OBJECTIVE： To investigate effects of the Yes-associated protein-transcriptional enhanced associate domain (YAP-TEAD) small-molecule inhibitor GNE-7883 on malignant phenotypes and molecular mechanisms in esophageal squamous cell carcinoma (ESCC) cells. METHODS： KYSE410 and KYSE510 cells were treated with varying concentrations of GNE-7883 (8，16，50，and 100 μmol/L) for 24 or 48 hours，with DMSO as control. Cell proliferation， colony formation， migration， and apoptosis were assessed. Molecular expression changes were analyzed by qPCR and Western blot. RESULTS： Compared to controls，50 and 100 μmol/L of GNE-7883 significantly inhibited proliferation and colony formation in both cell lines (P<0.01)，and induced apoptosis under serum-free conditions (P<0.05). Under serum-free conditions， 8 and 16 μmol/L of GNE-7883 significantly inhibited cell migration (P<0.01). Normal cultures treated with 50 and 100 μmol/L of GNE-7883 reduced CTGF，CYR61，c-Myc，p-ERK and p-AKT levels. Serum-free conditions with 50 and 100 μmol/L of GNE-7883 increased Cleaved PARP and Cleaved Caspase-3，while decreasd CTGF，CYR61， Bcl-2， Bcl-xL， c-Myc， p-ERK and p-AKT. Serum-free conditions with 8 and 16 μmol/L of GNE-7883 down-regulated CTGF， CYR61， MMP14， and p-ERK. CONCLUSION： GNE-7883 significantly suppressed proliferation， migration， and survival of esophageal squamous cell carcinoma (ESCC) cells and markedly inhibited activities of the MAPK/ERK and AKT/mTOR signaling pathways. It could be useful as a therapeutic candidate for ESCC.

OBJECTIVE： This study aimed to screen for methylation changes at MGMT CpG sites and their biological significance in blood lymphocytes from healthy adults who were exposed to short-term PM_2.5. METHODS： Thirty healthy male undergraduates from Hebei Medical University participated in a 35-day panel study， during which daily PM_2.5 exposure was assessed based on individual time-activity patterns. MGMT methylation was quantified by pyrophosphate sequencing of DNA extracted from peripheral blood lymphocytes at low and high PM_2.5 exposure time points. Additionally， urinary 1-hydroxypyrene (1-OHP)， 8-hydroxy- 2'-deoxyguanosine (8-OHdG)， and plasma tumor necrosis factor-alpha (TNF-α) levels were analyzed. RESULTS： The average ambient PM_2.5 concentration for the three days before the high exposure (110.09 μg/m³) was 2.32 times higher than at low exposure (47.37 μg/m³)，while urinary 1-OHP levels，indicating internal exposure，were 1.73 times higher (P<0.01). Urinary 8-OHdG and plasma TNF-α levels were 1.18-fold and 1.66-fold higher，respectively，than those under low exposure (all P<0.01). Increase in oxidative DNA damage and inflammatory response was accompanied by a decrease in the methylation levels of MGMT CpG sites 1， 3，4，5，6，and 10 by 37.31%，42.60%，19.69%，15.29%，34.95% and 32.12%，respectively (all P<0.05). Hypomethylation at CpG sites 1，3，and 10 was negatively correlated with urinary 1-OHP (β≥-0.40，all P<0.05)，8-OHdG (β≥-0.46，all P<0.05)，and TNF-α (β≥-0.26，all P<0.05). Altered methylation at these hot CpG sites was negatively correlated with the moving average concentrations of PM_2.5 (lag0-2) (β≥-0.53，all P< 0.05). CONCLUSION： MGMT hypomethylation modifications at hot CpG sites 1，3 and 10 were found to be associated with both external and internal PM_2.5 exposure as well as biological effects，indicating their potential as biomarkers for PM_2.5 exposure in the population.

