[Antiviral effect of hepatitis B virus S gene-specific anti-gene locked nucleic acid in transgenic mice].

Objective: To investigate the antiviral effect of hepatitis B virus (HBV) S gene-specific anti-gene locked nucleic acid (LNA) in transgenic mice. Methods: A total of 30 HBV transgenic mice were randomly divided into blank control group (5% glucose + liposome), unrelated sequence control group, lamivudine control group, antisense LNA control group, and anti-gene LNA group, with 6 mice in each group. The mice in the lamivudine group were given lamivudine by gavage, and LNA was injected via the caudal vein. Quantitative real-time PCR was used to measure serum HBV DNA, ELISA was used to measure serum HBsAg, RT-PCR was used to measure HBV S mRNA level in the liver, and immunohistochemistry was used to measure the level of HBsAg in hepatocytes. Results: At 3, 5, and 7 days after treatment, there were significant changes in the inhibition rates of HBV DNA (37.18%, 50.27%, and 61.46%, respectively) and HBsAg (30.17%, 44.00%, and 57.76%, respectively) achieved by anti-gene LNA (P < 0.01), and there were significant differences between the anti-gene LNA group and the other four control groups (P < 0.05). In the anti-gene LNA group, the relative mRNA expression of HBV S gene was 0.33 and the percentage of HBsAg-positive hepatocytes was 31%, which were significantly different from these two indices in the control groups (P < 0.05). There were no abnormal changes in liver/renal biochemical parameters and HE staining results. Conclusion: Anti-gene LNA targeting at HBV S gene has a strong antiviral effect in transgenic mice, which provides theoretical and experimental bases for gene therapy for HBV.

Antiviral effect of hepatitis B virus S/C gene loci antisense locked nucleic acid on transgenic mice in vivo.

We investigated the effects of hepatitis B virus (HBV) S/C double gene loci antisense locked nucleic acid on replication and expression of HBV in hepatitis transgenic mice. HBV mice (N = 30) were randomly divided into five groups of six mice: 5% glucose solution control, empty liposome control, single-target S, single-target C, and dual-target SC groups. An antisense locked nucleic acid fragment was injected into the mice. Serum HBsAg, serum HBV DNA, HBV C-mRNA expression in liver tissue, HbsAg and HbcAg expression in hepatocytes, serum albumin, alanine transaminase (ALT), urea nitrogen, and creatinine were detected. Liver and kidney sections were examined for the effects of antisense locked nucleic acid. The expression of HBsAg was markedly inhibited; the inhibition rates of the S, C, and SC target groups were 36.63, 31.50, and 54.87%, respectively; the replication of HBV DNA was also inhibited: 23.97, 21.13, and 35.83%, respectively. After injection at 1, 3, and 5 days, the corresponding rates for HBsAg inhibition were 14.40, 25.61, and 31.33%, and for HBV DNA inhibition they were 11.04, 19.24, and 24.13%. Compared with the control group, the differences in serum albumin, ALT, urea nitrogen, and creatinine in each group were not statistically significant, and the number of HbsAg- and HBcAg-positive cells in the mouse liver was significantly reduced. The liver and kidney tissues were normal. The gene therapy had significant inhibitory effects on the replication and expression of HBV in transgenic mice, and double-gene targeting was better than single-gene targeting.

Blocking the expression of the hepatitis B virus S gene in hepatocellular carcinoma cell lines with an anti-gene locked nucleic acid in vitro.

The aim of this study was to investigate the effects of inhibition of the hepatitis B virus (HBV) S gene by polypurine region locked nucleic acid on viral replication in cells. We designed and synthesized a locked nucleic acid, phosphorothioate oligonucleotides, unmodified oligonucleotides, and unrelated control sequence for the hepatitis B virus S gene polypurine region. HepG2.2.15 cells were transfected by cationic liposome, and fluorescence quantitative polymerase chain reaction technology (PCR) and time-resolved fluoroimmunoassay technology was utilized to monitor the content of HBV DNA, HbsAg, and HBeAg at 2, 4, 6, 8 and 10 days post-transfection. The effects on cell metabolism were detected by four methyl thiazolyl tetrazolium assay. The locked nucleic acid had an obvious effect on HBV DNA replication and HBsAg and HBeAg expression in a dose and time dependent man-ner. The inhibition rates were 52.14, 57.48, and 29.63% after 6 days, respectively. The locked nucleic acid had no significant effect on cell metabolism. The HBV S gene polypurine region locked nucleic acid could effectively inhibit the replication of HBV in vitro, and could provide an effective target for the treatment of HBV and a theoretical and experimental basis for anti-gene therapy.

