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X Ouyang

Publications and source records attributed to X Ouyang.

50 records · Page 3Linked to original sources

Reduced ability of transforming growth factor-alpha to induce EGF receptor heterodimerization and downregulation suggests a mechanism of oncogenic synergy with ErbB2.

The epidermal growth factor receptor (EGFR) is activated by a variety of ligands including EGF and transforming growth factor-alpha (TGFalpha), whereas no ligand for the homologous ErbB2 oncoprotein has yet been identified. Here we use both an ErbB2 phosphoantibody (aPY1222) and an activation-specific EGFR antibody to show that low concentrations of EGF induce more efficient tyrosine phosphorylation of ErbB2 in A431 cells than does equimolar TGFalpha, while EGFR is more potently activated by TGFalpha. Co-precipitation studies confirm that heterodimerization of activated EGFR and transphosphorylated ErbB2 is readily induced by EGF but not TGFalpha. EGFR downregulation is also more efficiently induced by EGF, suggesting that ligand-dependent modification of ErbB2 may be required to terminate EGFR signalling in cells expressing both receptor types. These findings indicate that EGF and TGFalpha differ in their abilities to induce tyrosine phosphorylation and heterodimerization of ErbB2, and raise the possibility that ErbB2 exerts its oncogenic effect in part by impairing TGFalpha-dependent EGFR downregulation.

Antibodies↗

A new trabecular region of interest for femoral dual X-ray absorptiometry: short-term precision, age-related bone loss, and fracture discrimination compared with current femoral regions of interest.

We defined a new region of interest (ROI) for femoral Ward's triangle, centered on the femoral neck axis including comparatively trabecular-rich bone. Forty-seven premenopausal, 39 healthy postmenopausal, and 35 osteoporotic postmenopausal women with vertebral fractures were evaluated comparing the new with the standard femoral ROIs, using a Hologic QDR-2000. Additionally, spinal dual X-ray absorptiometry (DXA) was performed. The short-term precision error of the new ROI expressed as the root mean square of the coefficient of variations was 1.34% for premenopausal women, 1.69% for healthy postmenopausal women, and 2.46% for osteoporotic postmenopausal women. Bone mineral density (BMD) values of the new ROI correlated highly with those of the standard femoral ROIs (r = 0.91 - 0.96) and the spinal BMD (r = 0.74). Age-related bone loss of the new ROI was 0.75% per year (r = 0.66) in healthy women, which was approximately 1.5 times higher than the bone loss of the standard femoral ROIs, except for Ward's triangle. Regarding the intergroup discrimination, the t-value of the new ROI was similar to the t-value of Ward's triangle, and the intergroup percent decrements in BMD of the new ROI approximated those of Ward's triangle. For discriminating women with vertebral fractures, the new ROI demonstrated odds ratios of 1.6 similar to most ROIs but lower than that of the trochanteric region. The new, substantially trabecular ROI appears to be an alternative to the Ward's ROI traditionally used in femoral DXA having improved short-term precision and comparable sensitivity.

Absorptiometry, Photon↗

[Preparation of chromosomal DNA from whole blood without use of phenol].

We describe in this paper a simple and efficient procedure for preparing peripheral blood DNA without pre-isolation of white blood cells. The method also avoids the hazard of organic solvent extraction. Routinely it yields 20 to 28 micrograms of chromosomal DNA from per ml whole blood. The A260/A280 ratio for DNA sample is 1.80 to 1.85. The undigested DNAs migrate above the 21 kb pair, while the endonucleasedigested samples appear characteristically in the smear form. These data illustrate the consistency and quality of the chromosomal DNA isolated using this procedure. The DNA prepared by this method was used for analysis of beta-thalassemia mutation and prenatal diagnosis in our laboratory.

Blood Chemical Analysis↗

[Serological survey of human T lymphotropic virus type I (HTLV-1) IgG antibody in south region of Xinjiang].

