1.Effect of VEGF on the expression of genes related to ovarian steroid synthesis in mice and its mechanism
Zhi-Hui ZHANG ; Hong-Xia GAO ; Guo-Qing WANG ; Wei HOU ; Chang ZOU ; Xiao-Dan LU
Medical Journal of Chinese People's Liberation Army 2024;49(6):679-685
		                        		
		                        			
		                        			Objective To investigate the effect of vascular endothelial growth factor(VEGF)on the expression of genes related to ovarian steroid synthesis in mice and its underlying mechanism.Methods A transgenic mouse model with tetracycline-reversible regulation of VEGF expression was used,and the genotype of mice was identified by polymerase chain reaction(PCR).Twenty mice were divided into normal VEGF expression group(Dox+,n=10)and VEGF expression inhibition group(Dox-,n=10)by feeding them doxycycline.Western blotting was used to detect the expression of VEGF protein in ovarian tissues.Fluorescence quantitative PCR was used to detect the mRNA expression of VEGF,KDR and genes known to play roles in follicle development,such as follicle-stimulating hormone(FSH)and inhibin B(INHBB).HE staining was used to observe changes in ovarian tissue.Total RNA was extracted from mouse ovarian tissues for transcriptome sequencing,and the relevant differential genes were analyzed by FPKM and log2FC values.Results Compared with the Dox+group,the mRNA and protein levels of VEGF in the Dox-group significantly reduced,and the mRNA levels of KDR also significantly decreased(P<0.05).HE staining results showed that compared with the Dox+group,follicular development was impaired and atresia follicles appeared in the Dox-group.Sequencing analysis identified that significant differences in follicular development-related genes and steroid synthesis-related genes between the two groups(P<0.05).Enrichment analysis showed that VEGF in mouse ovaries mainly regulates ovarian steroidogenesis and other pathways.Fluorescence quantitative PCR results demonstrated that compared with the Dox+group,the follicular development-related genes(INHBB and FSHR)in the ovarian tissues of the Dox-group were significantly up-regulated(P<0.05),whereas the key genes of steroid synthesis(StAR,CYP11A1,3β-HSD)were significantly down-regulated(P<0.05).The quantitative results were basically consistent with the sequencing results.Conclusion Mice with inhibited VEGF exhibited ovarian follicular dysplasia,potentially due to the mechanism whereby VEGF inhibition downregulated the expression of genes associated with steroid synthesis,such as FSH and INHBB,thereby obstructing cholesterol metabolism.
		                        		
		                        		
		                        		
		                        	
2.Epidemiological characteristics of a 2019-nCoV outbreak caused by Omicron variant BF.7 in Shenzhen.
Yan Peng CHENG ; Dong Feng KONG ; Jia ZHANG ; Zi Quan LYU ; Zhi Gao CHEN ; Hua Wei XIONG ; Yan LU ; Qing Shan LUO ; Qiu Ying LYU ; Jin ZHAO ; Ying WEN ; Jia WAN ; Fang Fang LU ; Jian Hua LU ; Xuan ZOU ; Zhen ZHANG
Chinese Journal of Epidemiology 2023;44(3):379-385
		                        		
		                        			
		                        			Objective: To explore the epidemiological characteristic of a COVID-19 outbreak caused by 2019-nCoV Omicron variant BF.7 and other provinces imported in Shenzhen and analyze transmission chains and characteristics. Methods: Field epidemiological survey was conducted to identify the transmission chain, analyze the generation relationship among the cases. The 2019-nCoV nucleic acid positive samples were used for gene sequencing. Results: From 8 to 23 October, 2022, a total of 196 cases of COVID-19 were reported in Shenzhen, all the cases had epidemiological links. In the cases, 100 were men and 96 were women, with a median of age, M (Q1, Q3) was 33(25, 46) years. The outbreak was caused by traverlers initial cases infected with 2019-nCoV who returned to Shenzhen after traveling outside of Guangdong Province.There were four transmission chains, including the transmission in place of residence and neighbourhood, affecting 8 persons, transmission in social activity in the evening on 7 October, affecting 65 persons, transmission in work place on 8 October, affecting 48 persons, and transmission in a building near the work place, affecting 74 persons. The median of the incubation period of the infection, M (Q1, Q3) was 1.44 (1.11, 2.17) days. The incubation period of indoor exposure less than that of the outdoor exposure, M (Q1, Q3) was 1.38 (1.06, 1.84) and 1.95 (1.22, 2.99) days, respcetively (Wald χ2=10.27, P=0.001). With the increase of case generation, the number and probability of gene mutation increased. In the same transmission chain, the proportion of having 1-3 mutation sites was high in the cases in the first generation. Conclusions: The transmission chains were clear in this epidemic. The incubation period of Omicron variant BF.7 infection was shorter, the transmission speed was faster, and the gene mutation rate was higher. It is necessary to conduct prompt response and strict disease control when epidemic occurs.
		                        		
