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Biomedical subjects

X Ouyang

Publications and source records attributed to X Ouyang.

At least 37 records · Page 2Linked to original sources

[Study on the inhibition of nude mice transplantation tumor growth by telomerase ribozyme].

OBJECTIVE: To explore whether telomerase ribozyme could inhibit the growth of nude mice transplantation tumor. METHODS: A xenograft human-nude mouse model was constructed. A total of 24 mice were divided into 4 groups, namely, the saline control(Group 1), the blank plasmid control(Group 2), the 20 microg/mouse/day p(XJ-neo-teloRZ)(Group 3) and the 30 microg/mouse/day p(XJ-neo-teloRZ)(Group 4). All plasmids were packaged by lipofectamine and used for subcutaneous, continuous infusions over 14 days. Tumor measurements and observations of animal behavior were recorded daily. On completion of the study, the mice were killed by cervical dislocation and their organs were removed, weighed, and stored in formaldehyde for histological examination. The telomerase activities of tumor tissues of each group were also detected. RESULTS: The telomerase ribozyme effectively inhibited the telomerase activities of tumor tissues and promoted the apoptosis of tumor cells. The in vivo studies showed a significant decrease in tumor size in mice treated with p(XJ-neo-teloRZ) when compared to the mice treated with controls. Furthermore, the in vivo effect of p(XJ-neo-teloRZ) was dose dependent. CONCLUSION: Telomerase ribozyme is a powerful telomerase inhibition agent; probably it could be effective in tumor gene therapy.

Animals↗

Characterization of the ecdysteroid UDP-glucosyltransferase gene of Spodoptera litura multinucleocapsid nucleopolyhedrovirus.

The ecdysteroid UDP-glucosyltransferase (EGT) gene of Spodoptera litura multinucleocapsid nucleopolyhedrovirus (SpltMNPV), Guangzhouisolate was sequenced. The open reading frame of the gene is 1530 nucleotides long, encoding a putative protein of 509 amino acids with Mr of 58,500. The 5' noncoding region has a TATA box. A polyadenylation signal, AATAAA, is contained downstream of the translation stop codon. A putative signal peptide is present at the N-terminus of the protein. Homology comparison with other baculovirus egt genes showed that these genes are highly conserved. The SpltMNPV egt gene is closely related to that of Spodoptera littoralis nucleopolyhedrovirus (SliNPV) with the identities of 84% in nucleotide and 91% in amino acid, respectively. In this study, an alignment of amino acid sequences has been made and a tentative phylogenetic tree of twelve baculovirus egts is presented.

Amino Acid Sequence↗

[Treatment of class II furcation involvement: guided tissue regeneration with atelocollagen membrane].

OBJECTIVE: To evaluate the effectiveness of guided tissue regeneration (GTR) on class II furcation involvement with absorbable atelocollagen membranes made in China. METHODS: GTR was performed on 26 sites with class II furcation defect in 17 patients. Clinical measurements and digital subtraction radiography were used to evaluate the effect. RESULTS: Clinical measurements showed significant attachment gain, significant decrease of probing depth (PD) and horizontal probing depth (HPD) at 3, 6, and 12 month after surgery. The mean reduction of HPD were 1.8 mm, 2.2 mm, 2.5 mm at 3 month (23 sites), 6 month (12 sites), and 12 month (6 sites) after surgery respectively. PD decreased by 1.22 mm, 1.56 mm and 1.75 mm. Attachment gained by 1.17 mm, 2.11 mm and 1.75 mm. Three sites showed complete defect fill during 3 to 12 months after surgery. Only 1 mm or less could be probed horizontally in another 4 furcation lesions during 3 to 12 months post-operation. The HPD of pre-operation in these 7 cases were 2.0 mm to 4.0 mm, and the bone defects observed during surgery were smaller in these cases than in other cases. The other 19 furcation involvement improved significantly. The result of digital subtraction radiography showed that increase of bone density was found at 2 months after surgery. The amount of bone gain increased with time. The areas of increased bone density were 0.3-25.7 mm2 in 13 teeth. CONCLUSION: The class II furcation involvement can be treated by GTR with absorbable atelocollagen membranes with good results.

Adult↗

Adjacent carboxyterminal tyrosine phosphorylation events identify functionally distinct ErbB2 receptor subsets: implications for molecular diagnostics.

