Objective To establish a stable in vitro model of CD8+T cell exhaustion.Methods CD8+T cells were isolated and purified from the spleens of ovalbumin-specific CD8+T cell receptor(OT-I)transgenic mice and subjected to chronic antigen stimulation to induce exhaustion in vitro.Flow cytometry was employed to evaluate the expressions of exhaustion markers,secretion of effector cytokines,and transcription factor profiles in CD8+T cells.Exhausted and effector(non-exhausted)CD8+T cells were co-cultured with tumor cells before tumor cell viability was measured to assess the cytotoxic potential of CD8+T cells.Additionally,N-acetyl-L-cysteine(N-AC)was used as a positive control during exhaustion induction to validate the model.Results Chronic stimulation resulted in a significant upregulation of inhibitory receptors,including programmed cell death protein 1(PD-1),T cell immunoglobulin and mucin domain-containing protein 3(TIM-3),T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motifdomain(TIGIT),and lymphocyte activation gene 3(LAG-3).Concurrently,the secretion of key effector cytokines such as tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)was markedly reduced.Exhausted CD8+T cells exhibited diminished cytotoxicity against tumor cells compared to effector CD8+T cells.Notably,treatment with N-AC effectively restored the function of exhausted CD8+T cells and enhanced their anti-tumor activity.Conclusion This study has established an effective in vitro model for CD8+T cell exhaustion.The use of N-AC demonstrates its potential to restore functionality in exhausted CD8+T cells,underscoring the reliability and utility of this model for investigating the anti-tumor potential of exhausted T cells.

Objective: To analyze the risk factors of delirium in intensive care unit (ICU) patients, and to investigate the predictive value of C-reactive protein (CRP), procalcitonin (PCT), lactic acid (Lac) and neuron-specific enolase (NSE) in the diagnosis of ICU delirium. Methods: The patients admitted to central ICU and respiratory medicine ICU of Changzhou First People's Hospital from August 2016 to November 2017 were enrolled. The patients were divided into two groups according to whether delirium occurred within 7 days or not, which was evaluated by using the confusion assessment method for ICU (CAM-ICU). The gender, age and blood CRP, PCT, Lac, NSE levels were compared between the two groups. Multivariate Logistic regression model was used to analyze the risk factors of ICU delirium. Receiver operating characteristic curve (ROC) was drawn to assess the predictive value of CRP, PCT, Lac and NSE in the occurrence of ICU delirium. Results: 133 patients were enrolled. Delirium occurred in 67 patients, and did not occurred in 66 patients, with a prevalence rate of 50.4%. There was no significant difference in gender or age between the two groups. Compared with non-delirium group, blood CRP, PCT and Lac levels in delirium group were significantly increased [CRP (mg/L): 110.75±77.31 vs. 51.32±36.51, PCT (μg/L): 3.95 (1.01, 23.90) vs. 0.09 (0.06, 0.36), Lac (mmol/L): 2.40 (1.70, 4.30) vs. 1.20 (0.90, 2.00), all P < 0.01], but no significant difference was found in NSE [μg/L: 12.59 (9.61, 17.69) vs. 13.39 (10.14, 19.05), P > 0.05].② It was shown by multivariate Logistic regression analysis that blood PCT and Lac were risk factors of ICU delirium [PCT: odds ratio (OR) = 1.185, 95% confidence interval (95%CI) = 1.006-1.396, P = 0.042; Lac: OR = 1.398, 95%CI = 1.011-1.934, P = 0.043].③ROC curve analysis showed that blood CRP, PCT and Lac had certain predictive value for ICU delirium, and the area under the ROC curve (AUC) of PCT was the highest (0.840 vs. 0.694 and 0.751). When the cut-off value of PCT ≥ 0.55 μg/L, the sensitivity was 72.7%, the specificity was 86.2%, positive predictive value was 84.48%, and negative predictive value was 75.68%. Blood NSE had no predictive value for ICU delirium (AUC = 0.446, P = 0.290). Conclusions Blood PCT and Lac are the risk factors of ICU delirium. PCT has predictive value for ICU delirium.

OBJECTIVE: To analyze the risk factors of delirium in intensive care unit (ICU) patients, and to investigate the predictive value of C-reactive protein (CRP), procalcitonin (PCT), lactic acid (Lac) and neuron-specific enolase (NSE) in the diagnosis of ICU delirium. METHODS: The patients admitted to central ICU and respiratory medicine ICU of Changzhou First People's Hospital from August 2016 to November 2017 were enrolled. The patients were divided into two groups according to whether delirium occurred within 7 days or not, which was evaluated by using the confusion assessment method for ICU (CAM-ICU). The gender, age and blood CRP, PCT, Lac, NSE levels were compared between the two groups. Multivariate Logistic regression model was used to analyze the risk factors of ICU delirium. Receiver operating characteristic curve (ROC) was drawn to assess the predictive value of CRP, PCT, Lac and NSE in the occurrence of ICU delirium. RESULTS: 133 patients were enrolled. Delirium occurred in 67 patients, and did not occurred in 66 patients, with a prevalence rate of 50.4%. (1) There was no significant difference in gender or age between the two groups. Compared with non-delirium group, blood CRP, PCT and Lac levels in delirium group were significantly increased [CRP (mg/L): 110.75±77.31 vs. 51.32±36.51, PCT (μg/L): 3.95 (1.01, 23.90) vs. 0.09 (0.06, 0.36), Lac (mmol/L): 2.40 (1.70, 4.30) vs. 1.20 (0.90, 2.00), all P < 0.01], but no significant difference was found in NSE [μg/L: 12.59 (9.61, 17.69) vs. 13.39 (10.14, 19.05), P > 0.05]. (2) It was shown by multivariate Logistic regression analysis that blood PCT and Lac were risk factors of ICU delirium [PCT: odds ratio (OR) = 1.185, 95% confidence interval (95%CI) = 1.006-1.396, P = 0.042; Lac: OR = 1.398, 95%CI = 1.011-1.934, P = 0.043]. (3) ROC curve analysis showed that blood CRP, PCT and Lac had certain predictive value for ICU delirium, and the area under the ROC curve (AUC) of PCT was the highest (0.840 vs. 0.694 and 0.751). When the cut-off value of PCT ≥ 0.55 μg/L, the sensitivity was 72.7%, the specificity was 86.2%, positive predictive value was 84.48%, and negative predictive value was 75.68%. Blood NSE had no predictive value for ICU delirium (AUC = 0.446, P = 0.290). CONCLUSIONS Blood PCT and Lac are the risk factors of ICU delirium. PCT has predictive value for ICU delirium.
C-Reactive Protein ; Calcitonin ; Calcitonin Gene-Related Peptide ; Delirium ; Humans ; Intensive Care Units ; Prognosis ; Retrospective Studies ; Sepsis