OBJECTIVE：The purpose of this study was to improve diagnostic efficacy of urothelial carcinoma (UC) by using the droplet digital PCR (ddPCR) to detect FGFR3 S249C (fibroblast growth factor receptor 3，FGFR3)mutation in cfDNAs which were extracted from urine of UC patients and in combination with urinary cytology. METHODS：Urine samples were collected from 27 UC patients and DNA samples were extracted. The FGFR3 S249C mutation in urinary cfDNA was detected by ddPCR. Receiver operating characteristic curves (ROC) were applied to evaluate the diagnostic efficacy of urinary cfDNA for patients with unclear urinary cytology but a clear pathological diagnosis of UC. RESULTS：The detection rate of the FGFR3 S249C mutation for UC was 25.9% (7/27). The detection rate of the mutation classified by TPS (The Paris system for reporting urinary cytology) was 18.8% (3/16) in HGUC (high-grade urothelial carcinoma)，50% (4/8) in SHGUC (suspicious for high-grade urothelial carcinoma)，and 0 (0/3) in NHGUC (negative for high-grade urothelial carcinoma). The mutation rate was 25.0% (1/4) in non-invasive UC and 31.6%(6/19)in invasive UC. The detection rate of positive mutation by ddPCR in SHGUC was 50.0% (4/8)，and the area under the ROC for the diagnosis of UC by the mutation was 0.781 with 95% CI (0.407，1.000)，which showed good accuracy for its diagnosis. CONCLUSION：Measurement of the FGFR3 S249C mutation in urinary cfDNA using ddPCR as a non-invasive assay could improve the diagnostic efficacy for UC with unclear urinary cytology diagnosis.

OBJECTIVE：Expression of immune cell infiltration (ICI) and tumor mutation burden (TMB) in colorectal cancer (CRC) samples were analyzed using bioinformatic analysis. METHODS：The CIBERSORT and ESTIMATE algorithms were used to calculate relative fractions of immune cells in CRC samples，and the Spearman test was used to analyze correlations among infiltrating immune cells. The ConsensusClusterPlus was used to cluster the CRC samples into ICI cluster A and ICI cluster B based on similarity of the relative fractions of immune cell infiltration. The overall survival (OS) of patients were analyzed by Kaplan-Meier. The ICI score model of CRC samples was constructed based on the signature gene A and signature gene B，and ICI scores of each CRC samples were calculated. CRC patients were divided into two groups based on the ICI scores：ICI score High and Low. The OS of patients were analyzed by Kaplan-Meier，and GSEA was used to analyze the enrichment pathways in each group. TMB was calculated and presented by maftool. Based on the TMB，the CRC patients were divided into H-TMB and L-TMB. The OS of patients between two groups were analyzed by Kaplan-Meier. RESULTS：There was no statistical difference in OS between patients in ICI clusters A and B. However，patients in the ICI score high group had longer OS than those in the ICI score low group. Gene sets which were closely related to intestinal physiological functions were enriched in the ICI score high samples，while gene sets which were closely related to tumor formation，development and metastasis were enriched in the ICI score low group. There was no significant difference in TMB between ICI score low and ICI score high samples. The frequencies of KRAS，TP53 and APC were the highest compared with that of other genes in both the ICI score low and high groups. The patients in the L-TMB group had longer OS than those in the H-TMB group. CONCLUSION：CRC patients in the ICI score high group had longer OS than those in the ICI score low group，and patienets in the L-TMB group had longer OS than those in the H-TMB group.

