Biomedical subjects

X Wei

Publications and source records attributed to X Wei.

At least 91 records · Page 5Linked to original sources

PubMed1999

Pathogenesis of retinoic acid-induced ear malformations in primate model.

13-cis retinoic acid (RA) is a causative agent for human/monkey retinoic acid embryopathy (RAE), in which the most common type of malformation is microtia or anotia. In the present study, malformed ears of monkey fetuses exposed to RA during early embryogenesis were analyzed and revealed a subtype of defects., i.e., apparent duplication of the external/middle ear. A part of the posterior auricle appeared to be ectopically formed in the anterior auricular region or in the region posterior to the auricle. Additionally, there was duplication of the zygomatic arch, malleus, and incus. In order to characterize possible pathogenetic events underlying these malformations, embryos at selected stages were collected after dosing dams with RA at 5 mg/kg/day during gestational days 12-27. Cellular retinoic acid binding protein I whole-mount immunostaining showed that RA induced specific alterations in the migration of cranial neural crest cells (NCC). NCC en route to the second pharyngeal arch were bifurcated, and some of these NCC migrated abnormally into the first and/or third arches, which may underlie external ear duplication. Scanning electron microscopy and neurofilament immunostaining provided evidence that there was partial duplication of trigeminal nerve/ganglion following RA insult. The duplication of NCC neuronal derivatives in the first pharyngeal arch is consistent with duplication of NCC mesenchymal components (zygomatic arch, malleus, and incus). Therefore, RA-induced alterations in cranial NCC migration patterns are likely to be a pathogenetic event underlying ear malformations (including duplication) of RAE in monkeys.

Animals↗

PubMed1999

Tn5-induced and spontaneous switching of Sinorhizobium meliloti to faster-swarming behavior.

Tn5 mutants of Sinorhizobium meliloti RMB7201 which swarmed 1.5 to 2. 5 times faster than the parental strain in semisolid agar, moist sand, and viscous liquid were identified. These faster-swarming (FS) mutants outgrew the wild type 30- to 40-fold within 2 days in mixed swarm colonies. The FS mutants survived and grew as well as or better than the wild type under all of the circumstances tested, except in a soil matrix subjected to air drying. Exopolysaccharide (EPS) synthesis was reduced in each of the FS mutants when they were grown on defined succinate-nitrate medium, but the extent of reduction was different for each. It appears that FS behavior likely results from a modest, general derepression of motility involving an increased proportion of motile and flagellated cells and an increased average number of flagella per cell and increased average flagellar length. Spontaneous FS variants of RMB7201 were obtained at a frequency of about 1 per 10,000 to 20,000 cells by either enrichment from the periphery of swarm colonies or screening of colonies for reduced EPS synthesis on succinate-nitrate plates. The spontaneous FS variants and Tn5 FS mutants were symbiotically effective and competitive in alfalfa nodulation. Reversion of FS variants to wild-type behavior was sporadic, indicating that reversion is affected by unidentified environmental factors. Based on phenotypic and molecular differences between individual FS variants and mutants, it appears that there may be multiple genetic configurations that result in FS behavior in RMB7201. The facile isolation of spontaneous FS variants of Escherichia coli and Pseudomonas aeruginosa indicates that switching to FS behavior may be fairly common among bacterial species. The substantial growth advantage of FS mutants and variants wherever nutrient gradients exist suggests that switching to FS forms may be an important behavioral adaptation in natural environments.

Blotting, Southern↗

PubMed1999

Inhibition of human liver cytochrome P-450 1A2 by the class IB antiarrhythmics mexiletine, lidocaine, and tocainide.

