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Biomedical subjects

B Zhang

Publications and source records attributed to B Zhang.

At least 487 records · Page 27Linked to original sources
PubMed1996

A multiparameter analysis of sickle erythrocytes in patients undergoing hydroxyurea therapy.

During 24 weeks of hydroxyurea treatment, we monitored red blood cell (RBC) parameters in three patients with sickle cell disease, including F-cell and F-reticulocyte profiles, distributions of delay times for intracellular polymerization, sickle erythrocyte adherence to human umbilical vein endothelial cells in a laminar flow chamber, RBC phthalate density profiles, mean corpuscular hemoglobin concentration and cation content, reticulocyte mean corpuscular hemoglobin concentration, 1H-nuclear magnetic resonance transverse relaxation rates of packed RBCs, and plasma membrane lateral and rotational mobilities of band 3 and glycophorins. Hydroxyurea increases the fraction of cells with sufficiently long delay times to escape the microcirculation before polymerization begins. Furthermore, high pretreatment adherence to human umbilical vein endothelial cells of sickle RBCs decreased to normal after only 2 weeks of hydroxyurea treatment, preceding the increase in fetal hemoglobin levels. The lower adhesion of sickle RBCs to endothelium would facilitate escape from the microcirculation before polymerization begins. Hydroxyurea shifted several biochemical and biophysical parameters of sickle erythrocytes toward values observed with hemoglobin SC disease, suggesting that hydroxyurea moderates sickle cell disease toward the milder, but still clinically significant, hemoglobin SC disease. The 50% reduction in sickle crises documented in the Multicenter Study of Hydroxyurea in Sickle Cell Disease is consistent with this degree of erythrocyte improvement.

Adult↗
PubMed1996

Insulin- and mitogen-activated protein kinase-mediated phosphorylation and activation of peroxisome proliferator-activated receptor gamma.

Peroxisome proliferator-activated receptor (PPAR) gamma plays an important role in adipocyte differentiation and the regulation of adipocyte gene expression. Insulin also serves to promote adipogenesis. We report that insulin and a PPARgamma ligand (thiazolidinedione (TZD)) stimulate in a synergistic manner the expression of an adipocyte-specific gene (aP2) in rat adipocytes and 3T3-L1 cells. Potential cross-talk between insulin signaling and PPARgamma was studied in Chinese hamster ovary cells expressing insulin receptors (CHO.T), PPARgamma, and reporter genes. Both TZD and insulin independently stimulated PPARgamma-mediated transactivation of aP2 promoter-luciferase reporter genes; both agents combined resulted in a synergistic effect. Co-transfection of CHO.T cells with dominant-negative mitogen-activated protein (MAP) kinase-kinase (MKK1) abrogated both insulin- and TZD-mediated activation of PPARgamma; transactivation was markedly increased in cells co-transfected with constitutively active MKK1. Both insulin and constitutively active MKK1 also stimulated 32P incorporation into PPARgamma in vivo. The conclusions are: 1) Insulin synergizes with a PPARgamma ligand and can activate the receptor in a ligand-independent fashion. 2) PPARgamma is phosphorylated in vivo by insulin stimulation or activation of the MAP kinase pathway. 3) MAP kinase is an important mediator of cross-talk between insulin signal transduction pathways and PPARgamma function.

3T3 Cells↗
PubMed1996

Defective folding of mutant p16(INK4) proteins encoded by tumor-derived alleles.

p16(INK4) is a specific cyclin D-dependent kinase inhibitor and a multiple tumor suppressor. Inactivation of p16 is frequent in both primary tumors and tumor-derived cell lines. We describe here the conformational properties and oligomerization state of seven mutant p16 proteins; all of them are deficient in function. Four of the seven proteins show significantly disrupted secondary structure and backbone folding. The other three adopt partially folded, molten globule-like conformations. These proteins have near-native levels of secondary structure, but lack the ability to undergo a cooperative thermal transition and are substantially less resistant to proteolysis than is wild type p16. At low concentrations, two of the seven proteins are monomers, three exhibit an apparent molecular weight between the value of a monomer and a dimer, and the other two aggregate significantly. Our results strongly suggest that defective protein folding and/or aggregation is a common mechanism for inactivation of p16.

