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Objective@#To investigate whether platelet-derived growth factor-BB (PDGF-BB) can regulate phenotypic transformation of pulmonary artery smooth muscle cells (PASMCs) via SIRT3 affecting glycolytic pathway.@*Methods@#The PASMCs were isolated from Sprague Dawley rats. PASMCs were divided into 3 groups by using 2-deoxyglucose (2-DG), an inhibitor of the glycolytic pathway: normal control group, PDGF-BB group(30 ng/ml) and PDGF-BB (30 ng/ml)+2-DG (10 mmol/L) group. In lentivirus-mediated overexpression assay, cells were divided into control group, PDGF-BB group(30 ng/ml), PDGF-BB+deacetylase sirtuin-3 (SIRT3) overexpression group and PDGF-BB+empty vector group. The expression levels of phenotype related index such as α-smooth muscle actin (α-SMA), smooth muscle myosin heavy chain (SM-MHC), calponin, vimentin were detected by qRT-PCR and Western blot. Meanwhile, the expression of α-SMA was detected by cellular immunofluorescence staining. EDU staining was used to detect the proliferation of PASMCs. The expression of SIRT3 was detected by Western blot. The expressions of glucose transporter 1 and aerobic glycolytic enzymes were detected by qRT-PCR and Western blot in lentivirus-mediated overexpression assay.@*Results@#(1) PDGF-BB affects PASMCs phenotypic transformation through glycolytic pathway: compared with normal control group, PDGF-BB significantly decreased the expressions of contractile phenotype markers such as α-SMA, SM-MHC, calponin mRNA and protein (all P<0.05), but it increased the expressions of the synthetic phenotype marker vimentin mRNA and protein (both P<0.05). Cellular immunofluorescence assay showed that PDGF-BB significantly decreased the number of α-SMA positive cells, while 2-DG reversed the process. (2) PDGF-BB promoted cell proliferation through glycolytic pathway: the proliferation of PASMCs was significantly higher in PDGF-BB group than in control group (P<0.05), and which could be significantly reduced by 2-DG (P<0.05). (3) PDGF-BB inhibited the expression of SIRT3 protein in PASMCs: the expression of SIRT3 protein in PDGF-BB group was lower than that in control group (P<0.05). (4) PDGF-BB affected glycolytic pathway through SIRT3:compared with the control group, PDGF-BB significantly increased the expression levels of glucose transporter 1 (Glut1), hexokinase 2 (HK2) and 6-phosphfructo-2-kinase 3 (PFKFB3) mRNA (all P<0.05), which was reserved by over-expression of SIRT3. There were no significant difference in mRNA expression levels between PDGF-BB group and PDGF-BB+empty vector group (P>0.05).Compared with the control group, PDGF-BB significantly increased the expression levels of Glut1, HK2 and PFKFB3 protein(all P<0.05), which was reserved by over-expression of SIRT3. There were no significant differences in protein expression levels between PDGF-BB group and PDGF-BB+empty vector group (all P>0.05).@*Conclusion@#PDGF-BB regulates phenotypic transformation of PASMCs via SIRT3 affecting glycolytic pathway.
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Objective@#To investigate the feasibility and efficacy of the establishment of regional cooperative acute ST-segment elevation myocardial infarction (STEMI) rescue network among the prefectural-level city hospitals in China.@*Methods@#Based on real-time remote electrocardiogram transmission and "120" emergency systems, we established a regional collaborative STEMI treatment network with our hospital as the network unclears including 8 second-class affiliated hospitals of Jiangsu University in 2013. STEMI treatment time, therapeutic effects and economic indexes were compared before (from January 2010 to December 2012, 180 cases, pre-network) and after (From January 2013 to December 2015, 374 cases, post-network) the establishment of the regional collaborative STEMI treatment network.@*Results@#Post establishment of the rescue network, mean first medical contact (FMC) to balloon (FMC-to-B) time, referral time and obtaining informed consent time were all significantly decreased from (191±41), (94±18), (25±9) minutes to (93±19), (53±18), (7±5) minutes, respectively, in comparison with the pre-network era(all P<0.05). There was a trend of prolonged FMC-to-B time in proportion to aging of STEMI patients(trend P<0.05). Three months post discharge, LVEF was higher (55.3%±10.7% vs. 48.8%±12.1%, P<0.05) and LVEDd was lower ((49.1±10.8)mm vs.(51.8±9.2)mm, P<0.05) in the post-network group compared to pre-network group.In-hospital mortality was also significantly reduced post the establishment of the rescue network (2.14%(8/374) vs. 3.89%(7/180), P<0.05). The results also showed that the total costs (42 017(25 069, 75 148)yuan vs.51 030(28 137, 105 861)yuan), days of hospitalization ((9.1±4.5) days vs. (15.3±4.8)days) and percentage of medicine and consumables were all significantly decreased in the post-network group compared to pre-network group(all P<0.05).@*Conclusion@#Establishment of the regional cooperative rescue network is feasible among the prefectural-level city hospitals in China. Establishment of such network can improve the prognosis and decrease the FMC-to-B time, the rate of in-hospital mortality and financial burden of patients with STEMI, and serves as an effective strategy to improve the rescue ability for STEMI patients.
