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Biomedical subjects

Y Zeng

Publications and source records attributed to Y Zeng.

At least 145 records · Page 8Linked to original sources
PubMed1997

[Recombinant AAV-LMP-induced LMP specific cytotoxic response to autologous lymphoblastoid cell lines tranformed by Epstein-Barr virus].

Epstein-Barr virus is believed to be controlled in normal host by virus specific cytotoxic T lymphocytes (CTL). Although unable to eliminate EBV from the body, CTL seems to be essential in control of latently infected cells. Infusion of autologous EBV specific CTL, which can be produced in laboratory by separating lymphocytes from patients and stimulating them with EBV antigen, will provide an effective method of preventing and treating EBV-related diseases. We inserted the LMP gene of EB virus into an AAV vector pACP and packed it in Ad2 infected 293 cells by co-transfecting with plasmid Ad8, which produced the recombinant virus rAAV-LMP. The recombinant virus was used to infect stimulating cells and LMP antigen was expressed on the surface of these cells. Then the stimulating cells were irradiated and co-cultured with T lymphocytes. The EBV specific CTLs were obtained. The target cells were autologous LCLs from EBV-transformed B lymphocytes. The CTL activity was assayed by BLT activity method. The result indicated that all the four CTL strains could recognize and kill their target cells. This study has laid the technical basis for us to prevent and treat nasopharyngeal carcinoma in China with molecular biological methods.

Cell Line↗
PubMed1997

[Expression of Epstein-Barr virus latent membrane protein (EBV-LMP) in human laryngeal carcinoma].

In order to investigate the expression of Epstein-Barr virus in human laryngeal carcinoma, immunohistochemical study for Epstein-Barr virus latent membrane protein (EBV LMP) was performed on 90 laryngeal specimens taken from laryngeal carcinoma patients. The results showed that LMP was localized on the cell membrane and in cytoplasm. LMP was detected in 41 of 90 (45.5%) laryngeal squamous cell carcinoma, among them the positive rates of poorly differentiated, middle differentiated and well differentiated squamous carcinoma were 44% (12/27), 52% (25/47) and 26.6% (4/15) respectively. This study indicated that EB virus was not only presented in poorly differentiated carcinoma, it was also presented in well differentiated carcinoma. The development of laryngeal squamous carcinoma is related to the infection of EB virus.

Adult↗
PubMed1997

[Subtype and sequence analysis of the C2-V3 region of gp120 genes among human immunodeficiency virus infected IDUs in Ruili epidemic area of Yunnan Province of China].

DNA fragments of HIV-1 env gene were amplified by nested-PCR from 17 uncultured peripheral blood mononuclear cells (PBMCs) obtained from HIV-1 seropositive intravenous drug users (IDUs) in Ruili city of Yunnan Province. The C2-V3 region (about 450 bp) of them were sequenced. Sequence analysis showed that there exists two HIV-1 subtypes, B and C, with 5.8% and 2.2% gene divergence inside each subtype. The 12 subtype B strains, were closely related to those found in Thailand, Myanma and Ruili city of Yunnan, and the nucleotide sequence divergence between them ranged from 4.4% to 4.9%; meanwhile, the 5 subtype C strains were most close to those found in India as well as Ruili city, all with a genetic distance of 1.9%. The small divergence among Ruili HIV-1 subtype C strains suggests a recent epidemic. The analysis of V3 loop amino sequence of 12 subtype B HIV-1 reveals that V3-tip motif of 6 samples (50%) is GPGQ and that of 3 samples (25%) is GPGR. In addition, the codon of arginine (R) of all the strains is CGA instead of AGA. This result is in accordance with our previous hypothesis that there is a drift in vivo from GPGR to GPGQ motif on the tipof V3-loop of HIV-1 subtype B strain in this arm with the elapse of time.

Amino Acid Sequence↗
PubMed1997

[A seroepidemiological survey: antibody to HTLV-1 in sera from various populations in Guangdong Province].

A seroepidemiological survey of HTLV-1 infection in Guangdong Province was reported. 2224 serum samples from various populations were collected and antibodies to HTLV-1 in sera were detected with indirect immunofluorescent assay. Total seropositive rate was 1.57% (35/2224). The antibody positive rates of HTLV-1 in sera from healthy individuals (n = 1810), blood donors (n = 248), patients with T cell leukemia (n = 109) and patients with neurological diseases (n = 57) were 1.27%, 0.40 %, 7.30% and 5.26% respectively. There was a significant difference between the patients with T cell leukemia and the healthy individuals (P<0.005).

