Objective To explore the effects and mechanism of bexarotene in combination with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) on apoptosis of leukemic cell line KG1a. Methods KG1a cells at logarithmic growth phase were obtained, and were divided into TRAIL group, bexarotene group, 300 ng/mL TRAIL in combination with bexarotene group and 2.0 μmol/L bexaroten in combination with TRAIL group. Cell apoptosis rate was detected in each group by flow cytometry. Flow cytometry was also employed to determine the apoptosis rates of KG1a cells after treatment with bexarotene and TRAIL in different sequences. The expression of Fas associated death domain-like IL-1 beta converting enzyme inhibitory protein (c-FLIP) was detected by Western blotting. Results There was no significant difference in cell apoptosis rates between TRAIL group and bexarotene group of each concentration (except for bexarotene 2.0 μmol/L) (P > 0.05). The cell apoptosis rates of 300 ng/mL TRAIL in combination with bexarotene group and 2.0 μmol/L bexaroten in combination with TRAIL group were significantly higher than those in TRAIL group and bexarotene group of each corresponding concentration (P <0.01). Sequential analysis revealed that bexarotene could reverse the resistance of KG1a cells to TRAIL (P < 0.001). Compared with single use of 2.0 μmol/L bexarotene or 300 ng/mL TRAIL, combination use could significantly down-regulated the expression of c-FLIP (P < 0.05). Conclusion Bexarotene can significantly enhance the apoptosis of KG1a cells induced by TRAIL, which may be attributed to the down-regulation of c-FLIP expression.

Objective To evaluate the sufentanil-sparing effect of ketorolac tromethamine for postoperative analgesia in the elderly patients.Methods Sixty ASA Ⅱ or Ⅲ patients,aged ≥ 65 yr,with a body mass index of 18-24 kg/m2,undergoing elective gynecological operations,were randomly divided into 2 groups(n =30 each):sufentanil group(group S)and ketorolac tromethamine plus sufentanil group(group T).Both groups received combined intravenous-inhalational anesthesia and patient-controlled intravenous analgesia(PCIA)after operation.PCIA solution contained ketorolac tromethamine 180 mg and sufentanil 100 μg in 100 ml of normal saline in group T.After a loading dose of ketorolac tromethamine 30 mg was injected intravenously at 15 min before the end of operation,the PCA pump was set up with a 1.6 ml bolus dose,a 20 rain lockout interval and background infusion at a rate of 1.5 ml/h in group T.PCIA solution contained sufentanil 100 μg in 100 ml of normal saline in group S.After a loading dose of sufentanil 5 μg was injected intravenously at 15 min before the end of operation,the PCA pump was set up with a 1.6 ml bolus dose,a 20 min lockout interval and background infusion at a rate of 1.5 ml/h in group S.The effective analgesia(postoperative VAS scores at rest and during activity ＜ 3)was maintained within 48 h after operation.The amount of sufentanil consumed within 48 h after operation and adverse effects were recorded.Results Compared with group S,the amount of sufentanil consumed within 48 h after operation was significantly reduced,and the incidence of nausea and vomiting,urinary retention and pruritus was significantly decreased in group T(P ＜ 0.05).Conclusion Ketorolac tromethamine used with PCIA with sufenlanil has a significant sufentanil-sparing effecl for posloperative analgesia and improves the safety of analgesia in the elderly patients.

Objective To explore the feasibility and efficacy of the selective portal vein embolization (SPVE) before radiofrequency ablation(RFA) for liver tumor large than 3 cm.Methods 63 patients with 63 liver tumor (＞3 cm) located in single liver segment completely or mostly underwent RFA.21 patients (21 lesions) were randomly assigned to receive SPVE before ablation (SPVE + RFA group),other 42 patients were treated with RFA only (RFA group).The complications and treat results of two groups were collected and compared.Results SPVE were achieved in 20 of 21 patients,and no critical complication were happened in both group.During a observation period of median 14.2 months,local tumor progression were observed in 17 of 42 patients (40.5％) in RFA group and in 3 of 20 patients (15.0％) in SPVE+ RFA group,there were significant difference between two groups(P =0.043).Conclusions SPVE can safely and effectively improve the efficacy of RFA for the liver tumors which large than 3 cm and located in single liver segment.

