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Biomedical subjects

Y Tong

Publications and source records attributed to Y Tong.

At least 55 records · Page 3Linked to original sources
PubMed1999

[Establishment of a high germline competent C57BL/6J ES cell line].

Eleven ES cell lines from C57BL/6J mice were established, with the primary culture of mice embryos as feeder cells and 1 x 10(3) units Leukemia Inhibitory Factor (LIF) in the DMEM (high glucose) medium. The frequency of establishment was 9.6%. Five of these ES cell lines were demonstrated karyotypically normal with diploid composition of over 70%. They shared the characteristics of early embryonic cells: positive for alkaline phosphatase activity and oct4 gene expression. They also showed high potency to differentiate into wide types of cells in vivo. Following injection of the blastocysts, three of them showed abilities to give rise to chimeras and MESPU35 cell line was identified to have high efficiency of colonization into the germline. The clones of MESPU35 cells maintained the germline competence and the mutant clone also retained the high potency to participate into the development of embryos. MESPU35 cells can serve as a valuable vehicle for the production of mutant mice.

Animals↗
PubMed1999

The clinical and epidemiological significance of serum PreS1/anti-PreS1 in HBV infection.

OBJECTIVE: To investigate whether PreS1 antigen and/or anti-PreS1 antibody in serum might have some significance different from routine hepatitis B virus (HBV) markers in hepatitis B virus infection. METHODS: Forty-six acute hepatitis B patients and 377 chronic persistent hepatitis patients were examined to assess the clinical significance of serum PreS1/anti-PreS1. HBV family clustering analysis was carried out in a village in Shanghai suburb area, and case-control study on role of PreS1 in HBV spreading was undertaken in Tangshan, Hebei province. RESULTS: PreS1 in HBsAg positive sera from chronic persistent hepatitis patients correlated with HBV DNA (P < 0.05), while the presence of PreS2 in HBsAg positive sera was much more common than that of PreS1 and had no correlation with HBeAg or HBV DNA. Among 46 patients with acute hepatitis B, the appearance of anti-PreS1 antibody in serum correlated with the disappearing or decreasing of both PreS1 and ALT. In antiviral treatment, PreS1 antigen turned negative much earlier than PreS2 and HBsAg in acute hepatitis B patients' sera. Family clustering analysis suggested that people with PreS1 positive in serum were more infective than those with HBsAg positive alone (P < 0.01). The case-control study showed that chronic hepatitis B patients with PreS1 positive in serum played a much more important role in HBV spreading than those with PreS1 negative (P < 0.01) and asymptomatic HBsAg carriers (P < 0.01), and the odds ratios (OR) were 3.8 and 3.2, respectively. CONCLUSIONS: Serum PreS1 closely correlates with active virus replication, and PreS1/anti-PreS1 status indicates the outcome of acute hepatitis B. The serum PreS1/anti-PreS1 marker is of some significance in HBV epidemiology as well.

Biomarkers↗
PubMed1999

[Regulation of gene co-expression after bicistronic retrovirus-mediated gene transfer into human cord blood CD34+ cells].

OBJECTIVE: To compare internal SV40 early promoter with IRES of the in regulatory effect on gene co-expression after retrovirus-mediated gene transfer into human cord blood CD34+ cells. METHODS: Based on pLXSN, vectors pLESN and pLEIN carrying EGFP and neo gene, distinguished by the in sequences between two, genes, were constructed. After liposome-mediated gene transfer into packaging cells, high-titre virus supernatant was obtained. After supernatant infection in vitro, transduced cells were cultured in CFC assay. RESULTS: FACS analysis demonstrated that 10% of human cord blood CD34+ cell were infected in vitro. All of G418 resistant colonies demonstrated green fluorescence in LEIN group, while only 41.1% in LESN group. CONCLUSION: IRES is superior to SV40 promoter for the guarantee of co-expression in bicistronic retrovirus-mediated gene therapy in human cord blood CD34+ cells.

Adenoviruses, Human↗
PubMed1999

[Synergistic interaction between Ys-96, a bisbenzylisoquinoline compound derived from Stephania tetrandra, and adriamycin or vincristine against human cancer cell lines in vitro].

