1.Molecular characterization of a HMG-CoA reductase gene from a rare and endangered medicinal plant, Dendrobium officinale.
Lin ZHANG ; Ji-Tao WANG ; Da-Wei ZHANG ; Gang ZHANG ; Shun-Xing GUO
Acta Pharmaceutica Sinica 2014;49(3):411-418
The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes the conversion of HMG-CoA to mevalonate in mavalonic acid pathway, which is the first committed step for isoprenoid biosynthesis in plants. However, it still remains unclear whether HGMR gene plays a role in the isoprenoid biosynthesis in Dendrobium officinale, an endangered epiphytic orchid species. In the present study, a HMGR encoding gene, designed as DoHMGR1 (GenBank accession JX272632), was identified from D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods, for the first time. The full length cDNA of DoHMGR1 was 2 071 bp in length and encoded a 562-aa protein with a molecular weight of 59.73 kD and an isoelectric point (pI) of 6.18. The deduced DoHMGR1 protein, like other HMGR proteins, constituted four conserved domains (63-561, 147-551, 268-383 and 124-541) and two transmembrane motifs (42-64 and 85-107). Multiple sequence alignment and phylogenetic analyses demonstrated that DoHMGR1 had high identity (67%-89%) to a number of HMGR genes from various plants and was closely related to Vanda hybrid cultivar, rice and maize monocots. Real time quantitative PCR (qPCR) analysis revealed that DoHMGR1 was expressed in the three included organs. The transcripts were the most abundant in the roots with 2.13 fold over that in the leaves, followed by that in the stems with 1.98 fold. Molecular characterization of DoHMGR1 will be useful for further functional elucidation of the gene involving in isoprenoid biosynthesis pathway in D. officinale.
Base Sequence
;
Cloning, Molecular
;
DNA, Complementary
;
genetics
;
Dendrobium
;
enzymology
;
genetics
;
Gene Expression Regulation, Plant
;
Hydroxymethylglutaryl CoA Reductases
;
genetics
;
metabolism
;
Molecular Weight
;
Phylogeny
;
Plant Leaves
;
enzymology
;
genetics
;
Plant Roots
;
enzymology
;
genetics
;
Plant Stems
;
enzymology
;
genetics
;
Plants, Medicinal
;
enzymology
;
genetics
;
Sequence Alignment
;
Sequence Homology, Amino Acid
2.Primary liver paraganglioma: a case report.
Ping CHEN ; Ya-nan ZHAI ; Hong LIU ; Hao WANG ; Shun-lin GUO
Chinese Journal of Hepatology 2013;21(10):786-787
3.Study on chemical components distribution in Paeoniae Radix Alba and its processing methods.
Lin JIN ; Wan-shun ZHAO ; Qiao-sheng GUO ; Wen-sheng ZHANG ; Zheng-liang YE
China Journal of Chinese Materia Medica 2015;40(10):1953-1959
After studying the discipline of chemical components distribution in peony root, it was found that the content distribution of albiflorin, paeoniflorin, benzoylpaeoniflorin and paeonol was inhomogeneous in different parts of peony root, as same as in different roots belonging to a same plant. Therefore, in order to minimize the effects of different qualities of materials on experimental result, the content changing percentages of four chemical components was chosen as indicators to study processing method of Paeoniae Radix Alba. The processing technique of Paeoniae Radix Alba was put peony roots in the boiled water for at most ten minutes, and cut them into 2-3 mm slices after peeling the root bark, finally dry them in a oven under 55 degrees C, which was similar to the traditional processing method used in Bozhou. And the peeling, boiling and drying methods would affect the contents of albiflorin, paeoniflorin, benzoylpaeoniflorin and paeonol on several levels.
Chemistry, Pharmaceutical
;
methods
;
Chromatography, High Pressure Liquid
;
Drugs, Chinese Herbal
;
chemistry
;
isolation & purification
;
Paeonia
;
chemistry
;
Plant Roots
;
chemistry
4.Optimization of extraction technology from Paeoniae Radix Alba using response surface methodology.
Lin JIN ; Wan-shun ZHAO ; Qiao-sheng GUO ; Wen-sheng ZHANG ; Zheng-liang YE
China Journal of Chinese Materia Medica 2015;40(15):2988-2993
To ensure the stability of chemistry components and the convenience of operation, ultrasound method was chosen to study in this investigation. As the total common peaks area in chromatograms was set to be evaluation index, the influence on the technology caused by extraction time, ethanol concentration and liquid-to-solid ratio was studied by using single factor methodology, and the extraction technology of Paeoniae Radix Alba was optimized by using response surface methodology. The results showed that the extracting results were most affected by ethanol concentration; liquid-to-solid ratio came the second and extraction time thirdly. The optimum ultrasonic-assisted extraction conditions were as follow: the ultrasonic extraction time was 20.06 min, the ethanol concentration in solvent was 72.04%, and the liquid-to-solid ratio was 53.38 mL · g(-1), the predicted value of total common peaks area was 2.1608 x 10(8). Under the extraction conditions after optimization, the total common peaks area was 2.1422 x 10(8), and the relative deviation between the measured and predicted value was 0.86%, so the optimized extraction technology for Paeoniae Radix Alba is suitable and feasible. Besides, for the purpose of extracting more sufficiently and completely, the optimized extraction technology had more advantages than the extraction method recorded in the monogragh of Paeoniae Radix Alba in Chinese Pharmacopoeia, which will come true the assessment and utilization comprehensively.
