3.Cloning and functional characterization of a cDNA encoding isopentenyl diphosphate isomerase involved in taxol biosynthesis in Taxus media.
Tian SHEN ; Fei QIU ; Min CHEN ; Xiao-zhong LAN ; Zhi-hua LIAO
Acta Pharmaceutica Sinica 2015;50(5):621-626
Taxol is one of the most potent anti-cancer agents, which is extracted from the plants of Taxus species. Isopentenyl diphosphate isomerase (IPI) catalyzes the reversible transformation between IPP and DMAPP, both of which are the general 5-carbon precursors for taxol biosynthesis. In the present study, a new gene encoding IPI was cloned from Taxus media (namely TmIPI with the GenBank Accession Number KP970677) for the first time. The full-length cDNA of TmIPI was 1 232 bps encoding a polypeptide with 233 amino acids, in which the conserved domain Nudix was found. Bioinformatic analysis indicated that the sequence of TmIPI was highly similar to those of other plant IPI proteins, and the phylogenetic analysis showed that there were two clades of plant IPI proteins, including IPIs of angiosperm plants and IPIs of gymnosperm plants. TmIPI belonged to the clade of gymnosperm plant IPIs, and this was consistent with the fact that Taxus media is a plant species of gymnosperm. Southern blotting analysis demonstrated that there was a gene family of IPI in Taxus media. Finally, functional verification was applied to identify the function of TmIPI. The results showed that biosynthesis of β-carotenoid was enhanced by overexpressing TmIPI in the engineered E. coli strain, and this suggested that TmIPI might be a key gene involved in isoprenoid/terpenoid biosynthesis.
Amino Acid Sequence
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Carbon-Carbon Double Bond Isomerases
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genetics
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Cloning, Molecular
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DNA, Complementary
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genetics
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Escherichia coli
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Paclitaxel
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biosynthesis
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Phylogeny
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Plant Proteins
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genetics
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Taxus
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enzymology
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genetics
4.Detection of soluble interleukin-2 receptor and T lymphocyte subsets in silicosis.
Yu-hua ZHU ; Yue-qiu TIAN ; Guo-ying ZHU ; Hong-zhen ZHANG ; Zhong-xing CAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(1):66-66
Aged
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Humans
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Male
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Middle Aged
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Receptors, Interleukin-2
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blood
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Silicosis
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blood
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classification
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T-Lymphocyte Subsets
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metabolism
5.Inhibition of allogeneic T-cell responses by Kupffer cells expressing indoleamine 2,3-dioxygenase in vitro
Maolin YAN ; Yaodong WANG ; Yifeng TIAN ; Zhide LAI ; Funan QIU ; Songqiang ZHOU ; Shen YOU ; Zhong CHEN
Chinese Journal of Hepatobiliary Surgery 2011;17(3):235-238
Objective To investigate kupffer cells (KCs) expressing indoleamine 2,3-dioxygenase(IDO)in the inhibition of allogeneic T-cell proliferation in vitro. Methods Real-time PCR was used to investigate the expression of IDO mRNA and FasL mRNA in KCs pretreated with or without IFNγ. High performance liquid chromatography was used to analyze the catabolism of tryptophan by IDO from KCs. Allogeneic T-cell response was used to confirm the inhibition of KCs in vitro. The proliferation of lymphocytes was detected using [3 H] thymidine incorporation. Cell cycle and lymphocyte apoptosis were evaluated by flow cytometric assay. Results Real-time PCR revealed IDO mRNA and FasL mRNA expression in KCs pretreated with IFN-γ. IDO catabolic effect was confirmed by a decrease in tryptophan and increase in kynurenine concentration. KCs expressing IDO and FasL from BABL/c mice acquire the ability to suppress the proliferation of T-cells from C57BL/6, which could be blocked by the addition of 1-methyl-tryptophan and anti-FasL antibody. The co-cultured T-cells with KCs expressing IDO and FasL could induce allogeneic T-cell apoptosis and exhibited cell-cycle arrest in G1. Conclusion In addition to the Fas/FasL pathway, IDO may also play an important role in KCs to inhibit allogeneic T-cell proliferation in vitro.
