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Biomedical subjects

Q Y Chen

Publications and source records attributed to Q Y Chen.

At least 19 recordsLinked to original sources

Using large eddy simulation to study particle motions in a room.

UNLABELLED: As people spend most of their time in an indoor environment, it is important to predict indoor pollutant level in order to assess health risks. As particles are an important pollutant indoors, it is of great interest to study the airflow pattern and particle dispersion in buildings. This study uses large eddy simulation (LES) to predict three-dimensional and transient turbulent flows and a Lagrangian model to compute particle trajectories in a room. The motion of three different types of solid particles in a decaying homogeneous isotropic turbulent airflow is calculated. By comparing the computed results with the experimental data from the literature, the computational method used in this investigation is found to be successful in predicting the airflow and particle trajectories in terms of the second-order statistics, such as the mean-square displacement and turbulent intensity. This Lagrangian model is then applied to the study of particles' dispersion in a ventilated cavity with a simplified geometry for two ventilation scenarios. It is shown that light particles follow the airflow in the room and many particles are exhausted, while heavier particles deposit to the floor or/and are exhausted. PRACTICAL IMPLICATIONS: The results of this paper can be used to study dispersion of infectious diseases in enclosed spaces in which virus or bacteria are often attached to particles and transported to different rooms in a building through ventilation systems. In most of studies, the virus or bacteria have been considered to be gaseous phase so there is no slip between virus/bacteria and air. The results in this paper show that heavier particles are submitted to gravity and are sensitive to the ventilation strategy.

Air Movements↗

Inhibitory potential of herbal medicines on human cytochrome P450-mediated oxidation: properties of umbelliferous or citrus crude drugs and their relative prescriptions.

To investigate the possible drug interaction with herbal medicine, hot water decoctions or 40% ethanol infusions of several Umbelliferous or Citrus crude drugs and their prescriptions were examined in vitro for their abilities to inhibit human cytochrome P450 3A (CYP3A). Addition of each decoction or infusion from Baizhi (Angelica dahurica and varieties), Qianghuo (Notopterygium incisum or N. forbesii), Duhuo (Angelica biserrata), Fangfeng (Saposhnikovia divaricata), Danggui (Angelicasinensis), Zhishi or Zhiqiao (Citrus aurantium) resulted in various degrees of human CYP3A inhibition as determined by microsomal testosterone 6beta-hydroxylation. The inhibitory potency was consistent with the abundance of the hydrophobic components for each sample. Experiments on the infusion of a Japanese Baizhi (BZ1) showed the major role of furanocoumarins on human CYP3A inhibition. Some of the crude drugs and a related prescription showed increased inhibition after the preincubation, suggesting the involvement of a mechanism-based inhibition. Some formulated prescriptions, however, showed intense inhibition with their hydrophobic fractions rather than with their hydrophobic fractions, suggesting that components other than furanocoumarins in herbal prescriptions may also cause CYP3A inhibition. These results indicate the necessity of intensive investigations on the possible drug interaction with traditional medicines.

Aryl Hydrocarbon Hydroxylases↗

A novel method for the determination of total protein in human serum by near infrared fluorescence recovery.

A novel fluorometric method has been developed for the determination of total protein in human serum with a new near-IR reagent as a fluorescence probe, based on the fluorescence recovery of the cyanine-CTAB system in the presence of protein. Maximum fluorescence is produced with maximum excitation and emission wave-lengths at 765 and 812 nm, respectively. Under optimal conditions, the calibration graphs are linear over the range 0.4-12.0 microg/mL for protein. The detection limit is 70 ng/mL, and the relative standard deviation of six replicate measurements is 1.14% for 6.0 microg/mL protein. The results are satisfactory.

Animals↗

Application of magdala red as a fluorescence probe in the determination of nucleic acids.

A fluorescence quenching method was developed for the rapid determination of DNA and RNA using magdala red as fluorescence probe. In weakly acidic medium, the fluorescence of magdala red (lambdaex/lambdaem = 540/555 nm) can be largely quenched by DNA or RNA. The calibration graphs are linear over the range 0.01-1.2 microg/mL for both calf thymus DNA (CT DNA) and salmon DNA (SM DNA), and 0.015-1.0 microg/mL for yeast RNA, respectively. The corresponding detection limits are 6.0 ng/mL for CT DNA, 7.0 ng/mL for SM DNA and 15.0 ng/mL for yeast RNA, respectively. CT DNA could be determined in the presence of 20% (w/w) yeast RNA, and the relative standard deviation of six replicate measurements is 3.18% for 400 ng/mL of CT DNA. Interference from coexisting substances in the determination of DNA was also examined. Real samples were determined with satisfactory results.

