Objective: To investigate the regulatory relationship between long non-coding RNA LINC01578 and c-Myc, and to explore the effect of LINC01578 on the metabolic process of oral squamous cell carcinoma(OSCC). Methods: After c-Myc was knocked down in OSCC cell line CAL27, LINC01578, a long non-coding RNA that is positively regulated by c-Myc, was identified by high-throughput sequencing technology. qRT-PCR was employed to measure the expression levels of c-Myc and LINC01578 in OSCC tissues and adjacent normal tissues. Following overexpression or knockdown of c-Myc in CAL27 and HN6 cells, qRT-PCR was conducted to validate the consistency with sequencing results. The binding of c-Myc to the LINC01578 promoter was confirmed using a dual luciferase reporter assay. Seahorse, ATP production and lactate production assays were utilized to examine the impact of c-Myc on glucose metabolism in OSCC via LINC01578. Colony formation assays assessed the proliferative capacity of OSCC cell lines. Results: qRT-PCR analysis revealed significantly higher expression levels of c-Myc LINC01578 in OSCC tissues compared to adjacent tissues( P < 0. 05 ) , confirming that c⁃Myc positively regulates LINC01578 expression. Consistent with sequencing data , c⁃Myc overexpression markedly upregulated LINC01578 (P < 0. 001) , while c⁃Myc knockdown led to a significant decrease in LINC01578 levels(P < 0. 000 1) . Dual lu ciferase reporter gene assays demonstrated that c⁃Myc directly targets and transcriptionally enhanced LINC01578 ex⁃ pression(P < 0. 001) . Seahorse experiments indicated that c⁃Myc promoted glucose metabolism in OSCC through LINC01578 regulation(P < 0. 05) . Colony formation assays showed that LINC01578 overexpression enhanced OS⁃ CC cell proliferation , whereas LINC01578 knockdown inhibited it. Conclusion c⁃Myc upregulates LINC01578 expression in OSCC cells , thereby modulating glycolysis and promoting cell proliferation.

Objective: To explore the role of leukemia inhibitory factor (LIF) in dental pulp inflammation . Methods: Human dental pulp stem cells (hDPSCs) were cultured in vitro as the target cells , the inflammatory response was induced by lipopolysaccharide (LPS) , and high-throughput sequencing was used to detect relevant highly ex- pressed genes in the inflammatory state . The expression of LIF under graded concentrations of LPS stimulation was detected by real-time fluorescence quantitative PCR (qPCR) . The expression levels of interleukin-6 (IL-6) , inter- leukin-1β(IL-1β) , and tumor necrosis factor-α(TNF-α) were detected after LIF knockdown and overexpression in hDPSCs by qPCR . Normal and inflammatory pulp tissues were collected , and the expression of LIF in both tis- sues was detected by qPCR and immunofluorescence (IF) . Results: The expression level of LIF increased in hu- man dental pulp cells after LPS stimulation . The expression level of LIF was subsequently elevated in inflammatory pulp induced by graded concentrations of LPS . The expression of IL-6 , IL-1β, and TNF-αwas significantly down- regulated in hDPSCs after LIF knockdown in response to LPS stimulation , while LIF overexpression upregulated the expression of these cytokines . qPCR and IF assays showed high expression of LIF in inflamed pulp tissue . Conclusion LIF is involved in dental pulp inflammation and promotes the development of pulpitis .

Objective: To elucidate the molecular mechanism by which prohibitin 2(PHB2) mediates periodontitis-induced bone tissue inflammation through regulating the nuclear factor kappa B(NF-κB) signaling pathway and its role in irreversible alveolar bone resorption. Methods: Quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR) and immunohistochemistry(IHC) were used to detect the expression differences of inflammatory factors and PHB2 in healthy and inflamed alveolar bone tissues of mice in vivo. In vitro, an inflammatory model was established using lipopolysaccharide(LPS)-induced a mouse calvaria-derived preosteoblastic cell line, subclone E1(MC3T3-E1) cells. Western blot and qRT-PCR were used to clarify the regulatory relationship between PHB2 and inflammatory factors, and immunofluorescence staining was performed to observe changes in PHB2 subcellular localization. PHB2 overexpression plasmids were constructed using molecular cloning, and RNA interference was employed to knock down PHB2 expression to assess its regulatory role in inflammation. Based on RNA-seq data, differential expression analysis based on the negative binomial distribution, version 2(DESeq2) was used for differential expression analysis, and kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment along with gene ontology(GO) functional annotation were performed to identify key signaling pathways and differentially expressed genes. Results: In the mouse periodontitis model, PHB2 expression was significantly upregulated in alveolar bone tissues. In the in vitro inflammatory cell model, PHB2 levels positively correlated with interleukin(IL)-6, IL-1β, and tumor necrosis factor-alpha(TNF-α) levels, and its subcellular localization shifted during inflammation. RNA-seq data and the detection of the level of phosphorylation of p65 protein(p-p65) demonstrated that PHB2 exacerbated inflammatory responses through the NF-κB signaling pathway and was mechanistically linked to upregulation of the upstream chemokine C-X-C motif chemokine ligand 10(CXCL10). Conclusion PHB2 aggravates LPS-induced periodontitis inflammation via the NF-κB signaling pathway, providing new insights into the molecular mechanisms underlying the development of periodontitis.

