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Biomedical subjects

Jun Li

Publications and source records attributed to Jun Li.

At least 289 records · Page 16Linked to original sources

Detection of T cell memory to measles virus in experimentally infected rhesus macaques by cytokine flow cytometry.

A low, average frequency (0.61%) of measles virus (MV)-specific CD4 and CD8+ T cells was detected in rhesus monkeys experimentally infected with or vaccinated against MV. Both IFN-gamma and TNF-alpha positive T cells were visualized by flow cytometry. However, the conditions of short-term culture and stimulation to detect MV-specific T cells required significant modifications from a previously established method that reliably detects T cells in rhesus monkeys persistently infected with SIV. Both whole viral antigen and short synthetic peptide pools were an adequate antigenic stimulus. MV-specific T cells were detectable up to 11 years after exposure to the virus, although we cannot rule out possible subclinical re-exposure of the monkeys to vaccine virus during this time. Thus, flow cytometric methods can permit mechanistic studies of antigen-specific memory T cell dynamics following an acute viral infection in a primate model.

Animals↗

Reliability of diffraction enhanced imaging for assessment of cartilage lesions, ex vivo.

OBJECTIVE: The assessment of articular cartilage integrity is of value for the detection of early degenerative joint disease in both the clinical and the research settings. It was the purpose of this study to determine the accuracy and reliability of identifying articular cartilage defects through Diffraction Enhanced Imaging (DEI), a high contrast radiographic imaging technique. DEI provides two new sources of image contrast to radiography: refraction and scatter rejection, besides the absorption of conventional radiography. DESIGN: Cadaveric tali were DEI imaged in the anterior-posterior position at the National Synchrotron Light Source. Two independent observers provided gross score evaluations (on a five point scale) of the trochlear surfaces. The DEI image of each trochlear surface was then graded (on a five point scale) by two additional independent observers who were blinded with regard to the gross evaluation of the articular surfaces. Inter-observer agreement for DEI grades was assessed with the weighted kappa statistic. Correlation of diffraction enhanced image score to the gross score was assessed with Spearman correlation coefficient. RESULTS: The defects of articular cartilage of talar trochleae could be visualized through DEI. The Spearman correlation of gross grades with DEI grades on the 165 talar regions for observers 1 and 2 were 0.91 and 0.91, respectively. The overall weighted kappa value for inter-observer agreement was 0.93, thus considered high agreement. CONCLUSIONS: DEI is accurate and reliable for detection of articular cartilage defects ex vivo. Even early stages of degeneration of cartilage can be visualized with this high contrast technique. Future studies will focus on the application of DEI to the identification of such lesions in vivo.

Aged↗

Phragmalin-type limonoids from the mangrove plant Xylocarpus granatum.

Five new phragmalin-type limonoids, xyloccensins Q-U (1-5), along with xyloccensin P were isolated from the stem bark of the mangrove plant Xylocarpus granatum. Their structures were elucidated on the basis of extensive 1D and 2D NMR as well as IR and MS spectroscopic data analyses.

Limonins↗

Polyhydroxylated steroids from the soft coral Sinularia dissecta.

A repeated silica gel column chromatography followed by HPLC purification on the methanol extract of marine soft coral Sinularia dissecta, resulted in the isolation of fifteen polyhydroxylated steroids (1-15), involving six new C-18 functionalized steroids, 3beta-acetoxy-1alpha,11alpha-dihydroxygorgost-5-en-18-oic acid (1), gorgost-5-en-1alpha,3beta,11alpha,18-tetrol (2), 18-acetoxy-1alpha,3beta,11alpha-trihydroxygorgost-5-ene (3), 24(S)-3beta-acetoxy-1alpha, 11alpha-dihydroxyergost-5-en-18-oic acid (4), 24(S)-ergost-5-en-1alpha,3beta,11alpha,18-tetrol (5), and dissectolide (6). The structures of the new compounds were determined on the basis of extensive spectroscopic data (IR, MS, (1)H and (13)C NMR, HMQC, HMBC, and NOESY) analysis. Compound 6 was found as an unusual sterol bearing a lactone functionality.

Animals↗

RNA interference of VCP/p97 increases Mallory body formation.

