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Biomedical subjects

M Yang

Publications and source records attributed to M Yang.

At least 181 records · Page 10Linked to original sources
PubMed2000

[Hematopoietic stem cell transplantation for beta-thalassemia major].

OBJECTIVE: To discuss the feasibility of hematopoietic stem cell transplantation for beta-thalassemia major. METHODS: Bone marrow or cord blood of sibling was transplanted to 4 children with thalassemia, whose type of gene mutations was homozygous. All of them were diagnosed as beta-thalassemia major. HLA type was matched in 2 cases, and mismatched with 1 locus in the other 2. RBC type was mismatched in 1 cases. Pretransplant condition including busulfan (BU) 16 mg/kg, cyclophosphamide (CY) 200 mg/kg and anti-thymocyte globulin (ATG) 90 mg/kg. Graft-versus-host disease (GVHD) prophylaxis was cyclosporine-A (Cs-A) and methotrexate (MTX). RESULTS: Two Children had "serum sickness" during pretransplant condition; two children had grade II acute GVHD and 1 developed chronic GVHD, 1 case veno-occlusive disease (VOD), 1 case cytomegalovirus (CMV) interstitial pneumonia and 1 case fungemia post-transplant, which had been cured. The average time for peripheral WBC recovery > 1.0 x 10(9)/L was 16 days, 18 days for neutrophile granulocyte > 0.5 x 10(9)/L, 59 days for Plt > 50 x 10(9)/L and 39 days for Hb > 100 g/L. The average time for WBC recovery to normal was at day 24. The average time of last blood transfusion for patients was 29 days. The patients had to receive about 100 ml blood infusion per 2 months pretransplant, while not infused for 7 approximately 15 months up to now with Hb > 100 g/L. Gene mutation type of patients had changed to donor's. CONCLUSION: We report a successful transplantation of bone marrow and cord blood stem cells for thalassemia major in China. This will give a new strategy for treatment of the disease and further expand the use of stem cells transplantation.

Child, Preschool↗
PubMed2000

[Dural arterovenous fistula involving cavernous sinus].

OBJECTIVE: To study the pathogenesis and treatment of dural arteriovenous fistulas (DAVFS) involving the cavernous sinus. METHODS: 32 cases were embolized with particle by microcatheterization via endovascular approach and ervation by digital substraction angiography. Of the 32 cases, 14 were embolized by microsoils through superior petrosal sinus approach into the cavernous sinus. RESULTS: In 28 of the 32 cases the fistulas were completely embolized angiographically. The fistulas were partly embolized in 4 of the 32 cases. The carotid artery was compressed for 6 months. One week later, the fistula disappeared angiographically. No fistulas were found during the follow-up for 6 months to 8 years. CONCLUSION: Endovascular treatment of DAVFS involving the cavernous sinus is effective.

Adolescent↗
PubMed2000

[Studies on micronuclei induced by colchicine and cyclophosphamide using multicolor fluorescence in situ hybridization].

OBJECTIVE: To study chromosomal composition of micronuclei (MN) induced by colchicine (COL) and cyclophosphamide (CP) in mouse bone marrow erythrocytes. METHODS: Multicolor fluorescence in situ hybridization (FISH) with centromeric and telomeric DNA probes was applied to analyze chromosomal composition of micronuclei induced by COL and CP in mouse bone marrow erythrocytes. RESULTS: About 83.5% of COL-induced MN revealed both centromeric and telomeric signals. Of the CP-induced MN, 74.5% showed telomeric signals only. CONCLUSION: Majority of COL-induced MN contain whole chromosome and that of CP-induced MN mainly contain acentric fragments. Multicolor FISH with centromeric and telomeric DNA probes was a precise technique for analyzing chromosomal composition of MN.

Animals↗
PubMed2000

[Protective effect of bajitian oligosaccharide on PC12 cells lesioned by corticosterone].

OBJECTIVE: To study the possible mechanism of antidepressant effect of Bajitian oligosaccharide (MW-97). METHODS AND RESULTS: After incubating with the PC12 cells in the presence of corticosterone(2 x 10(-4) mol.L-1) for 48 h, it was found that MW-97 could protect PC12 cells from the lesion done by corticosterone in a concentration-dependent manner. Furthermore, using RT-PCR mediated cross-species partial cDNA cloning, it was found that MW-97 and desipramine(DIM) increased NGF, BDNF mRNA in the frontal cortex after chronic administration for 21 days, meanwhile, BDNF mRNA in hippocampus was also increased. CONCLUSION: The cytoprotective effect of MW-97 consists in the possible mechanism of its antidepressant action, while the increasing expression of neurotrophic factors (NGF, BDNF mRNA) may be contributes to the cytoprotective effect.