The co-transplantation of human bone marrow stromal cells and embryo olfactory ensheathing cells as a new approach to treat spinal cord injury in a rat model.

BACKGROUND: Both bone marrow stromal cells (BMSC) and olfactory ensheathing cells (OEC) have been demonstrated experimentally as promising for therapy of spinal cord injury (SCI). However, clinical use may be constrained by the margin neuronal differentiation capacity of BMSC as well as the limited number of isolatable OEC. This study therefore tested the efficacy of co-grafting human BMSC and OEC in treating thoracic SCI. METHODS: Rat SCI models were created with cushion forces. OEC were labeled with Hoechst 33342 and BMSC with BrdU or GFP. BMSC, OEC and BMSC plus OEC were injected into the injured sites of rat spinal cords. Histologic, electrophysiologic and functional approaches were applied to assess the effects of transplantation of these cell types. RESULTS: Behavioral evaluation showed an improvement in animals with all cell-based treatments. The co-graft led to significantly higher gait scaling. The latency of transcranial magnetic motor-evoked potential (tcMMEP) responses was also better restored in the co-graft group. Larger numbers and sizes of axon bundles through the transitional zone between the normal and injured regions were observed in the co-graft animals in comparison with all other animals. Transplanted bone marrow stromal cells were identified as neurofilament-positive in the co-grafted animals although the number of glial fibrillary acidic protein-positive cells remained the same in all groups. DISCUSSION: Taken together, our results suggest that the combined use of BMSC and OEC may provide an improved approach for the treatment of SCI.

Bone marrow-derived mesenchymal stem cells decrease acute graft-versus-host disease after allogeneic hematopoietic stem cells transplantation.

Graft-versus-host disease is a major complication of allogeneic hematopoietic stem cells transplantation, leading to serious morbidity and mortality. Mesenchymal stem cells(MSC)from bone marrow cause immunoregulation in vitro and in vivo. They also have the potential to protect from lethal GVHD after both autologous and allogeneic Hematopoietic Stem Cells Transplantation (HSCT). In this study, we investigated the mechanisms responsible for GVHD in the allo-HSCT co-transplantation with MSC condition. The model of acute GVHD in Rats was established using allogeneic HSC with donor-derived T cells transplantation, with or without additional donor-derived MSC co-transplantation. The degrees of GVHD were compared, the differentiation of CD4+, CD8+, Th1/Th2 and CD4+CD25+ T cells in vivo were assessed by flow cytometry and RT-PCR analyses. We found that MSC inhibited lethal GVHD after allo-HSCT. The value of CD8+ and CD4+ T cells and the ratio of Th1/Th2 T cell subsets decreased, at the same time the proportion of CD4+CD25+ T cells increased both in spleen lymphocytes and thymocytes in vivo after allo-HSCT with MSC co-transplantation compared with conventional allo-HSCT. Our results strongly suggested that BM-derived MSC has the function of preventing lethal GVHD after allo-HSCT by means of homeostasis of T subsets in vivo.

Implantation of BM mesenchymal stem cells into injured spinal cord elicits de novo neurogenesis and functional recovery: evidence from a study in rhesus monkeys.

BACKGROUND: Transplantation of mesenchymal stem cells (MSC) in rodent models has proved to be an effective therapeutic approach for spinal cord injury (SCI). However, further studies in primate models are still needed before clinical application of MSC to patients. METHODS: MSC were isolated from rhesus monkey BM and induced ex vivo to differentiate into neural lineage cells. Induced cells were labeled with Hoechst 33342 and injected into the injured sites of rhesus SCI models. Function of the injured spinal cord was assessed using Tarlov behavior assessment, sensory responses and electrophysiologic tests of cortical somatosensory-evoked potential (CSEP) and motor-evoked potential (MEP). In vivo differentiation of the implanted cells was demonstrated by the presence of neural cell markers in Hoechst 33342-labeled cells. The re-establishment of the axonal pathway was demonstrated using a true blue (TB) chloride retrograde tracing study. RESULTS: Monkeys achieved Tarlov grades 2-3 and nearly normal sensory responses 3 months after cell transplantation. Both CSEP and MEP showed recovery features. The presence of the neural cell markers neurofilament (NF), neuro-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) was observed in approximately 10% of Hoechst 33342-labeled cells. TB, originally injected at the caudal side of injured sites, was traceable in the rostral thoracic spinal cord, red nucleus and sensory motor cortex. DISCUSSION: Our results suggest that the implantation of MSC-derived cells elicits de novo neurogenesis and functional recovery in a non-human primate SCI model and should harness the clinical application of BM MSC in SCI patients.

