Objective:To investigate the esophageal microecology in patients with esophageal carcinoma (EC), and to compare the difference in esophageal flora between patients with esophageal cancer and healthy people.Methods:From July 2018 to July 2019, at Taihe Hospital, 82 EC patients and 20 age-and gender-matched healthy controls during the same period were selected. The pathology of EC were divided into poorly differentiated (8 cases), moderately differentiated (9 cases) and well differentiated cancers (13 cases) according to the degree of differentiation. The esophageal tissue samples of EC patients and healthy individuals were collected. Sample DNA was extracted and the V4 region of bacterial 16S rRNA was amplified by polymerase chain reaction (PCR). Sequencing was performed by lllumina HiSeq 4000 sequencing platform. Alpha-diversity analysis and principal co-ordinates analysis (PCoA) were performed, and linear discriminant analysis (LDA) of linear discriminant analysis effect size (LEfSe) was used to screen different species. The random forest model was verified by receiver operating characteristic (ROC) curve and the esophageal bacterial phenotype was predicted by BugBase database. Non-parametric Kruskal-Wallis H test and Wilcoxon rank sum test were used for statistical analysis. Results:The Chao1 index of the EC patients was higher than that of healthy controls (362.51(284.29, 646.13) vs. 284.83(244.31, 344.74)), and the difference was statistically significant ( Z=-2.857, P=0.004). The results of PCoA showed that the distance between samples of EC patients and healthy control samples was relatively close, and there was no significant difference in the composition of microecology between the two groups ( P>0.05). The abundance of esophageal Cyanobacteria and Verrucomicrobia of EC patients were both higher than those of healthy controls (0.2% vs. 0.1%, 0.4% vs. 0), while the abundances of esophageal Proteobacteria, SR1 and TM7 phylum of EC patients were lower than those of healthy controls (21.9% vs. 34.2%, 0.1% vs. 0.2%, 0.2% vs. 0.5%), and the differences were statistically significant ( Q=0.090, 0.077, 0.010, 0.026 and 0.001, all P<0.05). The abundances of Clostridia, Elostridiales, Pasteurella, Pasteurellaceae, Eikenella, Actinobacillus and Haemophilus in poorly differentiated patients, moderately differentiated and higher differentiated patients were 28.3%, 24.2% and 17.0%, 28.3%, 24.2% and 17.0%, 3.2%, 0.3% and 5.0%, 3.2%, 0.3% and 5.0%, 0, 1.5% and 0.1%, 0.5%, 0 and 0.7%, 1.3%, 0.2% and 3.9%, respectively, and the differences were statistically significant ( Q=0.579, 0.557, 0.390, 0.711, 0.768, 0.768 and 0.768, all P<0.05). LEfSe analysis showed that the abundances of Fusobacterium, Ruminococcus, Odorbacterium and S24_7 of EC patients were higher than those of healthy controls (21.5% vs. 11.7%, 0.5% vs. 0.1%, 0.1% vs. 0 and 0 vs. 0), and the differences were statistically significant (LDA=2.591, 2.379, 2.790 and 2.927, all P<0.05). The ROC curve confirmed that the random forest model was reliable and the AUC value was 0.92. BugBase database phenotypic prediction showed that the phenotype of esophageal bacteria related to biofilm formation, pathogenic potential, mobile elements, oxygen demand (aerobic, anaerobic and facultative bacteria), and oxidative stress tolerance of EC patients were more abundant than those of healthy controls (all P<0.05). Conclusions:The esophageal flora of patients with esophageal cancer has changed. Fusobacterium, Ruminococcus, Odoribacterium and S24_7 may be potential biomarkers of esophageal flora.

