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Biomedical subjects

D Xie

Publications and source records attributed to D Xie.

At least 73 records · Page 4Linked to original sources
PubMed1999

[Morphological observations of the medial olivocochlear efferents in gerbils with chronic gentamicin ototoxicity].

OBJECTIVE: To determine the possible involvement of the medial olivocochlear efferents (MOC) in chronic aminoglycoside ototoxicity, the morphological changes of the MOC efferents in the cochlea of Mongolian gerbils and their correlations with the impairment of the outer hair cells (OHCs) were observed. METHODS: The animals were given gentamicin in a dose of 150 mg/kg a day for 10 days. The distributions and densities of the MOC efferents were examined using the modified histochemical staining for acetylcholinesterase (AChE) and the numbers of OHCs were numerated with toluidine blue and Ehrlich haematoxylin staining on the surface preparation. RESULTS: Significant damage to the MOC fibers and terminals as well as OHCs was noted in the basal turn of the cochlea, especially in those animals examined in 7th and 11th week after the gentamicin administration. The site of the greatest impairment of OHCs was consistent with that of the MOC efferents. CONCLUSION: The results suggested that damage to the MOC efferents might play an important role in gentamicin-induced impairment of OHCs.

Animals↗
PubMed1999

[Study on the human serums by absorption spectra].

The absorption spectra of normal human and cancer patient serums were measured, and those of human serum after adding some cancer cells were also measured. The results showed some difference in absorbance among the three kinds of serums. Some useful results were obtained.

Humans↗
PubMed1998

Functional characterization of the protease of human endogenous retrovirus, K10: can it complement HIV-1 protease?

To investigate the biochemical properties of the protease encoded by the human endogenous retrovirus, K10 (HERV-K), 213 amino acids of the 3'-end of the HERV-K protease (PR) open reading frame were expressed in Escherichia coli. Autocatalytic cleavage of the expressed polypeptide resulted in an 18.2 kDa protein which was shown to be proteolytically active against a fluorogenic peptide used as a substrate for HIV-1 protease. On the basis of sequence homology and molecular modeling, the 106 N-terminal amino acids of HERV-K PR were predicted to comprise a retroviral protease core domain. An 11.6 kDa protein corresponding to this region was expressed and shown to be a fully functional enzyme. The 11.6 kDa domain of HERV-K PR is unusually stable over a wide pH range, exhibits optimal catalytic activity between pH 4.0 and 5.0, and exists as a dimer at pH 7.0 with a Kd of 50 microM. Like HIV-1 PR, the HERV-K PR core domain is activated by high salt concentrations and processes HIV-1 matrix-capsid polyprotein at the authentic HIV-1 PR recognition site. However, both the 18.2 and 11.6 kDa forms of HERV-K PR were highly resistant to a number of clinically useful HIV-1 PR inhibitors, including ritonavir, indinavir, and saquinavir. This raises the possibility that HERV-K PR may complement HIV-1 PR during infection, and could have implications for protease inhibitor therapy and drug resistance.

Amino Acid Sequence↗
PubMed1998

Thermodynamics of a transition state analogue inhibitor binding to Escherichia coli chorismate mutase: probing the charge state of an active site residue and its role in inhibitor binding and catalysis.

Electrostatic interactions play important roles in the catalysis of chorismate to prephenate by chorismate mutase. Mutation of Gln88 to glutamate in the monofunctional chorismate mutase from Escherichia coli results in an enzyme with a pH profile of activity significantly different from that of the wild type protein. To investigate whether the mutation alters the substrate binding process or the catalysis, we have directly determined the thermodynamic parameters of a transition state analogue inhibitor binding to the wild-type chorismate mutase and its Q88E mutant using isothermal titration calorimetry. The results demonstrate that solvent reorganization and hydrophobic interactions contribute the predominant free energy to inhibitor binding. The charge state of Glu88 in the Q88E mutant was experimentally determined and was shown to be protonated at pH 4.5 and ionized at pH 7.8, consistent with earlier hypotheses. Most surprisingly, inhibitor binding energetics do not exhibit significant pH dependency for both enzymes. Our findings indicate that the charge state of Glu88 has a small impact on inhibitor binding but plays an important role in the catalytic process.

Amino Acid Substitution↗
PubMed1998

Lidocaine prolongs the safe duration of circulatory arrest during deep hypothermia in dogs.

