1.Changes of memory and M receptor injection of A?_(25-35) into basal ganglion region of forebrain and effect of catalpol
shuo, LIU ; ya-er, HU ; zong-qin, XIA
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(08):-
Objective To establish mouse model of dementia by intracranial injection of A?25-35 and small amount of ibotenic acid(IBO) and to explore whether the effects of catalpol can affect the brain M receptor density and the short term memory. Methods The mice were randomly divided into three groups: control group,model group,treat group which were given orally for 2 months with 50 mg?kg-1?d-1 of catalpol.Dementia model was developed by single unilateral injection of 0.3 ?L of a solution of A?(1?L normal saline containing 4 ?g of A?25-35 and 1 ?g of ibotenic acid) into right basal ganglion region according the atlas of mouse brain with the aid of a stereotaxic equipment.The track of injection was observed by HE staining.The learning/memory ability was measured by Y-maze perfor-mance.The brain muscarinic receptor density was analyzed with single-site binding assay using 3H-quinuclidinyl benzilae(QNB).Results Two months after model development,the learning ability as well as the density of muscarinic receptor in brain were significantly decreased in model mice compared with those in control mice.Parallel models treated with daily oral administration of Catalpol for two months improved the learning ability and increased the brain muscarinic receptor density when compared with model mice.The correlation coefficient between total M receptor densities and the learning/memory ability was significant when examined with linear regresion.Conclusion A dementia model was established in mice.Dementia model was developed by single unilateral injection of 0.3 ?L of a solution of A?(1 ?L normal saline containing 4 ?g of A?25-35 and 1 ?g of ibotenic acid) into right basal ganglion region was established in mice.Catalpol can significantly improve the learning and increase the brain muscarinic receptor density of the model.
2.Production and Preliminary Application of Monoclonal Antibodies Raised Against Aβ22-35
Jing-wei, SHENG ; Ya-er, HU ; Zong-qin, XIA
Journal of Shanghai Jiaotong University(Medical Science) 2001;21(2):110-112,140
Objective To produce a hybridoma secreting stable monoclonal antibodies against Aβ22-35 and to develop a detection method for the assay of Aβ. Methods Spleen cells from Balb/cmice immunized with Aβ22-35-KLH were fused with mouse myeloma cells SP2/0. The techniques of immunoprecipitation and western blotting plus ECL were used to investigate the levels of Aβ in the rat brain. Results Two strains of hybridomas (3A8 and 3B2) secreting stable monoclonal antibodies raised against Aβ22-35 were obtained. The subtypes of Aβ22-35 were IgG3. The levels of Aβ in young and older rat brain were 9.8±2.8 and 13.36±2.65 (pmol/12mg brain tissues, x±s), respectively. Conclusion The Aβ22-35 mAb obtained had high titres and specificity. The levels of Aβ in the older rat brain were significantly increased as compared with the young one (P<0.05).
3.Effects of Cichorium glandulosum Boiss. et Huet. on expression of fibronectin, Smad3, IGFBP-rPl, and TGFβ1 in a liver fibrosis rat model.
Dong-mei QIN ; Li-ping HU ; Ya-ru NIE ; Wen CHEN
Chinese Journal of Hepatology 2013;21(10):776-777
Animals
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Chicory
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Fibronectins
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metabolism
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Insulin-Like Growth Factor Binding Proteins
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metabolism
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Liver
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drug effects
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metabolism
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Liver Cirrhosis, Experimental
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metabolism
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Male
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Plant Extracts
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pharmacology
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Rats
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Rats, Wistar
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Smad3 Protein
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metabolism
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Transforming Growth Factor beta1
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metabolism
4.Effect of catalpol from Radix Rehmanniae on A?_(25-35)-induced apoptosis of PC12 cells
jin-hong, WANG ; rui, ZHANG ; ya-er, HU ; zong-qin, XIA
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(07):-
Objective To investigate the effect of catalpol from Radix Rehmanniae on A?25-35-induced apoptosis of PC12 cells.Methods PC12 cells were routinely cultivated and treated by A?25-35(final concentration,20 ?mol/L) 24 hours after the addition of catalpol or saline.Forty-eight hours later,cells were examined for viability and apoptosis by MTT method and TUNEL method,respectively,while Bax and Bcl-2 mRNA expression were analyzed by semi-quantitive RT-PCR. Results Catalpol could significantly elevate the viability at 1?10-5 mol/L and 1?10-4 mol/L(P
5.Dual direction regulatory effect of catalpol on ?-adrenergic receptor and M-cholinergic receptor
qing-feng, LIU ; zong-qin, XIA ; qi-xiang, SUN ; ya-er, HU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(12):-