Animals↗
PubMed2000

[The neutralizing effect of histidine-rich-polypeptides on LPS from suspected periodontal pathogens].

OBJECTIVE: Human parotid histidine-rich-polypeptides (HRPs) are a family of lowmolecular-weight, cationic polypeptides. HRP-1, HRP-3 and HRP-5 comprise 85%-90% of the total HRPs and are called major HRPs. There were many researches indicating the antimicrobial activities of HRPs. Recently, HRPs were reported to have an inhibitory action on the lipopolysaccharide (LPS) of E. coli, with HRP-5 being the most effective one among the three major HRPs. Since the LPS of oral gram-negative bacteria is thought to be one of the important etiological factors during the development of periodontal diseases, our experiment was aimed to investigate the neutralizing effect of human parotid HRP-5 on the LPS of anaerobic suspected periodontal pathogens, which have different chemical structures and biological activities compared with LPS of aerobic E. coli. METHODS: By using of preparative acid urea polyacrylamide gel electrophoresis (AU-PAGE), HRP-5 was purified from parotid saliva collected from healthy adults. Two stains of suspected periodontal pathogens, Porphyromonas gingivalis (P.g, 47-A) and Fusobacterium nuceatum (F. n, separated from subgingival plaque of a patient with adult periodontitis) were mass cultured. After harvested in the midlogarithmic phase, they were washed and lyophilized. The LPS of dried bacteria were extracted by the modified Westphal hot phenol-water procedures and purified by the enzyme digestion plus ultracentrifugation. Limulus test was applied to test the neutralizing effect of HRP-5 on the LPS-induced gelation of Limulus amoebocyte lysate. In brief, the standard LPS of E. coli, or extracted LPS of P. g or F. n, was preincubated with HRP-5 in a tube separately in room temperature for 10 minutes. Then the reagent of Limulus amoebocyte lysate was added in all the tubes, continued the incubation in 37 degrees C for one hour. After that, the gelation level of every tube was observed. RESULTS: LPS extracted from P. g and F. n both showed good purity and strong activities to induce gelation of Limulus amoebocyte lysate. The gelation induced by LPS (1 ng/ml) of these two anaerobic suspected periodontal pathogens were weakly inhibited by HRP-5 (10 micrograms/ml), similar to that observed with standard LPS of E. coli. To get a complete neutralizing effect on LPS, it may be important to increase the concentration of HRP-5. CONCLUSION: It appeared that HRPs could neutralize the endotoxic properties of LPS of suspected periodontal pathogens, therefore may contribute to periodontal health. The present investigation further confirmed that HRPs are important components of the host non-immune defense system.

Escherichia coli↗
PubMed2000

[Distribution of IL-1 beta in periodontium of experimental osteoporosis rats during orthodontic tooth movement].

OBJECTIVE: The purpose of this research is to study the expression change of IL-1 beta induced by forces in the periodontium of osteoporosis rats. METHODS: Eighteen SD rats were randomly divided into three groups: the normal controlled, the normal orthodontically controlled and the experimental group of osteoporosis. Six weeks after ovariectomy, the first molars were moved mesially by 50 g force created from elastic-thread for one week in the force loading groups. Then the rats were killed, and the 5 microns-thick mesiodistal sections of maxillary teeth, including the roots and periodontium were made. IL-1 beta were localized immunohistochemically in periodontium, and semiquantitative analysis of cellular-staining intensity was done by microphotometry. RESULTS: The IL-1 beta of periodontal ligament cells without loading force were mildly stained in the controlled group, but the staining intensity increased significantly in the periodontium of orthodontically moved teeth, and it showed more strengthened staining in the periodontium of force-loading teeth in osteoporosis rats. CONCLUSION: The results demonstrate that periodontal ligament cells respond to mechanical force by increased production of IL-1 beta, and this can be enhanced by osteoporosis. IL-1 beta is an important cytokine in the process of orthodontically periodontal remodeling.

Animals↗
PubMed1999

Identification of modified tryptophan residues in apolipoprotein B-100 derived from copper ion-oxidized low-density lipoprotein.

