1.Expression of Matrix Metalloproteinase-9/Tissue Inhibitor of Metalloproteinase-1 in Rats with Tubulointerstitial Fibrosis
yan, WANG ; de-an, ZHAO ; da-sheng, YANG
Journal of Applied Clinical Pediatrics 1992;0(05):-
Objective To observe the expression of matrix metalloproteinase-9(MMP-9)/tissue inhibitor of metalloproteinase-1(TIMP-1) in tubulointerstitial fibrosis(TIF) rats and explore the effects of MMP-9 and TIMP-1 in TIF.Methods Forty-eight male Sprague-Dawley(SD) rats were randomly divided into sham-operated group(n=24) and model group(n=24).Results Tubulointerstitial fibrosis model was established via unilateral ureteral obstruction(UUO). Histological changes were observed in tubulointerstitium injury under microscope and immunohistochemistry staining was performed to investigate both the expression of MMP-9 and TIMP-1 in TIF by semiquantitative score at d7, d14, d21 after experiment onset.The expressions of TIMP-1 in rats from sham-operated group were lower than those of mo-del group rats at d7, d14, d21 (Pa
6.Influence of high-fluoride on thyroid function and brain damage in rats
Yan-hong, QIU ; De-ming, KONG ; Qin, YANG ; Na, ZHAO
Chinese Journal of Endemiology 2010;29(2):146-149
Objective To study the influence of high-fluoride on thyroid function and brain damage. Methods Thirty-six Wistar rats were randondy divided, according to weight and gender into 3 groups(12 rats each), i.e. control group, high fluoride group, and high fluoride plus thyroid tablet treatment group. The rats were fed with normal tap water containing no more than 5 mg/L NaF and the tap water added 100,100 mg/L NaF, respectively. After 7 months of experiment, the rats in high fluoride plus thyroid tablet treatment group were given with 0.04% thyroid tablet( 1.8 ml·kg~(-1)·d~(-1)) by gastric perfusion for three weeks. The contents of TT_3 and TT_4 in serum were detected by radio-immunological assay; the histomorphology in thyroids and brains were observed under microscopy; and the protein level of NMDAR2B subunit of glutamate receptor in the hippocampal CA1 and CA3 was measured by immunohistochemistry. Results As compared to the values of TT_3 and TT_4 in serum of rats in control group[ (0.97 ± 0.15), (84.03 ± 12.45)nmol/L], TT_3 and TT_4 in high fluoride group were obviously lower [(0.24 ± 0.07), (15.16 ± 2.08)nmol/L, all P < 0.01]; while no changes in TT_3 and TT_4 were detected in high fluoride plus thyroid tablet treatment group[ (1.02 ± 0.19), (85.63 ± 9.55)nmol/L, all P < 0.05] as compared to controls, but higher than those in high fluoride group(all P < 0.01 ). The pathological changes including partial hyperplasy, arrangement disorder, atrophy, and decreased colloid of the thyroid follicular epithelial cells in high fluoride group were observed under microscopy. In high fluoride plus thyroid tablet treatment group, the degree of the thyroid cellular hyperplasy was relatively slight as compared to high fluoride group. The swelling and disarrangement of neurons in the hippocampus were observed in high fluoride group, whereas the changes of the neurons were not so obvious in high fluoride plus thyroid tablet treatment group. The grey values of NMDAR2B positive cells in the hippocampal CA1 and CA3 in high fluoride group(167.05 ± 7.31 ) were significantly increased as compared to controls (92.53 ± 9.67 ) or high fluoride plus thyroid tablet treatment group( 101.66 ± 12.21, all P < 0.01 ). Conclusions High fluoride can induce the decreased function and changed histomorphology in thyroid and result in pathological damages in the brains of rats. However, treated with thyroid tablet to those having damages induced by high fluoride, the thyroid function and morphology can be normal, and the brain damages can be alleviated. The results indicate that hypothyroidism caused by high fluoride might be an important participating factor in brain damages caused by fluorosis.
7.Preliminary Study on Serum Metabolites of Primary Biliary Cholangitis by Hydrogen Spectrum Nuclear Magnetic Resonance
Jing YANG ; De DUAN ; Huamei WU ; Lihong YANG ; Yan LI ; Jinhui YANG
Chinese Journal of Gastroenterology 2017;22(2):75-81
Hydrogen spectrum nuclear magnetic resonance (1 H-NMR ) is a commonly used method for metabolomics and has been applied in the study of liver cirrhosis and liver cancer,however,study on serum metabolites in patients with primary biliary cholangitis (PBC)is rare.Aims:To investigate the capability of 1 H-NMR for screening serum metabolites of PBC.Methods:Twenty PBC patients,20 HBV-related cirrhosis patients and 20 healthy controls were detected by 1 H-NMR.The 1 H-NMR spectra data were processed by principal component analysis (PCA)and orthogonal partial least squares-discriminant analysis (OPLS-DA)so as to create the diagnostic model.Based on the correlation coefficient P (corr),VIP value and non-paired t test of OPLS-DA model,the differential metabolites between normal group and the two cirrhosis groups were screened.Results:OPLS-DA model could effectively distinguish healthy controls and PBC patients,model interpretation ability and prediction ability were 81.9% and 44.8%,respectively (P=0.0293), glutamine,folic acid,urocanic acid,4-ethylbenzoic acid were differential metabolites.OPLS-DA model could also effectively distinguish healthy controls and HBV-related cirrhosis patients, urocanic acid, 1-methylhistamine, 1-methyladenine,glucose,L-acetylcarnitine were differential metabolites.OPLS-DA model could not effectively distinguish PBC patients and HBV-related cirrhosis patients.Conclusions:Serum glutamine and folic acid may be the potential biomarkers of PBC,which may be closely related to the immune damage mechanism and prognosis of PBC;1 H-NMR combined with OPLS-DA diagnostic model are expected to become a new method for studying liver cirrhosis.
