Biomedical subjects

Qian Zhao

Publications and source records attributed to Qian Zhao.

49 records · Page 3Linked to original sources

PubMed2004

N-substituted pyrrole derivatives as novel human immunodeficiency virus type 1 entry inhibitors that interfere with the gp41 six-helix bundle formation and block virus fusion.

A recently approved peptidic human immunodeficiency virus type 1 (HIV-1) fusion inhibitor, T-20 (Fuzeon; Trimeris Inc.), has shown significant promise in clinical application for treating HIV-1-infected individuals who have failed to respond to the currently available antiretroviral drugs. However, T-20 must be injected twice daily and is too expensive. Therefore, it is essential to develop orally available small molecule HIV-1 fusion inhibitors. By screening a chemical library consisting of "drug-like" compounds, we identified two N-substituted pyrroles, designated NB-2 and NB-64, that inhibited HIV-1 replication at a low micromolar range. The absence of the COOH group in NB-2 and NB-64 resulted in a loss of anti-HIV-1 activity, suggesting that this acid group plays an important role in mediating the antiviral activity. NB-2 and NB-64 inhibited HIV-1 fusion and entry by interfering with the gp41 six-helix bundle formation and disrupting the alpha-helical conformation. They blocked a d-peptide binding to the hydrophobic pocket on surface of the gp41 internal trimeric coiled-coil domain. Computer-aided molecular docking analysis has shown that they fit inside the hydrophobic pocket and that their COOH group interacts with a positively charged residue (K574) around the pocket to form a salt bridge. These results suggest that NB-2 and NB-64 may bind to the gp41 hydrophobic pocket through hydrophobic and ionic interactions and block the formation of the fusion-active gp41 core, thereby inhibiting HIV-1-mediated membrane fusion and virus entry. Therefore, NB-2 and NB-64 can be used as lead compounds toward designing and developing more potent small molecule HIV-1 fusion inhibitors targeting gp41.

CD4 Antigens↗

PubMed2004

[Activation of adenylate cyclase influences the sensitivity of acute promyelocytic leukemia cell lines to ATRA].

OBJECTIVE: To explore the molecular mechanism of APL cell resistance to ATRA. METHODS: The ATRA sensitive and resistant APL cell lines, NB4 and NB4-R1, were used as in vitro models. The effects of specific inhibitors and activators of adenylate cyclase (AC) and phosphodiesterase (PDE) on ATRA-induced differentiation was evaluated by cell morphology, cell surface antigen expression and nitroblue-tetrazolium (NBT) reduction assays. RESULTS: SQ22536, a specific antagonist of AC, could dramatically block ATRA-induced NB4 cell differentiation. When ATRA + SQ22536 group compared with ATRA group, the positivity of CD11b decreased from (95.9 +/- 2.5)% to (60.3 +/- 7.1)%, while the A(540) in NBT reduction assay decreased from 0.585 +/- 0.092 to 0.170 +/- 0.028 (P < 0.05). Forskolin, an agonist of AC, could overcome the resistance of NB4-R1 cells to ATRA. When ATRA + forskolin group compared with ATRA group, the positivity of CD11b increased from (34.3 +/- 5.3)% to (94.6 +/- 2.4)%, while the A(540) in NBT reduction assay increased from 0.110 +/- 0.028 to 0.395 +/- 0.049 (P < 0.05). In contrast, the specific antagonist and agonist of PDE, 3-isobutyl-1-methylxanthine (IBMX) and calmodulin, exerted little impact on ATRA treatment. CONCLUSIONS: The defaults in the initiation of AC activation may contribute to the resistance to ATRA in some APL cells.

Adenine↗

PubMed2003

Determination of the HIV-1 gp41 fusogenic core conformation modeled by synthetic peptides: applicable for identification of HIV-1 fusion inhibitors.

Triggered by receptor binding of gp120, the human immunodeficiency virus type 1 (HIV-1) gp41 changes its conformation to a fusogenic six-helix bundle structure. In the present study, this core conformation modeled by the peptides derived from the gp41 N- and C-terminal heptad repeat regions was determined by fluorescence native polyacrylamide gel electrophoresis and size exclusion high-performance liquid chromatography (HPLC). Two previously described small molecule HIV-1 fusion inhibitors significantly blocked the six-helix bundle formation. It suggests that these biophysical techniques can be used in a novel way to study the conformational change of gp41 during virus entry into cells and to identify HIV-1 fusion inhibitors.

Amino Acid Sequence↗

PubMed2003

The GABA(A) receptor mediates the hypnotic activity of melatonin in rats.