Antibodies, Viral↗
PubMed1997

[Surface-enhanced Raman spectra studies on four complexes derived from D-glucosamine and alpha-glycine and their effect on DNA].

Raman scattering of four complexes derived from d-glucosamine and alpha-glycine and their effects on DNA were studied with surface-enhanced Raman spectra (SERS). It was found that their SERS had shown little difference for their similar structural. Their interaction abilities on DNA were poor according to the change of their SERS spectrun after adding DNA to their water solution except for Co(III)GluG, which has strong effect on DNA. NiGluG's adsorptive ability on Ag sol is the strongest among other three complexes.

DNA↗
PubMed1996

Transcriptional repression by YY1 is mediated by interaction with a mammalian homolog of the yeast global regulator RPD3.

YY1 is a mammalian zinc-finger transcription factor with unusual structural and functional features. It has been implicated as a positive and a negative regulatory factor that binds to the CCATNTT consensus DNA element located in promoters of many cellular and viral genes. A mammalian cDNA that encodes a YY1-binding protein and possesses sequence homology with the yeast transcriptional factor RPD3 has been identified. A Gal4 DNA binding domain-mammalian RPD3 fusion protein strongly represses transcription from a promoter containing Gal4 binding sites. Association between YY1 and mammalian RPD3 requires a glycine-rich region on YY1. Mutations in this region abolish the interaction with mammalian RPD3 and eliminate transcriptional repression by YY1. These data suggest that YY1 negatively regulates transcription by tethering RPD3 to DNA as a cofactor and that this transcriptional mechanism is highly conserved from yeast to human.

Amino Acid Sequence↗
PubMed1996

Humoral response after administration of E1-deleted adenoviruses: immune privilege of the subretinal space.

An important limitation of E1-deleted recombinant adenoviruses in gene therapy is the immune response that they engender and that rapidly destroys transduced cells. Transduced cells of the outer retina appear to be an exception. To determine whether differences in immune sequestration of the outer retina contribute to the increased stability of transgene expression in this tissue, we examined the systemic humoral response to an E1-deleted adenovirus injected into the subretinal space. Subretinal injection of Ad.CMVlacZ in mature immunocompetent mice resulted in minimal circulating antibody production. In contrast, subcutaneous administration of equivalent doses of Ad.CMVlacZ resulted in high-titer antibody production against adenoviral proteins. Circulating antibodies in systemically immunized animals had minimal effect on retinal transgene expression patterns measured 2 weeks after subretinal injection of Ad.CMVlacZ. Histologic examination showed minimal retinal toxicity attributable to subretinal adenovirus in naive or immunized mice, although 3/18 (14%) of eyes from the latter set contained a localized granulomatous infiltrate at the ocular injection site. The data demonstrate that the subretinal space is an immune-privileged site regarding humoral immunity. Further, short-term transduction efficiency is not affected by the presence of anti-adenoviral antibodies. The retina may be a favorable environment for replication-defective virus-mediated genetic therapy.

Adenovirus E1 Proteins↗
PubMed1996

Synthesis of aryl azide derivatives of UDP-GlcNAc and UDP-GalNAc and their use for the affinity labeling of glycosyltransferases and the UDP-HexNAc pyrophosphorylase.

The chemical synthesis and utilization of two photoaffinity analogs, 125I-labeled 5-[3-(p-azidosalicylamido)-1-propenyl]-UDP-GlcNAc and -UDP-GalNAc, is described. Starting with either UDP-GlcNAc or UDP-GalNAc, the synthesis involved the preparation of the 5-mercuri-UDP-HexNAc and then attachment of an allylamine to the 5 position to give 5-(3-amino)allyl-UDP-HexNAc. This was followed by acylation with N-hydroxysuccinimide p-aminosalicylic acid to form the final product, i.e., 5-[3-(p-azidosalicylamido)-1-propenyl]-UDP-GlcNAc or UDP-GalNAc. These products could then be iodinated with chloramine T to give the 125I-derivatives. Both the UDP-GlcNAc and the UDP-GalNAc derivatives reacted in a concentration-dependent manner with a highly purified UDP-HexNAc pyrophosphorylase, and both specifically labeled the subunit(s) of this protein. The labeling of the protein by the UDP-GlcNAc derivative was inhibited in dose-dependent fashion by either unlabeled UDP-GlcNAc or unlabeled UDP-GalNAc. Likewise, labeling with the UDP-GalNAc probe was blocked by either UDP-GlcNAc or UDP-GalNAc. The UDP-GlcNAc probe also specifically labeled a partially purified preparation of GlcNAc transferase I.

