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Biomedical subjects

X Chen

Publications and source records attributed to X Chen.

At least 487 records · Page 27Linked to original sources
PubMed2000

Co-expression of interleukin-2 and green fluorescent protein reporter in mycobacteria: in vivo application for monitoring antimycobacterial immunity.

Recombinant Mycobacterium bovis bacillus Calmette-Guérin expressing green fluorescent protein (rBCG-GFP), driven by the mycobacterial heat shock protein 70 (HSP 70) promoter from an autonomously replicating plasmid, was genetically engineered to co-express mouse interleukin-2 (IL-2) by introduction of an independent HSP 60 promoter. To monitor host antimycobacterial immunity, C57BL/6 mice were intravenously infected with IL-2 expressing and non-expressing GFP rBCGs. Both rBCGs were clearly imaged and easily quantified with ultraviolet microscopy of tissue sections and whole organ suspensions. Enhanced mycobacterial clearance from the spleens of mice infected with the rBCG-IL-2/GFP strain was apparent by both diminished bacterial counts and spleen weights during the first 6 weeks post-infection relative to rBCG-GFP. T helper type 1 (TH1) cytokine production and proliferative response to BCG restimulation was also elevated from in vitro splenocyte cultures taken from the rBCG-IL-2/GFP-infected group. Taken together, these results suggest that IL-2 expression from rBCG augmented host protective immunity to mycobacterial infection via an enhanced TH1 immune response. Mycobacterial expression vectors that allow simultaneous but independent production of reporter proteins and bioactive substances provide an ideal means for monitoring the in vivo fate of recombinant mycobacteria.

Animals↗
PubMed2000

Long form leptin receptor mRNA expression in the brain, pituitary, and other tissues in the pig.

Much effort has focused recently on understanding the role of leptin, the obese gene product secreted by adipocytes, in regulating growth and reproduction in rodents, humans and domestic animals. We previously demonstrated that leptin inhibited feed intake and stimulated growth hormone (GH) and luteinizing hormone (LH) secretion in the pig. This study was conducted to determine the location of long form leptin receptor (Ob-Rl) mRNA in various tissues of the pig. The leptin receptor has several splice variants in the human and mouse, but Ob-Rl is the major form capable of signal transduction. The Ob-Rl is expressed primarily in the hypothalamus of the human and rodents, but has been located in other tissues as well. In the present study, a partial porcine Ob-Rl cDNA, cloned in our laboratory and specific to the intracellular domain, was used to evaluate the Ob-Rl mRNA expression by RT-PCR in the brain and other tissues in three 105 d-old prepuberal gilts and in a 50 d-old fetus. In 105 d-old gilts, Ob-Rl mRNA was expressed in the hypothalamus, cerebral cortex, amygdala, thalamus, cerebellum, area postrema and anterior pituitary. In addition, Ob-Rl mRNA was expressed in ovary, uterine body, liver, kidney, pancreas, adrenal gland, heart, spleen, lung, intestine, bone marrow, muscle and adipose tissue. However, expression was absent in the thyroid, thymus, superior vena cava, aorta, spinal cord, uterine horn and oviduct. In the 50 d-old fetus, Ob-Rl mRNA was expressed in brain, intestine, muscle, fat, heart, liver and umbilical cord. These results support the idea that leptin might play a role in regulating numerous physiological functions.

Animals↗
PubMed2000

Contrast harmonic color Doppler left ventriculography: machine-interpreted left ventricular ejection fraction compared with equilibrium-gated radionuclide ventriculography.

BACKGROUND: Multi-gated acquisition (equilibrium-gated radionuclide ventriculography) (MUGA) is considered the gold standard for measuring left ventricular ejection fraction (LVEF) because it is accurate, machine interpreted, and reproducible. Echocardiographic LVEF measurements are subject to variability in image acquisition and interpretation and to the limitations of 2-dimensional (2D) versus 3-dimensional imaging. GOAL: The shortcomings of traditional echocardiography may be addressed by combining multiplane 2D harmonic imaging, echocardiographic contrast, color Doppler ultrasonography, and digital image processing to create a new imaging modality: contrast harmonic color Doppler left ventriculography. METHODS: We compared the accuracy of a new method for measuring LVEF that allows for machine interpretation and uses contrast-enhanced intermittent harmonic color Doppler ultrasonography (CHCD). Quantitative LVEF measurements by hand-traced harmonic 2D echocardiography, contrast-enhanced harmonic 2D echocardiography, CHCD, and machine-interpreted CHCD were compared with MUGA in 35 patients. RESULTS: Contrast-enhanced intermittent harmonic color Doppler provided images with vivid endocardial definition in all patients, but hand-traced harmonic 2D echocardiography and contrast-enhanced harmonic 2D echocardiography had inadequate images in 9% of patients. The MUGA LVEF range was 0. 09 to 0.70. All echocardiographic methods showed excellent correlation with the MUGA LVEF (R (2) > 0.96), but the CHCD method had the best limits of agreement. CONCLUSIONS: Contrast-enhanced intermittent harmonic color Doppler LVEF correlates with MUGA at least as well as traditional noncontrasted echocardiography, but it provides diagnostic images in a greater proportion of patients. The CHCD images have vivid endocardial delineation and can be machine interpreted.

