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The dynamic characteristics of the area of the atrial septal defect (ASD) were evaluated using the technique of real-time three-dimensional echocardiography (RT 3DE),the potential factors re sponsible for the dynamic characteristics of the area of ASD were observed,and the overall and local volume and functions of the patients with ASD were measured,RT 3DE was performed on the 27 normal controls and 28 patients with ASD.Based on the three-dimensional data workstations,the area of ASD was measured at P wave vertex,R wave vertex,T wave starting point,and T wave terminal point and in the T-P section.The right atrial volume in the same time phase of the cardiac cycle and the motion displacement distance of the tricuspid annulus in the corresponding period were measured.The measured value of the area of ASD was analyzed.The changes in the right atrial volume and the motion displacement distance of the tricuspid annulus in the normal control group and the ASD group were compared.The right ventricular ejection fractions in the normal control group and the ASD group were compared using the RT 3DE long-axis eight-plane (LA 8-plane) method.Real-time three-dimensional volume imaging was performed in the normal control group and ASD group (n=30).The right ventricular inflow tract,outflow tract,cardiac apex muscular trabecula dilatation,end-systolic volume,overall dilatation,end-systolic volume,and appropriate local and overall ejection fractions in both two groups were measured with the four-dimensional right ventricular quantitative analysis method (4D RVQ) and compared.The overall right ventricular volume and the ejection fraction measured by the LA 8-plane method and 4D RVQ were subjected to a related analysis.Dynamic changes occurred to the area of ASD in the cardiac cycle.The rules for dynamic changes in the area of ASD and the rules for changes in the right atrial volume in the cardiac cycle were consistent.The maximum value of the changes in the right atrial volume occurred in the end-systolic period when the peak of the curve appeared.The minimum value of the changes occurred in the end-systolic period and was located at the lowest point of the volume variation curve.The area variation curve for ASD and the motion variation curve for the tricuspid annulus in the cardiac cycle were the same.The displacement of the tricuspid annulus exhibited directionality.The measured values of the area of ASD at P wave vertex,R wave vertex,T wave starting point,T wave terminal point and in the T-P section were properly correlated with the right atrial volume (P＜0.001).The area of ASD and the motion displacement distance of the tricuspid annulus were negatively correlated (P＜0.05).The right atrial volumes in the ASD group in the cardiac cycle in various time phases increased significantly as compared with those in the normal control group (P=0.0001).The motion displacement distance of the tricuspid annulus decreased significantly in the ASD group as compared with that in the normal control group (P=0.043).The right ventricular ejection fraction in the ASD group was lower than that in the normal control group (P=0.032).The ejection fraction of the cardiac apex trabecula of the ASD patients was significantly lower than the ejection fractions of the right ventricular outflow tract and inflow tract and overall ejection fraction.The difference was statistically significant (P=0.005).The right ventricular local and overall dilatation and end-systolic volumes in the ASD group increased significantly as compared with those in the normal control group (P=0.031).The aRVEF and the overall ejection fraction decreased in the ASD group as compared with those in the normal control group (P=0.0005).The dynamic changes in the area of ASD and the motion curves for the right atrial volume and tricuspid annulus have the same dynamic characteristics.RT 3DE can be used to accurately evaluate the local and overall volume and functions of the right ventricle.The local and overall volume loads of the right ventricle in the ASD patients increase significantly as compared with those of the normal people.The right ventricular cardiac apex and the overall systolic function decrease.
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OBJECTIVES: To explore the effects of serum from patients with ankylosing spondylitis on the canonical Wnt/β-catenin pathway and to assess whether the serum has an osteogenic effect in MG63 cells. METHODS: MG63 cells were cultured with serum from 45 ankylosing spondylitis patients, 30 healthy controls, or 45 rheumatoid arthritis patients. The relative PPARD, fra-1, MMP7, OPG and RANKL mRNA levels were measured using quantitative real-time polymerase chain reaction. Associations between gene expression and patient demographics and clinical assessments were then analyzed. RESULTS: MG63 cells treated with serum from ankylosing spondylitis patients had higher PPARD, fra-1, MMP7 and OPG gene expression than did cells treated with serum from controls or rheumatoid arthritis patients (all p<0.05). RANKL expression was higher in MG63 cells treated with serum from patients with ankylosing spondylitis or rheumatoid arthritis than in those treated with serum from controls (both p<0.05). The OPG/RANKL ratio was also higher in MG63 cells treated with serum from ankylosing spondylitis patients than in those treated with serum from controls (p<0.05). No associations were found between the expression of the five genes and the patient demographics and clinical assessments (all p>0.05). CONCLUSIONS : Serum from ankylosing spondylitis patients increases PPARD, fra-1, MMP7, OPG and RANKL expression and the OPG/RANKL ratio in MG63 cells; these effects may be due to the stimulatory effect of the serum on the Wnt pathway.
Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Artrite Reumatoide/sangue , Osteoblastos/metabolismo , Soro , Espondilite Anquilosante/sangue , Células Cultivadas , Meios de Cultura , Citocinas/metabolismo , Expressão Gênica , /metabolismo , PPAR delta/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ligante RANK/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Via de Sinalização Wnt/genética , beta Catenina/genéticaRESUMO
This study aimed to examine the preparation of cationic lipid microbubble (CLM),and evaluate its physical and chemical properties and toxicity,measure the gene transfection efficiency by ultrasound triggered microbobble destruction (UTMD) in combination with CLM.The CLM was prepared by the method of the thin film hydration,and its morphology was observed under the electron microscopy at 1st,3rd,7th,10th,and 14th day after preparation,respectively.The size,Zeta potential and stability of CLM were tested.The acute toxicity of CLM was assessed.The green fluorescent protein gene (EGFP) transfection efficiency was evaluated.The experiment grouping was as follows:naked plasmid group (P group),ultrasonic irradiation plus naked plasmid group (P-US group),naked plasmid plus CLM group (P-CLM group),naked plasmid plus ultrasound and CLM group (UTMD group).The expression of EGFP was detected by fluorescent microscopy and flow cytometry.The results showed that CLMs were spherical in shape,with the similar size and good distribution degree under the light and electron microscopies.The size of CLMs was varied from 250.4±88.3 to 399.0±99.8 nm and the Zeta potential of CLMs from 18.80±4.97 to 20.1 ±3.1 mV.The EGFP expression was the strongest in the UTMD group,followed by the P-CLM group,P-US group and P group.Flow eytometry results were consistent with those of fluorescent microscopy.The transfection efficiency was substantially increased in the P-US group,P-CLM group and UTMD group as compared with that in the P group,almost 7 times,10 times and 30 times higher than that in the P group respectively.It is suggested that CLMs prepared by the method of thin film hydration are uniform in diameter,and proved non-toxic.UTMD combined with CLM can significantly increase the transfection efficiency of EGFP to targeted cells.
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To explore the protective effect of sodium tanshinone Ⅱ A sulfonate (STS) on microcirculatory disturbance of small intestine in rats with sepsis,and the possible mechanism,a rat model of sepsis was induced by cecal ligation and puncture (CLP).Rats were randomly divided into 3 groups:sham operated group (S),sepsis group (CLP) and STS treatment group (STS).STS (1 mg/kg) was slowly injected through the right external jugular vein after CLP.The histopathologic changes in the intestinal tissue and changes of mesenteric microcirculation were observed.The levels of tumor necrosis factor-α(TNF-α) in the intestinal tissue were determined by using enzyme-linked immunoabsorbent assay (ELISA).The expression of intercellular adhesion molecule-1 (ICAM-1) in the intestinal tissue was detected by using immunohistochemisty and Western blot,that of nuclear factor κB (NF-κB) and tissue factor (TF) by using Western blot,and the levels of NF-κB mRNA expression by using RT-PCR respectively.The microcirculatory disturbance of the intestine was aggravated after CLP.The injury of the intestinal tissues was obviously aggravated in CLP group as compared with S group.The expression levels of NF-κB p65,ICAM-1,TF and TNF-α were upregulaed after CLP (P＜0.01).STS post-treatment could ameliorate the microcirculatory disturbance,attenuate the injury of the intestinal tissues induced by CLP,and decrease the levels of NF-κB,ICAM-1,TF and TNF-α (P＜0.01).It is suggested that STS can ameliorate the microcirculatory disturbance of the small intestine in rats with sepsis,and the mechanism may be associated with the inhibition of inflammatory responses and amelioration of coagulation abnormality.
