PubMed Health⌕ Search

Biomedical subjects

X Ren

Publications and source records attributed to X Ren.

At least 127 records · Page 7Linked to original sources
PubMed1996

[Molecular weight determination and protein analysis of GCF-5 antigens of giant cell tumor of bone].

In order to understand the tumor antigen(s) of giant cell tumor of bone (GCT), monoclonal antibody GCF-5 developed in this laboratory was employed. GCF-5 reacted specifically against the tumor cell associated antigens of GCT. The membrane proteins of tumor cell were extracted by high concentration of SDS, followed by a series of treatment processes, including SDS-PAGE, western blotting and some staining methods, such as Periodic-Schiff for glycoprotein and oil red O for lipoprotein. The molecular weight and chemical characteristics of GCF-5 antigens were analysed. The preliminary results indicated that the main GCF-5 antigens were two glycoproteinis containing few saccharide chains, one of which was 13.5KD and the other 17.4KD.

Antigens, Neoplasm↗
PubMed1996

[A study of the relevant factors of removal of common bile duct stones with endoscopic mechanical lithotripsy].

Endoscopic mechanical lithotripsy (EML) was attempted in 41 patients with common bile duct stones unable to be removed by endoscopic sphincterotomy. These patients had altogether 89 stones with stone size ranged from 5 to 30 mm (mean transverse diameter 16.6 +/- 6.0 mm and mean longitudinal diameter 20.0 +/- 7.1 mm). 31 patients had 34 stones greater than or equal to 20 mm in transverse diameter. The overall stone removal rate was 97.6%. The rate of complete removal of the stone fragments with EML at one time was 78.6% (11/14) for large, 58.8% (10/17) for middle and 0 (0/5) for small incision of the sphincter. In patients with diameter of the distal common bile duct greater than or equal to 14 mm and less than 9 mm the rate was 83.3% and 30.8% respectively and the difference was of significance (P < 0.05). The rate of complete removal for using basket and balloon catheter was 6/10 and 4/9 separately and that for combined used of these two methods 83.3% (10/12). The rate of spontaneous passage of the stone (within one week) was 1/5. It is concluded that complete removal of the stone fragments was related mainly to the length of the incision, diameter of distal common bile duct and methods of removing stone fragments after the crush with lithotripsy basket; when the stone is large and there is the no space around the stone for lithotripsy basket manipulation, a large amount of diluted contrast material is injected into the bile duct so as to increase the space and facilitate the ensonaring of the stone with lithotripsy basket. Hard stones can not be crushed easily, so they constitute one of the reasons of failure.

Adult↗
PubMed1995

Proton transfer by histidine 67 in site-directed mutants of human carbonic anhydrase III.

The ability of a histidine residue at position 67 in human carbonic anhydrase III to transfer protons in the catalytic pathway for the hydration of CO2 was investigated for a series of site-specific mutants. Wild-type carbonic anhydrase III has an arginine at this position with the C alpha of residue 67 about 9.4 A from the zinc. The active-site cavity contains no other residues capable of facile proton transfer. Rate constants for proton transfer from His 67 to the zinc-bound hydroxide were determined from the rate constants for the exchange of 18O between CO2 and water measured by mass spectrometry. A range of values for the pKa of zinc-bound water was achieved by replacement of phenylalanine with leucine and aspartate at position 198 adjacent to the zinc. Application of Marcus rate theory showed that intramolecular proton transfer involving His 67 had an intrinsic energy barrier of 1.3 +/- 0.3 kcal/mol and a thermodynamic work function for a preceding unfavorable equilibrium of 10.9 +/- 0.1 kcal/mol. We previously showed that proton transfer from histidine 64 in carbonic anhydrase III could be described by Marcus rate theory [Silverman, D. N., Tu, C. K., Chen, X., Tanhauser, S. M., Kresge, A. J., & Laipis, P. J. (1993) Biochemistry 32, 10757-10762]. In comparison, proton transfer from His 67 must overcome a more unfavorable preceding equilibrium (a larger work function) that probably represents an energy requirement for proper alignment of donor and acceptor groups plus the intervening hydrogen-bonded water. Once this alignment is achieved, the intrinsic energy barrier appears the same for His 67 or His 64.

Amino Acid Sequence↗
PubMed1995

Change of water-soluble-protein, urea-soluble-protein and membrane intrinsic protein in human senile cataract.

