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Biomedical subjects

J Cao

Publications and source records attributed to J Cao.

At least 19 recordsLinked to original sources

Selection of a phage-displayed peptide recognized by monoclonal antibody directed blocking the site of hepatitis C virus E2 for human CD81.

The human CD81 (hCD81) molecule has been identified as a putative receptor for hepatitis C virus (HCV). HCV envelope glycoprotein 2 (E2) most likely plays a pivotal role in binding to host cells by interacting with the hCD81 molecule. In this study, a phage-displayed peptide library was used to select small peptides with anti-hCD81 monoclonal antibody JS-81. The output/input ratio of phages increased about 91 fold after the third round of selection. Eight of the 30 phage clones selected from the phage library showed specific binding to the anti-hCD81 by enzyme linked immunosorbent assay (ELISA). Competitive inhibition test further demonstrated that HCV E2 could significantly inhibit the binding of a positive phage clone to anti-hCD81 JS-81. Exogenous small peptide ATWVCGPCT contained by the positive phage clones showed aligned with the hCD81 sequence from 153-161 by sequence analyses. These results suggest that the selected ATWVCGPCT is a novel hCD81-like small peptide, which can block the binding site of HCV E2 for hCD81. It may be of further application on development of antiviral agents targeting the stage of HCV entry.

Amino Acid Sequence↗

Opiate withdrawal modifies synaptic plasticity in subicular-nucleus accumbens pathway in vivo.

Subiculum receives output of hippocampal CA1 neurons and projects glutamatergic synapses onto nucleus accumbens (NAc), the subicular-NAc pathway linking memory and reward system. It is unknown whether morphine withdrawal influences synaptic plasticity in the subicular-NAc pathway. Here, we recorded the field excitatory postsynaptic potential (EPSP) within the shell of NAc by stimulating ventral subiculum in anesthetized adult rats. We found that high frequency stimulation (HFS, 200 Hz) induced long-term potentiation (LTP) but low frequency stimulation (LFS, 1 Hz) failed to induce long-term depression (LTD) in control animals. However, behavioral stress enabled LFS to induce a reliable LTD (sLTD) that was dependent on the glucocorticoid receptors. Both LTP and sLTD were prevented by the N-methyl-d-aspartate receptor antagonist AP-5. After repeated morphine treatment for 12 days, acute withdrawal (12 h) impaired LTP but had no effect on sLTD; prolonged withdrawal (4 days) restored the LTP but impaired the sLTD. Remarkably, basal synaptic efficacy reflected by baseline EPSP amplitude was potentiated in acute withdrawal but was depressed in prolonged withdrawal. Thus, acute and prolonged opiate withdrawal may cause endogenous LTP and LTD in the subicular-NAc pathway that occludes the subsequent induction of synaptic plasticity, demonstrating adaptive changes of the NAc functions during opiate withdrawal.

Animals↗

Short-term dynamic change of gill copper in common carp, Cyprinus carpio, evaluated by a sequential extraction.

Dynamic changes in Cu speciation and its binding to fish gills were investigated by exposing common carp (Cyprinus carpio) to a 1 mg/1 Cu solution for 7 hours. Cu speciation in the bulk solution and fish gill microenvironment was calculated using general chemical equilibrium modeling. A sequential extraction procedure using distilled water, magnesium dichloride (1.0 mol/1), and acetic acid (10%) was used to characterize the Cu associated with the fish gills. Cu residual in the gill tissue was measured after the sequential extraction. Changes in total Cu concentration, pH, and dissolved organic carbon (Doc) in the bulk solution were recorded during the experimental period and calculated for the fish gill microenvironment. Cu-hydroxide species and Cu2 were dominant Cu species in both bulk solution and the fish gill microenvironment, whereas increased Cu-mucus was found in the fish gill microenvironment. DOC in the exposure medium, assumed to arise from mucus release, also increased and complexed Cu in solution. Forty-three percent of the Cu associated with the gills was readily water extractable, with an additional 22% exchangeable with Mg2+ or protons. Only 35% of the Cu accumulated within the gill tissues. The binding of Cu to the fish gills reached apparent equilibrium after 3 hours of exposure. Furthermore, the amount of water-extractable Cu within the gills showed significant correlation to the concentration of Cu predicted to be complexed with calculated free mucus in the gill microenvironment.

