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Biomedical subjects

X Yang

Publications and source records attributed to X Yang.

At least 793 records · Page 44Linked to original sources

[In vitro effects of hematopoietic growth factors on CD34+ cell from patients with myelodysplastic syndromes].

OBJECTIVE: To investigate the role of recombinant human hematopoietic growth factors (HGFs) in the treatment of myelodysplastic syndromes (MDS) patients. METHODS: CD34+ cells from 16 MDS patients were purified up to 96% approximately 99% by means of a immunomagnetic beads sorting system. The effects of individual and combination of HGFs on the proliferation and differentiation of the CD34+ cells were studied in semi -solid cultures. RESULTS: CD34+ cells were impaired and none of the individual HGF (GM-CSF, IL-3, SCF) could effectively support their proliferation and differentiation. rh-SCF combined with GM-CSF, IL-3 or Epo could partly increase the colony forming capacity of MDS hematopoietic progenitor, however, an increase of undifferentiated blast cells colonies occurred in some high-risk MDS patients at the same time. CONCLUSION: Proper combination of HGFs was essential for improving hematopoiesis in MDS patients.

Animals↗

[Lasting expression of hF IX mediated by myoblasts in mice].

OBJECTIVE: To develop experimentally gene therapy for hemophilia B by using C2C12 cells and C3H mice as models. METHODS: The retroviral vector containing MCK enhancer and hCMV promoter was constructed and was used to transfect PA317 cells. Then the viral particles were used to transfect C2C12 cells and select the G418r clones,which were further confirmed by Southern, Northern, Western blots. And then the C2C12 cells expressing hF IX protein were injected into C3H mice and hF IX expression in vivo was studied. RESULTS: The expression level of hF IX in vivo was positively correlated with the number of injected cells, and the in vitro expression level of these cell clones. The expression of hF IX could be enhanced and prolonged by immunosuppressive agents. CONCLUSION: Allogeneic myoblasts transplantation leads to immunorejection, but repeated injections at a longer intervals could improve the existing rate of transplanted cells. And this might be helpful to gain insight into the myoblast-mediated gene therapy for hemophilia B patients.

Animals↗

[Expression of ferritin receptor on placental microvillous membrane and its role in maternal-fetal iron transport].

OBJECTIVE: In order to investigate the expression of ferritin receptor (FnR) on human placental microvillous membrane in mid-pregnancy and its implication. METHODS: Radioligand binding assay was used, with horse spleen 125I-apo-ferritin as radioligand. RESULTS: The FnR expression in normal placental microvillous membrane in mid-pregnancy was (9.63 +/- 4.72) x 10(12) sites/mg pro, similar to that in normal placental of term delivery,and the dissociation constants (Kd) between them had also no significant difference: Competitive binding assay indicated that the receptor bound basic ferritin with higher affinity than acidic one. CONCLUSIONS: The expression of FnR in mid-pregnancy almost come to mature level; The FnR on placenta might be involved in maternal-fetal iron transport, transfering iron from mother to fetal.

Apoferritins↗

[Localization of ferritin in placental villous tissue and its role in the maternal-fetal iron transport].

OBJECTIVE: To investigate the localization and quantity of ferritin in placental villous tissue and its implications. METHODS: The localization of ferritin in placental villous tissue at different gestation period was assayed by immunocytochemistry. The quantity of ferritin in placental villous tissue was determined with radioimmunoassay. RESULTS: Ferritin was localized in all layers of the trophoblast, especially in the surface of the syncytiotrophoblast and basal membrane. The quantity of ferritin in placental villous tissue was increased with gestation progress and decreased in pregnant women with IDA. CONCLUSIONS: Ferritin may be the important carrier of transplacental iron transport, and its receptor plays a crucial role in the course. The placental iron transport varied with gestation progress and iron nutritional status of the mother.

Anemia, Iron-Deficiency↗

[Liquid chromatographic separation of pepsinogens].

In this work, we reported a method for the purification of pepsinogens from human gastric mucosa with high pressure gel filtration chromatography (HPLC) and medium pressure anion exchange chromatography (MPLC). First, the two fractions of Pg1 and mixture of PGI and PGII were separated from pepsinogens with HPLC on a Bio-Sil SEC-125 column within 50min. Then the mixture of fractions of PGI and PGII was further separated with MPLC on a Bio-Scale Q2 column within 30min.

