OBJECTIVE: To explore changes of oxidation levels in lung tissues among the phosgeneexposedrats,and to investigate if oxidative stress would regulate autophagy to alleviate phosgene-induced acute lung injury (ALI) in rats. METHODS: Fifty rats were divided into five groups:control and treatment groups with phosgene treatment for 30 minutes and evaluated6,12,24 and 48 h later. The rats in the control group inhaled air through their noses, and the rats in the phosgene exposure group inhaled 41 mg/m³ of phosgene through their noses. After detecting relevant indicators,the 12 h post-exposure group was selected as the animal model for this study (hereinafter referred to as the phosgene group). Another 40 rats were divided into a control group (intraperitoneal injection of 0.9% normal saline),a phosgene group (41 mg/m³ of phosgene exposure for 30 min),and an RPM treatment group (intraperitoneal injection of 5 mg/kg of RPM after exposure) and the RPM control group (intraperitoneal injection of 5 mg/kg of RPM). Another 40 rats were divided into the control group (intraperitoneal injection of 0.9% normal saline), the phosgene group (41 mg/m³ of phosgene exposure for 30 min),and the NAC treatment group (the intraperitoneal injection of 200 mg/kg of NAC after exposure) and NAC control group (intraperitoneal injection of 200 mg/kg of NAC). These rats were observed for pathological changes in lung tissues; lung functions, lung moisture mass, lung coefficient, total protein concentration of bronchoalveolar lavage fluid (BALF), rat lung tissue ROS levels, lung tissue lipid peroxidation levels, anti-Oxidase (SOD, CAT) activity and GSH content, and redox and autophagy-related molecule mRNA and protein expression levels. RESULTS: Compared with the control group,histopathological observation at different time points after phosgene exposure showed alveolar fusion and increased inflammatory cells. Lung function decreased significantly (P<0.05 or 0.01). The lung moisture mass, lung coefficient and BALF total protein concentration were significantly increased (P<0.05 or 0.01). The levels of ROS and lipid peroxidation in lung tissues of rats were increased (P<0.01). The activity and expression levels of a series of antioxidant enzymes were changed (P<0.05 or 0.01),and the changes were most significant at 12 h after exposure. Compared with the control group, expression levels of autophagy-associated proteins, Beclin-1 and LC3II/LC3I, decreased in phosgene group,while the expression levels of P62 increased (P<0.05 or 0.01). Compared with phosgene group, the autophagy levels in RPM and NAC groups were up-regulated (P<0.05) and the phosgene induced ALI was alleviated. At the same time, the ROS level of lung tissue was decreased in the NAC treatment group. CONCLUSION: Oxidation levels of lung tissues increased at different time points after phosgene exposure. At the same time, oxidative stress regulated the level of autophagy. Administration of NAC alleviated oxidative stress,thereby upregulated autophagy levels and improved phosgene-induced ALI in rats.

