1.Effect of echinacoside-containing serum in promoting mesenchymal stem cell osteogenic differentiation and ZHX₃ expression in rats.
Yuan TIAN ; Yang DI ; Cui-fen BAO ; Yu-han LIN ; Shu-jian QIN
China Journal of Chinese Materia Medica 2015;40(20):4052-4057
To investigate the effect and possible mechanism of echinacoside-containing serum on the osteogenic differentiation in rat bone marrow mesenchymal stem cells. Rat bone marrow mesenchymal stem cells were cultivated by the whole bone marrow adherence method. The 3rd generation of cells were divided into 3 groups: the blank control group, the classic osteogenic-induced group and the 10% echinacoside-containing serum group. The expression of alkaline phosphatase and osteocalcin were detected by ELISA. The ex- pression of ZHX, protein was detected by Western blot technique. RT-PCR technique was used to detect the expression of ZHX₃mRNA. According to the result, the expressions of the alkaline phosphatase and osteocalcin in the classic osteogenic-induced group and the 10% echinacoside-containing serum group were significantly higher than that of the blank control group (P <0. 01). And expressions of the alkaline phosphatase activity and osteocalcin in the 10% echinacoside-containing serum group were significantly higher than that in the classic osteogenic-induced group (P < 0.01). Meanwhile, the classic osteogenic-induced group and the 10% echinacoside-containing serum group showed obviously higher ZHX₃ protain and mRNA expression than that of the black control group, with significant differences (P < 0.01); the 10% echinacoside-containing serum group showed obviously higher ZHX₃ protain and mRNA expression than that of the classic osteogenic-induced group, with a significant difference (P < 0.01). In conclusion, 10% echinacoside-containing serum can promote the differentiation of the bone marrow mesenchymal stem cells cultured in vitro. Its mechanism may be correlated with the increase in the ZHX₃expression.
Animals
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Cell Differentiation
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drug effects
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Cell Proliferation
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drug effects
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Cells, Cultured
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Female
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Glycosides
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blood
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pharmacology
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Homeodomain Proteins
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genetics
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metabolism
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Male
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Mesenchymal Stromal Cells
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cytology
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drug effects
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metabolism
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Osteogenesis
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drug effects
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Rats
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Rats, Sprague-Dawley
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Serum
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chemistry
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Transcription Factors
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genetics
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metabolism
3.Effect of puerarin in myocardial protection in rats with acute and chronic alcoholism.
Journal of Southern Medical University 2011;31(12):2035-2038
OBJECTIVETo investigate the protective effect of puerarin on the myocardium of rats with acute and chronic alcoholism.
METHODSIn acute alcoholism experiment, normal male SD rats were randomly divided into the control group, alcoholism group and puerarin group (n=8), and high- and low-dose puerarin was administered. In chronic alcoholism experiment, increasing puerarin doses were given. Serum and myocardial levels of spartate aminotransferase (AST) and creatine phosphokinase (CPK) were determined using enzymatic methed, and superoxide dismutase (SOD), malondialdehyde (MDA), Ca(2+)-Mg(2+)-ATPase, and Na(+)-K(+)-ATPase in the myocardium were assayed with colorimetric method. HE staining was used to observe the microscopic changes of the myocardium.
RESULTSCompared with alcoholism group, puerarin-treated groups showed significantly lowered myocardial contents of MDA, CPK and AST and serum levels of AST and CPK (P<0.05, P<0.01) and increased myocardial SOD (P<0.05, P<0.01), Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase activity (P<0.05, P<0.01), but Na(+)-K(+)-ATPase was similar between the two groups (P>0.05). HE staining of the myocardium showed cell swelling and obscure cell boundaries in alcoholism group, especially in chronic alcoholism group. The myocardial structure in puerarin group remained clear and regular.
CONCLUSIONPuerarin can protect from myocardial injuries induced by acute and chronic alcoholism in rats.
Alcoholism ; drug therapy ; Animals ; Cardiomyopathy, Alcoholic ; metabolism ; pathology ; prevention & control ; Ethanol ; toxicity ; Isoflavones ; pharmacology ; therapeutic use ; Male ; Rats ; Rats, Sprague-Dawley
4.Comparion of the expression of CS3 fimbriae in different vector systems
Rong-Kai, GAO ; Zhao-shan, ZHANG ; Shu-Qin, LI ; Cui-Fen, HUANG
Bulletin of The Academy of Military Medical Sciences 2001;25(1):1-4
Objective:To choose the best vector for the expression of CS3 fimbriae. Methods: The CS3 operon was cloned into different plasmid vectors such as pUC19 and pTrc99A. The expression of CS3 was monitored by whole-cell ELISA and SDS-PAGE analysis. The assembly of CS3 fimbriae was detected with electron microscopy. Results:The expression level of CS3 fimbriae using plasmid pUC19 as carrying vector was the highest, and the insertion orientation of CS3 gene into the plasmid has a little effect on its expression level. The expression of CS3 fimbriae was confirmed by SDS-PAGE analysis and electron microscopy.Conclusions:The promotor of CS3 itself played the key role in the expression of CS3 fimbriae and the copy number of plasmid was the main factor to affect the expression level.
5. Protective Mechanism of Puerarin on Spatial Memory in Hippocampus of Chronic Alcoholism Rats
Chinese Pharmaceutical Journal 2014;49(18):1610-1614
OBJECTIVE: To investigate the protective mechanism of puerarin on spatial memory in hippocampus of rats suffering from chronic alcoholism.
