1.Chemical constituents from Vaccinium bracteatum.
Jing QU ; Xia CHEN ; Chang-Shan NIU ; Shi-Shan YU
China Journal of Chinese Materia Medica 2014;39(4):684-688
The chemical constituents of Vaccinium bracteatum were studied by means of macroporous resin, ODS column chromatography and preparative HPLC. Eleven compounds were isolated from this plant. By using ESI-MS and NMR, the structures of the eleven compounds were determined as 10-O-trans-p-coumaroyl-6alpha-hydroxyl-dihydromonotropein (1), 10-O-cis-p-coumaroyl -6alpha-hydroxyl-dihydromonotropein (2), vaccinoside (3), 10-O-cis-p-coumaroyl monotropein (4), isolariciresinol-9-O-beta-D-xyloside (5), tectoridin (6), vicenin-3 (7), quercetin-3-O-alpha-L-rhamnoside (8), quercetin-3-O-alpha-L-arabinopyranoside (9), quercetin-3-O-beta-D-galactopyranoside (10), and quercetin-3-O-beta-D-glucuronide (11), respectively. Compounds 1 and 2 are new, and compounds 4, 6 and 7 are isolated from the genus Vaccinium for the first time.
Drugs, Chinese Herbal
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chemistry
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Molecular Structure
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Spectrometry, Mass, Electrospray Ionization
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Vaccinium
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chemistry
2.Effect of aqueous extract of Radix et Rhizome Rhodiolae on expressions of HIF-1?, HIF-1? and VEGF in endothelial cells exposed to hypoxia
Xiufang GAO ; Haiming SHI ; Ying SHAN ; Yong YU ; Xuanchun WANG
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(04):-
Objective: To investigate the effect of aqueous extract of Radix et Rhizome Rhodiolae on expressions of HIF-1?, HIF-1? and VEGF in human umbilical vein endothelial cells(HUVECs) exposed to hypoxia. Methods: The effect of different treatment time and concentrations of aqueous extract of Radix et Rhizome Rhodiolae on cellular proliferation were determined with XTT colorimetric assay. The mRNA and protein expressions of HIF-1?, HIF-1? and VEGF were analyzed respectively by quantitative real-time polymerase chain reaction with SYBR Green I and Western blot. Three groups were studied: normoxia control group, hypoxia control group, Radix et Rhizome Rhodiolae treatment group. Results: The optimal condition of Radix et Rhizome Rhodiolae treatment were 24h and 10?g/mL. The protein levels of HIF-1?, HIF-1? and VEGF were elevated by hypoxia (P
3.Study on secondary metabolites of endophytic fungi Penicillium polonicum.
Jing LIU ; Guang-Zhi DING ; Lei FANG ; Shi-Shan YU
China Journal of Chinese Materia Medica 2014;39(20):3974-3977
The PDB culture medium was selected to ferment the endophyte strain, and the secondary metabolites of endophytic fungi Penicillium polonicum were studied. Combined application of Sephadex LH-20, ODS and HPLC chromatographies over the ethyl acetate extract of the fermented culture led to the isolation of 6 compounds. By spectral methods, the structures were elucidated as [3, 5-dihydroxy-2-(7-hydroxy-octanoyl)]-ethylphenylacetate (1), (3, 5-dihydroxy-2- octanoyl)-ethyl phenylacetate (2), (5, 7-di- hydroxy-9-heptyl)-isobenzo pyran-3-one (3), 3-(hydroxymethyl) 4-(1E)-1- propen-1-yl-(1R, 2S, 5R, 6S)-7-oxabicyclo [4.1.0] hept-3-ene-2, 5-diol (4), (E)-2-methoxy-3-(prop-1-enyl) phenol (5) and p-hydroxylphenylethanol (6).
