3.Inflammatory reaction changes with aging in kidneys of human TIMP-1 transgenic mice
Xue-Guang ZHANG ; Xiang-Mei CHEN ; Quan HONG ; Xi-Yao SHANG ; Suo-Zhu SHI ; Zhong YIN ; Guang-Yan CAI
Chinese Journal of Geriatrics 2003;0(12):-
Objective To explore the role of tissue inhibitor of metalloproteinase-1(TIMP-1) during renal senescence by using human TIMP-1 transgenic mice.Methods Renal histological changes of wild type mice and transgenic mice at the age of 3,12,24 months were observed by periodic acid-schiff(PAS)staining of paraffin sections.The numbers of F4/80 positive cells were detected by immunofluoreseence.The protein expressions of TIMP-1,TIMP-2,matrix metalloproteinase(MMP)-9,MMP-2,intercellular adhesion molecule-1(ICAM-1),transforming growth factor?1(TGF-?1),collagenⅢand collagenⅣwere detected by Western blot.The activities of gelatinases and TIMP-1 were examined by gelatin zymography and reverse zymography respectively.Results Focal renal fibrosis was found in two genotypes with aging.At the age of 24 months,compared with wild type,in kidneys of transgenic type,the expressions and activities of gelatinases were dowregulated (MMP-2:2.08?0.20 vs.3.39?0.43;MMP-9:4.02?0.82 vs.6.72?1.40,all P<0.05);the expressions of collagenⅢ,collagenⅣ,ICAM-1,and TGF-?1 were upragulated(0.72+0.11 vs.0.57?0.09;0.84?0.13 vs.0.6?0.11,0.72?0.12 vs.0.53?0.07; 0.69?0.12 vs.0.45?0.09,all P<0.05),and the numbers of F4/80 positive cells were increased (18.8?4.4 vs.12.7?3.6,P<0.05)with the upregulated expression and activity of TIMP-1(1.10?0.18 vs.0.62?0.09;50.75?7.25 vs.20.64?3.50,P<0.05).Conclusions TIMP-1 could promote age-related renal fibrosis through enhancing inflammation reaction by ICAM-1 upregulation.
4.Soluble high-expression, purification and bioassay of IGFBP-3.
Chen WU ; Guang-Yin YAO ; Min-Ji ZOU ; Guang-Yu CHEN ; Min WANG ; Jia-Xi WANG ; Dong-Gang XU
Chinese Journal of Biotechnology 2007;23(3):398-402
cDNA for Insulin-like growth factor binding protein 3 was cloned and constructed a prokaryotic expression vector--pET-DsBA-IGFBP3. The construct was transformed into E. coli BL21 (DE3)plysS. The induced fusion protein (D-IGFBP3) was expressed successfully in soluble form. We obtained D-IGFBP3 the purify of which is over 95% after purification by His affinity chromatography. The product was identified by Western-blot. The cell assay showed that the obtained fusion protein can inhibit the growth of MCF-7 and bind with IGF-I in vitro.
Blotting, Western
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Cell Line, Tumor
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Cell Proliferation
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drug effects
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Chromatography, Affinity
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Dose-Response Relationship, Drug
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Electrophoresis, Polyacrylamide Gel
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli
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genetics
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Gene Expression
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Humans
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Insulin-Like Growth Factor Binding Protein 3
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genetics
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metabolism
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pharmacology
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Insulin-Like Growth Factor I
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metabolism
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Protein Binding
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Recombinant Proteins
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isolation & purification
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metabolism
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pharmacology
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Solubility
5.Effect of pinggan qianyang on hypothalamic proteome in the hyperthyroid rats with hyperactivity of liver-yang.
Yao-hui YIN ; Zhen-jia YI ; Guang-wei ZHONG ; Jian-jun HU
Journal of Central South University(Medical Sciences) 2008;33(5):391-398
OBJECTIVE:
To investigate the effect of pinggan-qianyang (PGQY), a Chinese medicine, on hypothalamic proteome in the hyperthyroid rats with hyperactivity of liver-yang, and to explore its mechanism.