Objective:To investigate the role and significance of NUFIP-1-mediated ribophagy in apoptosis of dendritic cells (DCs) stimulated by lipopolysaccharide (LPS).Methods:Cultured mouse dendritic cell line DC2.4 were divided into the blank control group and LPS stimulation groups for 6, 12, 24, 48 and 72 h ( n=5). LPS subgroups were consistently cultured with 1 μg/mL LPS for the corresponding incubation time. Western blot was adopted to detect the expression levels of NUFIP-1 and autophagy-related proteins p62 and LC3B across groups. Laser scanning confocal microscopy (LSCM) was applied to detect the expression and cellular localization of NUFIP-1, with its co-localization with Lyso-tracker and LC3B, respectively. The silencing blank vector NS and silencing virus vector NUFIP-1 siRNA were transferred into DC2.4 ( n=3) and stimulated with 1 μg/mL LPS for 24 h. The apoptosis of DC2.4 was measured by flow cytometry analysis. The expression levels of apoptosis-related proteins were determined using Western blot, including cleaved caspase-3 and Bcl-2. One-way analysis of variance (ANOVA) was applied for comparison among multiple groups, and LSD-t method was used for subsequent pairwise comparison. A P<0.05 was considered statistically significant. Results:The results of Western blot showed that expression level of NUFIP-1 in DC2.4 revealed a trend of first increasing and subsequent decreasing upon LPS stimulation for different times (6, 12, 24, 48 and 72 h), and the expression level of NUFIP-1 in the LPS 24 h group was significantly higher than that in the blank control group [blank control group: (0.6786 ± 0.0820); LPS 24 h group: (1.4830 ± 0.1170); P<0.01]. Meanwhile, p62 expression in the LPS 24 h group was significantly lower than that in the blank control group [blank control group: (0.9087 ± 0.1235); LPS 24 h group: (0.3113 ± 0.5571); P<0.01]. Moreover, the conversion from LC3B-I to LC3B-II in the LPS 24 h group was significantly higher than that in the blank control group [blank control group: (0.5542 ± 0.1248); LPS 24 h group: (2.5310 ± 0.3119); P<0.01]. LSCM indicated that NUFIP-1 was predominantly located in the nucleus and perinuclear area in DC2.4. The fluorescence intensity of NUFIP-1 increased in a time-dependent manner from 6 h to 24 h after LPS stimulation, whereas a significant reduction could be observed at 48 h and 72 h after LPS stimulation. Meanwhile, the co-localization of NUFIP-1 with Lyso-tracker and LC3B was substantially reinforced in comparison with the blank control group. Transfection of NUFIP-1 siRNA through lentivirus transfection technology significantly down-regulated the expression level of NUFIP-1 in DC2.4, with statistical differences compared with the blank control group and empty vector group [blank control group: (0.6627 ± 0.1707); empty vector group: (0.6966 ± 0.1107); siRNA group: (0.1428 ± 0.0296); P<0.05]. Flow cytometry analysis revealed that the apoptotic rate of LPS-stimulated DC2.4 was significantly higher in the NUFIP-1 siRNA transfection group than that in the blank control group and empty vector group [blank control LPS 24 h group: (47.91% ± 1.006%); empty vector LPS 24 h group: (70.26% ± 1.011%); siRNA LPS 24 h group: (80.23% ± 2.094); P<0.01]. Western blot analysis of apoptosis-related protein further confirmed that the expression level of cleaved caspase-3 was significantly elevated in the NUFIP-1 siRNA transfection group compared to those of the blank control group and empty vector group under LPS challenge [blank control LPS 24 h group: (0.4748 ± 0.0876); empty vector LPS 24 h group: (0.2849 ± 0.0418); siRNA LPS 24 h group: (0.9733 ± 0.0525); P<0.01]. Likewise, expression of Bcl-2, an anti-apoptotic protein was significantly down-regulated in the siRNA LPS 24 h group [blank control LPS 24 h group: (0.7810 ± 0.0490); empty vector LPS 24 h group: (0.8292 ± 0.0729); siRNA LPS 24 h group: (0.3957 ± 0.0838); P<0.05]. Conclusions:NUFIP-1-mediated ribophagy is significantly activated in DC2.4 upon LPS stimulation, exerting an underlying protective effect on apoptosis.

