Objective  To investigate the expression of microRNA-1253（miR-1253） in lung adenocarcinoma cell lines and its effect on the proliferation， migration and invasion of lung adenocarcinoma cells. Methods  Real-time fluorescence quantitative PCR （QPCR） was used to detect the expression of miR-1253 in lung adenocarcinoma cell lines A549， NCI-H1299， NCI-H157， A973 and GLC-82. miR-1253 mimics and miR-1253 inhibitor were transfected into A973 and NCI-H157 cells respectively. The cells transfected with negative control plasmid NC were used as negative control（NC） group. The effects of different expression of miR-1253 on proliferation， clone formation， invasion and migration of A973 and NCI-H157 cells were examined by cell proliferation assay, clone formation assay and Transwell migration and invasion assay. Results  The results of QPCR showed that the relative expression of miR-1253 in A549， NCI-H1299， NCI-H157， A973 and GLC-82 cells were 0.92±0.06, 0.06±0.03, 1.10±0.26, 0.03±0.01 and 0.45±0.08， respectively. After transfection of miR-1253 mimics into A973 cells， the relative expression of miR-1253 increased significantly（P＜0.05）， and decreased significantly after transfection of miR-1253 inhibitor into NCI-H157 cells （P＜0.05）. Compared with the NC group， the transfection of miR-1253 mimics could significantly inhibit the proliferation activity， the ability of plate cloning（166.0±29.3 vs. 371.0±31.4， P=0.001） and the ability of migration（91.1±32.1 vs. 166.7±33.9， P=0.008） and invasion（74.4±20.5 vs. 145.6±28.8，P=0.001） of lung adenocarcinoma cell line A973； miR-1253 inhibitor can up-regulate the proliferation， number of plate cloning cells（545.0±61.9 vs. 337.0±39.7， P=0.008）， migration（246.7±36.7 vs. 151.1±32.9，P＜0.001） and invasion（231.1±38.8 vs. 137.8±27.3， P=0.001） of NCI-H157. Conclusion  miR-1253 could reduce malignant phenotype of lung adenocarcinoma cells， and therefore it can be used as an effective target for gene therapy of lung adenocarcinoma.

Objective  To investigate the effect of Fasudil on the pfoliferation， invasion and migration of gastric cancer MGC-803 cells and their probable mechanisms. Methods  MGC-803 cells were treated with different concentration of Fasudil and cell number was counted. And the subsequent experiments were conducted in 3 groups respectively contained 10， 25， 50 μmol／L of Fasudil, the MGC-803 cells treated with Fasudil were taken as the treatment group， while those not treated with Fasudil were taken as the control group. The cells were cultured for 4 days（0， 24， 48， 72， 96 h）， and the cell growth folds were detected by CCK-8.The ability of cell migration was detected by wound-healing assay. The ability of cell invasion was detected by Transwell. The protein expression levels of vascular endothelial growth factor （VEGF）， matrix metalloproteinases 9（MMP-9）， matrix metalloproteinases14 （MMP-14） and epithelial mesenchymal transformation （EMT） markers Vimentin and E-cadherin were detected by Western blotting. Results  The toxicity test showed that Fasudil inhibited the cell growth of MGC-803 cells in a dose-dependent manner and the cell growth folds were reduced in the treatment group. There was an obvious decrease in the growth folds compared to the control group （P＜0.01）. The results of wound healing assay showed that the wound closure rate in the treatment group was lower than that of the control group， and in a dose-dependent manner（P＜0.01）. Transwell results showed that the number of invasive cells in the treatment group was decreased significantly compared with the control group （P＜0.01）. Besides， Fasudil significantly decreased the expression levels of VEGF， MMP-9， MMP-14 and Vimentin， and up-regulated the expression levels of E-cadherin. Conclusion  Fasudil may inhibit cell proliferation， migration and invasion of MGC-803 cells， reduce the activity of MMP-9， MMP-14 and VEGF， which will provide new sight and theoretical basis for the treatment of gastric cancer.