METHODS:
The rat model was established by intraperitoneal injection of levo-thyroxine (L-T4) and fuzi decotion. All the quantitative and qualitative changes of the protein expressions were compared among the normal group,the model group and the treatment group by proteomic techniques.
RESULTS:
The protein spots in the 3 groups were mainly displayed at the isoelectric point (pI) 3 approximately 10, and the molecular weights were 13.8 approximately 98.8 kD.Compared with the normal group, 6 spots of protein expression increased and 10 decreased in the model group. All the changed protein in the model group returned to normal level after PGQY treatment. Mass-spectrometer and bio-informatics indicated that these proteins were Prohibitin, Peroxiredoxin-6, histidine triad nucleotide-binding protein 1, protein-tyrosine-phosphatase, predicted protein, profilin-2, peroxir doxin-II, heat shock protein-27, and annexin-A1.
CONCLUSION
There are differences in the expression of hypothalamus proteins in the hyperthyroid rats with hyperactivity of liver-yang after the treatment with PGQY, and the 9 identified protein spots may be associated with the mechanism of PGQY.
Animals
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Drugs, Chinese Herbal
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therapeutic use
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Female
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Hyperthyroidism
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drug therapy
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metabolism
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Hypothalamus
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metabolism
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Male
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Medicine, Chinese Traditional
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Nerve Tissue Proteins
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metabolism
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Peroxiredoxin VI
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metabolism
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Phytotherapy
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Proteome
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metabolism
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Random Allocation
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Rats
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Rats, Sprague-Dawley
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Repressor Proteins
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metabolism
6.Evaluation of the effects of glycosylation on in vivo survival of cold-storage human platelets by using rabbit model.
Rong-Cai TANG ; Cheng-Yin HUANG ; Xiao-Ping PEI ; Bao-An CHEN ; Cui-Ping LI ; Guang-Yao SHI
Journal of Experimental Hematology 2005;13(6):1113-1116
To study the effects of glycosylation on survival of cold-storage human platelets by using rabbit model. (51)Cr-labeling platelets were used to detect the platelet storage survival. The human platelets (2.0 x 10(12)/L) treated with 5 g/L uridine diphosphate galactose (UDP-Gal) were stored in 4 degrees C refrigeratory up to 10 days. The survival of human platelets in rabbits whose reticuloendothelial system was inhibited by the administration of ethyl palmitate was monitored in blood drawn at various times after the platelet transfusion. The results showed that the survival rate of platelets was significantly increased in cold-storage human platelets by UDP-Gal treatment. The survival rates of platelets at 2 hours after transfusion into rabbits in groups of fresh platelets group, UDP-Gal + cold platelets group and cold platelets group were (68.9 +/- 8.5)%, (65.4 +/- 8.0)% and (5.0 +/- 2.6)%, respectively. Compared with cold platelets group, significant differences were seen among all groups (P < 0.01). UDP-Gal + cold platelets group had no significant differences compared with fresh platelets group (P > 0.05). It is concluded that UDG-Gal can provide the protective effect on cold-storage human platelets and prolong the survival time of refrigerated human platelets in rabbit model.
Animals
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Blood Platelets
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cytology
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metabolism
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Blood Preservation
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Cell Survival
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drug effects
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Cryopreservation
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methods
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Glycosylation
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drug effects
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Humans
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Models, Animal
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Platelet Transfusion
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Rabbits
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Uridine Diphosphate Galactose
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pharmacology
7.Application of cationic propyl gallate as inducer of thrombocyte aggregation for evaluating the platelet function of platelet donors.
