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Objective:To explore the role of activated CD4 + T cells in cardiac remodeling after myocardial infarction (MI). Methods:① Experiment in vitro: naive CD4 + T cells were isolated in mouse spleen, and then stimulated with plate-bound anti-CD3 and anti-CD28 for 48 hours. Exosomes isolated from the supernatant of activated CD4 + T cells were incubated with cardiac fibroblasts (CFs) for 48 hours, and then the ability of CFs proliferation, migration and differentiation were detected by cell counting kit-8 (CCK-8) assay, Transwell assay, and immunofluorescence assay. ② Experiment in vivo: 40 male C57 mice were divided into 4 groups according to random number table method, including control group (Ctrl group), sham operation group (Sham group), MI group, and exosome treatment group (MI+Exo group), with 10 in each group. The mice model of MI was established by ligating the left anterior descending coronary artery. In MI+Exo group, 40 μg/d exosomes were injected intravenously into the tail after modeling. Cardiac function and cardiac fibrosis post-MI were assessed by echocardiography and quantitative polymerase chain reaction (qPCR) at 4th week. Results:① In vitro: exosomes derived from activated CD4 + T cells significantly promote CFs proliferation, migration and differentiation [proliferation ability ( A value): 0.31±0.01 vs. 0.21±0.01, migration capability (cells/MP): 79.20±3.34 vs. 48.80±2.13, differentiation ability (α-smooth muscle actin, α-SMA; fluorescence intensity): 1.56±0.03 vs. 1.00±0.02, all P < 0.05]. ② In vivo: echocardiographic analysis showed that exosomes derived from activated CD4 + T cells aggravated the deterioration of cardiac dysfunction post-MI than MI group, as indicated by left ventricular ejection fraction (LVEF) and fractional shortening (FS) decreased significantly [LVEF: 0.185±0.008 vs. 0.257±0.022, FS: (9.72±1.72)% vs. (14.08±1.08)%, both P < 0.05], left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic diameter (LVESD) increased significantly [LVEDD (mm): 5.43±0.29 vs. 4.62±0.35, LVESD (mm): 4.94±0.12 vs. 3.69±0.29, both P < 0.05]. Additionally, qPCR showed that exosomes derived from activated CD4 + T cells remarkably promoted myocardial fibrosis post-MI than MI group, as indicated by the mRNA expression of α-SMA, collagens (Col1a1, Col3a1) in MI+Exo group was significantly higher than that in MI group [α-SMA (2 -ΔΔCT): 4.72±0.89 vs. 3.58±0.78, Col1a1 (2 -ΔΔCT): 6.59±0.56 vs. 4.23±0.42, Col3a1 (2 -ΔΔCT): 13.40±1.03 vs.4.96±0.36, all P < 0.05]. Conclusion:Activated CD4 + T cells promote cardiac remodeling following MI through transferring exosomes to CFs.
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AIM:To investigate the effects of leptin on the expression of bile salt export pump ( BSEP) and signaling pathway in human hepatocellular carcinoma cell line HepG2.METHODS: HepG2 cells were cultured in vitro. Leptin at concentrations of 10 -8 , 10 -7 and 10 -6 mol/L was used as a stimulating factor.The protein levels of adenosine monophosphate-activated protein kinase alpha subunit (AMPKa), phosphorylated AMPKa (p-AMPKa) and BSEP in the HepG2 cells at 24 h, 48 h and 72 h were detected by Western blotting.The optimal culture time and leptin concentration were selected, and compound C at concentration of 10 μmol/L was added to this group.The protein expression of BSEP was detected by Western blotting.RESULTS:Intervention of HepG2 cells with leptin for 72 h increased the protein ex-pression of AMPKa gradually in a concentration-dependent manner, and leptin at concentration of 10 -6 mol/L induced the strongest AMPKa expression ( P<0.01 ) .Intervention of HepG2 cells with leptin for 24 h increased the phosphorylation level of AMPKa gradually in a dose-dependent manner (P<0.01).The effect of leptin on the increase in the protein ex-pression of p-AMPKa was also in a time-dependent manner ( P<0.01) .After intervention with different concentrations of leptin for 24 h, the protein expression of BSEP in the HepG2 cells was gradually increased by the stimulation of leptin in a concentration-and time-dependent manner (P<0.01).Compared with NC group, the protein expression of BSEP in 10 -6 mol/L leptin group and 10 -6 mol/L leptin+10μmol/L compound C group was increased at 72 h (P<0.01), and that in 10-6 mol/L leptin+10 μmol/L compound C group was lower than that in 10 -6 mol/L leptin group at 72 h ( P<0.01 ) . CONCLUSION:Leptin promotes the protein expression of BSEP in HepG2 cells by leptin-AMPK-BSEP signaling path-way.Leptin promotes the increases in AMPKa protein and the level of phosphorylation of AMPKa in HepG2 cells.
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Objective The purpose of this study is to select suitable ages of rats for the CPR ( cardiopulmonary resuscitation) animal model.The neurological function score and subgroups analysis are evaluated in 2 month old and 4 month old animal groups.Methods Based on the evaluation of physiological indexes including ECG, blood pressure and neurological function defect score ( NDS) and subgroup analysis, the stability of CPR rats model was compared between 2 month old and 4 month old animal groups.Results The results showed that, the model rate of the ventricular fibrillation was induced by electrical stimulation , the 4 month old group was 87.5%, significantly higher than the 2 month old group, however, there was no significant difference between the two groups in the mortality rate;For the changes of blood pressure during the process of CA( cardiac arrest) induced by electrical stimulation, the 4 month old group was significantly lower than the 2 month old group (P <0.01); for the NDS at each time point after CPR, there was no significant difference between the two groups; however, the NDS subgroup analysis at different time points showed that there were different degrees of differences between the two age groups ( P <0.05) .Comparing with the 2 month old group, the 4 month old group had a stable process during the animal model preparation, had an obvious cerebral blood hypoperfusion phenomenon and aggravation of brain injury after CPR.Conclusion The 4 month old rats are more suitable for preparation of CPR animal mode , the model rate is high, the brain injury aggravate.It is more suitable evaluation for basic research and treatment of CPR.