Objective  To investigate the expression of ADAM15 in colorectal cancer cells and its effect on migration and invasion of colorectal cancer cells.

MethodsThe expression of ADAM15 protein in human normal colorectal mucosal cell line FHC and human colorectal cancer cell line Caco-2， SW-480 and DKO-1 was detected by Western blotting. SW-480 cells were transfected into siNC （siNC group） and siADAM15 （siADAM15 group） respectively. Cell scratch test and Transwell test were used to detect the migration and invasion ability of the two groups. The expressions of pEGFR， EGFR and ADAM15 proteins in siNC， siADAM15 and FHC were detected Western blotting. Results  The expression levels of ADAM15 in Caco-2， SW-480 and DKO-1 cell lines were 2.01±0.149， 2.74 ±0.053 and 2.27 ±0.185， respectively， which were higher than 0.98±0.043 in FHC cell line， and the difference was statistically significant （P＜0.05）. Scratch test showed that the ratio of 24 h scratch width to 0 h scratch width in siADAM15 group was 0.925±0.087， which was significantly higher than 0.326±0.031 in siNC group， and the difference was statistically significant （P＜0.05）. Transwell test showed that the invasive number of SW-480 cells in siADAM15 group was 327.4±2.13， which was significantly lower than 669.7±35.21 in siNC group （P＜0.05）. The expression of p-EGFR in SW-480 cells of siNC group was 0.89±0.097， which was significantly higher than 0.42 ±0.035 in FHC cell line， and the difference was statistically significant （P＜0.05）. The expression of p-EGFR in SW-480 cells of siADAM15 group was 0.27±0.067， which was significantly lower than 0.89±0.097 of siNC group， and the difference was statistically significant （P＜0.05）. Conclusion  ADAM15 may promote the migration and invasion of colorectal cancer cells by activating EGFR.

Objective  To investigate the effect of RNA interference （siRNA） on ribosomal protein S6 kinase 2 （S6K2） expression on proliferation and apoptosis of cervical cancer cells and phosphatidylinositol 3 kinase （PI3K）／protein kinase B （Akt）／nuclear factor-κB （NF-κB） signaling pathway. Methods  The expression of S6K2 in human cervical cancer tissues was analyzed by Oncomine （tumor microarray database）. Human cervical cancer cell lines HeLa and Caski were transfected with siRNA targeting S6K2 （siRNA-S6K2） and control random sequence （siRNA-Ctrl）， respectively. Real-time fluorescence quantitative PCR （QPCR） was used to detect the mRNA levels of S6K2 in each group. CCK-8 method was used to detect the cell proliferation. Annexin V-FITC／PI doublestaining flow cytometry was used to detect the cell apoptotic rates. Western blotting was used to detect the levels of PI3K／Akt pathwayrelated proteins （p-Akt， Akt， p-mTOR and mTOR） 48 h after transfection. Immunofluorescence was used to evaluate the nuclear activity of NF-κB p65. Results  Oncomine bioinformatics analysis showed that the relative expression of S6K2 in cervical cancer tissues was significantly higher than that in normal cervical tissues in three cervical cancerrelated datasets （Biewenga, Scotto and Zhai）（P＜0.05）. The results of QPCR and Western blotting showed that the mRNA and protein levels of S6K2 after transfection of siRNA-S6K2 were significantly lower than those of the control group （P＜0.05）. The proliferative activity and levels of p-Akt and p-mTOR in HeLa and Caski cells transfected with siRNA-S6K2 decreased while the apoptotic rate increased， which was significantly different from that of cells transfected with siRNA-Ctrl （P＜0.05）. Immunofluorescence assay showed that the intranuclear fluorescence of cervical cancer cells transfected with siRNA-S6K2 decreased compared with the transfected siRNA-Ctrl cells, and the difference was statistically significant（P＜0.05）. Conclusion  S6K2 is highly expressed in cervical cancer tissues and participates in the proliferation and apoptosis of cervical cancer cells， so S6K2 has certain potential in the prevention and treatment of cervical cancer.