Paeonia
;
chemistry
;
Technology, Pharmaceutical
;
methods
5.Determination of chemical components of Paeoniae Radix Alba decoction pieces and its quality evaluation.
Lin JIN ; Wan-shun ZHAO ; Qiao-sheng GUO ; Zheng-liang YE ; Wen-sheng ZHANG
China Journal of Chinese Materia Medica 2015;40(3):484-489
OBJECTIVEAn UPLC method was developed to evaluate the quality of Paeoniae Radix by simultaneously determining four components, paeoniflorin, albiflorin, benzoylpaeoniflorin and paeonol in Paeoniae Radix Alba decoction pieces.
METHODThe UPLC chromatographic column was ACQUITY UPLC® HSS T3. The mobile phase was acetonitrile-0.05% phosphoric acid water with gradient elution. The column temperature was 30 °C and detection wavelength was 230 nm with a flow rate of 0.4 mL · min(-1). A linear model was obtained through principal component analysis (PCA), and PCA scores were used to evaluate the quality of Radix Paeoniae Alba decoction pieces comprehensively.
RESULTPaeoniflorin, albiflorin, benzoylpaeoniflorin and paeonol could be well separated from other components, and the results of specificity, precision, repeatability, linearity, recovery rate and stability reached the standards, respectively. The content of paeoniflorin in 9 batches of Paeoniae Radix Alba decoction pieces was below the standard given by Chinese Pharmacopoeia (2010 edition). Using the comprehensive scoring method with principal component analysis, the results showed that the samples from Zhejiang province have better quality than those from Anhui and Shandong provinces.
CONCLUSIONThe method established in this study can effectively determine the content of paeoniflorin, albiflorin, benzoylpaeoniflorin and paeonol, which could be used for quality control of Paeoniae Radix Alba.
Drugs, Chinese Herbal ; analysis ; Paeonia ; chemistry ; Principal Component Analysis
6.Evaluation of the association between left coronary artery stenosis degree and myocardial perfusion by 64 multi-slice.
Chinese Journal of Cardiology 2012;40(8):672-675
OBJECTIVETo evaluate the association between left coronary artery stenosis degree and myocardial perfusion by 64 multi-slice CT.
METHODSA total of 223 patients underwent 64 multi-slice CT coronary artery images (CTA) were included and divided into normal group (91 cases), mild stenosis group (72 cases), moderate stenosis group (36 cases) and severe stenosis group (24 cases). Myocardial density was measured at apical, septal and lateral segments. Myocardial density in infarcted segments was compared to non-infarct segments in 11 patients with old myocardial infarction (all from severe stenosis group).
RESULTSMyocardial density was significantly lower at apical segments [(55.8 ± 21.4) HU vs. (75.3 ± 7.5) HU], at septal segment [(87.8 ± 3.3) HU vs. (98.2 ± 5.2) HU] and at lateral segment [(86.8 ± 7.9) HU vs. (95.6 ± 11.6) HU] in severe stenosis group than in normal group (all P < 0.05). Myocardial density of patients with old myocardial infarction was significantly reduced in non-infarct segment [(70.9 ± 8.3) HU vs.(98.7 ± 7.3) HU, P < 0.01] and increased in infarct segment [(42.5 ± 15.7) HU vs. (17.8 ± 4.1) HU, P < 0.01] post contrast enhancement.
CONCLUSIONCTA could be used to evaluate the severity of the left coronary artery stenosis based on myocardial density measurement. Myocardial delayed enhancement derived from CTA could be used to identify infarct segments.
