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Biomedical subjects

D Qiu

Publications and source records attributed to D Qiu.

At least 19 recordsLinked to original sources

A comparative linkage map of oilseed rape and its use for QTL analysis of seed oil and erucic acid content.

We have developed a new DH mapping population for oilseed rape, named TNDH, using genetically and phenotypically diverse parental lines. We used the population in the construction of a high stringency genetic linkage map, consisting of 277 loci, for use in quantitative genetic analysis. A proportion of the markers had been used previously in the construction of linkage maps for Brassica species, thus permitting the alignment of maps. The map includes 68 newly developed Sequence Tagged Site (STS) markers targeted to the homologues of defined genes of A. thaliana. The use of these markers permits the alignment of our linkage map with the A. thaliana genome sequence. An additional 74 loci (31 newly developed STS markers and 43 loci defined by SSR and RFLP markers that had previously been used in published linkage maps) were added to the map. These markers increased the resolution of alignment of the newly constructed linkage map with existing Brassica linkage maps and the A. thaliana genome sequence. We conducted field trials with the TNDH population at two sites, and over 2 years, and identified reproducible QTL for seed oil content and erucic acid content. The results provide new insights into the genetic control of seed oil and erucic acid content in oilseed rape, and demonstrate the utility of the linkage map and population.

Brassica napus↗

Continuous cytomegalovirus seroconversion in a large group of healthy blood donors.

BACKGROUND AND OBJECTIVES: Transmission of cytomegalovirus (CMV) to seronegative, immunocompromised recipients can cause serious and fatal complications. Although the seroprevalence of CMV is high, the risk of primary CMV infection among healthy blood donors has not yet been analysed in a large population. MATERIALS AND METHODS: We developed an algorithm to determine the rate of CMV seroconversion in an overall cohort of 24,260 subjects who donated 176,474 blood units during an 11-year observation period. RESULTS: We detected CMV seroconversion in all relevant age groups (18-60 years) with an overall seroconversion rate of 0.55% per year. Both CMV seroconversion and seroprevalence occurred more frequently in female donors (P = 0.02 and P < 0.001, respectively). We identified 30-35-year-old blood donors as the group with the highest rate of CMV seroconversion per year (1.33% vs. 0.46%; P < 0.0001). CONCLUSIONS: We conclude that the risk of primary CMV infection is a continuous lifelong event and correlates with age and female gender.

Adolescent↗

Estimating the distribution of worm burden and egg excretion of Schistosoma japonicum by risk group in Sichuan Province, China.

During autumn 2000 an extensive cross-sectional survey of the prevalence of Schistosomiasis japonicum was conducted among about 4000 villagers within 20 villages in the Anning River Valley located in the southwestern Sichuan Province. Two procedures were used to assess infection status, the Kato-Katz thick smear procedure and a miracidia hatch test. Whereas the Kato-Katz procedure provides information on both prevalence and intensity, the hatch test provides only prevalence data, albeit on a much larger volume of stool. In addition, we performed Kato-Katz smears for 15 consecutive samples on a subset of 15 individuals. The proportion of both hatch-test and Kato-Katz positive individuals in the larger cross-sectional survey was 25%. The goal of the study was to estimate both the egg and worm distributions among risk groups using both the hatch and Kato-Katz tests from the cross-sectional data and the repeated Kato-Katz smears from the longitudinal data sets. As a prelude to parameter estimation, individuals were classified into risk groups by natural village and occupation; the proportion of Kato-Katz positive subjects among the risk groups varied from 10% to 60%. We used the statistical model of de Vlas et al. (1992) and Bayesian techniques to derive both estimates of and inference about the worm and egg distribution parameters. The parameter estimates imply (1) similar eggs per gram stool (e.p.g.) per worm pair compared with earlier estimates, (2) a range of worm burdens among the risk groups and (3) estimates of risk heterogeneity within groups is sensitive to prior information on the within-person variability in egg excretion.

