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Biomedical subjects

G Dai

Publications and source records attributed to G Dai.

At least 19 recordsLinked to original sources
PubMed2005

Stimulation of a rat uterine stromal cell line in culture reveals a molecular switch for endocrine-dependent differentiation.

Differentiation of uterine stromal cells is critical for the establishment of pregnancy. This study had two purposes: (i) to validate the use of the UIII rat uterine stromal cell model for investigating mechanisms underlying decidual cell differentiation, and (ii) to use this cell model to identify a molecular switch for cellular entry into the decidual cell differentiation pathway. Quiescent rat uterine stromal cells were transfected with a 500 bp segment of the decidual prolactin-related protein (dPRP) promoter ligated to a luciferase reporter gene. Cells were incubated in low-serum medium, or in low-serum medium containing progesterone (1 microM), estradiol 17-beta (10 nM), cholera toxin (10 ng/ml) and interleukin-11 (10 ng/ml). Protein extracts were collected 48 h later and luciferase was measured in the cellular lysates. Cholera toxin and interleukin-11 stimulated luciferase expression (P< 0.05) and addition of sex steroids further increased (P< 0.05) dPRP promoter activity. Stromal cells did not proliferate (P< 0.05) under differentiation conditions. Deletion analysis of the dPRP promoter revealed maximal luciferase expression between -250 and -500 bp relative to the transcription start site. Comparison of cyclin E/Cdk2 activity between proliferating and differentiating cells showed a 3-fold increase (P< 0.05) at 12 h in differentiating cells. The results suggest that cyclin E/Cdk2 serves as a molecular switch for uterine stromal cell entry into the decidual cell differentiation pathway.

Animals↗
PubMed2004

Modern diagnostics in chronic myeloproliferative diseases (CMPDs).

According to the new WHO classification a group of chronic myeloproliferative diseases (CMPDs) were defined: chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia and hypereosinophilic syndrome (CEL/HES), polycythemia vera (PV), chronic idiopathic myelofibrosis (with extramedullary hematopoiesis, CIMF), essential thrombocythemia (ET), and so called CMPD/unclassifiable. As clinical features and laboratory findings differ widely between these diseases several diagnostic approaches are mandatory at diagnosis for classification and are needed also for follow up studies, especially for the measurement of minimal residual disease (MRD). We here outline the laboratory set up at diagnosis and during follow up in CMPDs with specific focus on the respective therapeutical consequences. Only by using a comprehensive diagnostic panel including cytomorphology, cytogenetics, and molecular genetic methods establishing the correct diagnosis, optimizing treatment as well as evaluating treatment response is possible in CMPDs today.

Blood Cell Count↗
PubMed2004

Enhancement of murine IgE antibody detection by IgG removal.

RATIONALE: Although animal models for the study of allergic reactions are desirable, the use of mice has been hindered by the lack of sufficiently sensitive in vitro immunoglobulin epsilon (IgE) antibody assays. The aim of this study was to enhance IgE antibody measurements by immunoglobulin gamma (IgG) depletion. METHODS: Seven- to eight-week-old female mice of four strains (C3H/HeJ, CBA/J, C57Bl/6J, and Balb/c) were immunized (20 mice/group) with shrimp or peanut extracts using Al(OH)(3) as adjuvant. Following immunization, animals were sacrificed by exsanguination and the sera of each group pooled. Initial measurements of IgE antibody levels by enzyme-linked immunosorbent assay (ELISA) were relatively low; IgG and IgE reactivity patterns by immunoblot were similar. Thus, sera from shrimp or peanut immunized mice were depleted of IgG (absorbed 3-6 times with immobilized protein G) and then tested for IgE antibody to shrimp or peanut allergen. RESULTS: A 3- to 5-fold increase in IgE antibody reactivity as measured by ELISA was demonstrated when >80-90% of the IgG was removed. This increase in detection of allergen-specific IgE occurred in sera from all mouse strains and to all allergens tested. In addition, reactivity of IgE antibodies to peanut or shrimp allergens by immunoblot increased visually approximately 4- to 10-fold. CONCLUSIONS: These studies indicate that allergen-specific IgG antibodies, which may be in more than 100-fold excess to IgE antibodies, interferes with detection of allergen-specific IgE, probably by competitive binding to allergenic epitopes. Substantial depletion of IgG antibodies (>80%) result in a significant increase in the sensitivity of the antibody measurements.

Allergens↗
PubMed2003

The estimation of absorbed doses received by a victim of a Chinese radiation accident.