Adenine↗

Determination of proteins at nanogram levels by their quenching effect on large particle scattering of colloidal silver chloride.

A novel quantitative method for the determination of proteins in aqueous solutions has been based on the quenching of the resonance scattering light of colloidal silver chloride in the presence of proteins. The detection limits for eight kinds of proteins (BSA, HSA, egg albumin, human gamma-IgG,alpha-chymotrypsin, E. Coli. alpsase, myoglobin, alpha-casein) were at about 8 ng/mL; the linear ranges of the calibration curves were 10-400 ng/mL under optimal conditions,except for human gamma-IgG (20-400 ng/mL), myoglobin (10-300 ng/mL), and alpha-casein (10-300 ng/mL). Three wavelengths (398 nm, 475 nm, 499 nm) were all suitable for the determination and any acidity from pH 3.0 to pH 9.0 could be chosen. A few non-protein substances at high concentration levels interfered with this method, but this problem could simply be overcome by diluting the samples before the assay. Mechanism studies showed that the quenching effect of proteins on the scattering light of colloidal silver chloride was mainly due to the coagulation of AgCl particles retarded by protein. The method was employed for the determination of total protein in human serum with satisfactory results.

Animals↗

A novel mimetic enzymatic fluorescence immunoassay for hepatitis B surface antigen by using a thermal phase separating polymer.

Iron tetrasulfonatophthalocyanine (FeTSPc), a peroxidase mimic, was used as a labeling reagent and poly(N-isopropylacrylamide) (PNIP) as the separation support of the immune complex for the mimetic-enzymatic immunoassay of hepatitis B surface antigen (HBsAg). PNIP was precipitated from aqueous solution when the ambient temperature was higher than its lower critical solution temperature of 31 degrees C. In a sandwich immunoassay, the antigen (HBsAg) first reacted with mouse anti-human HBsAg antibody immobilized on PNIP (PNIP-antibody) and then further reacted with FeTSPc-labeled mouse anti-HBsAg antibody (antibody-FeTSPc) at room temperature in a homogeneous format. After changing the temperature to separate the PNIP-antibody-HBsAg-antibody-FeTSPc conjugate moiety, it was re-dissolved and determined by coupling with the fluorogenic reaction of hydrogen peroxide and p-hydroxyphenylpropionic acid. The sensitivity of this method (3 ng mL-1) was close to that of the traditional ELISA using the same reactants. However, the assay was much faster (the assay time decreased from 100-120 to 45 min). This method was applied to determine HBsAg in human serum with satisfactory results.

Hepatitis B Surface Antigens↗

The human leukocyte antigen HLA DRB3*020/DQA1*0501 haplotype is associated with Graves' disease in African Americans.

Information on genetic susceptibility to Graves' disease in African Americans is limited. We studied DRB1, DQB1, DRB3 subtypes, DQA1*0501, DQA1*0201, and CTLA-4 polymorphisms in 49 African American patients with adult onset Graves' disease and 47 racially-matched controls using PCR-based sequence-specific priming methods. There were no significant differences in DRB1 or DQB1 allelic frequencies or CTLA-4 polymorphisms between patients and controls. However, we found that the frequency of DRB3 was significantly increased in the patients (75.5% vs. 57.4%, P = 0.006, X2 = 3.52), especially for the DRB3*0202 subtype (53.1% vs. 23.4, P = 0.003, X2 = 8.91). In this one respect, the finding was in concordance with our previous observations in Caucasian patients with adult-onset Graves' disease. In addition, whereas the frequency of DQA1*0501 was increased (P = 0.018, X2 = 5.63) in our patients, the haplotype of DRB3/DQA1*0501, or DRB3*0202/DQA1*0501 was found to be more strongly associated (P = 0.008, X2 = 7.0; P = 0.0008, X2 = 11.34, respectively). These data suggest that DRB3*0202, particularly when found with DQA1*0501 in a haplotype is a susceptible gene(s) for Graves' disease in adult African Americans. Considering these data with those in Caucasian patients, our results would suggest that the primary Graves susceptible locus is likely DRB3 and not DRB1.

Abatacept↗

[Immunization of mice with plasmid DNA against malaria and regulation of antigen expression by tetracycline-controlled promoter].

