Biomedical subjects

T Zhou

Publications and source records attributed to T Zhou.

At least 109 records · Page 6Linked to original sources

PubMed1998

Autocrine and paracrine apoptosis are mediated by differential regulation of Fas ligand activity in two distinct Jurkat T cell populations.

Fas ligand (FasL) produced by activated T cells mediates autocrine-induced apoptosis to limit T cell expansion. To investigate the regulation of FasL activity, Jurkat cells were stably transfected with a 2.3-kb fragment of human FasL promoter that controlled the expression of a GFP reporter gene. Two populations of Jurkat cells with different levels of GFP expression were obtained. One population constitutively expressed high levels of GFP (GFP+), while the other population expressed low levels of GFP (GFP-). The level of GFP expression in the two populations correlated with their levels of FasL transcription and its functional activity. Autocrine regulation of apoptosis was demonstrated by increased FasL activity after stimulation of GFP- cells with anti-CD3, phorbyl myristyl acetate plus ionomycin, or Con A. Paracrine regulation of apoptosis was suggested by the induction of apoptosis of GFP- cells after coculture with unstimulated GFP+ cells. GFP+ cells exhibited a decreased sensitivity to FasL-mediated apoptosis compared with GFP- cells. Furthermore, the cell surface expression of Fas and CD4 was lower on GFP+ cells than GFP- cells, whereas the expression of CD45RO was higher. A decreased level of IL-2 was produced by GFP+ cells after phorbyl myristyl acetate and ionomycin stimulation. Our results indicate that a subpopulation of T cells that express low levels of FasL and IL-2, which are responsive to up-regulation of these molecules after activation, can undergo apoptosis either by suicide after activation or by a paracrine pathway mediated by T cells that constitutively express higher levels of FasL.

Apoptosis↗

PubMed1998

Cell death mediated by Fas-FasL interaction between glial cells and MBP-reactive T cells.

Apoptosis in T cells that have penetrated into the central nervous system (CNS) may be important for the physiological control of T cells with potentially dangerous reactivities to CNS antigens; such control may be dysfunctional in animals suffering from experimental autoimmune encephalomyelitis (EAE). In this study we examined the expression of Fas and FasL genes both in myelin basic protein (MBP)-reactive T cells and in glial cells and the susceptibility of these cells to death induced by Fas/FasL interaction. Both Fas and FasL gene expression is detectable in glial cells and MBP-reactive T cells. Cell death is not unidirectional: when T cells interact with glial cells death can be induced in the former or in the latter population. The ability to induce death of Fas-expressing cells varies greatly among different lines of MBP-reactive T cells, as does resistance to death induction by cells expressing FasL. Moreover, the ability of T cells both to deliver and to resist death signals is a function of their activation status: T cells freshly activated transmit a stronger apoptotic signal to Fas-positive target cells and are also more resistant to FasL-induced suicide. Soluble form of FasL provides a convenient titratable means of delivering death signals via Fas. However, comparison of the susceptibility of different targets to soluble FasL and to FasL expressed on the surface of a transfected glial line revealed differences, suggesting that signals arising from Fas/FasL interaction may be modulated by additional cell-surface molecules.

Animals↗

PubMed1998

Fas/Fas ligand signaling during gestational T cell development.

Most thymocytes express high levels of Fas Ag (Apo-1/CD95); however, the role of Fas/Fas ligand-mediated apoptosis in thymocyte development remains unclear. During gestational development of thymocytes in C57BL/6(B6) +/+ mice, the highest levels of Fas ligand mRNA and Fas ligand protein expression were detected at gestational day (GD) 15, and there was a ninefold decrease in Fas ligand mRNA expression between GD 15 and 17 accompanied by a sixfold increase in Fas mRNA. Apoptotic thymocytes were first detected in the medulla at GD 15, and increasing numbers of cortical clusters and scattered, single apoptotic cells were present on GD 16 and 17. Thus, early apoptosis correlated with high expression of Fas ligand. High levels of Fas ligand mRNA were maintained throughout gestational development in thymocytes of Fas-deficient B6-lpr/lpr mice, but cortical clusters and scattered apoptotic cells were decreased relative to B6 +/+ mice before GD 17. Kinetic analysis of fetal thymic organ cultures treated with anti-Fas Ab demonstrated that thymocytes become sensitive to Fas-mediated apoptosis during the transition from the CD4-CD8- to the CD4+CD8+ phenotype. More mature CD4+CD8+ thymocytes and CD4+ and CD8+ thymocytes became resistant to Fas-mediated apoptosis after GD 17, despite high expression of Fas. However, low avidity engagement of the TCR on Fas-sensitive CD4+CD8+ thymocytes before GD 17 induced resistance to Fas-mediated apoptosis. The present results indicate that Fas plays a critical role in mediating apoptosis during early gestational thymocyte development and that thymocytes that receive a survival signal through TCR/CD3 become resistant to Fas-mediated apoptosis.

