3.Eexpression of Foxp3 and CD4~+CD_(25)~+ Regulation T Cells in Children with Acute Idiopathic Thrombocytopenic Purpura
xue-wen, LV ; fang, LIU ; chang-lin, WU
Journal of Applied Clinical Pediatrics 1993;0(03):-
Objective To investigate the amount of CD4+CD_(25)~+ regulation T cells(Tr cells) and the expression of Foxp3 in peripheral blood in children with acute idiopathic thrombocytopenic purpura(AITP),and analyze the relationship between CD4+CD_(25)~+ Tr cells and the immunopathogenesis of AITP.Methods CD4+CD_(25)~+ Tr cells were detected by flow cytometry in peripheral blood from AITP children and health children.Besides,related cytokine in peripheral blood were assigned by enzyme linked immunosorbent assay(ELISA).The expression of Foxp3 were detected by reverse transcriptase-polymerase chain reaction(RT-PCR).Results CD4+CD_(25)~+ Tr cells decreased obviously in peripheral blood in children compared with that of control group [(2.83?1.05)% vs (5.07?0.59)%,P
4.Expression of glutathione S-transferase mRNA in peripheral blood of the population in coal-burning fluorosis area
Chang-xue, WU ; Ke-ren, SHAN ; Zhi-zhong, GUAN
Chinese Journal of Endemiology 2010;29(2):121-123
Objective To investigate expression of glutathione S-transferase(GST) mRNA in peripheral blood of the population in coal-burning fluorosis area and to evaluate the effect of comprehensive control intervention. Methods Fifty samples of peripheral blood from patients in the coal-buring fluorosis area in Bijie county of Guizhou province were selected as fluorasis group and 50 samples of peripheral blood from patients in area with comprehensive management were selected as intervention group, respectively. Fifty samples from non-endemic fluorosis area were selected as the control group. Total RNA from blood was extracted and purified by the Trizol- Phenol-Chloroform one-step method. Expression of GST mRNA was detected by using SYBR Green I real-time fluorescence quantitative PCR. Results The data of GST mRNA in fluorosis group, intervention group and control group was 38.28±27.22,70.56±37.23 and 103.46 ± 46.62, respectively. There was a significant difference between the groups(F = 3.75, P < 0.05). Decreased expression of GST mRNA in fluorosis group and intervention group as compare to control was detected(all P < 0.05), and the expression of GST mRNA in intervention group was higher than that in fluorosis group(P < 0.05). Conclusion Coal-burning fluorosis possibly led to the decreased expression of GST mRNA in peripheral blood, and comprehensive control maybe prevent the decreased expression of GST in mRNA level.
5.Significance of Protein Kinase C Activity Detection in Children with Chronic Idiopathic Thrombocytopenic Purpura
chang-lin, WU ; jian-cheng, XUE ; xue-min, ZHOU ; fang, LIU
Journal of Applied Clinical Pediatrics 1993;0(03):-
Objective To investigate the changes of protein kinase C(PKC)activity in peripheral blood T lymphocytes in children with chronic idiopathic thrombocytopenic purpura(CITP)and the relationship between PKC activity,T lymphocytes activation and thrombocytes decrease.Methods Collecting sterile peripheral blood from CITP children(n=30)and healthy children(n=30),T lymphocytes were isolated and purified by the T cell segregation enrichment column,the PKC activity was detected by non-radioactive assay.Soluble interleukin-2 receptor(sIL-2R),which was T cell activated marker,was determined by enzyme-linked immunoabsorbent assay(ELISA),platelet was counted by cell counting meter.Results Compared with healthy children,PKC activity was significantly enhanced in CITP children[(0.94?0.23)mmol/(min?L)vs(0.50?0.17)mmol/(min?L),t= 8.42 P
6.Effects of α3 neuronal nicotinic acetylcholine receptor on cell apoptosis and p38 MAPK signal transduction pathway in SH-SY5Y cells.