Noninvasive detection of endothelial dysfunction in elderly with NIDDM by ultrasonography.

We compared endothelial function noninvasively in 15 elderly patients with noninsulin-dependent diabetes mellitus (NIDDM) with 12 nondiabetic elderly patients. Using high resolution ultrasound, we measured flow-mediated dilatation (FMD) of the brachial artery and sublingual nitroglycerin (NTG)-mediated dilatation. FMD was significantly impaired in subjects with NIDDM compared with control subjects. In NIDDM subjects, FMD with complications was lower than in subjects with noncomplications. However, there was no significant difference in NTG between NIDDM subjects and the control group. Hyperemic blood flow change was lower in NIDDM subjects, but not significant. These results suggest that elderly patients with NIDDM have impaired L-arginine/nitric oxide (NO) pathways and the possibility of capillary rarefaction.

A new noninvasive method for evaluation of coronary endothelial function in hypertensive patients based on change in diameter of the left main coronary artery induced by cold pressor test using echocardiography.

BACKGROUND: Coronary endothelial function is frequently studied by measuring the vasodilator response of coronary arteries to acetylcholine or to cold pressor test by invasive quantitative coronary angiography. Because invasive methods have substantial inherent limitations, studies should attempt to evaluate coronary endothelial function noninvasively. HYPOTHESIS: We attempted to evaluate the accuracy of measurement of the percent change in diameter of the left main trunk induced by cold pressor test with two-dimensional (2-D) echocardiography. Furthermore, we applied this method to the evaluation of coronary artery endothelial function in hypertensive patients. METHODS: We measured the left main trunk diameter in 21 subjects (51 +/- 4 years) before and after cold pressor test using quantitative coronary angiography followed immediately by 2-D echocardiography. The accuracy of measurement of the left main trunk diameter and its percent change by echocardiography was evaluated by comparing the values obtained by the two methods. In addition, using echocardiography, we compared left main trunk diameter responses to cold pressor test in 16 hypertensive patients [51 +/- 5 years (mean +/- standard deviation)] and 16 matched healthy subjects (50 +/- 4 years). RESULTS: Although there was only a weak correlation between the absolute values of the left main trunk diameter measured by the two methods (r = 0.61; p = 0.04), a strong correlation was found between the percent change in diameter measured by the two methods (r = 0.93; p = 0.0001). The percent change in diameter of the left main trunk induced by cold pressor test in hypertensive patients (-3.7 +/- 10.6%) was significantly lower than that in control subjects (13.2 +/- 6.8%, p = 0.0001). CONCLUSIONS: Percent change in diameter of the left main trunk induced by cold pressor test can be evaluated accurately using 2-D echocardiography. Our study showed reduced vasodilation or vasoconstriction of the left main trunk after cold pressor test in hypertensive patients compared with healthy subjects, indicating impaired coronary endothelial function in hypertensive patients. The present echocardiographic method is a potentially useful new noninvasive method for evaluating coronary endothelial function.

Evaluation of endothelial function in hypertensive elderly patients by high-resolution ultrasonography.

