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Biomedical subjects

Hui Li

Publications and source records attributed to Hui Li.

At least 361 records · Page 20Linked to original sources

Facile synthesis of 1-thio-beta-lactoside clusters scaffolded onto p-methoxyphenyl, beta-D-galactopyranoside, beta-D-glucopyranoside, and lactoside.

The free-radical addition of 2,3,4,6-tetra-O-acetyl-beta-D-galactopyranosyl-(1-->4)-2,3,6-tri-O-acetyl-1-thio-beta-D-glucopyranose to the allyl ether functions of p-methoxyphenyl per-O-allyl-D-galactopyranoside, D-glucopyranoside, and lactoside provides a concise and effective route for synthesis of glycoside clusters, of use for exploring anti-metastatic activity.

Galactose↗

Tricarbocyanine cholesteryl laurates labeled LDL: new near infrared fluorescent probes (NIRFs) for monitoring tumors and gene therapy of familial hypercholesterolemia.

For monitoring low-density lipoprotein receptors (LDLr) in tumors and in livers of patients with familial hypercholesterolemia (FH) treated with gene therapy, a series of tricarbocyanine cholesteryl laurates were synthesized with the cholesteryl laurate moiety serving as the lipid-chelating anchor for low-density lipoprotein (LDL). One of these conjugates, TCL17, was successfully used to label LDL to give a new NIRF, TCL17-LDL. Ex vivo biological studies on an LDLr overexpressing tumor model, human hepatoblastoma G(2) (HepG(2)), confirmed that this NIRF were internalized selectively by the tumor and detected with high sensitivity by a low-temperature 3-D redox scanner.

Animals↗

Interactions that determine the assembly of a retinoid X receptor/corepressor complex.

The retinoid X receptor (RXR) is a key regulator in multiple signaling pathways because it can form either a homodimer with itself or a heterodimer with members of the class I nuclear receptors. The RXR-containing dimers regulate transcription by recruiting coactivators or corepressors to the target promoters. The binding of coactivators to RXR is mediated through a hydrophobic pocket formed in part by the C-terminal activation helix (AF-2). However, little is known about interactions of corepressors with RXR and its roles in transcriptional repression. Here we show that the repression activity of RXR correlates with its binding to the corepressor silencing mediator for retinoid and thyroid hormone receptors (SMRT). This intrinsic repression activity is masked by the AF-2 helix, which antagonizes SMRT binding. Inhibition of SMRT binding by the AF-2 helix requires specific amino acid sequences and the helical structure. Furthermore, the SMRT-binding site on RXR is independent of helix 11 but overlaps with the coactivator-binding pocket. On the basis of these results, we propose a structural model to help understand the molecular mechanism of corepressor recruitment by RXR.

Amino Acid Sequence↗

Experimental realization of quantum games on a quantum computer.

We generalize the quantum prisoner's dilemma to the case where the players share a nonmaximally entangled states. We show that the game exhibits an intriguing structure as a function of the amount of entanglement with two thresholds which separate a classical region, an intermediate region, and a fully quantum region. Furthermore this quantum game is experimentally realized on our nuclear magnetic resonance quantum computer.

Journal Article↗

Chromosomal autonomy of hMLH1 methylation in colon cancer.

Silencing of hMLH1 expression by aberrant hMLH1 promoter methylation accounts for the majority of sporadic colon cancers with microsatellite instability. We have previously shown hMLH1 silencing is biallelic and actively maintained. To study the mechanism of aberrant hMLH1 methylation, we assayed whether an hMLH1 methylated cell could transfer methylation and silencing to an exogenous hMLH1 promoter in somatic cell hybrids between hMLH1 methylated-silenced and hMLH1 unmethylated-expressing colon cancer cells. Conversely, we assayed whether these hybrids could reactivate expression of initially methylated and silenced hMLH1 alleles. Compellingly, within the hybrids each hMLH1 allele remained unchanged, retaining the expression status of its parental cell of origin. This chromosomal autonomy may not be simply determined by DNA methylation, as it is reasserted after experimentally forced demethylation of all hMLH1 alleles in the hybrids. Confirming findings included hMLH1 methylated cells being unable to methylate single transferred exogenous hMLH1 expressing chromosomes or transfected hMLH1 reporter constructs. hMLH1 silencing does not conform to either a dominant or recessive model, and is not determined by trans-acting factors differing between hMLH1 expressing or silenced genomes. We posit that hMLH1 methylation is dependent on and maintained by cis chromosomal marks, whose nature remains to be elucidated.

Adaptor Proteins, Signal Transducing↗

Comparative study of the active site caging of serine proteases: thrombin and factor Xa.

