Biomedical subjects

Z Gu

Publications and source records attributed to Z Gu.

At least 163 records · Page 9Linked to original sources

PubMed1995

Inhibition of human immunodeficiency virus type 1 multiplication by transforming growth factor beta 1 and AZT in HIV-1-infected myeloid cells.

Myeloid cells are important reservoirs of HIV-1 infection. In response to pathogenic agents, macrophages secrete inflammatory cytokines that can modulate viral replication and contribute to AIDS pathogenesis. Because HIV replication is dependent on cellular activation, immunosuppressive cytokines that deactivate macrophages and T cells may be important modulators of an antiviral effect. We tested the anti-HIV potential of the immunosuppressive cytokine-transforming growth factor beta (TGF-beta 1) alone and in combination with AZT in a new monomyeloblastic model of HIV-1 infection. The PLB-985 cell model was infected with HIV IIIB strain, and the course of HIV-1 infection and replication was monitored by reverse transcriptase assay, p24 immunofluorescence, and northern blot analysis of HIV-1-specific mRNA. TGF-beta 1 as a single agent had no effect on the multiplication of HIV-IIIB in de novo-infected PLB 985 monomyeloblastic cells. However, cotreatment with TGF-beta 1 and AZT synergistically slowed virus multiplication within the first week following infection, as determined by reverse transcriptase measurement, p24 antigen detection, and northern blot analysis of viral RNA. The synergistic actions of TGF-beta 1 and AZT were also observed in PLB 985 cells infected with an AZT-resistant strain of HIV-1 (HIV 1393). Synergism between nucleoside analogs and cytokines may be an important therapeutic approach to HIV-1 infection. Elucidation of the role of cytokines in controlling the degree of HIV multiplication may have an impact on both clinical treatments and understanding the progression to AIDS.

Acquired Immunodeficiency Syndrome↗

PubMed1995

Possible involvement of cell fusion and viral recombination in generation of human immunodeficiency virus variants that display dual resistance to AZT and 3TC.

We have attempted to relate genetic recombination involving human immunodeficiency virus type 1 (HIV-1) to multiple drug resistance by using PEG to fuse subclones of U937 cells that carried HIV-1 recombinants resistant to either 3' -azido-3'-deoxythymidine (AZT) or the (--) enantiomer of 2',3'-dideoxy-3'-thiacytidine (3TC). The parental viruses employed contained well-defined mutations in the pol gene. Fused cells were co-cultured with the MT4 lymphocyte cell line for virus amplification to yield progeny that, in some cases, possessed different patterns of drug resistance from parental viruses. Mutational analyses were performed by PCR to substantiate these observations, which were also confirmed by direct sequencing of single strands of DNA segments, obtained from plaque-purified viruses. These studies indicate that viral recombination had occurred, and establish a theoretical basis on which to conclude that the acquisition of multiple drug resistance on the part of HIV-1 may be related to its ability to promote cell fusion.

Antiviral Agents↗

PubMed1995

K65R mutation of human immunodeficiency virus type 1 reverse transcriptase encodes cross-resistance to 9-(2-phosphonylmethoxyethyl)adenine.

Cloned variants of human immunodeficiency virus type 1 that contain the K65R mutation in reverse transcriptase have previously been shown to display approximately 10- to 30-fold resistance against 2',3'-dideoxycytidine, 2',3'-dideoxyinosine, and 2',3'-dideoxy-3'-thiacytidine. On the basis of tissue culture studies with both primary T cells and established cell lines, we now report that the K65R mutation confers approximately 12- to 15-fold resistance to 9-(2-phosphonylmethoxyethyl)adenine (PMEA). Likewise, a chain termination system revealed that mutated recombinant K65R reverse transcriptase displays resistance to PMEA diphosphate, the active metabolite of PMEA, in cell-free enzyme assays. Parallel studies have shown that the M184V mutation in reverse transcriptase, associated with high-level resistance against the (-) enantiomer of 2',3'-dideoxy-3'-thiacytidine, does not confer resistance to PMEA in tissue culture. Viruses and enzymes that included both the K65R and M184V mutations were resistant to PMEA and PMEa diphosphate, respectively, but only to the extent conferred by the K65R mutation alone.

Adenine↗

PubMed1995

A gratuitous inducer of cat-86, amicetin, inhibits bacterial peptidyl transferase.

