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Biomedical subjects

W Dong

Publications and source records attributed to W Dong.

At least 55 records · Page 3Linked to original sources

Attenuated allergic responses to house dust mite antigen in feed-restricted rats.

Caloric restriction has been shown to alter a broad range of immunological end points in both experimental animals and humans. The objective of this study was to investigate the effect of short-term moderate feed restriction (25% reduction) on allergic immune responses in Brown Norway rats. After 3 weeks of acclimation to their feed regimens, rats were sensitized and 2 weeks later challenged with house dust mite (HDM) antigen via intratracheal instillation. Feed restriction resulted in lower levels of antigen-specific IgE in serum and reduced antigen specific lymphoproliferative activity in pulmonary lymph nodes. Feed restriction also attenuated pulmonary inflammation, as evidenced by lower levels of lactate dehydrogenase and total protein, decreased infiltration of neutrophils and eosinophils, and reduced secretion of pro-inflammatory cytokine tumor necrosis factor (TNF)-[alpha] in bronchoalveolar lavage fluid. In addition, feed restriction decreased TNF-[alpha] secretion in serum and decreased mRNA expression of TNF-[alpha] and interleukin-6 in pulmonary lymph nodes. We conclude that feed restriction strongly dampened the allergic immune responses to HDM in rats and that this attenuation was associated with decreased expression and secretion of pro-inflammatory cytokines.

Animals↗

Comparison of two kinds of methods to determine the titer of recombinant retrovirus containing beta-globin gene based on G418 selection.

Four recombinant retrovirus (RV) vectors containing human beta-globin gene and regulatory elements were constructed. To determine the titers of recombinant RV from corresponding producer cell lines, we compared two kinds of method (the simple and the conventional) based on G418 resistance, which is conferred by neo gene of RV vector. The results demonstrated that the simple method shortened the selection period to 3 d instead of 10-12 d with the conventional method and reduced the amount of work; importantly, the titers determined by the simple method were not different significantly from those measured by the conventional method. It can be concluded that the simple method can be used to determine the titers of recombinant RV containing not only cDNA but also genomic DNA with introns and complex regulatory elements instead of the conventional method.

3T3 Cells↗

[Study on relationship of activated protein C resistance and recurrent spontaneous abortion].

OBJECTIVE: To investigate whether activated protein C resistance (APC-R) is associated with recurrent spontaneous abortion. METHODS: APC-R was detected by activated partial thromboplastin time (APTT) in the presence and absence of APC. Subjects included 40 nonpregnant parous women (control group) and 55 patients with recurrent spontaneous abortion. RESULTS: There was significant difference for APC-R between controls and the patients (P < 0.01). Among the study group, the prevalence of APC-R positive rate had no relation to the numbers of abortion. The positive rate of APC-R in patients with late abortion was significantly higher than those with only early abortion (38% and 12% respectively). CONCLUSION: APC-R is partly associated with recurrent spontaneous abortion, especially late abortion. It has no relation to the numbers of abortion.

Abortion, Habitual↗

[The capability of IL-1 in the induction of chemokine production by murine thymic epithelial cell (MTEC1) clones].

To analyze the capability of IL-1 and IL-6 in the induction of chemokine (CF) production by mouse thymus epithelial cell (MTEC1) clones, MTEC1 cells were cloned through one cell culture and individual cell clones were established in long term culture referred to as MTEC1-DW clones. The constitutive production of IL-1, IL-6 and CF by MTEC1-DW clones was evaluated and the patterns of the cytokine production determined. Addition of exogenous IL-1 or IL-6 or both to the cultures of those MTEC1-DW clones that are unable to produce CF, and incubated for 2 days, then, to assess the chemotactic activity in the cell culture supernatants (SNs). In the opposite, addition of anti-IL-1 mAb(s) to the cultures of those MTEC1-DW clones that can produce IL-1 and CF to neutralize secreted IL-1 then, to test chemotactic activity in the SNs after 2-day incubation. The results showed that in the MTEC1-DW clones which were unable to constitutively produce IL-1 or CF, addition of IL-1 could induce these cloned cells to produce CF with high chemotactic activity. By constrast, addition of anti-IL-1 mAb(s) to those MTEC1-DW clones that constitutively produce IL-1 and CF could significantly inhibit them to produce CF. IL-6 only exhibited weak activity in the induction of CF production by those cloned cells. Therefore, in the cytokine network regulation, CF production is mainly induced by endogenously produced IL-1 in MTEC1 cells.

