3.Damage of endothelial cells induced by OX-LDL and its influence in expressions of TLR-4 and EpCAM
Wei CHEN ; Kui ZHANG ; Chenhao ZHANG ; Qiang LI ; Yan LI
Journal of Jilin University(Medicine Edition) 2015;(6):1144-1149
Objective To study the damage effect of oxidized low density lipoprotein (OX-LDL)on the endothelial cells and its influence in the expressions of Toll like receptor 4 (TLR-4)and epithelial cell adhesion molecule (EpCAM).Methods The human endothelial cells ECV-304 were cultured in vitro and treated by different concentrations of OX-LDL for 24 h,and the cell vitality was measured by Taipan blue rejection test.The reactive oxygen species (ROS)level,mitochondrial membrane potential (MMP),cell cycle and apoptosis were detected by flow cytometry (FCM).The expression of TLR-4 and EpCAM were also analyzed by FCM.Results Compared with control group,the ROS levels in ECV-304 cells in 25,50,100,and 200 mg·L-1 OX-LDL were increased, but the cell vitalities and MMP were decreased (P < 0.05).Compared with control group,the percentages of S phase cells in 25 and 50 mg·L-1 OX-LDL groups were increased,the G0 G1 phase cells were slightly reduced,and the SubG1 phase cells (apoptotic cells)were increased with the increasing of OX-LDL concentration.Compared with control group,the TLR-4 and EpCAM expression levels in ECV-304 cells in 50 mg·L-1 OX-LDL group at 24 h were significantly increased (P < 0.05 ). Conclusion OX-LDL could increase the ROS level and induce the apoptosis in vascular endothelial cells,while increase the cell vitality and MMP.During the damage process,the expresion levels of TLR-4 and EpCAM are up-regulated.
4.Study on the effects of heat and ultraviolet fight radiation on hepatitis C virus JFH-1 strain inactivation
Hongshuo SONG ; Shuang SHI ; Ling YAN ; Kui LI ; Hui ZHUANG
Chinese Journal of Microbiology and Immunology 2009;29(5):438-442
Objective To investigate the inactivating effect of heat and ultraviolet(UV) light on HCV JFH-1 strain using the cell culture system. Methods The HCV JFH-1 virus stock, with an initial titer of 2.5 × 104 FFU/ml, was exposed in 56℃ water bath or to UV light for varying durations of time for explo-ring their inactivating effects on the virus. The kinetics of virus titer reduction was determined by an indirect immuno-fluorescence assay (IFA). If the cells infected with the exposed virus stock were IFA negative after three blind passages, the virus stock was considered to be inactivated completely. Results After incubation of the HCV JFH-1 virus stock (2.5 × 104 FFU/ml)in 56℃ water bath for 10 min, 20 min and 30 min, the virus titers were reduced to 1.6 × 103 FFU/ml, 3.1 × 102 FFU/ml and 3.3 × 10 FFU/ml, respectively. The exposure of the virus stock to UV light (wavelength 253.7 nm, intensity ≥60 μW/cm2, 30 cm below the UV lamp) for 15 s, 30 s and 45 s resulted in virus fiter reduction to 1.0 × 103 FFU/ml, 1.1 × 102 FFU/ml and 2.7 × 10 FFU/ml, respectively. After 40 min incubation of the virus stock at 56℃, or 1 min exposure to UV light (wavelength 253.7 nm, intensity ≥60 μW/cm2) the virus infectious titer was reduced below the detection limit of IFA, and the IFA was still negative even after three blind passages, indicating that the virus was inactivated completely. Conclusion HCV is sensitive to heat and UV light treatment. For HCV JFH-1 virus stock containing 2.5 × 104 FFU/ml virus, heat treatment at 56℃ for 40 min, or UV light expo-sure at an intensity of ≥60 μW/cm2 for 1 min, resulting in complete virus inactivation.
