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Biomedical subjects

H Yu

Publications and source records attributed to H Yu.

At least 91 records · Page 5Linked to original sources

Development and applications of a complete set of rice telotrisomics.

We previously isolated a complete set of primary trisomics along with many other aneuploids from triploid plants derived from an indica rice variety "Zhongxian 3037." About 30,000 progeny from these trisomic and aneuploid plants were grown each year from 1994 to 1999. The variants that differed morphologically from both the diploids and the original primary trisomics were collected for cytological identification. From these variants, a complete set of telotrisomics covering all 24 rice chromosome arms was obtained. The identities of the extra chromosomes were further confirmed by dosage analysis of the RFLP markers on extra chromosome arms. The telocentric nature of the extra chromosomes in these stocks was verified by fluorescence in situ hybridization (FISH) using a rice centromeric BAC clone as a marker probe. In general, the shorter the extra chromosome arm of a telotrisomic, the stronger the resemblance it bears to the diploid; the longer the extra chromosome arm, the stronger the resemblance to the corresponding primary trisomic. We demonstrated that DNA clones can be rapidly assigned to specific chromosome arms by dosage analysis with the telotrisomics. We also showed that telotrisomics are valuable tools for chromosome microdissection and for developing chromosome-specific DNA markers.

Chromosomes↗

Genetic footprinting in bacteria.

In vivo genetic footprinting was developed in the yeast Saccharomyces cerevisiae to simultaneously assess the importance of thousands of genes for the fitness of the cell under any growth condition. We have developed in vivo genetic footprinting for Escherichia coli, a model bacterium and pathogen. We further demonstrate the utility of this technology for rapidly discovering genes that affect the fitness of E. coli under a variety of growth conditions. The definitive features of this system include a conditionally regulated Tn10 transposase with relaxed sequence specificity and a conditionally regulated replicon for the vector containing the transposase and mini-Tn10 transposon with an outwardly oriented promoter. This system results in a high frequency of randomly distributed transposon insertions, eliminating the need for the selection of a population containing transposon insertions, stringent suppression of transposon mutagenesis, and few polar effects. Successful footprints have been achieved for most genes longer than 400 bp, including genes located in operons. In addition, the ability of recombinant proteins to complement mutagenized hosts has been evaluated by genetic footprinting using a bacteriophage lambda transposon delivery system.

Bacteriophage lambda↗

Transduction of cellular sequence by a human immunodeficiency virus type 1-derived vector.

During studies examining the rate of human immunodeficiency virus type 1 (HIV-1) mutation in a single cycle of replication, the 5' long terminal repeat of one progeny provirus was found to contain an insertion of 147 bp including an entire tRNA sequence as well as an additional 66 bp insertion of nonviral origin. Database searches revealed that 65 of 66 bp aligned with the human CpG island sequence found on chromosomes 6, 14, and 17. Therefore it seems probable that it is of human cellular sequence origin and was transduced by HIV-1. This is the first demonstration that HIV-1 can capture a cellular sequence. The site of integration of the parental provirus was mapped to chromosome 1p32.1. Sequence with homology to the transduced CpG island was not found on chromosome 1, suggesting that the transduced cellular sequence was not linked to the site of viral integration.

Base Sequence↗

Nystatin and valinomycin induce tubuloglomerular feedback.

The macula densa expresses a luminal Na(+)-K(+)-2Cl(-) cotransporter and a basolateral Cl(-) conductance. Although it is known that cotransport of Na(+), K(+), and Cl(-) is the first step in tubuloglomerular feedback (TGF), subsequent steps are unclear. We hypothesized that Na(+)-K(+)-2Cl(-) entry via the luminal Na(+)-K(+)-2Cl(-) cotransporter elevates intracellular Cl(-), increases electrogenic Cl(-) efflux across the basolateral membrane, and depolarizes the macula densa, initiating TGF. We perfused afferent arterioles with macula densa attached. The macula densa was perfused with solutions containing either 5 mM Na(+) and 3 mM Cl(-) (low NaCl) or 80 mM Na(+) and 77 mM Cl(-) (high NaCl). When the macula densa perfusate was changed from low to high NaCl, afferent arteriole diameter decreased from 15.8 +/- 0.8 to 13.1 +/- 0.7 mm (P < 0.05). Adding 10 microM furosemide to the macula densa lumen blocked TGF. When nystatin, a group I cation ionophore, was added to the macula densa lumen together with furosemide in the presence of low NaCl, it induced TGF (from 18.0 +/- 1.5 to 15.6 +/- 1.6 mm; P = 0.003). When valinomycin, a K(+)-selective ionophore, was added to the macula densa lumen together with furosemide in the presence of low NaCl containing 5 mM K(+), it did not induce TGF. Subsequent addition of 50 mM KCl to the macula densa perfusate induced TGF (from 21.7 +/- 0.8 to 17.5 +/- 1.3 mm; P = 0.0047; n = 6). Adding 50 mM KCl without valinomycin did not induce TGF. When 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB; 1 microM), a Cl(-) channel blocker, was added to the bath, it blocked TGF induced by high NaCl, but did not block TGF induced by valinomycin plus 50 mM KCl. NPPB did not alter afferent arteriole constriction induced by norepinephrine. We concluded that increased NaCl in the lumen of the macula densa leads to influx of Cl(-) via the Na(+)-K(+)-2Cl(-) cotransporter. The accelerated transport increases intracellular Cl(-). The subsequent exit of Cl(-) across the basolateral membrane via Cl( -) channels in turn leads to depolarization of the macula densa and thereby induces TGF.

