Objective To investigate the apoptotic effect of the transmembrane form vaccine of human blood group A mimotope on malignant melanoma cell line B16. Methods B16 cells were transfected with different recombinant plasmid through Lipofectamine 2000 and incubated with different concentration of monoclonal anti-A antibody at 2.5 μg/ml, 5 μg/ml,10 μg/ml and 20 μg/ml. Apoptosis rate of cells was determined with Annexin Ⅴ/PI double staining by flow cytometry. Results Apoptosis rate to P/F-M-pIRES group B16 cells was 74.74% when anti-A monoclonal antibody concentration was 10 μg/ml; apoptosis rate of plasmids carrying peptide/Fas fusion gene such as P/F-M-pIRES group and P/F-pIRES group were significantly higher than M-pIRES group and pIRES group. The apoptosis rate was statistically significantly different between different recombinated plasmid groups (F＝669.707,P＜0.01). The apoptosis rate was statistically significantly different between different antibody groups (F＝106.596,P＜0.01). The interaction between recombinated plasmid groups and antibody groups was statistically significant (F＝34.806,P＜0.01). Conclusions The transmembrane form vaccine of human blood group A mimotope could induce B16 cell apoptosis in vitro. This vaccine may be a promising candidate for potential malignant melanoma therapy.

Objective To improve the diagnosis rate and decrease misdiagnosis through analyzing the clinical and experimental features of neuroblastoma (NB) in children.Methods The clinical and experimental features associated with the misdiagnosis factors of 22 NB cases were analyzed retrospectively.Results Of the 22 cases,leg pain in 15 cases (68.18 ％),fever in 19 cases (86.36 ％),anemia in 20 cases (90.90 ％),abdominal pain and bloating in 15 cases (68.18 ％),exophthalmoses and bruises eyes in 4 cases (18.18 ％) and easily perspire in 20 cases (90.90 ％).Abdomen was the most common primary site founding in 16 cases (accounting for 72.72 ％),followed by mediastinum founding in 4 cases (accounting for 18.18 ％),other sites were 2 cases (9.09 ％).Laboratory results showed that anemia was the most common cause in 20 cases (90.90 ％),oligoleukocythemia in 2 cases (9.09 ％) and less in thrombocytopenia about 3 cases (13.63 ％).Elevated values were recorded in 77.77 ％ of patients for vanillylmandelic acid (VMA),86.36 ％ for ferritin (Fer),90.90 ％ for lactate dehydrogenase (LDH),and in 100.00 ％ for neuron specific enolase (NSE).20 cases were found metastasis when diagnosed,which accounted for 90.90 ％.16 cases (72.72 ％) were found tumour cells which determined in bone marrow and 20 cases (90.90 ％) were positive for CD56-FITC/CD81-PE/CD45-Percp by flow cytometry (FCM).12 (54.54 ％) of 22 NB cases were misdiagnosed for rheumatoid diseases (4 cases),dyspepsia (4 cases),leukemia (3 cases) and thrambocytopenia (1 case).In 22 cases,20 cases (90.90 ％) were positive for CT scan.16 cases (72.72 ％) were positive for abdominal B ultrasonography,and 14 cases (63.63 ％) were positive for radionuclide bone scan.Conclusion Bultrasonography and CT scan will contribute to find the primary focus of NB at early stage.VMA,bone marrow smear,biopsy and FCM detection could reduce misdiagnose rate.