Expression of the chloramphenicol resistance gene cat-86 is regulated by translation attenuation. Among the three ribosomally targeted antibiotics that can induce the gene, only amicetin has an unknown mode of action. Here we demonstrate that the nucleoside antibiotic amicetin is an inhibitor of bacterial peptidyl transferase. Thus, the three inducers of cat-86, chloramphenicol, erythromycin, and amicetin, interact with the peptidyl transferase region of bacterial ribosomes.

Anti-Bacterial Agents↗

PubMed1995

NF-Y controls transcription of the minute virus of mice P4 promoter through interaction with an unusual binding site.

Electrophoretic mobility shift assays performed with nuclear extracts from human fibroblasts revealed the formation of two major protein complexes with an oligonucleotide (nucleotides 78 to 107) from the palindromic region located upstream from the minute virus of mice (MVM) P4 promoter. It was shown that this oligonucleotide bound USF at the enhancer E box CACATG. The second complex contained the transcription factor NF-Y, whose association was surprising because its target sequence lacks the canonical CCAAT motif present in all mammalian NF-Y binding sites identified so far. The MVM NF-Y recognition element instead contains the CCAAC sequence. USF and NF-Y had distinct but overlapping sequence requirements for binding, suggesting that their associations with MVM DNA were mutually exclusive. Because of the palindromic nature of MVM DNA terminal sequences, NF-Y associated with the three nucleotide configurations corresponding to the hairpin structure and to the external and internal arms of the extended duplex replication form, respectively. However, owing to the imperfection of the palindrome, the binding of USF was restricted to the internal arm. Point mutations that suppressed the in vitro binding of NF-Y to the internal palindromic arm reduced the activity of the resident P4 promoter, while those preventing complex formation with USF did not, as determined by transient expression assays using the luciferase reporter gene. The data led to the identification of a novel P4 upstream regulatory region capable of interacting with two transcription factors, from which one (NF-Y) appeared to upmodulate the activity of the promoter.

Base Sequence↗

PubMed1995

Effect of human papillomavirus type 16 oncogenes on MAP kinase activity.

The mitogen-activated protein (MAP) kinase signal transduction pathway is an intracellular signaling cascade which mediates cellular responses to growth and differentiation factors. The MAP kinase pathway can be activated by a wide range of stimuli dependent on the cell types, and this is normally a transient response. Oncogenes such as ras, src, raf, and mos have been proposed to transform cells in part by prolonging the activated stage of components within this signaling pathway. The human papillomavirus (HPV) oncogenes E6 and E7 play an essential role in the in vitro transformation of primary human keratinocytes and rodent cells. The HPV type 16 E5 gene has also been shown to have weak transforming activity and may enhance the epidermal growth factor (EGF)-mediated signal transduction to the nucleus. In the present study, we have investigated the effects of the oncogenic HPV type 16 E5, E6, and E7 genes on the induction of the MAP kinase signaling pathway. The E5 gene induced an increase in the MAP kinase activity both in the absence and in the presence of EGF. In comparison, the E6 and E7 oncoproteins do not alter the MAP kinase activity or prolong the MAP kinase activity induced with EGF. These findings suggest that E5 may function, at least in part, to enhance the cell response through the MAP kinase pathway. However, the transforming activity of E6 and E7 is not associated with alterations in the MAP kinase pathway. These findings are consistent with E5 enhancing the response to growth factor stimulation.

Animals↗

PubMed1995

[Molecular basis and clinical significance of HIV-1 resistance to nucleoside compounds].

The prolonged use of anti-viral nucleosides (ZDV, ddI, ddC) in HIV-infected patients has given rise to the isolation of viral variants that display resistance against these compounds. Tissue culture selection experiments, involving increasing concentrations of anti-viral compounds, have likewise been shown to select for drug-resistant strains of HIV. Cloning, sequencing and site-directed mutagenesis have shown that a series of point mutations in the viral reverse transcriptase (RT) are responsible for this phenomenon. A different series of mutations in RT are responsible for resistance against non-nucleoside inhibitors of this enzyme. These mutations are due to the error-prone nature of RT during viral replication. Mutated forms of recombinant RT, that derive from drug-resistant viruses, have reduced affinity for relevant triphosphorylated nucleosides.

Acquired Immunodeficiency Syndrome↗

PubMed1995

Development of HIV-1 resistance to (-)2'-deoxy-3'-thiacytidine in patients with AIDS or advanced AIDS-related complex.

