1.Comparison of two methods of isolation and culture of osteoclasts and the dynamic observation of bone resorption
Binbin LI ; Shifeng YU ; Shuzhen PANG
Journal of Peking University(Health Sciences) 2003;0(05):-
Objective: To study the origin, morphological structure, and functional regulation of osteoclast(OC) for further investigation on the mechanism and regulation of bone resorption. Methods: The OCs were isolated by two kinds of traditional method. Osteoclasts were isoclated from neonatal rat long bones. The cytochemistry was observed. The osteoclast-like cells (OLC) were derived from the mouse bone marrow cells in the presence of 1,25(OH)2VitD3 in vitro. Results: Both morphological and functional studies showed that the isolated cells shared some of the typical characteristics of osteoclasts, that is A. multinuclearity; B. developing spreading and pseudopodial activity when cultured on glass; C. high tartrate-resistant acid phosphatase (TRAP); D. resorption lacunae could be found when the cells were cocultured with devitalized bone slices and the number was increased as the time followed. OLC had the same histological and structural traits as the OCs by the former method. The concentration of Ca 2+ and acid phosphatase (ACP) increased gradually. Conclusion: Different kinds of method fit different experiments. The OC obtained by the first method has more activity of bone resorption. The OLC by the second method has more in quantity and can be used in the study of cell differentiation.
2.Screening of Decolorizing Agents for the Extracts from the Fruit and Stem of Schisandra chinensis
Junxia LIU ; Shifeng PANG ; Fengming DOU ; Yinping JIN ; Yushuai WANG ; Yingping WANG
China Pharmacy 2015;(25):3541-3544
OBJECTIVE:To screen decolorizing agents suitable for the extracts from the fruit and stem of Schisandra chinen-sis. METHODS:HPLC was adopted to determine the contents of 4 kinds of lignans(schizandrin,schisandrol B,deoxyschizandrin and γ-schizandrin) in the extract solution from the fruit and stem of S. chinensis which was treated with 8 kinds of decolorizing agents (activated clay,activated carbon,diatomite,calcium bentonite,kaolin,activated aluminium oxide,magnesium oxide,at-tapulgite clay),and the decolourization rates of the samples of the fruit and stem of S. chinensis and the retention rates of lignans in such samples were calculated respectively. RESULTS:The above-mentioned decolorizing agents were arranged in order as fol-lows respectively based on the decolourization effects on the extract solution from the fruit and stem of S. chinensis:attapulgite clay>activated carbon>activated aluminium oxide>kaolin>magnesium oxide>diatomite>calcium bentonite>activated clay,and activated carbon>diatomite>attapulgite clay>magnesium oxide>kaolin>activated aluminium oxide>activated clay>calcium ben-tonite. The attapulgite clay and activated carbon have the best decolourization effects on the extracts from the fruit and stem of S. chinensis,with the decolourization rates of 60.47% and 69.24% respectively,and the retention rates of schizandrin,schisandrol B,deoxyschizandrin and γ-schizandrin were 77.43%,77.73%,77.07%,77.53%,and 72.18%,70.17%,70.32%,70.28%,re-spectively. CONCLUSIONS:Among the 8 decolorizing agents,attapulgite clay and activated carbon have the best decolourization effects on the extract solution from the fruit and stem of S. chinensis.
3.Relationship of C677T polymorphisms of MTHFR gene with diabetic nephropathy in southeast Guangxi
Kanghai LIU ; Dazhi PANG ; Chaoshi ZOU ; Shifeng WU ; Ping ZHANG ; Qiyan ZENG
International Journal of Laboratory Medicine 2014;(13):1670-1672
Objective To study the relationship between the C677T polymorphisms of N5,N10-methylenetetrahydrofolate re-ductase(MTHFR)gene and type 2 diabetic nephropathy(T2DN).Methods By using PCR-RFLP method,the C677T polymor-phisms of MTHFR was analyzed in 163 cases of type 2 diabetes mellitus(T2DM),which included diabetic nephropathy(DN group, n=82)and diabetes without complications(DM group,n=81),and 77 cases of healthy people as the control group(CON).Plasma total Hcy levels were also measured for all the subjects.Results The frequencies of MTHFR TT and CT homogenetic type and al-lele T(4.9%,37.8%,23.8%)in the DN group were significantly higher than those in the DM(2.5%,28.4%,16.7%)group or the CON group(0.0%,29.8%,14.9%).However,there was no significant difference in MTHFR genotype and allele frequency be-tween the DM group and the CON group.Moreover,plasma Hcy levels were markedly higher in the patients with allele TT and CT genotype than those in the patients with CC genotype,the difference showing statistical significance (P <0.01).The univariate lo-gistic regression analysis showed that the C677T polymorphism of MTHFR gene was closely associated with the development of DN.The odds ratio was 1 .660 and the 95% confidence interval was 1.038 and 2.655 respectively.Conclusion The C677T polymor-phisms of MTHFR gene is associated with T2DN in Southeast Guangxi and the allele T might be the susceptibility gene for DN.
