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Biomedical subjects

B Wang

Publications and source records attributed to B Wang.

At least 469 records · Page 26Linked to original sources

[The investigation for lysozyme conformation changes in deuteromethanol solution by FT-Raman spectrometer].

Lysozyme conformation in deuteromethanol-deuteroxide solution has been studied by FT-Raman spectrometer. By using amide I mode in the Raman spectra and curve fitting method, information of Lysozyme's secondary structure is easily detected in details, and tertiary structure changes can be suggested by relative intensity changes of some bands. It is concluded that lysozyme conformation may be changed to loose structure with incrasing deuteromethanol concentration, and it seems like that increasing loose beta-sheet content lead to the gel's appearing in the solution.

Methanol↗

[A quantitative study on secondary structure of proteins by FT-Raman spectroscopy].

In this work, proteins have been scanned by FT-Raman spectrometer. Deconvolved spectra, second derivative or fourth derivative spectra were employed to determine positions of individual components. Curve-fitting procedures were carried out to amide I band of original spectra. The broad band was resolved into several components. The resolved band components were assigned to specific protein conformations. The areas of individual components are used to estimate the percentage of the relative secondary structure. Results are in reasonable agreement with data obtained by other methods.

Animals↗

Functional anatomy of the male feline urethra: morphological and physiological correlations.

Anatomical and histological methods were combined with measurements of the urethral pressure profile (UPP) to investigate the functional aspects of the urethra in male cats. A silicone rubber catheter with two microdiaphragm pressure transducers was used to measure the UPP. Gross anatomy and ultrastructure of the urethra at each segment were examined and correlated with the pressure profile data. The preprostatic urethra was composed of three layers of smooth muscle, while distal to the prostate striated muscle became predominant. Increased baseline pressure and rapid fluctuations in pressure in the postprostatic urethra and bulbourethra resulted from the function of periurethral striated musculature. The UPP was affected by the bladder pressure, repetition of the measurement, the sensor orientation in the urethra, and the type of measurement catheter. Well controlled high fidelity measurements enabled a clear correlation to be established between the features of the UPP and the anatomy of the urethra and surrounding musculature. Additionally, observations on the ultrastructural and microscopic anatomy of the urethra extend a previous description of the pelvic urethra.

Animals↗

Radiation-induced apoptosis and limb teratogenesis in embryonic mice.

In utero irradiation of the fetus during the period of organogenesis induces a dramatic increase in malformation. However, the mechanisms underlying the teratogenesis remain to be elucidated. In the present study, the correlation between radiation-induced apoptosis and limb malformation was examined in mice. The mice were exposed to X rays in utero on day 11 of gestation during the period of organogenesis of limb buds. A marked increase in the number of apoptotic cells in the predigital regions in the forelimb buds was detected 4 h after irradiation. The preinterdigital regions of the forelimb buds did not show such an increase at the same time. Aphlangy and ectrodactyly were the main types of anomalies observed on day 19 in the limbs of the fetuses irradiated with 5 Gy. The increases in prenatal death and teratogenesis in limb digits in living fetuses were dependent on dose. The possible mechanisms involved are discussed.

Abnormalities, Radiation-Induced↗

Lewis (FUT3) genotypes in two different Chinese populations.

The allelic frequencies of the alpha (1,3/4)fucosyltransferase gene (FUT3) in two different Chinese populations (138 individuals in Shenyang and 154 in Guangzhou) were investigated using PCR-RFLP and nucleotide sequencing methods. The common alleles in the Oriental population, Le (wild type allele), le59,508 (with the mutations at nucleotide (nt) 59T-->G and nt 508G-->A) and le59,1067 (with the mutations at nt 59T-->G and nt 1067T-->A) were encountered, and also the rare alleles, le1067 (with the mutation at nt 1067T-->A) and Le59 (with the mutation at 59T-->G), were observed in these Chinese populations. In addition, the common allele in Caucasians, le202,314 (with the mutations at nt 202T-->C and nt 314C-->T), was found in the Oriental population for the first time. The allelic frequencies of the Le, Le59, le59,508, le59,1067, le202,314, and le1067, were 0.750, 0.011, 0.145, 0.054, 0.036, and 0.004 in the Shenyang population and 0.675, 0.026, 0.14, 0.123, 0.026, and 0.010 in the Guangzhou population, respectively. The presence of the alleles containing either the 59 mutation (Le59) or the 1067 mutation (le1067) suggested that the allele le59,1067 may have originated by recombination between them.