		                        		
		                        		
		                        			Male
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			SARS-CoV-2
		                        			;
		                        		
		                        			COVID-19/epidemiology*
		                        			;
		                        		
		                        			Disease Outbreaks
		                        			;
		                        		
		                        			Epidemics
		                        			;
		                        		
		                        			China/epidemiology*
		                        			
		                        		
		                        	
3.Efficacy of partial nephrectomy in patients with localized renal carcinoma: a 20-year experience of 2 046 patients in a single center.
Xiang Peng ZOU ; Kang NING ; Zhi Ling ZHANG ; Long Bin XIONG ; Yu Lu PENG ; Zhao Hui ZHOU ; Yi Xin HUANG ; Xin LUO ; Ji Bin LI ; Pei DONG ; Sheng Jie GUO ; Hui HAN ; Fang Jian ZHOU
Chinese Journal of Surgery 2023;61(5):395-402
		                        		
		                        			
		                        			Objectives: To analyze the long-term survival of patients with localized renal cell carcinoma after partical nephrectomy. Methods: The clinicopathological records and survival follow-up data of 2 046 patients with localized renal cell carcinoma, who were treated with partial nephrectomy from August 2001 to February 2021 in the Department of Urology, Sun Yat-sen University Cancer Center, were retrospectively analyzed. There were 1 402 males and 644 females, aged (M(IQR)) 51 (19) years (range: 6 to 86 years). The primary end point of this study was cancer-specific survival. Survival curves were estimated using the Kaplan-Meier method, and the difference test was performed by Log-rank test. Univariate and multivariate Cox analysis were fitted to determine factors associated with cancer-specific survival. Results: The follow-up time was 49.2 (48.0) months (range: 1 to 229 months), with 1 974 patients surviving and 72 dying. The median cancer-specific survival time has not yet been reached. The 5- and 10-year cancer specific survival rates were 97.0% and 91.2%, respectively. The 10-year cancer-specific survival rates for stage pT1a (n=1 447), pT1b (n=523) and pT2 (n=58) were 95.3%, 81.8%, and 81.7%, respectively. The 10-year cancer-specific survival rates of patients with nuclear grade 1 (n=226), 2 (n=1 244) and 3 to 4 (n=278) were 96.6%, 89.4%, and 85.5%, respectively. There were no significant differences in 5-year cancer-specific survival rates among patients underwent open, laparoscopic, or robotic surgery (96.7% vs. 97.1% vs. 97.5%, P=0.600). Multivariate analysis showed that age≥50 years (HR=3.93, 95%CI: 1.82 to 8.47, P<0.01), T stage (T1b vs. T1a: HR=3.31, 95%CI: 1.83 to 5.99, P<0.01; T2+T3 vs. T1a: HR=2.88, 95%CI: 1.00 to 8.28, P=0.049) and nuclear grade (G3 to 4 vs. G1: HR=2.81, 95%CI: 1.01 to 7.82, P=0.048) were independent prognostic factors of localized renal cell carcinoma after partial nephrectomy. Conclusions: The long-term cancer-specific survival rates of patients with localized renal cancer after partial nephrectomy are satisfactory. The type of operation (open, laparoscopic, or robotic) has no significant effect on survival. However, patients with older age, higher nuclear grade, and higher T stage have a lower cancer-specific survival rate. Grasping surgical indications, attaching importance to preoperative evaluation, perioperative management, and postoperative follow-up, could benefit achieving satisfactory long-term survival.
		                        		
		                        		
		                        		
		                        	
4.Establishment and validation of a novel nomogram to predict overall survival after radical nephrectomy.
Long Bin XIONG ; Xiang Peng ZOU ; Kang NING ; Xin LUO ; Yu Lu PENG ; Zhao Hui ZHOU ; Jun WANG ; Zhen LI ; Chun Ping YU ; Pei DONG ; Sheng Jie GUO ; Hui HAN ; Fang Jian ZHOU ; Zhi Ling ZHANG
Chinese Journal of Oncology 2023;45(8):681-689
		                        		
