【Abstract】Objective  To screen ncRNAs involved in the development of small cell lung cancer（SCLC） and predicted potential targets for its diagnosis and treatment. Methods Differential gene analysis was performed on the expression profile microarray data obtained from Gene Expression Omnibus (GEO), ceRNA network was constructed, and functional enrichment analysis and PPI network construction were performed. A total of 7 patients with surgically treated small cell lung cancer were collected, and some differential genes (lncRNA, mRNA and miRNA) were validated in clinical samples using qPCR. Results A total of 99 lncRNAs, 46 miRNAs and 73 mRNA molecules were involved in the construction of the regulatory network. GO and KEGG analyses indicated that the dysregulated mRNAs were mainly involved in cellular cancerization, proliferation and metastasis in small cell lung cancer. According to the PPI network analysis, 46 genes were considered hub genes. qPCR Results  showed that compared to adjacent normal tissues, lnc-ENST00000430027.3 showed relatively high expression (P＜0.001), hsa-miR-143-5p showed relatively low expression (P＜0.01) and FANCA showed relatively high expression (P＜0.01) in small cell lung cancer tissues. Conclusion Combined with the validation of clinical specimens for some genes, it provides a new basis for the molecular biology study of small cell lung cancer diagnosis and treatment.

【Abstract】Objective To investigate the relationship between serum exosome hsa_circ_0004390 and chemotherapy resistance and adverse prognosis in patients with epithelial ovarian cancer (EOC). Methods From January to December 2017, the cancer tissue and serum samples of 87 patients with primary EOC were collected from the First Department of Oncology of Qidong People’s Hospital (QidongHospital affiliated to Nantong University), and 85 healthy female volunteers were recruited to obtain control serum samples. The patients who were ineffective in adjuvant chemotherapy or had disease progression within 6 months were included in the drug resistance group (n=30), and the other patients were included in the sensitive group (n=57). Tissue samples from 10 patients undergoing radical surgery were used for next generation sequencing (NGS) to screen the circRNA gene spectrum. The expression level of candidate circRNAs gene was verified by real-time fluorescent quantitative PCR (qPCR).Results Among the circRNA genes identified by NGS, a total of 285 circrnas expressed differently in the EOC tissues of the drug-resistant group and the sensitive group (P＜0.05). After verification, the relative expression levels of hsa_circ_0004390 (P＝0.004) and hsa_circ_0006276 (P＝0.045) in the EOC tissue of the 5 patients in the drug-resistant group were higher than sensitive group, so we chose these two genes for further analysis. q-PCR showed that compared with the control group, the relative expression of serum exosomalhsa_circ_0004390 [2.57 (1.47, 2.99) vs. 0.94 (0.61, 1.35), P＜0.001] and hsa_circ_0006276 [1.26 (1.03, 1.46) vs. 0.87 (0.67, 1.39), P＝0.001] in EOC group were significantly increased. Further analysis of the working characteristic curve of the subjects found that the serum exosomalhsa_circ_0004390 was more effective in the diagnosis of EOC. In addition, the expression of serum exosomalhsa_circ_0004390 in patients with drug resistance was significantly higher than that in patients with sensitivity group [3.07 (2.66, 3.66) vs. 1.69 (1.23, 2.59), P＜0.001]. However, serum exosomalhsa_circ_0006276 was not associated with platinum chemotherapy reaction (P＞0.05). After further adjusting the Objective  interference factors, it was found that the serum exosomalhsa_circ_0004390 was an independent risk factor affecting the platinum chemotherapy response in EOC patients (P＜0.05). In addition, patients with low level of serum exosomalhsa_circ_0004390 expression (＜2.57) and patients with high level (≥2.57) of OS were better (P＝0.005).Conclusion Serum exosome hsa_circ_0004390 contribute to predict the chemotherapeutic response and survival prognosis of Platinum-based EOC patients, which may provide a new target for EOC treatment.

