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Biomedical subjects

H Yuan

Publications and source records attributed to H Yuan.

At least 19 recordsLinked to original sources
PubMed2002

Preparation of solid lipid nanoparticles with clobetasol propionate by a novel solvent diffusion method in aqueous system and physicochemical characterization.

Solid lipid nanoparticles (SLN) are a colloidal carrier system for controlled drug delivery. Monostearin SLN were prepared by a novel solvent diffusion method in an acidic aqueous system in order to improve the recovery of the method. The lipophilic model drug clobetasol propionate was incorporated to study the recovery of nanoparticles, entrapment efficacy, zeta potential (charge) and drug delivery characterization. The drug and monostearin were dissolved in acetone and ethanol at 50 degrees C in water bath, the resultant organic solution was poured into an acidic aqueous (pH 1.10) containing 1% polyvinyl alcohol (PVA) under mechanical agitation at room temperature. The drug loaded SLN was quickly produced with an aggregation state and easily separated by centrifugation. The recovery of nanoparticles was markedly increased compared to using a usual aqueous (pH 5.73) containing the same concentration of PVA. After burst drug release at the first 3 h, a distinctly prolonged release over a monitored period of 4 days was observed and nearly 6% drug was released in each day. Further, a novel preparation method and the optimized separation parameters in the present research for SLN were established. These results also demonstrate the principle suitability of SLN as a prolonged release formulation for lipophilic drugs.

Algorithms↗
PubMed2001

Application of solid-phase microextraction in the determination of diazepam binding to human serum albumin.

In this paper, protein-drug interactions were studied by solid-phase microextraction (SPME) using diazepam binding to human serum albumin as a model system. Since drug compounds are normally polar and nonvolatile by nature, direct SPME is used in this work. The SPME extraction is an equilibrium process among the concentrations of the analyte partitioned onto the SPME fiber, free and bound drug in the solution. A calibration curve was first constructed by employing the amount of the analytes partitioned on the fiber versus the free analyte concentration in the solution in the absence of protein. In method I, the extraction was performed in the protein solution with known diazepam concentration. In method II, diazepam was first loaded onto the fiber by extracting in solution with known diazepam concentration. This fiber was subsequently transferred into the protein solution for desorption. The amount of the analyte left on the fiber was analyzed after the system reached equilibrium. The free drug concentration was then obtained from the calibration curve for both methods. The Scatchard plot was finally employed to obtain the number of binding sites and the equilibrium binding constants. Since only a very small amount of the protein solution is required (150 microL for each extraction), method II is very useful for circumstances where the protein amount is very limited. The direct measurement method proposed in this paper does not need a GC response factor, which significantly decreases the experimental error. The only measurement needed is the area count change (ratio) of the fiber injections before and after the protein was introduced into the solution. The difference between the direct measurement method for method I and method II is discussed. The result illustrated that the SPME direct measurement method provided both theoretical accuracy and simplicity in such applications.

Calibration↗
PubMed2001

Mutations in the novel protocadherin PCDH15 cause Usher syndrome type 1F.

We have determined the molecular basis for Usher syndrome type 1F (USH1F) in two families segregating for this type of syndromic deafness. By fluorescence in situ hybridization, we placed the human homolog of the mouse protocadherin Pcdh15 in the linkage interval defined by the USH1F locus. We determined the genomic structure of this novel protocadherin, and found a single-base deletion in exon 10 in one USH1F family and a nonsense mutation in exon 2 in the second. Consistent with the phenotypes observed in these families, we demonstrated expression of PCDH15 in the retina and cochlea by RT-PCR and immunohistochemistry. This report shows that protocadherins are essential for maintenance of normal retinal and cochlear function.

Adult↗
PubMed2001

Protective role of ATP-sensitive potassium channels in hypoxia-induced generalized seizure.

Adenosine triphosphate (ATP)-sensitive potassium (K(ATP)) channels are activated by various metabolic stresses, including hypoxia. The substantia nigra pars reticulata (SNr), the area with the highest expression of K(ATP) channels in the brain, plays a pivotal role in the control of seizures. Mutant mice lacking the Kir6.2 subunit of K(ATP) channels [knockout (KO) mice] were susceptible to generalized seizures after brief hypoxia. In normal mice, SNr neuron activity was inactivated during hypoxia by the opening of the postsynaptic K(ATP) channels, whereas in KO mice, the activity of these neurons was enhanced. K(ATP) channels exert a depressant effect on SNr neuronal activity during hypoxia and may be involved in the nigral protection mechanism against generalized seizures.

