AIM:To investigate the effect of intrinsic specific transforming growth factor-β(TGF-β)signaling on regeneration and repair of myofibers in acute skeletal myositismice model induced by cardiotoxin(CTX).METHODS:One hundred and eighty-six wild C57BL/6 mice and one hundred and thirty-eight mice with conditional knockout of TGF-β receptor 2(TGF-βr2)in myofibers(SM TGF-βr2-/-mice)were selected.CTX injection to anterior tibial muscle(TA)in-duced acute myoinjury in mice.Some SM TGF-βr2-/-mice were given Smad signaling agonist SRI-011381(SRI)intramus-cular injection.All mice were mainly divided into the following groups:control group,SM TGF-βr2-/-group and SM TGF-βr2-/-+SRI group.Twenty-four mice were selected in each group.RT-qPCR and immunofluorescence staining were used to detect the relative mRNA level,protein expression of inflammatory cytokines and low-density lipoprotein receptor-related protein 1(LRP1),respectively,while the relative protein expression of myosin heavy chain 3(MHY3)and embryonic myosine heavy chain(eMHC)in damaged muscle was detected by Western blot and immunofluorescence staining.In vi-tro,after being extracted from neonatal mice,myogenic precursor cells(MPCs)were cultured in an pro-inflammatory mi-lieu and treated with SRI,recombinant mouse extracellular matrix protein 1(rmECM1)alone or in combination.Hereby,they were divided into the following seven groups:control-MPCs group,control-MPCs+LPS group,TGF-βr2-/--MPCs group,TGF-βr2-/--MPCs+LPS group,TGF-βr2-/--MPCs+LPS+SRI group,TGF-βr2-/--MPCs+LPS+rmECM1 group,and TGF-βr2-/--MPCs+LPS+SRI+rmECM1 group.Six mice were selected in each group.RT-qPCR and Western blot were used to detect the relative mRNA level,protein expression of major histocompatibility complex class I molecules(MHC-I/H-2Kb),major histocompatibility complex class II molecules(MHC-II/H2-Eα),Toll-like receptor 3(TLR3),and LRP1.And the relative protein expression of MoyD and myogenin in myotubes was detected by immunofluorescence staining.RE-SULTS:In vivo,compared with control group,SM TGF-βr2-/-group showed the significant upregulation of pro-inflamma-tory cytokines(P<0.05),and the opposite trend of anti-inflammatory cytokines,LRP1,MHY3,eMHC in the injured muscle(P<0.05),with delayed regeneration and repair of myofibers.In vitro,compared with control-MPCs+LPS group,LRP1,MoyD and myogenin significantly downregulated in TGF-βr2-/--MPCs+LPS group,but the downregulation trend was corrected after giving SRI treatment(P<0.05).In addition,compared with the TGF-βr2-/--MPCs+LPS group,the combi-nation of rmECM1 and SRI significantly upregulated the protein expression of MyoD and myogenin(P<0.05).CONCLU-SION:In a mouse model of acute skeletal myositis,intrinsic TGF-β signaling specifically in myofibers regulates local im-mune behavior.It promotes the expression of LRP1 in damaged muscle via Smad2/3 signaling,and LRP1 can then fully bind to ECM1,thereby facilitating muscle regeneration and repair,and improving the prognosis of acute skeletal myositis.

Inflammatory bowel disease(IBD)is a chronic nonspecific inflammatory disease of the intestines,mainly including Crohn's disease(CD)and ulcerative colitis(UC).More than 20％of IBD patients progress to colitis-associated colorectal cancer(CAC)through the inflammation-cancer sequence within 30 years of disease onset.Nuclear transcription factor E2-related factor 2(Nrf2)is a transcription factor closely related to IBD and CAC,which not only affects inflammation but also plays a significant role in regulating the"Inflammation-cancer transformation".In this paper,we review the studies on the mechanism of Nrf2 in the"Inflammation-cancer transformation",focusing on the mechanism and dual roles of Nrf2 in oxidative stress,inflammatory factors,immune response,metabolic reprogramming and gene expression,with an aim of providing a reference for further understanding the potential value of its intervention in the human tumor process.

