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Biomedical subjects

Z He

Publications and source records attributed to Z He.

At least 199 records · Page 11Linked to original sources
PubMed1996

Purification and characterization of an oxygen-sensitive, reversible 3,4-dihydroxybenzoate decarboxylase from Clostridium hydroxybenzoicum.

A 3,4-dihydroxybenzoate decarboxylase (EC 4.1.1.63) from Clostridium hydroxybenzoicum JW/Z-1T was purified and partially characterized. The estimated molecular mass of the enzyme was 270 kDa. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis gave a single band of 57 kDa, suggesting that the enzyme consists of five identical subunits. The temperature and pH optima were 50 degrees C and pH 7.0, respectively. The Arrhenius energy for decarboxylation of 3,4-dihydroxybenzoate was 32.5 kJ . mol(-1) for the temperature range from 22 to 50 degrees C. The Km and kcat for 3,4-dihydroxybenzoate were 0.6 mM and 5.4 x 10(3) min(-1), respectively, at pH 7.0 and 25 degrees C. The enzyme optimally catalyzed the reverse reaction, that is, the carboxylation of catechol to 3,4-dihydroxybenzoate, at pH 7.0. The enzyme did not decarboxylate 2-hydroxybenzoate, 3-hydroxybenzoate, 4-hydroxybenzoate, 2,3-dihydroxybenzoate, 2,4-dihydroxybenzoate, 2,5-dihydroxybenzoate, 2,3,4-trihydroxybenzoate, 3,4,5-trihydroxybenzoate, 3-F-4-hydroxybenzoate, or vanillate. The decarboxylase activity was inhibited by 25 and 20%, respectively, by 2,3,4- and 3,4,5-trihydroxybenzoate. Thiamine PPi and pyridoxal 5'-phosphate did not stimulate and hydroxylamine and sodium borohydride did not inhibit the enzyme activity, indicating that the 3,4-dihydroxybenzoate decarboxylase is not a thiamine PPi-, pyridoxal 5'-phosphate-, or pyruvoyl-dependent enzyme.

Amino Acid Sequence↗
PubMed1996

Synchrony in binary-oscillator networks with local couplings.

Since synchronous oscillations in the visual cortex may be responsible for some features of visual scene, many works have been done to study the dynamics of oscillatory neural networks. Most of the oscillator network models rely on a global connection to reach synchronization. Here we propose a class of neural network models based on a simplified binary-oscillator, and find that the neurons of networks with local couplings will be in global synchrony under some criteria.

Models, Neurological↗
PubMed1996

[In vitro transformation of human embryonic nasopharyngeal epithelial cells with Epstein-Barr virus].

UNLABELLED: EBV from B95-8 cells were used to infect directly the human embryonic nasopharyngeal epithelial (HENE) cells in vitro. RESULTS: Primary HENE cells treated with EBV did not have a significantly increased colony-forming rate in soft agarose, while cells that were treated with EBV in combination with tumor promotor 12-O-tetradecanoylphorbol-13 acetate (TPA) showed a marked increase in colonyforming rate from 0-5 cells to 20-40 colonies per 10,000 cells. (2) HENE cells treated with inactivated (56 degrees C for 30 minutes) virus, in spite of adding TPA at the same time, did not significantly increase agarose colony forming rate; (3) HENE cells treated with TPA after EBV exposure for a week had significantly increased colony forming rate in soft agarose, while the cells exposed to EBV after using TPA treatment did not. In addition, EBV BNLF1 (LMP) fragment in HENE cells treated with EBV or EBV+TPA was detected by polymerase chain reaction using specific oligonucleotide primer. Only HENE cells treated with EBV+TPA presented positive band in aragose gel electrophoresis. The results suggest that transformation of EBV on HENE cells depends on infectious virus and an intact viral genome. TPA not only can promote the transformation, but also promote EBV's entering HENE cells.

Base Sequence↗
PubMed1995

L-arginine ameliorates cerebral blood flow and metabolism and decreases infarct volume in rats with cerebral ischemia.