Objective To investigate the role of sodium-hyaluronate-modified calcium peroxide nanoparticles(SH-CaO2 NPs)in inducing pyroptosis in human gastric cancer cells and its possible mechanisms.Methods Transmission electron microscopy(TEM),X-ray diffraction(XRD),infrared spectroscopy,and zeta potential test were used to confirm the synthesis of SH-CaO2 NPs.Cell scratch assay and CCK-8 assay were employed to observe the impacts of SH-CaO2 NPs on the migration and proliferation of human gastric cancer cell line HGC-27.The generation of reactive oxygen species(ROS)was observed with confocal laser scanning microscopy(CLSM)and quantified with flow cytometry in the cells after SH-CaO2 NPs treatment or with pretreatment with ROS inhibitor NAC.Furthermore,the effects of pretreatment of NLRP3 inhibitor(MCC950)and Caspase-1 inhibitor(VX765)on the proliferative activity and on expression of their own and their downstream GSDMD in HGC-27 cells were also investigated with CCK-8 assay,immunofluorescence assay and Western blotting.Results TEM images,XRD,infrared spectroscopy,and zeta potential test confirmed the successful preparation of SH-CaO2 NPs.Cell scratch assay and CCK-8 assay showed that application of SH-CaO2 NPs for 24 h significantly inhibited the proliferation of HGC-27 cells(P＜0.001),while,CLSM and flow cytometry indicated the treatment also promoted the production of ROS(P＜0.001).Pretreatment of ROS inhibitor NAC resulted in up-regulation of NLRP3,and increased expression levels of cleaved Caspase-1 and N-terminal fragment of GSDMD(P＜0.001),while pretreatment of both NLRP3 inhibitor and Caspase-1 inhibitor could reverse the process.Conclusion SH-CaO2 NPs inhibit cell viability of human gastric cancer,which may mediate the inflammatory response and pyroptosis by activating the ROS/NLRP3/Caspase-1/GSDMD signaling pathway.

Objective:To explore the flora characteristics and differences of esophageal tissues between elderly esophageal squamous cell carcinoma (ESCC) patients and young and middle-aged ESCC patients, so as to assist in studying the potential biomarkers of elderly ESCC patients.Methods:In this study, a retrospective study was adopted. 72 ESCC patients diagnosed in Taihe Hospital, Shiyan City, Hubei Province from July 2018 to July 2019 were selected, including 49 patients in the elderly group (≥ 60 years old, 40 males and 9 females), 23 patients in the young and middle-aged group (<60 years old, 21 males and 2 females). In the same period, 20 healthy persons without abnormal gastroscopy in endoscopy center were selected as the control group (aged 35-78 years old, median age 57 years old, 16 males and 4 females). The genomic DNA was extracted from the affected esophageal tissues of patients with ESCC and the middle esophageal samples of the control group. The V4 hypervariable region of bacterial 16SrRNA gene sequence was amplified. Illumina HiSeq sequencing technology was adopted. The flora characteristics of elderly, young and middle-aged ESCC patients was compared and analyzed. QIIME and Rstudio software were used to analyze the sequence data, and nonparametric Kruskal-Wallis test or Wilcoxon rank sum test were used for statistical methods.Results:Shannon index [5.17 (4.53, 5.95) vs. 4.79 (3.74, 5.97)], Simpson index [0.94 (0.91, 0.96) vs. 0.92 (0.83, 0.96)] and Chao1 index [343.55 (259.76, 570.59) vs. 329.16 (268.88, 648.00)] were similar in flora of two groups, and there was no significant difference ( Z=-0.791, -1.057, -0.380, all P>0.05). There was no significant difference in β-diversity between the elderly group and the young and middle-aged group (PC1=19.14%, PC2=6.95%, PPC1=0.67, PPC2=0.42). At the phyla level, the top 5 phyla in abundance were as follows: Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria in the young and middle-aged group, while the top 5 phyla in abundance were as follows: Firmicutes, Bacteroidetes, Proteobacteria, Fusobacteria and Actinobacteria in the elderly group; the significant difference between the two groups was Fusobacteria ( Q=0.596, P<0.05). At the genus level, the top 5 genera in the young and middle-aged group in abundance were as follows: Prevotella, Bacteroides, Streptococcus, Selenomonas and Veillonella. In the elderly group, Prevotella, Bacteroides, Streptococcus, Selenomonas and Haemophilus were the top 5 in abundance, and there were significant difference in Fusobacterium between the two groups ( Q=0.938, P<0.05). PICRUSt function prediction showed that the abundance of Aminoacyl.tRNA.biosynthesis, Nucleotide.excision.repair, RNA.polymerase, Ribosome, Clavulanic.acid.biosynthesis, Photosynthesis and Photosynthesis. proteins in the elderly group were lower than those in the young and middle-aged group (all Q=0.734, P<0.05). Conclusion:There is no significant difference in α-diversity and β-diversity between elderly ESCC patients and young and middle-aged patients, but the abundance of Fusobacterium flora increased.