The present investigation assessed whether hypnotic activity of melatonin was mediated by the GABA(A) receptor in rats. Electroencephalography (EEG) was measured in this experiment. Melatonin, at a dose of 10 mg/kg ip, showed a significant sleep-promoting effect in rats. Flumazenil (3.5 and 7 mg/kg), a specific antagonist of the benzodiazepine (BZP) recognition site on the GABA(A) receptor, and picrotoxin (2 and 4 mg/kg), the ligand of the picrotoxin site on the GABA(A) receptor, seemed to be devoid of intrinsic influence on each sleep parameter when used alone, but they significantly antagonized the melatonin-induced increase in total sleep time (TS), slow-wave sleep time (SWS) and paradoxical sleep time (PS), and the decrease in time to sleep onset (TSO) and wakefulness time (W). A significant interaction was shown between melatonin and flumazenil or picrotoxin. When bicuculline methiodide (2 and 4 mg/kg), a specific antagonist of the GABA binding site on the GABA(A) receptor, was used together with melatonin, the melatonin-induced increase in TS, SWS and PS, and the decrease in W were abolished. However, there was no interaction between melatonin and bicuculline methiodide on sleep parameters except PS. These results indicate that the hypnotic activity of melatonin may be linked to the GABA(A) receptor and mediated through the BZP recognition site, the picrotoxin site on the GABA(A) receptor and partially through the GABA binding site on the GABA(A) receptor.

Animals↗

PubMed2003

Automatic quantitation of HIV-1 mediated cell-to-cell fusion with a digital image analysis system (DIAS): application for rapid screening of HIV-1 fusion inhibitors.

Human immunodeficiency viruses type 1 (HIV-1) mediated cell-to-cell fusion plays an important role in HIV-1 spread from infected cells to uninfected cells and in HIV-1 cytopathogenesis. In the present study, we developed a convenient cell fusion assay using a computer-controlled digital image analysis system (DIAS) for automatic quantitation. Compared with a manual quantitative method, DIAS automatic method is less laborious, and more rapid. Furthermore, it is more objective and less dependent on the researchers' experience. This method has great potential to be developed further as a high-throughput screening assay for rapid screening of HIV-1 fusion inhibitors, for evaluating the activity of HIV-1 entry inhibitors and for studying the mechanism of action of anti-HIV-1 agents.

Cell Fusion↗

PubMed2003

Effect of the postictal state on visual-spatial memory in immature rats.

Postictal cognitive impairment following seizures can last from minutes to days and be disabling to the patient. The purpose of this study was to compare the behavioral features of seizures with postictal memory impairment in young seizure-naive rats and rats with a prior history of status epilepticus (SE) and examine the relationship between postictal EEG changes and cognitive recovery. Following training in the water maze to asymptote levels of learning, rats with a prior history of SE and seizure-naive rats had flurothyl-induced generalized seizures and time to recovery to baseline was then measured. Following generalized seizures rats had impaired performance in the water maze with the duration of the cognitive deficits exceeding the length of the seizure. There was not a close relationship between duration of cognitive impairment and either latency to onset of seizure or duration. The animal's neurological status was a factor in the duration of cognitive impairment following seizures; while there were no differences between SE and seizure-naive rats in latency to seizure onset or duration of the seizures, animals with a prior history of SE had a longer period of impairment following a seizure than animals without such a history. Postictal cognitive impairment was associated with changes in theta activity in animals with a prior history of SEs but not in seizure-naive animals. Caffeine, when administered following the seizure, reduced postictal cognitive impairment in a dose-dependent manner. This study demonstrates that duration of postictal cognitive impairment is not directly related to duration of the seizure. The neurological status of the animal is a determining factor in duration of postictal impairment.

Animals↗

PubMed2003

Development of a cell-based enzyme-linked immunosorbent assay for high-throughput screening of HIV type 1 entry inhibitors targeting the coreceptor CXCR4.

CXCR4, a coreceptor for human immunodeficiency virus type 1 (HIV-1) X4 virus, plays an important role in virus entry into the target cells. Antiviral agents that bind to CXCR4 are expected to inhibit HIV-1 entry. A cell-based enzyme-linked immunosorbent assay (ELISA) was developed utilizing an anti-CXCR4 monoclonal antibody, 12G5, and cells expressing CD4 and CXCR4, U373-MAGI-CXCR4(CEM) cells. Using this assay, we demonstrated that 12G5 specifically binds to the CXCR4-expressing cells, but not to CCR5-expressing cells and cells without CXCR4 and CCR5, consistent with the results obtained by using flow cytometry. The well-characterized CXCR4 antagonists, T22, T14012 (a downsized analog of T-22), and AMD3100, effectively inhibited 12G5 binding to CXCR4-expressing cells, while HIV-1 entry inhibitors targeting CD4 and gp41 as well as HIV-1 reverse transcriptase and protease inhibitors did not block the binding of 12G5 to U373-MAGI-CXCR4(CEM) cells. The prepared plates containing the fixed cells could be stored at -80 degrees C for at least 5 months without affecting the cell reactivity with 12G5, which enhances the convenience of this method. This suggests that the cell-based ELISA is specific, sensitive, convenient, rapid, and economic. With a robotic sample processing system, this assay can be used for high-throughput screening of HIV-1 entry inhibitors targeted to the HIV-1 coreceptor CXCR4.