Chromatography, High Pressure Liquid↗

[The development of a gas chromatograph used in gas analysis below atmospheric pressure].

During the process of setting up a carbon monoxide infrared laser generator, it's necessary to analyze the gaseous components produced in laser cavity under reduced pressure for understanding its charging regularity. In this paper a self-made gas chromatograph for analysis of some special gas mixtures is described. This instrument can analyze O2, N2, CO (with normal contents) and trace CO2 simultaneously below atmospheric pressure. The gas sample below atmospheric pressure was taken into a small piston cylinder and then compressed by pushing the piston for injection. The injected sample was carried by He into two parallel chromatographic columns and detected separately. The gases with normal contents were detected by a thermalconductivity detector (TCD) and the trace CO2 was detected by a He ionization detector (HeID). The detection sensitivity of CO2 is in the order of magnitude of 10(-6) V/V. The experimental results show that this instrument with advantages of simple design, easy operation and good stability is suitable for on-line analysis of gas samples below atmospheric pressure. We have successfully analyzed on-line the gaseous components produced in the laser cavity by using this equipment. The technique described here can be used for complete analysis of gas samples below atmospheric pressure in many other fields.

English Abstract↗

[Studies on polysaccharide peptide with high performance gel permeation chromatography].

High performance gel permeation chromatography (HPGPC) has been widely used in the fraction and molecular-weight (MW) estimation of polysaccharides. In this paper, we estimated the MW of major component of polysaccharide-peptide from Corilous versicolor (PSP). These three-components were separated by using two column system (TSK-40 and TSK-50 gel columns) and different NaCl concentration in mobile phase. Calibration curve was prepared from the known MW Dextran T-system. It was found that the MW of PSP was changed in accordance with ion concentration of mobile phase. The MWs of the main component are more than 500000, 120000-19000 and less than 10000 respectively as the NaCl concentrations in mobile phase were less than 0.002, 0.006-0.04 and more than 0.06mol/L. The results showed that the MW of polysaccharide-peptide determined by HPGPC is a relative value that is changed with the concentration of mobile phase. The reason is that the molecular configuration and size of polysaccharide-peptide are changed along with electrolyte concentration in mobile phase. That is, the lower the electrolyte concentration is, the larger the molecular size of polysaccharide-peptide is. We suggest that it is still useful for quality control of PSP as the mobile phase is fixed.

Chromatography, Gel↗

[Selection of TLC solvent system for separating substituted ferrocyclopentadienyl borane using orthogonal array method].

A series of substituted ferro-cyclopentadienyl borane compounds were separated by using thin-layer chromatographic technique. The optimum mobile phase was acetone-cyclohexane-tetraethyl ammonium halide (0.59:8.24:1.18). The orthogonal array method was used to get the optimum chromatographic condition. The types and the concentrations of different kinds of ion-pair reagents, the contents of acetone and cyclohexane were considered as factors which affected the results of separation. The operating conditions for each experiment was adopted based on the L9 (3(4)) design. A statistical method of variance analysis was used to evaluate the overall degree of separation (D(S)), a criterion which reflects the degree of optimization. As a result, we found the most important factors that affected the separation and the optimum chromatographic condition for 8 compounds of this sort.

English Abstract↗

[Method of cyanogen bromide-fluorimetry determination of trace amount of selenomethionine in grain and blood].

Selenomethionine (SeMet) reacted with cyanogen bromide (BrCN) quantitatively forms CH3SeCN. After extracted with CHCl3, the Se of CH3SeCN is acid-digested to Se(IV). Then 2,3-diaminonaphthalene is used to determine the fluorescent Se value of 4,5-benzopiaselenol. The determination limit of this method was 3 ng/g SeMet. The accuracy of 10-500 ng Se in SeMet standard was 91.8%-97.6%. RSD was 1.9%-6.3%. Recoveries for grain and blood were 92.3%-96.7%. RSD was 2.7%-5.1%. The RSD for samples was 2.7%-9.0%. Selenocystine, selenocystiene selenite and methionine did not interfere with the determination.

Cyanogen Bromide↗

[Determination of Ag, Cu, Zn and Cd in silver brazing filler metals by ICP-AES].

