1.Study on the expression of IkappaB-alpha protein in TNF-alpha induced apoptosis of U937 cells.
Weihua CHEN ; Guangjie PENG ; Aiping TANG ; Kaiyan WANG ; Muxiang ZHOU ; Lingyi WANG
Chinese Journal of Hematology 2002;23(7):353-355
OBJECTIVETo investigate the TNF-alpha induced apoptosis of U937 cells, the expression, degradation and subcellular localization of IkappaB-alpha, and its degradation mechanism.
METHODChanges and subcellular loca-lization of IkappaB-alpha were observed by fluorescence microscopy, expression and degradation of IkappaB-alpha protein with N-tosyl-L-phenylalanylchloromethyl ketone (TPCK protease inhibitor) blocking test and apoptosis of U937 cell by flow cytometry.
RESULTS(1) immunolfluorescence assay showed that IkappaB-alpha localized in cytoplasm only. (2) The level of IkappaB-alpha protein was downregulated after TNF-alpha stimulation, flow cytometry also confirmed the downregulation. (3) The downregulation of IkappaB-alpha protein levels in TNF-alpha induced apoptosis was partially inhibited by TPCK. (4) The apoptosis rate of U937 cells induced by TNF-alpha was (60.73 +/- 1.61)%.
CONCLUSION(1) Degradation of IkappaB-alpha protein during TNF-alpha induced apoptosis of U937 cells suggested the activation of NF-kappaB. (2) TPCK sensitive protease plays an important role in the degradation of IkappaB-alpha protein. (3) TPCK sensitive protease also involved in the apoptosis of U937 cells induced by TNF-alpha.
Apoptosis ; drug effects ; Down-Regulation ; Humans ; NF-kappa B ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; U937 Cells
2.Correlation of Glut-1, Glut-3 and HK-H expression with 18F-FDG uptake in non-small cell lung cancer and pulmonary inflammatory lesions
Zhenguang WANG ; Mingming YU ; Guangjie YANG ; Yu HAN ; Yangyang WANG ; Peng ZHAO
Chinese Journal of Nuclear Medicine and Molecular Imaging 2018;38(9):605-608
Objective To analyze the expression of glucose transport protein (Glut)-l,Glut-3 and hexokinase (HK)-Ⅱ in non-small-cell lung cancer (NSCLC) lesions and pulmonary inflammatory lesions and discuss the correlation of them with 18F-fluorodeoxyglucose (FDG) uptake.Methods Twenty-four patients with NSCLC and 22 patients with pulmonary inflammatory lesions (25 males,21 females;age range:37-81 years) who underwent PET/CT from November 2012 to May 2016 were retrospectively analyzed.All patients had surgery and were confirmed by pathology.The expression of Glut-1,Glut-3 and HK-Ⅱ in the lesions was detected by immunohistochemistry.Immunohistochemical staining scores and maximum standardized uptake value (SUVmax) were calculated.One-way analysis of variance,the least significant difference t test,two-sample t test and Spearman correlation analysis were used.Results The SUVmax of NSCLC lesions was 8.71 ± 7.62,higher than that of pulmonary inflammatory lesions (3.29 ± 2.16;t =3.220,P< 0.05).Immunohistochemical staining scores of Glut-1,Glut-3 and HK-Ⅱ were 3.75±0.99,4.04±1.00 and 4.00±0.78 for NSCLC lesions respectively,and were all higher than those of pulmonary inflammatory lesions (2.32±0.65,2.89±0.83,2.41±0.50;t values:5.340,5.160,8.130,all P<0.01).The expression of Glut-1 and HK-Ⅱ was positively correlated with SUVmax in NSCLC lesions (rs values:0.414,0.457,both P<0.05).The expression of Glut-1,Glut-3 and HK-Ⅱ was not correlated with SUVmax(rs values:0.392,0.070,-0.066,all P>0.05),but the expression of Glut-3 was higher than that of Glut-1 and HK-Ⅱ (F=4.123,t values:0.970,0.150,all P<0.05) in pulmonary inflammatory lesions.Conclusions The expression of Glut-1,Glut-3 and HK-Ⅱ is higher in NSCLC lesions than that in pulmonary inflammatory lesions.Glut-1 and HK-Ⅱ are the important factors for 18F-FDG uptake in NSCLC.Glut-3 may play an important role in 18F-FDG uptake in pulmonary inflammatory lesions.
