Biomedical subjects

Y Ding

Publications and source records attributed to Y Ding.

At least 235 records · Page 13Linked to original sources

PubMed1997

Release and activation of human neutrophil matrix metallo- and serine proteinases during phagocytosis of Fusobacterium nucleatum, Porphyromonas gingivalis and Treponema denticola.

The phagocytic ingestion of reference strains and clinical isolates of Fusobacterium nucleatum, Porphyromonas gingivalis, and Treponema denticola by polymorphonuclear leukocytes (PMNs) and the concomitant release of PMN granule proteinases were studied by specific functional and immunological assays. PMNs were incubated with the microorganisms anaerobically at 37 degrees C for indicated time periods. The suspensions and pellets were used for phagocytic ingestion assay and electron microscopic study, respectively. The supernatants were used for the measurements of the amounts and activities of the released PMN enzymes including PMN gelatinase (MMP-9), collagenase (MMP-8), serine proteases (elastase and cathepsin G), and lactate dehydrogenase (LDH). Both fluorescence microscopy and transmission electron microscopy showed that F. nucleatum, P. gingivalis and T. denticola were ingested by the PMNs in comparable numbers. However, measurements of the enzymes released from the triggered PMNs revealed major differences among the three species. High amount of elastase was released from the PMNs triggered by F. nucleatum, but not by P. gingivalis or T. denticola. The treatment of PMNs with P. gingivalis whole cells resulted in the release of gelatinase partly in the 82 kD active form, suggesting proteolytic activation of the degranulated 92 kD proMMP-9. The 82 kD active form of gelatinase was not detected upon triggering the PMNs with F. nucleatum and T. denticola. The PMN-bacteria interaction did not result in release of LDH from triggered PMNs indicating the proteinase release was not due to the PMN cell death. The results show that the susceptibilities of the 3 potentially periodontopathogenic microorganisms, F. nucleatum, P. gingivalis and T. denticola to phagocytic ingestion are not directly related to the amounts and activities of PMN enzymes released during the bacteria-PMN interactions. As PMN degranulation is considered as one of the major pathogenic mechanisms in periodontitis, the observed differences among the microorganisms may be important virulence characteristics of these species.

Bacteriological Techniques↗

PubMed1997

The physical location of genes cdc2 and prh1 in maize (Zea mays L.).

A biotin-labelling in situ hybridization technique was first used to physically map two single copy genes, cdc2 and prh1, in maize. These two genes are metabolically interrelated genes. The full-length cDNA clones cdc2ZmA and ZmPPI of genes cdc2 and prh1 were adopted as the probes. They are 1.3 and 1.6 kb in size, respectively. Clone cdc2ZmA was physically mapped on the long arm of chromosomes 4, 8, and 9. The percent distances from centromere to detection site were 57.9 +/- 2.7, 28.4 +/- 1.5, and 88.2 +/- 3.3. The detection rate was 19.2%. Clone ZmPPI was physically mapped on the long arm of chromosomes 4, 6, and 8. The percent distances were 53.6 +/- 1.2, 60.8 +/- 2.9 and 17.1 +/- 1.6. The detection ratio was 18.5%. The technique of chromosome ISH and the relationship between the location and function of these two genes have been discussed.

CDC2 Protein Kinase↗

PubMed1997

nfi, the gene for endonuclease V in Escherichia coli K-12.

Endonuclease V is specific for single-stranded DNA or for duplex DNA that contains uracil or that is damaged by a variety of agents (B. Demple and S. Linn, J. Biol. Chem. 257:2848-2855, 1982). Thus, it may be a versatile DNA repair enzyme. The protein was purified to apparent homogeneity, and from its N-terminal sequence, its gene, nfi, was identified. nfi is immediately downstream of hemE, at kb 4208 (90.4 min) on the current chromosomal map of Escherichia coli K-12. This region was cloned, and plasmid insertion and deletion mutants were used to study its molecular organization. Although nfi is the third of four closely spaced, codirectional genes, it is expressed independently.

Base Sequence↗

PubMed1997

Lead-induced hypertension: interplay of nitric oxide and reactive oxygen species.

