2.Codon optimization and eukaryotic expression analysis of the analgesic peptide gene BmK AngM1 from Buthus martensii Karsch.
Jinling YANG ; Lili GAO ; Ping ZHU ; Qi HOU ; Fen WANG ; Wenbo YU ; Tao NIE
Acta Pharmaceutica Sinica 2012;47(10):1389-93
Codon bias is an important factor which influences heterologous gene expression. Optimizing codon sequence could improve expression level of heterologous gene. In order to improve the expression level of BmK AngM1 gene encoding the analgesic peptide from Buthus martensii Karsch in Pichia pastoris, the codon-optimized BmK AngM1 gene according to its cDNA sequence and the preference codon usage of P. pastoris were cloned into expression vector pPIC9K and then transformed into P. pastoris. The expersion of recombinant BmK AngM1 (rBmK AngM1) was inducced by methanol in the medium, and the expression level of the optimized BmK AngM1 gene was 3.7 times of the native one. These results suggested that the expression of BmK AngM1 in P. pastoris could be successfully improved by codon optimization.
3.Clinical significance of non - mydriatic fundus photography in screening for preschool children ocular fundus disease
Jun, LUO ; Yi-Lan, TAN ; Fen, DU ; Yu, CHEN ; Li-Juan, TAO
International Eye Science 2014;(6):1179-1180
AIM:To observe the incidence of ocular fundus disease in preschool children examined by non-mydriatic fundus camera and evaluate its effectiveness compared with direct inspection shadow mirror.
METHODS: Three thousand eight hundred and ninety-six preschool children from April 2012 to October 2013 were examined by Topcon TRC-NW300 color fluorescence fundus camera and direct inspection shadow mirror, and images were saved immediately.
RESULTS: Detection rate of non - mydriatic fundus photography was higher than that of direct inspection shadow mirror. In 3 896 cases, 41 eyes were detected abnormal fundus accounting for 1. 05%. The retinal myelinated nerve fibers, morning glory syndrome, retinitis pigmentosa, congenital retinoschisis were common, accounted for 24. 39%, 21. 95%, 14. 63%, 12-20% respectively. The children eye diseases were often accompanied by abnormal vision (68. 30%), ametropia (63. 41%), strabismus (19. 51%).
CONCLUSION:Non-mydriatic fundus photography is a mydriatic method without medicine, so it is easy for preschool children to accept. Image results could directly display the fundus lesions. It shows important significance in the screening for preschool children eye diseases.
4.Protective mechanism of Danhong injection on brain microvascular endothelial cells injured by hypoxic.
Peng ZHOU ; Yu HE ; Jie-hong YANG ; Yu-yan ZHANG ; Hui-fen ZHOU ; Tao ZHAO ; Wei FU ; Hai-tong WAN
China Journal of Chinese Materia Medica 2014;39(24):4844-4848
To study the protective mechanism of Danhong injection on brain microvascular endothelial cells (rBMECs) injured by hypoxic. In the experiment, primary suckling mouse's rBMECs cells were collected and identified with factor VIII to establish the 4 h injury model. Meanwhile, rBMECs were given Danhong injection (25, 50, 100 mL . L-1), and the superoxide dismutase (SOD) activity and the malonyldialdehyde (MDA) level were detected by the biochemical method. Cell MMP-9, ICAM-1 and P53 mRNA expression levels were detected by RT-PCR method. Changes in cells' microscopic structure were observed by transmission electron microscope. According to the results, primary rBMECs were notably injured by hypoxia. Compared with model group, Danhong injection (50, 100 mL . L-1) could remarkably resist the injury induced by hypoxic, increase intracellular SOD activity, decrease MDA level and significantly down-regulate ICAM-1, MMP-9 and P53 mRNA expressions. Danhong injection (100 mL . L-1) could protect the cells' normal morphology and microscopic structure, maintain the close intercellular junction, and inhibit the hypoxia-induced cell apoptosis. The results showed that Danhong injection plays a significant role in protecting rBMECs injured by hypoxia. Its mechanism may be related to the enhancement of cells' antioxidant capacity, the inhibition of inflammatory response and the cell apoptosis.
