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Biomedical subjects

Y Shi

Publications and source records attributed to Y Shi.

At least 613 records · Page 34Linked to original sources
PubMed1993

Yin-yang 1 activates the c-myc promoter.

Previous studies on the murine c-myc promoter demonstrated that a ubiquitously present protein, common factor 1 (CF1), bound at two sites located -260 and -390 bp from the P1 transcription start site. CF1 has been purified to near homogeneity and shown to be identical to the zinc finger protein Yin-yang 1 (YY1) as judged by similarity of molecular weight and other biochemical properties, immunological cross-reactivity, and the ability of recombinant YY1 to bind to CF1 sites. In cotransfection experiments, YY1 is a strong activator of transcription from c-myc promoter-based reporters. Furthermore, in murine erythroleukemia cells, overexpressed YY1 causes increased levels of c-myc mRNA initiated from both major transcription initiation sites of the endogenous c-myc gene.

3T3 Cells↗
PubMed1993

Downregulation of c-myc expression by antisense oligonucleotides inhibits proliferation of human smooth muscle cells.

BACKGROUND: Proliferation of smooth muscle cells (SMCs) plays an important role in vascular pathobiology, being involved in the development of coronary restenosis and atherosclerosis. The activation of nuclear proto-oncogenes appears to be a final common pathway onto which various mitogenic signals coverage. Accordingly, we attempted to determine whether the activation of the c-myc nuclear proto-oncogene is essential for human SMC proliferation and explored the possibility of inhibiting their growth using antisense oligonucleotides directed against c-myc messenger RNA (mRNA). METHODS AND RESULTS: Proliferation of human SMCs was associated with an increase in c-myc mRNA expression after growth stimulation. Using 15-mer phosphorothioate oligonucleotides (oligomers), we tested their growth-inhibitory effect in SMCs in vitro. Antisense oligomers directed against the translation initiation region of the human c-myc gene exhibited a significant antiproliferative effect, whereas sense and mismatched oligomers did not inhibit the growth. The growth-inhibitory effect of c-myc antisense oligomers was dose dependent and preventable by an excess of sense oligomers. Furthermore, growth inhibition of SMCs treated with c-myc antisense oligomers was associated with a marked decrease in the c-myc mRNA level. Phosphorothioate oligomers remained stable in medium containing 20% serum and were detectable in SMCs as early as 1 hour after cell exposure. Intact oligomers rapidly accumulated intracellularly and persisted within human SMCs for at least 16 hours. CONCLUSIONS: c-myc antisense oligomers reduced c-myc expression and produced a significant growth inhibition of human SMCs, indicating an important role of c-myc gene activation in the process of SMC proliferation. Furthermore, extracellular stability and rapid cellular uptake provide the basis for future studies assessing the therapeutic role of the c-myc antisense approach in reducing SMC proliferation in the process of vascular restenosis.

Cell Division↗
PubMed1993

Association between protective efficacy of antibodies to tumor necrosis factor and suppression of nitric oxide production in neonatal rats with fatal infection.

In a rat model of fatal infection caused by Pseudomonas aeruginosa, the circulating level of nitrite/nitrate (NO2-/NO3-), a good indicator for nitric oxide production, was remarkably increased after elevation of circulatory tumor necrosis factor (TNF). Anti-TNF MAb cotreatment was shown to blunt hypoglycemia and hyperlacticemia and was associated with decreased mortality of septic animals. Moreover, anti-TNF MAb significantly reduced not only plasma TNF but also plasma NO2-/NO3- levels. Dexamethasone had a similar effects, and when anti-TNF MAb was used in combination with dexamethasone, the suppression of nitric oxide production and the protective efficacy were more remarkable compared with therapy with either anti-TNF MAb or dexamethasone alone. Our present data suggested that the protective efficacy of anti-TNF MAb may correlate with the suppression of nitric oxide production and also with a modulation in metabolic abnormalities in the septic newborn rats.

