Adipose-derived stem cell (ADSC) transplantation has emerged as a potential tool for the treatment of cardiovascular disease and skin wounds. However, with a limited renewal capacity and the need for mass cells during the engraftment, strategies are needed to enhance ADSC proliferative capacity. In this study, we explored the effects of Exendin-4, a glucagon-like peptide-1 analog, on the growth of ADSCs, focusing in particular on phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt) and Wnt signaling pathways. Firstly, ADSCs were isolated and cultured in vitro. Then, flow cytometry demonstrated that ADSCs were positive for CD44, CD90 and CD29 but negative for CD31, CD34, and CD45. Exendin-4 (0-200 nM) treatment increased ADSC proliferation. In order to examine specific signaling pathways, a western blotting assay was performed. Our results demonstrate that after treated with 50 nM Exendin-4 for 48 h, the phosphorylation of PI3K, Akt, and GSK3 beta were increased and phosphorylation of beta-catenin was decreased. From these results, we concluded that PI3K-Akt and Wnt-beta-catenin signaling pathways mediate Exendin-4 induced ADSC proliferation, the function of which might contribute to the regulation of ADSC proliferation. Our findings provided new insights into the function of the mechanisms underlying Exendin-4 of ADSCs.
LRRK2 is the most common genetic cause of PD. G2385R and R1628 P variants are the most common variants of LRRK2 in Chinese populations. Consensus on the clinical features of G2385R and R1628 P related PD has not been reached yet, although it had been widely studied. In our study, genotype analyses were conducted on 721 PD patients of Chinese origin. A total of 62 G2385R carriers, 32 R1628 P carriers and 623 idiopathic PD patients underwent the following clinical feature analysis. Motor symptoms, non-motor symptoms and co-morbidities were the targeted features to be analyzed. As a result, Neither the G2385R nor the R1628 P carriers showed significant clinical feature differences when compared to the idiopathic PD patients, so did the comparison between the G2385R and the R1628 P carriers. In conclusion, the clinical features of PD patients with LRRK2 G2385R or R1628 P variants were similar to those of idiopathic PD.
An immediate feedback after action facilitated reinforcement learning in dynamically varying environments. With several seconds delay, a series of event-related potential (ERP) studies have recently conducted to explore how delayed feedback influences learning processes and corresponding brain activities by measuring the Reward Positivity and N170 component. However, it remains unclear how does our brain process a feedback that is delayed longer and interrupted by other trials. In the present study, participants were asked to undertake a time estimation task in two different conditions. Feedback was presented right after their actions in the immediate feedback condition, while it was presented after another five trials in the delayed feedback condition. By recording feedback related activities, we aim to test whether, or not, delayed feedback impairs reinforcement learning, the Reward Positivity and N170 amplitude. The behavioural results show that delayed feedback can reduce behavioural adjustment efficiency from trial-to-trial. To reduce component overlapping, we adopted the temporospatial principal components analysis (PCA) to separate the Reward Positivity from other ERP components. Results indicate that the Reward Positivity is decreased in the delayed feedback condition compared to the immediate feedback condition, however, no difference of N170 amplitude is found between the two conditions. These results indicate that delayed feedback impairs reinforcement learning process in terms of behavioural adjustment and brain activities even though these feedbacks are truly associated with participants' previous actions.
