1.The expression of costimulatory molecules CD86 in acute myelogenous leukemic cells and clinical implication
Meixia YU ; Yongming ZHOU ; Yuzhen QIU ; Chunhong YAO ; Hao ZHOU
Journal of Chinese Physician 2011;13(4):437-440
Objective To investigate costimulatory molecules CD80 and CD86 expression in acute myelogenous leukemic cells and clinical implication.MethodsThe expression of CD80 and CD86 in the patients with acute myelogenous leukemia and HL-60 cells,U937 cells,NB4 cells,K567 cells was confirmed by Flow Cytometer.ResultsCD80 was very low or no expression in patients with acute myelogenous leukemia.CD86 expressed in acute myelogenous leukemia (27.86±19.65)%,which was much higher than that in control group[(1.21±0.13)%,t=3.55,P<0.01].No significant changes were observed in the expression of CD86 in M4 cells(48.65±21.92)%,M5 cells(39.25±18.67)% and control group(50.20±20.31)%(P>0.05).After the cells were cultured for 24 h and 48 h,the expression of CD86 was (30.62±5.35)% and (29.43±4.67)% in HL-60 cells ,(24.12±5.23)% and (26.56±6.54)% in U937 cells,and,(21.25±3.78)% and (23.21±6.98)% in NB4 cells (all P>0.05).The expression of CD80 and CD86 was very low in K562 cells.ConclusionsCostimulatory molecules CD86 expressed in acute myelogenous leukemic cells in the patients with acute myelogenous leukemia and HL-60 cells,U937 cells,NB4 cells.
2.Protective effects of melatonin on acute cerebral ischemia-reperfusion injury in rats
Yu ZHOU ; Juan YU ; Liying QIU ; Chonghong CHEN
Chinese Pharmacological Bulletin 1987;0(02):-
AIM To study the protective effects of melatonin(MT) on acute cerebral ischemia reperfusion injury.METHODS The model of cerebral ishemia-2 h/reperfusion-24 h was induced by middle cerebral artery occlusion(MCAO) in SD rats. Melatonin (10,20 mg?kg -1 ip)was administered four times in an animal:At 0, 1, 2, 6 h of reperfusion. After 24 h reperfusion, the content of malondialdehyde(MDA),the activities of superoxide dismutase(SOD)and myeloperoxidase (MPO)in brain tissue, the content of thromboxane B 2(TXB 2) and 6-keto-prostaglandin F 1?(6-keto-PGF 1?)in plasma were measured. RESULTS Compared with vehicle group, MT 10, 20 mg?kg -1 protected the activity of SOD, reduced the content of MDA, MT 20 mg?kg -1 also inhibited the increase of MPO in brain tissue ,and attenuated the disequilibrium of TXB 2 /6-keto-PGF 1?.CONCLUSION The protective effects of MT on acute cerebral ischemia reperfusion injury may be related to its increasing antioxidase activities,decreasing lipid peroxidative damage and inhibiting inflammations.
3.Structural feature and biological function of PPP2R5C gene.
Yang-Qiu LI ; Yu-Bing ZHOU ; Li-Jian YANG
Journal of Experimental Hematology 2009;17(5):1127-1129
PPP2R5C is one of the members of regulatory subunits of protein phosphatase 2A (PP2A), which plays a critical role in cell proliferation, differentiation and transformation, based on its induction of dephosphorylation of P53 at various residues. Recently, it was characterized that the alteration of expression pattern of PPP2R5C is associated with cell malignant transformation, thus PPP2R5C was thought as a marker for progressive disease in B-CLL. In this article the gene structure and biological function of PPP2R5C as well as relation of PPP2R5C with genesis and development of cancer were discussed.
Cell Line, Transformed
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Cell Proliferation
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Cell Transformation, Neoplastic
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Humans
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Molecular Structure
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Protein Phosphatase 2
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genetics
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Protein Subunits
4.Down-regulation of midkine gene expression by small interfering RNA affects melanoma cell adhesion and invasion
Yongfing ZHOU ; Dandan GONG ; Zhiyuan QIU ; Huiyong PENG ; Yu FAN
Chinese Journal of Dermatology 2011;44(7):497-500
Objective To study the effects of midkine (MK) gene-targeting small interfering RNA (siRNA)on the invasion of melanoma cells.Methods Three MK gene-targeting siRNAs (S1,S2 and S3)were designed,constructed,and transfected into human A375 melanoma cells.Real-time PCR was performed to measure the expression of MK gene and to screen the siRNA with best efficacy.Then,A375 cells were transfected with the optimal siRNA of various doses (3.125,6.25 and 12.5 nmol/L)followed by additional culture of various durations(24,48,72 hours).Some A375 cells remaining untreated served as the blank control group,and some transfected only with liposomes served as the vector control group.Reverse transcription (RT) -PCR and Western blot were conducted to detect the mRNA and protein expression of MK,respectively,MTT assay to observe the adhesion of A375 cells,and Boyden chamber was used to evaluate cell invasion.Results The expression of MK mRNA was downregulated by all the three siRNAs,especially by the siRNA S3,which was used in the following transfection experiment.Real-time quantitative PCR revealed that the MK mRNA expression was reduced by the siRNA in a dose- (r24hours=-0.906,r4Bhours=-0.922,r72hours=-0.939,all P<0.01)and time-dependent(r3.125nmol/L=-0.889,r625nmol/L=-0.935,r125nmol/L=-0.928,all P<0.01)manner.MTT assay showed that the percentage of adhesing cells was 73.66%±2.25%,49.36%±2.16%and 28.35%±1.68%in A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively.The number of cells migrating across the chamber filter was 23.9±1.6,12.1±1.5,5.6±1.2 among A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively,significantly lower than that in the blank control group(36.8±1.5).The percentage of adhesing cells and number of migrating cells decreased with the dose of siRNA(r=-0.936,-0.915,P<0.01,0.05,respectively).Conclusions MK gene might play an important role in the adhesion and invasion of melanonla cells.To down-regulate the expression of MK gene by siRNA may suppress the adhesion and invasion of melanoma cells.