Age Factors↗

Dynamics of single and multiple bubbles and associated heat transfer in nucleate boiling under low gravity conditions.

Experimental studies and numerical simulation of growth and lift-off processes of single bubbles formed on designed nucleation sites have been conducted under low-gravity conditions. Merging of multiple bubbles and lift-off processes during boiling of water in the parabola flights of KC-135 aircraft were also experimentally studied. The heating area of the flat heater surface was discretized and equipped with a number of small heating elements that were separately powered in the temperature-control mode. As such, the wall superheat remained nearly constant during the growth and departure of the bubbles, whereas the local heat flux varied during the boiling process. From numerical calculation it is found that peak of heat flux occurs locally at the contact line of bubble and heater surface. Dry conditions exist inside the bubble base area, which is characterized through a zero heat flux region in the numerical calculation and a lower heat flux period in the experimental results. During the merger of multiple bubbles, dry-out continues. In both the numerical calculations and experimental results, the bubble lift-off is associated with an apparent increase in heat flux. Wall heat flux variation with time and spatial distribution during the growth of a single bubble from numerical simulations are compared with experimental data.

Journal Article↗

Comparison of different methods of total RNA extraction for viral detection in sputum.

Examination of sputum specimens can be used for monitoring airway inflammation by means of immunological and molecular techniques. RNA extraction is essential for measurement of cytokine gene expression and for detection of many viral pathogens in sputum. In this study, three RNA extraction methods used commonly (acid guanidinium thiocyanate-phenol-chloroform extraction, Trizol and RNeasy Mini kit) were compared on the sputum of 14 patients who had chronic obstructive pulmonary disease. The effect of dithiothreitol pre-treatment on sputum RNA extraction was also investigated. The yield and purity of total RNA were determined by spectrophotometry. Reverse transcription-polymerase chain reaction (RT-PCR) results of the house keeping gene (glyceraldehyde-3-phosphate dehydrogenase) and RNA sequences specific to common respiratory viruses were compared. The results showed that (1) total RNA extracted with Trizol had highest yield and purity among the three RNA extraction methods; (2) there was no significant difference among the three RNA extraction methods on the house keeping gene and viral detection by RT-PCR; (3) dithiothreitol pre-treatment did not improve either the purity of total RNA, or RT-PCR signal. Moreover, dithiothreitol treatment reduced significantly the yield of total RNA. The results of the study indicate that the Trizol method appears to be superior for total RNA extraction from sputum, and dithiothreitol pre-treatment does not increase the efficiency of RNA extraction and RNA detection in sputum specimens.

Dithiothreitol↗

Genome sequence and comparative analysis of the solvent-producing bacterium Clostridium acetobutylicum.

The genome sequence of the solvent-producing bacterium Clostridium acetobutylicum ATCC 824 has been determined by the shotgun approach. The genome consists of a 3.94-Mb chromosome and a 192-kb megaplasmid that contains the majority of genes responsible for solvent production. Comparison of C. acetobutylicum to Bacillus subtilis reveals significant local conservation of gene order, which has not been seen in comparisons of other genomes with similar, or, in some cases closer, phylogenetic proximity. This conservation allows the prediction of many previously undetected operons in both bacteria. However, the C. acetobutylicum genome also contains a significant number of predicted operons that are shared with distantly related bacteria and archaea but not with B. subtilis. Phylogenetic analysis is compatible with the dissemination of such operons by horizontal transfer. The enzymes of the solventogenesis pathway and of the cellulosome of C. acetobutylicum comprise a new set of metabolic capacities not previously represented in the collection of complete genomes. These enzymes show a complex pattern of evolutionary affinities, emphasizing the role of lateral gene exchange in the evolution of the unique metabolic profile of the bacterium. Many of the sporulation genes identified in B. subtilis are missing in C. acetobutylicum, which suggests major differences in the sporulation process. Thus, comparative analysis reveals both significant conservation of the genome organization and pronounced differences in many systems that reflect unique adaptive strategies of the two gram-positive bacteria.

