3.Clinicopathological study on proteomic screening for proteins associated with colorectal carcinogenesis.
Zhi-Hong LIU ; Liang ZENG ; Jian LIU
Chinese Journal of Oncology 2007;29(2):135-136
Adenocarcinoma
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metabolism
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pathology
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Adenoma
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metabolism
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pathology
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Colonic Neoplasms
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metabolism
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pathology
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Cytoplasm
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metabolism
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Female
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HSP27 Heat-Shock Proteins
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Heat-Shock Proteins
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analysis
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Humans
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Intestinal Mucosa
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metabolism
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pathology
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Male
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Neoplasm Proteins
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analysis
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Proteome
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analysis
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Proteomics
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methods
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Rectal Neoplasms
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metabolism
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pathology
4.ADVANCED GLYCATION END PRODUCTS MODIFIED PROTEIN UP-REGULATES EXPRESSION OF ADHESION MOLECULES ON HUMAN ENDOTHELIAL CELLS
Hong LIU ; Fanfan HOU ; Mi LIANG
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
Enhanced expression of adhesion molecules on human endothelial cells of synovial tissue has been demonstrated in patients with dialysis related amyloidosis(DRA).The study was conducted to elucidate the mechanism by which the expression of adhesion molecules on human endothelial cells was up regulated. Human endothelial cells derived from umbilical veins(HUVEC) were coincubated in vitro with native human serum albumin(HSA) or HSA modified with advanced glycation end products(AGE HSA).The expression of adhesion molecule intercellular adhesion molecule 1(ICAM 1),vascular cell adhesion molecule 1(VCAM 1) and E selectin was determined respectively by immunofluorescence staining and flow cytometer analysis.The results showed that ICAM 1 and VCAM 1, but not E selectin,were constitutively expressed on HUVEC. AGE HSA enhanced the expression of ICAM 1, VCAM 1 and E selectin on HUVEC in a time and dose dependent manner. HSA had no effect on the expression of adhesion molecules. The results led to the conclusion that AGE HSA up regulated the expression of adhesion molecules on human endothelial cells and therefore promoted the infiltration of monocytes/macrophages around ? 2 microglobulin amyloid.
6.Effects of T20N Site-directed Mutation on GTPase Activities of OsRacD from Oryza sativa
Xiao-Fei LIU ; Wei-Hong LIANG ;
China Biotechnology 2006;0(01):-
OsRacD belonging to rice Rho family of the small GTP binding proteins,is a pivotal gene involved in rice photoperiod fertility conversion of photoperiod sensitive genic male sterile rice,which influences rice fertility via controlling the pollen tube growth.T20N site-directed mutation was introduced into its highly conserved G1 motif by PCR-mediated method to mimic its GDP-binding state based on the sequences alignment and conserved domains analysis.The prokaryotic expression vector of OsRacD and T20N-OsRacD were constructed,and the His6 tag fused proteins were expressed and purified from E.coli.After identified by Western blot,the GTP hydrolysis activities were detected.The results showed that the GTPase activities of T20N-OsRacD were significantly reduced comparing with that of OsRacD,suggested that OsRacD and T20N-OsRacD have different biochemical characteristics.
7.Effect of selective cycloxygenase-2 inhibitor NS-398 on IL-1α and TNF-α expression in HaCaT cells induced by ultraviolet rays
Yinghua SONG ; Hong LIANG ; Xing DAI ; Xiaoming LIU ; Shahshan WANG
Chinese Journal of Medical Aesthetics and Cosmetology 2012;18(1):52-55
Objective To study the effect of selective COX-2 inhibitor NS-398 on IL-1α and TNF a expression in HaCaT cells induced by UVA/UVB,and further to explore the mechanism on anti human skin photo damage. Methods The subcuhured HaCaT cells were divided into three groups:simple illuminated group was exposed to UVA/UVB directly,NS-398 interfered group was exposed to UVA/UVB after being treated with NS-398 in different concentrations,and the control group was cultured in normal without any treatment.The expression of IL -1α and TNF-α in supernarant was detected by ELISA kit.Results The level of IL-1α and TNF-α expression in supernatant from simple illuminated group was remarkably higher than that in the control group,NS-398 interfered group showed much lower level than the simple illuminated one,and the expression of IL-1α and TNF-α was dependent on the concentration of NS-398.Conclusions NS-398 can reduce IL-1α and TNF-α expression in HaCaT cells induced by ultraviolet rays,suggesting the possibility of anti human skin photo- damage effect.