OBJECTIVE：Relationships between DNA ploidy abnormality and epidermal growth factor receptor (EGFR) gene mutation in pleural effusion from metastatic lung adenocarcinomas and their effects on prognosis of patients. METHODS：Clinical data of 439 patients with lung adenocarcinomas and with malignant pleural effusions were collected from the Fourth Hospital of Hebei Medical University from 2012 to 2019. DNA content in cells from pleural effusions was detected by computer aided DNA ploidy quantitative analysis. Percentages of cells with greater than 5c，the number of cells with greater than 9c，the maximum DNA index (DI) value，the average DI value of cells with greater than 5c and the peak of aneuploid cells were calculated. Differences of DNA ploidy and EGFR gene mutation among groups were compared，and correlations between DNA ploidy and EGFR mutation were tested to analyze the values of DNA ploidy and EGFR gene mutation in prognosis of lung adenocarcinoma. RESULTS：EGFR mutations were detected in 244 (55.58%) of 439 patients. The mutation rate was significantly reduced in male patients，and in patients with a history of smoking，compared to the controls (P<0.05). In addition，the percentage of cells greater than 5c <14%，the number of cells greater than 9c <8，the maximum DI value <6，and the mean value of cells greater than 5c <5 (P<0.05). The mutation rate of EGFR gene was different with different aneuploid cell peak，and compared with the nonpeak group，the mutation rate of EGFR gene was higher with single and double peak (P<0.05)，while there was no significant difference in EGFR mutation rate between multi-peak and other groups. Compared with the EGFR wild-type patients，EGFR mutant patients had a higher mean rank in the percentage of cells greater than 5c，the number of cells greater than 9c，the maximum DI，the mean DI of cells greater than 5c，and the aneuploid cell peak number，and all five indexes were weakly positively correlated with EGFR mutation. Univariate survival analysis showed that the median overall survival time of patients with greater than 5c cell percentage ≥14%，greater than 9C cell number ≥8 and EGFR mutation were longer than that of the controls. Multivariate survival analysis showed that DNA ploidy was not associated with patient prognosis，and EGFR gene mutation was an independent prognostic factor in patients with advanced lung adenocarcinoma. CONCLUSION：There was a positive correlation between EGFR gene mutation and DNA ploidy abnormality in patients with advanced lung adenocarcinoma，and DNA ploidy abnormality was a potential predictor of EGFR gene mutation. DNA ploidy was not an independent prognostic factor in patients with advanced lung adenocarcinoma. Patients with EGFR gene mutation had a better prognosis than the wild-type patients.

OBJECTIVE：To investigate effects of bisphenol S on expression of spermiogenesis genes in Caenorhabditis elegans (C. elegans). METHODS：Nematodes at L4 stage were exposed to bisphenol S for 24 h (with the concentrations of 0，3，30，300 μmol/L based on the acute toxicity test)，brood size and expression levels of akt-1，swm-1，try-5，spe-4 and spe-6 genes were detected by real-time fluorescence quantitative PCR. RESULTS：After the 24 h exposure to bisphenol S，the LC₅₀ was determined to be 2 773.672 μmol/L. Compared to the control group，exposure with the concentrations of 3，30 and 300 μmol/L significantly reduced the brood size (P<0.05 or P<0.01). In addition，the mRNA level of akt-1 was increased at the concentration of 3 μmol/L (P<0.05)；the expression level of swm-1，try-5，spe-4 and spe-6 mRNA were increased in the 30 and 300 μmol/L treatment groups (P<0.05 or P<0.01). CONCLUSION：Expressions of spermiogenesis genes in C. elegans were affected from exposure to bisphenol S.

OBJECTIVE：To investigate effects of small molecule inhibitor ICG-001 on malignant phenotypes of esophageal squamous carcinoma (ESCC) cells and to explore its molecular mechanisms. METHODS：The ESCC cell lines KYSE410 and KYSE450 were treated with ICG-001 of 50 and 100 μmol/L，or with solvent DMSO as control. Treated cells were evaluated for cell proliferation viability，colony formation ability，cell cycle distribution and apoptosis. mRNA and protein expression of related molecules (SKP2，Survivin，and TCF4) were detected by real-time PCR (qPCR) and Western blot. RESULTS：Compared with the control group，ICG-001 treatment significantly inhibited the proliferation and colony formation ability of KYSE410 and KYSE450，and significantly induced G0/G1 phase arrest (P<0.01). In addition，expression levels of SKP2、Survivin and TCF4 were significantly decreased (P<0.01). CONCLUSION：ICG-001 significantly inhibited the proliferation viability and colony formation ability，and induced G0/G1 phase arrest of the two ESCC cell lines. The mechanisms may be related to the inhibition of b-catenin/TCF4 transcriptional activity and the expression of downstream molecules：Survivin and SKP2.