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This study examined the wave intensity (WI) of the carotid artery in patients with hyperthyroid in order to assess the hemodynamic changes of hyperthyroid patients. A total of 86 hyperthyroid patients without cardiac morphological changes and arrhythmia, and 80 healthy control subjects were enrolled in the study. Right common carotid artery (RCCA) was selected for ultrasonic imaging to obtain WI indices, including amplitude of the peak during early systole (W1), amplitude of the peak during late systole (W2), area of the negative wave during mid-systole (NA), interval between R wave of electrocardiogram and W1 (R-1 st), interval between W1 and W2 (1 st-2nd). The levels of serum thyroid hormones, consisting of free triiodothyronine (FT3), free thyroxin (FT4) and thyroid stimulating hormone (TSH), were measured in hyperthyroid patients. Echocardiographic indices including left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS) were determined in each subject. The results showed that the WI, W2, NA, and (lst-2nd×HR) in hyperthyroid patients were significantly higher than those in healthy controls. There was no significant difference in LVEF and LVFS between the two groups. FT3 was correlated with WI, W2, NA,(lst-2nd×HR), pulse pressure (PP) and heart rate (HR) in hyperthyroid patients. Several abnormal waves on WI curves were present in 19 hyperthyroid patients during mid-systole. It was concluded that WI technique may prove a real-time, noninvasive, sensitive and convenient tool for assessing the cardiac function and hemodynamic alterations in hyperthyroid patients.
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Quantification of right ventricular(RV)volume and function remains a challenge because of RV complex geometry by conventional echocardiography.The purpose of this study was to assess RV global longitudinal function in patients with tetralogy of Fallot(TOF)by 2-dimensional ultrasound speckle tracking imaging(STI).Thirty-eight patients with TOF were enrolled in this study and divided into child group(n=25)and adult group(n=13)according to age.Thirty-eight age-and sex-matched normal subjects were selected as control groups including child control group(n=25)and adult control group(n=13).RV global longitudinal peak systolic strain(GLS),strain rate(GLSPs),early diastolic strain rate(GLSRe)and late diastolic strain rate(GLSRa)were measured in all subjects by STI from the apical 4-chamber view and compared between groups.Furthermore,the main factors affecting the RV global longitudinal functional parameters were assessed.Compared with those in controls,RV GLS,GLSRs and GLSPe were significantly reduced in patients with TOF(P<0.05 for all).RV GLSRs was significantly decreased in adult patients with TOF as compared with that in child patients(P<0.05).There was no significant difference in RV GLS,GLSRe and GLSRa between child and adult TOF groups(P>0.05).The diameter of right ventricle,main pulmonary artery and ventricular septum defect had correlations with RV GLSRs(r1=-0.490,r2=0.580,r3=-0.528,respectively,P<0.05 for all).Tricuspid annular plane peak systolic velocity(Sm)was the independent predictor of RV global strain and strain rate(β1=0.355,P1=0.031,β2=0.307,P2=0.021).RV global longitudinal function is decreased in patients with TOF,especially in adult patients.STI is a sensitive and accurate technique in RV global functional assessment.