Animals↗

N-methyl-D-aspartate receptor-dependent long-term potentiation in CA1 region affects synaptic expression of glutamate receptor subunits and associated proteins in the whole hippocampus.

Long term potentiation in hippocampus, evoked by high-frequency stimulation, is mediated by two major glutamate receptor subtypes, alpha-amino-3-hydroxyl-5-methyl-4-isoxazole propionate receptors and N-methyl-D-aspartate receptors. Receptor subunit composition and its interaction with cytoplasmic proteins constitute different pathways regulating synaptic plasticity. Here, we provide further evidence that N-methyl-D-aspartate receptor-mediated long term potentiation evoked at hippocampal CA1 region of rats induced by high-frequency stimulation of the Schaffer collateral-commissural pathway in vivo is not dependent on N-methyl-D-aspartate receptor subunit NR2B. Applying semi-quantitative immunoblotting, we found that in the whole tetanized hippocampus, synaptic expression of the N-methyl-D-aspartate and alpha-amino-3-hydroxyl-5-methyl-4-isoxazole propionate receptor subunits (NR1, NR2A, glutamate receptor 1) and their associated partners, e.g. synaptic associated protein 97, postsynaptic density protein 95, alpha subunit of Ca2+/calmodulin-dependent protein kinase II, neuronal nitricoxide synthase, increased 180 min post-high-frequency stimulation. Moreover, phosphorylation of Ca2+/calmodulin-dependent protein kinase II at thr286 and glutamate receptor 1 at ser831 was increased 30 min post-high-frequency stimulation and blocked by N-methyl-D-aspartate receptor antagonists (AP-5 and MK-801). In sham group and controls, these changes were not observed. The expression of several other synaptic proteins (NR2B, glutamate receptors 2/3, N-ethylmaleimide sensitive factor) was not affected by long term potentiation induction. In hippocampal homogenates, the level of these proteins remained unchanged. These data indicate that N-methyl-D-aspartate receptor-dependent long term potentiation in CA1 region in vivo mainly affects the synaptic expression of glutamate receptor subunits and associated proteins in the whole hippocampus. The alteration of molecular aspects can play a role in regulating the long-lasting synaptic modification in hippocampal long term potentiation in vivo.

2-Amino-5-phosphonovalerate↗

Alteration of cell-cycle regulatory proteins in human oral epithelial cells immortalized by HPV16 E6 and E7.

E6 and E7 oncoproteins from high-risk human papillomavirus (HPV) can transform cells in tissue culture and induce tumors in vivo by abrogating the cell-cycle checkpoint. To investigate the impact of HPV16 E6 and E7 on the cell-cycle regulatory machinery in oral epithelial cells, normal human oral epithelial cells were transfected with HPV16 E6 and E7 open reading frames, and alterations in cell-cycle regulatory proteins in cells expressing HPV16 E6 and E7 were analyzed. E6 and E7 expression results in immortalization of oral epithelial cells. E7 inactivates retinoblastoma protein (Rb) by forming complexes with hypophosphorylated Rb in immortalized oral epithelial cells. P53 and P21 protein levels were increased in immortalized cells compared to normal primary oral epithelial cells. Cyclin D1-cell-cycle-dependent kinase 4 binary association is disrupted in immortalized oral epithelial cells. These results indicate that E7 plays an important role in abrogation of cell-cycle regulation in oral epithelial cells, with E6 having a smaller impact. This suggests that the pathogenesis of HPV in oral epithelial cells differs from that in cervical epithelial cells.

Blotting, Western↗

Acute stress and intravesical corticotropin-releasing hormone induces mast cell dependent vascular endothelial growth factor release from mouse bladder explants.

