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Biomedical subjects

Y Xu

Publications and source records attributed to Y Xu.

At least 847 records · Page 47Linked to original sources
PubMed1997

Genomic organization of the integrin beta 4 gene (ITGB4): a homozygous splice-site mutation in a patient with junctional epidermolysis bullosa associated with pyloric atresia.

Integrin beta 4 is expressed primarily within the epithelial basement membranes, and it contributes to the stable association of epidermis with the underlying basement membrane. Previous observations have suggested that the expression of this integrin, which is alternatively spliced in various cell types, is deficient in a variant of junctional epidermolysis bullosa associated with pyloric atresia (JEB-PA). To facilitate identification of mutations in the human beta 4 integrin gene, ITGB4, we have now determined its intron-exon organization. The entire gene was shown to consist of 41 exons spanning 36 kb of the genomic DNA on chromosome 17q11-qter. The mutation detection, using PCR-amplification of each exon followed by heteroduplex analysis, revealed a homozygous splicing mutation in a patient with JEB-PA. RT-PCR revealed the presence of two splice variants generated through utilization of cryptic splice sites, and both mRNA transcripts resulted in a frame-shift and premature termination codon of translation. The presence of the mutation resulted in dramatic reduction of the corresponding mRNA transcript level. Because beta 4 integrin is expressed not only in the skin but also in the epithelial lining of the stomach, the absent expression of this integrin in the proband may explain the blistering tendency and development of pyloric atresia.

Amino Acid Sequence↗
PubMed1997

Cryptic and regulatory epitopes in CD13/aminopeptidase N.

CD13/Aminopeptidase N (APN) is a ubiquitous cell surface enzyme thought to be involved in downregulation of regulatory peptide signals, in binding viruses, and possibly in tumor invasion. Functional aspects of CD13/APN were probed using two monoclonal antibodies (F23 and MY7) to two distinct epitopes on the protein and using substrates and inhibitors of the enzyme. F23 was capable of completely blocking, and MY7 was capable of partially blocking, the binding of substrate to APN and the enzymatic activity of APN. The number of epitopes on APN was quantitated by flow cytometry and by radiobinding with Scatchard analysis. There were approximately 20,000 F23 epitopes per HL60 cell, whereas there were about 10,000 MY7 epitopes per cell. After binding of F23 or natural peptide substrates, the number of MY7 epitopes increased to become equimolar with F23 epitopes, showing a conformational change in CD13/APN structure that reveals MY7 epitopes. Mild proteinase treatment also revealed the hidden epitopes and equalized epitope numbers. The presence of cryptic epitopes may explain the different effects of these antibodies on substrate binding and enzymatic activity. Competition analysis identified the substrate binding site as involving the zinc binding domain of the enzyme. The substrates blocked F23 epitopes (zinc binding domain of the enzyme) and MY7 epitopes, whereas inhibitors blocked only the F23 epitopes, and downregulated MY7 epitopes. A dimeric structure for CD13/APN, in its native, membrane-bound state was directly demonstrated by cross-linking with Bis (sulfosuccinimidyl) suberate and gel electrophoresis. Despite the conformational changes that increased or decreased MY7 epitopes in CD13/APN after substrate or inhibitor binding, neither substrates nor inhibitors caused alterations in the ratio of homodimers to monomers. This suggests that APN undergoes intramolecular conformational changes rather than gross separation of protomers during its regulation. A model for the conformational regulation of APN is proposed.

Antibodies, Monoclonal↗
PubMed1997

Microsatellite marker development, mapping and applications in rice genetics and breeding.

Microsatellites are simple, tandemly repeated di- to tetra-nucleotide sequence motifs flanked by unique sequences. They are valuable as genetic markers because they are co-dominant, detect high levels of allelic diversity, and are easily and economically assayed by the polymerase chain reaction (PCR). Results from screening a rice genomic library suggest that there are an estimated 5700-10,000 microsatellites in rice, with the relative frequency of different repeats decreasing with increasing size of the motif. A map consisting of 120 microsatellite markers demonstrates that they are well distributed throughout the 12 chromosomes of rice. Five multiple copy primer sequences have been identified that could be mapped to independent chromosomal locations. The current level of genome coverage provided by these simple sequence length polymorphisms (SSLPs) in rice is sufficient to be useful for genotype identification, gene and quantitative trail locus (QTL) analysis, screening of large insert libraries, and marker-assisted selection in breeding. Studies of allelic diversity have documented up to 25 alleles at a single locus in cultivated rice germplasm and provide evidence that amplification in wild relatives of Oryza sativa is generally reliable. The availability of increasing numbers of mapped SSLP markers can be expected to complement existing RFLP and AFLP maps, increasing the power and resolution of genome analysis in rice.

