Mitochondria are organelles in eukaryotic cells that play a crucial role in cellular energy me-tabolism,oxidative stress,heat production,and signal transduction.Mitochondrial transplantation(MT)is currently one of the most advanced techniques for treating mitochondrial dysfunction and anti-aging re-search.This study aimed to explore the feasibility and effectiveness of xenogeneic MT by transplanting mitochondria from yak(Bos grunniens),domestic cattle(Bos taurus),and horse(Equus caballus)into mice(Mus musculus).The results demonstrated that mitochondria from yak,domestic cattle,and horse could be successfully transplanted into mice and maintained in various tissues and organs of the mice for at least 14 days,as confirmed by confocal imaging,digital PCR,and DNA sequencing.MT mice exhibi-ted positive biological effects,including increased ATP content and mitochondrial DNA copy number(P＜0.05),with the maximum effect observed on day 7,which was sustained until day 14.Reactive oxygen species(ROS)levels in MT mice significantly increased at 2 hours post-injection(P＜0.05),then grad-ually decreased towards baseline levels by day 7 and day 14(P＞0.05).These findings support the effec-tiveness of xenogeneic MT and suggest that the effects can be maintained for up to 14 days.This study provides scientific evidence for future clinical applications.

Objective:To explore the clinical value of plasma von Willebrand factor antigen(VWF:Ag)and VWF activity(VWF:GPIbM)based on ABO blood group in the risk assessment of deep vein thrombosis(DVT).Methods:A total of 163 patients with DVT who sought medical treatment from March 2021 to December 2022 were selected as the case group,and 135 healthy volunteers during the same period were selected as the control group.The differences of ABO blood groups,plasma VWF:Ag and VWF:GPIbM levels between the two groups were compared.Receiver operating characteristic(ROC)curves were used to evaluate the clinical value of VWF testing in predicting DVT events.Logistic regression analysis was applied to identify risk factors for DVT.Results:The levels of plasma VWF:Ag and VWF:GPIbM in the DVT group were significantly higher than those in the control group both overall and across ABO blood type subgroups(P＜0.01).Within the DVT group,the levels of plasma VWF:Ag and VWF:GPIbM in patients with non-O blood type were significantly higher than those with blood type O[VWF:Ag:219.74％±63.64％vs 162.21％±56.03％,P＜0.01;VWF:GPIbM:228.10％(185.15％,249.10％)vs 148.25％(116.48％,225.48％),P＜0.01].The area under the ROC curve(AUC)of VWF:Ag for predicting DVT events was 0.855,with a cut-off value of 142.4％,sensitivity of 82.2％and specificity of 72.6％;the AUC of VWF:GPIbM was 0.861,with a cut-off value of 141.2％,sensitivity of 84.7％,and specificity of 71.1％.Univariate analysis showed that both VWF:Ag and VWF:GPIbM were influencing factors for DVT events(P＜0.05).Multivariate logistic regression analysis indicated that VWF:Ag＞142.4％(OR=13.961,95％CI:7.654-25.464,P＜0.01)and VWF:GPIbM＞141.2％(OR=17.615,95％CI:9.155-33.892,P＜0.01)were independent risk factors for DVT events.Conclusion:Levels of VWF:Ag and VWF:GPIbM are significantly elevated in non-O blood type DVT patients.VWF:Ag＞142.4％and VWF:GPIbM＞141.2％are independent risk factors for DVT events.VWF testing based on ABO blood group aids in the precision prevention and control of DVT.

