Search PubMed⌕ Search

Biomedical subjects

Z Pan

Publications and source records attributed to Z Pan.

At least 19 recordsLinked to original sources

Engineering of Bacillus subtilis for enhanced total synthesis of folic acid.

We investigated whether the yield of the B vitamin folic acid could be elevated in Bacillus subtilis. Strategies for increasing the folic acid yield were investigated by employing computer-aided flux analysis and mutation. Controlling the activity of the enzyme pyruvate kinase by placing it under inducible control was one strategy devised to elevate yield while insuring that a rapid growth rate results. Other single mutation strategies included amplifying the expression of the genes in the folate operon and overexpressing the Escherichia coli aroH gene, which encodes 2-dehydro-3-deoxyphosphoheptonate aldolase. The latter could conceivably elevate the abundance of the folic acid precursor, para-aminobenzoic acid. Strains that combined two or more mutations were also constructed. Overall, a strain possessing inducible pyruvate kinase, overexpressed aroH, and increased transcription and translation of genes from the folic operon exhibited the best yield. The yield was eightfold higher than that displayed by the parent B. subtilis 168 strain.

Bacillus subtilis↗

Non-specific tolerance induced by staphylococcal enterotoxin B in treating high risk corneal transplantation in rats.

AIMS: To explore the role of staphylococcal enterotoxin B (SEB) in treating high risk corneal keratoplasty in rats. METHODS: Rat corneal high risk transplantation rejection models were set up using Fisher 344 and Lewis rats. The experimental rats were injected intraperitoneally with 0.2 ml SEB at different concentrations before keratoplasty. The rejection indexes of the allograft were recorded and the lymphocyte infiltration in the allograft and the percentage of the lymphocyte subpopulation in the lymphatic organs were also examined. Lymphocyte proliferation ability and the concentration of IL-2 and IL-10 in the serum were also evaluated. RESULTS: Compared with the control group, SEB prolonged the survival time of the allograft significantly from 7 to 12 days. It could also reduce CD4(+) and CD8(+) lymphocyte infiltration in the allograft and minimise the percentage of CD4(+) and CD8(+) lymphocytes in the lymphatic organs. The lymphocyte proliferation ability was also weakened. However, the percentage of CD4(+) NK T lymphocytes in the lymphatic organs was raised. The serum concentration of IL-10 was higher but IL-2 was lower in the SEB treated groups. CONCLUSIONS: SEB prolonged the survival time of the allograft in high risk rat corneal allo-transplantation, which may be caused by T cell deletion and acquisition of non-specific tolerance.

Animals↗

Surface-enhanced Raman spectroscopy using silver-coated porous glass-ceramic substrates.

Surface-enhanced Raman scattering (SERS) has been studied using a silver-coated porous glass-ceramic material as a new type of substrate. The porous glass-ceramic is in the CaO-TiO2-P2O5 system prepared by controlled crystallization and subsequent chemical leaching of the dense glass-ceramic, leaving a solid skeleton with pores ranging in size from 50 nm to submicrometer. Silver was coated on the surface of the porous glass-ceramic by radio frequency (RF) sputtering or e-beam evaporation in vacuum. SERS spectra of excellent quality were obtained from several dyes and carboxylic acid molecules, including rhodamine 6G, crystal violet, isonicotinic acid, and benzoic acid, using this new substrate. This new substrate showed a good compatibility with these molecules. The porous glass ceramic with a nanometer-structured surface accommodated both test molecules and silver film. The absorbed molecules were therefore better interfaced with silver for surface-enhanced Raman scattering.

Benzoic Acid↗

SEB combined with IL-1ra could prolong the survival of the rat allografts in high-risk corneal transplantation.

