Biomedical subjects

X Zhou

Publications and source records attributed to X Zhou.

At least 487 records · Page 27Linked to original sources

PubMed1998

[Determination of zinc, copper, iron and manganese contents in hair for MPA patients and healthy men].

In this paper the contents of zinc, copper, iron and manganese in hair for MPA patients and 38 hair-healthy men have been determined by FAAS. The results show that the hair zinc and manganese levels in MPA patients are significantly lower than that in hair-healthy men. The hair copper level in MPA patients is significantly higher than that in hair-healthy men. There are statistic difference (P < 0.05) in hair zinc, copper and menganese levels among two groups. There is no statistic difference (P > 0.05) in the iron level among the two gopups. This paper provides useful material for the research of the cause and treatment of the MPA.

Alopecia↗

PubMed1998

[FTIR study on the normal and tumor gastrointestinal tissues].

Series of cancer tissues and corresponding normal tissues of gastrointestinal tract (stomach, colon, esophagus) were studied by FTIR technique and the results showed the analogy of the spectra for the cancer tissues, while the spectra of normal tissues can be classified into three kinds. The secondary structures of protein were obtained by using deconvolution and curve-fitting techniques with Amide I bands and the results showed that the contents of alpha -helix and beta-sheet are different between the normal and cancer tissues.

Connective Tissue↗

PubMed1998

Transmission and natural regulation of infection with Ascaris lumbricoides in a rural community in China.

A longitudinal study on a cohort of 215 people who remained untreated from June 1993 to June 1994 was carried out using fecal egg counts to examine the course of transmission and aspects of the natural regulation of the infection with Ascaris lumbricoides. The results indicate that the parasite population remained relatively stable during the years since overall prevalence remained steady above 60%. About 40% of the people in the cohort retained their infection intensity status throughout the year, in terms of light, medium, and heavy infections. The regulation of infection in the population could be attributed to the tendency for the loss of worms from people with initial light infections and the movement to medium infection intensity status of people with both light and heavy infections. Compared with adults, children showed a stronger tendency of retaining initial medium infections but weaker tendencies for keeping initial light infections and clearing themselves of infection. Predisposition to light and medium infections was detected in people who had experienced an uninfected period of 4-12 mo before reinfection was established.

Adolescent↗

PubMed1998

Expression of apoptosis regulators in cutaneous malignant melanoma.

Metastatic malignant melanoma (MM) is usually incurable and responds poorly to chemotherapy. Because many cytotoxic drugs cause cell death by inducing apoptosis, an imbalance of apoptosis regulatory proteins may contribute to MM treatment resistance. We have previously shown reduced expression of Bcl-2 protein, a negative regulator of apoptosis, in MM as compared with benign nevi. It is hypothesized that other apoptosis regulators may be involved in survival of MM cells. We examined the expression of Bax, Bcl-2, Bcl-X, and Mcl-1 in human benign nevi, primary MM, and metastatic MM using immunohistochemistry. Results were confirmed with Western blotting. The proapoptotic protein, Bax, was surprisingly overexpressed in all MM samples compared with benign nevi. Interestingly, in most MM samples there was overexpression of Mcl-1 or Bcl-XL, both negative regulators of apoptosis. Increased expression of Mcl-1 and Bcl-XL was first observed in thin primary melanomas, suggesting that up-regulation of these proteins represents a relatively early event associated with malignant transformation in MM. As published previously, the majority of primary and metastatic MM exhibited reduced Bcl-2 levels. We conclude that the apoptosis inhibitors Bcl-XL or Mcl-1, alone or in combination, may circumvent the normal cell death pathway, contributing to the pathogenesis and treatment resistance in metastatic MM.

Apoptosis↗

PubMed1998

Colonial growth and differentiation of epithelial cells derived from abattoir adult porcine livers.