Objective To investigate changes in serum complement C1 tumor necrosis factor-related pro-tein family 12(CTRP12)level before and after percutaneous coronary intervention(PCI)in patients with acute myocardial infarction(AMI)and the relationship of CTRP12 level with in-stent restenosis(ISR).Methods A total of 104 patients who had been diagnosed with AMI and had undergone PCI at Danyang People's Hospital in Jiangsu Province from January 2021 to June 2023 were selected.The incidence of ISR within 12 months after PCI was counted,and they were divided into an ISR group and a non-ISR group according to the results of reviewed coronary angiography.Serum CTRP12 levels were compared between the two groups before PCI and on one day before discharge.Logistic regression was used to analyze the influencing factors of ISR in AMI patients after PCI.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of CTRP12 for ISR in AMI patients after PCI.Results The incidence of ISR in 104 AMI patients at 12 months after PCI was 14.4%(15/104).As compared with the non-ISR group,the ISR group had significant increases in preoperative TIMI flow of≤1,white blood cell count,neutrophil count,TC,and LDL-C,and a significant decline in serum CTRP12 level on one day before discharge(P<0.05).In the non-ISR group,serum CTRP12 level was significantly higher on one day before discharge than its baseline(P<0.05).In the ISR group,serum CTRP12 level on one day before discharge was lower than its baseline,but the difference was not statistically significant(P>0.05).Logistic regression analysis showed that a lower CTRP12 level on one day before discharge was an independent risk factor for ISR in AMI patients after PCI(P<0.05).ROC curve analysis showed that the optimal cut-off point of serum CTRP12 on one day before discharge for predicting ISR in AMI patients after PCI was 3.89 ng/mL(sensitivity 93.3%and speci-ficity 73.0%),and the area under the ROC curve(AUC)was 0.849.Conclusions Serum CTRP12 level inone day before discharge has certain predictive value for ISR in AMI patients after PCI.CTRP12 may be a therapeutic target for ISR in AMI patients after PCI.

Objective This study examined the influence of c-Myc regulated long noncoding RNA TBL1XR1-5(lnc-TBL1XR1-5)on the progression of oral squamous cell carcinoma(OSCC).Methods Based on the expres-sion of c-Myc,bioinformatics analysis was performed on head and neck squamous cell carcinoma samples in the TCGA database,and lnc-TBL1XR1-5 with closely related c-Myc expression was screened.The effects of c-Myc overexpression and knockdown on lnc-TBL1XR1-5 were detected.Expression relationship of c-Myc and lnc-TBL1XR1-5 in OSCC and para-cancer tissues.Dual-luciferase reporter assays were used to verify the binding of c-Myc to the lnc-TBL1XR1-5 promoter region.RNA FISH were used to determine the localization of lnc-TBL1XR1-5 in OSCC cell lines.The effects of overexpression,knockdown of lnc-TBL1XR1-5 on migration,metabolism,and proliferation of OSCC cells were observed by quantitative real-time polymerase chain reaction(qRT-PCR),scratch tests,transwell assays,medium color and pH changes,cell counts,CCK-8 assay,and colony formation assays.Results It was found that c-Myc positively regulated the expression of lnc-TBL1XR1-5 in OSCC cell lines and tis-sues.The binding of c-Myc to the lnc-TBL1XR1-5 promoter region was verified by dual-luciferase reporter assays.RNA FISH showed that lnc-TBL1XR1-5 was localized in the nuclei in OSCC cells.Overexpression of lnc-TBL1XR1-5 promoted migration,metabolism and proliferation in OSCC cell lines,while knockdown of it had the opposite effect.Conclusion c-Myc positively regulates lnc-TBL1XR1-5 in OSCC.Lnc-TBL1XR1-5 affect the mi-gration,proliferation,and metabolism of OSCC by influencing the tumor microenvironment.