		                        			
		                        			Objective: To establish a nomogram prognostic model for predicting the 5-, 10-, and 15-year overall survival (OS) of non-metastatic renal cell carcinoma patients managed with radical nephrectomy (RN), compare the modelled results with the results of pure pathologic staging, the Karakiewicz nomogram and the Mayo Clinic Stage, Size, Grade, and Necrosis (SSIGN) score commonly used in foreign countries, and stratify the patients into different prognostic risk subgroups. Methods: A total of 1 246 non-metastatic renal cell carcinoma patients managed with RN in Sun Yat-sen University Cancer Center (SYSUCC) from 1999 to 2020 were retrospectively analyzed. Multivariate Cox regression analysis was used to screen the variables that influence the prognosis for nomogram establishment, and the bootstrap random sampling was used for internal validation. The time-receiver operating characteristic curve (ROC), the calibration curve and the clinical decision curve analysis (DCA) were applied to evaluate the nomogram. The prediction efficacy of the nomogram and that of the pure pathologic staging, the Karakiewicz nomogram and the SSIGN score was compared through the area under the curve (AUC). Finally, patients were stratified into different risk subgroups according to our nomogram scores. Results: A total of 1 246 patients managed with RN were enrolled in this study. Multivariate Cox regression analysis showed that age, smoking history, pathological nuclear grade, sarcomatoid differentiation, tumor necrosis and pathological T and N stages were independent prognostic factors for RN patients (all P<0.05). A nomogram model named SYSUCC based on these factors was built to predict the 5-, 10-, and 15-year survival rate of the participating patients. In the bootstrap random sampling with 1 000 iterations, all these factors occurred for more than 800 times as independent predictors. The Harrell's concordance index (C-index) of SYSUCC was higher compared with pure pathological staging [0.770 (95% CI: 0.716-0.823) vs 0.674 (95% CI: 0.621-0.728)]. The calibration curve showed that the survival rate as predicted by the SYSUCC model simulated the actual rate, while the clinical DCA showed that the SYSUCC nomogram has a benefit in certain probability ranges. In the ROC analysis that included 857 patients with detailed pathological nuclear stages, the nomogram had a larger AUC (5-/10-year AUC: 0.823/0.804) and better discriminating ability than pure pathological staging (5-/10-year AUC: 0.701/0.658), Karakiewicz nomogram (5-/10-year AUC: 0.772/0.734) and SSIGN score (5-/10-year AUC: 0.792/0.750) in predicting the 5-/10-year OS of RN patients (all P<0.05). In addition, the AUC of the SYSUCC nomogram for predicting the 15-year OS (0.820) was larger than that of the SSIGN score (0.709), and there was no statistical difference (P<0.05) between the SYSUCC nomogram, pure pathological staging (0.773) and the Karakiewicz nomogram (0.826). The calibration curve was close to the standard curve, which indicated that the model has good predictive performance. Finally, patients were stratified into low-, intermediate-, and high-risk subgroups (738, 379 and 129, respectively) according to the SYSUCC nomogram scores, among whom patients in intermediate- and high-risk subgroups had a worse OS than patients in the low-risk subgroup (intermediate-risk group vs. low-risk group: HR=4.33, 95% CI: 3.22-5.81, P<0.001; high-risk group vs low-risk group: HR=11.95, 95% CI: 8.29-17.24, P<0.001), and the high-risk subgroup had a worse OS than the intermediate-risk group (HR=2.63, 95% CI: 1.88-3.68, P<0.001). Conclusions: Age, smoking history, pathological nuclear grade, sarcomatoid differentiation, tumor necrosis and pathological stage were independent prognostic factors for non-metastasis renal cell carcinoma patients after RN. The SYSUCC nomogram based on these independent prognostic factors can better predict the 5-, 10-, and 15-year OS than pure pathological staging, the Karakiewicz nomogram and the SSIGN score of patients after RN. In addition, the SYSUCC nomogram has good discrimination, agreement, risk stratification and clinical application potential.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Nomograms
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Carcinoma, Renal Cell/pathology*
		                        			;
		                        		
		                        			Prognosis
		                        			;
		                        		
		                        			Risk Factors
		                        			;
		                        		
		                        			Nephrectomy
		                        			;
		                        		
		                        			Kidney Neoplasms/pathology*
		                        			;
		                        		
		                        			Necrosis
		                        			
		                        		
		                        	
5.Long-term survival analysis of 1 367 patients treated with radical nephrectomy from a single center.
Xiang Peng ZOU ; Kang NING ; Zhi Ling ZHANG ; Ling ZOU ; Long Bin XIONG ; Yu Lu PENG ; Zhao Hui ZHOU ; Hui Ming LIU ; Chun Ping YU ; Pei DONG ; Sheng Jie GUO ; Hui HAN ; Fang Jian ZHOU
Chinese Journal of Oncology 2023;45(11):981-987
		                        		