Affinity Labels↗
PubMed1996

Inactivation of the mouse Brca1 gene leads to failure in the morphogenesis of the egg cylinder in early postimplantation development.

BRCA1 is proposed to be a tumor suppressor gene. To explore the biological function of BRCA1, a partial deletion (amino acids 300-361) of mouse Brca1 exon 11 was introduced into the genome of embryonic stem (ES) cells by homologous recombination. Mice carrying one mutated allele of Brca1 appear normal and are fertile up to 10 months of age without any sign of illness. However, no viable progeny homozygous for the Brca1 mutant allele were obtained. Detailed analysis of large numbers of embryos at different stages of development indicated that the homozygous mutant concepti are severely retarded in growth as early as embryonic day 4.5 (E4.5) and are resorbed completely by E8.5. Although the homozygotes at E5.5-E6.5 are able to synthesize DNA and display distinguishable embryonic and extraembryonic structures, they fail to differentiate and form egg cylinders. Consequently, they were unable to form primitive streaks and undergo gastrulation. Consistent with these in vivo results, blastocysts homozygous for mutated Brca1 alleles are at a considerable disadvantage when grown in vitro. These observations suggest that Brca1 has an important role in the early development of mouse embryos.

Animals↗
PubMed1996

Purification to homogeneity and properties of UDP-GlcNAc (GalNAc) pyrophosphorylase.

The pyrophosphorylase that condenses UTP and GlcNAc-1-P was purified 9500-fold to near homogeneity from the soluble fraction of pig liver extracts. At the final stage of purification, the enzyme was quite stable and could be kept for at least 4 months in the freezer with only slight loss of activity. On native gels, the purified enzyme showed a single protein band, and this band was estimated to have a molecular mass of approximately125 kDa on Sephacryl S-300. SDS-polyacrylamide gel electrophoresis analysis of the enzyme gave three protein bands of 64, 57, and 49 kDa, but these polypeptides are all closely related based on the following. 1) All three polypeptides show strong cross-reactivity with antibody prepared against the 64-kDa band. 2) All three proteins become labeled with either the UDP-GlcNAc photoaffinity probe azido-125I-salicylate-allylamine-UDP-GlcNAc or a similar UDP-GalNAc photoaffinity probe, and either labeling was inhibited in a specific and concentration-dependent manner by unlabeled UDP-GlcNAc or UDP-GalNAc. Thus, the enzyme is probably a homodimer composed of two 64-kDa subunits. The purified enzyme had an unusual specificity in that, at higher substrate concentrations, it utilized UDP-GalNAc as a substrate as well as UDP-GlcNAc in the reverse direction and GalNAc-1-P as well as GlcNAc-1-P in the forward direction. However, the Km for the GalNAc substrates was considerably higher than that for GlcNAc derivatives. This activity for synthesizing UDP-GalNAc was not due to epimerase activity since no UDP-GalNAc could be detected when the enzyme was incubated with UDP-GlcNAc for various periods of time. The pyrophosphorylase required a divalent cation, with Mn2+ being best at 0.5-1 mM, and the pH optimum was between 8.5 and 8.9.

Affinity Labels↗
PubMed1996

pH regulation in mouse sperm: identification of Na(+)-, Cl(-)-, and HCO3(-)-dependent and arylaminobenzoate-dependent regulatory mechanisms and characterization of their roles in sperm capacitation.

Intracellular pH (pHi) regulates several aspects of mammalian sperm function, although the transport mechanisms that control pHi in these cells are not understood. The pHi of mouse cauda epididymal sperm was determined from the fluorescence excitation ratio of 2,7-bis(carboxyethyl)-5(6)-carboxyfluorescein and calibrated with nigericin and elevated external [K+]. Two acid efflux mechanisms were identified following imposition of acid loads. One pathway has many anticipated characteristics of the somatic Na(+)-dependent Cl(-)-HCO3- exchanger, although sperm and somatic mechanisms can be distinguished by their ion selectivity and inhibitor sensitivity. Sperm may have an isoform of this exchange pathway with novel functional characteristics. The second acid-export pathway does not require extracellular anions or cations and is inhibited by arylaminobenzoates (flufenamic acid, diphenylamine-2-carboxylate). Mouse sperm also recover spontaneously from intracellular alkalinization. Recovery rates in N-methyl-D-glucamine+ Cl- or in 0.25 M sucrose are not significantly different from that in a complex culture medium. Thus, recovery from alkalinization does not utilize specific, ion-dependent transport mechanisms. Other widely distributed acid-efflux mechanisms, such as the Na(+)-H+ antiport pathway and the Na(+)-independent Cl(-)-HCO3- exchanger are not major regulators of mouse sperm pHi. Sperm capacitation results in pHi increases (from 6.54 +/- 0.08 to 6.73 +/- 0.09) that require a functional Na(+)-, Cl(-)-, and HCO3(-)-dependent acid-efflux pathway. Inhibition of this regulatory mechanism attenuates alkaline shifts in pHi during capacitation as well as the ability of sperm to produce a secretory response to zona pellucida agonists. These data suggest that one aspect of mouse sperm capacitation is the selective activation of one major pHi regulator.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1996