OBJECTIVE: To investigate the effect of miR-483-5p on hepatocellular carcinoma (HCC) cells proliferation and stemness, as well as the attenuating effect of Pien Tze Huang (PZH). METHODS: Differentially expressed miRNA between HepG2 cells and hepatic cancer stem-like cells (HCSCs) were identified by a miRNA microarray assay. miR-483-5p mimics were transfected into HepG2 cells to explore the effects of miR-483-5p on cell proliferation and stemness. HepG2 cells and HCSCs were treated with PZH (0, 0.25, 0.50 and 0.75 mg/mL) to explore the effects of PZH on the proliferation and stemness, both in non-induced state and the state induced by miR-483-5p mimics. RESULTS: miR-483-5p was significantly up-regulated in HCSCs and its overexpression increased cell proliferation and stemness in HepG2 cells (P<0.05). PZH not only significantly inhibited proliferation in HepG2 cells, but also significantly suppressed the cell proliferation and self-renewal of HCSCs (P<0.05). The effects of miR-483-5p mimics on proliferation and stemness of HepG2 cells were partially abolished by PZH. CONCLUSIONS miR-483-5p promotes proliferation and enhances stemness of HepG2 cells, which were attenuated by PZH, demonstrating that miR-483-5p is a potential molecular target for the treatment of HCC and provide experimental evidence to support clinical use of PZH for patients with HCC.
Humans ; MicroRNAs/metabolism* ; Cell Proliferation/drug effects* ; Liver Neoplasms/drug therapy* ; Carcinoma, Hepatocellular/drug therapy* ; Hep G2 Cells ; Neoplastic Stem Cells/metabolism* ; Drugs, Chinese Herbal/therapeutic use* ; Gene Expression Regulation, Neoplastic/drug effects*

Aim To explore the mechanism of the syn-ergistic effect of the ferroptosis inducer erastin com-bined with shikonin in promoting ferroptosis in human hypertrophic scar fibroblasts(HSFBs).Methods Hypertrophic scar tissues provided by the General Hos-pital of Ningxia Medical University were collected,and HSFBs were extracted.HSFBs were identified by HE staining and immunofluorescence.The inhibitory rates of Era and SHK on HSFBs at different concentrations were detected by CCK-8 assay,and the IC50 value was calculated.CompuSyn software was used to calculate the co-use index(CI).Control group,Erastin(Era)group,shikonin(SHK)group and Era+SHK group were set up,and the number and morphological chan-ges of cells were observed after 24 hours of interven-tion.The ability of cell migration and invasion was de-tected by scratch test and Transwell test.The changes of malondialdehyde(MDA),total iron ion and reactive oxygen species(ROS)were detected by corresponding biochemical kits.The expressions of collagen I,α-SMA and GOT1,SLC7A11,GPX4 and FTH1 were detected by Western blot.Results The IC50 value of Era and SHK of primary HSFBs was 2.22 μmol·L-1 and 3.94μmol·L-1 respectively,which was used as the single drug concentration for subsequent experiments.The CompuSyn software was employed to calculate the CI value when the two drugs were used in combination,and the concentrations corresponding to CI=0.39597(Era:1.2 μmol·L-1+SHK:1.5 μmol·L-1)were selected as subsequent combination concentrations(Because when CI was equal to 0.395 97,the concen-tration of each drug was lower than the concentration of single drug,and the inhibition rate of combined drug was greater than 50％).Compared with the monother-apy group,the number of HSFBs in the SHK+Era group was significantly reduced,cell membrane showed breakage and vesiculation,cell wrinkling became smal-ler,and cytoplasm was concentrated.The migration and invasion ability of HSFBs in the SHK+Era group were obviously weakened(P＜0.05),and the expres-sion of fibrosis-related proteins collagen Ⅰ and α-SMA was reduced(P＜0.05);the contents of MDA,total i-ron ions,and ROS in HSFBs of the SHK+Era group increased(P＜0.05),and the protein expression lev-els of SLC7A11,GOT1,GPX4,and FTH1 further de-creased(P＜0.05).Conclusions Erastin in combi-nation with shikonin can synergistically inhibit the pro-liferation,migration and fibrosis levels of HSFBs.The mechanism may be that erastin enhances the inhibition of shikotin on GOT1,increases the levels of cellular i-ron ions,ROS,and lipid peroxides,thereby promoting ferroptosis in HSFBs.