PURPOSE: To analyze the change of water-soluble-protein (WSP), urea-soluble-protein (USP) and membrane intrinsic protein (MIP) in human senile cataract. METHODS: The water-soluble-fractions (WSF) were prepared basically according to the method of Kibbelear, et al. But in this study, 5 mmol/L B-mercaptoethanol was added to the buffer solution. The urea-soluble-fractions (USF) were prepared basically according to the method of Kibbelear, et al. Lens fiber cell membranes were purified basically according to the method of Russell, et al. SDS-PAGE were performed according to the procedure of Laemmili, et al. using resolving gel 13% and 3% stacking gel. RESULTS: The WSP was fractionated into HM+ alpha-, beta(1-3)- and gamma-crystallin components. In nuclear cataractous lenses HM+ alpha- and B-crystallin increase, while r-crystallin decrease. The USP from clear lenses contains mainly alpha beta chains of 22KD, whereas in cataractous lenses, especially in nuclear cataractous lenses, the relative amount of the 28- and 23KD polypeptide (the components of beta-crystallin) increased markedly. Lens fiber cell MIP, clear lens and cataract lens contained the main polypeptide of 27KD (MIP) and 23KD (MP23). CONCLUSION: The water-insoluble protein, whether in quantity or in quality, plays an important role in cataract formation.

Aged↗
PubMed1994

Interactions of active-site residues and catalytic activity of human carbonic anhydrase III.

To elucidate the interactions between residues found in the active-site cavity of human carbonic anhydrase III, we have prepared a series of single and double mutants with Lys-64, Arg-67, and Phe-198 replaced with Ala, Asp, Glu, His, and Leu. Rates of catalysis were determined using 18O exchange between CO2 and water measured by mass spectrometry and initial velocity measured by stopped-flow spectrophotometry. Replacement of these residues resulted in increases in kcat/Km for CO2 hydration as much as 200-fold and increases in the pKa of the zinc-bound water by as much as 3.5 units. We conclude that the effect of replacements made at positions 64, 67, and 198 were in general additive for kcat/Km for CO2 hydration, indicating that there is no interaction between these sites that affects the catalytic interconversion of CO2 and HCO3-. One notable exception is the antagonism exhibited by the double mutant of human carbonic anhydrase III containing Glu-64 and Leu-198. The data also show that one source of the large enhancement of kcat/Km for the mutant containing Asp-198 in human carbonic anhydrase III is the presence of both Asp-198 and Lys-64; when Lys-64 was replaced with Ala, a reduction of catalytic activity was observed. These results provide an additional view of the independent interactions of amino acids that affect the catalytic pathway of isozyme III, the least active of the known carbonic anhydrase isozymes.

Binding Sites↗
PubMed1994

[The effect of medical treatment on hypertension community control--a follow-up study of more than one thousand hypertensives in East City Proper, Beijing].

Hypertension exerts a great threat on human health. Although most of countries promote the management of hypertension in communities, the role of treatment of hypertension is still controversial, it directly influences the strategy of hypertension community control. This study was based on the survey of essential and borderline hypertension in Gu-lou District of East City Proper, Beijing in 1981, and the management of 1339 hypertensives in the follow-up from 1982 to 1985, and the mortality registration in the district by Disease Surveillance System. By observing the prognosis within one year among different treatment groups (nonusers, less than 9 months and more than 9 months within one year), 1289 patients were followed up for a total of 4283 person-years. Crude analyses, stratified and logistic analyses with multi-variables adjustment of age, sex, blood pressure, with or without end-organ damage, etc. could not find that treatments had obvious effects on the reduction of incidence and mortality of stroke, mortality of CVD and all-cause in hypertensives except severe cases. The stratified analyses dy smoking also showed that smoking had a disadvantageous effects for treated hypertensives. Based on our results, we suggest that health education should be a major part of hypertension management program in community; treatments of hypertension should consider the flexible situation of patients and may not be a priority in hypertension community control program.

Aged↗
PubMed1993

Mutagenesis of structural half-cystine residues in human thioredoxin and effects on the regulation of activity by selenodiglutathione.

A human thioredoxin cDNA was modified to optimize Escherichia coli expression and subcloned into the plasmid pACA, a vector for T7 RNA polymerase-directed expression. The substitution of structural (noncatalytic) half-cystines in human thioredoxin (hTrx) was made by site-directed mutagenesis. The recombinant wild-type (wt) hTrx and its mutant C61S, C72S, and C61S/C72S were expressed and purified to homogeneity. Characterization of the wt and mutant hTrx was done with respect to redox activity with thioredoxin reductase (TR), tryptophan fluorescence, and effects of incubation with GS-Se-SG, which is believed to be the major metabolite of inorganic selenium compounds in mammalian tissues. The Km and kcat of wild-type hTrx for human placenta thioredoxin reductase (HP-TR) at pH 7.0 were 2.0 microM and 2800 min-1, respectively. The mutant proteins C61S, C72S, and C61S/C72S had Km and kcat values similar to those of the wt thioredoxin. Tryptophan fluorescence measurements showed that the wt and mutant proteins had similar stability to a denaturing agent. Incubation of fully reduced thioredoxin with 0.1 molar equivalent of GS-Se-SG resulted in continued oxidation of SH groups. After 3.5 h only 0.5 of initially 4.6 SH groups/thioredoxin remained. With the oxidized protein, a pronounced lag phase in thioredoxin reductase-dependent insulin disulfide reduction was present. Disulfide-linked dimers of the protein were present. The results clearly showed that noncatalytic cysteine residues in hTrx were oxidized accompanied by dimerization and inactivation. The activities of the mutant proteins C72S and C61S/C72S were unchanged after 3 h of incubation with GS-Se-SG. No dimer appeared of the C72S thioredoxin.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1993

