1.Effects of recombinant hirudin variant Ⅲ on expression of apoptosis-related genes during galactose-mediated human lens epithelial cells damage
Yu, OU ; Zhi-jun, YUAN ; Pei, GENG ; Wu-tong, WU
Chinese Journal of Experimental Ophthalmology 2011;29(7):581-584
Background Recombinant hirudin variant Ⅲ(rHV3) can effectively prevent galactose-induced human lens epithelial cells LECs injury,but little is known about the molecular mechanism of its action.Objective The present study was to investigate the effects of rHV3 on the expression of apoptosis-related genes in damaged LECs induced by galactose.Methods The rHV3 was extracted by our research group,and the biological activity of rHV3 was identified by titration of thrombase according to Markwardt's method.Human LECs (SRA01/04) were cultured using 125×10-3 mol/L D-galactose+10% FBS+D/F12 medium to establish the damaged human LECs model.rHV3 was added into the medium of the damaged human LECs model.Human LECs were cultured in D/F12 medium containing 10% FBS as normal control.The expression of apoptosis-related genes,such as aldose reductase (AR),bax,bcl2 and p53,in LECs at the mRNA level was detected using RT-PCR.The abundance ratio of target genes was presented with the absorbance (A) of gene mRNA/GAPDH mRNA.Results Compared to the normal control group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were significantly elevated in model group (t=3.90E-06,t=8.44E-04,t=5.15E-08,P<0.01).However,in the rHV3-treated group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were lower than those of model group (t=5.90E-06,t=1.51E-04,t=3.42E-06,P<0.01).The bcl2 mRNA/GAPDH mRNA was markedly downregulated in the model group when compared with the normal control group (t=1.86E-05,P<0.01);while after rHV3 addition,bcl2 mRNA/GAPDH mRNA increased in comparison with the model group (t=8.56E-05,P<0.01).Conclusion 125×10-3mol/L D-galactose induces the damage and apoptosis of human LECs.rHV3 likely plays a protective function on D-galactose-induced damage of human LECs by inhibiting the polyol pathway and mitochondria-mediated pathway.
2.Apoptosis or necrosis, should which be expected for tumor cells?.
Ke-Fu WU ; Xiao-Tong MA ; Yu-Hua SONG
Journal of Experimental Hematology 2005;13(6):921-923
Evidence has indicated that low doses of anti-tumor regimens can induce cell apoptosis in vitro, although different regimens induce apoptosis by different mechanism and pathway. In recent years, new tumor treatment strategy has been mainly focused on inducing tumor cell apoptosis. The present review discusses the advantages and disadvantages of inducing tumor cell apoptosis. The benefit of inducing apoptosis is not to cause inflammatory reaction, but as its disadvantage, it inhibits immune responses, and the phagocytosis of apopotic bodies may result in horizontal transfer of genes (including oncogenes and other oncogenic materials), which can be one of the causes of tumor relapse. This paper proposes that the tumor treatment strategy should be turn into promoting tumor cell necrosis and inducing anti-tumor immune responses.
Antineoplastic Agents
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therapeutic use
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Apoptosis
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drug effects
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Humans
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Necrosis
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chemically induced
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Neoplasms
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drug therapy
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immunology
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pathology
3.Gastric collision tumor (adenocarcinoma and gastrointestinal stromal tumor): report of a case.
Jian WU ; Yu-jie HE ; Shu-lan TONG
Chinese Journal of Pathology 2010;39(8):563-564
Adenocarcinoma
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metabolism
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pathology
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surgery
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Aged
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Female
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Gastrectomy
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methods
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Gastrointestinal Stromal Tumors
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metabolism
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pathology
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surgery
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Humans
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Keratins
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metabolism
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Neoplasms, Multiple Primary
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metabolism
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pathology
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surgery
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Proto-Oncogene Proteins c-kit
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metabolism
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Stomach Neoplasms
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metabolism
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pathology
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surgery
7.The fusion and division during osteoclasts formation
Suojing YU ; Peijian TONG ; Chengliang WU ; Hongting JIN ; Letian SHAN
Chinese Journal of Orthopaedics 2013;(1):82-88
Objective To investigate the formation and special cell biological behavior of osteoclasts.Methods The live-cell imaging technology was adopted to consecutively and dynamically observe the whole process of multinuclear osteoclast formation induced by RANKL and M-CSF from rat peripheral blood monocyte.Meanwhile,the inverted phase contrast microscopy,TRAP staining,and scanning electron microscopy were also applied to identify the osteoclast.Results After 2-week cultivation,a great number of apocytes were found by the inverted phase contrast microscopy,and most apocytes and monocytes had positive reaction after TRAP staining.Moreover,many bone resorption lacunae in which osteoclasts were perhrming bone resorption function could be found in the bone slice under the scanning electron microscope.Live-cell imaging observation showed that the multinuclear osteoclasts were generated through self-fusion of monocytes,fusion of monocytes and apocytes,as well as fusion between apocytes.All fusion processes occurred under the condition of cell adherence.Time-lapse Microcinematography observation showed diverse shapes of osteoclasts and the cell division of multinuclear osteoclasts.Conclusion Rat peripheral blood monocyte can develop into osteoclast under induction of RANKL and M-CSF.Osteoclast can form gigantic apocyte via various types of cell fusion to increase its nucleus number and cell volume,vary its shape,and increase the area of plasma membrane.On the other hand,osteoclast can decrease its cell volume and nucleus number via cell division to adapt the needs of local morphology,biomechanics and bone resorption dynamics.It suggests that this non-mitosis cell division is a special cell biological behavior of osteoclast,which may be the basis of exerting its function and improving bone resorption efficiency.