PURPOSE: To determine whether the superantigen Staphylococcal enterotoxin B (SEB) combined with interleukin-1 receptor antagonist (IL-1ra) prolong allograft survival better than individual agents in high-risk corneal transplantation in a rat model. METHODS: Fisher 344 donor corneas were transplanted into Lewis recipients. High-risk transplantation meant that the transplants were sutured into the recipient beds with corneal neovascularization induced by placing three interrupted sutures in the host cornea. All of the recipients were divided in blinded fashion into four groups. Group I was injected with saline buffer. Group II was injected intraperitoneally with 0.2 mL SEB (75 microg/kg) at 4-day intervals on three occasions before transplantation. Group III was injected with 0.1 mL IL-1ra (1 mg/mL) subconjunctivally from the first day after transplantation for 2 weeks. Group IV received both SEB and IL-1ra. All transplants were evaluated for signs of rejection for 4 weeks after surgery. Ten days after transplantation, two recipients in each group were sacrificed for histopathological and immunological evaluation. RESULTS: The mean survival time of the allografts in the control group was 5.89 +/- 0.79 days; in SEB group, 10.70 +/- 2.52 days; in IL-1ra group, 8.25 +/- 0.71 days; in the SEB and IL-1ra group, 17.36 +/- 2.39 days. CD4+ and CD8+ lymphocyte infiltration into the allografts and the percentage of the lymphocytes in the spleen and mandibular lymphatic nodes was significantly decreased among the treated groups with dampened lymphocyte reactivity. The SEB plus IL-1ra combination group showed the strongest inhibition. CONCLUSION: SEB and IL-1ra are most effective in combination to treat high-risk corneal transplants.

Animals↗

A convenient and rapid sample repositioning approach for atomic force microscopy.

A novel repositioning approach is described for repeated observations of a specimen at a close proximal location in the atomic force microscope. The approach is similar to keystone architecture, whereby the repositioning is achieved by forming a male structured base for the specimen, and a corresponding female counterpart as the frame. For the combination of an acrylic acid frame and a metal base, 90% translation shifts are less than 10 microm, and almost all angular disorientations are within +3 degrees to -3 degrees . Nanometre-scale surface features can be relocated easily and reliably even after 40 imaging-removal-imaging cycles, dipping the specimen in solutions or heating up to 500 degrees C.

Microscopy, Atomic Force↗

Enterovirus replication in valvular tissue from patients with chronic rheumatic heart disease.

AIMS: To investigate the involvement of enterovirus infection in chronic, rheumatic heart disease. METHODS AND RESULTS: Formalin-fixed, paraffin-embedded, surgical samples of valve tissue were examined for the presence of enteroviral RNA and virus capsid protein VP1 by in situ hybridization and immunostaining. Of 53 cases, 33 were patients with chronic rheumatic heart disease and 20 had Marfan's syndrome or degenerative valve disease. Enterovirus RNA was detected in 8 (24.2%) of 33 patients with chronic rheumatic heart disease by in situ hybridization using strand-specific oligonucleotide probes, complementary to conserved sequences in enterovirus genomic (positive strand) RNA. The replication template (negative strand) RNA also was found in seven of these eight cases. The viral capsid protein VP1 was detected in 16 (48.5%) of 33 patients with chronic rheumatic heart disease by immunohistochemistry and correlated with viral RNA detection. Virus was localized generally to valvular tissue. Neither viral RNA nor capsid protein VP1 were found in valvular tissue from any of the 20 comparison cases. CONCLUSIONS: This is the first demonstration of detection and localization of both enterovirus RNA and capsid protein in chronic rheumatic heart disease. The presence of negative strand RNA and VP1 indicates enteroviral RNA replication and protein synthesis and suggests an aetiological role of enterovirus in the pathogenesis of chronic rheumatic heart disease.

Adult↗

RyR3 amplifies RyR1-mediated Ca(2+)-induced Ca(2+) release in neonatal mammalian skeletal muscle.