A method of simultaneous and direct determination for Ag, Cu, Zn and Cd in silver brazing filler metals by ICP-AES is reported. The spectral interferences and effect of acidity have been investigated. Working conditions were optimized. The method has been applied to the analysis of silver brazing filler metals with RSD of 4-7% and recovery of 94-105%. This method was accurate, simple and rapid.

English Abstract↗

Estrus, fertility, early embryo development, and autologous embryo transfer in laboratory woodchucks (Marmota monax).

Reproductive parameters were studied in female laboratory woodchucks over a 6-year period. The pregnancy rate in adult females after ad libitum exposure to a male (n = 643) was 72%; that after limited (4 to 8 h) exposure with video observations (n = 31) was only 37%. However, limited exposure resulted in a 75% ovulation rate, suggesting a problem with fertilization and not ovulation. The ovulation rate was 0% in the absence of mating (n = 10), suggesting that spontaneous ovulation is not the usual situation in this species. With ad libitum mating, fertility was greater (P < 0.05) for 2- to 4-year-old females (73%) than for 1-year-old (56%) or > or = 5-year-old (58%) females. Mean (+/-SEM) litter size at birth was greater (P < 0.05) for 2- to 5-year-old animals (4.0 +/- 0.1) than for 1-year-old animals (2.9 +/- 0.2). However, in 1-year-old females the neonatal mortality was low, and the litter size at weaning (2.4 +/- 0.3 pups) did not differ from that of older females (2.7 +/- 0.1 pups). The measurement of vulval diameter every 3 to 4 days in 31 adult females suggested that enlargement to > or = 7 mm was indicative of proestrus or estrus and was a good indicator of willingness to accept mounting by males. Vulval swelling > or = 7 mm (n = 31) lasted 5 to 47 days. Fertile mating occurred from 3 to 30 days after vulval diameter reached > or = 7 mm, near the time of peak enlargement (10.1 +/- 0.2 mm), and was followed in several days by a decrease in vulval size. Fifty-three females were allowed only limited (6 to 10 h) video-observed exposure to males, with (n = 26) or without (n = 27) another 12 to 24 h of unobserved exposure, and were examined surgically for ovulation and pregnancy at known times after mating. There were no consistent differences among nonovulating, ovulating nonfertile, and fertile females in the longest duration of mating (7.2 +/- 0.4 min) or in the number of matings lasting > or = 1 min (5.7 +/- 0.8). Compared with nonfertile females, fertile females experienced more matings > or = 3 min in duration (3.3 +/- 0.4 versus 1.8 +/- 0.2) and > or = 5 min in duration (2.9 +/- 0.4 versus 1.1 +/- 0.1). At postmating days 1, 2, 3, 4, 5, 6, and 7 the embryos were observed to be 1-, 4-, 4- to 32-cell morulae, 16- to 60-cell morulae, free-floating blastocysts, anti-measometrial peri-implantation blastocysts, and attached blastocysts respectively. Autologous transfer of day-5 uterine embryos of normal appearance into the noncontiguous contralateral uterine horn was performed in five females that had been subjected to a hemi-ovariectomy. Transfers resulted in pregnancy and litters in four of the five females. The results suggest that this large, circannual sciurid rodent can be successfully and predictably bred as a laboratory animal model, has vulval changes indicative of proestrus, is principally an induced ovulator, has an early embryo development rate similar to that of rats and mice, and may be amenable to reproductive technologies developed for more common laboratory rodents.

Animals↗

Elements between the protein-coding regions of the adjacent beta 4 and alpha 3 acetylcholine receptor genes direct neuron-specific expression in the central nervous system.

