Objective To explore the effects of long non-coding RNA(lncRNA)HEM2M overexpression on liver injury in mice with non-alcoholic fatty liver disease(NAFLD).Methods Wild-type C57BL/6(WT)mice and myeloid cell-specific HEM2M knock-in(MYKI)mice were fed normal(ND)or high-fat diet(HFD)for 12 weeks.After intraperitoneal glucose tolerance and insulin tolerance tests,the mice were euthanized for detection of liver function indicators in the serum and liver tissue.HE staining and F4/80 immunohistochemical staining were used to examine liver pathologies,and the levels of IL-6,IL-1β,and TNF-α in the liver tissues were determined with ELISA.The mRNA expressions of HEM2M and the markers of M1 macrophages(TNF-α,iNOS,and IL-6)and M2 macrophages(Arg-1,YM-1,and IL-10)were detected using qRT-PCR,and the protein expressions of P-AKT,T-AKT,NLRC4,caspase-1 and GSDMD were assayed using immunoblotting.Caspase-1 activity in the liver tissues was determined with colorimetric measurement and immunofluorescence assay.Results Compared with HFD-fed WT mice,MYKI mice with HFD feeding showed milder liver function damage(P<0.01),alleviated hepatic steatosis,and reduced liver macrophage infiltration,glucose tolerance impairment and insulin resistance(P<0.01).The levels of IL-6,IL-1β,and TNF-α and mRNA expressions of M1 type macrophage markers were significantly decreased(P<0.01)and those of M2 type markers increased(P<0.01)in the liver tissues of HFD-fed MYKI mice,which also showed reduced NLRC4 inflammasome activity,caspase-1 activation,and GSDMD-N protein expression compared with their WT counterparts(P<0.05).Conclusion Overexpression of HEM2M reduces the production of hepatic inflammatory factors,improves insulin resistance and inhibits hepatic NLRC4 inflammasome activation,which leads to reduced hepatic pyroptosis and liver injury in NAFLD mice.

Objective To investigate the role of RNA-binding motif protein X-linked(RBMX)in regulating the proliferation,migration,invasion and glycolysis in human bladder cancer cells.Methods A lentivirus vectors system and RNA interference technique were used to construct bladder cancer 1376 and UC-3 cell models with RBMX overexpression and knockdown,respectively,and successful cell modeling was verified using RT-qPCR and Western blotting.Proliferation and colony forming ability of the cells were evaluated using EdU assay and colony-forming assay,and cell migration and invasion abilities were determined using Transwell experiment.The expressions of glycolysis-related proteins M1 pyruvate kinase(PKM1)and M2 pyruvate kinase(PKM2)were detected using Western blotting.The effects of RBMX overexpression and knockdown on glycolysis in the bladder cancer cells were assessed using glucose and lactic acid detection kits.Results RT-qPCR and Western blotting confirmed successful construction of 1376 and UC-3 cell models with RBMX overexpression and knockdown.RBMX overexpression significantly inhibited the proliferation,clone formation,migration and invasion of bladder cancer cells,while RBMX knockdown produced the opposite effects.Western blotting results showed that RBMX overexpression increased the expression of PKM1 and decreased the expression of PKM2,while RBMX knockdown produced the opposite effects.Glucose consumption and lactate production levels were significantly lowered in the cells with RBMX overexpression(P<0.05)but increased significantly following RBMX knockdown(P<0.05).Conclusion RBMX overexpression inhibits bladder cancer progression and lowers glycolysis level in bladder cancer cells by downregulating PKM2 expression,suggesting the potential of RBMX as a molecular target for diagnosis and treatment of bladder cancer.

Objective To investigate the effect of overexpression of LncRNA MEG3 on proliferation,migration and cisplatin sensitivity of hepatoma cells HepG2 and LM3 and explore the underlying and mechanism.Methods The expression of MEG3 in healthy individuals and patients with hepatocellular carcinoma(HCC)was analyzed by online bioinformatics analysis,and Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect MEG3 expression in different HCC cell lines.A MEG3-overexpresing plasmid was transfected in HepG2 and LM3 cells,and the changes in cell proliferation and migration were examined using CCK8 assay and scratch assay.CCK8 assay was used to determine the inhibitory rate of cisplatin on the transfected cells.A reactive oxygen species(ROS)fluorescence probe(DCFH-DA)and malondialdehyde(MDA)kit were used to assess the changes in ROS production and MDA level in the cells.Western blotting was performed to detect the expression levels of ferroptosis-related proteins glutathione peroxidase 4(GPX4)and ferritin heavy chain 1(FTH1).Results The expression of MEG3 was significantly lower in HCC cells than in LO2 cells,which was consistent with the results of bioinformatic analysis(P<0.05).Overexpression of MEG3 in the HCC cell lines significantly suppressed cell proliferation and migration(P<0.05),increased the cell inhibition rate of cisplatin(P<0.05),enhanced cellular ROS production and increased MDA levels in the cells(P<0.05).MEG3 overexpression significantly decreased the expressions of GPX4 and FTH1 in the HCC cell lines.Conclusion The expression of MEG3 is decreased in HCC cells,and its overexpression inhibits proliferation and migration and enhances cisplatin sensitivity of HCC cells by promoting ferroptosis of the cells.