An elevation of mean blood pressure was found in rats treated with low lead (0.01% lead acetate) for 3 months, as contrasted to paired Sprague-Dawley control rats. In these rats, measurement of plasma and urine endothelins-1 and -3 revealed that plasma concentration and urinary excretion of endothelin-3 increased significantly after 3 months (plasma: lead group, 31.8+/-2.2, versus controls, 23.0+1.7 pg/mL, P<.001; urinary excretion: lead group, 46.6+11.7, versus controls, 35.6+6.7 pg/24 h, P<.05), whereas endothelin-1 was unaffected. Plasma and urinary nitric oxide (NO) and cyclic GMP concentrations were not significantly changed. However, assay of plasma and kidney cortex malondialdehyde by high-pressure liquid chromatography, as a measure of reactive oxygen species, was elevated in lead-treated rats compared with that in control rats (plasma: lead group, 4.74+1.27, versus controls, 2.14+.49 micromol/L, P<.001; kidney cortex: lead group, 28.75+3.46, versus controls, 16.38+2.37 nmol/g wet weight, P<.001). There was increased NO synthase activity in lead-treated rat brain cortex and cerebellum. In lead-treated rat kidney cortex, the endothelial constitutive NO synthase protein mass was unaffected, whereas the inducible NO synthase protein mass was increased. These data suggest a balance between increased NO synthesis and degradation (by reactive oxygen species) in lead-treated rats, which results in normal levels of NO. Thus, the hypertension may be related to an increase in the pressure substances, endothelin-3 and reactive oxygen species, rather than to an absolute decrease in nitric NO.

Animals↗

PubMed1997

Human neutrophil collagenase MMP-8 in peri-implant sulcus fluid and its inhibition by clodronate.

The exact molecular mechanisms of the loosening of a dental implant are not well-known. The characteristics of implant sulci are similar to those of periodontal sulci regarding gingival crevicular fluid (GCF) and peri-implant sulcular fluid (PISF). Proteolytic enzymes, matrix metalloproteinases (MMPs), participate in peri-implant tissue remodeling. Clodronate is a well-tolerated bisphosphonate-group drug currently used in bone-resorption-related diseases in humans. The mechanisms of bisphosphonate action are not clarified. Collagenase activity in diseased PISF was significantly higher than in the clinically healthy group. Immunoblotting disclosed that diseased PISF contained increased immunoreactives MMP-8 compared with the healthy PISF. The residual latent collagenase activity in the diseased PISF was activated by gold thioglucose and inhibited completely by 100 microM of doxycycline closely resembling pure neutrophil collagenase (MMP-8). The presence of MMP-8 in diseased but not in clinically healthy PISF may prove to be a useful biochemical indicator to monitor peri-implant health and disease. Pure human neutrophil collagenase (MMP-8) and the MMP-8 present in PISF and in the GCF of both loosening implants and periodontitis-affected teeth were efficiently inhibited in vitro by clodronate (50% inhibition [IC50] was achieved by 150 microM of clodronate), an osteoactive, antiresorptive bisphosphonate. Furthermore, the new finding suggests an extended and hitherto-undescribed potential for clodronate in preventing the loosening of both implants and teeth, based on a dual beneficial effect: prevention of both bone resorption/osteolysis and of soft tissue/dental ligament destruction. Potential new therapeutic indications based on the collagenase-inhibiting effect of clodronate provide potential new therapeutic indications for a variety of diseased involving connective tissue breakdown, such as periodontal disease, arthritides, and tumor invasion.

Adult↗

PubMed1997

Pharmacokinetic-pharmacodynamic modeling of metoprolol stereoisomers in spontaneously hypertensive rat.

AIM: To study the combined pharmacokinetic-pharmacodynamic (PK-PD) model of metoprolol stereoisomers, and compare their inhibitory effects on cardiovascular system in the spontaneously hypertensive rats (SHR). METHODS: The drug concentration in plasma was measured by the reversed phase HPLC and the drug effects were recorded by polygraph. The pharmacokinetic parameters and the PK-PD model parameters were calculated. RESULTS: The plasma concentration-time profiles were adequately described by two-compartment model. Differences of Vd between (+)-Met and (-)-Met were found. The relationships between effects and concentration of effect compartment were represented by the sigmoid-Emax model. The Css50 of Vmax, dp/dtmax, and HR inhibitory effects of (+)-Met were larger than those of (-)-Met. CONCLUSION: Stereo-selective drug distribution and different potencies of the inhibitory effects of (+)-Met and (-)-Met existed in SHR.

