Objective To investigate the effects of microRNA(miRNA,miR)-887-3p on the proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells and its underlying molecular mechanism.Methods Annulus fibrosus tissues were obtained from 8-week-old SPF-grade SD male rats,centrifuged to prepare and identify annulus fibrosus cells.Rats in the experiment were randomly divided into four groups:a Normal group consisting of primary annulus fibrosus cells without any treatment;a Control group treated with 10 ng/mL interleukin-1β(IL-1β)for 24 hours to establish a degenerative cell model;an interference group(miR-887-3p inhibitor)transfected with miR-887-3p inhibitor using Lipo3000 based on the Control group;and an overexpression group(miR-887-3p mimics)transfected with miR-887-3p mimics using Lipo3000 based on the Control group.CCK-8 assay was used to assess cell viability;flow cytometry was used to measure cell apoptosis rates;real-time fluorescence quantitative PCR(qPCR)was used to detect the expression levels of miR-887-3p and murine double minute 4(MDM4)mRNA;Western blotting was used to measure the protein expression levels of MDM4,Bcl-2,and Caspase-3.Results Immunofluorescence staining of isolated and cultured cells revealed a Collagen I positive rate of over 90％in rat intervertebral disc annulus fibrosus cells,indicating a cell purity level greater than 90％.Real-time fluorescence qPCR results showed that after establishing an annulus fibrosus degenerative cell model using IL-1β,the expression level of miR-887-3p significantly increased compared to the Normal group(P＜0.001).Compared to the Control group,transfection with miR-887-3p inhibitor resulted in a significant decrease in its expression level(P＜0.001).The CCK-8 assay showed that compared to the Normal group,cell viability significantly decreased in the Control group(P＜0.001).Compared to the Control group,cell proliferation ability significantly increased after miR-887-3p inhibition,and significantly decreased after overexpression of miR-887-3p.Flow cytometry results revealed that compared to the Normal group,the apoptosis rate in the Control group significantly increased(P＜0.001).Compared to the Control group,the cell apoptosis rate significantly decreased in the miR-887-3p interference group(P＜0.001)and increased in the overexpression group(P＜0.001).Western blotting analysis showed that compared to the Normal group,Bcl-2 expression level significantly decreased(P＜0.001)and Caspase-3 expression level significantly increased(P＜0.001)in the Control group.Compared to the Control group,Bcl-2 and MDM4 expression levels significantly increased(P＜0.01),and Caspase-3 expression level significantly decreased(P＜0.01)in the miR-887-3p interference group;whereas in the overexpression group,Bcl-2 and MDM4 expression levels significantly decreased(P＜0.05),and Caspase-3 levels significantly increased(P＜0.05).Real-time fluorescence qPCR and protein immunoblotting results showed that after interfering with miR-887-3p,the expression of MDM4 protein and mRNA increased(P＜0.001);after overexpressing miR-887-3p,their expression decreased(protein,P＜0.01;mRNA,P＜0.001).Conclusion MiR-887-3p may modulate the cell proliferation and apoptosis of rat intervertebral disc annulus fibrosus cells by regulating MDM4 expression,thereby influencing the development and progression of disc degeneration.

Objective:To evaluate the long-term efficacy of laparoscopic-assisted natural orifice specimen extraction surgery (NOSES) colectomy using Cai tube for treating left-sided colorectal cancer.Methods:This was a randomized controlled trial. Inclusion criteria were as follows: preoperative pathological diagnosis of left-sided colorectal adenocarcinoma (rectal, sigmoid colon, descending colon, or left transverse colon cancer with the caudad margin ≥8 cm from the anal margin); preoperative abdominal and pelvic computed tomography (or magnetic resonance imaging) showing maximum tumor diameter <4.5 cm; and BMI <30 kg/m 2. Patients with synchronous multiple primary cancers or recurrent cancers, a history of neoadjuvant chemoradiotherapy, preoperative evidence of significant local infiltration, distant metastasis, or complications such as intestinal obstruction and intestinal perforation, or who were not otherwise considered suitable for laparoscopic surgery were excluded. A random number table was used to randomize sequential patients to NOSES surgery using Cai tube (non-assisted incision anal sleeve: patent number ZL201410168748.2) (NOSES group) or traditional laparoscopic-assisted surgery (CLS group). Relevant clinical data of the two groups of patients were analyzed, the main outcomes being disease-free survival, overall survival, overall recurrence rate, and local recurrence rate 5 years after surgery. Results:Patients in both study groups completed the surgery successfully with no requirement for additional surgery. After mean 70 (7–83) months postoperative follow-up, the 5-year overall postoperative survival in the NOSES and CLS groups was 90.0% and 83.3%, respectively ( P=0.455); disease free survival was 90.0% and 83.3%, respectively ( P=0.455); overall recurrence rate 6.6% and 10.0%, respectively ( P=0.625); and local recurrence rate both were 3.3% ( P=0.990), respectively. None of these differences was statistically significant. Conclusions:NOSES and CLS have similar long-term efficacy, and NOSES deserves to be used in clinical practice.