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Objective:To explore the effect of intraventricular administration of insulin on pro-apoptotic expression of Caspase-3 mRNA and neuronal apoptosis in hippocampus CA1, and neurological function after rats’ CPR. Methods:Thirty male SD rats were randomly divided into three groups:sham group;the resuscitation with saline-treated group and resuscitation with insulin-treated group. Six minute cardiac arrest was induced by ventricular fibrillation (VF) via pacing electrode placed in rats’ esophagus in saline and insulin group. Results:1. The NDS revealed a clear neurological deficit after reperfusion 24 h, 72 h in insulin and saline group as compare to the sham group, Comparing to the saline group, Insulin could improve the rats’ neurologic function after CPR 24 (insulin vs. saline group, 70(64~72)vs.56(50~58), P<0.001);2. Through TUNEL stain, insulin inhibited apoptosis in CA1 hippocampus as compare to the saline after CPR 7d. (F=5.853, P=0.02) 3.Caspase-3 expression in insulin-treated groups were significantly decreased compared to the saline-treated group after reperfusion 24h and 72h[(70(64~72), 56(50~58);P<0.001]. Conclusion:Intraventricular administration of insulin could inhibit the mRNA expression levels of the apoptotic activities of Caspase-3 after cardiac arrest, prevent neuronal apoptosis in the CA1 hippocampus and improve the rats’ neurological function (at least 24hours) after CPR.
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Adenosine monophosphate-activated protein kinase (AMPK) activation also can inhibit the synthesis of cholesterol and fat.Recent studies have shown that activation of AMPK can also inhibit the synthesis of bile acid and promote bile salt export pump' s generation.All of those illustrated that AMPK played an important role in regulating bile acid metabolism.This article will summarize the AMPK activation pathway,bile acid metabolism,AMPK activity in bile acid synthesis and transfer,and so on.
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Objective To investigate the natural course and treatment of adult liver hemangioma (ALH).Methods The records of ultrasonography of 2422 patients with ALH were retrospectively reviewed.Results The patients were between 16 and 69 years of age (mean age 42,9± 11.4) with a male to female ratio of 1 ∶ 1.02 (1197/1225).The prevalence rate gradually increased with age after 30 years old.The highest prevalence rate occurred between 40-69 years.ALH was most commonly 1.1-3 cm in size,located in right liver with a solitary lesion.ALHs were bigger and were seen earlier in females than males.Concurrent illness was also more common in females than males.1427 patients had 3 to 8 repeat sonographies.At a follow-up of 42.7±9.5 months,1351 patients had no change in the ultrasonographic pattern or number of haemangiomas.Increasing in size of the lesions was demonstrated in 76 patients.154 patients received operation or interventional treatment.Conclusions The prevalence rate of ALH rose with age.Female sex hormones might accelerate ALH enlargement but they did not induce the formation of ALH.Most ALH remained stable in size and in patterns.Attention should be paid to rule out other illnesses in patients with ALH and with symptoms.
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Objective To explore the effects of intraventricular administration of insulin on the expressions of Bcl-2,Bax mRNA and neuronal hippocampus apoptosis in rats after cardiopulmonary resuscitation (CPR).Methods This experiment was implemented in the animal Laboratory center of Xuanwu Hospital of Capital Medical University.Thirty male SD rats were randomly (random number)divided into three groups:control group (n=6),CPR group (n=12),insulin treated group ( n =12).CPR was performed at 6 minutes after ventricular fibrillation induced by transesophageal overdrive pacing.Resuscitation procedures lasted until restoration of spontaneous circulation (ROSC).ROSC was defined as the recovery of the supraventricular heart rates and the increase of mean arterial pressure (MAP) ＞ 60mmHg for more than 10 minutes.Ten minutes after ROSC in rats,12.5 μL ( 1 U) regular insulin was injected into the left ventricle in the insulin group,and 12.5 μL isotonic saline was injected the control and CPR groups at least 10 minutes.Real-time PCR was used to observe the expressions of Bcl-2,Bax mRNA in hippocampus CAI after reperfusion 24 h and 72 h.TUNEL staining was used to observe the neuronal apoptosis in all groups after reperfusion 7 days.Blood glucose was monitored in rats before and after CPR.Results ① The Bcl-2mRNA in insulin groups were significantly higher than those in the CPR group after 24 h and 72 h (P ＜0.01 ).The expression of Bcl-2 mRNA in 24 h insulin group were significantly higher than those in 72 h insulin group ( P ＜ 0.01 ) ; There were no significantly different in the Bax mRNA between insulin groups and the CPR and the control group after 24 h and 72 h ( P ＞ 0.05 ) ; ②After CPR 7 d,the apoptotic neurons of hippocampal CA1 area in the CPR group ( 124.75 ± 17.35 ) were significantly higher than those in the control group (5.12 ± 3.26) ( P ＜ 0.01 ) and the insulin group (92.79 ± 7.35 )(P ＜0.01 ); the apoptotic neurons in the insulin group were higher than those in the control group (P ＜0.0l ),and the differences were statistically significant.③There were no significant difference in venous blood glucose in the CPR and insulin groups (P ＞ 0.05).Conclusions Insulin may regulate Bcl-2mRNA expression in hippocampus,inhibit neuronal apoptosis and protect neurons after CPR in rats.