This paper describes the elaboration of the use of HRP labelled Gangliosides as second antibody to the tentative identification of Cholera enterotoxin(toxoid)and Tetanus toxin(toxoid).The method is called Enzyme linked Ganglioside Immunesorbent assay (ELGISA).The sensitivity of this was campared with those of Ouchlerlony double diffusion method,VCS and TLA,but found to be more sensitive.

AIM:To investigate the effects of celecoxib on viability , apoptosis and autophagy in acute myeloid leukemia (AML) cell lines HL-60 and HL-60A.METHODS:The HL-60 cells and HL-60A cells were cultured with vari-ous concentrations (0, 20, 40, 60, 80 and 100μmol/L) of celecoxib.The inhibitory effect of celecoxib on the cell viabil-ity was evaluated by MTT assay .Apoptosis was analyzed by Annexin-V/PI staining.Apoptosis-related and autophagy-relat-ed proteins were determined by Western blot .RESULTS:IC50 of celecoxib were 49.4 μmol/L, 32.0 μmol/L and 25.1μmol/L for HL-60 cells treated with celecoxib for 24 h, 48 h and 72 h, respectively.For HL-60A cells, the corresponding IC50 were 69.1 μmol/L, 42.5 μmol/L and 29.6 μmol/L, respectively.The results of flow cytometry analysis showed the proportions of Annexin-Ⅴ+PI-, Annexin-Ⅴ+PI+and Annexin-Ⅴ-PI+cells were increased in the HL-60 cells, and those of Annexin-Ⅴ+PI-and Annexin-Ⅴ+PI+cells were increased in the HL-60A cells treated with celecoxib for 24 h. After treated with celecoxib , the induction of apoptosis was observed , the apoptosis-related proteins cleaved caspase-3 and cleaved PARP were upregulated , the autophagy-related proteins LC3 II and P62 were both increased , and mTOR, p-mTOR, 4-EBP and p-4-EBP were not changed , indicating that celecoxib inhibited autophagy in the AML cells without the mTOR pathway involvement .CONCLUSION:Celecoxib inhibits the viability of HL-60 cells and HL-60A cells in a time-and dose-dependent manner by its effects of inducing apoptosis and necrosis .Celecoxib inhibits mTOR-independent autoph-agy in AML cells, indicating a possible way of using celecoxib for enhancing the antitumor activity of therapeutic agents to induce cytoprotective autophagy in the AML cells .

Objective: To explore the status of lymph node metastasis of middle third thoracic esophageal squamous cell carcinoma and its influence on the prognosis and to seek the reasonable range of lymphade-nectomy. Methods: A total of 129 patients who underwent curative esophagectomy with modern two-field lymphadenectomy of middle third thoracic esophageal squamous cell carcinoma were reviewed. Results: The lymph node metastasis rate was 56.6% and the upper mediastinal lymph node metastasis rate was 43.4%. The lymph node metastasis ratio (positive nodes/total dissected nodes, LMR) was 11.3%. Paraesophageal lymph nodes, lymph nodes near the right recurrent nerve, the left gastric and infracadnal lymph nodes were most commonly involved when the tumor was located in the middle thoracic esophagus. Tumor differentiation, the depth of tumor invasion and the length of tumor were influencing factors for lymph node metastasis. The 5-year survival of N_0, N_1 (LMR≤20%) and N_1 (LMR>20%) patients were 50.4%, 31.0% and 6.8%, respective-ly, with a significant difference among the three groups (P=0.000). Conclusion: LMR was one of the key fac-tors affecting the prognosis, of esophageal cancer. Patients with middle third thoracic esophageal carcinoma should be treated with radical surgery with modern two-field lymphadenectomy.