Animals↗

PubMed1998

Reduction of arthritis and pneumonitis in motheaten mice by soluble tumor necrosis factor receptor.

OBJECTIVE: To determine the effects of anti-tumor necrosis factor (anti-TNF) therapy in the inflammatory and autoimmune disease in motheaten (me/me) mice, which exhibit a Fas apoptosis signaling defect. METHODS: Arthritis, pneumonitis, and mortality were analyzed in me/me mice treated with a novel, soluble, dimeric TNF receptor I (sTNFRI) molecule capable of high-affinity binding and neutralization of TNFalpha. RESULTS: Soluble TNFRI reduced serum levels of TNFalpha and led to a 2-fold increase in the lifespan of me/me mice, compared with the control treatment group. The treatment also reduced the development of the "motheaten" skin patches and alleviated pneumonitis and inflammatory lesions in the extremities of me/me mice compared with controls. However, the serum levels of IgM and IgM anti-double-stranded DNA autoantibody were comparable to those of untreated control mice. CONCLUSION: TNFalpha is an important cytokine involved in the pathogenesis of inflammatory disease in me/me mice, resulting in tissue damage and early mortality. Therapies directed at blocking TNF/TNFR interactions, such as the sTNFRI used in these experiments, may be effective in diseases associated with apoptosis defects leading to overutilization of the TNF/TNFR pathway.

Alopecia↗

PubMed1998

Murine cytomegalovirus induces a Sjögren's syndrome-like disease in C57Bl/6-lpr/lpr mice.

OBJECTIVE: To analyze Fas and tumor necrosis factor receptor I (TNFRI) apoptosis pathways in salivary gland inflammatory disease induced by murine cytomegalovirus (MCMV) infection. METHODS: Four different strains of mice (C57BI/6 [B6]-+/+, Fas-deficient B6-lpr/lpr, TNFRI-deficient B6-tnfr1(0/0), and B6-tnfr1(0/0)-lpr/lpr mice) were infected intraperitoneally with the Smith strain of MCMV (1 x 10(5) plaque-forming units). Viral load was determined by a plaque assay, inflammation and apoptosis by immunohistochemistry and staining with terminal dUTP nickend labeling, and autoantibodies by enzyme-linked immunosorbent assay. RESULTS: Infectious MCMV was not detectable by day 100. Although all MCMV-infected mice developed acute sialadenitis by day 28, a chronic (>100 days), severe salivary gland inflammation and anti-Ro and anti-La antibodies developed only in the B6-lpr/lpr mice. Apoptotic cells were detected during the acute, but not the chronic, phase of inflammation. CONCLUSION: Both Fas- and TNFRI-mediated apoptosis contribute to the clearance of MCMV-infected cells in the salivary glands. However, because Fas-mediated apoptosis is necessary for the down-modulation of the immune response, a defect in this process can lead to a postinfection, chronic inflammatory response that resembles Sjögren's syndrome.

Animals↗

PubMed1998

Production of rat salivary cystatin S variant polypeptides in Escherichia coli.

Cystatins are protein inhibitors of papain and related cysteine proteinases. A series of continuous synthetic peptides corresponding to the entire sequence of rat salivary cystatin was used to localize the binding domains of the cystatin to papain. Several synthetic peptides, one from the aminoterminal sequence (peptide 1-24) and others from the carboxylterminal (peptides 66-79, 66-90, 79-90, 79-114), showed binding to papain, but none of the peptides showed inhibition of papain activity. Three recombinant rat salivary cystatin variants (N-terminal truncated protein lacking amino acid residues 1-9; variant 49-53, in which amino acid residues QVVAG of rat salivary cystatin had been replaced with amino acid residues LVL in mutant protein; and variant 65-78, in which amino acid residues 65-78 had been replaced with amino acids PG in mutant protein) were produced using the Escherichia coli expression system pGex-4T. To generate N-terminal truncated protein the desired coding region of the cystatin gene was amplified by polymerase chain reaction (PCR). To produce the variants 49-53 and 65-78, a PCR-based approach of gene splicing by overlap extension was used. Recombinant cystatin proteins were produced as insoluble inclusion bodies as fusion proteins with a glutathione S-transferase (GST) carrier. After solubilization with urea the GST carrier was cleaved from the fusion protein with thrombin and cystatin variants purified by fast liquid chromatography on a MonoQ column. The purified proteins reacted with antibodies to rat salivary cystatin. The N-terminal truncated and variant 49-53 exhibited very little inhibitory activity towards papain, whereas variant 65-78 exhibited papain-inhibitory activity similar to the full-length recombinant cystatin.