Oxidative modifications of low-density lipoproteins (LDL) may contribute to the pathogenesis of atherosclerosis. Although the oxidation products of the lipid components of LDL have been studied extensively, less is known about the oxidation products of the apoprotein, apolipoprotein B-100. To identify the specific oxidative modifications, we oxidized LDL in the presence of Cu(2+), treated with DNPH, precipitated and delipidated the protein, digested the protein with trypsin, and analyzed the peptides by high-performance liquid chromatography. We isolated nine peptides that exhibited measurable absorbance at 365 nm, which is characteristic of hydrazones derived from DNPH and is not observed in peptides derived from unoxidized LDL. Unexpectedly, we obtained the same peptides with absorbance at 365 nm in Cu(2+)-oxidized LDL not treated with DNPH. N-terminal sequence analyses and mass spectrometry indicated that the peptides isolated from the Cu(2+)-oxidized LDL all contained kynurenine residues in place of Trp residues found in the native apoprotein. The product profile we observed in Cu(2+)-oxidized LDL was remarkably different from the profiles observed in LDL oxidized by HOCl or myeloperoxidase in vitro, and the preferential oxidation of Trp to kynurenine in Cu(2+)-catalyzed oxidation of LDL contrasts with the products observed following oxidation of LDL with HOCl or myeloperoxidase. Our studies to date support the working hypothesis that the specific products of protein oxidation are sufficiently distinct to be developed as biomarkers of proposed mechanisms of oxidation of LDL and biological molecules in other toxicities and diseases.

Amino Acid Sequence↗
PubMed1999

Assay of 2-naphthol in human urine by high-performance liquid chromatography.

This paper describes a novel liquid chromatographic method for the quantitation of 2-naphthol in human urine. Urine samples were extracted after enzymatic hydrolysis of glucuronides and sulfates; 2-naphthol was then separated using reversed-phase high-performance liquid chromatography. The corresponding detection limits were 0.04 ng/ml for the standard sample in acetonitrile and 0.13 ng/ml for urine samples. The level of urinary 2-naphthol in 100 Korean shipyard workers was analyzed using this new method. The level ranged from 0.21 ng/ml (0.26 micromol/mol creatinine) to 34.19 ng/ml (59.11 micromol/mol creatinine), and the mean+/-standard deviation was 5.08 ng/ml (6.60 micromol/mol creatinine)+/-5.75 ng/ml (9.22 micromol/mol creatinine). The mean+/-standard deviation of urinary 2-naphthol level of smokers, 7.03 ng/ml (8.49 micromol/mol creatinine)+/-6.16 ng/ml (10.23 micromol/mol creatinine), was significantly higher than that of non-smokers, 2.49 ng/ml (4.10 micromol/mol creatinine)+/-3.92 ng/ml (7.03 micromol/mol creatinine).

Acetonitriles↗
PubMed1999

Genetic analysis of the Drosophila ellipsoid body neuropil: organization and development of the central complex.

The central complex is an important center for higher-order brain function in insects. It is an intricate neuropil composed of four substructures. Each substructure contains repeated neuronal elements which are connected by processes such that topography is maintained. Although the neuronal architecture has been described in several insects and the behavioral role investigated in various experiments, the exact function of this neuropil has proven elusive. To describe the architecture of the central complex, we study 15 enhancer-trap lines that label various ellipsoid body neuron types. We find evidence for restriction of gene expression that is correlated with specific neuronal types: such correlations suggest functional classifications as well. We show that some enhancer-trap patterns reveal a single ellipsoid body neuron type, while others label multiple types. We describe the development of the ellipsoid body neuropil in wild-type animals and propose developmental mechanisms based on animals displaying structural mutations of this neuropil. The experiments performed here demonstrate the degree of resolution possible from the analysis of enhancer-trap lines and form a useful library of tools for future structure/function studies of the ellipsoid body.

Animal Structures↗
PubMed1999

Morphological, immunohistochemical and quantitative studies of murine brain mast cells after mating.

The mast cell is one of the immune cells, and can be triggered behaviorally to increase in the CNS of the sexually active dove. In the present study, we used ICR mice to investigate the number of brain mast cells in mated (one male with three female mice), non-mated (housed with female mice, but no mating) and control (four male mice housed together in one cage) male mice. We found that at least 40% of mated male mice had significant more mast cells than the maximum value seen in the controls, and that a significant correlation existed between the distribution index of mast cells and the postcoitum date. These mast cells were especially numerous in the thalamus and velum interpositum (VIP). Morphological observations showed that the increased mast cells were ultrastructurally similar to those in the controls, and displayed gonadotropin-releasing hormone (GnRH)-like immunoreactivity. Based on the facts that the number of brain mast cells in the male mice increased significantly after mating and that the change in the distribution of mast cells in the VIP and the thalamic parenchyma correlated well with time postcoitum, we speculate that, after mating, mast cells may migrate from the VIP to the thalamic parenchyma along the vascular tree of the brain. These results strongly suggest that mast cells are involved in the interaction among the immune, endocrine, and nervous systems in the mated male mouse brain.