PURPOSE: Corticotropin-releasing hormone is typically released from the hypothalamus but it has proinflammatory effects outside of the brain, possibly through the activation of mast cells. These cells express corticotropin-releasing hormone receptors with selective secretion of vascular endothelial growth factor, which may be involved in the pathogenesis of painful bladder syndrome/interstitial cystitis. This condition is characterized by bladder inflammation and worsened by stress. We investigated the effect of intravesical corticotropin-releasing hormone and acute restraint stress on vascular endothelial growth factor release from mouse bladder explants and the role of mast cells. MATERIALS AND METHODS: The bladder of C57BL/6 mast cell deficient (W/W(v)) and normal congenic (+/+) female mice (Jackson Laboratories, Bar Harbor, Maine) at ages 10 to 12 weeks was catheterized using anesthesia. After emptying urine 1) normal saline or corticotropin-releasing hormone was introduced for 45 minutes, urine was collected and the mice were allowed to recover for 4 hours before sacrifice or 2) the mice were stressed by placing them in a restrainer for 4 hours before sacrifice and urine was collected 2 hours after stress. The bladder was removed 4 hours after stress and processed for corticotropin-releasing hormone immunohistochemical staining. In other experiments the bladder was removed, minced into 1 mm(2) pieces and cultured with or without corticotropin-releasing hormone overnight. Urine and medium were frozen for histamine, interleukin-6, tumor necrosis factor-alpha and vascular endothelial growth factor assay. RESULTS: Corticotropin-releasing hormone (100 nM) or acute restraint stress (4 hours) increased histamine release in urine and vascular endothelial growth factor release in medium without increasing interleukin-6 or tumor necrosis factor-alpha in the bladder explants of C57BL/6 or +/+ but not W/W(v) mice. No vascular endothelial growth factor, interleukin-6 or tumor necrosis factor-alpha was detected in urine before or after stimulation. Corticotropin-releasing hormone immunoreactivity was present in control bladders but it increased dramatically in the bladder of stressed mice. CONCLUSIONS: Intravesical corticotropin-releasing hormone and acute restraint stress induced mast cell dependent vascular endothelial growth factor release from bladder explants. These findings suggest that stress, corticotropin-releasing hormone, mast cells and vascular endothelial growth factor might participate in the pathogenesis of painful bladder syndrome/interstitial cystitis, which is worsened by stress, and provide for new therapeutic targets.

Acute Disease↗

Concentrations and tracking of listeria monocytogenes strains in a seafood-processing environment using a most-probable-number enrichment procedure and randomly amplified polymorphic DNA analysis.

Concentrations of environmental microflora and Listeria monocytogenes were monitored at multiple environmental locations within a seafood-processing facility over the course of 6 months. Concentrations of L. monocytogenes were determined using a most-probable-number (MPN) enrichment procedure. Two floor drains had persistent low concentrations of L. monocytogenes (0.03 to >1,100 MPN/cm2). In comparison, concentrations of the other organisms in the drain were much higher (heterotrophic plate count range of 10(5) to 10(8) CFU/cm2). Concentrations of environmental organisms (heterotrophic aerobic plate counts and counts of pseudomonads, Shewanella spp., Aeromonas hydrophila, and coliforms) were not correlated with concentrations of L. monocytogenes. The 178 confirmed L. monocytogenes isolates from the MPN procedure were further characterized by randomly amplified polymorphic DNA analysis. Sixteen different banding patterns were identified, and nine of the patterns were identified from samples collected on two or more collection dates. From all locations, banding type A was observed in 98 confirmed isolates (55%). Although present, L. monocytogenes was a relatively minor component in the ecosystem of the floor drains in this seafood-processing facility.

Animals↗

Changes in serum cardiac troponins following a 21-km run in junior male runners.

AIM: This study was designed to examine the appearance and clearance characteristics of serum cardiac troponin T (cTnT) and I (cTnI) in junior runners after an exhaustive 21-km run. The dependence of the alternations of the cardiac troponins on the runners' training status reflected in training years, running ability and physiological profile at ventilatory threshold (Th(vent)) was also examined. METHODS: Ten trained male adolescents (16.2+/-0.6 years) performed 21-km run and graded treadmill exercise with maximum effort on two different days. cTnT and cTnI immediately before and 2, 4 and 24 hours after exercise were measured in the 21-km trial. The parameters at Thvent including the running speed (Th(vent)-Speed) and VO2 (Th(vent)-VO2) were identified in the graded exercise trial. RESULTS: Serum cTnT and cTnI in 6 of the 10 subjects measured 4 hours after the 21-km run were above the myocardial injury cutoffs of 0.03 and 0.09 ng x mL(-1), respectively, but returned towards pre-exercise levels within 24 hours that were not in the same manner observed in the clinical situation of myocardial infraction. The 6 subjects' training status was generally lower than that of the rest 4. Further, subjects' 4-h serum cTnT and cTnI were negatively related to their training years, Th(vent)-Speed and Th(vent)-VO2, and positively related to their personal best in half- and full-marathon races (r2 = or > 36%, n = 10). CONCLUSIONS: Such findings implied that clinicians based the diagnosis of long-distance run-induced myocardial infraction in novice junior runners upon the transient postexercise elevations of cardiac troponins alone should be with caution.