Alternative Splicing↗

PubMed1998

Calmodulin antagonists inhibit apoptosis of CD4+ T-cells from patients with AIDS.

Recent studies indicate that Fas and Fas ligand are involved in apoptosis of T-cells in HIV-infected patients. We have demonstrated that calcium/calmodulin is involved in Fas-mediated apoptosis in human T-cell lines transfected with HIV recombinant cDNA. In the present study, we examined spontaneous apoptosis of T-cells in vitro in peripheral blood obtained from 11 patients with AIDS and 8 HIV-seronegative normal donors and the effect of the calmodulin antagonists, trifluoperazine (TFP) or tamoxifen (TMX), on apoptosis. The results show that: (1) levels of spontaneous apoptosis were higher in PBMCs obtained from patients with AIDS than HIV-negative normal controls and the levels of apoptosis correlated with the severity of disease. (2) The accelerated apoptosis occurred predominantly in CD4+ cells in patients with AIDS. (3) Calmodulin antagonists inhibited the spontaneous apoptosis of CD4+ T-cells from patients with AIDS, which resulted in an increase in the ratio of CD4+ to CD8+ T-cells. (4) The inhibitory effect of calmodulin antagonists on apoptosis was more significant in patients with advanced disease (CDC category C) compared to less severe disease (CDC category B). These results indicate that calmodulin antagonists inhibit HIV-associated apoptosis of CD4+ T-cells, and imply that the calcium/calmodulin play important roles in mediating apoptosis of CD4+ T-cells induced by HIV infection.

Acquired Immunodeficiency Syndrome↗

PubMed1998

Induction of specific T-cell tolerance by adenovirus-transfected, Fas ligand-producing antigen presenting cells.

A major problem associated with adenovirus gene therapy is the T cell-mediated immune response, which is elicited by inoculation of the adenovirus vector and leads to rapid clearance of the virus and loss of transgene expression. In this study, the immune response to adenovirus was prevented by induction of specific T-cell tolerance by pretreatment with adenovirus-infected antigen-presenting cells (APC) that express Fas ligand. Compared with control-treated mice, the tolerized mice showed prolonged expression of lacZ upon administration of AdCMVlacZ 1 week after tolerance induction. In contrast to the control mice, the tolerized mice did not display proliferation of CD3+ T cells in the spleen in response to AdCMVlacZ. Tolerance induction also was indicated by the lower production of interferon-gamma and interleukin-2 by peripheral T cells isolated from AdCMVlacZ-challenged tolerized mice than by AdCMVlacZ-challenged control-treated mice. The T-cell tolerance was specific for the adenovirus as the T-cell responses to irrelative murine cytomegalovirus remained unimpaired. Our results indicate that adenovirus-specific T-cell tolerance can be induced by APCs that coexpress Fas ligand and adenovirus antigens. We propose that this new strategy can be used to induce tolerance to adenovirus vector gene therapy with resultant prolonged expression of the transgene.

Adenoviridae↗

PubMed1998

Metalloid resistance mechanisms in prokaryotes.

Resistance to antibiotics and other chemotherapeutic agents is becoming a wide spread health issue. The biochemical mechanisms of resistance vary, but active efflux of the toxic agents is one of the most common. Bacterial resistances to metals provide good model systems for transport-related resistances. One of the best understood metal resistance systems is the product of the ars operon, which provides resistance to arsenicals and antimonials. As a reflection of the ubiquity of arsenic in the environment, ars operons are found in all species of bacteria, carried in chromosomes, plasmids, and transposons. This review focuses on the biochemistry of the proteins of the ars operon of R-factor R773. The system is novel in several respects. First, it is regulated at the transcriptional and allosteric levels, and regulation is effected through cysteine thiol interaction with As(III) or Sb(III). Thus soft metal-thiol chemistry provides a high affinity digital switch to turn the regulated protein on with rapidity. The transport system that provides resistance, on the other hand, uses oxyanions of arsenic or antimony as substrates. This nonmetal chemistry allows for low affinity interactions of the membrane transporter with substrate, conductive with translocation and release of substrate on the outside of the cell membrane. Second, the transporter is uniquely capable of coupling to either electrochemical energy as a secondary carrier protein or the chemical energy of ATP when binding of a catalytic subunit converts it into an anion-translocating ATPase.