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Objective@#To explore the molecular mechanism of docosahexaenoic acid (DHA) on regulating the phenotype switching of hypoxia-induced pulmonary arterial smooth muscle cells (PASMCs).@*Methods@#The PASMCs were isolated from Sprague Dawley rats. PASMCs were divided into five groups: normal control group, hypoxia group (1%O2, 94%N2, 5% CO2 stimulation for 12 hours), hypoxia+ DHA group (10 μmol/L DHA pretreatment followed by 12 hours hypoxia), hypoxia+ DHA+ NFATc1 overexpression group (transfection of the NFATc1 lentivirus for 24 hours, followed by hypoxia stimulation for 12 hours after 10 μmol/L DHA treatment), and hypoxia+ DHA+ siNFATc1 group (transfection the siNFATc1 for 24 hours, followed by hypoxia stimulation for 12 hours after 10 μmol/L DHA treatment). The hypoxia stimulation was achieved by use of a special hypoxia incubator (1%O2, 94%N2, 5%CO2). The expressions of NFATc1 of various groups were determined by qRT-PCR and Western blot. The expression of α-SMA was determined by immunofluorescence staining, qRT-PCR and Western blot. The expression of SM22 was determined by qRT-PCR. The proliferation of PASMC was determined by EDU staining.@*Results@#The mRNA and protein expression levels of NFATc1 were significantly upregulated in hypoxia group compared with the normal control group (P<0.05), while hypoxia-induced upregulation of NFTAc1 could be significantly downregulated by DHA treatment (P<0.05). The α-SMA positive cell number, protein and mRNA levels of α-SMA and the mRNA level of SM22 were significantly lower in the hypoxia group than in normal control group, which could be significantly reversed by DHA, the protective effects could then be abolished by NFATc1 overexpression. Above indices were significantly lower in the hypoxia+ DHA+ siNFATc1 group than in hypoxia+ DHA+ NFATc1 overexpression group (P<0.05). The proliferation of PASMCs was significantly higher in the hypoxia group than in the control group (P<0.05), and which could be significantly reduced by DHA (P<0.05), and the protective effect of DHA could be significantly abolished by overexpression of NFATc1 (P<0.05). The proliferation of PASMCs was significantly lower in the hypoxia+ DHA+ siNFATc1 group than in the hypoxia+ DHA+ overexpression NFATc1 group (P<0.05).@*Conclusion@#DHA could prevent hypoxia-induced PASMCs phenotype switching and proliferation by inhibiting NFATc1 signaling.
