Artemin expression in chondrosarcoma and its effect on proliferation and migration of endothelial cells / 中国肿瘤生物治疗杂志

@#Objective: To investigate the expression of Artemin in chondrosarcoma and its effect on proliferation and migration of endothelial cells, and to explore the mechanism. Methods: A total of 40 chondrosarcoma tissue samples (low degree (Ⅰ), 20 cases; high degree (Ⅱ,Ⅲ), 20 cases) surgically resected from patients, who were treated in Lianyungang HospitalAffiliated to Nanjing University of Chinese Medicine from May, 2015 to April, 2019, were collected for this study. Another 20 cases of normal cartilage tissue specimen from patients with amputations due to car accidents were served as control. The expressions of Artemin, vascular endothelial growth factor (VEGF), Ki-67 and CD31+ vascular density in tumor tissues were detected by immunohistochemistry.After being treated with 10 ng/ml Artemin, the changes of VEGF, stromal cell derived factor-1 (SDF-1), matric metalloproteinase 2 (MMP2) and MMP9 in the supernatant of SW1353 cell culture were detected by enzyme-linked immunosorbent assay (ELISA), and the effects of Artemin-treated chondrosarcoma cells on the migration and proliferation of ECV304 cells were detected by Transwell migration assay and MTT cell proliferation assay, respectively. Results: The expressions of Artemin and Ki-67 in the tissues of low-level group were significantly higher than those in the control group (all P<0.01); the expressions ofArtemin and Ki-67 in the tissues of high-level group were significantly higher than those in the low-level group (all P<0.01). The expression of Artemin was positively correlated with VEGF level and vascular density in chondrosarcoma tissues (all P<0.01);Artemin promoted the secretion of VEGF by chondrosarcoma cells, but had no significant effect on the secretion of SDF-1, MMP2 and MMP9. Artemin induced the proliferation and migration of ECV304 cells by promoting the secretion of VEGF by chondrosarcoma cells (all P<0.01). Conclusion: Artemin is highly expressed in chondrosarcoma tissues and has a positive correlation with the expression of VEGF and vascular density. Artemin can enhance the angiogenesis induced by chondrosarcoma.

Stages Of Smoking Cessation Among Adult Current Smokers In Malaysia: Findings From The National E-Cigarette Survey 2016

@#Identification of the stages of smoking cessation among smokers is essential to improve the rate of smoking cessation. The aims of this study were to determine the prevalence and factors associated with stages of smoking cessation across the demographic distribution of adult smokers in Malaysia. Data were derived from a population-based study among Malaysian adults aged 18 years and above. Face-to-face interviews were carried out by trained staff using a validated questionnaire to obtain data related to smoking from 4,288 selected respondents. Of 4,288, 438 respondents are current smokers. Multivariable logistic regression analysis was used to determine factors associated with stage of smoking cessation. Approximately 60% (n=269/438) of the current smokers were in the pre-contemplation stage and 40% (n=169/438) were in the contemplation and preparation stages of smoking cessation. The proportion of pre-contemplators was higher among smokers with higher levels of nicotine addiction (71.3%), lower education attainment (71.4%), and those who were single/widowed/divorced (66.9%). Multivariable analysis showed that males, and those who reported low to high level of nicotine addiction were more likely to be in the pre-contemplation stage whilst those in the older age groups were more likely to be in the advanced stage of smoking cessation. The study revealed that the majority of current smokers in Malaysia had no intention to quit smoking within 6 months. Specific interventions targeting males, young adults and smokers with low to high nicotine addiction should be introduced to ensure the smokers proceed to the advanced stage of smoking cessation.