Adenosine Triphosphatases↗

PubMed1998

Application of a Fas ligand encoding a recombinant adenovirus vector for prolongation of transgene expression.

An adenovirus vector encoding murine Fas ligand (mFasL) under an inducible control was derived. In vivo ectopic expression of mFasL in murine livers induced an inflammatory cellular infiltration. Furthermore, ectopic expression of mFasL by myocytes did not allow prolonged vector-mediated transgene expression. Thus, ectopic expression of functional mFasL in vector-transduced cells does not appear to confer, by itself, an immunoprivileged site sufficient to mitigate adenovirus vector immunogenicity.

Adenoviridae↗

PubMed1998

Detection of the low density lipoprotein receptor gene PvuII intron 15 polymorphism using the polymerase chain reaction: association with plasma lipid traits in healthy men and women.

We have used anchored PCR to amplify and sequence 1400 bp of the 15th intron of the Low Density Lipoprotein (LDL) receptor gene, and have determined oligonucleotides and conditions for the genotyping of the previously reported PvuII polymorphism. The cutting site (CAGCTG) is created by the transition of a CpG to a TpG within the sequence CAGCCG at a position roughly 600 bp 5' from the splice acceptor site of exon 16. Genotype was determined in three population-based samples of healthy individuals. In a group of 318 men and women from Iceland the frequencies of the Intron-15 T (cutting) allele was 0.23 (95% CI, 0.19-0.28) and was similar in men and women. In two groups of men from England (n = 385) and Scotland (n = 320), the frequency was similar, being 0.23 (0.19-0.27) and 0.25 (0.22-0.28) respectively. Individuals who were homozygous for the T allele had lower levels of total-cholesterol triglycerides and apolipoprotein B, than those with other genotypes, and in the combined group of UK men this effect reached statistical significance; compared to the C/C group, the T/T group had 6% lower cholesterol (p = 0.02) and 15% lower triglycerides (p = 0.03). The lowering effect associated with the T/T genotype was greater in men who were in the lowest tertile of body mass index (< 25 kg/m2) and for the trait of apoB levels, this genotype x obesity interaction was statistically significant (p = 0.01). We thus confirm the association between this allele and lower levels of plasma lipid levels previously reported. The availability of a PCR-based method to detect this polymorphism will facilitate further investigation of the impact of LDL-receptor gene variation in determining lipid levels.

Alleles↗

PubMed1998

[Investigation on sera prevalence rate of varicella and immunogenicity of varicella vaccine in healthy children].

A whole varicella virus antigen-ELISA method was used to investigate the sera prevalence rate of varicella in 315 healthy children aged 3-7 years. Results showed that the sera positive rate of anti-VZV-IgG was 41.6%. Different prevalence rates in different kindergartens were noticed, characterizing the differences of populations. Immunogenicity of Oka strain attenuated varicella vaccine among children was also studied. No obvious adverse reactions were observed and the seraconversion rate was 85.1%.

Antibodies, Viral↗

PubMed1998

[Comparison of the effects of inhaled nitric oxide and intravasculare regitine on pulmonary gas exchange in young dogs with oleic-acid acute lung injury].

In order to find an agent which truly improves hypoxemia of some serious pediatric lung diseases, the authors examined the independent effects of nitric oxide inhalation and regitine infusion on blood gases, intrapulmonary shunt and hemodynamics in young dogs with oleic-acid acute lung injury. After nitric oxide inhalation, the results showed moderate increases in PaO2 and SaO2 (P > 0.05) and a significant decrease in Qs/Q tau ratio (P < 0.01). There was a significant decrease of PAP(P < 0.05), while SAP remained unchanged. After regitine infusion, however, there were marked decreases in PaO2 and SaO2 (P < 0.01); meanwhile, Qs/Q tau rose (P < 0.05). These suggest that with the presence of pulmonary pathology nitric oxide inhalation may alleviate the elevated pulmonary pressure without alteration in systemic artery pressure; so it can improve pulmonary ventilation-perfusion distribution and cause favorable changes in blood gases. On the other hand, regitine, as a non-selective vasodilator, reduces pulmonary artery pressure at the cost of significant worsening of blood oxygenation and systemic hypotension; so its routine use in childhood pulmonary diseases should be cautiously considered.