Aim To investigate the effect of metformin on epithelial-mesenchymal transition(EMT)of lung cancer A549 cells and its underlying mechanism.Methods Lung cancer A549 cells were cultured in vitro and treated with metformin.Cell morphology was observed by fluorescence staining.The mRNA expres-sion levels of E-cadherin,N-cadherin,SMA and Vimen-tin were detected by RT-PCR.The regulatory effects of metformin on EMT in A549 cellswere examined by high-throughput sequencing.An EMT model was estab-lished through TGF-β1 induction.Following metformin treatment,the morphology of A549 cells was observed.Western blot was employed to determine the expression levels of NGF,E-cadherin,N-cadherin,SMA and Vim-entin.Additionally,si-NGF transfection was performed to evaluate the protein expressions of E-cadherin,N-cadherin,SMA and Vimentin in A549 cells,and a cell scratch assay was conducted to assess cell migration.Results After metformin treatment,A549 cells exhibi-ted a loss of mesenchymal-like morphology,character-ized by a transition to a round shape,a reduction in colony formation,and decreased adherence.RT-PCR and high-throughput sequencing revealed a down-regu-lation in the expression of genes associated with mesen-chymal transition,including N-cadherin,SMA,and Vim-entin,and an up-regulation in the expression of genes associated with epithelial transformation,such as ZO-1 and E-cadherin.Additionally,the expression of nerve growth factor(NGF)was significantly up-regulated.Following transfection with si-NGF,A549 cells treated with metformin exhibited a down-regulation in the ex-pression of the epithelial marker E-cadherin,concomi-tant with an up-regulation in the expression of stromal markers N-cadherin,Vimentin,and SMA.Conclusions Metformin can up-regulate the expression of E-cad-herin and down-regulate the expression of N-cadherin,Vimentin and SMA in lung cancer A549 cells,thereby inhibiting EMT.Additionally,NGF signaling molecules may play a significant role in this process.

Objective To investigate whether successful percutaneous coronary intervention(PCI)could improve symptoms and quality of life(QOL)in left main disease and/or 3-vessel disease patients with diabetes.Methods Patients with left main disease and/or 3-vessel disease who underwent PCI in the First Affiliated Hospital of Air Force Medical University from April 2018 to May 2021 were consecutively enrolled and subdivided into 2 groups:diabetes and no diabetes.Detailed baseline characteristics,symptoms,including dyspnea and angina,assessed with the Rose dyspnea scale(RDS),Seattle angina questionnaire(SAQ),the European quality of life-5 dimensions(EQ-5D)and 12-item short-form health survey(SF-12)questionnaire respectively,procedural details,and 1 month and 1 year follow-up data were collected.Results Among 440 left main disease and/or 3-vessel disease patients,disease was present in 176(40.00％),who had more hypertension,peripheral artery disease,and LCX lesion(all P＜0.05).The incidence of major adverse cardiovascular events(MACE)and all-cause mortality were similar between the two groups(both P＞0.05)at 1 month follow-up,while all-cause mortality in diabetes patients was significantly higher than those without diabetes at 1 year follow-up(P=0.013).Low left ventricular ejection fraction was an independent risk factor for MACE and all-cause mortality at 1 month and 1 year follow-up after successful revascularization(all P＜0.05).Most importantly,symptoms,including dyspnea and angina,and QOL were markedly improved regardless of diabetes both at 1 month and 1 year follow-up(all P＜0.05).Diabetes patients showed improved dyspnea and QOL at similar degree to the non-diabetes patients(all P＞0.05)and a more significantly relieved angina(P=0.013).Additionally,the number of chronic total occlusion(CTO)per patient was identified as an independent risk factor of dyspnea(OR 0.723,95％CI 0.525～0.997,P=0.048)and angina relief(OR 0.686,95％CI 0.473～0.995,P=0.047),and the contrast volume(OR 0.995,95％CI 0.992～0.999,P=0.008)as an independent risk factor of QOL improvement in diabetic patients.Conclusions Successful PCI is beneficial for relieving symptoms and improving quality of life in patients with diabetes who have left main disease and/or 3-vessel disease.