Amino Acid Sequence↗

[Expression of neural cell adhesion molecule in murine livers with experimental autoimmune hepatitis].

OBJECTIVE: To investigate the expression of neural cell adhesion molecule (NCAM) in murine livers with experimental autoimmune hepatitis (EAH) and its relationship with histological grades. METHODS: To induce the EAH model, the syngeneic S-100 antigen emulsified in complete Freud's adjuvant was injected intraperitoneally to C57Bl/6 at day 1 and day 7. The mice were sacrificed at day 7, 14, and 21 after the first immunization. The expression of NCAM was observed using histoimmunochemistry and RT-PCR. RESULTS: The expression of NCAM increased gradually with aggravation of lymphocyte infiltration and hepatocyte injury. The administration of prednisolone decreased the histological grade, and inhibited the expression of NCAM. There was a correlation between the expression of NCAM and histological grade (r=0.71, P<0.01). CONCLUSIONS: NCAM provides a traffic signal for infiltration of lymphocytes in the liver, hence induces the injury of hepatocytes.

Animals↗

[Time for extubation and sequential noninvasive mechanical ventilation in COPD patients with exacerbated respiratory failure who received invasive ventilation].

OBJECTIVE: To discuss the optimal time for extubation and sequential non-invasive mechanical ventilation in COPD patients with exacerbated respiratory failure who received invasive ventilation. METHODS: 24 patients received invasive ventilation for 3 days were randomly assigned to receive noninvasive ventilation (study group) or to continue the weaning process with invasive ventilation (control group). The incidence of ventilator-associated pneumonia (VAP), the incidence of death, the duration of ventilation, the hospitalization were analyzed in two groups. RESULTS: Between study group and control group, the incidence of VAP was 0/12 vs 7/12, P = 0.027; the incidence of death was 0/12 vs 3/12, P = 0.217; the continued duration of ventilation after invasive ventilation for 3 days was (7 +/- 5) days vs (15 +/- 12) days, P < 0.05; the hospitalization was (16 +/- 6) days vs (25 +/- 12) days, P < 0.05, respectively. CONCLUSIONS: In COPD patients with exacerbated respiratory failure who received invasive ventilation, invasive ventilation for 3 days followed by noninvasive ventilation may decrease the incidence of VAP, shorten the duration of ventilation and hospitalization.

Aged↗

[Polycystic ovaraian syndrome and hyperinsulinemia].

OBJECTIVE: To identify the relationship between polycystic ovarian syndrome and hyperinsulinemia. METHODS: Age, body height, body weight, serum estradiol, progesterone, testosterone, follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), triiodothyronine (T3), tetraiodothyronine (T4), insulin, oral glucose tolerance test (OGTT), and cortisol were measured in 28 patients with polycystic ovarian syndrome (PCOS). RESULTS: Sxiteen patients (57.1% of total) had hyperinsulinemia. Among them one patient was found diabetic. No difference was noted in age, height, body weight, body mass index, serum estradiol, progesterone, testosterone, FSH, LH, PRL, T3, T4, cortisol, and glucose of OGTT between hyperinsulinemia and non-hyperinsulinemia patients. However the LH/FSH ratio (2.4 +/- 1.5) in hyperinsulinemia patients was significantly higher than 1.2 +/- 0.6 in non-hyperinsulinemia patients (P = 0.0125). In addition, the results showed that the fasting serum insulin, insulin after OGTT, the insulin/glucose ratio at 30, 120 minutes, 180 minutes, the area under curve of and the insulin, insulin resistant index (IRI) were higher in hyperinsulinemia pateints than in non-hyperinsulinemia patients respectively. Furthermore, the fasting insulin level, insulin/glucose ratio were found to be negatively related to insulin sensitivity index and positively related with beta-cell functioning index. CONCLUSION: Hyperinsulinemia and insulin resistance accompany with some of the PCOS patients in China. Further investigations will be necessary to clarify the relevant mechanisms.

Adult↗

Agrin controls synaptic differentiation in hippocampal neurons.