		                        			
		                        			Objective: To report the long-term survival of renal cell carcinoma (RCC) patients treated with radical nephrectomy in Sun Yat-sen University Cancer Center. Methods: We retrospectively analyzed the clinical, pathological and follow-up records of 1 367 non-metastatic RCC patients treated with radical nephrectomy from 1999 to 2020 in this center. The primary endpoint of this study was overall survival rate. Survival curves were estimated using the Kaplan-Meier method, and group differences were compared through Log-rank test. Univariate and multivariate Cox analysis were fit to determine the clinical and pathological features associated with overall survival rate. Results: A total of 1 367 patients treated with radical nephrectomy with complete follow-up data were included in the study. The median follow-up time was 52.6 months, and 1 100 patients survived and 267 died, with the median time to overall survival not yet reached. The 5-year and 10-year overall survival rates were 82.8% and 74.9%, respectively. The 5-year and 10-year overall survival rates of Leibovich low-risk patients were 93.3% and 88.2%, respectively; of Leibovich intermediate-risk patients were 82.2% and 72.3%, respectively; and of Leibovich high-risk patients were 50.5% and 30.2%, respectively. There were significant differences in the long-term survival among the three groups (P<0.001). The 10-year overall survival rates for patients with pT1, pT2, pT3 and pT4 RCC were 83.2%, 73.6%, 55.0% and 31.4%, respectively. There were significant differences among pT1, pT2, pT3 and pT4 patients(P<0.001). The 5-year and 10-year overall survival rates of patients with lymph node metastasis were 48.5% and 35.6%, respectively, and those of patients without lymph node metastasis were 85.1% and 77.5%, respectively. There was significant difference in the long-term survival between patients with lymph node metastasis and without lymph node metastasis. The 10-year overall survival rate was 96.2% for nuclear Grade 1, 81.6% for nuclear Grade 2, 60.5% for nuclear Grade 3, and 43.4% for nuclear Grade 4 patients. The difference was statistically significant. There was no significant difference in the long-term survival between patients with localized renal cancer (pT1-2N0M0) who underwent open surgery and minimally invasive surgery (10-year overall survival rate 80.5% vs 85.6%, P=0.160). Multivariate Cox analysis showed that age≥55 years (HR=2.11, 95% CI: 1.50-2.96, P<0.001), T stage(T3+ T4 vs T1a: HR=2.37, 95% CI: 1.26-4.46, P=0.008), local lymph node metastasis (HR=3.04, 95%CI: 1.81-5.09, P<0.001), nuclear grade (G3-G4 vs G1: HR=4.21, 95%CI: 1.51-11.75, P=0.006), tumor necrosis (HR=1.66, 95% CI: 1.17-2.37, P=0.005), sarcomatoid differentiation (HR=2.39, 95% CI: 1.31-4.35, P=0.005) and BMI≥24kg/m(2) (HR=0.56, 95%CI: 0.39-0.80, P=0.001) were independent factors affecting long-term survival after radical nephrectomy. Conclusions: The long-term survival of radical nephrectomy in patients with renal cell carcinoma is satisfactory. Advanced age, higher pathological stage and grade, tumor necrosis and sarcomatoid differentiation were the main adverse factors affecting the prognosis of patients. Higher body mass index was a protective factor for the prognosis of patients.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Middle Aged
		                        			;
		                        		
		                        			Carcinoma, Renal Cell/secondary*
		                        			;
		                        		
		                        			Lymphatic Metastasis
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Neoplasm Staging
		                        			;
		                        		
		                        			Kidney Neoplasms/pathology*
		                        			;
		                        		
		                        			Prognosis
		                        			;
		                        		
		                        			Nephrectomy
		                        			;
		                        		
		                        			Survival Analysis
		                        			;
		                        		
		                        			Necrosis/surgery*
		                        			;
		                        		
		                        			Survival Rate
		                        			
		                        		
		                        	
6.Establishment and validation of a novel nomogram to predict overall survival after radical nephrectomy.
Long Bin XIONG ; Xiang Peng ZOU ; Kang NING ; Xin LUO ; Yu Lu PENG ; Zhao Hui ZHOU ; Jun WANG ; Zhen LI ; Chun Ping YU ; Pei DONG ; Sheng Jie GUO ; Hui HAN ; Fang Jian ZHOU ; Zhi Ling ZHANG
Chinese Journal of Oncology 2023;45(8):681-689
		                        		