OBJECTIVE: To study in vitro the anticancer interaction between Ys-96, a bisbenzylisoquinoline compound derived from Stephania tetrandra, and adriamycin or vincristine against human cancer cell lines. METHOD: Using human breast cancer cell MCF-7 and its adriamycin-resistant cell line MCF-7/Adr, and human nasopharyngeal cancer cell KB and its vincristine-resistant cell line KBv200 in an in vitro system, anticancer interaction between Ys-96 and adriamycin or vincristine was evaluated with a method reported by Berenbaum. RESULT: The SFIC values (sum of fractional inhibitory concentration) of the combinations with 3 different ratios between Ys-96 and adriamycin or vincristine were markedly less than 1.0, and the shapes of all the isobologram curves were concave. CONCLUSION: The synergistic interaction between Ys-96 and adriamycin or vincristine against the above human cancer cell lines was positively observed in vitro.

Antineoplastic Agents, Phytogenic↗
PubMed1999

[Purification and partial characterization of antagonistic proteins from Bacillus subtilis B034].

Bacillus subtilis B034, an antagonistic bacteria isolated from the phyllosphere of rice, strongly inhibits the growth of Xanthomonas oryzae pv. oryzae which causes the rice bacterial blight. Crude extract was obtained by precipitation of the cell-free culture of B034 with 70% saturated (NH4)2SO4. The suspension of the precipitate strongly inhibits Xanthomonas oryzae pv. o-ryzae, and it is thermostable, resistant to trypsin, partial sensitive to proteinase K, pronase E and chloroform. Its active pH range is wide, from 4.0 to more than 12.0, relative more stable in high pH. Two antagonistic peaks were obtained from the crude extract of B034 after Phenyl-Sepharose CL-4B, DEAE-Sephacel and FPLC Superdex 75 HR 10/30 column chromatography. One of these two peaks, named P2, was showed only single band with 50.3 kD and pI6.25 in SDS-PAGE and PAGE-IEF, respectively. The N-terminal partial sequence of protein P2 was analyzed.

Amino Acid Sequence↗
PubMed1999

[Study of conformation of serum albumin by FTIR].

This paper used the FTIR method to study the spectra of bovine serum albumin (BSA) and its solution. The secondary structure and conformation was assigned by FTIR-deconvolution analysis. The result indicated that BSA secondary structure in solid is different from in aqueous solution. As the decrease of BSA concentration from 3.241 x 10(-1) to 4.032 X 10(-2) g X mL(-1), the amide I band components at 1 609.84, 1 633.85,1 653.69, 1 681.16 and 1 694.88 cm(-1) shifted to 1 608. 24, 1 638.36, 1 656.10, 1 674.87 and 1 690.78 cm(-1), respectively, the component at 1 621.50 cm(-1) unchanged.

Animals↗
PubMed1998

Cloning and characterization of a novel mammalian PP2C isozyme.

PP2C is a structurally diversified protein phosphatase family with a wide range of functions in cellular signal transduction. A novel PP2C subtype, designated PP2Cdelta, was identified from a rat cDNA clone, which encodes a protein of 392 amino acid residues. While PP2Cdelta shares approximately 30% sequence identity in its catalytic domain with the mammalian PP2C, it lacks a 90-residue carboxyl-terminal sequence conserved in mammalian PP2C. Northern blot analysis showed that PP2Cdelta is widely expressed in rat tissues. The transcription of the PP2Cdelta gene was activated in response to stress, such as the addition of ethanol to the culture medium or UV irradiation of cells. Recombinant PP2Cdelta purified from bacteria exhibited a potent Mn2+-dependent serine/threonine phosphatase activity. Unlike other members of the PP2C family, the activity of PP2Cdelta was inhibited, rather than stimulated, by Mg2+. Transfection with PP2Cdelta resulted in inhibition of cell growth, precluding generation of stable 293 or CHO transfectants. Using a modified tetracycline-regulated PP2Cdelta-GFP dicistronic expression cassette, it was revealed that overexpression of PP2Cdelta blocked cell cycle progression and arrested cells at early S phase, resulting in inhibition of DNA synthesis and leading to cell death. These results suggest that PP2Cdelta plays a role in regulation of cell cycle progression via dephosphorylation of its substrates whose appropriate phosphorylation states might be crucial for cell proliferation.