Alleles↗
PubMed1996

Hypermethylation of the CpG island of p16/CDKN2 correlates with gene inactivation in gliomas.

There is considerable evidence that lack of p16 protein expression is a frequent event in human gliomas. Nevertheless, the molecular mechanisms underlying this absence of p16 protein expression are not completely understood. In some gliomas, homozygous deletions are the main cause of p16/CDKN2 gene inactivation. However, other gliomas lacking p16 expression exhibit intact p16/CDKN2 gene, suggesting that p16/CDKN2 is down-regulated at the transcriptional level. In this study we investigated whether aberrant p16/CDKN2 gene methylation correlated with absence of p16 expression in the latter group of gliomas. In a series of 27 gliomas, 12 malignant tumors exhibited loss of p16/CDKN2 expression but not gene deletion. Methylation analysis of the CpG island in the 5' region of the p16/CDKN2 gene showed that exon 1 was extensively methylated in six and partially methylated in the other six of the 12 malignant gliomas. In contrast, no methylation was observed in four other malignant gliomas and two low-grade gliomas that expressed p16 protein. These results indicate that abnormal hypermethylation of the CpG island encompassing the 5' end of the p16/CDKN2 gene may be a mechanism of transcriptional silencing in gliomas without homozygous deletions.

Brain Neoplasms↗
PubMed1996

Comparison between aortic and sinusoidal liver endothelial cells as targets of hyperacute xenogeneic rejection in the pig to human combination.

Endothelial cells of aortic origin are usually used in vitro as targets of hyperacute xenogeneic rejection, although endothelial cells from organs may have different properties. The sensitivities of aortic and liver endothelial cells to hyperacute xenogeneic rejection were compared in the pig to human combination. Sinusoidal liver endothelial cells were isolated and purified by collagenase perfusion of pig livers, sedimentation on a percoll gradient and selective adherence. Purity and viability of isolated liver endothelial cells after adherence were 85+/-6% and >95%, respectively. Endothelial cells from pig aortae (purity and viability >95%) were isolated by scraping. Immunoblotting analysis of xenoantigens on liver and aortic endothelial cell membranes preparations showed identical patterns. The strongest bands revealed by human IgM were located between 110 and 135 kD, while human IgG detected two major bands at 115 and 75kD. The membrane expression of xenoantigens recognized by human sera, analyzed by flow cytometry, was significantly lower on liver than on aortic endothelial cells (IgM: P=0.0006; IgG: P=0.0009). However, the complement-dependent cytotoxic activity of human sera was the same whether liver (54.5+/-1.4%) or aortic endothelial cells (50.0+/-4.2%) were used as targets. Taken together, those results allow the use of aortic instead of sinusoidal liver endothelial cells in the characterization of pig antigens recognized by human natural antibodies.

Acute Disease↗
PubMed1996

A three-hybrid system to detect RNA-protein interactions in vivo.

RNA-protein interactions are pivotal in fundamental cellular processes such as translation, mRNA processing, early development, and infection by RNA viruses. However, in spite of the central importance of these interactions, few approaches are available to analyze them rapidly in vivo. We describe a yeast genetic method to detect and analyze RNA-protein interactions in which the binding of a bifunctional RNA to each of two hybrid proteins activates transcription of a reporter gene in vivo. We demonstrate that this three-hybrid system enables the rapid, phenotypic detection of specific RNA-protein interactions. As examples, we use the binding of the iron regulatory protein 1 (IRP1) to the iron response element (IRE), and of HIV trans-activator protein (Tat) to the HIV trans-activation response element (TAR) RNA sequence. The three-hybrid assay we describe relies only on the physical properties of the RNA and protein, and not on their natural biological activities; as a result, it may have broad application in the identification of RNA-binding proteins and RNAs, as well as in the detailed analysis of their interactions.

Base Sequence↗
PubMed1996

Down-regulation of the expression of the obese gene by an antidiabetic thiazolidinedione in Zucker diabetic fatty rats and db/db mice.