Animals↗

[Image and quantity analysis of blood-gas in rabbit's artery and Na(+)-K(+)-ATPase in their lungs during PE-SWD treated by HFJV].

This study was made to gain an insight into the mechanism of high-frequency jet ventilation(HFJV) treatment for the pulmonary edema after seawater drowning(PE-SWD). PaO2, PaCO2 and oxygen saturation (SaO2) of the rabbit and Na(+)-K(+)-ATPase in the rabbit's lungs in three groups--PE-SWD group(PE-SWD-G), HFJV group (HFJV-G) and control group(CG), were measured and analysed by the blood-gas analyser and computer image system. The results showed that, after 100 minutes' HFJV, the PaO2, SaO2 and the activity of Na(+)-K(+)-ATPase in the lung capillary endothelial cells in HFJV-G were significantly higher than those in PE-SWD-G(P < 0.01 or P < 0.05). Three parameters of Na(+)-K(+)-TAPase(G1, D1 and D2) in HFJV-G almost returned to their values in CG. The authors suggest that the increase of PaO2 and SaO2 in the rabbit's artery blood in HFJV-G is closely related to the rehabilitation of Na(+)-K(+)-ATPase activity in the lungs after HFJV. The mechanism of successful HFJV treatment for PE-SWD is that HFJV can better correct hypoxemia and improve the rehabilitation of Na(+)-K(+)-ATPase activity in the rabbit's lungs.

Animals↗

[Study on the mechanism of pulmonary edema after seawater drowning in rabbit].

To study the mechanism of pulmonary edema after seawater drowning (PE-SWD), the indexes of blood-gas and acid-base in rabbits artery blood were measured by the blood-gas analyser. The activity of Na(+)-K(+)-ATPase, cytochrome oxidase(CYTO) and alkaline phospharase(ALP) in the lungs were measured and analysed by computer image system. C-fos mRNA and Fos protein in the lungs were respectively determined by situ hybrioization and immunohisto chemical techniques. The distribution of phospholipid and Ca2+ of rabbits lungs was quantitatively analysed by ultrastructural location method. The results showed that, five parameters of PaO2, oxygen saturation(SaO2), pH, actual bicarbonite(AB) and base excess(BE) and the activity of Na(+)-K(+)-ATPase and CYTO decreased remarkably in PE-SWD. Both c-fos mRNA and Fos protein expression in pulmonary epithelial cells in PE-SWD were significantly elevated compared with the normal controls(P < 0.01). The phospholipids products in the pulmonary alveolar type II epithelial cells were decreased, however, the Ca2+ precipitate pellets inside the lung capillary endothelial cells and the pulmonary alveolar type I and II epithelial cells increased obviously. The arthors suggest that the injuny action of the seawater, hypoxia and metabolic acidosis may be the mian three mechanisms of the pulmonary edema induced seawater drowning. The lowering activity of Na(+)-K(+)-ATPase and CYTO in the lungs and calcium overload in the cells are not only evil consequence resulted from above three factors, but also the importent causes leading to the worse of PE-SWD. The transmiting line of Ca(2+)-fos may be also a key link to bring about the worse of PE-SWD.

Acidosis↗

Residual oil fly ash exposure enhances allergic sensitization to house dust mite.

