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1.
Braz. j. med. biol. res ; 54(2): e9017, 2021. graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS | ID: biblio-1142574

ABSTRACT

The purpose of this study was to investigate the anti-cancer effect of melittin on growth, migration, invasion, and apoptosis of non-small-cell lung cancer (NSCLC) cells. This study also explored the potential anti-cancer mechanism of melittin in NSCLC cells. The results demonstrated that melittin suppressed growth, migration, and invasion, and induced apoptosis of NSCLC cells in vitro. Melittin increased pro-apoptotic caspase-3 and Apaf-1 gene expression. Melittin inhibited tumor growth factor (TGF)-β expression and phosphorylated ERK/total ERK (pERK/tERK) in NSCLC cells. However, TGF-β overexpression (pTGF-β) abolished melittin-decreased TGF-β expression and pERK/tERK in NSCLC cells. Treatment with melittin suppressed tumor growth and prolonged mouse survival during the 120-day observation in vivo. Treatment with melittin increased TUNEL-positive cells and decreased expression levels of TGF-β and ERK in tumor tissue compared to the control group. In conclusion, the findings of this study indicated that melittin inhibited growth, migration, and invasion, and induced apoptosis of NSCLC cells through down-regulation of TGF-β-mediated ERK signaling pathway, suggesting melittin may be a promising anti-cancer agent for NSCLC therapy.


Subject(s)
Animals , Rabbits , Apoptosis , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/drug therapy , MAP Kinase Signaling System , Lung Neoplasms/genetics , Lung Neoplasms/drug therapy , Melitten/pharmacology , Down-Regulation , Gene Expression Regulation, Neoplastic , Cell Movement , Transforming Growth Factor beta/metabolism , Cell Line, Tumor , Caspase 3 , Apoptotic Protease-Activating Factor 1 , Neoplasm Invasiveness
2.
Chinese Journal of School Health ; (12): 728-732, 2021.
Article in Chinese | WPRIM | ID: wpr-877139

ABSTRACT

Objective@#To provide a basis for college students mental health education and guidance, the impact of psychological aggression and corporal punishment college students experienced before the age of 10 on anxiety and health risk behaviors was explored.@*Methods@#In April and May 2019, the convenience sampling method was used to conduct a questionnaire survey among all the freshman and sophomore students of some majors in two universities in Harbin, including a total of 2 150 students. The contents of the questionnaire included general information survey, Parent-Child Conflict Tactics Scale (CTSPC),Self-Rating Anxiety Scale (SAS) and Adolescent Health related Risky Behavior Inventory (AHRBI).@*Results@#The percentage of college students who suffered psychological aggression and corporal punishment before the age of 10 was 80.05% and 58.37%. The detection rate of corporal punishment in boys and non-only child was significantly higher than that in girls and only child (P<0.05). The students who experienced psychological aggression and corporal punishment before the age of 10 had higher detection rates of anxiety state and six subscales of health risk behaviors than those who didn t experienced those (P<0.05). The Logistic regression analysis showed that, psychological aggression, place of residence, corporal punishment, and only child were positively associated with college students anxiety state (OR=1.85,1.53,1.49,1.34). The Logistic regression analysis showed that, psychological aggression, place of residence were positively associated with college students health-compromising behaviors (OR=1.60,1.26). Psychological aggression, gender factors, corporal punishment, and only child all influenced college students aggression and violence behavior (OR=1.75,1.66,1.63,1.28). Corporal punishment influenced college students suicide and self-injury behavior (OR=3.44). Gender factors and corporal punishment all influenced college students smoking and drinking behavior (OR=2.67,2.25).Corporal punishment and psychological aggression were positively associated with college students rule breaking behavior (OR=1.88,1.55). Corporal punishment, gender factors, and family income all influenced college students unprotected sexual behavior (OR=4.51,1.91,0.62).@*Conclusion@#Parental psychological aggression and corporal punishment college students experienced during childhood have a negative impact on their anxiety and health risk behaviors.