The parenchymal cell fraction was isolated from abattoir adult porcine livers and cultured in Dulbecco's Modified Eagles' medium/Ham's F12 medium (DMEM/F12; 1:1) medium supplemented with 5% foetal calf serum, 10 ng/mL glucagon, 10 microg/mL insulin, 60 ng/mL hydrocortisone and eight other factors (NAIR-1 medium). The fraction contained a number of epithelial cells other than hepatocytes, some of which attached to the culture plates as cell clusters and began to grow after 3 days in culture. These epithelial cells growing as colonies were found to express cytokeratin 18 by immunocytochemistry. After 7-8 days, duct-like structures emerged in the central parts of the colonies. The cells constituting the duct-like structures and some cells located outside the structures were positive for cytokeratin 19 and gamma-glutamyltransferase (GGT). The albumin-positive cells were located in the outer parts of the colonies rather than their central parts. Albumin was also detectable in the cells surrounded by the duct-like structures. Moreover, cytochrome P450 IA1 was induced by 3-methylcholanthrene (3-MC) on day 16. These results suggest that porcine liver epithelial cell clusters may contain stem-like cells which can differentiate into mature hepatocytes or bile duct epithelial cells.

Albumins↗

PubMed1998

Mechanism of suppression of natural killer cell activity in trauma patients.

Trauma patients develop a severe immunosuppression that includes suppression of natural killer (NK) cell activity although numbers of NK cells are not reduced. The mechanism of suppression of NK cell activity after major trauma is not known. The aim of the present study was to investigate the in vitro effect of plasma samples from trauma patients (TP) on the cytotoxic activity of normal NK cells. Buffycoat mononuclear cells (5x10(5)/well) were preincubated with either TP or plasma samples from age and sex matched healthy controls (CP) for 0, 16 or 40 h. These effector cells were then cultured with 51Cr labeled K-562 cells (2x10(4)/well) for 4 h at 37 degrees C and % lysis was calculated. No significant differences in % lysis between CP and TP were found with 0 or 16 h preincubation, however 40 h preincubation with TP severely suppressed NK cell function (p=0.003) as compared to preincubation with CP for the same period. Addition of neutralizing anti-IL-4, anti-TGF-beta1, or anti-IL-10 antibodies did not reverse the NK cell suppression. There was a partial reversal of NK cell suppression by catalase but not by SOD or L-NMMA. Removal of monocytes from buffycoat mononuclear cells also significantly reversed the NK cell suppression. These data suggest that suppression of NK cell activity in trauma patients may be an accessory cell dependent phenomenon and may partially depend on production of reactive oxygen metabolites (ROM).

Catalase↗

PubMed1997

Expression and characterization of recombinant murine lactoferrin.

Lactoferrin (Mw=78 kDa) is a member of the transferrin family of iron-binding glycoproteins. Previous studies carried out primarily in vitro indicate that the protein has multifunctional properties and may be involved in regulation of iron homeostasis, inhibition of bacterial growth and regulation of immune responses. However, the significance and species specificity of these proposed functions in vivo have not been adequately addressed due to lack of sufficient purified homospecies lactoferrin for analysis in small animal models. We previously reported the successful production of biologically active recombinant human lactoferrin using an Aspergillus expression system. In the present study, we report the production of recombinant murine lactoferrin using a similar expression strategy. Recombinant murine lactoferrin was purified to homogeneity and was similar in size and immunoreactivity to native murine milk lactoferrin. The recombinant protein was correctly processed at its N-terminus and was glycosylated. Interestingly, while both human and murine lactoferrin bind iron in a 2:1 molar ratio, iron bound to recombinant murine lactoferrin was more acid labile than human lactoferrin, demonstrating species-specific variation in the stability of iron-binding to this protein. Finally, the availability of recombinant murine lactoferrin will now facilitate the study of the species specificity of lactoferrin action in a mouse model system.

Amino Acid Sequence↗

PubMed1997

Heat shock fusion proteins as vehicles for antigen delivery into the major histocompatibility complex class I presentation pathway.

