2.Method for studying traditional Chinese compound prescription
Jianyang XU ; Liwu LIANG ; Dewen QIU ; Xiao HAN
Journal of Integrative Medicine 2005;3(5):347-50
Treatment with traditional Chinese medicinal composite is one of the most important characteristics of traditional Chinese medicine (TCM). Studying material base of TCM composite and its mechanism is a key to modernizing the industry of Chinese medicinal herbs. The research for TCM composite can be carried out from many different angles, including multiple components, multiple actions, multiple levels and multiple targets. Such a way to study TCM composite will be beneficial to improving the theory of TCM composite, guiding clinical administration and developing new products.
5.Behavior Characteristics in Children with Benign Epilepsy Combined with Centro-Temporal Spikes
yu-han, XIAO ; si-yuan, YANG ; hui-ci, LIANG
Journal of Applied Clinical Pediatrics 2004;0(12):-
Objective To explore the behavior characteristics in children with benign epilepsy combined with centro-temporal spikes(BECTS).Methods Eighty-two children with BECTS aged 2.5-3.0(2.65?2.31)years old,51 male,31 female,who were free of mental retardation assessed with Gesell developmental schedules,untreated with antiepileptic drugs,and were investigated 15 days after the latest seizure.Eighty-two healthy children with sex and age matched to the cases,53 male,29 female,aged 2.5-3.0(2.6?0.4)years old.The behavior characteristics of infants in BECTS group and control group were assessed with CBCL,including 6 behavior factors which were sleep problem,social flinches,depression,physical aspect,attacking,act of sabotage and the infants-middle school student social ability scale.Results The total scores of behavior characteristics and the scores of depression,sleep problem,attacking and act of sabotage in BECTS group were all higher than those in control group,the differences were statistically significant.However,scores of social flinches,and physical aspect in BECTS group had no significant differences compared with those of control group.There were no significant difference of social adaptive component between the BECTS and control group.Conclusions Children with BECTS have behavior disorders to some extent,but their social adaptive capacity are the same as normal children.
6.Cloning, prokaryotic expression, and functional identification of a sesquiterpene synthase gene (AsSS4) from Aquilaria sinensis.
Liang LIANG ; Qing-Mei GUO ; Zheng ZHANG ; Yan-Hong XU ; Xiao-Min HAN ; Juan LIU
Acta Pharmaceutica Sinica 2014;49(12):1724-1729
A sesquiterpene synthase (AsSS4) full-length open reading frame (ORF) cDNA was cloned from wounded stems of Aquilaria sinensis by RT-PCR method. The result showed that the ORF of AsSS4 was 1,698 bp encoding 565 amino acids. Prokaryotic expression vector pET28a-AsSS4 was constructed and transformed into E. coli BL21 (DE3) pLysS. Recombinant AsSS4 protein was obtained after induction by IPTG and SDS-PAGE analysis with a MW of 64 kD. Enzymatic reactions using farnesyl pyrophosphate showed that recombinant AsSS4 protein purified by Ni-agarose gel yielded five sesquiterpene compounds, cyclohexane, 1-ethenyl-1-methyl-2, 4-bis(1-methylethenyl)-, β-elemene, α-guaiene, α-caryophyllene and δ-guaiene. This paper reported the first cloning and functional characterization of AsSS4 gene from A. sinensis, which will establish a foundation for future studies on the molecular mechanisms of wound-induce agarwood formation in A. sinensis
Alkyl and Aryl Transferases
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biosynthesis
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genetics
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Azulenes
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Cloning, Molecular
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DNA, Complementary
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Escherichia coli
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Open Reading Frames
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Polyisoprenyl Phosphates
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Recombinant Proteins
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biosynthesis
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Sesquiterpenes
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metabolism
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Sesquiterpenes, Guaiane
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Thymelaeaceae
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enzymology
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genetics
7.Cloning and expression analysis of cinnamate 4-hydroxylase (C4H) reductase gene from Aquilaria sinensis.
Liang LIANG ; Xiao-Min HAN ; Zheng ZHANG ; Qing-Mei GUO ; Yan-Hong XU ; Juan LIU ; Yong-Cui LIAO
China Journal of Chinese Materia Medica 2014;39(10):1767-1771
The study aimed to clone the open reading frame of cinnamate 4-hydroxylase (C4H) from Aquilaria sinensis and analyze the bioinformatics and expression of the gene. One unique sequence containing C4H domain was discovered in our previous reported wound transcriptome dataset of A. sinensis. The open reading frame of C4H was cloned by RT-PCR strategy with the template of mixed RNA extracted from A. sinensis stem which treated by different wound time. The bioinformatic analysis of this gene and its corresponding protein was performed. C4H expression profiles in responds to MeJA (methyl jasmonate) application were analyzed by real-time PCR. The length of C4H open reading frame (ORF) was 1 515 bp, encoding 514 amino acids. The GenBank accession number is KF134783. Inducible-experiments showed that the genes were induced by mechanical wound as well as MeJA induction, and reached the highest expression level at 8 h and 20 h, respectively. The full-length cDNA of C4H and its expression patterns will provide a foundation for further research on its function in the molecular mechanisms of aromatic compounds and flavonoids biosynthesis.
