Ferroptosis, a new form of programmed cell death marked by iron-dependent phospholipid peroxidation, is regulated by complex cellular metabolic pathways, including iron metabolism, lipid metabolism, and oxidation-reduction system, is associated with many organ injuries and degeneration, and has great potential in the treatment of ischemic diseases and lipid peroxide-related degenerative diseases. Myocardial ischemia reperfusion injury (MIRI) is the most common cause of death in patients with acute myocardial infarction after revascularization therapy. Recent studies have shown that ferroptosis is intimately related to the pathological process of MIRI. Ferroptosis is associated with MIRI through oxidative stress, iron metabolism, lipid metabolism, endoplasmic reticulum stress and inflammatory response. Intervention of ferroptosis during reperfusion can effectively improve cardiac function and reduce myocardial infarct size. In this paper, the research progress was explored between ferroptosis and MIRI, and the specific role of ferroptosis in MIRI was discussed.

Objective:To explore the clinical efficacy and risk factors of umbilical cord mesenchymal stem cells (UCMSCs) infusion at an early stage (i.e.gross hematuria) for hemorrhagic cystitis (HC) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:The relevant clinical data were retrospectively reviewed for 300 patients undergoing allo-HSCT from January 2016 to July 2021.According to the presence or absence of HC, they were assigned into two groups of HC (n=89) and non-HC (control, n=211). According to whether or not receiving an infusion of UCMSCs, 51 patients of HC degree Ⅱ-Ⅳ were divided into two groups of UCMSC infusion and non-infusion.The risk factors of HC after allo-HSCT were analyzed by χ2 test.Logistic regression was employed for multivariate analysis of P<0.05.Mann-Whitney U test was utilized for statistically analyzing the duration of gross hematuria and urinary tract irritation symptoms and evaluating the clinical efficacy of UCMSCs infusion for HC. Results:Among them, 89 (29.67%) developed HC post-allo-HSCT.Clinical grades were Ⅰ (n=38, 42.70%), Ⅱ (n=36, 40.45%), Ⅲ (n=13, 14.61%) and Ⅳ (n=2, 2.25%). The median occurrence time was 29 (21.5-35.0) days post-allo-HSCT.In univariate analysis, age ≤30 years, haploid transplantation, antithymocyte globulin (ATG), acute graft-versus-host disease (aGVHD), CMV-DNA positive pretreatment significantly boosted the risk of HC ( P<0.05). In multivariate analysis, aGVHD was an independent risk factor for HC ( OR=10.281, 95% CI: 1.606-65.813, P=0.014). Among 89 HC patients, 38 grade Ⅰ patients were complete remission(CR). Among 51 patients of grade Ⅱ-Ⅳ HC, the outcomes were CR (n=48) and non-remission(NR)(n=3). And 24/51 of them received UCMSCs plus conventional treatment.The duration of gross hematuria was shorter in UCMSCs infusion group than that in UCMSCs non-infusion group [12(9-17) vs 17(12.0-26.5) day] and the difference was statistically significant ( P=0.045). And the duration of urinary tract irritation symptoms was shorter in UCMSCs infusion group than that in UCMSCs non-infusion group [18(11-30) vs 27(18.0-35.5) days] and the difference was statistically significant ( P=0.048). Conclusions:Indicated for post-ALLO-HSCT HC, infusion of UCMSCs may significantly shorten the course of disease.Age ≤30 years, haploid transplantation and preconditioning with positive ATG, aGVHD and CMV-DNA may boost the risks of HC post-allo-HSCT.And aGVHD is an independent risk factor for HC after allo-HSCT.

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[Abstract] Objective: To develop a new type of CD7 chimeric antigen receptor modified T cell (CD7-CAR-T) for the treatment of CD7 positive acute myeloid leukemia (AML), and to observe its killing effect on CD7 positive AML cells. Methods: The CD7-CAR lentiviral vector was constructed based on the CD7 Nanobody sequence and costimulatory domain sequence of CD28 and 4-1BB. The lentiviral particles were packaged and used to co-transfect human T cells with protein expression blocker (PEBL), so as to prepare CD7- CAR-T cells. Real time cellular analysis (RTCA) was used to monitor the cytotoxicity of CD7-CAR-T cells on CD7 overexpressed 293T cells. Flow cytometry assay was used to detect the effect of CD7-CAR-T cells on proliferation and cytokine secretion of AML cells with high, medium and low CD7 expressions (KG-1, HEL and Kasumi-1 cells, respectively). Results: CD7-CAR-T cell was successfully constructed and its surface expression of CD7 was successfully blocked. Compared with T cells, CD7-CAR-T cells could significantly inhibit the proliferation of CD7-293T cells and promote the release of TNF, Granzyme B and INF-γ; in addition, CD7-CAR-T cells also significantly promoted the apoptosis (t=147.1, P<0.01; t=23.57, P<0.01) and cytokine release (P<0.05 or P<0.01) in CD7 positive KG-1 and HEL cells, but had little effect on Kasumi-1 cells that only expressed minimal CD7 antigen (t=0.7058, P>0.05). Conclusion: CD7-CAR-T cells can specifically kill CD7-positive AML cells in vitro.