Mecanismo sensor y de adaptación a los niveles de oxígeno y su implicancia en las enfermedades cardiovasculares: a propósito del Premio Nobel de Fisiología-Medicina 2019 / Role of hypoxia-inducible factor-1 (HIF-1) in the adaptacion to oxygen levels: Nobel Prize in physiology-medicine 2019

RESUMEN: El Premio Nobel 2019 en Fisiología-Medicina se confirió a los Profesores Gregg Semenza, William Kaelin y Sir Peter Ratcliffe por sus investigaciones en la maquinaria molecular que regula la expresión de genes sensibles a los cambios en los niveles de oxígeno. La síntesis de eritropoyetina inducida por la disminución de los niveles sanguíneos de oxígeno condujo al estudio del gen de la eritropoyetina y descubrimiento de los elementos de respuesta a hipoxia (HRE) en la región promotora y posteriormente al factor transcripcional inducible por hipoxia tipo 1 (HIF-1). Este factor consta de dos subunidades: HIF-1α, sensible al oxígeno, y HIF-1β, expresada constitutivamente. HIF1 activa la transcripción de genes que codifican enzimas, transportadores y proteínas mitocondriales que disminuyen la utilización de oxígeno al cambiar el metabolismo oxidativo al metabolismo glicolítico y además aquellos involucrados en la angiogénesis y diferenciación celular. Las investigaciones paralelas en la enfermedad von Hippel-Lindau (VHL), un desorden autosómico dominante, permitieron descubrir el mecanismo de degradación de HIF1 en condiciones de normoxia y como se estabiliza bajo hipoxia. El impacto de HIF en clínica radica en el establecimiento de nuevas dianas terapéuticas para combatir la anemia y diversas enfermedades cardiovasculares. HIF promueve la angiogénesis a través de la expresión del factor de crecimiento vascular endotelial (VEGF), agente cardioprotector con potencial para tratar la isquemia/reperfusión, hipertrofia patológica e insuficiencia cardíaca.

ABSTRACT: The Nobel Prize in Physiology-Medicine was awarded to Drs. Gregg Semenza, William Kaelin and Sir Peter Ratcliffe for their research in the molecular machinery that regulates the expression of genes sensitive to the change in oxygen levels. The synthesis of erythropoietin induced by the decrease levels of oxygen in the blood led to investigate the promoter of the erythropoietin gene where the so-called hypoxia response elements (HRE) were described. Semenza et al. described a protein that binds to HREs and called it hypoxia-inducible transcriptional factor (HIF) that regulates gene expression among those involved in angiogenesis, cell differentiation and glycolytic enzymes. HIF presents two oxygen-sensitive subunits HIF-1α and HIF-1β constitutively expressed. In parallel, Kaelin et al. investigated von Hippel-Lindau disease (VHL), an autosomal dominant disorder, discovering a mutation of this protein generated a behavior similar to hypoxia. The impact of HIF-1α lies in the search for new strategies such as hydrolase inhibitors to combat prevalent diseases, including anemia and cardiovascular diseases These compounds promote the expression of vascular endothelial growth factor (VEGF), a cardioprotective agent with potential use in pre- and post-conditioning therapy, cardiac hypertrophy and heart failure.

Breech presentation at term: outcomes and mode of delivery in a tertiary care teaching hospital In South India

The aim of this study was to study the outcomes of all patients who presented with breech presentation at term (≥37 weeks), to assess what percentage of patients were offered External cephalic version (ECV), the rates of success of the procedure and the rates of vaginal delivery following successful ECV. It was a retrospective study of 669 patients diagnosed with breech at term, their clinical records were retrieved and data like age, BMI, parity, type of breech and scan findings noted. ECV was done in 256 patients and was successful in 35.5% of women with 51.1% being multigravidas and 26.8% in primigravidas. 76.9% of women with successful ECV delivered vaginally.  There was no significant fetal or maternal morbidity documented as a result of ECV in this study.

Clinical Characteristics and Outcomes of 80 Pregnant Women after External Cephalic Version / 中山大学学报(医学科学版)

[Objective] To investigate the safety and associated factor of external cephalic version (ECV) in third trimester,and to enrich clinical experience to improve the successful rate and lower cesarean section (CS) rate.[Methods] 80 pregnant women conducting ECV in third trimester in the third affiliated hospital of Sun Yat-sen University from September 2015 to July 2017 were enrolled in our study.Divided to successful group and failing group,we compared the clinical characters and pregnancy outcomes.[Results] Of the 80 pregnancy,48 women (60.0％) succeed with cephalic presentation.Compared to the failing group,the successful group is statistically different in parity,BMI and amniotic fluid depth.In the failing group,all women underwent CS with 3/48 in successful group.No women conducted ECV complicated fetal distress and emergency CS,premature rupture of membranes complicated in 11 (13.8％) cases in all women.[Conclusions] ECV is safe for mother and fetus.Encouraging the suitable pregnancy women to conduct ECV and enhancing clinical skills can improve ECV success rate.

