1.The Establishment of an Anti-Trypanosoma Drug Screening System with Leucyl-tRNA Synthetase as an Inhibition Target
Guang-Wei GAO ; Ying YAO ; Da-Zhong DING ; Long YE ; Hu-Chen ZHOU ; Da-Wei LI ;
China Biotechnology 2006;0(12):-
Trypanosoma is a human parasite severely affecting poor tropical areas.However,current frontline drugs for Trypanosoma treatment have severe side-effects with decreased effectiveness.Based on the fact that aminoacyl-tRNA synthetase is a bonafide drug target for several microorganisms,including bacteria and fungi,it is plausible that it may also be effective target of Trypanosoma.The Trypanosoma brucei leucyl-tRNA synthetase(tbLeuRS)was cloned,expressed and purified to develop an in vitro enzymatic assay system.The assay conditions were further optimized for the effective screening of tbLeuRS inhibitors thus establishing an anti-Trypanosoma drug screening system targeting tbLeuRS.The results indicated that this system can be employed for the effective screening of anti-Trypanosoma drugs with satisfactory specificity.In addition,this system can also be used for compound optimization,as well as IC50 testing.Using this system a series of compounds are identified that are effective Trypanosoma inhibitors without toxicity to human cells.Therefore,targeting tbLeuRS may represent a new venue for the development of anti-Trypanosoma drugs.
4.Changes of Atrial Natriuretic Peptide and Parathyroid Hormone in Umbilical Cord Blood of Newborn Infants with Intrauterine Growth Retardation and Their Relationships with Electrolytic
bing, HU ; wei, ZHOU ; hua-zhu, HU ; jin-ping, HU ; hei-da, LI
Journal of Applied Clinical Pediatrics 1986;0(02):-
Objective To explore the changes of atrial natriuretic peptide(ANP)and parathyroid hormone(PTH)in umbilical cord blood of newborn infants with intrauterine growth retardation(IUGR)and their relationships with electrolytic.Methods A total of 71 IUGR infants borned between Jan.2006 and Aug.2007 were enrolled in this study.Another 40 normal appropriate for gestational age neonates were selected as control group.The study group were divided into 2 groups:mature IUGR group(n=29)and premature IUGR group(n=42).The samples of umbilical cord blood of every group were collected at the time of delivery,and ANP,PTH levels in umbilical cord blood were mea-sured by radioimmunoassay.The sodium,calcium levels in their peripheral vein were measured simultaneously.Results 1.Compared with control group[(0.78?0.42)?g/L],the ANP levels of premature IUGR group[(1.26?0.47)?g/L] and the mature IUGR group[(1.09?0.51)?g/L] were significantly increased(t=5.98,2.76 Pa0.05).The calcium levels of the premature IUGR group[(1.85?0.37)mmol/L]significantly decreased(t=1.93 P0.05)compared with control group [(2.02?0.44)mmol/L].3.The serum sodium level was negatively correlated with the umbilical ANP level(r=-0.93 P
5.Studies on chemical constituents in fresh fleshy scaleleaf of Lilium lancifolium.
Wen-yan HU ; Jin-ao DUAN ; Da-wei QIAN ; Da-wei WANG
China Journal of Chinese Materia Medica 2007;32(16):1656-1659
OBJECTIVETo study the chemical constituents in fresh fleshyscaleaf of Lilium lancifolium.
METHODThe constituents were separated. by various kinds of chromatography and their structures were identified on the basis of spectral analysis.
RESULTTen compounds were identified regaloside A (1), regaloside C (2), methyl-a-D-mannopyranosid (3), methyl-ca-D-glucopyranoside (4), (25R, 26R) -26-methoxyspirost-5-ene-3p-yl-O-ca-L-rhamnopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (5), (25R)-spirost-5-ene-3beta-yl-O-alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (6), (25R, 26R)-17alpha-hydroxy-26-methoxyspirost-5-ene-3beta-yl-O-alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-glucopyra nosyl-(1-->6)]-beta-D-glucopyranoside (7), daucosterol (8), adenoside (9), berberine (10).