Asian People↗

Granzyme B mimics apical caspases. Description of a unified pathway for trans-activation of executioner caspase-3 and -7.

Granzyme B (GrB) is predicted to trigger apoptosis by activating preferred caspases, but the zymogens that are directly processed by the granzyme and the requirements for these interactions remain unclarified. We examined this dilemma by comparing the kinetics and pattern of GrB-mediated activation of the executioner caspase-7 in vitro and in vivo. GrB rapidly activates procaspase-7 in vitro by cleaving between the large and small subunits leaving the propeptide intact. During GrB-mediated apoptosis, the caspase-7 propeptide is removed and cleavage occurs between the subunits. Strikingly, caspase-7 is unprocessed in caspase-3-deficient MCF-7 cells exposed to GrB but is rapidly activated when the cells are solubilized. Transfection with caspase-3 restores the removal of the caspase-7 propeptide and the capacity of GrB to subsequently activate the caspase. The data suggest that GrB activates caspase-3, which then removes the propeptide of caspase-7 allowing activation by GrB. Thus GrB initiates the death pathway by processing the accessible caspase-3, and the caspase-7 propeptide regulates trans-activation of the zymogen by granzyme. As a consequence, two proteases, caspase-3 and GrB, are required to activate procaspase-7.

Apoptosis↗

Restraint stress-induced immunosuppression by inhibiting leukocyte migration and Th1 cytokine expression during the intraperitoneal infection of Listeria monocytogenes.

In this study, a murine model of Listeria monocytogenes infection was used to investigate effects of restraint stress (RST) on host defense. We observed that the L. monocytogenes infection as well as RST induced an elevation of endogenous corticosterone (CORT) levels and RST synergistically enhanced endogenous CORT levels during the listerial infection. RST suppressed the migration of leukocytes including macrophages, neutrophils, NK cells and lymphocytes into the peritoneal cavities after the intraperitoneal inoculation of L. monocytogenes. RST also suppressed the increase of the surface MHC class II antigen expression in both peritoneal macrophages and B cells during the listerial infection. Interestingly, gene expression of iNOS, MCP-1 (JE) and Th1-type cytokines including IFN-gamma and IL-12 was down-regulated but Th2-type cytokine (IL-4 and IL-6) gene expression in the PEC was rather up-regulated on day 7 after infection, indicating that Th2-type immune response is more resistant to the elevated endogenous CORT levels than Th1-type response. Treatment of mice with RU486, a glucocorticoid receptor antagonist, restored the immune responses suppressed by RST to their normal levels in the infected mice, suggesting that the RST-induced elevation of endogenous corticosterone levels is mainly responsible for the induction of the immunosuppressive events during L. monocytogenes infection.

Animals↗

Human cathepsin F. Molecular cloning, functional expression, tissue localization, and enzymatic characterization.