Animals↗

PubMed1998

[Voice measurement with computer and analysis in normal adult].

Acoustic parameters of 90 normal adults (20-50 years old) were measured with computer. The results showed that fundamental frequency (F0), frequency perturbation quotient (FPQ) and amplitude perturbation quotient (APQ) between male and female in same ages were statistically significant difference. F0 and FPQ were bigger but APQ was smaller in female. FPQ and APQ were significantly related to F0. OCT wasn't obviously different among sexes or ages. The article indicated that voice of adults and the vibration of vocal cords had distinctive characteristics in different sexes or mode of phonation, which could provide objective methods and grounds for clinical voice evaluation.

Adult↗

PubMed1998

[Experimental study on human collagen and autologous fat injected into canine vocal cords].

OBJECTIVE: To explore the histological fate of human collagen and autologous fat injected into vocal cords in treating glottic insufficiency. METHODS: Human collagen and autologous fat were injected respectively into two dogs(group A) to correct glottic incompetence caused by left vocal cord paralysis. Autologous fat and human collagen 0.3 ml were injected respectively into left and right vocal cords of 9 dogs (B, C, D groups, 3 dogs in each group). These animals were killed after 1, 3 and 5 months. The histology, volume and curative effect of both substances were evaluated. RESULTS: Autologous fat grafts survived in the canine vocal cords after 5 months but the absorption was rapid. The average speed of reduction in volume was 38.51% from 1 to 5 months and the effect of treatment couldn't last long. Human collagen was easily injected and well-tolerated. The injected collagen was colonized by host fibroblasts and capillaries. At the same time, new host collagen deposited. There was a trend to allow the eventual replacement of the implant by new host tissue. The average speed of reduction in volume was 0.65% and the curative effect was constantly stable from 1 to 5 months. CONCLUSION: The results suggested that autologous fat has only a potential use for short-term(< 5 months) augmentation of vocal cord. Human collagen can be used as long-term augmentative material to manage glottic insufficiency caused by various etiologies.

Adipose Tissue↗

PubMed1998

[Studies on urinary metabolites of HH07A, a derivative of hainanensine, in rats].

The metabolism of HH07A in the rat has been investigated by GC-MS. After oral administration of HH07A, the rat urine was passed through a macroporous XAD-2 resin column and eluted with methanol. The methanol extract was hydrolyzed with glucuronidase, extracted with dichloromethane and concentrated for TMS derivatization with GC-MS. The mass spectra of HH07A and its four metabolites as well as their derivatives were presented. The structures of the metabolites were proposed and the in vivo metabolic pathway of HH07A was given in Figure 6.

Animals↗

PubMed1998

[Studies on the fine stereostructure of taxoids].

The anticancer drug taxol is a new type antimicrotubular drug. The structure-activity relationship study on taxol and its analogs has indicated the importance of the C13 side chain and the C4-C5-C20 epoxypropane group. The crystal structures of 4 taxoids having the basic skeleton with 5/7/6 membered ring and three types of 10 taxoids having the basic skeleton with 6/8/6 membered ring have been analyzed. The effects of the induction by different substructures at C4 on the stereostructures of taxoids were studied, and the relationship between the 6/8/6/4 skeleton and the anticancer activity was discussed from the crystallographic point.

Antineoplastic Agents, Phytogenic↗

PubMed1998

[Studies on analytical method for 10 drugs of abuse in urine using HPLC].

A systematic determination method for 10 drugs of abuse, morphine, codeine, 6-monoacetylmorphine, heroin, levorphanol, pethidine, ethylmorphine, anadol, pentazocine and ethamivan, using high performance liquid chromatography is described. Separation of the 10 drugs was achieved on a 25 cm x 4.6 mm ID, 10 microns Zorbax C8 column, using methanol--0.05 mol.L-1 KH2PO4--diethyl amine (27:73:0.5, pH 4) as mobile phase. Codeine was used as the interal standard. Calibration graphs were linear over the concentration range of 10-200 micrograms.ml-1 and regression equations showed correlation coefficients of greater than 0.999. Precision (RSD) were found to be better than 3% for each compound. Precision and linearity of the method are satisfactory for clinical toxicological applications. The extraction procedure yielded cleaner extracts. The recovery rates from urine were all above 87% without interference. This method is rapid, sensitive and specific.

Chromatography, High Pressure Liquid↗