Site-directed mutagenesis can define the effects of altering one or more amino acids within a protein, but this technique may lack sensitivity when used to characterize proteins which differ conformationally or posttranslationally at multiple sites. A novel alternative approach involves the direct characterization of wild-type protein isoforms identified by site-specific immunodetection. To this end we have developed antibodies which recognize ErbB2 subsets characterized by adjacent tyrosine phosphorylation events (Y1222 and Y1248) in the C-terminal tail of the oncoprotein. Here we use these phosphoantibodies to demonstrate the existence of tyrosine-phosphorylated ErbB2 subsets which differ in their patterns of heterooligomer formation, in vitro autophosphorylation, and recruitment of SH2-containing substrates. Furthermore, Y1222 and/or Y1248 phosphoantibody immunoreactivity is readily detectable in ErbB2-overexpressing human breast tumors, in which context these phosphorylation events exhibit significant discordance. These data confirm the value of site-specific immunodetection as a strategy for characterizing phosphoprotein function in vitro and in vivo and suggest that multisite phosphotyping of human tumors may contribute novel clinicopathologic insights into the significance of the ErbB2 overexpression phenotype.

3T3 Cells↗

Proxy activation of protein ErbB2 by heterologous ligands implies a heterotetrameric mode of receptor tyrosine kinase interaction.

The oncoprotein ErbB2 is frequently overexpressed in human tumours, but no activating ErbB2-specific ligand has yet been identified. Here we analyse the catalytic and oligomeric behaviour of ErbB2 using phosphorylation-state-specific antibodies which distinguish kinase-active and -inactive ErbB2 receptor subsets. Heregulin-alpha (HRG) activates ErbB2 in G8/DHFR 3T3 cells by selectively inducing hetero-oligomerization with kinase-defective ErbB3, indicating that heterologous transphosphorylation is an unlikely prerequisite for ErbB2 activation. HRG also triggers association of epidermal-growth-factor receptors (EGFR) with a kinase-inactive ErbB2 subset while reducing EGFR association with active ErbB2. Similarly, EGF treatment of A431 cells induces concomitant hetero-oligomerization of active ErbB2 with inactive EGFR, of active EGFR with inactive ErbB2, and of inactive ErbB2 with kinase-defective ErbB3. These combinatorial patterns of ligand-dependent oligomerization suggest a multivalent model of receptor tyrosine kinase interaction in which liganded homodimers provide stable oligomerization interfaces for unliganded ErbB2 or other bystander receptors. We submit that ErbB2 may be physiologically activated via a 'proxy' ligand-inducible heterotetrameric mechanism similar to that already established for transforming-growth-factor-beta type I receptors.

3T3 Cells↗

Experimental studies of electroacupuncture on ventricular fibrillation threshold in rats with acute ischemic myocardium.

By ligating the proximate left anterior descend (LAD) of coronary artery and inducing the ventricular fibrillation with electrical stimulation, the preventive effects of electroacupuncture (EA) on ventricular fibrillation were observed. The results showed that the ventricular fibrillation threshold (VFT) of rats with acute ischemic myocardium was raised after acupuncturing some acupoints, which could prevent the occurrence of ventricular fibrillation. Furthermore, the combination of EA and propranolol could enhance the VFT effectively, and they showed a good synergistic effect.

Adrenergic beta-Antagonists↗

Noninvasive assessment of bone density and structure using computed tomography and magnetic resonance.

For several reasons, including low cost and radiation dose, simplicity, and the ability to image several skeletal sites, dual X-ray absoptiometry (DXA) is the most widely employed technique for diagnostic and serial assessment of integral bone mass in osteoporosis and other metabolic bone diseases. However, three-dimensional imaging modalities such as quantitative computed tomography (QCT) and magnetic resonance (MR) imaging offer the ability to separately examine different factors that may play independent and important roles in osteoporosis. These factors include the density of the trabecular and cortical compartments as well as the pattern of trabecular microarchitecture. New developments in QCT include volumetric approaches for precise compartmental assessment of the spine and proximal femur as well as thin-slice tomography of the vertebral body for assessment of trabecular texture. In addition, ultrahigh resolution CT scanners (spatial resolution ë50-150(i)i) have been developed for imaging of trabecular structure in specimens and in some cases for the peripheral skeleton (distal radius and phalanges). High resolution MR measurements may be employed for assessment of the trabecular texture at a range of peripheral sites, including the calcaneus, distal radius, and phalanges.

Adult↗

The duration of phorbol-inducible ErbB2 tyrosine dephosphorylation parallels that of receptor endocytosis rather than threonine-686 phosphorylation: implications for the physiological role of protein kinase C in growth factor receptor signalling.