Epidemiological studies have shown an association between elevated levels of particulate matter air pollution and increased morbidity and hospital visits in asthmatics. Residual oil fly ash (ROFA) is a primary combustion particle containing sulfate and metals such as vanadium, nickel, and iron. In this study the effect of ROFA on sensitization to house dust mite (HDM) was examined in a Brown Norway rat model of pulmonary allergy. Rats were instilled via the trachea with 200 or 1000 micrograms ROFA 3 days prior to local sensitization with 10 micrograms HDM and were challenged with 10 micrograms HDM 14 days later. Immunological endpoints were examined at 2, 7, and 14 days after sensitization and at 2 and 7 days after challenge (16 and 21 days post-sensitization, respectively). Antigen-specific immunoglobulin E and associated immediate bronchoconstriction responses to antigen challenge were increased in the ROFA-treated groups compared with the HDM control group. Lymphocyte proliferation to antigen was enhanced at Days 7 and 21 in the bronchial lymphocytes of ROFA-treated groups. Bronchoalveolar lavage fluid (BALF) eosinophil numbers and lactate dehydrogenase were significantly increased in the 1000 micrograms ROFA group at Days 2 and 16, BALF total proteins were elevated at Days 2 and 7 in both ROFA-treated groups, and BALF interleukin (IL)-10 was elevated in the 1000 micrograms ROFA group at Day 2. These results suggest that ROFA has an adjuvant effect on sensitization to HDM.

Air Pollutants↗

NMR analysis of cardiac troponin C-troponin I complexes: effects of phosphorylation.

Phosphorylation of the cardiac specific amino-terminus of troponin I has been demonstrated to reduce the Ca2+ affinity of the cardiac troponin C regulatory site. Recombinant N-terminal cardiac troponin I proteins, cardiac troponin I(33-80), cardiac troponin I(1-80), cardiac troponin I(1-80)DD and cardiac troponin I(1-80)pp, phosphorylated by protein kinase A, were used to form stable binary complexes with recombinant cardiac troponin C. Cardiac troponin I(1-80)DD, having phosphorylated Ser residues mutated to Asp, provided a stable mimetic of the phosphorylated state. In all complexes, the N-terminal domain of cardiac troponin I primarily makes contact with the C-terminal domain of cardiac troponin C. The nonphosphorylated cardiac specific amino-terminus, cardiac troponin I(1-80), was found to make additional interactions with the N-terminal domain of cardiac troponin C.

Amino Acid Sequence↗

Prevalence of primary HIV drug resistance among seroconverters during an explosive outbreak of HIV infection among injecting drug users.

OBJECTIVES: This study examined the frequency of transmission of drug resistant HIV in the population of injecting drug users (IDU) in Vancouver, Canada during a period of particularly high virus transmission. DESIGN: All subjects enrolled in the Vancouver Injection Drug Users Study who seroconverted from HIV negative to positive status (n = 61) between December 1996 and February 1998 were eligible for analysis. The first seropositive sample from 57 individuals with plasma samples available was analyzed for resistance to antiretroviral agents by population based sequencing of the HIV protease and reverse transcriptase genes. METHODS: Plasma viral RNA was extracted and the viral reverse transcriptase and protease regions were amplified by nested reverse transcription-PCR. The presence of mutations associated with antiretroviral drug resistance was assessed by automated sequence analysis. RESULTS: Protease and reverse transcriptase sequences were successfully obtained from the 57 recent seroconverters. No cases of transmission of variants associated with significant resistance to protease inhibitors or nucleoside and non-nucleosides reverse transcriptase inhibitors were detected. CONCLUSION: The frequency of transmission of drug resistant HIV amongst these recently infected IDU is extremely low, with no protease or reverse transcriptase inhibitor resistant strains detected soon after seroconversion. The data provide no rationale for withholding treatment from this already marginalized population.

Anti-HIV Agents↗

A multisolution method of phase determination by combined maximization of entropy and likelihood. VI. The use of error-correcting codes as a source of phase permutation and their application to the phase problem in powder, electron and macromolecular crystallography.

