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Biomedical subjects

Lihong Shi

Publications and source records attributed to Lihong Shi.

9 recordsLinked to original sources

SARS-CoV-2 infection and vaccination elicit distinct pharyngeal mucosal B cell responses in children.

Mucosal immunity is an important correlate of protection against respiratory infections such as SARS-CoV-2. Comparing B cell responses in the upper respiratory tract following vaccination and infection may offer unique insights into mucosal immunity. Here, we characterized antigen-specific B cells in the tonsils, adenoids, and peripheral blood of children who had been infected with SARS-CoV-2 or vaccinated with SARS-CoV-2 mRNA vaccines. SARS-CoV-2-specific switched memory B cells (BSM) and germinal center B cells were found in the blood and pharyngeal lymphoid tissues after vaccination or infection. However, infection generated a higher proportion of IgA+ BSM and CXCR3+CD21+ BSM, which showed distinct spatial localization, greater clonal expansion and increased propensity for plasma cell differentiation compared to their CXCR3- counterparts, accompanied by persistent activation of innate and T follicular helper cells in the tissues. Our data provide evidence for tissue-specific B cell memory after either SARS-CoV-2 vaccination or infection, but with distinct characteristics that can influence the quality, durability, and localization of immunity.

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Electrochemiluminescent detection based on solid-phase extraction at tris(2,2'-bipyridyl)ruthenium(II)-modified ceramic carbon electrode.

A sensitive electrochemiluminescent detection scheme by solid-phase extraction at Ru(bpy)3(2+)-modified ceramic carbon electrodes (CCEs) was developed. The as-prepared Ru(bpy)3(2+)-modified CCEs show much better long-term stability than other Nafion-based Ru(bpy)3(2+)-modified electrodes and enjoy the inherent advantages of CCEs. The log-log calibration plot for dioxopromethazine is linear from 1.0 x 10(-9) to 1.0 x 10(-4) mol L(-1) using the new detection scheme. The detection limit is 6.6 x 10(-10) mol L(-1) at a signal-to-noise ratio of 3. The new scheme improves the sensitivity by approximately 3 orders of magnitude, which is the most sensitive Ru(bpy)3(2+) ECL method. The scheme allows the detection of dioxopromethazine in a urine sample within 3 min. Since Ru(bpy)3(2+) ECL is a powerful technique for determination of numerous amine-containing substances, the new detection scheme holds great promise in measurement of free concentrations, investigation of protein-drug interactions and DNA-drug interactions, pharmaceutical analysis, and so on.

Journal Article↗

Static shear modulus of electrorheological fluids.

We report measurements of the static shear modulus of electrorheological (ER) fluids consisting of water-wetted silica microspheres in silicone oil. A shear-annealing method, using creep-recovery (CR) cycles under an external electric field, is used to enhance ER properties of the fluid. The shear-annealing method enables the silica spheres in the ER fluid to form better aligned and denser column microstructures. A stable state with elastic shear deformation is obtained after a sufficient number of CR cycles, with an optimal combination of stress duration and shear strength. Static shear modulus is obtained by measuring the elastic deformations at different shear stresses for an electric field frequency from 10 to 1000 Hz. A water-bridge model is proposed to explain the enhanced shear modulus.

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Influence of composition on the photorefractive centers in pure LiNbO3 at low light intensity.

The influence of composition on the photorefractive effect in pure LiNbO3 crystals at low light intensity was investigated. The experimental results indicate that different defects dominate the photorefractive centers of pure LiNbO3 with various compositions. Bipolarons are considered to be responsible for the enhanced photovoltaic field in reduced near-stoichiometric LiNbO3, and their bulk photovoltaic constant kappa is estimated to be approximately 6.95 x 10(-32) m3/V. Q polarons (composed of two bipolarons) are introduced to explain the photorefractive effect of congruent LiNbO3 at both low and high light intensities.

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Application of ceramic carbon materials for solid-phase extraction of organic compounds.

Ceramic carbon materials were developed as new sorbents for solid-phase extraction of organic compounds using chlorpromazine as a representative. The macroporosity and heterogeneity of ceramic carbon materials allow extracting a large amount of chlorpromazine over a short time. Thus, the highly sensitive and selective determination of chlorpromazine in urine sample was achieved by differential pulse voltammograms after only 1-min extraction. The total analysis time was less than 3 min. In comparison with other electrochemical and electrochemiluminescent methods following 1-min extraction, the proposed method improved sensitivity by about 2 and 1 order of magnitude, respectively. The fast extraction, diversity, and conductivity of ceramic carbon materials make them promising sorbents for various solid-phase extractions, such as solid-phase microextraction, thin-film microextraction, and electrochemically controlled solid-phase extraction. The preliminary applications of ceramic carbon materials in chromatography were also studied.

