超声组织定征在心血管疾病诊断中的应用

超声组织定征(Ultrasonic Tissue Characterization,UTC)技术是通过检测组织的声学参数来定量描述正常和病理组织的物理(声学)特性。研究表明,背向散射积分作为组织定征的参数,可以识别缺血心肌、顿抑心肌、梗死心肌、左心室心肌肥厚及心脏移植排斥反应,评价和分析系统性疾病如糖尿病等引起的弥漫性心肌受累的状态、心腔内血栓和动脉斑块的成份等。因而UTC技术具有很大的临床价值和发展潜力,将成为常规超声心动图诊断的辅助手段。本文就超声组织定征技术在心血管疾病诊断中的应用及进展作一综述。     

Ultrasonic tissue characterization of normal and ischemic myocardium

Cardiac ultrasonic tissue characterization is designed to use the alterations in acoustic signals from the myocardium to differentiate normal from ischemic or infarcted tissue due to their characteristic backscatter attenuation. Various approaches such as use of a gray scale, color display, or quantitative image analysis have been used for tissue characterization, but all depend on subjective assessments and are not necessarily reproducible. The most promising method has been the use of "raw" radiofrequency signals and measure changes in the ultrasonic attenuation with an index of backscatter to distinguish normal from abnormal myocardium called "integrated backscatter" (IB). Various studies have demonstrated the changes in the ultrasonic backscatter with ischemia or infarction. In this review we summarize our experience with a research prototype instrument in tissue characterization and differentiation of normal, ischemic, infarcted, and post ischemic reperfused myocardium in anesthetized open chest dogs. Currently we are investigating the role of ultrasonic tissue characterization to estimate infarct size and plan to apply these observations to patients in order to detect viable myocardium and quantitate infarct size.     

Tissue characterization with intra-arterial ultrasound: Special promise and problems

Although we are able to identify many tissue types based on the screen image in intravascular ultrasound, there is additional information in the ultrasound signal which could be of assistance in characterization and identification of tissue. Intravascular ultrasound has several special characteristics which affect tissue characterization. These include the high transducer frequency, small transducers, short and relatively uniform path to the tissue, and limited tissue types to identify. These characteristics influence the results obtained by absolute backscatter, local statistics, frequency dependent backscatter, and angle dependency of backscatter. These effects are both positive and negative, and in many cases can be observed in clinical imaging. Another area of tissue characterization which can be performed with ultrasound is measurement of arterial wall elasticity. This can be of importance in the evaluation of mechanisms of dilatation, and the potential for complications.     

Tissue Characterization by Ultrasound: What is Possible Now? What Will be Possible?

The purpose of this review is to discuss the principles of ultrasound tissue characterization. We describe gray scale analysis, backscatter techniques, real-time backscatter imaging, enhancement of abnormal tissue properties with contrast agents or liposomes, and use of acoustic microscopy. Ultrasound tissue characterization offers the promise of direct identification of abnormalities of the myocardium without relying on indirect manifestations such as abnormalities in cardiac function.     

Myocardial ultrasonic tissue characterization for estimating histological abnormalities in hypertrophic cardiomyopathy: comparison with endomyocardial biopsy findings.

AIMS: In this study, we investigated the clinical usefulness of ultrasonic tissue characterization with integrated backscatter for the evaluation of myocardial histological abnormalities in comparison with endomyocardial biopsy findings in patients with hypertrophic cardiomyopathy. METHODS: Twenty patients with hypertrophic cardiomyopathy and 20 normal subjects were enrolled in this study. We measured two parameters for the ultrasonic tissue characterization with integrated backscatter: the magnitude of the cardiac-cycle-dependent variation in integrated backscatter signals (cdv-IB) and the mean value of integrated backscatter signals calibrated by the pericardium (cal-IB). These parameters were measured at both the interventricular septum and the left ventricular posterior wall. Histological findings of right ventricular endomyocardial biopsy specimens were analyzed by computer image analyzer. RESULTS: cdv-IB was significantly lower and cal-IB significantly higher in both the interventricular septum and the left ventricular posterior wall in patients with hypertrophic cardiomyopathy compared with normal subjects. In patients with hypertrophic cardiomyopathy, the degree of myocardial disarray, interstitial fibrosis, and nonhomogeneity of myocyte size showed positive correlations with cal-IB and negative correlations with cdv-IB. CONCLUSIONS: Ultrasonic tissue characterization with IB enables the noninvasive evaluation of myocardial histological abnormalities in patients with hypertrophic cardiomyopathy.     

