The use of data independent acquisition based proteomic analysis and machine learning to reveal potential biomarkers for autism spectrum disorder.

Autism spectrum disorder (ASD) is a complex neurological developmental disorder in children, and is associated with social isolation and restricted interests. The etiology of this disorder is still unknown. There is neither any confirmed laboratory test nor any effective therapeutic strategy to diagnose or cure it. We performed data independent acquisition (DIA) and multiple reaction monitoring (MRM) analysis of plasma from children with ASD and controls. The result showed that 45 differentially expressed proteins (DEPs) were identified between autistic subjects and controls. Among these, only one DEP was down-regulated in ASD; other DEPs were up-regulated in ASD children's plasma. These proteins are found associated with complement and coagulation cascades, vitamin digestion and absorption, cholesterol metabolism, platelet degranulation, selenium micronutrient network, extracellular matrix organization and inflammatory pathway, which have been reported to be related to ASD. After MRM verification, five key proteins in complement pathway (PLG, SERPINC1, and A2M) and inflammatory pathway (CD5L, ATRN, SERPINC1, and A2M) were confirmed to be significantly up-regulated in ASD group. Through the screening of machine learning model and MRM verification, we found that two proteins (biotinidase and carbonic anhydrase 1) can be used as early diagnostic markers of ASD (AUC = 0.8, p = 0.0001). SIGNIFICANCE: ASD is the fastest growing neurodevelopmental disorder in the world and has become a major public health problem worldwide. Its prevalence has been steadily increasing, with a global prevalence rate of 1%. Early diagnosis and intervention can achieve better prognosis. In this study, data independent acquisition (DIA) and multiple reaction monitoring (MRM) analysis was applied to analyze the plasma proteome of ASD patients (31 (±5) months old), and 378 proteins were quantified. 45 differentially expressed proteins (DEPs) were identified between the ASD group and the control group. They mainly were associated with platelet degranulation, ECM proteoglycar, complement and coagulation cascades, selenium micronutrient network, regulation of insulin-like growth factor (IGF) transport and uptake by insulin-like growth factor binding proteins (IGFBPs), cholesterol metabolism, vitamin metabolism, and inflammatory pathway. Through the integrated machine learning methods and the MRM verification of independent samples, it is considered that biotinidase and carbon anhydrase 1 have the potential to become biomarkers for the early diagnosis of ASD. These results complement proteomics database of the ASD patients, broaden our understanding of ASD, and provide a panel of biomarkers for the early diagnosis of ASD.

The protective effects of Esculentoside A through AMPK in the triple transgenic mouse model of Alzheimer's disease.

BACKGROUND: Neurofibrillary tangles comprising hyperphosphorylated tau are vital factors associated with the pathogenesis of Alzheimer's disease (AD). The elimination or reduction of hyperphosphorylated and abnormally aggregated tau is a valuable measure in AD therapy. Esculentoside A (EsA), isolated from Phytolacca esculenta, exhibits pharmacotherapeutic efficacy in mice with amyloid beta-induced AD. However, whether EsA affects tau pathology and its specific mechanism of action in AD mice remains unclear. PURPOSE: To investigate the roles and mechanisms of EsA in cognitive decline and tau pathology in a triple transgenic AD (3 × Tg-AD) mouse model. METHODS: EsA (5 and 10 mg/kg) was administered via intraperitoneal injection to 8-month-old AD mice for eight consecutive weeks. Y-maze and novel object recognition tasks were used to evaluate the cognitive abilities of mice. Potential signaling pathways and targets in EsA-treated AD mice were assessed using quantitative proteomic analysis. The NFT levels and hippocampal synapse numbers were investigated using Gallyas-Braak silver staining and transmission electron microscopy, respectively. Western blotting and immunofluorescence assays were used to measure the expression of tau-associated proteins. RESULTS: EsA administration attenuated memory and recognition deficits and synaptic damage in AD mice. Isobaric tags for relative and absolute quantitation proteomic analysis of the mouse hippocampus revealed that EsA modulated the expression of some critical proteins, including brain-specific angiogenesis inhibitor 3, galectin-1, and Ras-related protein 24, whose biological roles are relevant to synaptic function and autophagy. Further research revealed that EsA upregulated AKT/GSK3ß activity, in turn, inhibited tau hyperphosphorylation and promoted autophagy to clear abnormally phosphorylated tau. In hippocampus-derived primary neurons, inhibiting AMP-activated protein kinase (AMPK) activity through dorsomorphin could eliminate the effect of EsA, as revealed by increased tau hyperphosphorylation, downregulated activity AKT/GSK3ß, and blocked autophagy. CONCLUSIONS: To our knowledge, this study is the first to demonstrate that EsA attenuates cognitive decline by targeting the pathways of both tau hyperphosphorylation and autophagic clearance in an AMPK-dependent manner and it shows a high reference value in AD pharmacotherapy research.