Agrin controls the formation of the neuromuscular junction. Whether it regulates the differentiation of other types of synapses remains unclear. Therefore, we have studied the role of agrin in cultured hippocampal neurons. Synaptogenesis was severely compromised when agrin expression or function was suppressed by antisense oligonucleotides and specific antibodies. The effects of antisense oligonucleotides were found to be highly specific because they were reversed by adding recombinant agrin and could not be detected in cultures from agrin-deficient animals. Interestingly, the few synapses formed in reduced agrin conditions displayed diminished vesicular turnover, despite a normal appearance at the EM level. Thus, our results demonstrate the necessity of agrin for synaptogenesis in hippocampal neurons.

Agrin↗

Regulation of agrin expression in hippocampal neurons by cell contact and electrical activity.

Most synapses contain high concentrations of neurotransmitter receptors in the postsynaptic plasma membrane. Agrin (Ag) is an extracellular matrix protein necessary for the localization of acetylcholine receptors at the neuromuscular junction and for the differentiation of synapses in hippocampal neurons in vitro. The temporal pattern of agrin expression during the development of the central nervous system (CNS) is consistent with the notion that agrin expression is regulated during synaptogenesis. To identify the processes underlying this regulation, we have analyzed levels and alternative splicing of agrin mRNA in primary hippocampal neurons. Our results indicate that in the initial phases of synapse formation, contact-mediated processes and action potential-dependent neurotransmission regulate agrin mRNA expression, while secreted factors from glial cells, but not from hippocampal neurons, influence the alternative splicing of agrin mRNA. Previous studies have shown that specific agrin isoforms are able to induce the activation of a transcription factor and that secreted agrin associates with cellular surfaces. Therefore, we have tested whether agrin isoforms contribute to the contact-mediated induction of agrin expression in hippocampal neurons. None of the agrin isoforms tested in this study revealed this activity. Finally, we show that the role of evoked neural transmission in controlling agrin transcription changes during differentiation in vitro.

Action Potentials↗

Purkinje cell fate in staggerer mutants: agenesis versus cell death.

Staggerer (sg/sg) is an autosomal recessive mutation in an orphan nuclear hormone receptor gene, RORalpha, that causes a cell-autonomous, lineage-specific block in the development of the Purkinje cell. Purkinje cell number is reduced by about 75-90% in adult mutants, and many of the surviving cells are small and ectopically positioned. To determine whether Purkinje cell numbers are reduced owing to either agenesis or cell death, cohorts of Purkinje cells were labeled with the birth-date marker bromodeoxyuridine (BrdU) at embryonic day (E) 10.5 or E11.5. The total number of BrdU-labeled profiles was then compared between cerebella from wild-type mice, heterozygous staggerer, and staggerer mutants at E17.5 and postnatal day (P)5. There was no significant difference between sg/sg mutants and +/sg or +/+ controls in the number of BrdU-labeled profiles or in cerebellar volumes in the E17 embryos. By P5, however, cerebellar volume was significantly reduced in the sg/sg mutants compared to controls (p <.005) and the number of BrdU-labeled profiles was reduced by 33% following E11.5 BrdU injections (p <.02). The results suggest that Purkinje cell genesis is not affected by the staggerer mutation and that Purkinje cell loss begins some time after E17. RORalpha is highly expressed in Purkinje cells by E14, so the delay between initial RORalpha expression and sg/sg Purkinje cell loss suggests that the staggerer mutation does not directly cause Purkinje cell death.

Aging↗

Characterisation of membrane phospholipids and glycolipids from a halophilic archaebacterium by high-performance liquid chromatography/electrospray mass spectrometry.

