Transplantation of Wnt5a-modified Bone Marrow Mesenchymal Stem Cells Promotes Recovery After Spinal Cord Injury via the PI3K/AKT Pathway.

Spinal cord injury (SCI) is a severe neurological condition that can lead to paralysis or even death. This study explored the potential benefits of bone marrow mesenchymal stem cell (BMSC) transplantation for repairing SCI. BMSCs also differentiate into astrocytes within damaged spinal cord tissues hindering the cell transplantation efficacy, therefore it is crucial to enhance their neuronal differentiation rate to facilitate spinal cord repair. Wnt5a, an upstream protein in the non-classical Wnt signaling pathway, has been implicated in stem cell migration, differentiation, and neurite formation but its role in the neuronal differentiation of BMSCs remains unclear. Thus, this study investigated the role and underlying mechanisms of Wnt5a in promoting neuronal differentiation of BMSCs both in vivo and in vitro. Wnt5a enhanced neuronal differentiation of BMSCs in vitro while reducing astrocyte differentiation. Additionally, high-throughput RNA sequencing revealed a correlation between Wnt5a and phosphoinositide 3-kinase (PI3K)/protein kinase B(AKT) signaling, which was confirmed by the use of the PI3K inhibitor LY294002 to reverse the effects of Wnt5a on BMSC neuronal differentiation. Furthermore, transplantation of Wnt5a-modified BMSCs into SCI rats effectively improved the histomorphology (Hematoxylin and eosin [H&E], Nissl and Luxol Fast Blue [LFB] staining), motor function scores (Footprint test and Basso-Beattie-Bresnahan [BBB]scores)and promoted neuron production, axonal formation, and remodeling of myelin sheaths (microtubule associated protein-2 [MAP-2], growth-associated protein 43 [GAP43], myelin basic protein [MBP]), while reducing astrocyte production (glial fibrillary acidic protein [GFAP]). Therefore, targeting the Wnt5a/PI3K/AKT pathway could enhance BMSC transplantation for SCI treatment.

Simultaneous determination of five constituents of areca nut extract in rat plasma using UPLC-MS/MS and its application in a pharmacokinetic study.

Areca nuts have been used as a traditional Chinese medicine (TCM) for thousands of years. Recent studies have shown that it exhibits good pharmacological activity and toxicity. In this study, the pharmacokinetics of five major components of areca nut extract in rats were investigated using a highly sensitive ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-MS/MS) method. Arecoline, arecaidine, guvacoline, guvacine, and catechin were separated and quantified accurately using gradient elution with mobile phases of (A) water containing 0.1 % formic acid-10 mM ammonium formate, and (B) methanol. The constituents were detected under a timing switch between the positive and negative ion modes using multiple reaction monitoring (MRM). Each calibration curve had a high R2 value of >0.99. The method accuracies ranged -7.09-11.05 % and precision values were less than 14.36 %. The recovery, matrix effect, selectivity, stability, and carry-over of the method were in accordance with the relevant requirements. It was successfully applied for the investigation of the pharmacokinetics of these five constituents after oral administration of areca nut extract. Pharmacokinetic results indirectly indicated a metabolic relationship between the four areca nut alkaloids in rats. For further clarification of its pharmacodynamic basis, this study provided a theoretical reference.

Tauroursodeoxycholic Acid Inhibited Apoptosis and Oxidative Stress in H2O2-Induced BMSC Death via Modulating the Nrf-2 Signaling Pathway: the Therapeutic Implications in a Rat Model of Spinal Cord Injury.