The neonatal mammalian skeletal muscle contains both type 1 and type 3 ryanodine receptors (RyR1 and RyR3) located in the sarcoplasmic reticulum membrane. An allosteric interaction between RyR1 and dihydropyridine receptors located in the plasma membrane mediates voltage-induced Ca(2+) release (VICR) from the sarcoplasmic reticulum. RyR3, which disappears in adult muscle, is not involved in VICR, and the role of the transiently expressed RyR3 remains elusive. Here we demonstrate that RyR1 participates in both VICR and Ca(2+)-induced Ca(2+) release (CICR) and that RyR3 amplifies RyR1-mediated CICR in neonatal skeletal muscle. Confocal measurements of intracellular Ca(2+) in primary cultured mouse skeletal myotubes reveal active sites of Ca(2+) release caused by peripheral coupling between dihydropyridine receptors and RyR1. In myotubes lacking RyR3, the peripheral VICR component is unaffected, and RyR1s alone are able to support inward CICR propagation in most cells at an average speed of approximately 190 microm/s. With the co-presence of RyR1 and RyR3 in wild-type cells, unmitigated radial CICR propagates at 2,440 microm/s. Because neonatal skeletal muscle lacks a well developed transverse tubule system, the RyR3 reinforcement of CICR seems to ensure a robust, uniform, and synchronous activation of Ca(2+) release throughout the cell body. Such functional interplay between RyR1 and RyR3 can serve important roles in Ca(2+) signaling of cell differentiation and muscle contraction.

Animals↗

MinK-related peptide 1: A beta subunit for the HCN ion channel subunit family enhances expression and speeds activation.

The HCN family of ion channel subunits underlies the currents I(f) in heart and I(h) and I(q) in the nervous system. In the present study, we demonstrate that minK-related peptide 1 (MiRP1) is a beta subunit for the HCN family. As such, it enhances protein and current expression as well as accelerating the kinetics of activation. Because MiRP1 also functions as a beta subunit for the cardiac delayed rectifier I(Kr), these results suggest that this peptide may have the unique role of regulating both the inward and outward channels that underlie cardiac pacemaker activity. The full text of this article is available at http://www.circresaha.org.

Animals↗

Synergistic movements of Ca(2+) and Bax in cells undergoing apoptosis.

Apoptosis is a physiological counterbalance to mitosis and plays important roles in tissue development and homeostasis. Cytosolic Ca(2+) has been implicated as a proapoptotic second messenger involved in both triggering apoptosis and regulating cell death-specific enzymes. A critical early event in apoptosis is associated with the redistribution of Bax from cytosol to mitochondria and endoplasmic reticulum (ER) membranes; however, the molecular mechanism of Bax translocation and its relationship to Ca(2+) is largely unknown. Here we provide functional evidence for a synergistic interaction between the movements of intracellular Ca(2+) and cytosolic Bax in the induction of apoptosis. Overexpression of Bax in cultured cells causes a loss of ER Ca(2+) content. Depletion of ER Ca(2+) through activation of the ryanodine receptor enhances the participation of Bax into the mitochondrial membrane. Neither Bax translocation nor Bax-induced apoptosis is affected by buffering of cytosolic Ca(2+) with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, suggesting that depletion of ER Ca(2+) rather than elevation of cytosolic Ca(2+) is the signal for cell apoptosis. This dynamic interplay of Ca(2+) and Bax movements may serve as an amplifying factor in the initial signaling steps of apoptosis.

Animals↗

Regulation of KChIP2 potassium channel beta subunit gene expression underlies the gradient of transient outward current in canine and human ventricle.

Expression of four members of the KChIP family of potassium channel beta subunits was examined in canine heart. Only one member of the gene family, KChIP2, was expressed in heart. There was a steep gradient of KChIP2 mRNA expression across the canine ventricular free wall. KChIP2 mRNA was 25-fold more abundant in the epicardium than in the endocardium, and this gradient paralleled the gradient in transient outward current (Ito) expression. In contrast, Kv4.3 potassium channel alpha subunit mRNA was expressed at equal levels across the ventricular wall. There was no difference in the pharmacological sensitivity of epicardial and endocardial Ito channels to flecainide, suggesting that the current is produced by the same channel in the two tissues. A similar gradient of KChIP2 expression was found across the ventricular wall of human heart, but not rat heart. It is concluded that transcriptional regulation of the KChIP2 beta subunit gene, rather than the Kv4.3 [alpha] subunit gene, is the primary determinant regulating the transmural gradient of Ito expression in the ventricular free wall of canine and human heart.