Da-Xiang SHENG ; Cheng-Yin HUANG ; Guang-Yao SHI ; Xi-Lin OUYANG ; Li CAI ; Jian-Yu XIAO ; Rong-Cai TANG
Journal of Experimental Hematology 2005;13(6):1099-1102
The purpose of study was to investigate the feasibility of the application of cationic propyl gallate (C-PG) as inducer of platelet aggregation for evaluating the platelet function of single-donor plateletpheresis and identifying the incidence of defective platelet function among donors. Experiments were as follows: 3 healthy volunteers' platelet aggregation induced by 100-300 micromol/L C-PG was determined by LG-PABER analyzer to observe the effect of C-PG concentration on platelet aggregation; 30 healthy volunteers' platelet aggregation before and 24 hours after administration of 200-400 mg acetylsalicylic acid (ASA) was examined after induction by 200 micromol/L C-PG for determining the cut-off value to discriminate platelet dysfunction donors; the platelet aggregation of 483 platelet donors was detected and the activated plasma clotting time (APCT) of donors who have deficiency in platelet aggregation was examined for investigating the incidence of defective platelet function among donors. The results showed that platelets were activated by C-PG induction in a dose dependent manner, when concentration of C-PG reached 200 micromol/L, the percentage of platelet aggregation was highest. It significantly decreased after 24 hours with ASA than that before the administration (P < 0.001), especially in 180 seconds induced by C-PG. If cut-off point was fixed on the platelet aggregation < 20% in 180 seconds, donors of platelet dysfunction can be selected effectively. 25 of defective platelet aggregation function among 483 donors were detected, and 11 out of 25 platelet dysfunction donors had the deficiency in procoagulant activity with prolonged APCT. It is concluded that C-PG as inducer of platelet aggregation is feasible to screen the platelet function of donors. Five percent of platelet donors has function defect examined by C-PG as inducer of platelet aggregation.
Antioxidants
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chemistry
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pharmacology
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Aspirin
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administration & dosage
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Blood Donors
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Blood Platelets
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cytology
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drug effects
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physiology
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Cations
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chemistry
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Humans
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Platelet Activation
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drug effects
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Platelet Aggregation
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drug effects
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Platelet Aggregation Inhibitors
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administration & dosage
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Platelet Function Tests
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Platelet Transfusion
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Propyl Gallate
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administration & dosage
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chemistry
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Whole Blood Coagulation Time
9.Effect of Weikangfu granule on the physiopathologic figure of precancerosis of gastric mucosa in patients of chronic gastritis with Pi-deficiency syndrome.
Guang-yao YIN ; Yi CHEN ; Wu-ning ZHANG ; Xiao-jing SHEN ; Jian-hua JIAO ; Xue-fen HE
Chinese journal of integrative medicine 2005;11(4):264-271
OBJECTIVETo study the physiopathologic basis of Weikangfu Granule (WKFG) in treating precancerosis of gastric mucosa in patients of chronic gastritis with Pi-deficiency syndrome (CG-PDS).
METHODSOne hundred and fifteen patients of CG-PDS who suffered from intestinal metaplasia (IM) and atypical hyperplasia (ATHP) of gastric mucosa, were divided into two groups. The treated group (n = 61) was treated by WKFG with its ingredients modified according to the syndrome type of patients. The control group (n = 54) was treated with Weishu granule. The histopathological and subcellular ultrastructural changes were detected by optical microscope, screening electronic microscope, transmission electronic microscope and histochemical staining; the nuclear and mitochondrial ultrastructure of gastric mucosa were analyzed with energy dispersion X-ray analyser and image analysis system. And the changes of cAMP, lipid peroxide (LPO), superoxide dismutase (SOD) before and after treatment in the treated group were measured and compared with those of the health control group consisting of 15 volunteers.
RESULTSThe symptomatic and pathological therapeutic effect in the treated group were significantly superior to those in the control group (P < 0.05). The contents of Zn, Cu, cAMP, SOD and (3)H-TdR LCT in gastric mucosa of the treated group before treatment were all lower than those of the healthy control group, yet all these indexes markedly increased after treatment, while serum LPO level, which increased before treatment was lowered after treatment. All the changes showed statistical significance (P < 0.05 or P < 0.01).