Adult↗
PubMed2000

Novel action of pituitary adenylate cyclase-activating polypeptide. Stimulation of extracellular acidification in rat pituitary GH4C1 cells.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a member of the vasoactive intestinal peptide/secretin family. Using microphysiometry, we have found that PACAP acutely (1 min) increased the extracellular acidification rate (ECAR) in GH4C1 cells approximately 40% above basal in a concentration-dependent manner. ECAR, maximally induced by PACAP, can be increased further by thyrotropin-releasing hormone (TRH), indicating that the signalling pathways for these two neuropeptides are not identical. In studies on the mechanism of PACAP-enhanced ECAR, we found that maximum stimulation of the cAMP/PKA pathway by treatment with FSK, or the PKC pathway with PMA, did not inhibit the ECAR response to PACAP. The PKC inhibitor calphostin C and the MAP kinase inhibitor PD98059 had no effect on the ECAR response to PACAP. Furthermore, PACAP induced little or no change in cytosolic Ca(2+) ([Ca(2+)](i)), while TRH induced a large increase in [Ca(2+)](i). However, the tyrosine kinase inhibitor genistein completely blocked PACAP-induced ECAR, suggesting involvement of tyrosine kinase(s). We conclude that PACAP causes an increase in ECAR in GH4C1 rat pituitary cells, which is not dependent on the PKA, PKC, MAP kinase or Ca(2+) signalling pathways, but does require tyrosine kinase activity.

Animals↗
PubMed2000

Characterization of the 16S rRNA gene V2 region and the rrn operons of Gardnerella vaginalis.

Ribosomal RNA (rRNA) gene polymorphism was apparent when Gardnerella vaginalis DNA restriction profiles were hybridized with nonradioactively labeled total rRNA isolated from this bacterium. In contrast, use of a polymerase chain reaction (PCR)-based 16S rRNA gene V2 region resulted in a 118-bp V2-PCR amplicon that was specific and common in all 30 tested G. vaginalis isolates. In addition to providing a G. vaginalis-specific fingerprint, when the V2-PCR amplicon along with total rRNA were utilized as probes, a partial rRNA gene restriction map could be constructed. G. vaginalis contains two rrn operons with an EcoRI fragment of 1.6 kb common to both.

DNA Fingerprinting↗
PubMed2000

Stereoselective pharmacokinetics of propafenone and its major metabolites in healthy Chinese volunteers.

The stereoselective pharmacokinetics of propafenone (PPF) and its active metabolite 5-hydroxypropafenone (5-OHP) as well as their glucuronide and sulfate conjugates have been investigated, in order to clarify the relationship between metabolism and stereoselective disposition of PPF in humans. After oral administration of 300 mg racemic PPF hydrochloride to 10 healthy Chinese subjects, the areas under the plasma concentration-time curves (AUCs) for (S)-PPF were significantly higher (S/R ratio, 1.50+/-0.17) and the apparent oral clearance significantly lower (S/R ratio, 0.68+/-0.07) than those parameters for (R)-PPF. In contrast, the AUCs of PPF glucuronide (PPF-G) were lower for (S)-PPF-G than for the (R)-enantiomer (S/R ratio, 0.83+/-0.12). The partial clearance of (S)-PPF by glucuronidation pathway was lower than that of (R)-PPF and the enantiomeric ratio was 0.62+/-0.04. The t(max) values of PPF glucuronide diastereoisomers showed no statistically significant differences between each other, but were much shorter than the corresponding values of the parent drug, implying that glucuronidation may be the 'first-choice' pathway in presystemic metabolism of PPF. Glucuronidation of 5-OHP favored the (S)-enantiomer, whereas the sulfation showed a large preference for the (R)-enantiomer. After beta-glucuronidase hydrolysis, no significant differences were observed in AUCs between 5-OHP enantiomers (including unconjugated and conjugated 5-OHP). The results suggest that the significant difference in disposition between PPF enantiomers may be, at least in part attributed to stereoselective metabolism in the glucuronidation pathway.