OBJECTIVE: To determine the rate of development of in vitro HIV resistance to (-)2'-deoxy-3'-thiacytidine (3TC) and relate the effect of dose to emergence of resistance. METHODS: HIV-infected men and non-pregnant women, aged > or = 18 years, with a CD4 count < or = 300 x 10(6)/l cells were followed in a Phase I/II study, in which they were evaluated for tolerance to 3TC and effect of this agent with regard to viral susceptibility. Peripheral blood and plasma samples were collected at regular intervals for analysis. HIV was isolated using umbilical cord blood mononuclear cells as targets. These cells were also used in determinations of median inhibitory drug concentration. Specific amplification of the 184 mutation site, associated with HIV resistance to 3TC, was performed by polymerase chain reaction, using specific primer pairs, on DNA harvested from infected peripheral blood mononuclear cells (PBMC) of donors or, alternatively, on DNA that had been reverse transcribed from plasma-associated HIV RNA. RESULTS: Phenotypic resistance was detected in approximately one-third of individuals studied, who were followed between 8 and 56 weeks. Development of 3TC resistance occurred independently of dose, although time of first appearance of resistant HIV-1 variants appeared reduced at high 3TC doses. Amino-acid changes at codon 184 in HIV-1 reverse transcriptase were associated with, and preceded, the development of phenotypic 3TC resistance. Most commonly, a Met to Ile substitution appeared transiently before being superceded by a Val substitution at codon 184. CONCLUSIONS: In vitro resistance to 3TC developed in a high proportion of subjects who received prolonged monotherapy with this drug. The development of resistance to 3TC was associated with appearance of mutated viral forms and the disappearance of wild-type virus, with regard to codon 184, in both patient plasma and PBMC.

AIDS-Related Complex↗

PubMed1995

[A study on human behavior and socioeconomic factors affecting malaria transmission and control in Qiongzhong, Hainan].

This study was conducted in Heping District of Quiongzhong County, a hyperedemic mountainous area, in August-September 1992. The comparative surveys between the village and state-run farm, Li and Miao nationalities and Han nationality were carried out by using the sociological method together with the epidemiological methods. Gray relational analysis was conducted between the aforementioned 7 socioeconomic human behavioral factors and IFA rates. The result showed that their degrees of relation (r) were in the following order: (1) percentage of persons who had stayed in the mountain overnight (r = 0.8690); (2) percentage of bed net users (r = 0.7990); (3) percentage of households seeking medical service (r = 0.7990); (4) number of mosquito nets per person (r = 0.7867); (5) percentage of householders knowing malaria transmission route (r = 0.7798); (6) percentage of households with tile-roofed houses (r = 0.6767) and (7) income per capita (r = 0.6636). It indicates that staying in the mountain, using bed net and seeking medical service were three discriminating factors affecting local malaria transmission and control. Therefore, it is suggested that carrying out health education, changing the stay-in-mountain behavior, increasing the utilization of mosquito nets and reinforcing the primary health care should be taken as the fundamental measures for malaria control programme.

China↗

PubMed1995

Structural and ultrastructural studies of the synovial membrane in temporomandibular joint disturbance syndrome.

To gain further knowledge on temporomandibular joint disturbance syndrome (TMJDS), the synovial membranes of 7 patients and 3 cadavers were examined with light microscope and transmission electron microscope. The results showed that the synoviocytes degenerated or disappeared in TMJDS. The cytoplasm contained numerous fibrils, while the organelles diminished in number. The surface of the synovial membrane was covered with a fibrinoid material. The surface of the synovial membrane was covered with a fibrinoid material. The authors consider that these pathological changes reduce or eliminate the normal functions of the synovial membrane, which may lead to damage to the articular cartilage. The articular adhesion found sometimes in TMJDS may be related to the changes in the synovial membrane.

Adult↗

PubMed1995

[The effect of estrogen and its receptor antagonist on the growth of human gastric cancer cell line in vitro].

In this study, the effect was studied of estradiol (E2) and tamoxifen on the growth of a human gastric adenocarcinoma SGC-7901 cell line in vitro. Estrogen receptor as revealed by immunohistochemical staining was positive when added at various concentrations. E2 could significantly stimulate cell growth. The effect was most marked at concentration of 0.1 mumol/L. The cell doubling time was reduced, the plating efficiency was increased and c-myc oncogene expression was enhanced. Flow cytometric analysis indicated that cell] proliferation index was greater in the E2-treated than in the control. All these effects caused by E2 could be partially inhibited by the estrogen receptor inhibitor tamoxifen.

Adenocarcinoma↗

PubMed1995

[Current malaria stratification in China].