Adenosine Triphosphate↗
PubMed2001

Bone induction by porous glass ceramic made from Bioglass (45S5).

Porous glass ceramic, which was prepared from Bioglass powder (45S5, U.S. Biomaterials) by foaming with diluted H(2)O(2) solution and sintering at 1000 degrees C for 2 h, was implanted as cylinders (5 mm in diameter and 6 mm in length) in thigh muscles of dogs for 3 months. Histological observation was made on thin un-decalcified sections. Bone formation was histologically found in pores of all implants (X16) retrieved from 16 dogs. The bone tissue was also identified with backscattered scanning electron microscopy observation (BSE) and energy dispersive X-ray microanalysis (EDX). This is the first report of bone induction in soft tissues of animals by glass ceramic that has long been recognized as a bioactive (osteoconductive) biomaterial. The present results justify the impetus to investigate the osteoinductivity of calcium phosphate-based biomaterials, to study the mechanism of bone induction (osteoinduction) by calcium phosphate-based biomaterials, to develop osteoinductive calcium phosphate-based biomaterials, and to examine the relation between osteoinduction and osteoconduction.

Animals↗
PubMed2001

Purification, refolding of hybrid hIFNgamma-kringle 5 expressed in Escherichia coli.

The DNA sequence coding for plasminogen kringle 5 (pK5), an inhibitor of angiogenesis, was fused with that coding for interferon gamma and over-produced in the form of inactive inclusion bodies in E. coli. The amount of fusion protein was about 40% of total protein produced. The fusion protein contained in the inclusion bodies was solubilized in 8 M urea and purified by anion-exchange chromatography. We employed the orthogonal experimental design L16(4(5)) (5 factors, 4 levels, 16 experiments) procedure for researching the influence of denaturant, aggregation suppressor L-arginine, NaCl, pH, and glycine on the refolding procedure. Our results suggest that the presence of appropriate L-arginine, NaCl, and denaturant in the refolding buffer inhibits the aggregation of the fusion protein and increases the yield of renatured protein with biological activity. The refolded fusion protein, gammaIFN/pk5, has in vitro anti-endothelial cell proliferation activity.

Angiogenesis Inhibitors↗
PubMed2001

Material-dependent bone induction by calcium phosphate ceramics: a 2.5-year study in dog.

Bone induction by different calcium phosphate biomaterials has been reported previously. With regard to (1) whether the induced bone would disappear with time due to the absence of mechanical stresses and (2) whether this heterotopically formed bone would give rise to uncontrolled growth, a long-time investigation of porous hydroxyapatite ceramic (HA), porous biphasic calcium phosphate ceramic (TCP/HA, BCP), porous alpha-tricalcium phosphate ceramic (alpha-TCP) and porous beta-tricalcium phosphate ceramic (beta-TCP) was performed in dorsal muscles of dog, for 2.5 years. Histological observation, backscattered scanning electron microscopy observation and histomorphometric analysis were made on thin un-decalcified sections of retrieved samples. Normal compact bone with bone marrow was found in all HA implants (n = 4) and in all BCP implants (n = 4), 48 +/- 4% pore area was filled with bone in HA implants and 41 +/- 2% in BCP implants. Bone-like tissue, which was a mineralised bone matrix with osteocytes but lacked osteoblasts and bone marrow, was found in all beta-TCP implants (n = 4) and in one of the four alpha-TCP implants. Both normal bone and bone-like tissues were confined inside the pores of the implants. The results show that calcium phosphate ceramics are osteoinductive in muscles of dogs. Although the quality and quantity varied among different ceramics, the induced bone in both HA and BCP ceramics did neither disappear nor grow uncontrollably during the period as long as 2.5 years.

Animals↗
PubMed2001

Synthesis and bioactivity of 2,4-diacyl analogues of paclitaxel.