Animals↗

7-Dehydrocholesterol-dependent proteolysis of HMG-CoA reductase suppresses sterol biosynthesis in a mouse model of Smith-Lemli-Opitz/RSH syndrome.

Smith-Lemli-Opitz/RSH syndrome (SLOS), a relatively common birth-defect mental-retardation syndrome, is caused by mutations in DHCR7, whose product catalyzes an obligate step in cholesterol biosynthesis, the conversion of 7-dehydrocholesterol to cholesterol. A null mutation in the murine Dhcr7 causes an identical biochemical defect to that seen in SLOS, including markedly reduced tissue cholesterol and total sterol levels, and 30- to 40-fold elevated concentrations of 7-dehydrocholesterol. Prenatal lethality was not noted, but newborn homozygotes breathed with difficulty, did not suckle, and died soon after birth with immature lungs, enlarged bladders, and, frequently, cleft palates. Despite reduced sterol concentrations in Dhcr7(-/-) mice, mRNA levels for 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the rate-controlling enzyme for sterol biosynthesis, the LDL receptor, and SREBP-2 appeared neither elevated nor repressed. In contrast to mRNA, protein levels and activities of HMG-CoA reductase were markedly reduced. Consistent with this finding, 7-dehydrocholesterol accelerates proteolysis of HMG-CoA reductase while sparing other key proteins. These results demonstrate that in mice without Dhcr7 activity, accumulated 7-dehydrocholesterol suppresses sterol biosynthesis posttranslationally. This effect might exacerbate abnormal development in SLOS by increasing the fetal cholesterol deficiency.

Animals↗

Purification and characterization of ginsenoside-beta-glucosidase from ginseng.

The ginsenoside-beta-glucosidase that hydrolyzes the beta-(1-->2)-glucoside of the ginsenoside Rg3 sugar moiety to ginsenoside Rh2 was isolated from the ginseng root, and the enzyme was purified and characterized. The enzyme was purified to one spot in SDS polyacrylamide gel electrophoresis, and its molecular weight was about 59 kDa. The optimum temperature of the ginsenoside-beta-glucosidase was 60 degrees C, and the optimum pH was 5.0. Ca2+ ion had positive effect on ginsenoside-beta-glucosidase, while Cu2+ had negative effect on it. The ginsenoside-beta-glucosidase may be a special beta-glucosidase that is different from the original exocellulase such as beta-glucosidase (EC 3.2.1.21).

Chromatography, DEAE-Cellulose↗

Continuous positive airway pressure normalizes cardiac autonomic and hemodynamic responses to a laboratory stressor in apneic patients.