Mice immunized with heat shock proteins (hsps) isolated from mouse tumor cells (donor cells) produce CD8 cytotoxic T lymphocytes (CTL) that recognize donor cell peptides in association with the major histocompatibility complex (MHC) class I proteins of the responding mouse. The CTL are induced apparently because peptides noncovalently associated with the isolated hsp molecules can enter the MHC class I antigen processing pathway of professional antigen-presenting cells. Using a recombinant heat shock fusion protein with a large fragment of ovalbumin covalently linked to mycobacterial hsp70, we show here that when the soluble fusion protein was injected without adjuvant into H-2b mice, CTL were produced that recognized an ovalbumin-derived peptide, SIINFEKL, in association with Kb. The peptide is known to arise from natural processing of ovalbumin in H-2b mouse cells, and CTL from the ovalbumin-hsp70-immunized mice and a highly effective CTL clone (4G3) raised against ovalbumin-expressing EL4 tumor cells (EG7-OVA) were equally effective in terms of the concentration of SIINFEKL required for half-maximal lysis in a CTL assay. The mice were also protected against lethal challenge with ovalbumin-expressing melanoma tumor cells. Because large protein fragments or whole proteins serving as fusion partners can be cleaved into short peptides in the MHC class I processing pathway, hsp fusion proteins of the type described here are promising candidates for vaccines aimed at eliciting CD8 CTL in populations of MHC-disparate individuals.

Amino Acid Sequence↗

PubMed1997

Impaired mast cell-dependent natural immunity in complement C3-deficient mice.

The complement system is widely regarded as essential for normal inflammation, not least because of its ability to activate mast cells. However, recent studies have called into question the importance of complement in several examples of mast cell-dependent inflammatory responses. To investigate the role of complement in mast cell-dependent natural immunity, we examined the responses of complement-deficient mice to caecal ligation and puncture, a model of acute septic peritonitis that is dependent on mast cells and tumour necrosis factor-alpha (TNF-alpha). We found that C4- or C3-deficient mice were much more sensitive to caecal ligation and puncture than wild-type (WT) controls (100% versus 20% in 24-h mortality, respectively). C3-deficient mice also exhibited reductions in peritoneal mast cell degranulation, production of TNF-alpha, neutrophil infiltration and clearance of bacteria. Treating the C3-deficient mice with purified C3 protein enhanced activation of peritoneal mast cells, TNF-alpha production, neutrophil recruitment, opsonophagocytosis of bacteria and resistance to caecal ligation and puncture, confirming that the defects were complement-dependent. These results provide formal evidence that complement activation is essential for the full expression of innate immunity in this mast cell-dependent model of bacterial infection.

Animals↗

PubMed1997

Vena contracta imaged by Doppler color flow mapping predicts the severity of eccentric mitral regurgitation better than color jet area: a chronic animal study.

OBJECTIVES: This study sought to evaluate the relation between the color Doppler-imaged vena contracta and the severity of mitral regurgitation (MR) in a chronic animal model of MR. BACKGROUND: The vena contracta, which is defined as the smallest connection between the laminar flow acceleration zone and the turbulent regurgitant jet, has been reported to be a clinically useful marker for evaluating the severity of valvular regurgitation. METHODS: Six sheep with chronic MR produced by previous operation severing the chordae tendineae were examined. MR jet flows and vena contracta widths were imaged using a Vingmed 775 scanner with a 5-MHz transducer. Image data were directly transferred in digital format to a microcomputer for off-line measurement. MR was quantified as peak and mean regurgitant flow rates, regurgitant stroke volumes and regurgitant fractions determined using mitral and aortic electromagnetic flow probes and flowmeters balanced against each other. RESULTS: Vena contracta width correlated well with regurgitant severity determined by electromagnetic flowmeters (r = 0.95, SEE = 0.05 cm, p < 0.0001 for peak regurgitant flow rate; r = 0.85, SEE = 0.08 cm, p < 0.0001 for regurgitant stroke volume; r = 0.90, SEE = 0.07 cm, p < 0.0001 for regurgitant fraction). CONCLUSIONS: This study shows that the vena contracta width method is useful for predicting the severity of MR. It is simple and conveniently available with high resolution equipment. The quantitative comparisons in the present study lay the foundation for future clinical and research studies using this vena contracta technique.