AIM:To investigate the effect of intrinsic specific transforming growth factor-β(TGF-β)signaling on regeneration and repair of myofibers in acute skeletal myositismice model induced by cardiotoxin(CTX).METHODS:One hundred and eighty-six wild C57BL/6 mice and one hundred and thirty-eight mice with conditional knockout of TGF-β receptor 2(TGF-βr2)in myofibers(SM TGF-βr2-/-mice)were selected.CTX injection to anterior tibial muscle(TA)in-duced acute myoinjury in mice.Some SM TGF-βr2-/-mice were given Smad signaling agonist SRI-011381(SRI)intramus-cular injection.All mice were mainly divided into the following groups:control group,SM TGF-βr2-/-group and SM TGF-βr2-/-+SRI group.Twenty-four mice were selected in each group.RT-qPCR and immunofluorescence staining were used to detect the relative mRNA level,protein expression of inflammatory cytokines and low-density lipoprotein receptor-related protein 1(LRP1),respectively,while the relative protein expression of myosin heavy chain 3(MHY3)and embryonic myosine heavy chain(eMHC)in damaged muscle was detected by Western blot and immunofluorescence staining.In vi-tro,after being extracted from neonatal mice,myogenic precursor cells(MPCs)were cultured in an pro-inflammatory mi-lieu and treated with SRI,recombinant mouse extracellular matrix protein 1(rmECM1)alone or in combination.Hereby,they were divided into the following seven groups:control-MPCs group,control-MPCs+LPS group,TGF-βr2-/--MPCs group,TGF-βr2-/--MPCs+LPS group,TGF-βr2-/--MPCs+LPS+SRI group,TGF-βr2-/--MPCs+LPS+rmECM1 group,and TGF-βr2-/--MPCs+LPS+SRI+rmECM1 group.Six mice were selected in each group.RT-qPCR and Western blot were used to detect the relative mRNA level,protein expression of major histocompatibility complex class I molecules(MHC-I/H-2Kb),major histocompatibility complex class II molecules(MHC-II/H2-Eα),Toll-like receptor 3(TLR3),and LRP1.And the relative protein expression of MoyD and myogenin in myotubes was detected by immunofluorescence staining.RE-SULTS:In vivo,compared with control group,SM TGF-βr2-/-group showed the significant upregulation of pro-inflamma-tory cytokines(P<0.05),and the opposite trend of anti-inflammatory cytokines,LRP1,MHY3,eMHC in the injured muscle(P<0.05),with delayed regeneration and repair of myofibers.In vitro,compared with control-MPCs+LPS group,LRP1,MoyD and myogenin significantly downregulated in TGF-βr2-/--MPCs+LPS group,but the downregulation trend was corrected after giving SRI treatment(P<0.05).In addition,compared with the TGF-βr2-/--MPCs+LPS group,the combi-nation of rmECM1 and SRI significantly upregulated the protein expression of MyoD and myogenin(P<0.05).CONCLU-SION:In a mouse model of acute skeletal myositis,intrinsic TGF-β signaling specifically in myofibers regulates local im-mune behavior.It promotes the expression of LRP1 in damaged muscle via Smad2/3 signaling,and LRP1 can then fully bind to ECM1,thereby facilitating muscle regeneration and repair,and improving the prognosis of acute skeletal myositis.

Objective To investigate the correlation of imaging parameters of CT perfusion scan-ning combined with serum suPAR and Mac-2BP with END and poor outcomes in patients with BAD.Methods A total of 176 BAD patients admitted to Department of Neurology of Zhengzhou People's Hospital from June 2019 to March 2024 were prospectively enrolled,and according to whether END occurred within 72 h after onset,they were divided into END group(42 cases)and non-END group(134 cases).Based on their outcomes at 90 d of follow-up,they were also assigned into good outcome group(129 cases)and poor outcome group(47 cases).The serum suPAR and Mac-2BP levels were compared between the END and non-END groups,as well as the good and poor outcome groups.ROC curves were plotted to analyze the accuracies of serum suPAR and Mac-2BP in predicting END and poor outcomes.Results The END group had significantly higher suPAR and Mac-2BP levels,longer MTT and TTP,and larger proportions of DWMH,basal gan-glia EPVS and cerebral perfusion impairment,but lower CBV and CBF when compared with the non-END group(P＜0.05,P＜0.01).Obviously higher suPAR and Mac-2BP levels,NIHSS score at END,longer MTT and TTP,and larger proportions of EDN,basal ganglia EPVS and cerebral perfusion impairment,but lower CBV and CBF were observed in the poor outcome group than the good outcome group(P＜0.05,P＜0.01).suPAR and Mac-2BP levels,cerebral perfusion impair-ment and DWMH were independent risk factors for END(P＜0.05,P＜0.01).suPAR,Mac-2BP,NIHSS score at END,END,cerebral perfusion impairment,and basal ganglia EPVS were inde-pendent risk factors for poor outcomes(P＜0.05,P＜0.01).The AUC value of combined suPAR and Mac-2BP in predicting END was 0.8844(95％CI:0.822-0.947),and the value in predicting poor outcomes was 0.8742(95％CI:0.810-0.938)in BAD patients.Conclusion Cerebral perfu-sion impairment,suPAR and Mac-2BP are independent risk factors for END and poor outcome at 90 d in BAD patients.Combined detection can is helpful in evaluating the patient's condition and predicting the prognosis.