Severe hemorrhagic shock induces cognitive dysfunction by promoting cell death mediated by activating endoplasmic reticulum (ER) stress. Sevoflurane postconditioning prevents neuronal apoptosis against cerebral ischemia/reperfusion injury. It is unknown if this protective effect on hemorrhagic shock and resuscitation rats (HSR) is associated with ER stress attenuation. Male adult Sprague-Dawley rats were subjected HSR by removing 40% blood volume within 30 min, and 60 min later the animals were resuscitated with infusion of the removing blood in 30 min. Sevoflurane postconditioning was performed by inhaling sevoflurane at three different concentrations (0.5, 1.0, 1.5 MAC) at the onset of resuscitation for 30 min. Severe hypotension (mean arterial pressure 40-45 mmHg) occurred in the shock session for 60 min accompanying with significantly elevated lactate, decreased BE and pH values in arterial blood gas analysis. There were impaired spatial learning and memory following HSR indicated by persistently longer escape latency and lower correct rate, as well as less duration and crossing in the target quadrant by using Morris water maze and Y-maze tests. In the hippocampal CA1 region, there was significantly higher activity of caspase-3 induced by HSR. HSR also elevated the expression of inositol-requiring enzyme 1 alpha (IRE1 alpha) and caspase-12 in the hippocampus by western blot analysis. Sevoflurane postconditioning at 1.0 and 1.5 MAC significantly reversed these changes. These findings suggested that sevoflurane postconditioning could improve spatial learning and memory deficits induced by severe hemorrhagic shock and subsequent resuscitation. The suppression of endoplasmic reticulum stress provided critical contribution in neural apoptosis mediated by IRE1 alpha-caspase-12 pathway.
Amisulpride is an effective antipsychotic for the treatment of schizophrenia with a lower propensity for extra pyramidal adverse effects than conventional antipsychotics. However, no study has investigated white matter (WM) integrity in patients with schizophrenia in relation to treatment response after amisulpride administration. Here, we investigated the associations of WM integrity with severity reductions in clinical symptoms in drug-free patients with schizophrenia at an early stage of amisulpride treatment. Nineteen patients with schizophrenia (SZ) and 15 healthy controls (HCs) participated in the present study. Diffusion tensor imaging data were acquired from all participants at baseline. All SZ participants began treatment with 200 mg of amisulpride per day. The dose was increased up to 1200 mg/day within 2 weeks depending on the severity of clinical symptoms, and maintained for the subsequent 6 weeks. Initially, and after 8 weeks of amisulpride treatment, SZ participants were assessed for the severity of overall illness, positive and negative symptoms, and motor side effects. SZ participants showed lower integrity in several WM regions, including the corpus callosum and fronto-temporal connections, when compared to HCs. Furthermore, lower WM integrity in fronto-temporo-limbic regions at baseline was found to be associated with severity reductions in positive symptoms after 8 weeks. Our findings suggest that WM integrity at the early stage of treatment may serve as a possible predictive marker for treatment response.
The underlying mechanism of apolipoprotein E epsilon 4 (APOE epsilon 4) in the pathogenesis of Alzheimer's disease (AD) remains elusive. We hypothesize that synaptic function is differentially affected by APOE isoforms. Levels of CSF SNAP-25 were compared between APOE epsilon 4 carriers and noncarriers in 55 participants with normal cognition, 75 patients with mild cognitive impairment (MCI), and 16 patients with mild AD dementia. We investigated relationships between SNAP-25 levels and age, gender, education, CSF A beta 42, and tau protein. We found that levels of SNAP-25 in CSF were substantially greater in APOE epsilon 4 carriers compared to noncarriers with MCI. There was no significant difference in SNAP-25 levels between APOE epsilon 4 carriers and noncarriers with normal cognition or AD. CSF SNAP-25 levels were associated with MMSE and CSF A beta and tau levels. In summary, APOE epsilon 4 may affect CSF SNAP levels in MCI patients, suggesting an important role of APOE epsilon 4 in synaptic dysfunction leading to AD.