Anti-HIV Agents↗

PubMed2003

[Purification of porcine pulmonary surfactant by CO2 supercritical fluid extraction].

OBJECTIVE: To develop a supercritical fluid extraction (SFE) method for purifying porcine pulmonary surfactant. METHOD: An orthogonal design with 4 factors and 4 levels was used to optimize the SFE conditions. RESULTS: The best extraction conditions were established as the following: pressure 30 MPa, temperature 50 degrees Celsius, dynamic extraction for 4 h with 0.2 ml ethanol as the modifier. CONCLUSION: The new method for purifying porcine pulmonary surfactant by SFE-CO2 can cause drastic reduction of organic solvents, being convenient to use and effective to purify.

Animals↗

PubMed2002

XTT formazan widely used to detect cell viability inhibits HIV type 1 infection in vitro by targeting gp41.

XTT can be metabolically reduced by mitochondrial dehydrogenase in viable cells to a water-soluble formazan product. Thus XTT has been widely used to evaluate cell viability and to screen anti-HIV agents and the cytotoxicity of these agents. The present studies demonstrated that XTT formazan derived from XTT in cell culture significantly inhibits the fusion of HIV-1-infected cells with uninfected cells. Synthetic XTT formazan effectively inhibited the replication of laboratory-adapted and primary HIV-1 isolates and cell-to-cell fusion with low cytotoxicity. It blocks the six-helix bundle formation between peptides derived from the N- and C-terminal heptad repeat regions of the gp41 ectodomain (designated N- and C-peptides, respectively). Analysis by a computer-aided docking program indicates that XTT formazan may bind to the highly conserved hydrophobic pocket on the surface of the central trimeric coiled coil of gp41. These results suggest that XTT formazan inhibits HIV-1 entry by targeting the alpha-helical coiled-coil domain of gp41. This small molecular nonpeptide antiviral compound can be used as a lead for designing more effective HIV-1 entry inhibitors targeting the fusion stage of HIV-1 infection. But because XTT formazan itself has anti-HIV-1 activity, caution should be exercised when XTT is used to evaluate HIV-1 infectivity.

Animals↗

PubMed2002

A stereoselective intramolecular 1,3-dipolar nitrone cycloaddition for the synthesis of substituted chromanes.

A stereoselective intramolecular 1,3-dipolar nitrone cycloaddition useful in the synthesis of chromanes is described. The reaction relies on the use of a chiral auxiliary on the nitrone partner. Key to the success of the reaction is the choice of auxiliary and the choice of Lewis acid catalyst. Utilizing an auxiliary with a pendant coordinating group, and Zn(OTf)(2) as the Lewis acid, diastereoselectivities up to 22:1 could be achieved.

Journal Article↗

PubMed2002

Risk assessment at hazardous waste-contaminated sites with variability of population characteristics.

Risk assessment is considered to be an effective scientific tool which enables decisionmakers to manage hazardous waste-contaminated sites in a cost-effective manner while preserving public health. However, the current risk assessment framework proposed by the US Environmental Protection Agency (US EPA) has limitations in addressing the true variability of population characteristics. This study proposed a methodology that is different from the existing framework by accounting for the true variability of population characteristics. The key differences of the proposed methodology and the existing framework are the (1) use of the transient exposure concentration; (2) use of the entire population rather than a representative ideal individual; (3) use of age- and gender-based population subgroups to represent population characteristics; (4) use of a population growth model to represent growth dynamics; and (5) presentation of risk through a risk profile with risk summarized through a single indicator, potential cancer incidences (PCI). The proposed methodology was applied in a ground water contamination scenario due to benzene to determine its applicability. The results of the study showed that age-based variability of population characteristics is important in predicting the population risk while gender played a small role. The existing US EPA methodology and its variation using age-independent variability of population characteristics overestimate the risk given by PCI substantially, and therefore, the decisions can lead to costly cleanup goals. Population risk is not a single value but a distribution due to the contribution from ditferent individuals of the exposed population. Hence, the decision criterion proposed in this study, PCI, is found to be a useful indicator to describe population carcinogenic risk to the society under a variety of conditions and scenarios.