Adult↗
PubMed2000

Dipeptide seryl-histidine and related oligopeptides cleave DNA, protein, and a carboxyl ester.

The amino acids histidine (His) and serine (Ser), or amino acids similar to Ser, function together as key catalytic amino acids in the active sites of such diverse enzymes as the serine- and thiol-proteases, lipases, and esterases. Ser and His are also conserved in the intein-extein junctions of the phylogenetically widespread self-splicing proteins and at the N- and C-termini of the homing endonucleases spliced from them. Here we show that the dipeptide seryl-histidine (Ser-His) and related oligopeptides can themselves cleave DNA, protein, and the ester p-nitrophenyl acetate (p-NPA) over wide ranges of pH and temperature. Denaturing polyacrylamide gel electrophoresis (PAGE) of 5'-end labeled DNA samples incubated with Ser-His reveals a pattern of two bands per nucleotide position, consistent with the generation of both 3'-hydroxyl and 3'-phosphate DNA cleavage fragments, as would be expected of phosphodiester hydrolysis by Ser-His. To the best of our knowledge, Ser-His is the shortest peptide ever reported to show cleavage activity with multiple categories of natural substrates. The amenability of the dipeptide to variation through addition of amino acid residues, either internally or to the C-terminus while retaining its multiple cleavage activities, combined with its reactivity over wide ranges of pH and temperature, demonstrates the evolutionary capacity of the Ser/His dyad and evokes many questions about possible roles it may have played in molecular evolution and its potential role as a core for selection of oligopeptides with enhanced cleavage activities and target specificity.

Amino Acid Sequence↗
PubMed2000

Orientation relationships of martensite variants determined by electron backscatter diffraction

Orientation relationships between the parent phase and 2H type martensite, and between different variants in a plate group of martensite in the CuAlNi shape memory alloy were determined by electron backscatter diffraction. The result reveals clearly that the basal planes of different correspondence variants of the 2H martensite originate from different 110P planes of the parent phase, and the [010]2H axis of each correspondence variant originates from one of the <001>P axes of the parent phase. Our result reveals also the typical relationships of four habit-variants A, B, C and D in a martensite plate group, and that the habit-variants A and C (and equivalenty B and D) are twin-related by an 1212H mirror plane, and the habit-variants A and D (and B and C) are twin-related by an 1012H mirror plane. Usually, variants A and C (and B and D) form spear morphology and variants A and D (and B and C) form fork morphology.

Journal Article↗
PubMed2000

ER stress induces cleavage of membrane-bound ATF6 by the same proteases that process SREBPs.

ATF6 is a membrane-bound transcription factor that activates genes in the endoplasmic reticulum (ER) stress response. When unfolded proteins accumulate in the ER, ATF6 is cleaved to release its cytoplasmic domain, which enters the nucleus. Here, we show that ATF6 is processed by Site-1 protease (S1P) and Site-2 protease (S2P), the enzymes that process SREBPs in response to cholesterol deprivation. ATF6 processing was blocked completely in cells lacking S2P and partially in cells lacking S1P. ATF6 processing required the RxxL and asparagine/proline motifs, known requirements for S1P and S2P processing, respectively. Cells lacking S2P failed to induce GRP78, an ATF6 target, in response to ER stress. ATF6 processing did not require SCAP, which is essential for SREBP processing. We conclude that S1P and S2P are required for the ER stress response as well as for lipid synthesis.

Activating Transcription Factor 6↗
PubMed2000

Analysis of neurotoxin 3-N-oxalyl-L-2,3-diaminopropionic acid and its alpha-isomer in Lathyrus sativus by high-performance liquid chromatography with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) derivatization.