OBJECTIVE： Cervical adenocarcinoma is markedly distinct from cervical squamous cell carcinoma due to its highly aggressive nature， lower response to radiotherapy， poor prognosis and reduced survival rates. This study aimed to investigate the expression level of RUNX1 in cervical adenocarcinoma tissues and its association with patient prognosis. METHODS： Tumor tissue samples from 37 cervical adenocarcinoma patients were collected. RUNX1 expression in tumor tissues was detected by immunohistochemistry (IHC)， and its correlation with clinicopathological parameters and prognosis was statistically analyzed. The Human Protein Atlas (HPA) database was used to analyze the protein expression level of RUNX1 in cervical adenocarcinoma tissues. Gene Set Variation Analysis (GSVA) scoring and the TIMER2.0 platform were utilized to perform signaling pathway enrichment and immune cell infiltration analyses， respectively， using cervical adenocarcinoma datasets from The Cancer Genome Atlas (TCGA) to preliminarily elucidate the mechanistic role of RUNX1 in cervical adenocarcinoma. RESULTS： Immunohistochemistry results demonstrated that RUNX1 expression in cervical adenocarcinoma showed significant differences correlated with patient prognosis and pathological stage (P<0.05). Patients in the RUNX1 high-expression group exhibited shorter overall survival and more advanced pathological stages. Analysis of the HPA database revealed that the protein expression level of RUNX1 was significantly higher than in normal cervical tissues. GSVA of cervical adenocarcinoma datasets from TCGA revealed that RUNX1 high expression was predominantly enriched in pathways including mitotic spindle assembly，Hedgehog signaling，TGF-β signaling，Wnt/β-catenin signaling， and p53 signaling. Immune cell infiltration analysis via TIMER2.0 indicated a significant correlation between RUNX1 expression and neutrophil infiltration. CONCLUSION： RUNX1 was found to be highly expressed in cervical adenocarcinoma and its overexpression was significantly associated with adverse prognosis，suggesting its potential as a prognostic biomarker for cervical adenocarcinoma patients.

OBJECTIVE： To use network pharmacology analysis together with molecular docking and in vitro cell experiments for evaluating therapeutic potential of ginseng cyclopentadienol (PND) on Alzheimer's disease，and for understanding its protective effect on neurons. METHODS： Targets of PND in the treatment of Alzheimer' s disease were screened based on PharmMapper， GeneCards， STRING and other databases， and a protein interaction network was constructed. Characteristic pathways were identified by GO and KEGG pathway enrichment analysis and then verified by molecular docking technology. An L-glutamate-induced cell injury model was established using human neuroblastoma SH-SY5Y cells. After treatment of these cells with different concentrations of PND (10，20，40 μmol/L)， cell activity was detected by CCK-8 method，and apoptosis was detected by flow cytometry. RESULTS： A PND-core target-characteristic pathway-Alzheimer's disease network was constructed， and the important targets regulated by PND were found to be ALB， AKT1， EGFR， SRC， CASP3， NFKB1， PTGS2， PPARG， etc. Multiple pathways were identified to involve EGFR， C-type lectin receptor signaling，RAS signaling，PI3K-Akt signaling and Alzheimer's disease signaling. The molecular docking results showed that PND had a good regulatory effect on targets such as CYP3A4，APP，and EGFR. Results from the in vitro cell experiments showed that the survival rate of SH-SY5Y cells in the L-glutamate-induced nerve injury model group was reduced，and the cell survival rate in the 20 μmol/L PND treatment group was higher than that in the model group (P<0.01)；the total apoptosis rate of nerve cells in the model group was 45.23%，and the apoptosis rates of cells in the 10，20 and 40 μmol/L PND treatment groups were 41.34%， 36.96% and 21.86% ， respectively， which were lower than those in the model group (P<0.05 or P<0.01). CONCLUSION： PND showed a neuroprotective effect which may be useful for treatment of Alzheimer's disease.