Amino Acid Sequence
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Cloning, Molecular
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Models, Molecular
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Molecular Sequence Data
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Open Reading Frames
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Oxidoreductases
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chemistry
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genetics
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metabolism
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Phylogeny
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Plant Proteins
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chemistry
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genetics
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metabolism
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Thymelaeaceae
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chemistry
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enzymology
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genetics
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Trans-Cinnamate 4-Monooxygenase
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chemistry
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genetics
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metabolism
8.Study of production of sesquiterpenes of Aquilaria senensis stimulated by Lasiodiplodia theobromae.
Xiao-Min HAN ; Liang LIANG ; Zheng ZHANG ; Xiu-Jin LI ; Yun YANG ; Hui MENG ; Zhi-Hui GAO ; Yan-Hong XU
China Journal of Chinese Materia Medica 2014;39(2):192-196
To investigate the mechanism of agarwood formation in Aquilaria sinensis induced by Lasiodiplodia theobromae, the fermentation liquor of L. theobromae was analyzed qualitatively and quantitatively by gas chromatography-mass spectrometry (GC-MS). JAs were detected in the fermentation liquor. The effect of the fermentation liquor on the abundance of sesquiterpenes in the callus of A. sinensis was analyzed by solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). And the fermentation liquor stimulated alpha-guaiene, alpha-humulene and delta-guaiene biosynthesis in calli. It was inferred that L. theobromae produced JAs, which resulted in a significant increase of sesquiterpenes in A. sinensis.
Ascomycota
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physiology
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Fermentation
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Sesquiterpenes
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metabolism
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Thymelaeaceae
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metabolism
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microbiology
9.The relationship between CHADS2 score and prognosis in acute ischemic stroke patients with nonvalvular atrial fibrillation
Liangtong HUANG ; Zhao HAN ; Zusen YE ; Hongfei JING ; Zheng ZHANG ; Zhen WANG ; Liang FENG ; Meijuan XIAO
Chinese Journal of Neurology 2012;45(3):169-173
Objective To explore the relationship between the CHADS2 score and short-term prognosis in acute ischemic stroke patients with nonvalvular atrial fibrillation. Methods Consecutive ischemic stroke patients with nonvalvular atrial fibrillation who were hospitalized within 7 days after stroke were registered.Patients were divided into 3 groups on the basis of CHADS2 pre-stroke score (0 to 1,2,3to 6).And recovery was assessed by modified Rankin Scale (mRS) at 3 months follow-up (mRS ≤ 2reflected good prognosis,and mRS ≥ 5 implicated unfavorable outcome).After screening the risk factors affecting prognosis using univariate analysis,the relationship between CHADS2 score and prognosis was estimated using logistic regression model.Results Of 203 patients enrolled,the CHADS2 score of 0-1,2,3-6 were present in 72,53 and 78 respectively.Patients with the higher CHADS2 score had a higher initial NIHSS score (9.8,12.6,13.0,F =3.404,P =0.008 ),more likely to receive nosocomial pulmonary infection( 12.5%,34.0%,39.7%,x2 =14.643,P =0.001 ),negatively related to good prognosis (x2 =28.542,P =0.000 ) and positively related to unfavorable outcome ( x2 =23.575,P =0.000 ).Logistic regression model results suggested that CHADS2 score was an independent predictor of good prognosis and unfavorable outcome. Compared with CHADS2 score 3-6, CHADS2 score 0-1 was independently associated with good prognosis ( OR =5.018,95 % CI 2.055-12.560 ).And compared with CHADS2 score 0-1,CHADS2 score 3-6 was independently associated with unfavorable outcome ( OR =6.197,95% CI1.670-22.996 ).Conclusions CHADS2 pre-stroke score appears to be useful in prediction of stroke outcome for patients with nonvalvular atrial fibrillation.After acute ischemic stroke,the patients with the lower CHADS2 score (0-1) are more likely to achieve good prognosis,inversely,the patients with the higher CHADS2 score (3-6) are more likely to achieve unfavorable outcome.
10.Relationship between MTA1 expression and invasive and metastatic ability of cervical cancer cell
Xiaoyan HAN ; Haili QIAN ; Junjun YANG ; Xueyan ZHANG ; Ming FU ; Xiao LIANG ; Chen LIN ; Yang XIANG
Chinese Journal of Obstetrics and Gynecology 2011;46(9):678-683
ObjectiveTo investigate the relationship between metastasis-associated gene 1 ( MTA1 )expression and invasive and metastatic ability of cervical cancer cell. MethodsThree kinds of plasmids pcDNA3( control group), pcDNA3-MTA1 ( MTA1 group) and pSilencer3. 1-MTA1-siRNA ( MTA1-siRNAgroup) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability. ResultsCompared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P<0. 05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ±6, 169 ± 10 and 57 ±5,respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group(P < 0. 05 ). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69. 3 ± 3. 6) %, ( 80. 4 ± 5. 6 ) % and ( 39. 2 ± 7.4 ) % separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P <0. 05 ). In the migration assay, the number of cells migrated to the bottom side of the membrane in control,MTA1 and MTA1-siRNA group were 153 ± 17,247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19,354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA 1-siRNA group showed lower cell migration and invasion ability (P < 0. 05 ). In cell cycle experiment, no significant differences of cell proportions including G1, S and G2 stage were found among three groups (P > 0.05).In animal experiment, compared with control group,MTA1 group showed accelersted tumor formation and growth,whilethe MTA1-siRNA group showed the reverse effect ( P < 0. 05 ). ConclusionsMTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.