3.Expression of recombinant human acetylcholinesterase and its application in screening its inhibitors.
Xiang-Jun WANG ; Huai-Xiu WU ; Shan-Shan YE ; Lan-Ying PAN ; Yong-Chang QIAN
Acta Pharmaceutica Sinica 2014;49(1):50-54
This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.
Acetylcholinesterase
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genetics
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metabolism
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Cholinesterase Inhibitors
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analysis
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pharmacology
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HEK293 Cells
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Humans
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Indans
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analysis
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pharmacology
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Inhibitory Concentration 50
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Kinetics
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Piperidines
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analysis
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pharmacology
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Plasmids
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Recombinant Proteins
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genetics
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metabolism
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Transfection
4.Experimental study on relationship between TCM syndrome type and gastric mucosa cell proliferation in rat model of chronic atrophic gastritis.
Shan XU ; Chang-song WANG ; Jia-he ZHOU
Chinese Journal of Integrated Traditional and Western Medicine 2007;27(11):1004-1006
OBJECTIVETo explore the relationship between TCM syndrome type and gastric mucosa cell proliferation related controlling gene protein in chronic atrophic gastritis (CAG).
METHODSExpressions of cell proliferation related controlling gene protein, including proliferative cell nuclear antigen (PCNA), epidermal growth factor receptor (EGFR) and c-myc, from gastric mucosa of CAG model rats with different syndrome types were measured by immunohistochemistry and the changes of them before and after TCM intervention were also analyzed by image analysis.
RESULTSProtein expressions of PCNA, EGFR and c-myc in gastric mucosa of CAG model rats with different syndrome types (Pi-deficiency type, Gan-stagnation type and dampness-heat type) were different to some extent, and all of them reduced significantly after TCM intervention in the model rats of all syndrome types.
CONCLUSIONExpressions of cell proliferation related controlling gene protein in gastric mucosa of CAG model rats of different syndrome types were different to some extent, which provides a certain experimental evidence for revealing the essence of TCM syndrome type of CAG and judging the prognosis of various types.
Animals ; Diagnosis, Differential ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; drug effects ; metabolism ; pathology ; Gastritis, Atrophic ; diagnosis ; drug therapy ; metabolism ; Immunohistochemistry ; Male ; Medicine, Chinese Traditional ; Phytotherapy ; Proliferating Cell Nuclear Antigen ; biosynthesis ; Proto-Oncogene Proteins c-myc ; biosynthesis ; Random Allocation ; Rats ; Rats, Wistar ; Receptor, Epidermal Growth Factor ; biosynthesis ; Syndrome
5.Study on winter dormancy of Thesium chinense and its phenological phase.
Ling-shan SONG ; Xiao-ming ZHANG ; Qiao-sheng GUO ; Lu CHEN ; Chang-lin WANG
China Journal of Chinese Materia Medica 2015;40(23):4585-4590
In order to explore reasonable artificial cultivation pattern of Thesium chinense, the biological characteristics and nutrients change in the process of winter dormancy of T. chinense was studied. The phenological period of T. chinense was observed by using fixed-point notation and the starch grains changes were determined dynamically by PAS-vanadium iron hematoxylin staixjing method. Soluble sugar and starch content were measured by anthrone-sulfuric acid method and amylase activity was determined by DN'S method. The results showed that the normal life cycle of T. chinense was two years. T. chinense was growing by seed in the first year, but growing by the root neck bud in the second year. During the process of dormancy, starch and soluble sugar could mutual transformation in different periods. T. chinense had sufficient carbohydrate to maintain growth and also a lot of small molecules to improve their ability to fight against adversity.