8.Effect of selective cycloxygenase-2 inhibitor NS-398 on IL-10 and IFN-γ mRNA expression in hunman keratinocytes induced by long-wave ultraviolet
Shanshan WANG ; Hong LIANG ; Xing DAI ; Xiaoming LIU ; Yinhua SHONG
Chinese Journal of Medical Aesthetics and Cosmetology 2012;18(3):204-207
Objective To study the effect of selective COX-2 inhibitor NS398 on IL-10 and IFN-γexpression in human keratinocytes induced by UVA,and further to explore the effect on anti human skin photoaging.Methods HaCaT cells were divided into three groups:simple UV exposure group,NS398-intervene group and blank group.For UV radiation,simple UV exposure group and NS398-intervene group were irradiated with UVA at a dose of 30 J/cm2.In order to assess the effect of NS398,HaCat cells were treated with NS398 at the dose of 0,20,40,80μmol/L respectively and then incubated without irradiation for 2h,substrate changed and cultured for 24h,and then the expression of IL-10 and IFN-γ mRNA was detected via RT-PCR.Results Simple UV exposure group had obviously higher expression of IL-10 mRNA and lower expression of IFN-γ mRNA than that of control group,while NS398-intervened group had significantly lower expression of IL-10 mRNA and higher expression of IFN-γ mRNA than that of simple UV exposure group,and dose-dependency existed.Conclusions NS398 may delay photo-damage of human skin via inhibiting the expression of IL-10 mRNA and up-regulating the expression of IFN-γ mRNA in human keractinocytes.
9.A preliminary proteome analysis on the fluconazole-resistant strains of Penicillium marneffei
Donghua LIU ; Ming LIANG ; Hong LUO ; Shengshun TAN
Chinese Journal of Zoonoses 2010;(1):36-40
The correlated proteome of Penicillium marneffei resistant to fluconazole was investigated in the present study, in which 11 strains of P. marneffei of both the mycelial and yeast forms sensitive to fluconazole were cultivated in Sabouraud's liquid medium containing 8 g fluconazole and the MICs of fluoconazile to P.marneffei before and after inducing cultures were tested by E-test method. The strains of the mycelial and yeast forms showing the most significant increase in MIC value were selected and the protein clip CM10 was used to detect the proteome differences before and after inducing cultures. It was found that 11 strains of the mycelial forms and 2 strains of the yeast forms could tolerate the action of fluconazole and could grow at a drug concentration of 8 μg/mL. Furthermore, the MICs of fluoconazole to the mycelial forms were significantly increased after 7 days of inducing culture and the geometric means of MICs were increased from 1.22 μg /mL to 55.56 μg/mL. After the mycelial forms had been induced to be resistant, 16 proteins were highly expressed with specific expression of the 4581.1 Da and 6109.7 Da proteins; whereas 22 resistant yeast form were highly expressed with specific expression of the 3575.2 Da, 8507.0 Da and 8563.3 Da proteins. These results suggest that the mycelial form of P.narneffei seems to be more tolerant to the action of fluconazole than the yeast form. The resistance of these organisms to fluconazole may be associated to some specific proteins.
10.Effects of serum from myocardial infarction rats on rat bone marrow mesenchymal stem cell differentiation into cardiomyocytes
Jianhua LIU ; Liang HONG ; Xueyun ZHONG ; Jiayu CHEN ; Yunxian CHEN
Chinese Journal of Tissue Engineering Research 2009;13(14):2797-2800
BACKGROUND: Differentiation of bone marrow mesenchymal stem calls(BMSCs) is associated with their microenvironment. After acute myocardial infarction (AMI), the necrosis of cardiomyocytes caused activation of complement, generation of free radical, and secretion of various cell factors. As a result, the ingredients of patients' serum also changed, so does the changes will influence the differentiation of BMSCs into cardiomyocytes? And what influence it will be? OBJECTIVE: To investigative effects of AMI rat serum on the differentiation of BMSCs into cardiomyocytes. DESIGN, TIME AND SETTING: The in vitro controlled cytology experiment was performed at the Laboratory of Center for Stem cells and Tissue Engineering, Sun Yat-sen University from September in 2005 to June in 2006. MATERIALS: SPF Sprague Dawley rats, weighing 50-80 g, aged 3-4 weeks, were used for culture of BMSCs, and SpragueDawley rats, weighing 200-300 g, aged 6-8 weeks, were used for making AMI models. METHODS: BMSCs were isolated and cultured with the adherent method. After making AMI rat models by deligating anterior descending of the left coronary artery, the serum were collected and centrifuged from AMI and normal rats. Then the passage 3 of BMSCs were divided into six groups, non-induction group, 5-azacytidine (5-aza) group, 5-aza plus serum from AMI rat group, 5-aza plus serum from normal rat group, serum from AMI rat group, and serum from normal rat group. MAIN OUTCOME MEASURES: Morphology changes in rat BMSCs after induction; Expression of cardiac troponin T of BMSCs after induction; Expression of GATA-4 and desmin mRNA of BMSCs after induction. RESULTS: After being induced by 5-aza and two kinds of serum, some cells became elongated and thinner. Two to three weeks after induction, some cells had a ball-like or rod-like appearance, and connection were formed between adjacent cells. It showed that some BMSC have differentiated into cardiac like cells. The expression of cardiac troponin T, GATA-4 and desmin were positive in cells differentiated from BMSCs. The troponin T expression in control group and simple serum induction group were negative, but GATA-4 and desmin expressed weakly. CONCLUSION: Serum from AMI rat cannot induce BMSCs to differentiate into cardiomyocytes alone, but it promotes BMSCs induced by 5-aza differentiating into cardiomyocytes and facilitates the differentiated calls into mature.