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Biomedical subjects

Y Ke

Publications and source records attributed to Y Ke.

At least 55 records · Page 3Linked to original sources
PubMed2000

Multiple origins of Tibetan Y chromosomes.

The genetic origin of Tibetans was investigated using Y chromosome markers. A total of three populations were studied, two from central Tibet speaking central Tibetan and one from Yunnan speaking Kham. Two dominant paternal lineages (>80%) were identified in all three populations with one possibly from central Asia (YAP+) and the other from east Asia (M122C). We conclude that Tibetan Y chromosomes may have been derived from two different gene pools, given the virtual absence of M122C in central Asia and YAP+ in east Asia, with drift an unlikely mechanism accounting for these observations.

Alu Elements↗
PubMed2000

Basic fibroblast growth factor and angiogenesis in squamous carcinoma of the tongue.

The relationship between basic fibroblast growth factor (bFGF), receptors for bFGF and neoangiogenesis was investigated in 51 patients with squamous cell carcinoma of the tongue, 26 of whom had metastatic disease in cervical lymph nodes. Vessels were demonstrated by immunocytochemical labelling for CD34 and expressed as raw counts and volume-weighted counts. bFGF protein and its receptors FGFR1(flg) and FGFR2(bek), were demonstrated using immunocytochemical labelling. In situ hybridisation for bFGF mRNA was performed using a 250-bp digoxigenin-labelled RNA probe. In normal epithelium, the expression of bFGF protein and mRNA was more intense in the basal layer, while receptors for bFGF were expressed more strongly in the superficial parts. In carcinomas, expression of bFGF was greater in the more poorly-differentiated cells, but showed no relation to the overall tumour differentiation. There was strong bFGF expression in tumour-infiltrating lymphocytes. The expression of bFGF receptors was variable, with FGFR2 being particularly high in areas of keratinisation. There were no consistent changes in bFGF or receptor expression between primary carcinomas and their lymph node metastases, and there was no correlation with measures of vascularity or tumour growth pattern. bFGF is synthesised by all squamous carcinomas and has the potential to modulate angiogenesis. However, these data suggest that changes in the expression of bFGF and its receptors are not related to the intensity of neoangiogenesis in lingual carcinomas or their nodal metastases.

Carcinoma, Squamous Cell↗
PubMed2000

In vivo measurements of glutamine + glutamate (Glx) and N-acetyl aspartate (NAA) levels in human partial epilepsy.

OBJECTIVE: To investigate whether cerebral levels of N-acetyl aspartate (NAA), and glutamine + glutamate (Glx), are interictally altered in the epileptogenic regions of patients with partial seizures. MATERIAL AND METHODS: NAA, Glx, creatine (Cr), choline (Cho) and myo-inositol (mI) was measured in 28 patients with partial epilepsy and 10 healthy controls using localized 1H magnetic resonance spectroscopy. According to the multimethodological consensus, the epileptogenic region was mesial temporal in 18 and neocortical in 10 patients. RESULTS: The Glx/NAA and Glx/Cr ratios in epileptogenic regions were higher, and the NAA/Cr ratios lower than in the homologous regions (P=0.013, P=0.002 and P<0.0001). Applying the 95% confidence interval of controls, 17 of the 20 mesial temporal epileptogenic regions were correctly identified by an increased Glx/NAA and 15 of 20 by a decreased NAA/Cr ratio. Among patients with neocortical epilepsy the Glx/NAA ratio was increased in 8 of the 10 epileptogenic regions, whereas the NAA/Cr ratio was decreased in three. CONCLUSION: Both Glx and NAA are useful to identify the epileptogenic zone. The Glx/NAA ratios may be particularly useful to indentify neocortical epileptogenic regions.

Adolescent↗
PubMed2000

Transferrin-bound and transferrin free iron uptake by cultured rat astrocytes.