Objective To investigate whether successful percutaneous coronary intervention(PCI)could improve symptoms and quality of life(QOL)in left main disease and/or 3-vessel disease patients with diabetes.Methods Patients with left main disease and/or 3-vessel disease who underwent PCI in the First Affiliated Hospital of Air Force Medical University from April 2018 to May 2021 were consecutively enrolled and subdivided into 2 groups:diabetes and no diabetes.Detailed baseline characteristics,symptoms,including dyspnea and angina,assessed with the Rose dyspnea scale(RDS),Seattle angina questionnaire(SAQ),the European quality of life-5 dimensions(EQ-5D)and 12-item short-form health survey(SF-12)questionnaire respectively,procedural details,and 1 month and 1 year follow-up data were collected.Results Among 440 left main disease and/or 3-vessel disease patients,disease was present in 176(40.00％),who had more hypertension,peripheral artery disease,and LCX lesion(all P＜0.05).The incidence of major adverse cardiovascular events(MACE)and all-cause mortality were similar between the two groups(both P＞0.05)at 1 month follow-up,while all-cause mortality in diabetes patients was significantly higher than those without diabetes at 1 year follow-up(P=0.013).Low left ventricular ejection fraction was an independent risk factor for MACE and all-cause mortality at 1 month and 1 year follow-up after successful revascularization(all P＜0.05).Most importantly,symptoms,including dyspnea and angina,and QOL were markedly improved regardless of diabetes both at 1 month and 1 year follow-up(all P＜0.05).Diabetes patients showed improved dyspnea and QOL at similar degree to the non-diabetes patients(all P＞0.05)and a more significantly relieved angina(P=0.013).Additionally,the number of chronic total occlusion(CTO)per patient was identified as an independent risk factor of dyspnea(OR 0.723,95％CI 0.525～0.997,P=0.048)and angina relief(OR 0.686,95％CI 0.473～0.995,P=0.047),and the contrast volume(OR 0.995,95％CI 0.992～0.999,P=0.008)as an independent risk factor of QOL improvement in diabetic patients.Conclusions Successful PCI is beneficial for relieving symptoms and improving quality of life in patients with diabetes who have left main disease and/or 3-vessel disease.

Objective:To explore the clinical value of plasma von Willebrand factor antigen(VWF:Ag)and VWF activity(VWF:GPIbM)based on ABO blood group in the risk assessment of deep vein thrombosis(DVT).Methods:A total of 163 patients with DVT who sought medical treatment from March 2021 to December 2022 were selected as the case group,and 135 healthy volunteers during the same period were selected as the control group.The differences of ABO blood groups,plasma VWF:Ag and VWF:GPIbM levels between the two groups were compared.Receiver operating characteristic(ROC)curves were used to evaluate the clinical value of VWF testing in predicting DVT events.Logistic regression analysis was applied to identify risk factors for DVT.Results:The levels of plasma VWF:Ag and VWF:GPIbM in the DVT group were significantly higher than those in the control group both overall and across ABO blood type subgroups(P＜0.01).Within the DVT group,the levels of plasma VWF:Ag and VWF:GPIbM in patients with non-O blood type were significantly higher than those with blood type O[VWF:Ag:219.74％±63.64％vs 162.21％±56.03％,P＜0.01;VWF:GPIbM:228.10％(185.15％,249.10％)vs 148.25％(116.48％,225.48％),P＜0.01].The area under the ROC curve(AUC)of VWF:Ag for predicting DVT events was 0.855,with a cut-off value of 142.4％,sensitivity of 82.2％and specificity of 72.6％;the AUC of VWF:GPIbM was 0.861,with a cut-off value of 141.2％,sensitivity of 84.7％,and specificity of 71.1％.Univariate analysis showed that both VWF:Ag and VWF:GPIbM were influencing factors for DVT events(P＜0.05).Multivariate logistic regression analysis indicated that VWF:Ag＞142.4％(OR=13.961,95％CI:7.654-25.464,P＜0.01)and VWF:GPIbM＞141.2％(OR=17.615,95％CI:9.155-33.892,P＜0.01)were independent risk factors for DVT events.Conclusion:Levels of VWF:Ag and VWF:GPIbM are significantly elevated in non-O blood type DVT patients.VWF:Ag＞142.4％and VWF:GPIbM＞141.2％are independent risk factors for DVT events.VWF testing based on ABO blood group aids in the precision prevention and control of DVT.