Sequence of MSP1-31 of Plasmodium falciparum was constructed into eukaryotic expression vector pTRE, which could be repressed by tetracycline (Tc) and resulted in recombinant plasmid pTRE-31. The plasmid was injected into the quadriceps muscle of BALB/c mice with Tc responsive plasmid pTet-off to measure specific antibodies. The MSP1-31 prokaryotic expressed protein was used as antigen in ELISA. Results showed that mice orally administered by Tc had a seroconversion rate of 7.1% (1/14) 4 weeks after injection, whereas the control mice had a seroconversion rate of 100% and the titers of antibody were raised continusly within 12 weeks. The study suggested that the recombinant plasmids pTRE-31/pTet-off could efficiently induce humoral response against MSP1-31 of malaria. Moreover this immune response was controlled by Tc and was reversible after withdrawal of Tc dilivery. The induction of antibody by removing Tc at the fourth week after injection indicated that DNA vaccine could remain in mice and capable of expressing antigen for at least 4 weeks.

Animals↗

Anti-idiotypic antibodies to anti-PDC-E2 in primary biliary cirrhosis and normal subjects.

Anti-idiotypic antibodies may regulate the immune system and influence pathogenic autoimmunity. We investigated idiotype-anti-idiotype interactions in sera of patients with primary biliary cirrhosis (PBC), normal subjects and animals immunized with pyruvate dehydrogenase complex (PDC) or its derivatives. IgG autoantibody to the E2 subunit of PDC (PDC-E2) was derived by affinity-purification from sera of 12 patients with PBC, and F(ab)2 was prepared (anti-PDC-E2-F[ab]2). This was used as a reactant by enzyme-linked immunosorbent assay (ELISA) with sera from patients with PBC, normal subjects, or immunized animals. Results were that IgG antibody to anti-PDC-E2-F(ab)2 was detectable at low concentration in 12 PBC sera (mean optical density [OD] +/- SD: 1.02 +/- 0.26), and also in 19 normal sera (mean OD +/- SD: 0.97 +/- 0. 35) using a serum dilution of 1:20; background OD was 0.09 to 0.10, whereas antisera from animals immunized with PDC or PDC-E2 were nonreactive. There was a significant inverse correlation (r = -.59, P =.04) between the levels of anti-PDC-E2 in PBC sera (but not normal sera), and anti-idiotypic antibody reactive with anti-PDC-E2-F(ab)2. Anti-idiotypic antibody existed as a complex with anti-PDC-E2, because the removal of anti-PDC-E2 from serum resulted in decreased reactivity to anti-PDC-E2-F(ab)2. Reactivity between PDC-E2 and anti-PDC-E2 from PBC serum was not inhibited by normal sera, indicating that anti-idiotypic antibody from normal sera with anti-PDC-E2 reacts with the framework of F(ab) rather than the paratope. The conclusions are that PBC and normal sera contain IgG class anti-idiotypic antibodies to anti-PDC-E2, the characteristic autoantibody in PBC. Anti-PDC-E2 in immunized animals does not contain an idiotype cross-reactive with human anti-PDC-E2. Anti-idiotypic antibody in PBC is complexed with anti-PDC-E2 and in part accounts for immune complexes demonstrable in PBC. Anti-idiotypic antibody in PBC may regulate levels of anti-PDC-E2.

Adult↗

Interaction of a novel red-region fluorescent probe, Nile blue, with DNA and its application to nucleic acids assay.

A novel fluorimetric method was developed for the rapid determination of DNA and RNA based on their quenching effect on the cationic red-region fluorescent dye Nile Blue (NB). In the investigation of the interaction of NB with DNA by steady-state polarization measurements, thermal denaturing study, determination of absorption and fluorescence characteristics, salt effect study and electrophoresis experiments, the results supported the suggestion that NB served as an intercalator to the stack base pairs of nucleic acids. Further evidence showed that the quenching could be ascribed to the static quenching mode. A binding constant of about 10(6) M-1 and a binding site size of about three base pairs were obtained by spectral methods. Under optimum conditions, the calibration curves for the determination of calf thymus DNA (CT DNA) and yeast RNA were linear over the ranges 3.0 ng mL-1-2.0 micrograms mL-1 and 27 ng mL-1-10 micrograms mL-1, respectively. The detection limits were 3.0 ng mL-1 for CT DNA and 27 ng mL-1 for RNA. The relative standard deviation (n = 6) was within 2.1% in the middle of the linear range. Interferences from some interesting co-existing substances in the determination of DNA were also examined.

Animals↗

HLA-DRB1*08, DRB1*03/DRB3*0101, and DRB3*0202 are susceptibility genes for Graves' disease in North American Caucasians, whereas DRB1*07 is protective.