Animals
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Antioxidants
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metabolism
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Apoptosis
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drug effects
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Brain
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metabolism
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Cell Hypoxia
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Endothelial Cells
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ultrastructure
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Female
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Gene Expression Regulation
;
drug effects
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Humans
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Injections
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Intercellular Adhesion Molecule-1
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metabolism
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Male
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Malondialdehyde
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metabolism
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Matrix Metalloproteinase 9
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genetics
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Protective Agents
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pharmacology
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Rats
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Rats, Sprague-Dawley
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Superoxide Dismutase
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metabolism
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Tumor Suppressor Protein p53
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genetics
5.Protective effect of combined administration of active ingredients of Danhong on cerebral micro-vascular endothelial cell injured by hypoxia.
Hui-fen ZHOU ; Yu HE ; Yu-yan ZHANG ; Jie-hong YANG ; Tao ZHAO ; Wei FU ; Peng ZHOU ; Hai-tong WAN
China Journal of Chinese Materia Medica 2014;39(22):4399-4404
To study the protective effect of combined administration of active ingredients of Danhong on cultured primary mice's brain microvascular endothelial cells (rBMECs) injured by hypoxia. Primary mice's brain micro-vascular endothelial cells were cultured to establish the 4 h hypoxia model. Meanwhile, active ingredients (protocatechuic aldehyde, salvianolic acid B, hydroxysafflor yellow A and tanshinol) of Danhong were administered in rBMECs. The non-toxic dosage was determined by MTT. The leakage of lactate dehydrogenase(LDH), cell superoxide dismutase (SOD) activity and MDA level were detected by the colorimetric method. The expressions of ICAM-1, MMP-9, P53 mRNA were detected by RT-PCR method. Changes in rBMECs cell cycle and early apoptosis were detected by flow cytometry. Danhong's active ingredients and prescriptions 1, 2, 3, 7, 8, 9 could be combined to significantly restrain LDH in hypoxic cells supernatant. Prescriptions 1, 2, 3, 7, 8, 9 could significantly enhance SOD activity in anoxic cells; Prescriptions 1, 2, 3, 8, 9 could significantly decrease the MDA level; Prescriptions 1, 2, 6, 7, 9 could significantly inhibit the early rB-MECs apoptosis induced by hypoxia. After hypoxia, the up-regulated P53 mRNA expression could cause retardation in G, phase and promote cell apoptosis. This proved that the regulatory function of P53 gene lay in monitoring of calibration points in G, phase. Prescriptions 1, 2, 5, 6, 7, 8, 9 could significantly down-regulate the P53 mRNA expression; Prescriptions 1, 4, 7, 8, 9 could significantly down-regulate the ICAM-1 mRNA expression; Prescriptions 1, 3, 6, 9 could significantly down-regulate the MMP-9 mRNA expression. The combined administration of Danhong's active ingredients showed a significant protective effect on primary cultured rBMECs injury induced by hypoxia Its mechanism may be related to the enhancement of cellular antioxidant capacity and the inhibition of inflammatory response and cell apoptosis. This study could provide ideas for researching prescription compatibility, and guide the clinical medication.
Animals
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Apoptosis
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drug effects
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Brain
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drug effects
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Drugs, Chinese Herbal
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chemistry
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pharmacology
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Endothelial Cells
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drug effects
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Hypoxia
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drug therapy
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Microvessels
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drug effects
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Rats
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Rats, Sprague-Dawley
6.Studies on terpenoids from Zygophyllum fabago.
