2.Effects and possible role of recombinant human erythropoietin on endothelial progenitor cells from patients with diabetic nephropathy
Shi ZHAO ; Hongxiang WANG ; Shiying SHAO ; Bingong LI ; Hong MAO
Chinese Journal of Postgraduates of Medicine 2008;31(28):12-15
Objective To study the effect and possible role of recombinant human erythropoietin(rhEPO) on the proliferation and apoptosis of endothelial progenitor cells (EPCs) from patients with diabetic nephropathy (DN). Methods Various concentration of rhEPO was added to the culture system of EPCs from 20 DN patients (DN group) and 20 normal people (control group). MTT assays were used to detect the proliferative rate, Annexin V/PI stains to detect the apoptotic rate, and Western blot assays to detect the expression level of Akt protein kinase. Results Proliferative ability of EPCs from control group and DN group was improved when concentration of rhEPO was 0.3, 0.6 and 1.2 kU/L, and it was dose-dependent. The effect from the latter was more obvious. The apoptotic rate of DN group was lowered and the expression levels of Akt protein kinase were upregulated when the concentration of rhEPO was 1.2 kU/L, while this kind of effect was blocked after Wortmannin was added to the culture system. Conclusion rhEPO can improve the number and function of EPCs from both healthy volunteers and patients with DN. PI3K/Akt pathway may play an important role.
3.Effects of high-glucose on proliferation and apoptosis in endothelial progenitor cells of type 2 diabetes mellitus
Hongxiang WANG ; Shi ZHAO ; Bingong LI ; Hong MAO ; Shiying SHAO
Chinese Journal of Pathophysiology 2000;0(11):-
AIM: This study aimed to observe the effects of high-glucose on proliferation and apoptosis of endothelial progenitor cells (EPCs) in type 2 diabetes mellitus patients, and tried to elucidate their possible role. METHODS: Various concentrations of glucose were added to the culture system of EPCs from 25 cases of type 2 diabetes mellitus patients (DM group) and 25 cases of healthy volunteers (control group). MTT assays were used to detect the proliferative rates. Annexin-V/PI stains were used to detect the apoptotic rates, and RT-PCR to detect the expression level of bcl-2 and bax. RESULTS: Proliferative activity of EPCs in both control group and DM group were attenuated when concentration of glucose was 33 mmol/L, while apoptotic rates increased. No significant change of proliferative rate and apoptotic rate of EPCs in DM group and control group in the presence of 5 mmol/L glucose was observed. The expression level of bax of EPCs in both DM group and control group increased while expression level of bcl-2 did not change much in the presence of 33 mmol/L glucose. CONCLUSION: High-glucose attenuates proliferative activity of EPCs and increases the apoptotic rate. Upregulation of bax may be its possible role.
4.Comparison of Clinical Efficacy of Lyophilized Recombinant Human Brain Natriuretic Peptide and Dobutamine in Treatment of Acute Heart Failure and Plasma Gal-3, CysC and ET-1 Levels
Zhen ZHU ; Jianrong ZHAO ; Ziyang LI ; Qiang LU ; Hong SHAO
Progress in Modern Biomedicine 2017;17(26):5145-5148,5156
Objective:To explore and compare the clinical efficacy of lyophilized recombinant human brain natriuretic peptide (Lrh-BNP) and dobutamine (Dob) in the treatment of patients with acute heart failure (AHF) and impacts on the plasma galectin (Gal)-3,Cystatin C (CysC) and endothelin (ET-)-1 levels.Methods:114 cases of patients with AHF in our hospital from February 2015 to February 2017 were selected as the research objectives and randomly divided into two groups.Dob group was treated by Dob,while Lrh-BNP group was treated by Lrh-BNP.The cardiac function parameters,plasma Gal-3,CysC,ET-1 levels before and after treatment,clinical comprehensive efficacy and incidence of adverse reactions were compared between two groups.Results:The FS,LVEF levels of both groups at 72 hours after treatment were significantly higher than those before treatment (P<0.01),but the LVEDD,plasma Gal-3,CysC,ET-1 levels were obviously decreased (P<0.01),the index mentioned above of Lrh-BNP group improved more significantly than those of the Dob group(P<0.01).The overall effective rate of Lrh-BNP group was 89.5 %,which was significantly higher than that of the Dob group (73.7%,P<0.05).No significant difference was found in the incidence of adverse reaction between two groups(P>0.05).Conclusion:Lyophilized recombinant human brain natriuretic peptide was more effective in the treatment of AHF than Dobutamine with equal safety,which might be related to the decrease of plasma Gal-3,CysC,ET-1 levels.