Amino Acid Sequence↗
PubMed1998

Probing the folate-binding site of human thymidylate synthase by site-directed mutagenesis. Generation of mutants that confer resistance to raltitrexed, Thymitaq, and BW1843U89.

Human thymidylate synthase (TS) contains three highly conserved residues Ile-108, Leu-221, and Phe-225 that have been suggested to be important for cofactor and antifolate binding. To elucidate the role of these residues and generate drug-resistant human TS mutants, 14 variants with multiple substitutions of these three hydrophobic residues were created by site-directed mutagenesis and transfected into mouse TS-negative cells for complementation assays and cytotoxicity studies, and the mutant proteins expressed and characterized. The I108A mutant confers resistance to raltitrexed and Thymitaq with respective IC50 values 54- and 80-fold greater than wild-type but less resistance to BW1843U89 (6-fold). The F225W mutant displays resistance to BW1843U89 (17-fold increase in IC50 values), but no resistance to raltitrexed and Thymitaq. It also confers 8-fold resistance to fluorodeoxyuridine. Both the kinetic characterization of the altered enzymes and formation of antifolate-resistant colonies in mouse bone marrow cells that express mutant TS are in accord with the IC50 values for cytotoxicity noted above. The human TS mutants (I108A and F225W), by virtue of their desirable properties, including good catalytic function and resistance to antifolate TS inhibitors, confirm the importance of amino acid residues Ile-108 and Phe-225 in the binding of folate and its analogues. These novel mutants may be useful for gene transfer experiments to protect hematopoietic progenitor cells from the toxic effects of these drugs.

Animals↗
PubMed1998

Discrete distribution of the neuropeptide Y Y5 receptor gene in the human brain: an in situ hybridization study.

The present study investigated the regional distribution of putative 'food-intake'-related neuropeptide Y Y5 receptor gene using cRNA in situ hybridization in various regions of the normal control post-mortem human brain. Interestingly, significant levels of Y5 receptor expression were detected in the hypothalamus; the arcuate nucleus being particularly enriched compared to other hypothalamic nuclei. Surprisingly, strong hybridization signals were also noted in the stratum granulosum of the dentate gyrus contrasting with lower levels of Y5 receptor transcripts in other regions of the hippocampal formation. The cerebral cortex, basal ganglia and thalamus were not enriched with Y5 receptor mRNA. It thus appears that the expression of the Y5 receptor gene in the human brain is rather restricted with enrichment in areas consistent with the involvement of this receptor type in the modulation of appetite and seizures.

Aged↗
PubMed1998

Conformational probes for elucidating the nature of substance P binding to the NK1 receptor: initial efforts to map the Phe7-Phe8 region.

Three substance P analogs with conformation constraints in the Phe7-Phe8 region have been prepared in connection with an effort to differentiate two families of potential conformations for the binding of substance P to its NK1 receptor. While the analogs did not bind the NK1 receptor with high affinity, the synthesis of the analogs demonstrated the utility of a general method for constructing piperazinone based peptidomimetics.

Amino Acid Sequence↗
PubMed1998

Isolation and characterization of thymitaq (AG337) and 5-fluoro-2-deoxyuridylate-resistant mutants of human thymidylate synthase from ethyl methanesulfonate-exposed human sarcoma HT1080 cells.