Objective To investigate the trend of cytokines in patients with hemophagocytic lymphohistiocytosis (HLH) and analyze its significance.Methods 16 patients with HLH from January 2011 to May 2013 were selected.The patients were divided into remission group and death group by prognosis.Serums of the two groups were collected when they were hospitalized and at 7 th,14 th,21st,28th and 42nd day during chemotherapy,and they were fractionated HLH 1-6 groups and HLH a-d groups again,then the levels of IL-18,IL-10,IL-12,NF-κB,TNF-α and neopterin were tested by enzyme linked immuno sorbent assay (ELISA) to analysis their trend.Results The levels of all of cytokines in the remission group declined with chemotherapy,the difference between HLH1 group and another HLH groups was statistically significant (P ＜ 0.05).In the death group,the levels of NF-κB,IL-12 and neopterin had no downward trend with chemotherapy,and the difference between HLHa group and another HLH groups was not statistically significant (all P ＞ 0.05).The level of TNF-α declined with chemotherapy,and the differences between HLHa group and HLHc group,HLHa group and HLHd group were statistically significant (P =0.049,0.000).The level of IL-10 declined sharply in the first week of chemotherapy,and the difference between HLHa group and HLHb group was statistically significant (P =0.00).The level of IL-18 declined after the 2nd-weeks' chemotherapy,and the differences between HLHa group and HLHb group,HLHa group and HLHc group were statistically significant (P =0.03,0.02).Conclusions In the remission patients,the levels of serum IL-18,IL-10,IL-12,NF-κB,TNF-α and neopterin declined after chemotherapy.In the death patients,the downward trend is not obvious.It was preliminarily confirmed that the prognosis of HLH is related to the trend of cytokines during chemotherapy.

Objective:To analyze the potential role and prognostic value of platelet-to-lymphocyte ratio (PLR) at an early stage in arterial-venous extracorporeal membrane oxygenation (VA -ECMO).Methods:Totally 83 adult patients with VA-ECMO from June 2018 to June 2020 treated at Emergency Department of Jiangsu Provincial Hospital were retrospectively analyzed. Baseline characteristics between survivors ( n=46) and non-survivors ( n=37) were compared. Logistic regression analysis was used to predict the risk factors associated with 28-day mortality in VA-ECMO patients. The cut-off value was calculated by the receiver operating characteristic (ROC) curve. Results:PLR48-h ( OR=1.018,95% CI: 1.001-1.036, P=0.039) and continuous renal replacement therapy (CRRT) ( OR=7.095,95% CI: 1.099-45.799, P=0.039) were relevant risk factors of 28-day mortality in VA-ECMO patients. The cut-off value of PLR48-h was 156.3 [sensitivity: 57.8%, specificity: 86.1%, and area under the curve (AUC): 0.756]. Compared with the high PLR group (>156.3), the incidences of acute kidney injury (AKI) ( P<0.001) and bleeding events ( P=0.013) were significantly higher in the low PLR group (<156.3). Conclusions:The early PLR reduction and CRRT application during VA-ECMO support are related to poor prognosis.

Objective:To investigate the clinical significance of the acute physiology and chronic health evaluationⅡ (APACHEⅡ) combined with different systematic inflammation markers (SIMs) including neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lymphocyte-to-monocyte ratio (LMR)-in adult patients with venous-arterial extracorporeal membrane oxygenation (VA-ECMO).Methods:A total of 89 adult patients with VA-ECMO ( ≥ 3 d) in the Emergency Department of Jiangsu Provincial People's Hospital from January 2017 to June 2020 were retrospectively analyzed. Patients were divided into two groups: survivors ( n=39) and non-survivors ( n=50). The baseline APACHE Ⅱscore and PLR, NLR, LMR before ECMO implantation and at 1, 2, 3 day after ECMO were recorded. Binary logistic regression was used to analyze the risk factors of 28-day mortality in patients with VA-ECMO. The utility of APACHEⅡ score and SIMs alone or combination for predicting clinical prognosis was evaluated using receiver operating characteristic (ROC) curve analysis. The patients were divided into the high risk group and the low risk group according to the best cut-off value, and the difference of ECMO-related complications between the two groups was compared. Results:When combined APACHEⅡ score with SIMs, APACHEⅡ + PLR 48 h + LMR 24 h + LMR 72 h demonstrated the greatest predictive ability with an AUC of 0.833. Compared with the high-risk group, the low-risk group has a lower incidence of acute renal injury, infection, bleeding complications, the use of continuous renal replacement therapy, mechanical ventilation, and a higher hospital survival rate.Conclusions:The combination of APACHEⅡ score and SIMs-PLR, LMR- is better than a single one for death prediction, and it is expected to be a new predictive model for early identification of the risk of death or poor prognosis in patients with VA-ECMO.