OBJECTIVE： To identify cofilin-1， a characteristic peripheral blood marker for recurrent metastasis of lung adenocarcinoma， by enrichment and mass spectrometry of plasma low-abundance proteins from lung adenocarcinoma patients using novel nanomaterials and to explore its relationship with clinicopathological features and prognosis of lung adenocarcinoma. METHODS： Twenty-four patients with complete clinical information of lung adenocarcinoma and 52 patients with benign lung disorders (17 tuberculosis，25 inflammatory granuloma and 10 malignant tumors) diagnosed by postoperative pathology were selected. Peripheral blood was collected from the patients，and low abundance proteins in the samples were enriched by kit nanomagnetic beads and subsequently identified by high resolution mass spectrometry. The target-decoy (TDfdr) algorithm was applied to screen differentially expressed genes in R software to control the pseudo-discovery rate within 0.05. RNA expression profiles and clinical data of 598 lung adenocarcinoma tissues were downloaded from The Cancer Genome Atlas (TCGA) database for Kaplan-Meier survival analysis and Log-rank test. RESULTS： A total of 6 563 proteins were identified by mass spectrometry， and 105 differentially expressed proteins were obtained after screening by the TDfdr algorithm (39 of which were lowly expressed and 66 were highly expressed in lung cancer plasma samples. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the low-expressed proteins were mainly involved in pathways related to the maintenance of the cell's material exchange，structural stability，and normal motor functions，which are involved in constituting the molecular basis of tissue development， homeostasis， and damage repair. On the other hand， the highly expressed proteins were mainly involved in pathways related to metabolic reprogramming，cell division，and motor migration，which help to maintain the survival and growth of cancer cells and promote the proliferation and metastasis of cancer cells. Bioinformatics analysis revealed that the actin depolymerization factor cofilin-1 was highly expressed in the plasma of lung adenocarcinoma patients (P<0.05)， and the high expression of cofilin-1 in TCGA lung adenocarcinoma tissues was negatively correlated with the overall survival of the patients (r=-1.48，P<0.05). CONCLUSION： There are large differences in plasma low abundance protein expression between patients with lung adenocarcinoma and those with benign lung diseases. High expression of cofilin-1 in cancer plasma and tissue correlated with poor prognosis of the patients，and it may therefore be useful as a plasma molecular marker for lung adenocarcinoma.

OBJECTIVE： To investigate the expression characteristics， prognostic significance and functional role of cytoplasmic polyadenylate-binding protein 4 (PABPC4) in liver cancers. METHODS： RNA sequencing data of liver cancers from The Cancer Genome Atlas (TCGA) were downloaded to analyze relationships between PABPC4 expression and patient prognosis. PABPC4 knockdown and overexpression liver cancer cell lines were constructed， and expression of PABPC4， its stemness-related indexes mRNA and protein in cells were detected by real-time quantitative PCR and Western blot. Effects of PABPC4 on cell proliferation，migration，invasion and stemness were studied by CCK-8 method，plate colony formation assay， cell scratch assay and Transwell assay. RESULTS： Expression levels of PABPC4 in liver cancer tissues gradually increased with increase of pathological grade. Patients with high expression had shorter overall survival and recurrence-free survival time (P<0.05). In tumor tissues with the high expression showed upregulated PABPC4，Wnt signaling pathway-related proteins. The successfully constructed HLF cells showed stable knockdown of PABPC4 and Huh7 cells with stable overexpression. Functional experiments showed that knockdown of PABPC4 showed significantly inhibited cell proliferation， migration， invasion and colony formation (P<0.05). Overexpression of PABPC4 had the opposite effect (P<0.05). In addition， after high expression of PABPC4， the mRNA expression levels of cell stemness-related indicators (Nanog， OCT4， CD133， SOX2) were significantly increased. CONCLUSION： PABPC4 was highly expressed and was correlated with poor prognosis in liver cancer patients. PABPC4 enhanced cancer cell stemness and promoted proliferation， survival， migration， and invasion of cancer cells. Therefore， PABPC4 is a potential target for prognostic prediction and therapeutic intervention in liver cancer.

