1.Contribution' of autophagy inhibitor to radiation sensitization in nasopharyngeal carcinoma cells
Zhirui ZHOU ; Xiaodong ZHU ; Wei ZHAO ; Song QU ; Wenyan PAN ; Ya GUO ; Fang SU ; Xiaoyu LI
Chinese Journal of Radiological Medicine and Protection 2012;32(5):449-454
Objective To investigate the role of autophagy in radiation-induced death response of human nasopharyngeal carcinoma cells.Methods MTT method was used to detect cell viability of CNE-2 cells in different time after irradiation.Clonogenic survival assay was used to evaluate the effect of autophagy inhibitor (chloroquine phosphate) and autophagy inductor (rapamycin) on radiosensitivity of nasopharyngeal carcinoma cells.Cell apoptosis was assessed by flow cytometry.The expressions of LC3 and P62 were measured with Western blot.Cell ultrastructural analysis was performed under an electron microscope.Results Irradiation with 10 Gy induced a massive accumulation of autophagosomes accompanied with up-regulation of LC3-Ⅱ expression in CNE-2 cells.Compared with radiation alone,chloroquine phosphate (CDP) enhanced radiosensitivity significantly by decreasing cell viability (F =25.88,P < 0.05),autophagic ratio (F =105.15,P < 0.05),and LC3-Ⅱ protein level(F =231.68,P <0.05),while up-regulating the expression of P62 (F =117.52,P < 0.05).Inhibition of autophagy increased radiation-induced apoptosis (F =143.72,P < 0.05).Rapamycin (RAPA) also significantly decreased cell viability,but increased autophagic ratio and LC3-Ⅱ protein level while down-regulated the expression of P62.Induction of autophagy increased radiation-induced apoptosis(F =167.32,P < 0.05).Conclusions Blockage of autophagy with CDP could enhance radiosensitivity in human nasopharyngeal carcinoma cells,suggesting that inhibition of autophagy could be used as an adjuvant treatment to nasopharyngeal carcinoma.
2.Effect of BMP9 over-expression on migration and invasion of human lung squamous-cell carcinoma NCI-H520 cells
Jing WANG ; Fang DENG ; Ya LI ; Mengtian FAN ; Yangliu GUO ; Qiong SHI
Chinese Journal of Pathophysiology 2015;(11):1921-1927
AIM:To investigate the effect of bone morphogenetic proteins 9 (BMP9) on the migration and in-vasion abilities of human lung squamous-cell carcinoma NCI-H520 cells and its mechanism.METHODS:The expression of BMP9 at mRNA and protein levels in the NCI-H520 cells and human bronchial epithelial ( HBE) cells was detected by RT-PCR and Western blot.The NCI-H520 cells were transfected with the recombinant adenovirus AdBMP9 and the expres-sion of BMP9 at mRNA and protein levels was validated by RT-PCR and Western blot.The migration and invasion abilities of the NCI-H520 cells were determined by wound-healing and Transwell assays.The mRNA and protein levels of the migra-tion-related factor matrix metalloproteinase 2 (MMP2) were detected by RT-PCR and Western blot.The level of phospho-rylated Smad1/5 (p-Smad1/5) was detected by Western blot.Meanwhile, NCI-H520 cells were treated with BMP specific antagonist AdNoggin and AdBMP9.The level of p-Smad1/5 and the cell migration ability were measured by Western blot, wound-healing and Transwell assays.RESULTS:The expression of BMP9 at mRNA and protein levels was lower in NCI-H520 cells than that in HBE cells.After AdBMP9 was stably transfected into the NCI-H520 cells, the expression of BMP9 at mRNA and protein levels was significantly up-regulated, cell migration and invasion abilities were significantly de-creased, and the mRNA and protein levels of MMP2 were decreased.Meanwhile, the level of p-Smad1/5 was increased. Noggin reversed BMP9-caused the increase in p-Smad1/5 and the decrease in cell migration ability.CONCLUSION:O-ver-expression of BMP9 inhibits the migration and invasion abilities of lung squamous-cell carcinoma NCI-H520 cells.The activation of BMP-Smad signaling pathway may be involved in this inhibitory process.
3.Comparative proteomic analysis on radioresistant nasopharyngeal carcinoma cell
Xiaodong ZHU ; Shiting HUANG ; Song QU ; Fang SU ; Ya GUO ; Jinzi WANG
Chinese Journal of Radiological Medicine and Protection 2012;32(3):245-248
Objective To discover radioresistance-associated proteins by performing comparative proteomic analysis on nasopharyngeal carcinoma cell lines.Methods The total proteins were extracted from radioresistant human nasopharyngeal carcinoma cell line CNE-2R and its parental cell line CNE-2,respectively.These proteins were separated by high quality two-dimensional polyacrylamide gel electrophoresis (2-DE) and then the 2-DE profiles were screened for differentially expressed protein spots by the Image Master 5.0 software.Those spots were identified by a matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry.Results 32 significantly differentially expressed protein spots were screened in two different radiosensitivity cell lines and 11 proteins were identified by tandem mass spectrometry,among which 3 proteins were up-regulated in radioresistant human nasopharyngcal carcinoma cell line CNE-2R and the other 8 proteins were down-regulated.Conclusions The differentially expressed proteins of nasopharyegeal carcinoma cells with different radiosensitivity were mainly involved in apoptosis regulation,DNA damage and repair,cell cycle regulation,RNA transcription,cell signaling,cytoskeleton formation and radiation stress responses.