During the past 30 years since 1958 when malaria stratification was first made, a great change in malaria situation has occurred. The original stratification can not represent the current status of malaria distribution. Therefore, it needs to be redivided so as to provide basis for future malaria control work. Taking species of mosquito vector and malaria incidence as the main indices and making reference to the natural geographical division, the current malaria endemic area in China can be divided into four regions, i.e. (1) Western region: it is mostly a natural non-endemic area, except some limited areas with sporadic cases in Xinjiang Autonomous Region; (2) Northern region: the whole north-eastern and partial northern China belong to the areas where malaria has been basically eliminated; but sporadic occurrence still exist in most part of the northern China; (3) Central region: it is mostly a hypo-endemic area, only a small part has meso-endemicity; (4) Southern region: nearly 50% has meso-endemicity and the other 50% has hypo-endemicity. The demarcation line of natural non-endemic areas and the areas where malaria has been basically eliminated has been defined.

Animals↗

PubMed1995

[A study on behavioural characteristics of staying on the mountain and its relationship with malaria infection in Li and Miao minorities in Hainan Province].

A study of sociology combined with epidemiology was conducted in Li Minority's two villages and Miao Minority's two villages nearby the foot of mountain in a historically malaria hyperendemic area, Nanqiao Township of Wanning County in August, 1993. The results showed that malaria infection was closely correlated to the behaviour of staying overnight on the mountain of Li and Miao Minorities during farming seasons. Most of Li people stayed overnight on the mountain for planting areca were adults, accounting for 21.6% of the whole population, the duration of stay was about half a year; Miao people stayed overnight on the mountain planting and harvesting upland rice, always with their family member, so the percentage of people who had stayed on the mountain was as high as 82.1%, the duration of stay was about one and half a month. Blood smear examination showed that the malaria parasite rates in both Li and Miao villages were 11.1% and 24.1%, respectively, the positive rate of P. falciparum was 1.0% and 9.1%, respectively, the positive rate of IFA (titer >or= 1 : 40) was 31.2% and 46.5%, respectively. It is indicated that both the rate of parasitaemia and the P. falciparum infection were much higher in Miao Minority than those in Li Minority (P < 0.001). A cohort analysis showed that the malaria parasite rate of population who having not stayed, having ever stayed and being stayed on the mountain was 6.5%, 27.4% and 42.1%, respectively, of them the rate of P. falciparum infection was 0.5%, 8.8% and 18.4%, respectively. The results suggest that malaria acquired from mountainous forest referring to "up-mountain infection" is a major source of malaria infection and a significant risk factor in determining the prevalence of malaria (especially falciparum malaria) in Hainan Province. This behavioural risk factor has become the main obstacle for malaria control in Hainan Province. Therefore, controlling "up-mountain infection" and adopting appropriate anti-malaria measures to protect the risk population who have to work and stay on the mountain should be strengthened in the malaria control program.

China↗

PubMed1995

[Vibratory acoustic stimulation test in fetal hearing monitor].

In order to seek an antepartum diagnosis of hearing impairment in fetus, a vibratory acoustic stimulation test (VAST) was carried out with 57 women who were pregnant 27 weeks and an ABR test was carried out with their 57 newborn babies. The results showed: (1) On or after the 27th week the fetus would have the function of hearing. (2) The reliability of VAST was high (two positive rates were 87.7% and 89.5%) and the VAST was safe. (3) There was a correlation between fetal VAST and newborn ABR (P < 0.05). We consider that VAST is a simple, safe and effective measure for fetal hearing screening.

Acoustic Stimulation↗

PubMed1994

The K65R mutant reverse transcriptase of HIV-1 cross-resistant to 2', 3'-dideoxycytidine, 2',3'-dideoxy-3'-thiacytidine, and 2',3'-dideoxyinosine shows reduced sensitivity to specific dideoxynucleoside triphosphate inhibitors in vitro.

The K65R mutation in human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) encodes cross-resistance to 2',3'-dideoxycytidine (ddC), 2',3'-dideoxy-3'-thiacytidine (3TC), and 2',3'-dideoxyinosine (ddI). We characterized the in vitro sensitivities of recombinant wild type (wt) and K65R mutant RT to dideoxynucleoside triphosphate (ddNTP) inhibitors, using a variety of primer-templates. With poly(rA)-oligo(dT), the K65R mutant showed slight increases in Ki for ddTTP and 3'-azido, 3'-deoxythymidine triphosphate (AZTTP) compared to wt RT, but neither wt nor K65R RT was inhibited by ddCTP or ddATP. With poly(rI)-oligo(dC), the K65R mutant showed a 2-fold increase in Km for dCTP and a 20-fold increase in Ki for ddCTP compared to wt, whereas ddATP, ddTTP, and AZTTP failed to inhibit either enzyme. With a heteropolymeric primer-template, the K65R mutant showed 10-fold reduced sensitivities to ddCTP, 3TCTP, and ddATP, and 4-fold reduced sensitivity to AZTTP, compared to wt. In contrast, both enzymes were equally inhibited by ddTTP and ddGTP. HIV-1 cross-resistance to ddC/3TC/ddI resulting from the K65R mutation may therefore involve selective alterations in substrate/inhibitor recognition. Additionally, competitive inhibition by ddNTPs noncomplementary to the template base appears to be unimportant in the mechanism of inhibition of HIV-1 RT by dideoxynucleoside analogs.

