Biomedical subjects

J Zhu

Publications and source records attributed to J Zhu.

At least 487 records · Page 27Linked to original sources

PubMed1998

[Comparative molecular field analysis(CoMFA) of allylamine and benzylamine antimycotics].

"Active analog approach" has been employed to search the pharmacophoric conformation of the allylamine and benzylamine antimycotics. A local minimum energy conformation, which is very similar to the crystallographically determined coordinate of naftifine or terbinafine, has been applied to build all compounds. Comparative molecular field analysis (CoMFA) has been used to examine the correlations between the activities against 6 common human pathogenic fungi and the physicochemical properties of 62 allylamine and benzylamine compounds which had been aligned by rms fit rule. The predictive abilities of the resulting 3D-QSAR models have been tested by 8 new synthetic compounds and 5 reported compounds. Finally, we propose an interaction pattern between allylamine, benzylamine antimycotics and the active site of pseudoreceptor based on the results of 2D-QSAR and 3D-QSAR.

Allylamine↗

PubMed1998

[High-level expression of human immunodeficiency virus type 1 gp120 protein in E. coli].

In order to improve protein expression and to produce good and cheap diagnostic antigen, the gene fragment (560bp) in N-terminal of gp120 of HIV-1 LAV strain was amplified by PCR. After digested by EcoR I and Sal I, the fragment was cloned into a high-level expression vector pET28a. The recombinant plasmid pET/120 transfecting BL21 (DE3) produced the protein with high-level expression in the host cell BL21(DE3), which was further proved having good antigenicity and high specificity by indirect ELISA and Western-blot assay. The protein expressed was about 50% of the total bacterial protein by SDS-PAGE electrophoresis test. It was highly expressed in the prokaryotic expression system.

Cloning, Molecular↗

PubMed1998

[Transient response in electric measurement of human auricular point].

This is an experiment on the measuring E approximately t and R approximately t for human auricular point, which shows there is a transient response of electric potential E(t) and voltage drop U(t) when t < 2 tau and there is at intervals change with the time after 2 tau about R(t) and E(t). We have obtained a response function of the E(t) and U(t) by the circuit analysis. The parameter of the transient character is relaxation time tau, tau approximately RC. The mathematical model is in correspondence with the experimental result of the auricular point and TPM simulation. It indicates that the character changing with the time should be extracted after 2 tau. This experiment is of importance to understanding the electric character of the otopoint.

Acupuncture Points↗

PubMed1998

[Transient response in electric measurement of human auricular point].

This is an experiment on the measuring E-t and R-t for human auricular point, which shows there is a transient response of electric potential E(t) and voltage drop U(t) when t < 2 tau and there is at intervals change with the time after 2 tau about R(t) and E(t). We have obtained a response function of the E(t) and U(t) by the circuit analysis. The parameter of the transient character is relaxation time tau,tau approximately RC. The mathematical model is in correspondence with the experimental result of the auricular point and TPM simulation. It indicates that the character changing with the time should be extracted after 2 tau. This experiment is of importance to understanding the electric character of the otopoint.

Acupuncture Points↗

PubMed1998

[Ultrastructures of a mycoplasma-like organism causing mulberry dwarf disease].

The pathogen causing the dwarf disease of mulberry in Tongxiang, Zhejiang Province was investigated based on transmission electronic microscopy of the leaf vein of diseased plants collected in the field. A mycoplasma-like organism was identified in the sieve tube, companion, and parenchyma cells of the phloem tissues infected. The organism was spheric or oval in shape, ranging from 50 nm to 160 nm in diameter, and with bilamina unit membrane of 8-10 nm in thickness, and was filled with nuclear strands or fibrous material. Some nuclei of mesophyll cells in the diseased leaves were found disrupted as their nuclear membrane was broken, followed by of nucleo-cytoplasmic matrix and dispersive disappearance of nucleoli. Furthermore, other ultrastructural aberration including depletion of chloroplast stroma, disruption of chloroplast grana, increase of mitochondria and rough endoplasmic reticulum, and discomposition of some mitochondrial cristae were also observed associated with the pathological changes caused by the mycoplasma-like organism. In contrast, this type of organism was not found in healthy plants examined. Thus, the mycoplasma-like organism is likely to be a pathogen attributable to the mulberry dwarf disease.