Tumour cell growth may be accelerated by protein kinase C (PKC) agonists such as phorbol esters and receptor tyrosine kinases, but receptor tyrosine kinases are in turn desensitized to growth factors by PKC agonists. To clarify this apparent PKC bifunctionality, we have used phosphoantibodies to determine the relationship between PKC-dependent phosphorylation events affecting the ErbB2 oncoprotein in G8/DHFR 3T3 cells. Neither the kinetics nor the extent of phorbol-induced juxtamembrane domain (Thr686) phosphorylation vary directly with C-terminal (Tyr1222) dephosphorylation, with Tyr1222 continuing to be dephosphorylated long after Thr686 phosphorylation has also declined. Platelet-derived growth factor (PDGF) mimics the short-term effects of phorbol on Thr686 and Tyr1222 phosphorylation, and confocal microscopy reveals that both of these PKC agonists induce rapid internalization of PKC-modified ErbB2. Phorbol causes sustained cytoplasmic accumulation of PKC-phosphorylated receptors, however, whereas PDGF triggers the appearance of this ErbB2 subset only briefly. Metabolic labelling and co-precipitation studies fail to implicate heterologous molecules in either the tyrosine dephosphorylation or internalization of PKC-modified ErbB2. Taken in the context of earlier juxtamembrane domain mutagenesis studies, these findings indicate that phorbol-activated PKC may desensitize growth factor receptors to extracellular ligands solely by triggering sustained receptor internalization. We submit that PKC-dependent juxtamembrane domain phosphorylation represents a physiological mechanism for shortening the duration and enhancing the specificity of growth factor signalling by promoting internalization of liganded and unliganded receptors, respectively.

3T3 Cells↗

Morphometric texture analysis of spinal trabecular bone structure assessed using orthogonal radiographic projections.

The measurement of bone microstructure as well as bone mineral density may improve the estimation of bone strength. Cubic specimens (N = 26, 12 mm X 12 mm X 12 mm) of human cadaver vertebrae were cut along three orthogonal anatomic orientations, i.e., superior-inferior (SI), medial-lateral (ML), and anterior-posterior (AP). Contact radiographs of the bone cubes along all three orientations were obtained and then digitized by a laser scanner with pixel size of 50 microns x 50 microns. The specimens were tested in compression along the 3 orthogonal orientations and the Young's modulus (YM) was calculated for each direction. Quantitative computed tomography (QCT) was used to obtain a measure of trabecular bone mineral density (BMD). Global gray level thresholding and local thresholding algorithms were used to extract the trabecular bone network. Apparent trabecular bone fraction (ABV/TV), mean intercept length (I.TH), mean intercept separation (I.SP), and number of nodes (N.ND) were measured from the extracted trabecular network. Fractal dimension (Fr.D) of the trabecular bone texture was also measured. Paired t-tests showed that the mean values of each texture parameter (except ABV/TV) and of YM along the SI direction were significantly different (p < 0.05) from those along the ML and AP direction. However, the mean values along the ML and AP directions were not significantly different. Multivariate regression of YM as a function of the texture parameters and BMD showed that without adjusting for the effect of BMD, YM was significantly explained by all the texture parameters (R2 = 0.2-0.6). When BMD was included in the regression, although the variations in YM of ML, AP, and SI orientations could be explained by BMD alone, some of the texture parameters did improve the overall prediction of the biomechanical properties, while, some parameters such as ABV/TV and Fr.D in the ML orientation showed a more significant overall effect in explaining mechanical strength than did BMD. In conclusion, trabecular texture parameters correlated significantly with BMD and YM. Trabecular texture parameters from projectional radiographs reflect the anisotropy of trabecular structure. Quantitative radiographic assessment of trabecular structure using fine-detail radiography can potentially improve the estimation of bone strength.

Adult↗

A comparative study of trabecular bone properties in the spine and femur using high resolution MRI and CT.

The purpose of this study was to use high resolution (HR) magnetic resonance (MR) and computed tomography (CT) images combined with texture analysis to investigate the trabecular structure of human vertebral and femoral specimens and to compare these techniques with bone mineral density (BMD) in the prediction of bone strength. Twenty-nine bone cubes were harvested from 12 proximal femur cadaver specimens and 29 from 8 spines. HR MR and CT images were obtained, and texture analysis techniques were used to assess trabecular structure. Additionally, BMD, elastic modulus (EM), and maximum compressive strength were determined. R2 for EM versus texture measures computed in the MR images was higher (R2 = 0.27-0.64, p < 0.01) in the spine than in the femur specimens (R2 = 0.12-0.22, p < 0.05). R2 values were similar in the CT images. R2 for EM versus BMD was 0.66 (p < 0.01) in the spine and 0.61 (p < 0.01) in the femur specimens. In the MR images, texture measures combined with BMD in a multivariate-regression model significantly increased R2, while improvement was less significant in the CT images. Thus, texture analysis may provide additional information needed to analyze bone strength and quality.