Integrated backscatter during harmonic and fundamental frequency imaging--effect of depth,mechanical index,and tissue anisotropy: implications for myocardial tissue characterization

OBJECTIVE: To explore the potential advantages of tissue harmonic imaging (THI) versus fundamental frequency imaging (FFI) when applied to tissue characterization. METHODS: A Philips Medical Systems Sonos 5500 echocardiograph equipped with a broadband transducer (S4) and an on-line quantitative analysis software package (Acoustic Densitometry) was used for imaging. The effect of mechanical index (MI), imaging depth, and anisotropy on relative backscatter amplitude was evaluated. RESULTS: This study demonstrated that imaging with tissue harmonics generated relatively greater backscatter values at clinically relevant imaging depths and instrument settings referenced to FFI. This effect was dependent on MI setting. A direct relationship between backscatter amplitude and MI was demonstrated. Additionally, tissue anisotropy had similar effects on integrated backscatter amplitude during both THI and FFI. However, relative backscatter values at each fiber orientation are greater during THI at similar instrument settings when referenced to FFI. CONCLUSION: Tissue harmonic imaging may offer advantages over FFI for myocardial tissue characterization.     

老年高血压心肌背向散射成像声学密度定量的临床应用

目的　应用声学密度定量 (AD)方法对老年高血压病人心肌超声背向散射成像 (IBS)进行分析 ,探讨其临床诊断应用价值及意义。方法　按 1 999年 WHO高血压治疗标准并应用超声心动图仪检测 64例老年男性住院病人 ,其中正常组 31例 ,高血压组 33例 (高血压病史均 >3年且治疗时间均 >1年 ) ,比较各组室间隔心肌 IBS的 AD诸参数。结果　 Loop和两种触发方式声学密度曲线的峰值密度 (pi)、曲线下面积 (auc)高血压组均高于正常组 (均 P<0 .0 5)。结论　长期高血压和衰老致心肌组织纤维化进一步加重 ,致声学密度参数显著改变 ,其可作为简单、有效且无创的研究心肌病理学方面改变的辅助检测方法     