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This study evaluated the change in regional left ventricular myocardial function in rats following acute occlusion of the left anterior descending coronary artery (LAD) by using two-dimensional speckle tracking imaging (2D-STI). Sixty Wistar rats were randomly divided into two groups,a myocardial infarction (MI) group,in which 50 rats were subjected to LAD occlusion for 30-45 min,and a sham-operated (SHAM) group that contained 10 rats serving as control. Echocardiography was performed at baseline and 1,4 and 8 week(s) after the operation. High frequency two-dimensional images of left ventricular short axis at papillary muscle level were recorded. Peak systolic radial strain (PRS) and circumferential strain (PCS) were measured in the mid-ventricle in short-axis view by using EchoPAC workstation. Left ventrieular internal diameter at diastole (LVIDd)and systole (LVIDs),fractional shortening (FS),ejection fraction (EF) and left ventricular mass (LVM)were measured by anatomical M-model echocardiography. Infarct size was measured using triphenyl tetrazolium chloride (TTC) staining 1 week and 8 weeks after the operation. Fibrosis of left ventricular myocardium was displayed using Van Gieson staining 1 week after the infarction. In terms of the TTC staining results,the left ventricle fell into three categories:infarcted,peri-infarcted and remote myocardial regions. Compared with those at baseline and in the SHAM group,(1) PRS and PCS in the infarcted,peri-infarcted and remote myocardial regions were significantly decreased in the MI group within 1 week after the operation (P＜0.05) and the low levels lasted 8 weeks;(2) Compared with those at baseline,LVIDd,LVIDs,FS,EF and LVM in the MI group showed no significant difference 1 week after the operation (P＞0.05). However,LVIDd,LVIDs and LVM were increased significantly 4 and 8 weeks after the operation (P＜0.05),and FS and EF were decreased substantially (P＜0.05). Van Gieson staining showed that fibrosis developed in all the three myocardial regions to varying degrees. It is concluded that 2D-STI is non-invasive and can be used to assess regional function of myoeardium with different blood supply in rats following acute occlusion of the LAD,and can be used as a sensitive and reliable means to follow up the process of left ventricular remodeling.
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Stereoscopic three-dimensional echocardiography(S-3DE) is a novel displaying technology based on real-time 3-dimensional echocardiography (RT-3DE). Our study was to evaluate the feasibility and efficiency of S-3DE in the diagnosis of atrial septal defect (ASD) and its use in the guidance for transcatheter ASD occlusion. Twelve patients with secundum ASD underwent RT-3DE examination and 9 of the 12 were subjected to transcatheter closure of ASD. Stereoscopic vision was generated with a high-performance volume renderer with red-green stereoscopic glasses. S-3DE was compared with standard RT-3D display for the assessment of the shape,size,and the surrounding tissues of ASD and for the guidance of ASD occlusion. The appearance rate of coronary sinus and the mean formation time of the IVC,SVC were compared. Our results showed that S-3DE could measure the diameter of ASD accurately and there was no significant difference in the measurements between S-3DE and standard 3D display (2.89±0.73 cm vs 2.85±0.72 cm,P＞0.05;r=0.96,P＜0.05). The appearance of coronary sinus for S-3DE was higher as compared with the standard 3D display (93.3% vs 100%). The mean time of the IVC,SVC for S-3DE monitor was slightly shorter than that of the standard 3D display (11.0±3.8 s vs 10.3±3.6 s,P＞0.05). The mean completion time of interventional procedure was shortened with S-3DE display as compared with standard 3D display (17.3±3.1min vs 23.0±3.9 min,P＜0.05). Stereoscopic three-dimensional echocardiography could improve the visualization of three-dimensional echocardiography,facilitate the identification of the adjacent structures,decrease the time required for interventional manipulation. It may be a feasible,safe,and efficient tool for guiding transcatheter septal occlusion or the surgical interventions.
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To evaluate the feasibility of real-time myocardial contrast echocardiography (RTMCE) by quantitative analysis of myocardial perfusion in rabbits,transthoracic RTMCE was performed in 10 healthy rabbits by using continuous infusion of Sono Vue into the auricular vein. The short axis view at the papillary muscle level was obtained. The duration of the time that the contrast took to appear in right heart,left heart and myocardium was recorded. The regional myocardial signal intensity (SI) versus refilling time plots were fitted to an exponential function:y(t) =A(1-e-β(t-t0)) + C,where y is SI at any given time,A is the SI plateau that reflects myocardial blood volume,and β is the slope of the refilling curve that reflects myocardial microbubble velocity. The A,β and A×β values at different infusion rate of SonoVue were analyzed and the A,β and A×β values in each segment in the short axis view at the papillary muscle level were compared. All the animal experiments were successful and high-quality images were obtained. The best intravenous infusion rate for Sono Vue was 30 mL/h. The contrast appeared in fight heart,left heart and myocardium at 7.5±2.2 s,9.1±2.4 s and 12.2±1.6 s respectively. After 16.6±2.3s,myocardial opacification reached a steady state. The mean A,β and A×β value in the short axis view at the papillary muscle level were 9.8±3.0 dB,1.4±0.5 s 1 and 13.5±3.6 dB×s-1 respectively.A,β and A×β values showed no significant differences among 6 segments. It was suggested that RTMCE was feasible for quantitative analysis of myocardial perfusion in rabbits. It provides a non-invasive method to evaluate the myocardial perfusion in rabbit disease models.