Microscopy, Electron↗

PubMed1998

[The comparison on growth rate and photopigments of filaments of Spirulina platensis strain Z with different morphology].

The line-shaped filaments Sp-Z(L) were isolated and cultured from Spirulina platensis Sp-Z. The growth rate of Sp-Z(L) was only 64% as much as that of Sp-Z when the light intensity was 4000 Lux. The contents (x 10(-3) g/g dry weight) of chlorophylls, carotenoids and phycobilins of Sp-Z(L) and Sp-Z were 20.6, 0.343, 5.00 and 24.1 0.297, 4.46, respectively. Moreover, as to the absorption spectra of the three photopigments of Sp-Z(L), red shifts were observed. Therefore, after the spiral Sp-Z breeded or changed into Sp-Z(L), The contents of carotenoids and phycobilins raised 23% and 12%, respectively, but the contents of chlorophylls dropped 14.5%, and the growth rate dropped 36% too. The results probably showed that the decrease of chlorophylls was one of the main reasons that caused the remarkable decrease of the growth rate of Sp-Z(L).

Carotenoids↗

PubMed1998

[The relation of diabetic retinopathy with blood-fat and lipid peroxidation].

PURPOSE: To observe and inquire into the relation of diabetic retinopathy with lipid metabolism, and lipid peroxidation. METHODS: The contents of triglyceride (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C), superoxide dismutase (SOD) and lipid peroxide (LPO) of serum were examined and determined both in thirty patients with diabetes in early stage suffering from retinopathy and twenty four healthy people. RESULTS: TG, TC, LDL-C and LPO in patients group were obviously higher than those in the control group, while HDL-C and SOD were lower than those in healthy control group. CONCLUSION: The disorder of lipid metabolism and the growing intensity of lipid peroxidation caused by diabetes may be one of the dangerous factors to bring about the retinopathy.

Aged↗

PubMed1998

Agonist-induced desensitization and down-regulation of the human kappa opioid receptor expressed in Chinese hamster ovary cells.

In this study, we examined whether the human kappa opioid receptor stably expressed in Chinese hamster ovary cells underwent desensitization and down-regulation after prolonged exposure to the agonist (-)U50,488H. Pretreatment with (-)U50,488H led to a reduction in the magnitude of increase in [35S]GTPgammaS binding by the subsequent application of (-)U50,488H. The extent of desensitization was related to duration of exposure and (-)U50,488H concentration. Pretreatment with (-)U50,488H also reduced the potency of (-)U50,488H in inhibiting forskolin-stimulated adenylate cyclase. In membranes of (-)U50,488H-pretreated cells, the affinity of (-)U50,488H was lower than that in the untreated control, and GTPgammaS had no effect on (-)U50,488H affinity, consistent with the notion of uncoupling of the receptor-G protein complex by (-)U50, 488H treatment. Down-regulation of the kappa opioid receptor also occurred on exposure to (-)U50,488H. Higher (-)U50,488H concentrations and/or longer incubation periods were required for down-regulation than for desensitization. The degree of down-regulation depended on the agonist concentration and incubation time. (-)U50,488H-induced desensitization and down-regulation were blocked by naloxone. (+)U50,488H, an inactive stereoisomer, did not cause desensitization or down-regulation. These results indicate that both processes were receptor-mediated. After incubation with (-)U50,488H and removal of (-)U50,488H, both (-)U50,488H-induced [35S]GTPgammaS binding and receptor number returned to the control level, which indicates that both processes were reversible. Thus, desensitization and down-regulation of the kappa opioid receptor occur after agonist exposure and represent two different adaptation mechanisms.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗

PubMed1997

The relative stabilities of base pair stacking interactions and single mismatches in long RNA measured by temperature gradient gel electrophoresis.

