2.The effects of IGF-1 on the proliferation and apoptosis of cultured condylar chondrocytes
Rui LUO ; Ruizhi ZHANG ; Zhenglin GONG ; Ping ZHANG
Journal of Practical Stomatology 2016;32(6):810-814
Objective:To investigate the effects of IGF-1 and IL-1β on the proliferation and apoptosis of cultured human condylar chondrocytes( CCs) of temporomandibular joint( TMJ) . Methods:Cultured CCs were derived from human TMJ condylar cartilage tis-sue, and identified by immunocytochemistry staining. The cultured cells were divided into 6 groups:control group, IL-1β(10 μg/L) group and IL-1 group (10 μg/L) + IGF-1 group (0, 1, 10, 50 and 100 μg/L, respectively). The cell proliferation ability was de-tected by MTT assay. The cell cycle and apoptosis were detected by flow cytometry. The expression of apoptosis-associated factors Bcl-2, Bax and p38 MAPK/NF-κB proteins were detected by Western blot. Results:Type II collagen was positively expressed in cultured CCs. IL-1β treatment decreased cell proliferation, increased cell apoptosis with concomitant increase of the percentage of early apopto-sis and late apoptotic cells, increased Caspase-3 expression, decreased Bcl-2/Bax ratio, and increased the expression of p38 MAPK/NF-κB proteins. Whereas, with 1-100 μg/L IGF-1 pretreatment, the proliferation ability and Bcl-2/Bax ratio of the cells were in-creased(P<0. 05), the apoptotic cells were decreased, the expression of Caspase-3 and p38 MAPK/NF-κB proteins was decreased ( P<0. 05) in a dose-dependent manner. Conclusion:IGF-1 may inhibit IL-1β-induced cell apoptosis and attenuate the activation of p38 MAPK/NF-κB of human condylar chondrocytes.
3.The effects of siRNA targeting Mcl-1 on biological behavior of salivary adenoid cystic carcinoma SACC-2 cells
Ruizhi ZHANG ; Ping ZHANG ; Bo YU ; Rui LUO ; Zhenglin GONG
Journal of Practical Stomatology 2014;(6):809-812
Objective:To explore the effect of siRNA targeting myeloid cell leukemia-1(Mcl-1)on the biological behavior of salivary adenoid cystic carcinoma cells.Methods:The chemically synthesized Mcl-1-siRNA was transfected into salivary adenoid cystic carci-noma SACC-2 cells.The expression levels of Mcl-1-mRNA and Mcl-1protein were examined by Real-time PCR and western blotting respectively.MTT assay,transwell chamber and flow cytometry were used to determine the effect of Mcl-1-siRNA on SACC-2 cell pro-liferation,migration and apoptosis.Results:Compared with the control group,liposome group and NC-siRNA group,SACC-2 cell proliferation rate of Mcl-1-siRNA group was obviously slowed down.48 h after transfection,the migration of SACC-2 cells in Mcl-1-siRNA group(39 ±9.0)were lower than that in control group(69 ±6.0).The apoptosis rate of Mcl-1-siRNA group(8.6%)was sig-nificantly higher than that in control group(1.9%).Conclusion:Silence Mcl-1 can inhibit cell proliferation and migration and pro-mote apoptosis of salivary adenoid cystic carcinoma cells.
4.c-fos modulates p16/CyclinD1 signaling pathways and promotes the proliferation and migration of oral squamous cell carcinoma
Rui LUO ; Ruizhi ZHANG ; Zhenglin GONG ; Ping ZHANG
Journal of Practical Stomatology 2017;33(1):100-104
Objective:To investigate the effect of c-fos on the proliferation and migration of oral squamous cell carcinoma and potential mechansism.Methods:The expression of c-fos,CyclinD1 and p16 in 60 oral squamous cell carcinoma samples and 60 oral mucosa tissue samples was examined by immunohistochemistry.HN6 and SCC9 cells were respectively transfected with siRNA-c-fos and siRNA-scramble,then were respectively divided into control group,siRNA-scramble group and siRNA-c-fos group.The mRNA and protein expressions of CyclinD1 and p16 were decteted,meanwhile cell proliferation and migration were tested.Results:Compared with the oral mucosa tissue samples,the expressions of CyclinD1 and c-fos were increased in the carcinoma samples,while the expression of p16 was reduced.Compared with control group,the expressions of CyclinD1 in siRNA-c-fos group were significantly reduced,while p16 enpression was increased,with the inhibition of cell proliferation and migration.Conclusion:c-fos may regulate pl6/CyclinD1 signaling pathways and promote the proliferation and migration of oral squamous cell carcinoma.