8.Experimental Study of the Effect of Wuyaojing Granules on the Immunological Function of Mice
Guanhai DAI ; Renzhao WU ; Feng YANG ; Yeling TONG ; Yu CHEN
Journal of Medical Research 2006;0(01):-
Objective To observe the effect of Wuyaojing granules(WYJG) on immunological function of mice.Methods Mice were randomly divided into six groups:the high-dosage WYJG group(6.0g/kg),the midst-dosage WYJG group (2.0g/kg),the low-dosage WYJG group(1.0g/kg),the Lindera aggregate group (0.9g/kg),the korean Ginseng group (2.0g/kg) and the normal control group(H2O).After a period of 30-day treatment,the effects of WYJG on the T lymphocyte transformations capacity induced by ConA,delayed hypersensitivity leaded by DNFB,the number of lymphocyte B,serum level of hemolysin and NK activity were observed. Results The high-dosage and the low-dosage WYJG could significantly increase T lymphocyte transformations capacity induced by ConA(P0.05).Conclusion WYJG can significantly increase T lymphocyte transformations capacity induced by ConA and strengthen delay hypersensitivity leaded by DNFB.
9.Study of adenovirus mediated human p27kip1 antioncogene on the apoptosis in esophageal carcinoma cell lines
Qingming WU ; Qiang TONG ; Jieping YU ; Al ET ;
Chinese Journal of Digestion 2001;0(03):-
Objective To study the effects of p27kip1 gene on the apoptosis in esophageal carcinoma cell lines EC 9706. Methods A replication deficient adenovirus vector encoding p27kip1(ad p27kip1) was constructed and human esophageal carcinoma cell lines EC 9706 were transduced in vitro. The cell apoptosis were determined by flow cytometry, TUNEL technique and DNA fragmentation analysis. Results Ad p27kip1 containing the p27kip1 sequence was successfully constructed and the virus titer was 1.24?10 12 CFU/ml. The transduction efficiency was 100% when multplicity of infection≥50. FCM analysis revealed a sub G 1 cell peak in ad p27kip1 transduced esophageal carcinoma cell lines. Agarose electrophoresis showed marked ladder. The difference of apoptotic index between the ad p27kip1 group and the control group was statistically significant(37.3?3.4 vs. 1.3?0.2, P
10.Effecting of inhibiting ubiquitin-proteasome pathway on proliferation and apoptosis of gastric carcinoma cells
Qingming WU ; Jieping YU ; Weiguo ZHANG ; Xianjun LIU ; Qiang TONG ;
Chinese Journal of Digestion 2001;0(02):-
Objective To investigate the effects of inhibiting ubiquitin proteasome pathway(UPP) on proliferation of gastric carcinoma cells and the possible mechanism was discussed. Methods The gastric carcinoma cell strain SGC 7901 was treated with MG 132 to inhibit its UPP specially. The effect of growth suppression on cells was evaluated with MTT assay. Cell cycle and apoptosis were detected by flow cytometry(FCM). DNA fragment analysis was used for confirming the presence of apoptosis. The activity of telomerase was examined by TRAP PCR ELISA. Expression of p27kip1 was detected by immunocytochemical technique. Results MG 132 had great inhibitory effect on the growth of SGC 7901 cells. The FCM analysis showed that the ratio of G0/G1 phase of control group was (46.3?4.1)%, the ratio of G0/G1 phase of SGC 7901 cells treated with MG 132 increased to (72.1?5.0)% ( P