The use of error-correcting codes as a source of efficient designs of phase permutation schemes is described. Three codes are used, all taken from the Bricogne BUSTER program [Bricogne (1993). Acta Cryst. D49, 37-60]: the Hamming [7, 4, 3], the Nordström-Robinson (16, 256, 6) and the Golay [24, 12, 8] or its punctured [23, 12, 7] form. These are used in a maximum-entropy-likelihood phasing environment to carry out phase permutation of basis-set reflections instead of the usual quadrant permutation or magic integer approaches. The use of codes in this way inevitably introduces some errors in the phase choices, but for most structures this is not significant especially when the gain in sampling efficiency is considered. For example, the Golay [24, 14, 8] allows the permutation of 24 centric phases in such a way that only 4096 phase sets are produced instead of 2(24) = 16777216, and one of these sets has, at most, only four wrong phases. The method is successfully applied to three powder diffraction data sets of increasing complexity, and with increasing degrees of overlap {Mg(3)BN(3), Sigma-2 ([Si(64)O(128)].4C(10)H(17)N) and the NU-3 zeolite}, a sparse electron diffraction data set for buckminsterfullerene, C(60), and the small protein molecule crambin at 3 Å resolution where 42 reflections are phased with a Uweighted mean phase error of 58.5 degrees.

Journal Article↗

Carboxypeptidase D is a potential candidate to carry out redundant processing functions of carboxypeptidase E based on comparative distribution studies in the rat central nervous system.

Post-translational processing is essential for the biological activation of many proteins and peptides. After precursor cleavage at specific single residues or pairs of basic residues by the proprotein convertases, the C-terminal basic residues are removed. Carboxypeptidase E was thought to be the only enzyme responsible. Recent studies with carboxypeptidase E-deficient mice, Cpe(fat)/Cpe(fat), indicated the existence of carboxypeptidase E-like carboxypeptidases, such as carboxypeptidase D. In order to define potential redundant functions in vivo, we compared the distributions of both carboxypeptidases in the rat central nervous system and selected endocrine tissues. Carboxypeptidase D messenger RNA was abundantly expressed in glial cells in the gray and white matter, while neurons in several brain regions, such as the piriform cortex, basolateral amygdala and hippocampus, also expressed high levels of carboxypeptidase D messenger RNA. Co-localization of carboxypeptidases E and D messenger RNAs was observed in many brain regions, the spinal cord and endocrine tissues. Immunohistochemistry showed the intracellular distribution of carboxypeptidase D with a perinuclear pattern. The extensive distribution of carboxypeptidase D in both glial and neuronal cells indicates the important role of carboxypeptidase D in peptide processing, possibly working together with furin, a ubiquitously expressed proprotein convertase. The co-localization of carboxypeptidases D and E suggests that carboxypeptidase D may, at least partially, compensate for carboxypeptidase E processing functions in Cpe(fat)/Cpe(fat) mice.

Adrenal Glands↗

Blood lead and blood pressure: evidence from the Health Survey for England 1995.

OBJECTIVES: To examine the relationship between blood lead and blood pressure (BP) and to estimate the possible effects of a decrease in blood lead on BP. METHODS: A 2-ml blood sample was collected from a sub-sample of those included in the Health Survey for England 1995, a cross-sectional survey of a nationally representative sample of the adult English population. Blood lead concentration was measured by atomic absorption spectrometry and three BP readings were taken under standardised conditions using the Dinamap 8100 monitor. Analyses were carried out using data on 2563 men and 2763 women aged 16 and over. RESULTS: In stepwise multiple regression analyses adjusting for various confounders--age, body mass index, smoking status, social class, region of residence and alcohol intake--blood lead was found to be significantly and positively associated with diastolic BP, and not systolic BP in men, but not in women. These findings were unaffected by the inclusion or exclusion of those on antihypertensive medication, by whether mean or median BP was used in the regression, or by the adjustment for alcohol consumption. A halving of currently prevalent blood lead levels is estimated to be associated with a decrease of between 0.8 to 1.1 mm Hg diastolic BP in men. CONCLUSION: These findings in the context of other published data are consistent with a small pressor effect of environmental lead levels on BP. They support recommendations for further efforts to reduce lead in the environment.