Animals↗

PubMed1997

Nerve growth factor-specific regulation of protein methylation during neuronal differentiation of PC12 cells.

Protein methylation is a posttranslational modification that can potentially regulate signal transduction pathways in a similar manner as protein phosphorylation. The role of protein methylation in NGF signaling was examined by metabolic labeling of PC12 cell proteins with L-[methyl-3H]methionine and by in vitro labeling of cell proteins with L-[methyl-3H]S-adenosylmethionine. Effects of NGF were detected within 15 min. Methyl-labeled proteins were resolved by one and two dimensional SDS-PAGE. NGF affected the methylation of several 68-60-kD proteins (pI 5.8-6.4) and 50-kD proteins (isoelectric point pH 6.7-6.8 and 5.8-6.2). Several NGF-induced changes in methylation increased over several hours and through 4 d. Moreover, methyl labeling of several specific proteins was only detected after NGF treatment, but not in nontreated controls. The effects of NGF on protein methylation were NGF specific since they were not observed with EGF or insulin. A requirement for protein methylation for neurite outgrowth was substantiated with either of two methylation inhibitors: dihydroxycyclopentenyl adenine (DHCA) and homocysteine. DHCA, the more potent of the two, markedly inhibits protein methylation and neurite outgrowth without affecting cell growth, NGF-induced survival, cell flattening, or several protein phosphorylations that are associated with early signaling events. Removal of DHCA leads to rapid protein methylation of several proteins and concurrent neurite outgrowth. The results indicate that NGF regulates the methylation of several specific proteins and that protein methylation is involved in neurite outgrowth from PC12 cells.

Adenosine↗

PubMed1997

A major role for neutrophils in experimental bullous pemphigoid.

Bullous pemphigoid (BP) is an inflammatory subepidermal blistering disease associated with an IgG autoimmune response to the hemidesmosomal protein, BP180. Using a passive transfer mouse model, our group has shown previously that antibodies to the murine BP180 (mBP180) ectodomain are capable of triggering a blistering skin disease that closely mimics human BP. In this study, we investigated the role of neutrophils in the immunopathogenesis of this disease model. BALB/c mice depleted of circulating neutrophils by treatment with neutrophil-specific antibodies were no longer susceptible to the pathogenic effects of anti-mBP180 IgG. IgG and complement were deposited at the dermal-epidermal junction of these animals, but there was no evidence of inflammatory infiltration or blistering. C5-deficient mice, which are resistant to the pathogenic activity of anti-mBP180 IgG, could be made susceptible to this IgG-mediated blistering disease by intradermal administration of a neutrophil chemoattractant, IL-8 or C5a. Intraperitoneal injection of IL-8, which sequesters neutrophils in the peritoneal cavity, interferes with anti-mBP180-induced neutrophilic infiltration of the skin and prevented the development of BP disease in BALB/c mice. These findings provide the first direct evidence that neutrophils recruited to the skin via a C5-dependent pathway play an essential role in subepidermal blister formation in experimental BP, and suggest new directions for disease intervention.

Animals↗

PubMed1997

Expression, purification, and characterization of inosine 5'-monophosphate dehydrogenase from Borrelia burgdorferi.

Inosine 5'-monophosphate dehydrogenase (IMPDH) is the rate-limiting enzyme in de novo guanine nucleotide biosynthesis. IMPDH converts IMP to xanthosine 5'-monophosphate with concomitant conversion of NAD+ to NADH. All IMPDHs characterized to date contain a 130-residue "subdomain" that extends from an N-terminal loop of the alpha/beta barrel domain. The role of this subdomain is unknown. An IMPDH homolog has been cloned from Borrelia burgdorferi, the causative agent of Lyme disease (Margolis, N., Hogan, D., Tilly, K., and Rosa, P. A. (1994) J. Bacteriol. 176, 6427-6432). This homolog has replaced the subdomain with a 50-residue segment of unrelated sequence. We have expressed and characterized the B. burgdorferi IMPDH homolog. This protein has IMPDH activity, which unequivocally demonstrates that the subdomain is not required for catalytic activity. The monovalent cation and dinucleotide binding sites of B. burgdorferi IMPDH are significantly different from those of human IMPDH. Therefore, these sites are targets for the design of specific inhibitors for B. burgdorferi IMPDH. Such inhibitors might be new treatments for Lyme disease.