Objective:To evaluate the long-term efficacy of laparoscopic-assisted natural orifice specimen extraction surgery (NOSES) colectomy using Cai tube for treating left-sided colorectal cancer.Methods:This was a randomized controlled trial. Inclusion criteria were as follows: preoperative pathological diagnosis of left-sided colorectal adenocarcinoma (rectal, sigmoid colon, descending colon, or left transverse colon cancer with the caudad margin ≥8 cm from the anal margin); preoperative abdominal and pelvic computed tomography (or magnetic resonance imaging) showing maximum tumor diameter <4.5 cm; and BMI <30 kg/m 2. Patients with synchronous multiple primary cancers or recurrent cancers, a history of neoadjuvant chemoradiotherapy, preoperative evidence of significant local infiltration, distant metastasis, or complications such as intestinal obstruction and intestinal perforation, or who were not otherwise considered suitable for laparoscopic surgery were excluded. A random number table was used to randomize sequential patients to NOSES surgery using Cai tube (non-assisted incision anal sleeve: patent number ZL201410168748.2) (NOSES group) or traditional laparoscopic-assisted surgery (CLS group). Relevant clinical data of the two groups of patients were analyzed, the main outcomes being disease-free survival, overall survival, overall recurrence rate, and local recurrence rate 5 years after surgery. Results:Patients in both study groups completed the surgery successfully with no requirement for additional surgery. After mean 70 (7–83) months postoperative follow-up, the 5-year overall postoperative survival in the NOSES and CLS groups was 90.0% and 83.3%, respectively ( P=0.455); disease free survival was 90.0% and 83.3%, respectively ( P=0.455); overall recurrence rate 6.6% and 10.0%, respectively ( P=0.625); and local recurrence rate both were 3.3% ( P=0.990), respectively. None of these differences was statistically significant. Conclusions:NOSES and CLS have similar long-term efficacy, and NOSES deserves to be used in clinical practice.

OBJECTIVETo investigate the feasibility and safety of membrane-based right-sided approach of laparoscopic suprapancreatic lymph node dissection for advanced distal gastric cancer.

METHODSThe clinical data of 41 patients with advanced distal gastric cancer who underwent laparoscopic gastrectomy using membrane-based right-sided approach for laparoscopic suprapancreatic lymph node dissection at the Department of Gastrointestinal Surgery, Zhongshan Hospital of Xiamen University from January 2016 to January 2018 were retrospectively analyzed. There were 24 males and 17 females with a mean age of 56.8 years and a mean body mass index of 22.6 kg/m². Membrane-based right-sided approach of laparoscopic suprapancreatic lymph node dissection contained 4 steps briefly: (1) dissection of mesenteria above the head of pancreas: the tri-junction of pancreas-duodenum was cut to expose and identify the logo of Benz;clearance of the membrane of No.5a was performed towards the left, and then expanded to the posterior layer of No.12a. (2) dissection of the V shape dorsal mesogastrium: membrane bridge at splenic artery trunk root was cut; in suprapancreatic space, clearance was performed towards to the left to the middle of the splenic artery trunk and expanded to the posterior Tolds plane upwards to the posterior phrenic angle and retroperitoneal esophagus, then the surrounding tissue of anterior abdominal aorta. (3) dissection of the U shape mesenteria:membrane bridge at common hepatic artery root was cut; mesentery was separated; the left gastric vein was freed and ligated at its root; in posterior pancreatic space, the mesentery of No.7, No.9 and No.8 was dissected in turns; the left gastric artery was high ligated and cut; the portal vein and posterior dorsal mesogastrium Toldt plane was routinely exposed; clearance was performed to right for No.8a and upward to the hepatic portal meeting at posterior mesentery No.12 plane. (4) dissection of the upper triangular area of pylorus: the trigone mesentery was cut along the upper edge of the pylorus; No.12a was swept upward along the gastric ventral mesentery; the upper boundary(No.8a) on the right side of the U-shaped membrane was joined. Intraoperative and postoperative presentations were analyzed.