Adult↗

Trabecular bone mineral and calculated structure of human bone specimens scanned by peripheral quantitative computed tomography: relation to biomechanical properties.

The relationship of cortical bone mineral density (BMD), and geometry to bone strength has been well documented. In this study, we used peripheral quantitative computerized tomography (pQCT) to acquire trabecular BMD and high-resolution images of trabeculae from specimens to determine their relationship with biomechanical properties. Fifty-eight human cubic trabecular bone specimens, including 26 from the vertebral bodies, were scanned in water and air. Trabecular structure was quantitated using software developed with Advanced Visual Systems interfaced on a Sun/Sparc Workstation. BMD was also obtained using a whole-body computerized tomography scanner (QCT). Nondestructive testing of the specimens was performed to assess their elastic modulus. QCT and pQCT measurements of BMD of specimens in water were strongly correlated (r2 = 0.95, p < 0.0001), with a slope (0.96) statistically not significantly different from 1. Strong correlations were found between pQCT measurements of specimens in water and in air, for BMD (r2 = 0.96, p < 0.0001), and for apparent trabecular structural parameters (r2 = 0.89-0.93, p < 0.0001). Correlations were moderate between BMD and apparent trabecular structural parameters (r2 = 0.37-0.64, p < 0.0001). Precision as coefficient of variation (CV) and standardized coefficient of variation (SCV) for these measurements was < 5%. For the vertebral specimens, the correlation was higher between elastic modulus and BMD (r2 = 0.76,p < 0.0001) than between elastic modulus and apparent trabecular structural parameters (r2 = 0.58-0.72, p < 0.0001), while the addition of apparent trabecular nodes and branches to BMD in a multivariate regression model significantly increased the correlation with the elastic modulus (r2 = 0.86, p < 0.01). Thus, pQCT can comparably and reproducibly measure trabecular bone mineral in water or air, and trabecular structure can be quantitated from pQCT images. The combination of volumetric BMD with trabecular structural parameters rather than either alone improves the prediction of biomechanical properties. Such a noninvasive approach may be useful for the preclinical study of osteoporosis.

Adult↗

[Relationship between the expression level of alpha-MHC gene and cardiac contractility during heart failure].

To investigate the molecular basis of the decrease in myocardial contractility during heart failure, an animan model of heart failure was set up by means of deoxycorticosterone-acetate impregnated silicone rubber implants in wistar rats. Cardiac contractility in normal and heart failure rats was examined, and gene expression of its myocardial contractile protein, alpha-MHC, was quantitatively analyzed at gene transcription level by using RNA slot blot hybridization. The results showed that the cardiac contractility and the alpha-MHC mRNA levels in heart failure rats were all lower than those in the normal. Statistical analysis showed a positive correlation between the cardiac contractility and the gene expression of alpha-MHC (r = 0.4143, n = 43, P < 0.05). The above results indicate that gene expression level of alpha-MHC is one of the key factors determining cardiac contractility.

Animals↗

[Reversal of cancer multidrug resistance by Chinese medicine Ams-11, Fw-13 and Tul-17].

The aim of this project was to find some kinds of Chinese materia medica as effective agents for in vitro reversal of cancer multidrug resistance. Based on the present authors' previous researches, thirty-two kinds of Chinese medicine as research meterials were selected and examined. Using cell growth inhibit assay, the authors found that three of them--Ams-11, Fw-13 and Tul-17 in the doses free from cytotoxity could enhance the sensitivity of multidrug resistant cells to anti-cancer drugs in a dose-dependent way. It seemed that these three kinds of Chinese medicine might be potential effective reversal agents.

Cell Survival↗

[Qualitative and quantitative analyses of telomerase activity in cultured cells].

In this study, we established a qualitative PCR-TRAP assay by using SYBR Green stain instead of EB stain; this modification based on Kim's method raised the sensitivity of telomerase detection by 25-100 fold. Besides, a quantitative assay was established by us using 32P-ATP labeled TS primer and the internal control TSK1. We detected the telomerase activity of 12 cultured cells by means of these two assays. The results showed that the telomerase activity could be detected in all the cells, but the activity levels of the cells differed prominently (from 23 to 652 TPG units). The establishment of PCR-TRAP assay and especially the establishment of quantitative assay have enabled us to evaluate the telomerase activity of cells more accurately, and they can be used in our further studies of tumor gene therapy by reducing the telomerase activity of tumor cells.