Objective To elucidate the role of programmed cell death factor 4(PDCD4)in mitochondrial dysfunction caused by sepsis-related vascular endothelial damage.Methods Cultured human umbilical vein endothelial cells(HUVECs)and mouse vascular endothelial cells(C166 cells)were transfected with a small interfering RNA targeting PDCD4 followed by treatment with lipopolysaccharide(LPS)alone or in combination with carbonyl cyanide 3-chlorophenylhydrazone(FCCP).The proteomic changes in the cells after PDCD4 knockdown were analyzed using LC-MS/MS technique.The mRNA expressions of PDCD4 and the genes associated with cell inflammation and apoptosis were detected with RT-PCR,and the expressions of FIS1,DRP1 and OPA1 proteins key to mitochondrial fission and fusion were determined using Western blotting.JC-1 and MitoSOX fluorescent probes were used to observe the changes in mitochondrial membrane potential and mitochondrial reactive oxygen species levels under by a laser confocal microscope.Results LPS stimulation of the cells significantly increased the mRNA expressions of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and monocyte chemoattractant protein 1(MCP1)and enhanced the cellular expression of PDCD4(P<0.05).Proteomic analysis suggested a correlation between PDCD4 knockdown and changes in mitochondrial dynamics in the cells.LPS treatment significantly increased the expressions of mitochondrial fission proteins FIS1 and DRP1 and lowered the expression of the fusion protein OPA1 in the cells(P<0.05),causing also mitochondrial oxidative stress and reduction of the mitochondrial membrane potential(P<0.05).In HUVECs,treatment with FCCP significantly attenuated the protective effect of PDCD4 knockdown,which inhibited LPS-induced inflammation and oxidative stress and restored the balance between mitochondrial fission and fusion.Conclusion PDCD4 knockdown protects vascular endothelial cells against LPS-induced damages by repressing mitochondrial fission and oxidative stress,promoting mitochondrial fusion,and maintaining normal mitochondrial function.

Objective To explore the correlation between polycystic ovary syndrome(PCOS)and periodontitis in light of cytokines levels,sex hormone levels and metabolism-related indicators and their changes during progression of the two diseases.Methods Twenty healthy subjects and 40 patients diagnosed with PCOS underwent full-mouth periodontal examinations to obtain full-mouth plaque score(FMPS),gingival bleeding index of probing(BOP),probing depth(PD),and clinical attachment level(CAL).The participants were divided into Group A without periodontitis or PCOS(n=15),Group B with PCOS but without periodontitis(n=28),Group C with periodontitis but without PCOS(n=5),and Group D with both diseases(n=12).Serum levels of luteinizing hormone/follicle stimulating hormone(LH/FSH),testosterone,prolactin,progesterone and estradiol,and the levels of interleukin 6(IL-6),IL-17A,tumor necrosis factor α and matrix metalloproteinase 8(MMP-8)in both serum and saliva samples were measured at the time of enrolment and at 3 and 6 months after enrolment and compared among the 4 groups.Results Serum MMP-8 level was significantly higher in Group B than in Group A(P<0.05).Salivary MMP-8 level was significantly higher in Group D than in Group B(P<0.05).Salivary MMP-8,LH,and LH/FSH levels and serum and salivary IL-6 and progesterone levels all tended to increase in the 6 months after enrollment(OR>1,P<0.05).During the follow-up period,serum IL-6 levels differed significantly between the non-PCOS groups(A and C)and PCOS groups(B and D)(P<0.05);serum IL-6 and salivary MMP-8 levels differed significantly between the non-periodontitis groups(A and B)and periodontitis groups(C and D)(P<0.05).Spearman correlation analysis indicated positive correlations of LH and LH/FSH with PD(P<0.05);testosterone and LH/FSH were positively correlated with serum MMP-8 levels(P<0.05),and PD,BOP and FMPS were positively correlated with salivary MMP-8 levels(P<0.01).Conclusion There is a correlation between PCOS and periodontitis,and their progression is accompanied by changes in serum and salivary levels of pro-inflammatory cytokines and serum sex hormones.