		                        			
		                        			Objective: To establish a nomogram prognostic model for predicting the 5-, 10-, and 15-year overall survival (OS) of non-metastatic renal cell carcinoma patients managed with radical nephrectomy (RN), compare the modelled results with the results of pure pathologic staging, the Karakiewicz nomogram and the Mayo Clinic Stage, Size, Grade, and Necrosis (SSIGN) score commonly used in foreign countries, and stratify the patients into different prognostic risk subgroups. Methods: A total of 1 246 non-metastatic renal cell carcinoma patients managed with RN in Sun Yat-sen University Cancer Center (SYSUCC) from 1999 to 2020 were retrospectively analyzed. Multivariate Cox regression analysis was used to screen the variables that influence the prognosis for nomogram establishment, and the bootstrap random sampling was used for internal validation. The time-receiver operating characteristic curve (ROC), the calibration curve and the clinical decision curve analysis (DCA) were applied to evaluate the nomogram. The prediction efficacy of the nomogram and that of the pure pathologic staging, the Karakiewicz nomogram and the SSIGN score was compared through the area under the curve (AUC). Finally, patients were stratified into different risk subgroups according to our nomogram scores. Results: A total of 1 246 patients managed with RN were enrolled in this study. Multivariate Cox regression analysis showed that age, smoking history, pathological nuclear grade, sarcomatoid differentiation, tumor necrosis and pathological T and N stages were independent prognostic factors for RN patients (all P<0.05). A nomogram model named SYSUCC based on these factors was built to predict the 5-, 10-, and 15-year survival rate of the participating patients. In the bootstrap random sampling with 1 000 iterations, all these factors occurred for more than 800 times as independent predictors. The Harrell's concordance index (C-index) of SYSUCC was higher compared with pure pathological staging [0.770 (95% CI: 0.716-0.823) vs 0.674 (95% CI: 0.621-0.728)]. The calibration curve showed that the survival rate as predicted by the SYSUCC model simulated the actual rate, while the clinical DCA showed that the SYSUCC nomogram has a benefit in certain probability ranges. In the ROC analysis that included 857 patients with detailed pathological nuclear stages, the nomogram had a larger AUC (5-/10-year AUC: 0.823/0.804) and better discriminating ability than pure pathological staging (5-/10-year AUC: 0.701/0.658), Karakiewicz nomogram (5-/10-year AUC: 0.772/0.734) and SSIGN score (5-/10-year AUC: 0.792/0.750) in predicting the 5-/10-year OS of RN patients (all P<0.05). In addition, the AUC of the SYSUCC nomogram for predicting the 15-year OS (0.820) was larger than that of the SSIGN score (0.709), and there was no statistical difference (P<0.05) between the SYSUCC nomogram, pure pathological staging (0.773) and the Karakiewicz nomogram (0.826). The calibration curve was close to the standard curve, which indicated that the model has good predictive performance. Finally, patients were stratified into low-, intermediate-, and high-risk subgroups (738, 379 and 129, respectively) according to the SYSUCC nomogram scores, among whom patients in intermediate- and high-risk subgroups had a worse OS than patients in the low-risk subgroup (intermediate-risk group vs. low-risk group: HR=4.33, 95% CI: 3.22-5.81, P<0.001; high-risk group vs low-risk group: HR=11.95, 95% CI: 8.29-17.24, P<0.001), and the high-risk subgroup had a worse OS than the intermediate-risk group (HR=2.63, 95% CI: 1.88-3.68, P<0.001). Conclusions: Age, smoking history, pathological nuclear grade, sarcomatoid differentiation, tumor necrosis and pathological stage were independent prognostic factors for non-metastasis renal cell carcinoma patients after RN. The SYSUCC nomogram based on these independent prognostic factors can better predict the 5-, 10-, and 15-year OS than pure pathological staging, the Karakiewicz nomogram and the SSIGN score of patients after RN. In addition, the SYSUCC nomogram has good discrimination, agreement, risk stratification and clinical application potential.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Nomograms
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Carcinoma, Renal Cell/pathology*
		                        			;
		                        		
		                        			Prognosis
		                        			;
		                        		
		                        			Risk Factors
		                        			;
		                        		
		                        			Nephrectomy
		                        			;
		                        		
		                        			Kidney Neoplasms/pathology*
		                        			;
		                        		
		                        			Necrosis
		                        			
		                        		
		                        	
7.Long-term survival analysis of 1 367 patients treated with radical nephrectomy from a single center.
Xiang Peng ZOU ; Kang NING ; Zhi Ling ZHANG ; Ling ZOU ; Long Bin XIONG ; Yu Lu PENG ; Zhao Hui ZHOU ; Hui Ming LIU ; Chun Ping YU ; Pei DONG ; Sheng Jie GUO ; Hui HAN ; Fang Jian ZHOU
Chinese Journal of Oncology 2023;45(11):981-987
		                        		