Adolescent↗

Geostatistical analysis and kriging of Hexachlorocyclohexane residues in topsoil from Tianjin, China.

A previously published data set of HCH isomer concentrations in topsoil samples from Tianjin, China, was subjected to geospatial analysis. Semivariograms were calculated and modeled using geostatistical techniques. Parameters of semivariogram models were analyzed and compared for four HCH isomers. Two-dimensional ordinary block kriging was applied to HCH isomers data set for mapping purposes. Dot maps and gray-scaled raster maps of HCH concentrations were presented based on kriging results. The appropriateness of the kriging procedure for mapping purposes was evaluated based on the kriging errors and kriging variances. It was found that ordinary block kriging can be applied to interpolate HCH concentrations in Tianjin topsoil with acceptable accuracy for mapping purposes.

China↗

Modeling the dynamic changes in concentrations of gamma-hexachlorocyclohexane (gamma-HCH) in Tianjin region from 1953 to 2020.

A level IV fugacity model was used to simulate the dynamic changes of gamma-hexachlorocyclohexane (gamma-HCH) concentrations in environmental media in Tianjin, China. A similar model (level III) was previously used and validated under steady state conditions; this paper explores its dynamic behavior. Application of the level IV fugacity model has been validated using independently observed gamma-HCH concentrations in various media during the early 1980s and during 2001. Sensitivity analysis was conducted using coefficient-of-variation normalized sensitivity coefficients. The model was also subject to uncertainty analysis using Monte Carlo simulation. It was found that concentrations of gamma-HCH reached within 95% of their steady-state levels in all media after less than 15 years. Around one order-of-magnitude decreases in gamma-HCH concentrations in various media occurred between 1993 and 2001. We project that gamma-HCH concentrations will decrease another 1.7-1.9 orders of magnitude to reach 1.1 (0.9-1.2) x 10(-14), 2.7 (2.5-3.0) x 10(-10), 1.2 (1.1-1.3) x 10(-7), and 6.1 (5.4-6.8) x 10(-8) mol/m(3), in air, water, soil, and sediment, respectively, by 2020. The sensitivities and true uncertainty of the model are discussed.

Air Pollutants↗

Optical control of coherent lattice motions probed by femtosecond electron diffraction.

We report the study of laser-induced coherent lattice motions using femtosecond electron diffraction. The oscillations of Bragg peak positions associated with a damped lattice vibration along the surface normal were directly observed in real time and with sub-milli-angstrom spatial resolution. In addition, by using a pair of optical excitation pulses and varying their time delay and relative pulse intensities, we demonstrated the successful control of coherent lattice motions.

Letter↗

The role of protein synthesis and degradation in the post-transcriptional regulation of rat multidrug resistance-associated protein 2 (Mrp2, Abcc2).