【Abstract】Objective To investigate the relationship between Ki-67 expression and the clinicopathologic features and prognosis of patients with carcinoembryonic antigen (CEA) and gastric cancer after surgery. Methods Retrospective analysis of clinical and follow-up data of patients who underwent radical gastrectomy for cancer at the Jiangsu Oncology Hospital between January 2015 and December 2016. The serum CEA expression was detected by electrochemiluminescence within 1 week before operation, the expression of Ki-67 was detected by immunohistochemistry. Correlation between Ki-67, CEA expression, and clinicopathologic characteristics was analyzed using the Pearson chi-square test. Kaplan-Meier analysis method, Log-rank test, and Cox regression model were used to analyze the prognostic impact of Ki-67 or combined CEA. Results Of 798 patients with postoperative gastric cancer, 368(46.1%) had Ki-67 high expression. Ki-67 expression was associated with age (P＜0.01), TNM stage (Ⅲ vs. Ⅰ, P＝0.036), T stage (T₄ vs. T1-T3, P＜0.001) and CEA (P＝0.01), but not N stage (P>0.05). CEA correlates with clinical stage (stage Ⅰ-Ⅲ , P＝0.003). The 5-year overall survival (OS) rate for all patients was 68.4%. Cox regression analysis showed that age (P＜0.001), TNM stage (P＜0.001), Ki-67 (P＝0.04), and CEA (P＝0.006) were independent predictors of survival after gastric cancer surgery. The 5-year OS rate of patients with stage Ⅲ with Ki-67-L and CEA-normal-expression (CEA-N) was higher than that of patients with high expression (59.8% vs. 48.9%,P＝0.003;60.4% vs.43.8%,P＝0.003), but no correlation was found in patients with stage Ⅰ and Ⅱ. Statistically significant differences in 5-year OS were observed between low-risk (Ki-67-L + CEA-N) group, intermediate-risk (Ki-67-H or CEA-H) group, and high-risk (Ki-67-H + CEA-H) groups (76.3% vs. 66.7% vs. 55.1%, P＜0.001) group. In patients with stage Ⅱ and Ⅲ, the 5-year OS rate was higher in low-risk group than in the intermediate-risk or high-risk group (78.4% vs. 79.1% vs. 61.4%, P＝0.008;68.2% vs. 47.3% vs. 46.3%, P＜0.001). Conclusion Ki-67 may affect survival in patients with gastric cancer after surgery, combined detection of Ki-67 and CEA contributes to improved predicting prognosis in patients with stage Ⅱ and Ⅲ gastric cancer.

【Abstract】Objective  To investigate the expression and clinical significance of exosomal microRNAs (miR-133a) in clear cell renal cell carcinoma (ccRCC). Methods Serum samples were collected from 293 patients with ccRCC who underwent radical nephrectomy in our hospital from September 2015 to January 2019, and another 200 healthy people were collected as control group. Total RNA was extracted from serum and tissue exosomes, and differentially expressed mirnas were identified by TaqMan low-density microarray (TLDA), and verified by real-time fluorescence quantitative PCR (qPCR). Overall survival (OS) and disease-free survival (DFS) were followed up. Results Compared with healthy control group serum and adjacent normal tissue, three miRNAs (miR-133a, miR-210, miR-7-1) were differentially expressed in serum exosomes and tumor tissues of ccRCC patients by TLDA screening. The expression of exosomes miR-133a was significantly down-regulated in ccRCC patients by qPCR (P＜0.001). The expression of exosome miR-133a in ccRCC patients was positively correlated with that in tumor tissues (r=0.467, P＜0.001). Receiver operating characteristic (ROC) curves confirmed that the area under the ccRCC curve (AUC) for the diagnosis of serum exosome miR-133a was 0.78 (95%CI: 0.747-0.729). The expression of preoperative serum exosome miR-133a was correlated with TNM stage and Fuhrman grade (P＜0.05), but not renal function index (P>0.05). The DFS and OS of patients with high expression of serum exosome miR-133a before surgery were both longer than those with low expression (P＜0.001). Cox proportional regression analysis showed that serum exosome miR-133a was an independent prognostic factor for OS in patients with ccRCC [HR=0.486 (95CI: 0.226-1.043); P＝0.034]. Conclusion Serum exosome miR-133a is closely related to the occurrence, development and prognosis of ccRCC, and may be a biomarker to predict the prognosis of patients with ccRCC.