OBJECTIVES: We examined the effect of continuous positive airway pressure (CPAP) treatment for sleep apnea on cardiac contractility, heart rate variability, and hemodynamics at rest and in response to a laboratory stressor. SUBJECTS AND INSTRUMENTATION: Forty-one apneic patients were studied on three occasions: before treatment, after 1 full night of CPAP treatment, and after 1 week of CPAP treatment. The subjects were randomly assigned to receive effective treatment or placebo. Contractility and hemodynamics were determined with impedance cardiography, and parasympathetic activity was assessed by analysis of heart rate variability. Measures were determined at rest and in response to a stressor. DESIGN AND RESULTS: For the cardiac sympathetic (contractility) measures (preejection period, cardiac acceleration index [CAI], and low-frequency/high-frequency ratio) significant interactions were found in the combination treatment (CPAP vs placebo) by study day (day 1, day 3, day 11) by test period (baseline, preparation, talking) [p < 0.01]. For these measures, there were no differences between the treatment groups or responses to the stressor on day 1. Levels in placebo-treated subjects did not change or respond on the subsequent study days. In the CPAP-treated subjects, there was a decrease in these indexes at baseline, which became significantly lower by day 11 (ie, CAI levels were 24 Omega/s(2), 22 Omega/s(2), and 14 Omega/s(2) on day 1, day 3, and day 11, respectively). These measures also became responsive to the stressor by showing increased sympathetic activity (CAI levels on day 11 were 14 Omega/s(2) at baseline, 32 Omega/s(2) during speech preparation, and 36 Omega/s(2) while speaking). The parasympathetic indexes, such as high-frequency power or band of heart rate variability as determined by spectral analysis, showed a significant day-by-treatment interaction (p < 0.005), whereas the CPAP- treated group had significantly more parasympathetic activity after 1 week of treatment. For the hemodynamic measures (stroke volume [SV], cardiac output, and systemic vascular resistance [SVR]), there were significant treatment-by-study day-by-test-period interactions (p < 0.01). SV and cardiac output increased across days, and SVR decreased in the CPAP-treated patients. CONCLUSIONS: These results indicate that CPAP normalizes contractility, increases cardiac vagal tone, and changes hemodynamic regulation from being resistance dominated to being cardiac dominated. Thus, after 1 week of treatment with CPAP, many of the indicators of poor cardiac functioning in apnea patients are improved.

Adult↗

AMPA receptor function is altered in GLUR2-deficient mice.

The GluR2 subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid (AMPA) receptor determines many of the biophysical properties of native AMPA receptors, including Ca++ permeability. Genetically engineered mice unable to edit the Q to R site of the GluR2 subunit die within 3 wk postpartum, presumably due to toxicity associated with enhanced Ca++ influx through AMPA receptors. In contrast, disruption of the gene encoding GluR2 is not necessarily lethal. The objective of this study was to explore potential mechanisms that permit survival of GluR2 (-/-) mice despite AMPA receptors that are highly Ca++ permeable. Whole-cell, patch-clamp recording of AMPAreceptor responses in cortical pyramidal cells revealed that the kinetics of recovery from desensitization were significantly slower for receptors from GluR2 (-/-) mice compared to receptors from GluR2 (+/+) mice. The recovery time constants for AMPA receptors from GluR2 (-/-) and GluR2 (+/+) mice were 109.8 +/- 17 ms and 54.4 +/- 7.1 ms, respectively. The slower recovery kinetics would be expected to reduce Ca++ influx during repetitive stimulation. Because both RNA editing at the R/G site and alternative splicing of the flip and flop module affect AMPA receptor desensitization recovery rates, the possibility that these mechanisms were changed in GluR2 (-/-) mice was investigated. On a macroscopic level, neither editing nor splicing of the GluR-1, 3 or 4 subunits were changed in GluR2 (-/-) mice compared to GluR2 (+/+) mice. In summary, an increase in the time constant for recovery from desensitization may contribute to the ability of GluR2 (-/-) to survive.

Animals↗

Association of a novel point mutation (C159G) of the CTLA4 gene with type 1 diabetes in West Africans but not in Chinese.

Here, we report on the detection of a novel point mutation of the CTLA4 gene at nucleotide position 159 (C-->G) leading to amino acid substitution at position 53 (I-->M), as well as its association with type 1 diabetes in two ethnically distinct populations. Subjects included 182 unrelated type 1 diabetes children and 201 control subjects from Ghana, West Africa. The Chinese study population consisted of 350 type 1 diabetic children and 420 healthy control subjects from central China. Polymerase chain reaction-single-strand conformation polymorphism and sequence analysis were used to screen for polymorphisms in the CTLA4 gene. CTLA4 49 (A-->G) mutation conferred a risk of type 1 diabetes in the Chinese children (odds ratio 1.78, 95% CI 1.58-2.0), but not in the West African children (1.17, 0.84-1.64). On the other hand, the novel CTLA4 159 (C-->G) mutation conferred a risk of type 1 diabetes in the West African children (2.1, 1.54-2.86), but not in the Chinese type 1 diabetic children. The novel CTLA4 gene polymorphism at nucleotide position 159 significantly associated with type 1 diabetes in West Africans, but not in Chinese. On the other hand, the CTLA4 gene polymorphism at nucleotide position 49 significantly associated with type 1 diabetes in Chinese, but not in West Africans.