The expression patterns of three clustered neuronal nicotinic acetylcholine receptor (nAchR) subunit genes ordered beta 4, alpha 3, and alpha 5 overlap extensively in the peripheral nervous system (PNS) but only partially in the central nervous system (CNS). We have begun to investigate cell type-specific cis elements regulating these genes by analyzing in both cell culture and transgenic mice, a 2.8-kb fragment (-2732/+47) containing the alpha 3 promoter region, the beta 4/alpha 3 intergenic region, and a portion of the beta 4 3'-untranslated exon. The -2732/+47 fragment is preferentially active in PC12 cells relative to nonneural cell lines. Deletion analysis revealed a cell type-specific positive transcriptional element positioned in the beta 4 3'-untranslated exon. The positive element is likely to be an enhancer and not a second alpha 3 promoter, because no alpha 3 exons are present in this region. Having shown in cell culture that cell-type specific cis elements are positioned between the beta 4 and alpha 3 coding regions, we investigated the activity of -2732/+47 in vivo. Transgenic mice were generated, which carry the lacZ gene fused downstream of -2732/+47. Expression of the lacZ transgene is restricted to neurons of the CNS; no expression was detected in the PNS or in nonneural tissues. LacZ-positive cells were detected virtually exclusively in a subset of CNS nuclei that transcribe the endogenous alpha 3 gene. Some overlap was seen with the beta 4 gene, but nearly none with the alpha 5 gene. Our results demonstrate that cis elements positioned between the alpha 3 and beta 4 coding regions are important for establishing part of the restricted CNS patterns of beta 4, alpha 3, and alpha 5 gene transcription.

Animals↗

Oxidation of phenothiazines by human term placental peroxidase in non-smokers.

Several phenothiazine derivatives have been shown to cause reproductive toxicity. The biochemical mechanisms responsible for these effects are not fully understood at present. In this study, we investigated hydrogen peroxide-dependent oxidation of seven phenothiazines by purified human term placental peroxidase. Promazine was employed as the prototype phenothiazine drug. Promazine was easily oxidized to a cation radical (absorption maximum at 513 nm) when incubated under acidic conditions with human term placental peroxidase in the presence of hydrogen peroxide. The rate of the radical formation was 350 +/- 30 nmol/min/mg protein when 30 micrograms of the purified peroxidase was incubated with 10 mM promazine and 1.0 mM hydrogen peroxide in acetate buffer pH 3.5. The reaction was linear with respect to time, exhibited dependence on the amount of enzyme, and the concentration of promazine and hydrogen peroxide. Collectively, the results suggest an enzymatic nature of the reaction. Reduced glutathione, ascorbate, and NADH suppressed the rate of promazine radical formation presumably by its reduction back to promazine in a concentration-dependent manner. Concomitantly, NADH was co-oxidized. The cation radical formation and NADH oxidation declined significantly by potassium cyanide and sodium azide, the known peroxidase inhibitors. The enzyme also oxidized chlorpromazine, triflupromazine, trifluoperazine, trimeprazine, fluphenazine, and perphenazine in the presence of hydrogen peroxide. The evidence gathered in this study suggests that peroxidative bioactivation of phenothiazines to their corresponding cation radical species by the human placental peroxidase may represent one of the biochemical mechanisms responsible for the reported developmental toxicity of these chemicals.

Antipsychotic Agents↗

Change of level and expression of endothelin-1 in the lungs of rats with hypoxic pulmonary hypertension.

OBJECTIVE: To determine whether mRNA for endothelin-1 was present and changed in the lungs of rats exposed to chronic hypoxia. METHODS: cRNA probes for rat ET-1 and c-fos were labelled by in vitro transcription with digoxigenin-UTP for in situ hybridization. Endothelin-1 like immunoreactivity (ET-1-LI) in plasma as well as in lung homogenate was measured by radioimmunoassay. RESULTS: The vascular endothelial cells of pulmonary arteries and the bronchiolar epithelial cells were strongly positive for the ET-1 probe. The perivascular and peribronchiolar smooth muscle were positive for the c-fos probe, primarily. There was little expression of ET-1 and c-fos in the rat lungs in controls. There was significant increase of ET-1-LI levels in rats exposed to hypoxia for 3 weeks, the concentration ET-1-LI in venous samples was 3.85 +/- 1.52 ng/L (P < 0.05 as compared with the control groups), in arterial sample was 4.72 +/- 1.66 ng/L (P < 0.05) and in lung was 2.06 +/- 0.68 ng/g wet lung weight (P < 0.05). CONCLUSIONS: Chronic hypoxia elevated the mean pulmonary arterial pressure, caused vessel remodelling and the right ventricular hypertrophy. These changes were accompanied by an increase of ET-1 in plasma and lung homogenate. The expression and production of ET-1 were localized to endothelium and airway epithelium in the lungs.

