Objective To investigate the clinical efficacy of the curettage and aspiration technique in laparoscopic radical gastrectomy for the treatment of gastric cancer.Methods The clinical data of 55 patients who received laparoscopic radical gastrectomy by curettage and aspiration technique with Peng's multifunctional operative dissector at the Shaoxing People's Hospital from June 2008 to February 2011 were retrospectively analyzed.Tumors located at the upper stomach in 10 patients,at the middle stomach in 15 patients and at the lower stomach in 30 patients.The numbers of patients had tumor in TNM stage Ⅰ,Ⅱ,Ⅲ A were 16,35 and 4.Patients were followed up via phone call and out-patient examination till October 2013.Results Laparoscopic radical gastrectomy was successfully carried out on all the 55 patients.Of the 55 patients,39 received laparoscopic distal subtotal gastrectomy and 16 received laparoscopic total gastrectomy.The operation time,volume of intraoperative blood loss,number of lymph nodes dissected,distances of proximal and distal resection margins to the tumors,time to flatus,time to fluid diet and duration of postoperative hospital stay and incidence of postoperative complications were (241 ± 42)minutes,(273±115)mL,32 ±9,(5.8±1.4)cm,(5.1 ±l.7)cm,(78 ±24)hours,(95 ±17)hours,(12 ±4)days and 7.3％ (4/55),respectively.Two patients were complicated with pulmonary infection,1 with anastomotic fistula,1 with incisional infection,and all of them were cured by symptomatic treatment.No patients died perioperatively.All the 55 patients were followed up for 12.0-55.0 months,and the mean time of follow-up was 35.9 months.The cumulative 48-month survival rate was 54.8％.The postoperative recurrence and metastasis rate was 10.9％ (6/55).Peritoneal metastasis was detected in 2 patients,liver metastasis in 1 patient,para-aortic nodes metastasis in 1 patient,residual gastric metastasis in 1 patient,and bone metastasis in 1 patient.Conclusion Laparoscopic radical gastrectomy by curettage and aspiration technique is safe and feasible,with the advantages of minimal trauma,low morbidity and quick recovery.

Objective To study the effect of transfection of antisense MBD1 gene eukaryotic expression vector on the expression of MBD1 gene in human cholangiocarcinoma cell line QBC-939.Methods The(constructed) antisense MBD1 gene eukaryotic expression vector was transfected into the human(cholangiocarcinoma) cell line QBC-939 using lipofectamine transfection reagents,and positive cell clones were obtained using G418 selection after transfection.The constructed recombinant vector was transfected into(QBC-939) cells successfully and was confirmed by amplifying the exogenous neo~R gene with PCR method.The expression level of MBD1 gene mRNA and protein was detected by RT-PCR and FCM methods respectively.Results Following the transfection,the MBD1 gene mRNA level in human cholangiocarcinoma cell line QBC-939 decreased from 0.912?0.022 to 0.215?0.017,and the MBD1 gene protein level also(decreased) from(80.19?5.05)% to(35.11?4.05)%.There were very significant differences on the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression vector transfection group(P

Objective To investigate the effects of the tumor suppressor gene PTEN in growing inhibition and down-regulating mTOR in cholangiocarcinoma QBC939 cells in vitro.Methods QBC939 cells were transfected with plasmids wild-type PTEN and C124S-PTEN in vitro.After transfection,the expression of the PTEN and phosphorylation of AKT and mTOR was detected by Western blot.Flow cytometry was used to analyze apoptosis and cell cycle of the transfected cells.Results Compared with the control,the expression of phosphorylation AKT was decreased and mTOR were down-regulated respectively when transfected with the wild-type PTEN.However,after transfection with mutation-type PTEN,the level of PTEN in the cells by increased,but phosphorylation AKT level and mTOR expression had no significant change.Conclusions PTEN can be actived by phosphorylated AKT.Actived AKT decreased the mTOR which led to tumor cells apoptosis and regulation of the tumor cell cycle.In the pathway of signal transmission of PI3K/AKT/PTEN/mTOR,PTEN and mTOR are closely related through phosphorylation of AKT.