Previously we had demonstrated the presence of transferrin receptor (TfR) on the plasma membrane of cultured rat cortical astrocytes. In this study, we investigated the roles of TfR in transferrin-bound iron (Tf-Fe) as well as transferrin-free iron (Fe II) uptake by the cells. The cultured rat astrocytes were incubated with 1 microM of double-labelled transferrin (125I-Tf-59Fe) in serum- free DMEM F12 medium or 59Fe II in isotonic sucrose solution at 37 degrees C or 4 degrees C for varying times. The cellular Tf-Fe, Tf and Fe II uptake was analyzed by measuring the intracellular radioactivity with gamma counter. The result showed that Tf-Fe uptake kept increasing in a linear manner at least in the first 30-min. In contrast to Tf-Fe uptake, the internalization of Tf into the cells was rapid initially but then slowed to a plateau level after 10 min. of incubation. The addition of either NH4Cl or CH3NH2, the blockers of Tf-Fe uptake via inhibiting iron release from Tf within endosomes, decreased the cellular Tf-Fe uptake but had no significant effect on Tf uptake. Pre-treated cells with trypsin inhibited significantly the cellular uptake of Tf-Fe as well as Tf. These findings suggested that Tf-Fe transport across the membrane of astrocytes is mediated by Tf-TfR endocytosis. The results of transferrin-free iron uptake indicated that the cultured rat cortical astrocytes had the capacity to acquire Fe II. The highest uptake of Fe II occurred at pH 6.5. The Fe II uptake was time and temperature dependent, iron concentration saturable, inhibited by several divalent metal ions, such as Co2+, Zn2+, Mn2+ and Ni2+ and not significantly affected by phenylarsine oxide treatment. These characteristics of Fe II uptake by the cultured astrocytes suggested that Fe II uptake is not mediated by TfR and implied that a carrier-mediated iron transport system might be present on the membrane of the cultured cells.

Animals↗
PubMed2000

[Construction of BPI23-Fc gamma 1 recombinant protein prokaryotic expression vector and the expression and biological assessment of BPI23-Fc gamma 1 recombinant protein].

OBJECTIVE: To construct pBV-BPI600-Fc gamma 1(700) recombinant expression vector, to transform into Escherichia coli DH5 alpha, and to induce the expression of BPI23-Fc gamma 1 anti-bacterial recombinant protein. METHODS: Genes which encode BPI23 and Fc gamma 1 were amplified by RT-PCR from mRNA that was extracted from HL-60 cell and normal human leukocytes; Recombinant cloning vector and recombinant expression vector were constructed. pBV-BPI600-Fc gamma 1(700) recombinant expression vector was transformed into the competent Escherichia coli DH5 alpha and BPI23-Fc gamma 1 recombinant protein was expressed by temperature induced method. RESULTS: (1) Expected amplified products BPI600 bp and Fc gamma 1(700) bp were obtained by RT-PCR. (2) pUC18-BPI180, pUC18-BPI420, and pUC18-Fc gamma 1(700) recombinant cloning vector were successfully constructed, and sequences were identical with the reported ones. (3) pBV-BPI600-Fc gamma 1(700) recombinant expression vector was successfully constructed, and results of the enzyme digestion analysis were identical with expected ones. (4) pBV-BPI600-Fc gamma 1(700) recombinant expression vector was transformed into the competent Escherichia coli DH5 alpha and BPI23-Fc gamma 1 recombinant protein was expressed by temperature-induced method, and its expression level was accounted for 20% of total bacterial proteins. (5) The renatured BPI23-Fc gamma 1 recombinant protein had anti-bacterial activity and biological functions of complement fixation, opsonization. CONCLUSION: pBV-BPI600-Fc gamma 1(700) recombinant expression vector was successfully constructed, and BPI23-Fc gamma 1 recombinant protein with BPI and IgGFc double biological activity was expressed in Escherichia coli.

Antimicrobial Cationic Peptides↗
PubMed1999

Influence of the alpha1-adrenergic antagonist, doxazosin, on noradrenaline-induced modulation of cytoskeletal proteins in cultured hyperplastic prostatic stromal cells.