Bovine thrombin and human factor Xa were acylated at their active site selectively with inhibitors derived from the parent compound 4-guanidinophenyl (E)-4-diethylamino-2-hydroxy-alpha-methylcinnamate hydrochloride, 1b. Peptidyl side chains were attached to the phenol ring via amide connection, which served as a recognition motif in inhibiting different serine proteases. Upon irradiation with 366 nm light, the trans-cinnamate attached to the active-site serine isomerizes to the cis isomer which then rapidly lactonizes to release the free enzyme. The peptidyl side chain sequences specific for each serine protease were revealed via constructing and screening a library of homologous compounds. This methodology may be applied to other proteases. One application based on enzyme-specific, photoactivatable inhibitors is to isolate a designated active protease from a mixture of several proteases. Thus, a cinnamate inhibitor with a biotin moiety, 1d, was synthesized. A solution of enzyme-specific, biotinylated inhibitor was added into a mixture of proteases containing a target enzyme. The target enzyme was acylated at the active site and subsequently bore a biotin tail. An avidin column was used to separate the biotinylated enzyme from the unmodified ones, by a strong binding between biotin and avidin. After a brief irradiation on the avidin column, the retained enzymes were released from the biotin tag and eluted off the column. To demonstrate the idea, thrombin and factor Xa have been separated from each other by this strategy.

Acylation↗

Rapid method for the determination of total 5-methyltetrahydrofolate in blood by liquid chromatography with fluorescence detection.

A liquid chromatographic method is described for the determination of total 5-methyltetrahydrofolate (5-MTHF) in whole-blood samples. The method was applied to a survey of whole-blood total 5-MTHF levels of women at child-bearing age. To determine whole-blood total 5-MTHF content, a whole-blood sample was frozen and thawed to break red blood cells and the 5-MTHF polyglutamates were released and hydrolyzed into 5-MTHF monoglutamate by endogenous polyglutamates hydrolase in the plasma. In brief, an aliquot of 0.1 ml whole-blood sample was mixed with 0.3 ml 57 mmol/l ascorbic acid and incubated at 37 degrees C for 60 min, then diluted with 0.6 ml buffer solution (0.2 mol/l potassium phosphate dibasic and 30 mmol/l mercaptoethanol, pH 8.5). After the sample was heated at 100 degrees C for 10 min and centrifuged, the supernatant was analyzed by reversed-phase liquid chromatography with fluorescence detection. The recoveries from spiked samples were from 95 to 105% with within-day and day-to-day relative standard deviations less than 6.5%. The detection limit was estimated to be 30 nmol/l based on three times the noise level (peak to peak). Application of the method to a survey of whole-blood total 5-MTHF levels of women at child-bearing age showed that the method was reliable and suitable for the determination of blood total 5-MTHF.

Chromatography, Liquid↗

Autoreactive B cells in the marginal zone that express dual receptors.

Allotype and isotype exclusion is a property of most lymphocytes. The reason for this property is not known but it guarantees a high concentration of a single receptor, and threshold numbers of receptors may be required for efficient positive and negative selection. Receptor editing compromises exclusion by sustaining recombination even after a functional receptor is formed. Consequently, B cells expressing multiple receptors arise. We have studied such B cells in which one of the two receptors is anti-self, and find that these partially autoreactive B cells accumulate in the marginal zone. The restriction of these cells in this location may help to prevent them from undergoing diversification and developing into fully autoreactive B cells.

Animals↗

Advanced glycation endproduct-induced calcium handling impairment in mouse cardiac myocytes.

Long-standing diabetes causes cardiovascular complications including direct cardiac muscle weakening known as diabetic cardiomyopathy. This is characterized by disturbances in both cardiac contraction and relaxation, which are maintained by calcium homeostasis in cardiac cells. Our recent in vitro and in vivo studies have shown that advanced glycation endproducts (AGE) account for diabetic vasculopathy through their engagement of the receptor for AGE (RAGE). Here we show that AGE and RAGE may directly affect the myocardial Ca(2+) homeostasis. We created transgenic mice that overexpressed human RAGE in the heart and analyzed the Ca(2+) transients in cultivated cardiac myocytes (CM) from the RAGE-transgenic and non-transgenic control fetuses. RAGE overexpression was found to reduce the systolic and diastolic intracellular calcium concentration ([Ca(2+)](i)). Exposure to AGE caused a significant prolongation of the decay time of [Ca(2+)](i) in CM from control mice, and this response was augmented in CM from the RAGE transgenic mice. The results suggest that the AGE and RAGE could play an active role in the development of diabetes-induced cardiac dysfunction.

Animals↗

Site-specific integration of the double-mutation glucose isomerase (GIG138PG247D) gene in Streptomyces lividans and its stable expression.