In vivo animal models of body composition in aging.

We developed several techniques that provide data on body elemental composition from in vivo measurements in rats. These methods include total body potassium by whole-body counting of endogenous 40K; total body calcium (TBCa), sodium and chloride by in vivo neutron activation analysis and total body phosphorus (TBP) and nitrogen (TBN) by photon activation analysis. These elements provide information on total body fat, total body protein and skeletal mass. Measurements were made in 6-, 12- and 24-month-old rats. TBN increased slightly between 6 and 12 months but was significantly lower by 24 months, indicating a substantial loss in total body protein. Working at the National Synchrotron Light Source, we studied rat femurs by computed microtomography (CMT), and the elemental profile of the femur cortex by synchrotron-radiation induced X-ray emission (SRIXE). Although there were no significant changes in TBCa and TBP, indices of skeletal mass, CMT revealed a marked increase in the size and number of cavities in the endosteal region of the femur cortex with increasing age. The SRIXE analysis of this cortical bone revealed a parallel decrease in the endosteal Ca/P ratio. Thus, there are major alterations in bone morphology and regional elemental composition despite only modest changes in total skeletal mass.

Aging↗
PubMed1993

Alteration of crystallin polypeptides in rat lenses during the development of galactose-induced cataract.

Some striking differences in relative polypeptide abundance of crystallins were observed in normal and galactose-induced cataractous lenses of rat by means of SDS-PAGE. In the cataractous lenses a prominent band appeared at about 25 kDa and the aA chain increased markedly, whereas the relative amount of the 31 kDa band decreased substantially. These alterations are similar to the changes observed during the incubation of young mouse lenses in glucose-free medium.

Animals↗
PubMed1992

Buffer dependence of CO2 hydration catalyzed by human carbonic anhydrase I.

The steady-state kinetics of CO2 hydration catalyzed by human carbonic anhydrase I (carbonate hydro-lyase, EC 4.2.1.1) has been investigated at three pH values corresponding to different parts of the pH-rate profile. Two buffer systems with similar pKa values were used at each pH. The results show that the catalyzed rates depend on the buffer concentration but also on the chemical nature of the buffer. For example, at pH 8.8 the buffer 1,2-dimethylimidazole behaves formally as a second substrate in a 'ping-pong' mechanism yielding a maximal kcat value of 2.2 x 10(5) s-1, whereas much lower rates were obtained with Taps buffers. Similarly, at pH 7.3 1-methylimidazole yields higher rates than Mops and at pH 6.3 3,5-lutidine is more efficient than Mes. Non-Michaelis-Menten kinetics were observed with all buffers except 1,2-dimethylimidazole. In addition, while the apparent buffer activation by 1,2-dimethylimidazole can be described by a single Km value of 26 mM, the Mes concentration dependence is consistent with the presence of two components of similar magnitudes with Km values of 45 mM and 0.15 mM. These results are interpreted within the framework of the 'zinc-hydroxide' mechanism in terms of multiple pathways for the rate-contributing transfer of a proton from the zinc-bound water molecule, formed during CO2/HCO3- interconversion, to the reaction medium, thus, regenerating zinc-bound OH-.

Buffers↗
PubMed1988

Kinetics, anion binding and mechanism of Co(II)-substituted bovine muscle carbonic anhydrase.

The binding of N3- to Co(II)-substituted bovine carbonic anhydrase III was measured at various pH values by spectrophotometric titrations. The apparent Ki values were found to increase with pH in the studied range between pH 5.8 and 8.9. The inhibition of CO2 hydration by N-3 was found to be essentially uncompetitive at all investigated pH values (pH 6.3-8.9). The Ki values for the inhibition of kcat are much smaller than those obtained in the spectrophotometric titrations indicating that an enzyme form with a high affinity for N-3, presumably having a metal-bound H2O, accumulates in the steady state at saturating CO2 concentrations. Assuming that the low pH limit of Ki = 9 microM for the inhibition of kcat represents the affinity of N-3 for the Co(II)-OH2 form, a pKa value near 5 can be estimated for Co(II)-bound water from the pH dependence of N-3 binding in the absence of CO2. Measurements of time-resolved absorption spectra during CO2 hydration in the presence of a low N-3 concentration showed the transient appearance of the characteristic spectrum of the enzyme-N-3 adduct clearly demonstrating the accumulation in the steady state of an enzyme form with a high affinity for N-3. In similar experiments without inhibitor the transient formation of a spectral form corresponding to a Co(II)-OH2 species has been demonstrated. This spectral form is rather featureless lacking the absorption maxima at 618 nm and 640 nm characteristic of the Co(II)-OH- species. Our results strongly support the hypothesis that the rate-limiting step in CO2 hydration catalyzed by carbonic anhydrase III is the protolysis of metal-bound water.