Spinal cord injury (SCI) is a prevalent and significant injury to the central nervous system, resulting in severe consequences. This injury is characterized by motor, sensory, and excretory dysfunctions below the affected spinal segment. Transplantation of bone marrow mesenchymal stem cells (BMSCs) has emerged as a potential treatment for SCI. However, the low survival as well as the differentiation rates of BMSCs within the spinal cord microenvironment significantly limit their therapeutic efficiency. Tauroursodeoxycholic acid (TUDCA), an active ingredient found in bear bile, has demonstrated its neuroprotective, antioxidant, and antiapoptotic effects on SCI. Thus, the present study was aimed to study the possible benefits of combining TUDCA with BMSC transplantation using an animal model of SCI. The results showed that TUDCA significantly enhanced BMSC viability and reduced apoptosis (assessed by Annexin V-FITC, TUNEL, Bax, Bcl-2, and Caspase-3) as well as oxidative stress (assessed by ROS, GSH, SOD, and MDA) both in vitro and in vivo. Additionally, TUDCA accelerated tissue regeneration (assessed by HE, Nissl, MAP2, MBP, TUJ1, and GFAP) and improved functional recovery (assessed by BBB score) following BMSC transplantation in SCI. These effects were mediated via the Nrf-2 signaling pathway, as evidenced by the upregulation of Nrf-2, NQO-1, and HO-1 expression levels. Overall, these results indicate that TUDCA could serve as a valuable adjunct to BMSC transplantation therapy for SCI, potentially enhancing its therapeutic efficacy.

Two-Omics Probe on the Potential of Pseudomonas sp. GDMCC 1.1703 Under Phenol Stress.

Pseudomonas sp. harbors genetic diversity and readily adapts to environmental challenges, conferring upon it the ability to remediate. It is important to genetically determine the effects of bacterial application. The two-omics integration approach may shed more light on Pseudomonas isolates, filling the knowledge gap between genetic potential and dynamic function. In the present study, a strain from the Xi River was isolated using benzene-selective enrichment medium and phylogenetically identified as Pseudomonas sp. GDMCC 1.1703 by 16S rRNA gene sequencing. Its phenol degradability was optimally assessed at a rate of 45.7% (by statistics P < 0.05) in 12 h with a 200 mg/L concentration. Genomics and transcriptomics analyses were successively used to identify the genes and pathways responsible for phenol degradation. At least 42 genes were genomically identified to be involved in xenobiotic biodegradation. The degradative genes clustered into operons were hypothesized to have evolved through horizontal gene transfer. On the basis of genomic authentication, transcriptome analysis dynamically revealed that phenol degradation and responsive mechanisms were both upregulated as defense between the Ctrl (control) and PS (phenol-stressed) groups. Quantitative reverse transcription-PCR not only validated the key genes identified via RNA sequencing but also consistently confirmed the realistic intracellular expression. The approach of omics integration, which is effective in exploring the potential of isolates, will hopefully become an established method for determining the remediation potential of a candidate for development.

In vitro bloodbrain barrier permeability study of four main active ingredients from Alpiniae oxyphyllae fructus.

The fruits of Alpinia oxyphylla Miq., a broadly utilized traditional Chinese medicine, have a number of effects on the central nervous system (CNS). The main active constituents of Alpiniae oxyphyllae fructus (AOF) were nootkatone, tectochrysin, chrysin and protocatechuic acid. An immortalized human brain microvascular endothelial cell (hCMEC/D3) and astrocyte (HA1800) coculture model was used to investigate the permeability of the blood-brain barrier (BBB). The validation of ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) methods for the four compounds was conducted following industry guidelines. Calibration curves were generated with mean coefficients (R2) better than 0.99. The inter-day and intra-day precisions were less than 8.53% and 7.12%, respectively. The accuracies were lower than ± 11.57%, and recoveries were greater than 86.07%. The samples of the transport experiment were examined, and the apparent permeability coefficients (Papp) were calculated. The efflux ratios of the four compounds are all less than 2. The Papp values of protocatechuic acid, chrysin, nootkatone, tectochrysin were at the level of 10-5, 10-6, 10-6, and 10-7 cm/s, respectively. All four compounds crossed the BBB by passive diffusion, with protocatechuic acid having high permeability, and tectochrysin having poor permeability. This research indicated the permeability of protocatechuic acid, chrysin, nootkatone and tectochrysin through the BBB and offered a foundation for related research on AOF in the treatment of CNS illnesses.

Associations of Leisure Activities and Sleep Duration with Cognitive Function: A Nationwide Prospective Cohort Study of Chinese Old Adults.