Combined high-performance liquid chromatography and electrospray mass spectrometry (LC/ES-MS) has been used for direct characterisation of the polar membrane lipids in total lipid extracts from Halobacterium salinarium, a species of halophilic archaebacterium. The principle phospholipids found were the diphytanyl archaeol phosphatidylglycerol and diphytanyl archaeol phosphatidylglycerolphosphate methyl ester. The application of LC/ES-MS revealed the additional presence of diphytanyl archaeol phosphatidylglycerol sulphate The extracts also contained an archaeol glycolipid, initially detected in preliminary offline ES-MS studies, which was further characterised by LC/ES-MS and by product ion tandem mass spectrometry (MS/MS) as a sulphate ester of diglycosyl-2,3-di-O-phytanyl-sn-glycerol. Whilst archaeol phospho- and glycolipids containing a (C(20)-C(20))-isopranyl glycerol ether core predominated, LC/ES-MS of the extracts from Halobacterium salinarium indicated the presence of an analogue containing one double bond in its isoprenyl ether core as a minor component of the phosphatidylglycerolphosphate methyl ester fraction, providing a further example of the previously recognised existence of isoprenologues of diphytanyl archaeols which occur as minor components of archaebacterial membrane lipids. The value of these techniques in compositional analysis of archaebacterial lipid extracts is discussed.

Archaea↗

Glycosylations versus conformational preferences of cancer associated mucin core.

Synthetic oligosaccharide vaccines based on core STn (sialyl alpha2-6 GalNAc) carbohydrate epitopes are being evaluated by a number of biopharmaceutical firms as potential immunotherapeutics in the treatment of mucin-expressing adenocarcinomas. The STn carbohydrate epitopes exist as discontinuous clusters, O-linked to proximal serine and threonine residues within the mucin sequence. In an effort to probe the structure and dynamics of STn carbohydrate clusters as they may exist on the cancer-associated mucin, we have used NMR spectroscopy and MD simulations to study the effect of O-glycosylation of adjacent serine residues in a repeating (Ser)n sequence. Three model peptides/glyco-peptides were studied: a serine trimer containing no carbohydrate groups ((Ser)3 trimer); a serine trimer containing three Tn (GalNAc) carbohydrates alpha-linked to the hydroxyls of adjacent serine sidechains ((Ser.Tn)3 trimer); and a serine trimer containing three STn carbohydrates alpha-linked to the hydroxyls of adjacent serine sidechains ((Ser.STn)3 trimer). Our results demonstrate that clustering of carbohydrates shifts the conformational equilibrium of the underlying peptide backbone into a more extended and rigid state, an arrangement that could function to optimally present the clustered carbohydrate antigen to the immune system. Steric effects appear to drive these changes since an increase in the size of the attached carbohydrate (STn versus Tn) is accompanied by a stronger shift in the equilibrium toward the extended state. In addition, NMR evidence points to the formation of hydrogen bonds between the peptide backbone NH protons and the proximal GalNAc groups in the (Ser.Tn)3 and (Ser.STn)3 trimers. The putative peptide-sugar hydrogen bonds may also play a role in influencing the conformation of the underlying peptide backbone, as well as the orientation of the O-linked carbohydrate. The significance of these results will be discussed within the framework of developing clustered STn-based vaccines, capable of targeting the clustered STn epitopes on the cancer-associated mucin.

Adenocarcinoma↗

Anti-inflammatory effects of triptolide in human bronchial epithelial cells.

Triptolide (PG490, 97% pure) is a diterpenoid triepoxide with potent anti-inflammatory and immunosuppressive effects in transformed human bronchial epithelial cells and T cells (Qiu D, Zhao G, Aoki Y, Shi L, Uyei A, Nazarian S, Ng JC-H, and Kao PN. J Biol Chem 274: 13443-13450, 1999). Triptolide, with an IC(50) of approximately 20-50 ng/ml, inhibits normal and transformed human bronchial epithelial cell expression of interleukin (IL)-6 and IL-8 stimulated by phorbol 12-myristate 13-acetate (PMA), tumor necrosis factor-alpha, or IL-1 beta. Nuclear runoff and luciferase reporter gene assays demonstrate that triptolide inhibits IL-8 transcription. Triptolide also inhibits the transcriptional activation, but not the DNA binding, of nuclear factor-kappa B. A cDNA array and clustering algorithm analysis reveals that triptolide inhibits expression of the PMA-induced genes tumor necrosis factor-alpha, IL-8, macrophage inflammatory protein-2 alpha, intercellular adhesion molecule-1, integrin beta(6), vascular endothelial growth factor, granulocyte-macrophage colony-stimulating factor, GATA-3, fra-1, and NF45. Triptolide also inhibits constitutively expressed cell cycle regulators and survival genes cyclins D1, B1, and A1, cdc-25, bcl-x, and c-jun. Thus anti-inflammatory, antiproliferative, and proapoptotic properties of triptolide are associated with inhibition of nuclear factor-kappa B signaling and inhibition of genes known to regulate cell cycle progression and survival.

