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A new immunomodulatory protein, designated ACA, was purified from the mycelium extract of Antrodia camphorata , a well-known folk medicine bitter mushroom in Taiwan, and N-terminally sequenced. By taking advantage of its N-terminal amino acid sequence, the full-length ACA gene was cloned using rapid amplification of cDNA ends (RACE) approach. This gene encodes a 136 amino acid protein that is homologous to the phytotoxic proteins from fungi. On the basis of the data of N-terminal sequencing and N-glycosidase F treatment, the native ACA was confirmed to be a glycoprotein. The similarity in activation of TLR4-deficient macrophages by both the native ACA and recombinant ACA (rACA) suggested that the glycosyl group(s) of the native ACA was insignificant in macrophage activation. Moreover, the failure of rACA to induce TLR2-deficient macrophages and to activate the RAW 264.7 macrophages transfected with the dominate-negative MyD88 (dnMyD88) indicated that the ACA-mediated macrophage activation was TLR2/MyD88 dependent. Microarray assay of the ACA-activated NFκB-related gene expression showed that rACA demonstrated a LPS-mimetic proinflammatory response toward RAW 264.7 macrophages. Furthermore, rACA enhanced phagocytosis activity and CD86 (B7-2) expression as well as induced TNF-α and IL-1β production within murine peritoneal macrophages. A time-dependent induction of mRNA expression of cytokines TNF-α, IL-1β, IL-6, and IL-12 as well as chemokines CCL3, CCL4, CCL5, and CCL10, but not IL-10, CCL17, CCL22, and CCL24, was observed after the ACA treatment of the macrophages. These results proposed that ACA exhibited M1 polarization and differentiation in macrophages. Thus, ACA is an important immunomodulatory protein of A. camphorata.

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The development of noninvasive methods for real-time in vivo analysis is of great significant, which provides powerful tools for medical research and clinical diagnosis. In the present work, we described a new strategy based on cataluminescence (CTL) for real-time in vivo clinical breath analysis. To illustrate such strategy, a homemade real-time CTL monitoring system characterized by coupling an online sampling device with a CTL sensor for sevoflurane (SVF) was designed, and a real-time in vivo method for the monitoring of SVF in exhaled breath was proposed. The accuracy of the method was evaluated by analyzing the real exhaled breath samples, and the results were compared with those obtained by GC/MS. The measured data obtained by the two methods were in good agreement. Subsequently, the method was applied to real-time monitoring of SVF in exhaled breath from rat models of the control group to investigate elimination pharmacokinetics. In order to further probe the potential of the method for clinical application, the elimination pharmacokinetics of SVF from rat models of control group, liver fibrosis group alcohol liver group, and nonalcoholic fatty liver group were monitored by the method. The raw data of pharmacokinetics of different groups were normalized and subsequently subjected to linear discriminant analysis (LDA). These data were transformed to canonical scores which were visualized as well-clustered with the classification accuracy of 100%, and the overall accuracy of leave-one-out cross-validation procedure is 88%, thereby indicating the utility of the potential of the method for liver disease diagnosis. Our strategy undoubtedly opens up a new door for real-time clinical analysis in a pain-free and noninvasive way and also guides a promising development direction for CTL.

Rosin derived from conifer trees is used as the basis for a novel environmentally-friendly adsorbent prepared from a sustainable resource. After treatment with ethylenediamine, ethylenediamine rosin-based resin (EDAR) is produced, which possesses cation exchange capacity that is comparable to that of the best commercial synthetic resins. This is demonstrated by its application to the removal of Pb, Cd, and Cu from water, in single and multicomponent systems. Maximum uptake was obtained at pH 5 and in the order Pb(II) > Cd(II) > Cu(II). The maximum adsorption of Pb was ~1.8 mmol/g, but the adsorption process resembled the Freundlich isotherm, whereas the adsorption of Cd(II) and Cu(II) followed the Langmuir isotherm. In the multicomponent systems, there was direct competition between Pb and Cd for sorption sites, whereas the results with Cu indicated it had a preference for different types of sites compared to Pb and Cd. The EDAR resin could be efficiently regenerated and used repeatedly with only a small decrease in performance. Characterization of EDAR, and investigations of its adsorption mechanisms using physical, spectroscopic, and theoretical techniques, including fourier transform infrared spectroscopy (FTIR), 13C nuclear magnetic resonance (13C NMR), scanning electron microscope (SEM), Brunauer Emmett Teller (BET) method, elemental analysis, thermogravimetric analysis (TGA), and molecular dynamics calculations, showed that amino groups have a critical role in determining the cation adsorption properties. We conclude that this new adsorbent derived from an abundant natural material has the potential to make valuable contributions to the routine removal of heavy metal ions (HMs) from drinking water and wastewater.