PURPOSE: To test the hypothesis that lidocaine prolongs the safe period of circulatory arrest during deep hypothermia. METHODS: Sixteen dogs were subjected to cooling, first surface cooling to 30 degrees C and then core cooling to 20 degrees C rectal temperature). The circulation was then stopped for 90 min. In the lidocaine group, 4 mg.kg-1 lidocaine was injected into the oxygenator two minutes before circulatory arrest and 2 mg.kg-1 at the beginning of reperfusion and rewarming. The control group received equivalent volumes of normal saline. Post-operatively, using a neurological deficit scoring system (maximum deficit score-100; minimum-zero indicating that no scored deficit could be detected). Neurological function was evaluated hourly for six hours and then daily for one week, the pharmacokinetic parameters were calculated using one compartment model. RESULTS: On the seventh day, the neurological deficit score and overall performance were better in the lidocaine (0.83 +/- 2.04) than in the control group (8.33 +/- 4.08 P < 0.05). During the experiment, the base excess values were also better in the lidocaine than in the control group (at 30 min reperfusion: -4.24 +/- 1.30 vs -8.20 +/- 2.82 P < 0.01, at 60 min reperfusion was -3.34 +/- 1.87 vs -7.52 +/- 2.40 (P < 0.01). On the eighth day the extent of pathological changes were milder in the lidocaine group than that in the control group. The elimination half life of lidocaine was 40.44 +/- 7.99 during hypothermia and 2.01 +/- 4.56 during rewarming. CONCLUSIONS: In dogs lidocaine prolongs the safe duration of circulatory arrest during hypothermia.

Anesthetics, Local↗
PubMed1998

[Study on the effects of hyaluronic acid-streptomycin perfusion through the round window on the function and morphology in guinea pig inner ears].

To investigate the effects of the hyaluronic acid-streptomycin (HA-SM) perfusion through round window on the function and morphology of the inner ear in guinea pig, membrous labyrinth mapping, temporal bone section after celloidin embedding, transmission electron microscopy, electrocochlegraphy (ECochG) and electronystagmography (ENG) were examined. The nystagmus duration induced by caloric test was obviously reduced in comparison with that of the preoperation (P< 0.01), while the action potential(AP) by ECochG was not obviously changed. The sensory cells of estibular organs were severely damaged, while the morphology of corti's organs were significantly damaged after HA-SM perfusion. The results suggest that the HA-SM perfusion through the round window may selectively destroy the vestibular function, whereas the auditory function is not obviously damaged.

Animals↗
PubMed1998

[Study on the relationship between deoxyribonucleoside triphosphate (dNTP) pools and cell transformation].

Deoxyribonucleoside triphosphate (dNTP) pools were measured in normal BALB/c3T3 cells, transformation-treated cells and transformed cells with reverse-phase HPLC. The fluctuation of dNTP pools was similar after cells were treated with alkylating mutagens glycidyl methacrylate (GMA) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). The gap between (dGTP + dATP) pools and (dTTP + dCTP) pools was greatly intensified. The measurements also indicated that the dNTP pools in transformed cells were quite different from those in normal cells. The results suggest that dNTP pools may play an important role in cell transformation.

3T3 Cells↗
PubMed1998

[Diagnosis and treatment of Ramsay Hunt syndrome (a report of 39 cases)].

Thirty-nine cases with Ramsay Hunt syndrome were presented, in which 23 cases were firmly diagnosed early, and others were misdiagnosed to be Bell's palsy in 9 cases, sudden sensorineural hearing loss in 5, herpes zoster pharyngitis in 1 and acute suppurative otitis media in 1, respectively. All patients were treated with prednisone or dexamethasone for 3 weeks. The results of treatment were as follows: complete recovery in 27 cases, residual facial paralysis in 12 patients, in which 11 had sensorineural hearing loss. We concluded that: 1. When patients present idiopathic facial paralysis associated with objective sensorineural hearing loss, Hunt syndrome should be suspected even in the absence of vesicles. 2. Treatment with steroid and antiviral agent is needed. There is no significant difference in the effects between oral and intravenous steroid therapy. 3. The poor prognosis may relate with severe facial paralysis accompanying severe hearing loss. 4. Acoustic stapedius reflex in patients with mild or no hearing loss is useful for defining the involved sites and evaluating the prognosis.

Adolescent↗
PubMed1998

[Immobilization of N-acetyl-1,6-diaminohexane(NADAH) on diol-silica and study of mechanism of NADAH and hexamethylene bisacetamide(HMBA) inducing tumor cells differentiation].