Breast Neoplasms↗

Effects of propranolol on beta-adrenergic receptor of experimental acute myocardial infarction in rats.

By using receptor autoradiography to observe the distribution and density of receptors, the effects of propranolol, a beta-blocker, on beta-adrenergic receptor of experimental acute myocardial infarction (AMI) were studied. One week after ligation of proximate left anterior descend (LAD) coronary artery, [3H] DHA binding sites were markedly decreased in both infarct region and non-infarct region. After treatment of propranolol (100 micrograms/kg), the [3H] DHA binding sites were obviously increased in the infarct region, and they were further decreased in the non-infarct region. The ratio of [3H] DHA binding sites of the infarct region to non-infarct region was from 0.24 at LAD ligation to 0.87 after propranolol treatment, which was close to 0.97 of control group (sham operation). The results indicated that the propranolol acted directly on myocardial beta- adrenergic through the receptor regulation of the balance of beta-receptors between the infarct region and non-infarct region, and improvement of the myocardial consonation and contraction synergism, thereby protecting the heart affected by AMI.

Adrenergic beta-Antagonists↗

High resolution magnetic resonance imaging of the calcaneus: age-related changes in trabecular structure and comparison with dual X-ray absorptiometry measurements.

A high-resolution magnetic resonance imaging (MRI) protocol, together with specialized image processing techniques, was applied to the quantitative measurement of age-related changes in calcaneal trabecular structure. The reproducibility of the technique was assessed and the annual rates of change for several trabecular structure parameters were measured. The MR-derived trabecular parameters were compared with calcaneal bone mineral density (BMD), measured by dual X-ray absorptiometry (DXA) in the same subjects. Sagittal MR images were acquired at 1.5 T in 23 healthy women (mean age: 49.3 +/- 16.6 [SD]), using a three-dimensional gradient echo sequence. Image analysis procedures included internal gray-scale calibration, bone and marrow segmentation, and run-length methods. Three trabecular structure parameters, apparent bone volume (ABV/TV), intercept thickness (I.Th), and intercept separation (I.Sp) were calculated from the MR images. The short- and long-term precision errors (mean %CV) of these measured parameters were in the ranges 1-2% and 3-6%, respectively. Linear regression of the trabecular structure parameters vs. age showed significant correlation: ABV/TV (r2 = 33.7%, P < 0.0037), I.Th (r2 = 26.6%, P < 0.0118), I.Sp (r2 = 28.9%, P < 0.0081). These trends with age were also expressed as annual rates of change: ABV/TV (-0.52%/year), I.Th (-0.33%/year), and I.Sp (0.59%/year). Linear regression analysis also showed significant correlation between the MR-derived trabecular structure parameters and calcaneal BMD values. Although a larger group of subjects is needed to better define the age-related changes in trabecular structure parameters and their relation to BMD, these preliminary results demonstrate that high-resolution MRI may potentially be useful for the quantitative assessment of trabecular structure.

Absorptiometry, Photon↗

Texture analysis of direct magnification radiographs of vertebral specimens: correlation with bone mineral density and biomechanical properties.

RATIONALE AND OBJECTIVES: The authors used direct magnification radiographs, combines with texture analysis, to investigate the trabecular structure of human vertebral specimens and compared these techniques with measurement of bone mineral density (BMD) by using quantitative computed tomography to predict bone strength. METHODS: Direct magnification radiographs and BMD measurements were obtained from 38 motion segments from the thoracolumbar spines of 11 female human cadavers. Maximum compressive strength (MCS) was determined with a materials testing machine. Morphologic parameters, digital skeletons, and fractal dimension were obtained from the radiographs in three different regions of interest. RESULTS: Correlations between BMD and MCS were statistically significant (r = .81, P < .01). With morphologic parameters, correlation coefficients of up to .64 (P < .01) were obtained. Use of multivariate regression analysis with one morphologic parameter (the width of the black pixels, or thicknessB) in addition to BMD improved correlations versus MCS (P < .01). CONCLUSION: In an experimental setting, BMD showed statistically significant correlation with bone strength, whereas the structural parameters demonstrated only modest correlations. BMD together with one of these measures (thicknessB), however, showed the highest correlation.

Adult↗