BACKGROUND: Multiple investigations, both in experimental models and in middle-aged patients with essential hypertension, demonstrate impaired endothelium-dependent vasodilatation. HYPOTHESIS: We attempted to determine whether hypertension still exerts additional negative effect on endothelial function of large arteries in hypertensive elderly patients who may already be affected by endothelial dysfunction due to aging. METHODS: We compared 13 elderly patients with hypertension [69 +/- 9 years, (mean +/- standard deviation)] with 13 matched healthy elderly subjects (72 +/- 6 years) as controls. Using high-resolution vascular ultrasound, we measured brachial artery responses to reactive hyperemia (with increased flow causing endothelium-dependent dilatation) and sublingual nitroglycerin (causing endothelium-independent dilatation). RESULTS: Flow-mediated dilatation correlated inversely with age (r = -0.60, p = 0.03) in the controls. Flow-mediated dilatation was significantly impaired in hypertensive elderly patients (6.7 +/- 3.3 vs. 13.3 +/- 1.8% in controls, p < 0.0001). No significant difference could found in nitroglycerin-induced dilatation between controls (12.1 +/- 4.9%) and hypertensive elderly patients (10.2 +/- 6.8%, p = 0.5). On multivariate analysis, flow-mediated dilatation in hypertensive elderly patients was inversely related to aging (r = -0.37, p = 0.04) and mean blood pressure (r = -0.57, p = 0.03). CONCLUSIONS: Our study showed decreased flow-mediated dilatation with aging even in the healthy controls, and further decline in flow-mediated dilatation in hypertensive elderly patients compared with controls. This impairment of flow-mediated dilatation in hypertensive elderly patients was related to age and mean blood pressure, indicating that aging and hypertension may independently impair endothelial function in the brachial artery of these patients.

[Effect of jishentang on renal lesions in diabetic rats].

OBJECTIVE: To observe the ameliorative effect of Jishentang (JST) on renal lesions in experimental diabetic rats. METHODS: The diabetic rats were randomly divided into three groups: group with out treatment, group with enalapril treatment (the dose of enalapril was 1 mg/kg.d) and group with JST treatment (the dose of JST was 22.4 g/kg.d). The treatment lasted for 8 weeks. RESULTS: (1) JST treatment attenuated the renal hypertrophy and ratio of kidney and body weight (P < 0.01). (2) JST treatment reduced the levels of SCr and BUN and the levels of Alb, beta 2-m and the activity of NAG in urine in diabetic rats significantly (P < 0.01). The levels of TG and TCh decreased and the levels of HDL-C increased in the serum of the JST treated diabetic rats (P < 0.01). (3) The glomerular basement membrane thickening and volume density of PAS positive staining in mesangial area were significantly decreased in JST treated diabetic rats. CONCLUSIONS: JST exerted obvious ameliorative effect on renal function and structure in diabetic rats.

Determination of mitral valve area in patients with mitral stenosis by the flow-convergence-region method during changing hemodynamic conditions.

Twenty-eight patients with mitral stenosis underwent Doppler echocardiography at rest and during exercise to determine the accuracy of mitral valve area determination by the flow-convergence-region method during exercise-induced changing hemodynamic conditions. The mitral valve area calculated by using the flow-convergence-region method correlated strongly with that measured by the Gorlin formula both at rest (r = 0.85) and during exercise (r = 0.92) for all 28 patients studied. Although mitral valve area obtained by the flow-convergence-region method did not change (p = 0.1) in 16 patients with echocardiographic mitral scores > or = 12, it increased significantly during exercise (p = 0.0001) in 12 patients with echocardiographic mitral scores < 12. This study suggests that in mitral stenosis, the mitral valve area can be accurately estimated by the flow-convergence-region method both at rest and during changing hemodynamic conditions induced by supine bicycle exercise.

Four-dimensional echocardiography: methods and clinical application.

4DE (i.e., dynamic three-dimensional) echocardiography is a new developing technique in recent years. In our study, a three-dimensional echo scan computer system was used to acquire and store the two-dimensional information, then to reconstruct the stereoscopic image of the heart according to its space-time continuum. It can yield a better approach, which can help identify the various structures of the heart and great arteries and facilitate understanding of spatial relations and motion. In addition, it can display physiologic information such as the direction, course, size, and shape of the blood flow. We have examined 138 patients by both transthoracic and transesophageal approaches. Our preliminary experience shows that 4DE is of great value in diagnosing congenital heart disease and valvular disease.

Application of the flow convergence region method to the determination of stroke volume and cardiac output in patients with mitral stenosis.

