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Objective: To identify potential serum protein candidates involved in linking the traditional Chinese medicine (TCM)-defined qi deficiency constitution (QDC) to Pi-qi-deficiency syndrome (PQDS) of chronic superficial gastritis (CSG). Methods: Using participants with the TCM-defined balanced constitution as a control population, label-free quantitative proteomics was adopted to identify differentially expressed proteins (DEPs) in serum samples from two case populations: case population 1 (participants with QDC) and case population 2 (patients with PQDS of CSG). The DEPs discovered in both case populations were analyzed to identify common DEPs as potential candidates for proteins involved in the link between QDC and PQDS. Based on Kyoto Encyclopedia of Genes and Genomes pathway (KEGG) and Gene Ontology (GO) enrichment analysis and analysis of protein–protein interaction networks, we evaluated the possible functions of these potential serum candidates. Results: We discovered 24 and 28 proteins that were differentially expressed in case populations 1 and 2, respectively, compared with the control population. Hierarchical clustering analysis showed that the expression profile of DEPs of individuals from the same population clustered well, while those from different populations were segregated. Furthermore, GO analysis revealed the 10 DEPs that were common to both case populations to be mainly associated with negative regulation of cellular metabolic and immune system processes while KEGG analysis indicated these proteins to be associated with complement and coagulation cascades and peroxisome proliferator-activated receptor signaling. Notably, serum levels of C4b-binding protein beta chain, glycosylphosphatidylinositol-specific phospholipase D1 and MS-F1 light chain variable region proteins were notably higher in the two case populations compared with the control, particularly in the case of CSG with PQDS. Conclusion: The results presented here provide new insights into the molecular mechanisms underlying development of PQDS of CSG from QDC, and suggest candidate serum biomarkers for future application in integrative medicine. Keywords: Qi deficiency constitution, Pi-qi-deficiency syndrome, Chronic superficial gastritis, Serum biomarker

The applications of viral protein cages have expanded rapidly into the fields of bionanotechnology and materials science. However, the low-cost production of viral capsid proteins (CPs) on a large scale is always a challenge. Herein, we develop a highly efficient expression system by constructing recombinant Pichia pastoris cells as a “factory” for the secretion of soluble cowpea chlorotic mottle virus (CCMV) CPs. Under optimal induction conditions (0.9 mg/mL of methanol concentration at 30 °C for 96 h), a high yield of approximately 95 mg/L of CCMV CPs was harvested from the fermentation supernatant with CPs purity >90%, which has significantly simplified the rest of the purification process. The resultant CPs are employed to encapsulate Ruthenium (Ru) nanoparticles (NPs) via in-vitro self-assembly to prepare hybrid nanocatalyst, i.e. Ru@virus-like particles (VLPs). The catalytic activity over Ru@VLPs was evaluated by reducing 4-nitrophenol (4-NP) to 4-aminophenol (4-AP). The results indicate that, with the protection of protein cages, Ru NPs were highly stabilized during the catalytic reaction. This results in enhanced catalytic activity (reaction rate constant k = 0.14 min−1) in comparison with unsupported citrate-stabilized Ru NPs (Ru-CA) (k = 0.08 min−1). Additionally, comparatively lower activation energy over Ru@VLPs (approximately 32 kJ/mol) than that over Ru-CA (approximately 39 kJ/mol) could be attributed to the synergistic effect between Ru NPs and some functional groups such as amino groups (–NH2) on CPs that weakened the activation barrier of 4-NP reduction. Therefore, enhanced activity and decreased activation energy over Ru@VLPs demonstrated the superiority of Ru@VLPs to unsupported Ru-CA.

The thermal energy storage performance and phase change stability of mix phase change materials consisted of stearic acid and Na2PO4·12H2O are studied by means of Fourier transformation infrared spectrum analyses (FTIR) and Differential Scanning Calorimetry (DSC). The specific heat capacity of mix materials is also determined experimentally. The results show that all absorption peaks of functional groups and chemical bonds of stearic acid still exist, which suggests that these crystal materials composed of stearic acid and Na2PO4·12H2O have good phase change stability and chemical compatibility. Thus, disadvantages caused by the subcooling of Na2PO4·H2O and the low specific heat capacity of stearic acid can be solved, it is speculated that complement each other on the basis of organic/inorganic characteristics in itself is carried out for these two component materials of the mixture.

Cytotoxic T cells (CTLs) are an important component of adaptive immunity. The study of antigen-specific CTLs in vivo is desirable yet difficult. Identification of the class I-restricted peptide used by CTLs for target recognition is often required for detailed studies, but is generally not known for most antigens. Toxoplasma gondii is a medically important, obligate intracellular parasite and is often used as a model for studies of parasite immunology. No class I-restricted peptides for CTLs are known. We show here a new and convenient method to detect T. gondii-specific CTLs in vivo. We engineered T. gondii tachyzoites to express the model antigen ovalbumin, for which many useful reagents and transgenic mice are available. Using ovalbumin-transgenic T. gondii tachyzoites, antigen-specific CTLs were detected in vivo, and at much earlier time points post-infection than previously reported. This new method has several additional advantages over current methods to detect T. gondii-specific CTLs.