Human T lymphotropic virus type-1 (HTLV-1) is endemic in southwestern Japan, Seychelles Islands, Caribbean basin, Brazil, and Sub Saharan Africa. Recently, the prevalence of HTLV-1 of domestic source has been reported from Beijing, Fujian and so forth. The object of our study is to know whether the prevalence of HTLV-1 is present in Xinjiang. We collected 2642 serm samples of various ages and different nationalities (Uigur, Han and Khalkhas) from south region of Xinjiang and tested for determination of HTLV-1 IgG antibody by IFA. The results showed that the total positive rate of HTLV-1 IgG was 0.34% (9/2642), of which Uigur nationality was 0.74% (8/1082), Khalkhas nationality was 0.21 (1/471), Han nationality was zero (0/1089). This data indicated that there are HTLV-1 infection among the population of Xinjiang and especially in Uigur and Khalkhas. Why did the minority nationalities have relatively high frequency of HTLV-1 infection? This needs to be studied further.

Adolescent↗

Human cancer cells exhibit protein kinase C-dependent c-erbB-2 transmodulation that correlates with phosphatase sensitivity and kinase activity.

The c-erbB-2 receptor tyrosine kinase is often overexpressed in human tumors, but the functional implications of this phenotype remain unclear. We previously used phosphorylation-specific antibodies to define major differences in c-erbB-2 tyrosine kinase activity between overexpressing human tumor cell lines (Epstein, R. J., Druker, B. J., Roberts, T. M., and Stiles, C. D. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 10435-10439). Here we extend this approach to define the relationship between c-erbB-2 tyrosine phosphorylation and protein kinase C (PKC)-dependent transmodulation. Phosphorylation-specific antibodies to the juxtamembrane PKC site Thr686 recognize tyrosine-dephosphorylated wild-type c-erbB-2 following G8/DHFR 3T3 cell treatment with PKC agonists. B104-1-1 cells transformed by activated c-erbB-2 express a subset of tyrosine-phosphorylated receptors that are homologously phosphorylated on Thr686, indicating that Thr686 phosphorylation alone is insufficient to abrogate receptor tyrosine phosphorylation. Similarly, the c-erbB-2-overexpressing human cancer cell lines SK-Ov-3 and BT-474 express constitutively Thr686-phosphorylated receptors. SK-Ov-3 cells express predominantly kinase-inactive c-erbB-2 that is heavily Thr686-phosphorylated, indicating that Thr686 phosphorylation in this line is heterologous in origin. In contrast, BT-474 cells express constitutively autophosphorylated c-erbB-2 despite Thr686 phosphorylation. These results indicate that Thr686 phosphorylation does not directly abolish c-erbB-2 activity and suggest that such phosphorylation reflects constitutive PKC activity induced by either receptor-activating mutations or heterologous growth factors. The latter possibility suggests in turn that c-erbB-2 interacts in an as yet undefined way with heterologous growth factor receptors in human tumor cells.

Animals↗

Synthesis, radiosynthesis, and biological evaluation of fluorinated thienylcyclohexyl piperidine derivatives as potential radiotracers for the NMDA receptor-linked calcium ionophore.

Various thienylcyclohexyl piperidine (TCP) derivatives were synthesized and evaluated as potential candidates for use as radiotracers for the in vivo study of the NMDA receptor ion-channel. Modification of the thienylcyclohexylpiperidine was accomplished by substituting its piperidine ring either with other cycloamine rings or N-alkyl-substituted amines, and these two classes of TCP derivatives were synthesized using two different methods. The compounds exhibited affinities ranging from 65 nM up to micromolar in competition assays for the receptor ion-channel labeled with 3H-(+)-MK-801 in rat brain homogenates. Radiosynthesis of 1-[1-(2-thienyl)-4-([18F]fluoro)-cyclohexyl]-1,2,5, 6-tetrahydropyridine was carried out by nucleophilic substitution reaction of 1-[1-(2-thienyl)-4-tosyloxycyclohexyl]-1,2,5,6-tetrahydropyridine with no carrier added 18F-, and the yield was approximately 5-10% (decay corrected) in specific activities of 500-1000 Ci/mmol after reverse-phase HPLC purification. The tracer showed good uptake in rat brains after i.v. injection (approx. 0.10% injected dose/g at 30 min. p.i.). However, the specific uptake in receptor-rich regions (striata, hippocampus, frontal cortex, and parietal cortex) improved only marginally with time compared to cerebellum. Three hours postinjection, parietal cortex showed a maximum ratio of 1.9. Preliminary PET experiment with this radiotracer in a rhesus monkey showed good uptake in the brain regions. However, little retention of the radiotracer was observed in the receptor-rich regions.