Objective To investigate whether catalpol is the active component responsible for the Yin tonic effect of Radix Rehmanniae.Methods Young NH mice were injected with triiodothyronine to produce the hyperthyroidism model,while old mice were used as the model of natural aging.The single point radioligand binding assay was carried out to determine the ?-adrenergic receptor density and M-cholinergic receptor density.The learning ability(short term memory) was determined by the Y-maze avoidance test. Results In the ?-adrenergic receptor experiment,the densities were(15.7?5.2) and(20.9?7.2) fmol/mg protein in normal control group and in T3 control group(P
6.Effects of active component of Zhimu on APP and BACE1 in HEK293sw cells
rui, ZHANG ; zi-mei, WANG ; zong-qin, XIA ; ya-er, HU
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(07):-
Objective To investigate the effects of ZMS,the active component of Zhimu,on amyloid precursor protein(APP) and ?-site APP cleaving enzyme 1(BACE1) in HEK293sw cells. Methods HEK293sw cells were pretreated with different concentrations of ZMS in routine culture medium for 24 h,and then serum-free medium with ZMS was used for further culture for 48 h.Cells were examined for expression of APP and ?-secretase BACE1 with Western blotting and RT-PCR,and were then examined for BACE1 activity with fluorescence method. Results 1 ?mol/L and 10 ?mol/L ZMS significantly decreased the expression of BACE1 mRNA and protein,while had no effect on the expression of APP mRNA and protein.It was indicated by fluorescence analysis that 10 ?mol/L ZMS significantly decreased the ?-secretase activity.Conclusion ZMS significantly decreases the activity and expression of ?-secretase BACE1,while has no effect on the expression of APP in HEK293sw cells.
7.Quality of life assessment and the related factors of patients with active ankylosing spondylitis
Xiao-Hu DENG ; Feng HUANG ; Hui-Qin HAO ; Li-Sha WANG ; Ya-Mei ZHANG ;
Chinese Journal of Rheumatology 2003;0(08):-
Objective To analysis the quality of life of patients with active ankylosing spondylitis (AS).Methods The quality of life was assessed in 52 patients with active AS using SF-36 and was compared with the general population.The correlation between the quality of life and the clinical measures of disease,in- cluding the Bath AS disease activity index(BASDAI),Bath AS functional index(BASFI),Bath AS metrology index(BASMI),patient's global assessment(PGA),spinal inflammation,total back pain,nocturnal back pain, and enthesis index(EI),were determined.Results The patients with active AS reported significant decreased scores on all dimensions of SF-36.The score of physical health components was poorer than that of mental health components.BASFI was the strongest factor correlated with the score of SF-36,followed by BASDAI and PGA.BASMI and total back pain were correlated with three dimensions only.In multivariate regression analysis,BASFI showed relative closer relationship to the qulity of life with active AS than other clinical mea- sures of disease and it accounted for 50.3%,35.2% and 47.9% of the variance in the physical health compo- nents,the mental health components and the overall score of SF-36,respectively.Conclusion The quality of life in patients with active AS is significantly declined compared with general population.The physical aspects seem to be more severely affected.Functional status of the patients with active AS are correlated with the quality of life closely.
8.Physical and chemical characteristics of a new cefazolin sodium hydrate crystal.
Chang-Qin HU ; Li-Hui YIN ; Ya-Ning LIANG
Acta Pharmaceutica Sinica 2008;43(8):868-872
One kind of new cefazolin sodium hydrate crystal was obtained in the isopropyl alcohol - water system. There are two symmetry independent molecules in the asymmetric unit, both being well ordered in the lattice, and ten independent water positions but generally four to six (mean five) water molecules and one sodium ion present in the unit cell structure. Huge solvent tunnels can be found. Again there are two general regions of water molecules, those in the large solvent tunnels and those in proximity of the sodium ion and the tetrazole moieties of the drug molecule. The physical and chemical characteristics of the new cefazolin sodium hydrate crystal are similar to that of the alpha-form cefazolin sodium crystal, and the new crystal has better chemical stability than amorphous cefazolin sodium powder.
Anti-Bacterial Agents
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chemistry
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Cefazolin
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chemistry
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Crystallization
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Crystallography, X-Ray
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Drug Stability
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Molecular Conformation
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Molecular Structure
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Sodium
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analysis
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Spectrophotometry, Atomic
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Water
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chemistry
9.Regulatory Effect of Qushi Huayu Recipe on Gene Expression Profiles of Fatty Liver Rats.
Qin FENG ; Ya-jun TANG ; Xiao-fei LI ; Sheng-xi MENG ; Jing-hua PENG ; Yu ZHAO ; Yi-yang HU
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(2):203-209
OBJECTIVETo observe the intervention and mechanism of Qushi Huayu Recipe (QHR) on gene expression profiles in high lipid diet induced fatty liver rats.