BACKGROUND: Doxazosin, an alpha1-adrenergic antagonist, inhibits sympathetic contraction of prostatic stromal smooth muscle cells and is used in the relief of obstructive benign prostatic hyperplasia (BPH). In vitro application of noradrenaline stimulates expression of cytoskeletal filaments, particularly actin and myosin, by prostatic stromal cells, thus enhancing their differentiation towards smooth muscle cells. This study examined the possible role of doxazosin in reversing this phenotypic modulation as well as in inhibiting smooth muscle cell contraction. METHODS: Stromal cell tissue cultures derived from 10 human hyperplastic prostates were rendered quiescent by reduction of stripped fetal calf serum (FCS) to 1% (v/v) in the medium followed by treatment with 20 microM noradrenaline and/or 1 microM doxazosin for 10 days. Doxazosin, in 10-fold increments of concentration, was also added, separately, to two of these cell cultures, which were either quiescent or growing in 10% normal (unstripped) FCS. Harvested cells were labelled with fluorescein-labelled antisera to smooth muscle cytoskeletal filaments, and their individual fluorescence levels were analyzed flow-cytometrically. RESULTS: Noradrenaline increased expression of all cytoskeletal filaments studied. This effect was greatest for actin and myosin in proliferating cell cultures. Doxazosin largely reversed the increase in filament expression. This effect was most significant for actin and myosin and greatest in quiescent cultures. However, inhibition of the agonist effect of noradrenaline by doxazosin showed no clear dose-related response, in that expression of cytoskeletal filaments was differentially inhibited. CONCLUSIONS: The data suggest that doxazosin may inhibit not only stromal contraction of differentiated smooth muscle cells in BPH but also the phenotypic modulation of stromal smooth muscle cell differentiation induced by noradrenaline. These actions, together, may render prostatic stroma less contractile, and hence less able to respond to sympathetic stimulation, in patients with BPH. While effects on isolated stromal cells are of undoubted importance, failure to demonstrate a consistent dose-response relationship between expression of smooth muscle cell phenotype and inhibition by doxazosin suggests that additional influences, including humoral factors as well as the proximity of differentiated epithelium, are also likely to be involved in this interaction in the intact tissue.

Adrenergic alpha-1 Receptor Antagonists↗
PubMed1999

Systematic comparison of gene expression through analysis of cDNA fragments within or near to the protein-coding region.

Life is controlled by the timely and ordered expression of genes. Identification of important genes involved in specific physiological and pathological conditions requires efficient methods to analyse differential gene expression. We describe a novel strategy, namely complete comparison of gene expression (CCGE), for a systematic assessment of differentially expressed genes. Using the CCGE method, double-stranded cDNA is digested with two restriction enzymes that cut with different frequencies, the representative cDNA fragments are generated within or near to the protein-coding region. After being flanked by two different types of adapters, and amplified by a nested suppression PCR, the selected cDNA fragments, representing entire cDNA population, can be divided into 256 subsets; amplified and compared in a systematic manner.

Animals↗
PubMed1999

Cerebellar granule cells acquire transferrin-free iron by a carrier-mediated process.

In this study, the mechanism of transferrin-free iron uptake by brain neuronal cells was investigated using the cultured cerebellar granule cells. Effects of incubation time, iron concentration, temperature and other divalent metals on the cellular uptake were determined. After five days of plating, the cells were incubated with different concentrations of transferrin-free iron in isotonic sucrose solution at different temperatures for a certain time. The cellular transferrin-free iron uptake was analysed by measuring the cellular radioactivity with a gamma-counter. The result showed that the cultured cerebellar granule cells had the capacity to acquire transferrin-free iron at pH 6.5, at which it was demonstrated that transferrin binds iron very poorly and only very little transferrin can be internalized by reticulocytes and HeLa cells. The iron uptake by cells increased with incubation time in a linear manner at a rate of 0.1076 pmol/microg protein/min within the first 10 min. The uptake was time- and temperature-dependent, iron concentration saturable, and inhibited by several divalent metal ions, such as Co2+, Zn2+, Mn2+ and Ni2+. These characteristics of transferrin-free iron uptake by the cultured cerebellar granule cells observed in this study, similar to those obtained from cells outside of the brain, implied that a carrier-mediated iron transport system might be present on the membrane of this type of brain neuronal cells. In addition, no significant difference in malondialdehyde measurement was found when the cells were incubated without or with the lower concentrations of iron (< 4 microM) for 20 min at 37 degrees C, demonstrating that this system was valid for studying membrane iron transport in this type of brain neuronal cell.