Multidrug resistance-associated protein 2 (Mrp2, Abcc2), an organic anion transporter present in the apical membrane of hepatocytes, renal epithelial cells, and enterocytes, is postulated to undergo post-transcriptional regulation. We hypothesized that Mrp2 protein undergoes altered rates of protein synthesis or degradation consistent with different Mrp2 protein expression. We analyzed Mrp2 synthesis, expression, and degradation in control female, 19- and 20-day pregnant, and pregnenolone-16alpha-carbonitrile (PCN)-treated rats using in vivo metabolic-labeling studies with [35S]cysteine/methionine or [14C]NaHCO3, polysomal distribution analyses and ribonuclease protection assays (RPA). Mrp2 protein was significantly increased in rats treated with PCN for 2 days but significantly decreased in 19-day pregnant rats relative to controls; no significant differences were observed in Mrp2 mRNA expression among these groups. The measured half-lives of 14C-labeled Mrp2 in control, pregnant, and PCN-treated rats were 27, 36, and 22 h, respectively, and were not significantly different. The rate of incorporation of 35S into Mrp2 was highest in PCN-treated rats. Polysomal distribution analysis of Mrp2 mRNA was consistent with increased Mrp2 protein synthesis after PCN treatment. The major transcription-initiation site for rat liver determined by RPA was -98 nucleotides (nt), with other start sites observed at -213, -163, -132, and -71 nt; use of transcription sites did not differ among the groups. Differences in the degradation of Mrp2 protein cannot explain the post-transcriptional regulation of Mrp2 in control, pregnant, and PCN-treated rats. Rather, the observed difference in protein synthesis suggests an intrinsic role for the translational regulation of rat Mrp2 protein.

ATP-Binding Cassette Transporters↗

Polycyclic aromatic hydrocarbons in dustfall in Tianjin, China.

Atmospheric dustfall samples from 23 locations in Tianjin, China, were collected and analyzed for 16 polycyclic aromatic hydrocarbons (PAHs) classified by the Environmental Protection Agency as priority pollutants from March 2002 to March 2003. SigmaPAH16 (sum of 16 PAH compounds) concentrations in the dustfall collected during heating season ranged from 2.5 to 85.5 mug/g, while that during the non-heating season varied from 1.0 to 48.2 microg/g dry weight. The dominant components in the heating season included naphthalene, phenanthrene, fluoranthene, and chrysene, while naphthalene, fluorene, phenanthrene, and fluoranthene were dominant during the non-heating season. Compared with the non-heating season, the heating season was characterized by a higher fraction of high-molecular-weight PAHs with four to six rings with exception of the samples from the east industrial area. The east industrial area had more significant correlations between individual PAH compounds, and more discrete triangular components of three-, four-, five- and six-ring PAHs. No significant correlations were observed between the PAHs concentrations and total organic carbon (TOC) in the dustfall samples. The deposition fluxes of sigmaPAH15 (sum of 15 PAHs except naphthalene), sigmaPAH6 (sum of 6 carcinogenic PAHs recommended by IARC) and benzo[a]pyrene (BaP) from atmospheric deposition to the whole area were estimated as 1911, 196, and 53 microg/m2/year, respectively. The deposition rates for PAH compounds in the east industrial area were higher than those in the urban and rural areas. Furthermore, the deposition contribution of PAHs during domestic heating season in winter was not significant relative to the annual inputs.

Air Pollution, Indoor↗

Human exposure and health risk of alpha-, beta-, gamma- and delta-hexachlorocyclohexane (HCHs) in Tianjin, China.

The dynamic exposures to HCHs of individuals born between the years 1913 and 1993 in Tianjin have been simulated by connecting a fugacity model (IV) with a multimedia exposure model. Ingestion is the most important pathway for human beings to take up HCHs, and concentrations in the human body correlate with body weight changes. Accumulations of HCHs were derived assuming that the degradation in human body behaved linearly. The health risk of exposure to HCHs was measured using cancer risk and loss of life expectancy (LLE), and LLE was modified from its original definition to incorporate a dynamic calculation that takes variances in exposure into account. Monte-Carlo simulations were run to analyze the uncertainties of the model.

Age Factors↗

Contamination of rivers in Tianjin, China by polycyclic aromatic hydrocarbons.

Tianjin urban/industrial complex is highly polluted by some persistent organic pollutants. In this study, the levels of 16 priority polycyclic aromatic hydrocarbons (PAHs) were tested in sediment, water, and suspended particulate matter (SPM) samples in 10 rivers in Tianjin. The total concentration of 16 PAHs varied from 0.787 to 1943 microg/g dry weight in sediment, from 45.81 to 1272 ng/L in water, and from 0.938 to 64.2 microg/g dry weight in SPM. The levels of PAHs in these media are high in comparison with values reported from other river and marine systems. Variability of total concentrations of PAHs in sediment, water, and SPM from nine different rivers is consistent with each other. No obvious trends of total PAHs concentration variations were found between upstream and downstream sediment, water, and SPM samples for most rivers, which indicate local inputs and disturbances along these rivers. The spatial distributions of three-phase PAHs are very similar to each other, and they are also similar to those found in topsoil. However, their chemical profiles are significantly different from that of topsoil. The change of profiles is consistent with the different aqueous transport capability of 16 PAHs. Low molecular weight PAHs predomination suggests a relatively recent local source and coal combustion source of PAHs in the study area.