Breeding↗
PubMed1997

Inferring gene structures in genomic sequences using pattern recognition and expressed sequence tags.

Computational methods for gene identification in genomic sequences typically have two phases: coding region prediction and gene parsing. While there are many effective methods for predicting coding regions (exons), parsing the predicted exons into proper gene structures, to a large extent, remains an unsolved problem. This paper presents an algorithm for inferring gene structures from predicted exon candidates, based on Expressed Sequence Tags (ESTs) and biological intuition/rules. The algorithm first finds all the related ESTs in the EST database (dbEST) for each predicted exon, and infers the boundaries of one or a series of genes based on the available EST information and biological rules. Then it constructs gene models within each pair of gene boundaries, that are most consistent with the EST information. By exploiting EST information and biological rules, the algorithm can (1) model complicated multiple gene structures, including embedded genes, (2) identify falsely-predicted exons and locate missed exons, and (3) make more accurate exon boundary predictions. The algorithm has been implemented and tested on long genomic sequences with a number of genes. Test results show that very accurate (predicted) gene models can be expected when related ESTs exist for the predicted exons.

Algorithms↗
PubMed1997

Hybridization dominance of kinetics in recombinant ATH-1376 obtained via protoplast fusion between Aspergillus niger and Trichoderma reesei.

Comparisons of the kinetics of mycelium growth, cellulase biosynthesis, and the degradation of filter paper to accumulate reducing-sugar by the filtrates of cultures were carried out among the recombinant strain ATH-1376 and its two parents, Aspergillus niger AMS11 and Trichoderma reesei QM9414. The results showed that both the specific mycelium growth rate and the cellulase biosynthesis rate of the recombinant were dramatically dominant over those of the two parents. In addition, the negative correlation between the specific mycelium growth rate and the cellulase biosynthesis rate of the recombinant ATH-1376 was much lower than those of its parents. In terms of the amount of reducing-sugar accumulated from the hydrolysis of filter paper by culture filtrates, there were great differences among the three different treatments, i.e., fermentation filtrate of single parental strain, mixture of the fermentation filtrates from two parental strains with different ratios (v:v), and filtrate from the mixed culture of the two parental strains. Out of these, the second approach, particularly with the ratio of 1:1, was best for the accumulation of reducing sugar. Within various tested periods of enzymic hydrolysis, the amounts of reducing-sugar produced by the recombinant were 1.19 to 2.26 times as much as the maximum amounts produced in parallel by the mixture of filtrates (1:1) from separate fermentations of the two parental strains. These results suggested that constructing the engineered strains with hybridization dominance of these two typical genera of far-heredity could be effective to overcome the great deficiencies of routine mixculture, single-strain fermentation, or double fed-batch fermentations.

Aspergillus niger↗
PubMed1997

Long-term prognosis of percutaneous transluminal coronary angioplasty in 229 cases.

OBJECTIVE: To evaluate long-term efficacy of percutaneous transluminal coronary angioplasty (PTCA) and the factors affecting its efficacy in Chinese patients. METHODS: Two hundred and twenty-nine patients who underwent successful PTCA in this institution were followed-up by direct interview or letters. The rate of follow-up was 95%. The period of follow-up were 0.5-8.4 (with mean of 2.3 +/- 1.8 years). RESULTS: Angina Pectoris recurred in 76 (33.2%) of the patients. Cox regression analysis revealed that the relative risk of recurrence of angina pectoris was increased among the patients with triple vessel disease and with history of hypertension. During the period of follow-up, 2 (0.9%) patients died, 6 (2.6%) had non-fatal acute myocardial infarction, 4 (1.7%) had coronary artery bypass graft surgery and 29 (12.7%) had repeat PTCA. The cardiac event-free survival rate calculated by Kaplan-Meier method was 84.8% at the first year and 70.5% at the eighth year. Cox regression analysis revealed that there were positive correlation between the relative risks of cardiac events and stenosis of lesions before PTCA and residual stenosis of LAD after PTCA. CONCLUSION: The long-term efficacy of PTCA in Chinese patients was good. The results of this study suggest that to decrease the residual stenosis of LAD during procedures could probably decrease the relative risk of cardiac events during follow-up.