Graves' disease is known to be HLA-D associated; however, the primary loci involved remain unclear. We examined HLA genotypes of DRB1 and DQB1 plus DRB3 subtypes using PCR-based sequence-specific priming in two groups of North American (Gainesville, FL; and Toronto, Canada) Caucasian patients with Graves' disease. We stratified patients into those with either early age at onset (<20 yr; 13.1 +/- 4.8 yr; n = 30) and later age at onset of disease (38.8 +/- 9.7 yr; n = 62) and compared the results to 192 normal controls. As expected, we found that DRB1*03 was associated with Graves' disease, but at a higher odds ratios for early-onset than later-onset patients (3.7 vs. 2.2). The frequency of DRB1*08 was also increased in both groups of patients, but significantly so only in patients with early-onset Graves' (P = 0.001; chi2 = 10.8). DRB3 was highly associated with Graves' in both groups of patients (P = 0.009; chi2 = 6.83 and P = 0.0015; chi2 = 10.1, respectively); however, the subtypes of DRB3 revealed differential susceptibilities. Whereas the frequencies of both DRB3*0101 and DRB3*0202 were increased over the entire cohort, that of DRB3*0301 was not. Significant P values were found for DRB3*0101 in patients with early-onset and for DRB3*0202 in patients with later onset of Graves' disease. When the haplotypes of DRB1*03-DRB3 of all subtypes were removed for analysis (all DRB1*03 positive also had DRB3*0101), the frequency of DRB3*0202 remained significantly higher in the patients with later onset of Graves' disease than in controls (P = 0.0043; chi2 = 8.13), but DRB3 was no longer positively associated with the early-onset group. In addition, we found that DRB1*07 was negatively associated with both groups of patients (P = 0.024; chi2 = 5.10 and P = 0.0085; chi2 = 6.93). These data suggest that DRB3*0202 is more likely to be the primary susceptible locus than DRB1*03 for patients with later onset of Graves' disease.

Adult↗

Natural and disease associated autoantibodies to the autoantigen, dihydrolipoamide acetyltransferase, recognise different epitopes.

Naturally occurring autoantibodies are ubiquitous and may serve physiological functions. We examined the relationship of natural and disease-associated autoantibodies in the context of autoantibodies to dihydrolipoamide acetyltransferase, the 74 kDa E2 sub-unit of the mitochondrial pyruvate dehydrogenase complex (PDC-E2), characteristic of primary biliary cirrhosis (PBC). We tested for natural autoantibodies to PDC-E2 in normal sera, and compared epitopes recognised by natural and disease-associated autoantibodies. Methods included affinity purification of anti-PDC-E2 from normal and PBC sera, ELISA and immunoblotting, capacity of antibodies to inhibit the enzyme function of the pyruvate dehydrogenase complex (PDC), use of F(ab)2 fragments of anti-PDC-E2 in inhibition assays, and testing affinity purified anti-PDC-E2 on peptide fragments of PDC-E2. We found that natural auto-antibodies to PDC-E2 of IgG class were demonstrable in all healthy human sera (10/10). However, their reactivity differed from that of disease-associated autoantibodies, in that anti-PDC-E2 from normal serum failed to inhibit the catalytic activity of PDC; and F(ab)2 fragments from PBC sera potently blocked the binding of anti-PDC-E2 from PBC sera to PDC-E2, but not the binding of natural anti-PDC-E2 to PDC-E2. Immunoblotting on fragments of PDC-E2 using affinity-purified preparations from PBC sera and normal sera failed to provide evidence for gross differences in epitope reactivity. We conclude that normal human sera contain natural IgG autoantibodies to the immunodominant inner lipoyl domain of PDC-E2, as seen characteristically in PBC. However, there is evidence for differences in fine epitope recognition.

Acetyltransferases↗

The gene responsible for autoimmune polyglandular syndrome type 1 maps to chromosome 21q22.3 in US patients.

Autoimmune polyglandular syndrome type 1 [APS-1] comprises multiple organ-specific autoimmunities such as acquired hypoparathyroidism and autoimmune Addison's disease, and a predisposition to certain infections such as chronic mucocutaneous candidiasis. An APS-1 candidate gene was assigned to chromosome 21q22.3 by linkage analyses in patients with APS-1 from Finland. To examine the influence of ethnic and geographic differences on the location of the candidate gene locus, we studied 24 US patients with APS-1 by microsatellite marker typing, using five microsatellite markers, D21S49, PFKL, D21S171, D21S1903 and CD18, selected from chromosome 21q22.3. By allelic association analyses, the frequencies of allele number 5 for D21S171 and allele number 8 for D21S1903 were significantly higher in the 24 patients with APS-1 than in 33 controls (33/48 vs. 31/66, P = 0.0207, X2 = 5.35; 12/48 vs. 7/66, P = 0.0418, X2 = 4.15 respectively). The frequency of homozygosity for allele number 5 of D21S171 was also significantly higher in the patients than in controls, 15/24 vs. 9/33 (P = 0.0078, X2 = 7.07). Maximum lod scores detected for the five markers in nine families (containing 15 of the patients with APS-1) were: 2.384 for D21S49, 3.144 for PFKL, 3.506 for D21S171, 4.329 for D21S1903, and 1.130 for CD18. These results confirm the linkage of the candidate APS-1 gene to 21q22.3 in US APS-1 patients, and suggest that the candidate gene is located near the D21S1903 marker. The demonstration of the location of the APS-1 candidate gene to 21q22.3 in an out-bred heterogeneous patient population should promote the physical mapping of the responsible gene.