Adult ; Aged ; Aged, 80 and over ; Coronary Angiography ; Coronary Stenosis ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnostic imaging ; Myocardial Reperfusion ; Tomography, Spiral Computed
7.Vascular effect of hydroxyl-safflor yellow A and underlying mechanism
lin, ZHANG ; guo-shun, SHEN ; jing-nian, ZHANG ; pie-he, NIE ; jian-ming, ZHI
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(04):-
Objective To investigate the vascular effect of hydroxyl-safflor yellow A(HSYA) on rat thoracic aorta and its underlying mechanism. Methods The tension of isolated thoracic aorta rings of rats perfused with different concentrations of HSYA(1?10-6-1?10-4 mol/L) was measured using organ bath technique.The effects of HSYA on the vasocontraction induced by cumulative phenylephrine(PE)(1?10-6-1?10-4 mol/L),KCl(6?10-2 mol/L) and CaCl2(1?10-5-3?10-3 mol/L) were recorded respectively. Results HSYAcaused a concentration-dependent anti-contraction effects by KCl or PE in endothelium-intact and endothelium-denuded aortic rings.HSYA inhibited the CaCl2-induced contraction and downward shifted concentration-response curve of aortic rings.HSYA+HP resulted in more significant anti-contraction effect than single use of HSYA(P0.05).There were significant differences in anti-contraction effect between HSYA+RR and RR or HSYA(P
8.The expression of interferon-regulatory factor genes in patients with systemic lupus erythematosus
Jie QIAN ; Nan SHEN ; Gui-Mei GUO ; Nian-Hong WAN ; Yan LIN ; Xin-Fang HUANG ; Hui WU ; Shun-Le CHEN ;
Chinese Journal of Rheumatology 2003;0(09):-
Objective To observe whether the expression of interferon-regulatory factor genes are re- lated to systemic lupus erythematosus (SLE).Methods The clinical data of 45 SLE patients and 37 normal controls were collected.Total RNA of peripheral blood was extracted and transcripted into cDNA.Sybr green dye based real-time quantitative PCR method was used to compare the expression (indicated as-??Ct value) of IRFI,IRF4,IRF8 in patients with SLE and those in the controls.Results The levels of IRF1,IRF4 and IRF8 mRNA were-3.90?0.19,-8.04?0.25 and 3.60?0.15 respectively in normal controls.In SLE patients, IRF4 mRNA expression was -8.82?0.18,higher than that in normal (P=0.011).But IRF8 mRNA expression was 3.09?0.13,lower than that in normal (P=0.012).Conclusion Abnormal IRF mRNA expression is found in the peripheral blood of SLE patients.IRFs may play roles in the pathogenesis of SLE by affecting the differen- tiation of Th cells.
9.Diterpene constituents of Tripterygium willfordii (II).
Sui LIN ; Hui-Qing QUE ; Hua-Yi PENG ; Li-Ping QIAN ; Shun-Min GUO ; Yuan-Chao LI
Acta Pharmaceutica Sinica 2011;46(8):942-945
In order to study the constituents and pharmacology of Tripterygium plants (Tripterygium willfordii Hook.f), a variety of chromatography methods were used. Four compounds were isolated from Tripterygium plant and their structures were elucidated by UV, IR, MS, HR-MS, 1H NMR, 13C NMR and 2D-NMR techniques. The isolated compounds were named as triptonide (1), neo-triptetraolide (2), 2alpha-hydroxytriptonide (3), and 15-hydroxytriptonide (4), separately. Compounds 3, 4 belong to new diterpenoids, which can inhibit the growth of K562 cells (leukemia cells) and HL60 cells (acute myeloid leukemia cells).
Cell Proliferation
;
drug effects
;
Diterpenes
;
chemistry
;
isolation & purification
;
pharmacology
;
Drugs, Chinese Herbal
;
chemistry
;
isolation & purification
;
pharmacology
;
HL-60 Cells
;
Humans
;
K562 Cells
;
Molecular Structure
;
Plant Roots
;
chemistry
;
Plants, Medicinal
;
chemistry
;
Tripterygium
;
chemistry
;
Triterpenes
;
chemistry
;
isolation & purification
10.A study of chronic hepatitis B infection superinfected with hepatitis E infection.
Guo-Shun ZHANG ; Fu-Min FENG ; Yu-Lin LI ; Ju-Xiang YUAN ; Hua SHANG
Chinese Journal of Hepatology 2006;14(12):906-908
OBJECTIVESTo compare the influence of hepatitis E virus (HEV) infection on the replication of hepatitis B virus (HBV) by observing clinical features, the relationship between the number of HBV DNA copies in serum, the degree of hepatic function impairments and prognosis of chronic hepatitis B patients with hepatitis E superinfection.
METHODSOne hundred and fifteen chronic hepatitis B (CHB) patients and 115 CHB patients with hepatitis E (HE) superinfection were studied and compared. 74 liver tissue biopsy samples of the CHB and 51 of the CHB-HE sufferers were obtained. HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc, anti-HBc-IgG, anti-HBc-IgM, HBV DNA and anti-HEVIgM were detected respectively by enzyme linked immunosorbent assay and polymerase chain reaction.
RESULTSThe number of HBV DNA copies in sera in the CHB-HE group and the severity of the clinical features were significantly higher than those in the CHB group. The former group had a poorer prognosis with a 49.6% occurrence rate of severe hepatitis and a 25.2% death rate while in the latter group the corresponding rates were just 4.4% and 1.7%.
CONCLUSIONRe-infection with HEV, in those CHB patients, severely damaged their hepatic functions and increased their mortality; the older the patient and the higher the level of HBV DNA, the poorer the prognosis.
Female ; Hepatitis B virus ; Hepatitis B, Chronic ; virology ; Hepatitis E ; virology ; Hepatitis E virus ; Humans ; Male ; Middle Aged ; Prognosis ; Superinfection