Abatacept↗

mRNA and protein expression of Fas associated death domain protein in apoptosic hepatocyte induced by tumor necrosis factor-alpha.

OBJECTIVE: To study the expression of protein and mRNA of Fas associated death domain protein (FADD) in the apoptosic hepatocyte induced by D-galactosamine (GalN) and TNF-alpha. METHODS: Fulminant hepatic failure (FHF) was induced by injection of GalN into sensitized BALB/c mice by TNF-alpha. Hepatocyte apoptosis was examined by electric microscopy, in site end labeling (ISEL) and DNA electrophoresis on 1.5% agarose gel. The expression of protein and mRNA of FADD were detected by immunohistochemistry and PT-PCR. RESULTS: GalN/TNF-alpha may induce hepatocyte apoptosis, necrosis and liver failure. The expression of protein and mRNA of FADD were positively correlative with the apoptosic hepatocytes. CONCLUSION: TNF-alpha induces hepatocyte apoptosis by up regulation expression of FADD protein.

Adaptor Proteins, Signal Transducing↗

Microsatellite analysis of the Queensland fruit fly Bactrocera tryoni (Diptera: Tephritidae) indicates spatial structuring: implications for population control.

The population structure of a tephritid pest species, the Queensland fruit fly Bactrocera tryoni (Froggatt), has been analysed over a five year period (1994-1998), using six microsatellites. Adult fly samples were collected to cover most regions of eastern and central Australia where the flies are regularly found. Tests for heterogeneity indicated that flies within geographically defined regions were homogeneous. The samples were allocated into five regions, including one very large region, Queensland, which encompasses that portion of the fly's range where breeding can occur year-round. With one exception, the collections from different regions were homogeneous between years, showing a fairly static distribution of the species. However, differences between regions were highly significant. The one case of a change in frequency between years indicated a gradual replacement of flies in a marginal region by flies from the main part of the range. The finding of stability in the distribution of a highly mobile insect is of interest, potentially also for other species which have expanded beyond their native range. It is argued that a contributing reason for this stability may be adaptation to different climatic regimes, and that strategies for control based on this hypothesis afford a reasonable chance of success.

Animals↗

[Change of inhabitant dietary pattern in Shandong province during 1988-1997].

Food consumption data of some bigger cities and towns of Shandong province during 1988-1997 had been collected and analyzed. The results showed that the consumption of cereals and vegetable in 1997 tended to be decreased and the consumption of meat, egg, fruit and milk in 1997 tended to be increased. The desirable dietary pattern (DDP) scores for cities and towns were 82.5, 78.7 in 1988, and 93.3, 88.4 in 1997 respectively. All these changes indicated that the dietary quality improved markedly but the consumption of bean and milk are not enough.

China↗

[Preparation and quality control of cross-linked agarose coated activated charcoal (CAAC-II) used as adsorbents for hemoperfusion in artificial liver support].

The preparation and the quality control of cross-linked agarose coated activated (CAAC-II) were described. The crosslinking reagent epichlorohydrin was removed. Investigations indicate that the trace elements such as Pb, Cd, As, Cr, Fe and heavy metals, the pH, NH4+, SO4(2-), and ultraviolet absorbance in sample have met the requirements in Chinese Pharmacopeia and some other relevant standard promulgated by the Ministry of Health. No epichlorohydrin was detected in CAAC-II.

Capsules↗

[Immune response induced by hepatitis B DNA vaccine pCI-S-IRES- ProT alpha].

OBJECTIVE: To investigate immune response in mice inoculated with DNA vaccine. METHODS: pCI-S and pCI-S-IRES- ProT alpha were constructed by gene technology, transferred into cell line NIH-3T3 mediated by lipofectamine. Mice were inoculated with these plasmids too. HBsAg and HBsAb were detected by ELISA, and transcriptions of these plasmids were detected by RT-PCR. RESULTS: pCI-S -IRES- ProT alpha, and pCI-S were effectively expressed in cultured cells and vaccinated animals. Humoral immune and specific T-cell proliferative responses were stronger in pCI-S-IRES group than pCI-S group. CONCLUSIONS: Plasmids coexpression of prothymosin alpha and hepatitis B surface antigen can improve immune responses significantly.

Animals↗

Contributions of biofilm versus suspended bacteria in an aerobic circulating-bed biofilm reactor.

