1.Effect of shRNA silencing of Sp1 on radiosensitivity of glioma cells
Qiqi WANG ; Jiayu ZENG ; Qiang LI ; Xiongxiong LIU
Chinese Journal of Radiation Oncology 2022;31(7):638-642
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.
2.Effect of donor age factor on intraoperative and early postoperative complications in patients undergoing orthotopic liver transplantation
Zhiying ZHENG ; Hui ZHANG ; Xinyang LIU ; Xiongxiong PAN
Chinese Journal of Anesthesiology 2021;41(8):919-923
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.
3.Analysis of risk factors of early pulmonary complications after orthotopic liver transplantation: a report of 313 cases
Xinyang LIU ; Hui ZHANG ; Zhiying ZHENG ; Xiongxiong PAN
Chinese Journal of Organ Transplantation 2022;43(3):141-145
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.
4.Effect of dexmedetomidine on the intraoperative and early postoperative complications of patients undergoing orthotopic liver transplantation
Junwei KANG ; Hui ZHANG ; Yining CHEN ; Zhiying ZHENG ; Xinyang LIU ; Xiongxiong PAN
Chinese Journal of Surgery 2024;62(2):155-161
Objective:To investigate the effect of dexmedetomidine on the intraoperative and early postoperative complications of patients undergoing orthotopic liver transplantation.Methods:This is a retrospective cohort study. The clinical data of 399 patients who underwent orthotopic liver transplantation at the First Affiliated Hospital of Nanjing Medical University from January 2016 to September 2020 were retrospectively collected. There were 319 males and 80 females, aged (50.9±10.2) years (range: 10 to 73 years). These patients were divided into the control group (369 cases) and the dexmedetomidine group (30 cases) according to whether dexmedetomidine was continuously pumped intravenously during the operation until the operation ended. The 1∶2 propensity score matching was used to match the preoperative and intraoperative conditions of the two groups of patients, and the caliper width was 0.2. Outcome indicators included intraoperative postreperfusion syndrome, acute kidney injury and pulmonary complications within 7 days after surgery, length of hospital stay, time of stay in ICU, duration of assisted mechanical ventilation, rate of reintubation, 6-month and 1-year survival and recurrence-free survival rate after surgery. The independent sample t test, χ2 test, Mann-Whitney U test or Fisher exact test was used to statistically analyze the data of the two groups of patients, respectively. Survival curves of overall survival and disease-free-survival were plotted by Kaplan-Meier method, and the survival rate and recurrence-free survival rate were compared by Log-rank test. Results:A total of 78 patients were included after propensity score matching, including 26 in the dexmedetomidine group and 52 in the control group. The incidence of acute kidney injury in the dexmedetomidine group within 7 days after surgery was 0 (0/26), significantly lower than that of the control group (21.2%,11/52)(corrected χ2=4.776, P=0.029). There were no significant differences in the incidence of intraoperative postreperfusion syndrome and pulmonary complications within 7 days after surgery, length of hospital stay, ICU time, the duration of assisted mechanical ventilation, rate of reintubation, 6-month and 1-year survival, and recurrence-free survival rate after surgery between the two groups (all P>0.05). Conclusion:Continuous infusion of dexmedetomidine via intravenous pump during operation may be beneficial in reducing the incidence of acute kidney injury within 7 days after orthotopic liver transplantation.
5.Effect of dexmedetomidine on the intraoperative and early postoperative complications of patients undergoing orthotopic liver transplantation
Junwei KANG ; Hui ZHANG ; Yining CHEN ; Zhiying ZHENG ; Xinyang LIU ; Xiongxiong PAN
Chinese Journal of Surgery 2024;62(2):155-161
Objective:To investigate the effect of dexmedetomidine on the intraoperative and early postoperative complications of patients undergoing orthotopic liver transplantation.Methods:This is a retrospective cohort study. The clinical data of 399 patients who underwent orthotopic liver transplantation at the First Affiliated Hospital of Nanjing Medical University from January 2016 to September 2020 were retrospectively collected. There were 319 males and 80 females, aged (50.9±10.2) years (range: 10 to 73 years). These patients were divided into the control group (369 cases) and the dexmedetomidine group (30 cases) according to whether dexmedetomidine was continuously pumped intravenously during the operation until the operation ended. The 1∶2 propensity score matching was used to match the preoperative and intraoperative conditions of the two groups of patients, and the caliper width was 0.2. Outcome indicators included intraoperative postreperfusion syndrome, acute kidney injury and pulmonary complications within 7 days after surgery, length of hospital stay, time of stay in ICU, duration of assisted mechanical ventilation, rate of reintubation, 6-month and 1-year survival and recurrence-free survival rate after surgery. The independent sample t test, χ2 test, Mann-Whitney U test or Fisher exact test was used to statistically analyze the data of the two groups of patients, respectively. Survival curves of overall survival and disease-free-survival were plotted by Kaplan-Meier method, and the survival rate and recurrence-free survival rate were compared by Log-rank test. Results:A total of 78 patients were included after propensity score matching, including 26 in the dexmedetomidine group and 52 in the control group. The incidence of acute kidney injury in the dexmedetomidine group within 7 days after surgery was 0 (0/26), significantly lower than that of the control group (21.2%,11/52)(corrected χ2=4.776, P=0.029). There were no significant differences in the incidence of intraoperative postreperfusion syndrome and pulmonary complications within 7 days after surgery, length of hospital stay, ICU time, the duration of assisted mechanical ventilation, rate of reintubation, 6-month and 1-year survival, and recurrence-free survival rate after surgery between the two groups (all P>0.05). Conclusion:Continuous infusion of dexmedetomidine via intravenous pump during operation may be beneficial in reducing the incidence of acute kidney injury within 7 days after orthotopic liver transplantation.