Objective To investigate the relationship between clinical presentation and pathological characteristics in HBeAg negative chronic hepatitis B(CHB) patients with steatosis, and to find out the predictors of hepatic inflammation and fibrosis. Methods HgeAg negative CHB patients with (n=56) or without (n=60) steatosis confirmed clinically and pathologically were enrolled in the study. All patients were examined for fasting blood glucose(FBG), fasting insulin (FINS), triglyceride (TG), cholesterol (TC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyhransferase (GGT), alkaline phosphatase (ALP) albumin (Alb), globulin(Glb), homeostatic model assessment of insulin resistance (HOMA-IR), HBV-DNA and body mass index(BMI). The association of above parameters with hepatic inflammation, fibrosis and fatty deposition were analyzed statistically. Results It was demonstrated that BMI, FBG, FINS, TG, TC, GGT, ALP , Glb and HOMA-IR were significantly higher in HBeAg negative CHB patients with steatosis than those without steatosis (P<0.05). Whereas the levels of HBV-DNA, Alb, ALT and AST were significantly lower in HBeAg negative CHB patients with steatosis compared with those without steatosis (P<0.05). The hepatic inflammation and fibrosis were aggravated in patients with steatosis. It was implicated that BMI,FBG, FINS, TG, TC, GGT and HOMA-IR(all P values 0.05) were significant predictors for hepatic steatosis, while ALT, AST, Glb and HBV-DNA(all P values <0.05) were significant predictors for hepatic inflammation. And the predictors for hepatic fibrosis were ALT, AST, Alb, Glb and HBV-DNA(all P values <0.05). Conclusions Hepatic steatosis is common in HBeAg negative CHB patients which is positively associated with parameters including BMI, FBG, FINS, TG, TC, GGT, ALP and HOMA-IR. Besides steatosis, the hepatic inflammation and fibrosis are also aggravated in these patients.

Hypermethylation of the promoter region is one of the major mechanism of tumor suppressor gene inactivation. In order to provide a research tool for the study on the function of MBD1 gene in DNA methylation and tumorigenesis, antisense MBD1 gene eukaryotic expression plasmid was constructed and transfected into human biliary tract carcinoma cell line QBC-939 to observe its effect on the expression of MBD1 mRNA and protein by using RT-PCR and FCM respectively. Following the transfection, the mRNA level of MBD1 gene decreased from 0. 912 +/- 0.022 to 0.215 +/- 0. 017, and the protein level of MBD1 gene also decreased from (80.19 +/- 5.05) % to (35.11 +/- 4.05) %. There were very significant differences in the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression plasmid transfection group (P < 0.01). It was suggested that transfection with the antisense MBD1 gene eukaryotic expression plasmid can significantly reduce the expression level of MBD1 gene in QBC-939, and this study may provide a valid tool for the investigation of the function of MBD1 gene and its role in biliary tract carcinoma.
Biliary Tract Neoplasms/*metabolism ; Biliary Tract Neoplasms/pathology ; Cell Line, Tumor ; DNA Methylation ; DNA-Binding Proteins/*biosynthesis ; DNA-Binding Proteins/genetics ; Eukaryotic Cells/metabolism ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Oligonucleotides, Antisense/*genetics ; Plasmids/genetics ; Transcription Factors/*biosynthesis ; Transcription Factors/genetics ; Transfection