BACKGROUND: Leisure activities and sleep duration are correlated and have been linked to cognitive function, but most studies have examined only one of these factors. OBJECTIVE: To investigate the independent and joint associations of leisure activities and sleep duration with cognitive function among older adults. METHODS: We included 7,796 participants aged≥65 years from the Chinese Longitudinal Healthy Longevity Survey during 2008-2018 (waves 5-8). Self-reported leisure activities and sleep duration were assessed at baseline, and cognitive function was measured repeatedly using the Chinese version of the Mini-Mental State Examination (MMSE) at baseline and during follow-up. We used linear mixed models to estimate regression coefficients (ß) and 95% confidence intervals (CI). RESULTS: The median follow-up duration was 5.77 years. After adjusting for each other and potential confounders, both lower leisure activity score (each 1-point decrease ß= -0.33, 95% CI: -0.36 to -0.30) and longer sleep duration (each 1-hour increase ß= -0.17, 95% CI: -0.22 to -0.11) were independently associated with lower MMSE score. Furthermore, we observed an additive interaction between leisure activities and sleep duration (pinteraction < 0.001). A combination of low leisure activity score and long sleep duration was strongly associated with decreased MMSE score (ß= -2.51, 95% CI: -2.85 to -2.16) compared with the group with combined high leisure activity score and normal sleep duration. CONCLUSION: Both leisure activities and sleep duration were independently associated with cognitive function. Moreover, the combination of leisure inactivity and prolonged sleep duration predicted worse cognitive function (a preclinical hallmark of Alzheimer's disease) in an additive manner.

Integrated untargeted fecal metabolomics and gut microbiota strategy for screening potential biomarkers associated with schizophrenia.

Schizophrenia (SZ) is a serious neurodevelopmental disorder. As the etiology of SZ is complex and the pathogenesis is not thoroughly understood, the diagnosis of different subtypes still depends on the subjective judgment of doctors. Therefore, there is an urgent need to develop early objective laboratory diagnostic biomarkers to screen different subtypes of patients as early as possible, and to implement targeted prevention and precision medicine to reduce the risk of SZ and improve patients' quality of life. In this study, untargeted metabolomics and 16S rDNA sequencing were used to analyze the differences in metabolites and gut microflora among 28 patients with two types of schizophrenia and 11 healthy subjects. The results showed that the metabolome and sequencing data could effectively discriminate among paranoid schizophrenia patients, undifferentiated schizophrenia patients and healthy controls. We obtained 65 metabolites and 76 microorganisms with significant changes, and fecal metabolite composition was significantly correlated with the differential genera (|r|>0.5), indicating that there was a regulatory relationship between the gut microbiota and the host metabolites. The gut microbiome, as an objective and measurable index, showed good diagnostic value for distinguishing schizophrenia patients from healthy people, especially with a combination of several differential microorganisms, which had the best diagnostic effect (AUC>0.9). Our results are conducive to understanding the complicated metabolic changes in SZ patients and providing valuable information for the clinical diagnosis of SZ.

Correlation of phase angle with sarcopenia and its diagnostic value in elderly men with cancer.

OBJECTIVES: The aims of this study was to investigate the association of phase angle (PA) with sarcopenia and its components and to evaluate the effectiveness of PA in sarcopenia diagnosis in older men (>65 y of age) with cancer. METHODS: The study included older men with non-small cell lung cancer and digestive tract cancer who were hospitalized in the past 3 y. General characteristics such as age, body mass index, and tumor stage were gathered. Mid-upper arm muscle circumstance, calf circumstance, and handgrip strength (HGS) were measured. PA and appendicular skeletal muscle mass were examined by bioelectrical impedance analysis. According to the diagnostic criteria of the 2019 consensus of Asian Sarcopenia Working Group, the patients were divided into two groups: non-sarcopenia and sarcopenia. The study included 445 patients with a 22.2% prevalence of sarcopenia. RESULTS: PA was different between the non-sarcopenia and sarcopenia groups (5.02° versus 4.18°; P < 0.001). Pearson correlation showed that PA was related to diagnostic and confounding factors of sarcopenia. After adjusting for all potential confounding factors, multiple linear regression analysis showed diagnostic components of sarcopenia (HGS and skeletal muscle mass index [SMI]) could predict 25.3% of PA variation and logistic regression analysis showed PA (odds ratio, 0.309; P < 0.001) were related to sarcopenia. Then receiver operating characteristic curve showed the cutoff value of 4.25° with area under the curve of 0.785 for PA. CONCLUSIONS: PA is related to diagnostic components of sarcopenia, HGS and SMI. PA can be useful in the diagnosis of sarcopenia in older male patients with cancer. The cutoff value proposed in this study was 4.25°.