Construction and assessment of recombinant plasmid pRNAT-U6.1/CFB siRNA and its inhibitory effect on proliferation of human umbilical vein endothelial cells / 中华实验眼科杂志

Background Choroidal neovascularization (CNV) is one of the causes of blindness in multiple eye diseases.Researches showed that complement system participates in the pathogenesis of CNV.Objective This study was to construct the recombinant of complement factor B-small interference RNA (CFB-siRNA) expression vector and to observe its inhibitory effect on human umbilical vein endothelial cells (ECV-304).Methods CFB gene primers were designed based on human CFB gene,and an expression vector of CFB-siRNA was constructed by inserting CFB-siRNA into pRNAT-U6.1/Neo plasmid.Recombinant plasmids were confirmed by the digestion analysis of restriction endonuclease,and all inserted sequences were verified by DNA sequencing.The recombinant pRNAT-U6.1/CFB-siRNA plasmid and the blank plasmid were transfected into ECV-304 cells in the CFB-siRNA group and blank plasmid group by electroblot,respectively,and non-transfected cells served as the normal control group.The cells were observed under the fluorescence microscope 48 hours after transfection,and the transfective efficiency was calculated.The relative expression of CFB mRNA in the cells of different groups was detected by semi-quantitative reverse transcription PCR (RT-PCR).MTT was employed to calculated the growth inhibitory rates of the cells 24,48 and 72 hours after transfection.The percentages of the cells in different cell cycles were detected by flow cytometry.Results The sequence of the target vector was identical to the designed sequence.The green fluorescence protein (GFP) was seen in both the CFB-siRNA group and the blank plasmid group.The relative expression levels of CFB mRNA were 0.07 ±0.04,0.14 ±0.02 and 0.14 ±0.03 in the CFB-siRNA group,the blank plasmid group and the normal control group,respectively,a significant difference was obtained among the three groups (F=233.05,P =0.00);the expression level of CFB mRNA in the CFB-siRNA group was significantly declined in comparison with the blank plasmid group and the normal control group (both at P＜0.05).The growth inhibitory rates of the cells were (23.45 ±0.01) ％,(33.48 ±0.02) ％ and (45.49±0.01) ％ at 24,48 and 72 hours after transfection,respectively,a significant difference was obtained among the three groups (Fgroup =212.99,P =0.00);the growth inhibitory rates in CFB-siRNA group were significantly higher than that in the blank plasmid group and normal control group (all at P＜ 0.05).The percentages of G1 phase cells were (44.4 ±0.5) ％,(25.8 ±0.4) ％ and (27.9 ± 0.6) ％ in the CFB-siRNA group,the blank plasmid group and the normal control group respectively,a significant difference was obtained among the three groups (F=58.98,P=0.00).The percentages of G1 phase and G2 phase cells in the CFB-siRNA group were significantly higher than those in the blank plasmid group and the normal control group (all at P＜0.05).Conclusions Recombinant pRNAT-U6.1/CFB siRNA inhibits the proliferation of ECV-304 cells effectively by arresting the cells in G1 intermediate phase of the growth cycle.

Polimorfismo de los genes C677T de la enzima metil-n-tetrahidrofolato reductasa, A2756G de la enzima metionina sintasa y A66G de la enzima metionina sintasa reductasa y su relación con la homocisteína en pacientes con enfermedades cardiovasculares / C677T gene polymorphism of the enzyme-n-methyl-tetrahydrofolate reductase, A2756G gene polymorphism of the enzyme methionine synthase and A66G gene polymorphism of methionine synthase reductase and its relationship with homocysteine in patients with cardiovascular disease