5.Relationships between high altitude de-adaptation syndrome and training burnout of plateau soldiers after returning to the plain
Kui DING ; Quanchao LI ; Yan WANG ; Xingzhen MENG ; Tian QIN
Military Medical Sciences 2016;40(8):661-664,694
Objective To explore the relationship between high altitude de-adaptation syndrome and training burnout of plateau-garrisoned soldiers after returning to the plain.Methods A total of 140 plateau-garrisoned soldiers who had returned to the plain were chosen by random cluster sampling and measured with the Training Burnout Test.They were divided into two groups according to the occurrence of acute mountain sickness.We compared the differences in training burnout between the two groups and analyzed the relationship between acute mountain sickness and training burnout. Results ①The incidence of high altitude de-adaptation syndrome of plateau-garrisoned soldiers was 80.00% after returning to the plain.There was statistically significant difference between plateau soldiers after returning to the plain,the plateau stability-keeping forces that returned to the plain (78.36%) (χ2 =0.188, P=0.664) and plateau migrants who returned to the plain (75.10%) (χ2 =1.279, P=0.258).Main symptoms of high altitude de-adaptation were fatigue (12.86%), dizziness (11.43%) and meakness (10.00%).②Compared with soldiers who had no high altitude de-adaptation syndrome, victims of de-adaptation syndrome had higher scores of training burnout, physical and psychological exhaustion and training-alienation (P<0.01).Compared with soldiers who had normal body mass indexes, those who were overweight had higher incidence of altitude de-adaptation (P<0.05).③The regression equation between plateau de-adaptation syndrome and training burnout of plateau soldiers after returning to the plain (P<0.01) is:Training burnout =0.498 ×plateau de-adaptation syndrome -0.126 ×age+0.038 ×length of military service +0.069 ×educational degree+0.029 ×body mass index.Conclusion There exist correlations between high altitude de-adaptation syndrome and training burnout of plateau soldiers after returning to the plain.Reducing the occurrence of high altitude de-adaptation syndrome can help reduce the degree of training burnout.
6.Asplenia syndrome complicated by dextrocardia and cerebral infarction: a case report.
Zhan-Kui LI ; Hua KE ; Jing LI ; Hai-Yan LIU ; Xiao-Peng LI ; Run-Min LI
Chinese Journal of Contemporary Pediatrics 2008;10(1):105-106
Cerebral Infarction
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etiology
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Dextrocardia
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etiology
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Female
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Humans
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Infant
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Spleen
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abnormalities
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Syndrome
7.Study on dsRNA Mediated Gene Silencing in Saccharomyces cerevisiae:Suppressing The Expression of GRE3 gene
Yan LI ; Ming YAN ; Kui WANG ; Lin XU ; Wei XU ; Pingkai OUYANG
Progress in Biochemistry and Biophysics 2007;34(3):292-298
RNA silencing is increasingly employed as an experimental strategy to probe for gene function in several organisms. The purpose of the present study was to test the effect of gene silencing by dsRNA in budding yeast Saccharomyces cerevisiae. GRE3 gene encoding an NADPH-dependent aldose reductase was chosen as an example. A recombinant plasmid psiLENT-GRE3 was constructed based on the pESC-LEU backbone and used to transform S. cerevisiae YPH499. The down regulation of GRE3 gene expression by inducing 1 kb RNA duplex and a 136 bp loop was investigated using reverse transcription- PCR. The results showed that double-stranded RNA mediated gene silencing could be used as a functional tool to decrease the expression level of a specific gene in S. cerevisiae, which would contribute to the understanding of RNA interference in budding yeast.
8.Effect and mechanism of methyl protodioscin in protecting cardiomyocytes against anoxia/reoxygenation injury.
Zong NING ; Yi-kui LI ; Yan ZHOU
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(4):407-409
OBJECTIVETo study the effect and mechanism of methyl protodioscin (MPD), an active ingredients of yamogenin, in protecting cardiomyocytes (CMC) against anoxia/reoxygenation (A/R) injury.
METHODSCultured CMCs of neonatal SD rats were randomly divided into three groups, cells in Group A were untreated normal cells, cells in Group B and C were made to injury CMC model by A/R, and only those in Group C were treated with MPD. Levels of ATPase activity and lactate dehydrogenase (LDH) in cell membrane of CMCs were determined. Besides, the mRNA expression of sodium-calcium exchanger (NCX) in MPD treated CMCs was detected.