Biological Factors
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chemistry
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metabolism
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Endophytes
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chemistry
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isolation & purification
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metabolism
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Fabaceae
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microbiology
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Fermentation
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Penicillium
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chemistry
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isolation & purification
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metabolism
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Secondary Metabolism
4.Determination of human telomerase reverse transcriptase and telomerase activity in different tumor cell lines and its significance
Xiaoxia YU ; Yingai SHI ; He DONG ; Lihong ZHANG ; Shan WU
Journal of Jilin University(Medicine Edition) 2006;0(04):-
Objective To detect the expression of humar telomerase reverse transcriptase (hTERT) and telomerase activity in HeLa,MCF-7,SMMC7721 PC-3m and U2OS cell lines. Methods The techniques of immunochemistry and TRAP-ELISA were employed to detect the expression of hTERT and telomerase activity in different tumor cell lines.Results The positive rate of hTERT in HeLa cells(93.75%?3.10%)was significantly higher than that in U2OS cells(2.75%?0.96%),besides the other three cell lines showed an positive rates of hTERT in MCF-7(92.50%?2.65%),SMMC7721 (53.75%?2.22%)and PC-3m(23.50%?2.89%); Meanwhile,the telomerase activity of HeLa cells (94.58%?3.49%) was also much higher than that of U2OS(3.02%?0.43%),likewise the telomerase activities of the other three cell lines (MCF-7,SMMC7721,PC-3m) were 73.90%?4.50%,66.67%?3.35% and 50.62%?1.96%, respectively. Conclusion The expression of hTERT and telomerase activity show obvious differences among five tumor cell lines,suggesting that telomerase inhibitors cannot effect on all the tumor cell lines.
5.Isolation and purification of plasminogen activator of Yersinia pestis
Li-yuan, SHI ; Guo-lin, YU ; Li, BAI ; Guang-can, YANG ; Shan-shan, DONG
Chinese Journal of Endemiology 2009;28(4):365-368
Objective To find methods to isolate and purify plasminogen activator (Pla) from artificial culture of Yersinia pestis. Methods Ultrasonication and urea extracting combined by ammonium sulfate salting-out were tried to extract Pla. High performance liquid chromatography(HPLC) was used to purify Pla. The first step was ion exchange and the second was gel filtration, Preparative electrophoresis was used to purify Pla, too. The enzyme activity of the isolated or purificated Pla was detected. Results Both 50% - 60% saturated ammonium sulfate deposition of supernatant of plague bacilli ultrasonication and 0 - 10% saturated ammonium sulfate deposition of supernatant of plague bacilli powder soaked by urea had three bands(Mr about 31×103, 35×103 and 37×103) and lysis rings were 6.5 and 7.2 mm in diameter respectively when the enzyme activity was detected. Pla purified by HPLC was mainly composed of three bands(Mr about 31×103, 35×103 and 37×103), occupying more than 80% of total protein weight and lysis ring was 5.0 mm in diameter. Pla purified by preparative electrophoresis mainly consisted of three bands(Mr about 31×103, 35×103 and 37×103) with other proteins of low concentration nearby, no lysis ring was detected. Conclusions Pla is collected by the methods of ultrasonication and urea extracting. Priliminary purification of Pla can be achieved by HPLC and preparative electrophoresis.
6.PI_3 K/Akt signal effects Ephrin-A1 mediated malignancy of HCC cells
Gang CHEN ; Yi WANG ; Mengtao ZHOU ; Hongqi SHI ; Zhengping YU ; Yefan ZHU ; Fuxiang YU ; Yunfeng SHAN
Chinese Journal of General Surgery 2009;24(10):788-791
Objective To investigate the role of PI_3 K/Akt signal pathway in Ephrin-Al gene mediated invasion,metastasis of Huh-7 cells.Methods Western blot was used to test the protein expression of phosphatidylinositol 3-kinase(PI_3 K)and mitogen-activated protein kinase(MAPK)after Huh-7 cells were treated with Ephrin-A1/Fc fusion protein.According to the protein expression,LY294002 was used to block PI_3 K/Akt pathway specifically,then p-Akt protein expression,mobility and invasive ability of Huh-7 cells were examined.Results In Huh-7 cells actived by Ephrin-Al/Fc fusion protein,p-Akt expression was higher than that in control group(t=4.564,P<0.05),but there was no difference of p-p38MAPK expression between Ephrin-Al/Fc fusion protein group and IgG/Fc fusion protein group(P>0.05).PI_3 K/Akt pathway was specifically blocked by LY294002,the p-Akt protein expression decreased in Huh-7 cells,and the mobility and invasive ability mediated by Ephrin-Al in Huh-7 cells decreased(P<0.05).Conclusions PI_3 K/Akt pathway effects an important role in mobility and invasive ability of Huh-7 cells mediated by Ephrin-A1.