4.Radioresistance-related signaling pathways in nasopharyngeal carcinoma cells
Ya GUO ; Xiaodong ZHU ; Song QU ; Fang SU ; Qi WANG ; Wei ZHANG
Chinese Journal of Radiological Medicine and Protection 2011;31(2):167-171
Objective To study the difference of gene expression profile between the radioresistant human nasopharyngeal carcinoma cell line CNE-2R and CNE-2,and to screen the signaling pathway associated with radioresistance of nasopharyngeal carcinoma.Methods The radioresistant nasopharyngeal carcinoma cell line CNE-2R was constructed from the original cell line CNE-2.CNE-2R and CNE-2 cells were cultured and administered with 60Co γ-ray irradiation at the dose of 400 cGy for 15 times.Human-6v 3.0 whole genome expression profile was used to screen the differentially expressed genes.Bioinformatic analysis was used to identify the pathways related to radioresistance.Results The number of the differentially expressed genes that were found in these 2 experiments was 374.The Kegg pathway and Biocarta pathway analysis of the differentially expressed genes showed the biological importance of Toll-like receptor signaling pathway and IL-1 R-mediated signal transduction pathway to the radioresistance of the CNE-2R cells and the significant differences of 13 genes in these 2 pathways,including JUN,MYD88,CCL5,CXCL10,STAT1,LY96,FOS,CCL3,IL-6,IL-8,IL-1α,IL-1B,and IRAK2(t=13.47-66.57,P<0.05).Conclusions Toll-like receptor signaling pathway and IL-1R-mediated signal transduction pathway might be related to the occurrence of radioresistance.
5.Experimental study of antibiotics and anticoagulants effection on mouse peritonaeum
Yi-Ling ZHANG ; Jian-Wu TANG ; Zhen-Bo DONG ; Ya-Fang GUO ;
Chinese Journal of Clinical Pharmacology and Therapeutics 1999;0(04):-
Aim The effects of antibiotics and anticoagulants on mouse peritonaeum were ob-served to explore the factor of the peritoneal dialysis related sclerosing peritoni-tis. Methods The experimental models of peritoneal dialysis were established in miceby infusing different kind of drugs to the peritoneal cavity and the changes of the peri-toneal membrane for each drug at different time were observed by the autopsy and lightmicroscope for several weeks. Results Amikacin, Cefradine, Zinacef, Ciprofloxacin,Heparin and Urokinase could induce sclerosing changes of peritoneal membrane such asloss of peritoneal mesothelum infiltration of inflammatory cells and of proliferation fibrecell.These changes were irreversible after the drugs were stoped.Conclusion Thedrugs commonly used in peritoneal dialysis may in different degree result in peritonealsclerosis.
6.Experience for Education Course of Aids Among Medical College Students
Li-Jun XIAO ; Yan-Hui FANG ; Ya-Chun GUO ; Li-Xin LIU ;
Microbiology 1992;0(06):-
Developing AIDS education course in college and university is urgent and important,especially for medical college students on account of the characteristic of their specialty.This paper is on the basis of practice and consideration for AIDS education course among medical college students.
7.Tumor mass in left chest wall.
Ren-ya ZHANG ; Jing GUO ; Xi-chao SUN ; Fang-fang XU ; Hong PAN ; Chuan-tao YUAN ; Peng ZHU
Chinese Journal of Pathology 2008;37(2):139-141
8.Cellular fatty acids as chemical markers for differentiation of Acinetobacter baumannii and Acinetobacter calcoaceticus.
Chao YANG ; Zhao Biao GUO ; Zong Min DU ; Hui Ying YANG ; Yu Jing BI ; Gui Qin WANG ; Ya Fang TAN
Biomedical and Environmental Sciences 2012;25(6):711-717
OBJECTIVEGas chromatography (GC) was used to investigate the cellular fatty acid (CFA) composition of 141 Acinetobacter baumannii and 32 A. calcoaceticus isolates from different locations in China and to find chemical markers to differentiate these two closely related bacteria.
METHODSWhole cell fatty acid methyl esters (FAMEs) were obtained by saponification, methylation, and extraction for GC analysis, followed by a standardized Microbial Identification System (MIS) analysis.