6.Effect of Yiqi Huoxue Qingre Huashi Recipe on the Eradication Rate of Hp in Peptic Ulcer Patients.
Gao-zhong DAI ; Xian-jing FAN ; Qiu-shi TIAN ; Shi-kai ZHU ; Ke-xue ZHAO ; Dan-lei SHEN
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(12):1437-1441
OBJECTIVETo observe the effect of Yiqi Huoxue Qingre Huashi Recipe (YHQHR, a recipe capable of supplementing qi, activating blood, clearing heat, and dissipating dampness) on ulcer healing and Helicobacter pylori (Hp) eradication rate in Hp positive peptic ulcer patients, and to explore coccoid Hp occurrence in the eradication.
METHODSTotally 80 Hp positive peptic ulcer patients were assigned to the treatment group and the control groups by random digit table, 40 in each group. All patients received standard triple therapy of Western medicine for 2 successive weeks. Those in the control group additionally took omeprazole enteric coated tablet, 20 mg each time, once per day for 4 successive weeks. Those in the treatment group additionally took YHQHR, twice per day for 6 successive weeks. The ulcer healing was observed and recorded by gastroscope after discontinued medication of 14 days. The effective rate of ulcer healing under endoscope was statistically calculated. Rapid urease test (RUT) was performed in one small piece of tissue from corpora ventriculi and sinuses ventriculi using 14C breathe test (UBT). Gastric juice was collected from the stomach. Hp urease gene amplification test (urea A-PCR) was performed in living tissue from gastric antrum. Results obtained from the above three test methods were recorded and assessed to decide the final eradiation rate. Gastric mucosa tissue was observed under electron microscope,attempting to find non-eradicated Hp, which was further observed.
RESULTSThe total curative effect under gastroscope was 97.5% (39/40 cases) in the treatment group, obviously higher than that in the control group (80.0%, 32/40 cases) (P < 0.05). The eradication rate of Hp was 75.0% (30/40 cases), obviously better than that of the control group (52.5%, 21/40 cases) (P < 0.05). The total positive Hp numbers after treatment was 14C UBT (12), RUT (8), and urea A-PCR (27), respectively. The Hp positive rate detected by 14C UBT and RUT was lower than the Hp positive rate detected by urea A-PCR (P < 0.05). Rod-like and coccoid Hp bacteria could be observed under electron microscope.
CONCLUSIONYHQHR combined standard triple therapy was more effective than standard triple therapy alone in promoting ulcer healing and elevating the eradication rate of Hp.
Breath Tests ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; Humans ; Omeprazole ; Peptic Ulcer ; drug therapy ; microbiology ; Urea
7.Effect of salvianolic acid B on neural cells damage and neurogenesis after brain ischemia-reperfusion in rats.
Jing ZHONG ; Min-ke TANG ; Yan ZHANG ; Qiu-ping XU ; Jun-tian ZHANG
Acta Pharmaceutica Sinica 2007;42(7):716-721
This study is to observe the effect of salvianolic acid B (Sal B) on neural cells damage and neurogenesis in sub-granular zone (SGZ) and sub-ventricular zone (SVZ) after brain ischemia-reperfusion (I/R) in rats. A modified middle cerebral artery occlusion (MCAO) model of focal cerebral ischemia-reperfusion was used. The rats were divided into four groups: sham control group, ischemia-reperfusion group, Sal B 1 and 10 mg x kg(-1) groups. Sal B was consecutively administrated once a day by ip injection after MCAO. The neurogenesis in SGZ and SVZ was investigated by BrdU method 7 days after MCAO. The Nissl staining for neurons in the hippocampal CA1 and cerebral cortex was performed 14 days after MCAO. A beam-walking test was used to monitor the motor function recovery. We found that brain ischemia resulted in an increase of BrdU positive cells both in ipsilateral SGZ and SVZ at 7th day after MCAO. Sal B (10 mg x kg(-1)) significantly increased further the number of BrdU positive cells both in SGZ and SVZ (P < 0.01). Ipsilateral hippocampal neuron damage occurred and CA1 almost lost 14 days after MCAO. Sal B (10 mg x kg(-1)) obviously attenuated the neuron damage and increased the number of neuron both in ipsilateral CA1 and cerebral cortex (P < 0.01). We also observed an obvious improvement of motor function recovery when Sal B (10 mg x kg(-1)) administrated. From the results above we concluded that Sal B stimulated neurogenesis process both in SGZ and SVZ after brain ischemia, and also alleviated neural cells loss and improved motor function recovery after brain ischemia in rats.