7.Application and value of fluorescence quantitative PCR in detecting Mycobacterium tuberculosis in paraffin embedded specimens.
Chun-ying LUO ; Jian-dong WANG ; Xuan WANG ; Heng-hui MA ; Shan-shan SHI ; Bo YU ; Xiao-jun ZHOU
Chinese Journal of Pathology 2012;41(8):562-563
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Female
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Fluorescence
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Follow-Up Studies
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Humans
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Male
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Middle Aged
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Mycobacterium tuberculosis
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isolation & purification
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Paraffin Embedding
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Polymerase Chain Reaction
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methods
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Retrospective Studies
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Tuberculosis, Pulmonary
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diagnosis
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microbiology
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Young Adult
8.The interactions between natural products and OATP1B1.
Mei-zhi SHI ; Yu LIU ; Jia-lin BIAN ; Meng JIN ; Chun-shan GUI
Acta Pharmaceutica Sinica 2015;50(7):848-853
Organic anion transporting polypeptide 1B1 (OATP1B1) is an important liver-specific uptake transporter, which mediates transport of numerous endogenous substances and drugs from blood into hepatocytes. To identify and investigate potential modulators of OATP1B1 from natural products, the effect of 21 frequently used natural compounds and extracts on OATP1B1-mediated fluorescein methotrexate transport was studied by using Chinese hamster ovary cells stably expressing OATP1B1 (CHO-OATP1B1) in 96-well plates. This method could be used for the screening of large compound libraries. Our studies showed that some flavonoids (e.g., quercetin, quercitrin, rutin, chrysanthemum flavonoids and mulberrin) and triterpenoids (e.g., glycyrrhetinic acid and glycyrrhizic acid) were inhibitors of OATP1B1 with IC50 values less than 16 µmol · L(-1). The IC50 value of glycyrrhetinic acid on OATP1B1 was comparable to its blood concentration in clinics, indicating an OATPlB1-mediated drug-drug interaction could occur. Structure-activity relationship analysis showed that flavonoids had much higher inhibitory activity than their glycosides. Furthermore, the type and length of saccharides had a significant effect on their activity. In addition, we used OATP1B1 substrates fluvastatin and rosuvastatin as probe drugs to investigate the substrate-dependent effect of several natural compounds on the function of OATP1B1 in vitro. Our results demonstrated that the effect of these natural products on the function of OATPlB1 was substrate-dependent. In summary, this study would be conducive to predicting and avoiding potential OATP1B1-mediated drug-drug and drug-food interactions and thus provide the experimental basis and guidance for rational drug use.
Animals
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Biological Products
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CHO Cells
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Cricetulus
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Drug Interactions
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Fatty Acids, Monounsaturated
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pharmacology
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Flavonoids
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pharmacology
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Indoles
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pharmacology
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Inhibitory Concentration 50
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Organic Anion Transporters
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genetics
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metabolism
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Rosuvastatin Calcium
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pharmacology
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Structure-Activity Relationship
9.Comparison and application of three visual rating scales for white matter lesions in Alzheimer's disease and mild cognitive impairment
Wanxin MA ; Shan SHI ; Jing LIAO ; Huali WANG ; Xin YU ; Huishu YUAN
Chinese Journal of Geriatrics 2013;32(10):1042-1046