Hypermethylation in the promoter region is an important epigenetic mechanism for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferases are considered to be the main cause of promoter hypermethylation. In order to explore the roles of two methyltransferase members (DNMT1 and DNMT3b) in the cholangiocarcinoma tumorigenesis, antisense eukaryotic expression plasmid of DNMT1 and DNMT3b gene was constructed respectively, and were co-transfected into the human cholangiocarcinoma cell line QBC-939 to observe their biological effects on the cell growth and proliferation ability, apoptosis, cell cycle alteration, and the tumorigenesis ability in the subcutaneous tissue of nude mouse. The results demonstrated that co-transfection with antisense eukaryotic expression plasmid of DNMT1 and DNMT3b gene and single transfection with antisense eukaryotic expression plasmid of DNMT1 gene can suppress the growth and proliferation of QBC-939, block the cell cycle at G1 phase, increase the apoptosis rate, minimize the tumor size in the subcutaneous tissue of nude mouse. The suppressing biological effect of co-transfection is stronger than single transfection with antisense DNMT1. Meanwhile, single transfection with antisense eukaryotic expression plasmid of DNMT3b gene has no effects on the biological characteristics of QBC-939. This study suggests that DNMT1 gene plays a key role in DNA methylation and DNMT3b gene may act as an accessory to support its function in inactivation of tumor suppressor genes. Combination DNMT1 and DNMT3b will increase their biological effects and have the synergistic effect on suppressing the growth of human cholangiocarcinoma cell line QBC-939.
Apoptosis ; Biliary Tract Neoplasms/*metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cholangiocarcinoma/*metabolism ; DNA (Cytosine-5-)-Methyltransferase/*biosynthesis ; DNA (Cytosine-5-)-Methyltransferase/genetics ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Mice, Nude ; Neoplasm Transplantation

Objective To investigate the application value of time-spatial labeling inversion pulse (T-SLIP) in renal corticomedullary differentiaton and the best black blood inversion time (BBTI) value.Methods Totally 60 volunteers were included,who underwent abdominal MR scan with noncontrast-enhanced SSFP sequence combined with T-SLIP.All subjects were scanned with different BBTI (800,1 000,1 200,1 400,1 600,1 800 ms) using coronary T-SLIP SSFP sequence.The images quality was evaluated using a four-point scale method.The region of renal cortex and medulla was devised automatically based on the image training algorithm.The signal intensity ratio with the different BBTI was calculated through measuring the signal intensity of the renal cortex and medulla.And the best BBTI values were analyzed.Results When BBTI was 1 200 ms,the image score was the highest.The signal intensity ratio (SIR) had statistical difference among different BBTI groups (all P＜0.05),when BBTI was 1 200 ms,the SIR was the highest,and the contrast between the renal cortex and medulla was obvious.Conclusion T-SLIP technology can improve the visibility of renal corticomedullary without contrast agents.The optimal BBTI for the best corticomedullary differentiation is 1 200 ms.

