3.Effects of Akt signal pathway on the expression of NF-κB in renal tubular epithelial cells
Ling YANG ; Ping GAO ; Xiaoyun SI ; Ling CHEN ; Hua SHUI
Chinese Journal of Emergency Medicine 2011;20(2):173-176
Objective To investigate the role of Akt signal pathway on the expression of monocyte chemoattractant protein-1 ( MCP-1 ) and nuclear transcription factor-κB (NF-κB) in renal tubular epithelial cells (HK-2) stimulated by albumin and to explore the mechanisms of action. Method The HK-2 cells were incubated in the presence of albumin (5,15,30 mg/mL) with or without Ly294002 (an inhibitor of Akt). Expression of mRNA was analyzed by reverse transcription-polymerase chain reaction (RT-PCR).Expression of Akt and protein MCP-1 were assessed by Western blot. Electrophoretic mobility shift assay (EMSA) was used to detect the activation of NF-κB. q-test was used to evaluate the differences in means between groups. Results Compared with control group, the expression of MCP-1 mRNA remarkly increased. [Control group: 0.233 ±0.01; BSA(5 mg/mL) group: 0.285 ±0.04; BSA( 15 mg/mL) group:0.387 ± 0.02; BSA ( 30 mg/mL) group: 0.473 ± 0.05; BSA ( 30 mg/mL) + Ly294002 group: 0. 325 ±0.05, P < 0.05 ]. The expression of MCP-1 protein in renal interstitum of operation group were remarkly increased too. [ Control group: 100 ± 15.1; BSA ( 5 mg/mL) group: 148 ± 19.3; BSA ( 15 mg/mL) group: 176±20.7; BSA(30 mg/mL) group: 263 ± 18.1; BSA(30 mg/mL) + Ly294002 group: 175 ± 18.0, P <0.05 ]. Albumin stimulated the expression of MCP-1mRNA and protein in a dose-dependent manner. Albumin remarkably increased the activity of NF-κB. Albumin enhanced the expression of Akt. Ly294002 inhibited albumin-induced the expression of NF-κB and partially decreased the level of MCP-1. Apositive correlation was noted between NF-κB activation and MCP-1 expression( r = 0.68 ,P < 0.01 ). Conclusions Albumin-induces MCP-1 and NF-κB production via Akt signal pathway in renal tubular epithelial cells.
5.Evaluation of the MICROTEST 1 ESR analyzer and investigation of the reference value
Li-Ya LI ; Wei-Bin CHEN ; Feng GAO ; Shui-Fen SHEN ; Hui-Ping JIN ;
Chinese Journal of Laboratory Medicine 2001;0(03):-
0.37).Meanwhile a good correlation (Y=0.99X-0.18,r=0.987) was obtained. Though Westergren method correlated preferably with MICROTEST 1 (Y=0.86X+1.27,r=0.906),there was a markedly different (t=3.174,P=0.001).At last different references values were collected, according to sex and age.Male,32.5 mm/1 h(60 years old);Female, 34.03 mm/1 h(50 years old).Conclusions MICROTEST 1 correlated preferably with Westergren method.The examination by MICROTEST 1 needs small quantity of sample and fewer time.Furthermore,it has good repeatability and stability.The factors such as temperature and Hct have little influence on the results.The result suggested that it is suitable to apply MICROTEST 1 to large- scale clinical laboratory or other labs.But the reference value of ESR was influenced by age,which should be considered in clinical usage.
6.Improvement of Methods of Making Myocardial Ischemia-Reperfusion Model and Effects on Electrocardiogram and Ultrastructure of Myocardium in Rats
yan-wei, LIU ; shui-ping, LIU ; jian-ding, CHENG ; yu-chuan, CHEN
Journal of Applied Clinical Pediatrics 2004;0(07):-
Objective To improve the methods of making myocardial ischemia-reperfusion model and observe the changes of electrocardiogram and ultrastructure of myocardium in rats.Methods The chest of young male SD rats through the fourth intercostal space was opened,and the left coronary artery was tied with a silicagel tube,after 30 minutes,untied to perfuse.Changes of electrocardiogram were observed and recorded.After reperfusion,the levels of AST,LDH,and CK-MB were measured and the tissue samples of the infarct areas were examined by transmission electronic microscope.Results The 90 percent of total rats were made myocardial ischemia reperfusion model successfully.In myocardial ischemia reperfusion rats:the QRS wave of myocardial ischemia-reperfusion rats was much higher than that of control group;the level of cardiac enzymes increased;myocardial and vascular endothelial cells ultrastructure was damaged seriously.Conclusions The improvement of modus operandi is right.Ischemia reperfusion can cause evident damage of myocardial and vascular endothelial cells ultrastructure in rats,and damage of myocardial cells is more severe than vascular endothelial cells.