Animals↗
PubMed1999

The Arabidopsis SKP1-LIKE1 gene is essential for male meiosis and may control homologue separation.

The yeast and human SKP1 genes regulate the mitotic cell cycle but are not yet known to be required for meiosis. Nine Arabidopsis SKP1 homologues have been uncovered and are named ASK1 through ASK9. Here, we report the isolation and characterization of a male sterile Arabidopsis mutant and show that the mutant defect was caused by a Ds transposon insertion into the ASK1 gene. In the ask1-1 mutant, abnormal microspores exhibit a range of sizes. Furthermore, during mutant male meiosis, although homologous chromosome pairing appeared normal at metaphase I, chromosome segregation at anaphase I is unequal, and some chromosomes are abnormally extended. Therefore, in ask1-1, at least some homologues remain associated after metaphase I. In addition, immunofluorescence microscopy indicates that the mutant spindle morphology at both metaphase I and early anaphase I is normal; thus, the abnormal chromosome segregation is not likely caused by a spindle defect. Because the yeast Skp1p is required for targeting specific proteins for ubiquitin-mediated proteolysis, we propose that ASK1 controls homologue separation by degrading or otherwise removing a protein that is required directly or indirectly for homologue association before anaphase I.

Amino Acid Sequence↗
PubMed1999

JAZ requires the double-stranded RNA-binding zinc finger motifs for nuclear localization.

We have cloned and characterized a novel zinc finger protein, termed JAZ. JAZ contains four C(2)H(2)-type zinc finger motifs that are connected by long (28-38) amino acid linker sequences. JAZ is expressed in all tissues tested and localizes in the nucleus, primarily the nucleolus. JAZ preferentially binds to double-stranded (ds) RNA or RNA/DNA hybrids rather than DNA. Mutation of individual zinc finger motifs reveals that the zinc finger domains are not only essential for dsRNA binding but are also required for its nucleolar localization, which demonstrates a complex trafficking mechanism dependent on the nucleic acid-binding capability of the protein. Furthermore, forced expression of JAZ potently induces apoptosis in murine fibroblast cells. Thus, JAZ may belong to a class of zinc finger proteins that features dsRNA binding and may regulate cell growth via the unique dsRNA binding properties.

3T3 Cells↗
PubMed1999

The peculiar nature of the guanidine hydrochloride-induced two-state denaturation of staphylococcal nuclease: a calorimetric study.

This work determines the ratio of DeltaH(vH) /DeltaH(cal) for staphylococcal nuclease (SN) denaturation in guanidine hydrochloride (GdnHCl) to test whether GdnHCl-induced denaturation is two-state. Heats of mixing of SN as a function of [GdnHCl] were determined at pH 7.0 and 25 degrees C. The resulting plot of DeltaH(mix) vs [GdnHCl] exhibits a sigmoid shaped curve with linear pre- and post-denaturational base lines. Extending the pre- and post-denaturational lines to zero [GdnHCl] gives a calorimetric DeltaH (DeltaH(cal)) of 24.1 +/- 1.0 kcal/mol, for SN denaturation in the limit of zero GdnHCl concentration. Guanidine hydrochloride-induced denaturation Gibbs energy changes in the limit of zero denaturant concentration (DeltaG degrees (N)(-)(D)) at pH 7. 0 were determined for SN from fluorescence measurements at fixed temperatures over the range from 15 to 35 degrees C. Analysis of the resulting temperature-dependent DeltaG degrees (N)(-)(D) data defines a van't Hoff denaturation enthalpy change (DeltaH(vH)) of 26. 4 +/- 2.8 kcal/mol. The model-dependent van't Hoff DeltaH(vH) divided by the model-independent DeltaH(cal) gives a ratio of 1.1 +/- 0.1 for DeltaH(vH)/DeltaH(cal), a result that rules out the presence of thermodynamically important intermediate states in the GdnHCl-induced denaturation of SN. The likelihood that GdnHCl-induced SN denaturation involves a special type of two-state denaturation, known as a variable two-state process, is discussed in terms of the thermodynamic implications of the process.

Calorimetry↗
PubMed1999

The significance of the homozygous CYP2A6 deletion on nicotine metabolism: a new genotyping method of CYP2A6 using a single PCR-RFLP.