Alleles↗

Diabetic sera react with the glutamic acid decarboxylase molecule in a dimeric-oligomeric form.

Glutamic acid decarboxylase (GAD) is a major autoantigen in insulin-dependent diabetes mellitus (IDDM). This was initially identified as a 64-65 kD molecule according to migration in gels after immunoprecipitation from pancreatic islets. We studied the antigenicity of two different radiolabelled preparations of GAD, derived either by affinity purification from porcine brain and known to contain GAD 65 and GAD 67, or by expression from a cDNA for human GAD 65 by rabbit reticulocyte lysate (RRL). Radiolabelled immunoprecipitated pellets from the reaction of potent antisera to GAD from patients with IDDM were examined by autoradiography after SDS-PAGE under reducing or non-reducing conditions. Also, preparations of porcine brain GAD were "depleted' of GAD by exposure to antisera, and then similarly re-examined. Autoradiography of radiolabelled GAD either affinity purified from porcine brain, or expressed by RRL, showed that the immunoprecipitated protein migrated under non-reducing conditions according to a M(r) of approximately 110-130 kD, corresponding to dimeric forms of monomeric GAD of approximately 55-65 kD. Depletion by immunoprecipitation of this minor higher M(r) component from preparations of GAD left, in the supernatant, an abundance of GAD of M(r) 64-65 kD corresponding to monomer that was completely non-reactive with potent IDDM sera. We conclude that IDDM sera react with the GAD molecule in a dimeric (or oligomeric) form. Our findings have general connotations for self-tolerance and autoimmunity.

Animals↗

The distribution of HgCl2 in rat body and its effects on fetus.

Both female and male Wistar rats were dividend into 3 groups according to their body weights. The two test groups were given 75 micrograms/ml and 7.5 micrograms/ml HgCl, respectively, via drinking water, while the control group was given distilled water. Animals were mated after the male rats were treated for 12 weeks and female rats for 2 weeks. Blood samples from rats were taken on the 18th day of the pregnancy and the results showed that the HgCl2 was absorbed and stored in the blood in the form of inorganic mercury. The greatest amount of HgCl2 was found in the kidney. The placenta and the tests also had high mercury deposits. Fetuses showed low levels of HgCl2, thus, the placenta appears to serve as a barrier against HgCl2. No pathological changes were observed in the placenta, ovary, liver, and kidney in the female rats, but the epithelial cells of the rental proximal convoluted tubules were cloudy and swelling. The renal corpuscle showed no changes in the male rats. Pathological examination showed that a portion of the seminiferous epithelia were falling off, and the number of sperm released by the male was reduced. Examination by electron microscopy revealed that particulate fat was increased in sperm cells, which suggests that spermatozoa had been degenerated and were possibly one of the causes for the decreased rates of pregnancy in the exposed groups. No malformation of organs and bones in the rat fetuses were observed.

Animals↗

[Effect of ribavirin on NK activities of murine spleen cells in vitro].

The effects of ribavirin (1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide, RTC) on natural killer (NK) activities of murine spleen cells in vitro were studied by 125I-UdR isotope release assay. The results showed that RTC at the concentration of 5 mumol.L-1 significantly suppressed NK activities (P < 0.01), and a negative correlation between NK activities and RTC dosage was obtained. It also indicated that there was additive effect between the actions of RTC and dexamethasone (Dex, 1 mumol.L-1), and antagonism between the actions of RTC and interferon-gamma (100 U.ml-1). In addition, RTC at the concentration of 0.5 mumol.L-1 was found to be enough to reduce the product of interferon (P < 0.01) and interleukin-2 (P < 0.05) induced by Con A in murine spleen cells, and a negative dose-effect relationship was observed. Thus, one of the mechanisms of RTC in inhibiting NK activities seems to suppress the production of these lymphokines.

Animals↗