Objective To explore the action mechanism of long non-coding RNA(lncRNAs)lncRNA KCTD13-DT in oral squamous cell carcinoma(OSCC)and its potential interaction with transcription factor c-Myc,providing a potential diagnostic and therapeutic target for patients with OSCC.Methods The expression of lncRNA KCTD13-DT in OSCC and paracancerous tissues was detected by qRT-PCR.The effects of c-Myc overexpression and knock-down on human tongue squamous carcinoma cells HN6 and CAL27 were detected by qRT-PCR.Fluorescence in si-tu hybridization(FISH)assessed the localization of lncRNA KCTD13-DT in cells.A dual luciferase reporter gene was used to analyze the role of c-Myc in target binding to the promoter region of lncRNA KCTD13-DT.Stable cell lines with knockdown or overexpression of lncRNA KCTD13-DT were constructed in human OSCC cell lines HN6 and CAL27 by lentiviral infection,and the knockdown and overexpression efficiencies of lncRNA KCTD13-DT were detected by qRT-PCR.Cell proliferation changes were detected by growth curve assay,CCK-8 assay,colony forma-tion assay,and cell migration was detected by scratch assay and Transwell.Results lncRNA KCTD13-DT expres-sion level was reduced in OSCC tissues and OSCC cells(HN6,CAL27),and Western blot verified that after knoc-king down and overexpression of c-Myc in HN6 and CAL27,the qRT-PCR experiments showed that c-Myc nega-tively regulated lncRNA KCTD13-DT,and overexpression of c-Myc significantly down-regulated lncRNA KCTD13-DT;knockdown of c-Myc significantly up-regulated lncRNA KCTD13-DT levels.Dual luciferase reporter gene showed that c-Myc could target lncRNA KCTD13-DT,and c-Myc could be involved in regulating and repressing the transcriptional activity of lncRNA KCTD13-DT.FISH showed that lncRNA KCTD13-DT mainly existed in the nu-cleus.Growth curve assay,CCK-8 assay,cell scratch assay,Transwell,and colony formation assay showed that knockdown of lncRNA KCTD13-DT promoted the growth and proliferation of OSCC cells,and overexpression of ln-cRNA KCTD13-DT significantly inhibited the proliferation and migration of OSCC cells.Conclusion lncRNA KCTD13-DT is negatively regulated by c-Myc.Knockdown of lncRNA KCTD13-DT promotes cell proliferation,while overexpression of it inhibits cell growth.

Cuproptosis is a new type of cell death that depends on intracellular copper accumulation to trigger the aggregation of mitochondrial lipoacylated protein and the degradation of iron-sulfur cluster protein,with a different mechanism of action from autophagy,ferroptosis,pyroptosis,and necroptosis.Cuproptosis is closely association with the development of liver cancer and resistance to antitumor drugs,as well as the progression of various liver diseases such as hereditary liver diseases,nonalcoholic fatty liver disease,viral hepatitis,and liver cirrhosis.This article summarizes the mechanism of cuproptosis and its role in liver diseases,in order to provide a reference for further research and treatment of liver diseases.

Objective To investigate the changes of C1q tumor necrosis factor-related protein-12(CTRP12)levels in serum of the pa-tients with acute myocardial infarction(AMI)before and after percutaneous coronary intervention(PCI),and explore its clinical sig-nificance.Methods A total of 50 patients with AMI who underwent emergency PCI and 35 patients with normal coronary angiography results in Danyang People's Hospital from November 2021 to October 2022 were enrolled.The CTRP12 levels in peripheral venous ser-um were compared between the two groups.The levels of serum CTRP12 levels were measured before,during and on the 3rd,5th and 7th day after PCI.The serum CTRP12 levels in culprit coronary ostium and peripheral vein were compared.CTRP12 levels in peripher-al venous serum were compared at different time points after PCI.The severity of coronary artery disease was evaluated by SYNTAX score system,and the AMI patients were divided into two groups:SYNTAX score ≤22 and SYNTAX score＞22.The serum CTRP12 levels were compared between the two groups and before and after PCI.The correlation between CTRP12 and age,body mass index(BMI),fasting blood glucose,blood lipid and other factors was analyzed.The influencing factors of the severity of coronary artery le-sions were analyzed by logistic regression.Results The serum CTRP12 level in the patients with AMI was significantly lower than that in healthy controls(P＜0.05).There was no significant difference between the serum CTRP12 levels between preoperative peripheral vein and intraoperative culprit coronary orifice(P＞0.05).Compared with that before PCI,the serum CTRP12 level was lower on the 3rd day after PCI(P＜0.05),and increased on the 5th and 7th days after PCI,but no statistically significant difference was found(P＞0.05).Compared with those on the 3rd day after PCI,the serum CTRP12 levels were increased on the 5th and 7th day after PCI,but no statistically significant differences were found(all P＞0.05).Compared with that in the SYNTAX≤22 group,the CTRP12 levels were significantly lower than those before PCI and on the 3rd day after PCI(all P＜0.05),while there was no significant difference on the 5th and 7th day after PCI in SYNTAX＞22 group(all P＞0.05).CTRP12 was negatively correlated with the level of total cholesterol(TC)and positively correlated with high-density lipoprotein cholesterol(HDL-C).Univariate logistic regression analysis showed that CTRP12 was an independent influencing factor for the severity of coronary artery disease in the patients with AMI(β=-1.671,OR=0.188,P＜0.05).After adjusting for the effects of age,gender,BMI,smoking,hypertension,diabetes,fasting blood glucose,total cholesterol(TC),triglyceride(TG),HDL-C and low-density lipoprotein cholesterol(LDL-C),CTRP12 was still an independent in-fluencing factor for the severity of coronary artery disease in the patients with AMI(β=-3.441,OR=0.032,P＜0.05).Conclusion The serum CTRP12 level was significantly decreased in the patients with AMI before PCI,and showed continuous decline on the 3rd day after PCI,but increased on the 5th and 7th day after PCI.CTRP12 should be an independent influencing factor for the severity of coronary artery disease in the patients with AMI.