Quantitative Texture Analysis in Two-Dimensional Echocardiography

Background: Myocardial reflectivity is abnormally increased in patients with thalassemia major under transfusion treatment, probably due to myocardial iron deposits and / or secondary structural changes. Such increased reflectivity has been detected by both qualitative and subjective analysis of two-dimensional echocardiographic (2-D echo) images and quantitative assessment of integrated backscatter amplitude with noncommercially available ultrasound prototypes. The purpose of this study was to assess the acoustic properties of myocardium in patients with beta-thalassemia major and iron overload by means of quantitative computerized offline textural analysis of conventionally recorded 2-D echo images, and to compare textural data with other qualitative (visual assessment) and quantitative (ultrasound backscatter analysis) approaches for myocardial ultrasound tissue characterization simultaneously applied to these patients. Methods and Results: Thirty-five young patients with thalassemia major, without clinical signs of cardiac failure, and 20 age and sex matched normal controls were studied by echocardiography. Each patient was receiving blood transfusion every 2-3 weeks. Two-dimensional echo images, obtained with a commercially available echocardiograph using the parasternal long-axis view, were digitized offline and analyzed by first and second order texture algorithms applied to regions of interest in the myocardium (septal and posterior wall). The mean gray level value was higher in thalassemic patients than in controls on both the septum (110 ± 25 vs 57 ± 13, arbitrary units on a 0-255 scale; P > 0.01) and posterior wall (91 ± 25 vs 67 ± 18; P > 0.01). Among second order statistical parameters, contrast and angular second moment significantly (P > 0.01) differentiated septal and posterior walls of patients and controls. In thalassemic patients, no consistent correlation was found between wall texture parameters and hematologic (years of transfusions and chelation, number of transfusions), 2-D echo (posterior wall thickness, left ventricular end-diastolic diameter), and Doppler (transmitral E/A waves ratio) parameters. Myocardial walls with visually assessed increased echo reflectivity showed a trend toward higher values of mean gray level when compared with myocardial segments with qualitatively assessed normal reflectivity (septum: 121 ± 26 vs 106 ± 24; posterior wall: 105 ± 23 vs 87 ± 23). Although radiofrequency integrated backscatter has been demonstrated to be capable of identifying thalassemic patients, no significant correlation was found between mean gray level (by texture analysis) and radiofrequency data (septum: r = 0.03; posterior wall: r = 0.09; P = NS for both). Conclusions: Myocardial walls affected by hemochromatosis show ultrasound image texture alterations that may be quantified with digital image analysis techniques and appear mostly unrelated to hematologic and conventional, as well as radiofrequency-based, echocardiographic parameters. These changes in quantitatively evaluated echo reflectivity are present even before the development of clinical and echocardiographic signs of cardiac dysfunction.     

Abnormal Ultrasonic Tissue Characterization of the Myocardium in Children Receiving Doxorubicin for Cancer Therapy

Ultrasonic tissue characterization by integrated backscatter is a sensitive tool to detect myocardial changes related to specific diseases. Cardiotoxicity related to doxorubicin use is well known and remains a major concern. To determine if ultrasonic tissue characterization of the myocardium is abnormal in patients receiving doxorubicin, we studied the myocardium of pediatric patients receiving doxorubicin by a real-time integrated backscatter (IB) imaging system. Three values of IB parameters were averaged from the left ventricular posterior wall at the level of the tip of the mitral valve. In addition to standard echo parameters, we obtained the following IB parameters: peak, nadir, cyclic variation (CV), end-diastole, heart-rate corrected delay of nadir (Delay_c), and the ratio of CV over end-diastole. IB parameters were normalized as Z scores from multiple linear regression equations including echo wall thickness and functional indices from a normal control group of 72 children. We evaluated 27 patients at a median age of 11.6 years (1.6 years to 20.3 years) and median time of 1.7 m (2 days to 7.2 years) after a mean cumulative dose of doxorubicin of 188 ± 120 mg/m² for treatment of neoplasm. Mean (± SD) Z scores for IB variables were as follows: zPeak 0.15 ± 1.07, P = 0.47; zNadir 0.41 ± 1.16, P = 0.08; zCV -0.49 ± 0.95, P = 0.01; zEnd-diastole 0.17 ± 0.94, P = 0.38; zDelay_c 0.33 ± 0.80, P = 0.06, and zCV/Peak -0.59 ± 1.06, P = 0.009. This study shows that ultrasonic IB of the myocardium of children receiving doxorubicin is abnormal and is independent of the cumulative dosage of doxorubicin or the amount of time since the last dose.     

