Biomedical subjects

K Ying

Publications and source records attributed to K Ying.

At least 19 recordsLinked to original sources

PubMed2004

Dermal fibroblast-associated gene induction by asiaticoside shown in vitro by DNA microarray analysis.

BACKGROUND: Asiaticoside, isolated from Centella asiatica, promotes fibroblast proliferation and extracellular matrix (ECM) synthesis in wound healing. The precise mechanism, however, in molecular and gene expression levels is still unclear. OBJECTIVE: Using cDNA microarray technology, the alteration of gene expression profiles was determined for human dermal fibroblasts in vitro in the presence of asiaticoside (30 microg mL(-1)). Fifty-four genes, with known functions for cell proliferation, cell cycle progression and synthesis of ECM, were significantly upregulated in our 'genome-nest' expression profile at various time points. Furthermore, the mRNA levels and protein production of certain genes responsible for ECM synthesis (e.g. encoding type I and type III collagen proteins) were evaluated by Northern blot and radioimmunoassay, respectively. RESULTS: We found that there is a close correlation between the gene profile, mRNA and protein production in the response of the cells to asiaticoside stimulation. CONCLUSIONS: This information could be used for exploring the response of the target genes to asiaticoside in fibroblasts.

Blotting, Northern↗

PubMed2003

Adaptive approximation of the boundary surface of a neuron in confocal microscopy volumetric images.

In biomedical visualisation, the isosurface is usually used to represent (approximate) the boundary surface of the structure within biomedical volumetric images. However, in many confocal microscopy volumetric images of neurons, the grey values of the object and/or background are usually uneven. Therefore a fixed isosurface is not suitable for use in approximating the boundary surface of the neuron. A method is proposed to construct the adaptively approximating surface of the boundary surface of the neuron. In this method, the boundary surface of the neuron could be locally and adaptively approximated with different surface patches in different local regions. Consequently, the approximation accuracy has been considerably improved.

Algorithms↗

PubMed2003

A human homolog of the yeast gene encoding tRNA 2'-phosphotransferase: cloning, characterization and complementation analysis.

The Saccharomyces cerevisiae TPT1 gene plays a role in removing the 2'-phosphate from ligated tRNA during the maturation of pre-tRNA. Here we reported the cloning and characterization of the human TRPT1 gene as a homolog of yeast TPT1. The TRPT1 gene is located at human chromosome 11q13 and encodes a polypeptide of 253 amino acids. BLAST searches with its amino acid sequence revealed the ubiquitous occurrence of TRPT1 homologs and their functional relationships with the presence of the DUF60/KptA domain. Northern analysis demonstrated that the gene is primarily expressed in heart and skeletal muscle, with lower or undetectable levels in other tissues studied. A plasmid-shuffling experiment showed that the human TRPT1 gene could complement the tpt1 mutation in S. cerevisiae

Amino Acid Sequence↗

PubMed2002

Isolation and characterization of a human novel RAB (RAB39B) gene.

Rab proteins are small-molecular-weight GTPases that control vesicular trafficking in eukaryotic cells. During the large-scale sequencing analysis of a human fetal brain cDNA library, we isolated a cDNA clone encoding a novel Rab protein, which showed 74.2% identity with previously isolated Rab39A at the amino acid level. RAB39B was expressed in a variety of human tissues and located in human chromosome Xq28. It consisted of two exons spanning 3764 bp of human genomic DNA.

Adult↗

PubMed2001

Identification of dopamine responsive mRNAs in glial cells by suppression subtractive hybridization.