Animals↗
PubMed1993

p53 mutations in all stages of thyroid carcinomas.

The p53 gene has been implicated as a tumor suppressor gene whose inactivation by mutations has been noted in a variety of human malignancies. Using single strand conformation polymorphism analysis of cDNA fragments amplified by reverse transcription-polymerase chain reaction, we analyzed 57 thyroid tumor specimens (8 follicular adenomas and 49 carcinomas) for the presence of mutations in exons 5, 6, 7, and 8 of p53 gene. Twelve of 49 (24.5%) of the thyroid carcinomas tested presented a mutated p53 allele, but none of the 8 benign thyroid tumors did. Mutations were found in 1 of 5 anaplastic carcinomas and 11 of 44 differentiated carcinomas. Three of these 11 differentiated tumor specimens showed foci of solid tissue with evidence of dedifferentiation. Two samples (1 with anaplastic carcinoma, the other with papillary carcinoma) had double mutations on the same allele resulting in a frameshift. Most mutations were point mutations, and 50% of those were G:C to A:T transitions. Seventy-five percent of the mutations were in exons 7 and 8. The presence of p53 mutations was not associated with tumor stage or histological type. Our data suggest that p53 mutations are involved in thyroid carcinogenesis and may play an important role in the malignant transformation of thyroid cells as well as thyroid tumor progression.

Adenocarcinoma, Follicular↗
PubMed1993

Differential expression of GAD65 and GAD67 in human, rat, and mouse pancreatic islets.

The smaller form of the autoantigen glutamic acid decarboxylase, GAD65 (formerly the 64,000 M(r) autoantigen), is a major target of humoral autoimmunity in type I diabetes. Human autoantisera have been used extensively to characterize the GAD65 antigen in both rat and human islets, but the protein has escaped detection in mouse islets. We have now analyzed the expression of GAD65 and GAD67, the larger glutamic acid decarboxylase protein, in human, rat, and mouse islets of Langerhans and brain, using human monoclonal islet cell autoantibodies, human autoantisera, and experimentally raised antibodies to glutamic acid decarboxylase. Human monoclonal autoantibodies and experimentally raised antibodies reacted with mouse GAD65 produced in a baculovirus expression system by Western blotting and immunoprecipitation and with GAD65 in mouse brain by immunohistochemistry but failed to detect GAD65 in mouse islets by the latter two methods. However, analysis of mouse islets by Western blotting technique, using the most sensitive experimentally raised antibody, showed that mouse islets express both GAD65 and GAD67 but at levels that are severalfold lower than those in mouse brain or in human and rat islets. Furthermore, both human and rat islets predominantly express GAD65, whereas GAD67 is the major glutamic acid decarboxylase protein in mouse islets. Human islets are significantly distinct from mouse and rat islets and from brain because they only express GAD65, which is consistent with the predominant role of this form as a target of autoantibodies associated with beta-cell destruction in humans. Human as well as rat islet GAD65 are found in both membrane-bound and soluble forms. The low level of glutamic acid decarboxylase expression in mouse islets compared with human and rat islets is likely to have implications for both the development of tolerance to glutamic acid decarboxylase as well as the homing of glutamic acid decarboxylase-specific lymphocytes to the mouse beta-cell. In this context, the results suggest 1) that the mouse is ideal for studies of the consequences of an expression of high levels of glutamic acid decarboxylase in the beta-cell from a transgene and 2) that the rat may be better suited than the mouse for development of nontransgenic animal models of glutamic acid decarboxylase autoimmunity by immunization.

Amino Acid Sequence↗
PubMed1993

[Transformation of recombinant plasmid pRK 41 and amplification of full-length coding region of human brain myelin basic protein cDNA].

