miR-497 inhibits the growth and metastasis of SGC-7901 human gastric cancer anoikis resistant cells via blocking Wnt/β-catenin signaling pathway / 细胞与分子免疫学杂志

Objective To investigate the effects of microRNA497 (miR-497) on the metastasis of gastric cancer and its possible molecular mechanism. Methods SGC-7901 gastric cancer parent cells were cultured in an ultra-low adhesion environment, and the anoikis resistance model of SGC-7901 cells was created after re-adhesion. Clone formation assay, flow cytometry, TranswellTM test and scratch healing test were used to detect the differences of biological behavior compared with their parent cells. Fluorescence quantitative PCR was performed to detect the expression of miR-497. Western blot analysis was used to detect the changes of key proteins of Wnt/β-catenin signaling pathway and epithelial mesenchymal transformation (EMT) related proteins such as vimentin and E-cadherin. Parent cells and anoikis resistant SGC-7901 cells were transfected with miR-497 inhibitor or miR-497 mimic, and CCK-8 assay was used to detect the proliferation activity. TranswellTM invasion assay was performed to detect the invasion ability of cells. TranswellTM migration test and scratch healing assay was used to determine the migration ability. Western blot analysis was used to detect the expressions of Wnt1, β-catenin, vimentin and E-cadherin. By transfecting miR-497 mimic into the anoikis resistance SGC-7901 cells and inoculating them subcutaneously in nude mice, the changes in the volume and mass of tumor tissues were measured and recorded. Western blot analysis was used to determine the expressions of Wnt1, β-catenin, vimentin and E-cadherin of tumor tissues. Results Compared with the parent cells, the anoikis resistance SGC-7901 gastric cancer cells had faster proliferation rate, stronger colony formation, lower apoptosis rate, stronger invasion and migration ability. The expression of miR-497 was significantly decreased. After down-regulation of miR-497, the proliferation ability, invasion and migration ability were significantly enhanced. The expressions of Wnt1, β-catenin and vimentin increased significantly, while E-cadherin decreased notably. The results of up-regulation miR-497 were the opposite. The tumor growth rate, tumor volume and mass of miR-497 overexpression group were significantly lower than those of control group. The expressions of Wnt1, β-catenin and vimentin decreased significantly, while the expression of E-cadherin increased significantly. Conclusion The expression of miR-497 is low in the anoikis resistance SGC-7901 cells. miR-497 can inhibit the growth and metastasis of gastric cancer cells by blocking Wnt/β-catenin signaling pathway and EMT.

Effect of perioperative continuous use of aspirin on bleeding in laparoscopic anterior resection for rectal cancer: a prospective study / 中华消化外科杂志

Objective To explore the effect of perioperative continuous use of aspirin on bleeding in laparoscopic anterior resection for rectal cancer (RC) in patients taking low dose aspirin.Methods The prospective study was conducted.The clinicopathological data of 96 RC patients taking low dose aspirin who were admitted to the Shengjing Hospital of China Medical University from September 2014 to September 2016 were collected.All the 96 patients were divided into the aspirin group (perioperative continuous use of aspirin) and non-aspirin group (discontinuation of aspirin at 7 days preoperatively and taking aspirin at 3 days postoperatively)by random number table.Laparoscopic anterior resection for RC was applied to patients by the same team of doctors.Observation indicators:(1) comparison of surgical and postoperative situations between the 2 groups;(2) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect the postoperative survival of patients up to January 2017.Measurement data with normal distribution were represented as (x)±s.Comparisons between groups were evaluated with the t test.Comparisons of count data were analyzed using the chi-square test and Fisher exact probability.Comparison of ordinal data was analyzed using the nonparametric test.Results All the 96 patients were enrolled into the study,including 50 in the aspirin group and 46 in the non-aspirin group.(1) Comparison of surgical and postoperative situations between the 2 groups:96patients underwent successful laparoscopic anterior resection for RC,including 1 with conversion to open surgery and 95 undergoing laparoscopic anterior resection for RC,without perioperative death,postoperative blood transfusion and rcoperation.Cases with conversion to open surgery,operation time,volume of intraoperative blood loss,decreasing value of postoperative hemoglobin (Hb),time to anal exsufflation,peritoneal drainage volume from 1-3 days postoperatively,cases with postoperative hematoehezia,cardio-cerebrovascular complications and overall complications (including postoperative hematochezia and cardio-cerebrovascular complications),duration of hospital stay,hospital expenses,cases in stage Ⅰ,Ⅱ and Ⅲ of postoperative TNM stage,postoperative coagulation indexes of platelet,prothrombin time,international normalized ratio,prothrombin activity,activated partial thromboplastin time,thrombin time and fibrinogen were 0,(112± 18) minutes,(39± 18) mL,(4.3±2.8)g/L,(57±24)hours,(22±9)mL/d,6,0,10,(8.6±2.5)days,(6 739±481)yuan,11,35,4,(236±80) × 109/L,(12.7± 1.1) seconds,1.00±0.08,101％ ± 15％,(28 ±4) seconds,(15.5 ± 1.9) seconds,(3.2±1.0)g/L in the aspirin group and 1,(118±16)minutes,(38±22)mL,(3.5±3.0)g/L,(55±29)hours,(20±8)mL/d,4,1,8,(9.1±2.3)days,(6606±510)yuan,5,36,5,(211±49)×109/L,(12.5±0.7)seconds,1.00±0.06,103％±11％,(29±3)seconds,(15.3±1.7)seconds,(3.1±0.7)g/L in the non-aspirin group,respectively,with no statistically significant difference between the 2 groups (t =-1.737,0.204,1.416,0.380,1.365,x2=0.038,0.107,t=-1.082,1.322,Z=-1.370,t=1.850,0.978,0.872,-0.712,-1.291,0.311,0.585,P＞0.05).Ten patients with postoperative hematochezia in the 2 groups were cured,without special treatment.One patient in the non-aspirin group was complicated with deep venous thrombosis and then was cured by single anticoagulant drug.Two and 2 patients in the aspirin group were respectively complicated with urinary retention and urinary tract infection.One,1 and 1 patients in the non-aspirin group were respectively complicated with inflammatory intestinal obstruction,urinary retention and urinary tract infection,and then were cured by conservative treatment.(2) Follow-up situations:of 96 patients,95 were followed-up for 4-27 months,with a median time of 13 months.During the follow-up,3 patients died and 92 had survival.Conclusion The perioperative continuous use of aspirin cannot increase risk of bleeding in laparoscopic anterior resection for RC in patients taking low dose aspirin.