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<p><b>OBJECTIVE</b>To observe whether CD137 signaling could affect the nuclear factor of activated T cells c1 (NFATc1) expression through nuclear factor-κB (NF-κB) pathway in mice aortic vascular smooth muscle cells (VSMCs).</p><p><b>METHODS</b>Adherence methods for tissues explants were used for primary culture of mouse aortic VSMCs. The mRNA expression of CD137 and NFATc1 was detected by real-time quantitative PCR (RT-qPCR). The VSMCs protein expression of IκB-α, NF-κB p65, phospo-p65 and NFATc1 was determined by Western blot. The level of CD137 was measured by Flow Cytometry (FCM).</p><p><b>RESULTS</b>(1) The mRNA and protein expression of CD137 in VSMCs was significantly upregulated at 24 h after co-culture with TNF-α (10 ng/ml, all P < 0.05). (2) Compared with the control group, the level of p-NF-κB p65 in cytoplasm and nucleus was significantly increased (8.34 ± 0.28 vs. 1, P < 0.05, and 2.64 ± 0.42 vs. 1, P < 0.05) while the level of IκB-α was reduced (1 vs. 2.70 ± 0.28, P < 0.05) after co-treatment with agonist-CD137 mAb, above effects were partly blocked by adding specific NF-κB inhibitor PDTC (30 µmol/L: 1.15 ± 0.14 vs. 8.34 ± 0.28, P < 0.05, and 2.09 ± 0.12 vs. 2.64 ± 0.42, P < 0.05, and 1.78 ± 0.74 vs. 1, P < 0.05). (3) The mRNA (2.07 ± 0.09 vs. 1, P < 0.05) and protein (1.75 ± 0.07 vs. 1, P < 0.05) expression of NFATc1 was significantly upregulated by agonist CD137mAb compared with the control group, and these effects could be reversed by PDTC (1.15 ± 0.07 vs. 2.07 ± 0.09, P < 0.05, and 0.90 ± 0.11 vs. 1.75 ± 0.07, P < 0.05).</p><p><b>CONCLUSION</b>CD137 signaling could affect the NFATc1expression in VSMCs through NF-kappaB pathway.</p>
Subject(s)
Animals , Mice , Cells, Cultured , I-kappa B Proteins , Muscle, Smooth, Vascular , Metabolism , Myocytes, Smooth Muscle , NF-KappaB Inhibitor alpha , NF-kappa B , Metabolism , NFATC Transcription Factors , Metabolism , RNA, Messenger , Signal Transduction , Tumor Necrosis Factor Receptor Superfamily, Member 9 , Metabolism , Tumor Necrosis Factor-alpha , Up-RegulationABSTRACT
Objective To evaluated the effect of the regional cooperative rescue model implemented on the length of time from first medical contact (FMC) to balloon dilation (B),economic expense and prognosis in patients with acute coronary syndrome (ACS).Methods Patients with ACS (including ST-segment elevation and non-ST-segment elevation) selected from other hospitals within 24 hours after onset were treated with emergency percutaneous coronary intervention.Patients were divided into two groups, regional cooperative rescue group and control group without the regional cooperative rescue model approved.The lengths of FMC-to-B time and Door-to-B time (from arrival at emergency department or OPD to balloon dilation),time required for patients referred to our hospital,cardiac function,averaged hospital costs,average hospital stay,percentage of medication used and a major adverse cardiac event (MACE) were analyzed.Results Mean FMC-to-B time,Door-to-B time,referral time and time consumed to obtain informed consent were significantly shorter [(106±33) min,(31 ±8) min,(62 ±18,8 ±3) min] vs.[(231 ±35) min,(109 ±26) min,(98 ±31) min,(28 ±11) min,respectively] by implementing the regional cooperative rescue compared with control group,and LVEF was increased,and LVED was deceased inregional cooperative rescue group.The mean costs [(44 123.0 ±3 427.0) yuan vs.(51 587.0 ±5 621.0)] yuan,days of hospital stay [(8.7 ±4.1) vs.(13.2 ±6.4)] and percentage of medication used were significantly decreased in the regional cooperative rescue group.The incidence of MACE inregional cooperative rescue group was 6.2％,whereas the incidence in control group was 16.8％.Conclusions The regional cooperative rescue model can improve the prognosis and decrease the FMC-to-B time,the rate of MACE and financial burden in patients with ACS.