In the present report, valosin-containing protein (VCP) was present in Mallory bodies (MBs). To determine if VCP plays a role in MB formation, primary cultured hepatocytes from drug-primed mice that spontaneously form MBs in vitro were studied. The results were compared with control normal hepatocytes. Gene-specific FITC-labeled gripNA (gVCP) was added to the medium of the primary cultures to inhibit the expression of VCP. gVCP increased MB formation by 230% in drug-primed mouse hepatocytes compared with primed liver cells where no VCP oligos were added. Blocking VCP expression induced both multiple small ubiquitin (Ub) and cytokeratin (CK) aggregates to form within the cytoplasm in normal mouse hepatocytes. Inhibition of VCP expression in both drug-primed and control hepatocytes caused a decrease in proteasome chymotrypsin-like (ChT-L) activity. Overexpression of VCP was achieved by transfecting the hepatocytes with a plasmid containing green fluorescent protein (GFP)-fused VCP (pVCP-GFP). Overexpressed VCP was located in both the cytoplasm and nucleus of pVCP-GFP overexpressing drug-primed hepatocytes. VCP was also concentrated within MBs. MB formation was not decreased by the overexpression of VCP in the cells. These results indicate that VCP plays an important role in inducing MB formation, probably through its molecular chaperone function in the ubiquitin-proteasome system (UPS).

Adenosine Triphosphatases↗

The role of laminin-integrin signaling in triggering MB formation. An in vivo and in vitro study.

It is still unclear as to how hepatocytes perceive external factors and transduce the signals which initiate MB formation. To investigate this phenomenon, the model of MB formation in liver in vivo and in primary culture of hepatocytes derived from drug-primed mice was used. Control mice were fed the control diet (group 1). MBs were induced in the livers of mice fed diethyl-1, 4-dihydro-2, 4, 6-trimethyl-3, 5-pyridinedicarboxylate (DDC) for 10 weeks (group 2). The induced MBs completely disappeared after the withdrawal of DDC for 4 weeks (group 3). Newly formed MBs were numerous after DDC was refed for 1 week (group 4). Relative mRNA abundance was determined by quantitative real-time RT-PCR in the liver from the mice. The expression of integrin alpha(6) and beta(2) was significantly increased in the livers of DDC-treated (group 2) and drug refed mice (group 4), when compared with the livers from controls (group 1) and DDC-withdrawn (group 3) mice. The increased mRNA of these two integrin genes was associated with the increased expression of laminin (a ligand for integrin alpha(6)beta(1) and alpha(6)beta(4)), Icam1 (a ligand of alphaLbeta2), Src, MEKK1, and ERK1. Primary cultures of isolated DDC-primed hepatocytes (group 4 mice were withdrawn from DDC-CMZ for 4-6 weeks) produced significantly more MBs on laminin-coated coverslips compared with plastic uncoated, fibronectin-, collagen-, or fibrinogen-coated coverslips. U0126, an inhibitor of MEK1 protein, significantly reduced the phosphorylated forms of ERK1/2 and MB formation in vitro. In conclusion, the current study revealed an association between MB formation and integrin-mediated signaling in vivo. The data indicate that laminin-integrin signaling which activates ERK, triggered MB formation in vitro, and an inhibitor of the signaling cascade reduced MB formation.

Animals↗

Microarray analysis of gene expression in the liver during the urinary ethanol cycle in rats fed ethanol intragastrically at a constant rate.

The regulation of gene expression in the liver of ethanol fed rats was studied using microarray analysis. The changes in gene expression were compared between pair-fed controls and rats fed ethanol intragastrically at a constant rate for 1 month. The rats fed ethanol were sacrificed at low and high urinary ethanol levels (UAL) during the UAL cycle in order to compare the effects of high and low blood alcohol levels (BAL). The results of the microarray analysis indicated that the pattern of gene expression was very different when the controls, high UAL, and low UAL livers were compared. Many of the gene expression changes reflected nonparenchymal cell alterations such as neuroendocrine or muscle related gene expression. When the results of protein levels and gene expression were compared for individual genes, correlations were variable indicating that post-translational factors modified the effects of the changes in gene expression. This was further emphasized by the fact that activation of proteins by phosphorylation as the result of signaling kinase cascades, was not reflected in the microarray analysis results. For instance, the importance of blood alcohol levels at the time that the assays were performed, profoundly changed the gene expression, protein level, and protein phosphorylation level profiles.

Animals↗

Increased expression and function of integrins in enterocytes by endotoxin impairs epithelial restitution.