Binding Sites↗

PubMed1997

Beta2-microglobulin-deficient mice are resistant to bullous pemphigoid.

Recent understanding of the mechanism of immunoglobulin G (IgG) catabolism has yielded new insight into antibody-mediated diseases. We proposed that beta2-microglobulin (beta2m)-deficient mice have been protected from systemic lupus erythematosis (SLE)-like syndromes because they lack the beta2m-associated IgG protection receptor (FcRn) and therefore catabolize IgG, including pathogenic IgG autoantibodies, considerably more rapidly than normal mice. Such an hypothesis would predict that beta2m-deficient mice would also be resistant to experimental bullous pemphigoid, a disease with a pathogenesis thought to be much simpler than SLE, being the result of antibody directed toward a pathogenic epitope on the epidermal hemidesmosome that anchors basal keratinocytes to the basement membrane. To test this hypothesis, we administered pathogenic rabbit antibody directed toward the hemidesmosome to beta2m-deficient mice and to normal control mice, both intraperitoneally and intradermally, and assessed the mice clinically, histologically, and immunologically for manifestations of skin disease. We found that the beta2m-deficient mice were protected when the antibody was given intraperitoneally whereas intradermal administration resulted in blisters only slightly less severe than those seen in normal mice. These data would indicate that autoantibody-mediated inflammation might be prevented or controlled by appropriate modulation of FcRn function.

Animals↗

PubMed1997

Regulation of protein turnover by glutamine in heat-shocked skeletal myotubes.

Skeletal muscle accounts for approximately one-half of the protein pool in the whole body. Regulation of protein turnover in skeletal muscle is critical to protein homeostasis in the whole body. Glutamine has been suggested to exert an anabolic effect on protein turnover in skeletal muscle. In the present work, we characterized the effect of glutamine on the rates of protein synthesis and degradation in cultured rat skeletal myotubes under both normal and heat-stress conditions. We found that glutamine has a stimulatory effect on the rate of protein synthesis in stressed myotubes (21%, P < 0.05) but not in normal-cultured myotubes. Glutamine shows a differential effect on the rate of degradation of short-lived and long-lived proteins. In both normal-cultured and stressed myotubes, the half-life of short-lived proteins was not altered while the half-life of long-lived proteins increased with increasing concentrations of glutamine in a concentration-dependent manner. In normal-cultured myotubes, when glutamine concentration increased from 0 to 15 mM, the half-life of long-lived proteins increased 35% (P < 0.001) while in stressed myotubes, it increased 27% (P < 0.001). We also found that glutamine can significantly (P < 0.001) increase the levels of heat-shock protein 70 (HSP70) in stressed myotubes, indicating that HSP 70 may participate in the mechanism underlying the effect of glutamine on protein turnover. We conclude that in cultured skeletal myotubes the stimulatory effect of glutamine on the rate of protein synthesis is condition-dependent, and that the inhibitory effect of glutamine on the rate of protein degradation occurs only on long-lived proteins.

Animals↗

PubMed1997

1-O-Octadecyl-2-O-methylglycerophosphocholine inhibits protein kinase C-dependent phosphorylation of endogenous proteins in MCF-7 cells.