A method was developed for the quantitative determination of the neurotoxic nonprotein amino acid, 3-N-oxalyl-L-2,3-diaminopropionic acid (beta-ODAP), and its nontoxic alpha-isomer, 2-N-oxalyl-L-2, 3-diaminopropionic acid (alpha-ODAP), in the plant samples of Lathyrus sativus after derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) by reversed-phase high-performance liquid chromatography (HPLC). Hippuric acid was used as an internal standard. A linear response was recorded in the concentration rang 0.32-32 nmol with r > 0.999. The RP HPLC detection limit for both isomers is 1.8 ng. According to different experimental needs, a ternary gradient system can be used to determine toxin and other nonprotein amino acids. The RP HPLC method and a colorimetric method were compared for measuring ODAP.

Amino Acids, Diamino↗
PubMed2000

Eradication of murine mammary adenocarcinoma through HSVtk expression directed by the glucose-starvation inducible grp78 promoter.

Gene therapy strategies employing the HSVtk/ganciclovir (GCV) suicide gene offer promising approaches towards the treatment of metastatic breast cancer. These include bystander effects on non-transduced tumor cells, lower systemic toxicity, and the possibility of inducing immunity against the tumor. Previously we have demonstrated the ability of the grp78 stress-inducible promoter to stimulate expression of reporter genes within the tumor microenvironment. However, experimental evidence demonstrating the ability of this promoter to activate therapeutic agents within the breast cancer environment causing tumor eradication is needed prior to clinical trials. In this report, we test the efficacy of the grp78 promoter in a retroviral system to drive the expression of the HSVtk suicide gene in a murine mammary adenocarcinoma cell line (TSA) in syngeneic, immune-competent hosts. Our results show that under glucose-starvation conditions in vitro, the expression of HSVtk and GCV induced cell death are enhanced in tumor cells in which the HSVtk gene is driven by the internal grp78 promoter compared to cells in which the Moloney murine leukemia virus LTR drives HSVtk. In in vivo studies, in tumors in which the HSVtk gene is driven by the grp78 promoter, GCV treatment causes complete tumor eradication, whereas tumors persist when the HSVtk gene is driven by the retroviral LTR. Our study suggests that the grp78 promoter may be useful to enhance the effectivity of therapeutic agents within a breast tumor. In addition, it is shown that immune memory is induced in syngeneic, immune-competent hosts. This new retroviral vector might therefore be useful for breast cancer gene therapy.

Adenocarcinoma↗
PubMed2000

Sociable and prosocial dimensions of social competence in Chinese children: common and unique contributions to social, academic, and psychological adjustment.

A sample of 6th-grade children (N = 470), initially aged 12 years, in Shanghai, People's Republic of China, participated in this longitudinal study. Data on children's sociability and prosocial orientation were collected from peer assessments. Information on indexes of adjustment was obtained from multiple sources. The results indicated that sociable and prosocial functions were 2 overlapping, but distinct, dimensions of social competence and had common as well as unique contributions to the prediction of adjustment in specific areas. Whereas prosocial orientation was mainly predictive of social and school adjustment and externalizing problems, sociability uniquely contributed to the prediction of internalizing problems and emotional adjustment. Finally, prosocial orientation and sociability might moderate the development of adjustment difficulties.

Achievement↗
PubMed2000

English language use as a risk factor for smoking initiation among Hispanic and Asian American adolescents: evidence for mediation by tobacco-related beliefs and social norms.

Acculturation increases the risk of smoking among Hispanic and Asian American adolescents, but the underlying mechanisms are not understood. This study examined associations between English language use and smoking among 4,167 Hispanic and 2,836 Asian American adolescents in California. Potential mediators were assessed, including access to cigarettes, perceived consequences, friends' smoking, cigarette offers, refusal self-efficacy, and prevalence estimates of peer smoking. English language use was associated with increased risk of lifetime smoking in both groups. This association became nonsignificant after access, perceived consequences, friends' smoking, and offers were controlled for. The acculturation process (as indicated by English language use) may be associated with smoking-related psychosocial variables, which may lead to an increased risk of experimentation with smoking.

Adolescent↗
PubMed2000

Parental warmth, control, and indulgence and their relations to adjustment in Chinese children: a longitudinal study.

A sample of children, initially 12 years old, in the People's Republic of China participated in this 2-year longitudinal study. Data on parental warmth, control, and indulgence were collected from children's self-reports. Information concerning social, academic, and psychological adjustment was obtained from multiple sources. The results indicated that parenting styles might be a function of child gender and change with age. Regression analyses revealed that parenting styles of fathers and mothers predicted different outcomes. Whereas maternal warmth had significant contributions to the prediction of emotional adjustment, paternal warmth significantly predicted later social and school achievement. It was also found that paternal, but not maternal, indulgence significantly predicted children's adjustment difficulties. The contributions of the parenting variables might be moderated by the child's initial conditions.