5.The Radiosensitizing Effect of Resveratrol on Hopypharyngeal Carcinoma Cell Line FADU and its Effect on the Cell Cycle.
Yuan SHAO ; Fang QUAN ; Hong-hui LI ; Xiao-bao YAO ; Qian ZHAO ; Rui-min ZHAO
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(6):699-703
OBJECTIVETo study the radiosensitizing effect of resveratrol on hypopharyngeal carcinoma cell line FADU in vitro.
METHODSHypopharyngeal carcinoma cell line FADU was cultured in in vitro DMEM. Its inhibition on cell proliferation was detected using cytotoxicity test (MTT assay). The cell survival curve was drawn using clone formation to obtain sensitive enhancement ratio (SER). Changes of the cell cycle and cell apoptosis were analyzed using flow cytometry (FCM).
RESULTSResults of MTT showed the inhibition of resveratrol on FADU cells increased along with its concentrations (P < 0.05). Results of clone formation indicated the surviving fraction at 2 Gy (SF2) was 0.717 ± 0.062 in the irradiation group, and 0.426 ± 0.035 in the resveratrol plus irradiation group (with SER ranged 1.684 ± 0.178) with statistical difference (P = 0.007). Results of FCM showed that after radiation of 4 Gy radiation, cells at G2/M phase arrest increased, but cells at G1 decreased. After radiation of resveratrol for 24 h, cells at G1 decreased, but cells at G2/M phase and S phase arrest increased. When 4 Gy radiation combined resveratrol was used, cells at G2/M phase arrest significantly increased, but cells at G1 significantly decreased. The apoptosis rate was 1.94% ± 1.65% in the control group, 4.56% ± 0.92% in the irradiation group, 2.03% ± 1.46% in the resveratrol group, and 23.11% ± 7.22% in the resveratrol plus irradiation group. There was statistical difference between the resveratrol plus irradiation group and the rest 3 groups (P < 0.05).
CONCLUSIONResveratrol could enhance the radiosensitivity of hypopharyngeal carcinoma FADU cells in vitro possibly by inducing cell apoptosis and causing changes in the cell cycle distribution.
Apoptosis ; Carcinoma, Squamous Cell ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; Head and Neck Neoplasms ; Humans ; Hypopharyngeal Neoplasms ; drug therapy ; Radiation Tolerance ; Radiation-Sensitizing Agents ; therapeutic use ; Stilbenes ; therapeutic use
6.Biological characteristics of Fusarium oxysporum and inhibitory effects of five fungicides.
Qing-Song SHAO ; Hong-Bo LIU ; Xiao-Fang ZHAO ; Run-Huai HU ; Ming-Yan LI
China Journal of Chinese Materia Medica 2014;39(8):1386-1390
The mycelium growth and sporulation in different temperature, pH value and light conditions were detected by using the crossing and haemocytometer method. The toxicity of five fungicides to Fusarium oxysporum was tested by mycelia growth method, so as to provide the reference for prevention and control against F. oxysporum. The optimum temperature and pH value of F. oxysporum to grow and spore were 28 degrees C and 6-7. Alternating light and darkness promoted growth and sporulation of bacterial colony. As for five fungicides, the EC50 of tebuconazole was 10.02 mg x L(-1), 92.50 times as much as carbendazim. The EC50 of myclobutanil and Fuxing was 91.23, 96.68 mg x L(-1), respectively. Tebuconazole showed the most tremendous inhibitory effect and control efficiency on F. oxysporum.
Fungicides, Industrial
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pharmacology
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Fusarium
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drug effects
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growth & development
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Mycelium
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drug effects
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growth & development
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Orchidaceae
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microbiology
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Plant Diseases
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microbiology
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Spores, Fungal
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drug effects
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growth & development
7.Establishment and application of a high-throughput drug screening model based on COL1A1 promoter for anti-liver fibrosis.
Shuang-Shuang ZHAO ; Ju-Xian WANG ; Yu-Cheng WANG ; Rong-Guang SHAO ; Hong-Wei HE
Acta Pharmaceutica Sinica 2015;50(2):169-173
For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I
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genetics
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Drug Evaluation, Preclinical
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methods
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Genes, Reporter
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Hepatic Stellate Cells
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High-Throughput Screening Assays
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Humans
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Liver Cirrhosis
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drug therapy
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Luciferases
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Plasmids
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Promoter Regions, Genetic
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RNA, Messenger
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Transfection
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Transforming Growth Factor beta1
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pharmacology
8.Determination of five pyrethroid pesticides residues in Anoectochilus roxburghii by cloud point extraction-back extraction and GC-MS.