Effects of L-arginine, 300 mg/kg, i.p., on the regional cerebral blood flow (rCBF), brain metabolism, and infarct volume were examined in spontaneously hypertensive rats subjected to occlusion of both left middle cerebral artery and left common carotid artery. Rats treated with L-arginine had higher rCBF, determined by hydrogen clearance method, in the ischemic core (7 +/- 1 ml/100 g/min, mean +/- S.E.M.) and penumbral regions (16 +/- 2) than did rats treated with saline (5 +/- 0 and 7 +/- 1, respectively). Simultaneously, L-arginine attenuated metabolic derangement in the ischemic tissue at 60 min, i.e. well maintained adenosine triphosphate (ATP) in ischemic region (1.29 +/- 0.07 mmol/kg in L-arginine group vs. 1.05 +/- 0.06 in saline group), and also close to normal levels in ATP (2.61 +/- 0.02 mmol/kg vs. 2.45 +/- 0.05), glucose (2.29 +/- 0.12 mmol/kg vs. 1.80 +/- 0.17) and lactate (1.63 +/- 0.10 mmol/kg vs. 2.24 +/- 0.21) in periischemic region. In another experiment, the effects of L-arginine on rCBF in the subcortical regions and on infarct volume were evaluated. L-arginine, compared with saline, increased rCBF by 8 ml/100 g/min in the ischemic side and reduced infarct volume by 29% at 24 h of ischemia. These findings support that L-arginine may be potentially useful for the treatment of acute cerebral ischemia.

Animals↗
PubMed1995

Mechanism-based inactivation of porcine kidney diamine oxidase by 1,4-diamino-2-butene.

Cis- and trans-1,4-diamino-2-butene are substrates and potent inactivators of porcine kidney diamine oxidase. Evidence from absorption and NMR spectra indicates that both are oxidized to pyrrole. Both substrates are irreversible mechanism-based inactivators of the enzyme, although the trans isomer is more potent and results in complete inactivation in a reaction which follows pseudo-first-order kinetics with an apparent Ki of 0.34 mM and a second-order inactivation constant of 500 M-1 s-1. Under the same conditions, 46% of the activity remains when the enzyme is reacted with cis-1,4-diamino-2-butene. Trans-4-amino-2-butenal, the product of oxidation of the trans diamine, has been synthesized and shown to undergo cyclization to pyrrole in a concentration-dependent manner, approaching second-order at low concentrations. Trans-4-amino-2-butenal is itself a potent irreversible inhibitor with IC50 of 2.5 microM. We propose that the irreversible inactivation by both cis- and trans-1,4-diamino-2-butene involves attack by a protein-based nucleophilic residue on the unsaturated aminoenal products of the enzymatic reactions, resulting in a covalent adduct. Cyclization of the cis-aminoenal to pyrrole is much more rapid than in the trans case, thus it is less available for inhibitory reaction with the protein.

Aldehydes↗
PubMed1995

Precise localization of aphidicolin-induced breakpoints on the short arm of human chromosome 3.

The common fragile site at 3p14.2 (FRA3B) has been described as the most active fragile site in the human genome. This locus may predispose chromosome 3 to specific losses due to deletions and translocations that have been associated with several malignancies, including hereditary renal cell carcinoma. We have previously described induction of breakage around FRA3B using aphidicolin in a somatic cell hybrid whose only human component was a single intact chromosome 3. That work led to the isolation of hybrids with breakpoints in the 3p13-p21.1 region with loss of all sequences distal to their respective breakpoints. In this report we describe the further characterization of the breakpoints in many of these cell lines using newly available molecular markers. We also report the identification of YAC clones that span the breakpoints present in many of these hybrids.

Aphidicolin↗
PubMed1995

Cyanide inhibition of porcine kidney diamine oxidase and bovine plasma amine oxidase: evidence for multiple interaction sites.

