OBJECTIVES: To investigate the effect of BRD4 inhibition on migration of esophageal squamous cell carcinoma (ESCC) cells with low acetyl-CoA carboxylase 1 (ACC1) expression. METHODS: ESCC cell lines with lentivirus-mediated ACC1 knockdown or transfected with a negative control sequence (shNC) were treated with DMSO, JQ1 (a BRD4 inhibitor), co-transfection with shNC-siBRD4 or siNC with additional DMSO or C646 (an ahistone acetyltransferase inhibitor) treatment, or JQ1combined with 3-MA (an autophagy inhibitor). BRD4 mRNA expression in the cells was detected using RT-qPCR. The changes in cell proliferation, migration, autophagy, and epithelial-mesenchymal transition (EMT) were examined with CCK8 assay, Transwell migration assay, and Western blotting. RESULTS: ACC1 knockdown did not significantly affect BRD4 expression in the cells but obviously increased their sensitivity to JQ1. JQ1 treatment at 1 and 2 μmol/L significantly inhibited ESCC cell proliferation, while JQ1 at 0.2 and 2 μmol/L promoted cell migration. The cells with ACC1 knockdown and JQ1 treatment showed increased expresisons of vimentin and Slug and decreased expression of E-cadherin. BRD4 knockdown promoted migration of ESCC cells, and co-transfection with shACC1 and siBRD4 resulted in increased vimentin and Slug expressions and decreased E-cadherin expression in the cells. C646 treatment of the co-transfected cells reduced acetylation levels, decreased vimentin and Slug expressions, and increased E-cadherin expression. Treatment with JQ1 alone obviously increased LC3A/B-II levels in the cells either with or without ACC1 knockdown. In the cells with ACC1 knockdown and JQ1 treatment, additional 3-MA treatment significantly decreased the expressions of vimentin, Slug and LC3A/B-II and increased the expression of E-cadherin. CONCLUSIONS BRD4 inhibition promotes autophagy of ESCC cells via a histone acetylation-dependent mechanism, thereby enhancing EMT and ultimately increasing cell migration driven by ACC1 deficiency.
Humans ; Cell Movement ; Transcription Factors/metabolism* ; Esophageal Neoplasms/metabolism* ; Cell Line, Tumor ; Cell Cycle Proteins ; Azepines/pharmacology* ; Epithelial-Mesenchymal Transition ; Carcinoma, Squamous Cell/metabolism* ; Esophageal Squamous Cell Carcinoma ; Triazoles/pharmacology* ; Nuclear Proteins/genetics* ; Cell Proliferation ; Acetyl-CoA Carboxylase/genetics* ; Transfection ; Autophagy ; Bromodomain Containing Proteins

Objective:To investigate the role of iron death in paraquat (PQ) -induced alveolar epithelial mesangialization (EMT) .Methods:In August 2023, the appropriate PQ staining concentration as well as the intervention concentration of lipoinhibitor-1 (Lip-1) were screened by CCK8 method. The RLE-6TN cells were divided into three groups, which were control group, PQ group and iron death inhibition group, 200 μmol/L PQ solution was given to the PQ group, and PQ 200 μmol/L and 0.1 μmol/L Lip-1 solution was given to the iron death inhibition group, the control group was added the same amount of cell medium. morphological changes and migratory viability of the cells in each group were observed at 12 h, 24 h and 48 h after the poisoning, and the contents of ferrous ions (Fe 2+), reactive oxygen radicals (ROS), glutathione (GSH), superoxide dismutase (SOD), and malondialdehyde (MDA) were detected in each group; meanwhile, qRT-PCR and western-blot were used to determine the molecular expression of E-cadherin, α-smooth muscle actin (α-SMA), and Collagen I in the cells in each group. The difference between group was compared by ANOVA, and the further pairwise comparison was conducted by Bonferroni method. Results:Cell viability was detected using CCK8, and the results showed that the cell survival rate of RLE-6TN cells treated with 200 μmol/L PQ+0.1 μmol/L Lip-1 solution was 56.6%. The migration activity of RLE-6TN cells in the iron death inhibition group was weaker than that in the PQ group after 24 and 48 hours of exposure, and the degree of EMT changes in the cells was reduced compared to the PQ group. After 12, 24, and 48 hours of exposure, the Fe 2+ concentration, ROS fluorescence intensity, and MDA content in the iron death inhibition group decreased compared to the corresponding time points in the PQ group ( P<0.05/3), while compared with PQ group, the GSH concentration and SOD concentration increased compared to the corresponding time points in the PQ group ( P<0.05/3). The results showed that compared with the control group, the expression levels of E-cadherin mRNA and protein in PQ group and iron death inhibition group were both decreased ( P<0.05/3), while compared with PQ group, the expression levels of E-cadherin mRNA and protein in iron death inhibition group were increased ( P<0.05/3) ; Compared with the control group, the expression levels of α-SMA, Collagen I mRNA and protein in PQ group and iron death inhibition group cells increased ( P<0.05/3), while compared with PQ group, the expression levels of α-SMA, Collagen I mRNA and protein in iron death inhibition group cells decreased ( P<0.05/3) . Conclusion:Ferroptosis is involved in the EMT process of alveolar epithelial cells induced by PQ. Inhibiting ferroptosis can reduce cellular oxidative damage and alleviate the degree of cellular EMT.