OBJECTIVE: To study the role of the c-Jun N-terminal kinase (JNK) pathway-mediated ferroptosis in the proliferation and apoptosis of triple negative breast cancer (TNBC). METHODS: From January 2020 to May 2022,105 cases of TNBC tissues and adjacent tissues which were resected surgically in the First Affiliated Hospital of Jiamusi University were collected, TNBC cell line MDA-MB-231 and non TNBC cell lines MCF-7, SK-BR-3 were cultured. Expression levels of p-JNK and ferroptosis marker gene transferrin receptor 1 (TFR1) and glutathione peroxidase 4 (GPX4) in the tissues and the TNBC cells and non TNBC cells were detected. MDA-MB-231 cells in cultures were divided into the control and the JNK agonist groups (5 μmol/L ANISO),the ferroptosis inhibitor (2 μmol/L ferrostatin-1),and the agonist+inhibitor groups (5 μmol/L ANISO+2 μmol/L ferrostatin-1). After 24h treatment in each group, proliferation inhibition rates were detected using the CCK-8 assay,apoptosis rates by the TUNEL assay,activities of glutathione peroxidase (GSH),superoxide dismutase (SOD) and the content of malondialdehyde (MDA) were by kits,and expression levels of p-JNK, TFR1 and GPX4 were by PCR and western blot. RESULTS: Compared with adjacent tissues,expression levels of p-JNK and TFR1 in the TNBC tissues were decreased,but the levels for GPX4 were increased (P<0.05). p-JNK expressions were positively correlated with TFR1(correlation coefficient r=0.515,P<0.05) and negatively correlated with GPX4 (correlation coefficient r=-0.442,P<0.05). Compared with the MCF-7 and SK-BR-3 cells,expression levels of p-JNK and TFR1 in MDA-MB-231 were decreased,and expression levels of GPX4 were increased (P<0.05). Compared with the control group,proliferation inhibition rates,apoptosis rates,the expression levels of TFR4,and contents of MDA were increased (P<0.05),while expression levels of GPX4,and activities of GSH and SOD were decreased in the JNK agonist group (P<0.05). In addition,proliferation inhibition rates,apoptosis rates,activities of GSH and SOD,and contents of MDA showed no significant changes (P>0.05),while expression levels of TFR4 were decreased and expression levels of GPX4 were increased (P<0.05) in the ferroptosis inhibitor group. Compared with JNK agonist group, proliferation inhibition rates,apoptosis rates,expression levels of TFR4,contents of MDA were decreased (P<0.05), while expression levels of GPX4, and activities of GSH and SOD were increased in the agonist + inhibitor group (P<0.05). CONCLUSION: Activation of the JNK pathway inhibited proliferation and promoted apoptosis of TNBC cell by activating ferroptosis.

OBJECTIVE:To investigate expression and biological function of the transforming growth factor β-induced protein (TGFBI) in glioma. METHODS: The transcriptome data and the corresponding clinical information of tumor samples from glioma patients in the Cancer Genome Atlas (TCGA)、Chinese Glioma Genome Atlas (CGGA) and Rembrandt database were obtained to analyze potential associations between grades,subtypes, overall survival of glioma patients and TGFBI mRNA expression levels. Expression of TGFBI protein in 35 glioblastoma tissues and 5 normal brain tissues was analyzed using immunohistochemical method. Furthermore, the U87 and U373 cell lines were transfected with the constructed lentivirus vector containing shTGFBI target sequence to establish glioblastoma cell lines with low TGFBI expression and then to explore the effect of TGFBI on tumor proliferation、invasion、apoptosis and cell cycle. Finally,Gene set enrichment assay (GSEA) was used to predict the related pathways that TGFBI might participate in the occurrence and development of glioma. RESULTS: Results from TCGA、CGGA and Rembrandt databases showed that TGFBI expressions were positively correlated with the WHO grades (P<0.05). In addition,high expressions of TGFBI were correlated with inferior survival in glioma patients. Expressions of TGFBI in glioma tissues were significantly higher than that in normal brain tissues (P<0.05). The knockdown of TGFBI significantly inhibited proliferation and invasion of cancer cells、promoted cell apoptosis and promoted cell cycle progression from the G1 to the S phases. GSEA showed that high expressions of TGFBI were involved in activation of the TOLL-like receptors,NOD-like receptors and JAK-STAT signaling pathways. CONCLUSION: Overexpressions of TGFBI in gliomas promoted proliferation and invasion of tumor cells,inhibited cell apoptosis and induced cell cycle rearrangement. Thus,TGFBI might play an oncogene role in the occurrence and development of gliomas.