La Hiperhomociteinemia (HHcy) se considera como un factor de riesgo idenpendiente para el desarrollo de aterosclerosis y de enfermedad arterial coronaria (EAC). Los polimorfismos en encimas involucradas en la regulación del metabolismo de la Hcy como la metiltetrahidrofolato reductasa (MTHFR), metionina sintasa (MTR), y la metionina sintasa reductasa (MTRR) pueden contribuir a la variación de los niveles de homocisteína en plasma (Hcy). En este estudio investigamos la asociación de los polimorfismos genéticos de las enzimas involucradas en la remetilación de la homocisteínia: metionina sintasa (MTR), metil N-tetrahidrofolato reductasa (MTHFR) y metionina sintasa reductasa (MTRR), con los niveles de Hcy en pacientes con EAC y sujetos sanos. Población 136 individuos de los cuales 90 presentaron diagnóstico de enfermedad cardiovascular (IAM y ACV) y 46 eran aparentemente sanos (controles). La concentración de Hcy fue significativamente más alta en pacientes con ECV que en los sujetos control (P<0,001). HHcy (>15 µmol/L) confirió un RR de IAM de 2.52 (95% IC: 1.4-4.4, P<0,001) y de ACV de 1.88 (95% IC: 1.0-3-5, P<0,05). Los niveles de vitamina B12 y folato se encontraba en el rango de los valores de referencia en el 86% de los individuos. La frecuencia del alelo T para C677T de MTHFR, del alelo G para A66G de MTRR y del alelo G para A2756G de MTR fueron 0.31, 0.30, 0.22 respectivamente para los sujetos casos. Los polimorfismos C677T, A66G y A276G de los genes MTHFR, MTRR y MTR no tuvieron diferencia estadísticamente significativa entre el grupo de casos con respecto al grupo control. Los polimorfismos estudiados no se relacionaron estadísticamente con la HHcy en los individuos en estudio. Sugerimos que HHcy confiere riesgo para ECV. En nuestro estudio encontramos evidencia de que la regulación de Hcy es poligénica.

The Hiperhomocysteinemia (HHcy) is considered an independent risk factor for developing atherosclerosis and cardiovascular arterial disease (CAD). The polymorphisms of the enzymes involved in the regulation of homocysteine (Hcy) metabolism as the methyltetrahydrofolate reductase (MTHFR), methionine synthase (MTR) and methionine synthase reductase (MTRR) may contribute to the elevation of Hcy in plasma. The main aim of the this study was to investigate the association of genetic polymorphisms of the enzymes involved in remethylation of homocysteine: methionine synthase (MTR), N-methyl tetrahydrofolate reductase (MTHFR) and methionine synthase reductase (MTRR), with levels of Hcy in patients with CHF and healthy subjects. Population: 136 subjects of whom 90 had a diagnosis of cardiovascular disease (heart failure and stroke) and 46 were apparently healthy (controls). The concentration of Hcy was significantly higher in patients with cardiovascular disease than compare with the control group (P<0.001). HHcy (>15 mmol/L) conferred a relative risk (RR) of heart failure of 2.52 (95% CI: 1.4-4.4, P <0.001) and stroke, RR of 1.88 (95% CI: 1.0-3.5, P <0.05). The levels of vitamin B12 and folate was in the range of reference values in 86% of subjects. The frequency of the T allele of MTHFR C677T was 0,31, for the G allele of MTRR A66G was 0,30 and for the G allelwe for MTR A2756G was 0.22 for the subjects with heart failure and stroke referred as case. C677T polymorphism, A66G and A2756G of the genes MTHFR, MTRR and MTR had no associated statistically with HHcy in the subjects of the study. We suggest that HHcy confers risk for CVD. In our study we found evidence that the regulation of Hcy is polygenic.

Patrón de comportamiento tipo A, ira y enfermedades cardiovasculares (ECV) en población urbana chilena / Type A Behavior Pattern, Anger and Cardiovascular Disease (CVD) in Chilean Urban Population