On-Line Myocardial Tissue Characterization with a New Commercially Produced Software

Myocardial tissue characterization has been performed using various ultrasonic techniques, one of which is the cyclic variation of integrated backscatter, a method that analyzes the acoustic properties of the myocardium using backscattered radiofrequency signals to provide information about myocardial structure and function. Previous studies using prototype equipment have demonstrated a reduction in the cardiac cycle variation of integrated backscatter in various pathologic states. Recently, a commercially produced software package that allows online analysis of cyclic variation of integrated backscatter has been made available for testing by various investigators. To evaluate this new commercially produced software, we compared integrated backscatter results in three groups of patients: a control group; an end-stage cardiomyopathy group; and a heart transplant recipient group. Integrated backscatter of the septum and posterior walls in the parasternal long axis and 12, 3, 6, and 9 o'clock regions in the short axis was performed using a commercially produced program (Hewlett-Packard Sonos 1500). In the control group, the mean cyclic variation of integrated backscatter was 5.04 +/- 1.60 dB in the septum and did not significantly vary from the rest of the regions studied. In comparison, cyclic variation of integrated backscatter in every region studied was reduced in the cardiomyopathy and heart transplant groups. Intraobserver variability, interobserver variability, and reproducibility over a 3-month interval was found to be 6.5%, 5.7%, and 7.5%, respectively. These results indicate that: (1) online analysis of cardiac cyclic variation of integrated backscatter is possible utilizing commercially produced software; (2) results obtained are consistent with a low intraobserver and interobserver variability and are reproducible over time; and (3) as observed in the comparison between the transplant and control groups, this information may detect changes in cardiac structure even in the absence of changes in function. (ECHOCARDIOGRAPHY, Volume 13, May 1996)     

Structural remodeling of human myocardial tissue after infarction. Quantification with ultrasonic backscatter.

BACKGROUND. Remodeling of myocardial tissue after infarction may culminate in the development of either a well-healed scar or a thin, expanded heart wall segment that predisposes to ventricular aneurysm formation, congestive heart failure, or ventricular tachycardia. The three-dimensional architecture of mature human infarct tissue and the mechanisms that determine it have not been elucidated. We have previously shown that quantitative ultrasonic backscatter can be used to define the transmural organization of human myofibers in the normal ventricular wall by measuring the dependence of backscatter on the angle of insonification, or ultrasonic anisotropy. We propose that measurement of ultrasonic anisotropy of backscatter may permit quantitative characterization of the transmural architecture of tissue from areas of myocardial infarction and facilitate identification of fundamental mechanisms of remodeling of the ventricular wall. METHODS AND RESULTS. We measured integrated backscatter in 33 transmural sections from 12 cylindrical biopsy specimens (1.4-cm diameter) sampled from central regions of mature infarction in six explanted fixed human hearts. Tissue samples were insonified in two-degree steps around their entire circumference at successive transmural levels with a 5-MHz broad-band piezoelectric transducer. Backscatter radio frequency data were gated from the center of each specimen, and spectral analysis was performed on the gated radio frequency for the computation of integrated backscatter. Histological morphometric analysis was performed on each specimen for determination of the predominant fiber orientation and the percentage of tissue infarcted at consecutive transmural levels. The average percentage of tissue infarcted for all transmural levels was 49 +/- 3% (range, 13-80%). Histological attributes varied from patchy fibrosis to extensive confluent zones of scar tissue. The angle-averaged integrated backscatter for all transmural levels in infarct tissue was approximately 5 dB greater than that previously measured in normal tissue in our laboratory (-48.3 +/- 0.5 versus -53.4 +/- 0.4 dB, infarct versus normal). Marked anisotropy of backscatter was observed in tissue from areas of infarction and was characterized by a sinusoid-like dependence on the angle of insonification at each transmural level. Insonification perpendicular to infarct fibers yielded values for integrated backscatter 14.8 +/- 0.5 dB greater than those for insonification parallel to these fibers. Juxtaposition of the sinusoid-like anisotropy functions from all consecutive transmural levels demonstrated a progressive shift in the orientation of scar tissue elements from epicardial to endocardial levels of 14.6 +/- 1.5 degrees/mm of tissue. The transmural shift in fiber orientation per millimeter of tissue from the area of infarction exceeded that previously measured for normal tissue (9.2 +/- 0.7 degrees/mm) by 59%. This marked augmentation in angular shift per millimeter of tissue results from a generalized structural rearrangement (or reorientation) of fibers across the entire ventricular wall in the infarct zone that we hypothesize is determined in part by dynamic mechanical forces, imposed by the surrounding functional normal tissue, that tether the "infarcted" tissue. CONCLUSIONS. Myocardial tissue from areas of myocardial infarction manifests substantial anisotropy of ultrasonic scattering that may be useful for quantitative characterization of the alignment and overall three-dimensional anatomic organization of mature infarct scars.     