Animals↗

A single amino acid substitution in the capsid protein VP1 of coxsackievirus B3 (CVB3) alters plaque phenotype in Vero cells but not cardiovirulence in a mouse model.

We previously described a large plaque attenuant (p14V-1) derived from a cardiovirulent Coxsackievirus B3 (CVB3) and showed that there were no major determinants of either attenuation or plaque phenotype in the 5' nontranslated region (5'NTR). Part of the region encoding the last 124 amino acids of VP3 and the first 106 amino acids of VP1 of the attenuant was then sequenced and compared to the wild-type. Three nucleotide changes were found in the VP1 coding region: a silent single base change at nucleotide position 2467 (C to U) and a double-base change at position 2690-1 (AA to GT), which leads to a change from lysine to serine at amino acid position 80. This mutation maps to the begining of B-C loop of the three-dimensional structure of VP1 of CVB3, where a distinct surface projection is formed. Two infectious chimeric cDNA clones were constructed, based on a cardiovirulent cDNA construct. In one construct, the 5'NTR and the VP3-VP1 region were from p14V-1 and in the other, only the VP3-VP1 region was from this attenuant. Both chimeric viruses produced large plaques on Vero cell monolayers, similar to p14V-1 but larger than the prototypic cardiovirulent virus. In vivo experiments showed that both chimeric viruses induced myocarditis in a murine model, similar to wild-type virus. We conclude that mutation serine-80 in capsid protein VP1 of p14V-1 is a determinant of the large plaque phenotype but is not responsible for attenuation.

Amino Acid Sequence↗

Detection of Coxsackievirus B3 RNA in mouse myocarditis by nested polymerase chain reaction.

BACKGROUND: A majority of cases of viral myocarditis are associated with group B Coxsackieviruses (CVB) and the persistence of these viruses in the myocardium is associated with the progression of acute myocarditis to chronic dilated cardiomyopathy. A highly sensitive nested polymerase chain reaction (NPCR) is required to study the mechanisms of viral persistence in the myocardium. OBJECTIVES: To develop an enterovirus group-specific NPCR system, to compare it to the reverse-transcription PCR (RT-PCR) plus Southern hybridisation and to investigate the dynamics of viral RNA in a murine model of myocarditis induced by CVB3. STUDY DESIGN: Primers corresponding to the conserved sequences in the 5'-nontranslated region of enteroviruses were designed to ensure a broad specificity. The specificity of PCR products was confirmed by Southern hybridisation. The sensitivity of RT-PCR or NPCR was assessed using reconstructed infected muscle samples. The myocardial samples of the SWR murine model of CVB3-myocarditis were collected from day 1 to 30 after infection. The presence of viral RNA was detected by the RT-PCR or NPCR and infectious virus was isolated by cell culture. RESULTS: Both RT-PCR and NPCR could detect all 11 representative enteroviruses. The NPCR could detect as few as 0.01 plaque forming unit of virus, 100 times more sensitive than the RT-PCR. Virus was isolated from the myocardium in acute phase, but was no longer recoverable after 9 days. Viral RNA was detected by the NPCR technique throughout the studied period. CONCLUSIONS: An enterovirus group-specific NPCR system was developed and was much more sensitive than the RT-PCR technique. It can replace the Southern hybridisation of RT-PCR products. The presence of viral RNA in the myocardium after acute phase indicates a possibility of CVB3 shifting to persistent infection in the SWR mice.

Journal Article↗

[Influences of capsaicin and electroacupuncture at "point" on acetylcholine (ACh) release in the spinal dorsal horn of rats].