ResultsThe positive expression rates of Livin in colorectal cancer tissues， adjacent tissues and normal colorectal mucosa tissues were 64.0％（64/100）， 16.7％（5/30） and 0, respectively （P＜0.05）. Livin expression was correlated with TNM stage， invasion depth， lymph node metastasis, differentiation degree and preoperative CEA （P＜0.05）， but not with sex， age and tumor size（P＞0.05）. Cox multivariate regression analysis showed that Livin expression and TNM stage were independent prognostic factors for OS in stage Ⅰ-Ⅲ colorectal cancer（P＜0.05）. Conclusion  Livin may be associated with the occurrence， development and prognosis of colorectal cancer， and may provide important guidance for clinical diagnosis and evaluation of colorectal cancer.

Objective  To investigate the clinical value of circulating tumor cells （CTCs） in evaluating the prognostic value in patients with metastatic colorectal cancer undergoing microwave ablation. Methods  A total of 29 patients who received microwave ablation for hepatic metastases from colorectal cancer at the Second Hospital of Nanjing from January 2014 to May 2018 were enrolled in this study. Preoperative and postoperative blood samples from these patients were tested for CTCs. CTCs values and clinical data of the 29 cases were analyed. The data was analyzed by Chi square test or Fisher exact test, and the survival analysis was performed by Kaplan-Meier method and Log-rank test. Results  The short-term efficacy of microwave ablation for local lesions in 29 patients was assessed by imaging. The short-term outcome was complete ablation in 8 patients， partial ablation in 16 patients and progressive ablation in 5 patients. An increase in CTCs values after microwave ablation was found （Z=-2.489， P=0.013）. While， age， sex， synchronous colorectal liver metastasis or not， left or right colorectal cancer and the change of CTC values after microwave ablation were no statistically significant （P＞0.05）. The shortterm effect of local liver metastases in patients with CTCs ＜7/ml after microwave ablation was better than that in patients with CTCs ≥7/ml, and the difference was statistically significant （P=0.031）. The median overall survival（OS） of 29 patients was 30.0 months （95％CI： 10.7-49.3 months）. The median OS of patients under 60 years old were better than those aged 60 and above（P=0.008）. The median OS in patients with left colon cancer was better than that in patients with right colon cancer（P=0.028）. The patients with microwave ablation only have worse survival than combined treatment（P=0.001）. But gender， simultaneous or heterogeneous liver metastasis， elevated CTCs and postoperative CTCs／ml were not associated with median OS （P＞0.05）. Conclusion  Microwave ablation is an effective method for local treatment of colorectal cancer with liver metastases. Monitoring the number of CTCs before and after treatment may be helpful to evaluate the shortterm curative effects.

ObjectiveTo investigate the safety and effectiveness of enhanced recovery after surgery （ERAS） in perioperative period of colorectal cancer and the influence on postoperative nutritional status and immune.

Methods  A total of 61 patients with colorectal cancer from August 2015 to January 2017 were randomly divided into two groups. Thirty-one cases in fasttrack （FT） group were given perioperative management of ERAS and 30 cases in routine control （CT） group were given the conventional treatment. The albumin， prealbumin， transferrin， IgG， IgA， IgM， C-reactive protein， peripheral blood glucose， postoperative initial anal exhaust time， length of hospital stay， hospitalization cost and postoperative complications were compared between the two groups. Results  A total of 54 patients completed the experiment, with 27 patients in each group. Compared with CT group， the albumin， transferrin and IgG were significantly higher in FT group at 1， 3 and 5 days after surgery（P＜0.05）. The postoperative initial anal exhaust time and the length of hospital stay was shorter in FT group compared with CT group （P＜0.05）. The hospitalization costs of FT group and CT group was （20.3±1.2） and （27.6±0.9） thousand yuan， respectively （P＜0.05）. There was no significant difference in Creactive protein， peripheral blood glucose， IgA， IgM and post-operative complications between the two groups （P＞0.05）. Conclusion  ERAS is safe and effective for patients with colorectal cancer， and it has positive effect on postoperative nutritional status and immune of patients.