OBJECTIVE: Virtual screening of potential small molecule inhibitors of acetylcholinesterase (AChE) in Cistanche tubulosa compound preparations,while providing reference for the discovery of new AChE inhibitors. METHODS: A virtual screening database of Cistanche tubulosa, ginseng and astragalus was established based on the traditional Chinese medicine system pharmacology technology platform (TCMSP) and literature retrieval. The Glide molecule was docked by Rosetta software,and a wide range of Box was formed by using the amino acid residues around the active site of AChE receptor. Different types of components were used as probes to scan, and the lattice energy was calculated for energy ranking. After that, conformation of the ligand in the range of Box was searched,and the results were scored according to the different conformation, direction,position and energy of the ligand. Finally,the SPR technology was used to verify the binding force of the main active components and AChE protein. RESULTS: The binding mode and binding energy of AChE protein to the typical lead compounds selected from each system were good. In the molecular docking system of all compounds, Cistanche tubulosa energy scored between -9.397~-0.858 kcal/mol, ginseng energy score between -9.929~-0.211 kcal/mol and astragalus energy scored between -9.408~-1.555 kcal/mol. The docking energies of all compounds were less than -7.0 kcal/mol. The results of SPR affinity test showed that quercetin, kaempferol and genistein all showed good binding ability to AChE protein. CONCLUSION: The presence of AChE inhibitors in Cistanche tubulosa compound preparation was screened based on molecular docking technique, and the good affinity between AChE protein and active components was verified by SPR technique.

OBJECTIVE:To investigate expression and diagnostic values of LncRNA CUST_49525 and CUST_47163 in peripheral blood lymphocytes in patients with acute myeloid leukemia (AML). METHODS: Real time fluorescence quantitative PCR (qPCR) was used to detect expression levels of LncRNA CUST_49525 and CUST_47163 in peripheral blood lymphocytes of patients and healthy controls. The cytokinesis-arrest micronucleus test was used to detect micronuclei levels in lymphocytes. The collected data were evaluated using the non-parametric Mann Whitney U test to determine significant differences and ROC curves were drawn to determine diagnostic significance. RESULTS: Compared with the control group,the expression levels of both LncRNA CUST_49525 and CUST_47163 from the patients were significantly down-regulated (P<0.01), while the micronucleus rates were significantly increased (P<0.01). The ROC curve analyses show that the area under the ROC curve of LncRNA CUST_49525 in the patients was 0.742,with a 95%CI of 0.627-0.858;and that for LncRNA CUST_47163 was 0.859,with a 95%CI of 0.769-0.949. The area under the ROC curve for the micronucleus rates was 0.948,with a 95%CI of 0.899-0.997 (P<0.01). The area under the ROC curve for the combined diagnosis of the three was 0.970,with a 95%CI of 0.931-1.000 (P<0.01). CONCLUSION: The expression levels of LncRNA CUST_49525 and CUST_47163 in peripheral blood lymphocytes of AML patients were abnormally down-regulated. The combined diagnostic effect of the two together with the micronucleus rate was superior to a single indicator,and can be a potential reference indicator for AML diagnosis.

OBJECTIVE:This study aimed at detecting whether microplastics existed in human tophi; and then qualitatively and quantitatively analyzing the detected microplastics; identifying compositions of tophi,exploring possible roles of microplastic accumulation in tophi formation in the joints,and then providing new ideas for the treatment of gout. METHODS: Samples of gouty tophus from 5 male patients (serum uric acid>500 μmol/L) were included for analysis,and pyrolysis-gas chromatography-mass spectrometry (Py-GC/MS) was used to detect 10 types of microplastics that might exist in tophi. RESULTS: More than or equal to 1 type of microplastic was detected in the tophi of 4 patients,and a total of 4 different types of microplastics (polystyrene,polyethylene,polyvinyl chloride,polyadiohexylenediamine) were identified. The results demonstrate that polystyrene (PS) and polyethylene (PE) had the highest detection rate,with average contents of 20.300 and 115.157 mg/kg,respectively. CONCLUSION: Microplastics can be detected in human tophi,which may be involved in and may play a certain role in the formation of human tophi.