Objective:To establish a rapid detection method based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS)for drug resistance of Carbapenem-resistant Enterobacteriaceae(CRE),so as to provide basis for enhancing clinically diagnostic effect.Methods:A total of 120 suspected CRE strains that were isolated from the clinical laboratory of Jiading District Central Hospital Affiliated to Shanghai University of Medicine&Health Sciences during October 2023 and September 2024 were collected,and the experimental detection of 44 strains was finally completed.The 44 suspected CRE bacterial strains were all from hospitalized patients.A rapid detection protocol based on the MALDI-TOF MS platform was initially established and improved,and experimental detection was carried out using the fully automatic rapid microbial mass spectrometry detection system(VITEK MS).The results of identification review and antimicrobial susceptibility test(AST)for VitEK-2 Compact strain was used as the reference standard.The screening results and detection efficacy of two kinds of detection methods were compared.Results:In the 44 suspected CRE strains,31 CRE strains were screened out by MALDI-TOF MS detection,and 31 CRE stains were screened out by VITEK-2 Compact recheck detection.In them,one strain of Enterobacter cloacae was confirmed as negativity by the MALDI-TOF MS test,but it was confirmed as CRE by the VITEK-2 Compact test.One strain of Klebsiella pneumoniae was confirmed as positivity by the MALDI-TOF MS test,but it was confirmed as mediator for carbapenem drugs by the VITEK-2 Compact test.The tested results of VITEK-2 Compact was used as standard,the results of four-grid table(2×2 contingency table)statistical method indicated that the sensitivity,specificity,accuracy rate and Youden index(R)of MALDI-TOF MS were respectively 96.7%,92.3%,100%and 89.1%.Conclusion:The rapid detection method based on the MALDI-TOF MS platform for CRE drug resistance has simple operation process and high accuracy rate,which can provide a basis for the application of targeted antibacterial drugs at early stage.

In-depth study of the components of polymyxins is the key to controlling the quality of this class of antibiotics.Similarities and variations of components present significant analytical challenges.A two-dimensional(2D)liquid chromatography-mass spectrometry(LC-MS)method was established for screening and comprehensive profiling of compositions of the antibiotic colistimethate sodium(CMS).A high concentration of phosphate buffer mobile phase was used in the first-dimensional LC system to get the components well separated.For efficient and high-accuracy screening of CMS,a targeted method based on a self-constructed high resolution(HR)mass spectrum database of CMS components was established.The database was built based on the commercial MassHunter Personal Compound Database and Library(PCDL)software and its accuracy of the compound matching result was verified with six known components before being applied to genuine sample screening.On this basis,the unknown peaks in the CMS chromatograms were deduced and assigned.The molecular formula,group composition,and origins of a total of 99 compounds,of which the combined area percentage accounted for more than 95％of CMS components,were deduced by this 2D-LC-MS method combined with the MassHunter PCDL.This profiling method was highly efficient and could distinguish hundreds of components within 3 h,providing reliable results for quality control of this kind of complex drugs.