Transformation of the recombinant plasmid pRK41 containing 2.15 kd human brain myelin basic protein MBP)-coding sequence and 3' untranslated region (1.2 kb) into the E. coli JM109 was made by using Hanahan's method. Positive colonies were screened with digoxigenin oligo labelled rat brain MBP cDNA fragment (1.2 kb). To remove 3' untranslated region and obtain the full-length coding sequence of MBP cDNA, a pair of specific DNA primers was designed and synthesized. A 600 bp fragment was amplified from the recombinant plasmid, extracted from the positive colony by using polymerase chain reaction (PCR). The PCR fragment was isolated, and then digested with BamH I, Kpn I and BamH I + Kpn I. The results of restriction analysis indicate that the PCR amplified fragment is desirable and can be used directly to constract expression vectors.

Base Sequence↗
PubMed1993

Field evaluation of bednets impregnated with deltamethrin for malaria control.

Trials were undertaken in a hypoendemic area of malaria in an area bordering Vietnam, in Napo County of Guangxi Zhuang Autonomous Region, China. The aim was to compare the relative cost effectiveness of DDT residual spraying and of bednets impregnated with deltamethrin in the malaria control program. The trials were divided into three subgroups: (1) two farming areas and one coal mining area with a total population > 20,000, where the trial consisted of mass bednets impregnated with deltamethrin 15 mg/m2 net surface once a year, (2) one farming area with a population of approximately 3,600 where DDT residual spraying at 2g/m2 was carried out twice a year in May and August; (3) one farming area and one coal mining village with a population of > 4,000 were used as a control. The malaria vector population consisted mainly of Anopheles minimus and An. anthropophagus with a small contribution from An. sinensis. After bednets were impregnated with deltamethrin the mosquitos resting on the surface of the bednets decreased significantly, although there was less effect on the total vector population. The results showed that malaria incidence decreased significantly both in areas where impregnated bednets were used and in areas where residual spraying was undertaken. The positive IFAT rates of residents who slept under impregnated bednets decreased significantly in farming areas, especially in that area where bednet impregnation as a vector control measure had been undertaken for two years, but there was no change in the IFAT rate in DDT sprayed or control areas.(ABSTRACT TRUNCATED AT 250 WORDS)

Bedding and Linens↗
PubMed1993

Structure-function studies of the C-terminal alpha-helix of human interleukin-2 by site-directed mutagenesis.

By using oligonucleotide-mediated site-directed mutagenesis, we obtained three human interleukin-2 analogs with substitution of Pro in the C-terminal amphiphilic alpha-helix, 125Pro-IL-2, 127Pro-IL-2 and 125Pro-127Pro-IL-2. The bioactivities of all these analogs were decreased. The extent of the decrease in bioactivity coincided with the degrees of secondary and tertiary structure destroyed, as evaluated by circular dichroism and intrinsic fluorescence analysis. The results suggested that the integrity of the hydrophobic face in the C terminal alpha-helix is crucial for the structure and function of interleukin-2.

Base Sequence↗
PubMed1993

[Cytogenetic and molecular genetic study on a female with complex translocation of t(Y;15) and t(14;21)].

A 27-year-old female having a baby with Down's syndrome was found to have a translocation t(14;21) and an acrocentric marker chromosome 15. The short arm of the marker was darkly stained in both G- and C-banded preparations, and had the normal Ag-NOR, but showed a homogeneously stained region in the distal part of the marker. By means of QM staining, the variant was found fluorescent, and Y-body like was found in about 9% interphase nuclei, which suggested that there might exist Y chromosome material in genomic DNA of proband. Dot blot hybridization using DNA probe pY3.4 derived from the heterochromatic region on the long arm of Y chromosome demonstrated that the proband carried Y chromosome material in her genome. Further, by in situ hybridization with the same probe, it showed that the variant of the marker had specific autoradiographic silver grains. So, the karyotype of the proband indicated that the combination of conventional banding analysis with chromosome-specific probe was of great significance in studying the micro-aberration of human chromosome.