The aim of this work was to estimate absorbed doses received by a victim of the radiation accident with a 60Co source in Henan province, China. With a Monte Carlo stochastic simulation method, an estimation method for doses to the radiation accident victim was made. It utilised a mathematical model of adult man (MIRD) and a relative applied computer program was developed. By means of the simulated conditions of the accident, the absorbed doses to the victim's main organ and total body doses were estimated. The results estimated by our Monte Carlo method are close to those of experimental simulation measurement of the accident. With its convenience and rapidity, this method will be valuable for radiation dose reconstruction for victims in radiation accidents.

Adult↗
PubMed2002

Prolactin family miniarray: a tool for evaluating uteroplacental-trophoblast endocrine cell phenotypes.

The prolactin family represents a group of hormones and cytokines that participate in the control of maternal and fetal adaptations to pregnancy. The aim of this study was to develop a simple assay for monitoring patterns of prolactin family gene expression in rats and mice. Prolactin family cDNAs were spotted on to nylon membranes. Total RNA was extracted from tissues or cells. cDNAs were generated, radiolabelled using reverse transcriptase, and used as probes to hybridize with the prolactin family miniarrays. Pituitary, uterine decidual tissue and placenta each expressed a unique profile of prolactin family members. Placental tissues exhibited regional- and temporal-specific patterns of expression. Prolactin family gene expression differed markedly in mid-pregnant versus late gestation placental tissues and between the junctional and labyrinthine zones of the chorioallantoic placenta. Marked changes in prolactin family gene expression were also observed in cultured trophoblast cells undergoing differentiation. In conclusion, the prolactin family miniarray assay is an effective method for evaluating the endocrine phenotype of the uterus, placenta and trophoblast cells.

Animals↗
PubMed2001

[Influence of human cytomegalovirus infection on the expression of HOXB5, HOXB6, HOXB7, and HOXB8 genes in gliomaous cells].

OBJECTIVE: The expressions of HOXB5, HOXB6, HOXB7, and HOXB8 genes of U251 cell infected by human cytomegalovirus and/or treated with all trans-retinoic acid(ATRA) were detected by semi-quantitative RT-PCR. The results were that U251 cell did not express HOXB5, HOXB6, and HOXB8 but expressed HOXB7 without infecting HCMV and/or being treated with ATRA. After infected with HCMV and/or treated with ATRA, the expressions of HOXB7 and HOXB8 were up-regulated and the expression of HOXB7 lasted the fourth generation, while HOXB8 was up-regulated only in the second generation. These results indicate that the abnormal expression of HOXB gene induced by HCMV may play an important role in the maldevelopment.

Brain Neoplasms↗
PubMed2001

Allocation of helper T-cell epitope immunodominance according to three-dimensional structure in the human immunodeficiency virus type I envelope glycoprotein gp120.

The specificity and intensity of CD4(+) helper T-cell responses determine the effectiveness of immune effector functions. Promiscuously immunodominant helper T-cell epitopes in the human immunodeficiency virus (HIV) envelope glycoprotein gp120 could be important in the development of broadly protective immunity, but the underlying mechanisms of immunodominance and promiscuity remain poorly defined. In this study, gp120 helper T-cell epitopes were systematically mapped in CBA/J and BALB/c mice by restimulation assays using a set of overlapping peptides spanning the entire sequence of the gp120 encoded by HIV strain 89.6. The results were analyzed in the context of the HIV gp120 structure determined by x-ray crystallography. One major finding was that all of the promiscuously immunodominant gp120 sequences are located in the outer domain. Further analyses indicated that epitope immunogenicity in the outer domain correlates with structural disorder in adjacent N-terminal segments, as indicated by crystallographic B-factors or sequence divergence. In contrast, the correlation was poor when the analysis encompassed the entire gp120 sequence or was restricted to only the inner domain. These findings suggest that local disorder promotes the processing and presentation of adjacent epitopes in the outer domain of gp120 and therefore reveal how three-dimensional structure shapes the profile of helper T-cell epitope immunogenicity.

Amino Acid Sequence↗
PubMed2001

Delay of intracellular calcium transients using a calcium chelator: application to high-throughput screening of the capsaicin receptor ion channel and G-protein-coupled receptors.

