Objective The objective of this study was to investigate the role of sulindac in sensitization effects of NF-κB on apoptosis induced by TNF-α in human breast cancer.Methods The human breast cancer MCF-7 cell line was added sulindal in the logarithmic growth phase and the final concentrations of sulindac were 0.5 and 1.0 mmol/L.The cells in control group was cultured without adding succinic acid.After sulindac treatment for 48 h,flow cytometry,MTT and Western blotting were used to analyze the effect and mechanism of cell growth in MCF-7 cells.Results The inhibitory rate of cell proliferation was(29.17±1.23)% and(38.15±1.51)% in MCF-7 cells treated with 0.5 and 1.0 mmol/L of Sulindac for 48 h,respectively,when compared to the control group(1.15 ± 0.02)%(P<0.05).Compared with the control group,0.5 and 1.0 mmol/L of sulindac were significantly increased the G0/G1 phase in MCF-7 cells(P<0.05).The apoptosis rate of sulindac in MCF-7 cells was significantly higher than that in the control group(P<0.05).The expression levels of TNF-α were(2.09±0.67)% and(1.18±0.09)% in the concentrations of 0.5 and 1.0mmol/L sulindac,respectively,in MCF-7 cells when compared to the control group(7.42±0.56)%.Conclusion Sulindac has a certain effect on the growth of human breast cancer cells,which can promote the prolongation of cell cycle at the G0/G1 phase and improve sensitization of apoptosis.This mechanism may be related to the inhibition of TNF-α activity.

Objective To investigate the relationship between juxtapapillary duodenal diverticular (JPDD) and choledocholithiasis,and the effects of JPDD on endoscopic sphinctemtomy(EST) in treatment of choledocholithiasis.Methods Fifty-one cases of choledocholithiasis combined with JPDD (choledocholithiasis combined with JPDD group) and 210 cases of choledocholithiasis without JPDD (choledocholithiasis without JPDD group) were treated by EST,and the clinical data of patients in the two groups were retrospectively analyzed.The relationship between JPDD and choledocholithiasis was studied.The JPDD' influence on the intubation success rate of endoscopic retrograde cholangiopancreatography (ERCP),the success rate of the stone removal by EST and complication were analyzed.Results The incidences of choledocholithiasis in patients of JPDD diameter ＜ 1 cm,1-3 cm and ＞ 3 cm were 39.3％(11/28),53.2％ (33/62) and 7/8 respectively.The larger the JPDD diameter,the higher the incidence of choledocholithiasis,and there was statistical difference (P ＜ 0.01).The incidence of choledocholithiasis in peripheral type JPDD was significantly higher than that in parallel type and circumvolution type [81.0％(17/21) vs.41.9％(26/62) and 8/15],and there were statistical differences (P ＜0.05).There was no statistical difference in the intubation success rate of ERCP between the two groups (P ＞ 0.05),but the success rate of the stone removal by EST in choledocholithiasis combined with JPDD group was significantly lower than that in choledocholithiasis without JPDD group [91.8％ (45/49) vs.99.5％ (208/209)].The incidence of EST incision bleeding was significantly higher than that in choledocholithiasis without JPDD group [11.1％ (5/45) vs.1.9％ (4/208)],and there was statistical difference (P ＜ 0.01) ; there were statistical differences in the incidences of others complication between the two groups (P ＞ 0.05).Logistic regression analysis showed that JPDD was independent risk factor for EST incision bleeding (P =0.043).Conclusions JPDD is relative with choledocholithiasis.JPDD makes EST a little more difficult and risky,while EST is still a safe and effective therapy for choledocholithiasis patients combined with JPDD.