Objective: To analyze the application of accelerated rehabilitation surgery in elderly patients with gastric cancer surgery and its influence on inflammation and nutritional indicators. Methods: From October 2017 to October 2018, 80 elderly patients with gastric cancer who underwent gastrectomy in Shaoxing People's Hospital were selected.According to random number table method, they were randomly divided into traditional control group and ERAS group, with 40 cases in each group.The traditional control group was treated by traditional perioperative treatment + operation, while ERAS group was treated with ERAS perioperative treatment + operation.The recovery and complications, inflammation and nutritional changes before operation, 1 day after operation and 3 days after operation, and the improvement of quality of life after operation were compared and analyzed between the two groups. Results: In the ERAS group, the first exhaust time[(2.3±0.8)d] and defecation time[(2.5±0.4)d]were shorter than those in the traditional control group[(3.5±0.5)d and (3.7±0.6)d], and the incidence rate of complications (7.5%) was lower than that in the traditional control group (35.0%), the differences were statistically significant (t=8.72, 10.52, χ²=9.04, all P<0.05). On the first day after operation, the serum levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6(IL-6) in the ERAS group were (28.3±4.9)μg/L and (23.1±5.8)μg/L, respectively, which were lower than those in the traditional control group[(39.8±6.7)μg/L and (34.5±8.1)μg/L](t=8.76, 7.24, all P<0.05). The serum TNF-α and IL-6 levels in the ERAS group on the 3rd day after operation were (18.4±6.2)μg/L and (18.3±3.2)μg/L, respectively, which were lower than those in the traditional control group[(26.2±5.1)μg/L and (26.5±4.8)μg/L](t=6.14, 8.99, all P<0.05). The serum levels of albumin (Alb) and prealbumin (PRE) in the ERAS group on the first day after operation were (31.1±0.7)g/L and (161.3±7.2) mg/L, respectively, which were lower than those in the traditional control group[(30.2±0.9)g/L and (154.3±4.8)mg/L](t=4.99, 5.12, all P<0.05). The serum levels of Alb and PRE in the ERAS group on the 3rd day after operation were (33.4±0.5)g/L and (172.1±5.0)mg/L, respectively, which were lower than those in the traditional control group[(31.9±0.8)g/L and (165.3±3.2)mg/L](t=10.06, 7.24, all P<0.05). The WHOQOL-BREF score of the ERAS group[(91.4±6.5)points]was higher than that of the traditional control group[(80.3±7.8)points](P<0.05). Conclusion Accelerated rehabilitation surgery has good effect for elderly gastric cancer surgery.It can maintain the inflammation and nutritional status of patients after operation.It is worthy of clinical reference.

Hypermethylation of the promoter region is one of the major mechanism of tumor suppressor gene inactivation. In order to provide a research tool for the study on the function of MBD1gene in DNA methylation and tumorigenesis, antisense MBD1 gene eukaryotic expression plasmid was constructed and transfected into human biliary tract carcinoma cell line QBC-939 to observe its effect on the expression of MBD1 mRNA and protein by using RT-PCR and FCM respectively. Following the transfection, the mRNA level of MBD1 gene decreased from 0. 912±0. 022 to 0. 215±0.017, and the protein level of MBD1 gene also decreased from (80.19±5.05) % to (35.11±4.05)%. There were very significant differences in the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukary otic expression plasmid transfection group (P＜0.01). It was suggested that transfection with the antisense MBD1 gene eukaryotic expression plasmid can significantly reduce the expression level of MBD1 gene in QBC-939, and this study may provide a valid tool for the investigation of the function of MBD1 gene and its role in biliary tract carcinoma.

Hypermethylation in the promoter region is an important epigenetic mechanism for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferases are considered to be the main cause of promoter hypermethylation. In order to explore the roles of two methyltransferase members (DNMT1 and DNMT3b) in the cholangiocarcinoma tumorigenesis, antisense eukaryotic expression plasmid of DNMT1 and DNMT3b gene was constructed respectively, and were co-transfected into the human cholangiocarcinoma cell line QBC-939 to observe their biological effects on the cell growth and proliferation ability, apoptosis, cell cycle alteration, and the tumorigenesis ability in the subcutaneous tissue of nude mouse. The results demonstrated that co-transfection with antisense eukaryotic expression plasmid of DNMT1 and DNMT3b gene and single transfection with antisense eukaryotic expression plasmid of DNMT1 gene can suppress the growth and proliferation of QBC-939, block the cell cycle at G1 phase, increase the apoptosis rate, minimize the tumor size in the subcutaneous tissue of nude mouse. The suppressing biological effect of co-transfection is stronger than single transfection with antisense DNMT1. Meanwhile, single transfection with antisense eukaryotic expression plasmid of DNMT3b gene has no effects on the biological characteristics of QBC-939. This study suggests that DNMT1 gene plays a key role in DNA methylation and DNMT3b gene may act as an accessory to support its function in inactivation of tumor suppressor genes. Combination DNMT1 and DNMT3b will inhere their biological effects and have the synergistic effect on suppressing the growth of human cholangiocarcinoma cell line QBC-939.