7.Correlation between Qualities of Life and Depression,Anxiety for Cancer Patients
Li-ping GAO ; Chang-Shui WENG ; Hong ZHAO ; Liu YANG ; Ying LIU ; Lijun YUAN ; Dianxuan CHEN ; Lei WANG
Chinese Journal of Rehabilitation Theory and Practice 2006;12(3):192-193
ObjectiveTo determine the correlation between qualities of life(QOL) and depression,anxiety in cancer patients.Methods91 cancer patients was measured with self anxiety scale(SAS),self depression scale(SDS) and SF-36.ResultsDepression occurred in about 51.5% and anxiety occurred in about 24.1% of the general patients.The patients without depression showed a higher QOL than those with depression(P<0.05),and the patients without anxiety showed a higher QOL than those with anxiety(P<0.01).ConclusionDepression and anxiety are associated with poor quality of life,which should be put into plans of nursing.
8.The changes of electrocardiogram and serum cardiac troponin I at the early stage of crush injury in rats.
Yu-chuan CHEN ; Shui-ping LIU ; Wei GUO
Journal of Forensic Medicine 2002;18(2):76-77
OBJECTIVE:
To observe the changes of electrocardiogram and serum cardiac troponin I at the early stage of severe crush injury in rats.
METHODS:
Crush injury was produced in Sprague-Dawley rats. The changes of electrocardiogram were recorded with the standard II, the serum levels of cardiac troponin I were studied by automated chemiluminescence assay.
RESULTS:
The ST segment elevated considerably after crush injury and lasted 24 h, the levels of serum cTnI were much higher than those of the control groupes after 6 h of injury.
CONCLUSION
Cardiomyocyte injury was induced in the early phase of crush injury.
Animals
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Crush Syndrome/physiopathology*
;
Electrocardiography
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Extremities/injuries*
;
Female
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Heart Injuries/physiopathology*
;
Male
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Rats
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Rats, Sprague-Dawley
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Troponin I/blood*
9.Study on steroidal saponins from Dioscorea zingiberensis and their platelet aggregation activities.
Jing-jing WANG ; Yi-xun LIU ; Di WEN ; He-shui YU ; Li-ping KANG ; Xu PANG ; Zhao YANG ; Bai-ping MA ; Yun-dai CHEN
China Journal of Chinese Materia Medica 2014;39(19):3782-3787
Using the absorbent resin, silica gel and ODS column chromatography as well as semi-preparative HPLC, ten compounds were isolated from 70% ethanol extract of tubers of Dioscorea zingiberensis C. H. Wright, and their structures were elucidated as trigoneoside XIIIa (1), parvifloside (2), trigoneoside IVa (3), deltoside (4), protobioside (5), lilioglycoside k (6), zingiberensis newsaponin I (7), deltonin (8), prosapogenin A of dioscin (9), and trillin (10) on the basis of NMR and MS spectral data analysis. Among these compounds, 1, 3, 5 and 6 were isolated from this plant for the first time. In the screening test on platelet aggregation, compounds 7 and 8 exhibited induction effect on platelet aggregation, while compound 9 exhibited significant inhibitory effect on platelet aggregation in vitro.
Animals
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Dioscorea
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chemistry
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Drugs, Chinese Herbal
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chemistry
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pharmacology
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Male
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Mass Spectrometry
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Molecular Structure
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Platelet Aggregation
;
drug effects
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Rats
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Rats, Wistar
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Saponins
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chemistry
;
pharmacology
10.Detection of enterohemorrhagic Escherichia coli O157:H7 by loop-mediated isothermal amplification.
Shui-rong ZHU ; Yin CHEN ; Zhi-gang WANG ; Da-zhi JIN ; Qun-ying LU ; Ping-ping YAO
Chinese Journal of Preventive Medicine 2009;43(9):803-808
OBJECTIVETo develop a loop-mediated isothermal amplification (LAMP) method for rapidly diagnosing of Escherichia coli (EHEC) O157:H7 in pathogen detection department or small-scale laboratories.
METHODSPrimers for LAMP test were designed by targeting the antigen coding rfbE of EHEC O157:H7, the Shiga-like toxin stx2 and the fliC encoding gene of H7 flagella antigen, respectively. The reaction condition and reaction system of LAMP were optimized. 2 EHEC O157:H7 type strains, 17 local strains and 33 other enterobacteria were analyzed to evaluate the LAMP's specificity and sensitivity. The results of the LAMP reaction were also compared with routine PCR method.
RESULTSThe amplification products of O157 which had the corresponding target genes turned green by visual inspection and had ladder-like pattern on the gel, but products of other enterobacteria remained orange by visual examination and had no band on the gel. The detection results of LAMP were the same as of routine PCR method. The reaction time of the LAMP method was only 1.5 hours and the detection limit of LAMP assay was 26 CFU/reaction. In addition, the LAMP results could be determined only by visual inspection.
CONCLUSIONLAMP assay is rapid, specific, and sensitive for the detection of EHEC O157:H7. This method might not only reduce the dependence of complicated equipments but also be a potential method for wider use in pathogen detection department, small-scale laboratory, emergency motor vehicle or field survey.
Environmental Monitoring ; methods ; Escherichia coli O157 ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods ; Sensitivity and Specificity