Objective:To explore the flora characteristics and differences of esophageal tissues between elderly esophageal squamous cell carcinoma (ESCC) patients and young and middle-aged ESCC patients, so as to assist in studying the potential biomarkers of elderly ESCC patients.Methods:In this study, a retrospective study was adopted. 72 ESCC patients diagnosed in Taihe Hospital, Shiyan City, Hubei Province from July 2018 to July 2019 were selected, including 49 patients in the elderly group (≥ 60 years old, 40 males and 9 females), 23 patients in the young and middle-aged group (<60 years old, 21 males and 2 females). In the same period, 20 healthy persons without abnormal gastroscopy in endoscopy center were selected as the control group (aged 35-78 years old, median age 57 years old, 16 males and 4 females). The genomic DNA was extracted from the affected esophageal tissues of patients with ESCC and the middle esophageal samples of the control group. The V4 hypervariable region of bacterial 16SrRNA gene sequence was amplified. Illumina HiSeq sequencing technology was adopted. The flora characteristics of elderly, young and middle-aged ESCC patients was compared and analyzed. QIIME and Rstudio software were used to analyze the sequence data, and nonparametric Kruskal-Wallis test or Wilcoxon rank sum test were used for statistical methods.Results:Shannon index [5.17 (4.53, 5.95) vs. 4.79 (3.74, 5.97)], Simpson index [0.94 (0.91, 0.96) vs. 0.92 (0.83, 0.96)] and Chao1 index [343.55 (259.76, 570.59) vs. 329.16 (268.88, 648.00)] were similar in flora of two groups, and there was no significant difference ( Z=-0.791, -1.057, -0.380, all P>0.05). There was no significant difference in β-diversity between the elderly group and the young and middle-aged group (PC1=19.14%, PC2=6.95%, PPC1=0.67, PPC2=0.42). At the phyla level, the top 5 phyla in abundance were as follows: Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria in the young and middle-aged group, while the top 5 phyla in abundance were as follows: Firmicutes, Bacteroidetes, Proteobacteria, Fusobacteria and Actinobacteria in the elderly group; the significant difference between the two groups was Fusobacteria ( Q=0.596, P<0.05). At the genus level, the top 5 genera in the young and middle-aged group in abundance were as follows: Prevotella, Bacteroides, Streptococcus, Selenomonas and Veillonella. In the elderly group, Prevotella, Bacteroides, Streptococcus, Selenomonas and Haemophilus were the top 5 in abundance, and there were significant difference in Fusobacterium between the two groups ( Q=0.938, P<0.05). PICRUSt function prediction showed that the abundance of Aminoacyl.tRNA.biosynthesis, Nucleotide.excision.repair, RNA.polymerase, Ribosome, Clavulanic.acid.biosynthesis, Photosynthesis and Photosynthesis. proteins in the elderly group were lower than those in the young and middle-aged group (all Q=0.734, P<0.05). Conclusion:There is no significant difference in α-diversity and β-diversity between elderly ESCC patients and young and middle-aged patients, but the abundance of Fusobacterium flora increased.