Characteristics of Allergic Pulmonary Inflammation in CXCR3Knockout Mice Sensitized and Challenged with House Dust Mite Protein

<div><p>Chemokine C-X-C motif receptor 3 (CXCR3) is a chemokine receptor that is mainly expressed by activated T lymphocytes. T cells play important roles in allergic pulmonary inflammation, which is a hallmark of asthma and elicits the localized accumulation of activated T cells in the lung. In China, a marked increase in the incidence rate of chronic allergic pulmonary inflammation has made it a major public health threat. In the present study, we investigated the role of CXCR3 and its ligands in airway inflammation induced by house dust mite protein (HDMP) in a CXCR3 knockout (CXCR3KO) asthma mouse model. Pathological manifestations in the lung, cell counts and bronchoalveolar lavage fluid (BALF) classifications were studied using hematoxylin and eosin (H&E) staining. The levels of IL-4 and IFN-γ in the BALF and splenocyte supernatants were measured using ELISA. CD4⁺ and CD8⁺ T cells in the lung and spleen were analyzed by flow cytometry. RT-PCR was applied to measure the mRNA transcript levels of monokines induced by IFN-γ(CXCL9) and IFN-γ inducible protein 10(CXCL10). The total cell counts, eosinophil counts, and IL-4 levels in the BALF and cultured splenocyte supernatants were significantly increased, while the levels of IFN-γ were reduced in the HDMP groups(<i>P</i><0.01). Changes in the total cell counts, eosinophil counts, and lymphocyte counts, as well as the total protein levels in the BALF, the levels of IL-4 in splenocyte supernatants, and the pathological manifestations in the lung, were all greater in CXCR3KO mice than in C57BL/6 wild-type mice. Furthermore, the expression levels of CXCL9 and CXCL10 mRNA transcripts in the lungs of CXCR3KO mice were lower than those in C57BL/6 wild-type mice (<i>P</i><0.05). CXCR3 and its ligands (i.e., CXCL9 and CXCL10) may play anti-inflammatory roles in this animal model. Promoting the expression of CXCR3 and its ligands may represent a novel therapeutic approach for preventing and curing asthma.</p></div

Levels of IL-4 and IFN-γ in Mouse Splenocyte Culture Supernatants.

<p>WTC: wild-type control group; KOC: CXCR3KO control group; WTP: wild-type HDMP test group; KOP: CXCR3KO HDMP test group. The levels of IL-4 were significantly increased in splenocyte culture supernatant derived from sensitized mice, and the levels of IFN-γ were decreased. Furthermore, the level of IL-4 was significantly different between the KOP and WTP groups; WTC: n = 4; WTP: n = 6; KOC: n = 6; KOP: n = 5; *<i>P</i><0.01, HDMP vs. control; #<i>P</i><0.01, CXCR3KO vs. wild-type.</p

Flow Cytometric Assessment of CD4⁺ and CD8⁺ T Cell Frequencies in the Lung.

<p>WTC: wild-type control group; KOC: CXCR3KO control group; WTP: wild-type HDMP test group; KOP: CXCR3KO HDMP test group. Flow cytometry was used to detect T cell subsets in the lung. Top panel, representative histogram showing the presence of CD4⁺ T cells and CD8⁺ T cells in lung tissue. The data presented one of four animals in 2 independent experiments. Bottom panel, pooled data showing the percentages of CD4⁺ T cells and CD8⁺ T cells in lung tissue; n = 4; *<i>P</i><0.05 and **<i>P</i><0.01, HDMP vs. control; #<i>P</i><0.01, CXCR3KO vs. wild-type.</p

The Proportions of CD4⁺ and CD8⁺ T Cells in the Spleen, as Determined by Flow Cytometry.

<p>A, B, C, D represent WTC, KOC, WTP and KOP group respectively. WTC: wild-type control group; KOC: CXCR3KO control group; WTP: wild-type HDMP test group; KOP: CXCR3KO HDMP test group. Top panel, representative histogram showing the presence of CD4⁺ T cells and CD8⁺ T cells in the spleen. The data presented one of four animals in 2 independent experiments. Bottom panel, pooled data showing the percentages of CD4⁺ T cells and CD8⁺ T cells in the spleen; n = 4; *<i>P</i><0.05.</p

Relative mRNA Transcript Levels of CXCL9 and CXCL10 in Lung Tissue Samples.