Serum tau levels are increased in patients with hyperthyroidism.

Hyperthyroidism may cause cognitive decline and increases the risk of Alzheimer's disease (AD), the major form of dementia; however, the underlying mechanism of this relationship is unclear. AD is associated with increased serum levels of tau. In this study, we investigated the relationship between serum thyroid hormones (THs) and tau. Fifty participants diagnosed with hyperthyroidism and fifty euthyroid counterparts were included and received clinical examinations. Serum concentrations of thyroid-stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3) and tau protein were assessed. The total tau protein level was significantly higher in hyperthyroidism participants than in their euthyroid counterparts. The level of circulating total tau had a significant positive association with the serum concentrations of FT3 and FT4. Total tau level was increased in the low TSH group and the serum THs decreased with the increase of age. These findings reveal that peripheral THs are associated with the serum concentration of tau, which may be involved in the pathogenesis of AD, suggesting a potential therapeutic target of AD via hyperthyroidism therapy.

The Effect of Type 2 Diabetes Mellitus on Neuropsychological Symptoms in Chinese Early Alzheimer's Disease Population.

OBJECTIVE: To explore the effect of Type 2 diabetes mellitus (T2DM) on the development of neuropsychiatric symptoms (NPS) in early Alzheimer's disease (AD). METHODS: From September 2017 to March 2019, a cross-sectional study was conducted on the clinical data of 158 early AD patients over 65 years old in the Department of Neurology of Daping Hospital. All early stage of AD patients were divided into early stage of AD with NPS group and early stage of AD without NPS group according to the presence or absence of NPS. Clinical data of age, sex, body mass index (BMI), smoking and alcohol consumption, history of hypertension, hyperlipidemia, white matter leisure (WML) and T2DM, MMSE, CDR and NPI-Q scores were collected. Multivariate logistic regression analyses were performed to examine the relationship between T2DM and NPS in early AD. RESULTS: Compared with the early stage of AD group without NPS, the early stage of AD group with NPS was older, the proportion of women was higher, the proportion of T2DM, hypertension, hyperlipidemia and WML was higher, and the MMSE score was lower (P< 0.05). T2DM was an independent risk factor for NPS in early stage of AD patients (OR 3.48, 95% CI 2.91-3.84). The incidence of T2DM in AD patients with depression, anxiety, nighttime behavioral disturbances, and appetite disturbances was significantly higher than in AD patients without these symptoms. T2DM was an independent risk factor of depression (OR 2.04, 95% CI 1.71-2.38), anxiety (OR 1.69, 95% CI 1.38-1.97), nighttime behavioral disturbances (OR 1.95, 95% CI 1.75-2.13) and appetite disturbances (OR 1.62, 95% CI 1.33-1.94) in early AD patients. CONCLUSION: T2DM was an important independent risk factor for NPS in early AD, which promotes the occurrence of depression, anxiety, nighttime behavioral disturbances and appetite disturbances in early AD.

Naturally Occurring Antibodies to Tau Exists in Human Blood and Are Not Changed in Alzheimer's Disease.

Hyperphosphorylated tau is an important pathological agent in Alzheimer's disease (AD). Tau effluxes from the brain to the blood could potentially stimulate the production of naturally occurring antibodies (NAbs). We aimed to investigate whether NAbs to tau (NAbs-tau) was generated in human blood and to figure out the alteration of plasma NAbs-tau level in AD patients. About 192 AD patients and 192 age-matched and non-demented controls (NC) were enrolled in the present study. Immunofluorescence staining and western blot assays were used to confirm the existence of NAbs-tau in human blood. The plasma level of NAbs-tau in NC and AD group was analyzed by ELISA. Immunofluorescence staining and western blot assays confirmed the existence of NAbs-tau in human blood. However, no significant difference in the plasma level of NAbs-tau was observed between NC and AD group. Furthermore, the plasma level of NAbs-tau had no significant correlation with MMSE scores. The present study confirmed that NAbs-tau exists in human blood but does not differ in level between the NC and AD group. Plasma NAbs-tau is not a reliable biomarker for AD.