The thermal stability of RNA duplexes differing by a single base pair (bp) substitution or mismatch were investigated by temperature gradient gel electrophoresis (TGGE). All base pair substitutions and mismatches were examined at six sites, and limited changes were investigated at three other sites. DNA templates for in vitro transcription were generated by the polymerase chain reaction (PCR). Transcribed forward and reverse single stranded RNAs were annealed to form 345 bp dupex RNA. Solution melting curves of selected RNAs were in good agreement with the predicted three step transitions. Parallel TGGE was used to determine the relative stabilities of the RNAs, and perpendicular TGGE was employed to obtain mobility transitions and midpoint transition temperatures (Tmu) of the RNAs' first melting domain. The gel solvent included formamide and urea. The Tmu values of the first melting domain were influenced by the identity of the base pair substitution or mismatch as well as by the site's neighboring base pairs. The difference in the transition temperatures (deltaTmu) between pairs of RNA ranged from 0 to 5 degrees C. deltaTmu values were used to determine free energy differences (deltaDeltaG). For RNA pairs distinguished by a base pair substitution, the deltaDeltaG values were closely correlated with free energy differences calculated from stacking free energies determined from melting studies in 1 M Na+ [Serra, M. J., and Turner, D. H. (1995) Methods Enzymol. 259, 242-261.] An algorithm was developed using the free energies of terminal mismatches [Serra, M. J., and Turner, D. H. (1995) Methods Enzymol. 259, 242-261] that provided very good agreement with experimental free energies for the single internal mismatches.

Base Composition↗

PubMed1997

A Study on the Chemical Heterogeneity of Active Surface of Solid Catalysts

A method that can be used to characterize the chemical heterogeneity of active surface of solid catalyst is given. The chemical adsorption behavior of three adsorbates on Ni-W/F-Al2O3-SiO2 catalyst and supported Ni catalyst has been investigated. On the basis of fractal geometry, the roughness fractal dimension (Dr) and the chemisorption fractal dimension (Dc) of two kinds of catalyst are estimated. We use the difference between Dc and Dr to characterize the chemical heterogeneity of active sites on the catalytic surface. As a result, the chemical heterogeneity of the acid sites of Ni-W/F-Al2O3-SiO2 catalyst is very great. As for supported Ni catalyst, the chemical heterogeneity of active sites decreased when impregnating operation times in preparation of the catalyst increased. After impregnated four times, the chemical heterogeneity of its active surface of supported Ni catalyst is eliminated. Copyright 1997 Academic Press. Copyright 1997Academic Press

Journal Article↗

PubMed1997

Analysis of a peptide hormone-receptor interaction in the yeast two-hybrid system.

Interaction between a peptide hormone and extracellular domains of its receptor is a crucial step for initiation of hormone action. We have developed a modification of the yeast two-hybrid system to study this interaction and have used it to characterize the interaction of insulin-like growth factor 1 (IGF-1) with its receptor by using GAL4 transcriptional regulation with a beta-galactosidase assay as readout. In this system, IGF-1 and proIGF-1 bound to the cysteine-rich domain, extracellular domain, or entire IGF-1 proreceptor. This interaction was specific. Thus, proinsulin showed no significant interaction with the IGF-1 receptor, while a chimeric proinsulin containing the C-peptide of IGF-1 had an intermediate interaction, consistent with its affinity for the IGF-1 receptor. Over 2000 IGF-1 mutants were generated by PCR and screened for interaction with the color assay. About 40% showed a strong interaction, 20% showed an intermediate interaction, and 40% give little or no signal. Of 50 mutants that were sequenced, several (Leu-5 --> His, Glu-9 --> Val, Arg-37 --> Gly, and Met-59 --> Leu) appeared to enhance receptor association, others resulted in weaker receptor interaction (Tyr-31 --> Phe and Ile-43 --> Phe), and two gave no detectable signal (Leu-14 --> Arg and Glu-46 --> Ala). Using PCR-based mutagenesis with proinsulin, we also identified a gain of function mutant (proinsulin Leu-17 --> Pro) that allowed for a strong IGF-1-receptor interaction. These data demonstrate that the specificity of the interaction between a hormone and its receptor can be characterized with high efficiency in the two-hybrid system and that novel hormone analogues may be found by this method.