OBJECTIVE： This study aimed to investigate effect of Danshen (Salvia miltiorrhiza) preparation on blood concentration of imatinib and N-desmethyl imatinib in rats. METHODS： Healthy male SD rats were randomly divided into three groups of 10 per group：control，Danshen tablet of 486 mg/kg and Danshen dripping pill of 72.9 mg/kg. All three groups were given imatinib at 30 mg/kg. Each group was given by intragastric administration for 14 consecutive days. Plasma samples were collected on day 1 and day 14. Blood concentrations of imatinib and N-desmethyl imatinib were determined by ultra-performance liquid chromatography-mass spectrometry (HPLC-MS/MS). Calculation of pharmacokinetic parameters of imatinib and N-desmethyl imatinib using DAS 2.0 software. RESULTS： On the 1st day after administration，the area under the concentration-time curve (AUC_0-∞) of imatinib in the compound Danshen tablet group was reduced by 19.00% (P<0.05) compared with the control group. On the 14th day after administration，the AUC_0-24 and peak concentration (Cmax) of imatinib in the compound Danshen tablet group were reduced by 19.45% and 17.84%， respectively (P<0.05). The AUC_0-24 and AUC_0-∞ of N-desmethylimatinib were reduced by 29.87% and 35.27%， respectively (P<0.05). On the 1st and 14th days after administration，there were no significant differences in the pharmacokinetic parameters of imatinib and N-desmethylimatinib between the compound Danshen dripping pill group and the control group (P>0.05). CONCLUSION： Compound Danshen tablets affected the plasma concentrations of imatinib in rats while compound Danshen dripping pills had no significant effect on the concentrations of imatinib and N-desmethylimatinib in rat plasma.

OBJECTIVE： To investigate the mechanisms by which malonyl-CoA decarboxylase (MLYCD) regulates platinum resistance in ovarian cancer and its clinical application value. METHODS： Immunohistochemical staining (IHC) was performed on ovarian cancer tissue microarrays to validate MLYCD expression levels in patients with varying platinum sensitivities. Using the Clinical Proteomic Tumor Analysis Consortium (CPTAC) and The Cancer Genome Atlas (TCGA) databases， protein and mRNA expression levels of MLYCD among these patients were evaluated and their association with clinical characteristics was examined. The pRRophetic algorithm was used to evaluate sensitivity of the samples to cisplatin. Proteins significantly correlated with MLYCD expression were selected to construct a protein-protein interaction (PPI) network，and the GEPIA2 database was used to retrieve MLYCD-related genes at the transcriptome level. Related signaling pathways were enriched using the KEGG and GO methods. The immune infiltration status of patients' tumor tissues was evaluated using the Estimate algorithm，tumor immune dysfunction and exclusion (TIDE) score，and Immunophenoscore (IPS). Western blot (WB) was used to validate the expression of MLYCD protein in cisplatin-sensitive/resistant ovarian cancer cell lines. RESULTS： The tissue microarray IHC results， together with CPTAC data， showed that MLYCD expression was higher in platinum-resistant patients than in platinum-sensitive patients and exhibited an increasing trend with tumor substage progression (P=0.046). Both CPTAC and TCGA data indicated that MLYCD expression was positively correlated with cisplatin resistance (r> 0.3， P<0.05). Furthermore， MLYCD and its interacting proteins were mainly involved in metabolic reprogramming that promoted lipid catabolism. The Estimate，TIDE，and IPS scores indicated that，compared to the low-expression group， ovarian tumors with high MLYCD expression harbored greater infiltration of dysfunctional immune cells and exhibited reduced antitumor activity. Western blot experiments further confirmed a significant increase in MLYCD expression in platinum-resistant ovarian cancer cell lines (P<0.05). CONCLUSION： MLYCD was highly expressed in platinum-resistant ovarian cancer tissues and participated in metabolic reprogramming toward lipid catabolism to supply energy for ovarian cancer cells. Thus，MLYCD may serve as a biomarker for platinum resistance and a potential therapeutic target in ovarian cancer.