Jiang-ho HE ; Yan-fen NIU ; Jin-xian LI ; Lin-bo WANG ; Tai-ping ZI ; Shan YU ; Jian TAO
China Journal of Chinese Materia Medica 2015;40(23):4634-4638
This study was to investigate the chemical constituents of the aerial part of Zygophyllumfabago, by phytochemical methods. The compounds were isolated by silica gel and Sephadex LH-20 column chromatographies from the EtOAc extract. Their structures were characterized by various spectroscopic data (1H-NMR, 13C-NMR, MS) and comparison with the literature. As a result, thirteen compounds were isolated and their structures were identified as 1-hydroxyhinesol(1), hinesol(2), atractylenolactam(3), beta-eudesmol (4), 5alpha-hydroperoxy-beta-eudesmol(5), 12-hydroxy-valenc-1(10)-en-2-one(6), pubinernoid A(7), (6S,7E)-6-hydroxy-4,7-megastigmadien-3,9-dione(8), 3-hydroxy-5alpha, 6alpha-epoxy-beta-ionone (9), (3S,5R, 6S, 7E)-3, 5, 6-trihydroxy-7-megastigmen-9-one(10), (6R,7E,9R)-9-hydroxy-4,7-megastigmadien-3-one(11), (S)-3-hydroxy-beta-ionone(12), and blumenol A(13). Compounds 1-7 were sesquiterpenoids and 8-13 were megastigmane type norsesquiterpenoids. All the compounds were obtained from Z. fabago for the first time, and compound 1 was a new natural product.
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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Terpenes
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chemistry
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isolation & purification
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Zygophyllum
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chemistry
7.Cytology diagnosis and clinical management of fine-needle aspiration for thyroid nodules.
Jin-yu ZHENG ; Tao BAI ; Yi-fen ZHANG ; A-qing CHEN ; Qin HUANG
Chinese Journal of Pathology 2010;39(5):349-352
Biopsy, Fine-Needle
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Carcinoma
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diagnosis
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pathology
;
surgery
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Carcinoma, Medullary
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diagnosis
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pathology
;
surgery
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Carcinoma, Papillary
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diagnosis
;
pathology
;
surgery
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Carcinoma, Papillary, Follicular
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diagnosis
;
pathology
;
surgery
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Diagnosis, Differential
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Goiter, Nodular
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diagnosis
;
pathology
;
therapy
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Hashimoto Disease
;
diagnosis
;
pathology
;
therapy
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Humans
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Lymphoma
;
diagnosis
;
pathology
;
surgery
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Thyroid Nodule
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pathology
;
surgery
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Thyroidectomy
8.Effect and mechanism of Guhong injection against cerebral ischemia reperfusion injury.
Ming-chun SHU ; Hai-tong WAN ; Hui-fen ZHOU ; Jie-hong YANG ; Tao ZHAO ; Wei FU ; Yu HE
China Journal of Chinese Materia Medica 2014;39(24):4829-4833
To explore the effect and mechanism of Guhong injection against cerebral ischemia reperfusion injury, SD rats were randomly divided into the sham-operated group, the model group, the nimodipine group, and high, medium and low-dose Guhong injection groups, with 10 rats in each group. The middle cerebral artery embolization (MCAO) model was established to observe neurological deficit symptoms, infarct volume, SOD activity, MDA content, GSH-Px and CAT activity in rats, as well as the contents of t-PA, PAI, TXB2 and 6-keto-PGF1α in serum. The results showed that Guhong injection could obviously promote the recovery of neurological deficit symptoms, narrow the brain infarct volume in rats after surgery, significantlyincrease the activities of SOD, GSH-Px and CAT and decrease the content of MDA. Meanwhile, it also could obviously increase the contents of t-PA and 6-keto-PGF1α and decrease the contents of PAI and TXB2 in serum, indicating that Guhong injection have better antioxidant and antithrombus effects, as well as a significant protective effect against cerebral ischemia reperfusion injury in rats.
Animals
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Antioxidants
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pharmacology
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Brain Ischemia
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drug therapy
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Catalase
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metabolism
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Drugs, Chinese Herbal
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pharmacology
;
Injections
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Male
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Malondialdehyde
;
metabolism
;
Random Allocation
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Rats
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Rats, Sprague-Dawley
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Reperfusion Injury
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drug therapy
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Superoxide Dismutase
;
metabolism
9.Protective effect of Shenxiong injection on cerebral ischemia-reperfusion injury of rats.