Adult↗
PubMed1993

[High-resolution cytogenetic study of a patient with Prader-Willi syndrome].

We investigated a patient with Prader-Willi syndrome by using the high-resolution banding technique. His clinical findings were also examined in some detail. The patient had interstitial deletion of 15q, with breakpoints at 15q11.2 and 15q13. However, he had a normal karyotype at about 400 bands per haploid-set. During the course of this study, it was realized that a small deletion in the proximal 15q could be easily overlooked when a mitotic spread of about 400-bands or less per haploid-set was used. It is suggested that the high-resolution banding technique is of great importance in identifying microdeletions and other microaberrations of chromosomes.

Child↗
PubMed1993

[Clinical observation of the effect of pretreatment with nifedipine in preventing oxygen free radicals in the cardiopulmonary release bypass operation].

The effect of pretreatment with nifedipine in preventing oxygen free radicals release was observed in the cardiopulmonary bypass (CPB) operation. Twenty cases were divided into two groups for comparison. Nifedipine (0.6 mg/kg) was given to the test group. MDA, XO, UA and SOD were measured. These examinations were taken at pre-aortic clamping, 15 min after aortic clamping, and 30, 60, 120, 180 min after reperfusion. The results indicated that pretreatment with nifedipine reduced oxygen free radicals release in the CPB operation.

Adolescent↗
PubMed1992

Antisense oligodeoxynucleotides as probes of T-lymphocyte gene function.

Conventional and thiophosphonate-derivatized oligonucleotides were employed to specifically regulate functional gene expression in murine T-cell hybridomas. For example, induction of apoptotic cell death following activation of T-cell hybridomas was examined using antisense oligonucleotides corresponding to several protooncogenes. We found that antisense oligodeoxynucleotides corresponding to c-myc inhibited both the characteristic DNA fragmentation and the loss of cell viability following activation without affecting production of lymphokines. Functional antisense oligonucleotides corresponding to c-fos had no effect in this system. These results demonstrate the use of antisense oligonucleotides to regulate function in T-cell hybridomas and provide valuable insights into the molecular bases of this biological phenomenon. Antisense oligonucleotides were also used to study another problem, the relation of T-cell-derived antigen-specific immunoregulatory factors to the T-cell receptor (TCR). Because the translation start of each TCR gene usually varies from one T cell to another, antisense oligonucleotides corresponding to the TCR V alpha or V beta of different cells were shown to act in a cell-specific manner. Furthermore, this method was used to demonstrate that a soluble antigen-specific regulatory activity produced by one of the T-cell lines depends on expression of the specific TCRa, an observation that has since been confirmed by gene transfer experiments. Expression of the CD3-TCR complex on the cell surface was also blocked by antisense oligonucleotides corresponding to CD3 gamma and CD3 zeta; however, neither these nor TCR V beta antisense oligonucleotides had any effect on production of the soluble regulatory activity.

Animals↗
PubMed1992

Displacement of BrdUrd-induced YY1 by serum response factor activates skeletal alpha-actin transcription in embryonic myoblasts.

Muscle-restricted transcription of the skeletal alpha-actin gene is controlled in part by a positive regulator, serum response factor (SRF), and a negative regulator, F-ACT1, which bind competitively to the most proximal serum response element (SRE1). We show here that F-ACT1 is identical to a transcription factor recently cloned and described as YY1, NF-E1, delta, or UCRBP. We found that although the DNA-binding activity of SRF accumulates during myogenesis, that of YY1 diminishes simultaneously. Myoblasts rendered incapable of differentiation by BrdUrd treatment exhibited the highest level of YY1 and the lowest level of SRF activities. Transfected SRF could directly transactivate the skeletal alpha-actin promoter by overcoming the inhibitory effect of BrdUrd-induced YY1. The transactivation depends on intact SRE DNA elements and requires the DNA-binding/dimerization domain of SRF as well as its C-terminal half rich in serines and threonines. Since the functions of YY1 and SRF appear to be developmentally regulated, the convergence of their binding sites upon the SRE constitutes an integrated mechanism whereby temporal and spatial muscle gene expression may be accomplished.

