1.Combined use of RGD-peptide modified PLGA and TGF-beta1 gene transfected MSCs to improve cell biobehaviors in vitro.
Changwen, LI ; Qixin, ZHENG ; Xiaodong, GUO ; Daping, QUAN ; Jie, ZHAO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(5):592-8
In order to improve the surface properties of PLGA polymer for a better material/cell interface to modulate the cells behaviors, we prepared a novel three-block copolymer, PLGA-[ASP-PEG], and immobilized an RGD-containing peptide, Gly-Arg-Gly-Asp-Ser-Pro-Cys (GRGDSPC) on the surface of it. Transforming growth factor-beta1 (TGF-beta1) was transfected into bone marrow stromal cells (MSCs) employed as seeded cells. Cell adhesion, spreading, proliferation and differentiation on this material were investigated. The results showed that the cell adhesive ratio on RGD-modified materials was higher than on un-modified materials (P<0.05). The extent of cell spreading was also wider on RGD-modified materials than on un-modified materials. Cell proliferation indices of transfected MSCs were increased as compared with the un-transfected MSCs (P<0.05). The ALP activities in the MSCs cultured with RGD-modified materials were higher than on un-modified materials after 14 days (P<0.05), and those in transfected MSCs were higher than in un-transfected MSCs (P<0.05). It was suggested that the combined use of RGD-modification and TGF-beta gene transfection could improve the interaction of biomaterial and cells.
2.Acetylization of histone regulated by valproic acid sodium on the regulatory effect of cell cycle related factor
Changwen SHI ; Xia ZHAO ; Lili CA ; Jingjie SUN ; Jie LI
Chinese Journal of Microbiology and Immunology 2008;28(8):760-765
Objective To investigate the regulation on cell cycle related factor such as Cyclins and P21waf/cip1 by inhibiting histone deacetylase(HDAC)with valproic acid sodium(VPA).Methods HepG2 hep-tocellular carcinoma cells.BGC-823 gastric carcinoma cells and MCF-7 breast cancer cells were cultured with O.75-4.00 mmoL/L VPA for 48 h in vivo.and the cell cycle was analyzed by flow cytometry with PI assay.The protein and mRNA expression of Cyclin A,Cyclin D1,Cyclin E and P21waf/cip1 were analyzed by indirect immu-nofluorescence technique and RT-PCR.respectively.Results Compared with control groups,VPA at concen-trations 0.75-4.00 mmol/L exerted a significant inhibiting effect on G1 phase of HepG2,BGC-823 and MCF-7 cells(P<0.001).and the effect was dose dependent.Cyclin A was down-regulated both at mRNA and protein level in HepG2 and BGC-823 cells(P<0.001),but no difference in MCF-7 cells(P>0.05).Cyclin D1 was down-regulated both at mRNA and protein level(P<0.001)and P2lwaf/cip1 was up-regulated both at the mRNA and protein level in the three cell lines(P<0.001);Conversely,protein and mRNA expression of Cyclin E were unchanged upon treatment with VPA(P>0.05).Condusion Acetylization of histone intervened with VPA can regulate Cyclin D1 and P21waf/cip1 expressions obviously.To the expression of Cyclin A,it shows some difference according to the histogenesis and phenotypes of different carcinoma types.But there is not any obvious function on Cyclin E.Down-regulating Cychn D1 and up-regulating P21waf/cip1 may be the common target path-way in the inhibition of cell cycle G1 phase exerted by VPA.
3.THE VISCOSITY MEASUREMENTS OF MOLECULAR WEIGHTS OF POMELO PECTIN
Changwen ZHAO ; Lisha ZHANG ; Jinming LIN ; Tingyuan LI
Academic Journal of Second Military Medical University 1982;0(02):-
The number average molecular weights of five fractionated samples of pomelo pectin were determined osmometrically in aqueous solution. The values of Mn ranged from 4.74?104 to 1.83?144 for different fractions.From the data of osmotic pressure and intrinsic viscosity, the [?]-M relation obtained is[?]3.23?10-7M1.75in 0.9% NaCl solution of pH 4.83 at 37℃.