Animals↗
PubMed1999

[Study of APC, Rb, c-met gene copy numbers of human gastric mucosa epithelial cell line GES-1].

OBJECTIVE: To study the copy number of oncogene/tumor suppressor genes in GES-1 cell line to identify its characteristics. METHODS: Bio-14-dATP was incorporated into APC, Rb, c-met gene cloned in plasmid by nick translation. Fluorescence in situ hybridization (FISH) of interphase nuclei of GES-1 cells was performed. RESULTS: Two copies of APC were shown in 48% interphase nuclei and 3 copies in 22%; 71% and 80% cells had normal copies of Rb and c-met genes in their nuclei, respectively. CONCLUSION: GES-1 cell line is a relatively normal gastric mucosa epithelial cell line and can be used as a human in vitro model system for the study of carcinogenesis.

Cell Line↗
PubMed1999

[A study of airway reactivity and nature of chronic asthmatic bronchitis].

OBJECTIVE: To observe the airway reactivity with methacholine provocation and exercise challenge of chronic asthmatic bronchitis and to assess preliminarily the nature of chronic asthmatic bronchitis. METHODS: 42 patients with bronchial asthma, 29 with simple chronic bronchitis and 27 with chronic asthmatic bronchitis were selected. The study protocol consisted of inhalation challenge with methacholine and exercise challenge. Count of blood eosinophil cells and level of blood total IgE were studied. Serum eosinophil cationic protein (ECP) concentration was measured in the asthmatic bronchitis group. RESULTS: The positive ratio to methacholine response was 77.8% in chronic asthmatic bronchitis group, whereas the ratios were 100.0% and 41.4% in bronchial asthma and simple chronic bronchitis groups respectively. According to the response to exercise challenge, patients with chronic asthmatic bronchitis were divided into positive and negative groups. The cumulative dose of methacholine producing a 20% fall in FEV(1) (PD(20)-FEV(1)) was different between the two groups. In the positive group (9 patients), none was negative with methacholine response and PD(20)-FEV(1) being similar to the bronchial asthma group. The blood total IgE and ECP levels were significantly higher in the positive than in the negative group. CONCLUSION: The characteristics of airway reactivity in chronic asthmatic bronchitis are midway between bronchial asthma and simple chronic bronchitis. The nature of chronic asthmatic bronchitis may be different in different patients.

Adult↗
PubMed1999

[Studies on the antitumour effect of Alocasia macrorrhiza].

Models of transplanted tumour in mice and human cancer enograft in nude mice were used to evaluate the antitumour effect of water extract of Alocasia macrorrhiza. Results showed that the inhibitory rate against S180 in mice was 29.38%, and the inhibitory rate against transplantable humman gastroadenitis in nude mice was 51.72%. No antitumour effect was shown against ECA in mice.

Adenocarcinoma↗
PubMed1998

[The use of microsatellite DNA markers for distinguishing metastatic tumor cells].

OBJECTIVE: To study the genetic stability and to establish a method for detection of micrometastasis using microsatellite DNA in human breast cancer xenograft in nude mice. METHODS: Fresh tissue of human breast cancer was xenotransplanted in nude mice or thotopically. Genomic DNA extracted from tissues of human breast cancer, xenotransplanted tumors and metastatic foci in nude mice were PCR amplified at three microsatellite loci (D14S68, D18S69, D20S199) and were analysed by electrophoresis and silver stain on PAGE. RESULTS Microsatellite DNA in genome of the xenotransplanted tumors and metastatic foci in nude mice were identical with that of the human breast cancer. CONCLUSION: This study has demonstrated in nude mice the xenotransplanted tumors and metastatic foci that originated from human breast cancer. The genetic stability in human breast cancer is evident in the processes of xenotransplantation, serial passages in nude mice, metastasis and in vitro culture. This method is sensitive and specific for the discrimination of metastatic tumor cells.