Objective:To explore the mechanism of inflammatory response in lung tissues of rats with decompression sickness (DCS) caused by fast buoyancy ascent escape, and to observe the changes in phosphorylation of extracellular signal-related kinase (p-ERK) expression in rats with early onset of DCS during a simulated fast buoyancy ascent escape.Methods:A total of 18 rats were divided into three groups randomly, i. e. control group (C group), escape group in which all six rats were dead (ED group), and escape group in which all six rats survived (ES group). The rats in the ES group were anesthetized and sacrificed 8 minutes after they managed the simulated insecure seawater fast buoyancy ascent escape in a hyperbaric chamber. The p-ERK protein was extracted from rat lungs and then the protein expression was analyzed by Western blotting.Results:The p-ERK expression level in the ED group was elevated significantly than that in the C group; while the p-ERK expression level in ES group was not significantly increased than that in the C group.Conclusion:The development of inflammation-like reaction in lung tissues of rats with DCS caused by fast buoyancy ascent escape may relate with the activation of p-ERK.

Objective:To explore the mechanism of inflammatory response in lung tissues of rats with decompression sickness (DCS) caused by fast buoyancy ascent escape, and to observe the changes in phosphorylation of extracellular signal-related kinase (p-ERK) expression in rats with early onset of DCS during a simulated fast buoyancy ascent escape.Methods:A total of 18 rats were divided into three groups randomly, i. e. control group (C group), escape group in which all six rats were dead (ED group), and escape group in which all six rats survived (ES group). The rats in the ES group were anesthetized and sacrificed 8 minutes after they managed the simulated insecure seawater fast buoyancy ascent escape in a hyperbaric chamber. The p-ERK protein was extracted from rat lungs and then the protein expression was analyzed by Western blotting.Results:The p-ERK expression level in the ED group was elevated significantly than that in the C group; while the p-ERK expression level in ES group was not significantly increased than that in the C group.Conclusion:The development of inflammation-like reaction in lung tissues of rats with DCS caused by fast buoyancy ascent escape may relate with the activation of p-ERK.

Objective:To prepare nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR and to examine its cytotoxicity against EGFR positive tumor cells.Methods:By using molecular cloning strategy,prokaryotic expression construct of pET28a-BI7D12-PE38KDEL was generated which consisted of nanobody 7D12 targeting EGFR in the form of a divalent fused with PE38KDEL,a truncated form of pseudomonas exotoxin A via a flexible peptide(G4S)4,and then transformed into E.coli BL21(DE3).Protein expression was induced by adding IPTG,purified by Ni-affinity column chromatography,and verified by Western blot.The binding capacity of the resulted immunotoxin to EGFR-positive cells A549,HT29,MCF-7 and EGFR-negative cells CEM,Jurkat were determined by flow cytometry assay,and its cytotoxicity against the target cells was examined.Briefly,tumor cells were treated with different dosage of the immunotoxin,and the killing efficacy of BI7D12-PE38KDEL on these cells were assessed by WST-1 assay after 72 hours.Results:The SDS-PAGE and Western blot results showed the recombinant immunotoxin BI7D12-PE38KDEL was successfully prepared,and majority of them was expressed in soluble form.BI7D12-PE38KDEL could selectively bind to EGFR-positive cells of A549,HT29,and MCF-7.More importantly,the immunotoxin exhibited much more significant killing effect on these EGFR positive cells compared to the negative control group of CEM and Jurkat cells(P<0.01).Conclusion:In the current study,the nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR was successfully prepared and exhibited a superior inhibition effect for the growth of EGFR-positive cells.