Amino Acid Sequence↗

PubMed1994

Anti-peptidyl transferase leader peptides of attenuation-regulated chloramphenicol-resistance genes.

The chloramphenicol (Cm)-inducible cmlA gene of Tn1696 specifies nonenzymatic resistance to Cm and is regulated by attenuation. The first eight codons of the leader specify a peptide that inhibits peptidyl transferase in vitro. Functionally similar, but less inhibitory, peptides are encoded by the leaders of Cm-inducible cat genes. However, the cat and cmlA coding sequences are unrelated and specify proteins of unrelated function. The inhibition of peptidyl transferase by the leader peptides is additive with that of Cm. Erythromycin competes with the inhibitory action of the peptides, and erythromycin and the peptides footprint to overlapping sites at the peptidyl transferase center of 23S rRNA. It is proposed that translation of the cmlA and cat leaders transiently pauses upon synthesis of the inhibitor peptides. The predicted site of pausing is identical to the leader site where long-term occupancy by a ribosome (ribosome stalling) will activate downstream gene expression. We therefore propose the inducer, Cm, converts a peptide-paused ribosome to the stalled state. We discuss the idea that cooperativity between leader peptide and inducer is necessary for ribosome stalling and may link the activation of a specific drug-resistance gene with a particular antibiotic.

Amino Acid Sequence↗

PubMed1994

Comparison of deoxyoligonucleotide and tRNA(Lys-3) as primers in an endogenous human immunodeficiency virus-1 in vitro reverse transcription/template-switching reaction.

We developed an endogenous in vitro reverse transcription assay to study the properties of priming and template switching during human immunodeficiency virus (HIV) replication. Reactions were primed with HIV reverse transcriptase (RT) and either a deoxyoligonucleotide primer (dPR) or tRNA(Lys-3), the natural primer for reverse transcription. The RNA templates utilized were the actual HIV sequences involved in the first template switch, namely a primer binding sequence (PBS)/U5/R RNA donor template and a R/U3 RNA acceptor template. Reverse transcription reactions using the latter templates and dPR or tRNA(Lys-3) as primers yielded four major products: (-)-strong-stop DNA, a partial template-switched DNA, full template-switched DNA, and a pseudo-PBS-primed product. Use of dPR resulted in three times less template switching than was obtained with tRNA(Lys-3). When reactions were primed with either dPR or tRNA(Lys-3), increases in acceptor:donor template ratios resulted in augmented template switching. Increasing the concentration of RT resulted in increased priming from the PBS but had no effect on the efficiency of template switching. Decreasing the extent of R region overlap resulted in a drop in efficiency of template switching. Decreases in the R region on the donor template also caused a drop in initiation of transcription that was primed by tRNA(Lys-3) from the PBS. In contrast, a corresponding reduction of the R region on the acceptor template had no effect on priming. We conclude that a transcriptional complex of tRNA(Lys-3) and RT may be associated not only with the PBS but also with other cis RNA sequences and secondary structures in a manner essential for efficient priming and template switching.

DNA Primers↗

PubMed1994

Analysis of the secondary structure that negatively regulates inducible cat translation by use of chemical probing and mutagenesis.

Induction of chloramphenicol (Cm) acetyltransferase-encoding genes (cat) by Cm has been proposed to result from the destabilization of a stem-loop that sequesters the ribosome-binding site for the cat coding sequence. Destabilization is caused by the stalling of a ribosome at a specific site in the leader of cat transcripts that immediately precedes the stem-loop. By use of in vivo dimethylsulfate probing of cat-86 leader mRNA, we demonstrate the existence of the stem-loop structure in cat transcripts isolated from uninduced cells and its release during induction. Leader mutations chosen to provide the mRNA with an alternative folding pattern that destabilizes the stem-loop cause constitutive cat expression. Our results establish the occurrence in vivo of the stem-loop in cat-86 transcripts and its role as a negative regulator of cat expression.

Base Sequence↗