Objective:To investigate the safety of early whole body computed tomography (WBCT) combined with coronary angiography (CAG) in patients with extracorporeal cardiopulmonary resuscitation (ECPR) and its application value in the diagnosis of cardiac arrest and complications of cardiopulmonary resuscitation (CPR).Methods:This was a retrospective study. Patients who underwent ECPR in the Emergency Department of the First Affiliated Hospital of Nanjing Medical University from January 2017 to July 2021 were enrolled in this research. Patients younger than 18 years or with incomplete clinical data were excluded. The results of WBCT and CAG examinations after ECPR were collected.Results:A total of 89 patients with ECPR, aged (47±17) years, were enrolled in the study, all underwent WBCT examination, and no adverse events such as ECMO and tracheal tube shedding occurred. WBCT found 7 cases of pulmonary embolism, 3 cases of aortic dissection and 2 cases of cerebral hemorrhage. WBCT identified CPR-related complications in 42 cases, including rib fractures ( n=20), pneumothorax ( n=5), mediastinal emphysema ( n=5), subcutaneous emphysema ( n=6), and hematoma or swelling at puncture site ( n=6). Fifty-five patients underwent CAG examination, the most common culprit vessels were the left anterior descending branch disease (58.2%) followed by the left circumflex branch disease (27.3%), the right coronary artery disease (21.8%) and left main artery disease (12.7%). Conclusions:Early WBCT and CAG examinations are of great significance and safety for the guidance of treatment in ECPR patients.

Objective:To explore the prognostic value of survival after veno-arterial ECMO (SAVE) score combined with 24-h lactate on the machine in patients with extracorporeal cardiopulmonary resuscitation (ECPR).Methods:Totally 59 patients treated with ECPR in the Emergency Department of the First Affiliated Hospital of Nanjing Medical University from April 2017 to June 2021 were retrospectively analyzed. According to the 28-day prognosis, the patients were divided into the death group ( n=36) and the survival group ( n=23). The differences in baseline data were analyzed, and multivariate logistic regression was performed to identify the influencing factors of 28-day mortality in patients with ECPR. The receiver operating characteristic (ROC) curve was applied to evaluate the predictive value of SAVE score, 24-h lactate and their combined detection for predicting 28-day mortality risk in patients with ECPR. Results:The 28-day survival rate of patients with ECPR was 39% (23/59). SAVE score of the death group was significantly lower than that of the survival group (-11.67±4.60 vs. -2.43±4.77, P<0.001), and the 24-h lactate in the death group was significantly higher than that in the survival group [5.94 (3.37, 12.40) mmol/L vs. 1.65 (1.07, 3.15) mmol/L, P<0.001]. Multivariate logistic regression analysis showed that SAVE score ( OR=0.703, 95% CI: 0.566-0.873, P=0.001) and 24-h lactate ( OR=1.608, 95% CI: 1.025-2.523, P=0.039) were independent influencing factors of 28-day mortality in ECPR patients. ROC curve analysis showed that the best cut-off value of SAVE score was -6, the sensitivity was 78.30% and specificity was 91.70%. The best cutoff value of 24-h lactate was 4.7 mmol/L, the sensitivity was 63.90% and specificity was 100.00%. The sensitivity and specificity of the combined detection of SAVE score and 24-h lactate were 82.60% and 100.00%, respectively. The area under the curve (AUC) of SAVE score combined with 24-h lactate for predicting the 28-day mortality risk in patients with ECPR was larger than that of SAVE score and 24-h lactate alone (0.952 vs. 0.917; 0.952 vs. 0.847). Conclusions:Lower SAVE score and higher 24-h lactate are independently risk factors of 28-day mortality in patients with ECPR, and SAVE score combined with 24-h lactate on the machine has a good predictive value for the prognosis of patients with ECPR.