Objective To study the therapeutic effect of hyperbaric oxygen HBO) combined with repetitive transcranial magnetic stimulation (rTMS) in the treatment of coma following craniocerebral trauma.Methods One hundred and twenty coma patients caused by craniocerebral trauma were randomly selected for the study.Those patients without treatment or without rTMS were assigned as the control groups,and the group with Glasgow coma scale (GCS) scures of 6-8 was further assigned as the CA group,and the group with GCS scores of 3-5 was assigned as the CB group.In those patients who received HBO therapy,the ones with GCS scores of 6-8 were assigned as the HA group,and the ones with GCS scores of 3-5 were assigned as the HB group (conventional therapy group B).In those patients who received both HBO and rTMS therapy,the ones with GCS scores of 6-8 were assigned as the rTA group,while the ones with GCS scores of 3-5 were assigned as the rTB group.The patients of the 6 groups were given conventional therapy for the treatment of brain trauma,such as neural nutrition,anti-infection and maintenance of electrodes.The patients in the HA group and HB group were additionally treated with HBO,while the patients in the rTA and rTB groups were further treated with HBO and rTMS.The patients received HBO therapy at a pressure of 0.25 MPa,once a day,10 days a treatment course,for a succession of 3 courses.The stimulation intensity of the rTMS treatment protocol was set at 0.72 T,with the frequency of the magnetic field being 0.5 HZ.The patients had 30 stimulations for every sequence,one sequence a day,12 days a treatment course for a succession of 3 courses.Scores of Glasgow Coma Scale (GCS) and Glasgow Outcome Scale (GOS) before and after treatment were assessed and statistically analyzed.Results After 3 successive treatment courses,the patients with GCS scores of 6-8 were further treated with HBO combined with rTMS in addition to conventional therapy.The GCS scores of the HBO + rTMS + conventional therapy group were as high as(11.15 ± 4.17),which were higher than the scores of the HBO + conventional therapy group (9.05 ± 3.12),which were also higher than the scores of the conventional therapy group (7.50 ± 1.99).Statistical significance could be noted,when comparisons were made between the 3 groups(P ＜ 0.05).GOS scores of the HBO + rTMS + conventional therapy group (4.45 ± 0.83) were higher than those of the conventional therapy group (3.90 ± 0.55),with statistical significance (P ＜0.05).Conclusions The patients with GCS scores of 6-8 treated with HBO + rTMS + conventional therapy had better rehabilitation effect and prognosis,and therapeutic results were obviously superior to those of the patients treated with simple conventional therapy and HBO + conventional therapy.

Objective To study the therapeutic effect of hyperbaric oxygen HBO) combined with repetitive transcranial magnetic stimulation (rTMS) in the treatment of coma following craniocerebral trauma.Methods One hundred and twenty coma patients caused by craniocerebral trauma were randomly selected for the study.Those patients without treatment or without rTMS were assigned as the control groups,and the group with Glasgow coma scale (GCS) scures of 6-8 was further assigned as the CA group,and the group with GCS scores of 3-5 was assigned as the CB group.In those patients who received HBO therapy,the ones with GCS scores of 6-8 were assigned as the HA group,and the ones with GCS scores of 3-5 were assigned as the HB group (conventional therapy group B).In those patients who received both HBO and rTMS therapy,the ones with GCS scores of 6-8 were assigned as the rTA group,while the ones with GCS scores of 3-5 were assigned as the rTB group.The patients of the 6 groups were given conventional therapy for the treatment of brain trauma,such as neural nutrition,anti-infection and maintenance of electrodes.The patients in the HA group and HB group were additionally treated with HBO,while the patients in the rTA and rTB groups were further treated with HBO and rTMS.The patients received HBO therapy at a pressure of 0.25 MPa,once a day,10 days a treatment course,for a succession of 3 courses.The stimulation intensity of the rTMS treatment protocol was set at 0.72 T,with the frequency of the magnetic field being 0.5 HZ.The patients had 30 stimulations for every sequence,one sequence a day,12 days a treatment course for a succession of 3 courses.Scores of Glasgow Coma Scale (GCS) and Glasgow Outcome Scale (GOS) before and after treatment were assessed and statistically analyzed.Results After 3 successive treatment courses,the patients with GCS scores of 6-8 were further treated with HBO combined with rTMS in addition to conventional therapy.The GCS scores of the HBO + rTMS + conventional therapy group were as high as(11.15 ± 4.17),which were higher than the scores of the HBO + conventional therapy group (9.05 ± 3.12),which were also higher than the scores of the conventional therapy group (7.50 ± 1.99).Statistical significance could be noted,when comparisons were made between the 3 groups(P ＜ 0.05).GOS scores of the HBO + rTMS + conventional therapy group (4.45 ± 0.83) were higher than those of the conventional therapy group (3.90 ± 0.55),with statistical significance (P ＜0.05).Conclusions The patients with GCS scores of 6-8 treated with HBO + rTMS + conventional therapy had better rehabilitation effect and prognosis,and therapeutic results were obviously superior to those of the patients treated with simple conventional therapy and HBO + conventional therapy.