Thymidylate synthase plays an essential role in the synthesis of DNA. Recently, several new and specific thymidylate synthase inhibitors that occupy the folate binding site, including Tomudex(R), BW1843U89, and Thymitaq, have demonstrated therapeutic activity in patients with advanced cancer. In order to find drug-resistant forms of human thymidylate synthase for gene therapy applications, human sarcoma HT1080 cells were exposed to ethyl methanesulfonate and Thymitaq selection. Thymitaq-resistant clonal derived sublines were established, and analysis indicated that both gene amplification and point mutations contributed to drug resistance. Eight mutant cDNAs that were identified from Thymitaq-resistant sublines were generated by site-directed mutagenesis and transfected into thymidylate synthase-negative cells. Only K47E, D49G, or G52S mutants retain enzyme activity. Moreover, cytotoxicity studies demonstrated that D49G and G52S transfected cells, besides displaying resistance to Thymitaq with IC50 values 40- and 12-fold greater than wild-type enzyme transfected cells, respectively, also lead to fluorodeoxyuridine resistance (26- and 97-fold in IC50 values, respectively) but not to Tomudex or BW1843U89. Characterization of the purified altered enzymes obtained from expression in Escherichia coli is consistent with the cell growth inhibition results. We postulate that the D49G or G52S mutation leads to the structural perturbation of the highly conserved Arg50 loop, decreasing the binding of thymidylate synthase to the inhibitors, Thymitaq and fluorodeoxyuridylate.

Cloning, Molecular↗
PubMed1998

Regulation of expression of neuropeptide Y Y1 and Y2 receptors in the arcuate nucleus of fasted rats.

Neuropeptide Y is expressed in neurons of the hypothalamic arcuate nucleus and has been ascribed a role as a stimulant of food intake. Neuropeptide Y Y1 and Y2 receptors are also localised in the arcuate nucleus, and it has been suggested that the Y1 receptor mediates part of the effect of neuropeptide Y on feeding behaviour. In the present study, immunohistochemistry and in situ hybridization were used to investigate the effect of food deprivation on the expression of Y1 and Y2 receptors in the arcuate nucleus of the rat. Fasting for 48 h induced a decrease in the number and area of Y1 receptor immunoreactive neurons in the arcuate nucleus. Furthermore, arcuate Y1 receptor mRNA levels also decreased after food deprivation. The decrease in the number of the Y1 receptor immunoreactive neurons was partially attenuated by supplementing the drinking water with 10% glucose. In contrast, fasting did not significantly change Y2 receptor mRNA levels in the arcuate nucleus. These results support the view that Y1 receptors in the arcuate nucleus play a role in the feeding pattern induced by neuropeptide Y.

Animals↗
PubMed1998

Expression of Hras-p21 and keratin K13 in UVR-induced skin tumors in Sencar mice.

An ultraviolet radiation (UVR)-induced Sencar mouse skin carcinogenesis model was established to investigate the expression of Hras-p21 and keratin K13 in different stages of carcinogenesis, including UV-exposed nontumor skin, papillomas, squamous-cell carcinomas (SCCs), and malignant spindle-cell tumors (SCTs). Expression of Hras-p21 and K13 was examined in paraffin-embedded tumor sections by using immunohistochemical, immunofluorescent, and double staining techniques with specific antibodies. Positive Hras-p21 staining was detected in 1/3 (33%) papillomas, 24/36 (67%) of SCCs, but not in UVR-exposed nontumor skin or SCTs. Positive staining of the malignant progression marker K13 was found in 22/36 (61%) of SCCs only. Coexpression of Hras-p21 and K13 was found in 17/36(47%) SCCs. H-ras exons 1 and 2 were amplified from skin/tumor sections by using nested polymerase chain reaction (PCR). PCR-based single-strand conformation polymorphism (SSCP) analysis and gene sequencing revealed three point mutations, one in UVR-exposed nontumor skin (codon 56), and two in SCCs (codons 13 and 21). There were no clear relationships between point mutations of H-ras and the positive staining of Hras-p21 and K13. These results indicate that overexpression of ras-p21 in conjunction with aberrant expression of K13 is a frequent event in UVR-induced SCCs in Sencar mouse skin. Point mutation of the H-ras gene appeared to be a rare event in UVR skin carcinogenesis and not to be responsible for overexpression of Hras-p21.

Animals↗
PubMed1998

Should peri-operative infra-inguinal graft duplex scans be routinely performed?