6.Analysis of risk factors and prognosis for early acute kidney injury after orthotopic liver trans-plantation
Yining CHEN ; Hui ZHANG ; Junwei KANG ; Zhiying ZHENG ; Xinyang LIU ; Xiongxiong PAN
Chinese Journal of Digestive Surgery 2024;23(7):952-960
Objective:To analyze the risk factors and prognosis for early acute kidney injury (AKI) after orthotopic liver transplantation (OLT).Methods:The retrospective study was conduc-ted. The clinicopathological data of 340 pairs of donor and recipients undergoing OLT in The First Affiliated Hospital of Nanjing Medical University from January 2016 to January 2020 were collected. There were 262 males and 78 females of donors. There were 268 males and 72 females of recipients, aged (51±11)years. Of 340 recipients, 217 cases without postoperative early AKI were divided into the non-AKI group and 123 cases with postoperative early AKI were divided into the AKI group. Measure-ment data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distri-bution were represented as M( IQR), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data was conducted using the non-parameter test. Multivariate analysis was conducted using the binary Logistic regre-ssion model with forward method. The nomogram predictive model was constructed using the R software with its RMS package (R3.6.1). The efficacy of the predictive model was validated using the area under curve (AUC) of the receiver operating characteristic (ROC) curve, and internal validation of the predictive model was performed using the Bootstrap method. The Kaplan-Meier method was used to draw survival curves, and Log-rank test was used for survival analysis. Results:(1) Com-parison of preoperative clinical characteristics between donors and recipients of the non-AKI group and the AKI group. There was a significant difference in overweight of donors between the non-AKI group and the AKI group ( P<0.05). There were significant differences in preoperative hypertension, viral hepatitis, pathological types, international normalized ratio, fibrinogen levels, platelet (PLT), hemoglobin, and anemia of recipients between the non-AKI group and the AKI group ( P<0.05). (2) Comparison of surgical situations between recipients of the non-AKI group and the AKI group. There were significant differences in intraoperative urine output, volume of intraoperative blood loss, peak serum potassium after reperfusion, massive transfusion, plasma infusion, cryoprecipitate infusion, and aminocaproic acid use of recipients between the non-AKI group and the AKI group ( P<0.05). (3) Influencing factors for postoperative early AKI and construction and evaluation of the nomogram predictive model for postoperative early AKI. Results of multivariate analysis showed that donors of overweight, recipients of preoperative hypertension, recipients of non-viral hepatitis, recipients of preoperative severe PLT reduction, recipients of less intraoperative urine output, recipients of severe post-reperfusion hypotension, recipients of high peak serum potassium after reperfusion, recipients with intraoperative plasma infusion were independent risk factors for postoperative early AKI ( odds ratio=1.982, 3.365, 0.519, 3.615, 0.169, 2.480, 1.500, 1.001, 95% confidence interval as 1.160-3.388, 1.649-6.865, 0.293-0.917, 1.358-9.621, 0.061-0.464, 1.246-4.934, 1.003-2.243, 1.000-1.001, P<0.05). The nomogram predictive model for postoperative early AKI was constructed based on the results of multivariate analysis. Results of ROC curve showed the AUC was 0.769 (95% confidence interval as 0.717-0.820). Results of the calibration curve showed that the predictive results of nomogram predictive model fitted well with the actual situation, with a mean absolute error of 0.016. (4) Comparison of prognosis between recipients of the non-AKI group and the AKI group. There were significant differences in postopera-tive peak creatinine, peak brain natriuretic peptide, duration of intensive care unit stay, mechanical ventilation time, re-intubation of recipients between the non-AKI group and the AKI group ( Z=-4.836, -5.652, -5.861, -6.533, χ2=14.676, P<0.05). All 340 recipients were followed up. For recipients of hepatocellular carcinoma, the 6-month survival rates after surgery were 87.8% and 75.6% of the non-AKI group and the AKI group, respectively, showing a significant difference between them ( χ2=4.010, P<0.05), and the overall survival rates were 46.7% and 56.1% of the non-AKI group and the AKI group, respectively, showing no significant difference between them ( χ2=0.047, P>0.05). For recipients of benign liver disease, the 6-month survival rates after surgery were 89.8% and 78.0% of the non-AKI group and the AKI group, respectively, showing a significant difference between them ( χ2=6.401, P<0.05), and the overall survival rates were 81.4% and 68.0% of the non-AKI group and the AKI group, respectively, showing a significant difference between them ( χ2=4.452, P<0.05). Conclusions:Donors of overweight, recipients of preoperative hypertension, recipients of non-viral hepatitis, recipients of preoperative severe PLT reduction, reci-pients of less intraoperative urine output, recipients of severe post-reperfusion hypotension, recipi-ents of high peak serum potassium after reperfusion, recipients with intraoperative plasma transfu-sion were independent risk factors for postoperative early AKI. Nomogram predictive model has well clinical application value. For recipients of benign liver disease, the 6-month survival rate after surgery and overall survival rate of recipients in the non-AKI group are superior to those of the AKI group.
7.Effects of centromere protein F expression on biological behavior and prognosis of non-small cell lung cancer
Yanxiang CHANG ; Lei JIN ; Xiongxiong BAI ; Hao LIU ; Shuonan XU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2022;29(06):711-716
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.