Objective To investigate the correlation of imaging parameters of CT perfusion scan-ning combined with serum suPAR and Mac-2BP with END and poor outcomes in patients with BAD.Methods A total of 176 BAD patients admitted to Department of Neurology of Zhengzhou People's Hospital from June 2019 to March 2024 were prospectively enrolled,and according to whether END occurred within 72 h after onset,they were divided into END group(42 cases)and non-END group(134 cases).Based on their outcomes at 90 d of follow-up,they were also assigned into good outcome group(129 cases)and poor outcome group(47 cases).The serum suPAR and Mac-2BP levels were compared between the END and non-END groups,as well as the good and poor outcome groups.ROC curves were plotted to analyze the accuracies of serum suPAR and Mac-2BP in predicting END and poor outcomes.Results The END group had significantly higher suPAR and Mac-2BP levels,longer MTT and TTP,and larger proportions of DWMH,basal gan-glia EPVS and cerebral perfusion impairment,but lower CBV and CBF when compared with the non-END group(P＜0.05,P＜0.01).Obviously higher suPAR and Mac-2BP levels,NIHSS score at END,longer MTT and TTP,and larger proportions of EDN,basal ganglia EPVS and cerebral perfusion impairment,but lower CBV and CBF were observed in the poor outcome group than the good outcome group(P＜0.05,P＜0.01).suPAR and Mac-2BP levels,cerebral perfusion impair-ment and DWMH were independent risk factors for END(P＜0.05,P＜0.01).suPAR,Mac-2BP,NIHSS score at END,END,cerebral perfusion impairment,and basal ganglia EPVS were inde-pendent risk factors for poor outcomes(P＜0.05,P＜0.01).The AUC value of combined suPAR and Mac-2BP in predicting END was 0.8844(95％CI:0.822-0.947),and the value in predicting poor outcomes was 0.8742(95％CI:0.810-0.938)in BAD patients.Conclusion Cerebral perfu-sion impairment,suPAR and Mac-2BP are independent risk factors for END and poor outcome at 90 d in BAD patients.Combined detection can is helpful in evaluating the patient's condition and predicting the prognosis.

Inflammatory bowel disease(IBD)is a chronic nonspecific inflammatory disease of the intestines,mainly including Crohn's disease(CD)and ulcerative colitis(UC).More than 20％of IBD patients progress to colitis-associated colorectal cancer(CAC)through the inflammation-cancer sequence within 30 years of disease onset.Nuclear transcription factor E2-related factor 2(Nrf2)is a transcription factor closely related to IBD and CAC,which not only affects inflammation but also plays a significant role in regulating the"Inflammation-cancer transformation".In this paper,we review the studies on the mechanism of Nrf2 in the"Inflammation-cancer transformation",focusing on the mechanism and dual roles of Nrf2 in oxidative stress,inflammatory factors,immune response,metabolic reprogramming and gene expression,with an aim of providing a reference for further understanding the potential value of its intervention in the human tumor process.