A convenient and specific CYP2A6 genotyping method was developed in this study. This method consisting of a single PCR-RFLP is capable of resolving the genotype into either CYP2A6*1 (wild type), CYP2A6*2, or CYP2A6*3. Among 252 Japanese persons genotyped, 241 were genotyped as the wild type, 1 as an unknown variant, and none as either CYP2A6*2 or CYP2A6*3. A homozygous deletion was found in the 10 remaining subjects. To clarify the metabolic significance of this deletion in the whole human body, urinary cotinine, the principal metabolite of nicotine, was analyzed subsequent to smoking. Cumulated urinary cotinine excretion in the homozygously CYP2A6-deleted individuals was about one-seventh compared to the control group (wild type). This study provides a firm experimental basis for correlating genotypic characterization of CYP2A6 with phenotypic expression of nicotine metabolism.

Alleles↗
PubMed1999

Structure-activity relationships of a novel class of Src SH2 inhibitors.

The structure-activity relationships (SAR) of a novel class of Src SH2 inhibitors are described. Variation at the pY+1 and pY+3 side chain positions using 2,4- and 2,5-substituted thiazoles and 1,2,4-oxadiazoles as scaffolds resulted in inhibitors that bound as well as the standard tetrapeptide Ac-pYEEI-NH2.

Structure-Activity Relationship↗
PubMed1999

Characterization of transsynaptic tracing with central application of pseudorabies virus.

Although transsynaptic tracing with peripheral injection of pseudorabies virus (PRV) has been extensively characterized, several methodological issues related to central application of this tracer have not been addressed. In the present study, we addressed the following three issues by using microinjection of a cocktail containing PRV (Bartha strain) and cholera toxin subunit B (CTb) into different sites in the rat brain. First, we estimated PRV diffusion by examining injection sites at different times after application. Second, we tested whether PRV is taken up by fibers of passage following injections into the olivocerebellar pathway. Third, we developed criteria for leakage of PRV into cerebral ventricles. Our data indicate that (i) centrally injected PRV diffuses very little and produces focal injection sites; (ii) PRV is taken up and transported by fibers of passage, although less prominently than found for Ctb; (iii) PRV produces specific and easily identifiable ependymal cell as well as neuronal labeling following ventricular injection. This labeling can be used as a criterion for determining if labeling obtained was due to injected tracer leaking into brain ventricles. In summary, the present study provides new and important information about using PRV to trace central multisynaptic circuitry.

Animals↗
PubMed1999

Biosensor measurement of the interaction kinetics between insulin-like growth factors and their binding proteins.

The binding kinetics of human insulin-like growth factor binding protein (IGFBP) 1-6 for recombinant human insulin-like growth factor (IGF) I and II were measured and compared in the present study using surface plasmon resonance biosensor technique. Different concentrations of IGFBPs (5-100 nM) were allowed to interact with the immobilized IGF-I or IGF-II on sensor chip surface. Both des(1-3)IGF-I and insulin are known to bind weakly to the IGFBPs and therefore are used as negative controls for the binding experiments. The resultant sensorgrams were analyzed by using simple 1:1 binding model to derive both the association rate (k(a)) and dissociation rate (k(d)) constants for IGFBP-IGF interactions. The k(a) values of IGFBPs are in the range of 1x10(4) to 9x10(5) M(-1) s(-1) for IGF-I and 7x10(3) to 1.7x10(6) M(-1) s(-1) for IGF-II, respectively. The orders of k(a) for both IGF-I and IGF-II are IGFBP-3>IGFBP-5>IGFBP-6>IGFBP-4>IGFBP-2>++ +IGFBP-1. The k(d) values of IGFBPs are in the range of 1.5x10(-5) to 2x10(-4) s(-1) for IGF-I and 3.6x10(-5) to 3.7x10(-4) s(-1) for IGF-II, respectively. The order of k(d) for IGF-I is IGFBP-6>IGFBP-5>IGFBP-4>IGFBP-3>IGFBP-2>++ +IGFBP-1 and that for IGF-II is IGFBP-5>IGFBP-6>IGFBP-2>IGFBP-4>IGFBP-3>++ +IGFBP-1, respectively. The equilibrium affinity constants (K(A)) were calculated based on the ratio of k(a)/k(d) and were more precise than the published literature values based on competitive radioligand binding assays. The systematic study enables a direct comparison on the IGF-binding properties among the various IGFBPs, and the kinetic data provide additional information to delineate the physiological role of different IGFBPs in vivo.

Biosensing Techniques↗