Whole-cell functional assays are often used for high-throughput screening (HTS) of molecular targets such as ion channels and G-protein-coupled receptors. A common method for assaying the activity of these membrane proteins is to measure the change in intracellular calcium concentration upon receptor stimulation. These changes in calcium concentration are typically transient and therefore not readily adapted to high-density plate formats used in HTS instruments. We have demonstrated that an intracellular calcium chelator, BAPTA, was able to delay by 5- to 20-fold and extend for several minutes the observed calcium signals initiated by extracellular calcium influx or release of calcium from intracellular stores. As examples, we used cells expressing a calcium-permeable ion channel, vanilloid receptor type 1 (the capsaicin receptor), and two G-protein-coupled receptors. These receptor-mediated increases in intracellular calcium concentration were measured by both fluorescence-based and luminescence-based detection methods. The use of an intracellular calcium chelator to delay calcium signaling should have wide application since it allows the measurement of the functional activity of any cellular receptor that signals through calcium. With this procedure, calcium fluorescence and luminescence whole-cell functional assays may be performed with standard laboratory pipetting and detection systems.

Calcium↗
PubMed2001

[Influence of human cytomegalovirus infection on the expressions of HOXB1, HOXB5, HOXB6, and HOXB9 genes in human embryo lung cells].

OBJECTIVE: To study the expressions of HOXB1, HOXB5, HOXB6, and HOXB9 genes in human embryo lung (HEL) cells and influence of human cytomegalovirus (HCMV) infection on the expressions of these genes. METHOD: The expressions of HOXB1, HOXB5, HOXB6, and HOXB9 genes were detected with semi-quantitative RT-PCR method. RESULTS: 1. HEL cells expressed HOXB5 and HOXB6 genes, but they did not express HOXB1 and HOXB9 genes. 2. After HCMV infection, HEL cell was induced to express HOXB9 gene, as the expression of HOXB6 gene was elevated. The expression of HOXB5 gene had no significant change. HOXB1 was still not expressed. 3. Treated with all-trans retinoic acid (ATRA), the expression of HOXB9 gene in HEL cells infected by HCMV was significantly increased. But during the advanced infect period, the expression of HOXB6 was significantly decreased. CONCLUSION: HCMV can induce abnormal expressions of HOXB6 and HOXB9 genes, which may play an important role in HCMV-induced abnormal embryogenesis.

Cells, Cultured↗
PubMed2001

Simultaneous antegrade/retrograde cardioplegia protects myocardium distal to a coronary occlusion: a study in isolated pig hearts.

This study was designed to assess the effects of simultaneous antegrade/retrograde cardioplegia (SARC) on myocardial perfusion and energy metabolism in the region supported by the occluded left anterior descending artery (LAD) in isolated pig hearts. It was found that injection of Gd-DTPA into the aorta during antegrade cardioplegia (AC) did not result in signal increase in the LAD region on T(1)-weighted images. During SARC, however, Gd-DTPA was detected in the LAD region with the contrast agent injected into the aorta and the coronary sinus (CS), respectively. This suggests that SARC delivered blood cardioplegia to the jeopardized myocardium through both arterial and venous perfusion routes. Moreover, localized (31)P spectra showed that occlusion of the LAD during AC resulted in severe ischemic changes in the LAD myocardium and the abnormal metabolic changes were completely abolished by use of SARC. Finally, recovery of myocardial contractile function during reperfusion in the hearts subjected to SARC was significantly better compared to those arrested with AC alone. It was concluded that the myocardium distal to a coronary occlusion can be fully protected by use of SARC.

Animals↗
PubMed2001

Effects of flow and energy metabolism on injury and Rb(+) uptake in pig hearts: an (87)Rb and (31)P NMR study.

In this work the roles of coronary flow (CF) and metabolism in Rb(+) (K(+) congener) uptake were studied. In isolated pig hearts the left anterior descending artery (LAD) was cannulated to maintain adequate perfusion of the LAD bed. Rb(+) loading was initiated and the LAD flow was either completely stopped (no flow (NF)) or reduced to 12% (low flow (LF)), or buffer was deoxygenated without change in flow (hypoxic flow (HYP)) for 2 h. CF through the LAD was then restored to normal, or perfusion was switched to oxygenated buffer. Serial (87)Rb MR images or localized (31)P spectra were acquired to compare the Rb(+) uptake and energetics in the left ventricular (LV) anterior (ischemic/hypoxic) and posterior (normal) walls. End-ischemic/hypoxic (87)Rb signal intensities in the anterior wall were higher and the fluxes were greater in the HYP and LF groups than in the NF group. Phosphocreatine and ATP decreased less significantly and recovered better in the HYP and LF groups. Upon reperfusion/reoxygenation, the HYP and LF groups showed higher (87)Rb signal intensities and smaller or no infarctions in the anterior wall compared to those in the NF group. Ischemia reduces Rb(+) uptake due to both flow limitations and metabolic inhibition of cellular transport. (87)Rb MRI has a potential for distinguishing necrotic and reversibly damaged tissue.