6.Effects of pravastatin on the expression of endothelin induced by aldosterone in rat cardiac fibroblasts.
Yu-Zhou WU ; Wei CUI ; Shu-Qin LI ; Lei ZHANG ; Jing-Chao LU
Chinese Journal of Applied Physiology 2007;23(3):343-346
AIMTo investigate the effects of pravastatin on endothelin(ET) expression induced by aldosterone in cultured neonatal rat cardiac fibroblasts.
METHODSET concentration in conditioned medium was measured by radioimmunoassay, intracellular ET-1 level was evaluated by flow cytometry, and the expression of preproendothelin-1 (ppET-1) was detected and quantified using reverse transcriptase-polymerase chain reaction (RT-PCR) method.
RESULTSThe cardiac fibroblasts, treated with aldosterone at 107 mol/L, significantly up-regulated ppET-1 mRNA expression, as well as ET-1 synthesis and release. Pravastatin (10(-5), 10(-4), 10(-3) mol/L) dose-dependently blocked these effects. In contrast, pravastatin-induced inhibitory effects were reversed in the presence of mevalonate.
CONCLUSIONPravastatin down-regulated ppET-1 mRNA expression, as well as ET-1 synthesis and release induced by aldosterone in a process specifically related to mevalonate in cardiac fibroblasts.
Aldosterone ; metabolism ; Animals ; Cells, Cultured ; Endothelins ; metabolism ; Fibroblasts ; drug effects ; metabolism ; Myoblasts, Cardiac ; drug effects ; metabolism ; Pravastatin ; pharmacology ; Rats ; Rats, Sprague-Dawley
7.Isolation and structure identification of flavonol glycoside from Lysimachia christinae Hance.
Dong-bin CUI ; Shu-qin WANG ; Ming-ming YAN
Acta Pharmaceutica Sinica 2003;38(3):196-198
AIMTo isolate and prepare chemical standards.
METHODSFlavonol glycoside and rhamnonic acid gamma-lactone were isolated from whole plants of Lysimachia christinae Hance. with 95% ethanol and purified by silica gel column chromatography. UV, IR, MS, 1HNMR, 13CNMR, DEPT, HMQC, HMBC were used for the structural identification.
RESULTSTwo compounds were obtained. They were identified as rhamnonic acid gamma-lactone (I) and kaempferol-3-O-alpha-L-rhamnosyl (1-->2)-beta-D-xyloside (II).
CONCLUSIONCompound I was isolated from this genus for the first time. Compound II is a new compound named as lysimachiin.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Lactones ; chemistry ; isolation & purification ; Molecular Structure ; Plants, Medicinal ; chemistry ; Primulaceae ; chemistry
8.Prokaryotic expression and purification of N-terminal and C-terminal fragments of histone deacetylase 4.
Yang YANG ; Xiao-cui QIN ; Shu-hu LIU ; Wei HUANG ; Xue-min WANG
Journal of Southern Medical University 2010;30(4):712-715
OBJECTIVETo express and purify the fusion proteins of glutathione S-transferase (GST)-N-terminal of histone deacetylase4 (HDAC4-N') (1-1952 bp) and GST- C-terminal of HDAC4 (HDAC4-C') (1708-3255 bp) in E.coli.
METHODSThe DNA fragments (HDAC4-N' and HDAC4-C') amplified by PCR were ligated into GST fusion vector (pGEX-6P-1) to construct the recombinant plasmids. After identification with restriction digestion and DNA sequencing, the recombinant plasmids were transformed into E.coli BL21 and induced by IPTG for their expression. After identification by SDS-PAGE and Western blotting, the target proteins were purified by glutathione sepharose 4B.
RESULTSThe results of restriction digestion and DNA sequencing confirmed successful construction of the recombinant plasmids. The relative molecular masses of the fusion proteins were approximately 110500 and 93080 as shown by SDS-PAGE. Western blotting demonstrated that the fusion proteins could be recognized by the specific anti-HDAC4 antibody.
CONCLUSIONWe have successfully constructed the recombinant expression vectors of pGEX-6P-1/HDAC4-N' and pGEX-6P-1/HDAC4-C' and induced the expression of the fusion proteins, which may facilitate functional studies of HDAC4 with other proteins.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Glutathione Transferase ; biosynthesis ; genetics ; Histone Deacetylases ; biosynthesis ; genetics ; Humans ; Peptide Fragments ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Repressor Proteins ; biosynthesis ; genetics
9.Relationship between intravascular ultrasound imaging features of coronary plaques and soluble CD105 level in patients with coronary heart disease.
Song CUI ; Shu-zheng LÜ ; Yun-dai CHEN ; Guo-xiang HE ; Jian-ping LIU ; Zhi-yuan SONG ; Mao-qin SHU ; Hou-yuan HU ; Bo-li RAN ; Tao JING
Chinese Medical Journal 2007;120(7):595-597
10.Split hand/foot malformation: report of a family with 20 cases.
Anli SHU ; Chua-nan YI ; Miao-miao LIU ; Cui-qin HUANG ; She CHEN ; Shu-mei YANG ; Qiang HE ; Xi-dan LI
Chinese Journal of Medical Genetics 2013;30(4):498-499
Adult
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Female
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Foot Deformities, Congenital
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diagnosis
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genetics
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Hand Deformities, Congenital
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diagnosis
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genetics
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Humans
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Male
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Pedigree
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Young Adult