Adult↗
PubMed1997

[Comparison of stature growth curves in children and adolescents].

Growth charts of height in Chinese children and adolescents (excluding Taiwan and Hong Kong) in the 1980s and 1990s were fitted to JPA2 model to study their growth and developmental characteristics and their future trend and to compare them with the criteria for growth in the developed countries and areas. Results showed that fitted growth curves were comparable. The difference in body height of children in cities during puberty period was the most significant during the 1980s. Body height of boys and girls aged over seven to maturity period in Beijing during the 1990s was higher than that in Hong Kong and Japan, and reached basically the criteria set by the National Center for Health Statistics (NCHS), United States, but body height of adults was still three to four centimeters lower than that in criteria set by NCHS. Body height in adolescents in Beijing was kept in stable as compared with that in the 1980s.

Adolescent↗
PubMed1997

[New species of Natronobacterium].

An extreme haloalkalophilic bacterium HAM-2 with pleomorphic rods was isolated from Hamatai soda lake in Inner Mongolia Autonomous Region. Growth occurs in 12%-30% NaCl, no growth below 12%, optimum 17.5%. pH range for good growth 7.8-10.4, optimum 9.0-9.5. The strain HAM-2 is non-motile, Gram-negative, pleomorphic, cell of which were 1.0-2.0 by 2.0-5.0 microns in size in liquid medium. The major polar lipids of the organisms are phosphatidylglycerol phosphate and phosphatidyglycerol and contains an unidentified phospholipid as a minor component PL4. G + C contant of DNA is 59.5 mol%. Based on the characteristics, the strain HAM-2 could be included in the genus Natronobacterium, but since HAM-2 differs from reported four species of this genus in cell shape, polar lipids and biochemical features. The name Natronobacterium innemongoliae sp. nov. is proposed. The type strain is designated HAM-2.

Natronobacterium↗
PubMed1997

[Repair of pulp defect of thumb by free palmaris brevis musculocutaneous flap].

It is difficult to repair the pulp defect of finger with good function. Here reported two cases of pulp defect of the thumb which were repaired with free palmris brevis musculo-cutaneous flap. The flap was designed as the following: taken the line crossing the pisiform and metacarpo-phalangeal joint of the little finger as the longitudinal axis of the flap. The proximal end of the flap was at the level of pisiform and the distal end was the distal transverse palmar crease. The radial border was the radial side of the palmris brevis and ulnar border was the ulnar edge of the palm. The flap should not be larger than 6.5 cm x 2.5 cm. The flap was dissected with proper ulnar vessels and the accompanying arterial branch and superficial branch of the ulnar nerve. The flap was transferred to repair the pulp defect of the thumb. The vessels were anastomosed with the radial artery and cephalic vein at the snuffbox. The nerves were sutured to the digital nerve. Three months after operation, the defect was healed and the sensation of pulp was recovered. The donor area was painless and without sensation disturbance. The motion of the little finger was normal. The function of the thumb was restored. It was concluded that the palmaris brevis musculocutaneous flap is one of the best donor region to repair pulp defect of thumb.

Adult↗
PubMed1997

[Repair and functional reconstruction of war wounds of limbs].

From 1979 to 1994, reparative and reconstructive surgery were used to repair the war injuries of skins, bones, blood vessels and nerves of the limbs in 800 cases. A systematic clinical study was carried out. Many new operative methods were used and the results of treatment were good. Innovations and modifications were made in technique. In 120 cases of war injuries having soft tissues defects including skin and muscles, various tissue transplantations were used with the hope to accomplish one-staged repair of the defect and reconstruction of motor function of muscle. To those infections of bone and joint in war injuries, following early eradication of infected focus, transplantation of musculo-cutaneous flap or omental graft was immediately carried out with the aim to obtain primary healing of the wound. In the treatment of bone defects from war wounds with loss of skin and muscles, the vascularized skeleto-cutaneous graft was used. In the treatment of 150 cases of injury of peripheral nerve from forearms, the result of good to fair rated 68.8 percent for upper extremity and that for lower extremity, it was 62.2 percent. Following the early repair of 500 cases of injury of peripheral blood vessels, the patency rate of the blood vessel was 90 percent. The result following by pass vascular graft in the treatment of forearms injury of blood vessels even with very poor local condition was still very successful.