4.Study of regulating histone acetylizad level with VPA on the proliferation of breast cancer cells
Changwen SHI ; Xia ZHAO ; Jingjie SUN ; Lili CAO ; Zhenhai YU ; He GU
Journal of Chinese Physician 2008;10(7):907-910
Objective To investigate the effect and the mechanism of up-regulating histone acetylizad level with a selective inhibitor of HDACs-Valproate acid sodium (VPA) on breast cancer cell proliferation. Methods MCF-7 cells were cultured with 0.75-4.0 mmol/L valproic acid (VPA) for 24, 48, 72, 96 hours in vitro, the inhibiting rate was tested by MTT assay. Cell cycle was analyzed by flow eytome- try with PI assay, and the protein and mRNA expressions of Cyelin A, Cyclin DI, Cyclin E, P21Waf/cipl of MCF-7 cells after 1.5, 3.0 mmol/ L VPA treated were analyzed by indirect immunofluorescence technique and RT-PCR respectively. Results After cultured with 0.75 -4.0 mmol/L valproic acid (VPA) for 24, 48, 72, 96 hours, the inhibiting rate of experimental groups increased significantly(P<0.01) and a dose and acting time dependent manner was found. As to cell cycle, the percentages of GI, S, M phrase in control groups remained the same. Contrary to control groups, 0. 75 -4.0 mmo]/L VPA induced a significant arrest in G1 phrase ( P<0.01), and a total of 55.4% -82.8% G1 phrase ratio were found. P21Waf/cipl was up-regulated both at the mRNA and protein level while Cyclin D1 was down-regulated ( P<0.001). Conversely, neither mRNA nor protein expression of Cyclin A, Cyclin E showed difference ( P>0.05). Conclusions Up- regulating histone acetylizad level can inhibit breast cancer cell proliferation, induce cell cycle arrest in G1 phrase. VPA, as a I class of histone deaeetylase inhibitor, can be used as an option in the treatment of breast cancer. The mechanism may include up-regulating P21Waf/cipl mRNA and protein expression and down-regulating Cyclin D1 mRNA and protein expression.
5.Analysis of physiological and biochemical characteristics of SPF Yorkshire and Landrace swine
Jinqiang QUAN ; Caixia GAO ; Xinjie JIANG ; Changwen LI ; Xiaoye LU ; Shengguo ZHAO ; Hongyan CHEN
Acta Laboratorium Animalis Scientia Sinica 2017;25(2):160-165
Objective To determine the reproductive physiology and blood physiological and biochemical characteristics of SPF Yorkshire and Landrace swine.Methods Ten reproductive physiology parameters,19 blood physiological parameters and 18 blood biochemical parameters in SPF Yorkshire and Landrace swine were measured using conventional methods and the differences between population,between age groups and between both sexes were analyzed.Results There were no significant differences(P>0.05) in reproductive physiology parameters and most blood physiological and biochemical parameters of the SPF Yorkshire and Landrace swine.A few of parameters,such as blood physiological indices GRAN,HGB,RDW,PLT,PCT,and blood biochemical indices ALKP,CHOL,TBIL,BUN,showed significant difference(P<0.05) between populations,between age groups and between both sexes,however,the values of difference were rather small,deviated from the normal range.Conclusion The physiological and biochemical characteristics of SPF Yorkshire and Landrace swine are basically stable and there is no significant difference compared with other laboratory miniature pigs.This study will provide valuable basic data for raising velvet yield,establishment of animal models and evaluating the genetic quality of closed colony.
6.Modulation of Caspase expression by up-regulating histone acetylizad level with VPA
Xia ZHAO ; Changwen SHI ; Jingjie SUN ; Lili CAO ; Jie LI ; Zhenhai YU
Chinese Journal of Immunology 1985;0(03):-
Objective:To investigate modulation of a specific HDAC inhibitor,Valproate acid sodium(VPA),on expression of Caspase3,Caspase8,Caspase9 by inhiting HDAC,as well as apoptosis rate of cancer cells treated with VPA and the specific inhibitors of Caspase3,Caspase8,Caspase9.Methods:Heptocellular carcinoma cells-HepG2,gastric carcinoma cells-BGC-823 and breast cancer cells-MCF-7 were cultured with 0.75-4.0 mmol/L of VPA for 48 hrs in vivo,apoptosis was analyzed by flow cytometry with Annexin V/PI assay.The activities and protein expressions of Caspase3,Caspase8,Caspase9 were detected by spectrophotometry and indirect immunofluorescence technique.Results:Contrary to control groups,VPA at concentrations between 0.75 and 4.0 mmol/L induced a significant apoptosis in HepG2,BGC-823,MCF-7 cells(P0.05).The apoptosis rates of cancer cells treated with VPA and specific inhibitors of Caspase3,Caspase9 together was lower than in the groups with VPA treatment singlely(P
7.Dedicated distribution of nerve growth factor in different forms
Jun LIU ; Tianrun Lü ; Xiaojian CAO ; Yongjun LI ; Jianhua LI ; Changwen FU ; Min MIN ; Qi CHEN ; Yi ZHAO ; Leshen YAO
Chinese Journal of Tissue Engineering Research 2005;9(13):166-167