Objective To summarize the clinical characteristics of adult patients with residual yolk duct,and to explore the diagnosis and treatment strategy of residual vitelline duct in adults.Methods A retrospective analysis on 11 adult cases with residual vitelline duct in our hospital between June 2012 and May 2017 was carried out.Results 8 cases were males,3 cases were females,and median age was 50 years (18-57 y).2 cases were vitelline cyst,9 cases were Meckel diverticulum.2 cases were with ectopic tissue,3 cases with ulcer bleeding,1 case with secondary intra-abdominal hernia and intestinal obstruction,2 cases with secondary infection.The pathological diagnosis of Meckel diverticulum was consistent with preoperative diagnosis.There were no major postoperative complications.The patients were followed up from 6 months to 2 years.Conclusion Most of the residual vitelline duct in adults are Meckel diverticulum and vitelline duct cyst.Resection of residual vitelline duct is the main treatment method.

Objective:To investigate the effect of stress-induced protein Sestrin2 (SESN2) on necroptosis of mouse dendritic cell (DC) induced by lipopolysaccharide (LPS) combined with zVAD, a panaspartate-specific cysteine protease (caspase) inhibitor.Methods:The DC2.4 cell line derived from the bone marrow of mouse in the 3rd to 10th generations was cultured. The cells were stimulated with LPS for 0 hour, 6 hours, 12 hours, and 24 hours, and grouped according to the stimulation time points. Western blotting was performed to determine the protein expression of SESN2 in each group. Overexpression empty lentivirus (NC), SESN2 gene overexpression RNA sequence lentivirus (SESN2 LV-RNA), small interfering empty lentivirus (NS), and SESN2 gene small interfering RNA sequence lentivirus (SESN2 siRNA) were transfected into DC2.4 cells. After 72 hours of transfection, cell fluorescence expression was observed under the inverted fluorescence microscope. Cells in each transfection group were stimulated with LPS for 24 hours. The blank control groups were set up and cultured with phosphate buffered saline (PBS) for 24 hours. Western blotting was performed to measure SESN2 protein expression. In the same groups as above, cells were stimulated with LPS+zVAD for 24 hours. The blank control groups were set up and cultured with PBS for 24 hours. Western blotting was used to determine the expression of mixed lineage kinase domain-like protein (MLKL) and phosphorylated-MLKL (p-MLKL). The p-MLKL levels and the number of positive cells were observed using laser scanning confocal microscopy. The necroptotic cell ratios were assessed by both flow cytometry and Hoechst staining.Results:Compared to the LPS 0 hour group, the expression of SESN2 in the LPS 24 hours group showed a significant increase. Therefore, 24 hours was chosen as the subsequent stimulation time point. After successful lentivirus transduction and 24 hours of cultivation, the MLKL phosphorylation level in the SESN2 siRNA+LPS+zVAD group was significantly higher than that in the NS+LPS+zVAD group. The MLKL phosphorylation in the SESN2 LV-RNA+LPS+zVAD group was significantly lower than that in the NC+LPS+zVAD group. The MLKL phosphorylation levels in both the NS+LPS+zVAD group and the NC+LPS+zVAD group were obviously higher than those in the NS+PBS group and the NC+PBS group, respectively. Laser scanning confocal microscopy showed that the trends in quantity and fluorescence intensity of p-MLKL protein expressions were consistent with the above results. The results from flow cytometry analysis and Hoechst staining showed that the rates of cell necrotic apoptosis in SESN2 siRNA+LPS+zVAD group were significantly higher than those in NS+LPS+zVAD group [flow cytometry analysis: (30.800±1.153)% vs. (20.800±1.114)%, Hoechst staining: (75.267±0.451)% vs. (46.267±3.371)%, both P < 0.05], indicating that knocking down SESN2 further exacerbated the occurrence of necroptosis. The necrotic apoptosis rates in SESN2 LV-RNA+LPS+zVAD group were significantly lower than those in NC+LPS+zVAD group [flow cytometry analysis: (7.160±0.669)% vs. (19.240±2.322)%, Hoechst staining: (32.433±3.113)% vs. (48.567±4.128)%, both P < 0.05], indicating that overexpressing SESN2 reversed such response and markedly reduced the proportion of necroptotic cells compared to the corresponding empty vector group. Conclusion:SESN2 exhibits an inhibitory effect on necroptosis of DC in sepsis. Targeted SESN2 expression may regulate the process of DC-mediated immune response in sepsis.