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Structural and kinetic analysis of the substrate specificity of human fibroblast activation protein alpha.

Fibroblast activation protein alpha (FAPalpha) is highly expressed in epithelial cancers and has been implicated in extracellular matrix remodeling, tumor growth, and metastasis. We present the first high resolution structure for the apoenzyme as well as kinetic data toward small dipeptide substrates. FAPalpha exhibits a dipeptidyl peptidase IV (DPPIV)-like fold, featuring an alpha/beta-hydrolase domain and an eight-bladed beta-propeller domain. Known DPPIV dipeptides are cleaved by FAPalpha with an approximately 100-fold decrease in catalytic efficiency compared with DPPIV. Moreover, FAPalpha, but not DPPIV, possesses endopeptidase activity toward N-terminal benzyloxycarbonyl (Z)-blocked peptides. Comparison of the crystal structures of FAPalpha and DPPIV revealed one major difference in the vicinity of the Glu motif (Glu(203)-Glu(204) for FAPalpha; Glu(205)-Glu(206) for DPPIV) within the active site of the enzyme. Ala(657) in FAPalpha, instead of Asp(663) as in DP-PIV, reduces the acidity in this pocket, and this change could explain the lower affinity for N-terminal amines by FAPalpha. This hypothesis was tested by kinetic analysis of the mutant FAPalpha/A657D, which shows on average an approximately 60-fold increase in the catalytic efficiency, as measured by k(cat)/K(m), for the cleavage of dipeptide substrates. Furthermore, the catalytic efficiency of the mutant is reduced by approximately 350-fold for cleavage of Z-Gly-Pro-7-amino-4-methylcoumarin. Our data provide a clear understanding of the molecular determinants responsible for the substrate specificity and endopeptidase activity of FAPalpha.

Amino Acid Motifs↗

N-linked glycosylation of dipeptidyl peptidase IV (CD26): effects on enzyme activity, homodimer formation, and adenosine deaminase binding.

The type II transmembrane serine protease dipeptidyl peptidase IV (DPPIV), also known as CD26 or adenosine deaminase binding protein, is a major regulator of various physiological processes, including immune, inflammatory, nervous, and endocrine functions. It has been generally accepted that glycosylation of DPPIV and of other transmembrane dipeptidyl peptidases is a prerequisite for enzyme activity and correct protein folding. Crystallographic studies on DPPIV reveal clear N-linked glycosylation of nine Asn residues in DPPIV. However, the importance of each glycosylation site on physiologically relevant reactions such as dipeptide cleavage, dimer formation, and adenosine deaminase (ADA) binding remains obscure. Individual Asn-->Ala point mutants were introduced at the nine glycosylation sites in the extracellular domain of DPPIV (residues 39-766). Crystallographic and biochemical data demonstrate that N-linked glycosylation of DPPIV does not contribute significantly to its peptidase activity. The kinetic parameters of dipeptidyl peptidase cleavage of wild-type DPPIV and the N-glycosylation site mutants were determined by using Ala-Pro-AFC and Gly-Pro-pNA as substrates and varied by <50%. DPPIV is active as a homodimer. Size-exclusion chromatographic analysis showed that the glycosylation site mutants do not affect dimerization. ADA binds to the highly glycosylated beta-propeller domain of DPPIV, but the impact of glycosylation on binding had not previously been determined. Our studies indicate that glycosylation of DPPIV is not required for ADA binding. Taken together, these data indicate that in contrast to the generally accepted view, glycosylation of DPPIV is not a prerequisite for catalysis, dimerization, or ADA binding.

Adenosine Deaminase↗

Shear-enhanced yield stress in electrorheological fluids.

We report the enhancement of yield stress in electrorheological (ER) fluids by a shear-annealing method, using creep-recovery (CR) cycles under an external electric field. The enhancement depends on the duration and the strength of the applied shear stress, as well as on the number of the CR cycles. The shear-annealing method enables the particles in the ER fluid to form microstructures with an increased yield stress, manifest as better aligned and denser columns. For a sufficiently large number of CR cycles, with an optimal combination of stress duration and shear strength, a stable state can be obtained whereby shear deformation becomes elastic.

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