Objective To investigate the effect of E2 signaling in myofibers on muscular macrophage efferocytosis in mice with cardiotoxin-induced acute skeletal muscle injury.Methods Female wild-type C57BL/6 mice with and without ovariectomy and male C57BL/6 mice were given a CTX injection into the anterior tibial muscle to induce acute muscle injury,followed by intramuscular injection of β-estradiol(E2)or 4-hydroxytamoxifen(4-OHT).The changes in serum E2 of the mice were detected using ELISA,and the number,phenotypes,and efferocytosis of the macrophages in the inflammatory exudates and myofiber regeneration and repair were evaluated using immunofluorescence staining and flow cytometry.C2C12 cells were induced to differentiate into mature myotubes,which were treated with IFN-γ for 24 before treatment with β-Estradiol or 4-OHT.The treated myotubes were co-cultured with mouse peritoneal macrophages in a 1:2 ratio,followed by addition of PKH67-labeled apoptotic mouse mononuclear spleen cells induced by UV irradiation,and macrophage efferocytosis was observed using immunofluorescence staining and flow cytometry.Results Compared with the control mice,the female mice with ovariectomy showed significantly increased mononuclear macrophages in the inflammatory exudates,with increased M1 cell percentage,reduced M2 cell percentage and macrophage efferocytosis in the injured muscle,and obviously delayed myofiber regeneration and repair.In the cell co-culture systems,treatment of the myotubes with β-estradiol significantly increased the number and proportion of M2 macrophages and macrophage efferocytosis,while 4-OHT treatment resulted in the opposite changes.Conclusion In injured mouse skeletal muscles,myofiber E2 signaling promotes M1 to M2 transition to increase macrophage efferocytosis,thereby relieving inflammation and promoting muscle regeneration and repair.

Objective To investigate the effect of E2 signaling in myofibers on muscular macrophage efferocytosis in mice with cardiotoxin-induced acute skeletal muscle injury.Methods Female wild-type C57BL/6 mice with and without ovariectomy and male C57BL/6 mice were given a CTX injection into the anterior tibial muscle to induce acute muscle injury,followed by intramuscular injection of β-estradiol(E2)or 4-hydroxytamoxifen(4-OHT).The changes in serum E2 of the mice were detected using ELISA,and the number,phenotypes,and efferocytosis of the macrophages in the inflammatory exudates and myofiber regeneration and repair were evaluated using immunofluorescence staining and flow cytometry.C2C12 cells were induced to differentiate into mature myotubes,which were treated with IFN-γ for 24 before treatment with β-Estradiol or 4-OHT.The treated myotubes were co-cultured with mouse peritoneal macrophages in a 1:2 ratio,followed by addition of PKH67-labeled apoptotic mouse mononuclear spleen cells induced by UV irradiation,and macrophage efferocytosis was observed using immunofluorescence staining and flow cytometry.Results Compared with the control mice,the female mice with ovariectomy showed significantly increased mononuclear macrophages in the inflammatory exudates,with increased M1 cell percentage,reduced M2 cell percentage and macrophage efferocytosis in the injured muscle,and obviously delayed myofiber regeneration and repair.In the cell co-culture systems,treatment of the myotubes with β-estradiol significantly increased the number and proportion of M2 macrophages and macrophage efferocytosis,while 4-OHT treatment resulted in the opposite changes.Conclusion In injured mouse skeletal muscles,myofiber E2 signaling promotes M1 to M2 transition to increase macrophage efferocytosis,thereby relieving inflammation and promoting muscle regeneration and repair.