3.Application of failure mode and effect analysis in low-energy X-ray radiotherapy
Yining YANG ; Song WANG ; Qingfeng LIU ; Xinyuan GONG ; Mu LI ; Nana LI ; Bo JIANG ; Yuna PENG ; Ping SHEN ; Yu ZHU ; Guangjie YUAN ; Wen SHEN
Chinese Journal of Radiation Oncology 2021;30(3):266-271
Objective:To explore the application of failure mode and effects analysis (FMEA) in low-energy X-ray intraoperative radiotherapy (IORT), analyze its potential risks in IORT, and preliminarily explore the feasibility of FMEA in optimizing IORT management and reducing the occurrence of potential risks.Methods:An FMEA working group was established by the IORT team (1 radiologist, 1 radiology physicist, 2 surgeons, and 2 nurses) to apply the FMEA methodology to conduct a systematic risk assessment. The process modules were established, the potential failure modes and causes for each module were analyzed, the severity (SR), frequency of occurrence (OR) and likelihood of detection (DR) of failure modes were scored and the risk priority number (RPN) was calculated: RPN= SR × OR × DR. The possible errors and potential clinical impact of each part of the radiotherapy process were prospectively analyzed and understood, the causes and current measures were analyzed for each failure mode and preventive measures were proposed and risk management measures were taken accordingly.Results:The IORT process was divided into 8 modules with 14 failure modes. The highest OR value was unsatisfactory target area confirmation (7 points), the highest SR value was equipment failure to discharge the beam (10 points), the highest DR value was wrong key entry after dose calculation (7 points), the highest RPN values were unsatisfactory target area confirmation (210 points) and ineffective protection of endangered organs (180 points). Weaknesses were corrected according to priorities, workflows were optimized and more effective management methods were developed.Conclusion:FMEA is an effective method of IORT management and contributes to reducing the occurrence of potential risks.
4.Cytogenetic Diagnosis of Fanconi's Anemia-Distinguishing Fanconi's Anemia from Aplastic Anemia
Yan XIE ; Hong XIA ; Youhua CHEN ; Guangjie PENG ; Zhiliang XU
Journal of Experimental Hematology 2000;8(3):216-221
Fanconi's anemia (FA) is an autosomal recessive disease featuring a great diversity of clinical symptoms, including congenital malformation, growth retardation and bone marrow failure. Cells obtained from FA patients show a specific hypersensitivity to crosslinking agents such as mitomycin C (MMC). In this study, MMC-induced chromosome breakage tests have been done on 27 healthy controls and 51 patients with bone marrow failure [including 48 patients with aplastic anemia (AA) and 3 patients with FA before cytogenetic analysis]. The results showed that: (1) Diagnosis of 4 FA cases was confirmed, and one of them was the correction of clinical misdiagnosis; bone marrow failure combined with congenital malformation was observed in a few of non-FA aplastic anemia patients, while 1 FA patient lacked congenital abnormality and underdiagnosed before cytogenetic analysis. The data confirmed that misdiagnosis or underdiagnosis of FA could be caused without cytogenetic study. (2) Spontaneous chromosome breakages observed in FA patients were the same as those in AA patients and healthy controls. MMC-induced chromosome breakages observed in FA patient cells were much higher than those in AA patients and healthy controls, especially, metaphases containing more than 5 breakages were easily found in FA lymphocytes treated with 50 ng MMC. (3) Mosaic was found in one of the 4 FA patients. MMC-induced chromosome breakage test at different MMC concentrations could help to dignosis of FA mosaic patient.