6.The effects of Cyr6 1 on the biological behavior of human adenoid cystic cancer cells
Ruizhi ZHANG ; Ping ZHANG ; Bo YU ; Rui LUO ; Zhenglin GONG
Journal of Practical Stomatology 2014;(4):526-529
Objective:To explore the effects of cysteine-rich 6 1 (Cyr6 1 )on biological behavior of human adenoid cystic carcinoma ACC-LM and ACC2 cells.Methods:The chemically synthesized Cyr6 1-siRNA was transfected into ACC-LM and ACC2 cells.Cell proliferation was measured by the MTT method,the invasive ability was evaluated by Transwell chamber assay,and cell apoptosis was analyzed using flow cytometry by double staining with Annexin V and propidium iodide.Results:Cyr61-siRNA significantly down-regu-lated Cyr61 protein expression in ACC-LMand ACC2 cells.Cyr61-siRNA markedly inhibited the proliferation and invasion of the cells, however,there was no significant difference in cell apoptosis between Cyr6 1-siRNA and control groups.Conclusion:Cyr6 1 promote the proliferation and invasion of adenoid cystic cancer cells.
7.The correlation between point mutation of MBL ExonI and its plasma concentration in North Huis
Ping LI ; Tianjun JIA ; Wanyuan SHI ; Rui HAN ; Qiang LUO ; Shumin ZHANG
Chinese Journal of Immunology 2001;0(07):-
Objective:The major aim of this study is to analyze the point mutation at the codon 54th of MBL in healthy North Huis,and to measure the plasma levels of MBL, and to analyze the association between the mutation frequency and plasma MBL concentrations.Methods:PCR-RFLP was used to detect MBL point mutation.MBL plasma concentrations were measured using MBL Oliger ELISA kit.Results:Frequency of point mutation at the codon 54th of MBL in healthy Huis was 0.15. The plasma MBL concentration was (3.40?2.55)mg/L. There was negative correlation between MBL concentrations and gene mutation frequency in huis(r=-0.67).Conclusion:The relationship between frequency of mutation at codon 54 of MBL gene and the plasma MBL concentrations in healthy Huis is negative correlation.
8.Characterization and genomic analysis of bacteriophage SH-Kp152234 infecting extensively drug-resistant Klebsiella pneumonia
Ye ZHI ; Tingting LUO ; Rui WANG ; Xijia FENG ; Ke LIU ; Chenkun SHI ; Lulu WANG ; Fupin HU ; Ping HE
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(3):273-277
Objective · To identify broad-spectrum bacteriophages against extensively drug-resistant Klebsiella pneumonia and analyze their characteristics by biological and genomic methods. Methods · Multi-drug resistant Klebsiella pneumonia strains collected from a hospital were used as host bacteria to isolate and purify broad-spectrum phages in the wastewater at the same hospital area. The size and shape of phages were observed by transmission electron microscope. Titer, host range, pH stability and thermal stability were measured. Moreover, the DNA extracted from the phage SH-Kp152234 was sequenced and analyzed. Results · One strain of bacteriophage against Klebsiella pneumonia was isolated and named as SH-Kp152234. The electron microscope revealed it belongs to Podoviridae family. Moreover, genome of SH-Kp152234 showed to be a linear double-stranded DNA of 40578 bp with the GC content of 52.85%. It was predicted to have 49 open reading frames with related known functions.Conclusion · SH-Kp152234, with a broad host range and a short latent period, which could exert its activity in a wide range of temperature and pH, is a promising candidate to be exploited in the treatment of multiple drug-resistant Klebsiella pneumonia.