3.
Chinese Journal of Geriatrics ; (12): 1020-1024, 2020.
Article in Chinese | WPRIM | ID: wpr-869518

ABSTRACT

Objective:To analyze risk factors for deep vein thrombosis(DVT)in patients with severe cerebral infarction and to find early and sensitive indicators for the prediction and intervention of DVT.Methods:A total of 226 patients with severe cerebral infarction aged 62.5±12.9 years in our department from January 2017 to May 2020 were enrolled.Clinical data, biochemical examinations and color Doppler ultrasound results were collected.Risk factors for DVT were analyzed.The receiver operating characteristic curve(ROC)was used to determine the cut-off value, area under the curve, sensitivity and specificity.Results:Age, reaction(R)time of blood coagulation factors on thromboelastography(TEG)and fibrinogen degradation products(FDP)were risk factors for DVT with no adjustment of the overall effect of time on coagulation mechanisms.According to time stratified analysis, decreased R time( OR=0.58, 95% CI: 0.40-0.84)and increased FDP( OR=1.17, 95% CI: 1.02-1.33)within 3 days of onset were risk factors for DVT, and the cut-off values were 5.35 min and 0.39 mg/L, respectively; 3 and 7 days after onset, increased D-dimer was a risk factor for DVT( OR=2.73, 95% CI: 1.53-4.86; OR=2.57, 95% CI: 1.32-5.03), and the cut-off values were 0.39 mg/L and 0.76 mg/L, respectively.Excluding the effects of FDP primary and D-Dimer secondary fibrinolysis, risk factors for DVT within 3 days of onset were decreased R time on TEG and increased age, and all risk factors were not statistically significant 3 days and 7 days after onset( P<0.05). Conclusions:The key factors affecting DVT in patients with severe cerebral infarction are different at different stages.Decreased R time within 3 days of onset is a predictive indicator of DVT.FDP and D-dimer can be used to assess thrombosis, but may not be appropriate as predictive indicators.

4.
Article in Chinese | WPRIM | ID: wpr-815605

ABSTRACT

@# Objective: To study the expression of miR-142-5p in lung adenocarcinoma tissues, and to explore its effect on proliferation, invasion, migration and epithelieal-mesenchymal transition (EMT) of H1650 cells and the potential mechanisms. Methods:Atotal of 107 pairs of lung adenocarcinoma tissues and corresponding para-cancerous tissues from patients, who underwent tumor resection and were pathologically confirmed at the Department of Thoracic Surgery, the Fourth Hospital of Hebei Medical University between Jan. 2014 and Jan. 2015, were collected for this study; in addition, human lung adenocarcinoma cell lines (H1650, HCC827, A549, H1975, PC9) and human bronchial epithelial BEAS-2B cells were also used in this study. qPCR was used to detect the expression of miR-142-5p in lung adenocarcinoma tissues and cell lines. The correlation between expression of miR-142-5p and clinical features was analyzed.After transfection with miR-142-5p mimics or miR-negative control (miR-NC) plasmid, the proliferation, invasion and migration of H1650 cells were detected with CCK-8, Transwell invasion assay and Wound healing assay, respectively. The bioinforamtics tool was used to predict the target genes of miR-142-5p, and Luciferase reporter gene assay was performed to validate the regulation of miR-142-5p on target gene. Western blotting (WB) was used to detect the expressions of cyclin-dependent kinase 5 (CDK5) and EMTrelated protein. Results: Compared to Para-cancerous tissues and BEAS-2B cells, the expression of miR-142-5p was lower in lung adenocarcinoma tissues and cell lines (all P<0.01). Of the 107 cases of lung adenocarcinoma tissues, 61 cases (57.01%) showed decreased miR-142-5 expression, which was correlated with the TNM stage and lymph node metastasis (both P<0.01). Transfection of miR-142-5p mimics significantly up-regulated the expression of miR-142-5p and decreased the proliferation, invasion and migration of H1650 cells (all P<0.05 or P<0.01). Bioinformatics showed that CDK5 was a target gene of miR-142-5p. Luciferase reporter gene assay and WB validated that miR-142-5p could significantly down-regulate CDK5 expression in H1650 cells, up-regulate the expression of E-cadherin and down-regulate the expressions of N-cadherin, Twist and Snail in H1650 cells (all P<0.01). Conclusion: miR-142-5p is low expressed in lung adenocarcinoma tissues and cell lines; it suppresses the EMT process to inhibit, invasion and migration of H1650 cells via down-regulating the expression of CDK5.