Un aumento de las enfermedades cardiovasculares (ECV) mundialmente, la existencia de un porcentaje más alto que el promedio nacional de éstas para la región de Maule (Chile), y una atención incrementada en el análisis de factores psicológicos, motivan el análisis del Patrón de Comportamiento Tipo A (PCTA) y la ira respecto de las ECV. Se trabajó con 1007 participantes de 18 a 74 años (ciudadanos de Talca, Chile), mayoritariamente mujeres, quienes respondieron un cuestionario (información socio demográfica, hábitos alimentarios y de estilo de vida), la Escala Retiro de Patrón de Conducta tipo A (ERCTAa), y el Inventario de Ira de Novaco. Se les midió peso, masa corporal, presión arterial y sangre, como factores de riesgo cardiovascular. Los participantes son altamente sedentarios (79.9%), tabáquicos (53.6%), hipercolesterolémicos (44.5%), con sobrepeso (40.7%) y obesidad (32.6%), un cuarto de los cuales presenta hiperglicemia e hipertensión y con PCTA equirepartido según sexo. Es la ausencia de PCTA (ó presencia de PCTB) la que aparece asociada a factores tradicionales de riesgo cardiovascular (FRCV). La ira alta se presenta más en mujeres que en hombres (2.1% vs. 0.3%; c²(3) = 27.99, p<.0001), disminuyendo para ambos sexos con la edad, pero los infartos acaecen igualmente según sexo.

A worldwide raise in the number of cardiovascular disease (CVD) and the existence of a higher percentage in Maule (Chile) than the national media, and increased attention in the analysis of psychological factors motivate to analyze the Type A Behavior Pattern (TABP) and anger in relation to CVD. The sample was 1007 adults between 18 and 74 years old (citizens of Talca, Chile), mostly women. They provided information about their demographic details, eating habits and lifestyle, answered the Novaco's Anger Inventory and the Retiro Scale of Type A Behavior (RSTAB), and also were taken measurements like weight, body mass index and blood pressure and blood tests related to risk factors to traditional cardiovascular diseases. The results show Overall, that the participants appear highly sedentary (79.9%) with relatively high levels of tabaquism (53.6%), and hypercholesterolemia (44.5%), overweight (40.7%) and obesity (32.6%). A quarter of the sample also presents hyperglycemic indexes, hypertension and TABP unequally distributed by sex. The absence of PCTA (or PCTB presence) appeared mostly associated with traditional cardiovascular risk factors (CRF). Regarding anger, women present more high than men (2.1% against 0.3%; c²(3) = 27.99, p<.0001), decreasing for both sexes with age, while also befall stroke by sex.

Effects of methotrexate enantiomers on ECV304 cell inhibition and its mechanisms / 中国药理学通报

Aim To investigate the effect of MTX(included(±)MTX,(+)MTX and(-)MTX)on the proliferation of ECV304 cells and to explore its mechanisms.Methods ECV304 cells were cultured.The cell proliferation was determined by MTT.The morphological changes were inspected by inverted microscope.Cell cycle phases were assayed by propidium iodide staining flow cytometry.Results ECV304 cells were treated with(+)MTX,(-)MTX and(±)MTX at 1～150 μmol·L~(-1) for 24,48,72 h.The results showed that the proliferation of ECV304 cells was significantly inhibited under different conditions.The order of the inhibited efficacy was(+)MTX>(±)MTX>(-)MTX.The morphology of ECV304 cells were changed by(+)MTX,(-)MTX and(±)MTX treatment,which included the cell shrinkage,chromatin condensation.After administration of 10 μmol·L~(-1) of(+)MTX,(-)MTX and(±)MTX for 48 h,the cell cycle phases were assayed by propidium iodide staining flow cytometry.The result showed DNA replication was interfered by(+)MTX,(-)MTX and(±)MTX treatment.Conclusions The proliferation of ECV304 cells has the chiral selective effects by(+)MTX and(-)MTX treatment,and the inhibition on ECV304 cells proliferation of(+)MTX is significantly stronger than that of (-)MTX.

Pseudoxantoma elástico con complicaciones tardías y asociaciones infrecuentes / Pseudoxanthoma elasticum with late complications and unusual associations

El pseudoxantoma elástico (PXE) es un trastorno hereditario poco común del tejido conectivo, que se caracteriza por lesiones cutáneas, oculares y cardiológicas originadas por fragmentación y calcificación de las fibras elásticas. La mayoría de casos se hereda con un patrón autosómico recesivo, y, en menos proporción, autosómico dominante. El propósito del presente reporte es dar a conocer un caso de pseudoxantoma elástico con complicaciones tardías y asociaciones infrecuentes, como son una ECV (enfermedad cerebro vascular) recidivante, abortos recurrentes, la presencia de un bocio multinodular unilateral derecho, fibromatosis uterina gigante no sintomática, hipertensión arterial, y un antecedente no bien clarificado con respecto a la presencia de diabetes mellitus.