Se-Methylselenocysteine (SMC) Improves Cognitive Deficits by Attenuating Synaptic and Metabolic Abnormalities in Alzheimer's Mice Model: A Proteomic Study.

Se-methylselenocysteine (SMC) is a major selenocompound in selenium (Se) enriched plants and has been found to ameliorate neuropathology and cognitive deficits in triple-transgenic mice model of Alzheimer's disease (3 × Tg-AD mice). To explore the underlying molecular mechanisms, the present study is designed to elucidate the protein changes in the cortex of SMC-treated 3 × Tg-AD mice. After SMC supplementation, proteomic analysis revealed that 181, 271, and 41 proteins were identified as differentially expressed proteins (DEPs) between 3 × Tg-AD mice vs wild type (AD/WT group), SMC-treated AD mice vs AD (AD + SMC/AD), and AD + SMC/WT group, respectively. Among these, 138 proteins in the diseased group were reversed by SMC treatment. The DEPs in AD/WT group and AD + SMC/AD group were mainly related to metabolism, synapses, and antioxidant proteins, while their levels were decreased in AD mice but up-regulated after treating with SMC. In addition, we found reduced ATP levels and destroyed synaptic structures in the AD mice brains, which were significantly ameliorated upon SMC treatment. Our study suggests that energy metabolism disorders, abnormal amino acid metabolism, synaptic dysfunction, and oxidative stress may be the key pathogenic phenomena of AD. SMC reversed the expression of proteins associated with them, which might be the main mechanism of its intervention in AD.

Exopolysaccharides from a Codonopsis pilosula endophyte activate macrophages and inhibit cancer cell proliferation and migration.

BACKGROUND: Exopolysaccharides with structural diversity have shown wide applications in biomaterial, food, and pharmaceutical industries. Herein, we isolated an endophytic strain, 14-DS-1, from the traditional medicinal plant Codonopsis pilosula to elucidate the characteristics and anti-cancer activities of purified exopolysaccharides. METHODS: HPLC and GC-MS were conducted to purify and characterize the exopolysaccharides isolated from 14-DS-1. Quantitative RT-PCR, cell migration assays, immunofluorescence staining, and flow cytometry analysis were conducted to investighate the biological activity of DSPS. RESULTS: We demonstrated that exopolysaccharides isolated from 14-DS-1 (DSPS), which were predominately composed of six monosaccharides, showed anti-cancer activities. Biological activity analysis revealed that exposure to DSPS induced macrophage activation and polarization by promoting the production of TNF-α and nitric oxide. Further analysis revealed that DSPS treatment promoted macrophage infiltration, whereas cancer cell migration was suppressed. In addition, DSPS exposure led to S-phase arrest and apoptosis in cancer cells. Immunofluorescence staining revealed that treatment with DSPS resulted in defects in spindle orientation and positioning. CONCLUSION: These findings thus suggest that DSPS may have promising potential in cancer therapy.