The interactions of cyanide and phenylhydrazine with porcine kidney diamine oxidase (PKDAO) and bovine plasma amine oxidase (BPAO) (EC 1.4.3.6) have been investigated. Cyanide displays mixed noncompetitive inhibition against amine substrates and also against O2. EPR spectroscopy shows that cyanide binds to an equatorial site on Cu(II) and can be displaced by chloride, which is not an inhibitor, without recovery of activity, indicating that Cu(II)-bound cyanide is not inhibitory. 14CN-. studies have shown that one cyanide in PKDAO and two in BPAO are covalently and irreversibly bound per enzyme dimer at an unknown site, even under conditions where cyanide is not bound to Cu(II). These cyanides have no effect on activity or on binding of phenylhydrazine to the enzymes. Cyanide also binds reversibly to the organic cofactor in both enzymes, presumably as a cyanohydrin, leading to the observed mixed noncompetitive inhibition against substrate. In both enzymes, two phenylhydrazines react per enzyme dimer. The kinetics of phenylhydrazine titration are affected by cyanide, which indicates that phenylhydrazine and cyanide react at the same carbonyl group in the enzymes. The results suggest that inhibition of amine oxidases by cyanide is through a carbonyl reagent and a Cu(I) ligand rather than through a Cu(II) ligand.

Amine Oxidase (Copper-Containing)↗
PubMed1995

RPA involvement in the damage-recognition and incision steps of nucleotide excision repair.

Human replication protein (RPA) functions in DNA replication, homologous recombination and nucleotide excision repair. This multisubunit single-stranded DNA-binding protein may be required to make unique protein-protein contacts because heterologous single-stranded binding proteins cannot substitute for RPA in these diverse DNA transactions. We report here that, by using affinity chromatography and immunoprecipitation, we found that human RPA bound specifically and directly to two excision repair proteins, the xeroderma pigmentosum damage-recognition protein XPA (refs 8, 9) and the endonuclease XPG (refs 10-13). Although it had been suggested that RPA might function before the DNA synthesis repair stage, our finding that a complex of RPA and XPA showed a striking cooperativity in binding to DNA lesions indicates that RPA may function at the very earliest stage of excision repair. In addition, by binding XPG, RPA may target this endonuclease to damaged DNA.

Chromatography, Affinity↗
PubMed1995

Purification and characterization of an oxygen-sensitive reversible 4-hydroxybenzoate decarboxylase from Clostridium hydroxybenzoicum.

A 4-hydroxybenzoate decarboxylase from the anaerobe Clostridium hydroxybenzoicum strain JW/Z-1T was purified and partially characterized. It had an apparent molecular mass of 350 kDa and consisted of six identical subunits of 57 kDa each. The temperature optimum for the decarboxylation was approximately 50 degrees C, the optimum pH 5.6-6.2. The pI of the enzyme was 5.1. The activation energy for decarboxylation of 4-hydroxybenzoate was 65 kJ.mol-1 (20-37 degrees C). The enzyme also catalyzed decarboxylation of 3,4-dihydroxybenzoate. The apparent Km and kcat values obtained for 4-hydroxybenzoate were 0.40 mM and 3.3 x 10(3) min-1, and for 3,4-dihydroxybenzoate 1.2 mM and 1.1 x 10(3) min-1, respectively, at pH 6.0 and 25 degrees C. The enzyme activity was not influenced by the addition of biotin or avidin to either the crude cell extracts or the purified enzyme. The p-hydroxyl group of hydroxybenzoate appears to be essential for binding by the enzyme. The N-terminal amino acid sequence shows some similarity to the uroporphyrinogen decarboxylases from Synechococcus and Saccharomyces. The enzyme catalyzed the reverse reactions, that is, the carboxylation of phenol to 4-hydroxybenzoate and of catechol to 3,4-dihydroxybenzoate. The carboxylation did not require ATP.

Amino Acid Sequence↗
PubMed1995

Changes of gastric acid secretion and somatostatin after Roux-en-Y cholangiojejunostomy.