Mexiletine, lidocaine, and tocainide are class IB antiarrhythmic drugs that are used for the treatment of ventricular arrhythmias and are known to inhibit drug metabolism. The objectives of this study were to characterize the inhibitory effects of mexiletine, lidocaine, and tocainide on cytochrome P-450 1A2 (CYP1A2) activity in human liver microsomes and to evaluate their relative inhibitory potencies by using a molecular model of this P-450 isozyme. The inhibitory effect of mexiletine, lidocaine, and tocainide on cytochrome CYP1A2 in human liver microsomes was examined with methoxyresorufin O-demethylase activity as an index of the catalytic activity of this P-450 isozyme. The kinetic inhibition types and Ki values were determined by Lineweaver-Burk plots and Dixon plots, respectively. Molecular modeling was used to assess the interaction of these agents with the CYP1A2 active site. Methoxyresorufin O-demethylase activity was inhibited 67 +/- 8%, 20 +/- 5%, and 7 +/- 4% by 2 mM mexiletine, lidocaine, and tocainide, respectively. Mexiletine and lidocaine exhibited competitive inhibition with Ki values of 0.28 +/- 0.12 mM and 1.54 +/- 0.74 mM, respectively, whereas the inhibition type of tocainide could not be determined because of its weak potency. A charge interaction between mexiletine and the Asp313 side chain in the CYP1A2 active site was found, and varying degrees of hydrogen bond formation between these three compounds and the CYP1A2 active site were observed. The in vitro inhibitory potencies in human liver microsomes (mexiletine > lidocaine > tocainide) are consistent with the structural interactions found in a molecular model of the active site of CYP1A2.

Anti-Arrhythmia Agents↗

PubMed1999

[Effect of Baoyuan Qiangshen capsule no. II on tubular interstitial injury in chronic renal failure patients].

OBJECTIVE: To investigate the protective effect and mechanism of Baoyuan Qiangshen Capsule No. II (BYQS) in treating chronic renal failure (CRF). METHODS: Sixty patients of CRF were divided into 2 groups randomly, the tested group treated with BYQS combined with Lotensin and the control group treated with essential amino acid combined with Lotensin. Changes of renal function and tubular marked proteins were observed. RESULTS: The markedly effective rate and total effective rate of the tested group were 63.3% and 93.3% respectively and those of the control group were 30.0% and 56.7% respectively, the effect of the tested group was markedly better than that of the control group (P < 0.01). In the tested group after treatment, BUN, SCr and CCr were improved significantly, urine retinol binding protein and beta 2-microglobulin and N-acetyl-beta-D-glucosamindase decreased significantly (P < 0.01), while Tamm-Horfau protein increased significantly (P < 0.05). CONCLUSION: BYQS could alleviate tubular interstitial injury significantly so as to improve the renal function and enhance the effective rate in treating CRF.

Adolescent↗

PubMed1999

[The characteristics and management of shotgun wounds of the extremities].

OBJECTIVE: To discuss the characteristics, classification and the problem of management for shotgun wounds of the extremities. METHODS: 82 shotgun wounds of the extremities, classified according to the modified rule form Sherman and Ordog's, were treated by "incision and drainage" initially, and were closed in primary, delay and later phase. The fractures were fixed by plaster, external fixator, simple internal fixator or plate fixation. Vascular injury was repaired by suture end-to-end or autograft. The injured nerves were treated in the primary or later period. RESULTS: The rate of infection was the highest in the primary closed wound, and the healing time was the longest in the later closed wound. The rate of infection was much higher in the fracture fixed by plate than the others. The results of nerve recovery in primary repair were not significantly different (except 2 case infection) from those in the later repair. CONCLUSIONS: A significant difference was noted between shotgun wound and other wound. Removal of wadding, prevention of fascial compartment syndrome, repeat planned exploration of wound, proper wound closing, external fixation of fracture, vascular debridement before anastomosis and delayed nerves repair contribute to a successful outcome in patients with extremity shotgun wounds.

Adolescent↗

PubMed1999

[Antagonistic effects of metallothionein on genetic damage caused by gamma-rays and mitomycin C].