The 2,4-diacyl paclitaxel analogues 8a-8r were prepared from paclitaxel by acylation of 4-deacetyl-2-debenzoylpaclitaxel 1,2-carbonate (3) followed either by hydrolysis of the carbonate and acylation or by direct treatment of the carbonate with an aryllithium. Some of the resulting derivatives showed significantly improved tubulin assembly activity and cytotoxicity as compared with paclitaxel; in some cases this improvement was especially significant for paclitaxel-resistant cell lines.

Antineoplastic Agents, Phytogenic↗
PubMed2001

Low-density lipoprotein receptor-related protein-5 binds to Axin and regulates the canonical Wnt signaling pathway.

To understand how the Wnt coreceptor LRP-5 is involved in transducing the canonical Wnt signals, we identified Axin as a protein that interacts with the intracellular domain of LRP-5. LRP-5, when expressed in fibroblast cells, showed no effect on the canonical Wnt signaling pathway by itself, but acted synergistically with Wnt. In contrast, LRP-5 mutants lacking the extracellular domain functioned as constitutively active forms that bind Axin and that induce LEF-1 activation by destabilizing Axin and stabilizing beta-catenin. Addition of Wnt caused the translocation of Axin to the membrane and enhanced the interaction between Axin and LRP-5. In addition, the LRP-5 sequences involved in interactions with Axin are required for LEF-1 activation. Thus, we conclude that the binding of Axin to LRP-5 is an important part of the Wnt signal transduction pathway.

3T3 Cells↗
PubMed2001

Biological sample analysis with immunoaffinity solid-phase microextraction.

A theophylline antiserum was covalently immobilized on the surface of a fused silica fiber, modified with 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde, and used as a selective and sensitive extraction medium for the immunoaffinity solid-phase microextraction (SPME) determination of theophylline in serum samples. The specificity of the immunoaffinity SPME fiber was first investigated using a fixed concentration of [3H]theophylline together with various amounts of interference, possessing no cross-reactivity with the theophylline antibody. No significant non-specific binding was observed. The reproducibility of the fiber preparation and the immunoaffinity SPME analysis was also investigated, resulting in a relative standard deviation of 6.1% for five analyses of the same fiber. The antigen-antibody binding isotherm was obtained by analyzing theophylline standards of various concentrations (0.1-5 ng mL(-1)) until saturation values were reached. Initial binding of theophylline was linear with a r2 = 0.968. The cross-reactivity of the theophylline immunoaffinity SPME fiber for the structural analog caffeine was investigated by adding various amounts of caffeine in the presence of theophylline at a saturation concentration and produced a low cross-reactivity value of 0.1%. Finally. spiked serum samples (10 and 50 ng mL(-1)) were successfully analyzed with an excellent correlation with the standard binding isotherm, thus confirming the performance of the immunoaffinity SPME coating for improved bioanalysis.

Animals↗
PubMed2001

Aminopeptidase A: a nephritogenic target antigen of nephrotoxic serum.

BACKGROUND: We investigated potential targets of antibody-mediated glomerular injury induced with a noncomplement binding fraction of sheep anti-rat nephrotoxic serum (NTS). This model is characterized by severe complement- and leukocyte-independent proteinuria within 24 hours of NTS injection into rats. METHODS: NTS-reactive glomerular cell and matrix proteins were identified by immunoprecipitation, Western blot analysis, protein sequencing, cDNA library screening, and enzyme-linked immunosorbent assay. Proteinuria was measured in rats injected with NTS from which reactivity against type IV collagen had been removed by immunoadsorption, and antibodies were eluted from the glomeruli of proteinuric rats that had been injected with unabsorbed NTS. Having identified aminopeptidase A (APA) as a major target of NTS, we studied the effect of NTS and anti-APA on mouse glomerular epithelial cells in culture. RESULTS: NTS identified several podocyte and matrix proteins; however, APA was the only cell surface protein reactive with antibodies eluted from the glomeruli of rats injected with NTS. Although the eluate also contained reactivity to the noncollagenous domains of alpha1 and alpha3 chains of type IV collagen, immunodepletion of these antibodies did not diminish the ability of NTS to cause proteinuria. We also documented the surface expression of APA on mouse glomerular epithelial cells in culture, and found that NTS and specific anti-APA antibodies induce a time- and temperature-dependent redistribution of the antigen. CONCLUSIONS: APA, a type II integral membrane metallopeptidase, is a major target of NTS in vivo and is known to be present on the surface of podocytes. NTS-induced proteinuria is independent of reactivity to known nephritogenic matrix proteins. These findings, in combination with previous studies showing that monoclonal anti-APA antibodies induce severe proteinuria in mice, suggest that anti-APA antibodies are responsible for complement-independent proteinuria in this model.

