Objective:To evaluate the effect of down-regulation of Sp1 expression on the radiosensitivity of glioma cells.Methods:The oligonucleotide sequence encoding shRNA was designed and synthesized, and cloned into LV3 (H1/GFP & Puro) vector to construct the recombinant. U251 and U87 cells were infected with recombinant lentivirus, then the stably-transfected cell lines were obtained by puromycin screening. The expression levels of Sp1 mRNA and protein were detected by RT-PCR and Western blot. Cell survival was detected by clonal survival assay, cell cycle was determined by flow cytometry, and DNA damage was measured by immunofluorescence assay, respectively.Results:At 72 h after infection, high expression of Sp1 lentiviral vector was observed in two cell lines under fluorescence microscope. RT-PCR and Western blot confirmed that the expression levels of Sp1 mRNA and protein were significantly down-regulated in both transfected cells (both P<0.01) and the silencing rates of Sp1 were above 90%. The sensitization enhancement ratio (SER) of shRNA-U251 and shRNA-U87 cells at 10% cell survival level were 1.39 and 1.18, respectively. After irradiation, the G 2/M phase ratio and the number of γ-H2AX foci in two Sp1 knockout groups were significantly increased. Conclusion:shRNA silencing of Sp1 increases the G 2/M phase arrest induced by X-ray, aggravates the degree of DNA double-strand breaks, and improves the radiosensitivity of glioma cells.

Objective:To evaluate the effect of donor age factor on intraoperative and early postoperative complications in the patients undergoing orthotopic liver transplantation (OLT).Methods:The clinical data of patients underwent OLT from January 2016 to December 2019 were retrospectively collected.The patients were divided into elderly donor group (age ≥65 yr) and non-elderly donor group (age<65 yr) according to the donor age.The preoperative and intraoperative clinical data of the two groups were matched by 1∶2 propensity score matching.The primary outcome measures were intraoperative postreperfusion syndrome and acute kidney injury and postoperative pulmonary complications within 7 days after surgery.The secondary outcome measures were time of intensive care unit stay, assisted mechanical ventilation time, postoperative length of hospital stay, rate of reintubation, and 6-month and 1-year survival and recurrence-free survival after surgery.Results:After propensity score matching was performed, 66 patients were enrolled, including 22 in elderly donor group and 44 in non-elderly donor group.There was no significant difference in the incidence of intraoperative postreperfusion syndrome, acute kidney injury and postoperative pulmonary complications within 7 days after operation, time of intensive care unit stay, assisted mechanical ventilation time, postoperative length of stay, 6-month and 1-year survival and recurrence-free survival rate between elderly donor group and non-elderly donor group ( P>0.05). Conclusion:Age factors do not exert effect on the incidence of intraoperative and early postoperative complications in the patients undergoing OLT.

Objective:To explore the effects of donor, basic recipient characteristics and intraoperative findings before orthotopic liver transplantation(OLT)on postoperative pulmonary complications and the influence of postoperative pulmonary complications on postoperative outcomes and long-term survival of patients.Methods:From January 2016 to December 2019, clinical data were retrospectively reviewed for 313 OLT recipients at First Affiliated Hospital, Nanjing Medical University.According to clinical manifestations, laboratory parameters and imaging findings within 7 days post-operation, they were divided into two groups of with and without postoperative pulmonary complications.Univariate and multivariate Logistic regression analyses were performed for general parameters of donors/recipients and perioperative variables of recipients in two groups.And Kaplan-Meier survival analysis was conducted for postoperative survival of two groups.Results:Multiariate Logistic regression analysis indicated that age, concurrent chronic diseases, low preoperative platelet count, intraoperative massive bleeding(>1 500 ml)and severe hypotension after perfusion were all independent risk factors for postoperative pulmonary complications.Compared with those without postoperative pulmonary complications, intensive care unit(ICU)stay, postoperative mechanical ventilation duration and hospital stay were significantly prolonged in patients with postoperative pulmonary complications.And rate of re-intubation, incidence of acute kidney injury and hospital mortality spiked markedly( P<0.05). Among OLT patients with benign liver diseases, overall survival rate of patients without postoperative pulmonary complications was significantly better than that of those with postoperative pulmonary complications( P<0.05). Conclusions:Age, concurrent chronic diseases, low preoperative platelet count, massive intraoperative bleeding(>1 500 ml)and severe hypotension after perfusion boost the risks of early postoperative pulmonary complications and adversely affect the outcomes of OLT recipients.

Objective    To investigate the expression and biological function of centromere protein F (CENPF) in non-small cell lung cancer (NSCLC) and the association with prognosis. Methods    Through retrieving and analyzing the bioinformatics data such as Oncomine database, Human Protein Atlas (HPA), Kaplan-Meier Plotter, STRING and DAVID database, the expression of CENPF in both normal tissues and cancer tissues of lung cancer patients was identified, and the protein interaction network analysis, functional annotation and pathway analysis of CENPF with its associated genes were carried out. Results    CENPF was overexpressed in lung adenocarcinoma tissues, but not in normal tissues. The median overall survival (OS) of NSCLC patients with low expression of CENPF was significantly longer than that of patients with high expression of CENPF. Further sub-analysis showed that low expression group from lung adenocarcinoma patients had longer median disease-free survival and OS compared with high expression group patients. CENPF and its associated hub genes mainly affected the protein K11-linked ubiquitination in biological process, anaphase-promoting complex (APC) in cell composition, ATP binding in molecular function, and cell cycle in KEGG pathway. Conclusion    CENPF is regulated in tumorigenesis and progression of NSCLC, and its protein expression level has the value of early diagnosis and prognosis evaluation in lung adenocarcinoma. It is suggested that CENPF gene can be a potential target for molecular targeted therapy of NSCLC.