Quantitative ultrasonic assessment of normal and ischaemic myocardium with an acoustic microscope: relationship to integrated backscatter

PURPOSE OF INVESTIGATION--The aim was to study ultrasonic propagation properties of normal and ischaemic myocardium with a scanning laser acoustic microscope and to correlate these changes with ultrasonic backscatter. DESIGN--Myocardial ischaemia was produced by total occlusion of left anterior descending coronary artery in anaesthetised open chest dogs. Myocardium supplied by left circumflex coronary artery served as normal control. IBR5, an optimum weighted frequency average (4-6.8 MHz) of the squared envelope of diffraction corrected backscatter, was measured in vivo. Ultrasonic attenuation coefficient, an index of loss per unit distance, the propagation speed and heterogeneity index were measured from normal and ischaemic regions with a scanning laser acoustic microscope which operates at 100MHz in vitro. Myocardial water content of normal and ischaemic myocardium was also estimated. SUBJECTS--Were five anaesthetised mongrel dogs. RESULTS--Attenuation coefficient of 33.8(SD4.2) dB.mm-1 in the ischaemic tissue was lower than 63.8(17.2) dB.mm-1 in the normal tissue (p less than 0.01). Ultrasonic speed was lower in ischaemic than normal myocardium at 1584(25) v 1612(35) m.s-1 (p less than 0.05). Heterogeneity index of 11(7) m.s-1 in the ischaemic region was lower than 14(8) m.s-1 in the normal region (27% reduction, p less than 0.05). IBR5 and myocardial water content were higher in the ischaemic than the normal myocardium: -37.2(SEM1.8) dB v -46.6(0.6) dB, (p less than 0.01) and 80.9(0.0)% v 78(0.2)%, (p less than 0.05) respectively. CONCLUSION--Ultrasonic properties of the myocardium are significantly altered during acute ischaemia.     

Abnormal myocardial acoustic properties in diabetic patients and their correlation with the severity of disease.

Although patients with diabetes mellitus may be afflicted by cardiomyopathy, its prevalence and nature are controversial. Studies have shown that fibrosis alters the acoustic properties of the heart in animals and humans and that the changes are detectable by cardiac tissue characterization with ultrasound. The present study was performed to characterize myocardial acoustic properties in patients with insulin-dependent diabetes to determine whether ultrasound tissue characterization could detect changes potentially indicative of occult cardiomyopathy. The magnitude of cyclic variation of myocardial ultrasound integrated backscatter and its phase delay with respect to the onset of the cardiac cycle in the septum and posterior wall of the left ventricle were measured in 54 patients with diabetes who had no overt cardiac disease. Conventional echocardiography documented normal ventricular systolic function in 96%. As compared with results in age-matched patients without diabetes studied previously, cyclic variation of integrated backscatter was reduced (4.6 +/- 0.8 vs. 3.6 +/- 1.4 dB; p less than 0.001). In addition, delay was significantly increased (0.86 +/- 0.09 vs. 0.99 +/- 0.15). The primary analysis of the data focused on differences among the diabetic patients. Reduction of cyclic variation of backscatter was greatest in patients with diabetes who had neuropathy (3.2 +/- 1.0 dB; p less than 0.001) as was the increase in delay (1.04 +/- 0.16, p less than 0.001 vs. values in patients without neuropathy). Retinopathy and nephropathy were associated with abnormal myocardial acoustic properties as well. Thus, abnormalities that may reflect fibrosis or other occult cardiomyopathic changes in diabetic patients without overt heart disease are readily detectable by myocardial tissue characterization with ultrasound and parallel the severity of noncardiac diabetic complications.     