OBJECTIVE：To investigate diagnostic values of human serum cysteine protease inhibitor S (CST4)，squamous cell carcinoma antigen (SCC)，carbohydrate antigen 199 (CA19-9) and soluble cytokeratin component (CYFRA21-1) in oral and maxillofacial malignant tumors. METHODS：A total of 232 patients with oral and maxillofacial malignancies tumors who were admitted to our hospital from January 2022 to November 2022，and 40 healthy physical examination subjects were selected during the same period as the research objects. Serum levels of CST4，SCC，CA19-9 and CYFRA21-1 from them were analyzed. ROC curve was developed to evaluate diagnostic efficacy of the different indexes. RESULTS：The serum median levels of CST4，SCC and CYFRA21-1 in the malignant tumor group were significantly higher than those in the benign tumor group and the healthy control group (P<0.05). There was no difference in expression levels of CST4 in patients with malignant tumors at different sites and pathological types (P>0.05). ROC curve showed that serum CST4 detection had a high diagnostic value for the tumors (AUC=0.954，sensitivity 97.5%，specificity 78.45%，cut off value 74.68 ng/mL). However，the combined detections of CST4，SCC and CYFRA21-1 were more effective and specific in the clinical diagnosis of the malignant tumors. CONCLUSION：Serum CST4，SCC and CYFRA21-1 were highly expressed in oral and maxillofacial malignant tumors. Compared with the single detection of serum tumor markers，the combined detection of the three had higher reference value in the diagnosis of malignant tumors，and can be recommended as useful tumor markers.

OBJECTIVE：To observe expression levels of serum exocrine miR-193a and miR-21-5p in patients with osteosarcoma，and to explore their diagnostic and prognostic values. METHODS：A total of 112 osteosarcoma patients who visited our hospital from January 2016 to December 2021 were collected as the study subjects，and 100 healthy people as the controls. From all individuals，5mL peripheral blood were collected from each，and serum samples were isolated for exosomes collection. Expression levels of miR-193a and miR-21-5p were detected by real-time quantitative PCR (qPCR). ROC curves were used to evaluate diagnostic values of miR-193a and miR-21-5p for osteosarcoma，and to analyze their correlation with clinical features and prognosis. RESULTS：Expression levels of miR-193a and miR-21-5p from patients were significantly higher than those from controls (P<0.05). The osteosarcoma diagnosis AUC of miR-193a and miR-21-5p were 0.811 and 0.880，respectively. Their sensitivities were 89.2% and 90.2%，and the specificities were 75.0% and 84.0%，respectively. The osteosarcoma diagnosis AUC of the combined method was 0.901，which was higher than that of the single index. The sensitivity and specificity were 92.1% and 86.5%，respectively. Osteosarcoma patients with high expression levels of miR-193a and miR-21-5p were often associated with advanced TNM stages，risk of distant organ metastasis and worse prognosis (P<0.05). CONCLUSION：The expression levels of miR-193a and miR-21-5p in the serum exosomes from osteosarcoma patients were significantly increased. The increase was associated with advanced TNM stages，higher risk of distant organ metastasis and worse prognosis. Thus，serum secretions of miR-193a and miR-21-5p can be considered as biomarkers for diagnosis and prognosis of osteosarcoma.

OBJECTIVE：To explore effects of microRNA-495 (miR-495) on malignant phenotype of esophageal squamous carcinoma cells and its molecular mechanisms. METHODS：miR-495 mimics and miR-495 inhibitors，and the corresponding random sequence control were transfected into esophageal cancer cells Eca109. Real-time fluorescent quantitative polymerase chain reaction PCR (qPCR) was used to detect expression of miR-495，CCK-8 to detect cell proliferation，flow cytometry to detect cell cycle and apoptosis，transwell to detect cell migration and invasion ability，qPCR and western blot to detect expression of mRNAs and proteins of downstream target gene RNF113A. RESULTS：Compared with the control group，the miR-495 mRNA levels in the miR-495 mimics transfected group were significantly increased (P<0.05)，and in the miR-495 inhibitor transfected group was significantly decreased (P<0.05). There was no significant changes in cell proliferation among the groups (P>0.05). The migration and invasion numbers in the miR-495 mimics transfected group were significantly decreased (P<0.01)，and the apoptosis rate was significantly increased (P<0.01). The levels of cells blocked in the G0/G1 phase (P<0.01) were significantly more than that in the controls. The migration and invasion numbers in the miR-495 inhibitor transfected group were significantly increased (P<0.01)，and the apoptosis rate was significantly decreased (P<0.01). The proportion of S phase cells in the miR-495 inhibitor transfected group was increased (P<0.05)，and the difference was statistically significant. Compared with the control group，expression of the RNF113A protein in the miR-495 mimics transfected group was down-regulated (P<0.05)，and expression of the RNF113A in the miR-495 inhibitor transfected group was up-regulated (P<0.05). However，the mRNA levels of RNF113A did not change significantly (P>0.05). CONCLUSION：miR-495 promoted cell apoptosis，inhibited cell migration and invasion ability，and regulated cell cycle distribution. Thus，miR-495 may mediate malignant phenotype of esophageal squamous carcinoma cells by regulating expression of the RNF113A protein.