Demography↗

PubMed2002

Peptide and non-peptide HIV fusion inhibitors.

Fusion of the HIV envelope with the target cell membrane is a critical step of HIV entry into the target cell. The HIV envelope glycoprotein gp41 plays an important role in the fusion of viral and target cell membranes and serves as an attractive target for development of HIV fusion inhibitors. The extracellular domain of gp41 contains three important functional regions, i.e. fusion peptide (FP), N- and C-terminal heptad repeats (NHR and CHR, respectively). The FP region is composed of hydrophobic, glycine-rich residues that are essential for the initial penetration of the target cell membrane. NHR and CHR regions consist of hydrophobic residues, which have the tendency to form alpha-helical coiled coils. During the process of fusion of HIV or HIV-infected cells with uninfected cells, FP inserts into the target cell membrane and subsequently the NHR and CHR regions change conformations and associate with each other to form a fusion-active gp41 core. Peptides derived from NHR and CHR regions, designated N- and C-peptides, respectively, have potent inhibitory activity against HIV fusion by binding to the CHR and NHR regions, respectively, to prevent the formation of the fusion-active gp41 core. C-peptide may also bind to FP, thereby blocking its insertion into the target cell membrane. One of the C-peptides, T-20, which is in the phase III clinical trials, has potent in vivo activity against HIV infection and is expected to become the first peptide HIV fusion inhibitory drug in the near future. However, this peptide HIV fusion inhibitor lacks oral availability and is sensitive to the proteolytic digestion. Therefore, it is essential to develop small molecular non-peptide HIV fusion inhibitors having a mechanism of action similar to the C-peptides. One of the approaches in identifying the inhibitors is to use an immunological assay to screen chemical libraries for the compounds that potentially block the interaction between the NHR and CHR regions to form a fusion-active gp41 core. In combination with computer-aided molecular docking techniques, the first active non-peptide HIV fusion inhibitor targeting gp41, ADS-J1, was identified. Other potential candidates of non-peptide HIV fusion inhibitors have also been identified using different approaches. It is expected that both peptide and non-peptide HIV fusion inhibitors will be developed as new classes of anti-HIV drugs, which will be used alone or in combination with HIV reverse transcriptase and protease inhibitors, for the treatment of HIV infection and AIDS.

Acquired Immunodeficiency Syndrome↗

PubMed2002

Comparative effectiveness of an essential oil mouthrinse and dental floss in controlling interproximal gingivitis and plaque.

PURPOSE: To compare the effectiveness of rinsing with an essential oil-containing antimicrobial mouthrinse with that of dental floss in reducing interproximal gingivitis and plaque in an unsupervised 6-month clinical trial designed in accordance with ADA Acceptance Program Guidelines. MATERIALS AND METHODS: 319 qualifying subjects, aged 18-63, were randomized into one of three groups: essential oil mouthrinse (Listerine Antiseptic); dental floss (Reach Dental Floss); or a negative control rinse. At baseline, subjects received a complete oral soft tissue examination and scoring of the Modified Gingival Index (MGI), modified Quigley-Hein Plaque Index (PI), and bleeding index (BI). Following a complete dental prophylaxis and receiving flossing or rinsing instructions, subjects started on their respective regimen. They continued on their assigned regimen unsupervised at home, in addition to toothbrushing, and were reexamined at 3 and 6 months. The treatment groups were compared with respect to baseline demographic and clinical variables. The primary efficacy variables were mean interproximal MGI and PI at 6 months. Intergroup differences at 3 and 6 months were tested using a one-way analysis of covariance model with treatment as a factor and the respective baseline value as the covariate. In addition, the essential oil mouthrinse was compared to floss for interproximal gingivitis reduction using "at least as good as" statistical criteria. RESULTS: 301 subjects were considered evaluable. There were no statistically significant differences among the 3 groups at baseline, with the exception of the essential oil mouthrinse group having significantly fewer AfroAmerican subjects than the other two groups. For the interproximal MGI, the essential oil mouthrinse and flossing were both significantly more effective than the negative control (P < 0.001) at 3 and 6 months. The essential oil mouthrinse was shown to be "at least as good as" dental floss for the control of interproximal gingivitis. For the interproximal PI, the essential oil mouthrinse was significantly more effective than the negative control at 3 and 6 months (P < 0.001) while flossing was significantly more effective than the negative control at 3 months (P < 0.05) but not at 6 months. The essential oil mouthrinse was significantly more effective than floss (P < 0.001) at both these time periods.

Adolescent↗