		                        			
		                        			Objective: To report the long-term survival of renal cell carcinoma (RCC) patients treated with radical nephrectomy in Sun Yat-sen University Cancer Center. Methods: We retrospectively analyzed the clinical, pathological and follow-up records of 1 367 non-metastatic RCC patients treated with radical nephrectomy from 1999 to 2020 in this center. The primary endpoint of this study was overall survival rate. Survival curves were estimated using the Kaplan-Meier method, and group differences were compared through Log-rank test. Univariate and multivariate Cox analysis were fit to determine the clinical and pathological features associated with overall survival rate. Results: A total of 1 367 patients treated with radical nephrectomy with complete follow-up data were included in the study. The median follow-up time was 52.6 months, and 1 100 patients survived and 267 died, with the median time to overall survival not yet reached. The 5-year and 10-year overall survival rates were 82.8% and 74.9%, respectively. The 5-year and 10-year overall survival rates of Leibovich low-risk patients were 93.3% and 88.2%, respectively; of Leibovich intermediate-risk patients were 82.2% and 72.3%, respectively; and of Leibovich high-risk patients were 50.5% and 30.2%, respectively. There were significant differences in the long-term survival among the three groups (P<0.001). The 10-year overall survival rates for patients with pT1, pT2, pT3 and pT4 RCC were 83.2%, 73.6%, 55.0% and 31.4%, respectively. There were significant differences among pT1, pT2, pT3 and pT4 patients(P<0.001). The 5-year and 10-year overall survival rates of patients with lymph node metastasis were 48.5% and 35.6%, respectively, and those of patients without lymph node metastasis were 85.1% and 77.5%, respectively. There was significant difference in the long-term survival between patients with lymph node metastasis and without lymph node metastasis. The 10-year overall survival rate was 96.2% for nuclear Grade 1, 81.6% for nuclear Grade 2, 60.5% for nuclear Grade 3, and 43.4% for nuclear Grade 4 patients. The difference was statistically significant. There was no significant difference in the long-term survival between patients with localized renal cancer (pT1-2N0M0) who underwent open surgery and minimally invasive surgery (10-year overall survival rate 80.5% vs 85.6%, P=0.160). Multivariate Cox analysis showed that age≥55 years (HR=2.11, 95% CI: 1.50-2.96, P<0.001), T stage(T3+ T4 vs T1a: HR=2.37, 95% CI: 1.26-4.46, P=0.008), local lymph node metastasis (HR=3.04, 95%CI: 1.81-5.09, P<0.001), nuclear grade (G3-G4 vs G1: HR=4.21, 95%CI: 1.51-11.75, P=0.006), tumor necrosis (HR=1.66, 95% CI: 1.17-2.37, P=0.005), sarcomatoid differentiation (HR=2.39, 95% CI: 1.31-4.35, P=0.005) and BMI≥24kg/m(2) (HR=0.56, 95%CI: 0.39-0.80, P=0.001) were independent factors affecting long-term survival after radical nephrectomy. Conclusions: The long-term survival of radical nephrectomy in patients with renal cell carcinoma is satisfactory. Advanced age, higher pathological stage and grade, tumor necrosis and sarcomatoid differentiation were the main adverse factors affecting the prognosis of patients. Higher body mass index was a protective factor for the prognosis of patients.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Middle Aged
		                        			;
		                        		
		                        			Carcinoma, Renal Cell/secondary*
		                        			;
		                        		
		                        			Lymphatic Metastasis
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Neoplasm Staging
		                        			;
		                        		
		                        			Kidney Neoplasms/pathology*
		                        			;
		                        		
		                        			Prognosis
		                        			;
		                        		
		                        			Nephrectomy
		                        			;
		                        		
		                        			Survival Analysis
		                        			;
		                        		
		                        			Necrosis/surgery*
		                        			;
		                        		
		                        			Survival Rate
		                        			
		                        		
		                        	
8.The role and mechanism of autophagy in lipopolysaccharide-induced inflammatory response of A549 cells.
Jia SHI ; Hui-Xian TAO ; Yan GUO ; Yun-Su ZOU ; Mu-Zi WANG ; Zhi-Tao LU ; Yi-Fang DING ; Wei-Dong XU ; Xiao-Guang ZHOU
Chinese Journal of Contemporary Pediatrics 2022;24(10):1161-1170
		                        		
		                        			OBJECTIVES:
		                        			To study the role and mechanism of autophagy in lipopolysaccharide (LPS)-induced inflammatory response of human alveolar epithelial A549 cells.
		                        		
		                        			METHODS:
		                        			A549 cells were stimulated with LPS to establish a cell model of inflammatory response, and were then grouped (n=3 each) by concentration (0, 1, 5, and 10 μg/mL) and time (0, 4, 8, 12, and 24 hours). The A549 cells were treated with autophagy inhibitor 3-methyladenine (3-MA) to be divided into four groups (n=3 each): control, LPS, 3-MA, and 3-MA+LPS. The A549 cells were treated with autophagy agonist rapamycin (RAPA) to be divided into four groups (n=3 each): control, LPS, RAPA, and RAPA+LPS. The A549 cells were transfected with the Toll-like receptor 4 (TLR4) overexpression plasmid to be divided into four groups (n=3 each): TLR4 overexpression control, TLR4 overexpression, TLR4 overexpression control+LPS, and TLR4 overexpression+LPS. The A549 cells were transfected with TLR4 siRNA to be divided into four groups (n=3 each): TLR4 silencing control,TLR4 silencing, TLR4 silencing control+LPS, and TLR4 silencing+LPS. CCK-8 assay was used to measure cell viability. Western blot was used to measure the protein expression levels of inflammatory indicators (NLRP3, Caspase-1, and ASC), autophagic indicators (LC3B, Beclin-1, and P62), and TLR4.
		                        		