OBJECTIVE：To investigate expression of the splicing factor 2/alternative splicing factor (SF2/ASF) in pancreatic cancer tissues and its correlation with oncogene P53 (mutant type)，proliferation marker Ki67 and clinicopathological features. METHODS：Differences in expression of the SRSF1 gene (the gene encoding SF2/ASF) in 350 pancreatic cancer tissues and normal tissues adjacent to the cancer were evaluated based on GEPIA database analysis；and correlations with SRSF1 and TP53 and MKi67 were analyzed. In addition， expression of SF2/ASF，mutant P53 and Ki67 protein in 60 pancreatic cancer tissues and corresponding normal tissues were evaluated，as well as correlations between SF2/ASF and mutant P53 and Ki67 using Pearson correlation analysis，and correlations between SF2/AFS protein expression levels and clinicopathological indices of pancreatic cancer patients，and prognostic analysis using Cox proportional risk regression model. RESULTS：Results from our database analysis show that expression of the SRSF1 gene was significantly higher in pancreatic cancer tissues compared with normal tissues (P<0.01)，and expression of the SRSF1 was correlated with both TP53 and MKi67 (r=0.28 and 0.32，respectively，both P<0.01). Results from the immunohistochemical assay and the Pearson correlation analysis show that SF2/ASF was correlated with mutant P53 protein and Ki67 proliferation index (r=-0.386 and 0.275，respectively，P<0.05) in pancreatic cancer tissues. The chi-square test show that expression of the SF2/ASF in pancreatic cancer was correlated with the degree of pancreatic cancer differentiation (P<0.01)，whether with lymph node metastasis (P<0.01) and the presence of distal organ metastasis (P<0.01)，but not with the site of pancreatic cancer occurrence and TNM stage (P>0.05). Cox risk regression analysis show that high expression of SF2/ASF and patient age (both P<0.01) were independent risk factors for overall patient survival. CONCLUSION：SF2/ASF，mutant P53 and Ki67 proteins were highly expressed in pancreatic cancer，and SF2/ASF correlates with mutant P53 and Ki67. Our findings suggest that SF2/ASF can serve as a new target for pancreatic cancer diagnosis and treatment.

OBJECTIVE：To understand the impact of job type，gender，and employment duration on exposure doses of radiation workers in medical institutions，and to compare efficiency of chromosomal aberration analysis and thermoluminescence monitoring in assessing occupational health of the workers. METHODS：By using logistic regression analysis，the chromosomal aberration rates from different groups of radiation workers were analyzed based on three variables：job type，gender，and employment duration. The ROC curve was constructed based on individual dose monitoring results. RESULTS：Chromosome aberration rates of nuclear medicine workers were significantly higher than those who carried out X-ray imaging diagnosis (P<0.05)，while the rates for radiation therapy and interventional radiology workers were not statistically significant (P>0.05). There was no significant (P>0.05) impact from gender and employment duration on the chromosome aberration rates. The consistency between the chromosome aberration analysis and the abnormal rates of thermoluminescence monitoring (assuming that it exceeded 1 mSv as abnormal) was good. CONCLUSION：The radiation exposure doses of staff in the nuclear medicine department were significantly higher than that of staff in radiation diagnosis，radiotherapy and intervention. The evaluation criteria for the two means of occupational health monitoring (personal dose monitoring and chromosome aberration analysis) of radiation workers were inconsistent.

OBJECTIVE：To perform a systematic genotoxicity assessment of fungal fibrinolytic compound 1 (FGFC1) in an established GLP laboratory platform. METHODS：A combination of classical genotoxicity assays (Ames，in vitro culture CHO cell chromosome aberration and mouse bone marrow micronucleus assays) was used to detect genotoxicity of FGFC1. RESULTS：Results from the Ames assay indicate that FGFC1 was not mutagenic to Salmonella typhimurium at five doses of 0.5，5，50，500 and 1 000 g per dish，with and without the addition of the S9 metabolic activation system. FGFC1 did not induce chromosomal aberrations in the presence of S9 after 24 h and 48 h of treatments. The rates of micronucleus induction in bone marrow cells at 3 doses of 62.5，125.0 and 250.0 mg/kg was not significantly different from that in the solvent control group (P>0.05). CONCLUSION：The above results indicate that FGFC1 was not genotoxic and not potentially carcinogenic under the test conditions.