Adrenergic beta-Antagonists↗

PubMed1997

[Clinical obeservation of improved passing peritoneum extraperitoneal cesarean sectron].

One hundred and fifty puerperas who had indications for cesarean section and factors associated with intrauterine infection were random divided into three groups and adopted three different operative proccdures which were improved passing peritoneum extraperitoneal, extraperitoneal and low segement cesarean section respectively. Three were 50 puerperas in each group, seven paremeters were observed during and after operation. The results showed that the mean times of three groups from cutting skin to fetal head delivered were 19.5, 22.6 and 19.9 minutes respectively (P < 0.05), the mean operative times were 58.04, 68.2 and 72.0 minutes in proper urder (P < 0.01). Gastrointestinal function recovery mean times were 26.6, 26.0 and 47.9 hours (P < 0.01) in sequence. Postoperative morhidity rates were 10%, 12% and 24% (P < 0.01) in order. Postoperative comlicatins were 0%, 26% and 6% (P < 0.01). There were no significant difference both in newborn one minute Apgar grading and blood loss volume during the operation in three groups. Improved passing peritoneun extraperitoneal cesarean section is a simple, convenient, safe and practical oprative method. It is especially applied to those cases who have a bad form of uterine hypomere and factors of intrauterine infection.

Adult↗

PubMed1997

[Efficacy of dihydroartemisinin in treatment of 37 malaria cases].

Dihydroartemisinin is an efficacious derivative of artemisinin which can be taken orally in treatment of malaria with less side reactions. Thirty-seven cases with malaria were treated with this drug in Beijing and in Ruili City, Yunnan Province. Out of them, 25 cases suffered from falciparum malaria with an average parasitemia of 73,218/microliter and 12 from vivax malaria with an average parasitemia of 4,950/microliter. The dosage was 60 mg daily or 2 mg/kg for paediatric patients for 7 successive days with the first dose doubled. All the patients were clinically cured after treatment. Fever subsided 22-72 hours after the beginning of treatment in falciparum malaria patients with an average fever clearance time of 36.24 +/- 15.30 hours and their parasite clearance time was 24-72 hours with an average parasite clearance time of 44.80 +/- 19.09 hours. One of them had recrudescence 19 days after the disappearance of parasites, hence the recrudescence rate of falciparum malaria was 4.8%. Five early cerebral cases in this series were completely cured after the treatment. All the 12 cases with vivax malaria were also clinically cured after treatment, but 1 case had relapse 35 days after treatment. This patient was completely cured with another course of dihydroartemisinin combined with primaquine. All patients tolerated well the oral administration of dihydroartemisinin and no significant side reactions were seen.

Adolescent↗

PubMed1997

[Drug resistance and its mechanism of intrinsic drug-resistant cell line GRC-1].

In order to probe the characteristics of drug resistance and its mechanisms of renal cell carcinoma, drug-resistant spectrum of renal cell carcinoma cell line GRC-1 was detected by in vitro MTT colorimetric assay, the mechanism of drug resistance in GRC-1 was also studied by the methods of both immunocytochemistry assay and flow fluorescence cytometry. The results demonstrated that GRC-1 was cross-resistant to adriamycin, vincrinstine, etoposide and carboplatinium, both mdr1 gene product P-glycoprotein and GST-pi which was an isozyme of glutathione S-transferases were expressed in GRC-1. The accumulation of net intracellular drugs of GRC-1 was less than that of drug sensitive breast cancer cell line MCF7, and the ability of pumping drugs out of cells was higher than that of MCF7. The results suggested that there is an intrinsic multidrug resistance in GRC-1 cell line, and both P-glycoprotein and glutathione systems play a role in the development of drug resistance for GRC-1. GRC-1 is an ideal target cell line for the study of drug resistance.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1997

[Clinico-transcranial Doppler sonography monitoring on vasospasm and delayed cerebral ischemia after resection of intracranial tumors].