5.
Chinese Journal of School Health ; (12): 1339-1342, 2020.
Article in Chinese | WPRIM | ID: wpr-829122

ABSTRACT

Objective@#To compare life satisfaction and social desirability between only and non-only children in urban and rural areas, and to provide a basis for future mental health education guidance.@*Methods@#Stratified cluster sampling method was used to select 3 765 students of grade 4 and 5 from 9 primary schools in urban and rural area of Harbin. The contents of the questionnaires included general information survey, the Multidimensional Student’s Life Satisfaction Scale (MSLSS) and the Children’s Social Desirability Scale (CSD).@*Results@#In the overall life satisfaction, county children had the highest satisfaction (5.10±0.50), while rural children had the lowest satisfaction (4.96±0.57);The life satisfaction of girls was higher than that of boys; The life satisfaction of only children was higher than that of non-only children(F/t=17.04,-5.91,2.21,P<0.05). In terms of social desirability, the social desirability of urban children(32.33±7.04) and county children (31.78±6.30) was higher than that of rural children(30.82±6.34).The social desirability of girls was higher than that of boys, the only children was higher than that of the non-only children (P<0.05). In the overall life satisfaction and all domains of life satisfaction apart from friend life satisfaction, the life satisfaction of urban girls was higher than that of boys(P<0.05). Except for county non-only children, the social desirability of girls was higher than that of boys (P<0.05). In the overall life satisfaction and all domains of life satisfaction apart from family life satisfaction, the life satisfaction of county boys was highest among three areas (P<0.05). In all domains of life satisfaction, the life satisfaction of county girls was highest and the life satisfaction of rural girls was lowest(P<0.05). In the family life satisfaction, the life satisfaction of only boys in urban areas was higher than that of non-only boys(P=0.03). In the overall, friend, living environment and self-dimension life satisfaction, the only and non-only children in the urban, county and rural area had significant differences (P<0.05). In addition, the life satisfaction of county children was highest and the life satisfaction of rural children was lowest. The social desirability of only children in urban and county areas was higher than that of only children in rural areas.@*Conclusion@#There are differences in life satisfaction and social expectations among urban, county and rural children in Harbin. Corresponding intervention strategies should be developed when administers psychological counselling to children.

6.
Neuroscience Bulletin ; (6): 47-56, 2019.
Article in English | WPRIM | ID: wpr-775463

ABSTRACT

Angiotensin (Ang)-(1-7) is an important biologically-active peptide of the renin-angiotensin system. This study was designed to determine whether inhibition of Ang-(1-7) in the hypothalamic paraventricular nucleus (PVN) attenuates sympathetic activity and elevates blood pressure by modulating pro-inflammatory cytokines (PICs) and oxidative stress in the PVN in salt-induced hypertension. Rats were fed either a high-salt (8% NaCl) or a normal salt diet (0.3% NaCl) for 10 weeks, followed by bilateral microinjections of the Ang-(1-7) antagonist A-779 or vehicle into the PVN. We found that the mean arterial pressure (MAP), renal sympathetic nerve activity (RSNA), and plasma norepinephrine (NE) were significantly increased in salt-induced hypertensive rats. The high-salt diet also resulted in higher levels of the PICs interleukin-6, interleukin-1beta, tumor necrosis factor alpha, and monocyte chemotactic protein-1, as well as higher gp91 expression and superoxide production in the PVN. Microinjection of A-779 (3 nmol/50 nL) into the bilateral PVN of hypertensive rats not only attenuated MAP, RSNA, and NE, but also decreased the PICs and oxidative stress in the PVN. These results suggest that the increased MAP and sympathetic activity in salt-induced hypertension can be suppressed by blockade of endogenous Ang-(1-7) in the PVN, through modulation of PICs and oxidative stress.


Subject(s)
Angiotensin I , Metabolism , Animals , Antioxidants , Pharmacology , Blood Pressure , Hypertension , Drug Therapy , Male , Oxidative Stress , Paraventricular Hypothalamic Nucleus , Peptide Fragments , Metabolism , Rats, Sprague-Dawley , Reactive Oxygen Species , Metabolism , Sodium Chloride, Dietary , Pharmacology
7.
Chinese Journal of School Health ; (12): 236-238, 2019.
Article in Chinese | WPRIM | ID: wpr-819317