OBJECTIVE： This study evaluated the concordance between immunocytochemistry(ICC) of PAX8，WT-1，p53 and p16 in effusion sample and immunohistochemistry (IHC) of primary ovarian tumors； assessed the diagnostic ability of the markers for diagnosing high-grade serous ovarian cancer(HGSOC)； and developed specific criteria for effusion cytology. METHODS： This study included 128 patients with effusion cytology diagnosed as adenocarcinoma and histopathologically confirmed for stage III-IV HGSOC. PAX8，WT-1 positivity，and p53 mutation status were conducted as diagnostic criteria for HGSOC. The Kappa test assessed the concordance between PAX8， WT-1， p53， and p16 expression in serous effusion samples and primary ovarian tumors. The number of definitively classified cases was recorded. RESULTS： The concordance rate of PAX8 expression between effusion and primary sites was 98.31%，with a Kappa value of 0.944 (P<0.05). For WT-1，the concordance rate was 84.43% and the Kappa value was 0.783 (P<0.05). The concordance rates for p53 and p16 were 71.9% and 73.33%，respectively，with corresponding Kappa values of 0.547 and 0.478 (P< 0.05). When using PAX8 positivity，WT-1 positivity，and p53 abnormal expression as diagnostic criteria，the diagnostic rate for HGSOC was 67.97%. Lowering the threshold for p53 overexpression in effusion samples from 70% to 60% enabled the identification of 13 additional cases， increasing the diagnostic rate to 78.13% . Furthermore， combining PAX8 positivity， WT-1 positivity， p53 wild type (with a 60% overexpression threshold)，and diffuse strong p16 positivity with high-grade morphological features of tumor cells raised the diagnostic rate for HGSOC in serous cavity effusion to 85.94%. CONCLUSION： The expression patterns of PAX8 and WT-1 in effusion samples were found to be consistent with those in primary ovarian tumors， whereas expression of p53 and p16 showed only moderate consistency. By lowering the positivity threshold for p53 overexpression from 70% to 60% ， the diagnostic criteria of PAX8 positivity， WT-1 positivity， p53 abnormal expression enhanced the detection rate of HGSOC in effusion cytopathology. Additionally，when p53 was wild-type but PAX8-positive/WT-1-positive/p16 diffusely positive，coupled with high-grade morphological features of tumor cells，the diagnosis of HGSOC was better indicated.

OBJECTIVE： To identify prognostic serological indicators and to establish a nomogram model for predicting prognosis of tongue squamous cell carcinoma (TSCC) patients after surgery. METHODS： Patients with TSCC (n=169) who received surgical treatment in Cancer Hospital of Shantou University Medical College and their clinical information were collected. These patients were randomly divided into training group (118 patients) and verification group (51 patients). In the training group， the prognostic factors related to overall survival (OS) of TSCC patients were screened by univariate and multivariate Cox regression analysis， and a nomogram model was constructed. Akaike information and Bayesian information criteria were used to evaluate the goodness of fit of the nomogram. The bootstrap-corrected of consistency and time-dependent consistency indices were used to evaluate discrimination ability of the nomogram. The predictive ability of the nomogram was evaluated by calibration curve analysis. Decision curve analysis was used to evaluate the decision benefit rate of the model. The above methods were analyzed and verified in the verification group. RESULTS： The prognostic factors related to OS of TSCC patients were pTNM stage， hydroxybutyrate dehydrogenase，blood urea nitrogen，total bilirubin，alanine aminotransferase and factor B. The analysis results of Akaike information and Bayesian information criteria showed that the nomogram had better goodness of fit than pTNM stage. In the training group， the bootstrap-corrected of consistency index of the nomogram was 0.814，higher than that of the pTNM stage of 0.683. The same result also appeared in the verification group， indicating that the nomogram had better discrimination ability than the pTNM stage. The result of the decision curve analysis showed that， compared with pTNM stage， the nomogram had a better net benefit within a reasonable threshold probability range. The calibration curves of the nomogram showed good consistency between the probabilities and observed values in both the training and verification groups. The Kaplan-Meier survival analysis showed that the nomogram predicted the OS of high- and low-risk TSCC patients in both the training and the verification groups (P<0.01). CONCLUSION： Based on clinical characteristics and serological markers，the nomogram might be a novel tool for evaluating the survival probability of TSCC patients.

