In order to study T-bet function in mouse cells, a novel retroviral vector expressing mouse T-bet and reporter gene Thy1.1 was constructed. Retrovirus particles were then produced by transfection of the recombinant retroviral plasmid into a packaging cell line Platinum-E. The recombinant retrovirus played considerable infection ability. T-bet expression was then identified by FACS after infection of CD4+ primary T cells from T-bet knockout mouse with recombinant retrovirus. To determine if exogenous expressing T-bet has normal function, we checked the expression level of T-bet target gene, Ifng. IFN-y expression was upregulated in the T-bet knockout T cells infected with recombinant retrovirus. In conclusion, we successfully constructed an effective mouse T-bet recombinant retroviral vector.
Animals ; Cell Line ; Genetic Vectors ; Interferon-gamma ; metabolism ; Mice ; Mice, Knockout ; Recombinant Proteins ; biosynthesis ; Retroviridae ; T-Box Domain Proteins ; biosynthesis ; T-Lymphocytes ; metabolism ; Transfection

Objective To investigate the characteristics of growth and angiogenesis of colon cancer xenograft in nude mice with metabolic syndrome induced by high fat diet.Methods Female BALB/C nude mice were fed with high fat diet (45.0％ from fat,HFD group) or common diet (13.8％ from fat,CD group) for 12 weeks (n=15,respectively).Colon cancer cell line SW480 was marked with green fluorescent protein (GFP) and subcutaneous xenograft model was established.The tumor growth was observed by the in vivo imaging system in small animal at the 4th week.By the end of the experiment,serum glucose and lipid level of the two groups were measured,visceral subcutaneous and visceral adipose tissue,liver and xenograft tumor were dissociated and weighted.The differences of proliferating cell nuclear antigen (PCNA) and CD31 expression in the tumors between groups were analyzed.The t-test or x2 test were performed for group comparison.Results Compared with CD group,the body weight,blood serum glucose level,triglyceride and cholesterol level,adipose content of subcutaneous and visceral of the HFD group significantly increased (t=2.91,4.12,4.43,3.92,3.77 and 4.02,all P＜0.05).Averagedaily energy intake of HFD group was significantly higher than that of CD group (t=2.34,P＜0.05).There was no significant difference between the two groups in liver weight (t=1.02,P＞0.05).However,by HE staining lipid vacuoles in the liver tissue was obvious in HFD group.Average bioluminescent index,tumor volume and weight of xenografts of HFD group were remarkably higher than those of CD group (t=8.84,2.48 and 2.86,all P＜0.05).The immunohistochemical staining results indicated that the strong positive rate of PCNA in xenografts of HFD group was 80.00％ and the microvessel density (MVD) was (25.75±0.96)/per high power field,both of which were higher than those of CD group (14.29％ and (13.33±1.53)/per high power field respectively,x2 =12.52,t=13.35,both P＜0.01).Conclusions The colon cancer xenograft in nude mice with metabolic syndrome induced by high fat diet had a high MVD and grew fast.

Objective:To investigate the predictive value of serum vascular endothelial growth factor (VEGF), squamous cell carcinoma-associated antigen (SCCAg) and miRNA let-7a in lymph node metastasis and postoperative recurrence in patients with laryngeal cancer.Methods:A total of 82 patients with laryngeal cancer in the Second Central Hospital of Baoding from November 2017 to October 2019 were selected as the research subjects, including 18 cases of lymph node metastasis (metastasis group) and 64 cases of non metastasis (non metastasis group). The blood routine was tested before operation, and the baseline data, serum VEGF, SCCAg and miRNA let-7a levels were compared between the two groups. Logistic regression was used to analyze the related influencing factors of lymph node metastasis in patients with laryngeal cancer. The correlation between serum VEGF, SCCAg, miRNA let-7a levels and clinicopathological characteristics was analyzed. The receiver operating characteristic (ROC)curve was used to analyze the value of each index and the combined diagnosis of lymph node metastasis in patients with laryngeal cancer. After 1 year of follow-up, the serum VEGF, SCCAg and miRNA let-7a levels of patients with or without recurrent laryngeal cancer were compared. ROC curve was used to evaluate the value of VEGF, SCCAg, and miRNA let-7a in predicting the recurrence of laryngeal cancer.Results:There were statistically significant differences in tumor node metastasis (TNM) stage, degree of infiltration, degree of differentiation, serum VEGF, SCCAg, and miRNA let-7a levels between the metastasis group and non metastasis group (all P<0.05). Serum VEGF, SCCAg, miRNA let-7a levels in patients with laryngeal cancer were related to TNM stage, degree of infiltration and degree of differentiation (all P<0.05). The combined diagnosis of serum VEGF, SCCAg and miRNA let-7a levels in the diagnosis of lymph node metastasis in patients with laryngeal cancer showed that the diagnostic sensitivity and specificity were 88.89% and 70.31%, respectively. The serum VEGF and SCCAg levels of patients with recurrence after operation were higher than those without recurrence, and the level of miRNA let-7a was lower than those without recurrence (all P<0.05). The sensitivity and specificity of combined serum VEGF, SCCAg and miRNA LET-7a levels in predicting postoperative recurrence of laryngeal cancer were 72.97% and 91.11%, respectively. Conclusions:VEGF, SCCAg, miRNA let-7a in patients with laryngeal cancer have a certain correlation with clinicopathological characteristics, which can assist in the diagnosis of lymph node metastasis and help clinical prediction of postoperative recurrence in patients with laryngeal cancer, and provide a reference for the formulation of clinical treatment plans.

Objective: To investigate the prognostic factors of hyperamylasemia following pancreaticoduodenectomy (PD) . Methods: Clinical data of 359 patients were collected prospectively who underwent PD by the same group at Changhai Hospital of Navy Medical University from January 2017 to June 2018.There were 212 males and 147 females.The median age was 63 years old (range: 23 to 82 years old) .According to whether the patient′s serum amylase was greater than 120 U/L at 0 or 1 day after surgery,the patients were divided into hyperamylasemia group and non-hyperamylasemia group. Univariate analysis and multivariate analysis were used to find out the prognostic factors of hyperamylasemia after PD. Results: Of the 359 patients, 238 cases (66.3%) developed hyperamylasemia.The incidence rate of clinically related pancreatic fistula (15.1% vs.2.5%, P<0.01) , grade B/C post pancreatectomy hemorrhage (8.8% vs. 2.5%, P<0.01) , and surgical site infection (9.2% vs. 3.3%, P=0.04) was significantly higher in the hyperamylasemia group.The severity of complications (CD grade≥Ⅲ: 11.3% vs.4.1%, P=0.023) and postoperative hospital stay (11 days vs. 9 days, P=0.001) were higher in the hyperamylasemia group.In the multivariate analysis, the main pancreatic duct diameter (MPD) ≤3 mm (OR=4.469, 95% CI: 2.563-7.793, P<0.01) , pathological type of disease (pancreatic cancer or pancreatitis) (OR=0.230, 95% CI: 0.122-0.436, P<0.01) and soft texture of pancreas (OR=3.297, 95%CI: 1.930-5.635, P<0.01) were independent prognostic factors for hyperamylasemia. Conclusions Post-PD hyperamylasemia increased the incidence and severity of postoperative complications after PD.MPD≤3 mm, soft texture of pancreas and pathological type of disease were independent prognostic factors of hyperamylasemia.