CONCLUSIONAll compounds except 1 and 3 were isolated from this species for the first time, and berberine was first reported in genus Lilium.
Berberine ; chemistry ; isolation & purification ; Lilium ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification
6.Isolation and differential expression of a novel MAP kinase gene DoMPK4 in Dendrobium officinale.
Gang ZHANG ; Yi-Min LI ; Ben-Xiang HU ; Da-Wei ZHANG ; Shun-Xing GUO
Acta Pharmaceutica Sinica 2014;49(7):1076-1083
Mitogen-activated protein kinases (MAPKs) are important signaling transduction components well conserved in eukaryotes and play essential roles in various physiological, developmental and hormonal responses in plant. In the present study, a MAPK gene, designated as DoMPK4 (GenBank accession No. JX297597), is identified from a rare endangered medicinal orchid species D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. The full length cDNA of DoMPK4 is 1 518 bp in length and encoded a 369 aa protein with a molecular weight of 42.42 kD and an isoelectric point of 5.55. DoMPK4 protein contained a serine/threonine protein kinase active site (158-170), a MAP kinase site (71-174), and eight conserved motifs. DoMPK4 had a transmembrane (214-232) but no signal peptide. Multiple sequence alignment showed that DoMPK4 shared high identities (74.9%-80.6%) with MAPK proteins from various plants. Phylogenetic analysis demonstrated that DoMPK4 belonged to group A of the MAPK evolutionary tree, and is closely related to monocots. Real time quantitative PCR (qPCR) analysis revealed that DoMPK4 is differentially expressed among the five organs including leaf, stem, root, seed, and protocorm-like body (PLB). The transcription level of DoMPK4 is the highest in the PLBs with 17.65 fold, followed by seeds, roots, and stems with 5.84, 2.28, and 1.64 fold, respectively. The progressive enhancement of DoMPK4 transcripts in the developing PLBs compared to that in the germinating seeds, suggests a role of DoMPK4 during the development of embryogenic PLBs formation in D. officinale.
Amino Acid Sequence
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DNA, Complementary
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genetics
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DNA, Plant
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genetics
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Dendrobium
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enzymology
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genetics
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Gene Expression Regulation, Plant
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Mitogen-Activated Protein Kinases
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genetics
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metabolism
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Phylogeny
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Plant Leaves
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metabolism
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Plant Proteins
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genetics
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metabolism
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Plant Roots
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metabolism
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Plant Stems
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metabolism
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Seeds
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metabolism
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Sequence Alignment
7.Usefulness of Ankle Brachial Index to Predict the Extent of Coronary Stenosis
Shi-wei YANG ; Da-yi HU ; Xian WANG ; Shouyan ZHANG ; Jian ZHANG ; Shi GUO
Chinese Journal of Rehabilitation Theory and Practice 2006;12(6):461-463
ObjectiveTo examine the relationship between ankle brachial index (ABI) and the extent of coronary stenosis and evaluate the usefulness of ABI to predict the extent of coronary stenosis in old patients.Methods118 patients with coronary angiography were examined by ABI and hemostatic factors evaluation in addition to history collection.ResultsABI was inversely and significantly associated with Gensini score. ABI reduced significantly (P<0.001) in the patients with 3-vessel or left main coronary artery disease (CAD). But there were no significant differences in ABI among the patients with no CAD, 1-vessel or 2-vessel CAD. The corresponding area under the ROC curve was 0.75±0.045, with 95% CI=0.67~0.84 (P<0.001) in ABI in 3-vessel or left main CAD. When ABI≤0.9, it had a relatively high specificity (89.1%) and sensitivity (55.6%) for predicting the presence of 3-vessel disease or left main CAD.ConclusionIn the old patients, ABI is inversely and significantly associated with the extent of coronary stenosis, and ABI≤0.9 has a relatively high specificity and sensitivity for predicting the presence of 3-vessel or left main CAD.