A cDNA for a novel human papain-like cysteine protease, designated cathepsin F, has been cloned from a lambdagt10-skeletal muscle cDNA library. The nucleotide sequence encoded a polypeptide of 302 amino acids composed of an 88-residue propeptide and a 214-residue mature protein. Protein sequence comparisons revealed 58% homology with cathepsin W; about 42-43% with cathepsins L, K, S, H, and O; and 38% with cathepsin B. Sequence comparisons of the propeptides indicated that cathepsin F and cathepsin W may form a new cathepsin subgroup. Northern blot analysis showed high expression levels in heart, skeletal muscle, brain, testis, and ovary; moderate levels in prostate, placenta, liver, and colon; and no detectable expression in peripheral leukocytes and thymus. The precursor polypeptide of human recombinant cathepsin F, produced in Pichia pastoris, was processed to its active mature form autocatalytically or by incubation with pepsin. Mature cathepsin F was highly active with comparable specific activities toward synthetic substrates as reported for cathepsin L. The protease had a broad pH optimum between 5.2 and 6.8. Similar to cathepsin L, its pH stability at cytosolic pH (7.2) was short, with a half-life of approximately 2 min. This may suggest a function in an acidic cellular compartment. Transient expression of T7-tagged cathepsin F in COS-7 cells revealed a vesicular distribution of the gene product in the juxtanuclear region of the cells. However, contrary to all known cathepsins, the open reading frame of the cathepsin F cDNA did not encode a signal sequence, thus suggesting that the protease is targeted to the lysosomal compartment via an N-terminal signal peptide-independent lysosomal targeting pathway.

Amino Acid Sequence↗

A mouse carrying genetic defect in the choice between T and B lymphocytes.

Transgenic mice with human CD3epsilon gene have been shown to exhibit early arrest of T cell development in the thymus. The present study shows that, instead of T cells, B cells are generated in the thymus of a line, tg epsilon26, of the human CD3epsilon transgenic mice. The accumulation of mature B cells in the thymus was found only in tg epsilon26 mice, not in other human CD3epsilon transgenic mouse lines or other T cell-deficient mice, including CD3-epsilon knockout mice and TCR-beta/TCR-delta double knockout mice. Hanging drop-mediated transfer into 2-deoxyguanosine-treated thymus lobes showed that lymphoid progenitor cells rather than thymus stromal cells were responsible for abnormal B cell development in tg epsilon26 thymus, and that tg epsilon26 fetal liver cells were destined to become B cells in normal thymus even in the presence of normal progenitor cells undergoing T cell development. These results indicate that lymphoid progenitor cells in tg epsilon26 mice are genetically defective in thymic choice between T cells and B cells, generating B cells even in normal thymus environment. Interestingly, tg epsilon26 thymocytes expressed GATA-3 and TCF-1, but not LEF-1 and PEBP-2alpha, among T cell-specific transcription factors that are involved in early T cell development, indicating that GATA-3 and TCF-1 expressed during thymocyte development do not necessarily determine the cell fate into T cell lineage. Thus, tg epsilon26 mice provide a novel mouse model in that lineage choice between T and B lymphocytes is genetically defective.

Animals↗

Essential and partially overlapping role of CD3gamma and CD3delta for development of alphabeta and gammadelta T lymphocytes.

CD3gamma and CD3delta are two highly related components of the T cell receptor (TCR)-CD3 complex which is essential for the assembly and signal transduction of the T cell receptor on mature T cells. In gene knockout mice deficient in either CD3delta or CD3gamma, early thymic development mediated by pre-TCR was either undisturbed or severely blocked, respectively, and small numbers of TCR-alphabeta+ T cells were detected in the periphery of both mice. gammadelta T cell development was either normal in CD3delta-/- mice or partially blocked in CD3gamma-/- mice. To examine the collective role of CD3gamma and CD3delta in the assembly and function of pre-TCR and in the development of gammadelta T cells, we generated a mouse strain with a disruption in both CD3gamma and CD3delta genes (CD3gammadelta-/-). In contrast to mice deficient in either CD3gamma or CD3delta chains, early thymic development mediated by pre-TCR is completely blocked, and TCR-alphabeta+ or TCR-gammadelta+ T cells were absent in the CD3gammadelta-/- mice. Taken together, these studies demonstrated that CD3gamma and CD3delta play an essential, yet partially overlapping, role in the development of both alphabeta and gammadelta T cell lineages.

Animals↗

[Epidemiological study of airway responsiveness on the patients with asthma].