OBJECTIVE: Tongue squamous cell carcinoma (TSCC) had a high rate of metastasis and postoperative recurrence. Our study objective was to use a nomogram model to provide prognosis of TSCC patients undergoing surgery. METHODS: From 2008 to 2019,a retrospective analysis was performed on 167 patients with TSCC who were treated in the Cancer Hospital Affiliated to Shantou University Medical College. Cox regression analysis was used to identify independent prognostic factors associated with the overall survival (OS) which was used to build a nomogram and to predict OS. The predictive accuracy of the model was evaluated by calibration curve, decision curve (DCA) and the concordance index (C-index). RESULTS: Results from the multivariate Cox regression and Kaplan-Meier survival analyses show that B-factor (BF), monocyte count,lymphocyte-monocyte ratio,and tumor lymph node metastasis (TNM) stage were independent prognostic factors,which were used to build the prognostic nomogram model. The C-index of the nomogram was 0.754[95%CI(0.672,0.835)],which was higher than that of the TNM stage 0.679[95%CI(0.602,0.755)]. The Akaike information criterion (AIC) and Bayesian information criterion (BIC) of the nomogram were 312.134 and 318.239,respectively. Both were lower than 319.163 and 320.689 for TNM stages alone,indicating that the nomogram has better goodness of fit for OS prediction. The calibration curves of the nomogram show good consistency between the predicted OS probabilities and the actual OS value of patients. In addition, timedependent C-index analysis and DCA results show that the prognostic nomogram model had good predictive accuracy and discriminability compared with TNM stage. CONCLUSION: A nomogram model based on clinicopathological features and preoperative inflammation-related indicators may be useful in predicting the prognosis of TSCC patients undergoing surgery.

OBJECTIVE:To investigate expression and clinical significance of aberrantly expressed genes in esophageal cancers (ESCA) by mining them from the Gene Expression Omnibus (GEO) database through bioinformatics technology. METHODS: The ESCA microarray datasets GSE38129 and GSE20347 were downloaded from the GEO database using the GEOquery package in R. After the batch effect was removed from the merged dataset by the sva package,the merged dataset was screened for differentially expressed genes (DEGs) using the Limma package, and gene ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed on the DEGs using the clusterProfiler package. Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed on the DEGs at the STRING website,and protein interaction network analysis (PPI) was performed on the DEGs,and core modules and core genes (hub genes) were extracted by using MCODE and CytoHubba plugins. The Hub gene was entered into the University of Alabama at Birmingham Cancer Database (UALCAN) to analyze the relationship between its expression level and esophageal cancer stage,methylation level, and TP53 mutation, etc. Finally, the core genes were verified with the help of microarray data GSE70409. RESULTS: 390 DEGs were screened in the normalized merged dataset:166 up-regulated DEGs and 224 down-regulated DEGs.GO analysis showed that they were mainly involved in biological processes such as mitotic cell cycle phase transition,extracellular matrix production,and epidermal development. KEGG enrichment analysis showed that DEGs were associated with signaling pathways such as cell cycle, extracellular matrix receptor interactions, and amoebiasis. The obtained PPIs were imported into Cytotec. The obtained PPIs were input into Cytoscape software,and a total of 20 key genes were screened. The key genes were inputted into the UALCAN database for analysis,and the mRNA expression levels of three genes (CDK1,TOP2A,AURKA) were screened out to be significantly higher in esophageal cancer than in normal tissues (P<0.05),and the expression rates of these three genes were higher in cases with clinical stage of esophageal cancer,mutation of TP53,and methylation of the promoter of this gene,and the differences were all statistically significant (P<0.05). The expression rates of these three genes were significantly higher in male patients than in female patients (P<0.05),and the expression rates of these three genes were significantly higher in patients whose ages were concentrated in the 41-60 range than in other age groups (P<0.05). By Kaplan-Meier survival curve analysis of Gene Expression Profiling Interactive Analysis (GEPIA) database,the overall survival of ESCA patients with high expression of CDK1 gene was shorter than that of those with low expression (P=0.036),and the overall survival of ESCA patients with high expression of AURKA gene was shorter than that of those with low expression (P=0.033), therefore CDK1, AURKA gene expression was negatively associated with the prognosis of esophageal cancer. CONCLUSION: Using a combination of bioinformatics techniques,three core genes were identified,the expression of which was higher in esophageal cancer than in normal tissues. Expression of CDK1,TOP2A,and AURKA was positively correlated with the stage of the tumor,the level of methylation of the promoter of the genes,and the mutation status of TP53. Among them,CDK1 and AURKA had the potential to become a new molecular marker for clinical diagnosis and prognosis of esophageal cancer.