RESULTSAll A. baumannii and A. calcoaceticus strains contained some major fatty acids, namely, 18:1 ω9c, 16:0, Sum In Feature 3, 12:0, 17:1ω8c, 3-OH-12:0, 17:0, Sum In Feature 2, 2-OH-12:0, and 18:0 compounds. Although most of the total CFAs are similar between A. baumannii and A. calcoaceticus strains, the ratios of two pairs of CFAs, i.e., Sum In Feature 3/18:1 ω9c versus 16:0/18:1 ω9c and Sum In Feature 3/18:1 ω9c versus unknown 12.484/18:1 ω9c fatty acids, could differentiate these two closely related bacteria. A. baumannii could be easily classified into two subgroups by plotting some ratios such as Sum In Feature 3/16:0 versus 17:0 and Sum In Feature 3/2-OH-12:0 versus 17:0 fatty acids.
CONCLUSIONThe ratios of some CFAs could be used as chemical markers to distinguish A. baumannii from A. calcoaceticus.
Acinetobacter baumannii ; classification ; cytology ; metabolism ; Acinetobacter calcoaceticus ; classification ; cytology ; metabolism ; Biomarkers ; metabolism ; Fatty Acids ; metabolism ; Species Specificity
9.Double mulching application for Panax notoginseng growing seedlings.
Xiao-Hong OU ; Yan FANG ; Ya-Na SHI ; Lan-Ping GUO ; Li WANG ; Yan YANG ; Hang JIN ; Da-Hui LIU
China Journal of Chinese Materia Medica 2014;39(4):566-571
OBJECTIVEIn order to improve the irrigation for Panax notginseng growing seedlings, different mulching ways were carried out to investigate the effects of double mulching.
METHODField experiment was applied to study soil moisture, soil temperature and bulk density of different mulching ways while the germination rate and seedlings growth also were investigated.
RESULTCompared with the traditional single mulching with pine leaves or straw, double mulching using plastic film combined with pine leaves or straw could reduce 2/3 volumes of irrigation at the early seedling time Double mulching treatments didn't need to irrigate for 40 days from seeding to germination, and kept soil moisture and temperature steady at whole seedling time about 30% and 9.0-16.6 degrees C, respectively. The steady soil moisture and temperature benefited to resist late spring cold and germinate earlier while kept germination regularly, higher rate and seedlings quality. In contrast, single mulching using pine leaves or straw had poor soil moisture and temperature preserving, needed to irrigate every 12-day, meanwhile dropped the germination and booming time 14 days and 24-26 days, respectively, reduced germination rate about 11.3%-8.7%. However, single pine leaves mulching was better than straw mulching. In addition, though better effects of soil moisture and temperature preserving as well as earlier and higher rate of germination with single plastic films mulching had, some disadvantages had also been observed, such as daily soil temperature changed greatly, seedling bed soil hardened easily, more moss and weeds resulted difficulty in later management.
CONCLUSIONTo the purpose of saving water and labor as well as getting higher germination rate and seedlings quality, double mulching using plastic films combined pine leaves at the early time and single mulching removing plastic films at the later time is suggested to apply in the growing seedlings of P. notoginseng.
Agriculture ; methods ; China ; Drugs, Chinese Herbal ; analysis ; Fertilizers ; analysis ; Panax notoginseng ; chemistry ; growth & development ; Quality Control ; Seedlings ; chemistry ; growth & development ; Soil ; chemistry
10.Expression and Significance of Caspase-3 and Survivin at Different Stages of Hemangioma in Children
hong, LIU ; ya-mei, DANG ; ying-fang, GUO ; jin-sui, WANG ; shu-hua, LIAN ; jiang, QIAN ; cheng-gang, QIAO
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To investigate the expression and significance of Caspase-3 and Survivin at different stages of human hemangioma in children.Methods Fifty-five specimens of hemangioma tissue excised in operation and 8 normal skins removed in operation were harvested from People's Hospital of Gansu Province between Jan.2000 and Dec.2005.The pathologic diagnosis was divided into 3 groups according to Mulliken's standard under microscope:proliferated phase group(n=31),degenerated phase group(n=24),and control group(n=8).All these specimens were examined by immunohisto for Caspase-3 and Survivin expression.Results Caspase-3 positive rate in the proliferative phase and involuting phase were 35.5% and 79.2%,respectively.The positive rate in involuting phase was higher than that that in proliferative phase,the difference between the 2 groups was significant(P=0.045 9).A significant difference was not found between the proliferative phase and normal skin tissue(P=0.057 3).Survivin positive rate in the proliferating and involuting phase were 77.4% and 45.8%,respectively.The positive rate in proliferative phase was higher than that in involuting phase,the difference between the 2 groups was significant(P=0.008 5),the difference in involuting phase and normal skin tissue was not significant(P=0.059 3).Conclusions High level of Caspase-3 expression in vascular endothelial cells in involuting phase in contrast to that in proliferative phase,which indicates that Caspase-3 may play a positive role in the apoptosis of vascular endothelial cells.Survivin may inhibit the apoptosis of vascular endothelial cells in proliferative phase due to it's high expression.Caspase-3 and Surivivin involved in the hemangioma in the regulation of apoptosis.J Appl Clin Pediatr,2009,24(1):51-52