Xue-ling ZHANG ; Xiao-lan QI ; Jia-mou REN ; Chang-xue WU ; Zhi-zhong GUAN
Chinese Journal of Pathology 2013;42(2):116-120
OBJECTIVETo investigate the effects of α3 neuronal nicotinic acetylcholine receptor (nAChR) on apoptosis and p38 signal transduction pathway in SH-SY5Y cells and to assess the roles of α3 nAChR in the pathogenesis of Alzheimer's disease (AD).
METHODSThe levels of α3 nAChR mRNA and protein were measured by real-time PCR and Western blot, respectively, in SH-SY5Y cells transfected with α3 nAChR siRNA. The mRNA level of bcl-2 and bax was measured by the real-time PCR. The siRNA transfected SH-SY5Y cells and control were then treated with 10 µmol/L Aβ25-35 for another 48 h, and the change in apoptotic rate and the levels of p-p38 and p38 were measured by flow cytometry and Western blot. Subsequently these SH-SY5Y cells were exposed to a blocker of p38 protein, and the apoptotic rate was measured again.
RESULTSCompared to the controls, the expression of α3 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with α3 nAChR siRNA decreased by 95% and 86%, respectively; the mRNA levels of bax increased 2.11 times and that for bcl-2 decreased 0.53 times. The apoptotic rate was unaffected (3.40% ± 0.20%); but it increased after Aβ25-35 treatment (24.52% ± 1.59%); the level of p-p38 protein also increased by 178% in the α3 nAChR inhibited cells treated with Aβ25-35. Compared to controls, the Aβ25-35-treated SH-SY5Y cells and the Aβ25-35-treated and siRNA-transfected cells both showed a reduction in apoptosis after treatment with p38 blocker, especially in the former.
CONCLUSIONThe siRNA silencing of α3 nAChR mRNA may enhance the effect of Aβ25-35 on the cell apoptosis by increasing the levels of p38 protein and bax mRNA and decreasing the level of bcl-2 mRNA, which may play a role in the pathogenesis of AD.
Alzheimer Disease ; etiology ; Amyloid beta-Peptides ; metabolism ; Apoptosis ; Cell Line, Tumor ; Gene Silencing ; Humans ; Neuroblastoma ; metabolism ; pathology ; Peptide Fragments ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Nicotinic ; genetics ; metabolism ; Signal Transduction ; Transfection ; bcl-2-Associated X Protein ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism
7.Chemical constituents of Swertia patens.
Kang HE ; Tuan-wu CAO ; Hong-ling WANG ; Chang-an GENG ; Xue-mei ZHANG ; Ji-jun CHEN
China Journal of Chinese Materia Medica 2015;40(20):4012-4017
Chemical constituents of Swertia patens. The whole plant of air-dried Swertia patens was extracted with 90% EtOH. The water extract was suspended in H₂O and extracted with petroleum ether, EtOAc and n-BuOH, successively. The compounds were isola- ted and purified by column chromatography from the EtOAc fraction, and identified based on spectral analyses (MS, ¹H-NMR, ¹³C- NMR). Eighteen compounds were isolated and elucidated as 3, 4-dihydro-1H,6H,8H-naptho [1,2-c:4,5-c', d'dipyrano-1, 8-dione (1), angelone (2), gentiogenal (3), erythricin (4), erythrocentaurin (5), gentianine (6), swertiakoside B (7), swertiamarin (8), 2'-O-actylswertiamarin (9), amarogentin (10), 1, 3, 5-trihydroxyxanthone (11), 1, 3-dihydroxy-5-methoxyxanthone (12), 1-hydroxy- 2, 3, 5-trimethoxyxanthone (13), gentiocrucine (14), 3-hydroxyphenylketone (15), n-hexacosyl ester 4-hydroxy-trans-cinnamate (16), n-hexacosyl ester 4-hydroxy-cis-cinnamate (17), and cholest-4-en-3-one (18). Compounds 1-7, 9-18 were obtained from S. patens for the first time.
Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Molecular Structure
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Spectrometry, Mass, Electrospray Ionization
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Swertia
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chemistry
9.Chemical constituents of Swertia angustifolia.
Kang HE ; Tuan-wu CAO ; Hong-ling WANG ; Chang-an GENG ; Xue-mei ZHANG ; Ji-jun CHEN
China Journal of Chinese Materia Medica 2015;40(18):3603-3607
This present work is to study the chemical constituents of Swertia angustifolia. The whole plants of air-dried Swertia angustifolia was extracted with 90% EtOH. The water extract was suspended in H2O and extracted with petroleum ether, EtOAc and nBuOH, successively. The compounds were isolated and purified by column chromatography from the EtOAc fraction, and identified based on spectral analyses (MS, 1H-NMR, 13C-NMR). Fourteen compounds were isolated and characterized as 1, 8-dihydroxy-3, 7-dimethoxyxanthone (1), 1, 8-dihydroxy-3, 5, 7-trimethoxyxanthone (2), 7-hydroxy-3, 8-dimethoxyxanthone-1-O-β-D-glucopyranoside (3), 8-0-[β-D-xylopyranosyl-(1-6) -β-D-glucopyranosyl] -1, 7-dihydroxy-3-methoxyxanthone (4), (+) -syringaresinol (5), ferulic acid (6), trans-coniferyl aldehyde (7), sinapaldehyde (8), trans-coniferyl alcohol (9), 3, 4-dihydroxybenzoic acid (10), 2-hydroxybenzoic acid (11), isophthalic acid (12), 2-furoic acid (13), and 2-methyl-4(3H)-quinazolinone(14). Compounds 2-14 were obtained from this plant for the first time.
Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Mass Spectrometry
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Molecular Structure
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Swertia
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chemistry
10.Chemical constituents of Swertia kouitchensis Franch.
Kang HE ; Tuan-wu CAO ; Hong-ling WANG ; Chang-an GENG ; Xue-mei ZHANG ; Ji-jun CHEN
China Journal of Chinese Materia Medica 2015;40(19):3811-3817
This study is to investigate the chemical constituents of Swertia kouitchensis. The whole plants of air-dried Swertia kouitchensis was extracted with 90% EtOH. The water extract was suspended in H2O and extracted with petroleum ether, EtOAc and n-BuOH, successively. The compounds were isolated and purified by column chromatography from the EtOAc fraction, and their structures were identified based on spectral analyses (MS, 1H-NMR, 13C-NMR). Twenty-eight compounds were obtained, and characterized as erythrocentaurin (1), erythrocentaurin dimethylacetal (2), swertiamarin (3), vogeloside (4), 2'-O- actylswertiamarin (5), swertianoside D (6), gentiocrucines A-B (7-8), gentiocrucine (9), 1-hydroxy-3, 7, 8-trimethoxyxanthone (10), 1-hydroxy-3, 5, 6-trimethoxyxanthone (11), 3-epitaraxerol (12), erythrodiol 3-O-palmitate (13), (+) -syringaresinol (14), caffeic acid (15), trans-coniferyl aldehyde (16), trans-coniferyl alcohol (17), 3, 4-dihydroxybenzoic acid (18), 4-hydroxy-3-methoxybenzoic acid (19), 3, 4-dihydroxybenzoic aldehyde (20), 2, 3-dihydroxybenzoic acid (21), 4-hydroxybenzoic acid (22), 3-acetoxybenzoic acid (23), 3-hydroxybenzoic acid (24), 3-hydroxybenzoic alcohol (25), nicotinic acid (26), 2-furoic acid (27), and uracil (28). Compounds 1-4, 6-28 were obtained from S. kouitchensis for the first time.
Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Magnetic Resonance Spectroscopy
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Mass Spectrometry
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Molecular Structure
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Swertia
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chemistry