Objective:To investigate effect of reperfusion injury following different ischemic duration on skeletal muscle in rats.Methods:A model of ischemia/reperfusion injury (IRI) was established by unilateral clamping femoral artery and additional application of tourniquet in skeletal muscle of hind limbs in 35 male Wsitar rats. According to different ischemia time, the animals were assigned to 2-hour ischemia and 24-hour reperfusion (I2R24 group), 2.5-hour ischemia and 24-hour reperfusion (I2.5R24 group), 3-hour ischemia and 24-hour reperfusion (I3R24 group), 4-hour ischemia and 24-hour reperfusion (I4R24 group) and sham group, with 7 rats per group. At the end of reperfusion, gastrocnemious tissues and plasma samples were collected and analyzed. The ratio of wet ∶ dry weight (W/D) was used to measure muscle edema. The assay of 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was conducted to evaluate muscle viability. HE staining was executed to observe histopathological changes. Immunofluorescence staining was performed to assess the levels of C1q, C3b/c, tissue factor (TF), fibrinogen (FN), bradykinin receptor 1 (BR1), BR2, vascular cell adhesion molecule-1 (VCAM-1), E-selectin, fibrinogen-like protein-2 (FGL-2) and myeloperoxidase (MPO) in muscle tissues. ELISA method was used to determine the concentrations of interferonγ (IFN-γ), interleukin7 (IL-7), IL-18, macrophage inflammatory1α (MIP-1α) and monocyte chemotactic protein 1 (MCP-1) in plasma.Results:With prolongation of ischemia time and subsequent reperfusion, tissue edema became severe gradually. The ratio of W/D was 5.3±0.2, 6.1±0.3, 6.9±0.2, 7.6±0.3 in I2R24, I2.5R24, I3R24 and I4R24 groups, higher than that in sham group (4.5±0.1) (all P<0.01). Muscle viability got decreased gradually. Muscle viability was (62.4±3.5)%, (45.3±3.3)%, (35.4±3.4)%, (27.1±5.9)% in I2R24, I2.5R24, I3R24 and I4R24 groups, lower than that in sham group［(93.8±7.2)%］(all P<0.01). Histopathological changes became aggravated gradually. The most severe group was I4R24 group, with the most severe myocyte injury, interstitial edema and extensive inflammatory infiltration, followed by I3R24, I2.5R24 and I2R24 groups in order. There was normal structure integrity and neatly arranged myocyte in sham group. Meanwhile, levels of C1q, C3b, FN, BR1, VCAM-1, E-selectin and FGL-2 got increased gradually. The highest levels for these factors were seen in I4R24 group, followed by I3R24 group, I2.5R24 group, I2R24 group and sham group in order. The rough ratio of the number of positive MPO cells/total cell number under high lens (×200) were increased gradually, with the highest level in I4R24 group, followed by I3R24 group, I2.5R24 group, I2R24 group and sham group in order. However, expression of TF and BR2 were not altered significantly among the groups. Plasma levels of INF-γ, IL-7, IL-18, MIP-1α and MCP-1 elevated gradually with prolongation of ischemia time (all P<0.01). The sequence was the sham group, I2R24 group, I2.5R24 group, I3R24 group and I4R24 groups for levels of these factors from low to high (all P<0.01). Conclusion:Reperfusion after prolongation of ischemia duration can increase the activation of complement, coagulation, kinin and endothelial cells as well as the release of inflammatory factors, and thus aggravate the degree of skeletal muscle tissue injury.

Objective:To investigate the effect of transforming growth factor (TGF- β) signal in muscle fiber itself during inflammation/immunity response on intramuscular inflammation. Methods:Sixteen wild C57BL/6 mice (wild group) and sixteen mice with skeletal muscle-specific deficiency of T βRⅡ (knock-out group) between 4-8 weeks of age were selected for this study. Acute muscle injury in mice was induced by injection of myotoxin cardiotoxin (CTX) into gastrocnemius. The differences in intramuscular inflammation were compared between the wild and knock-out groups on 0, 4, 7 and 10 d after CTX injection by observing exudation of mononuclear phagocytes, macrophages, M1 type macrophages, CD4 +T cells and helpers T cells (Th1, 2&17). Two newborn C57BL/6 wild mice and 2 SM TGF- βr2-/- knock-out mice were selected to culture primary myoblasts in vitro which were divided into 2 groups: an interferon group subjected to interferon simulation and a control group subjected to addition of an equal amount of solvent. The differences in expression of IL-6, IL-10, MCP-1, MIP-1α, H-2K b, H2-Ea, Toll-like receptor (TLR)3 and TLR7 were compared between the interferon and control groups, as well as between the wild and knock-out groups. Results:On 4&7 d after CTX injection, the ratios of mononuclear/macrophage (75.73%±3.62%, 45.27%± 2.32%), macrophages (38.67%±2.76%, 24.87%±2.19%), M1 macrophages (43.21%±0.11%, 30.43%±2.19%), CD4 +T cells (20.13%±1.62%, 5.67%±0.32%) in the muscle tissue from the knock-out mice were significantly higher than those from the wild mice (58.52%±2.43%, 29.21%±2.45%; 20.63%±2.32%, 16.23%±1.25%; 24.98%±0.35%, 14.23%±1.69%; 10.70%±0.43%, 2.50%±0.45%), with a majority of Th1&Th17 ( P<0.05). In vitro results showed that the levels of IL-6, MCP-1, MIP-1α, H-2K b, H2-Ea and TLR3 were significantly upregulated in the interferon group compared with the control group and that such upregulation in the nock-out mice was more significant than in the wild mice ( P<0.05). Conclusions:Endogenous TGF- β signal activation plays a role in the functional recovery after muscle trauma, because it is involved in the regulation of immune behavior of muscle fibers, thus affecting intramuscular inflammation and muscle regeneration.