ABSTRACT

Objective@#To understand impacts of past experiences of school bullying victimization and social supports on anxiety symptoms of college students, and to provide basic data for appropriate mental health education.@*Methods@#By using cluster random sampling method, a questionnaire survey was conducted among freshman, sophomore and junior college students majoring in clinical medicine from one medical university in Heilongjiang Province. The questionnaire included general demographic information, school bullying and self-rating anxiety scale.@*Results@#In this survey, 647 students (28.4%) were reported of anxiety symptoms, including 462 (20.3%) with mild anxiety, 162 (7.1%) with moderate anxiety, and 23(1.0%) with severe anxiety. Reported rate of mild, moderate and severe anxiety among college students with bullying victimization experiences during primary school students was 22.8%,33.9% and 59.6%, respectively (χ2=93.05, P<0.05). The figure among students with bullying victimization during junior school was 21.5%, 45.2% and 57.3%, respectively (χ2=186.79, P<0.05). Among those with bullying victimization experiences, anxiety symptoms showed no differences between students with or without social supports from peers, parents and teachers (χ2=7.95, P=0.54; χ2=11.57, P=0.24). Ordinal logistic regression showed bullying victimization during primary school and affection to primary school positively associated with anxiety symptoms among college students, while bullying victimization experiences during junior school showed no association with anxiety symptoms in college.@*Conclusion@#Bullying victimization experiences during primary school shows significant association with anxiety symptoms among college students.

8.
Chinese Journal of School Health ; (12): 700-703, 2019.
Article in Chinese | WPRIM | ID: wpr-818697

ABSTRACT

Objective@#To compare behavioral and emotional health among first-born children and the only-child in Harbin, as well as associated factors including parents, family background, parenting and family environment.@*Methods@#A questionnaire survey was conducted for 156 parents of first-born children and the only-children, matched in age (<3 months), class and gender. Achenbach Child behavior scale (CBCL), the Self-evaluation of Anxiety Scale (SAS), the Self-rating Depression Scale(SDS), the Parents Rearing Behavior Questionnaire (CRPR) and the Family Assessment Device Scale (FAD) were used.@*Results@#There was no statistically significant difference in the scores of each dimension of children's emotional and behavioral health between the two groups(17.88±5.93)(19.13±6.01),total score(t=-0.74,P>0.05). There was no statistical difference in anxiety and depression between the two groups of parents(χ2=0.51,0.40,P>0.05); In terms of parenting style, the acceptance and encouragement achievement score for first-born children was significantly higher than that of the only child (t=2.10,2.12, P<0.05). In terms of family functions, there was no statistical difference in total function (t=-0.43, P>0.05). Behavioral problems associated with parents' anxiety, depression, parental rearing style and family function. Regression analysis showed that behavioral problems were mainly affected by sibling relationship for first-born children(B=8.74) and family role function for the only child (B=1.27).@*Conclusion@#No significant differences in behavioral and emotional health between first-born child and the only child are observed. However, harmonious sibling relationship, emotionally supportive parents and home environment could help improving behavioral and emotional health.

9.
Cancer Research and Clinic ; (6): 385-388, 2013.
Article in Chinese | WPRIM | ID: wpr-436628

ABSTRACT

Objective To observe the influence of p53-upregulated modulator of apoptosis (PUMA) on the growth of human brain glioma cell lines U251 (p53 mutant) and SHG-44 (p53 wild type),and to explore its possible mechanism.Methods Construct the adenovirus PUMA (Ad-PUMA) and vector of adenovirus (AdDsRed) which were respectively transfected into glioma cell lines U251 and SHG-44.Cells proliferation rates were measured with cell counting kit-8 (CCK-8).The apoptotic ratios were detected by flow cytometry.The expression of PUMA and apoptosis associated proteins (bcl-2,Bax) were determined with Western blot analysis.Caspase-3,Caspase-8,Caspase-9 activity were measured by Caspase activity assay kit.Results Compared with vector group and blank control group,Ad-PUMA transfected group showed strong cell proliferating inhibition effects [the inhibition rates were (50.89±4.73) % and (44.45±5.33) % respectively,P <0.05] and pro-apoptotic effects [apoptotic rates were (44.89±5.08) % and (31.67±7.32) %,P < 0.05] in different p53 glioma cell lines U251 and SHG-44.Western blot analysis showed that PUMA protein expression increased after Ad-PUMA transfection,accompanied by the reduced expression of the anti-apoptotic protein bcl-2 and the increased expression of pro-apoptotic protein Bax.The activity of Caspase testing results showed that the Caspase-3,Caspase-9 activity increased significantly,while the Caspase-8 activity changed little.Conclusion No matter how p53 phenotype,PUMA can inhibit glioma proliferation,promote apoptosis,and its mechanism may be through the mitochondrial apoptotic pathways,upregulation of Bax and inhibition of bcl-2 expression,which activated Caspase-9.Ad-PUMA is expected to become a new target for gene therapy of gliomas.