RESULTSLaparoscopic gastrectomy for advanced distal gastric cancer with membrane-based right-sided approach of laparoscopic suprapancreatic lymph node dissection was successfully carried out in all the 41 patients. Distal gastric mesenteria en bloc resection was successfully performed. The operation time was (145.2±25.4) minutes and intraoperative blood loss was (53.3±18.3) ml without massive bleeding and severe complication. Number of lymph nodes dissected was 41.1±6.4, and number of suprapancreatic lymph node dissected was 23.3±3.7 without residual cancer at cut margin by pathology. Postoperative drainage volume was (65.8±21.7) ml; time to withdraw of catheter was (7.0±1.7) days; time to fluid intake was (3.5±1.8) days; postoperative hospital stay was (10.4±2.8) days; time to postoperative anal exhaust was (3.3±1.1) days. No complications, such as chyle leakage, postoperative massive bleeding, anastomotic leakage, abdominal cavity infection or gastroplegia occurred within 30 days after surgery.

CONCLUSIONMembrane-based right-sided approach of laparoscopic suprapancreatic lymph node dissection for advanced distal gastric cancer can achieve en bloc resection and conform to the radical principle of oncology, and is safe and feasible.

OBJECTIVETo explore the feasibility of laparoscopic-assisted natural orifice specimen extraction radical left colectomy.

METHODSRetrospective analysis was performed on clinicopathological dada of 15 colorectal patients who were treated by laparoscopic-assisted anal specimen extraction radical left colectomy with self-developed surgical instrument Cai tube between January and September in 2014. Tumor location included descending colon (n=3), the junction of descending colon and sigmoid colon (n=2), the sigmoid colon (n=6) and upper rectum (n=4). Clinical efficacy of patients was observed.

RESULTSThere were no perioperative deaths or postoperative complications, such as anastomotic bleeding or leakage. The median operation time was 257 (range 103-337) min, median blood loss was 50(range 20-200) ml, median time to first flatus was 3 (range 1-5) d and median hospital stay was 14 (range 11-21) d. All the patients had good quality of life and normal defecation function without tumor recurrence or metastasis after 1-8 months of follow-up.

CONCLUSIONLaparoscopic-assisted anal specimen extraction radical left colectomy is safe and feasible.

Colectomy ; Colon, Sigmoid ; Humans ; Laparoscopy ; Length of Stay ; Operative Time ; Postoperative Complications ; Quality of Life ; Rectum ; Retrospective Studies

Objective To investigate the expression of ASK1 and PRKCD in the process of monocyte differentiation, and explore their role in functional changes of hypersplenism spleen macrophages (MΦ) in portal hypertension (PH). Methods U937 cells were stimulated to differentiate into monocyte/macrophage-like cells by cultivation in PMA and the mRNA expressions of ASK1 and PRKCD were detected by q-PCR and the changes of protein expression were identified by western blot analysis. The secretion of phagocytose related cytokines such as IL-10 and TNF-a were tested by ELISA, and the function of the macrophage-like cells were studied by chicken red blood cell phagocytose test. Results The expressions of PRKCD and ASK1 mRNA were gradually decreased along with the cell differentiation, while the secretion of TNF-αwas increased, IL-10 secretion reached a maximum at 24 h after PAM stimulation, and then gradually fell. The expression of ASK1 and p-ASK1 were rapidly increased compared with the non-stimulated U937 cells, while the expression of PRKCD and p-PRKCD were sightly declined. The phagocytose test show that U937 cells induced with PMA were able to swallow the chicken red blood cell.Conclusion Up-regulated protein expression of ASK1 and p-ASK1 and down-regulated protein expression of PRKCD and p-PRKCD in the process of PMA induced monocyte differentiation, are consist with the expression changes of splenic macrophage phagocytosis in hypersplenism, which leads to increased activity of MΦ.