Adolescent↗

The scope for cardiovascular disease risk factor intervention among people with diabetes mellitus in England: a population-based analysis from the Health Surveys for England 1991-94.

AIMS: To examine the scope for cardiovascular disease risk factor intervention among diabetic patients in England was examined using data from the Health Surveys for England 1991-94. This evaluation included calculating the proportion who require lipid lowering therapy according to the Standing Medical Advisory Committee (SMAC) guidelines. METHODS: The Health Survey for England is an annual, nationwide household-based in which anthropomorphic data, blood pressure, lipids and details of cardiovascular risk factors are collected from households after random stratification for geographical and socio-economic factors within a population sample of 39639 adults, 970 (2.3%) diabetic subjects were identified. RESULTS: Overall, 51% of those with diabetes had hypertension (systolic blood pressure > or = 160 mmHg or a diastolic BP > or = 95 mmHg or being on antihypertensive therapy), 27% were obese (body mass index > or = 30 kg/m2) and 19% were current smokers. One-third of those with hypertension were untreated and less than one-half of those on treatment had their hypertension controlled to below 160/ 95 mmHg. More than one-quarter had poor glycaemic control (glycated Hb>11% or an HbA1c>7.5%). Of those aged <70 years, 29% required lipid lowering therapy according to the SMAC guidelines and almost all (94%) of these were not on treatment. An analysis showed that, because the SMAC guidelines do not require high density lipoprotein (HDL) cholesterol to be measured, their use is likely to result in substantial underestimation of the need for lipid lowering, particularly in diabetic women. CONCLUSION: In England, among those with diabetes there is considerable unmet need for cardiovascular risk factor intervention, particularly for hypertension and raised cholesterol. With this baseline established, future improvements can be monitored.

Adult↗

Generation of stable recombinant retroviruses containing the beta-globin genes linked to complex regulatory elements by using transient transfection.

A transient packaging method for a retroviral vector containing beta-globin sequences, mediated by Ca3(PO4)2 precipitation, generated stable recombinant beta-globin retroviruses with intact viral genomes. We suggest the use of this method as an alternative means to overcome the production of beta-globin encoding retroviruses with aberrant viral genomes using the routine packaging procedure.

Genetic Vectors↗

A recombinant multi-epitope, multi-stage malaria vaccine candidate expressed in Escherichia coli.

OBJECTIVE: To construct and evaluate a recombinant multi-epitope, multistage malaria vaccine candidate expressed in Escherichia coli (E. coli). METHODS: A hybrid gene (HGF) encoding several putative immunodominant T or T/B epitopes from MSP-1, MSP-2, Pf155/RESA of Plasmodium falciparum (P. falciparum) and two immune-stimulating epitopes from interleukin-1 and tetanus toxin was synthesized. Two copies of HGF and a copy of gene encoding Pattaroyo's Spf66 were connected together to construct a sandwich hybrid gene HGFSP. The gene was cloned into an expression vector pWR450-I for production of a fusion protein with beta-galactosidase. Efficacy of this vaccine candidate in inducing specific immunity against malaria parasites was evaluated. RESULTS: Immunization of different species of animals with purified recombinant peptide showed that the peptide was able to induce remarkable antibody response to the immunized peptide as well as falciparum malaria parasites. The epitopes included in the construct could induce antibodies against the intact parasite proteins as demonstrated by western blotting, indicating the epitopes retained their antigenicity in the new peptide construct. Antibodies from animals immunized with recombinant HGFSP peptide exhibited good ability in inhibition of the in vitro growth of malaria parasites, augmentation of phagocytosis of the parasites or infected RBC by phagocytes, and facilitation of antibody dependent cell mediated cytotoxicity to the cultured malaria parasites. CONCLUSION: The recombinant peptide seems to be a potential candidate which is valuable for further investigation.

