Objective To study the mechanism of p55 inducing cell apoptosis, the 180 bp fragment of Puma promoter was cloned into the pGL3-basic luciferase reporter vector. The biological activity of Pumareporter plasmid was verified by cell transfection. Methods The target fragments of Puma were amplified by RT-PCR method and the fragments were inserted into the pGL3-basic luciferase reporter vector. The acquired Puma-Luc plasmid was transfected into H1299 cell line and detected its activity. Results Sequencing indicated that the amplified Puma promoter is correct. Dual-luciferase Reporter Assay showed the Puma-Luc constructs have promoter activity. Conclusion The cloning of human Puma gene promoter and the construction of its reporter vector were successful. This study will lay the foundation for further research on the function of p53 inducing apoptosis through mitochondrial pathway.

Objective Construct the eukaryotic expression vector of inhibitory member of the ASPP family (iASPP) and trans-fect it into colon carcinoma cell lines SW480 and Lovo by liposome .Then observe the expression of iASPP and detect the cell apop-tosis by flow cytometry .Methods The amplified PCR product was digested and inserted into pMD19-T simple vector and sub-cloned into eukaryotic expression vector pcDNA3 .1(+ ) .The recombinant eukaryotic expression plasmid pcDNA3 .1(+ )-iASPP was transfected into colon carcinoma cell lines SW480 and Lovo by liposome ,the iASPP expression was analyzed by RT-PCR .The cell apoptosis was detected by FCM .Results The eukaryotic expression plasmid pcDNA3 .1(+ )-iASPP was constructed success-fully ,the gene squence of iASPP was consistent with that reported (gi 60457962) in GenBank .The mRNA expression levels of iASPP gene of SW480 and Lovo cell lines which transfect the positive plasmid were increased ,and the cell apoptosis rates were de-creased .Conclusion We successfully constructed the recombinant expression plasmid pcDNA 3 .1(+ )-iASPP ,and the plasmid were successfully expressed in colon carcinoma cell lines SW 480 and Lovo ,the cell apoptosis rates of those cell lines were decreased .These facts indicated that reducing the high expression of iASPP may be a new strategy to renew the abilities of P 53 tumor suppressor .

Objective To explore the clinical effect of applying standard process management in early enteral nutrition support for elderly patients with gastric cancer after operation. Methods A total of142 elderly patients with gastric cancer in our hospital were randomly divided into control group ( n=71) treated with the traditional nursing of enteral nutrition and intervention group ( n=71) adopting the standardized process man-agement, and then enteral nutrition tolerance, complications, nutritional status and recovery of gastrointestinal function were compared between the two groups. Results There was no significant difference in age, sex and preoperative nutritional condition and operation situation between the two groups ( P>0. 05) . The incidence of nutritional intolerance and the cost of nutritional support were lower than those in the control group ( P<0. 05) . The completion rate of enteral nutrition target in the intervention group was significantly higher than that in the control group ( P<0. 05) . The anal exsufflation time and defecation time of patients in the intervention group were shorter than those in the control group ( P<0. 05) . There was no significant difference in pulmonary infec-tion, incision infection and postoperative hospital stay between the two groups ( P>0. 05 ) . Conclusions Standardized process management of early enteral nutrition after operation could significantly improve the toler-ance of enteral nutrition, meanwhile, it has positive significance in promoting rapid recovery of elderly patients with gastric cancer.

Objective: To explore the effect of early enteral nutrition(EEN) and parenteral nutrition(PN) on the postoperative outcomes of patients with gastric cancer and nutritional risk in enhanced recovery after surgery. Methods: A total of 130 patients with gastric cancer hospitalized in department of surgery of Ningbo First Hospitalfrom September 2016 to May 2018 were selected and divided into early enteral nutrition support group (EEN) which was placed with jejunal nutrition tube during the operation, and enteral nutrition started within 12-24 hours after the operation, and parenteral nutrition support group (PN) which was given parenteral nutrition support one day after surgery. Patients in both groups were given nutrients of equal heat and nitrogen.The incidence of nutrition-related complications, the incidence of infection-related complications, the length of postoperative hospital stay and the time of anal exhaust were compared between the two groups. Results: The incidence of nutrition-related complications was 10 cases (15.38%) and 4 cases (6.15%)in EEN group and PN group, that was not statistically different (P=0.157). The incidence of infection-related complications was 3 cases (4.61%) and 5 cases (7.69%) in EEN group and PN group, that was not statistically different (P=0.715). The postoperative hospital stay was 11 days (range, 10-15) and 12 days (range, 11-13)in EEN group and PN group, that was not statistically different (P=0.233). The first anal exhaust time and defecation timewere 64 hours (range, 52-77) and 87 hours (range, 76-100) in EEN group and 72 hours(range, 60-86) and 96 hours(range, 86-120) in PN group, that was statistically different(P=0.001, P=0.034). Conclusion Enhanced recovery after surgery, early enteral nutrition after gastric cancer surgery may promote the recovery of intestinal function, but the complications and hospital stay after operation are not improved.