The analysis of date on 641 nuclear families with asthma declared that the sensity and specify of the airway responsiveness with Methecholone(MTCH) Challenge Test for the clinical diagnosis of asthma is 72 percent and 70 percent. The population with airway hyperresponsiveness was 6.01 times higher risk to asthma than one of negative airway responsiveness. The OR of male and female for asthma was 4.97, 5.06, respectively, and was statistically significant (P < 0.01). Moreover, the risk of asthma was been increasing with MTCH concentration was increased (i.e. degree of airway responsiveness decreased) step by step, as well showed the lineage trend between the asthma and airway responsiveness significantly (P < 0.01). The Measurement of airway responsiveness play an important role in clinic and prevent medicine.

Adolescent↗

Amperometric glucose biosensor based on sol-gel organic-inorganic hybrid material.

A new type of sol-gel organic-inorganic hybrid material was developed and used for the production of biosensors. This material is composed of silica sol and a grafting copolymer of poly(vinyl alcohol) with 4-vinylpyridine. It prevents the cracking of conventional sol-gel-derived glasses and eliminates the swelling of the hydrogel. The optimum composition of the hybrid material was first examined, and then glucose oxidase was immobilized in this matrix to demonstrate its application. The characteristics of the biosensor were studied by cyclic voltammetry and chronoamperometry. The biosensor exhibited a series of good properties: high sensitivity (600 nA mmol-1 L-1), short response time (11 s) and remarkable long-term stability in storage (at least 5 months). In addition, the characteristics of the second-generation biosensor with the use of tetrathiafulvalene as a mediator were discussed.

Biosensing Techniques↗

Radio-sensitive murine thymoma cell line 3SB: characterization of its apoptosis-resistant variants induced by repeated X-irradiation.

3SB, a mouse thymoma cell line, is one of the most radio-sensitive cells (D0 = 0.3 Gy), and its rapid apoptosis (4 h after 5 Gy irradiation, 90% apoptosis) seems to play a decisive role in enhancing the radiosensitivity. To understand the molecular mechanisms underlying extremely high radiosensitivity and rapid apoptosis, we attempted to isolate X-ray-resistant (XR) variants from 3SBH5, a stable subclone of 3SB, by repeating exposure of the cells to 2-5 Gy X-rays. Four independent stable XR variants, R111, R223, R316 and R429, were isolated by the repeated irradiation protocols. All XR cells possessed about 3 times higher D10 values than that of their parental 3SBH5. They were also resistant to apoptosis; only 10% cells underwent apoptosis 4 h after 5 Gy irradiation. The p53 protein was induced in all the cell lines after 5 Gy X-irradiation. These variants showed a cross resistance to a chemical reagent daunorubicin (DNR) that is known to be involved in the ceramide-mediated apoptosis. DNR, as well as C2-ceramide (5 muM) induced apoptosis in parental 3SBH5 cell, but not in two XR variants, R233 and R316 cells. Present result suggests that the induction of X-ray resistance by repeated X-irradiation might be achieved, at least partly, by the enhanced resistance to the ceramide-mediated apoptosis.

Animals↗

14-3-3zeta binds a phosphorylated Raf peptide and an unphosphorylated peptide via its conserved amphipathic groove.

14-3-3 proteins bind a variety of molecules involved in signal transduction, cell cycle regulation and apoptosis. 14-3-3 binds ligands such as Raf-1 kinase and Bad by recognizing the phosphorylated consensus motif, RSXpSXP, but must bind unphosphorylated ligands, such as glycoprotein Ib and Pseudomonas aeruginosa exoenzyme S, via a different motif. Here we report the crystal structures of the zeta isoform of 14-3-3 in complex with two peptide ligands: a Raf-derived phosphopeptide (pS-Raf-259, LSQRQRSTpSTPNVHMV) and an unphosphorylated peptide derived from phage display (R18, PHCVPRDLSWLDLEANMCLP) that inhibits binding of exoenzyme S and Raf-1. The two peptides bind within a conserved amphipathic groove on the surface of 14-3-3 at overlapping but distinct sites. The phosphoserine of pS-Raf-259 engages a cluster of basic residues (Lys49, Arg56, Arg60, and Arg127), whereas R18 binds via the amphipathic sequence, WLDLE, with its two acidic groups coordinating the same basic cluster. 14-3-3 is dimeric, and its two peptide-binding grooves are arranged in an antiparallel fashion, 30 A apart. The ability of each groove to bind different peptide motifs suggests how 14-3-3 can act in signal transduction by inducing either homodimer or heterodimer formation in its target proteins.