Animals↗
PubMed2001

Is deep hypothermia necessary for unilateral antegrade cerebral perfusion during circulatory arrest? A magnetic resonance study in a pig model.

OBJECTIVE: Localized (31)P magnetic resonance spectroscopy (MRS) was used to investigate whether unilateral antegrade cerebral perfusion (U-ACP) could maintain normal energy metabolism and intracellular pH (pHi) in both hemispheres of the brain during deep (15 degrees C) and moderate (28 degrees C) hypothermic circulatory arrest (HCA). METHODS: Eleven pigs were exposed to 120 min of U-ACP during HCA at 15 degrees C (group I, n=6) or 28 degrees C (group II, n=5), followed by 60 min of cardiopulmonary bypass (CPB) at 37 degrees C. Localized (31)P MR spectra were acquired every 30 min. Histopathology was performed at the completion of each experiment. RESULTS: MR recorded no changes in energy metabolites (phosphocreatine and ATP), or pHi during U-ACP in either group, and no significant differences were found in any of the energy metabolites or pHi between the left and right hemispheres. Histopathology showed no significant morphological changes in the neurons. CONCLUSIONS: During either deep or moderate HCA, unilateral ACP through the right axillary artery prevents ischemic events in both hemispheres of normal pig brains. Deep hypothermia may not be necessary when using U-ACP.

Animals↗
PubMed2001

Effect on myocardial perfusion of simultaneous delivery of cardioplegic solution through a single coronary artery and the coronary sinus.

OBJECTIVE: This study was to determine whether simultaneous antegrade-retrograde cardioplegia through a single coronary artery and the coronary sinus provides sufficient and homogeneous perfusion to the heart. METHODS: Simultaneous antegrade-retrograde cardioplegia was conducted in 7 isolated pig hearts through the coronary sinus in conjunction with the left anterior descending artery, the left circumflex artery, and the right coronary artery, respectively. The efficacy of simultaneous antegrade-retrograde cardioplegia for myocardial perfusion was assessed by monitoring the distribution of magnetic resonance contrast agent and measuring the effluent from the venting coronary arteries. RESULTS: Injection of contrast agent into a perfusing artery during simultaneous antegrade-retrograde cardioplegia resulted in increased image signal intensity not only in the territory of the perfusing artery but also in the areas normally served by the other 2 venting arteries (including the right ventricular wall). The myocardium in the territories of the 2 venting arteries was lightened with contrast agent given into the coronary sinus during simultaneous antegrade-retrograde cardioplegia. Myocardium in the perfusing artery territory and right ventricular wall remained dark. Moreover, a significant amount of effluent was collected from the venting arteries during simultaneous antegrade-retrograde cardioplegia: 4.7 to 7.8 mL/min from the right coronary artery; 10.5 to 17.7 mL/min from the left anterior descending artery; and 9.7 to 15.2 mL/min from the left circumflex coronary artery. CONCLUSIONS: Simultaneous antegrade-retrograde cardioplegia through a single coronary artery and the coronary sinus provides homogeneous perfusion to the entire heart. During simultaneous antegrade-retrograde cardioplegia, arterial flow supports its own designated myocardium, as well as adjacent myocardium normally served by the venting arteries; the arterial route also supports the right ventricular free wall when the right coronary artery is vented. Venous perfusion of simultaneous antegrade-retrograde cardioplegia mainly supports myocardium in the territories of the venting arteries and does not perfuse the right ventricular free wall. Blood flow delivered to myocardium normally supported by the venting arteries is believed to be sufficient to prevent ischemic injury.

Animals↗
PubMed2001

The effect of combined arterial hemodynamics on saphenous venous endothelial nitric oxide production.