Objective To investigate whether resveratrol alleviates hyperglycemia-induced cardiomyocyte hypertrophy by enhancing the expression of silent information regulation 2 homolog 1(SIRT1)to maintain mitochondrial homeostasis.Methods Rat cardiomyocytes H9c2 cells with or without lentivirus-mediated mRNA interference of SIRT1 were cultured in high glucose(HG)and treated with resveratrol for 72 h.The changes in superoxide dismutase(SOD)activity,malondialdehyde(MDA)content,reactive oxygen species(ROS)level,and relative surface of the cells were examined,and the mRNA expressions of atrial natriuretic factor(ANF)and brain natriuretic peptide(BNP)and protein expressions of SIRT1,mitochondrial fusion related proteins optic atrophy protein 1(OPA1)and mitofusin 2,mitochondrial division related proteins dynamin-related protein 1(DRP1)and fission protein 1(FIS1),and mitophagy-related proteins BNIP3L and LC3 were detected using RT-qPCR and Western blotting.Results HG exposure significantly decreased SOD activity,increased MDA content,ROS production,relative cell surface,and the mRNA expressions of ANF and BNP in the cardiomyocytes;the protein expressions of SIRT1,OPA1,mitofusin 2 and BNIP3L and LC3-Ⅱ/LC3-Ⅰ ratio were all decreased and the protein expressions of DRP1 and FIS1 increased in HG-exposed cells(P<0.01).All these changes in HG-exposed cardiomyocytes were significantly alleviated by treatment with resveratrol(P<0.05).The protective effects of resveratrol against HG exposure in the cardiomyocytes were obviously attenuated by transfection of the cells with si-SIRT1(P<0.05).Conclusion Resveratrol inhibits hyperglycemia-induced cardiomyocyte hypertrophy by reducing oxidative stress,the mechanisms of which involve enhancement of SIRT1 protein expression,regulation of mitochondrial fusion and division balance,and promoting BNIP3L-mediated mitophagy to maintain mitochondrial homeostasis in the cells.

Objective To investigate the effects of galangin on angiogenic activity of oxidized low-density lipoprotein(ox-LDL)-induced human aortic endothelial cells(HAECs)and explore the underlying mechanisms.Methods HAECs incubated with 10,20,40,and 80 μmol/L galangin for 24 h were assessed for cell viability changes using MTT assay to determine the cytotoxicity of galangin.HAECs treated with 5 mg/mL ox-LDL and incubated with 20 and 40 μmol/L galangin for 24 h,and the cells overexpressing lncRNA H19 and incubated with 40 μmol/L galangin for 24 h were examined for lncRNA H19 level with qRT-PCR.The migration and tube formation capacity of the cells were observed using scratch assay and angiogenesis assay,and ROS levels in the cells were detected with flow cytometry.The protein expression levels of VEGFA,MMP-2 and MMP-9 in the treated cells were detected with Western blotting.Results Galangin at 10,20,or 40 μmol/L produced no obvious toxicity(P>0.05),whereas 80 μmol/L galangin significantly inhibited the viability of HAECs(P<0.01).Treatment with ox-LDL significantly increased the expression of lncRNA H19 in HAECs.Galangin significantly lowered lncRNA H19 expression in ox-LDL-induced HAECs,suppressed cell migration,angiogenesis and ROS production level,and reduced the protein levels of VEGFA,MMP-2 and MMP-9(P<0.01).The effects of galangin were blocked by overexpression of lncRNA H19 in the cardiomyocytes.Conclusion The therapeutic effect of galangin for atherosclerosis is mediated by inhibiting lncRNA H19 expression to reduce ox-LDL-induced migration,oxidative stress,and angiogenesis of HAECs.