Plant Dormancy
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Plant Leaves
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chemistry
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growth & development
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metabolism
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Plant Roots
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chemistry
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growth & development
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metabolism
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Plant Stems
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chemistry
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growth & development
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metabolism
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Santalaceae
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chemistry
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growth & development
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metabolism
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Seasons
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Starch
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analysis
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metabolism
7.Research on the effect of statins on insulin secretion from pancreatic islet in rats and its mechanisms
Baocheng CHANG ; Miaoyan ZHENG ; Chunyan SHAN ; Juhong YANG ; Ying WANG ; Huizhu REN ; Liming CHEN ; Peihua FANG
Chinese Journal of Internal Medicine 2011;50(5):393-396
Objective To evaluate the inhibitory effect of statins on glucose-stimulated insulin secretion (GSIS) of pancreatic islet in rat and to explore its mechanisms. Methods According to the average volume, freshly isolated or 24-hour cultured pancreatic islets were randomly divided into control group( incubated with Kreb-Ringer bicarbonate buffer), the atorvastatin group( incubated with 100 μ mol/L atorvastatin), the fluvastatin group (incubated with 100 μ mol/L fluvastatin)and the pravastatin group (incubated with 100 μ mol/L pravastatin). Stimulated by 2. 8,5. 5,11.1,16. 7 mmol/L and 25.0 mmol/L glucose respectively, the effect of 100 μ mol/L statins on ATP content and GSIS was compared in the four groups. GSIS was performed by the 37℃ bath incubation method and ATP content was measured by chemiluminescence method. Results Incubated with 100 μ mol/L atorvastatin for 30 minutes, in the presence of 16. 7 mmol/L glucose, the ATP content [(9. 54 ± 1. 64) pmol/islet vs ( 12. 33 ± 1.89) pmol/islet] and GSIS (1.60 ± 0. 21 vs 2. 39 ± 0. 30) were significantly reduced in comparison with the control group (P<0. 05). Cultured with 100 μmol/L fluvastatin for 24 hours, the ATP content [( 10. 24 ±2.01 )pmol/islet vs (12. 31 ±2. 16) pmol/islet] and GSIS (3. 12 ± 0. 32 vs 4. 17 ±0. 37 ) were all significantly decreased at the higher glucose concentration of 16. 7 mmol/L ( P < 0. 05). Conclusion Atorvastatin and fluvastatin may inhibit GSIS by decreasing ATP content in pancreatic islet and the inhibitory effect is related to the strength of its lipophilicity.
8.Efficacy of nateglinide combined with metformin in patients with newly diagnosed drug-na(i)ve type 2 diabetes
Baocheng CHANG ; Miaoyan ZHENG ; Chunyan SHAN ; Hechao LI ; Xinli WANG ; Jianxin ZHOU ; Liming CHEN
Chinese Journal of General Practitioners 2011;10(2):93-96
Objective To compare efficacy of nateglinide or acarbose combined with metformin in patients with newly diagnosed type 2 diabetes.Methods Ninety-six patients with newly diagnosed type 2 diabetes in Metabolic Diseases Hospital of Tianjin Medical University,were randomly to receive nateglinide combined with metformin (group A,n =46) or acarbose combined with metformin (group B,n =42) for four months.Drug dose was adjusted every two weeks.Before and after treatment,oral glucose tolerance test and insulin release test were performed to observe changes of their glucose tolerance,homeostasis model assessment for insulin resistance (HOMA-IR) ,insulin secretion function of β-cells and glucose disposition index (DI) in the two groups.Results After four-month treatment,six patients restored to normal glucose tolerance and 13 patients returned to impaired glucose tolerance (IGT) in group A and 12 patients restored to IGT in group B.Their HOMA-IR was markedly improved compared with baseline in both groups,decreased to 7.1 ± 1.3 from 8.6 ± 1.2 in group A and to 6.9 ± 1.7 from 8.6 ± 1.7 in group B ( P < 0.05 ).Compared with group B,early insulin secretion ( LN△I30/△G30 ) obviously improved in group A [( 1.9 ±0.8) vs.(1.6±0.6) mU/mmol] (P<0.05) and DI also increased (1.05±0.25 vs.0.89±0.21,P<0.05 ).Conclusions Nateglinide combined with metformin can obviously restore early insulin secretion and improve insulin resistance in patients with newly diagnosed type 2 diabetes,thus facilitate restoration of their glucose tolerance.
9.Development of China Musculoskeletal Questionnaire and item selection.