Objective To compare the clinical value of three visual rating scales (VRS) for white matter lesions (WML) in Alzheimer's disease (AD) and mild cognitive impairment (MCI).Methods Totally 184 subjects (including 107 AD patients,47 MCI patients and 30 normal controls)were recruited.All subjects underwent comprehensive neuropsychological tests and were examined with a standard protocol of MR imaging.WML burden was rated with the Age-Related White Matter Changes (ARWMC) rating scale,Cholinergic Pathways Hyperintensities Scale (CHIPS) and Fazekas scale.Consistence of the three rating scales were analyzed,and detection results of WML in AD/ MCI/NC group with the three rating scales were compared.The relationship between WML and cognitive function in AD and MCI groups were explored.Results ARWMC rating scale,CHIPS and Fazekas scales were moderately to highly correlated with WML (r=0.61-0.78,all P<0.01).ARWMC showed that WML were in the left and right frontal lobes in AD and MCI groups and in the right frontal lobe and left temporal lobe in NC group,which had significant differences (F=4.20,4.69,4.69,3.36,respectively,all P<0.05).WML in the left temporal lobe had a significant difference between MCI group and NC group (F=3.36,P<0.05).CHIPS showed that WML in the front left side of centrum semiovale in AD and MCI groups had significant difference as compared with that in NC group (F=4.88,P<0.05),and WML in the rear right side of low external capsule had a significant difference between AD and NC groups (F=3.04,P<0.05).Fazekas scale showed that there was no difference in WML among AD,MCI and NC groups.ARWMC showed that scores of WML in right temporal lobe were positively correlated with concentration and visual construction,and bilateral basal ganglia were negatively correlated with the concentration.Fazekas scale showed that scores of WML in periventricular were negatively correlated with concentration,MMSE and orientation,scores of WML in deep white matter were negatively correlated with concentration,abstraction and judgment,and the whole scores were negatively correlated with concentration.CHIPS showed that the scores of WML in the rear left side of low external capsule were negatively correlated with short-term memory in MCI group.Conclusions The three rating scales well consistent with each other.ARWMC rating scale and CHIPS can detect the differences in WML between AD,MCI and NC groups.The scores of CHIPS in WML are related with cognition in MCI group,while scores of ARWMC and Fazekas scale in WML are correlated with many aspects in cognitive function in AD group.In general speaking,CHIPS in MCI patients and ARWMC rating scale in AD patients show more advantages in the detection of WML and have better correlation with cognitive function.
10.Study on induction of immunotolerance against Graves' disease in neonatal mice
Liping WU ; Liru XUN ; Bingyin SHI ; Jing YANG ; Li XU ; Zhufang TIAN ; Shan GAO ; Yu ZHANG
Chinese Journal of Endocrinology and Metabolism 2012;28(9):744-749
Objective To investigate the feasibility of inducing neonatal immunotolerance against Graves'disease by gene TSH receptor (TSHR) 289 and its possible mechanism.Methods Neonatal (0-24 h) female BALB/c mice were divided into intraperitoneal injection group,intramuscular injection group,model group,and normal control group.The intraperitoneal group and the intramuscular group were further divided into low-dosage,middle-dosage,high-dosage tolerance groups,and the coresponding control groups.The tolerance groups and the controls were intraperitoneally or intramuscularly pretreated with low-dosage( 1×106 particles),middle-dosage( 1 × 108particles),high-dosage( 1 × 1010 particles)of Ad-TSHR 289 or Ad-lacz respectively.6 to 7 weeks later,the normal control group received intramuscular injection with Ad-lacz; the other groups were immunized with Ad-TSHR289,three times at 3 weeks interval.10 days after the first immunization,serum TRAb was detected.4 weeks after the last immunization,serum TRAb,TT4,splenic CD4 + CD25 + Foxp3/CD4 + were tested,and the thyroid tissues were examinated histologically.Results Ten days after the first immunization,no antibody response against TSHR was detected in the two high-dose tolerance groups,but the TRAb titer in respective controls was significantly higher( P<0.05 ).4 weeks after the last injection,in high-dose tolerance groups,only 1/10 of mice immunized by intraperitoneal or intramuscular injection elicited anti-TSHR antibody,and no mice immunized intraperitoneally had elevated serum TT4.Two of ten mice challenged intramuscularly showed slightly increased TT4 levels,but the respective controls displayed a strong antibody response( P<0.01 ) and elevated TT4 level ( P<0.05 ).The similar percentages of high TT4 and thyroid hyperplasia were found in all groups.Additionally,the frequencies of CD4+CD25 +Foxp3/CD4+in two high-dose tolerance groups were significantly increased as compared to those in controls( P<0.05 ).The incidence of Graves' disease in the other groups by intraperitoneal or intranuscular injections was not statistically different from those in the corresponding control groups and the model group.Conclusions The immune tolerance against Graves'disease is induced in neonatal mice by either intraperitoneal or intramuscular pathway with specific antigen of TSHR 289,carried by adenovirus vector,and then inhibits Graves' disease in adults. Stimulation with the high-dosage antigen is liable to induce immune unresponsiveness.CD4 + CD25 + Foxp3 +T cells may play an important role in the induction and maintenance of tolerance.