Background Cancer prevention is a cost-effective alternative to treatment. Oral rapamycin (eRapa) prevents distinct cancers and extends life in mice [1] , making it a candidate broad-spectrum cancer prevention agent. Rapamycin, an inhibitor of mammalian target of rapamycin (mTOR), has significant immune effects that are unstudied in cancer. We hypothesize that eRapa prevents cancer through improved cancer immune surveillance, which we tested in a carcinogen-induced, inflammation-driven skin cancer model. Methods eRapa was used at 14 ppm microencapsulated rapamycin. Skin cancer. Wild-type C57BL/6 (WT) or syngeneic β δ knockout (KO) and interferon (IFN)- γ KO mice (lacking all T cells or IFN- γ , respectively) got eRapa or control for 1 month, followed once with the carcinogen 7,12-dimethylbenz[a]anthracene (DMBA), then twice weekly applications with the inflammatory agent 12-O-tetradecanoylphorbol (TPA) for 24 weeks. Flow cytometry assessed cells and IFN- γ . Statistics. Tumors (unpaired t test and 2-way ANOVA) and malignant degeneration (Fischer exact test). Results eRapa reduced benign ( p = .004) and malignant ( p = .03) tumors in WT mice. T cells and IFN- γ mediate cancer immune surveillance so their effects on eRapa cancer prevention were studied. eRapa reduced skin tumors in β δ KO mice lacking all T cells ( p = .04), but not in IFN- γ KO mice ( p = .13), consistent with loss of beneficial eRapa-induced, non-T cell IFN- γ . In support, WT or IFN- γ KO T cell transfer into IFN- γ KO mice did not alter eRapa cancer prevention in this model. In WT mice on DMBA/TPA, eRapa increased IFN- γ producing dermal natural killer (NK) cells ( p γ KO mice, suggesting that eRapa-mediated IFN- γ facilitated local NK cell accumulation. Conclusions eRapa prevents carcinogen and inflammation-induced skin cancer, which involves a non-T cell that could be an NK cell, and is IFN- γ dependent. We also show that eRapa prevents cancer and extends life in distinct mouse cancer models, which paves the way for use of mTOR inhibitors as broad spectrum human cancer prevention agents.

Chloroquine has been widely used as a reagent to enhance transfection in many non-viral gene delivery systems. The mechanism of its action is not well understood and has not been extensively investigated. We synthesized a number of CQ analogues with variations either on the alkyl amino side chain or on the aromatic ring, and investigated their structure-function correlations as transgene expression enhancers. We found that the aliphatic amino moiety is essential to CQ's role as a gene expression enhancer, as its removal eliminates any enhancement. Further, the enhancement is dramatically affected by changes to the aromatic ring, and is positively correlated to interactions between DNA and CQ analogues. Quinacrine, a CQ analogue with a tricyclic aromatic acridinyl structure that can strongly intercalate DNA, can enhance transfection similarly to CQ at a concentration ten times lower. In contrast, N4-(7-Chloro-4-pyridinyl)-N1,N1-diethyl-1,4-pentanediamine, a CQ analogue that has a weak intercalating pyridinyl ring, has no effect on gene expression. Subtle change of the 7-substituent of the CQ aromatic structure can also greatly affect its ability to enhance transgene expression. Transfection in the presence of N4-(7-trifluoromethyl-4-quinolinyl)-N1,N1-diethyl-1,4-pentanediamine (TFCQ), a 7- trifluoromethyl CQ analogue, shows expression at least one order of magnitude higher than transfection in the presence of CQ at the same concentration, while transfection in the presence of N4-(4-quinolinyl)-N1,N1-diethyl-1,4-pentanediamine, a 7-hydro CQ analogue, shows no enhancement in expression. CQ and TFCQ demonstrate similar activity in buffering the pH experienced by polyplexes within the endocytic pathway, but TFCQ is more potent in enhancing gene transfer. TFCQ has higher binding affinity to DNA than CQ as well as a greater tendency to competitively displace the polycation from polyplexes at an achievable intracellular concentration. These findings suggest that in addition to pH-buffering activity, CQ's competitive displacement of polycations and its interaction with nucleic acids are important to its enhancement of gene expression in non-viral nucleic acid delivery systems. Through a number of comparative studies on CQ and its analogues, we elucidated for the first time the important structural features of CQ that confer its capability to accumulate inside cells, buffer intracellular pH, and competitively displace polycations and thus unpackage DNA.

A widely applicable cancer prevention strategy would be a cost-effective approach to lessen the enormous burden of cancer. Enterically-released microencapsulated rapamycin (eRapa) extends lifespan in mice and delays or prevents cancer development, suggesting cancer prevention as a longevity mechanism. Rapamycin acts through inhibition of mechanistic target of rapamycin (mTOR), which has significant immune effects that are surprisingly little studied in cancer treatment or prevention. In naive mice, chronic eRapa significantly reduced Th1 (CXCR3 + ) cell numbers ( p = .03) and T cell interferon (IFN)- γ ( p = .04), increased Th17 (CCR6 + ) cell numbers ( p = .04) and T cell IL-17 ( p = .03), increased T cell IL-22 ( p = .02), and reduced immune suppressing regulatory T cells ( p βδ −/− and IFN- γ −/− mice (lacking all T cells or IFN- γ , respectively) were given eRapa or control, and skin tumors were induced with the carcinogen 7,12-dimethylbenz[a]anthracene (DMBA) followed by the inflammatory promoting agent 12-O-tetradecanoylphorbol-13-acetate (TPA). eRapa reduced papilloma size in WT BL6 mice ( p = .02) versus controls, the first demonstration that oral rapamycin prevents inflammation-mediated neoplasia. eRapa also significantly reduced papillomas in βδ −/− mice ( p = .04) and increased papillomas in IFN- γ −/− mice ( p = .05). These data are consistent with the concept that eRapa protects from carcinogen-induced dermal carcinogenesis through a non-T cell, IFN- γ -dependent mechanism, which we suspect includes natural killer cells. We have also shown that eRapa prevents cancer in a variety of mouse cancer models, in which we are further studying immune mechanisms. Immune contributions from mTOR inhibitors in cancer prevention (and treatment) require more attention.