OBJECTIVE:To analyze the prevalence,age distribution and annual infection trend of 17 high-risk HPV subtypes in Handan. METHODS: The high-risk HPV results of 64 317 female patients who came to Handan Central Hospital for cervical cancer screening from January 1,2014 to December 31,2021 were collected. The overall prevalence, distribution types, age-specific prevalence and annual trends were analyzed. RESULTS: Among 64 137 female patients screened for cervical cancer,16 096 were positive for high-risk HPV, with an overall high-risk infection rate of 25.1%, with 11 902(73.9%) of single type infection,and 4 194(26.1%) of double or more types of multiple infection. The top five subtypes of infection rate were HPV16,HPV52,HPV58,HPV53 and HPV51,accounting for 16.5%,14.5%,10.5%,7.9% and 7.2% of the positive items respectively. The highest infection rate of high-risk HPV was ≤ 20 years old,with the infection rate of 70.6%,followed by ≥ 60 years old,with the infection rate constituent ratio of 29.4%. The age group with the highest infection rate of single high-risk HPV was 41-50 years old, and the constituent ratio of infection rate was 77.8%. The age group with the highest infection rate of multiple severe high-risk HPV in each age group was ≤ 20 years old,and the constituent ratio of infection rate was 39.1%. The infection rate of single high-risk HPV and multiple severe high-risk HPV in each age group was statistically different (P<0.01). The annual high risk HPV infection rate showed a downward trend (P<0.01). CONCLUSION: The infection rate of high-risk HPV among women in Handan was high. The common subtypes of high-risk HPV infection were HPV16,HPV52,HPV58,HPV53,and HPV51.And the distribution was U-shaped with age.According to the analysis,the annual infection rate showed a downward trend from 2014 to 2021. The prevalent characteristics of HPV should help to guide HPV vaccinations and cervical cancer screenings.

OBJECTIVE:Based on flow cytometry,genotoxicity of biodegradable patch was investigated by studying the induction of micronuclei in peripheral reticulocytes (MN-RET) and mature erythrocytes (MN-NCE) in mice under different administration conditions. METHODS: For the short-term administration condition, mice were intraperitoneally injected with vehicle control,test solution,and cyclophosphamide solution (25,50 mg/kg) once a day for 3 days. Then,peripheral blood samples were collected,and the micronucleus were detected by flow cytometry. For the long-term administration condition, mice were continuous administrated for 14 days. Then,peripheral blood samples were collected,and spleens were enucleated. Micronucleus frequencies as well as splenic cell apoptosis were detected by flow cytometry. In addition, spleens were histopathologically examined after hematoxylin-eosin staining (HE). RESULTS: There was no significant differences in the percentage of MN-RET and MN-NCE in both treatment and control groups under the two different administration conditions (P> 0.05). In addition, there was no significant difference in the apoptosis rate of spleen cells between the two groups (P>0.05),and there were no significant pathological changes in the spleen tissue. On the other hand, significant and positive results were observed with the cyclophosphamide groups. CONCLUSION: The micronucleus test results show that the biodegradable patches had no potential genotoxic risk. In addition,the selection and analysis of MN-RET and MN-NCE in mouse peripheral blood under different dosing cycle conditions was useful in detecting genotoxic risk of chemicals.