Identification and characterization of Tu88, an antigenic gene from Theileria uilenbergi.

Theileria uilenbergi is a pathogen that causes ovine theileriosis. Prevention and control of theileriosis relies on its diagnosis at early stages of occurrence and requires understanding of proteins with antigenic properties from the pathogen. Despite its prevalence in China, only a few molecules with antigenic properties have been characterized from T. uilenbergi. In this study, we identified a cDNA named Tu88 by immunoscreening a T. uilenbergi merozoite cDNA library with T. uilenbergi-positive sera from infected sheep. Recombinant Tu88 (rTu88) expressed in bacteria reacted strongly with the positive sera of T. uilenbergi in western blot analysis indicating its potential as an antigen. Southern blot analysis showed that it is a single copy gene. Protein localization by immunostaining blood smears from an infected sheep demonstrated the presence of native Tu88 in merozoites. These findings suggest that Tu88 is a potential candidate antigen for the development of a sero-diagnostic tool.

In Silico Prediction of Quercetin Analogs for Targeting Death-Associated Protein Kinase 1 (DAPK1) Against Alzheimer's Disease.

Alzheimer's Disease (AD) is a progressive neurodegenerative disorder that greatly affects the health and life quality of the elderly population. Existing drugs mainly alleviate symptoms but fail to halt disease progression, underscoring the urgent need for the development of novel drugs. Based on the neuroprotective effects of flavonoid quercetin in AD, this study was designed to identify potential AD-related targets for quercetin and perform in silico prediction of promising analogs for the treatment of AD. Database mining suggested death-associated protein kinase 1 (DAPK1) as the most promising AD-related target for quercetin among seven protein candidates. To achieve better biological effects for the treatment of AD, we devised a series of quercetin analogs as ligands for DAPK1, and molecular docking analyses, absorption, distribution, metabolism, and excretion (ADME) predictions, as well as molecular dynamics (MD) simulations, were performed. The energy for drug-protein interaction was predicted and ranked. As a result, quercetin-A1a and quercetin-A1a1 out of 19 quercetin analogs exhibited the lowest interaction energy for binding to DAPK1 than quercetin, and they had similar dynamics performance with quercetin. In addition, quercetin-A1a and quercetin-A1a1 were predicted to have better water solubility. Thus, quercetin-A1a and quercetin-A1a1 could be promising agents for the treatment of AD. Our findings paved the way for further experimental studies and the development of novel drugs.

Clinical epidemiology survey of the traditional Chinese medicine etiology and syndrome differentiation of coronary artery disease: study protocol of a multicenter trial.

BACKGROUND: Coronary artery disease (CAD), a common disease with high incidence and mortality rate, has seriously threatened the health and life of the public. Traditional Chinese medicine (TCM) has an important role in the prevention and treatment of this disease. Through clinical epidemiological survey, a deeper understanding of TCM etiology and syndrome characteristics in CAD would further improve clinical efficacy in the treatment of this disease. METHODS/DESIGN: The preliminary clinical questionnaire for TCM etiology and syndrome differentiation in CAD was designed after literature reviews and analysis. Through a series of clinical pre-surveys, expert consultation and demonstration, the formal TCM clinical epidemiology questionnaire on the etiology and syndrome differentiation in CAD was finalized, after which, the study protocol, inclusive and exclusive criteria and related quality control measures were prepared. The multiregional clinical epidemiological survey with more than 5000 participants with CAD will be carried out in 41 TCM hospitals of China for investigating the TCM etiology and syndrome differentiation of CAD. DISCUSSION: Multiregion large sample size clinical epidemiology survey on TCM etiology and syndrome differentiation in CAD will provide further evidence in preventing CAD and improving the standardization process of syndrome research. TRIAL REGISTRATION: This study protocol was registered at the Chinese Clinical Trial Registry on November 27, 2011 in both Chinese and English editions and the registration number is ChiCTR-ECS-11001728.

Protective Role of Shiitake Mushroom-Derived Exosome-Like Nanoparticles in D-Galactosamine and Lipopolysaccharide-Induced Acute Liver Injury in Mice.

Fulminant hepatic failure (FHF) is a rare, life-threatening liver disease with a poor prognosis. Administration of D-galactosamine (GalN) and lipopolysaccharide (LPS) triggers acute liver injury in mice, simulating many clinical features of FHF in humans; therefore, this disease model is often used to investigate potential therapeutic interventions to treat FHF. Recently, suppression of the nucleotide-binding domain and leucine-rich repeat related (NLR) family, pyrin domain containing 3 (NLRP3) inflammasome, was shown to alleviate the severity of GalN/LPS-induced liver damage in mice. Therefore, the goal of this study was to find dietary exosome-like nanoparticles (ELNs) with therapeutic potential in curbing FHF by suppressing the NLRP3 inflammasome. Seven commonly consumed mushrooms were used to extract ELNs. These mushrooms were found to contain ELNs composed of RNAs, proteins, and lipids. Among these mushroom-derived ELNs, only shiitake mushroom-derived ELNs (S-ELNs) substantially inhibited NLRP3 inflammasome activation by preventing inflammasome formation in primary macrophages. S-ELNs also suppressed the secretion of interleukin (IL)-6, as well as both protein and mRNA levels of the Il1b gene. Remarkably, pre-treatment with S-ELNs protected mice from GalN/LPS-induced acute liver injury. Therefore, S-ELNs, identified as potent new inhibitors of the NLRP3 inflammasome, represent a promising class of agents with the potential to combat FHF.

Development of an immunochromatographic strip for the serodiagnosis of Theileria infection in sheep.

BACKGROUND: Theileria uilenbergi and T. luwenshuni are tick-borne protozoan parasites, transmitted by Haemaphysalis qinghaiensis and H. longicornis, respectively. They are the main causative agents of theileriosis in small ruminants in China. The disease has resulted in severe economic losses and hindered the development of sheep and goat husbandry industry in the endemic regions. METHODS: In this study, a colloidal gold-based immunochromatographic strip (ICS) was developed for the detection of T. uilenbergi and/or T. luwenshuni infections. A recombinant T. uilenbergi immunodominant protein (rTuIP) was used as antigen for the ICS. The nitrocellulose membrane was incubated with rTuIP on the test (T) line and anti-rTuIP antiserum on the control (C) line, respectively. The rTuIP conjugated to colloidal gold particles was used as the detection system for visualization of the lines. Then the sample pad, conjugate pad, nitrocellulose membrane and absorbent pad were assembled onto a backing plate in the appropriate order. RESULTS: The ICS was able to detect antibodies in the sera of animals experimentally infected with T. uilenbergi from 14 to 85 days. It also reacted with the serum from T. luwenshuni infected sheep. However, there was no cross-reactivity with sera from animals infected with Babesia motasi and Anaplasma ovis. Comparison of the ICS with the rTuIP antigen based indirect enzyme-linked immunosorbent assays (ELISA) using test field samples showed good correlations with 93.1 % (81/87) sensitivity and 100 % (40/40) specificity, respectively, with an almost perfect agreement (Kappa = 0.895, P < 0.01). CONCLUSION: An immunochromatographic strip test based on a recombinant T. uilenbergi immunodominant protein (rTuIP) was developed. This is a rapid test (approximately 15 min to completion) for the detection of T. uilenbergi and/or T. luwenshuni infection that is easy to perform and; delivers results that are visible to the naked eye.