Pseudoxanthoma elasticum (PXE) is connective tissue's rare inherited disorder, characterized by skin, eye and cardiac lesions, originated by fragmentation and calcification of elastic fibers. Most cases are inherited with an autosomal recessive pattern, and in a little proportion, with an autosomal dominant pattern. This paper has the purpose of reporting a case of pseudoxanthoma elasticum with late complications and unusual associations, such as recidivated stroke, recurrent aborts, right multinodular unilateral goiter, giant uterine fibromatosis, arterial hypertension, and an unclear antecedent of diabetes mellitus.

Effect of β-asarone on expression of ECV340 cell adhesion molecules induced by β-amyloid peptide / 中成药

AIM:To explore the effect of β-asarone on vascular endotheliam and adhesion molecule expression of endothelium induced by β-amyloid peptide from Alzheimer's disease and to estimate the injury repair.METHODS:Cultured ECV304 cells were incubated with freshly solublizeal Aβ1-42 and the mixture of Aβ1-42 and β-asarone,the expression of three central adhesion molecules,CD106,CD62P,CE62E and Ca2+ concentration were examined and apoptosis was recorded by Flow eytometry.Test viability of cells by MTT methods.RESULTS:The results showed that in model group and treated group,ligation of endothelial CD106,CD62P,CE62E,markers for endothelial cell activation and Ca2+ concentration,leads to a lot of release.The livability decreased and the apoptosis increased.Further more,simultaneous treatment of ECV304 cells with β-asarone resulted in the decrease significandy in these three adhesion molecules described above and Ca2+ concentration as well as the livability upper and apoptosis lower.CONCLUSION:CD106,CD62P,CE62E,important inflammational factor of Aβ-induced endothelial injury,may be promotion of the inflammatory scade in vascular endothelial.β-asarone may protect ECV304 cell apoptosis by regulate Ca2+ and expression of cell surface markers.

Influence of RNA interference-induced silencing Smad4 gene on proliferation and migration of ECV304 cell line / 肿瘤

Objective: To investigate the effect of silencing Smad4 gene expression by using small interfering RNA (siRNA) on proliferation and migration of ECV304 cell line and explore the role of Smad4 in angiogenesis. Methods: Two siRNA sequences targeting Smad4 gene including siRNA1 and siRNA2 were designed and then synthetized. The two sequences of siRNA were transfected into ECV304 cells via lipofectamine mediation. The alteration of Smad4 mRNA and protein expression was examined by real-time RT-PCR and Western blot, respectively. The proliferation capacity of ECV304 cells was measured by cell cycle analysis and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) flow cytometry. The monostratal wound healing test was used to determine the migration capacity of ECV304 cells. Results: siRNA2 was successfully screened out of the two sequences. Transfection of siRNA2 significantly knocked down the Smad4 mRNA and protein expressions. The proliferation and migration capacity of ECV304 cells are significantly enhanced after transfection with siRNA2. Conclusions: Highly effective and specific siRNA targeting Smad4 gene can be successfully screened by RNA interference technique. Selective silencing of Smad 4 gene greatly promotes the proliferation and migration of ECV304 cells.

Study on VVC recombinant protein cytolytic activity mechanism of an extracellular cytolytic toxin of Vibrio vulnificus / 中华微生物学和免疫学杂志