RESULTSAs compared with Group B, the degree of CMC injury was significantly milder and the activities of Na+ -K+ -ATPase and Ca2+ -Mg2+ -ATPase were higher in Group C after cells were treated with MPD in concentration of 10 microg/mL and 50 microg/mL. The mRNA expression of NCX in CMCs was down-regulated after MPD treatment (P < 0.05).
CONCLUSIONMPD could maintain the low calcium internal environment in CMCs by way of protecting the membranous function of Na+ -pump and Ca2+ -pump, and influencing the Ca2+ transmembrane transportation in CMCs.
Animals ; Cell Hypoxia ; Cells, Cultured ; Diosgenin ; analogs & derivatives ; pharmacology ; Myocardial Reperfusion Injury ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; Oxygen ; adverse effects ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology
9.Effect of paeonol on blood pressure and blood flow in artery of spontaneously hypertensive rats and its mechanisms related on vasomotion.
Jin-yan ZHANG ; Le ZHAO ; Yi-kui LI ; Wei-liang WENG
China Journal of Chinese Materia Medica 2015;40(24):4903-4907
Previous studies have shown that paeonol can antagonize acute myocardial ischemia and infarction in rat. This study further researched the effects of paeonol on blood pressure and blood flow in the artery of spontaneously hypertensive rats and its mechanisms related on vasomotion. Firstly, thirty spontaneously hypertensive rats were randomly divided into spontaneously hypertensive control group and paeonol-treating groups of high dose and low dose, and also, the other ten Wistar rats as healthy control group. Before and after the intraduodenal administration of the drug, arterial blood pressure was measured by carotid artery and blood flow through the renal artery and carotid artery in vivo were measured by animal flowmeter. The same volume of solvent was given to the spontaneously hypertensive control group and the healthy control group, and the other operations were same. In order to further study the effect of paeonol on vasomotor function, the superior mesenteric artery, renal artery and coronary artery of the spontaneously hypertensive rat were removed and separated, precontracted by a certain concentration of potassium chloride (KCl) and 5-serotonin (5-HT) respectively, and dilatory responses were assessed by cumulative addition of paeonol. Results showed that after duodenal one-time delivery of paeonol, the blood pressure significantly lowered, the renal arterial blood flow and the carotid arterial blood flow significantly increased in spontaneously hypertensive rat. And also, paeonol relaxed the mesenteric artery, renal artery and the coronary artery of spontaneously hypertensive rat in a concentration-dependent manner. These results indicated that the effect of paeonol on decreasing arterial blood pressure and increasing the arterial blood flow was related to its vasodilative effect.
Acetophenones
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pharmacology
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Animals
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Blood Pressure
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drug effects
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Male
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Rats
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Rats, Inbred SHR
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Regional Blood Flow
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drug effects
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Vasodilator Agents
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pharmacology
10.Survival prediction of the Bolondi substaging model for patients with intermediate-stage hepatocellular carcinoma after hepatectomy
Wenxin WEI ; Zhengqing LEI ; Kui WANG ; Yong XIA ; Jun LI ; Zhenlin YAN ; Feng SHEN
Chinese Journal of Digestive Surgery 2016;15(5):496-503