Objective To assess the effect of platycodin D(PD)for alleviating pulmonary fibrosis in mice and explore the underlying mechanism.Methods C57BL/6J mouse models of pulmonary fibrosis induced by bleomycin injection into the airway were treated with daily intragastric administration of 10 mg/kg PD for 28 days.The changes of pulmonary fibrosis and the expression and distribution of transient receptor potential cation channel subfamily C member 6(TRPC6)were evaluated with immunohistochemistry,HE staining and Sirius Red staining.Western blotting was used to detect α-SMA expression in the lung tissues of the mice.Primary cultures of mouse lung fibroblasts were pretreated with PD(2.5,5.0,and 10 μmol/L)or larixyl acetate(LA;10 μmol/L)before exposure to 10 ng/mL transforming growth factor-β1(TGF-β1),and the changes in cell survival rate,expressions of collagen I,α-SMA and TRPC6,reactive oxygen species(ROS)production,mitochondrial membrane potential,and cell proliferation capacity were assessed.Network pharmacology analysis was performed to explore the mechanism by which PD alleviated pulmonary fibrosis.Results PD treatment significantly alleviated pulmonary fibrosis and reduced α-SMA expression in BLM-induced mouse models(P<0.05).In TGF-β1-induced primary mouse lung fibroblasts,PD effectively inhibited the cell proliferation,reduced ROS production(P<0.0001),rescued the reduction of mitochondrial membrane potential(P<0.001),and inhibited the expressions of α-SMA and collagenⅠ(P<0.05).Network pharmacology analysis suggested that TRPC6 mediated the effect of PD for alleviating pulmonary fibrosis.Immunohistochemistry showed that PD significantly reduced TRPC6 expression in the lung tissues of BLM-induced mice.In primary mouse lung fibroblasts,PD significantly inhibited TGF-β1-induced TRPC6 expression(P<0.05),and LA treatment obviously lowered the expression levels of TRPC6,α-SMA and collagenⅠ(P<0.05).Conclusion PD alleviated pulmonary fibrosis in mice possibly by down-regulating TRPC6 and reducing ROS production.

Objective To investigate the expression level of tetratricopeptide repeat protein 9A in tumors and its association with the patients'prognosis and immune infiltration.Methods TTC9A expression in different tumor tissues and its association with prognosis,DNA methylation,tumor mutation burden(TMB),and microsatellite instability(MSI)were analyzed based on data from TCGA and GTEx.TIMER and xCell were used to analyze the relationship between TTC9A expression and immune infiltration.Western blotting and RT-qPCR were used to detect the expression of TTC9A in 4 types of cancer cell lines.Results TTC9A expressions were significantly increased in many tumors and down-regulated in a few cancer types(P<0.05).Western blotting and RT-qPCR showed that TTC9A expressions were elevated in lung,colon and liver cancer cells but decreased in bladder cancer cells.In head and neck squamous cell carcinoma,renal clear cell carcinoma,renal papillary cell carcinoma,low-grade glioma,malignant mesothelioma,and endometrial carcinoma tumors,a high expression of TTC9A was strongly correlated with better overall survival(OS),disease-specific survival(DSS),and progression-free interval(PFI)(P<0.05),but was correlated with worse OS,DSS,and PFI in lung adenocarcinoma,pancreatic adenocarcinoma,adrenal carcinoma,and rectal adenocarcinoma(P<0.05).TTC9A hypermethylation was associated with a more favorable prognosis of glioblastoma multiforme,low-grade glioma,uveal melanoma,and ovarian plasmacytoid cystadenocarcinoma(P<0.05)but with poor prognosis of squamous cell carcinoma of the uterine cervix and intracervical adenocarcinoma,squamous cell carcinoma of head and neck,squamous cell carcinoma of the lungs,adrenal carcinoma,and endometrial carcinoma(P<0.05).In most of the cancer types,TTC9A was significantly correlated with the level of immune cell infiltration(P<0.05).Conclusion TTC9A can be used as a prognostic marker for a variety of cancers and is strongly associated with TBM,MSI and immune cell infiltration.

Objective To propose a method for abdominal multi-organ segmentation assisted by multi-phase CT synthesis.Methods Multi-phase CT synthesis for synthesizing high-quality CT images was used to increase the information details for image segmentation.A transformer block was introduced to help to capture long-range semantic information in cooperation with perceptual loss to minimize the differences between the real image and synthesized image.Results The model was trained using multi-phase CT dataset of 526 total cases from Nanfang Hospital.The mean maximum absolute error(MAE)of the synthesized non-contrast CT,venous phase contrast-enhanced CT(CECT),and delay phase CECT images from arterial phase CECT was 19.192±3.381,20.140±2.676 and 22.538±2.874,respectively,which were better than those of images synthesized using other methods.Validation of the multi-phase CT synthesis-assisted abdominal multi-organ segmentation method showed an average dice coefficient of 0.847 for the internal validation set and 0.823 for the external validation set.Conclusion The propose method is capable of synthesizing high-quality multi-phase CT images to effectively reduce the errors in registration between different phase CT images and improve the performance for segmentation of 13 abdominal organs.