Objective:To explore the optimum time of ambulation of atrial fibrillation patients after radiofrequency ablation, to provide basis for patients' early postoperative rehabilitation.Methods:By convenient sampling method, a total of 120 patients with atrial fibrillation after radiofrequency ablation were collected at Yanghu Branch and City Branch of Changzhou Second People's Hospital from January 2020 to May 2021. They were divided into the early group, middle group and late group according to the random number table method, each group were 40 cases. All patients received routine postoperative intervention, the time of ambulation were 4, 6 and 12 h after operation in the early group, middle group and late group, respectively. The complication rate within 24 h after operation was compared among the three groups, and the comfort level of the three groups at 24, 48 and 72 h after operation was evaluated with Comfort Status Scale (GCQ).Results:Finally, 111 patients were included, including 37 in the early group, 38 in the middle group and 36 in the late group. There was no significant difference in the incidence of bleeding or hematoma, urinary retention, lumbago within 24 h after operation among the three groups ( P>0.05). The incidence of postural hypotension within 24 h after operation in the early group was 2.7% (1/37), which was lower than 21.1% (7/38) and 25.0% (9/36) in the middle and late groups, with a statistically significant difference ( χ2=4.86, 7.67, both P<0.05). At 48 and 72 h after operation, the scores of physiological dimension, psychological dimension and the total score of GCQ in the early group were (20.68 ± 3.07), (22.54 ± 3.35), (81.68 ± 6.11) and (22.54 ± 3.73), (24.38 ± 2.49), (84.92 ± 6.37), higher than those in the middle group (19.16 ± 2.19), (21.32 ± 2.27), (78.24 ± 5.58), (20.93 ± 2.85), (22.32 ± 2.04), (81.66 ± 6.56), and those in the late group (18.44 ± 1.50) (21.31 ± 1.99), (78.06 ± 4.32), (20.89 ± 2.25), (21.58 ± 1.86), (80.28 ± 6.44), the differences were statistically significant ( t values were 2.19-4.15, all P<0.05). Conclusions:Ambulation at 4 h after operation does not increase peripheral vascular complications, but can reduce the incidence of postural hypotension and improve the comfort of patients with atrial fibrillation after radiofrequency ablation.

Adult ; Aged ; Aged, 80 and over ; Agranulocytosis ; chemically induced ; Azacitidine ; adverse effects ; analogs & derivatives ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; drug therapy ; Neutrophils ; drug effects ; Young Adult

OBJECTIVETo evaluate the therapeutic efficacy and related risk factors of acute myelogenous leukemia （AML） patients treated with unrelated cord blood transplantation （UCBT）.

METHODSA retrospective analysis was performed on the clinical data of 58 AML patients that consisted of 1 case of M0, 1 case M1, 35 cases M2, 3 cases M4, 14 cases M5, 3 cases M6, and 1 case acute mixed leukemia, respectively. Of them, 1 case AML secondary to myelodysplastic syndrome, and 36 in first complete remission （CR1）, 14 in second complele remission （CR2）, 8 in non- remission （NR）, 43 cases were refractory or high-risk patients（70.1%）. The median age was 14.5 years with the median weight of 45 kg, 49 patients received sUCBT and 9 dUCBT. All the patients conditioned with intensified myeloablative regimen and received a combination of Cyclosporine A（CsA）and mycophenolate mofetil（MMF）to prevent graft- versus- host disease（GVHD）.

RESULTS56 out of 58 patients achieved engraftment with implantation rate 96.6%. The median time of ANC≥0.5×10⁹/L was 17（12-37）days, and that of PLT≥20× 109/L 33（17-140）days respectively. 24 cases developed acute GVHD（aGVHD）, the incidence rate of grade Ⅱ to Ⅳ aGVHD was 30.4%. The chronic GVHD（cGVHD）was occured in 7 patients of the 49 evaluable patients, all were limited. The estimated 3-year overall survival（OS）and disease-free survival （DFS）were（60.3±6.4）% and（60.1±6.5）% respectively. And the cumulative incidences of 3-year nonrelapse mortality（NRM）and relapse were 33.3% and 9.1% respectively. The 3- year OS rates of AML patients were（66.0 ± 6.7）% for CR and（25.0 ± 15.3）% for NR, differences were statistical significance.

CONCLUSIONFor AML patients, UCBT was conducive to improve outcome with lower incidences of cGVHD and relapse, the patients after transplantation could obtain high quality of life.

Acute Disease ; Adolescent ; Cyclosporine ; Disease-Free Survival ; Fetal Blood ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Incidence ; Leukemia, Myeloid, Acute ; Mycophenolic Acid ; analogs & derivatives ; Quality of Life ; Recurrence ; Remission Induction ; Retrospective Studies