Insulin-Like Growth Factor I↗

PubMed1997

end-1 encodes an apparent GATA factor that specifies the endoderm precursor in Caenorhabditis elegans embryos.

The endoderm in the nematode Caenorhabditis elegans is clonally derived from the E founder cell. We identified a single genomic region (the endoderm-determining region, or EDR) that is required for the production of the entire C. elegans endoderm. In embryos lacking the EDR, the E cell gives rise to ectoderm and mesoderm instead of endoderm and appears to adopt the fate of its cousin, the C founder cell. end-1, a gene from the EDR, restores endoderm production in EDR deficiency homozygotes. end-1 transcripts are first detectable specifically in the E cell, consistent with a direct role for end-1 in endoderm development. The END-1 protein is an apparent zinc finger-containing GATA transcription factor. As GATA factors have been implicated in endoderm development in other animals, our findings suggest that endoderm may be specified by molecularly conserved mechanisms in triploblastic animals. We propose that end-1, the first zygotic gene known to be involved in the specification of germ layer and founder cell identity in C. elegans, may link maternal genes that regulate the establishment of the endoderm to downstream genes responsible for endoderm differentiation.

Amino Acid Sequence↗

PubMed1997

Leukemia-associated retinoic acid receptor alpha fusion partners, PML and PLZF, heterodimerize and colocalize to nuclear bodies.

In acute promyelocytic leukemia (APL), the typical t(15;17) and the rare t(11;17) translocations express, respectively, the PML/RARalpha and PLZF/RARalpha fusion proteins (where RARalpha is retinoic acid receptor alpha). Herein, we demonstrate that the PLZF and PML proteins interact with each other and colocalize onto nuclear bodies (NBs). Furthermore, induction of PML expression by interferons leads to a recruitment of PLZF onto NBs without increase in the levels of the PLZF protein. PML/RARalpha and PLZF/RARalpha localize to the same microspeckled nuclear domains that appear to be common targets for the two fusion proteins in APL. Although PLZF/RARalpha does not affect the localization of PML, PML/RARalpha delocalizes the endogenous PLZF protein in t(15;17)-positive NB4 cells, pointing to a hierarchy in the nuclear targeting of these proteins. Thus, our results unify the molecular pathogenesis of APL with at least two different RARalpha gene translocations and stress the importance of alterations of PLZF and RARalpha nuclear localizations in this disease.

Animals↗

PubMed1997

Tumor treatment by sustained intratumoral release of cisplatin: effects of drug alone and combined with radiation.