Aim To explore the mechanism of the syn-ergistic effect of the ferroptosis inducer erastin com-bined with shikonin in promoting ferroptosis in human hypertrophic scar fibroblasts(HSFBs).Methods Hypertrophic scar tissues provided by the General Hos-pital of Ningxia Medical University were collected,and HSFBs were extracted.HSFBs were identified by HE staining and immunofluorescence.The inhibitory rates of Era and SHK on HSFBs at different concentrations were detected by CCK-8 assay,and the IC50 value was calculated.CompuSyn software was used to calculate the co-use index(CI).Control group,Erastin(Era)group,shikonin(SHK)group and Era+SHK group were set up,and the number and morphological chan-ges of cells were observed after 24 hours of interven-tion.The ability of cell migration and invasion was de-tected by scratch test and Transwell test.The changes of malondialdehyde(MDA),total iron ion and reactive oxygen species(ROS)were detected by corresponding biochemical kits.The expressions of collagen I,α-SMA and GOT1,SLC7A11,GPX4 and FTH1 were detected by Western blot.Results The IC50 value of Era and SHK of primary HSFBs was 2.22 μmol·L-1 and 3.94μmol·L-1 respectively,which was used as the single drug concentration for subsequent experiments.The CompuSyn software was employed to calculate the CI value when the two drugs were used in combination,and the concentrations corresponding to CI=0.39597(Era:1.2 μmol·L-1+SHK:1.5 μmol·L-1)were selected as subsequent combination concentrations(Because when CI was equal to 0.395 97,the concen-tration of each drug was lower than the concentration of single drug,and the inhibition rate of combined drug was greater than 50％).Compared with the monother-apy group,the number of HSFBs in the SHK+Era group was significantly reduced,cell membrane showed breakage and vesiculation,cell wrinkling became smal-ler,and cytoplasm was concentrated.The migration and invasion ability of HSFBs in the SHK+Era group were obviously weakened(P＜0.05),and the expres-sion of fibrosis-related proteins collagen Ⅰ and α-SMA was reduced(P＜0.05);the contents of MDA,total i-ron ions,and ROS in HSFBs of the SHK+Era group increased(P＜0.05),and the protein expression lev-els of SLC7A11,GOT1,GPX4,and FTH1 further de-creased(P＜0.05).Conclusions Erastin in combi-nation with shikonin can synergistically inhibit the pro-liferation,migration and fibrosis levels of HSFBs.The mechanism may be that erastin enhances the inhibition of shikotin on GOT1,increases the levels of cellular i-ron ions,ROS,and lipid peroxides,thereby promoting ferroptosis in HSFBs.

Aim To investigate the effect of metformin on epithelial-mesenchymal transition(EMT)of lung cancer A549 cells and its underlying mechanism.Methods Lung cancer A549 cells were cultured in vitro and treated with metformin.Cell morphology was observed by fluorescence staining.The mRNA expres-sion levels of E-cadherin,N-cadherin,SMA and Vimen-tin were detected by RT-PCR.The regulatory effects of metformin on EMT in A549 cellswere examined by high-throughput sequencing.An EMT model was estab-lished through TGF-β1 induction.Following metformin treatment,the morphology of A549 cells was observed.Western blot was employed to determine the expression levels of NGF,E-cadherin,N-cadherin,SMA and Vim-entin.Additionally,si-NGF transfection was performed to evaluate the protein expressions of E-cadherin,N-cadherin,SMA and Vimentin in A549 cells,and a cell scratch assay was conducted to assess cell migration.Results After metformin treatment,A549 cells exhibi-ted a loss of mesenchymal-like morphology,character-ized by a transition to a round shape,a reduction in colony formation,and decreased adherence.RT-PCR and high-throughput sequencing revealed a down-regu-lation in the expression of genes associated with mesen-chymal transition,including N-cadherin,SMA,and Vim-entin,and an up-regulation in the expression of genes associated with epithelial transformation,such as ZO-1 and E-cadherin.Additionally,the expression of nerve growth factor(NGF)was significantly up-regulated.Following transfection with si-NGF,A549 cells treated with metformin exhibited a down-regulation in the ex-pression of the epithelial marker E-cadherin,concomi-tant with an up-regulation in the expression of stromal markers N-cadherin,Vimentin,and SMA.Conclusions Metformin can up-regulate the expression of E-cad-herin and down-regulate the expression of N-cadherin,Vimentin and SMA in lung cancer A549 cells,thereby inhibiting EMT.Additionally,NGF signaling molecules may play a significant role in this process.