Animals↗

[Abnormal dynamic electrocardiogram in patients with acute cerebral infarction].

OBJECTIVE: To assess the effect of lesions of insular cortex on cardiac damages in patients with cerebral infarction. METHODS: 24-hour dynamic electrocardiogram recordings was measured were performed in 74 patients with hemispheric cerebral infarction in the acute phase and in 39 control subjects. RESULTS: Supraventricular arrhythmias were significantly increased in groups of right insular infarction as compared with groups of lesions in other regions and with the control. A significantly increased rate of ST segment abnormality was found in groups of left insular infarction as compared with groups of lesions in other regions and with the controls. CONCLUSION: The effect of hemispheric cerebral infarction on heart function are mainly correlated with destruction of insular or regions adjacent to the insular. Increased rate of supraventricular arrhythmias is associated with right insular infarction. While increased rate of ST segment abnormality is associated with left insular infarction.

Aged↗

[The expression of Ras protein in human middle ear cholesteatoma].

OBJECTIVE: To evaluate the possible role of Ras protein in acquired middle ear cholesteatoma. METHODS: The specimens from the acquired middle ear cholesteatoma tissue of 22 cases and the normal external ear skin of 10 cases were examined by immunohistochemical S-P method and computer image analysis. RESULTS: All epithelial layers of cholesteatoma revealed a relatively abundant expression of Ras protein, among which 15 cases showed an inner plasma membrane expression, 5 cases a cytoplasm and membrane expression, and 2 cases cytoplasm expression only. In contrast, the normal external ear skin revealed a sparse distribution of Ras protein in the inner plasma membrane of epithelial cells with a more intensely staining in the basal layer. The integral absorbency of Ras protein in the two types of tissues were 0.870 and 0.463 respectively with a significant difference between them. CONCLUSION: There is an aberrant regulation of Ras protein in cholesteatoma. Ras protein plays a pivotal role during the aberrant growth and the differentiation of cholesteatoma.

Adolescent↗

Prodynorphin processing by proprotein convertase 2. Cleavage at single basic residues and enhanced processing in the presence of carboxypeptidase activity.

Endoproteolytic processing of the 26-kDa protein precursor prodynorphin (proDyn) at paired and single basic residues is most likely carried out by the proprotein convertases (PCs); however, the role of PCs at single basic residues is unclear. In previous studies we showed that limited proDyn processing by PC1/PC3 at both paired and single basic residues resulted in the formation of 8- and 10-kDa intermediates. Because PC2 is colocalized with proDyn, we examined the potential role of this convertase in cleaving proDyn. PC2 cleaved proDyn to produce dynorphin (Dyn) A 1-17, Dyn B 1-13, and alpha-neo-endorphin, without a previous requirement for PC1/PC3. PC2 also cleaved at single basic residues, resulting in the formation of the C-peptide and Dyn A 1-8. Only PC2, but not furin or PC1/PC3, could cleave the Arg-Pro bond to yield Dyn 1-8. Structure-activity studies with Dyn A 1-17 showed that a P4 Arg residue is important for single basic cleavage by PC2 and that the P1' Pro residue impedes processing. Conversion of Dyn A 1-17 or Dyn B 1-13 into leucine-enkephalin (Leu-Enk) by PC2 was never observed; however, Dyn AB 1-32 cleavage yielded small amounts of Leu-Enk, suggesting that Leu-Enk can be generated from the proDyn precursor only through a specific pathway. Finally, PC2 cleavages at single and paired basic residues were enhanced when carried out in the presence of carboxypeptidase (CP) E. Enhancement was blocked by GEMSA, a specific inhibitor of CPE activity, and could be duplicated by other carboxypeptidases, including CPD, CPB, or CPM. Our data suggest that carboxypeptidase activity enhances PC2 processing by the elimination of product inhibition caused by basic residue-extended peptides.

Amino Acid Sequence↗