Hong-Bo LIU ; Xiao-Fang ZHAO ; Gui-Ying SHI ; Qing-Song SHAO ; Mine-Van LI
China Journal of Chinese Materia Medica 2014;39(15):2859-2862
A method for residual determination of 5 pyrethroid pesticides in Anoectochilus roxburghii by cloud point extraction-back extraction-GC-MS was established. PEG 6000 was used as extraction agent and isooctane was used for back-extractant. The con- tent was calculated by external standard method. The linear range was from 15 to 2 000 μg x kg(-1) with the good correlation coefficients (0.955-0.999). The recoveries at spiked concentrations of 50-500 μg x kg(-1) ranged from 85.12% to 101.6%. The limit of detection and quantification of 5 pyrethroid pesticides were in the range of 0.63-3.10 μg x kg(-1) and 2.10-10.31 μg x kg(-1), respectively. The proposed method can be applied to the determination of pyrethroid pesticides residues in A. roxburghii.
Chemical Fractionation
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methods
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Gas Chromatography-Mass Spectrometry
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methods
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Orchidaceae
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chemistry
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Pesticide Residues
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analysis
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chemistry
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isolation & purification
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Pyrethrins
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analysis
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chemistry
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isolation & purification
9.Quantitative analysis of renal injury using CEUS in hypertensive rabbits model induced by L-NAME
Xianglu MENG ; Guohui ZHANG ; Wenhong WANG ; Hong SHAO ; Jun GUO ; Xiaoxin ZHAO ; Cheng GU ; Xiaodan ZHANG
Tianjin Medical Journal 2015;(12):1394-1397
Objective To assess the value of contrast-enhanced ultrasound (CEUS) on quantitative analysis of re?nal cortex perfusion in hypertensive rabbits model. Methods Hypertensive rabbit modal (n=10) were established by inject?ing N-nitro-L-arginin methylester (L-NAME). CEUS and Cystatin C (CysC) serum level analysis were performed at differ?ent time points:before and the 2nd, 4th, 6th and 8th week after injecting L-NAME. Time-intensity curve and area under curve (AUC) were analyzed quantatively while correlation of AUC and CysC were also analyzed. Results Serum level of Cys C in?creased significantly at the 6th week after L-NAME administration which is earlier than the increase of serum levels of Scr and BUN. AUC decreased at first then increased after L-NAME administration. Upon addition of L-NAME, rise time (RT) and peak intensity (PI) decreased while mean transit time (MTT), time from peak to one half (HPT) and time to peak (TTP) in?creased. Our study confirmed a positive correlation between AUC and Cys C (r=0.950, P<0.001). Conclusion Setting up rabbits model by L-NAME is convenient and reproducible, which is an useful tool in experimental study of preclinical and clinical phase of hypertensive renal injury. CEUS combining with CysC serum level analysis is considered as an effective technology for evaluating renal function in hypertensive patients.
10.Effect of bone mesenchymal stem cells on lidocaine-induced apoptosis in rat dorsal root ganglion cells in vitro
Xuejun SUN ; Yan SHAO ; Hongxing ZHANG ; Fang ZHOU ; Xi ZHAO ; Hong MA
Chinese Journal of Anesthesiology 2013;33(9):1076-1078
Objective To evaluate the effect of bone mesenchymal stem cells (BMSCs) on lidocaine-induced apoptosis in dorsal root ganglion cells (DRGCs) of rats in vitro.Methods DRGCs in the logarithmic phase were incubated in culture plates at the density of 2 × 104 cells/cm2 (27 wells in total).DRGCs were randomly divided into 3 groups (n =9 each) using a random number table:control group (group C),lidocaine treatment group (group L) and BMSC treatment group (group B).The DRGCs in group C were incubated routinely without lidocaine,while the DRGCs were incubated for 2 h with lidocaine with the final concentration of 50 mmol/L in L and B groups.The DRGCs were then incubated normally in group L.The DRGCs were then co-cultured with the BMSCs which were incubated in Transwell chambers with the density of 2 × 104 cells/cm2 in group B.DRGCs were collected at 48 h of incubation for detection of apoptosis by flow cytometry.Apoptosis rate was calculated.Results Compared with group C,the apoptosis rate was significantly increased in L and B groups (P < 0.05).The apoptosis rate was significantly lower in group B than in group L (P < 0.05).Conclusion BMSCs can reduce lidocaine-induced apoptosis in DRGCs of rats in vitro,indicating that BMSCs may reduce local anesthetics-produced toxicity to the peripheral nerve.