Application of transanal specimen extraction via prolapsing approach in laparoscopic anterior resection for ultra-low rectal cancer / 中华消化外科杂志

Objective To explore the application value and clinical efficacy of the transanal specimen extraction via prolapsing approach in laparoscopic anterior resection for ultra-low rectal cancer.Methods The clinical data of 27 patients with ultra-low rectal cancer who underwent transanal specimen extraction via prolapsing approach in laparoscopic anterior resection for ultra-low rectal cancer at the Shengjing Hospital of China Medical University from July 2010 to July 2013 were analyzed retrospectively.The average operation time, average volume of intraoperative blood loss, average number of lymph nodes dissection, average distance to resection margin, average length of resected specimen, results of postoperative pathological examination, time for postoperative outoff-bed activity, time to anal exsufflation, gastric tube removal time and postoperative complications were recorded.The visual analogue scale (VSA) pain score and Wexner score for evaluating fecal incontinence were performed at postoperative week 1 and at postoperative month 1, respectively.The anal function was tested at postoperative month 3 and 12.The follow-up including tumor metastasis and recurrence, Wexner score and anorectal anometry was performed by outpatient examination and telephone interview once every 3 months for 2 years after operation and then once every year up to October 2014.Measurement data with normal distribution was presented as-x ± s and average (range).Repeated measures data were analyzed by the repeated measures ANOVA.Results All the patients received successful operations without procedure change or intraoperative accident.The average operating time, average volume of intraoperative blood loss, average number of lymph nodes dissection, average distance to distal resection margin and average length of resected specimen were 140 minutes (range, 115-173 mintues), 27 mL(range, 15-55 mL), 17(range, 14-20), 1.7 cm(range, 1.3-2.2 cm) and 18.5 cm(range, 14.7-22.4 cm), respectively.Postoperative TNM stages: T2N0M0 was detected in 19 patients, T2N1M0 in 3 patients,T3N0M0 in 4 patients and T3N1M0 in 1 patients.The time for postoperative out-off-bed activity and time to anal exsufflation were 8.8 hours (range, 7.0-13.0 hours) and 51 hours (range, 30-79 hours).Twenty-seven patients had the gastric tube removal after operation with fluid diet intake at postoperative hour 24 and semi-fluid diet intake at postoperative hour 48.One male patient was complicated with urinary retention at postoperative day 3 and 1 with anastomotic leakage at postoperative day 9, they were cured by symptomatic treatment.VSA pain scores in all patients from 1 day to 6 days postoperatively were 5.6, 4.6, 4.0, 3.2, 2.2 and 1.3.The average duration of hospital stay was 11.1 days (range, 7.0-19.0 days).Patients had good healing of abdominal incision at postoperative week 2.All the patients were followed up for a average time of 24.8 months (range, 15.0-32.0 months) without tumor metastasis and recurrence.Wexner score was 2.6 (range, 1.0-4.0) at postoperative month 1.The results of anorectal anometry: maximum anorectal resting pressure (MARP) and maximum anorectal systolic pressure were (267 ±23)mmHg (1 mmHg =0.133 kPa) and (305 ± 23)mmHg before operation, (266 ± 40)mmHg and (300 ± 38)mmHg at postoperative month 3, (267 ± 33)mmHg and (315 ± 30)mmHg at postoperative month 12, respectively, with no significant difference in the changing trend between pre-and post-operation (F =0.510, 1.390, P ＞ 0.05).Anorectal resting vector volume and anorectal systolic vector volume were (45 594 ± 1 981) cm (mmHg) 2 and (98 480 ± 8 165) cm (mmHg) 2 before operation, (40 310 ±3 465)cm(mmHg)2 and (78 461 ±6 777)cm(mmHg)2 at postoperative month 3, (40 385 ± 3 379) cm(mmHg) 2 and (82 082 ± 10 383) cm(mmHg) 2 at postoperative month 12, respectively, with significant differences in the changing trend between pre-and post-operation (F =26.845, 48.090, P ＜ 0.05).Conclusion Transanal specimen extraction via prolapsing approach in laparoscopic anterior resection for ultra-low rectal cancer is safe, aesthetic and minimally invasive compared with the traditional laparoscopic surgery, with the advantages of radical cure of tumor and protection of anal function.