OBJECTIVE： To analyze correlations between COL5A2 expression and clinical pathological characteristics of gastric cancer patients. METHODS： Among gastric cancer patients，146 of them with radical surgery were recruited. Their tumor and adjacent tissues were taken for immunohistochemical staining to detect the expression of COL5A2 protein and analyze its relationship with clinical pathological indicators，including prognosis of patients. Gastric cancer cell lines (MKN-45 and AGS) were infected with lentivirus carrying shRNA (short hairpin RNA) vector and COL5A2 gene vector to obtain gastric cancer cell strains with stable knockdown and overexpression of COL5A2， respectively. The effect of knocking down and overexpressing COL5A2 on the proliferation of MKN-45 and AGS gastric cancer cells was observed. RESULTS： The positive expression rate of COL5A2 protein in tumor tissues was 39.7% ， which was higher than that in adjacent tissues. The proportions of T3-T4 stage (58.6%)，N1-N3 stage (63.8%)，and stage III (56.9%) in the COL5A2- positive tumor tissues were higher than that in the COL5A2-negative tissues (36.4% ， 36.4% and 29.5% ， respectively，all P<0.05). Survival analysis showed that the recurrence-free survival rate and overall survival rate of patients in the COL5A2-positive group were lower than those in the COL5A2-negative group (P<0.05). Gastric cancer cell lines overexpressing COL5A2 showed significantly enhanced cell proliferation. Knocking down COL5A2 significantly inhibited the proliferation of gastric cancer cells. CONCLUSION： COL5A2 was highly expressed in tumor tissues of gastric cancer patients，indicating later TNM staging and poorer patient prognosis. Overexpression of COL5A2 promoted gastric cancer cell proliferation in vitro. COL5A2 may be useful as a new biomarker for predicting the prognosis of gastric cancer patients.

OBJECTIVE： To investigate the effect of polyacrylamide DNA composites on chromosome aberration of Chinese hamster lung cells (CHL) in vitro， and to evaluate whether it is a mutagen. METHODS： The concentrations of polyacrylamide DNA composites in the chromosome aberration test were set according to the results of the cytotoxicity test (1.25， 2.5 and 5 μL/mL， respectively)， and negative and positive control groups were set up at the same time. The chromosome aberration test was carried out in the presence and absence of the metabolic activation system. After being treated with polyacrylamide DNA composites for different time periods (4 h in the presence of the metabolic activation system，4 and 24 h in the absence of the metabolic activation system)，the cells were collected，and the structure and number of chromosomes were analyzed after hypotonic，fixation，slide preparation and staining. RESULTS： The results of the cytotoxicity test showed that the polyacrylamide DNA composite had no toxic effect on CHL cells. After CHL cells were treated with 1.25， 2.5 and 5 μL/mL polyacrylamide DNA composites， the chromosome aberration rates of cells in the metabolic activation system treatment group were 4% ， 4% and 7% ， respectively. The chromosome aberration rates of cells in the short- term treatment group without metabolic activation were 5%，3% and 7%，respectively，and the chromosome aberration rates of cells in the continuous treatment group without metabolic activation were 4%，2% and 3%，respectively. CONCLUSION： Under the experimental conditions，no aberration-inducing effect of polyacrylamide DNA composite materials on CHL cells was observed.

OBJECTIVE： To determine chromosomal aneuploidy and related factors in first-trimester spontaneous aborted tissues. METHODS： Conducted detection of aneuploid chromosomes and microduplication or micro-deletion of gene fragments which are larger than 100 kb， and retrospective analysis to find the incidence of chromosomal abnormalities in products of conceptions (POC). Compared with existing literature to identify related factors. RESULTS： Among the 56 cases with abnormal karyotypes，46 cases had abnormal number of chromosomes. The number of cases with abnormal chromosomes was higher than that with normal chromosomes. Frequencies of micro-duplicates or micro-deletions of gene fragments were associated with ages of pregnant women (P=0.004). The abnormal rate of these micro-duplicates or micro-deletions increased with maternal age and gravidity (P<0.05). CONCLUSION： Presence of chromosomal abnormalities in early spontaneously aborted tissues indicate that these abnormalities were the main cause of miscarriage in the first trimester. For couples with early spontaneous abortion，cytogenetic examination should be performed on all of POC.