Obese (ob) is a recently identified gene involved in the regulation of energy balance in the mouse. We report here that AD-5075, a potent thiazolidinedione which lowered plasma glucose and triglyceride in Zucker diabetic fatty (ZDF) rats and db/db mice, decreased the expression of the ob gene in these animal models of obesity and non-insulin-dependent diabetes mellitus. The level of adipose ob mRNA in ZDF rats was 3-fold greater than that detected in the Zucker lean littermates. Chronic treatment with AD-5075 elicited a 67 and 70% reduction of ob mRNA in ZDF and control lean rats, respectively. Furthermore, the amount of adipose ob mRNA in db/db mice was 7 times higher than that detected in lean littermates. Treatment of db/db mice with AD-5075 resulted in a 78% reduction of the level of ob mRNA with parallel changes in circulating level of the ob gene product, leptin. The reduction of the ob mRNA in the Zucker lean rats was accompanied by significantly greater food intake and weight gain. However, in ZDF rats and db/db mice, there was profound increase in body weight without hyperphagia. The results demonstrate that the expression of the ob gene is up-regulated in these two rodent models of diabetes compared to their lean counterparts and that such overexpression is attenuated by treatment with an agent that improves insulin sensitivity and glucose homeostasis in vivo.

Adipose Tissue↗
PubMed1996

Mutations of the p16 gene in gliomas.

In the present study we investigated the frequency of p16 gene exon 2 mutations in 35 malignant gliomas, using either direct sequencing of the PCR products or cloning into the pCRII vector and sequencing of the cloned PCR products. No mutations were detected during direct sequencing of the PCR products. However, after sequencing of individual clones, we found multiple mutations in 5 tumors involving codons 73(GCC to ACC, Ala to Thr), 76 (GCC to GTC, Ala to Val), 85(GCT to ACT, Ala to Thr), 98(CAC to TAC, His to Tyr), 102 (GCG to GTG, Ala to Val), 106 (GTG to ATG, Val to Met), 107 (CGC to TGC, Arg to Cys), 127 (GCA to GTA, Ala to Val), 128 (CGG to TGG, Arg to Trp) and 136 (GGC to GAC, Gly to Asp). Mutations were found only in glioblastomas and were either C to T or G to A transitions. Each mutation was detected in a small percentage of tumor cells (1.3-22%) using individual colony sequencing and southern hybridization with mutant oligonucleotides, consistent with the heterogenous cell population of glioblastomas. The presence of p16 gene mutations only in glioblastomas suggests that they are late events in glioma development.

Adult↗
PubMed1996

Comparison of the antithrombotic effect of the new recombinant hirudin HBW 023 and heparin in small arteries and veins.

The study was composed of two parts, arterial and venous; the 24 rabbits in each arm were divided into three equal groups and treated with either saline (control) or 1 mg/kg body weight (bw) of a new recombinant hirudin HBW 023 given as a single dose or standard heparin 1 mg/kg bw followed by quarter doses of heparin every half hour. Both arms included a control group given equal volumes of saline. The study continued for 2 hours. The following parameters were evaluated: bleeding times from arteriotomy/venotomy, patency rates, and the weights of thrombotic materials. Plasma samples were taken for evaluation of anti-factor lla (anti-Flla), anti-factor Xa (anti-Fxa), and activated partial thromboplastin time (APTT). The bleeding times were significantly prolonged but were still within clinically acceptable levels, following both HBW 023 and heparin treatment. Patency rates were significantly improved in both the arterial and venous arms following HBW 023 and heparin treatment. A corresponding reduction in thrombotic materials was simultaneously registered in the arterial and venous arms following HBW 023 and heparin treatment. Hirudin (HBW 023) significantly improved the reduction compared with the heparin group in the venous study. Heparin treatment caused expected high levels of anti-FXa and prolonged APTT, but hirudin, being at least as effective in antithrombotic potency, changed the pre-treatment levels only slightly. Anti-Flla levels were immediately increased by both heparin and hirudin (the highest levels) but reached low levels after 2 hours of single-dose hirudin treatment, despite a simultaneously excellent antithrombotic effect. We conclude that the new recombinant hirudin HBW 023, like standard heparin, is a highly efficient antithrombotic agent in both small arteries and veins following severe vessel wall trauma. The bleeding times were simultaneously prolonged significantly (still within acceptable limits) following both heparin and HBW 023 treatment in the arterial arm but were only prolonged following heparin treatment in the venous arm. The advantage of r-hirudin HBW 023 was furthermore the single dose administration.