Shan-shan WU ; Li-hua HE ; Sheng WANG ; Jian-xin WANG ; Jing-yun LI ; Lei CAO ; Wei-wei DU ; Lei YANG ; Shan-fa YU ; Yan ZHANG ; Chang-yun ZHANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2011;29(12):908-912
OBJECTIVETo develop a draft questionnaire (China Musculoskeletal Questionnaire, CMQ) for evaluating of musculoskeletal workload and associated potential hazardous working conditions as well as musculoskeletal symptoms of workers in Sitting Posture.
METHODSMulti-methods, which include the reviewing references, the summarizing results of preliminary studies, the reviewing ergonomic tools, the consulting experts and occupational health workers and the interviewing or discussing with individual workers in sitting posture, were used in developing item pool. The experts and epidemiologists of occupational health scored the importance of every single item in the item pool, and then the survey and sampling were carried out in 325 workers of sitting posture who completed the questionnaire. On the basis of these data, the methods including experts scoring, item analysis, Cronbach's α analysis and factor analysis were synthetically used to select the reliable items which consisted of the formal questionnaire.
RESULTSThe standard of the CMQ, which consists of 34 items on musculoskeletal workload and associated potentially hazardous working conditions, can be divided into nine indices (dynamic loads, static loads, repetitive loads, forces-exertion, prolong time, climatic factors, vibration, position and ergonomic environmental factors).
CONCLUSIONThe CMQ possesses good content validity, and the items of CMQ are divergent, reliable and typical. However, the reliability and validity of CMQ should be validated.
China ; Ergonomics ; Humans ; Musculoskeletal System ; Occupational Health ; Posture ; Surveys and Questionnaires ; Workload
10.Effect of Ganoderma lucidum polysaccharides on oxidative stress of hyperlipidemic fatty liver in rats.
Shan-Shan CHANG ; Dan ZHOU ; Guo-Liang MENG ; Feng WU ; Shuang WANG ; Xiao CHEN ; Ji-Liang XU
China Journal of Chinese Materia Medica 2012;37(20):3102-3106
OBJECTIVETo study the effect of Ganoderma lucidum polysaccharides on oxidative stress of hyperlipidemic fatty liver in rats.
METHODSeventy-two SD rats were randomly divided into six groups, namely the normal control group (NG), the model group (MG), the G. lucidum polysaccharides groups of low, middle and high dose (GLPs-LG, GLPs-MG, GLPs-HG) and the Simvastatin group (SV). The rats were fed with high fat diet to establish the model of hyperlipidemic fatty liver in rats. After administration for 12 weeks, rats in each group were tested with the following indexes: total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and total antioxidant capacity (T-AOC) in serum as well as the contents of SOD, MDA, GSH-Px and T-AOC in hepatic tissues. Histopathological changes of hepatic tissues were observed under light glass.
RESULTSThe contents of TC, TG and LDL-C were significantly increased in the model group (P < 0.01). Compared with the model group, both the GLPs-M group and the GLPs-H group showed significant decreases in TC, TG and LDL-C (P < 0.05 or P < 0.01), while the GLPs-H group showed a notable increase in HDL-C (P < 0.05). Compared with the model group, both the GLPs-M group and the GLPs-H group showed significant decreases in MDA (P < 0.05 or P < 0.01) and notable increases in SOD, GSH-Px, T-AOC (P < 0.05 or P < 0.01). The GLPs-M group and the GLPs-H group proved a remarkable alleviation in fatty degeneration of hepatic cells.
CONCLUSIONG. lucidum polysaccharides can significantly reduce the blood fat level of hyperlipidemic fatty liver in rats and effectively inhibit oxidant stress, showing the effect on preventing and treating hyperlipidemic fatty liver in rats to some extent.
Animals ; Antioxidants ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Fatty Liver ; drug therapy ; metabolism ; Glutathione Peroxidase ; metabolism ; Humans ; Male ; Malondialdehyde ; blood ; Oxidative Stress ; drug effects ; Polysaccharides ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Reishi ; chemistry ; Triglycerides ; blood