BACKGROUND & AIMS: Experimental necrotizing enterocolitis (NEC) is characterized by circulating endotoxin (lipopolysaccharide [LPS]) and impaired enterocyte migration. We hypothesized that LPS increases integrin function and cell-matrix adhesion, leading to impaired enterocyte migration in the pathogenesis of NEC. METHODS: NEC-like intestinal injury was induced in newborn rats by hypoxia/gavage feedings, and restitution was determined by assessing bromodeoxyuridine-labeled enterocytes along the crypt-villus axis. Newborn mice were injected with 5 mg/kg LPS. IEC-6 cells were treated with LPS +/- LY294002 or wortmannin, and beta 1- and alpha 3-integrins were assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunofluorescence. Beta 1-integrin function was determined by adherence of fibronectin beads to IEC-6 monolayers. Migration of IEC-6 cells into a scraped wound was measured by time-lapse microscopy. RESULTS: Newborn intestinal injury was associated with decreased intestinal restitution and increased alpha 3- and beta 1-integrin expression in the ileal mucosa, which also was observed after LPS injection. In IEC-6 cells, LPS caused an increase in the expression of alpha 3- and beta 1-integrins, a shift of beta 1-integrins from the cytoplasm to the plasma membrane and an increase in fibronectin bead adhesion during which beta 1-integrins accumulated underneath attached beads. These effects could be reversed with LY294002 or wortmannin, suggesting phosphatidylinositol-3-phosphate kinase (PI3K) dependence. The increased integrin-matrix adhesion by LPS led to an inhibition of enterocyte migration, which could be reversed by anti-beta 1-antibodies. CONCLUSIONS: Enterocyte migration is inhibited by LPS through increased expression and function of alpha 3- and beta 1-integrins. Modulation of enterocyte migration via integrins may provide novel insights into the pathogenesis of NEC, in which intestinal restitution is impaired.

Androstadienes↗

6-hydroxy-4-en-3-one sterols from the marine sponge Iotrochoto birotulata.

Five sterols with a nucleus skeleton of 6-hydroxy-4-en-3-one, namely cholesta-6beta-hydroxy-4-en-3-one (1), ergosta-6beta-hydroxy-4,24(28)-dien-3-one (2), ergosta-6alpha-hydroxy-4,24(28)-dien-3-one (3), ergosta-6alpha-hydroxy-4,22-E-dien-3-one (4), and ergosta-28-methyl-6beta-hydroxy-4,24(28)-dien-3-one (5) have been isolated from the marine sponge Iotrochoto birotulata, collected from the southern China sea. Sterols 2-4 are new compounds, and 1 has been isolated from marine organisms for the first time. The structures have been elucidated on the basis of extensive spectroscopic and chemical properties.

Animals↗

A numerical study of worker exposure to a gaseous contaminant: variations on body shape and scalar transport model.

Three-dimensional computational fluid dynamics simulations are used to investigate the distribution and level of contaminant concentrations in the true breathing zone (at the nose and mouth) when toxic airborne contaminants are released within an arm's length in front of the worker who has his back to the airflow. The effects of different body shapes on fluid flow and concentration patterns around the body in a wind tunnel were evaluated and clarified that a sharp body or a block may not be a good surrogate for the human form in consideration of occupational and environmental health studies. The comparison of the concentration field calculated with the Eulerian and Lagrangian methods revealed that the Eulerian method has a more diffusive nature than the Lagrangian method. The concentrations at different locations were also compared to determine the optimum sampling location. It was found that the concentration at the breathing zone may be significantly different from the one at the chest area. The influence of the heat flux from the body was studied at two different Reynolds numbers. Predictions indicate that the heat flux may have a significant impact on exposure especially when the convection induced by buoyancy dominates the flow.

Air Pollutants↗

Effects of beta-glucan extracted from Saccharomyces cerevisiae on humoral and cellular immunity in weaned piglets.

