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.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
7.Long-term efficacy comparison between goniosynechialysis and combination of phacoemulsification with goniosynechialysis for the management of chronic angle closure glaucoma
Jing, ZHU ; Wei, ZHAO ; Jun, SHAO ; Xun, BAO ; Jing, LIN ; Dong-hong, FU ; Yong, YAO
Chinese Journal of Experimental Ophthalmology 2013;31(10):964-967
Background One of the features of the pathogenesis of primary angle-closure glaucoma(PACG) is anterior synechia of peripheral iris.Goniosynechialysis and combination of phacoemulsification and goniosynechialysis have been applied for the treatment of the disease recently,but the selection of operative types has great impact on clinical efficacy.Objective This study was to investigate the long-term efficacy of goniosynechialysis and combination of phacoemulsification and goniosynechialysis for the management of chronic PACG.Methods A non-randomized clinical controlled trial was designed.This clinical trial complied with Declaration of Helsinki and was approved by Medical Ethic Committee of Nanjing Medical University.Written informed consent was obtained from each patient.One hundred and ten eyes of 110 patients with chronic PACG were assigned to the goniosynechialysis group and combined operative group from March,2008 to February,2011 in Wuxi People's Hospital.180° goniosynechialysis was performed on 34 patients in the goniosynechialysis group,and phacoemulsifieation +intraocular lens (IOL) implantation + goniosynechialysis were carried out in 78 patients of the combination operative group.All the patients were followed-up for 2 years.Vision acuity,intraocular pressure (IOP),anterior chamber depth(ACD),unltrasound biomicroscopy and perimetry were recorded and compared between before and after operation.Results No significant difference was found in vision acuity between preoperation and postoperative 2 years in the goniosynechialysis group ([0.65 ± 0.15] vs.[0.45 ± 0.15]) (t =1.57,P>0.05),but in the combination operative group,the vision acuity was significantly different between the before and after operation ([0.25±0.15] vs.[0.85 ±0.05]) (t =9.12,P<0.001).The lOPs at 2 years after operation were (14.2±4.1) mmHgand(13.7±4.8) mmHg,respectively in the goniosynechialysis group and combination operative group and were significantly lower than(47.2 ±6.3) mmHg and(46.9±7.0) mmHg before operation(t =4.95,P<0.001 ;t=5.03,P<0.001).The ACD values in the goniosynechialysis group and combination operative group were(3.38±0.02)mm and (3.54±0.03) mm 2 years after operation,which were significantly increased in comparison with (1.33 ±0.24)mm and (1.56±0.37) mm before operation(t=7.65,P<0.001;t=6.76,P<0.001).Conclusions Both combination of phacomulsification with goniosynechialysis or 180° goniosynechialysis are effective for the treatment of PACG.Suitable operation should be alternated depending on the indicators of PACG patients.
8.The association between HLA-A antigen carrying Bw4 epitope and HIV-1 viral loads and CD+4 T cell counts
Jianping CHEN ; Mingming JIA ; Hongwei LIU ; Sha LIU ; Xiaoqing ZHANG ; Hongring ZHAO ; Yiming SHAO ; Kunxue HONG
Chinese Journal of Laboratory Medicine 2009;32(4):383-387
Objective To investigate the impact of HLA-A antigens carrying Bw4 epitopes on disease course of HIV-1 infection.Methods Three hundred and forty subjects chronically infected with HIV-I were recruited and their HLA-A and B alleles were genotyped with sequence-based high resolution typing assay.HLA-Bw genotypes of these HIV-1 infected subjects were determined and their association with CD+4 T cell counts and viral load were analysed.Results When compared with subjects canting no Bw4 epitopes in HLA-A and HLA-B loci (OBw4) (median of CD+4 T cell counts:294/μ1;plasma viral load median 6.29×104 copies/ml),CD+4 T cell counts in subjects with genotypes of 1Bw4-A and 2Bw4-AA were comparable (307 and 308/μ1,respectively),but higher viral load (1.53×105 and 2.68×105 copies/ml,respectively) was observed.In subjects with Bw4 epitopes in HLA-B alleles but no in HLA-A,significantly higher CD+4 T cell counts (417/μ1,P=0.013) and lower viral load (2.10×104 copies/ml,P=0.007) were observed compared with those without Bw4 in HLA-A and HLA-B.Conclusion HLA-B antigens carrying Bw4 epitope were protective in HIV-1 infection by maintaining higher CD+4 T cell counts and lower viral load,but such protective effect was not observed in HLA-A antigens earring Bw4 epitope.
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.Surgical treatment of Ebstein anomaly in 45 adult patients
Chen LIU ; Mengping SHAO ; Tao HONG ; Wenjun DING ; Dong ZHAO ; Chunsheng WANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2012;(10):596-598
Objective To review retrospectively the experience of surgical treatment of Ebstein anomaly in 45 adult patients.Methods Between January 2008 and December 2011,45 adult patients underwent surgery for Ebstein anomaly.There were 14 male and 31 female patients aged 18 to 61 years (mean,39 years).The main symptoms included chest tightness,shortness of breath,palpitation.25 patients had cyanosis,while 6 patients with Wolff-Parkinson-White syndrome.Preoperative echocardiography: The septal downward from 2.5 cm to 7.5 cm,average 4.2 cm; the posterior leaflet downward 3.0 cm to 8.0cm,average 5.1 cm.Tricuspid incompetence was moderate in 6 patients and severe in 29.According to Carpentier classification,5 cases of type A,30 type B,type C in 6 cases.Preoperative New York Heart Association(NYHA) functional class Ⅱin 14 cases,Ⅲ 30 cases,Ⅳ in 1 cases.Surgical procedures included modified Danielson's in 15 patients,modified Carpentier's in 23,and 7 patients underwent tricuspid valve replacement(incloud 4 secondary operations).The ratio of tricuspid valve repair was 84.4%.Autologous pericardium patch was used to reconstruct the septal and posterior leaflets in 5 cases.Intraoperative use of annuloplasty rings in 21 cases.Results There were 2 hospital deaths with a postoperative mortality of 4.4%.The main causes of death were low cardiac output and right heart failure.Operation to discharge time was (8.0 ± 3.8)days.The follow up time was 2 to 46 months.Echocardiography examination: Within the 32 patients of severe tricuspid regurgitation before tricuspid valve plasty,16 patients’ tricuspid regurgitation disappeared,13 patients with mild regurgitation,3 moderate regurgitation.Within 5 patients of moderate tricuspid regurgitation before tricuspid valve plasty,4 cases tricuspid regurgitation disappeared after operation,1 case tricuspid valve mild regurgitation.Tricuspid valve leaflets were at normal levels,the atrialized portion of the ventricular disappeared.The heart function improved to be NYHA class Ⅰ to Ⅱ in 95 % of the survived patients.Conclusion Operative method of Ebstein anomaly sould be selected according to the type of pathoanatomies.Tricuspid valve repair should be performed to the best of our ability.