Objective To investigate the overall survival prediction of the Bolondi substaging model for patients in intermediate-stage of Barcelona clinic liver cancer (BCLC) after hepatectomy.Methods The retrospective cohort study was adopted.The clinical data of 343 patients with intermediate-stage hepatocellular carcinoma (HCC) who were admitted to the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University between February 2008 and January 2010 were collected.All the patients received the detailed medical history collection,physical examination,laboratory and imaging examinations after admission,and then hepatectomy was performed according to the results of above examinations.Research methods:(1) patients were allocated into the B1,B2 and B3/4 groups based on the Bolondi's substaging model,and the prognostic analyses among groups were conducted.(2) The related factors affecting the prognosis of patients in the B1 and B2 groups were analyzed.(3) The patients in the B1 and B2 groups were allocated into the 4 groups [patients of B1 group with negative microvascular invasion (MVI) were divided in the M1 group,patients of B1 group with positive MVI in the M2 group,patients of B2 group with negative MVI in the M3 group and patients of B2 group with positive MVI in the M4 group] according to the situations of MVI,and stratified analysis was conducted.Observation indicators:basic clinical and pathological features and survival of patients in the B1,B2 and B3/4 groups were observed.Risk factors analysis affecting the prognosis of patients and stratified analysis of MVI in the B1 and B2 groups were conducted.All the patients were followed up by outpatient examination and telephone interview up to February 2014,and the abdominal ultrasound,liver function and serum alpha-fetoprotein (AFP) tests was performed once every 3 months within 2 years postoperatively and once every 6 months after 2 years postoperatively.The continuous variables and categorical variables were respectively represented as M(Qn) and percentage.The comparisons of continuous variables and categorical variables among groups were analyzed by ANOVA or Kruskal-Wallis test and chi-square test or Fisher exact probability,respectively,and one-way ordinal categorical variables were analyzed by the Kruskal-Wallis test.The survival curve was drawn using the KaplanMeier method.The univariate analysis and multivariate analysis were done using the Log-rank test and COX regression model.Results (1) The basic clinical pathological features:of 343 patients with HCC,143,183 and 17 patients (12 in the B3 substaging and 5 in the B4 substaging) were respectively allocated into the B1,B2 and B3/4 groups.There were statistically significant differences in the age,peritoneal effusion,total bilirubin (TBil),albumin (Alb),alanine transaminase (ALT),prothrombin time (PT),platelet (PLT),alpha-fetoprotein (AFP),extent of liver resection,surgical margin ivasion,tumor diameter,number of tumor,Edmondson-Steiner grade,Up-to-7 score,Up-to-7 standard and Child-pugh score among the 3 groups (F =3.377,NA,11.245,32.616,6.884,11.564,33.100,12.902,NA,NA,239.089,10.357,x2=8.906,F =251.508,x2 =343.000,106.790,P < 0.05).(2) Survival of patients:all the patients were followed up for 2.8-70.8 months with a median time of 38.7 months.The 1-,3-,5-year survival rates and median survival time in the B1,B2 and B3/4 groups were 85.8%,72.8%,52.9% and 63.2%,47.5%,16.8% and 45.5%,30.4%,8.4% and 55.1 months,35.1 months,12.2 months,respectively,showing a statistically significant difference (x2 =22.800,P < 0.05).(3) Risk factors analysis:the results of univariate analysis showed that the peritoneal effusion,Alb,Hb,AFP,esophagogastric varices,surgical margin invasion,tumor diameter,MVI and Edmondson-Steiner grade were related risk factors affecting the prognosis of patients with HCC after hepatectomy [HR =2.04,2.46,2.50,1.78,1.55,3.54,1.71,1.76,1.69,95% confidence interval (CI):1.13-3.69,1.20-5.02,1.51-4.15,1.29-2.45,1.06-2.25,1.65-7.61,1.23-2.38,1.23-2.51,1.08-2.64,P<0.05].The results of multivariate analysis showed that the Alb < 35 g/L,Alb < low limit of normal,tumor invading to surgical margin,tumor diameter > 5 cm and positive MVI were independent risk factors affecting the overall survival of patients with HCC after hepatectomy (HR =2.82,2.16,2.93,1.48,1.53,95% CI:1.37-5.80,1.27-3.69,1.33-6.44,1.05-2.09,1.06-2.22,P<0.05).(4) There were 61,82,57 and 126 patients in the M1,M2,M3 and M4 groups,and M2 and M3 groups were merged into the M2/3 group because of being similar survival situations of patients.The 1-,3-,5-year survival rates and median survival time in the M1,M2/3,and M4 groups were 90.0%,83.2%,67.7% and 68.8%,59.9%,41.6% and 52.7%,42.1%,23.6% and 69.0 months,49.2 months,24.9 months,respectively,with a statistically significant difference among the 3 groups(x2=20.200,P < 0.05).Conclusions The Bolondi substaging model produces an optimal survival prediction for patients in intermediate stage of BCLC after hepatectomy.The patients in the B1 and B2 substaging have better long-term survival outcomes after hepatectomy.