OBJECTIVE: To study the antagonistic effects of metallothionein (Zn-MT)on genetic damage caused by gamma-rays and mitomycin C (MMC) in the liver of rabbits. METHODS: Micronucleus test and DNA strand breaks were determined in vitro and in vivo with single-cell gel electrophoresis assay to observe the effects of Zn-MT on the micronucleus formation and cleavage of DNA chain before and after gamma-rays radiation and MMC treatment. RESULTS: A dose of 664 microg/kg of Zn-MT could inhibit micronucleus formation in the bone marrow of the mice induced by 5 Gy gamma-rays radiation (P < 0.05). Concentrations of 10 and 50 microg/ml Zn-MT could antagonize the increase in bimicronucleus frequency in g12 cells induced by 1 and 3 Gy gamma-rays radiation and 0.3 microg/ml MMC treatment (P < 0.05 or P < 0.01). A dose of 50 microg/ml Zn-MT could reduce the DNA single strand breaks caused by 1 Gy gamma-rays radiation, but without statistical significance (P > 0.05). CONCLUSION: Zn-MT could antagonize the genetic damage to certain extent in the liver of the rabbits caused by gamma-rays radiation and MMC treatment.

Animals↗

PubMed1999

[Studies on DNA single strand breaks induced by sodium nitroprusside-nitric oxide donor].

OBJECTIVE: To study the effect of sodium nitroprusside (SNP), a nitric oxide (NO) donor, on DNA single strand breaks (SSBs). METHODS: A modified method was used to isolate and prepare the single cell suspension from the organs of mice. Alkaline single-cell gel electrophoresis (SCGE) was performed to examine DNA damage of the cells treated by SNP in vivo and in vitro. RESULTS: Treatment with 0.5 - 2.0 micromol/ml of SNP with S(9) for 1 h induced a concentration-dependent increase in DNA SSBs in g12 cells. Significant increase in DNA migration and comet frequency in the spleen, thymus and peritoneal macrophage were induced after intraperitoneal injection of SNP at a dose of 0.67 - 6.0 mg/kg. No obvious increase in DNA single strand breaks was observed in the liver, kidney and lung of the mice with same treatment. CONCLUSION: DNA SSBs could be induced by NO in some cells in vivo and in vitro. There was difference in sensitivity of various organs in the mice to NO. Cells of spleen and thymus and macrophage may be the important target cells of NO.

Animals↗

PubMed1998

Segregation of transcription and replication sites into higher order domains.

Microscopy shows that individual sites of DNA replication and transcription of mammalian nuclei segregate into sets of roughly 22 and 16 higher order domains, respectively. Each domain set displayed a distinct network-like appearance, including regions of individual domains and interdigitation of domains between the two networks. These data support a dynamic mosaic model for the higher order arrangement of genomic function inside the cell nuclei.

3T3 Cells↗

PubMed1998

Characterization of the human dihydropyrimidine dehydrogenase gene.

Dihydropyrimidine dehydrogenase (DPD) catabolizes endogenous pyrimidines and pyrimidine-based antimetabolite drugs. A deficiency in human DPD is associated with congenital thymine-uraciluria in pediatric patients and severe 5-fluorouracil toxicity in cancer patients. The dihydropyrimidine dehydrogenase gene (DPYD) was isolated, and its physical map and exon-intron organization were determined by analysis of P1, PAC, BAC, and YAC clones. The DPYD gene was found to contain 23 exons ranging in size from 69 bp (exon 15) to 961 bp (exon 23). A physical map derived from a YAC clone indicated that DPYD is at least 950 kb in length with 3 kb of coding sequence and an average intron size of about 43 kb. The previously reported 5' donor splice site mutation present in pediatric thymine-uraciluria and cancer patients can now be assigned to exon 14. All 23 exons were sequenced from a series of human DNA samples, and three point mutations were identified in three racial groups as G1601A (exon 13, Ser534Asn), A1627G (exon 13, Ile543Val), and G2194A (exon 18, Val732Ile). These studies, which have established that the DPYD gene is unusually large, lay a framework for uncovering new mutations that are responsible for thymine-uraciluria and toxicity to fluoropyrimidine drugs.