[Influence of photoselective vaporization of the prostate on the sexual function of the patient].

n recent years, photoselective vaporization of the prostate (PVP) has gained a wide clinical application in the treatment of benign prostatic hyperplasia (BPH) for its satisfactory effect, high safety, and low incidence of complications. With the improvement of living conditions, BPH patients are paying more attention to their sexual function, especially erectile function and ejaculatory problems instead of just focusing on the alleviation of lower urinary tract symptoms. Few studies of PVP, however, relate to its association with the sexual function of the patient and there is a certain controversy over the influence of PVP on it in the existing literature. Prevailing views hold that the uprated power in PVP does not affect erectile function or increase the risk of retrograde ejaculation (REj) and that PVP is even better than transurethral resection of the prostate (TURP) in avoiding the risk of REj.

Synthesis and Anti-Inflammatory Evaluation of Novel C66 Analogs for the Treatment of LPS-Induced Acute Lung Injury.

We previously reported a symmetric monocarbonyl analog of curcumin (MACs), C66, which demonstrated potential anti-inflammatory activity and low toxicity. In continuation of our ongoing research, we designed and synthesized 34 asymmetric MACs based on C66 as a lead molecule. A majority of the C66 analogs effectively inhibited LPS induction of TNF-α and IL-6 expression. Additionally, a preliminary SAR was conducted. Furthermore, active compounds 4a11 and 4a16 were found to effectively reduce the W/D ratio in the lungs and the protein concentration in the bronchoalveolar lavage fluid (BALF). Meanwhile, a histopathological examination indicated that these two analogs significantly attenuate tissue injury in the lungs with LPS-induced ALI rats. 4a11 and 4a16 also inhibited mRNA expression of several inflammatory cytokines, including TNF-α, IL-6, IL-1ß, COX-2, ICAM-1 and VCAM-1, in the Beas-2B cells after LPS challenge. Altogether, the data exhibit a series of new C66 analogs as promising anti-inflammatory agents for the treatment of LPS-induced ALI.

Electrochemical oxidation and detection of paeonol on modified electrode with acetylene black nanoparticles.

With an aim to construct a sensing platform for the electrochemical detection of paeonol, we modified the glassy carbon electrode with acetylene black nanoparticle (AB). A sensitive oxidation peak of paeonol was observed with remarkably increased peak current on the modified electrode because the electrode has a big surface area due to three dimensional structure of AB nanoparticles. The optimization of detection conditions was performed, including pH value of the buffer, the amount of AB nanoparticles on the electrode surface, the accumulation potential and time of paeonol. Under the optimized conditions, the oxidation peak current of paeonol increased linearly with its concentration over the range from 5×10(-7) to 1×10(-4) M. The detection limit was calculated to be 1×10(-7) M. The modified electrode was successfully applied to detect the content of paeonol in cortex moutan, a common traditional Chinese medicine. The method is new, sensitive, rapid and convenient for the detection of paeonol.

Electrochemical detection of honokiol and magnolol in traditional Chinese medicines using acetylene black nanoparticle-modified electrode.

INTRODUCTION: Honokiol and magnolol are the active components of Magnolia officinalis, which is a widely used traditional Chinese medicine. Their simultaneous analysis is, therefore, important for the quality control of the product. OBJECTIVE: To establish a simple, sensitive and rapid electrochemical method for the simultaneous detection of honokiol and magnolol based on the remarkable enhancement effect of acetylene black nanoparticle (AB). METHODOLOGY: The AB-modified electrode was prepared via solvent evaporation. The electrochemical response of honokiol and magnolol was investigated using cyclic voltammetry. The simultaneous detection was performed with differential pulse voltammetry. The method was validated in terms of linearity, sensitivity, precision and accuracy. RESULTS: The linear range for honokiol is 0.5-300 µg/L, and the limit of detection (LOD) is 0.25 µg/L (9.4 × 10(-10) mol/L). For magnolol, the linear range is 10-250 µg/L, and the LOD is 5 µg/L (1.88 × 10(-8) mol/L). CONCLUSION: The new method was successfully used to determine honokiol and magnolol in a traditional Chinese medicine called Ageratum liquid.