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Left ventricular remodeling index (LVRI) was assessed in patients with hypertensive heart disease (HHD) and coronary artery disease (CAD) by real-time three-dimensional echocardi-ography (RT3DE).RT3DE data of 18 patients with HHD,20 patients with CAD and 22 normal con-trols (NC) were acquired.Left ventricular end-diastolic volume (EDV) and left ventricular end-diastolic epicardial volume (EDVepi) were detected by RT3DE and two-dimensional echocar-diography Simpson biplane method (2DE).LVRI (left ventricular mass/EDV) was calculated and compared.The results showed that LVRI measurements detected by RT3DE and 2DE showed sig-nificant differences inter-groups (P<0.01).There was no significant difference in NC group (P>0.05),but significant difference in HHD and CAD intra-group (P<0.05).There was good positive correla-tions between LVRI detected by RT3DE and 2DE in NC and HHD groups (r=0.69,P<0.01; r=0.68,P<0.01),but no significant correlation in CAD group (r=0.30,P>0.05).It was concluded that LVRI derived from RT3DE as a new index for evaluating left ventricular remodeling can provide more su-periority to LVRI derived from 2DE.
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We studied the wall motion characteristics of the ascending aorta by velocity vector im-aging (VVI) in primary hypertension patients.The ascending aortas both in 30 patients with primary hypertension and 30 normal controls were examined by Acuson sequoia 512 equiped with VVI.The maximum velocity (Vs,Ve) of every point on the anterior wall of ascending aorta both in systole and diastole was measured.The aortic diameter was wider in the hypertension patients than that in the healthy subjects (P<0.05).The movement amplitude of the anterior wall of the ascending aorta in long axis view in the hypertension patients was lower than that in the healthy subjects (P<0.05).The motion and time to peak in systole of each point of the ascending aorta in the healthy subjects had no significant difference (P>0.05).The velocity curves of the anterior wall of ascending aorta both in the hypertension and healthy subjects were regular,and the curve in systole was named S wave and that in diastole named E wave.The velocity of S wave and E wave was slower in the hypertension pa-tients than that in the healthy subjects (P<0.05).The time to peak of S wave on the anterior wall of ascending aorta in systole was shorter in the hypertension patients than in the healthy subjects (P<0.05).VVI could be used to accurately and directly observe the movement character of the as-cending aorta walls,which would help us understand the elasticity of great arteries in patients with hypertension.
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Summary: The aim of present study was to evaluate the feasibility and efficiency of enhanced green fluorescent protein (EGFP) gene delivery to myocardium in vivo by ultrasound targeted microbubble destruction (UTMD) and polyethylenimine (PEI). SonoVue/DNA and PEI/DNA/SonoVue complexes were prepared. Gel electrophoresis analysis was performed to determine the structural integrity of plasmid DNA or PEI/DNA after UTMD. Solutions of plasmid DNA, SonoVue/DNA, PEI/DNA complexes or PEI/DNA/SonoVue complexes were respectively transduced into BALB/c mice hearts by means of transthoracic ultrasound irradiation. Mice undergoing PBS injection, plasmid injection or PEI/DNA complexes injection without ultrasound irradiation served as controls. Gene expression in myocardium was detected 4 days after treatment. Cryosections and histological examinations were conducted. Electrophoresis gel assay showed no damage to DNA or PEI/DNA complexes after UTMD. When the heart was not exposed to ultrasound, the expression of EGFP was observed in the subendocardial myocardium obviously. The strongest expression was detected in the anterior wall of the left ventricle when the heart was exposed to ultrasound alone. Injection of PEI/DNA complexes and UTMD resulted in the highest transfection efficiency and the distributional difference of EGFP was not obvious. No tissue damage was seen histologically. In conclusion, a combination of UTMD and PEI was highly effective in transfecting mice hearts without causing any apparently adverse effect. It provides an alternative to current clinical gene therapy and opens a new concept of non-viral gene delivery for the treatment of cardiac disease.