Alleles↗

PubMed1998

Starvation-Induced Changes in Motility, Chemotaxis, and Flagellation of Rhizobium meliloti

The changes in motility, chemotactic responsiveness, and flagellation of Rhizobium meliloti RMB7201, L5-30, and JJ1c10 were analyzed after transfer of the bacteria to buffer with no available C, N, or phosphate. Cells of these three strains remained viable for weeks after transfer to starvation buffer (SB) but lost all motility within just 8 to 72 h after transfer to SB. The rates of motility loss differed by severalfold among the strains. Each strain showed a transient, two- to sixfold increase in chemotactic responsiveness toward glutamine within a few hours after transfer to SB, even though motility dropped substantially during the same period. Strains L5-30 and JJ1c10 also showed increased responsiveness to the nonmetabolizable chemoattractant cycloleucine. Cycloleucine partially restored the motility of starving cells when added after transfer and prevented the loss of motility when included in the SB used for initial suspension of the cells. Thus, interactions between chemoattractants and their receptors appear to affect the regulation of motility in response to starvation independently of nutrient or energy source availability. Electron microscopic observations revealed that R. meliloti cells lost flagella and flagellar integrity during starvation, but not as fast, nor to such a great extent, as the cells lost motility. Even after prolonged starvation, when none of the cells were actively motile, about one-third to one-half of the initially flagellated cells retained some flagella. Inactivation of flagellar motors therefore appears to be a rapid and important response of R. meliloti to starvation conditions. Flagellar-motor inactivation was at least partially reversible by addition of either cycloleucine or glucose. During starvation, some cells appeared to retain normal flagellation, normal motor activity, or both for relatively long periods while other cells rapidly lost flagella, motor activity, or both, indicating that starvation-induced regulation of motility may proceed differently in various cell subpopulations.

Journal Article↗

PubMed1998

Evidence that HetR protein is an unusual serine-type protease.

The hetR gene plays a very important role in cell differentiation of heterocystous cyanobacteria. To understand the mechanism of the hetR gene product in regulation of heterocyst differentiation, the recombinant HetR protein (rHetR) was overproduced in Escherichia coli. Purified rHetR was unstable and degraded easily in solution. Phenylmethanesulfonyl fluoride, a serine-type protease inhibitor, prevented the degradation and was shown to modify covalently rHetR. Dansyl fluoride (DnsF), another serine-type protease inhibitor, also covalently modifies rHetR as shown by electrophoresis and electroblotting of the labeled rHetR and by MS. The labeling of rHetR with phenylmethanesulfonyl fluoride and DnsF was at the same site of rHetR and required Ca2+. S179N-rHetR, a mutant protein from strain 216 of Anabaena PCC 7120, which cannot differentiate heterocysts because of the mutation, was also overproduced and characterized. Although S170N-rHetR still can be labeled with DnsF, no proteolysis was observed, suggesting that Ser179 is involved in proteolytic activity. DnsF-labeled rHetR was digested with trypsin, and the labeled peptide was isolated and sequenced. The labeled peptide matches a sequence from HetR. These results show that HetR is a protease.

Amino Acid Sequence↗

PubMed1998

Facilitation by the beta2a subunit of pore openings in cardiac Ca2+ channels.

1. Single channel recordings were performed on the cardiac calcium channel (alpha1C) in order to study the effect of coexpression of the accessory beta2a subunit. On-cell patch clamp recordings were performed after expression of these channels in Xenopus oocytes. 2. The alpha1C subunit, when expressed alone, had similar single channel properties to native cardiac channels. Slow transitions between low and high open probability (Po) gating modes were found as well as fast gating transitions between the open and closed states. 3. Coexpression of the beta2a subunit caused changes in the fast gating during high Po mode. In this mode, open time distributions reveal at least three open states and the beta2a subunit favours the occupancy of the longest, 10-15 ms open state. No effect of the beta2a subunit was found when the channel was gating in the low Po mode. 4. Slow gating transitions were also affected by the beta2a subunit. The high Po mode was maintained for the duration of the depolarizing pulse in the presence of the beta2a subunit; while the alpha1C channel when expressed alone, frequently switched into and out of the high Po mode during the course of a sweep. 5. The beta2a subunit also affected mode switching that occurred between sweeps. Runs analysis revealed that the alpha1C subunit has a tendency toward non-random mode switching. The beta2a subunit increased this tendency. A chi2 analysis of contingency tables indicated that the beta2a subunit caused the alpha1C channel to gain 'intrinsic memory', meaning that the mode of a given sweep can be non-independent of the mode of the previous sweep. 6. We conclude that the beta2a subunit causes changes to the alpha1C channel in both its fast and slow gating behaviour. The beta2a subunit alters fast gating by facilitating movement of the channel into an existing open state. Additionally, the beta2a subunit decreases the slow switching between low and high Po modes.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1998