Detection of Epstein-Barr Virus DNA in well and poorly differentiated nasopharyngeal carcinoma cell lines.

Undifferentiated and poorly differentiated nasopharyngeal carcinoma (NPC) were known to be tightly associated with Epstein-Barr Virus (EBV). Its association with well differentiated NPC was also reported. In the present study, the presence of EBV was investigated by nucleic acid hybridization, Polymerase Chain Reaction (PCR), Immunoblot and in situ hybridization in two well differentiated NPC cell lines (CNE-1 and HK-1) and two other poorly differentiated NPC cell line (CNE-2 and CNE-3). Contrary to previous report indicating the absence of EBV in these cell lines, EBV DNA and proteins were present in all cell lines. The detection of EBV became more easily when the investigation was carried out on the nude mice tumor induced by transplantation of each NPC epithelial cell line. The EBV latent membrane protein (LMP1) was found by in situ hybridization to be integrated partly in the chromosomal DNA of these cell lines. The observations indicate that EBV could persist for a long time in the carcinoma cells established directly from well and poorly differentiated tumor biopsies and from transplantable NPC tumor in nude mice.

Blotting, Southern↗
PubMed1996

Photoreceptor cell rescue in retinal degeneration (rd) mice by in vivo gene therapy.

Mutations in the beta subunit of the cGMP phosphodiesterase gene (beta PDE) can cause a recessively inherited retinal degeneration in several species, including mice, dogs and humans. We tested the possibility of altering the course of retinal degeneration in the rd mouse through subretinal injection of a recombinant replication-defective adenovirus that contains the murine cDNA for wild-type (beta PDE, Ad.CMV beta PDE. Subretinal injection of Ad.CMV beta PDE results in beta PDE transcripts and increased PDE activity and delays photoreceptor cell death by six weeks. The findings demonstrate cell rescue by in vivo gene transfer, thus supporting the feasibility of treating an inherited retinal degeneration by somatic gene therapy.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1996

Spousal-residence separation among Chinese young couples.

The factors affecting the residential separation of spouses in China are examined. "Based on the microdata of the 1987 National Population Survey, we find that the variation in spousal-residence separations among Chinese young couples in the mid-1980s is well explained by personal and household factors within a multivariate model. The separations were aggravated by migrations for the reasons of employment or education. Although marriage migrations reduced the number of separations, those who had been married for a short period of time...were more prone to be separated. It is ironic that the higher a person's level of education, the greater the tendency for them to suffer the pain of spousal-residence separation. Household status could also be a very important factor: the lower the household status of a married individual, the more likely that he (or she) would be separated from their spouse."

Asia↗
PubMed1996

ZP3-dependent activation of sperm cation channels regulates acrosomal secretion during mammalian fertilization.

The sperm acrosome reaction is a Ca(2+)-dependent secretory event required for fertilization. Adhesion to the egg's zona pellucida promotes Ca2+ influx through voltage-sensitive channels, thereby initiating secretion. We used potentiometric fluorescent probes to determine the role of sperm membrane potential in regulating Ca2+ entry. ZP3, the glycoprotein agonist of the zona pellucida, depolarizes sperm membranes by activating a pertussis toxin-insensitive mechanism with the characteristics of a poorly selective cation channel. ZP3 also activates a pertussis toxin-sensitive pathway that produces a transient rise in internal pH. The concerted effects of depolarization and alkalinization open voltage-sensitive Ca2+ channels. These observations suggest that mammalian sperm utilize membrane potential-dependent signal transduction mechanisms and that a depolarization pathway is an upstream transducing element coupling adhesion to secretion during fertilization.

Acrosome↗