China↗

Gene expression spectra in human leukemia HL-60 cells treated with EGCG.

To decipher the molecular mechanism of EGCG induced HL-60 cell apoptosis, alterations of gene expression spectra in HL-60 cell line cells after treatment with EGCG were screened by cDNA chip, and analyzed with the GenePix 3.0 twice; and the cDNA chip results further identified by RT-PCR. Ninety-seven genes among the total 8398 (1.15%) showed consistent significant differential expression in the duplicated cDNA chip assessments. Thirty-nine genes (40.2%) were up-regulated and 58 genes (59.8%) were down-regulated; and the randomly selected four performed RT-PCR results agreed with the cDNA chip data. The results suggest that the apoptosis of HL-60 cells induced by EGCG is a progressive transformation process regulated by a variety of genes.

Base Sequence↗

Application of the comet and micronucleus assays to the detection of B[a]P genotoxicity in haemocytes of the green-lipped mussel (Perna viridis).

Green-lipped mussels (Perna viridis) were exposed to water-borne benzo[a]pyrene (B[a]P) at nominal concentrations of 0, 0.3, 3 and 30 microg l(-1) for up to 12 days, and both the relative levels of DNA strand breaks (assessed using an alkaline comet assay) and the proportion of micronucleus (MN) formation were monitored in mussel haemocytes at days 0, 1, 3, 6 and 12. The results of the comet assay indicated that an increase in the proportion of strand breaks occurred generally with increasing B[a]P concentration, but a significant decrease in the levels of DNA damage was observed after exposure for 12 days at all concentrations tested, suggesting that the patterns of changes in the levels of DNA strand breakage can be explained by the threshold dependent DNA repair theory. Moreover, the relatively slow development and recovery of the DNA damage response in mussel haemocytes in comparison with previous findings utilizing P. viridis hepatopancreas suggests that the response of DNA alteration upon exposure to B[a]P may be tissue-specific in this species. Monitoring the frequency of micronucleus development in mussel haemocytes indicated both dose- and time-response relationships within the exposure period. Furthermore, the levels of DNA strand breakage correlated well with the levels of micronucleus induction, suggesting a possible cause and effect relationship between the two damage types. We suggest that DNA strand breakage and micronucleus formation in mussel haemocytes can potentially be used as convenient biomarkers of exposure to genotoxicants in the marine environment.

Analysis of Variance↗

Use of sequential ASE extraction to evaluate the bioavailability of DDT and its metabolites to wheat roots in soils with various organic carbon contents.

An accelerated solvent extraction (ASE) procedure using water, n-hexane and a mixture of n-hexane and acetone as solvents in sequence was developed and tested to evaluate the bioavailability of DDT and its metabolites including p,p'-DDT, o,p'-DDT, p,p'-DDE, and p,p'-DDD (SigmaDDTs) to wheat uptake from soils characterized by varied organic carbon contents. Results indicated that the extractability of SigmaDDTs with water was enhanced considerably in the presence of water soluble organic carbon (WSOC), while the amount of SigmaDDTs extracted with n-hexane was negatively correlated to the content of water insoluble organic carbon (WIOC). The interaction between SigmaDDTs and WIOC also reduced the bioavailability of the pesticides to wheat roots during uptake. There was a good positive correlation between the amount of SigmaDDTs extracted by n-hexane and the amount of SigmaDDTs accumulated in wheat roots, suggesting some potential for the use of the n-hexane ASE-extracted fraction as an indicator of SigmaDDTs' bioavailability to plant uptake. As such, the three sequentially extracted fractions may be viewed as representing the mobile, bioavailable, and fixed pools of SigmaDDTs in the soil.

Biological Availability↗