A recombinant expression plasmid pYH12, containing the double-mutation glucose isomerase (GIG138PG247D, GI2) coding gene and its natural regulatory sequence, was constructed for site-specific integration in Streptomyces. The resulting plasmid was introduced into Streptomyces lividans TK54 by protoplast transformation and two apramycin-resistance (AmR) transformants, designated GY2 and BY7, respectively, were obtained further based on enzyme assays. These results for polymerase chain reaction (PCR), Dot blot, and recovery of cloned fragments from the transformant chromosome indicated that the GI2 gene was integrated into the S. lividans chromosome by site-specific recombination, and which was further verified by Southern blot. We found that the free form of plasmid pYH12 co-existing with the integrated form was present in S. lividans. SDS-PAGE analysis showed that the GI2 gene was expressed in S. lividans. The intracellular GI2 specific activity was 1.15 U/mg. The stability of integrants demonstrated that the cloned GI2 gene was stably integrated and expressed even in the absence of selective pressure.

Aldose-Ketose Isomerases↗

Uptake of trifluralin and lindane from water by ryegrass.

Understanding of the plant uptake of organic chemicals is essential to assessing contaminant mobility in the ecosystem, exposure to humans, and phytoremediation technologies. In this study, we measured the uptake of trifluralin and lindane from water by ryegrass as a function of uptake time for periods of 96 and 120 h, respectively. Trifluralin concentration in ryegrass increased sharply at the early stage of uptake and reached the maximum at 10 h, and then decreased with uptake time. 14C-labelled trifluralin uptake displayed a similar trend but a higher 14C-concentration than that of extracted parent compound, indicating metabolism and formation of bound residues following trifluralin uptake. Lindane concentration in ryegrass slowly increased with uptake time and approached a plateau, indicating minimal metabolism and formation of bound residues. The difference in the uptake characteristics of these two chemicals may be related to the differences in their lipophilicity, and chemical and biological reactivities. A two-compartment model accounting for the contributions of transpiration, metabolism and formation of bound residues to overall uptake was developed to assess the uptake kinetics. The model adequately described the uptake of trifluralin and lindane into ryegrass by providing the first-order rate constants of uptake, release, transpiration, and metabolism and formation of bound residues. These rate constants are used in calculating plant concentration factor (PCF). The ratios of trifluralin concentrations in ryegrass to its aqueous concentrations are between the PCF at thermodynamic equilibrium and the PCF at steady state, suggesting the utility of both PCF values.

Biological Availability↗

Postaxial polydactyly type A/B (PAP-A/B) is linked to chromosome 19p13.1-13.2 in a Chinese kindred.

Postaxial polydactyly is characterised by fifth digit duplications in hands and/or feet. Two phenotypic varieties have been described. In type A, the extra digit is well formed and articulates with the fifth or an extra metacarpal. A rudimentary extra fifth digit characterises type B. Mutations in the GLI3 gene are associated with postaxial polydactyly in some families and a second locus has been identified on chromosome 13 but the majority of cases remain unexplained. We report here a third locus for postaxial polydactyly on chromosome 19p13.1-13.2 in a large Chinese kindred with a combination of type A and B. Two-point linkage analysis showed the highest LOD score (Z(max) = 5.85 at theta; = 0 cM) at marker D19S221. Recombination events with markers D19S1165 and D19S929 define the critical region for this postaxial polydactyly locus to a region of 4.3 cM on the genetic map.

China↗

The effects of reverse atrial electrical remodeling on atrial defibrillation thresholds.

The implantable atrial defibrillator is a developing therapeutic option for paroxysmal atrial fibrillation, but shock related discomfort continues to be a limiting factor. To further characterize successful defibrillation, the relationship between reverse atrial electrical remodeling and internal atrial defibrillation thresholds in canines with chronic atrial fibrillation was examined. Testing was performed in 21 dogs. Chronic atrial fibrillation was induced in eight dogs by creating moderate mitral regurgitation and rapidly pacing the right atrium for > or = 6 weeks. The atrial fibrillation cycle length, atrial effective refractory period, refractory period dispersion, and internal atrial defibrillation thresholds were determined after establishment of chronic atrial fibrillation after 4 hours of sinus rhythm postcardioversion and 7 days of sinus rhythm postcardioversion. These measurements were then compared to a normal population of 13 dogs. The atrial defibrillation thresholds were 6.6 J (1.9-10.1 J) initially, 2.9 J (1.5-3.7 J) after 4 hours of sinus rhythm, and 0.9 J (0.4-1.3 J) after 7 days of sinus rhythm (P = 0.04). This decrease was associated inversely with the atrial effective refractory period (P < 0.03), and atrial fibrillation cycle length (P < 0.05), and with a decrease in atrial refractory period dispersion after 7 days of sinus rhythm (P = 0.04). These electrophysiological measurements reached normal population levels by 7 days. Atrial defibrillation thresholds decrease as atrial reverse electrical remodeling occurs and this reduction corresponds to increased atrial fibrillation cycle length, increased atrial refractoriness, and decreased refractory period dispersion.