Animals↗
PubMed1996

Influence of early fibrinolysis inhibition on thrombus formation following microvascular trauma.

The effect of the fibrinolysis inhibitor tranexamic acid on early thrombus formation following microvascular trauma was investigated in the central arteries of ears in 86 rabbits (in all 172 vessels), divided into four separate blind randomised studies. In the first part a common end-to-end anastomosis was done and in the last three studies a severe trauma-arteriotomy/intimectomy was performed. Parameters studied were vessel bleeding times, patency rates, weights of intraluminal thrombotic material, haematocrit and plasma fibrinolytic activity. In the first study consisting of 14 control animals and 18 animals treated with 14 mg/kg bw of tranexamic acid, end-to-end anastomosis was performed on the central artery of one ear and on the central vein of the other ear. In the second, third, and fourth studies consisting of 18, 20, and 16 control vessels and the same number of corresponding vessels in treated animals a 7-mm longitudinal arteriotomy followed by a deep 5-mm-long intimectomy was performed. The second and third treated groups were given 14 mg/kg bw of tranexamic acid 5 min and 1 h, respectively, before reflow and the fourth group 28 mg/kg bw 5 min before reflow. The difference between the second and third studies was the addition, to mimic clinical situations, of 8.5 ml saline/kg bw 2 h before reflow in the third study. In conclusion, treatment with a single clinical dose, 14 mg/kg of tranexamic acid, did not influence vessel bleeding times or thrombus formation in the anastomotic or severe trauma models and seems safe to use. Not even a double clinical dose, 28 mg/kg, influenced thrombus formation in a statistically significant way.

Anastomosis, Surgical↗
PubMed1996

Egg production of cockatiels (Nymphicus hollandicus) is influenced by number of eggs in nest after incubation begins.

Cockatiel eggs hatch asynchronously because most eggs are laid after continuous incubation begins. We manipulated numbers of eggs in the nest after cockatiels began incubating to determine whether the number of eggs in the nest influenced clutch size. We also measured plasma prolactin 2, 7, 12, and 17 days after egg laying began to determine whether clutch size covaried with prolactin concentration. We found that total egg production was stimulated in pairs for which the rate of egg accrual was reduced by egg removal, while total egg production was inhibited in pairs for which the egg accrual rate was increased by egg addition. Female, but not male, plasma prolactin concentrations correlated negatively with egg production on day of lay 17 but not before. We conclude that egg production in incubating cockatiels is influenced by feedback from the number of eggs in the nest and that clutch termination may be signaled by rising prolactin levels in females.

Animals↗
PubMed1996

Reaction of metallothionein with ethylenediamine-N,N,N',N'-tetraacetic acid.

The reactions of various rabbit MTs (Cd7Th, Zn7Th, Hg7Th, Bi7Th, Pb7Th) with EDTA were studied at different pH and EDTA concentrations, and were determined to be pseudo-first-order reactions. The reactions were all divided into two steps: one faster and EDTA-dependent, and the other EDTA-independent. The mechanism is also described in detail. Based on obtained results, the varied induction behaviors of MT formation for those metals are discussed.

Animals↗
PubMed1996

Penicillin G-induced microbicidal activity of endothelial cells cultured on gelfoam blocks.

A body of evidence has surfaced documenting the ability of endothelial cells cultured on monolayers to phagocytose but not kill bacteria. Several years ago, a new three-dimensional endothelial cell culturing model was developed, which simulated the morphology of the endothelium in small vessels and capillaries. Given that endothelial cells may be derived from the same pluripotent stem cells as macrophages, the question of whether endothelial cells might phagocytose and kill bacteria was explored. Endothelial cells grown on Gelfoam blocks exhibited bactericidal activity towards Staphylococcus aureus, reaching maximal killing of > 90% after 2 h. Evidence documents the involvement of bacterial adherence to the plasma membrane of the endothelial cell. This is followed by phagocytosis of S. aureus, leading to intracellular killing. Penicillin G, included in the endothelial cell growth medium, was found to be a critical factor in the bactericidal activity demonstrated by Gelfoam blocks laden with endothelial cells.