A method for separating NADAH and its immobilization onto diol-silica have been developed. HMBA and its meabolite NADAH were used as the inducers for many types of tumor cells' differentiation. The inducing mechanism of HMBA is not clear yet. Experiments show that HMBA and NADAH have a relatively strong hydrophobic reaction and hardly ion-exchange with some DNA and some proteins of the cytosolic fraction of HL-60 human acute progranulocytic leukemia cells, BIU-87 human bladder carcinoma cells and the human erythrocyte membrane, when utilizing immobilization of NADAH. The retention time of the proteins and DNA was longer than that of the phosphatides. These results show that the adsorption of HMBA and NADAH with some proteins and DNA is higher than that with the phosphatides. The expected receptor binding biospecifically with HMBA has not been found.

Acetamides↗
PubMed1998

[An analysis of hepatitis G virus infection status in various cases of hepatitis in Guangdong area].

To understand hepatitis G virus(HGV) infection status in different clinical types of hepatitis, 721 cases with hepatitis were analyzed in this article. PT-PCR technique was applied to amplify HGV gene, and heat denaturation method was used to extract HCV-RNA from sera. After turning to cDNA by reverse transcription(PT), a nested amplification was conducted and a 238 bp specific fragment was obtained. In this way, serum specimens of 721 cases with hepatitis, were tested including 587 hospitalized cases. 80 cases were found to be HGV RNA positive, of which positive rate of patients with non A-E hepatitis was high, HCV and HBV + HCV came the second, chronic severe hepatitis and liver cirrhosis (LC) was the lower. Results of research showed that HGV infection is prevalent to a certain degree among patients with viral hepatitis in Guangdong area.

Flaviviridae Infections↗
PubMed1998

[Purification and properties of antifungal protein X98III from Bacillus subtilis].

Bacillus subtilis BS-98 is one of the antagonistic strains strongly against plant fungal pathogens such as Physalospora piricola Nose. The antifungal protein was purified by ammonium sulphate precipitation and column chromatography on Sephadex G-100 and DEAE-cellulose, and it was named X98III. Molecular weight of X98III is 59,000 by SDS-PAGE and PI value is 4.50 by PAG-IEF, respectively. X98III was demonstrated as glycoprotein and lipoprotein by CAM (cellulose acetate membrane) electrophoresis and special staining. We estimated it contains 6% saccharides by using DNS methods. This protein was also found to be thermostabale and partially sensitive to proteinases. The amino acid analysis of the protein X98III showed that it comprises of 11 different amino acids and Glu, Tyr, Cys are the abundant amino acids. No Asp, Phe and Met were found. Purified X98III has strong inhibiting activity against the pathogens of Physalospora piricola, Phoma asparagi, etc. The antifungal mechanism of X98III was mainly disintegration of the cell wall to make the hyphae abnormal and the spores germinate abnormally or can not germinate at all.

Amino Acids↗
PubMed1998

[Spectroscopic characterization of a new terbium complex with red fluorescence].

Usually, the emission spectra of terbium complexes exhibit 490 nm, 545 nm bands (green color) under the excitation of UV light, which is ascribed to the two 5D4 - 7F6, 5D4 - 7F5 transition. In this report, pink red fluorescence under the excitation of 300-370nm UV light is observed for a new 1, 10-phenanthroline-2-naphthoate complex of terbium. The fluorescence, FT-IR, ICP and X-ray fluorescence analysis were performed. The results showed that under the perturbation of the ligands field, the probability of 5D4 - 7F3 transition of terbium ion was greatly enhanced, while the probability of 5D4 - 7F5 transition was greatly decreased in the complex. Therefore, pink-red fluorescence was observed for the title complex.

English Abstract↗
PubMed1998

[Investigation on the luminescence properties of Er ions in Er-doped sol-gel silica glasses].

Er-doped sol-gel silica glasses which have room temperature photoluminescence at 1.54 microm and doped concentration of 10(20)/cm(3) have been prepared. The examination results Indicated: (1) PL spectrum has largest intensity at the 0.5W% doped concentration; (2) the intensity of PL decreased 74%, when the temperature raised form 4K to 300K; (3) The Er ion and O ions formed complexes with coordinated numbers of 8 or 9.

English Abstract↗
PubMed1997

Dissection of the pH dependence of inhibitor binding energetics for an aspartic protease: direct measurement of the protonation states of the catalytic aspartic acid residues.