Laparoscopy-guided intersphincteric resection for low rectal stro-mal tumor / 中国肿瘤临床

Objective:To investigate the clinical security and feasibility of neoadjuvant chemotherapy with imatinib following lap-aroscopy-guided intersphincteric resection for patients with gastrointestinal stromal tumor of the low rectum (GSTLR). Methods:Clini-cal data of nine patients with GSTLR who were admitted to the Shengjing Hospital between January 2007 and January 2011 were re-viewed. These patients were treated with neoadjuvant imatinib chemotherapy after laparoscopic intersphincteric resection. Results:Pri-or to neoadjuvant chemotherapy, the tumor diameter ranged between from 5 cm to 9 cm (median=7.0 cm). After imatinib chemothera-py, the tumor diameter decreased to 2-4.5 cm (median=3.5 cm, P<0.001). Laparoscopic surgery through intersphincteric resection was performed after imatinib treatment for 3-24 months (median=7 months). All patients received a protective stoma, which was closed 3 months after the surgery. The Wexner scale scores ranged from 1 and 4 (median=2) prior to neoadjuvant imatinib chemotherapy and changed to 1-5 (median=2) after the chemotherapy (P=0.397). After stomal closure operation, the scores significantly increased to 4-9 (median=7, P<0.001) but were not statistically significantly different from those before the therapy. One year after laparoscopic surgery, the Wexner scale scores ranged from 1 to 5 (median=2, P=0.842). Six patients were treated with imatinib for 24 and 30 months after lap-aroscopic surgery. Recurrence in pelvis occurred in only one patient, who ceased imatinib administration at the 30th month after the sur-gery. Conclusions: Laparoscopic surgery through intersphincteric resection was secure and feasible and thus could be used for treat-ment of GSTLR.

Influence of RNA interference on MMP-9 gene expression in THP-1 cell line / 中国免疫学杂志

To study the inhibitory effect of RNA interference (RNAi) on MMP-9 gene expression in THP-1 cell line.To investigate the application of RNAi on the therapy of leukemia.Methods:Small interfering RNA ( siRNA) for MMP-9 gene was designed and transfected into THP-1 cells.MMP-9 mRNA expression was assessed by RT-PCR, and MMP-9 protein expression was tested by Western blot.MTT and trypan blue staining were used to observe the effect on the proliferation of THP-1 cells after RNAi.The changes in cell morphology were observed under the microscope.Results:The expressions of MMP-9 mRNA and protein were inhibited in THP-1 MMP-9 siRNA-transfected cells ,significantly lower than those of control cells.The results of MTT and trypan blue staining in-dicated that the proliferation ability of THP-1 cells obviously decreased after siRNA-transfected 48h and 72h.The growth of cells was in-hibited and the cells survival rate was significantly lower than that of control group ( P<0.05 ).The cells of control groups grew semi-quote wall under inverted microscope.The outline of cells was clear and the shape was uniform.The cells grew vigorously.While the growth of cells in siRNA group was inhibited.The morphology of siRNA group cells changed obviously by the Wright staining.Most cells expressed changes of apoptosis.Conclusion: siRNA for MMP-9 gene can not only reduce the expressions of MMP-9 mRNA and protein,but also inhibit the proliferation and induce apoptosis of THP-1 cells.

The effects of recombinant human growth hormone on the serum levels of cytokines severely burned patients / 中国基层医药

Objective To explore the effects of recombinant human growth hormone(rhGH) on the serum levels of cytokines in severely burned patients.Methods Thirty-six burn patients were en-rolled in the study and were randomly divided into 3 groups according to the rhGH dosage used,i.e. small(0.3IU?kg -1 ?d -1 ,A),large(0.6IU?kg -1 ?d -1 ,B) dose groups and control group(C,with normalsaline).The rhGH was administered beginning from 3 postburn days(PBDs) and lasted for 20 days.The dynamic changes in the serum levels of TNF-?,IL-6,IL-8 and LPS at different time points were observed.Results When compared with these in C group,the serum levels of TNF-?,IL-6 in A,B groups were decreased,especially in B group with earlier decrease and bigger range(P0.05).Conclusions rhGH might decrease the production of postburn inflammatory mediators.The clinical application of rhGH might be a supplementary measure in preventing and ameliorating postburn SIRS and MODS in severely burned patients.