Cancer is a serious global health problem and a major cause of human death. Conventional cancer treatments often run the risk of impairing vital organ functions. Anticancer peptides (ACPs) are considered to be one of the most promising therapeutic agents against common human cancers due to their small sizes, high specificity, and low toxicity. Since ACP recognition is highly limited to the laboratory, expensive, and time-consuming, we proposed pLM4ACP, a model for predicting ACPs based on machine learning and protein language models. In this model, the protein language model ProtT5 was used to extract the features of ACPs, and the extracted features were input into the support vector machine (SVM) classification algorithm for optimization and performance evaluation. The model showcased significantly higher accuracy than other methods, with the overall accuracy of 0.763, F1-score of 0.767, Matthews correlation coefficient of 0.527, and area under the curve of 0.827 on the independent test set. This study constructs an efficient anticancer peptide prediction model based on protein language models, further advancing the application of artificial intelligence in the biomedical field and promoting the development of precision medicine and computational biology.
Machine Learning ; Antineoplastic Agents/chemistry* ; Humans ; Peptides/chemistry* ; Support Vector Machine ; Algorithms ; Computational Biology/methods* ; Neoplasms/drug therapy*

In-depth study of the components of polymyxins is the key to controlling the quality of this class of antibiotics. Similarities and variations of components present significant analytical challenges. A two-dimensional (2D) liquid chromatography-mass spectrometr (LC-MS) method was established for screening and comprehensive profiling of compositions of the antibiotic colistimethate sodium (CMS). A high concentration of phosphate buffer mobile phase was used in the first-dimensional LC system to get the components well separated. For efficient and high-accuracy screening of CMS, a targeted method based on a self-constructed high resolution (HR) mass spectrum database of CMS components was established. The database was built based on the commercial MassHunter Personal Compound Database and Library (PCDL) software and its accuracy of the compound matching result was verified with six known components before being applied to genuine sample screening. On this basis, the unknown peaks in the CMS chromatograms were deduced and assigned. The molecular formula, group composition, and origins of a total of 99 compounds, of which the combined area percentage accounted for more than 95% of CMS components, were deduced by this 2D-LC-MS method combined with the MassHunter PCDL. This profiling method was highly efficient and could distinguish hundreds of components within 3 h, providing reliable results for quality control of this kind of complex drugs.

Objective:To quantitatively summarize the catechol-O-methyltransferase ( COMT) gene Val158Met polymorphism and the risk of obsessive-compulsive disorder (OCD). Methods:We searched databases including PubMed, Embase, Weipu and Wanfang for randomized controlled trials (RCTs) investigating the association between COMT gene polymorphisms and OCD up to November 1, 2023. Studies that reported genotype frequencies for both OCD patients and general healthy controls were included. Stata11 software was used to calculate pooled odds ratios ( OR) with 95% CI, perform heterogeneity test, and assess publication bias. Results:19 studies with 2, 393 OCD patients and 4, 134 healthy controls were included. The overall results showed that the Val158Met polymorphism was associated with OCD patients (allele model: OR=1.10, 95% CI: 1.02-1.20, P=0.016; homozygote model: OR=1.25, 95% CI:1.05-1.49, P=0.014; recessive model: OR=1.18, 95% CI:1.01-1.37, P=0.040). In the ethnic-stratified analysis, this significant association was mainly observed in Caucasians (allele model: OR=1.17, 95% CI:1.06-1.30, P=0.003; homozygote model: OR=1.35, 95% CI: 1.08-1.67, P=0.008; recessive model: OR=1.21, 95% CI: 1.01-1.44, P=0.041; dominant: OR=1.20, 95% CI: 1.01-1.43 P=0.040), but not in Asians. In gender-stratified analysis, Met-homozygote was associated with male OCD ( OR=1.75, 95% CI: 1.00-3.04, P=0.049). Moreover, the additional analysis found that the risk of OCD was significantly increased in Caucasian males (allele model: OR=1.48, 95% CI: 1.08-2.03, P=0.014; heterozygote model: OR=1.41, 95% CI: 1.03-1.93, P=0.030; dominant model: OR=1.60, 95% CI:1.08-2.38, P=0.020). Conclusion:This meta-analysis suggests that the COMT gene Val158Met polymorphism is associated with an increased risk of OCD in males, particularly in Caucasian males.