Recent studies have established that glial cells are important targets of the neurotransmitter dopamine (DA), but the regulatory effects of DA on glial cells have not been extensively studied. In the present study, we have investigated the influence of DA on gene transcription in glial cells. Two-directional (forward and backward) suppression subtraction hybridization (SSH) was performed on astrocytes cultured from rat cerebral tissues in standard media or in culture media treated with DA. PCR-select differential screening was used to further verify the differentially expressed cDNA clones, positive clones were sequenced, and the mRNAs were re-examined on Northern blots. Fourteen sequences were identified of which eleven are homologous to known genes, three are homologous to expressed sequence tags (ESTs). Three novel full-length cDNAs were isolated using the EST fragments as probes to screen a cDNA library constructed from human brain. Analysis of these sequences suggested that complex intracellular signaling pathways, involving crosstalk with growth factor pathways, steroid hormone pathways, and an interferon-regulated 2-5 A pathway, are responsive to DA in astrocytes. The responsive proteins downstream from the signaling pathways were found to fall into at least three groups, including a series of metabolic enzymes, stress proteins, transfer proteins, etc. In addition, several of them have established their relationships with specific neurodegenerative diseases, showing that there is overlap in the pathogenic mechanisms of different diseases. Our results have provided a foundation for better understanding of the molecular basis of glial cell functions in dopaminergic transmission and an approach to find possible medication for the related disorders.

Animals↗

PubMed2001

Cloning and expression pattern of the human NDRG3 gene.

We report the cloning and expression pattern of a novel N-myc downstream-regulated gene 3 (NDRG3), located on human chromosome 20q11.21-11.23. The NDRG3 cDNA is 2588 base pair in length, encoding a 363 amino acid polypeptide highly related to mouse Ndr3 protein. Northern blot reveals that NDRG3 is highly expressed in testis, prostate and ovary. By in situ hybridization, the NDRG3 mRNA was localized to the outer layers of seminiferous epithelium, indicating that it may play a role in spermatogenesis.

Amino Acid Sequence↗

PubMed2001

Molecular cloning, mapping and characterization of the human neurocalcin delta gene (NCALD).

We identified a new human gene that encodes a cognate of the bovine neurocalcin delta from a human fetal brain cDNA library; hence we named it human neurocalcin delta (NCALD) gene. The deduced polypeptide product of the cDNA is 22 kDa in size, and its amino acid sequence is 100% and 99% identical to that of the bovine and chicken neurocalcin, respectively. Northern blots showed that the NCALD gene is more abundantly expressed in brain, testis, ovary and small intestine. Tissue in situ hybridization confirmed the existence of the NCALD mRNA in the adult human testis. Radiation hybrid panel mapping localized the gene to chromosome 8 between molecular markers D8S270 and D8S257.

Adult↗

PubMed2001

Cloning, expression, and characterization of a human inosine triphosphate pyrophosphatase encoded by the itpa gene.

ITP and dITP exist in all cells. dITP is potentially mutagenic, and the levels of these nucleotides are controlled by inosine triphosphate pyrophosphatase (EC ). Here we report the cloning, expression, and characterization of a 21.5-kDa human inosine triphosphate pyrophosphatase (hITPase), an enzyme whose activity has been reported in many animal tissues and studied in populations but whose protein sequence has not been determined before. At the optimal pH of 10.0, recombinant hITPase hydrolyzed ITP, dITP, and xanthosine 5'-triphosphate to their respective monophosphates whereas activity with other nucleoside triphosphates was low. K(m) values for ITP, dITP, and xanthosine 5'-triphosphate were 0.51, 0.31, and 0.57 mm, respectively, and k(cat) values were 580, 360, and 640 s(-1), respectively. A divalent cation was absolutely required for activity. The gene encoding the hITPase cDNA sequence was localized by radiation hybrid mapping to chromosome 20p in the interval D20S113-D20S97, the same interval in which the ITPA inosine triphosphatase gene was previously localized. A BLAST search revealed the existence of many similar sequences in organisms ranging from bacteria to mammals. The function of this ubiquitous protein family is proposed to be the elimination of minor potentially mutagenic or clastogenic purine nucleoside triphosphates from the cell.

Amino Acid Sequence↗

PubMed2001

Cloning and characterization of a novel human leptin receptor overlapping transcript-like 1 gene (LEPROTL1).

A new full-length cDNA encoding a novel protein was isolated from our human fetal brain cDNA library. The cDNA consists of 2701 bp and has a putative open reading frame encoding 131 amino acids which possesses a JAK binding site (Pro(46)-Ile-Pro(48) which is preceded by a cluster of hydrophobic residues) and is highly homologous to the leptin receptor gene-related protein (OB-RGRP). Northern blot analysis showed that this new gene is widely expressed in human tissues and radiation hybrid mapping placed the gene to human chromosome 8p21.1-8p21.2.