Animals↗

Human cytomegalovirus matrix protein PP150 is efficiently presented as one of target antigens for cytotoxic T lymphocyte recognition.

OBJECTIVE: To determine whether human cytomegalovirus (CMV) matrix protein PP150 is efficiently presented for CMV-specific CTL recognition. METHODS: Recombinant vaccinia virus (Vac. PP150) encoding human CMV structural matrix protein PP150 was constructed with vaccinia vector PSC11 and it was used to stimulate peripheral blond mononuclear cells from 5 CMV seropositive individuals. RESULTS: PP150-specific CTLs could be generated in all of them, which not only lysed Vac. PP150-infected fibroblasts, but also lysed CMV-infected targets. In the presence of RNA synthesis inhibitor Actinomycin D (Act D) or at very early stage of infection, PP150-specific CTL lysed CMV-infected targets as efficiently as in the absence of Act D or at late stage of infection. CONCLUSIONS: PP150 exogenously introduced with the virus infection could be efficiently presented prior to viral DNA replication and PP150 is one of the major target antigens for CTL recognition.

Antigens, Viral↗

On the roles of inscuteable in asymmetric cell divisions in Drosophila.

Three processes requiring insc, protein localization, RNA localization, and mitotic spindle orientation, appear to be tightly correlated but nevertheless independent. The apical cortical localization of Insc protein occurs prior to mitosis and precedes the basal localization of Pros, Numb, and pros RNA. In insc null mutants, neuroblasts divide in more random orientations and fail to correctly localize Pros, Numb, and pros RNA to their correct position during mitosis. Our results suggest that asymmetric localization of Insc during interphase functions to establish positional information for spindle orientation and the basal localization of Numb, Pros, and pros RNA during mitosis. Insc probably interacts with a number of interacting partners in order to effect its various functions. One of its partners, Stau, is involved exclusively in the process of RNA localization.

Amino Acid Sequence↗

[A quantitative study on the cells of cochlear nucleus in rats at different ages].

The volume of cochlear nucleus and nucleolus, neural populations were determined in rats of three groups aged (1, 2-3, 22-28 months). The study data indicated that there is no difference between the volume of cochlear nucleus in the aged and adult animals (P > 0.05). Compared with the other two groups, the aged animals show a reduction of volume of necleolus of cochlear nucleus neurons (P < 0.01), a loss of neurons (spherical cell and multipolar cell) (P < 0.01). On the other hand, it shows an increase in number for the glial cells in the aged animals (48.48%, P < 0.01). Quantitative results of the cell of cochlear nucleus were analysed. We suggest that central changes also play an important role in presbycusis.

Age Factors↗

[Protective effect of magnesium on ionic channels in guinea pig ventricular myocytes during hypoxia].

OBJECTIVE: To determine effects of Mg2+ on potassium current, transient inward current (Iti) and Na(+)-Ca2+ exchange current in isolated ventricular myocytes during hypoxia. METHODS: Single myocytes were isolated from ventricles of adult guinea pig hearts. The patch clamp technique in whole cell configuration was used to study ionic currents, and experiments were performed in an experimental chamber that allows the cell to be exposed to a sufficiently low O2 pressure. RESULTS: The 10 mmol/L intracellular free Mg2+ (Mg2+i) had no effect on action potential abbreviation under normoxic condition, however, the action potential abbreviation during hypoxia was markedly inhibited in presence of 10 mmol/L Mg2+i. The amplitude of hypoxia-induced time independent outward K+ current was also greatly reduced when applied 10 mmol/L Mg2+ in pipette solution. In early reoxygenation, Iti occurred in 73% of myocytes, 10 mmol/L extracellular Mg2+ during hypoxia and reoxygenation lowered incidence of Iti, 10 mmol/L Mg2+ could partly inhibit the Na(+)-Ca2+ exchange current in normoxic condition. CONCLUSION: Intracellular Mg2+ could block hypoxia-induced outward current through ATP sensitive potassium channels. Mg2+i plays an important role in preventing depletion of cellular K+ and regulates beat-to-beat cardiac activity. Extracellular Mg2+ most probably decreases the signs of calcium overload in early reoxygenation by inhibiting Na(+)-Ca2+ exchange.

Animals↗