Objective To explore the feasibility of laparoscopic-assisted natural orifice specimen extraction radical left colectomy. Methods Retrospective analysis was performed on clinicopathological dada of 15 colorectal patients who were treated by laparoscopic-assisted anal specimen extraction radical left colectomy with self-developed surgical instrument Cai tube between January and September in 2014. Tumor location included descending colon (n=3), the junction of descending colon and sigmoid colon (n=2), the sigmoid colon (n=6) and upper rectum (n=4). Clinical efficacy of patients was observed. Results There were no perioperative deaths or postoperative complications , such as anastomotic bleeding or leakage. The median operation time was 257 (range 103-337) min, median blood loss was 50(range 20-200) ml, median time to first flatus was 3(range 1-5) d and median hospital stay was 14 (range 11-21) d. All the patients had good quality of life and normal defecation function without tumor recurrence or metastasis after 1-8 months of follow-up. Conclusion Laparoscopic-assisted anal specimen extraction radical left colectomy is safe and feasible.

Objective To investigate the expression of ASK1 and PRKCD in the process of monocyte differentiation, and explore their role in functional changes of hypersplenism spleen macrophages (MΦ) in portal hypertension (PH). Methods U937 cells were stimulated to differentiate into monocyte/macrophage-like cells by cultivation in PMA and the mRNA expressions of ASK1 and PRKCD were detected by q-PCR and the changes of protein expression were identified by western blot analysis. The secretion of phagocytose related cytokines such as IL-10 and TNF-a were tested by ELISA, and the function of the macrophage-like cells were studied by chicken red blood cell phagocytose test. Results The expressions of PRKCD and ASK1 mRNA were gradually decreased along with the cell differentiation, while the secretion of TNF-αwas increased, IL-10 secretion reached a maximum at 24 h after PAM stimulation, and then gradually fell. The expression of ASK1 and p-ASK1 were rapidly increased compared with the non-stimulated U937 cells, while the expression of PRKCD and p-PRKCD were sightly declined. The phagocytose test show that U937 cells induced with PMA were able to swallow the chicken red blood cell.Conclusion Up-regulated protein expression of ASK1 and p-ASK1 and down-regulated protein expression of PRKCD and p-PRKCD in the process of PMA induced monocyte differentiation, are consist with the expression changes of splenic macrophage phagocytosis in hypersplenism, which leads to increased activity of MΦ.

Objective To explore the feasibility of laparoscopic-assisted natural orifice specimen extraction radical left colectomy. Methods Retrospective analysis was performed on clinicopathological dada of 15 colorectal patients who were treated by laparoscopic-assisted anal specimen extraction radical left colectomy with self-developed surgical instrument Cai tube between January and September in 2014. Tumor location included descending colon (n=3), the junction of descending colon and sigmoid colon (n=2), the sigmoid colon (n=6) and upper rectum (n=4). Clinical efficacy of patients was observed. Results There were no perioperative deaths or postoperative complications , such as anastomotic bleeding or leakage. The median operation time was 257 (range 103-337) min, median blood loss was 50(range 20-200) ml, median time to first flatus was 3(range 1-5) d and median hospital stay was 14 (range 11-21) d. All the patients had good quality of life and normal defecation function without tumor recurrence or metastasis after 1-8 months of follow-up. Conclusion Laparoscopic-assisted anal specimen extraction radical left colectomy is safe and feasible.

OBJECTIVETo investigate the expressions of extracellular signal-regulated kinase (ERK) and the BH3-only subgroup of Bcl-2 related proteins (Bim) in multidrug-resistant hepatocellular carcinoma (HCC) cells and their association with drug resistance in the cells.

METHODSThe multdrug-resistant HepG-2 cell line was established by treatment with gradually increasing doses of ADM. CCK-8 assay was used to determine the drug sensitivity of the cells, and the expressions of MRP-1, P-gp, ERK1, ERK2, ERK5, and Bim were detected with Western blot. Bim mRNA expression level was measured using quantitative real-time PCR.

RESULTSThe drug resistance indices to ADM, 5-FU and CDDP was 6.8, 4.10, and 4.5 in HepG-2/ADM cells, respectively. The drug-resistantcells showed marked up-regulation of MRP-1, P-gp, ERK1, ERK2 and ERK5 with down-regulated phosphorylated ERK2 protein expression but no significant changes in phosphorylated ERK1 protein expression to result in a decreased ratio of P-ERK1/2 and P-ERK1/2. Bim mRNA and protein expressions were both decreased in HepG-2/ADM cells.

CONCLUSIONERK and Bim are related to multidrug resistance in HepG-2/ADM cells.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; Down-Regulation ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; Phosphorylation ; Up-Regulation