14-3-3 Proteins↗

Identification of the spinal neural network involved in coordination of micturition in the male cat.

In these studies, we used the expression of Fos protein to identify cells within the spinal cord that regulate micturition in male cats. The immediate early gene c-fos that encodes the Fos protein can be induced rapidly and transiently in neurons by increased electrical activity. Animals were anesthetized with alpha-chloralose, and received one of four stimulus protocols: electrical stimulation of the pelvic nerve, electrical stimulation of the pudendal nerve, a period of isometric micturition (induced by ligating the proximal urethra and infusing saline into the bladder), or electrical stimulation of Barrington's nucleus. After the period of stimulation, the animals were perfused and neurons expressing Fos-like immunoreactivity (FLI) were visualized with immunocytochemical methods. Stimulation with each protocol resulted in a substantially larger number of neurons expressing FLI than in operated but unstimulated controls, which exhibited few Fos-positive neurons localized to the superficial dorsal horn. In animals undergoing isometric micturition or stimulation of Barrington's nucleus, neurons exhibiting FLI were found bilaterally in the sacral (S1-S3) spinal cord and were localized to the lateral portion of the superficial dorsal horn (laminae I and II), in the intermediolateral region (lateral laminae V-VII), and around the central canal (lamina X and medial laminae V-VII). The intermediolateral region appeared to contain two populations of cells exhibiting FLI: a group of large multipolar cells and a group of small round cells. Few Fos-immunoreactive nuclei were observed in the medial portion of the superficial dorsal horn, and FLI was not observed in ventral horn neurons. Electrical stimulation of the pudendal or pelvic nerves resulted in fewer numbers of cells exhibiting FLI, with a less widespread spatial distribution. These results identify spinal neurons that are active during the micturition cycle, and demonstrate that a behaviorally relevant stimulus (isometric micturition) generated more widespread and greater intensity of Fos expression than repetitive electrical stimulation of the component peripheral nerves.

Animals↗

Scintimammographic analysis of nonpalpable breast lesions previously identified by conventional mammography.

BACKGROUND: In randomized trials, screening mammography has led to decreased mortality from breast cancer. However, the low positive predictive value of mammography (i.e., the proportion of patients with a positive test result who actually have breast cancer) results in a large number of unnecessary biopsies. We determined whether scintimammography with technetium-99m-sestamibi is a useful supplemental diagnostic tool for women with nonpalpable breast abnormalities identified by conventional mammography. METHODS: Scintimammography was performed preoperatively on 70 women who were 31-66 years of age (mean age and median age = 51 years). These women had nonpalpable breast abnormalities identified by conventional mammography; subsequently, a needle-localization excisional biopsy of each suspicious lesion was performed. Scintimammographic images were interpreted independently by two nuclear medicine physicians who were blinded to all clinical and pathologic data, and an interobserver variation analysis was performed. RESULTS: Interobserver variation analysis of the scintimammographic findings showed an agreement for breast diagnosis of 97% and a kappa coefficient of 0.90. Comparison of scintimammographic findings and histopathologic results revealed that the sensitivity (proportion of patients with breast cancer who had a positive test result), the specificity (proportion of patients without breast cancer who had a negative test result), the positive predictive value and the negative predictive value (proportion of patients with a negative test result who actually did not have breast cancer) of scintimammography were 56% (95% confidence interval [CI] = 23%-85%), 87% (95% CI = 75%-94%), 38% (95% CI = 15%-68%), and 93% (95% CI = 82%-98%), respectively. Four of nine breast cancers were not detected by scintimammography. CONCLUSION: Because of excellent interobserver agreement, scintimammography provides an objective way of detecting primary breast carcinoma. In view of its low sensitivity and positive predictive value, however, scintimammography is not currently recommended as a screening test in patients with nonpalpable positive mammographic findings.

Adult↗