Intellectual disability (ID) is one of the most prevalent chronic developmental brain disorders or phenotype of syndromic ID, affecting nearly 1-2% of the general population worldwide. Over recent decades, tremendous effort and high-throughput platforms have been devised to explore the complex heterogeneity, numerous genes and variants have been associated with the ID, especially de novo mutations and copy number variants. An organized resource containing the increasing genetic data is imperative to assist ID research. In this study, the integrative and annotated intellectual disability database has been developed, named 'IDGenetics', which contains known information about ID, including 815 genes and 17102 variants associated with 918 clinical diseases (3001 clinical phenotype) collected from 3822 publications and ID-related databases. Furthermore, in-depth data mining was performed to obtain an understanding of each entry, including functional annotation, gene/disease/phenotype network establishment and overlap analysis focusing on comorbidity. 1478 candidate genes (483 high-confidence and 995 low-confidence) were collected and prioritized by adopting the annotations of 12 functional prediction tools and algorithm. In addition, IDGenetics database provides concise search methods, convenient browsing functions, intuitive graphical displays and constantly updated features. IDGenetics will be a valuable and integrative resource for deciphering the genetic and functional architecture of ID and the improvement of clinical diagnosis, intervention and treatment.
Prodynorphin (PDYN) binds to k-opioid receptors (KOPr; encoded by OPRK1) and is known to regulate dopaminergic tone, making this system important for drugs addiction. Dynorphin (Dyn)/KORr system are powerful effectors of stress-induced alterations in reward processing and dysphoric states. Thus, We identified 11 potential functional SNPs and one variable number of tandem repeat (VNTR) in this system, performed a case-control association analysis, investigated particular disease phenotypes, assessed the joint effect of variants in two genes, carried out a meta-analysis to analyze the association between this VNTR and Heroin dependence (HD) risk. Eleven single-nucleotide polymorphisms (SNPs) were genotyped using SNaPshot SNP technology. Participants included 566 healthy controls and 541 patients with HD. We found that PDYN polymorphisms modulate the susceptibility to HD. An increased risk of HD was significantly associated with H alleles of PDYN VNTR (chi(2) = 10.824, p = 0.001, OR = 1.419, 95% CI = 1.151-1.748). In addition, the results revealed the patients with the HH genotype showed greater number of withdrawal instances (F(2538) = 7.987, p = 0.0004) compared to the patients with the LL genotype. The Meta-analysis showed the pooled effect of the H allele at this locus is a risk factor for HD in Chinese Han. Gene-gene interaction analysis indicated strong interactions between PDYN rs3830064, 68-bp VNTR and OPRK1 rs16918842, rs3802279. These findings support the important role of PDYN polymorphism in HD, and may guide future studies to identify genetic risk factors for HD.
Patients with spinal cord injury (SCI) have an increased risk for developing type 2 diabetes. It is unknown whether the pancreatic-islet microvascular vasomotion is involved. We used female C57BL/6 mice and a 100-kilodyne T10 Infinite Horizons contusion SCI (or T10 laminectomy) to detect blood glucose and pancreatic-islet microvascular vasomotion. Blood glucose obtained from tail vein was detected using one Touch UltraEasy glucometer. Glucose tolerance test was performed by n-glucose administration intraperitoneally. Functional status of pancreatic-islet microvascular vasomotion was determined by laser Doppler monitoring. Expressions of insulin and glucagon were determined by immunohistochemistry. Expression of VEGF-A was determined by immunohistochemistry and Western blotting. Our result demonstrated that blood glucose was significantly increased at 4 h postinjury compared to that in sham group, with continuous higher blood glucose until 4 days postinjury (p < 0.05). SCI mice at day 7 and day 14 had significantly impaired glucose tolerance following glucose administration (p < 0.01). Average blood perfusion, amplitude, frequency, and relative velocity of vasomotion were significantly lower at 6 h postinjury than those in the sham group (p < 0.05), which were gradually upregulated over time. The expression of insulin was decreased, while the expression of glucagon was increased at 6h postinjury. Similarly, the expression of VEGF-A was significantly decreased at 6h postinjury, compared to that in sham group (p < 0.05), with slight increases by 14 days postinjury. Our study suggests that the functional status of pancreatic-islet microvascular vasomotion is impaired after injury, which may have implications for developing effective therapeutic interventions for SCI.