Objective To investigate the cytolytic activity of extracellular cytolytic toxin rVVC of Vibrio vulnificus on the apoptosis of human ECV304 cells, and to analyze the activities of Caspase-3,-8 and -9. Methods The cytotoxic effect of refolded rVVC on the growth and apoptosis of ECV304 cells was identified by MTT, Hochest33342/PI fluorescent staining, flow cytometry and DNA agarose electrophoresis analysis, respectively. The activities of Caspase-3, -8 and -9 was measured using a colorimetric method. Results The viability of human ECV304 cells exposed to rVVC was inhibited by rVVC after 24 h. 2.0 HU/ml rVVC groups had a better cytotoxic effect to human ECV304 than that of 0.5 HU /ml rVVC groups. The apoptosis of human ECV304 cells in 2.0 HU/ml rVVC+40 μmol/L Z-VAD-FMK groups was relative reduced than that of 2.0 HU/ml of rVVC groups. After 0.5 h treatment with 2.0 HU/ml of rVVC, the Caspase-3 activity in human ECV304 cells increased gradually and reached the peak at 3 h (versus control groups, P<0.01). The activity of Caspase-8 and -9 remained unchanging. Conclusion The rVVC has cytotoxic effect on human ECV304 and the cytolysin is probably correlated with Caspase-3.

Effect of Docosahexaenoic Acid (DHA) on the Apoptosis of Human Endothelial ECV304 Cells

DHA, one of w-3 fatty acids, modulates cell growth or death though the changes of apoptotic signaling in human endothelial ECV304 cells. We investigated the effects of DHA on the changes of apoptotic signaling in human vascular endothelial ECV304 cells using lipid peroxidation (LPO) metabolites. LPO could be originated by dietary polyunsaturated fatty acids such as linoleic acid (LA), arachidonic acid (AA) and docosahexaenoic acid (DHA). DHA caused cell death of ECV304 cells compared to LA, AA or control as evidenced by changes in cell morphology and MTT assay. LPO levels was significantly elevated by 10 fold in DHA-treated ECV 304 cells and caspase-3 activity was increased by DHA corresponding to increasing incubation times compared to control. One of reasons of the cell death in DHA-treated ECV304 cells could be expected that caspase activity, marker for mitochondrial damages, might be triggered by the increasing LPO levels. Our results strongly indicated that DHA induced LPO production has an important role on apoptotic signaling pathway in ECV304 cells. LPO production in endothelial cells which was metabolized by oxidation of dietary PUFA, might be one of risk factors in the initial progression of atherosclerosis.

Effects of Ultra-filtration Extract from Angelica Sinensis and Hedysarum Polybotrys’s Mixture on VEGF mRNA Expression of ECV-304 Cell / 中国中医药信息杂志

Objective To study the effect of the ultra-filtration extract from Angelica sinensis and Hedysarum polybotrys’s mixture on the VEGF mRNA expression of ECV-304 cell. Method Angelica sinensis and Hedysarum polybotrys’s mixture were refined by ultra-filtration technique. ECV-304 cells were cultured as model, and its proliferation was detected by MTT colorimetry. VEGF mRNA expression was observed by semi-quantitative RT-PCR. Results The ultra-filtration extract from Angelica sinensis and Hedysarum polybotrys’s mixture could markly promote the growth of ECV-304 cells, there was significant difference between experimental and control group (P

Screening the Effective Component of Rhizoma Tupistrae Chinensis with Inhibiting Angiogenesis / 中国中医药信息杂志

Objective To discover the effective component of Rhizoma Tupistrae chinensis with inhibiting angiogenesis. Methods Several parts of Rhizoma Tupistrae chinensis were obtained by using different extraction solvents. They were screened by the model of ECV304 cell membrane chromatography with anti-VEGF antibody as a control drug. The inhibition of effective component (ZGQ-C) on ECV304 proliferation in vitro was examined by MTT assay. Results The inhibition rate (40.51%,51.11%,63.54%,74.32%,81.26%) was correlated with the ZGQ-C concentration. The ECV304 cell had significantly changed in morphology. Conclusion The ZGQ-C is an effective component for inhibiting angiogenesis. The screening results on the model have a good correlation with that of MTT assay.