Adolescent↗
PubMed1996

Serotonin transporter antibodies: production, characterization, and localization in the brain.

Serotonin (5-HT) transporter, the mechanism for 5-HT high affinity uptake, is the essential component for the termination of 5-HT transmission. In order to identify transporter sites on 5-HT neurons or on other 5-HT uptaking cells, three rabbit antisera against cocaine sensitive-serotonin transporter (5-HTT) were produced. Antisera 5-HTT55 (against amino acid sequence 55-68 in cytoplasmic N-terminal) and 5-HTT315 (against amino acid sequence 315-325, a 3rd external loop peptide) were produced against synthetic multiple-antigenic peptides (MAP). Antiserum 5-HTTN was produced against a fusion protein of the first 71 amino acids of N-terminal peptide expressed in recombinant DNA transformed bacteria. SDS-PAGE/Western blots indicate that 5-HTT55 and 5-HTT315 recognized bands of 74 and 64 kDa in rat brains with densities in the order of cortex > or = hippocampus > cerebellum, but not in liver, or muscle. The 5-HTTN recognized the fusion protein expressed in the bacteria, and the 64 kDa band with a similar density profile in the rat brain regions, and negative in liver and muscle. The immunocytochemistry of all three antisera revealed 5-HTT-immunostaining (5-HTT-im) in a pattern similar to 5-HT fiber distribution. 5-HTT55 and 5-HTT315 stainings were punctate in appearance, while 5-HTTN outlined the fibers in the 5-HT fiber areas, and neurons in raphe but not in substantia nigra or locus ceruleus. The preimmune serum and immune serum preabsorbed with 5-HTTN showed negative or diminished staining. Specific neurotoxin, 5,7-dihydroxytryptamine lesion removed all of the 5-HTTN fibers from the injection site, indicating, that 5-HTTN-im fibers are 5-HT fibers in nature. Our study indicates that the three antibodies we produced recognize various domains of the 5-HTT. Our 5-HTT antibodies could be used as new markers of 5-HT fibers, and are particularly useful for the study of the plasticity of 5-HT fibers free of the complications involved with 5-HT content.

Amino Acid Sequence↗
PubMed1996

The faciogenital dysplasia gene product FGD1 functions as a Cdc42Hs-specific guanine-nucleotide exchange factor.

The Rho family of small GTP-binding proteins plays important roles in the regulation of actin cytoskeleton organization and cell growth. Activation of these GTPases involves the replacement of bound GDP with GTP, a process catalyzed by the Dbl-like guanine-nucleotide exchange factors, all of which seem to share a putative catalytic motif termed the Dbl homology (DH) domain, followed by a pleckstrin homology (PH) domain. Here we have examined the role of a Dbl-like molecule, the faciogenital dysplasia gene product (FGD1), which when mutated in its Dbl homology domain, cosegregates with the developmental disease Aarskog-Scott syndrome. We report that a polypeptide of FGD1 encompassing the DH and PH domains can bind specifically to the Rho family GTPase Cdc42Hs and stimulates the GDP-GTP exchange of the isoprenylated form of Cdc42Hs. Microinjection of this FGD1 polypeptide into Swiss 3T3 fibroblast cells induces the formation of peripheral actin microspikes, similar to that previously observed when cells were injected with a constitutively active form of Cdc42Hs. This effect of FGD1 on actin organization is readily inhibited by coinjection of a dominant-negative mutant of Cdc42Hs. Examination of NIH 3T3 cells expressing the FGD1 fragment revealed that similar to cells expressing Dbl, two independent elements downstream of Cdc42Hs, the Jun NH2-terminal kinase and the p70 S6 kinase, became activated. Hence, our results indicate that FGD1, through its DH and PH domains, acts as a Cdc42Hs-specific guanine-nucleotide exchange factor and suggest that the Cdc42Hs GTPase may have a role in mammalian development.