【Abstract】Objective To analyze the expression levels of platelet membrane glycoprotein Ⅱb/Ⅲa fibrinogen receptor (PAC-1), platelet P-selectin (CD62P) and SLC7A11 on platelet surface and their clinical significance in elderly patients with laryngeal squamous cell carcinoma. Methods Eighty-seven elderly patients with laryngeal squamous cell carcinoma admitted to our hospital from January 2016 to January 2021 (laryngeal cancer group) were collected, and 87 patients with vocal cord polyp admitted to our hospital during the same period (control group) were selected. Blood and pathological specimens were collected from the patients, and the positive rate of specific fluorescent antibody binding platelets was determined by flow cytometry using monoclonal immunofluorescence labeling, PAC-1 and CD62P were detected, and expression of SLC7A11 was detected by immunohistochemical SP method. The relationship between the expressions of PAC-1, CD62P and SLC7A11 and the prognosis of patients was analyzed. Results The positive expression rate of PAC-1 and CD62P in laryngeal cancer group was higher than that in control group, and the high expression rate of SLC7A11 (62.07%) was lower than that in control group (28.74%), the difference was statistically significant (P＜0.05). Among the 87 patients with good prognosis, 69 patients had good prognosis, and 18 patients had poor prognosis. The positive expression rate of PAC-1 and CD62P in the poor prognosis group was higher than that in the good prognosis group, and the high expression rate of SLC7A11 (94.44%) was higher than that in the good prognosis group (53.62%) (P＜0.05). The proportion of clinical stage and cervical lymph node metastasis in patients with poor prognosis was significantly higher than that in the control group (P＜0.05), but there were no differences in age, gender and degree of differentiation between the two groups (P>0.05). Multivariate Logistic regression analysis with clinical stage, cervical lymph node metastasis, PAC-1, CD62P and SLC7A11 as independent variables showed that clinical stage, cervical lymph node metastasis, PAC-1, CD62P and SLC7A11 were independent risk for recurrence (P＜0.001). Conclusion PAC-1, CD62P and SLC7A11 are highly expressed in elderly patients with laryngeal squamous cell carcinoma, and are closely related to the prognosis of patients, which may be used as a reference indicator for prognosis evaluation.

【Abstract】Objective  To investigate whether circDUSP16 can be used as a potential biomarker to predict the efficacy and prognosis of adjuvant chemotherapy for resectable gastric cancer (GC). Methods Serum samples were collected from 226 GC patients who underwent radical gastrectomy in our hospital from January 2017 to May 2021. In addition, the serum samples of 209 healthy people in our hospital were selected as control. The expression level of circDUSP16 in serum was detected by real-time quantitative PCR (qPCR). Patients with recurrence or metastasis within 6 months after the end of chemotherapy were included in the chemotherapy insensitive group, while others were included in the chemotherapy sensitive group. Results Serum circDUSP16 level in GC group was significantly higher than that in non-cancer control group (P＜0.001). The area under ROC curve (AUC) of serum circDUSP16 for GC diagnosis was 0.874 (95%CI: 0.843-0.906). 95 patients (42.0%) with GC relapsed within 6 months after the end of chemotherapy. The expression level of circDUSP16 in serum of chemotherapy insensitive group was significantly higher than that of chemotherapy sensitive group (P＜0.001). The AUC of serum circDUSP16 predicted chemotherapy sensitivity was 0.898 (95%CI: 0.854-0.941), and the specificity and sensitivity were 89.8% and 80.8%, respectively, at the optimal cut-off value of 1.870. Multivariate Logistic regression analysis showed that serum circDUSP16 was an independent factor for chemotherapy sensitivity in GC patients (P＜0.001). According to the median value of circDUSP16 in serum of GC patients, the patients were divided into low expression group (< 1.64) and high expression group (≥1.64). The overall survival rate of baseline GC patients in the low and high circDUSP16 expression groups was 69.9% and 14.2%, respectively, and the median OS stage was longer in the low expression group (P＜0.001). Multivariate Cox proportional regression analysis showed that serum circDUSP16 was an independent prognostic factor for GC patients (P＜0.001). Conclusion High serum circDUSP16 expression was associated with drug resistance and poor survival prognosis of GC adjuvant chemotherapy, which is promising as a potential predictor of GC adjuvant chemotherapy efficacy and poor prognosis.

【Abstract】Objective  To investigate the clinical significance of miR-204 promoter methylation in colorectal cancer (CRC). Methods From January 2018 to March 2020，the blood samples, tumor tissue samples and paracarcinoma normal colon mucosal tissue samples were collected from 69 CRC patients admitted to the department of gastroenterology of our hospital. Another 68 healthy volunteers were recruited as the normal control group. The plasma samples were collected. Methylation levels of CpG sites identified in tumor and normal tissue were analyzed using the Tumor Gene Atlas Project (TCGA) database and gene-on-a-chip data (Methyl450K). The gene expression and promoter methylation status of miR-204 were detected by quantitative methylation-specific PCR (qMSP) and quantitative real-time PCR (qPCR). Results The TCGA data indicated that the expression of miR-204 was coordinately down-regulated with the expression of host gene TRPM3, and two CpG islands around the miR-204 promoter were identified. Compared with the paracancer tissues, the miR-204 promoter methylation reference percentage (PMR) was increased in CRC tissues[1.78 (0.35, 16.61) vs. 1.51 (0.45, 2.98), P＜0.001]. And the methylation level of miR-204 promoter was negatively correlated with the ex