10.
Cancer Research and Clinic ; (6): 252-255, 2012.
Article in Chinese | WPRIM | ID: wpr-428663

ABSTRACT

Objective To explore differences of drug resistance of temozolomide(TMZ) on different CD133 immune prototype of glioma cells and study on the changes of their sensitivity to TMZ through increased PUMA. Methods CD133+-U87MG cells sorted by CD133 magnetic beads were cultured in serum-free stem cell medium respectively. The cells were infected with recombinant adenovirus, Ad-PUMA, diluted in cell culture medium with or without TMZ intervention.The inhibitory rate of cell proliferation was detected by MTT assay and 50 % inhibition concentration of TMZ was calculated. Apoptosis rates of CD133+-U87MG cells were assessed by flow cytometry (FCM) before and after intervention of exogenous PUMA and TMZ.Results The TMZ IC50 values of CD133+glioma cells were higher than that of CD133- glioma cells. There were significant differences in apoptosis rate between CD133+ glioma cell and CD133- glioma cell (all P<0.05).Conclusion AdPUMA joint TMZ can promote glioma stem cells apoptosis, thus improve the sensitivity to chemotherapy of glioma.

11.
Article in Chinese | WPRIM | ID: wpr-419110

ABSTRACT

ObjectiveTo investigate the mechanisms and the effects of magnesium Valproate on the expressions of the kinin B1 and B2 receptors in the hippocampus of the juvenile rats submitted to pilocarpine model of epilepsy.Methods 35 healthy Wistar juvenile rats were randomly divided into six groups,that is the model groups:Ⅰ group,Ⅱ group,Ⅲ group,and intraperitoneal injection of saline water control groups:Ⅰ a group,Ⅱ a group,Ⅲ a group,after succession of 15 rats to kindle to establish the model of epilepsy by pilocarpine.To collect hippocampus tissue after the rats were to put to death,and to compared the expression levels of kinin B1 and B2 receptor mRNA by RT-PCR and western blot in the hippocampus of rats.ResultsBy treated with magnesium valproate,kinin B1 receptor mRNA (0.38 ± 0.051 ) and protein expressions(0.58 ± 0.057 ) decreased and kinin B2 receptor mRNA (0.48 ±0.056 ) and protein expressions(0.48 ± 0.044 ) increased in Ⅰ group,compared with that (0.76 ±0.068,0.89 ± 0.034;0.28 ± 0.034,0.32 ± 0.039 ) of Ⅰ a group(P < 0.05 ).Compared with control group,there were more significant upregulation of kinin B1 receptor mRNA and protein expressions (P<0.05) in the Ⅰ and the Ⅱ groups and there were no alteration in Ⅲ group.The expressional levels of B2 receptor mRNA and protein were upregulated in the Ⅰ,Ⅱ and Ⅲ groups.ConclusionThe kinin B1 and B2 receptor may play a role in the onset and maintenance of epilepsy.The magnesium valproate increased the expressional levels of kinin B2 receptor,and decreased the expressional levels of kinin B1 receptor.