Hepatocyte volume as an indicator of hepatic functional reserve in cirrhotic patients with liver tumours

Using computed tomography (CT), measurements of whole liver volume have been used for the assessment of pre-operative functional reserve in cirrhotics. However, measurements of hepatocyte volume, which exclude stromal fibrous tissue, are considered to more directly reflect hepatic functional reserve. We investigated the relationship between total hepatocyte volume and each of the parameters of conventional liver function. Indocyanine green (ICG) tests and blood analyses for the assessment of liver function were performed prior to surgery in cirrhotic patients with liver tumours. Pre-operative liver volume was determined by integrating images of each liver area obtained by CT. Liver area was measured by an image processing program that traced the profile of the liver image while excluding the tumorous area. Sections of normal tissue stained by the haematoxylin-eosin method, were obtained from the resected liver. Using these sections, a hepatocyte area: whole tissue area ratio was calculated using the image processing program, by tracing the profiles of the hepatocyte nodules. The total volume of hepatocytes was then calculated by multiplying the liver volume by this ratio. The hepatocyte volume per unit bodyweight was significantly correlated with ICG tests and with many other parameters of normal liver function. However, the liver volume per unit bodyweight was correlated only with the plasma ICG disappearance rate and with the blood platelet count. These observations suggest that the functional reserve of the cirrhotic liver is assessed more precisely by hepatocyte volume than by liver volume.     

Advances in ultrasound methods for high-resolution imaging of the cardiovascular system

Acoustic microscopy entails the use of high-frequency high-resolution ultrasound methods to produce images of sound waves reflected from or propagated through some tissue of interest. The image contrast depends on microscopic differences in the intrinsic material properties of the substance imaged, such as mass density or compressibility. Pathologic changes in cardiovascular tissues at the subcellular level can be observed with high-frequency acoustic imaging techniques, based on alterations in the structure, properties, and organization of cells and their surrounding matrix. Potential applications extend from delineation of cardiovascular development in experimental animals to clinical characterization of the composition of atherosclerotic lesions with intravascular ultrasound and estimation of the potential for plaque rupture and infarction. (Trends Cardiovasc Med 1997;7:168-174). ? 1997, Elsevier Science Inc.     

Feasibility of optical coherence tomography for high-resolution imaging of human gastrointestinal tract malignancies

Optical coherence tomography (OCT) is a new imaging technology which can perform high-resolution, cross-sectional imaging of the internal microstructure of biological tissues. OCT is analogous to ultrasound, except that it measures the intensity of back-reflected infrared light rather than sound waves. OCT performs two- and three-dimensional imaging of tissue microstructure in situ and in real time. It can achieve image resolutions approaching the cellular level over approximately the same imaging depths as a conventional biopsy. In this article we examine the feasibility of OCT for high-resolution imaging of gastrointestinal malignancies with ex-vivo imaging of normal and pathologic microstructures. Tissue, both normal and neoplastic, was obtained from patients undergoing surgical resection after an initial diagnosis of a gastrointestinal malignancy. The tissue samples were imaged prior to fixation using a laboratory OCT system. The OCT system consists of a fiber optic-based Michelson interferometer, a commercially available amplified superluminscent light source, and a computer for data acquisition. The images were subsequently compared with histological cross-sections corresponding to the imaged areas. The stratified squamous epithelium of the normal esophagus was clearly visible in the OCT images and contrasted to the disorganized and non-uniform nature of the mucosal layers of Barrett's esophagus and squamous carcinoma. The columnar epithelial morphology as well as other mucosal structures in normal colon were distinctly visible using OCT. In contrast, disorganization of the normal mucosal layers and ulcerative lesions were identified in tissues from ulcerative colitis and adenocarcinoma of the colon. The ability of OCT to image tissue microstructure at high resolutions makes it a potentially powerful technology for minimally invasive assessment of the gastrointestinal tract and the evaluation of early neoplastic changes. Received: March 25, 1999 / Accepted: July 23, 1999     

Quantitative ultrasonic tissue characterization as a new tool for continuous monitoring of chronic liver remodelling in mice.