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<p><b>OBJECTIVE</b>To investigate the association between CD147 expression and its untranslated regions 3'UTR rs8259 T/A polymorphism and acute coronary syndrome (ACS).</p><p><b>METHODS</b>The genotypes of CD147 were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods in 182 ACS patients and 328 healthy controls. The plasma level of CD147 was determined by enzyme-linked immunosorbent assay (ELISA). CD147 mRNA and protein expression was detected by real-time fluorescent quantitative PCR (RT-qPCR) and Western blot.</p><p><b>RESULTS</b>The plasma CD147 level obtained from radial artery in ACS patients ((3.63 ± 0.70) pg/L) was significantly higher than in control ((2.45 ± 0.27) pg/L, P < 0.05), and highest in plasma obtained from the coronary artery ((4.28 ± 1.03) pg/L, P < 0.05) in ACS patients. Furthermore, the plasma CD147 level was higher in the ACS patients with rs8259 AA genotype than in the ACS patients with rs8259 TT genotype ((4.08 ± 0.41) pg/L vs. (3.05 ± 0.79) pg/L in radial artery and (5.29 ± 0.62) pg/L vs. (3.13 ± 0.52) pg/L in coronary artery, both P < 0.05). There are an enhanced expression of CD147 mRNA (2.45 times higher than control) and protein (3.66 ± 1.56 vs. 1.81 ± 1.29) in PBMCs from ACS patients than that from controls (both P < 0.05). The PBMCs CD147 mRNA and protein expression level were significantly higher in ACS patients with rs8259 AA genotype (mRNA:2.45 ± 0.35, protein:1.63 ± 0.16) compared to ACS patients with rs8259 TT genotype (mRNA:1.69 ± 0.15, protein: 0.88 ± 0.16, both P < 0.05). Multiple logistic analysis showed that CD147 T allele (AT+TT) was a protective factor to ACS (OR = 0.667, 95% CI 0.507-0.879, P < 0.05).</p><p><b>CONCLUSIONS</b>The over-expression of CD147 is involved in the pathogenesis of ACS. The CD147 3'UTR rs8259 T allele may be a protective factor for ACS, its polymorphism can affect the CD147 protein expression in ACS patients.</p>
Subject(s)
Humans , Acute Coronary Syndrome , Genetics , Alleles , Basigin , Genetics , Case-Control Studies , Genotype , Leukocytes, Mononuclear , Polymorphism, Genetic , Polymorphism, Restriction Fragment LengthABSTRACT
<p><b>OBJECTIVE</b>To evaluate the effect of new regional cooperative rescue model on the first medical contact-to-balloon time and outcome in patients with ST-elevation myocardial infarction.</p><p><b>METHOD</b>Patients with acute myocardial infraction (AMI) and onset time within 24 hours transferred from other hospitals to our clinic and underwent emergent percutaneous coronary intervention (PCI) between January 2010 and January 2013 were included in this study. Patients were divided into two groups: regional cooperative treatment group (n = 230) and control group (n = 168) according to whether the first contact clinic belongs to the regional cooperative rescue model or not. The first medical contact to balloon (FMC-to-B) time, door to balloon (D-to-B) time, referral time, cardiac function, mean cost, days of hospitalization, and major adverse cardiac event (MACE) during the 6 months follow up were compared.</p><p><b>RESULTS</b>Mean FMC-to-B time, D-to-B time and referral time were significantly decreased from (212 ± 37), (107 ± 18), (103 ± 23) min (control group) to (98 ± 23), (25 ± 7), (62 ± 12) min respectively in regional cooperative treatment group. Mean medical cost (42 221 (23 184, 77 768) RMB vs. 49 654 (25 126, 122 433) RMB) and days of hospitalization (7 (5, 13) days vs. 10 (6, 20) days) were also significantly lower in regional cooperative treatment group than in control group. At 6 months follow up, LVEF was significantly higher(54.9% ± 8.6% vs. 48.9% ± 9.1%, P = 0.01), LVEDD ((48.9 ± 5.7)mm vs.(51.4 ± 6.0) mm, P < 0.01) as well as MACE rate (7.4% (17/230) vs. 17.9% (30/168) , P < 0.05) were significantly lower in regional cooperative treatment group than in control group.</p><p><b>CONCLUSION</b>The regional cooperative rescue model can decrease the FMC-to-B time, improve cardiac function, and reduce both patients' financial burden and MACE in patients with acute myocardial infarction.</p>
Subject(s)
Humans , Angioplasty, Balloon, Coronary , Hospitalization , Myocardial Infarction , Therapeutics , Percutaneous Coronary Intervention , Regional Health Planning , Time FactorsABSTRACT
Objective To investigate the correlation between the CD40 gene( - 1 C/T)single nucleotide polymorphism(SNP) and acute coronary syndromes(ACS), and the expression of CD40 on platelets. Method A total of 562 patients with ACS canfirmed by coronary angiography were divided into 3 groups according to the clinical characteristics, namely ACS patients( n = 210), stable angina(SA) patients( n = 189) and control group( n = 163).ACS was defined as ischemic chest pain at rest resulting in admission to hospital and > 50% stenosis in a major coronary artery with or without a rise in troponin Ⅰ. SA was defined as stable effort-related angina without change in angina pattern in 3 months. Patients with infection, tumor, or liver or kidney disease were excluded The gene polymorphism was measured by the polymerase chain reaction and restriction fragment length polymorphism(PCR-RFIP) and identiffed by sequencing. The expression of CD40 on platelets was detected by flow cytometry. The frequency, distribution of genotypes was compared using cross-tabulation and standard X~2 test. Result The CC genotype(31% ) and C allele of frequency(57.9%)of CD40 gene in ACS patients were significantly higher than those in SA(15.9%, 43.1% ) and control groups( 16.1%, 42.6% ). No significant difference of the genotypes or allele frequencies was found between SA and control group(X~2 = 0.053, P = 0.974;X~2 = 0.017, P = 0.897). 1he expression of CD40 on platelets in patients with C alleles carries was significandy higher than that of T allele carries in each group( P <0.0001). Conclusions CD40- 1C/T polymorphism was associated with ACS in Chinese Han nationallity.