Objective:To investigate the correlation between microRNA-497 (miR-497) expression and invasion and metastasis and prognosis of colorectal cancer (CRC).Methods:The expression of miR-497 in 93 CRC patients (CRC group), 30 colorectal adenoma polyps (colorectal adenomatous polyp group) and 30 healthy patients (normal control group)were detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR); meanwhile, the content of serum carcinoembryonic antigen (CEA) was detected by chemiluminescence; After 3 years follow-up, the expression of miR-497 was measured again; the correlation between the expression of miR-497 and the clinicopathological characteristics and prognosis of CRC was analyzed.Results:The expression of miR-497 in CRC group was significantly lower than that at the end of postoperative treatment, adenomatous polyps group and normal control group ( P<0.01), and the content of CEA in CRC group was significantly higher than that at the end of postoperative treatment, adenomatous polyps group and normal control group ( P<0.01); At the end of postoperative treatment, the expression of serum miR-497 in CRC group was lower than that in normal control group ( P<0.05), and the content of serum CEA was higher than that in normal control group ( P<0.05). The diagnosis positive rate of serum miR497 expression in CRC group was significantly higher than that of serum CEA ( P<0.01), and they showed no correlation ( r=0.232, P>0.05). The pre-and post-operative serum miR-497 expression levels in the recurred and metastasis group were significantly lower than those in the no postoperative recurrence and metastasis group ( P<0.01). The preoperative expression of miR-497 was related to the differentiation degree of CRC, tumor node metastasis (TNM) stage, lymph node metastasis, distant metastasis and survival period ( P<0.01), but not with the age, sex, tumor size and location of the patients ( P>0.05). Cox multivariate analysis showed that tumor differentiation, TNM stage, preoperative miR-497 expression, lymph node metastasis and distant metastasis were all independent risk factors influencing prognosis of CRC patients ( P<0.05); The survival rate of 1, 2, 3 years in the miR-497 low expression group was lower than that in the miR-497 high expression group ( P<0.05). Conclusions:The preoperative low serum miR-497 expression levels are closely relative with invasion and metastasis and poor prognosis of CRC, and can be a prognostic indicator of CRC. The reduced postoperative serum miR-497 expression levels may be a predictor of the postoperative recurrence and metastasis of CRC.

Objective To investigate the relationship between quantitative detection of T lymphoma invasion and metastasis inducing factor (Tiaml) mRNA and invasion,metastasis and prognosis of patients with colorectal cancer.Methods The quantitative expression of Tiam1 mRNA in cancer tissues in colorectal cancers (n =97),in tissues of colorectal adenomatous polyposis (n =49) and normal control (n =30) were detected with real-time fluorescence quantitative-polymerase chain reaction (PCR).The relationship between the quantitative expression of Tiam1 mRNA in colorectal cancer tissues and the invasion,metastasis and the prognosis were analyzed.Results The levels of Tiam1 mRNA expression in the colorectal cancer group,were significantly higher than those in colorectal adenomatous polyposis group and normal controlgroup [(14.45±4.87)2-△△△Ct vs (3.77 ±3.15)2-△△Ct,(1.06 ±0.65)2-△△Ct,P ＜0.01].The levels of Tiam1 mRNA expression in the adenomatous polyposis with high grade intraepithelial neoplasia were signifiCantly higher than those in the adenomatous polyposis with low grade intraepithelial neoplasia [(5.26 ±2.45)2-△△Ct vs (3.78 ±2.19)2-△△Ct,P ＜0.01],which were both significantly higher than those in the adenomatous polyposis with no intraepithelial neoplasia [(2.13 ± 1.64)2-△△Ct,P ＜0.05 or P ＜ 0.01].The levels of Tiam1 mRNA expression in colorectal cancer group were positively related to the differentiated degree of tumor,depth of invasion,lymph node metastasis,tumor node metastasis (TNM) stages and the postoperative recurrence (P ＜ 0.01),and were negatively related to the postoperative survival time (P ＜0.01).Multivariate Cox regression analysis revealed that levels of Tiam1 mRNA expression in cancer tissues were independent prognostic factor for colorectal cancer (Exp =4.043).Conclusions The high Tiam1 mRNA expression levels in cancer tissues are closely related to invasion,metastasis and poor prognosis of colorectal cancer,and may be a prognostic indicator of colorectal cancer.The increase expression of Tiam1 mRNA in colorectal adenomatous polyposis may be related to colorectal tumorigenesis.

