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 study the effect of epithelial cell transformation sequence 2 (ECT2) on the proliferation of cervical cancer cells and its mechanism. Methods We transfected cervical cancer cells HeLa (HeLa-ECT2) with the lentivirus overexpressing ECT2 and the cells SiHa (SiHa-siRNA) and C33a (C33a-siRNA) with the interfering plasmid. MTT assay was performed to detect cell proliferation ability. Flow cytometry was conducted to detect the cell cycle of each group. The IPA database was searched for the interacting proteins of ETC2, and immunofluorescence subcellular localization verified the effect between the two. qPCR and Western blot were carried out to detect the expression of Rac1, Cdc42, CDK1, and Cyclin B1 mRNA and protein in each group of cells. Results ECT2 may interact with CDK1. After ECT2 expression was upregulated, the G₂/M phase of HeLa-ECT2 cells accelerated the transformation to G₁ phase, cell proliferation ability was enhanced, and the expression levels of Rac1, Cdc42, CDK1, and cyclin B1 mRNA and protein all increased (P < 0.001); the knockdown of ECT2 expression would reverse the effect (P < 0.05). Conclusion ECT2 accelerates G₂ phase of cervical cancer cells to G₁ phase and promotes cell proliferation by co-localizing with CDK1 through the downstream Cdc42/Rac1 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.

Objective: To explore the expression levels of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) protein and the change of MMP-9/TIMP-1 ratio in wound exudates of patients with stages Ⅲ and Ⅳ pressure ulcers during wound healing. Methods: From July 2017 to July 2018, 30 patients with stage Ⅲ pressure ulcers [30 wounds, 16 males and 14 females, aged (65±10) years] and 34 patients with stage Ⅳ pressure ulcers [50 wounds, 17 males and 17 females, aged (65±9) years] admitted to Hebei General Hospital who met the inclusion criteria were enrolled in this prospective cohort study. According to the principle of wound treatment and the characteristics and needs of wound in different periods, individualized intervention measures were formulated for patients and appropriate dressings were selected. At the time of admission and on 7, 14, 21, 28 days of treatment, the healing of pressure ulcer wounds was evaluated by Pressure Ulcer Healing Scale. Afterwards, the wound exudate was collected at each time point to detect the expression levels of MMP-9 and TIMP-1 protein by enzyme-linked immunosorbent assay, and the MMP-9/TIMP-1 ratio was calculated. Data were processed with analysis of variance for repeated measurements of single group and linear trend test. Results: (1) There were significantly statistical differences in wound healing scores of patients with stages Ⅲ and Ⅳ pressure ulcers among the time of admission and on 7, 14, 21, 28 days of treatment within each stage (F=145.382, 153.234, P<0.01), and they all showed a gradually decreasing trend (F=170.466, 284.585, P<0.01). (2) At the time of admission and on 7, 14, 21, 28 days of treatment, the expression levels of MMP-9 protein in wound exudates of patients with stages Ⅲ and Ⅳ pressure ulcers were (171±104), (138±88), (110±70), (85±55), (62±41) ng/L and (193±107), (173±104), (139±83), (114±70), (89±56) ng/L, respectively. There were significantly statistical differences within each stage (F=58.007, 111.680, P<0.01), and they all showed a gradually decreasing trend (F=62.901, 134.628, P<0.01). At the time of admission and on 7, 14, 21, 28 days of treatment, the expression levels of TIMP-1 protein in wound exudates of patients with stages Ⅲ and Ⅳ pressure ulcers were (6.2±3.9), (5.6±3.4), (5.1±3.1), (4.4±2.5), (3.8±2.3) ng/L and (4.8±2.5), (4.7±2.6), (4.4±2.6), (4.6±2.7), (4.1±2.4) ng/L, respectively. There were significantly statistical differences within each stage (F=25.479, 7.778, P<0.01), and there was a gradually decreasing trend in stage Ⅲ (F=62.901, P<0.01) and a decreasing trend in stage Ⅳ (F=134.628, P<0.01). At the time of admission, the expression levels of MMP-9 and TIMP-1 in wound exudates of patients with stage Ⅲ pressure ulcers were similar to those of patients with stage Ⅳ pressure ulcers (t=-1.03, 1.47, P>0.05). (3) At the time of admission and on 7, 14, 21, 28 days of treatment, the MMP-9/TIMP-1 ratios in the wound exudates of patients with pressure ulcers of stages Ⅲ and Ⅳ were 30±13, 25±9, 22±9, 20±8, 17±6 and 43±19, 37±13, 32±10, 26±9, 22±9, respectively. There were significantly statistical differences within each stage (F=37.173, 97.191, P<0.01), and they all showed a gradually decreasing trend (F=54.183, 130.088, P<0.01). At the time of admission, the MMP-9/TIMP-1 ratio in wound exudates of patients with stage Ⅳ pressure ulcers was significantly higher than that of patients with stage Ⅲ pressure ulcers (t=-3.42, P<0.01). Conclusions During the wound healing process of patients with stages Ⅲ and Ⅳ pressure ulcers, the expression levels of MMP-9 and TIMP-1 protein and the MMP-9/TIMP-1 ratio in wound exudates show a decreasing trend. The stage of wound healing can be predicted according to the expression level of MMP-9 protein and the MMP-9/TIMP-1 ratio.

Chronic wounds are with characteristics of long last time and cannot heal in time, which is a problem in clinic. Wound pH value plays an important role in the process of healing of chronic wounds. In this paper, we review the relative researches on wound pH value and wound microenvironment, summarize the potential relationship between wound pH value and healing of chronic wounds, as well as the method to change pH value of chronic wounds, thereby to provide theoretical basis for the treatment of chronic wounds in clinic.