Twenty-four barrows were used to investigate the effects of beta-glucan on immune function in weaned piglets. Pigs (8.09 +/- 0.20 kg, 28 d of age) were fed a diet without or with supplemented beta-glucan (50 mg/kg feed). All pigs were injected with ovalbumin (OVA) on day 14 to investigate their humoral immune response. On day 28, lymphocytes were isolated from all pigs to determine the effects of beta-glucan on cellular immunity of pigs in vitro. Lymphocytes from six pigs of each group were incubated with 16 microg lipopolysaccharide (LPS) per ml culture medium, the remainder with an equivalent volume of culture medium alone. Samples were collected at 0, 3, 6, 12, 18, 24, and 48 h after LPS addition for determination of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), and interleukin-10 (IL-10). On day 31, six pigs of each group were injected with either LPS (25 microg/kg BW) or an equivalent amount of sterile saline. Blood samples were collected at 3 h after LPS injection for analysis of IL-6, TNF-alpha, and IL-10 in plasma. The results indicated that dietary beta-glucan enhanced pig antibody response to OVA only in the first week after injection. In vitro, the increases of IL-6 and TNF-alpha in culture medium were partially dampened in pigs supplemented with beta-glucan when their lymphocytes were incubated with LPS, whereas the increase of IL-10 was potentiated. In vivo, dietary beta-glucan attenuated the increase of plasma IL-6 and TNF-alpha, and enhanced the increase of plasma IL-10 when pigs were challenged with LPS. These results demonstrate that beta-glucan can improve the humoral immunity of pigs and modulate cellular immunity of pigs by mitigating the elevation of pro-inflammatory cytokines and enhancing the increase of anti-inflammatory cytokines after an immunological challenge.

Animal Feed↗

Lower plasma alpha-carboxyethyl-hydroxychroman after deuterium-labeled alpha-tocopherol supplementation suggests decreased vitamin E metabolism in smokers.

BACKGROUND: Cigarette smoking increases the fractional disappearance rates of alpha-tocopherol and is associated with increased oxidative stress, but its effects on alpha-tocopherol metabolism are unknown. OBJECTIVE: We hypothesized that smokers would have less alpha-tocopherol available and consequently lower plasma alpha-carboxyethyl-hydroxychroman (alpha-CEHC), the alpha-tocopherol metabolite produced by a cytochrome P450-mediated process. DESIGN: Smokers and nonsmokers (n = 10 per group) were supplemented with deuterium-labeled alpha-tocopheryl acetates (75 mg each d3-RRR-alpha-tocopheryl and d6-all-rac-alpha-tocopheryl acetate) from day -6 to day -1, and plasma tocopherols and CEHCs were measured (day -6 through day 17). RESULTS: After 6 d of supplementation, plasma d3- and d6-alpha-tocopherol concentrations did not differ significantly between groups. Plasma d3- and d6-alpha-CEHCs were detectable only from day -5 to day 5. Before supplementation, unlabeled alpha- and gamma-CEHCs were approximately 60% and 40% lower, respectively, in smokers than in nonsmokers (P < or = 0.05). In addition, d0-, d3-, and d6-alpha-CEHC areas under the curves were approximately 50% lower in smokers (P < 0.05), and smokers had lower maximal d3-alpha-CEHC (P = 0.004) and d6-alpha-CEHC (P = 0.0006) concentrations. Notably, 2.9-4.7 times as much alpha-CEHC was produced from all-rac-alpha-tocopherol than from RRR-alpha-tocopherol. During supplementation, smokers had about one-half (P < 0.05) the plasma total, d6-, or d3-alpha-CEHC concentrations that nonsmokers did given similar alpha-tocopherol concentrations. CONCLUSIONS: Smoking did not increase alpha-tocopherol disappearance through P450-mediated tocopherol metabolism. Therefore, the mechanism of increased alpha-tocopherol disappearance in smokers likely operates through oxidation pathways, which is consistent with alpha-tocopherol's antioxidant function. Consequently, evaluating the molecular mechanism or mechanisms responsible for tocopherol metabolism under conditions of oxidative stress and the mechanisms that regulate alpha-tocopherol status is warranted.

Adult↗

Bone mesenchymal stem cells transplanted into rabbit intervertebral discs can increase proteoglycans.

We sought to determine whether transplanted allogeneic bone mesenchymal stem cells can survive and increase the amount of proteoglycans in intervertebral discs. We used the rabbit intervertebral disc as a model, creating three groups: an uninjected control group, a group injected with saline, and a group injected with 1 x 10(5) of bone mesenchymal stem cells containing trace marker gene LacZ from young rabbits. At 1, 3, and 6 months, X-gal staining and DNA-polymerase chain reaction of the neomycin-resistant gene were used to ascertain cell location. Reverse transcription-polymerase chain reaction and enzyme-linked immunosorbant assay analysis were done to evaluate the effect on the disc matrix. Transplanted mesenchymal stem cells were located and identified in the group injected with mesenchymal stem cells, whereas we detected none in the saline and control groups. The amount of mRNA and protein of proteoglycan and collagen Type II in the mesenchymal stem cells group was increased, whereas the amount of collagen Type I did not change. We found no changes in the saline group. Our data suggest transplanted allogenic bone mesenchymal stem cells can survive and increase proteoglycan amount, supporting its potential use as a treatment of intervertebral disc degeneration.