This study demonstrated that, during the two-step biodegradation of toluene in an aerobic circulating-bed biofilm reactor, biofilm and suspended bacteria played critical roles. Although the suspended bacteria were less than 1% of the total amount of biomass in the system, they transformed up to 30% of the toluene into its intermediate in the bulk liquid phase. On the other hand, most of the toluene intermediate was removed inside the biofilm, where diffusion resistance reduced the toluene concentration, thereby relieving inhibition to the degradation reaction of the intermediate. The suspended bacteria are most important for rapidly biodegraded substrates, for which diffusion limitation controls the kinetics in the biofilm. They lose importance when the effects of an inhibiting substrate must be overcome.

Bacteria, Aerobic↗

The effect of fluid shear stress on ICAM-1 expression of rat brain microvascular endothelial cells.

Intercellular adhesion molecule-1 (ICAM-1) is an adherence molecule that is an important factor in many pathophysiological processes such as atherosclerosis, thrombosis and inflammation. It is secretion of endothelial cells by a variety of biochemical stimulations. But hemodynamic forces can also induce various functional changes in vascular endothelium. Some researches have proved that shear stress can modulate the expression of ICAM-1. But most of them examine the regulation of expression of ICAM-1 in human umbilical vein endothelial cells. There is no detail on the effect of shear stress (SS) on ICAM-1 expression of microvascular endothelial cells (RBMECs). In this experiment, we use cultured rat brain microvascular endothelial cells (RBMECs). By using the parallel plate flow chamber method, we give two magnitudes of lamminar shear stresses (0.2 dyn/cm2, 0.4 dyn/cm2) for different perieods of time on the slides of cells. Immunostaining method and image analysis shows a specific upregulation in ICAM-1 expression on RBMECs, which is different from endothelial cells of other species or vascular beds. Expression of ICAM-1 is increased 0.5h after the onset of SS, and reached its highest level 4h after onset of SS, then declines after that. The effect is time-dependent, not force magnitude-dependent. Endothelial cell surface expression of ICAM-1 in the supernatants of RBMECs exposed to SS was not modified excluding the possibility that RBMECs exposed to SS synthesize factors that upregulate ICAM-1. The experiment data are relevant to the current understanding of basic mechanisms that explain the signal transudation pathway occurring inside the endothelial cells under the effect of SS.

Animals↗

[Modification of the Photobacterium phosphoreum toxicity test method].

In order to reduce the variation of Photobacterium phosphoreum luminous intensity during testing, the Photobacterium phosphoreum toxicity test was modified by a correction factor, PIacetone, in this paper. The effects of magnetic stirring time, culture time, culture generation and culture temperature on the measured results were discussed. The modified method had good laboratory repeatability and the relative standard deviation (RSD) was between 2.1% and 13.1%. Reliable data could be obtained using the modified method. It would be beneficial to the development of the quantitative structure-activity relationships on multicomponent organic compounds.

Luminescent Measurements↗

[The growth hormone and insulin-like growth factors axis in liver failure patients].

OBJECTIVE: To measure circulating concentration of growth hormone (GH), insulin-like growth factors 1 (IGF-1), and it's binding protein 1 (IGFBP1) and binding protein 3 (IGFBP3) in patients with sever hepatitis, and to survey the clinical significance of GH-IGFS axis. METHODS: The study population consisted of 18 patients with liver failure caused by serious virous hepatitis, and of 20 normal volunteers. Serum concentration of GH, IGF-1, IGFBP1 and IGFBP3 was determined by ELISA. Liver biochemistric functions were measured by routine methods. RESULTS: Serum concentrations of IGF-1 and IGFBP3 were equally reduced (5.5microgram/ml +/- 6.2 microgram/ml vs 17.6 microgram/ml +/-7.0 microgram/ml, and 2.4 microgram/ml +/-1.3 microgram/ml vs 9.4 microgram/ml +/- 1.7microgram/ml, P<0.001, respectively); increased serum GH and IGFBP1 were observed (9.1ng/ml +/-12.4ng/ml vs 1.6ng/ml +/-2.4ng/ml, P<0.05; 67.9ng/ml+/-50.2ng/ml vs 45.8ng/ml+/-33.1ng/ml, P<0.01) in liver failure patients as compared with the controls. The positive relationship between IGF-1 and IGFBP3 (r=0.91, P<0.001) was observed. The closed relation between the serum IGF-1 and the prognosis of patients was also obtained. IGF-1<10 microgram/ml was in accord with 90% accurate rate of predicted death. CONCLUSIONS: The GH-IGFs axis is significantly abnormal in liver failure patients, suggesting the existence of GH resistance in these patients. Serum IGF-1 may act as a predictor for their prognosis.

Adult↗