OBJECTIVE: This study was undertaken to investigate the feasibility and accuracy of determination of stroke volume and cardiac output by calculating transmitral flow volume using the flow convergence region method in patients with mitral stenosis. PATIENTS AND INTERVENTIONS: Fifty-six patients with rheumatic mitral stenosis were studied using imaging and Doppler echocardiography. Aliasing velocities of 20 to 23 cm/s were used to record the flow vonvergence region proximal to the stenotic mitral orifice. The stroke volume (mL) was calculated by multiplying peak transmitral flow rate which was obtained using an angle-corrected hemispheric flow convergence equation, by transmitral velocity time integral (cm) divided by peak transmitral velocity (cm/s) recorded using continuous wave Doppler method. MAIN RESULTS: Stroke volume calculated using the flow convergence region method was not significantly different from that calculated using aortic Doppler two-dimensional echocardiographic method in 39 patients with pure mitral stenosis (75+/-19 [mean+/-1SD] versus 73+/-19 mL, P=0.12), and from that calculated using pulmonic Doppler two-dimensional echocardiographic method in nine patients with mitral stenosis with associated>2+ aortic regurgitation (77+/-12 versus 75+/-14 mL, P=0.49). No significant difference existed between the cardiac output obtained using the flow convergence region method and that obtained using Fick method in 12 patients with pure mitral stenosis. The stroke volume was overestimated by the flow convergence region method when compared with those obtained using aortic Doppler two-dimensional echocardiographic method in patients with mitral stenosis with associated >2+ mitral regurgitation (123+/-40 versus 67+/-15 mL, P=0.001). CONCLUSIONS: The present study provided an alternative way to calculate the stroke volume and cardiac output in patients with mitral stenosis.

Left coronary artery fistula identified by color Doppler flow imaging.

Coronary artery fistula (CAF) is an abnormal communication between a coronary artery and a cardiac chamber, great vessel, or other vascular structures. The presence of the fistula is usually identified by angiography. In this paper, the diagnosis of left coronary artery-right ventricle fistula was made by color Doppler before angiography? and it was confirmed by surgery. Color Doppler flow imaging is a non-invasive method which can reveal the proximal dilatation, the course and the draining site of CAF.

Estimation of mitral valve area in patients with mitral stenosis by the flow convergence region method: selection of aliasing velocity.

OBJECTIVES: We attempted to determine the most suitable aliasing velocity for applying the hemispheric flow convergence equation to calculate the mitral valve area in mitral stenosis using a continuity equation. BACKGROUND: The flow convergence region method has been used for calculating mitral valve area in patients with mitral stenosis. However, the effect of varying aliasing velocity on the accuracy of this method has not been investigated fully. METHODS: We studied 42 patients with mitral stenosis using imaging and Doppler echocardiography. Aliasing velocities of 17, 21, 28, 34, 40 and 45 cm/s were used. The transmitral maximal flow rate (Q [ml/s]) was calculated using the hemispheric flow convergence equation Q = 2 x pi x R2 x AV x alpha/180, where R (cm) is the maximal radius of the flow convergence region, AV is the aliasing velocity, and alpha/180 is a factor accounting for the inflow angle (alpha). Mitral valve area (A [cm2]) was calculated according to the continuity equation A = Q/V, where V (cm/s) is the peak transmitral velocity by the continuous wave Doppler method. RESULTS: Mitral valve area was progressively underestimated with increasing aliasing velocity. The actual and percent differences noted between the mitral valve area by the flow convergence region method and that by two-dimensional echocardiographic planimetry were -0.06 +/- 0.23 cm2 (mean +/- SD) and 0.09 +/- 15.7% at an aliasing velocity of 21 cm/s, increasing gradually with increasing aliasing velocity, and were -1.24 +/- 0.9 cm2 and -72.56 +/- 16.4% at an aliasing velocity of 45 cm/s. Mitral valve areas estimated by the flow convergence region method at an aliasing velocity of 21 cm/s in 11 patients with associated > 2+ mitral regurgitation (2.12 +/- 1.17 cm2) and 8 with associated > 2+ aortic regurgitation (1.28 +/- 0.71 cm2) were not significantly different using planimetry (2.24 +/- 1.39 cm2, p > 0.05 and 1.27 +/- 0.74 cm2, p > 0.05, respectively) but were significantly different by the pressure half-time method (1.59 +/- 1.12 cm2, p < 0.001 and 1.63 +/- 0.93 cm2, p < 0.01, respectively). CONCLUSIONS: This study indicated the most appropriate aliasing velocity for the accurate estimation of mitral valve area in patients with mitral stenosis.

Estimation of regurgitant flow volume based on centerline velocity/distance profiles using digital color M-Q Doppler: application to orifices of different shapes.