Objective To investigate the correlation between serum soluble epithelial cadherin (sE-cad) and postoperative recurrence and prognosis in patients with advanced gastric cancer.Methods The level of serum sE-cad in 85 patients with advanced gastric cancer (advanced gastric cancer group) was detected by ELISA technique preoperative and postoperative 1 month,and compared with 30 healthy controls(control group).The patients in advanced gastric cancer group were followed up for 3 years,the level of serum sE-cad in recurrent patients and non-recurrent patients was compared.Results The level of serum sE-cad in advanced gastric cancer group preoperative was significantly higher than that in control group [(24.3 ± 14.8) μ g/L vs.(9.4 ± 3.8) μ g/L,P ＜ 0.01].The level of serum sE-cad in advanced gastric cancer group was significantly decreased postoperative 1 month [(12.5 ± 6.4) μ g/L vs.(24.3 ± 14.8) μ g/L,P ＜0.01].The level of serum sE-cad in recurrent patients was significantly higher than that in non-recurrent patients and postoperative 1 month [(20.7 ±9.8)μg/L vs.(12.5 ±6.4),(14.8 ±6.2) μg/L,P＜0.01].Univariate analysis revealed that preoperative high serum sE-cad level was related with tumor size,differentiated degree,lymph node metastasis ratio,depth of tumor invasion (P ＜0.05),but had no relationship with histological type(P＞ 0.05).Elevated preoperative serum sE-cad level negatively affected the postoperative survival rate and recurrence rate.Multivariate Logistic regression anaiysis revealed that preoperative serum sE-cad level was an independent risk factor for postoperative 3 years survival rate in advanced gastric cancer (HR =2.068,P =0.013).Conclusions Preoperative elevated serum sE-cad level is related with pathologic features in patients with advanced gastric cancer,and may be an important prognostic factor.Postoperative monitoring the level of serum sE-cad is useful for evaluating the prognosis and recurrence.

Objective To investigate the correlation between expression of human telomerase reverse transcriptase (hTERT) mRNA in peripheral blood and postoperative recurrence and prognosis in the patients with advanced colorectal cancer (CRC).Methods The preoperative and 1 month postoperative quantitative expression of hTERT mRNA in peripheral blood in 85 patients with advanced CRC were detected with real-time fluorescence quantitative real-time fluorescence quantitative polymerase chain reaction (RTPCR).The quantitative expression of hTERT mRNA in peripheral blood in 30 normal controls were also detected.The postoperative prognosis was followed-up,and the quantitative expression of hTERT mRNA in peripheral blood in the patients with recurred CRC and the patients without recurred CRC were detected repeatedly.The factors that affected the preoperative hTERT mRNA expression were analyzed.The relationship was investigated between the hTERT mRNA expression levels and postoperative recurrence and prognosis in the patients with advanced CRC.Results The hTERT mRNA expression levels in peripheral blood in advanced CRC group were significantly higher than those in the normal controls [(6.22 ±5.85)2-△△Ct vs (0.45 ±0.27)2-△△Ct,(P ＜0.01)],and were significantly decreased at the time point of 1 month after the operation (P ＜ 0.01).The hTERT mRNA expression levels in the postoperative recurred CRC group were significantly higher than those in the 1 month postoperation group and no recurrence CRC group [(5.07 ± 2.87)2-△△Ct vs (1.83 ± 1.08)2-△△Ct,(2.15 ± 1.49)2-△△Ct,(P ＜0.01)].Univariate analysis revealed that preoperative high hTERT mRNA expression levels in peripheral blood were related to the differentiated degree of tumor,tumor size,lymph node metastasis ratio,and liver metastasis (P ＜ 0.05 or P ＜ 0.01).Multivariate logistic regression analysis revealed that differentiated degree of the tumor,lymph node metastasis,and liver metastasis were independent risk factors for the elevation of preoperative hTERT mRNA expression in the advanced CRC.Elevated preoperative hTERT mRNA expression was positively related to the postoperative recurrence rate,and was negatively related to postoperative survival rate.Conclusions Preoperative hTERT mRNA expression levels in peripheral blood were related to the important pathologic features in patients with advanced CRC,and might be a valuable prognostic factor.Postoperatively elevated hTERT mRNA expression in peripheral blood was related to recurrence of advanced CRC.