4.In vitro study of the effects of Epimedium on osteoclastic bone resorption in various oral mineralized tissues.
Jingjing LI ; Shifeng YU ; Tiejun LI ; Shuzhen PANG
Chinese Journal of Stomatology 2002;37(5):391-394
OBJECTIVETo investigated the bone resorption caused by osteoclasts and modulating functions of a Chinese herb, Epimedium on bone resorption.
METHODSOsteoclasts were separated from long-limb bones of new born rabbits, cultured with de-activated human tooth slices and glass slices, and treated with different concentrations of Epimedium. Osteoclasts on glass slices were stained by HE staining, TUNEL staining and absorption pits on tooth slices were observed by light microscope.
RESULTSHE staining shows that the Epimedium treated cells showed shrinkage of cytosol and condensation of nuclei. Some treated cells showed breakage of nuclei. TUNEL staining shows that treated cells showed shrinkage of cytosol, dark stained nuclei and light stained cytosol. These results indicate that Epimedium can induce osteoclast to commit apoptosis. Further study shows that the number and area of absorption pits formed on treated tissues are significantly different from those on control tissues. So the effect of Epimedium on bone resorption is dose-dependent.
CONCLUSIONSEpimedium can induce osteoclast apoptosis and inhibit bone resorption. The effect of Epimedium on bone resorption is dose-dependent.
Animals ; Animals, Newborn ; Apoptosis ; drug effects ; genetics ; Bone Resorption ; prevention & control ; Cell Culture Techniques ; methods ; Cells, Cultured ; DNA Fragmentation ; drug effects ; Dose-Response Relationship, Drug ; Epimedium ; In Situ Nick-End Labeling ; Osteoclasts ; cytology ; drug effects ; Plant Extracts ; pharmacology ; Rabbits
5.Effect of light intensity on growth and quality of Asarum heterotropoides var. mandshuricum.
Zhiqing WANG ; Peihe ZHENG ; Shifeng PANG ; Yiming GUAN ; Jing GUO ; Yingping WANG
China Journal of Chinese Materia Medica 2011;36(12):1558-1567
To study the infection rate of leaf spot disease, the drying rate of root and volatile oil content of Asarum heterotropoides var. mandshuricum at the unwrapping stage, blooming stage, the initial fruit stage, fructescence and wither stage during the growth period under the different sunlight intensity of 100% (I), 50% (II), 28% (III), 12% (IV). The volatile oil content was measured according to the method of Chinese Pharmacopoeia and the oil composition was determined by GC-MS. The unwrapping stage, blooming stage and the early fruit stage postponed about 2 days with decrease of the sunlight intensity. The infection rate of leaf was 88.46%, 70.00%, 0.23%, 0.07% under light intensity of I, II, III and IV, respectively, the drying rate was 25.14%, 28.27%, 30.23%, 31.57% under light intensity of I, II, III and IV, respectively, and the volatile oil content was 18.1, 17.6, 16.3, 15.3 mL x kg(-1) under light intensity of I, II, III and IV, respectively. The composition of the oil determined by GC-MS was different between the groups, but the content did not changed significantly with the decrease of the light intensity.
Asarum
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growth & development
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radiation effects
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Light
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Regression Analysis
6.Plant as bioreactor.
Jing YANG ; Tianhang LI ; Lidong XIONG ; Shifeng PANG ; Xiaokun LI
Chinese Journal of Biotechnology 2009;25(5):650-657
Plant can be used as bioreactor for heterogenous protein expression. We reviewed different expression systems of plant bioreactor as well as recent relevant developments. In addition, we discussed perspectives in combination with our own experience.
Bioreactors
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microbiology
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Biotechnology
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trends
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Plants
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genetics
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metabolism
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Plants, Genetically Modified
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metabolism
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Protein Engineering
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methods
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Recombinant Proteins
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biosynthesis
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genetics