		                        			RESULTS:
		                        			After stimulation with 1 μg/mL LPS for 12 hours, the levels of inflammatory indicators (NLRP3, Caspase-1, and ASC), autophagic indicators (LC3B, Beclin-1, and P62), and TLR4 increased and reached the peak (P<0.05). Compared with the LPS group, the 3-MA+LPS group had reduced expression of autophagy-related proteins and increased expression of inflammation-related proteins and TLR4, while the RAPA+LPS group had increased expression of autophagy-related proteins and reduced inflammation-related proteins and TLR4 (P<0.05). The TLR4 overexpression+LPS group had reduced autophagy-related proteins and increased inflammation-related proteins compared with the TLR4 overexpression control+LPS group, and the TLR4 silencing+LPS group had increased autophagy-related proteins and reduced inflammation-related proteins compared with the TLR4 silencing control+LPS group (P<0.05).
		                        		
		                        			CONCLUSIONS
		                        			In the LPS-induced inflammatory response of human alveolar epithelial A549 cells, autophagic flux has a certain protective effect on A549 cells. TLR4-mediated autophagic flux negatively regulates the LPS-induced inflammatory response of A549 cells.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			A549 Cells
		                        			;
		                        		
		                        			Autophagy
		                        			;
		                        		
		                        			Beclin-1/metabolism*
		                        			;
		                        		
		                        			Caspase 1/metabolism*
		                        			;
		                        		
		                        			Inflammation
		                        			;
		                        		
		                        			Lipopolysaccharides/pharmacology*
		                        			;
		                        		
		                        			NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*
		                        			;
		                        		
		                        			Toll-Like Receptor 4/metabolism*
		                        			
		                        		
		                        	
9.Application of oblique lateral fusion combined with lateral plate fixation in the treatment of adjacent segment diseases of lumbar spine
Yonghui ZHAO ; Sheng LU ; Hui ZHONG ; Tiannan ZOU ; Jie LIU ; Jiayu CHEN ; Zhi PENG
Chinese Journal of Orthopaedics 2022;42(19):1262-1272
		                        		
		                        			
		                        			Objective:To investigate the clinical efficacy of oblique lateral interbody fusion (OLIF) combined with lateral plate (LP) fixation and posterior lumbar interbody fusion (PLIF) combined with pedicle screw fixation in the treatment of adjacent segment disease (ASDis).Methods:Data of 21 ASDis patients treated with OLIF-LP from August 2016 to October 2019 were selected, including 9 males and 12 females; age was 59.3±7.0 years (range, 46-71 years). Target segments: L 2, 3 1 cases (4.8%), L 3, 4 16 cases (76.2%), L 4, 5 4 cases (19.1%). Twenty-one ASDis patients matched with age, sex and surgical segment and treated with PLIF were selected as the control group. The operation time, intraoperative bleeding, postoperative hospital stay, visual analogua scale (VAS), Oswestry disability index (ODI), disc height (DH), intervertebral foramen height (IFH) and lumbar lordosis (LL) were compared between the groups were tested by t-test. VAS score, ODI, DH, IFH and LL were compared within the group by ANOVA, and Bonferroni's test was used for pairwise comparison. Results:All of 42 patients were followed up for 23.7 ±7.4 months (range, 12-36 months). The operation time (97.6 ± 18.0 min) and interpretative bleeding (38.5±62.7 ml) in OLIF-LP group were significantly lower than those in PLIF group (operation time 154.6±42.4 min) and interpretative bleeding (288.6±55.3 ml). There were significant differences between two groups ( t=5.66, P<0.001; t=8.23, P<0.001); the postoperative hospital stay 4.4±1.3 d in OLIF-LP group was longer than that in PLIF group 5.1±1.2 d, but there was no significant difference ( t=1.93, P=0.061); VAS score in OLIF-LP group at 1 month and 3 months after operation (1.6 ± 0.9 points, 1.4 ± 0.8 points), and the ODI index (29.4%±4.7%) after one month operation was improved better than that of PLIF group ( t=2.48, P=0.017; t=2.35, P=0.024; t=2.28, P=0.029), but there was no significant difference between the 12 months after operation of two groups ( t=0.99, P=0.329; t=0.86, P=0.395). The immediately after operation, 3 months after operation and 12 months after operation of DH, IFH and LL in the two groups were significantly improved compared with those before operation ( P<0.05). The immediately after operation, 3 months after operation and 12 months after operation of DH and IFH in the OLIF-LP group were better than those in the PLIF group ( P<0.05), while LL had no significant difference ( P>0.05). There were 2 cases (9.52%) in each group with cage sinking, but no clinical symptoms occurred. In the OLIF-LP group, there was no injury of blood vessels, nerves or abdominal organs during operation, and 2 patients had transient lower limb pain after operation; In the PLIF group, 2 cases (9.52%) of dural rupture were repaired during operation, and no cerebrospinal fluid leakage occurred after operation; Postoperative lower limb pain was aggravated in 3 cases, and improved after dehydration, anti-inflammatory and analgesic treatment; 2 cases of incision exudation healed after symptomatic treatment. Conclusion:OLIF combined with LP fixation has the same clinical effect as PLIF in the treatment of lumbar ASDis, but OLIF combined with LP fixation has more advantages in surgical trauma, postoperative recovery and related complications.
		                        		