OBJECTIVES: In this study we investigated the centerline velocity profile method for flow computation as applied to noncircular, as well as circular, orifices using digital color flow data. BACKGROUND: Recently it has been suggested that flow volume through an orifice can be estimated more accurately by computing the axial "centerline" flow velocity/distance profile proximal to the orifice. METHODS: A total of seven different orifices were mounted in a constant-flow model: four circular orifices, two rectangular orifices with a major/minor axis ratio of 4:1 and 8:1 and an ovoid orifice having a major/minor axis ratio of 2:1. Three different flow rates were examined (1.68, 3.48 and 6.48 liters/min). Digital measurements of flow velocity at discrete positions along the centerline progressing toward the orifice were analyzed to yield complete flow velocity profiles for each orifice at each flow rate. RESULTS: A clear separation of the flow profiles for the three different flow rates was observed independent of orifice size for all of the circular orifices. The velocity/distance acceleration curves showed highly significant correlations using multiplicative regression fits (y = ax-b, r = 0.94 to 0.99, all p < 0.0001). An equation for quantitatively correlating the a and b coefficients from the multiplicative regression fits with flow rates was derived from stepwise regression analysis: Flow rate = 23a + 3.3b - 1.5 (r = 0.97, p < 0.0001, SEE 0.46 liter/min). CONCLUSIONS: In view of the various sizes and shapes encountered clinically for regurgitant orifices, the simplicity of this method for the estimation of the severity of regurgitant lesions might be of importance for clinical applications of this method.

Clinical application of three-dimensional transesophageal echocardiography.

Three-dimensional transesophageal echocardiography is a new and evolving cardiac imaging technique. We reported our experiences of its clinical applications in 59 patients. A series of special temporal longitudinal views were selected by the frame grabber. Then the computer connected each digitized endocardial surface of the longitudinal views according to their spatial position and reconstructed the three-dimensional, cardiac shaded picture with gray scale. The three-dimensional transesophageal echocardiographic images were divided into three areas. The right area was right anterior to the esophagus and included such structures as the superior vena cava, right atrium, interatrial septum, and left atrium; the size, shape, and location of an atrial septal defect could be clearly shown. In the middle area the origin and the course of the two great arteries could be visualized, thus facilitating the diagnosis of transposition of the great arteries; in patients with obstruction of the right ventricular outflow tract, the circular ridgelike narrowing in the right ventricle was clearly visualized. In the left area the contour and size of the left ventricle and left atrium and the shape and point of coaptation of the mitral valve could be demonstrated; in patients with mitral valve prolapse, part of either leaflet protruded into the left atrium and appeared as a spoonlike depression in the mitral valve. Other entities subjected to three-dimensional transesophageal echocardiographic reconstruction included cor triatriatum, left atrial myxoma, aneurysm of sinus of Valsalva, dissecting aortic aneurysm, mitral stenosis, mitral regurgitation, and mitral valve prolapse.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of ventricular septal defect by transesophageal echocardiography: intraoperative assessment.

The miniaturization of transesophageal echocardiography (TEE) probes, together with the development of the capability for biplane imaging from the esophagus, have increased the use of TEE in pediatric cardiology. The aim of this study was to evaluate the TEE findings in patients with ventricular septal defect (VSD) before and after closure primarily by means of pediatric biplane probes. This study group included 69 patients who underwent VSD closure as an isolated repair or as a part of a definitive repair of a more complicated lesion. Ages ranged from 6 days to 15.6 years (median 1 year, 4 months), with operative weights ranging from 2.9 kg to 68 kg (median 10 kg). Preoperative and follow-up transthoracic echocardiograms (TTE) were also performed. Intraoperative TEE was performed without complication in all 69 patients. Preoperative results: (1) anatomic findings: Two muscular VSDs were detected by matrix TEE but could not be observed by TTE. A patient with preoperative TTE diagnosis of an ostium primum ASD was found to have atrioventricular (AV) canal by TEE. In three of six AV canal type VSDs, both TTE and TEE demonstrated left ventricular-right atrial shunting (2) aortic regurgitation associated with VSD: Aortic regurgitation as a result of right coronary cusp prolapse was detected in one of five supracristal VSDs in which the biplane or matrix TEE was used.(ABSTRACT TRUNCATED AT 250 WORDS)