Objective:To investigate the effect of sarcopenia on physical performance and nutritional status in elderly patients with gastrointestinal tumors.Methods:a total of 120 eligible in-patients were enrolled from gastrointestinal surgery department of a Grade A tertiary hospital in Ningbo city from September 2021 to January 2022. General clinical data were collected, nutritional risk was screened using Nutritional Risk Screening 2002 (NRS 2002), physical performance was assessed by Short Physical Performance Battery (SPPB), skeletal muscle index at the third lumbar vertebra level (L3 SMI) was calculated using abdominal CT scan, and grip strength/muscle strength, gait speed by 6 Meter Timed Walk Test and calf circumference were measured. Subjects were divided into sarcopenia and non-sarcopenia group according to the diagnostic criteria of the Asian Working Group for Sarcopenia (AWGS). Impacting factors of sarcopenia and the correlation between muscle mass and physical performance in elderly patients with gastrointestinal tumors were analyzed.Results:The overall prevalence rate of sarcopenia in elderly patients with gastrointestinal tumors was 28.3% (34/120) and it was 28.9% (22/76) and 27.3% (12/44) in males and females respectively. There were statistically significant differences in age, tumor stage, nutritional risk, body mass index, calf circumference, L3 SMI, grip strength/muscle strength, gait speed and SPPB score between patients with and without sarcopenia (P<0.05). Muscle strength/grip strength, L3 SMI and gait speed were closely correlated with the occurrence of sarcopenia in elderly patients with gastrointestinal tumors (P<0.05). The L3 SMI was positively correlated with physical performance as assessed with SPPB in both groups.Conclusions:Sarcopenia is a common complication in elderly patients with gastrointestinal tumors with multiple influencing factors. Timely nutritional intervention and exercise intervention should be incorporated into the treatment of elderly gastrointestinal tumor patients with sarcopenia in order to improve nutrition status.

Objective:To investigate the influencing factors for microvascular invasion (MVI) in hepatocellular carcinoma based on three-dimensional visualization and the construction of its nomogram model.Methods:The retrospective cohort study method was conducted. The clinico-pathological data of 190 patients with hepatocellular carcinoma who were admitted to Henan University People′s Hospital from May 2018 to May 2021 were collected. There were 148 males and 42 females, aged (58±12)years. The 190 patients were randomly divided into the training set of 133 cases and the validation set of 57 cases by the method of random number table in the ratio of 7:3. The abdominal three-dimensional visualization system was used to characterize the tumor morphology and other imaging features. Observation indicators: (1) analysis of influencing factors for MVI in hepatocellular carcinoma; (2) construction and evaluation of nomogram model of MVI in hepatocellular carcinoma. Measurement data with normal distribution were expressed as Mean± SD, and independent sample t test was used for comparison between groups. Measurement data with skewed distribution were expressed as M( Q1, Q3), and non-parametric rank sum test was used for comparison between groups. Count data were expressed as absolute numbers, and the chi-square test was used for comparison between groups. Corresponding statistical methods were used for univariate analysis. Binary Logistic regression model was used for multivariate analysis. Receiver operator characteristic (ROC) curves were plotted, and the nomogram model was assessed by area under the curve (AUC), calibration curve, and decision curve. Results:(1) Analysis of influencing factors for MVI in hepatocellular carcinoma. Among 190 patients with hepatocellular carcinoma, there were 97 cases of positive MVI (including 63 cases in the training set and 34 cases in the validation set) and 93 cases of negative MVI (including 70 cases in the training set and 23 cases in the validation set). Results of multivariate analysis showed that alpha-fetoprotein, vascular endothelial growth factor, tumor volume, the number of tumors, and tumor morphology were independent factors affecting the MVI of patients with hepatocellular carcinoma ( odds ratio=5.06, 3.62, 1.00, 2.02, 2.59, 95% confidence interval as 1.61-15.90, 1.28-10.20, 1.00-1.01, 1.02-3.98, 1.03-6.52, P<0.05). (2) Construction and evaluation of nomogram model of MVI in hepatocellular carcinoma. The results of multivariate analysis were incorporated to construct a nomogram prediction model for MVI of hepatocellular carcinoma. ROC curves showed that the AUC of the training set of nomogram model was 0.85 (95% confidence interval as 0.79-0.92), the optimal fractional cutoff based on the Jordon′s index was 0.51, the sensitivity was 0.71, and the specificity was 0.84. The above indicators of validation set were 0.92 (95% confidence interval as 0.85-0.99), 0.50, 0.90, and 0.82, respectively. The higher total score of the training set suggested a higher risk of MVI in hepatocellular carcinoma. The calibration curves of both training and validation sets of nomogram model fitted well with the standard curves and have a high degree of calibration. The decision curve showed a high net gain of nomogram model. Conclusions:Alpha-fetoprotein, vascular endothelial growth factor, tumor volume, the number of tumors, and tumor morphology are independent influencing factors for MVI in patients with hepatocellular carcinoma. A nomogram model constructed based on three-dimensional visualized imaging features can predict MVI in hepatocellular carcinoma.