Aggrecans↗

Pharmacokinetic and pharmacodynamic interaction between irbesartan and hydrochlorothiazide in renal hypertensive dogs.

Combined irbesartan/hydrochlorothiazide (HCTZ) formulations are often used clinically. Pharmacokinetic-pharmacodynamic (PK/PD) modeling was applied to investigate the pharmacokinetic and pharmacodynamic interaction between irbesartan and HCTZ in renal hypertensive dogs at non-steady-state and steady-state. The renal hypertensive dogs were treated with oral irbesartan alone, or HCTZ alone, or the combination of irbesartan and HCTZ for 8 days. Blood pressure and plasma concentrations were measured and pharmacokinetic-pharmacodynamic parameters were analyzed. Irbesartan showed a two-compartment model pharmacokinetic profile. The concentration-time course of irbesartan was not changed by HCTZ, but irbesartan increased the peak plasma concentration and area under the curve of HCTZ at steady-state. HCTZ had no blood pressure lowering effect at non-steady-state. Irbesartan plus HCTZ had greater blood pressure lowering action than irbesartan alone. HCTZ increased actions of irbesartan. Hysteresis loops were found between effect and plasma concentrations of irbesartan after a single dose. However, hysteresis loops disappeared at steady state with more rapid realization of maximum concentration and effects. The relationship between effects and effect-compartment concentrations of the drugs was represented by a sigmoid Emax model. The results suggest synergistic pharmacodynamic interaction between irbesartan and HCTZ in renal hypertensive dogs and some differences of pharmacokinetic-pharmacodynamic properties between irbesartan and irbesartan/HCTZ combinations at non-steady-state and steady state.

Animals↗

Induction of mucosal and systemic immune response by single-dose oral immunization with biodegradable microparticles containing DNA encoding HBsAg.

The purpose of this work was to assess the ability of plasmid DNA encoding hepatitis B virus (HBV) HBsAg encapsulated in poly(DL-lactide-co-glycolic acid) (PLGA) microparticles to induce local and systemic HBsAg-specific immunity following a single dose of oral immunization. RT-PCR analysis demonstrated prolonged transcription of plasmid DNA, consistent with the sustained expression and presentation of target antigen observed by confocal laser scanning microscopy, in gut-associated lymphocyte tissue (GALT) from mice immunized orally with plasmid DNA encapsulated into PLGA microparticles. Oral administration of PLGA-DNA microparticles induced a long-lasting and stable antigen-specific antibody response, both serum total antibody and intestinal IgA, in BALB/c mice. Mice immunized orally exhibited antigen-specific gamma interferon production and cytotoxic T lymphocyte responses in spleen and GALT after restimulation in vitro with HBsAg or tumour cells stably expressing HBsAg. In contrast, naked DNA vaccines given by intramuscular injection induced only systemic cellular and humoral responses to HBsAg, which were much lower than the responses elicited by oral DNA encapsulated in PLGA microparticles at equivalent doses. The results are encouraging with regard to obtaining good compliance and vaccination coverage with candidate plasmid DNA vaccines, especially in developing countries.

Administration, Oral↗

Living-related liver transplantation for Wilson's disease.

Orthotopic liver transplantation has been applied to the treatment of Wilson's disease (WD), living-related liver transplantation (LRLT) has also been indicated for WD with increasing frequency. Between January 2001 and November 2003, 22 LRLTs were performed on patients (19 pediatric, three adults) with WD in liver transplantation center. Two patients were transplanted because of a presentation coexistent with fulminant hepatic failure. Twenty presented with chronic advanced liver disease with (n = 9) or without (n = 11) associated neurologic manifestations. All the recipients had low serum ceruloplasmin levels with a mean value of 12.8 +/- 3.2 mg/dl before transplantation and increased to an average of 26.0 +/- 3.6 mg/dl after LRLT at the latest evaluation. The survival patients with neurologic manifestations such as tremor, dysarthia, dysphagia, dystonia and sialorrhea had improved after LRLT. This suggests that LRLT not only resolves the hepatic but also ameliorates the neurologic consequences of WD.

Adolescent↗