8.Determination of unsaturated disaccharides produced from chondroitin sulfate in urine using high performance liquid chromatography with fluorometric post column derivatization
Wei, DONG ; De-an, LI ; Xiao-yan, FU ; Yan-hong, CAO ; Yang, HUI
Chinese Journal of Endemiology 2008;27(3):323-325
Objective To explore the effectiveness of the unsaturated disaccharides derived from chondroitin sulfate(CS)in urine using post column derivation fluorometric method of high performance liquid chromatography (HPLC).Methods Urine samples were prepared with the centrifugal flirtation and the routine precipitation methods separately,then digest the sample with CS lyses,use the post column derivation method of HPLC to fluorometric determine the unsaturated disaecharides(△Di-0S,△Di-4S and △Di-6S),compare the recovery differences of the CS standard in the urine pretreated by the two methods.Results The HPLC system ran well,the lowest detection limit of the method was 10 ng.When the urine was pretreated by the centrifugal flirtation method,the recovery of △Di-0S,△Di-4S and △Di-6S produced from the CS standard were 86.93%,87.28%and 85.03%,respectively,while the recovery of the three disaccharides in routine precipitation method were 75.09%,72.96%and 77.70%merely.Conclusions The HPLC post column derivatization fluorometric method is comparatively sensitive,it can be used to determine unsaturated disaccharides decomposed by CS.In the pretreatment procedure of the urine,the centrifugal flirtation method was superior to the routine precipitation method,it can be used as microdetermination for the unsaturated disaccharides.
9.Stretch-induced Expression of CYR61 Increases the Secretion of IL-8 in A549 Cells via the NF-κβ/Iκβ Pathway
Yan ZHANG ; Ping GUI ; Shang-Long YAO ; Dong YANG ; Yang LV ; De-Fang DING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2018;38(4):672-678
Mechanical ventilation (MV) with large tidal volumes can increase lung alveolar permeability and initiate inflammatory responses,resulting in ventilator-induced lung injury (VILI).The mechanisms of the injurious effects of MV and the genetic susceptibility remain unclear.VILI-related genes such as cysteine-rich angiogenic inducer 61 (Cyr61)have been demonstrated to play a detrimental role in the aggressive ventilation strategies.In the present study,we investigated the involvement of Cyr61 in the VIM and the underlying mechanism.A549 cells were exposed to cyclic stretch of varying durations and then the mRNA and protein levels of Cyr61 were measured by real-time PCR and Western blotting,respectively.Additionally,after exposure ofA549 cells to cyclic stretch for 5 min to 1 h,the expression levels of nuclear factor kappaB (NF-κB) and IL-8 were detected by ELISA and Western blotting.Thereafter,Cyr61 expression was depressed in A549 cells with the siRNA pGenesill.1-Cyr61-3 before the cyclic stretch,and IL-8 secretion and the activation of NF-κB pathways were probed by ELISA and Western blotting,respectively.Moreover,a NF-κB inhibitor (PDTC) and an activator (TNF) were used before mechanical stretch.Realtime PCR and ELISA were performed to detect the mRNA and protein of IL-8,respectively.The results showed that the mechanical cyclic stretch led to increased Cyr61 expression at mRNA and protein levels in A549 cells.Additionally,cyclic stretch also mobilized NF-κB from the cytoplasm to the nucleus and increased IL-8 secretion in A549 cells.The inhibition of Cyr61 blocked the NF-κB activation and IL-8 secretion in response to cyclic stretch.Inhibition of NF-κB attenuated the mRNA and protein expression of IL-8 in A549 cells transfected with Cyr61 siRNA.It was suggested that Cyr61/NF-κB signaling pathway mediates the upregulation of IL-8 in response to cyclic stretch in A594 cells.These findings support the hypothesis that Cyr61 plays a critical role in acute lung inflammation triggered by mechanical strain.
10.Rapid identification of chemical composition in safflower with UHPLC-LTQ-Orbitrap.
Song-song WANG ; Yan MA ; Yi ZHANG ; De-feng LI ; Hong-jun YANG ; Ri-xin LIANG
China Journal of Chinese Materia Medica 2015;40(7):1347-1354
The UHPLC-LTQ-Orbitrap high resolution mass spectrometer was used to explore the chemical compositions in safflower. The rapid separation of the compositions was conducted by the UHPLC, following by high resolution full scan and MS2 scan, under the positive and negative ion mode. The chemical formula of compositions were deduced by full scan data in less than 5, then the potential structures were confirmed by the MS2 data. Forty-nine compounds were detected, of which 26 was identified, and 5 compounds was validated by the standard substances.
Carthamus tinctorius
;
chemistry
;
Chromatography, High Pressure Liquid
;
methods
;
Drugs, Chinese Herbal
;
chemistry
;
isolation & purification
;
Molecular Structure
;
Tandem Mass Spectrometry
;
methods