The authors studied the changes of gastric acid secretion and determined the levels of somatostatin (SS) and gastrin (Gn) in blood, gastric juice and pyloric antrum in 28 patients Roux-en-Y with cholangiojejunostomy (CJR-Y). Gastric acid hypersecretion was identified, and the level of SS was found dropped while the content of Gn increased in these specimens. It was considered that the reduction of SS synthesis and secretion might have a key effect on the mechanism of secondary gastric acid hypersecretion in patients after CJR-Y.

Adult↗
PubMed1995

[Anti-metastasis gene nm23 expression in buccal cancer studied by immunohistochemistry stain].

The nm23 gene has been proposed as a candidate metastasis-suppressor gene. In this study we used immunohistochemistry stain for polyclone antibody nm23 in 62 cases of buccal carcinoma. The results showed that both the epithelium of mucosa and the duct epithelium of minor salivary gland nearby the tumor were strong positive. The stains on tumor cells were various. 31.0% (9/29) of the cases with lymph-node metastasis were positive while 57.6% (19/33) of those with no lymph-node metastasis positive (P = 0.044). Histologically, 59.4% (19/32) of the cases were positive in Grade I, while 37.5% (6/16) positive in Grade II and 21.4% (3/14) in Grade III (P = 0.048). The positive rate increased with the patients survival time from less than 3 years to 3-5 years and more than 5 years correspondingly, but there was no statistical significance (P = 0.126). The results suggested that the expression of nm23 might be one of the factors influencing the lymph-node metastasis and prognosis of buccal cancer.

Carcinoma, Squamous Cell↗
PubMed1995

[Use of perfluorodecalin in giant retinal tear retinal detachment surgery].

Perfluorodecalin is used for the treatment of 11 eyes with giant retinal tear retinal detachment surgery. Perfluorocarbon liquids can be used as an operative hydrodynamic tool during vitreous surgery. The high specific gravity of perfluorocarbon liquids allows an easy unfolding of the inverted flap and flattening of the retina, displacing the subretinal fluid and stabilizing the retina for membrane peeling. Endophotocoagulation may be performed under optimal visual condition. The success rate was 81.8% (9/11). The retina failed to reattach in patients with advanced proliferative vitreoretinopathy. The surgical techniques were discussed.

Adolescent↗
PubMed1994

The structure of the human sterol carrier protein X/sterol carrier protein 2 gene (SCP2).

Sterol carrier protein X (SCPx) is a 58-kDa protein that is localized to peroxisomes. The amino acid sequence of the protein suggests that SCPx may function as a thiolase. The gene encoding SCPx also codes for a 15.3-kDa protein called sterol carrier protein 2 (SCP2). Here we report the structure of this gene (SCP2), which spans approximately 80 kb and consists of 16 exons and 15 introns. Multiple transcription start sites were identified. The 5' flanking region has characteristics of other peroxisomal protein promoters, which include the absence of a TATA box and a G+C-enriched region containing several reverse GC boxes.