8.Differential expression profile of microRNAs between human hepatocellular carcinoma cell line and human normal hepatocyte cell line
Jiaxiang WANG ; Yuan WEI ; Qian HU ; Qiuliang LIU ; Yingzhong FAN ; Heying YANG ; Suning LI ; Da ZHANG ; Qian ZHANG
Chinese Journal of Hepatobiliary Surgery 2011;17(6):492-496
Objective To establish the expression profiles of microRNA (miRNA) in SMMC-7721 and CCC-HEL-1 cell lines in order to provide new clues to study the mechanism of miRNA function in hepatocellular carcinoma (HCC) and its treatment. Methods SMMC-7721 and CCC-HEL-1 cell lines were cultured in vitro. Total RNAs were extracted by TRIzol, followed by RNA quantification and quality control. The RNAs were used to detect the expression of miRNA in these two cell lines by miRNA array. The miRNA of interest was then verified by real-time PCR. Results In total,238 differentially expressed miRNAs were found, including 154 overexpressed and 84 underexpressed miRNAs. 64 miRNAs were upregulated more than 4 times and26 miRNAs were upregulated more than 10 times. 22 miRNAs were downregulated more than 4 times 10 miRNAs weredownregulated more than 5times, and 1 miRNA was downregulated more than 10 times. Real-time PCR validation suggested that has-miR-205 and has-let-7f in liver cancer increased by 2. 7 and 2. 3 times, respectively.Conclusion There are differences in the expression of miRNA between the SMMC-7721 and CCC-HEL-1 cell lines and the profiles of miRNA expression were established for both cell lines. It was found that has-miR-205 and has-let-7f were upregulated in liver cancer.
9.The expression of interferon-?in the peripheral blood and correlation with disease activity in systemic lupus erythematosus patients
Shan DENG ; Da-Wei HU ; Yuan WANG ; Nan SHEN ; Yue-Ying GU ; Chun-De BAO ; Shun-Le CHEN ;
Chinese Journal of Rheumatology 2003;0(11):-
Objective To investigate the mRNA and protein expression levels of interferon(IFN)-?in the peripheral blood of patients with systemic lupus erythematosus(SLE),to analyze the relationship between IFN-?and disease activity,and to evaluate the role of IFN-?in the pathogenesis of lupus.Methods SYBR green dyeⅠbased real-time quantatives PCR method was used to compare the mRNA expression levels of IFN-?in the peripheral blood leucocyte of SLE patients and healthy controls.Surum levels of IFN-?were measured with ELISA method.Results IFNA1 mRNA expression level in SLE patients(2.8?3.5)was signifi- cantly lower than that of normal controls(12.7?10.7,P=0.000).There was no significant difference between patients treated with glucocorticoid and those without in the expression level of IFNA1(P=0.549).Serum levels of IFN-?in SLE patients was significantly higher than that of normal controls(P=0.003).The SLEDAI score and anti-dsDNA antibody correlated positively,and complement components C3,C4 and leukocytes correlated negatively with serum concentration of IFN-?.IFN-?level correlated with the presence of fever and rash. Conclusion The close relationship between IFN-?serum level and disease activity in SLE patients suggests that IFN-?might be of importance in the disease process.
10.The expression and significance of glucose-6-phosphate isomerase mRNA in peripheral blood mononuclear cells of patients with rheumatoid arthritis
Guan-Fei ZHAO ; Yong-Zhe LI ; Yu-Guo SONG ; Chao-Jun HU ; Da-Wei TONG ; Shu-Lan ZHANG ;
Chinese Journal of Rheumatology 2001;0(04):-
Objective To investigate the mRNA level of glucose-6-phosphate isomerase(GPI)ex- pression in patients with rheumatoid arthritis(RA).Method Reverse transcription-polymerase chain reaction (RT-PCR)was applied to semiquantitatively analyze GPI mRNA expression in the peripheral blood mononu- clear cells(PBMCs)of 30 active RA patients,30 stable RA patients,30 patients with other rheumatic disease and 30 healthy subjects.Results There was statistically significant differences between patients with RA and other rheumatic diseases(P