The occurrence of vasospasm and delayed cerebral ischemia after resection of intracranial tumor has not received extensive attention clinically, and is often misdiagnosed and improperly treated as surgical brain damage or brain swelling. Seventy-two patients with intracranial tumor were continuously monitored pre- and postoperatively by means of neurological assessment and transcranial Doppler sonography. Vasospasm was found in 35 (48.6%) patients (18 mild, 13 moderate and 4 severe vasospasm). No significant difference among age, sex, surgical approaches, pathological diagnosis, duration of surgery, amount of blood loss and transfusion during surgery were found, but significant difference was seen in cisternal hemorrhage on CT scan and the amount of blood in cerebrospinal fluid. The cause and features of postoperative vasospasm were discussed, transcranial Doppler sonography played an important role in the diagnosis of vasospasm. To decrease the amount of blood in basal cistern by microsurgery in preventing vasospasm and to differentiate vasospasm from brain swelling are helpful to confirm the coexistent or causal relation based on neurological assessment, CT imagine, transcranial Doppler sonography and ICP monitoring both in deciding therapeutic strategy and successfully controlling vasospasm. Nimotop played a key role in preventing brain damage from vasospasm and cerebral swelling.

Adenoma↗

PubMed1997

An experimental study on application of PCR in detection of kala-azar.

A polymerase chain reaction (PCR) technique was developed to detect Kala-azar. Out of 25 patients, 24 were detected, a sensitivity of 96%. The specificity rate, false positive rate, false negative rate, coincidence rate and Youdent's index were 100% (60/60), 0, 4%, 99% and 0.96 respectively. It was shown that the PCR technique can directly detect Kala-azar in samples (1 microliter) from peripheral blood.

Animals↗

PubMed1996

Characterization of single-stranded cAMP response element binding protein (ssCRE-BP) from mouse cerebellum.

It has been speculated that opiate tolerance and dependence may occur at the level of gene expression. Our previous studies have shown that the binding activity of a nuclear factor (ssCRE-BP) to single-stranded CRE of somatostatin gene is altered by long-term treatment with morphine in the mouse cerebellum. ssCRE-BP was purified from the mouse cerebellum by a combination of chromatography on DNA affinity agarose and Mono Q HR. The native protein exhibited a molecular size of 110-150 kDa by gel filtration, and two polypeptides of about 35-40 kDa were observed on SDS-PAGE. The cloning and sequencing of a cDNA encoding ssCRE-BP showed that the protein possesses a glycine-rich domain and a glutamine-rich domain in the amino terminus and the carboxyl terminus, respectively. To investigate the function of ssCRE-BP in the brain, recombinant glutathione-S-transferase (GST) fusion proteins containing ssCRE-BP were expressed in bacterial systems. Rabbit anti-ssCRE-BP antibodies were raised against a GST-ssCRE-BP fusion protein. Using the antibodies in western blot analysis, a polypeptide of approximately 66 kDa was detected in the brain. These findings indicate that ssCRE-BP is involved in opiate tolerance and dependence.

Animals↗

PubMed1996

Gene transfer of transforming growth factor-beta 1 prolongs murine cardiac allograft survival by inhibiting cell-mediated immunity.