1H and 13C NMR studies of a truncated heme domain from Chlorella vulgaris nitrate reductase: signal assignment of the heme moiety.

A water soluble truncated heme domain (a tetramer of MW = 45 kDa) of the tetrameric nitrate reductase complex from the green alga Chlorella vulgaris has been overexpressed and purified. This truncated heme domain with four identical subunits has a high redox potential (midpoint potential E1/2 = +16 mV) as compared with other heme-containing flavoproteins. We have undertaken a determination of the detailed configuration of the heme moiety in order to understand the unique electrochemical property of the heme moiety of this enzyme. We report here the study of the heme prosthetic group of the truncated heme domain by the use of 2D 1H and 13C NMR techniques. A complete signal assignment of the heme has been achieved. Our observations suggest that the heme configuration is similar to that of the crystal structure of the membrane-bound bovine liver cytochrome b5.

Animals↗

PubMed1998

Inhibition of human cytochrome P450 1A2 by flavones: a molecular modeling study.

Cytochrome P450 1A2 metabolizes a number of important drugs, procarcinogens, and endogenous compounds. Several flavones, a class of phytochemicals consumed in the human diet, have been shown to differentially inhibit human P450 1A2-mediated methoxyresorufin demethylase. A molecular model of this P450 was constructed in order to elucidate the molecular basis of the P450-flavone interaction. Flavone and its 3,5,7-trihydroxy and 3,5,7-trimethoxy derivatives were docked into the active site to assess their mode of binding. The site is hydrophobic and includes several residues that hydrogen bond with substituents on the flavone nucleus. The binding interactions of these flavones in the modeled active side are consistent with their relative inhibitory potentials, namely 3,5,7-trihydroxylflavone > flavone > 3,5,7-trimethoxylflavone, toward P450 1A2-mediated methoxyresorufin demethylation.

Amino Acid Sequence↗

PubMed1998

Inhibition of methoxyresorufin demethylase activity by flavonoids in human liver microsomes.

Flavonoids are a class of dietary phytochemicals with anticarcinogenic properties. A series of ten structurally related flavonoids were evaluated for their effect on methoxyresorufin O-demethylase (MROD) activity in human liver microsomes. All compounds inhibited this cytochrome P450 1A2 (CYP1A2) mediated activity. 3,5,7-Trihydoxyflavone (galangin) was the most potent inhibitor, followed by 3-hydroxyflavone and flavone. The relative inhibitory potency of flavonoids is related to their structures. The results suggest that flavonoids may modulate pharmacological and toxicological effects mediated by CYP1A2.

Adult↗

PubMed1998

Dihydropyrimidine dehydrogenase pharmacogenetics in patients with colorectal cancer.

Individuals with a deficiency in the enzyme dihydropyrimidine dehydrogenase (DPD) may experience severe life-threatening toxicity when treated with 5-fluorouracil (5-FU). As routine measurement of enzyme activity is not practical in many clinical centres, we have investigated the use of DNA mutation analysis to identify cancer patients with low enzyme levels. We have identified two new mutations at codons 534 and 543 in the DPD cDNA of a patient with low enzyme activity and screened the DNA from 75 colorectal cancer patients for these mutations and the previously reported splice site mutation (Vreken et al, 1996; Wei et al, 1996). In all cases, DPD enzyme activity was also measured. The splice site mutation was detected in a patient (1 out of 72) with low enzyme activity whereas mutations at codons 534 (2 out of 75) and 543 (11 out of 23) were not associated with low enzyme activity. These studies highlight the need to combine DPD genotype and phenotype analysis to identify mutations that result in reduced enzyme activity.