3T3 Cells↗
PubMed1996

The Conductivity Confined Temperature Dependence of Water-Free Electrorheological Fluids

The shear stresses and the current densities of four sorts of water-free ER fluids, two oxidized polyacrylonitriles-based and two aluminosilicates-based, were investigated at elevated and reduced temperatures in order to elucidate the operating mechanism. The complicated relationships among the mechanical properties, the current density, and temperature were especially addressed. The Wagner model was introduced to interpret our observations, instead of the commonly used conductivity model. It is found that the conductivity of the dispersed particles determines the temperature dependence of the shear stress and the current density of an ER fluid. Our findings have direct implication for the design of industrially required ER fluids and would be significant for ER applications.

Journal Article↗
PubMed1996

Antineutrophil cytoplasmic autoantibodies interact with primary granule constituents on the surface of apoptotic neutrophils in the absence of neutrophil priming.

The pathogenic role of antineutrophil cytoplasmic autoantibodies (ANCA) remains controversial because of the difficulty in explaining how extracellular ANCA can interact with intracellular primary granule constituents. It has been postulated that cytokine priming of neutrophils (PMN), as may occur during a prodromal infection, is an important trigger for mobilization of granules to the cell surface, where they may interact with ANCA. We show by electron microscopy that apoptosis of unprimed PMN is also associated with the translocation of cytoplasmic granules to the cell surface and alignment just beneath an intact cell membrane. Immunofluorescent microscopy and FACS analysis demonstrate reactivity of ANCA-positive sera and antimyeloperoxidase antibodies with apoptotic PMN, but not with viable PMN. Moreover, we show that apoptotic PMN may be divided into two subsets, based on the presence or absence of granular translocation, and that surface immunogold labeling of myeloperoxidase occurs only in the subset of PMN showing translocation. These results provide a novel mechanism that is independent of priming, by which ANCA may gain access to PMN granule components during ANCA-associated vasculitis.

Antibodies, Antineutrophil Cytoplasmic↗
PubMed1996

Dual roles of ATM in the cellular response to radiation and in cell growth control.

The gene mutated in ataxia-telangiectasia (AT) patients, denoted ATM, encodes a putative protein or lipid kinase. To elucidate the functions of ATM, we disrupted the mouse ATM gene through homologous recombination in mice. Consistent with cellular defects of AT patients, the ATM-/- cells are hypersensitive to gamma-irradiation and defective in cell-cycle arrest following radiation, correlating with a defective up-regulation of p53. In addition, ATM-/- mouse thymocytes are more resistant to apoptosis induced by gamma-irradiation than normal thymocytes. ATM-/- fibroblasts are inefficient in G1 to S-phase progression following serum stimulation and senesce after only a few passages in culture. They have an increased constitutive level of p21CP1/WAF1. The ATM protein is therefore critical both for cellular responses to ionizing radiation and for normal cell-cycle progression. ATM+/- fibroblasts and thymocytes showed intermediately defective responses to irradiation but no growth defect, suggesting that the increased cancer risk of AT heterozygotes could be attributable to poor checkpoint function.

Animals↗
PubMed1996

Targeted disruption of ATM leads to growth retardation, chromosomal fragmentation during meiosis, immune defects, and thymic lymphoma.

ATM, the gene mutated in the inherited human disease ataxia-telangiectasia, is a member of a family of kinases involved in DNA metabolism and cell-cycle checkpoint control. To help clarify the physiological roles of the ATM protein, we disrupted the ATM gene in mice through homologous recombination. Initial evaluation of the ATM knockout animals indicates that inactivation of the mouse ATM gene recreates much of the phenotype of ataxia-telangiectasia. The homozygous mutant (ATM-/-) mice are viable, growth-retarded, and infertile. The infertility of ATM-/- mice results from meiotic failure. Meiosis is arrested at the zygotene/pachytene stage of prophase I as a result of abnormal chromosomal synapsis and subsequent chromosome fragmentation. Immune defects also are evident in ATM-/- mice, including reduced numbers of B220+CD43- pre-B cells, thymocytes, and peripheral T cells, as well as functional impairment of T-cell-dependent immune responses. The cerebella of ATM-/- mice appear normal by histologic examination at 3 to 4 months and the mice have no gross behavioral abnormalities. The majority of mutant mice rapidly develop thymic lymphomas and die before 4 months of age. These findings indicate that the ATM gene product plays an essential role in a diverse group of cellular processes, including meiosis, the normal growth of somatic tissues, immune development, and tumor suppression.

Animals↗