Xing-Yu WANG ; Yu-Yan ZHANG ; Hai-Tong WAN ; Tao ZHAO ; Wei FU ; Hui-Fen ZHOU ; Peng ZHOU ; Ze-Feng WANG ; Jie-Hong YANG
China Journal of Chinese Materia Medica 2014;39(3):503-506
OBJECTIVETo study the protective effect of Shenxiong injection on the cerebral ischemia-reperfusion injury of senile rats.
METHODTotally 108 Sprague-Dawley (SD) rats were randomly divided into the sham operation group, the model group, the Ni-modipine group and Shenxiong injection groups (low, middle, and high doses). The rat brain ischemia-reperfusion model was established by the middle cerebral artery occlusion (MCAO) method in rats, in order to observe the effect of Shenxiong injection on neurological score and brain infarct volume of rats with cerebral ischemia-reperfusion injury, and determine the contents of NOS, NO, SOD, MDA and LDH in brain tissues. The contents of TNF-alpha and IL-1beta levels in brain tissues were measured by enzyme-linked immunosorbent assay (ELISA) method.
RESULTShenxiong injection could significantly decrease neurological score, injury degree of brain tissues and brain infarct volume of rats with cerebral ischemia-reperfusion injury, increase the vigor of SOD, decrease the levels of MDA, NO, NOS and LDH, and inhibit IL-1beta and TNF-alpha expressions.
CONCLUSIONShenxiong injection has the obvious protective effect on the brain ischemia-reperfusion injury in rats. Its mechanism may be related to the improvement of neurological function, the reduction of free radical injury, and the inhibition of inflammation factor expression.
Animals ; Brain ; blood supply ; drug effects ; metabolism ; Brain Ischemia ; complications ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Injections ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; drug therapy ; enzymology ; metabolism ; Superoxide Dismutase ; metabolism
10.Effect of Shenxiong injection on inflammation injury of ischemia-reperfusion injury rats.
Hui-fen ZHOU ; Jin-chao AL ; Hai-tong WAN ; Yu HE ; Yu-yan ZHANG ; Tao ZHAO ; Wei FU ; Jie-hong YANG
China Journal of Chinese Materia Medica 2015;40(12):2408-2412
To investigate the effect of Shenxiong injection on the inflammation injury of ischemia-reperfusion injury senile rats. Totally 84 Sprague-Dawley (SD) rats were randomly divided into six groups: the sham operation group, the model group, the Nimodipine group and the Shenxiong injection(low, middle, and high dosage) groups. The rat cerebral ischemia-reperfusion model was established through intraperitoneal injection for 3 d and middle cerebral artery occlusion (MCAO). Ater the reperfusion for 24 h, efforts were made to give neurological score, collect brains for TTC staining, detect tumor necrosis factor-α(TNF-α) and interleukin-1β(IL-1β) content in serum by enzyme-linked immunosorbent assay (ELISA) method and measure IL-1β, ICAM-1 and MMP-9 mRNA expressions in hippocampal area by Real-time PCR (RT-PCR). According to the results, Shenxiong injection could decrease the cerebral infarction volume, greatly improved the neurological function and reduce IL-1β, TNF-α, ICAM-1 and MMP-9 mRNA expressions and IL-1β and TNF-α contents. In conclusion, Shenxiong injection shows the significant protective effect on ischemia-reperfusion injury in rats. Its mechanism may be related to the inhibition of inflammatory factor expression.
Animals
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Drugs, Chinese Herbal
;
administration & dosage
;
Humans
;
Intercellular Adhesion Molecule-1
;
genetics
;
immunology
;
Interferon-alpha
;
genetics
;
immunology
;
Interleukin-1beta
;
genetics
;
immunology
;
Male
;
Matrix Metalloproteinase 9
;
genetics
;
immunology
;
Rats
;
Rats, Sprague-Dawley
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Reperfusion Injury
;
drug therapy
;
genetics
;
immunology