Amino Acid Sequence↗

PubMed2001

Cloning and characterization of a human lysyl oxidase-like 3 gene (hLOXL3).

Using the PCR primers generated from human expressed sequence tag (EST), the cDNA of lysyl oxidase-like gene 3 (LOXL3), a new member of human lysyl oxidases gene family, was cloned from the human fetal brain mRNA. The predicted amino acid sequence of the hLOXL3 gene was highly homologous to mLOR2. Bioinformatics analysis shows that hLOXL3 protein is also a member of the scavenger receptor cysteine-rich family, which contains a 25 amino acids signal peptide. The hLOXL3 gene was mapped to human 2p13 locus by BLAST search and at least 14 exons were found. Expression of the hLOXL3 gene was detected in several human tissues and especially high in spleen and testis.

Amino Acid Oxidoreductases↗

PubMed2001

PDZ-binding kinase participates in spermatogenesis.

A cDNA clone denoted SPK (spermatogenesis-related protein kinase) was isolated from human fetal brain cDNA library. The deduced protein was 99% identical to human PDZ-binding kinase (PBK), which is a mitotic kinase. Its isoform, which is about 300 bp longer at 3' end, was also isolated in this study. SPK gene was assigned to human chromosome 8p21.2 by radiation hybrid. Northern blot analysis indicated that SPK mRNA was predominantly expressed in testis. SPK mRNA was localized to the outer cell layer of seminiferous tubules by in situ hybridization, suggesting involvement of it in the process of spermatogenesis.

Amino Acid Sequence↗

PubMed2001

Cloning and characterization of KLHL5, a novel human gene encoding a kelch-related protein with a BTB domain.

Most kelch family proteins contain two conserved domains, the BTB domain and the kelch repeat domain. Here we describe the cloning and characterization of a novel human KLHL5 gene. The 3488 bp cDNA encodes a kelch family protein homologous to the Drosophila kelch protein. It also contains the two conserved domains. Northern blot analysis reveals a single transcript. It is abundantly expressed in ovary, adrenal gland, and thyroid and less abundantly expressed in trachea, prostate, testis, lymph node, and spinal cord. KLHL5 was mapped to 4p13-4p15.1.

Amino Acid Sequence↗

PubMed2001

Identification of a human LNX protein containing multiple PDZ domains.

Recently PDZ domains have been recognized as crucial organizers of protein complexes at the plasma membrane. Here we report the cloning of a novel 3.7-kb cDNA LNX from a human fetal brain cDNA library. The deduced amino acid sequence shares about 88% identity with the p70 form of Lnx, which is the mouse ligand of Numb protein X. Motif analysis reveals five protein-interaction modules conservative to those in the Lnx protein, including four PDZ domains and a NPXY motif for binding of the Numb PTB domain. Northern blot analysis shows that the length of the transcript in brain is distinct from those in other tissues. By radiation hybrid mapping, we localize the LNX gene to human chromosome 4q12 between marker D4S1577 and marker D4S1594. We predict that LNX may be important for the organization of signaling complexes in Numb-involved pathways or pathways regulated by other PTB-containing proteins.

Amino Acid Motifs↗

PubMed2001

Molecular cloning and characterization of a novel Dehydrogenase/reductase (SDR family) member 1 genea from human fetal brain.

Short-chain dehydrogenases/reductases (SDR) constitute a large protein family of NAD(P)(H)-dependent oxidoreductase. They are defined by distinct, common sequence motifs and show a wide range of substrate specialisms. By large-scale sequencing analysis of a human fetal brain cDNA library, we isolated a novel human SDR-type dehydrogenase/reductase gene named Dehydrogenase/reductase (SDR family) member 1 (DHRS1). The DHRS1 cDNA is 1411 base pair in length, encoding a 314-amino-acid polypeptide which has a SDR motif. Northern blot reveals two bands, of about 0.9 and 1.4 kb in size. These two forms are expressed in many tissues. The DHRS1 gene is localized on chromosome 14q21.3. It has 9 exons and spans 9.2 kb of the genomic DNA.