METHODSFatty liver model was prepared in 20 male SD rats using single high fat diet (88% common forage +2% cholesterol +10% lard). Four weeks after modeling they were divided into the model group and the QHR group according to random digit table, 10 in each group. QHR (at 0. 93 g crude drug/100 g body weight) and distilled water was respectively to rats in the QHR group and the model group by gastrogavage while modeling, once per day. Meanwhile, 10 SD male rats were recruited in a normal group, administered with equal volume of distilled water by gastrogavage. At the end of week 8 all rats were sacrificed, and blood and livers were collected for subsequent analysis. Contents of liver triglyceride (TG) and free fatty acid (FFA) , activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected using biochemical assay. Pathological changes of liver tissue were observed using H&E and oil red O stain. Liver gene expressions were detected by Affymetrix gene expression profiles. Differentially expressed genes were compared between the QHR group and the model group, functions of differentially expressed genes and signal pathways involved analyzed. Ten differentially expressed genes involved in glycolipid metabolism with fold change more than 2 were selected for verification by real-time PCR.
RESULTS(1) Compared with the normal group, contents of liver TG and FFA, and serum activities of ALT and AST obviously increased in the model group (P <0. 01). Compared with the model group, contents of liver TG and FFA, and activities of ALT and AST obviously decreased in the QHR group (P <0. 05, P <0. 01). QHR could reduce high fat induced fatty degeneration of liver cells , alleviate inflammation, and improve pathological changes of liver tissue. (2) Compared with the model group, there were 80 differentially expressed genes (with fold change > 2, P < 0.05) with clear functions and appointed gene names, including 44 up-regulated and 36 down-regulated genes. Eighty genes were involved in 27 signal pathways with statistical difference, including glycerolipid metabolism, adipocytokine signaling pathway, insulin signal pathway, drug metabolism signal pathway, etc (P < 0.05). (3) RT-PCR results of 10 glycolipids metabolism regulating genes such as Gk, Scd1, Gpat2, G6pc, Irs1, and so on showed that all RT-PCR genes were completely coincide with up-regulated or down-regulated tendency in results of gene chips. 80% genes had approximate fold change.
CONCLUSIONQHR could regulate gene expressions related to fat metabolism, carbohydrate metabolism, anti-lipid peroxidation, and drug metabolism in high fat diet induced fatty liver rats, and its comprehensive pharmacological actions could be manifested.
Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Carbohydrate Metabolism ; Diet, High-Fat ; Drugs, Chinese Herbal ; pharmacology ; Fatty Acids, Nonesterified ; metabolism ; Fatty Liver ; metabolism ; Lipid Metabolism ; Lipid Peroxidation ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Transcriptome ; drug effects ; Triglycerides ; metabolism
10.Effect of CKJ recipe containing serum on activation of rat primary hepatic stellate cells, TGF-beta1 and its receptors.
Liang CHEN ; Qin FENG ; Jing-hua PENG ; Lin LIU ; Chun-geng LIANG ; Ya-mei HAI ; Yi-yang HU
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(2):210-215
OBJECTIVETo observe the effect of CKJ Recipe (consisting of Cordyceps sinensis polysaccharide, amygdaloside, and gypenosides) containing serum on the activation of rat primary hepatic stellate cells (rHSCs) and to explore its pharmacological mechanism.
METHODSrHSCs were isolated form liver and cultured for four days. Then they were divided into the normal control group, the model group, and the CKJ group. rHSCs in the model group and the CKJ group were treated with 2.5 ng/mL transforming growth factor beta1 (TGF-beta1) in serum-free DMEM for 24 h. Serum free DMEM (containing no TGF-beta1) was taken as the control for the normal control group. rHSCs in the CKJ group were treated with 5% CKJ-containing serum for 24 h. rHSCs in the other two groups were treated with 5% blank serum for 24 h.The protein expression level of a smooth muscle actin (alpha-SMA) was determined using high throughput screening (HCS) and Western blot. mRNA expression levels of alpha-SMA, collagen I (Col-I), platelet-derived growth factor receptor beta (PDGF-betaR), TGF-beta1, transforming growth factor beta receptor 1 (TGF-betaR1), and transforming growth factor beta receptor 2 (TGF-beta R2) were detected using quantitative RT-PCR.
RESULTSCompared with the normal control group, the protein expression level of alpha-SMA, mRNA expression levels of alpha-SMA, Col-I, PDGF-betaR, TGF-beta1, TGF-betaR1, and TGF-betaR2 significantly increased in the model group (P<0.05, P<0.01). Compared with the model group, the protein expression level of alpha-SMA, mRNA expression levels of alpha-SMA, Col-I, PDGF-betaR, TGF-beta1, TGF-beta1, and TGF-beta R2 significantly decreased in the CKJ group (P<0.05, P<0.01).
CONCLUSIONCKJ containing serum could inhibit the protein expression level of o-SMA, which was probably related with inhibiting TGF-beta1 and its related receptors.
Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hepatic Stellate Cells ; metabolism ; Rats ; Transforming Growth Factor beta ; Transforming Growth Factor beta1 ; metabolism