Animals↗

Alternative splicing of KCNQ2 potassium channel transcripts contributes to the functional diversity of M-currents.

The region of alternative splicing in the KCNQ2 potassium channel gene was determined by RNase protection analysis of KCNQ2 mRNA transcripts. Systematic analysis of KCNQ2 alternative splice variant expression in rat superior cervical ganglia revealed multiple variant isoforms. One class of KCNQ2 splice variants, those that contained exon 15a, was found to have significantly different kinetics to those of the other isoforms. These transcripts encoded channel subunits that, when co-expressed with the KCNQ3 subunit, activated and deactivated approximately 2.5 times more slowly than other isoforms. Deletion of exon 15a in these isoforms produced a reversion to the faster kinetics. Comparison of the kinetic properties of the cloned channel splice variants with those of the native M-current suggests that alternative splicing of the KCNQ2 gene may contribute to the variation in M-current kinetics seen in vivo.

Alternative Splicing↗

Simultaneous quantitative UV spectrophotometric determination of multicomponents of amino acids using linear neural network.

Simultaneous determination of multicomponents of six amino acids with a novel chemometric technique-a linear neural network (LNN) algorithm is reported in this study. Based on the data correlation coefficient and standard deviation method, 17 representative wavelength points are selected from the original UV spectral data (343 points) as the original input patterns for LNN to build a neural network model. The results obtained only by iterating 15 times is satisfying, with a correlation coefficient of 0.999 and a relative small standard deviation.

Algorithms↗

Computerized planimetry versus clinical assessment for the measurement of cervical ectopia.

OBJECTIVE: As part of a study to assess the role of cervical ectopia in the acquisition of cervical infections, we determined the reliability of cervical ectopia measurements made by computer planimetry and by clinical (visual) assessment. STUDY DESIGN: We conducted pelvic examinations of 1004 women seeking contraceptive services at two health centers in Baltimore. After application of acetic acid, clinicians estimated the relative area of ectopia by visual inspection and took cervical photographs. Two independent raters measured the absolute and relative areas of ectopia from the digitized images by means of an analytic software program. Agreement levels between raters, between multiple readings by the same rater, and between the two measurement methods were quantified by means of the intraclass correlation coefficient and weighted kappa. RESULTS: Intrarater agreement was excellent for computer planimetry measurements of the absolute (intraclass correlation coefficient, 0.97) and relative (intraclass correlation coefficient, 0.89) areas of ectopia. Interrater agreement was also high for computer planimetry measurements of the absolute (intraclass correlation coefficient, 0.83) and relative (intraclass correlation coefficient, 0.85) areas of ectopia. Agreement levels were moderate between clinician assessment and computer planimetry measurements of the relative area of ectopia (kappa = 0.48), but agreement was better when clinical assessment was limited to observations by a single, experienced clinician. CONCLUSION: Measurement of cervical ectopia by computer planimetry was highly reliable and appears appropriate for assessment of the role of ectopia in the acquisition of cervical infections. Clinical assessment of cervical ectopia may be used when computer planimetry is not available.

Cervix Uteri↗

[Combustion characteristics of municipal solid waste in fluidized bed].

For the experimental investigation on the combustion characteristics of Municipal Solid Waste(MSW) in Fluidized Bed, a lab scale Fluidized Bed facility was constructed. Single and mixed municipal solid waste burning in Fluidized Bed showed that dry waste can be burnt quickly at the bed temperature of 500 degrees C, and furnace temperature increased about 30-50 degrees C. Many kinds of combustion runs were conducted in this Fluidized Bed combustion facility. The parameters examined were air flow rate(from 5.5 m3/h to 7.5 m3/h), form of fuels(scrap or whole), moisture content and so on. Concentration of CO2, CO, SO2 and NOx in the flue gas were monitored and recorded every 5 seconds. The temperatures along the reactor were recorded every 10 seconds.