BACKGROUND: That the nerve growth factor(NGF) is capable of treating peripheral nerve injury has been broadly acknowledged. But it is not sure whether it is able to pass through blood brain barrier(BBB) to act on central nerve system. In this study, the NGF encapsulated in liposome was compared with NGF alone in their abilities of passing through BBB.OBJECTIVE: To compare the dedicated distribution of NGF in different forms using single photon emission computerized tomography(SPECT).DESIGN: It is a randomized controlled study with New Zealand rabbits as subject.SETTING: An affiliated hospital of Nanjing Medical College.MATERIALS:The trial was conducted in Nanjing Senke Medical Company and the Nuclear Medicine Department of the First Hospital of Nanjing Medical College from June 2003 to May 2004. The subjects were 19 New Zealand rabbits of either sex, weighting (2.0 ±0. 2) kg, from Anlimo Technology Company. [99Tcm] -NGF(labeling rate 98.9%, purity 99.7% )was made by Senke Medical Company. Liposome was provided by Shengyang Pharmaceutical College. Urethine solution(200 g/L) was from Pharmacy Department of NanJing Medical Collage.METHODS: [99Tcm]-NGF was encapsulated in liposome and was treated as the following: The liposomesA containing 1.48 × 108Bq[99Tcm] -NGF were injected into rabbits and its distribution percentage was analyzed with SPECT. The same amounts of[99Tcm] -NGF and[99Tcm] -NGF-ordinary liposomesB were treated in the same way.MAIN OUTCOME MEASURES: The ratio of concentration and radiation percentage of NGF in brain to those in the whole body.RESULTS: The[99Tcm]-NGF encapsulated in self-made liposomeA presented high radiation in the brain. But[99Tcm] -NGF alone was almost completely excreted through urinary system and[99Tcm] -NGF encapsuled in ordinary liposomeB was mostly phagocytized by liver reticuloendothelial system.CONCLUSION: The self-made NGF-liposomeA is brain-dedicated, which set a basis for drugs to pass through BBB.
8.Sequencing and Phylogenetic Analyses of Structural and Accessory Proteins of Middle East Respiratory Syndrome Coronavirus from the First Imported Case in China, 2015.
Roujian LU ; Lirong ZOU ; Yanqun WANG ; Yanie ZHAO ; Weimin ZHOU ; Jie WU ; Wenling WANG ; Guizhen WU ; Changwen KE ; Wenjie TAN
Chinese Journal of Virology 2015;31(4):333-340
To investigate the genetic character and origin of the first imported infection case of middle East respiratory syndrome coronavirus (named as MERS-CoV_China GD01), RNA was extracted from swabs of this patient followed by RT-PCR amplification. All coding gene of structural (S, E, M, E) and accessory (ORF3, ORF4a, ORF4b, ORF5, ORF8b) proteins were sequenced and analyzed. Phylogenetic analyses of structural protein coding genes of MERS-CoV_ China GD01 indicates that several substitutes exists in S coding gene and its origin belong group 5 of MERS-CoV, which were recent circulated in Saudi Arabia area, while other three structural genes (N, E, M) were very conserved. Phylogenetic analyses of accessory protein coding genes of MERS-CoV China GD01 indicates that several substitutes exists among ORF3, ORF4a, ORF4b and ORF5, while ORF8b was conserved. In conclusion, genome of MERS-CoV_ China GD01 was general conserved although several genetic variations were found among structural and accessory protein coding genes. This is the first report on sequencing and phylogenetic analyses of the first imported MERS case in China, which may pay the way for prevention and control of imported MERS-CoV infection.
China
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Conserved Sequence
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Coronavirus Infections
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transmission
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virology
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Evolution, Molecular
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Genomics
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Humans
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Middle East Respiratory Syndrome Coronavirus
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genetics
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physiology
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Phylogeny
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Sequence Analysis
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Viral Proteins
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genetics
9.Combined Use of RGD-peptide Modified PLGA and TGF-β1 Gene Transfected MSCs to Improve Cell Biobehaviors in vitro
LI CHANGWEN ; ZHENG QIXIN ; GUO XIAODONG ; QUAN DAPING ; ZHAO JIE
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(5):592-598
In order to improve the surface properties of PLGA polymer for a better material/cell in-terface to modulate the cells behaviors, we prepared a novel three-block copolymer, PLGA-[ASP-PEG],and immobilized an RGD-containing peptide, Gly-Arg-Gly-Asp-Ser-Pro-Cys (GRGDSPC) on the sur-face of it. Transforming growth factor-131 (TGF-β1) was transfected into bone marrow stromal cells (MSCs) employed as seeded cells. Cell adhesion, spreading, proliferation and differentiation on this material were investigated. The results showed that the cell adhesive ratio on RGD-modified materials was higher than on un-modified materials (P<0.05). The extent of cell spreading was also wider on RGD-modified materials than on un-modified materials. Cell proliferation indices of transfected MSCs were increased as compared with the un-transfected MSCs (P<0.05). The ALP activities in the MSCs cultured with RGD-modified materials were higher than on un-modified materials after 14 days (P<0.05), and those in transfected MSCs were higher than in un-transfected MSCs (P<0.05). It was suggested that the combined use of RGD-modification and TGF-β gene transfection could improve the interaction of biomaterial and cells.