ObjectiveTo study the correlation between preoperative serum angiopoietin-2 (Ang2) levels and lymph node metastasis in early gastric cancer(EGC).MethodsPreoperative serum Ang-2 and CEA levels in 62 patients with EGG and 30 normal controls were measured by ELISA technique respectively.The metastasis of lymph node in these cases were determined by HE staining.The relation between preoperative serum Ang-2 levels and lymph node metastasis in EGC and pathologic characterization was investigated.ResultsCompared with normal controls,preoperative serum Ang-2 levels in EGC group were significantly elevated [( 282.5 ± 110.6 ) μg/L vs ( 187.4 ± 32.7 ) μg/L,P ＜ 0.01].Preoperative serum Ang-2 levels in node-positive gastric cancer group was significantly higher than that in the node-negative gastric cancer group [(34 7.2 ± 79.5 ) μg/L vs (265.6 ± 90.2) μg/L,P ＜ 0.01],while CEA levels changed a little.Preoperative serum Ang-2 levels in the EGC group presented marked correlation with the differentiated degree of the tumor tissue,tumor size,depth of tumor invasion but it had no correlation with the histological type.Preoperative serum Ang-2 levels were an independent high-risk factor of lymph node metastasis in EGC according to multivariate logistic regression analysis.ConclusionsPreoperative serum Ang-2 levels may be a good clinical predictor for lymph node metastasis of EGC,which is useful for the treatment direction.

Objective To investigate the correlation of the expression of human telomerase reverse transcriptase (hTERT) mRNA in peripheral and portal vein blood to liver metastasis and prognosis of patients with colorectal cancer (CRC).Methods From January 2009 to April 2011,a total of 181 patients diagnosed as primary CRC and received radical resection were enrolled.The relative quantitative expression of hTERT mRNA in preoperative peripheral blood and intraoperative portal vein blood were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR).All the patients were followed up for three years after operation.The relative quantitative expression of hTERT mRNA in peripheral blood before operation and intraoperative portal vein blood were compared between patients with synchronous liver metastasis (18 cases) and without synchronous liver metastasis (163 cases); between metachronous liver metastases (29 cases) and without metachronous liver metastases (152 cases).The relationship between the expression of hTERT mRNA in intraoperative portal vein blood and clinicopathological features of patients with CRC was analyzed.The t-test was used.Univariate and Multivariate Cox analysis were used for risk analysis of metachronous liver metastases.Log-rank test was used for comparisons of survival rate between the group with high hTERT mRNA expression in intraoperative portal vein blood during operation and the group with low expression.Results The relative quantitative expression of hTERT mRNA in preoperative peripheral blood and intraoperative portal vein blood in synchronous liver metastases group (8.04±3.79 and 11.88±4.19) was higher than that of no synchronous liver metastases group (4.30±2.81 and 4.94±3.37,t=5.159 and 8.084; both P＜0.01).The relative quantitative expression of hTERT mRNA in preoperative peripheral blood and intraoperative portal vein blood in metachronous liver metastases group (7.16±3.08 and 9.83 ± 2.96) was higher than that of no synchronous liver metastases group (4.11±2.58 and 4.56±3.09,t=5.648 and 8.467;both P＜0.01).The hTERT mRNA expression in intraoperative portal vein blood of patients with CRC changed significantly with the differentiated degree of tumor,tumor size,different invasion degree of tumor,lymph node metastasis,recurrence after operation,survival time (t =2.987,2.281,2.135,5.070,5.431 and 6.803,all P＜0.05).Univariate analysis revealed that the expression of hTERT mRNA in preoperative peripheral blood and intraoperative portal vein blood both were correlated with postoperative metachronous liver metastases (x2=9.522 and 16.393,both P＜0.01).Multivariate Cox analysis showed that both of them were independent risk factors (relative risk (RR)=4.286 and 9.783).The two-and three-year survival rates of the patients with high hTERT mRNA expression levels in intraoperative portal vein blood were 64.6 ％ and 52.3 ％,and those of low expression group were 91.4 ％ and 85.3 ％,and the differences between two groups in two-and three-year survival rate were statistically significant (x2 =5.313,P＜0.05; x2 =8.925,P＜0.01).Conclusions The expression of hTERT mRNA in intraoperative portal vein blood was closely correlated with the important pathologic features,prognosis and liver metastasis of patients with CRC and its predictive value of postoperative CRC metachronous liver metastases was higher than the expression of hTERT mRNA in preoperative peripheral blood.The hTERT mRNA in intraoperative portal vein blood can be a postoperative prognostic marker of patients with CRC.