5.Clinicopathological features and prognosis of pleomorphic giant cell adenocarcinoma of the prostate
Lang LANG ; Peng HE ; Xiaozhou ZHOU ; Youli WU ; Guangjie DUAN ; Xiaochu YAN ; Zhiwen CHEN
Chinese Journal of Urology 2020;41(7):516-521
Objective:To investigate the clinicopathological features and prognosis of pleomorphic giant cell adenocarcinoma (PGCA) of the prostate, and to strengthen the understanding of this rare variant.Methods:From January 2009 to December 2019, 383 pathological samples of prostate adenocarcinoma with Gleason scores of 8-10 were selected from the First Affiliated Hospital, Army Medical University. PGCA was screened by reviewing the histomorphology of hematoxylin and eosin stained sections. Then the expression of prostate specific markers and mismatch repair (MMR) proteins of PGCA were detected by immunohistochemistry (IHC), and microsatellite instability (MSI) status was detected through polymerase chain reaction (PCR)-capillary electrophoresis. Meanwhile, the clinicopathological characteristics, diagnosis, treatment and prognosis of PGCA were summarized and analyzed along with those reported in the literature.Results:Three patients with PGCA of the prostate were 68, 63 and 71 years old respectively, and case 1 had a history of transurethral resection of the prostate and oral bicalutamide 3 months before surgery. All 3 patients underwent radical prostatectomy and received endocrine therapy, radiotherapy and/or chemotherapy, and died at 18, 23, and 10 months after surgery, respectively. Histologically, both the usual prostate adenocarcinoma with Gleason score of 9-10 and the pleomorphic giant cell component with anaplastic characteristics were observed in 3 tumors, and the latter accounted for 90%, 10%, and 20%, respectively. Immunohistochemical staining showed that both components expressed epithelial markers (CK, CK8/18) and prostate-specific markers (NKX3.1, PSA, P504S) to varying degrees, and the expression of MMR proteins (MSH2, MSH6, MLH1 and PMS2) were not defective. MSI was not detected in the usual prostate adenocarcinoma and pleomorphic giant cell components obtained by microdissection in 3 cases. Combined with 10 cases reported in the literature, there were totally 13 cases of PGCA for reviewing. The patients were 45-81 years old, the average age was 66 years old, and the median age was 66 years old. During the follow-up period of 3-36 months, 7 cases relapsed/metastasized, 6 cases died within 23 months after diagnosis, and 4 of which died within 1 year.Conclusions:PGCA is a newly recognized rare variant of prostate adenocarcinoma. At present, all cases are accompanied with high-grade usual prostate adenocarcinoma with Gleason score of 9-10, but it is different from the latter in pathological morphology and clinical manifestations, by presenting high invasiveness and poor prognosis. PGCA is not sensitive to conventional endocrine therapy, radiotherapy or chemotherapy. Accurate diagnosis of PGCA is helpful to judge the prognosis of patients and guide the treatment.
6.High preoperative neutrophile-lymphocyte ratio predicts poor prognosis of patients undergoing radical cystectomy for nonurothelial carcinoma of the bladder
Shuai LIU ; Xiaozhou ZHOU ; Guangjie DUAN ; Yuan LIU ; Peng HE ; Lang LANG ; Zhiwen CHEN
Journal of Modern Urology 2023;28(11):964-969
【Objective】 To investigate the predictive value of high preoperative neutrophile-lymphocyte ratio (NLR) for the prognosis of nonurothelial carcinoma of the bladder (NUBC) after radical cystectomy (RC). 【Methods】 Clinical and follow-up data of NUBC patients undergoing RC during Jan.2005 and Dec.2020 were collected. The optimal cut-off value of NLR was determined with the receiver operating characteristic (ROC) curve. The survival curve was drawn with Kaplan-Meier method to compare the differences in cancer specific survival (CSS) and overall survival (OS) between the high-NLR and low-NLR groups. The independent risk factors of CSS and OS were screened with Cox proportional hazard regression model. 【Results】 Of the 62 eligible cases,34 (54.8%) were diagnosed with adenocarcinoma,17 (27.4%) with squamous cell carcinoma, 6 (9.7%) with small cell carcinoma and 5 (8.1%) with sarcoma. Kaplan-Meier analysis results showed high NLR was associated with poor CSS (P=0.001) and OS (P<0.001). Cox regression results indicated that high NLR (HR=2.42, 95%CI: 1.12-5.23, P=0.025) and advanced pathologic tumor stage (HR=3.21, 95%CI:1.53-6.74,P=0.002) were independent risk factors of unfavorable CSS. Similarly, high NLR (HR=2.75, 95%CI: 1.35-5.56, P=0.005) and advanced pathologic tumor stage (HR=2.81, 95%CI:1.43-5.57, P=0.003) were independent risk factors of unfavorable OS. 【Conclusion】 As an independent risk factor of unfavorable CSS and OS in NUBC patients undergoing RC, high preoperative NLR is of great value in the prediction of long-term prognosis and may help to optimize individualized treatment.