12.
Chinese Journal of Geriatrics ; (12): 525-528, 2012.
Article in Chinese | WPRIM | ID: wpr-426571

ABSTRACT

Objective To explore effects of atorvastatin on the expressions of cyclooxygenase-2(COX-2) and membrane-associated prostaglandin E2 synthase-1 (mPGES-1) in the carotid atherosclerotic plaques of rabbits.Methods Totally 33 male New Zealand white rabbits(≥ 36months of age ) were assigned into normal control group (n=8) and animal model group with carotid atherosclerotic stenosis (n =25).The rabbit models were randomly divided into non-intervention group,celecoxib treatment group (15 mg · kg-1 · d-1,twice daily) and atorvastatin treatment group (5 mg · kg-1 · d-1,once daily) (n=8 each).Four weeks after treatment,the mRNA and protein expressions of COX-2 and mPGES-1 in carotid plaques were determined by RT-PCR and Western blot,respectively.Results The mRNA expressions of COX-2 (0.97±0.09,0.44±0.05,0.60±0.04vs.0.23±0.04,F=66.77,P<0.01) and mPGES-1 (0.92±0.07,0.41±0.04,0.61±0.03 vs.0.17±0.03,F=54.87,P<0.01)in carotid atherosclerotic plaques were significantly higher in non intervention group,celecoxib treatment group and atorvastatin treatment group than in normal control group.The mRNA expressions of COX-2 and mPGES-1 were decreased in celecoxib treatment group and atorvastatin treatment group as compared with non-intervention group ( both P < 0.01 ).The protein expressions of COX-2 (0.89±0.06,0.42±0.07,0.62±0.04 vs.0.18±0.05,F=61.75,P <0.01) and mPGES-1(0.91±0.05,0.44±0.05,0.63±0.05 vs.0.21±0.04,F=86.44,P<0.01)in carotid atherosclerotic plaques in non-intervention group,celecoxib treatment group and atorvastatin treatment group were increased as compared with those in normal control group.The mRNA and protein expressions of COX-2 and mPGES-1 were decreased in celecoxib treatment group and atorvastatin treatment group as compared with non-intervention group(all P<0.01 ).The expressions of COX-2 and mPGES-1 in carotid atherosclerotic plaques were reduced in celecoxib treatment group as compared with atorvastatin treatment group (P < 0.01).Conclusions As COX-2 inhibitor celecoxib,atorvastatin may inhibit the expressions of COX-2 and mPGES-1,and interfere with the inflammatory response which plays key role in the pathological progress of carotid atherosclerotic plaques,and thus slow the progress of carotid atherosclerosis.

13.
Article in Chinese | WPRIM | ID: wpr-233174

ABSTRACT

The effect of Smac gene on the TRAIL-induced apoptosis of the prostate cancer cell line PC-3 and the molecular mechanism were investigated.The Smac gene was transfected into PC-3 cells under the induction of liposome.The intrinsic Smac gene expression was detected by Western blotting.After treatment with TRAIL as an apoptosis inducer,in vitro cell growth activity was assayed by MTT colorimetry.The apoptosis rate of PC-3 cells was determined by annexin V -FITC and propidium iodide staining flow cytometry.The expression of cellular XIAP and caspase-3 genes was examined by Western blotting.Smac-transfected cells (PC-3/Smac group) had significantly increased Smac protein level as compared with PC-3 controls (P<0.01).After induction with 100-200 ng/mL TRAIL for 12-36 h,cellular proliferation rate in PC-3/Smac group was significantly lower than in PC-3 controls (P<0.05).After induction with 100 ng/mL TRAIL for 24 h,the apoptosis rate in PC-3/Smac group was significantly enhanced as compared with that of PC-3 controls (P<0.05).Accordingly,the XIAP expression level was down-regulated significantly (P<0.05) and caspase-3 subunit P20 was up-regulated significantly (P<0.05).It is suggested that the over-expression of cellular Smac can inhibit inhibitor of apoptosis proteins (IAPs),enhance caspases activity and the apoptosis rate of PC-3 cells induced by TRAIL,which may provide a useful experimental basis for prostate cancer therapy.

14.
Article in Chinese | WPRIM | ID: wpr-233163

ABSTRACT

Many researchers employed mammalian expression system to artificially express cannabinoid receptors,but immunoblot data that directly prove efficient protein expression can hardly be seen in related research reports.In present study,we demonstrated cannabinoid receptor protein was not able to be properly expressed with routine mammalian expression system.This inefficient expression was rescued by endowing an exogenous signal peptide ahead of cannabinoid receptor peptide.In addition,the artificially synthesized cannabinoid receptor was found to aggregate under routine sample denaturing temperatures (i.e.,≥95℃),forming a large molecular weight band when analyzed by immuno-blotting.Only denaturing temperatures ≤75℃ yielded a clear band at the predicted molecular weight.Collectively,we showed that efficient mammalian expression of cannabinoid receptors need a signal peptide sequence,and described the requirement for a low sample denaturing temperature in immuno-blot analysis.These findings provide very useful information for efficient mammalian expression and immuno-blotting of mcmbrane receptors.