Delivery of immunosuppressants directly to allografts using gene transfer and gene therapy approaches may inhibit immune activation while avoiding the systemic toxicity of conventional immunosuppression. Cardiac grafts from allogeneic (C57BL/6, H-2b) donors were transplanted into CBA/J (H-2k) recipients in a heterotopic, non-vascularized model pSVTGF-beta 1, a plasmid encoding murine transforming growth factor-beta 1 (TGF-beta 1) under the control of an SV40 promoter, was directly injected into grafts at surgery and prolonged survival from 12.0 +/- 0.7 to 25.1 +/- 2.1 days (p < 0.001) in a dose-dependent manner. Plasmid gene transfer-induced immunosuppression was localized to the area of the graft because plasmid injected remote from the graft did not prolong allograft survival and systemic immunity was not influenced by local gene transfer. Limiting dilution analysis of graft-infiltrating cells demonstrated that gene transfer reduced the precursor frequency of donor-specific cytotoxic T lymphocytes (CTL) and activated and total interleukin-2 (IL-2) producing helper T lymphocytes (HTL) in graft-infiltrating cells, whereas CTL generation and HTL precursor frequency in splenic lymphocytes were not altered. Additional data revealed that gene transfer inhibited the priming of TH0 cells and the conversion of primed TH1 cells to activated cells without the participation of TH2 suppressors. These data demonstrate that gene transfer of plasmid DNA encoding TGF-beta 1 in vivo suppresses local T cell immunity, which prolongs allograft survival.

Animals↗

PubMed1996

Blockade of multiple costimulatory receptors induces hyporesponsiveness: inhibition of CD2 plus CD28 pathways.

T-lymphocyte activation requires engagement of the T cell receptor with antigen-major histocompatibility complex, and simultaneous ligation of costimulatory pathways via the lymphocyte receptors CD2 and CD28/ CTLA4. Anti-CD2 monoclonal antibody (mAb) blocks the interaction of the antigen-presenting cell receptor CD48 with its ligand CD2, whereas CTLA4Ig binds with high affinity to the antigen-presenting cell ligands B7-1 and B7-2, blocking their interaction with CD28/CTLA4. We tested the immunosuppressive effects of simultaneously blocking both costimulatory pathways. Using donor C57BL/6J (H2b) hearts transplanted to CBA/J (H2k) recipients, anti-CD2 mAb plus CTLA4Ig administered at the time of transplantation prolonged cardiac allograft mean survival time to >120 days compared with untreated controls (12.2+/-0.5 days, P<0.01), anti-CD2 mAb alone (24.8+/-1.0 days, P<0.01), or CTLA4Ig alone (55.0+/-2.0 days, P<0.01). Retransplantation of these recipients with donor-specific and third-party grafts demonstrated that hyporesponsiveness and tolerance were achieved. In vitro stimulation of lymphocytes from tolerant recipients with donor-specific alloantigen resulted in normal cytotoxic T lymphocyte and mixed lymphocyte reaction responses, showing that clonal deletion or anergy did not occur, but that graft adaptation or suppression likely helped to maintain long-term graft survival. In vitro combinations of anti-CD2 mAb and CTLA4Ig suppressed the generation of allogeneic cytotoxic T lymphocytes (58%) and the mixed lymphocyte reaction (36%); CTLA4Ig was more effective in this regard and the two agents were not synergistic. Anti-CD2 mAb and CTLA4Ig suppressed mitogen-driven proliferation in differential fashions, suggesting that they affected independent signaling pathways. Anti-CD2 mAb and CTLA4Ig also inhibited interleukin (IL)-2, IL-4, and IL-2 receptor (CD25). These data indicate that anti-CD2 mAb plus CTLA4Ig induces hyporesponsiveness and tolerance. The mechanism is likely related to the initial disruption of independent pathways of T-lymphocyte activation leading to antigen-specific long-term graft survival.

Abatacept↗

PubMed1996

Mapping of ankyrin binding determinants on the erythroid anion exchanger, AE1.

The association of ankyrin with the AE1 anion exchanger contributes an essential function to the mechanical and viscoelastic properties of the erythrocyte and constitutes the best understood link between the plasma membrane and the underlying membrane skeleton. The AE1 binding domain of ankyrin consists of 24 tandem repeats of a 33-amino acid motif that is present on a wide variety of otherwise unrelated proteins. The experiments described in this paper are aimed at identifying the specific amino acid sequences in AE1 that comprise the ankyrin binding site. We have exploited a cell-free binding assay to quantify the binding affinity of anion exchangers and a recombinant fragment of ANK1, R13-H. Our previous study (Ding, Y., Casey, J. R. and Kopito, R. R. (1995) J. Biol. Chem. 269, 32201-32208) identified an essential role of the amino-terminal 79 AE1 residues in ankyrin binding. The present study extends these findings to show that these 79 amino acids, although necessary, are not sufficient for ankyrin binding. Using chimeras between AE1 and the closely related anion exchanger AE2, which does not bind ankyrin, we have defined a 40-residue region of AE1 between positions 155 and 195 that is also essential for ankyrin binding.