Adaptation, Psychological↗
PubMed2000

Interactions within the yeast t-SNARE Sso1p that control SNARE complex assembly.

In the eukaryotic secretory and endocytic pathways, transport vesicles shuttle cargo among intracellular organelles and to and from the plasma membrane. Cargo delivery entails fusion of the transport vesicle with its target, a process thought to be mediated by membrane bridging SNARE protein complexes. Temporal and spatial control of intracellular trafficking depends in part on regulating the assembly of these complexes. In vitro, SNARE assembly is inhibited by the closed conformation adopted by the syntaxin family of SNAREs. To visualize this closed conformation directly, the X-ray crystal structure of a yeast syntaxin, Sso1p, has been determined and refined to 2.1 A resolution. Mutants designed to destabilize the closed conformation exhibit accelerated rates of SNARE assembly. Our results provide insight into the mechanism of SNARE assembly and its intramolecular and intermolecular regulation.

Amino Acid Sequence↗
PubMed2000

Effects of tetrandrine on calcium transport, protein fluorescences and membrane fluidity of sarcoplasmic reticulum.

To understand whether the molecular mechanism of Tetrandrine (Tet)'s pharmacological effects is concerned with sarcoplasmic reticulum calcium transport so as to be involved in myocardial contractility, we observed the effects of Tet on calcium transport and membrane structure of rabbit skeletal muscle sarcoplasmic reticulum vesicles (SR) and rat cardiac sarcoplasmic reticulum vesicles (CSR). Calcium uptake was monitored with a dual-wavelength spectrophotometer. Protein conformation and fluorescence polarization were measured by fluospectrophotometric method and membrane lipids labelled with fluorescence probes for SR, respectively. 128 micromol l(-1) Tet reduced the initial rate of calcium uptake to 59% of control 6 min after reaction. Tet un-competitively inhibited SR Ca(2+), Mg(2+)-ATPase activity, causing the stoichiometric ratio of SR Ca(2+)/ATP to decrease to 1.43 from 2.0 of control. Inhibitory rates on SR Ca(2+),Mg(2+)-ATPase by Tet were reduced from 60% in the absence of phosphate to 50% in the presence of phosphate and reduced from 92% in 1 mmol l(-1) ATP to 60% in 5 mmol l(-1) ATP. Tet markedly reduced SR intrinsic protein fluorescence, while it slightly decreased the thiol(SH)-modified protein fluorescence of SR labelled with N-(3-pyrene)-maleimide. Tet slightly increased fluorescence polarization in the middle and deep layers of SR membrane lipids labelled with 7- or 12-(9-anthroyloxy) stearic acid (AS) probes, whereas it did not change that of SR labelled with 1, 6-diphenyl-1,3,5-hexatrine (DPH). These results revealed that prevention of SR calcium uptake by Tet was due to inhibition of the SR calcium pump Ca(2+),Mg(2+)-ATPase, changes in spatial conformation of the pumps protein molecules and a decrease in the extent of motion of membrane lipid molecules, thus altering the regulation of [Ca(2+)](i) and myocardial contractility.

Adenosine Triphosphate↗
PubMed2000

The high-sensitivity determination of protein concentrations by the enhancement of Rayleigh light scattering of Arsenazo-DBN.

A new Rayleigh light scattering (RLS) assay of protein is presented in this paper. At the optimum pH 4.10, the weak RLS of Arsenazo-DBN can be greatly enhanced by the addition of proteins due to the interaction between protein and Arsenazo-DBN. Based on this, the reactions of Arsenazo-DBN and proteins, including bovine serum albumin, human serum album, gamma-globulin, egg albumin, lysozyme and trypsin, were studied. A new quantitative determination method for proteins has been developed. The linear range for human serum albumin, for example, is 0.085-34.62 micrograms mL-1 with a detection limit of 44.8 ng mL-1. Besides high sensitivity, the method is characterized by good reproducibility, rapidity of reaction, good stability, and few interfering substances. The determination of the proteins in human serum and urine samples by this method give results very close to those obtained using Coomassie Brilliant Blue G-250 colorimetry, with relative standard deviations of 0.7-2.5%.

Arsenazo III↗