BACKGROUND: The ability of duplex ultrasound graft surveillance to detect graft stenosis is well documented. However, the optimal time to commence duplex ultrasound graft surveillance is not clear. METHODS: An early duplex ultrasound study was performed on 90 infra-inguinal bypass grafts within 2 weeks of surgery. Further duplex scans were performed at intervals after surgery of 1 month, 3 months, 6 months, 1 year and then annually. An ankle/brachial systolic pressure index (ABI) was performed before the infra-inguinal bypass and then 24 h after the surgery. The ABI was also routinely measured at the time of each graft duplex examination. RESULTS: Six graft thromboses (6.7%) were demonstrated on the first graft duplex within 2 weeks of the surgery. The 24-hour postoperative resting ABI were less than 0.5 in all six cases. Significant graft stenoses (> 50%) or native artery diseases were detected in six cases (6.7%) on the first graft scan (proximal anastomosis stenosis, 1: mid-graft stenosis, 3; peroneal artery stenosis, 1; popliteal artery occlusion, 1). The resting ABI was less than 0.5 in one of these cases. Vein patch angioplasty was performed on graft stenosis in two cases following the first duplex scan. In 78 cases with either normal or minimal graft/native artery stenosis (< 50%), the resting ABI were less than 0.5 in only two cases (2.6%). The first graft duplex ultrasound examinations were technically difficult or gave a poor image in 21 cases (23.3%). During the subsequent graft duplex follow-up, seven graft thromboses were demonstrated at an average of 5.9 months after surgery (1-12 months). Significant stenosis (> 50%) was demonstrated in 10 additional cases at an average of 8.5 months after surgery (1-12 months). CONCLUSIONS: The data of the present study do not support routine peri-operative graft duplex scanning in addition to a 1-month graft duplex scan. Early postoperative resting ABI should be routinely performed. If the resting ABI is less than 0.5, an immediate duplex scan should be carried out to assess graft integrity. A long-term non-invasive graft surveillance programme is important in the detection of graft stenosis.

Aneurysm↗
PubMed1998

[Molecular cloning and DNA sequencing of OspC gene of two strains Borrelia burgdorferi isolated in China].

OBJECTIVE: To investigate the variation of OspC gene in two Chinese isolates of Borrelia burgdorferi. METHODS: PCR technique was used to amplify the OspC gene from the whole cellular DNA of isolates BT01 and BJ-9011. The amplified products were inserted into plasmid pGEM-3ZF(+) and sequenced. RESULTS: Except the signal peptide, the OspC genes of the two isolates BT01 and BJ-9011 were 579 bp and 576 bp which encode 193, 192 amino acids respectively. The nucleotide and amino acids sequence identity between the two strains was 86% and 83%. High homology exists between these Chinese isolates and several foreign isolates (PBi, PKo, B31), especially in BJ-9011. It had 99% nucleotide and amino acid sequence identified with B31. CONCLUSION: Variations in OspC genes are noted between the two Chinese Borrelia burgdorferi isolates and foreign isolates.

Antigens, Bacterial↗
PubMed1998

[Reversal effect of Ys-96, a bisbenzylisoquinoline, on adriamycin or vincristine resistance in human cancer cells in vitro].

OBJECTIVE: Reversal effect of Ys-96, a bisbenzylisoquinoline, on the resistance of human cancer cells to adriamycin or vincristine was studied in vitro. METHOD: In an in vitro culture system of human cancer cells MCF-7 and its adriamycin-resistant line MCF-7/Ad or KB and its vincristine-resistant line KBv200, the sensitivity(IC50) of the resistant cell lines to adriamycin or vincristine was evaluated with a MTT assay. RESULT: IC50 value of adriamycin or vincristine in combination with Ys-96 at a concentration of 1.00 mumol/L against MCF-7/Ad or KBv200 was found to be close to that of adriamycin alone or vincristine alone against the sensitive cell line MCF-7 or KB. CONCLUSION: The drug resistance of MCF-7/Ad or KBv200 could be essentially reversed by Ys-96 at a concentration of 1.00 mumol/L.

Benzylisoquinolines↗