7.Studies of the expression of CD40~+、CD40L~+ and CD8~+/CD28~+ on peripheral blood cells in patients with chronic B hepatitis
Liann QIU ; Yonglie ZHOU ; Hong YU ; Lijun ZHU ; Hongbo LI
Chinese Journal of Immunology 1985;0(02):-
0.05).There was a positive corrlation on CD40~+ and CD40L~+ in chronic B hepatitis patients.There was a positive corrlation on CD8~+/CD28~+ and CD40~+、CD40L~+ in chronic B hepatitis patients.There were no remarkable corrlation on CD8~+/CD28~- and CD40~+、CD40L~+ in chronic B hepatitis patients.Conclusion:The costimulation molecules CD40~+、CD40L~+ and CD8~+/CD28~+ are lower,while CD8~+/CD28~- are higher in chronic B hepatitis patients than in the health.To test the expression of CD40~+、CD40L~+ and CD8~+/CD28~+ on peripheral blood of the chronic B hepatitis could help to evaluate patients's celluar immunity and guide clinical treatment.
9.Reproductive toxicity of bisphenol A in adult male mice and mechanism
Xingxia HAO ; Dongze ZHANG ; Boyang YU ; Yan ZHANG ; Yuanyuan ZHOU ; Xueying QIU ; Haole ZHOU ; Taodi LIU
Journal of Jilin University(Medicine Edition) 2016;42(2):195-199
Objective:To investigate the damage effect of bisphenol A (BPA)on the testis tissue of adult mice, and to reveal the reproductive toxicity of BPA in the body of animal and mechanism.Methods:40 KM mice aged 8 weeks were randomly divided into control group (according to the weight ratio of corn oil gavage), low dose of BPA group (100 mg·kg-1BPA),moderate dose of BPA group (200 mg·kg-1 BPA),and high dose of BPA group (400 mg·kg-1 BPA).4 weeks laster,the testis tissue was taken.The apoptotic rates in the testis tissue were detected by flow cytometry;the distribution and expression of Fas and FADD were measured by immunohistochemistry.Results:Compared with control group,the apoptotic rate,the expression rates of Fas and FADD in testis tissue of the mice in low dose of BAP group had no changes (P>0.05),while the apoptotic rates in the testis tissue and the positive expressions rates of Fas and FADD in moderate and high doses of BPA groups were increased (P<0.05).Compared with low dose of BPA group,the apoptotic rates and the positive expression rates of FAS and FADD in tests tissue of the mice in moderate and high doses of BPA groups were significantly increased (P<0.05).Compared with moderate dose of BPA group,the apoptotic rate and the positive expression rates of FAS and FADD in testis tissue of the mice in high dose of BPA group was significantly increased (P<0.05).The overexpression of Fas and FADD was positively correlated to the apoptotic rate in testis tissue in moderate dose of BPA group (r=0.430,P<0.05;r=0.238,P<0.01)and high dose of BPA group (r=0.637,P<0.01;r=0.359,P<0.01).Conclusion:BPA with content dose can increase the apoptotic rates of cells in testis tissue and the expressions of Fas and FADD.BPA’s reproductive toxicity may be closely associated with the activation of Fas signal pathway and resulting in massive apoptosis.
10.Influence of TROP-2 gene on the ability of human breast cancer cell metastasis
Li YU ; Yu FAN ; Zhiyuan QIU ; Yongjing ZHOU ; Dandan GONG ; Xiudi XIAO ; Zhengyan WU
Chinese Journal of Primary Medicine and Pharmacy 2011;18(15):2017-2019
ObjectiveTo study the effects of tumor-associated calcium signal transducer-2 (TROP-2) gene small interfering RNA(siRNA) on adhesion and invasion of human breast cancer cell. MethodsReal time PCR was used to evaluate the TROP-2 mRNA of seven human breast cancer cell lines Bcap-37 ,LCC1 ,MCF-7 ,MDA-MB-231,MDA-MB-435, MDA-MB-468 ,and ZR75-1. The cell line of TROP-2 highest expression was transfected with different dose of TROP-2 siRNA. The expression of TROP-2 mRNA and protein were determined by Real-time quantitative PCR and immumoflurescence method. The cell adhesion was evaluated by MTT assay,and invasion was exmined by hoyden chamber,respectively. Results Cell line MCF-7 showed the highest elevation of TROP-2 mRNA in seven breast cancer cell lines. The results from real-time quantitative PCR and immumoflurescence method showed that TROP-2 mRNA and protein reduced in time-and dose-dependent manners( P < 0.01 ;P < 0.01 ). The adhesive rate of siRNA groups(5 nM,10 nM,and 20 nM)was(52.9 +2.5)% ,(25.6 ±2.3)%, ( 12.8 +2.2)% (P <0.01 ) ,respectively.The transwell results showed that the invasion cells was(78 ± 17), (39 ± 15), ( 19 ± 16), ( 136 +25 ) and( 139 ±21 )in different groups(5,10,20 nM siRNA,and controls) ,respectively(P <0.01). ConclusionTROP-2 gene might play an important role in adhesion and invasion of human breast cancer cell. siRNA targeted TROP-2 could effectively inhibit adhesion and invasion of human breast cancer cell.