<p>WTC/WC: wild-type control group; KOC/KC: CXCR3KO control group; WTP/WP: wild-type HDMP test group; KOP/KP: CXCR3KO HDMP test group. The RT-PCR products were electrophoresed on an agarose gel (Fig 5A-5C). Band intensities in the resulting images were quantified using Bio-Rad Quantity One software. β-actin was used as an internal control, and the ratio of the 2 bands was used to represent the relative gene expression value (Fig 5D and 5E); n = 6 mice per group; *<i>P</i><0.05 and **<i>P</i><0.01, HDMP vs. control; #<i>P</i><0.05 and ##<i>P</i><0.01, CXCR3KO vs. wild-type.</p

Lung Histopathology (H&E staining).

<p>A, WTC group (200×); B, KOC group (200×); C, WTP group (200×); D, KOP group (200×). Mice were sensitized and challenged with HDMP. Lung sections were stained with 1% H&E reagent. Histopathological changes were observed using a light microscope, as described in the Materials and Methods section. A-D, Representative H&E images; n = 6 animals per group.</p

Multicolored pH-Tunable and Activatable Fluorescence Nanoplatform Responsive to Physiologic pH Stimuli

Tunable, ultra-pH responsive fluorescent nanoparticles with multichromatic emissions are highly valuable in a variety of biological studies, such as endocytic trafficking, endosome/lysosome maturation, and pH regulation in subcellular organelles. Small differences (e.g., <1 pH unit) and yet finely regulated physiological pH inside different endocytic compartments present a huge challenge to the design of such a system. Herein, we report a general strategy to produce pH-tunable, highly activatable multicolored fluorescent nanoparticles using commonly available pH-insensitive dyes with emission wavelengths from green to near IR range. The primary driving force of fluorescence activation between the ON (unimer) and OFF (micelle) states is the pH-induced micellization. Among three possible photochemical mechanisms, homo Förster resonance energy transfer (homoFRET)-enhanced decay was found to be the most facile strategy to render ultra-pH response over the H-dimer and photoinduced electron transfer (PeT) mechanisms. Based on this insight, we selected several fluorophores with small Stoke shifts (<40 nm) and established a panel of multicolored nanoparticles with wide emission range (500–820 nm) and different pH transitions. Each nanoparticle maintained the sharp pH response (ON/OFF < 0.25 pH unit) with corresponding pH transition point at pH 5.2, 6.4, 6.9, and 7.2. Incubation of a mixture of multicolored nanoparticles with human H2009 lung cancer cells demonstrated sequential activation of the nanoparticles inside endocytic compartments directly correlating with their pH transitions. This multicolored, pH-tunable nanoplatform offers exciting opportunities for the study of many important cell physiological processes, such as pH regulation and endocytic trafficking of subcellular organelles

Convergent Synthesis of Enantioenriched <i>ortho</i>-Fused Tricyclic Diketones via Catalytic Asymmetric Intramolecular Vinylogous Aldol Condensation

Herein, we describe a catalytic asymmetric intramolecular vinylogous aldol reaction by taking advantage of dual organocatalysts, which enables convergent synthesis of ortho-fused tricyclic diketones in excellent enantioselectivities and diastereoselectivities. Noteworthy is that the reaction stereoselectively forges three consecutive stereogenic carbon centers including a quaternary one. Density functional theory calculations reveal that the enantioselectivity was facilitated by a transannular hydrogen bonding between the protonated quinuclidine moiety of the chiral aminocatalyst and the diketone fragment of the substrate

Efficient suppression of secretory clusterin levels by polymer-siRNA nanocomplexes enhances ionizing radiation lethality in human MCF-7 breast cancer cells in vitro-2

<p><b>Copyright information:</b></p><p>Taken from "Efficient suppression of secretory clusterin levels by polymer-siRNA nanocomplexes enhances ionizing radiation lethality in human MCF-7 breast cancer cells in vitro"</p><p></p><p>International Journal of Nanomedicine 2006;1(2):155-162.</p><p>Published online Jan 2006</p><p>PMCID:PMC2426783.</p><p>© 2006 Dove Medical Press Limited. All rights reserved</p

Efficient suppression of secretory clusterin levels by polymer-siRNA nanocomplexes enhances ionizing radiation lethality in human MCF-7 breast cancer cells in vitro-3

<p><b>Copyright information:</b></p><p>Taken from "Efficient suppression of secretory clusterin levels by polymer-siRNA nanocomplexes enhances ionizing radiation lethality in human MCF-7 breast cancer cells in vitro"</p><p></p><p>International Journal of Nanomedicine 2006;1(2):155-162.</p><p>Published online Jan 2006</p><p>PMCID:PMC2426783.</p><p>© 2006 Dove Medical Press Limited. All rights reserved</p