Acetyl-CoA C-Acetyltransferase↗
PubMed1994

DETERMINATION OF PROTEIN SYNTHESIS IN RAINBOW TROUT, ONCORHYNCHUS MYKISS, USING A STABLE ISOTOPE

It has been suggested (Houlihan, 1991) that the consumption of 1 g of protein in a variety of species of fish stimulates the synthesis of, approximately, an equal amount of protein. Although synthesis of protein may account for as much as 40 % of the whole-animal oxygen consumption (Lyndon et al. 1992), only about 30 % of the synthesized proteins are retained as growth (Houlihan et al. 1988; Carter et al. 1993a,b). Thus, one focus of attention is the potential advantage gained by fish in allocating a considerable proportion of assimilated energy to protein turnover in contrast to relatively low-cost, low-turnover protein growth (Houlihan et al. 1993). Rates of protein synthesis in several species of fish have been measured using radioactively labelled amino acids, frequently given as a flooding dose (reviewed by Fauconneau, 1985; Houlihan, 1991). These measurements cannot be made for longer than a few hours because of the decline in specific radioactivity in the amino acid free pool. However, as protein synthesis rates vary during the course of a day as a result of the post-prandial stimulation, and since radiolabelled amino acid methodology is invasive, short-term and terminal, it has been difficult to be certain of the relationship between protein growth measured in the long term and protein synthesis rates measured in the short term. This paper addresses these problems by developing a method using 15N in orally administered protein to measure protein synthesis rates in fish over relatively long periods, the aim being to use procedures that are as non-invasive and repeatable as possible. The use of stable isotopes to measure protein metabolism is well established in terrestrial mammals (see Rennie et al. 1991; Wolfe, 1992), but to our knowledge the only published data for aquatic ectotherms are on the blue mussel (Mytilus edulis L.) (Hawkins, 1985). In the present study, rates of protein synthesis of individual rainbow trout [Oncorhynchus mykiss (Walbaum)] were calculated from the enrichment of excreted ammonia with 15N over the 48 h following the feeding of a single meal (dose) containing protein uniformly labelled with 15N by use of an end-point stochastic model (Waterlow et al. 1978; Wolfe, 1992). Application of this type of modelling would appear to be ideal for measuring ammonotelic fish nitrogen metabolism since, unlike the situation in mammals, the catabolic flux of amino acids through urea is very small. Further, ammonia is excreted directly into the surrounding water via the gills and is not stored for any length of time, in contrast to the situation in mammals, so the rate of tracer appearance is easily measurable.

Journal Article↗
PubMed1994

[Formation of popular medical books in Sichuan Province in late Qing Dynasty and its influence].

Among the traditional Chinese medical works in the Qing Dynasty is Sichuan province, popular ones occupied the major part. This author investigates its causes from the angles of economics, population and characteristics of physicians in this province. He also discusses the abridgement or re-compilation on the basis of those authors by their own clinical experience, who either imitated the stkyle, recompiled, or even attached their own work to Chen Xiuyuan's medical books. The influence, significance and role of these medical books are also mentioned.

China↗
PubMed1993

Ca(2+)-induced folding and aggregation of skeletal muscle sarcoplasmic reticulum calsequestrin. The involvement of the trifluoperazine-binding site.

Calsequestrin is an intermediate affinity, high capacity Ca(2+)-binding protein found in the lumen of the sarcoplasmic reticulum of both skeletal and cardiac muscle cells. Previous sequence analysis suggested that calsequestrin may contain a hydrophobic binding site for the drug trifluoperazine, a site shared by the calmodulin family and shown to play a role in calmodulin/calmodulin receptor interaction. Previous studies showed that, upon Ca2+ binding, calsequestrin undergoes a conformational change, burying the trifluoperazine-binding site, folding into a more compact structure that is trypsin-resistant, and increasing the negative ellipticity of the circular dichroism spectrum. In this study, the structural and functional roles of the trifluoperazine-binding site in the Ca(2+)-induced conformational change of calsequestrin are further studied using the calmodulin antagonists trifluoperazine and melittin. If trifluoperazine or melittin is added to calsequestrin prior to Ca2+ addition, then Ca(2+)-induced folding is inhibited as determined by the changes in circular dichroism spectra and protein sensitivity to trypsin digestion. If, however, Ca2+ is added prior to trifluoperazine or melittin, calsequestrin remains resistant to trypsin digestion, just as if the calmodulin antagonists are not present, suggesting that the conformational change is not affected. Aggregates of calsequestrin that exhibit high Ca2+ binding capacity have previously been shown to occur at high Ca2+ and calsequestrin concentrations. By preventing a prerequisite folding step, trifluoperazine or melittin also prevents the Ca(2+)-induced aggregation of calsequestrin, thus decreasing the maximal Ca2+ binding by calsequestrin. These data suggest that the trifluoperazine-binding site is critically involved in the Ca(2+)-induced intramolecular folding step required for the intermolecular interactions leading to high capacity Ca(2+)-binding by calsequestrin.

Amino Acid Sequence↗