients with multiple traumas undergoing operations by apply-ing the damage control theory significantly decreased the operation time, and decreased time to resuscitation from shock and LTD. Furthermore, this approach significantly reduced the incidence of comphcations and the mortality rate of patients with severe multiple traumas.
5.Effect of thymosin ?1 on percentage of T-lymphocyte subsets in in vitro peripheral blood mononuclear cells from gastric cancer patients
Xia YANG ; Feng QIAN ; Kaijun LIU ; Haiyang HE ; Yuanzhi LAN ; Yi TIAN ; Xiaolan FU ; Jian LI ; Ji ZHANG ; Zigang SHEN ; Jintao LI ; Yuzhang WU
Journal of Third Military Medical University 2002;0(12):-
Objective To investigate the effect of thymosin ?1 (T?1) on cellular immune function in gastric cancer patients through observing its treatment on the differentiation of T-lymphocyte subsets from screened peripheral blood mononuclear cells (PBMCs). Methods PBMCs were obtained by centrifugation of blood samples from 18 healthy subjects and 32 patients with gastric cancer,and then cultured in the presence of culture medium with addition of T?1 at 50,10 and 1 ?g/ml for 2 d. T lymphocyte subsets (such as CD4+,CD8+ and CD4+ CD25+ Foxp3+ T cells) and Th1/Th2 multiplex cytokines were detected by flow cytometry (FCM). Results After PBMCs isolated from healthy people and patients were incubated with or without T?1,there was no significant change in percentage of CD4+,CD8+ peripheral lymphocyte subsets and ratio of CD4+/CD8+. There was no obvious change in the percentage of CD4+ CD25+ Foxp3+ T lymphocyte subsets in the normal control,but a significant increase was observed in the cells from patients with gastric cancer after treatment (P

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