Amyloid-beta proteins deposition and aggregation occur in extracellular space and form neuritic plaques in Alzheimer's disease (AD) brain. Beta-site amyloid precursor protein cleaving enzyme 1 (BACE1)/ beta-secretase and gamma-secretase Presenilin 1 (PSEN1) conduct sequential cleavage of amyloid- beta precursor protein (APP) and yield amyloid- beta proteins. However the details of the interactions of APP with the enzymes and transportation of catalytic products are unclear. Here we reveal distinctive targeting patterns of the proteins in subcellular organelles in N2A cells. We find all three proteins co-localize in endosomes with APP and PSEN1 co-localize and associate on cell membrane and nucleus. By selectively knocking down BACE1 or PSEN 1 with siRNA, we discover that BACE1 functions as the enzyme initiating the first cleavage step and serves a scaffold for APP and PSEN1 endocytosis. PSEN1 knocking-down only leads to the reduction of BACE1 in cell membrane and nucleus. We conclude that BACE1 facilitates the transportation of APP and formation of the complex with gamma-secretase, resulting in the stepwise cleavages of APP. After BACE1 cleavage APP binds to PSEN1 and transfers to cell membrane or nucleus for final processing and amyloid genesis.
Background: Development of Parkinson's disease (PD) is attributed to both genetic and environmental factors. Furthermore,GAPDH may play a key role in the development of neurodegenerative disease. Examination of genetic polymorphism in patients with sporadic PD will help uncover the mechanisms of PD pathogenesis and provide new insights into the treatment of PD. Methods and results: The SNaPshot method was applied to determine the gene sequences in 265 patients with idiopathic PD and 269 control cases (sex- and age-matched). The rs1136666 polymorphism of GAPDH was determined to be closely associated with PD. Subsequently, the CC genotype of the rs1136666 fragment was transfected into SH-SY5Y cells via a plasmid. The genetic expression of rs1136666 CC could induce SH-SY5Y cell injury and apoptosis via regulation of the oxidant-antioxidant and apoptosis antiapoptosis balance. rs1136666 CC of the GAPDH had a pro-apoptotic effect similar to that of rotenone, and combination of the rs1136666 CC genetic variation and the rotenone neurotoxic effect could aggravate oxidative stress, cell injury, and apoptosis better than either single treatment alone. Conclusion: This study confirmed that the rs1136666 CC allele of theGAPDH increased the risk of PD, particularly in older male patients.
Neurofilament light chain (NFL) in cerebrospinal fluid (CSF) is a promising biomarker candidate which may discriminate atypical parkinsonian disorders (APD), mainly including multiple system atrophy (MSA), progressive supranuclear palsy (PSP), and corticobasal degeneration (CBD), from Parkinson's disease (PD). We aim to evaluate the diagnostic accuracy of CSF NFL level as a differentiating biomarker between APD and PD. Databases of PubMed, OVID and Web of Science were searched for studies (published until May 31, 2017) that reported on CSF NFL as a diagnostic biomarker between APD and PD. Eight studies were pooled in this meta-analysis, including 341 PD and 396 APD patients and 388 healthy controls. The pooled sensitivity was 82% (95% CI, 68%-91%) and specificity was 85% (95% CI, 79%-89%) in differentiating APD from PD. The pooled positive likelihood ratio (PLR), negative likelihood ratio (NLR) and diagnostic odds ratio (DOR) were 5.4 (95% CI, 3.6%-8.1%), 0.21 (95% CI, 0.11%-0.40%), and 25 (95% CI, 9%-69%) respectively; and the area under the curve (AUC) was 0.89 (95% CI, 0.86%-0.91%). Subgroup analysis revealed sensitivity and specificity were significantly influenced by study design. The APD subtypes, disease duration and severity were the main heterogeneity sources in specificity. The results of Deeks' test revealed a low risk of publication bias. The CSF NFL level may be used as a biomarker in discriminating APD from PD with high diagnostic accuracy at an early stage of disease. Large and longitudinal studies are still needed on individuals who are suspected to have APD.