Animals↗
PubMed1996

Validity of immunological faecal occult blood screening for colorectal cancer: a follow up study.

OBJECTIVES: Immunological faecal occult blood testing has been recognised as a useful method for wide scale screening of colorectal cancer in Japan during the past few years. This study was carried out to evaluate the validity of an immunological faecal occult blood test for colorectal cancer screening. METHODS: A screening programme for colorectal cancer using an immunological faecal occult blood test was conducted on 3365 residents of regions within Nagano prefecture in 1991, and all the subjects were followed up for three years by verification from cancer registration. Based on the incidence of false negative cases among those who had been evaluated as normal by this screening, but were diagnosed as having colorectal cancer, the sensitivity and the specificity of this test were estimated. RESULTS: Four false negative cases were found during the subsequent three year follow up period after screening. Among these four cases, one case was within the first follow up year, one case within the second year, and two cases within the third year. Accordingly, the sensitivity of this test was calculated as 90.9% within one year, 83.3% within two years, and 71.4% within three years, while the specificity was found to be 95.6%, indicating a validity higher than that achieved by a chemical occult blood test. CONCLUSION: These findings suggest that the immunological faecal occult blood test has a high diagnostic accuracy and is a useful strategy for colorectal cancer screening.

Adult↗
PubMed1996

Diagnostic accuracy of immunochemical faecal occult blood test for gastric cancer.

OBJECTIVES: To investigate the accuracy of the immunochemical occult blood test in screening for gastric cancer and to evaluate whether or not the upper digestive tract should be examined when the occult blood test is positive but there is no abnormal sign in the colorectum. METHODS: In a case-control study an occult blood test was carried out on 150 subjects with gastric cancer, 150 subjects with colorectal cancer, and on 300 healthy subjects. Data were analysed from 44,996 persons attending a population screening programme who underwent both radiological gastric cancer screening (barium meal) and colorectal cancer examination by occult blood test. RESULTS: In the case-control study the occult blood test was positive in 27/150 (18%) subjects with gastric cancer, in 112/ 150 (75%) subjects with colorectal cancer, and in 24/300 (8%) healthy controls. In the population screening programme the occult blood test was positive in 4/50 (8%) persons with gastric cancer and 3232/ 44,950 (7%) persons without gastric cancer, indicating no difference between them. CONCLUSIONS: These data show that the immunochemical faecal occult blood test is worthless as a screening test for gastric cancer, and that examination of the upper digestive tract is unnecessary in cases where the faecal occult blood test is positive but there is no sign of colorectal disease.

Case-Control Studies↗
PubMed1996

Characteristics of colorectal cancer with false negative result on immunochemical faecal occult blood test.

OBJECTIVES: A comparative study was carried out to clarify the clinicopathological features of colorectal cancer diagnosed after a false negative result on the immunochemical faecal occult blood test. METHODS: 236 patients with colorectal cancer were studied: 48 patients with negative results and 188 patients with positive results with the faecal occult blood test. The two groups were compared according to their age and sex and by the site, size, macroscopic type, Dukes's classification, and histological type of their cancer lesions. Additionally, the above factors were investigated prospectively and compared in 40 cases of colorectal cancer cases diagnosed as false negative and in matched cases diagnosed as true positive in cancer screening by the immunochemical faecal occult blood test. RESULTS: In both the hospital based case-control study and the screening programme based nested case-control study the prevalence of rectal cancers was higher in the false negative group than in the true positive group (P = 0.02, P = 0.03), but there were no differences between the two groups for any other factors. CONCLUSION: These results suggest that the immunochemical faecal occult blood test is unsuitable for the diagnosis of rectal cancer.

Case-Control Studies↗