Adult↗

PubMed1998

Dihydropyrimidine dehydrogenase pharmacogenetics in Caucasian subjects.

AIMS: Dihydropyrimidine dehydrogenase (DPD) catalyses the reduction of pyrimidines, including the anticancer agent 5-fluorouracil (5FU). Impaired 5FU degradation, through low DPD activity, has led to severe, life-threatening or fatal toxicity after administration of 5FU. Complete DPD deficiency is associated with the inherited metabolic disease thymine uraciluria. Several mutations in the gene encoding DPD have recently been identified, but the phenotype-genotype concordance of these alterations in the general population has not been reported. METHODS: Mononuclear cells were isolated from whole blood and DPD activity was determined after ex vivo incubation with 14C-5FU followed by h.p.1.c. analysis of 5FU metabolites. Analysis of mutations in the DPD gene at an exon splice site, codons 534, 543, and 732, and a deletion at base 1897 (deltaC1897) were performed in 30 subjects with the lowest and 30 subjects with the highest enzyme activity using PCR-RFLP. RESULTS: DPD activity was measured in 226 Caucasian subjects and was highly variable (range 19.1-401.4 pmol min(-1)mg(-1) protein). Mutations were frequently observed at codons 543 (allele frequency 28%), 732 (allele frequency 5.8%), and 534 (allele frequency 0.8%), but were not associated with low DPD activity. There were no splice site or deltaC1897 mutations found in this population. CONCLUSIONS: The five mutations analysed in this study are insufficient for identification of patients at risk for 5FU toxicity or thymine uraciluria. Both the splice site mutation and deltaC1897 are relatively rare in the general Caucasian population. Therefore, identification of further molecular alterations is required to facilitate the use of DPD analysis in genetic diagnosis and cancer therapeutics.

Alleles↗

PubMed1998

Age- and injury-dependent concentrations of transforming growth factor-beta 1 and proteoglycan fragments in rabbit knee joint fluid.

OBJECTIVE: The purpose of the study was to characterize maturation-related changes of TGF-beta 1 and proteoglycan fragment (PG) concentrations in joint fluid of healthy rabbit knees, and to investigate changes associated with osteochondral injury and spontaneous repair. METHODS: In 26 young (age range: 12-15 weeks), 26 adolescent (18-25 weeks), and 26 adult (33-44 weeks) New Zealand white rabbits, TGF-beta 1 and PG concentrations were analyzed in joint fluid samples which were taken before, and at several time intervals (max. 1 year) after creation of a osteochondral defect in the knee medial femoral condyle. At death, the characteristics of the regenerated tissue in the defect and any signs of degeneration of adjacent cartilage were recorded and graded. RESULTS: In preoperative samples, TGF-beta 1 and PG concentrations decreased with maturation (P < 0.01), and were moderately correlated (r = 0.51, P < 0.001). Shortly after trauma, the concentrations of both substances were found increased, which was followed by a decrease up to 3 months, and then again an increase up to 1 year. However, meanwhile PG concentrations had similar magnitude irrespective of age, TGF-beta 1 concentrations never reached similarly high levels in adulthood as in infancy or adolescence. The cartilage adjacent to the defect had more signs for degeneration in younger rabbits, and also osteophytes were more common in young than adult animals. CONCLUSION: The similar pattern for TGF-beta 1 and PG concentrations during postnatal maturation may reflect the stimulatory effect of TGF-beta 1 on proteoglycan synthesis. The higher TGF-beta 1 concentrations in younger animals may be a reason for their better healing capacity, but also for their higher susceptibility to osteoarthritic change compared to adult animals.

Aging↗