BACKGROUND/AIM: Recognition of the limitations of liver biopsies has led to the need for non-invasive tests to assess liver fibrosis from intensity and kinetic point of views. The aim of the present study was to evaluate non-invasive ultrasonic tissue characterization for the continuous monitoring of this process in mice. METHODS: Twelve-week-old male and female C57Bl6/J mice were submitted to repetitive carbon-tetrachloride (CCl4) intraperitoneal injections during 8 weeks or analysed 28 days after common bile duct ligation (BDL). The extent and kinetic of the disease progression were followed by the measurement of ultrasound backscatter intensity. This was compared with histological and blood parameter analysis. RESULTS: CCl4 induced a progressive increase in in vivo liver tissue backscatter intensity in both males and females. This increase was mainly correlated with interstitial fibrosis and, to a lower extent, with nuclear surface of the hepatocytes. A similar result was found after BDL. CONCLUSIONS: These data demonstrate for the first time in a systematic study that ultrasound tissue characterization can be used as a reliable tool to follow liver remodelling in mice continuously.     

Hemodialysis improves myocardial interstitial edema and left ventricular diastolic function in patients with end-stage renal disease: noninvasive assessment by ultrasonic tissue characterization

Congestive heart failure is the most common cause of mortality in patients with end-stage renal disease (ESRD). Ultrasonic tissue characterization with integrated backscatter offers a promising method for the noninvasive assessment of regional myocardial contractile performance and fibrosis. The aim of this study was to investigate the effect of hemodialysis (HD) on myocardial tissue characterization and left ventricular function in ESRD patients. We examined 26 patients with ESRD undergoing routine HD (age 63 ± 12 years, duration of HD 9.2 ± 3.2 years) and 30 patients with essential hypertension (HT; 60 ± 10 years). Routine echocardiographic parameters and the cyclic variation of ultrasonic integrated backscatter of the ventricular septum (CV-IBS) were measured. Left ventricular mass index was significantly larger in patients with ESRD than in those with HT (217 ± 56 vs 146 ± 45 g/m², P < 0.05). The indices for left ventricular diastolic function (E/A, the ratio of left ventricular peak early to late diastolic filling velocity; DT, the deceleration time of the early diastolic filling) and CV-IBS had deteriorated significantly in patients with ESRD before HD compared with those with HT (E/A, 0.6 ± 0.2 vs 0.9 ± 0.3, P < 0.05; DT, 228 ± 23 vs 184 ± 19 ms, P < 0.05; CV-IBS, 9.0 ± 1.3 vs 12.4 ± 0.9 dB, P < 0.05), possibly reflecting interstitial fibrosis. In patients with ESRD, HD reduced calculated left ventricular mass index by 19% (before HD, 217 ± 56 vs immediately after HD, 176 ± 45 g/m², P < 0.05) and CV-IBS by 19% (9.0 ± 1.3 vs 7.3 ± 1.1 dB, P < 0.05), that possibly reflected improvement of interstitial edema. HD also significantly improved indices for left ventricular diastolic function (E/A, 0.6 ± 0.2 vs 0.9 ± 0.2, P < 0.05; DT, 228 ± 23 vs 188 ± 21 ms, P < 0.05). HD improves myocardial interstitial edema and left ventricular diastolic function in patients with ESRD. Noninvasive assessment of ultrasonic tissue characterization is useful in defining the pathophysiological changes of ventricular myocardium in patients with ESRD. Received: December 17, 2001 / Accepted: April 19, 2002 Correspondence to O. Hirono     