15.
Cancer Research and Clinic ; (6): 150-153, 2011.
Article in Chinese | WPRIM | ID: wpr-413261

ABSTRACT

Objective To investigate the inhibitive effects of Ad-PUMA combined with temozolomide on human glioblastoma cells growth in vivo experiments. Methods The nude mouse model with human glioblastoma cells subcutaneous transplantation was established. The mice were randomly divided into 4 groups to receive subcutaneous injection at the 14th day separately with: Normal saline 100 μl (control, n=8), Ad-PUMA 2×108 pfu/100 μl (PUMA group, n=8), 10 mg/kg TMZ (TMZ group, n=8) and 2×108 pfu/100 μl Ad-PUMA + 10 mg/kg TMZ (combined group, n=8). Mice were killed after 20 days treatment.Tumor volume, inhibition rates and apoptotic index (AI) were measured, meanwhile, apoptotic tumor cells were detected by TUNEL technology respectively. The expression of MGMT mRNA and MGMT protein were revealed by the methods of RT-PCR and Western blot. Results According to the order: control group, AdPUMA group, TMZ group, combined group, tumor volumes were (3.68±0.09), (2.63±0.13), (2.13±0.07),(0.97±0.02) cm3 respectively (P<0.05); the inhibitive rates were 0, 28.5 %, 42.1%, 73.6 % respectively and AI were (2.0±1.2) %, (11.4±2.6) %, (7.6±3.2) %, (20.6±8.6) % (P<0.05). The results of Western blot and RT-PCR showed that MGMT mRNA and MGMT protein levels in TMZ group were higher than other groups (all P<0.01). Conclusion Ad-PUMA combined with TMZ greatly enhances the sensitivity of human glioblastoma cells to TMZ and could effectively inhibit the proliferation and promote the apeptosis of glioblastoma cells, its mechanism was probably related Ad-PUMA promote apoptosis and inhibit MGMT expression.

16.
Chinese Journal of Neurology ; (12): 101-104, 2011.
Article in Chinese | WPRIM | ID: wpr-381926

ABSTRACT

Objective To explore the expression of vascular endothelial growth factor (VEGF)mRNA and protein in endothelia progenitor cells (EPCs) and VEGF in the culture medium and serum of patients during acute period of cerebral hemorrhage associated with hypertension (APCHH). Methods Mononuclear cells from peripheral blood of patients with APCHH ( 16 patients) and hypertension ( 16 patients) were isolated and induced to EPCs. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed to assay VEGF mRNA. VEGF protein was assessed by Western blotting. The VEGF protein level in patient's serum and culture medium ( at day 7 ) were assayed using VEGF ELISA Kit and compared between APCHH group and hypertension group. Results Compared with hypertension group, VEGF mRNA (0. 186 ±0. 035 versus 0.090 ±0.031, t =8.318, P <0.0l ) and protein (0. 223 ± 0. 028 versus 0.169 ± 0. 022, t = 3. 744, P < 0. 01 ) expression of EPCs, the concentration of VEGF protein in the supernatant (414 ±37 versus 316 ±29, t =8. 270, P <0. 01 ) and in serum (408 ±49versus 222 ±34, t = 12.406, P <0. 01 ) were all significantly increased in APCHH group. Conclusion The VEGF protein levels in serum of patients and in the culture medium, VEGF mRNA and protein expression in EPCs were all significantly increased during acute periods of cerebral hemorrhage.

17.
Article in Chinese | WPRIM | ID: wpr-385261

ABSTRACT

Objective To explore the influence of cerebral hemorrhage stress on endothelial progenitor cells(EPCs) proliferation and senescence. Methods Mononuclear cells from peripheral blood of patients with cerebral hemorrhage (CH group) and patients with the same underling diseases (control group) as the front were isolated and induced to EPCs. Proliferation, numbers of colonies and cells, senescence of EPCs were checked and compared between the two groups. Results Compared with control group, proliferation (0.270 ± 0. 074 vs 0. 185 ±0.050, t=3.802) ,numbers of colonies( (5. l0 ±0.79) cell/HPF vs ( 1.90 ±0.41 ) cell/HPF, t= 14.407) and cells( (75.15 ± 10.48) cell/HPF vs(57.10 ± 9.28 ) cell/HPF, t = 5. 157 ) of EPCs in CH group on the stress of cerebral hemorrhage were increased significantly (P < 0.01 ), but senescence of EPCs ( (45.54 ± 5.74 ) cells/100 cells vs ( 46.86 ± 5.06 ) cells/100 cells, t = 0. 686 ) in two groups were not different (P > 0.05 ). Conclusion Cerebral hemorrhage stress can increase proliferation of EPCs,but can not change the senescence destiny of EPCs.