The dynamic changes of ACh activity in the spinal dorsal horn of rats were recorded by ACh ion selective microelectrodes. The influence of applying capsaicin to the spinal surface, administering anti-SP surum to the interior of dorsal horn, cutting off dorsal roots and electroacupuncturing at "points" on the ACh activity were studied. The results showed that electroacupuncture at "points" and capsaicin could facilitate ACh release of the spinal dorsal horn, it was fully inhibited by cutting off dorsal roots or obviously inhibited by administering anti-SP surum. The above results indicated that ACh might participate in the primary afferent of painsensation and acupuncture sensation as a neurotransmitter and SP might adjust the release of ACh in spinal dorsal horn.

Acetylcholine↗

[Analysis of beta-thalassemia mutations and prenated diagnosis in Chengdu population].

Ninety-five of non-differential diagnostic patients were detected by dot-blot analysis on enzymatically amplified DNA with a number of allele specific oligonucleotide probes complementary to the most common mutations in Chengdu population. Prenatal diagnosis was accomplished by the same procedure on enzymatically amplified amniocyte DNA. The result revealed fifty-eight cases of beta-thalassemia. Of the 73 chromosomes tested, twenty-eight (38.4%) had the codon 17(A-->T) mutation, twenty-one (28.8%) had the codon 41-42(-TTCT) mutation, fourteen (19.0%) had the IVS-I-654(C-->T) mutation; nt--28(A-->G) and nt--29(A-->G) mutations were six (8.2%) and four (5.5%) respectively.

Base Sequence↗

Characterization of a murine model of myocarditis induced by a reactivated coxsackievirus B3.

A transfection-reactivated Coxsackievirus B3 (rCVB3), from a full-length cDNA clone of Nancy strain, has previously been shown to be as cardiovirulent as the wild-type virus. Myocarditis induced by this genetically defined virus was compared in SWR mice with the traditional Balb/c model. SWR mice inoculated with rCVB3 developed more severe myocarditis but less severe pancreatitis than Balb/c mice. In contrast to the poor general health and frequent mortality of Balb/c mice following CVB3 infection, the body weight of SWR mice was not affected by CVB3 inoculation and no mortality occurred at titres of 10(2)-10(7) plaque forming units (PFU). Typical myocarditis developed in SWR mice 7 days post infection. Myocarditic foci consisting of necrotic myocardial fibres and mononuclear cell infiltrates resolved by day 30, similar to that observed in Balb/c. However, SWR mice were more sensitive to rCVB3-induced myocarditis than were Balb/c mice: mild myocarditis was induced (4/4) by as low as 10(2) PFU of the virus (ID50 < 10(1.5) PFU), and more severe myocarditis was seen at higher PFU of virus in a dose-dependent manner. The SWR model was tested with attenuated variants derived from cardiovirulent rCVB3. The ID50 for myocarditis was 10(7) PFU for a large plaque-size attenuant and 10(6) PFU for a minute plaque-size attenuant, indicating loss of cardiovirulence by a factor of more than 10(4)-10(5). rCVB3-induced SWR mouse is a sensitive and reliable model for myocarditis. It is useful in assessing the cardiovirulence of different CVB3 variants and evaluating the efficacies of anti-viral therapies. It will allow follow-up study after high dose infection with cardiovirulent rCVB3.

Animals↗

Assessment of trabecular structure using high resolution CT images and texture analysis.

PURPOSE: Our goal was to use high resolution (HR) CT images combined with texture analysis to investigate the trabecular structure of human vertebral specimens and to compare these techniques with bone mineral density (BMD) in the prediction of bone strength. METHOD: HR CT images with a slice thickness of 1 mm were obtained of 28 bone cubes. Four different groups of texture analysis techniques were used to assess these images. In addition, quantitative CT (QCT) was performed and elastic modulus (EM) was determined biomechanically. RESULTS: R2 between EM and BMD was 0.78 (p < 0.01). R2 values for EM versus most of the texture measures were also significant. Texture measures in addition to measures of BMD in a multivariate regression model significantly increased R2 up to 0.87. CONCLUSION: In an experimental setting, texture parameters calculated using HR CT images correlated significantly with EM. Combining texture measures with BMD improved the prediction of EM significantly.

Adult↗