PURPOSE: The effect of intratumoral delivery of cisplatin to a mouse tumor model (RIF-1) by means of a biodegradable polymer implant with and without radiation was studied. METHODS AND MATERIALS: The polymer bis (p-carboxyphenoxy) propane-sebacic acid (CPP:SA; 80:20) and its degradation products have been characterized. Polymer rods (8 x 0.5 mm) containing 17% cisplatin by weight were prepared by extrusion, and the in vitro degradation rate measured. The implants were placed into mouse tumors and their effect (with and without radiation) on tumor growth delay studied. The levels of Pt in the mouse kidney, tumor, and blood plasma at selected intervals after implant were also determined. These results were compared with those obtained when cisplatin was delivered systemically. RESULTS: When cisplatin was delivered by the polymer implants, higher levels were present in the tumor for longer time periods (cf. systemic delivery of the drug). For both nonirradiated and irradiated tumors, those treated with the polymer implants had significantly longer tumor growth delays compared to nonimplanted controls and to systematically treated tumors. CONCLUSIONS: The results show that intratumoral delivery of cisplatin is more efficient than systemic delivery. Using the biodegradable polymer implant, higher doses of cisplatin can be tolerated by the animal as the drug is localized within the tumor, and the high levels of the drug in the tumor can be maintained for an extended period of time. When radiation is given in conjunction with cisplatin, the tumor response is supraadditive for all modes of cisplatin administration but is potentiated to a greater extent when cisplatin is delivered through the polymer implant. The greatest effect is seen for treatment with cisplatin delivered by polymer implant combined with fractionated radiation.

Animals↗

PubMed1997

SAGE transcript profiles for p53-dependent growth regulation.

Serial analysis of gene expression (SAGE) allows for a quantitative, representative, and comprehensive profile of gene expression. We have utilized SAGE technology to contrast the differential gene expression profile in rat embryo fibroblast cells producing temperature-sensitive p53 tumor suppressor protein at permissive or non-permissive temperatures. Analysis of approximately 15,000 genes revealed that the expression of 14 genes (P < 0.001, > or = 0.03% abundance) was dependent on functional p53 protein, whereas the expression of three genes was significantly higher in cells producing non-functional p53 protein. Those genes whose expression was increased by functional p53 include RAS, U6 snRNA, cyclin G, EGR-1, and several novel genes. The expression of actin, tubulin, and HSP70 genes was elevated at the non-permissive temperature for p53 function. Interestingly, the expression of several genes was dependent on a non-temperature-sensitive mutant p53 suggesting altered transcription profiles dependent on specific p53 mutant proteins. These results demonstrate the utility of SAGE for rapidly and reproducibly evaluating global transcriptional responses within different cell populations.

Animals↗

PubMed1997

Permanent and specific transplantation tolerance induced by a nonmyeloablative treatment to a wide variety of allogeneic tissues: I. Induction of tolerance by a short course of total lymphoid irradiation and selective elimination of the donor-specific host lymphocytes.

The long-term success of organ transplantation is limited by complications resulting from consistent nonspecific immunosuppression. Induction of stable, donor-specific tolerance remains the main goal of transplantation immunology. In this article, a new, nonmyeloablative method is described for induction of transplantation tolerance to fully mismatched bone marrow cells (BMC), bone marrow stromal precursors, heart muscle, and skin allografts. The method is based on pretransplant conditioning with no postgraft immunosuppression, and consists of a short course (six daily fractions of 200 cGy) of total lymphoid irradiation (sTLI), followed by selective elimination of donor-specific alloreactive cells of the host escaping low-dose sTLI. Donor-specific alloreactive cells were activated by intravenous inoculation with a high dose of donor BMC (3 x 10(7) cells) 1 day after sTLI, and eliminated by a single intraperitoneal dose (200 mg/kg) of cyclophosphamide given 1 day after cell transfer. Infusion of a low number of T cell-depleted BMC (3 x 10(6) cells) after tolerogenic preconditioning converted recipients to stable mixed chimeras free of graft-versus-host disease. The same treatment provided long-lasting acceptance of heterotopically transplanted allografts of the heart muscle and of the stromal precursors to the hematopoietic microenvironment. This treatment also led to acceptance and life-long survival of full-thickness donor skin allografts. However, skin allografts survived only in mice that received donor T cell-depleted BMC after cyclophosphamide and had 20-50% donor cells in the blood. Our results suggest that after sTLI, additional selective clonal deletion of residual host cells induces a state of long-lasting specific tolerance to a wide variety of donor-derived tissues.

Animals↗

PubMed1997

AP-1 cis-response elements are involved in basal expression and Vmw110 transactivation of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10).