Amino Acid Sequence↗

PubMed2001

A novel human hydroxysteroid dehydrogenase like 1 gene (HSDL1) is highly expressed in reproductive tissues.

We report the cloning and characterization of a novel human hydroxysteroid dehydrogenase like gene (HSDL1) located on human chromosome 16q24.2. The HSDL1 cDNA is 3407 base pair in length, encoding a 309 amino acid polypeptide related to human 17beta-HSD3. Northern blot reveals that the HSDL1 is highly expressed in testis and ovary. In situ hybridization indicates that the expression of HSDL1 is predominantly increased in the prostate cancer tissue compared with the normal prostate tissue, which suggests that the gene expression is important to the arising of prostate cancer.

Amino Acid Sequence↗

PubMed2001

Molecular cloning and characterization of a novel peptidylprolyl isomerase (cyclophilin)-like gene (PPIL3) from human fetal brain.

During the large-scale sequencing analysis of a human fetal brain cDNA library, we isolated two cDNA clones encoding two novel proteins, which show 52% and 72% identity to the cyclophilin isoform 10 of C. elegans, respectively. Sequence analysis revealed these two cDNA clones are two different splicing variants of a novel cyclophilin-like gene (PPIL3). The PPIL3 gene was identified on a completely sequenced BAC (GenBank accession AC005037) from chromosome 2q33 between STS markers stSG2762 (proximal) and SHGC-3074 (distal), oriented toward the telomere. The PPIL3 gene consisted of eight exons spanning more than 18 kb of genomic DNA. RT-PCR analysis indicated that PPIL3 was ubiquitously expressed in adult human tissues.

5' Untranslated Regions↗

PubMed2001

[Study on the expression of pig zona pellucida-3 beta excluding N-terminus signal peptide and C-terminus transmembrance-like domain in Escherichia coli].

Pig oocytes are surrounded by an acellular translucent envelope-zona pellucida(pZP), which is comprised of three biochemically and immunologically distinct glycoproteins(pZP1, pZP3 alpha and pZP3 beta). Due to the cross-reactivity of anti-pZP3 beta antibodies with human ZP in vitro, pZP3 beta has been considered as potential one of immunogens for developing human contraceptive vaccine. In order to express pZP3 beta directly in E. coli; we amplified the core fragment of pZP3 beta cDNA by PCR that was deleted 5'- and 3'-terminal sequences of coding for signal peptide and transmembrane-like domain. The DNA sequenced EcoRI and SalI restriced core fragment was cloned in a frame downstream of PRPL promoter in the pBV221 vector. SDS-PAGE analysis showed that pZP3 beta protein was expressed especially in E. coli after thermal induction, which the expression band displayed the molecular weight of about 38 kD matching with its deduced molecular weight(36.5 kD). In addition, the specially expressed protein band on SDS-PAGE gel showed specific immunological reaction with anti-pig ZP IgGs of rabbit in Western blot.

Animals↗

PubMed1999

[Inhibitory effects of recombinant human neurotrophin-4/5 protein on neurotoxicity caused by arsenic trioxide].

OBJECTIVE: To study preliminarily the inhibitory effects of recombinant human neurotrophin (hNT)-4/5 protein on neurotoxicity caused by arsenic trioxide (As(2)O(3)). METHODS: Nerve cells of the chicken embryonic forebrain and PC12 cells were co-cultured with recombinant hNT-4/5 protein and As(2)O(3) (0 - 16 micromol/L) for 24 and 48 hours, respectively, to observe the survival of nerve cells and the outgrowth of PC12 cells, with clone expression and recombination techniques. RESULTS: There was significant difference in survival rate of nerve cells between the experimental and control groups (P < 0.01) after co-culture for 48 hours, and survival rate and number of neuron outgrowth increased with the concentration of hNT-4/5 protein. CONCLUSION: Recombinant hNT-4/5 protein could inhibit neurotoxicity caused by As(2)O(3), which provided a basis to find an anti-toxic factor in the environment by genetic engineering.

Animals↗