Hot Temperature↗

[Acidity and acid buffering capacity of aerosols during sand-dust storm weather in Beijing].

In the spring of 2000, there were 12 sand-dust storms in Beijing. 2 sand-dust storms were experienced in time and mass concentrations, elementary concentrations acidity and acidic buffering capacity of TSP(Total Suspended Particulate, < 100 microns) and PM10(Inhalable particulate, < 10 microns) were sampled and analyzed. Results showed that pollution level of aerosols was extremely high. However, the acidity of aerosols was relatively low and the aerosols had very strong acid buffering capacity for acidification. Therefore, the aerosols brought about by the sand-dust storms could avoid the occurrence of acidic precipitation to some extent.

Acids↗

[Efficacy of topical cyclosporine A on keratoplasty rejection in rats].

OBJECTIVE: To evaluate the efficacy of topical cyclosporine A (CsA) on a penetrating keratoplasty rejection model in rats. METHODS: Lou rats received orthotopic corneal allografts from inbred F344 donors. The rats were treated with one drop of topical solution four times daily for 30 days. Sixty rats were divided into one control (the matrix of CsA solution) and five treatment groups (0.5%, 1.0%, 2.0% CsA, glucocorticoid, glucocorticoid and 1.0% CsA). Rejection index (RI) and mean survival time (MST) were calculated, and histopathological and immunohistopathological examination were performed for each group. RESULTS: The graft MST was postponed by topical CsA treatment. There was significant difference between each treatment group and the control group (t greater-than-or-equal 2.28, P < 0.0l), especially 2% CsA, steroid and combined 1% CsA and steroid groups, the combined group being the most effective. The RI was lower in all the groups with topical CsA than that in the control group (P < 0.01). The effect of 2% CsA was the same as that of the steroid, while 1% CsA combined steroid was the most effective. The histopathological findings confirmed that the topical application of CsA reduced the inflammatory cells infiltrating the graft stroma and the neovascularization. Immunohistopathological analysis demonstrated that there were fewer CD(+)(11a) T cells and macrophages in the infiltration and intercellular adhesion molecule (ICAM)-1 positive cells in keratocytes and endothelium of grafts in the treatment groups than in the control group. CONCLUSIONS: Topical CsA can inhibit the keratoplasty rejection episodes in the rat and afford better results when combined with topical steroid.

Animals↗

[An experimental study on subconjunctival interleukin-1 receptor antagonist for promotion of corneal transplant survival].

OBJECTIVE: To determine whether the subconjunctival application of interleukin-1 receptor antagonist (IL-1ra) can prolong the corneal graft survival in the rat model of orthotopic penetrating keratoplasty. METHODS: For all experiments, F344 corneas were transplanted into LOU (major histocompatibility-disparate) eyes. Experimental groups received subconjunctival injection of 50, 100 and 200 microg IL-1ra respectively, and the control group received the same volume of 0.9% normal saline instead for consecutive 2 weeks. All transplants were evaluated for 4 weeks after surgery for signs of rejection. RESULTS: The mean survival time (MST) of the grafts of the experimental groups was increased significantly (t = 0.00, P < 0.01) in comparison with the control group. The MST of the IL-1ra 200 microg group was increased significantly than that of the IL-1ra 50 microg group (t = 0.00, P < 0.01). Furthermore, the IL-1ra-treated grafts had significantly less corneal inflammation, infiltration, lower levels of opacity, edema, neovascularization and rejection index compared with the control group. CONCLUSIONS: Subconjunctival treatment of IL-1ra has a significantly positive effect on promoting corneal allograft survival. And its effect is dosage-dependent.

Animals↗