The promoter of the large subunit of herpes simplex virus type 2 ribonucleotide reductase (ICP10) has two AP-1 cis-response elements, respectively located at positions -62 and -94 relative to the transcription start site (Wymer et al., 1989. J. Virol. 63, 2773-2784). Chloramphenicol acetyl transferase (CAT) analysis with hybrid constructions of the CAT structural gene and the ICP10 promoter or its mutants and gel retardation studies were used to examine the role of the AP-1 cis-response elements in expression from the ICP10 promoter. Basal expression from the wild-type promoter was significantly (75-90%) reduced by mutation of the upstream or downstream AP-1 element. Mutation in the upstream AP-1 element also caused a 60% reduction in c-Jun-mediated activation. Activation was decreased 40% by mutation in the downstream AP-1 element and it was abrogated by mutation of both elements. Similar results were obtained for ACT-deleted mutants and mutants in which CT was mutated to AG. The trans-activation by Vmw110 was also reduced by mutation of the AP-1 elements (10- and 2-fold for the upstream and downstream element, respectively) and it was abrogated by mutation of both AP-1 elements. Mutation of nucleotides adjacent to the AP-1 cis-response elements had no effect on trans-activation. Gel retardation assays with a DNA probe representing the wild-type ICP10 promoter and nuclear extracts from HSV-1-infected cells identified one complex that was not seen with mock-infected cells or with cells infected with a Vmw110-deleted mutant. The complex was not seen when HSV-1-infected cells were reacted with an AP-1-mutant DNA probe, and its formation was competed by an AP-1 but not a mutant AP-1 oligonucleotide. The migration of this complex was retarded by c-Fos antibody, suggesting that both AP-1 and Vmw110 are involved in its formation. A mutant deleted in all sequences upstream of the TATA box was also activated by Vmw110, but this activation was only 2-fold lower than that seen for the wild type and significantly higher (10-fold) than that seen for the double AP-1 mutants. The data suggest that AP-1 elements play a crucial role in ICP10 gene expression/activation.

Animals↗

PubMed1997

Rad and Rad-related GTPases interact with calmodulin and calmodulin-dependent protein kinase II.

Members of the Rad family of GTPases (including Rad, Gem, and Kir) possess several unique features of unknown function in comparison to other Ras-like proteins, with major N-terminal and C-terminal extensions, a lack of typical prenylation motifs, and several non-conservative changes in the sequence of the GTP binding domain. Here we show that Rad and Gem bind to calmodulin (CaM)-Sepharose in vitro in a calcium-dependent manner and that Rad can be co-immunoprecipitated with CaM in C2C12 cells. The interaction is influenced by the guanine nucleotide binding state of Rad with the GDP-bound form exhibiting 5-fold better binding to CaM than the GTP-bound protein. In addition, the dominant negative mutant of Rad (S105N) which binds GDP, but not GTP, exhibits enhanced binding to CaM in vivo when expressed in C2C12 cells. Peptide competition studies and expression of deletion mutants of Rad localize the binding site for CaM to residues 278-297 at the C terminus of Rad. This domain contains a motif characteristic of a calmodulin-binding region, consisting of numerous basic and hydrophobic residues. In addition, we have identified a second potential regulatory domain in the extended N terminus of Rad which, when removed, decreases Rad protein expression but increases the binding of Rad to CaM. The ability of Rad mutants to bind CaM correlates with their localization in cytoskeletal fractions of C2C12 cells. Immunoprecipitates of calmodulin-dependent protein kinase II, the cellular effector of Ca2+-calmodulin, also contain Rad, and in vitro both Rad and Gem can serve as substrates for this kinase. Thus, the Rad family of GTP-binding proteins possess unique characteristics of binding CaM and calmodulin-dependent protein kinase II, suggesting a role for Rad-like GTPases in calcium activation of serine/threonine kinase cascades.

Amino Acid Sequence↗