18.
Journal of Chinese Physician ; (12): 735-738, 2008.
Article in Chinese | WPRIM | ID: wpr-399871

ABSTRACT

Objective To study that Geranylgeranylacetone(GA) induce the expression of heat shock protein 70 expression in hippocampus of Alzheimer's model rats and its effect on learning and memory ability in the model rats induced by Aβ1-42 injection into hippocampus.Method 72 health SD rats were randomly divided into GGA group,model group was injected with Aβ1-42 and control group was injected with normal saline into hippocampus.Y maze test was used to detect the learning and memory ability of the rats at the 7th,14th and 21st day after injection into hippocampus separately.HSP70 expression in hippocampus tissues were detected by RT-PCR and western-blot as soon as Y maze test has been finished.Result The learning and memory ability of the rats in model group decreased significantly at the 14th and 21st day after injection than those in control group(P<0.05),but not too much changed in GGA group(P<0.05).HSP70 expres- sions in hippocampus tissues in model group decreased gradually after injection Aβ1-42,but increased in GGA group at the 7th,14th and 21st day after injection.Conclusion GGA can induce the expression of HSP70 in hippocarnpus of Alzheimer's model rats and meliorate the neuron impairment as well as the learning and memory ability of the rats.

19.
Article in Chinese | WPRIM | ID: wpr-247294

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the changes in the expressions of inducible cyclooxygenase type 2 (COX-2) and membrane associated prostaglandin E-1(mPGES-1) in human carotid atherosclerotic plaques and to explore possible mechanisms of inflammatory process involved in plaque stability.</p><p><b>METHODS</b>The mRNA and protein levels of COX-2 and mPGES-1 were compared between minimally and grossly atherosclerotic arterial tissues. COX-2 and mPGES-1 gene expression were established by immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) in 10 mesenchymal artery controls and 24 atherosclerotic specimens. Presence of COX-2 and mPGES-1 protein was assessed by Western blotting.</p><p><b>RESULTS</b>Immunohistochemical staining showed that the COX-2 and mPGES-1 immunoreactive substances were present in the cytoplasm of smooth muscle cell. Compared with the control group, immunostaining positive cells increased in carotid atherosclerotic plaque group. COX-2 and mPGES-1 gene expression was significantly elevated in atherosclerotic plaques (P< 0.05, respectively). The increased mRNA and protein levels of COX-2 and mPGES-1 were correlated in atherosclerotic tissue (P< 0.05). The mRNA and protein levels of COX-2 and mPGES-1 related to degree of pathological damage in atherosclerotic tissue (P< 0.05). COX-2 and mPGES-1 were not found in the control group (mesenteric vascular walls).</p><p><b>CONCLUSION</b>COX-2 and mPGES-1 expression in plaques is significantly higher than that in the control group. These findings suggests that COX-2 and mPGES-1 might play a role in pathogenesis of atheroscleros and modulation of inflammatory process involved in plaque stability, and COX-2 may have proinflammatory enzyme properties.</p>


Subject(s)
Aged , Atherosclerosis , Genetics , Metabolism , Blotting, Western , Carotid Artery Diseases , Genetics , Metabolism , Cyclooxygenase 2 , Genetics , Metabolism , Female , Gene Expression , Humans , Immunohistochemistry , Intramolecular Oxidoreductases , Genetics , Metabolism , Male , Middle Aged , Prostaglandin-E Synthases , Reverse Transcriptase Polymerase Chain Reaction
20.
Article in Chinese | WPRIM | ID: wpr-676284

ABSTRACT

Objective To investigate the changes of the expressions of ATP-binding cassette transporter A1(ABCA1)and the retinoid X receptor(RXR?in human carotid atherosclerotic plaques and to explore the possible mechanisms by which ABCA1 affects the formation of carotid atherosclerosis(CAS). Methods 24 carotid atherosclerotic plaque and 10 intestinal artery specimens were respectively collected to compared the expression levels of ABCA1 mRNA.RXR?mRNA and those protein,ABCA1 and RXR?gene expressions were determined by reverse transcriptase polymerase chain reaction(RT-PCR)in the specimens,meanwhile the presence of ABCAI and RXRcprotein was assessed by Western blot.Results ABCA1(0.79?0.04)and RXR?(0.73?0.04)gene expression were significantly elevated in carotid atherosclerotic plaques(P

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