Animals↗

Effects of aldosterone on biosynthesis, traffic, and functional expression of epithelial sodium channels in A6 cells.

The collecting duct regulates Na(+) transport by adjusting the abundance/activity of epithelial Na(+) channels (ENaC). In this study we have investigated the synthesis, degradation, endocytosis, and activity of ENaC and the effects of aldosterone on these processes using endogenous channels expressed in the A6 cell line. Biochemical studies were performed with a newly raised set of specific antibodies against each of the three subunits of the amphibian ENaC. Our results indicate simultaneous transcription and translation of alpha, beta, and gamma subunits and enhancement of both processes by aldosterone: two- and fourfold increase, respectively. The biosynthesis of new channels can be followed by acquisition of endoglycosidase H-resistant oligosacharides in alpha and beta subunits and, in the case of alpha, by the appearance of a form resistant to reducing agents. The half-life of the total pool of subunits (t(1/2) 40-70 min) is longer than the fraction of channels in the apical membrane (t(1/2) 12-17 min). Aldosterone induces a fourfold increase in the abundance of the three subunits in the apical membrane without significant changes in the open probability, kinetics of single channels, or in the rate of degradation of ENaC subunits. Accordingly, the aldosterone response could be accounted by an increase in the abundance of apical channels due, at least in part, to de novo synthesis of subunits.

Aldosterone↗

Characterization of the neurotrophic response to acute pancreatitis.

INTRODUCTION: Interesting preliminary data on changes in the neurotrophin system in various digestive diseases have recently begun to emerge. AIMS: To measure changes in messenger RNA (mRNA) levels of neurotrophins and to identify cell types expressing neurotrophins in the pancreas of rats with L-arginine-induced pancreatitis. METHODOLOGY: Rats were killed at time points from 2 hours to 4 weeks after the induction of pancreatitis, and responses were measured by assay. RESULTS: By RNase protection assay, ciliary neurotrophic factor (CNTF) mRNA expression showed a rapid response (sixfold increase over control) in the inflamed pancreas at 2 hours. The levels of mRNA expression of brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), neurotrophin-3 (NT-3), and neurotrophin-4 (NT-4) in the inflamed pancreas reached a peak at 1 week (2.5-fold, twofold, fourfold, and fivefold increase, respectively). By immunohistochemistry, immunoreactivity for all neurotrophins examined was observed in the islets of Langerhans in the control pancreas at all time points, but it was markedly reduced in the islets in the inflamed pancreas at 2 and 6 hours. Acinar and ductal cells, inflammatory cells, and neural elements were immunoreactive for those neurotrophins in the inflamed pancreas from 2 hours to 2 weeks. CONCLUSION: The temporal and spatial expression of neurotrophins in the course of experimental pancreatitis suggests that their upregulation is a critical component of the response of the pancreas to injury in this model.

Acute Disease↗

Activities of arginase I and II are limiting for endothelial cell proliferation.

Polyamines are essential for cell proliferation; therefore, we hypothesized that arginase I or arginase II activities, via production of ornithine for polyamine synthesis, may be limiting for proliferation of endothelial cells (EC). Bovine coronary venular EC stably transfected with a lacZ gene (lacZ-EC, control), rat arginase I cDNA (AI-EC), or mouse arginase II cDNA (AII-EC) were utilized to test this hypothesis. Cell-proliferation assays showed that EC proliferation was markedly increased in AI-EC and AII-EC compared with lacZ-EC. Expression of proliferating cell nuclear antigen was also enhanced in AI-EC and AII-EC. DL-alpha-difluoromethylornithine (DFMO), an irreversible inhibitor of ornithine decarboxylase, was used to establish that increased polyamine synthesis was involved in mediating the enhanced growth of AI-EC and AII-EC. Addition of 5 mM DFMO to the culture medium completely abolished the differences in cellular putrescine concentrations and reduced the differences in spermidine concentrations among AI-EC, AII-EC, and lacZ-EC. The DFMO treatment also prevented an increase in AI-EC and AII-EC proliferation compared with lacZ-EC. Addition of 10 and 50 microM putrescine dose-dependently increased AI-EC, AII-EC, and lacZ-EC growth to the same extent. These results demonstrate that either arginase isoform can potentially play a role in modulating EC proliferation by regulating polyamine synthesis.

Animals↗