BACKGROUND:Immunotherapy with autologous immune cel s has been developed as a major adjuvant therapy for malignant tumors, but its mechanism of action has not been elucidated. OBJECTIVE:To investigate the relationship between cytokine-induced kil er cel secretion and apoptosis in human liver cancer stem cel s. METHODS:Human liver cancer stem cel s, HepG2 cel s, were isolated and enriched using serum-free suspension method. The peripheral blood mononuclear cel s from patients with liver cancer were induced byγ-interferon, CD3 monoclonal antibody and recombinant human interleukin-2 to form kil er cel s. Passage 1 liver cancer stem cel s were divided into control group (culture alone) and experimental group (co-culture of cytokines-induced kil er cel s and human liver cancer stem cel s). At 48 hours after culture, apoptosis in human liver cancer stem cel s was detected using flow cytometry, and expression of caspase-3 mRNA and protein was detected using RT-PCR and western blot, respectively. RESULTS AND CONCLUSION:The apoptotic rate in the control group was significantly lower than that in the experimental group (P<0.05). The expressions of caspase-3 at mRNA and protein levels were both higher in the experimental group than the control group (P<0.05). Experimental findings show that cytokines-induced kil er cel s can significantly promote apoptosis in human liver cancer stem cel s, and up-regulate the caspase-3 mRNA and protein expressions dramatical y.

Objective Mouse models of sepsis-induced myocardial injury by intraperitoneal injection of lipopolysaccharide (LPS) was established in order to provide a reliable method for the research of pathogenesis of sepsis-induced myocardial injury. Methods According to the method of random number table, a total of 150 male C57BL/6 mice were divided into five groups: NC group, sham group, and LPS 10, 12, 15 mg/kg groups, with 30 in each group. Septic myocardial injury was induced by intraperitoneal injection LPS in mice; sham group was injected with equal 0.9% saline; while there was no treatment in mice of NC group. Fifteen of the 30 mice in each group were used to observe the general status of mice before and after LPS or saline injection. Twenty-four hours after LPS or saline injection, the left ventricular function was assessed by echocardiography, serum level of cardiac troponin (cTnI) was determined by enzyme linked immunosorbent assays (ELISA), and the cardiac histomorphology and ultrastructure were observed; the other 15 mice were used to monitor the 7-day mortality after LPS or saline injection. Results The mice challenged to LPS displayed symptoms of sepsis, such as depression, ruffled fur, and diarrhea. Compared with NC group, left ventricular ejection fraction (LVEF), left ventricular fraction shortening (LVFS) were significantly decreased at 24 hours after LPS administration in LPS 10, 12, 15 mg/kg groups [LVEF: 0.459±0.044, 0.432±0.034, 0.348±0.064 vs. 0.588±0.019, LVFS: (22.36±2.60)%, (20.78±1.91)%, (16.27±3.31)% vs. (30.55±1.30)%, all P < 0.01], and cTnI levels were significantly increased (ng/L: 270.40±43.50, 281.14±41.79, 298.39±42.05 vs. 192.59±16.90, all P <0.01). Myocardium injury was observed in three LPS groups, myocardial fibrosis, interstitial edema, erythrocyte leakage and infiltrating inflammatory cells were observed under light-microscope; ultrastructural changes disorderly arranged in cardiac muscle fibers, mitochondrial swelling and even partly missing mitochondria cristae were found under transmission electron microscope (TEM), and the higher of the dose, the more sever of the damage. There was no significant difference between sham group and NC group. The 7-day mortality in LPS 10, 12, 15 mg/kg groups were 33.3%, 53.3% and 86.7%, respectively, while no death in the NC group and sham group. Conclusion For establishing the mouse model of sepsis-induced myocardial injury, intraperitoneal injection with 12 mg/kg LPS is a preferable choice in our research.

Objective : To investigate the protective effect of adiponectin APN on myocardial injury caused by sep⁃ sis and its possible mechanism. : Methods Results : Compared with sham group , the expression of Bax , cleaved Caspase3 protein increased and the expression of Bcl⁃2 protein decreased in LPS group. Compared with LPS group , the injury in APN + LPS group was significantly alleviated , showing that the expression of Bax , cleaved caspase3 protein decreased and the expression of Bcl⁃2 protein increased. Meanwhile , compared with sham group , the expression of Cx43 increased in LPS group and decreased in APN + LPS group. Conclusion Adiponectin can attenuate LPS⁃induced cardiac injury in septic mice. The mechanism may be through the inhibition of apoptotic signal pathway and the expression of Cx43.