Actins↗
PubMed1992

Role for c-myc in activation-induced apoptotic cell death in T cell hybridomas.

Immature T cells and some T cell hybridomas undergo apoptotic cell death when activated through the T cell receptor complex, a phenomenon that is probably related to antigen induced negative selection of developing T cells. This activation-induced apoptosis depends on active protein and RNA synthesis in the dying cells, although none of the genes required for this process have previously been identified. Antisense oligonucleotides corresponding to c-myc block the constitutive expression of c-Myc protein in T cell hybridomas and interfere with all aspects of activation-induced apoptosis without affecting lymphokine production in these cells. These data indicate that c-myc expression is a necessary component of activation-induced apoptosis.

Animals↗
PubMed1992

Selective dopamine-1 receptor agonist augments regional myocardial blood flow: comparison of fenoldopam and dopamine.

A new class of vasodilators exhibiting selective dopamine-1 receptor agonist activity is being introduced into clinical practice. Inasmuch as various vasodilators either augment or decrease myocardial blood flow ("coronary steal") depending on their pharmacologic action, the goal of this study was to assess the effects of fenoldopam (selective dopamine-1 receptor agonist) and dopamine (nonselective dopamine-1 receptor agonist) on regional myocardial blood flow in the presence of coronary occlusion. Accordingly, in 16 dogs anesthetized with pentobarbital, the left anterior descending coronary artery was occluded. Cardiovascular and renal hemodynamic effects were measured before and after intravenous infusion of renal equipotent doses of either fenoldopam (n = 9, 0.1 micrograms/kg/min) or dopamine (n = 7, 1 micrograms/kg/min). Both fenoldopam and dopamine caused a significant and comparable increase in renal blood flow. Fenoldopam but not dopamine significantly decreased the calculated peripheral vascular resistance and subsequently increased cardiac output. Dopamine had no effect on regional myocardial blood flow. In contrast, fenoldopam augmented transmural myocardial blood flow in normal (from 114 +/- 10 to 188 +/- 27 ml/100 gm/min, p less than 0.02) and ischemic border myocardium (from 45 +/- 5 to 68 +/- 11 ml/100 gm/min, p less than 0.03 and p less than 0.02 vs dopamine). There was a significant increase in blood flow to both the endocardial and epicardial layers of normal and ischemic border myocardium. These changes were accompanied by a significant reduction in coronary vascular resistance in the normal myocardium.(ABSTRACT TRUNCATED AT 250 WORDS)

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗
PubMed1992

Intracellular distribution of a nuclear localization signal binding protein.

The transport of proteins into the nucleus requires the recognition of a nuclear localization signal sequence. Several proteins that interact with these sequences have been identified, including one of about 66 kDa. We have prepared antibodies that recognize the 66-kDa nuclear localization signal binding protein (NLSBP) and inhibit nuclear localization in vitro. By immunofluorescence, it is seen that the NLSBP is predominantly cytoplasmic and is distributed peripherally around the nucleus and the microtubule organizing center. There is also a weak punctate staining of the surface of the nucleus. Methanol-fixed cells can also be stained directly with fluorescently labeled karyophilic proteins. These stains reveal the same cytoplasmic structures as anti-NLSBP. The expression of the NLSBP is growth dependent. When cells grown to confluence are examined, the cytoplasmic staining is greatly reduced, leaving the punctate nuclear staining as the predominant feature. In serum-starved cells, very little staining of either the cytoplasm or the nucleus can be seen. Upon simulation by the addition of serum, the original cytoplasmic and nuclear envelope staining is restored. Cells grown in the presence of colchicine or taxol have an altered NLSBP distribution but apparently normal cytoplasmic nuclear transport.

Amino Acid Sequence↗