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BACKGROUND: Non-invasive vascular ultrasound is used to assess in tima-media thickness (IMT) and atheromatous plaque, of which the result is better correlated to that of pathological examination. OBJECTIVE: Non-invasive vascular ultrasound was used to assess the hemodynamical changes of external and internal carotid in hypertensive elders, so as to investigate the relationship between pulse pressures, left ventricular mass index (LVMI), vascular resistance index (VRI) and shearing ratio. DESIGN: Randomized, controlled, correlative analyzedstudy. SETTING: Ultrasound Department of Affiliated Hospital of Jiangshu University. PARTICIPANTS: Totally 54 hypertensive elders, including 33 males and 21 females, were recruited from the Ultrasound Department of Affiliated Hospital of Jiangshu University, from September 30th 2000 to March 30th 2002. Meanwhile 26 normotensive elders (systolic pressure ＜ 130 mm Hg and/or diastolic pressure ＜ 85 mm Hg), including 16 males and 10 females, were selected as controls, METHODS: Color duplex blood flow imaging technique was used to assess the left ventricular end-diastolic diameter (LVEDD) and the interventricular septum thickness (IVST), so as to calculate the LVMI. The insiddiameter of bilateral common carotid artery (CCA) and internal carotid,IMT, the systolic peak flow velocity and resistance index were measured in order to calculate the vascular shearing ratio and atheromatous plaque index,moreover arterial pressure, LVMI, as well as resistance index and shearing ratio were subjected to linear correlative analysis,of which the results were compared with that of control group. MAIN OUTCOME MEASURES: ① Pulse pressure, LVMI and atheromatous plaque index of CCA. ② The inside diameter of bilateral CCA and internal carotid, IMT and the systolic peak flow velocity and resistance index were measured in order to calculate the atheromatous plaque index and maximum vascular shearing ratio. RESULTS: According to the experimental objective,data of all 80 cases were analyzed without loss. ① The pulse pressure and LVMI, the incidence of atheromatous plaque and plaque index of CCA in hypertensive elder group were obviously higher than that of control group [(64.36±7.44) mm Hg,(45.78±8.72) mm Hg; (131.47±18.73) g/m2, (83.28±12.34) g/m2; 5.62 ±1.78,3.44±1.10; 66.7％, 23.1％, (x2=13.37, t=5.730-13.234, P ＜ 0.01)]. ② The inside diameter of bilateral CCA and internal carotid, IMT and resistance index in hypertensive elders were significantly higher than that of control group [CCA: (8.96±1.20) mm, (8.08±0.96) mm; (1.28±0.88) mm, (0.91±0.17) mm;0.75±0.05, 0.69±0.06; ICA: (6.82±1.12) mm, (5.72±0.92) mm; (1.06±0.17) mm,(0.76±0.15) mm; 0.70±0.07, 0.64±0.06, t=2.872-10.850, P ＜ 0.01], but the systolic peak flow velocity and shearing ratio were markedly lower than that of control group [CCA: (47.44±12.85) cm/s, (60.20±14.32) cm/s;(225.78 ±95.76)/s, (300.26 ±110.42)/s. ICA: (41.22 ±10.18) cm/s,(48.64±11.58) cm/s; (268.54±112.56)/s, (324.52±121.42)/s, t=2.872-10.850, P ＜ 0.01]. ③ The pulse pressure, LVMI, the resistance index of CCA and ICA and shearing ratio were linearly correlated (r=0.30-0.36, P ＜ 0.05). CONCLUSION: Pulse pressure and LVMI were found increased in hypertensive elders, together with carotid vessels dilated, IMT and resistance index increased, as well as shearing ratio decreased, atheromatous plaque formed, vascular stiffness increased, which resulted in consequently decrease of vascular compliance.