The catalytic activity and inhibitor binding energetics of enzymes are often pH-dependent properties. Aspartic proteases comprise an important class of enzyme targets for structure-based drug design. We have performed a complete thermodynamic study of pepstatin binding to plasmepsin II, an aspartic proteinase found in Plasmodium falciparum, using isothermal titration calorimetry and circular dichroism. Thermodynamic parameters (DeltaG, DeltaH, DeltaCp, and DeltaS) were measured as functions of both pH and temperature. In the pH range from 4.5 to 7.0, pepstatin binding is accompanied by proton transfer between the solvent and the complex. We used thermodynamic proton linkage theory to derive both the pH-independent binding energetics for pepstatin and the number and pKa values of ionizable residues whose pKa values change during ligand binding. These residues were identified as the two catalytic aspartates, with pKas of 6.5 and 3.0, and His 164, with a pKa of 7.5, based on the three-dimensional structure of the pepstatin-plasmepsin II complex. At pH 5.0, where the protease has optimum activity, the proton transfer process contributes almost 40% of the total binding free energy change and the total charge of the active-site aspartic acid residues is -1. These experimental results provide direct measurement for the protonation states of the catalytic aspartates in the presence of bound ligands. Comparison of the thermodynamic and structural data for pepstatin binding with human cathepsin D, a lysosomal aspartic protease that shares 35% sequence identity with plasmepsin II, suggests that the energetic differences between these two proteins are due to a higher interdomain flexibility in plasmepsin II.

Animals↗
PubMed1997

Phosphorylation of serine 392 stabilizes the tetramer formation of tumor suppressor protein p53.

Tumor suppressor protein p53 is a tetrameric phosphoprotein that activates transcription from several cell cycle regulating genes in response to DNA damage. Tetramer formation is critical to p53's ability to activate transcription; however, posttranslational modifications and protein stabilization also contribute to p53's ability to activate transcription. To determine if phosphorylation affects tetramer formation, we synthesized phosphopeptides corresponding to residues 303-393 of human p53, which includes the domain responsible for tetramer formation. Phosphate was chemically incorporated at Ser315, Ser378, or Ser392 and also at both Ser315 and Ser392. Equilibrium ultracentrifugal analyses showed that phosphorylation at Ser392 increased the association constant for reversible tetramer formation nearly 10-fold. Phosphorylation of either Ser315 or Ser378 had little effect on tetramer formation, but phosphorylation of Ser315 largely reversed the effect of phosphorylation at Ser392. Analyses by calorimetry demonstrated that phosphorylation may influence subunit affinity (and, in turn, DNA binding) by an enthalpy-driven process, possibly between the C-terminal residues and the region immediately adjacent to Ser315. The Kd for the tetramer-monomer transition of the unphosphorylated p53 C-terminal domain was determined to be approximately 1-10 microM. Thus, in normal, undamaged cells p53 may be largely monomeric. Enhancement of tetramer formation through phosphorylation of Ser392, coupled with a DNA-damage-induced increase in its nuclear concentration, could provide a switch that activates p53 as a transcription factor in response to DNA damage.

Biopolymers↗
PubMed1997

Hydrophobic chromatography of the HL-60 cellular fraction co-binding with hexamethylene bisacetamide.

Methods of separating N-acetyl-1,6-diaminohexane (NADAH) and its immobilization to diol-silica have been developed. Hexamethylene bisacetamide (HMBA) and its metabolite NADAH are used as inducers of leukemia cell differentiation. The inducing mechanism of HMBA is still not clear. Experiments show that HMBA and NADAH undergo relatively strong hydrophobic reactions and do not readily undergo ion-exchange with the proteins of the cytosolic fraction of HL-60 cells during immobilization of NADAH; the retention time of the proteins was longer than that of the phosphatides. These results show that the adsorption of HMBA and NADAH to proteins was higher than that to phosphatides. The expected biospecific receptor binding with HMBA has not been found.

Acetamides↗
PubMed1997

Effect of phosphorylation on tetramerization of the tumor suppressor protein p53.

Human tumor suppressor protein p53 is a 393-amino acid phosphoprotein that enhances transcription in response to DNA damage from several genes that regulate cell cycle progression. The tetrameric state of p53 is critical to wild-type function; the p53 tetramerization element is located in the C-terminal region of the protein. This region is phosphorylated at several evolutionarily conserved serines, suggesting that phosphorylation may be an important regulator of p53 function. In order to determine the effect of phosphorylation on tetramer formation, we synthesized phosphopeptides corresponding to p53(Ser303-Asp393) with phosphate incorporated at Ser315, Ser378, or Ser392, and at both Ser315 and Ser392. Equilibrium ultracentrifugation analysis showed that phosphorylation at Ser392 increased the association constant for tetramer formation nearly ten-fold. By itself, phosphorylation at Ser315 or Ser378 had little effect on tetramer formation, but Ser315 largely reversed the effect of phosphorylation at Ser392. Analysis by calorimetry suggests that phosphorylation may influence subunit affinity by an enthalpy driven process.

Amino Acid Sequence↗