		                        		
		                        		
		                        	
10.Analysis of clinical phenotype and genotype of Chinese children with disorders of sex development.
Hu LIN ; Hao YANG ; Jun Fen FU ; Jin Na YUAN ; Ke HUANG ; Wei WU ; Guan Ping DONG ; Hong Juan TIAN ; De Hua WU ; Da Xing TANG ; Ding Wen WU ; Li Ying SUN ; Ya Lei PI ; Li Jun LIU ; Li Ping SHI ; Wei GU ; Lu Gang HUANG ; Yi Hua WANG ; Lin Qi CHEN ; Hong Ying LI ; Yang YU ; Hai Yan WEI ; Xin Ran CHENG ; Xiao Ou SHAN ; Yu LIU ; Xu XU ; Shu LIU ; Xiao Ping LUO ; Yan Feng XIAO ; Yu YANG ; Gui Mei LI ; Mei FENG ; Xiu Qi MA ; Dao Xiang PAN ; Jia Yan TANG ; Rui Min CHEN ; Mireguli MAIMAITI ; De Yun LIU ; Xin Hai CUI ; Zhe SU ; Zhi Qiao DONG ; Li ZOU ; Yan Ling LIU ; Jin WU ; Kun Xia LI ; Yuan LI
Chinese Journal of Pediatrics 2022;60(5):435-441
		                        		
		                        			
		                        			Objective: To explore the heterogeneity and correlation of clinical phenotypes and genotypes in children with disorders of sex development (DSD). Methods: A retrospective study of 1 235 patients with clinically proposed DSD in 36 pediatric medical institutions across the country from January 2017 to May 2021. After capturing 277 DSD-related candidate genes, second-generation sequencing was performed to analyzed the heterogeneity and correlation combined with clinical phenotypes. Results: Among 1 235 children with clinically proposed DSD, 980 were males and 255 were females of social gender at the time of initial diagnosis with the age ranged from 1 day of age to 17.92 years. A total of 443 children with pathogenic variants were detected through molecular genetic studies, with a positive detection rate of 35.9%. The most common clinical phenotypes were micropenis (455 cases), hypospadias (321 cases), and cryptorchidism (172 cases) and common mutations detected were in SRD5A2 gene (80 cases), AR gene (53 cases) and CYP21A2 gene (44 cases). Among them, the SRD5A2 mutation is the most common in children with simple micropenis and simple hypospadias, while the AMH mutation is the most common in children with simple cryptorchidism. Conclusions: The SRD5A2 mutation is the most common genetic variant in Chinese children with DSD, and micropenis, cryptorchidism, and hypospadias are the most common clinical phenotypes. Molecular diagnosis can provide clues about the biological basis of DSD, and can also guide clinicians to perform specific clinical examinations. Target sequence capture probes and next-generation sequencing technology can provide effective and economical genetic diagnosis for children with DSD.
		                        		
		                        		
		                        		
		                        			3-Oxo-5-alpha-Steroid 4-Dehydrogenase/genetics*
		                        			;
		                        		
		                        			Child
		                        			;
		                        		
		                        			China/epidemiology*
		                        			;
		                        		
		                        			Cryptorchidism/genetics*
		                        			;
		                        		
		                        			Disorders of Sex Development/genetics*
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Genital Diseases, Male
		                        			;
		                        		
		                        			Genotype
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Hypospadias/genetics*
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Membrane Proteins/genetics*
		                        			;
		                        		
		                        			Penis/abnormalities*
		                        			;
		                        		
		                        			Phenotype
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Steroid 21-Hydroxylase/genetics*
		                        			
		                        		
		                        	
            
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