Aminopeptidases↗
PubMed2001

Alkaloids and flavonoids from peanut skins.

Separation of the water-soluble fraction of peanut skins led to the isolation of 8 flavonoids and two novel indole alkaloids. Two new flavonoid glycosides have been identified as isorhamnetin 3-O-[2-O-beta-glucopyranosyl-6-O-alpha-rhamnopyranosyl]- beta-glucopyranoside and 3',5,7-trihydroxyisoflavone-4'-methoxy-3'-O-beta-glucopyranoside. Two alkaloids are 2-methoxyl-3-(3-indolyl)-propionic acid and 2-hydroxyl-3-[3-(1-N-methyl)-indolyl]-propionic acid. These isolated flavonoids were evaluated for their free radical scavenging activity and protein glycation inhibitory effects.

Alkaloids↗
PubMed2001

Immunization with a pentameric L1 fusion protein protects against papillomavirus infection.

The prophylactic papillomavirus vaccines currently in clinical trials are composed of viral L1 capsid protein that is synthesized in eukaryotic expression systems and purified in the form of virus-like particles (VLPs). To evaluate whether VLPs are necessary for effective vaccination, we expressed the L1 protein as a glutathione S-transferase (GST) fusion protein in Escherichia coli and assayed its immunogenic activity in an established canine oral papillomavirus (COPV) model that previously validated the efficacy of VLP vaccines. The GST-COPV L1 fusion protein formed pentamers, but these capsomere-like structures did not assemble into VLPs. Despite the lack of VLP formation, the GST-COPV L1 protein retained its native conformation as determined by reactivity with conformation-specific anti-COPV antibodies. Most importantly, the GST-COPV L1 pentamers completely protected dogs from high-dose viral infection of their oral mucosa. L1 fusion proteins expressed in bacteria represent an economical alternative to VLPs as a human papillomavirus vaccine.

Animals↗
PubMed2001

Inhibition of host transcription by vesicular stomatitis virus involves a novel mechanism that is independent of phosphorylation of TATA-binding protein (TBP) or association of TBP with TBP-associated factor subunits.

The matrix (M) protein of vesicular stomatitis virus (VSV) is a potent inhibitor in vivo of transcription by all three host RNA polymerases (RNAP). In the case of host RNA polymerase II (RNAPII), the inhibition is due to lack of activity of the TATA-binding protein (TBP), which is a subunit of the basal transcription factor TFIID. Despite the potency of M protein-induced inhibition in vivo, experiments presented here show that M protein cannot directly inactivate TFIID in vitro. Addition of M protein to nuclear extracts from uninfected cells did not inhibit transcription activity, indicating that the inhibition is indirect and is mediated through host factors. The host factors that are known to regulate TBP activity include phosphorylation by host kinases and association with different TBP-associated factor (TAF) subunits. However, TBP in VSV-infected cells was found to be assembled normally with its TAF subunits, as shown by ion exchange high-pressure liquid chromatography and sedimentation velocity analysis. A normal pattern of phosphorylation of TBP in VSV-infected cells was also observed by pH gradient gel electrophoresis. Collectively, these data indicate that M protein inactivates TBP activity in RNAPII-dependent transcription by a novel mechanism, since the known mechanisms for regulating TBP activity cannot account for the inhibition.

DNA-Binding Proteins↗
PubMed2001

[A permanent implantable pulsatile impeller heart with a specially devised needle bearing].

The author's implantable pulsatile impeller pump can assist the circulation of the calves for several months. The termination of the experiments was related to wear of the mechanical bearing, which resulted in vibration of the rotor and pump failure. It seemed as if the experiments could have lasted indefinitely if the bearing had not failed. To solve the problem of bearing wear, a specially devised rolling bearing was investigated and applied to the impeller pump. Its service life is more than 10 years due to the wear-proof roller made of ultra-highmolecular weight polythene. This newly devised impeller heart promises to have long-term and permanent applications. Compared with the magnetic bearing, the rolling bearing has the advantages of simpler construction and control, no consumption of energy, and better reliability.

Animals↗