Objective:To survey the relationship among parents'rearing style,family conformity and obsess-ive-compulsive symptoms in patients with obsessive-compulsive disorder(OCD).Methods:Totally 216 patients di-agnosed with OCD were recruited(59 cases of mild obsessive-compulsive symptoms,121 cases of moderate obsess-ive-compulsive symptoms,36 cases of severe obsessive-compulsive symptoms).They were assessed with the Yale-Brown Obsessive-Compulsive Disorder Scale(Y-BOCS),Egma Minnen av Bardndosna Uppforstran(EMBU),and Family Accommodation Scale for Obsessive-Compulsive Disorder-Patient Version(FAS-PV).Results:The severe obsessive-compulsive symptom group had the lowest positive parenting style scores.Conversely,they had the high-est negative parental rearing scores and total FAS-PV scores(Ps＜0.01).The ordered logistic regression results showed that male patients had less severe obsessive-compulsive symptoms(OR=0.22).Patients with lower years of education(OR=0.87)and positive parental rearing scores(OR=0.98,0.97)had more severe obsessive-com-pulsive symptoms.Conversely,higher negative paternal rearing scores(OR=1.02)and total FAS-PV scores(OR=1.04)were associated with more severe obsessive-compulsive symptoms.Conclusion:This study suggests that the severity of obsessive-compulsive symptoms is related to parental rearing style and family accommodation.Patients with severe obsessive-compulsive symptoms are more likely to feel negative paternal rearing style and higher level of family compliance,and less likely to feel positive parental rearing style.

Objective:To survey the relationship among parents'rearing style,family conformity and obsess-ive-compulsive symptoms in patients with obsessive-compulsive disorder(OCD).Methods:Totally 216 patients di-agnosed with OCD were recruited(59 cases of mild obsessive-compulsive symptoms,121 cases of moderate obsess-ive-compulsive symptoms,36 cases of severe obsessive-compulsive symptoms).They were assessed with the Yale-Brown Obsessive-Compulsive Disorder Scale(Y-BOCS),Egma Minnen av Bardndosna Uppforstran(EMBU),and Family Accommodation Scale for Obsessive-Compulsive Disorder-Patient Version(FAS-PV).Results:The severe obsessive-compulsive symptom group had the lowest positive parenting style scores.Conversely,they had the high-est negative parental rearing scores and total FAS-PV scores(Ps＜0.01).The ordered logistic regression results showed that male patients had less severe obsessive-compulsive symptoms(OR=0.22).Patients with lower years of education(OR=0.87)and positive parental rearing scores(OR=0.98,0.97)had more severe obsessive-com-pulsive symptoms.Conversely,higher negative paternal rearing scores(OR=1.02)and total FAS-PV scores(OR=1.04)were associated with more severe obsessive-compulsive symptoms.Conclusion:This study suggests that the severity of obsessive-compulsive symptoms is related to parental rearing style and family accommodation.Patients with severe obsessive-compulsive symptoms are more likely to feel negative paternal rearing style and higher level of family compliance,and less likely to feel positive parental rearing style.

Objective:To establish a rapid detection method based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS)for drug resistance of Carbapenem-resistant Enterobacteriaceae(CRE),so as to provide basis for enhancing clinically diagnostic effect.Methods:A total of 120 suspected CRE strains that were isolated from the clinical laboratory of Jiading District Central Hospital Affiliated to Shanghai University of Medicine&Health Sciences during October 2023 and September 2024 were collected,and the experimental detection of 44 strains was finally completed.The 44 suspected CRE bacterial strains were all from hospitalized patients.A rapid detection protocol based on the MALDI-TOF MS platform was initially established and improved,and experimental detection was carried out using the fully automatic rapid microbial mass spectrometry detection system(VITEK MS).The results of identification review and antimicrobial susceptibility test(AST)for VitEK-2 Compact strain was used as the reference standard.The screening results and detection efficacy of two kinds of detection methods were compared.Results:In the 44 suspected CRE strains,31 CRE strains were screened out by MALDI-TOF MS detection,and 31 CRE stains were screened out by VITEK-2 Compact recheck detection.In them,one strain of Enterobacter cloacae was confirmed as negativity by the MALDI-TOF MS test,but it was confirmed as CRE by the VITEK-2 Compact test.One strain of Klebsiella pneumoniae was confirmed as positivity by the MALDI-TOF MS test,but it was confirmed as mediator for carbapenem drugs by the VITEK-2 Compact test.The tested results of VITEK-2 Compact was used as standard,the results of four-grid table(2×2 contingency table)statistical method indicated that the sensitivity,specificity,accuracy rate and Youden index(R)of MALDI-TOF MS were respectively 96.7%,92.3%,100%and 89.1%.Conclusion:The rapid detection method based on the MALDI-TOF MS platform for CRE drug resistance has simple operation process and high accuracy rate,which can provide a basis for the application of targeted antibacterial drugs at early stage.