Animals↗
PubMed1988

A comparison of the kinetic properties of native bovine muscle carbonic anhydrase and an activated derivative with modified thiol groups.

Steady-state and equilibrium kinetic properties of native bovine carbonic anhydrase III (carbonate hydrolyase, EC 4.2.1.1) and a derivative modified with methyl methanethiosulfonate were investigated. The modified enzyme has a markedly increased CO2 hydration activity compared to the native form with a 3-times higher value of kcat and a 6-10-times higher value of kcat/Km. Qualitatively, the activated enzyme shows the same kinetic behavior as native isoenzyme III. This is reflected in similar pH dependences of the kinetic parameters for CO2 hydration, similar solvent hydrogen isotope effects on these parameters, similar deviations from Michaelis-Menten kinetics for the HCO3- dehydration reaction, and similar behavior of the kinetics of CO2/HCO3- exchange at chemical equilibrium as measured by a 13C-NMR magnetization transfer technique. It is concluded that the conversion of -SH groups to -S-S-CH3 moieties does not change the catalytic mechanism, but leads to an increased rate of CO2/HCO3- interconversion as well as to an increased rate of proton transfer between the active site and the reaction medium.

Alkylation↗
PubMedDate not supplied

Beta-adrenergic receptor subtypes in stress-induced behavioral depression.

The purpose of this study was to examine the role of beta-adrenergic receptors in an animal model of stress-induced behavioral depression. beta-Adrenergic receptors in several brain regions and leukocytes of rats were determined by receptor binding techniques using 125I-cyanopindolol (cyp) as ligand and propranolol as displacer for total beta-adrenergic receptors, and ICI 86,406 for beta 1- and ICI 118,551 for beta 2-adrenergic receptors. We observed that the maximum number of binding sites (Bmax) and the apparent dissociation constant (Kd) of 125I-cyp binding to total beta-adrenergic receptors were increased in hippocampus of stressed rats with escape deficits (48 h after training) as compared to control rats. This increase was due to an increase in Bmax and Kd of 125I-cyp binding to beta 1-adrenergic receptors but not to beta 2-adrenergic receptors. There was no significant difference in beta 1-adrenergic receptors in cortex and cerebellum or beta 2-adrenergic receptors in hippocampus, cortex, cerebellum, or leukocytes of stressed (48 h after training) rats with escape deficits as compared to control rats. Interestingly, it was observed that beta 1- and beta 2-adrenergic receptors in various brain regions (cortex, cerebellum, and hippocampus) and beta 2-adrenergic receptors in leukocytes of stressed rats (10 days after training) were not significantly different from control rats, although escape deficits were still present. These results suggest that abnormalities in adrenergic neurotransmission are associated with an upregulation of beta 1-adrenergic receptors, which in turn may be involved in the early stages of behavioral deficits caused by uncontrollable shock.

Animals↗
PubMedDate not supplied

A novel selenocystine-beta-cyclodextrin conjugate that acts as a glutathione peroxidase mimic.

A novel artificial glutathione peroxidase mimic consisting of a selenocystine-di-beta-cyclodextrin conjugate (selenium-bridged-6, 6'-amino-selenocystine-6,6'-deoxy-di-beta-cyclodextrin), in which selenocystine is bound to the primary side of beta-cyclodextrin through the two amino nitrogen groups of selenocystine, was synthesized. The glutathione peroxidase activities of the mimic-catalyzed reduction of H(2)O(2), tert-butylhydroperoxide, and cumene hydroperoxide by glutathione are 4.1, 2.11, and 5.82 units/micromol, respectively. The first activity was 82 and 4.2 times as much as that of selenocysteine and ebselen, respectively. Studies on the effect of substrate binding on the glutathione peroxidase activity suggest that it is important to consider substrate binding in designing glutathione peroxidase mimics. The detailed steady-state kinetic studies showed that the mimic-catalyzed reduction of H(2)O(2) by glutathione followed a ping-pong mechanism, which was similar to that of the native glutathione peroxidase.

Binding Sites↗