Objective To study the difference and clinical significance of serum Angiopoietin-2 (Ang-2) level in gastric cancer, precancerous disease and the changes in post operation follow-up patients.MethodsSerum Ang-2 and CEA levels were detected with ELISA method in 158 gastric cancer patients, 31 chronic astrophic gastritis patients, 38 gastric adenomatous polyp patients, 28 chronic gastritis patients with benign gastric ulcer and 30 healthy controls.The changes of serum Ang2 levels in 131 patients with radical gastrectomy were measured in 2-year post-operational followed-up and the prognosis was also evaluated.The correlation between serum Ang-2 level and the pathologic character of gastric cancer was analyzed.ResultsThe serum Ang-2 and CEA levels of gastric cancer group were (331.8± 64.3) μg/L and (42.6 ± 37.3) μg/L, respectively, which were significantly higher than those of healthy control group [(187.4±32.7) μg/L and (4.2±3.1) μg/L]and benign gastric ulcer group [(197.3±35.4) μg/L and (4.5±3.2) μg/L](all P ＜0.01).The serum Ang-2 level in chronic astrophic gastritis group was also markedly increased (P＜0.05), however there was no significant change in CEA level.There was no obvious change of Ang-2 and CEA level in gastric adenomatous polyp group (P＞0.05).Pre-operation serum Ang-2 levels of the relapse group and the metastasis group were significantly higher than that of no relapse group (P＜0.05).Post-operation serum Ang-2 level of no relapse group decreased to normal range in 1 month and without obvious increase in 2 years follow-up.The serum Ang-2 level of the relapse group also decreased to normal range in 1 month post-operation, while which was significantly higher than that of no relapse group and normal control group from 6 months to 2 years after operation (P＜0.01).Serum Ang-2 level of the metastasis group was markedly higher than that of no relapse group and normal control group during 2 years after operation (P＜0.01).The 2-year survival rate of no relapse group was significant higher than that of relapse group and metastasis group.The change of serum Ang-2 level was correlated with in the gastric cancer differentiation degree, TNM pathologic classification, lymphatic metastasis, distant metastasis, the depth of invasion and the tumor size (P＜0.01), however not correlated with histopathology type and the location of tumor.ConclusionSerum Ang-2 level may be a new and valuable gastric cancer marker, which will help to monitor tumor relapse, migration and prognosis assessment.

In order to investigate the inhibitory effect of plasmid-mediated short hairpin RNA targeting vascular endothelial growth factor (VEGF) on the expression of VEGF mRNA in human gastric cancer cells, a plasmid vector for transcribing specific short hairpin RNA targeting VEGF (pU6-VEGF) was constructed, and then transfected into human gastric cancer cells using Lipofectamine2000. The VEGF mRNA expression level was detected by RT-PCR. RPMI1640 was used for blank control, and pSilencer 1.0-U6 empty plasmid for the negative control. Results showed the clone and sequence analysis revealed that the recombinant plasmid vector of pU6-VEGF was successfully constructed. The VEGF mRNA expression levels in blank control group, experimental group (pU6-VEGF) and negative control group (pSilencer1.0-U6) were 100%, 49% and 94%, respectively, indicating VEGF mRNA expression in the cells transfected with pU-VEGF vector was inhibited significantly as compared with blank control group and negative control group. It was concluded that the short hairpin RNA could significantly inhibit the expression of VEGF mRNA, which provided an experimental basis for treating human cancer with anti-angiogenesis.