Light alters the responses of two marine diatoms to increased warming.

In this study, we examined the effects of increased temperature (15, 20 and 25 °C) and different light levels (50, 200 µmol photons m-2 s-1) on two widely distributed diatoms, namely Phaeodactylum tricornutum and Thalassiosira weissflogii. Results showed that increasing light level counteracted the negative effects of high temperature on photosynthesis in both species, suggesting an antagonistic interaction between light and temperature. Contrary to the above results, light limitation diminished the temperature-sensitivity of carbonic anhydrase activity in two diatoms. We also observed species-specific responses of biomass, where increased temperature significantly decreased the biomass of P. tricornutum at both low and high light levels but showed no effects on T. weissflogii. Our study demonstrated that light can alter the physiological responses of diatoms to temperature but also revealed interspecific variations. We predict that in the future ocean with shallower upper mixed layer, T. weissflogii may be more competitive than P. tricornutum.

Time-Resolved Monitoring of Intracellular Processes with a Cyclical On-Off Photoswitchable Nanoprobe.

Fluorescent microscopic imaging with the help of small-molecule probes (chemoprobes) is one of the most feasible approaches for noninvasive sensing of intracellular molecules. However, the "always on" property of current chemoprobes failed to achieve time-resolved monitoring. Here, we report the development of a supramolecular nanoassembling strategy to integrate multiple functions on one nanoscale probe (nanoprobe) with a cyclical on-off switchable sensing ability. The reversal of the nanoprobe can be rapidly achieved by converting the single-wavelength near-infrared (NIR) laser to two-way emissions by a lanthanum nanoparticle core that is sensitive to the light power density. Through regulating the NIR power density, the azobenzene derivative, which was doped in the surface of the lipid bilayer of the nanoprobe, can act as an "impeller" and "brake" for bio-benign activation and deactivation, respectively, of the nanoprobe in biological applications. A reduced nicotinamide adenine dinucleotide nanoprobe was constructed as the model to demonstrate precise and time-resolved monitoring of intracellular processes including cancerous glycolysis and ligand-induced enzymatic processes. We envision that this cyclical on-off switchable nanoprobe strategy will hold great promise for endowing universal chemoprobes with high precision and temporal resolution.

Red-Near-Infrared Fluorescent Probe for Time-Resolved in Vivo Alkaline Phosphatase Detection with the Assistance of a Photoresponsive Nanocontainer.

The monitoring of alkaline phosphatase (ALP) activity in different tissues is significant for disease diagnosis and therapy. However, the time-resolved in vivo sensing of ALP activity remained unresolved. Herein, a novel red-near-infrared fluorescent ALP probe (Cl2-BDCM-ALP) based on a dichloro-substituted dicyanomethylene-4H-chromene derivative was designed and synthesized with high fluorescence efficiency and stability under biological pH range. By using Cl2-BDCM-ALP, ALP activity under an acidic microenvironment such as a tumor site can be sensitively imaged, which cannot be achieved by some previously reported ALP probes. By further loading the Cl2-BDCM-ALP into a near-infrared (NIR) light-responsive nanocontainer, time-resolved long-term imaging of ALP activity was facilely achieved with noninvasive NIR light remote control. Time-resolved variation of ALP activity of the drug-induced acute liver injury mice was successfully monitored in vivo for the first time. This strategy holds great promise in the in situ ALP detection under a broad pH range with temporal resolution.

Efficacy and safety of NAD+ ADP-ribosyltransferase 1 agonist versus Donepezil in elderly Chinese patients with Alzheimer disease: A novel target for effective therapy.