Studies with leukaemic cells, based primarily on in vitro assays, have suggested that antitumour ether lipids have only a moderate effect on protein kinase C (PKC) activity, and, furthermore, inhibition of PKC is unlikely to be involved in the mechanism of inhibition of cell proliferation by these compounds. To determine if this is also the case for epithelial cancer cells, we examined the effect of 1-O-octadecyl-2-O-methylglycerophosphocholine (ET18-OCH3) on PKC-induced phosphorylation of endogenous proteins in MCF-7 cells under incubation conditions where the drug inhibited cell proliferation. As expected, stimulation of quiescent 32P-labelled MCF-7 cells with 1 microM PMA resulted in the phosphorylation of a number of proteins. The PMA-induced phosphorylation of the proteins was abolished by preincubation of the cells with Ro 31-8220 (5 microM) for 20 min, or 10 microg/ml ET18-OCH3 for 3 h before stimulation with PMA. Thus under incubation conditions where ET18-OCH3 inhibited the proliferation of MCF-7 cells, the ether lipid potently inhibited the activity of PKC in intact cells. This inhibition was unlikely to be due to the effect of the compound on PKC translocation since there was little effect of ET18-OCH3 on the translocation of the alpha, gamma and epsilon species of PKC. These results suggest that a role for the inhibition of PKC activity by ET18-OCH3 in the mechanism of inhibition of cell proliferation by ET18-OCH3 cannot yet be discounted in epithelial cancer cells. In addition, we also observed that ET18-OCH3 enhanced the phosphorylation of selected proteins under basal unstimulated conditions. Although some of these proteins were also observed to be phosphorylated in response to PMA stimulation, the phosphorylation induced by ET18-OCH3 was not inhibited by Ro 31-8220, indicating that this was not mediated by PKC.

Biological Transport↗

PubMed1997

Determination of albuterol in plasma after aerosol inhalation by gas chromatography-mass spectrometry with selected-ion monitoring.

Albuterol is a beta2-adrenergic agonist commonly used as a bronchodilator for the treatment of patients with asthma. We have developed an assay to determine plasma levels as low as 50 pg/ml of albuterol by gas chromatography-mass spectrometry (GC-MS). This assay utilizes isotopically labeled albuterol ([13C]albuterol) as an internal standard. In this assay albuterol and the internal standard are recovered from 1 ml of plasma using solid-phase extraction. The samples are then derivatized to trimethylsilyl ethers using N,O-bis(trimethylsilyl)trifluoro-acetamide with 1% trimethylchlorosilane. The samples are then analyzed by GC-MS with selected-ion monitoring (SIM) for the ions m/z 369.15 and 370.15. The method has been validated for a concentration range of 50-10000 pg/ml in plasma.

Administration, Inhalation↗

PubMed1997

Effects of blocking B7-1 and B7-2 interactions during a type 2 in vivo immune response.

The costimulatory signal provided to T cells through CD28/CTLA-4 interactions is required for in vivo Th cell effector function associated with cytokine production. However, it is uncertain whether the two well-characterized ligands for these molecules, B7-1 and B7-2, differentially influence the consequent development of a type 1 or a type 2 primary response. We have examined the in vivo effects of blocking B7-1 and/or B7-2 ligand interactions on the type 2 mucosal immune response that follows oral infection of mice with the nematode parasite, Heligmosomoides polygyrus. Administration of the combination of anti-B7-1 and anti-B7-2 Abs inhibited H. polygyrus-induced increases in serum IgG1 and IgE levels, the expansion of mesenteric lymph node (MLN) germinal centers, in situ CD4+ T cell expansion, elevated blood eosinophils, and increased intestinal mucosal mast cells. Similarly, both Abs blocked MLN and Peyer's patch cytokine gene expression and elevations in MLN T cell-derived IL-4 protein secretion. However, in the same experiments, administration of either anti-B7-1 or anti-B7-2 Abs alone had little effect on any of these parameters. T cell and B cell activation was also blocked by the combination of anti-B7-2 and a B7-1-specific mutant Y100F CTLA-4Ig construct. These results suggest that to the extent that anti-B7-1 and anti-B7-2 mAbs block B7 interactions, either B7-1 or B7-2 ligand interactions can provide the required costimulatory signals that lead to T cell effector function during a type 2 in vivo immune response.

Animals↗