INTRODUCTION: Evidence exists that an ideal bypass conduit should have a functional endothelial cell surface combined with mechanical properties similar to those of native arteries. We hypothesized that the effect of combined arterial levels of pulsatile shear stress, flow, and cyclic strain would enhance saphenous venous endothelial cell nitric oxide (NO) production, and that variations in these "ideal" conditions could impair this function. We studied NO production as a measure of endothelial function in response to different hemodynamic conditions. METHODS: Human adult saphenous venous endothelial cells were cultured in 10-cm silicone tubes, similar in diameter (5 mm) and compliance (6%) to a medium-caliber peripheral artery (eg, popliteal). Tube cultures were exposed to arterial conditions: a combined pressure (120/80 mm/Hg; mean, 100 mm/Hg), flow (mean, 115 mL/min) and cyclic strain (2%), with a resultant pulsatile shear stress of 4.8 to 9.4 dyne/cm2 (mean, 7.1). Identical tube cultures were used to study variations in these conditions. Modifications of the system included a noncompliant system, a model with nonpulsatile flow, and a final group exposed to pulsatile pressure with no flow. NO levels were measured with a fluorometric nitrite assay of conditioned media collected at 0, 0.25, 0.5, 1, 2, and 4 hours. Experimental groups were compared with cells exposed to nonpulsatile, nonpressurized low flow (shear stress 0.1 dyne/cm2) and static cultures. RESULTS: All experimental groups had greater rates of NO production than cells under static conditions (P <.05). Cells exposed to ideal conditions produced the greatest levels of NO. Independent decreases in compliance, flow, and pulsatility resulted in significantly lower rates of NO production than those in the group with these conditions intact (vs noncompliant P <.05, vs nonflow P <.05, and vs nonpulsatile P <.05). CONCLUSIONS: Our results show that in the absence of physiologically normal pulsatility, cyclic strain, and volume flow, endothelial NO production does not reach the levels seen under ideal conditions. Pulsatile flow and compliance (producing flow with cyclic stretch) play a key role in NO production by vascular endothelium in a three-dimensional hemodynamically active model. This correlates biologically with clinical experience linking graft inflow and runoff and the mechanical properties of the conduit to long-term patency.

Adult↗
PubMed2001

Contrast agent distribution in microvascular damage of infarcted pig myocardium.

PURPOSE: We determined whether reperfusion damage was sufficient to allow extravasation of a large molecular weight contrast agent into infarcted pig myocardium. MATERIAL AND METHODS: Five pig hearts were subjected to in situ occlusion of the left anterior descending coronary artery (2 h) followed by reperfusion (1 h). The hearts were excised and perfused in the Langendorff mode for ex vivo MR imaging. Polylysine-Gd-DTPA (50,000 Da) and Gd-DTPA (500-700 Da) were injected into the aorta (alternately) and followed by measurements of T1 relaxation and mean transit time (MTT). RESULTS: In the normal myocardium, MTT of Gd-DTPA (56.8+/-23.2 s) was significantly (p=0.02) longer than that of polylysine-Gd-DTPA (29.0+/-7 s). However, both normal and infarcted myocardium showed similar MTT (29.0+/-7.0 vs. 28.0+/-5.0 s, p>0.05) when using polylysine-Gd-DTPA. CONCLUSION: The results indicate that the permeability of capillaries to polylysine-Gd-DTPA was not significantly higher in infarcted regions of the myocardium compared to normal tissue. However, infarcted myocardium displayed an increased permeability to the small molecular weight Gd-DTPA. We conclude that microvascular damage may not be sufficient to allow the extravasation of polylysine-Gd-DTPA in infarcted myocardium.

Animals↗
PubMed2001

CordyMax Cs-4 improves steady-state bioenergy status in mouse liver.

OBJECTIVE: To evaluate effects of CordyMax Cs-4, a mycelial fermentation product of Cordyceps sinensis, on energy metabolism. DESIGN: An in vivo pharmacology study using 31P nuclear magnetic resonance (NMR) spectroscopy. SUBJECTS AND STUDY INTERVENTIONS: Adult male C57-BL/6 mice were given an aqueous extract of CordyMax, 200 or 400 mg/kg per day or placebo for 7 days. OUTCOME MEASUREMENTS: Using 31P-NMR spectroscopy to measure cellular triphosphates and inorganic phosphate, expressed as a ratio to a reference peak, and calculate tissue pH. RESULTS: Steady-state beta adenosine triphosphate (ATP) increased in the liver of mice that received CordyMax (200 or 400 mg/kg per day) for 7 days, by 12.3% +/- 0.8% and 18.4% +/- 0.9%, respectively, compared to placebo controls (both p < 0.001), suggesting a higher hepatic bioenergy status in CordyMax-treated animals. Hepatic inorganic phosphate (Pi) decreased by 24.5% +/- 0.9% and 17.6% +/- 1.7% in the two treatment groups, respectively, compared to placebo controls (p < 0.001). The ratio of beta-ATP:Pi increased by 47.7% +/- 1.6% and 41.4% +/- 2.4%, respectively, in the treatment groups (both p < 0.001 compared to placebo). After discontinuation of CordyMax for 7 days, beta-ATP and Pi returned towards baseline. CONCLUSION: CordyMax is effective in improving bioenergy status in the murine liver, suggesting a mechanism underlying the known clinical effectiveness of CordyMax in alleviating fatigue and improving physical endurance, especially in elderly subjects.

Adenosine Triphosphate↗