Animals↗
PubMed1998

Loops and bulge/loops in iron-responsive element isoforms influence iron regulatory protein binding. Fine-tuning of mRNA regulation?

A family of noncoding mRNA sequences, iron-responsive elements (IREs), coordinately regulate several mRNAs through binding a family of mRNA-specific proteins, iron regulatory proteins (IRPs). IREs are hairpins with a constant terminal loop and base-paired stems interrupted by an internal loop/bulge (in ferritin mRNA) or a C-bulge (in m-aconitase, erythroid aminolevulinate synthase, and transferrin receptor mRNAs). IRP2 binding requires the conserved C-G base pair in the terminal loop, whereas IRP1 binding occurs with the C-G or engineered U-A. Here we show the contribution of the IRE internal loop/bulge to IRP2 binding by comparing natural and engineered IRE variants. Conversion of the internal loop/bulge in the ferritin-IRE to a C-bulge, by deletion of U, decreased IRP2 binding by >95%, whereas IRP1 binding changed only 13%. Moreover, IRP2 binding to natural IREs with the C-bulge was similar to the DeltaU6 ferritin-IRE: >90% lower than the ferritin-IRE. The results predict mRNA-specific variation in IRE-dependent regulation in vivo and may relate to previously observed differences in iron-induced ferritin and m-aconitase synthesis in liver and cultured cells. Variations in IRE structure and cellular IRP1/IRP2 ratios can provide a range of finely tuned, mRNA-specific responses to the same (iron) signal.

5-Aminolevulinate Synthetase↗
PubMed1998

[Studies of microsatellite instability in Chinese gastric cancer tissues].

OBJECTIVE: To identify the microsatellite instability(MSI) rates in Chinese gastric cancer samples. METHODS: 29 microsatellite markers were selected to examine 42 paired gastric cancer tissues for MSI on all chromosomes except Y. RESULTS: The total frequency of MSI in all 42 gastric cancers was 33.9% with higher rates at loci of D3S1577, D3S1067,D8S279,D9S257, D1S248, D7S520 and D2S147,and the highest rate at D3S1577 and D3S1067(51.35%). MSI varied with different pathological types. The frequencies of MSI were signi- ficantly higher in poorly differentiated tumors and signet cell types, compared with well differentiated tumors(P=0.0026 and 0.0013 by chi-square test),and no difference was noted between poorly differentiated and signet cell types. CONCLUSION: MSI may play an important role in Chinese gastric cancer, particularly the poorly differentiated adenocarcinomas. The data presented here further support the previous hypothesis that pathologically distinct subtypes of gastric cancer undergo different genetic pathways during tumorigenesis.

Adult↗
PubMed1998

hSmad5 gene, a human hSmad family member: its full length cDNA, genomic structure, promoter region and mutation analysis in human tumors.

hSmad (mothers against decapentaplegic)-related proteins are important messengers within the Transforming Growth Factor-beta1 (TGF-beta1) superfamily signal transduction pathways. To further characterize a member of this family, we obtained a full length cDNA of the human hSmad5 (hSmad5) gene by rapid amplification of cDNA ends (RACE) and then determined the genomic structure of the gene. There are eight exons and two alternative transcripts; the shorter transcript lacks exon 2. We identified the hSmad5 promoter region from a human genomic YAC clone by obtaining the nucleotide sequence extending 1235 base pairs upstream of the 5' end of the cDNA. We found a CpG island consistent with a promoter region, and we demonstrated promoter activity in a 1232 bp fragment located upstream of the transcription initiation site. To investigate the frequency of somatic hSmad5 mutations in human cancers, we designed intron-based primers to examine coding regions by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analysis. Neither homozygous deletions or point mutations were found in 40 primary gastric tumors and 51 cell lines derived from diverse types of human cancer including 20 cell lines resistant to the growth inhibitory effects of TGF-beta1. These results suggest that the hSmad5 gene is not commonly mutated and that other genetic alterations mediate the loss of TGF-beta1 responsiveness in human cancers.

Alternative Splicing↗