Objective To investigate the predictive value of preoperative albumin-to-fibrinogen ratio (A/F) for postoperative survival of esophageal squamous cell carcinoma after radical esophagectomy.Methods The retrospective cohort study was conducted.The clinicopathological data of 559 patients with esophageal squainous cell carcinoma who underwent radical resection in the Sun Yat-sen University Cancer Center from January 2009 and January 2012 were collected.There were 443 males and 116 females,aged from 53 to 66 years,with an average age of 59 years.Of 559 patients,394 with A/F ＞ 11.14 were in high A/F group,and 165 with A/F ≤ 11.14 were in low A/F group.Levels of plasma albumin and fibrinogen were measured at two weeks before surgery,and patients underwent radical esophagectomy.Observation indicators:(1) clinicopathological characteristics of patients;(2) survival of patients;(3) risk factors analysis of prognosis of patients.Follow-up was conducted by outpatient examination and telephone interview to detect postoperative survival of patients up to November 2018.The overall survival time was from surgery date to date of endpoint events including death or the last follow-up,and the overall disease-free survival time was from surgery date to date of endpoint events including tumor recurrence,tumor-falted death,or the last follow-up.Measurement data with skewed distribution were expressed by M (range).Count data were described as percentage,and comparison between groups was analyzed using chi-square test or Fisher exact probability.Comparison of ordinal data was analyzed by Mann-Whitney U nonparametric test.The survival rate and curve were calculated and drawn by Kaplan-Meier method and the Log-rank test was used for survival analysis.The univariate and multivariate analyses were done using COX proportional hazard model.Results (1) Clinicopathological characteristics of patients:of 559 patients,cases with age ≤ 60 years and ＞ 60 years,cases in T1 stage,T2 stage,T3 stage(depth of tumor invasion)were 246,148,60,79,255 in the high A/F group,and 79,86,5,32,128 in the low A/F group,there were statistically significant differences in the age and depth of tumor invasion between the two groups (x2 =10.127,Z=-3.468,P＜0.05).(2) Survival of patients:559 patients were followed up for 97 months (range,91-103 months).The 5-year overall survival rate and 5-year disease-free survival rate were 55.8％ and 48.7％ in the high A/F group,and 38.8％ and 35.8％ in the low A/F group,respectively,with statistically significant differences between the two groups (x2 =16.501,11.679,P＜0.05).(3) Risk factors analysis of prognosis of patients:results of univariate analysis showed that sex,age,preoperative fibrinogen level,preoperative A/F level,surgical method,depth of tumor invasion,tumor pathological N staging,and postoperative adjuvant therapy were associated with 5-year overall survival rate of esophageal squamous cell carcinoma after radical esophagectomy [hazard ratio (HR) =1.362,1.358,1.421,0.617,0.772,1.490,1.732,1.436,95％ confidence interval (CI):1.010-1.835,1.084-1.700,1.114-1.814,0.487-0.781,0.612-0.973,1.239-1.792,1.552-1.934,1.128-1.829,P＜ 0.05];age,preoperative fibrinogen level,preoperative A/F level,surgical method,depth of tumor invasion,tumor pathological N staging,postoperative adjuvant therapy were associated with 5-year disease-free survival rate of esophageal squamous cell carcinoma after radical esophagectomy (HR=1.248,1.371,0.675,0.740,1.427,1.665,1.606,95％CI:1.006-1.547,1.086-1.732,0.538-0.847,0.592-0.924,1.202-1.695,1.498-1.851,1.275-2.022,P＜ 0.05).Results of multivariate analysis showed that age,preoperative A/F level,surgical method,depth of tumor invasion,and tumor pathological N staging were independent factors for 5-year overall survival rate and 5-year disease-free survival rate of esophageal squamous cell carcinoma after radical esophagectomy (HR=1.491,0.699,0.741,1.353,1.761,95％CI:1.184-1.877,0.550-0.888,0.587-0.935,1.120-1.634,1.573-1.971,P＜0.05;HR=1.372,0.760,0.703,1.281,1.692,95％CI:1.100-1.711,0.603-0.957,0.562-0.880,1.074-1.527,1.518-1.887,P＜0.05).Conclusion Preoperative A/F level has a good predictive value for survival of esophageal squamous cell carcinoma after radical esophagectomy,and preoperative A/F level ≤ 11.14 is a independent risk factor for 5-year overall survival rate and 5-year disease-free survival rate of esophageal squamous cell carcinoma after radical esophagectomy.

Glutamate-ammonia ligase (GLUL, also known as glutamine synthetase) is a crucial enzyme that catalyzes ammonium and glutamate into glutamine in the ATP-dependent condensation. Although GLUL plays a critical role in multiple cancers, the expression and function of GLUL in gastric cancer remain unclear. In the present study, we have found that the expression level of GLUL was significantly lower in gastric cancer tissues compared with adjacent normal tissues, and correlated with N stage and TNM stage, and low GLUL expression predicted poor survival for gastric cancer patients. Knockdown of GLUL promoted the growth, migration, invasion and metastasis of gastric cancer cells in vitro and in vivo, and vice versa, which was independent of its enzyme activity. Mechanistically, GLUL competed with β-Catenin to bind to N-Cadherin, increased the stability of N-Cadherin and decreased the stability of β-Catenin by alerting their ubiquitination. Furthermore, there were lower N-Cadherin and higher β-Catenin expression levels in gastric cancer tissues compared with adjacent normal tissues. GLUL protein expression was correlated with that of N-Cadherin, and could be the independent prognostic factor in gastric cancer. Our findings reveal that GLUL stabilizes N-Cadherin by antagonizing β-Catenin to inhibit the progress of gastric cancer.