Expression of growth hormone receptor and its mRNA in hepatic cirrhosis

AIM: To investigate the expression of growth hormone receptor (GHR) and mRNA of GHR in cirrhotic livers of rats with the intension to find the basis for application of recombinant human growth hormone (rhGH) to patients with liver cirrhosis.METHODS: Hepatic cirrhosis was induced in SpragueDawley rats by administration of thioacetamide intraperitoneally for 9-12 weeks. Collagenase Ⅳ was perfused in situ for isolation of hepatocytes. The expression of GHR and its mRNA in cirrhotic livers was studied with radio-ligand binding assay, RT-PCR and digital image analysis.RESULTS: One class of specific growth hormone-binding site, GHR, was detected in hepatocytes and hepatic tissue of cirrhotic livers. The binding capacity of GHR (RT, fmol/mg protein) in rat cirrhotic liver tissue (30.8±1.9) was significantly lower than that in normal control (74.9±3.9) at the time point of the ninth week after initiation of induction of cirrhosis (n=10, P＜0.05), and it decreased gradually along with the accumulation of collagen in the process of formation and development of liver cirrhosis (P＜0.05). The number of binding sites (×10 4/cell) of GHR on rat cirrhotic hepatocytes (0.86±0.16) was significantly lower than that (1.28±0.24)in control (n= 10, P＜0.05). The binding affinity of GHR among liver tissue, hepatocytes of various groups had no significant difference (P＞0.05). The expression of GHR mRNA (riOD,pixel) in rat cirrhotic hepatic tissues (23.3±3.1) was also significantly lower than that (29.3±3.4) in normal control (n=10, P＜0.05).CONCLUSION: The growth hormone receptor was expressed in a reduced level in liver tissue of cirrhotic rats,and lesser expression of growth hormone receptors was found in a later stage of cirrhosis. The reduced expression of growth hormone receptor was partly due to its decreased expression on cirrhotic hepatocytes and the reduced expression of its mRNA in cirrhotic liver tissue.     

Relationship between hepatocyte proliferation and hepatitis delta virus replication in neoplastic and non-neoplastic liver tissues

SUMMARY. We studied the relationship between hepatocyte proliferation and hepatitis delta virus (HDV) replication at the single cell level. The proliferating cell nuclear antigen (PCNA) (by immunohistochemistry) and the HDV RNA (by in situ hybridization) were stained in neoplastic and non-neoplastic liver tissues of 19 patients with chronic HDV infection, including four cases of cirrhosis with superimposed hepatocellular carcinoma (HCC). As controls, we assessed the hepatocyte proliferation of liver tissues from 16 patients with chronic hepatitis B and on three normal livers. The hepatocyte PCNA labelling index of HDV-infected tissues was comparable with that seen in chronic hepatitis B-infected livers but was significantly higher than that observed in normal livers. Although cirrhotic tissues had lower hepatocyte proliferating fractions than non-cirrhotic tissues, the difference was not statistically significant. The hepatocyte proliferation rate did not correlate with the level of intrahepatic HDV replication or with the histological activity. In double-labelling experiments, PCNA and HDV RNA staining did not co-localize, with the exception of two of three cirrhotic tissues associated with HCC, where the association between the two markers was statistically significant. This co-localization was not observed, however, in the adjacent tumorous tissues. In patients with chronic HDV infection the hepatocyte proliferation was increased with respect to normal liver tissue, but was comparable with that observed in patients with chronic hepatitis B virus infection and did not correlate with the level of HDV replication or the histological activity. In the cirrhotic tissue of patients with HCC (but not in the tumour counterpart), HDV RNA may occasionally co-localize with the marker of hepatocyte proliferation. Whether this association between viral replication and cell division is related to liver carcinogenesis remains to be established.