Role of Oxygen Free Racical in the Proliferation of ECV304 Induced by AngⅡ / 昆明医科大学学报

Objective To explore the role of oxygen free radicals in the proliferation of ECV304 induced by AngⅡ.Methods The lines of human umbilical vein endothelial cell(ECV304)cultured in vivo were divided into three groups which were treated by AngⅡ,AngⅡ+N-acetyl-L-cysteine(NAC),and normal culture medium.First we observed the proliferous effect of ECV304 induced by AngⅡat different concentration with improved MTT and microscope.Then the contents of oxygen free radicals(?OH)in three groups were detected by spectrophotometer.Results ECV304 incubated with AngⅡ(0.03125～1?mol/L)for 12 hours increased the proliferation rate(P 0.05 vs.control group).Conclusions ECV304 induced by AngⅡcan produce oxygen free radicals(?OH),and the contents of oxygen free radicals(?OH) increase with the prolongation of time and the enlargement of dose;Antioxidant NAC can inhibit the proliferation of ECV304 induced by AngⅡ,this effect may be related with reducing the content of oxygen free radicals(?OH);oxygen free radicals(?OH)may be one of the major mocleculars which play an important role in the signal transduction of ECV304 proliferation.

Effect of Recombinant N-segment of Thrombospondin-1 on the Proliferation of ECV304 / 中国生物工程杂志

Objective: To construct an expression system containing the N-terminal segment of Thrombospondin-1 (TSP-1) in E. coli. To investigate the activity of TSF. Methods: thbs1 gene fragment was amplified from human fetal cord vein endothelial cells and inserted into plasmid pET32c ( + ) which was then transfected and expressed in E. coli. Investigate the effect of recombinant TSF to the proliferation of ECV304 in vitro with MTT. The expression level of CD36 was assayed by FACS. Results: Recombinant TSF was purified. TSF could restrain the proliferation of ECV304 with CD36 low-expression. Conclusions:Low dose of rTSF was a latent assistant treatment of anti-cancer.

Entery of calcium into ECV304 endothelial cell via CaCa~(2+) activated non-selective cation channel / 中国药理学通报

AIM To investigate the pathways of Ca 2+ entry into ECV304 endothelial cell and the effect of angiotensin Ⅱ(AⅡ) on calcium activated non setective cation channel(CAN). METHODS The cell attachment and whole cell configurations of patch clamp technique were used to record channel activity. RESULTS (1) The single channel conductance is ? o=(12 90?2 11) pS( n =4) for Ca 2+ passing through CAN of ECV304 cell in condition of pipette solution without K + and Na + but composed 120 mmol?L -1 CaCl 2. The channel current amplitude and open time can be enhanced by 1?10 -7 mol?L -1 AⅡ. The enhanced conductance in CAN is ? 1=(22 18?2 29) pS( n =4). The results of whole cell recording are identified with single channel recording. (2) The whole cell configuration was carried out for recording voltage dependent Ca 2+ channel in ECV304 cell. The peak current amplitude was (29 32?3 56) pA( n =4). This current was inhibited to (6 00?3 94) pA( n =4) by nifedipine and activated by BayK8644. CONCLUSIONS (1)Ca 2+ enters ECV304 cell via Ca 2+ activated non selective cation channel and voltage dependent L type calcium channel. (2) AⅡ can significantly enhance the calcium entry via CAN in ECV304 cell.

The effect of Tanshinone IIA on the expression of NF-?B、I?B-? and the mRNA of ICAM-1 and VCAM-1 of ECV304 induced by TNF-? / 中国药理学通报

Aim To illustrate the role of Tanshinone IIA(TanIIA)extracted from dan-shen root in atherosclerosis and its different targets by investigating the effect on the expression of NF-?B、I?B-? and the mRNA of ICAM-1 and VCAM-1 in human umbilical vascular endothelial cell 304(ECV304).Mehods The TNF-?-induced cultured ECV-304 cell injury model was established to analyze the expression of NF-?B、I?B-? by cell ELISA method and the mRNA of ICAM-1 and VCAM-1 by RT-PCR method of the cells cultured with different denses of TanIIA.Results The results of ELISA method showed that the expression of the NF-?B was reduced significantly and the expression of the I?B-? was enhanced significantly in ECV304 which were cultured with higher denses of TanIIA.The results of RT-PCR method showed that the mRNA of ICAM-1 and VCAM-1 induced by TNF-? was significantly refrained.Conclusions TanIIA has a significant effect on inhibiting the mRNA of ICAM-1 and VCAM-1 induced by the NF-?B activation,so it can play an impotant role in resisting atherosclerosis.