Previous brain morphology-related diagnostic models for attention-deficit hyperactivity disorder (ADHD) were based on regional features. However, building a model of individual interregional morphological connectivity is a challenging task. This study aimed to identify children with ADHD utilizing a novel interregional morphological connectivity model and discover the discriminative patterns in patients. Therefore, novel interregional morphological patterns rather than regional patterns were extracted via surface-based analysis. The interregional morphological features were trained and tested using a hybrid machine learning method, which was implemented using the leave-one-out cross-validation (LOOCV) method to produce the optimized discriminative model and discriminative patterns. The inclusion of interregional morphological connectivity significantly improved the performance of the diagnostic models compared to the performance of the model constructed using regional features. The optimized discriminative model exhibited a total accuracy of 74.65%, a sensitivity of 75% and a specificity of 74.29%. The brain regions displaying altered morphological connectivity included the insula, the caudal anterior cingulate cortex, the frontal pole, and the postcentral cortex, among others. In addition, the altered connections correlated with the clinical symptoms. In summary, patients with ADHD exhibited altered morphological connectivity, which might be a potential biomarker for the classification of ADHD. The discriminative features will potentially benefit studies investigating the brain network mechanisms of ADHD.
Background: A number of genetic variants have previously been identified and associated with the risk of Alzheimer's disease (AD), including rs10838725 in CELF1, rs28834970 in PTK2B, rs17125944 in FERMT2, and rs10410544 in SIRT2 based on genome-wide association studies. Considering the overlap between the clinical manifestation and pathological characteristics of AD and Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS) and multiple system atrophy (MSA), we conducted a large sample study to investigate the associations between these variants and these three common neurodegenerative diseases in a Chinese population. Methods: A total of 2449 patients, including 1219 PD, 870 sporadic ALS, and 360 MSA, and 821 healthy controls were examined for this study. All cases were genotyped for single-nucleotide polymorphisms using Sequenom iPLEX assay technology. Results: No significant differences were found in genotype distribution and minor allele frequencies between the four candidate variants and the three neurodegenerative diseases. However, a significant difference was found in the minor allele frequency of rs28834970 in PTK2B between PD patients with normal and abnormal cognitive function (p = 0.001). Moreover, the minor allele "C" was associated with an increased risk for cognitive impairment in PD (OR = 1.84). Although this observation was not significant (p = 0.064), the mean Addenbrooke's Cognitive Examination-Revised (ACER) score of PD patients with the risk allele of rs28834970 was 2.913 +/- 1.569 points lower than that of PD patients without the risk allele. Conclusion: This study provides new insight into some of the phenotypes that may share the common pathogenesis of different neurodegenerative diseases.
Alcohol abuse and anxiety disorders often occur concurrently, but their underlying cellular mechanisms remain unclear. N-methyl-D-aspartic acid receptors (NMDARs) have recently received attention from those interested in the neurobiology of anxiety. A chronic alcohol exposure rat model (28 consecutive days of 20% alcohol intake and 6 h of withdrawal) was established. Here, we investigated the NMDAR1 (NR1), Ca2+/calmodulin-dependent protein kinase II (CaMKII) and extracellular signal-regulated kinases (ERK) pathway in the modulation of anxiety-like behaviors in rats exposed to an open field and elevated plus maze (EPM) through systematic injections of memantine (a NMDAR inhibitor). We found that the NR1-CaMKII-ERK signaling pathway was activated after alcohol withdrawal in medial prefrontal cortex (mPFC) and nucleus accumbens shell (NAcSh) but not core (NAcC). Memantine treatment greatly ameliorated anxiety-like behavior in the rats experiencing alcohol withdrawal. Moreover, memantine uniformly suppressed the phosphorylation of NR1-CaMKII-ERK pathway induced by alcohol withdrawal. Our results suggest that activation of the NR1-CaMKII-ERK pathway in the mPFC and NAcSh is an important contributor to the molecular mechanisms underlying alcohol withdrawal-induced anxiety behaviors. NMDAR signaling pathway inhibitors are thus potential therapeutics for treating alcohol abuse.