This pilot study designed to evaluate the efficacy and safety of NAD+ ADP-ribosyl transferase 1 (NART) agonist in comparison with Donepezil (DNP) in elderly Chinese patients with Alzheimer disease (AD). In the present clinical trial, Chinese elderly patients aged >65 years with a confirmed diagnosis of AD were enrolled. The patients received NART agonist (test, DAG-structured PKC blockers (GF109203X)) or DNP 10mg daily (reference) for 6 months. The efficacy and safety data were collected from 120 patients (60 patients in each group) every 3 weeks until 6 months. The primary endpoints were to assess the change in cognitive score from baseline in both the treatment group. The result of the present study showed that the patients treated with DNP and NART agonist have similar efficacy and safety profile. Considering the clinical benefit, improvement in sign and symptoms of was numerically greater in DNP-treated patients as compared to NART agonist. However, a statistical difference in terms of clinical benefit was similar between both the treatment groups. Overall, both the study drugs were found comparable in relieving the symptoms of AD. This indicates that NART is a potential target for the treatment of AD in China. The results of the present study may help to design a large clinical trial to evaluate the efficacy and safety of NART agonist in comparison with DNP in AD patients.

An array consisting of glycosylated quantum dots conjugated to MoS2 nanosheets for fluorometric identification and quantitation of lectins and bacteria.

A fluorescent array based on the use of saccharide-functionalized multicolored quantum dots (s-QDs) and of 4-mercaptophenylboronic acid-functionalized MoS2 nanosheets (PBA-MoS2) was constructed for multiple identification and quantitation of lectins and bacteria. In this array, the fluorescence of the s-QDs is quenched by the PBA-MoS2 nanosheets. In the presence of multiple lectins, s-QDs differentially detach from the surface of PBA-MoS2 nanosheets, producing distinct fluorescence response patterns due to both quenching and enhancement of fluorescence. By analyzing the fluorescence responses with linear discriminant analysis, multiple lectins and bacteria were accurately identified with 100% accuracy. The limits of detection of Concanavalin A, Pisum sativum agglutinin, Peanut agglutinin, and Ricius communis I agglutinin are as low as 3.7, 8.3, 4.2 and 3.9 nM, respectively. The array has further been evidenced to be potent for distinguishing and quantifying different bacterial species by recognizing their surface lectins. The detection limits of Escherichia coli and Enterococcus faecium are 87 and 66 cfu mL-1, respectively. Graphical abstract Schematic of a fluorometric array based on the use of saccharides-functionalized quantum dots (s-QDs) and 4-mercaptophenylboronic acid-functionalized MoS2 (PBA- MoS2) nanosheets. This array was successfully applied to simultaneously analysis of lectins, bacteria in real samples with high sensitivity and accuracy.

Inflammatory cytokines and cells are potential markers for patients with cerebral apoplexy in intensive care unit.

Cerebral apoplexy is a disease caused by obstruction of the blood circulation in the brain. Evidence has indicated that inflammatory cytokines are implicated in ischaemic cerebral apoplexy and are regarded as a general cardiovascular risk factor, which may be a possible immediate trigger, a component of the response to tissue injury and a therapeutic target. The present study investigated changes of inflammatory cytokines and cells in patients with cerebral apoplexy at the intensive care unit (ICU). The plasma concentrations of inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-4, IL-6, IL-8, IL-10, IL-1ß and IL-17A were evaluated using ELISA. Changes in the plasma concentrations of inflammatory cells were detected by using flow cytometry. The results indicated that serum levels of TNF-α, IL-4, IL-8, IL-1ß and IL-17A were upregulated in patients with cerebral apoplexy compared with those in healthy individuals, while those of IL-6 and IL-10 were downregulated. Furthermore, it was demonstrated that the plasma concentration of lymphocytes, granulocytes and mononuclear cells was decreased in patients with cerebral apoplexy in the ICU compared with that in healthy individuals. Of note, humoral as well as cellular inflammatory cytokines were evidently increased in patients with cerebral apoplexy in ICU. In conclusion, the present study provided evidence that inflammatory cytokines and inflammatory cells are upregulated, while anti-inflammatory cytokines are downregulated in patients with cerebral apoplexy in an ICU setting. These results suggest that anti-inflammatory interventions may be beneficial either in the prevention or acute treatment of patients with cerebral apoplexy.

Simultaneous quadruple-channel optical transduction of a nanosensor for multiplexed qualitative and quantitative analysis of lectins.

A multichannel optical nanosensor capable of identifying and quantitating multiple lectins simultaneously was developed. The quadruple channel of fluorescence and scattering signals can be in situ collected from the same solution system, which offers high accuracy, discrimination resolution and measurement convenience. This nanosensor can in principle be generalized to the analysis of all lectins and saccharide binding organisms.