CONCLUSIONWKFG can reverse IM and ATHP in patients of CG-PDS, and the effect may be realized by way of increasing the level of Zn, Cu, cAMP and SOD in gastric mucosa, promoting cell differentiation, enhancing cellular immunity and reducing oxygen free radicals and lipid peroxidation.
Adult ; Aged ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Chronic Disease ; Copper ; analysis ; Cyclic AMP ; analysis ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; chemistry ; pathology ; ultrastructure ; Gastritis, Atrophic ; pathology ; Humans ; Lipid Peroxides ; analysis ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Precancerous Conditions ; pathology ; Stomach Neoplasms ; pathology ; Superoxide Dismutase ; analysis ; Syndrome ; Yang Deficiency ; complications ; Zinc ; analysis
10.Study on the pathophysiologic basis of classification of 'spleen' deficiency in chronic gastritis.
Guang-yao YIN ; Yi CHEN ; Xiao-jing SHEN ; Xue-fen HE ; Wu-ning ZHANG
Chinese Medical Journal 2005;118(6):468-473
BACKGROUNDMost of the studies on traditional Chinese medicine (TCM) 'spleen' deficiency syndrome in the recent 30 years were conducted only on the basis of single functional index, neglecting the study on the pathophysiologic internal relationship between spleen deficiency syndrome and gastric diseases in modern medicine. But it was at the subcellular molecular biological level that we explored the pathophysiologic basis of classification of spleen deficiency in chronic gastritis by detecting the bioactive substances in gastric mucosa nuclei and mitochondria.
METHODSBy means of optical microscope, scanning electron microscope (SEM), transmission electron microscopy (TEM) and histochemical staining, we conducted histopathological, subcellular ultrastructural analysis and nuclei and mitochondrial ultrastructural analysis of gastric mucosa of 188 spleen deficiency patients and of 42 voluntary blood donors. At the same time, bioactive substances were measured by means of X-ray energy dispersive analysis system (EDAX) image analysis system, radioimmunoassay method and chemiluminescence method.
RESULTSThe content of cAMP, superoxide dismutase (SOD), Zn and Cu in gastric mucosa, and the content of Zn and Cu in mitochondria decreased progressively in order of groups: healthy control (HC), spleen Qi deficiency without organic lesion (F-SQD), spleen Yang deficiency without organic lesion (F-SyangD), disease without symptoms group, spleen Qi deficiency with organic lesion (G-SQD), spleen Yang deficiency with organic lesion (G-SyangD), spleen Yin deficiency (SyinD) and spleen deficiency with Qi stagnation (SDQS), chronic spleen deficiency gastritis (CSG) and chronic atrophic gastritis (CAG); decreased in order of HC, intestinal metaplasia (IM)Ia, IMIb, IMIIa and IMIIb, P < 0.05. The content of DNA, Zn and Cu in nuclei progressively increased in order mentioned above, P < 0.05.
CONCLUSIONSThe quantitative changes of gastric mucosal cAMP, SOD, Zn, Cu, of mitochondrial Zn, Cu and of nuclear DNA, Zn and Cu are not only the substance base on which the lesion of gastric mucosa tissue structure occurs, but also the substance base on which spleen deficiency is classified. G-SQD and G-SyangD were more likely to be found in low-grade or middle-grade CSG and CAG, while SyinD and SDQS in middle-grade or high-grade CSG, CAG and IMIIb.
Adult ; Aged ; Chronic Disease ; Cyclic AMP ; analysis ; Female ; Gastric Mucosa ; pathology ; ultrastructure ; Gastritis ; metabolism ; pathology ; Humans ; Lipid Peroxides ; blood ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Splenic Diseases ; classification ; Superoxide Dismutase ; analysis