WS9326A is a peptide antibiotic containing a highly unusual N-methyl-E-2-3-dehydrotyrosine (NMet-Dht) residue that is incorporated during peptide assembly on a non-ribosomal peptide synthetase (NRPS). The cytochrome P450 encoded by sas16 (P450_Sas) has been shown to be essential for the formation of the alkene moiety in NMet-Dht, but the timing and mechanism of the P450_Sas-mediated α,β-dehydrogenation of Dht remained unclear. Here, we show that the substrate of P450_Sas is the NRPS-associated peptidyl carrier protein (PCP)-bound dipeptide intermediate (Z)-2-pent-1'-enyl-cinnamoyl-Thr-N-Me-Tyr. We demonstrate that P450_Sas-mediated incorporation of the double bond follows N-methylation of the Tyr by the N-methyl transferase domain found within the NRPS, and further that P450_Sas appears to be specific for substrates containing the (Z)-2-pent-1'-enyl-cinnamoyl group. A crystal structure of P450_Sas reveals differences between P450_Sas and other P450s involved in the modification of NRPS-associated substrates, including the substitution of the canonical active site alcohol residue with a phenylalanine (F250), which in turn is critical to P450_Sas activity and WS9326A biosynthesis. Together, our results suggest that P450_Sas catalyses the direct dehydrogenation of the NRPS-bound dipeptide substrate, thus expanding the repertoire of P450 enzymes that can be used to produce biologically active peptides.

Objective To analyze the serological markers and surgical indicators associated with biliary complications after orthotopic liver transplantation, explore their influencing factors and predictive indicators. Methods A retrospective analysis was performed for the clinical data of 101 patients who underwent orthotopic liver transplantation in Renmin Hospital of Wuhan University from January 2016 to June 2022, according to the presence or absence of biliary complication (BC) at 6 months after surgery, they were divided into BC group with 21 patients and non-BC group with 80 patients.The t -test or the Mann-Whitney U test was used for comparison of continuous data between groups, and the chi-square test was used for comparison of categorical data between groups.Univariate and multivariate Logistic regression analyses were performed, and the receiver operating characteristic (ROC) curve was used to evaluate the predictive performance of combined indicators. Results Among the 101 patients, 21(20.8%) experienced BC.The multivariate Logistic regression analysis showed that MELD score (odds ratio[ OR ]=0.134, 95% confidence interval[ CI ]: 0.031-0.590, P =0.008), SⅡ/Alb ( OR =1.415, 95% CI : 1.181-1.696, P =0.001), and plasma transfusion volume ( OR =1.001, 95% CI : 1.000-1.002, P =0.032) were independent risk factors for the development of BC in patients after liver transplantation.MELD score, SⅡ/Alb, plasma transfusion volume, MELD+SⅡ/Alb, and MELD+SⅡ/Alb+plasma transfusion volume had an area under the ROC curve of 0.712, 0.870, 0.712, 0.900, and 0.918, respectively, in predicting BC after liver transplantation. Conclusion SⅡ/Alb, plasma transfusion volume and MELD score are independent risk fators for BC after liver transplantation.The combination of three indicators has good predictive value and clinical guiding significance for BC after liver transplantation.