Animals
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Benzofurans
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isolation & purification
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pharmacology
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Cell Count
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Cerebral Cortex
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pathology
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Cerebral Ventricles
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pathology
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Dentate Gyrus
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pathology
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Hippocampus
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pathology
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Infarction, Middle Cerebral Artery
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complications
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Male
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Motor Activity
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drug effects
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Neurogenesis
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drug effects
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Neurons
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drug effects
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pathology
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Plants, Medicinal
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chemistry
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Random Allocation
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Rats
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Rats, Sprague-Dawley
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Reperfusion Injury
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etiology
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pathology
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physiopathology
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Salvia miltiorrhiza
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chemistry
8.Bacteriological Study on the Skin Lesions of Patients with Eczema and Atopic Dermatitis
Zhigang BI ; Meihua TIAN ; Lin LIN ; Ning QIU ; Fei HAO ; Baiyu ZHONG ; Ruofei YIN ; Fanqin ZENG ; Bian ZHAO
Chinese Journal of Dermatology 2003;0(10):-
Objective To study the bacteriological characteristics and the pathogenesis of Staphylococcus aureus (S. aureus) on eczema and atopic dermatitis (AD). Methods A multi-center randomized, double blind bacteriological study on the lesions and non-lesional skin of patients with eczema (207) and AD (119) were carried out. The antibiotic sensitivity and the bacteriophage typing were performed on all the S. aureus isolated from the patients. Results There were statistical differences in the positive rate of the culture, the ratio and the colonization of S. aureus between the lesion and the non-lesional skin in eczema (P
9.Chemical investigation of triterpenoids from Dichrocephala benthamii.
Bo SONG ; Qiu-bo ZHANG ; Meng-hua WANG ; Xin-hui TIAN ; Hao-li SUN ; Fu-bao ZHANG ; Zhong-mei ZOU ; Gang DING
China Journal of Chinese Materia Medica 2015;40(11):2144-2147
The triterpenoids of Dichrocephala benthamii were investigated by means of silica gel, Sephadex LH-20 and semi-preparative HPLC. Nine triterpenoids were isolated from D. benthamii. By analysis of the EI-MS, NMR spectra and comparison to the data reported in literatures, the structures of these compounds were determined as β-amyrin formiate (1), β-amyrin acetate (2), β-amyrenol (3), β-amyrone (4), 3β-hydroxy-olean-11, 13 (18)-diene (5) , Δ12-oleanene (6) , friedelin (7), dammaradienyl acetate (8), epi-friedeband (9), respectively. Compounds 1-8 were isolated for the first time form this genus, compound 9 was isolated for the first time from this plant, whereas β-amyrin formiate (1) was a new natural product.
Asteraceae
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chemistry
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Triterpenes
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chemistry
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isolation & purification
10.Effects of 17β-estradiol and estrogen receptor modulator on the expression of cadherin and catenin in the uterus of ovariectomized rats.
Zhong-Rui DU ; Hong-Fang LI ; Wei ZHANG ; Zhi-Feng TIAN ; Xiao-Qing QIU ; Yan-Yu LU ; Qian-Kun LIANG ; Long-De WANG
Chinese Journal of Applied Physiology 2014;30(5):459-I
Animals
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Cadherins
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metabolism
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Catenins
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metabolism
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Estradiol
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pharmacology
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Estrogen Receptor Modulators
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pharmacology
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Female
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Ovariectomy
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Rats
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Uterus
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drug effects
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metabolism