Algorithms↗

The human bone marrow response to acute air pollution caused by forest fires.

Atmospheric pollution increases cardiopulmonary morbidity and mortality by unexplained mechanisms. Phagocytosis of fine particles (PM(10)) by rabbit alveolar macrophages elevates white blood cells (WBC) by releasing precursors from the bone marrow and this could contribute to the pathogenesis of cardiopulmonary disease. The present study examined the association between acute air pollution caused by biomass burning and peripheral WBC counts in humans. Serial measurements of the WBC count made during the 1997 Southeast Asian Smoke-haze (Sep 29, Oct 27) were compared with a period after the haze cleared (Nov 21, Dec 5) using peripheral blood PMN band cells to monitor marrow release. The results showed that indices of atmospheric pollution were significantly associated with elevated band neutrophil counts expressed as a percentage of total polymorphonuclear leukocytes (PMN), with maximal association on zero and 1 lag day for PM(10) and 3, and 4 lag days for SO(2) (p value < 0.000). We conclude that atmospheric pollution caused by biomass burning is associated with elevated circulating band cell counts in humans because of the increased release of PMN precursors from the marrow. We speculate that this response contributes to the pathogenesis of the cardiorespiratory morbidity associated with acute air pollution.

Adult↗

Effects of lipid on low-density and high-density lipoprotein receptors in hepatic stellate cell from rat liver.

OBJECTIVE: To study the effects of lipid (triglyceride and very low-density lipoprotein) on low-density lipoprotein (LDL) and high-density lipoprotein (HDL) receptors in the hepatic stellate cell (HSC) from the rat liver. METHODS: HSC were isolated and cultured from the liver of Wistar rats by in situ perfusion with pronase and collagenase and density gradient centrifugation with Nycodenz. Radioligand conjugation assay with (125)I-LDL and (125)I-HDL(3) was detected for the effects of lipid on LDL and HDL receptor of HSC. RESULTS: LDL and HDL receptors were found on the membrane of the rat HSC. The lipid might increase the binding of LDL to LDL receptor, but decrease the binding of HDL(3) to HDL receptor. CONCLUSION: LDL and HDL receptors on the HSC membrane may have an important role in the metabolism of lipoprotein and the regulation of cholesterol. These results provided the basis of theory and experimentation for the genesis of fatty liver and liver fibrosis.

Animals↗

Transfection and expression of HCV-NS5B gene in Huh-7 cells.

OBJECTIVE: To study the mechanism of hepatitis C virus (HCV) replication and to test gene therapy for hepatitis C, a human liver cell line expressed HCV RNA polymerase has been established. METHODS: NS5B gene has been transfected into Huh-7 cells by lipofectamine. The results of transfection were confirmed by PCR and Southern blot analysis, and the level of the non-structural protein 5B (NS5B) in Huh-7 cells was detected by Western blot analysis. RESULTS: There were NS5B gene fragments and the expression of NS5B protein in Huh-7c cells transfected with pTeT-NS5B or pcDNA-NS5B plasmid. CONCLUSIONS: We have established a HCV RNA polymerase expression system in Huh-7 cells which can be further used to analyze the mechanism of HCV replication and provide a cell model for gene therapy in vitro.

Carcinoma, Hepatocellular↗