Anion Exchange Protein 1, Erythrocyte↗

PubMed1996

A novel murine model for the assessment of human CD2-related reagents in vivo.

CD2 is a T cell surface glycoprotein that mediates both cell-cell adhesion and transmembrane signal transduction. To construct a model for the in vivo evaluation of human (h)CD2 function and hCD2-related reagents, hCD2 transgenic mice and murine (m)CD2 knockout mice were crossed, and the F2 generation selected for mCD2-hCD2+ animals by fluorescent flow cytometry. The mCD2-hCD2+ mice are healthy and have a normal distribution of mCD3, mCD4, and mCD8 in thymus, spleen, and lymph node. Therefore expression of the hCD2 transgene does not appear to disrupt normal T cell development. The functionality of hCD2 was demonstrated by T lymphocyte proliferation upon stimulation by combined anti-CD2 plus anti-CD2R (anti-T11(2) plus anti-T11(3)) mAbs. Anti-T11(2) plus anti-T11(3) anti-human CD2 mAbs also induced proliferation of mCD2+hCD2+ F1 lymphocytes, but not mCD2+hCD2- wild-type murine lymphocytes. Either an anti-murine or the human CD2 specific (anti-T11(1)) mAbs inhibited proliferation in alloantigen, PHA, or anti-CD3 mAb stimulated cultures and inhibited only cells bearing the appropriate cognate CD2. In vivo studies of immune function yielded results consistent with these in vitro assays. Thus, anti-T11(1) mAb suppressed contact sensitivity in vivo in the transgenic/knockout mice. mCD2-hCD2+ mice treated with anti-T11(1) or LFA-3 fusion proteins also showed significant prolongation of cardiac allograft survival. This prolongation was associated both with depletion and down-modulation of CD2 on remaining T cells. These data suggest that the transgenic/knockout mice provide a useful in vivo model for the assessment of hCD2-related reagents and CD2 function, free from any potential interactions with mCD2 and mCD2 ligands.

Animals↗

PubMed1996

Involvement of a single-stranded DNA binding protein, ssCRE-BP/Pur alpha, in morphine dependence.

We have purified a nuclear protein from mouse cerebella that binds to single-stranded oligo-DNA of cAMP response element and is modulated by morphine treatment. Isolation of the cDNA clone showed that the nuclear protein (ssCRE-BP) was identical to Pur alpha, a DNA binding protein for single-stranded purine-rich sequences that was originally isolated as a replication factor. ssCRE-BP/Pur alpha and mRNA were abundant in the brain. The levels of ssCRE-BP/Pur alpha and the transcript were not changed by chronic morphine treatment, however, the levels of an activator of ssCRE-BP/Pur alpha, which is necessary for the DNA binding, may be modulated by the treatment.

3T3 Cells↗

PubMed1996

MEKA/phosducin attenuates hydrophobicity of transducin beta gamma subunits without binding to farnesyl moiety.

Hydrophobic modifications of transducin (T) gamma, such as farnesyl-and carboxyl-methylation, are essential for the association of T beta gamma with the photoreceptor disc membrane, and MEKA/phosducin is known to inhibit the association. In this study, we examined the effect of MEKA on the hydrophobicity of T beta gamma. MEKA could bind to T beta gamma without farnesyl/carboxyl-methyl moieties as well as native T beta gamma. In the Triton X-114 phase separation assay, T beta gamma-MEKA complex was recovered in the aqueous phase, whereas T beta gamma was recover in the detergent phase. N-terminal portion of MEKA which includes T beta gamma-binding domain was not sufficient to reduce the hydrophobicity of T beta gamma or to dissociate T beta gamma from the membrane. The data suggest that MEKA attenuates the hydrophobicity of T beta gamma to result in the dissociation of T beta gamma from the membrane without directly binding to farnesyl/carboxyl-methyl moieties.

Animals↗