Multi-technology integrated network pharmacology-based study on phytochemicals, active metabolites, and molecular mechanism of Psoraleae Fructus to promote melanogenesis.

ETHNOPHARMACOLOGICAL RELEVANCE: According to the Compendium of Materia Medica (Shizhen Li, Ming dynasty) and Welfare Pharmacy (Song dynasty), Psoraleae Fructus (PF), a traditional Chinese medicine (TCM) has a bitter taste and warm nature, which has the effect of treating spleen and kidney deficiency and skin disease. Although PF has been widely used since ancient times and has shown satisfactory efficacy in treating vitiligo, the active substances and the mechanism of PF in promoting melanogenesis remain unclear. AIM OF THE STUDY: To explore the active substances and action mechanisms of PF in promoting melanogenesis. MATERIALS AND METHODS: Firstly, UPLC-UV-Q-TOF/MS was used to characterize the components in PF extract and identify the absorption components and metabolites of PF after oral administration at usual doses in rats. Secondly, the active substances and related targets and pathways were predicted by network pharmacology and molecular docking. Finally, pharmacodynamic and molecular biology experiments were used to verify the prediction results. RESULTS: The experimental results showed that 15 compounds were identified in PF extract, and 44 compounds, consisting of 8 prototype components and 36 metabolites (including isomers) were identified in rats' plasma. Promising action targets (MAPK1, MAPK8, MAPK14) and signaling pathways (MAPK signaling pathway) were screened and refined to elucidate the mechanism of PF against vitiligo based on network pharmacology. Bergaptol and xanthotol (the main metabolites of PF), psoralen (prototype drug), and PF extract significantly increased melanin production in zebrafish embryos. Furthermore, bergaptol could promote the pigmentation of zebrafish embryos more than psoralen and PF extract. Bergaptol significantly increased the protein expression levels of p-P38 and decreased ERK phosphorylation in B16F10 cells, which was also supported by the corresponding inhibitor/activator combination study. Moreover, bergaptol increased the mRNA expression levels of the downstream microphthalmia-associated transcription factor (MITF) and tyrosinase in B16F10 cells. Our data elucidate that bergaptol may promote melanogenesis by regulating the p-P38 and p-ERK signaling pathway. CONCLUSIONS: This study will lay a foundation for discovering potential new drugs for treating vitiligo and provide feasible ideas for exploring the mechanism of traditional Chinese medicine.

Effect of PLA2G6 and SMPD1 Variants on the Lipid Metabolism in the Cerebrospinal Fluid of Patients with Parkinson's Disease: A Non-targeted Lipidomics Study.

INTRODUCTION: Sleep patterns are more frequently interrupted in patients with Parkinson's disease (PD), and it is still unclear whether genetic factors are involved in PD-related sleep disorders. In this study, we hypothesize that PD-associated genetic risk affects lipid metabolism, which in turn contributes to different types of sleep disorders. METHODS: We used a non-targeted lipidomics to explore the lipid composition of cerebrospinal fluid (CSF) exosomes derived from patients with PD carrying phospholipase A2 Group VI (PLA2G6) and sphingomyelin phosphodiesterase 1 (SMPD1) mutations. RESULTS: PLA2G6 mutations (c.1966C > G, Leu656Val; c.2077C > G, Leu693Val; c.1791delC, His597fx69) significantly increase the exosomal content of glycerophospholipids and lysophospholipids, specifically phosphatidylcholine (PC) and lysophosphatidylcholine (LPC). Exosome surface presence of melatomin receptor 1A (MTNR1A) was detectable only in patients with PLA2G6 mutations. We have further shown that, in patients with PD carrying PLA2G6 mutations, sleep latency was significantly longer compared to those carrying WT PLA2G6, and we speculate that functional PLA2G6 mutations lead to structural changes and lipid deregulation of exosomes, which in turn alters exosomal cargo and affects PD-related sleep disorders. In SMPD1, G508R variant-carrying patients with PD abundance of sphingomyelins was significantly higher and had significantly shorter rapid eye movement sleep. CONCLUSIONS: Our study demonstrated that the disturbed composition and function of CSF-derived exosome lipidome during the pathological stage of PD may affect different types of sleep disorder in PD.

Exosomes rich in Wnt5 improved circadian rhythm dysfunction via enhanced PPARγ activity in the 6-hydroxydopamine model of Parkinson's disease.

Sleep disorder is one of the most common non-motor symptoms in Parkinson's disease (PD) and even appear as early symptoms. Here we investigated the therapeutic potential of mesenchymal stem cell-derived exosomes (MSC-EXOs) on sleep disorder in PD rats. 6-hydroxydopa (6-OHDA) was used to establish the PD rat model. BMSCquiescent-EXO and BMSCinduced-EXO groups were given intravenous injection 100 µg/g per day for 4 weeks, while control groups were given intravenous injection of the same volume of normal saline. The total sleep time, slow-wave sleep time and fast-wave sleep time in the BMSCquiescent-EXO and BMSCinduced-EXO groups were significantly prolonged (P < 0.05) compared with PD group, while the awakening time was significantly shortened (P < 0.05). In addition, increased levels of dopamine (P < 0.05) and 5-hydroxytryptamine (P < 0.05) levels were observed in the striatum of BMSCquiescent-EXO and BMSCinduced-EXO groups. Further, qPCR and western blot revealed that the mRNA levels of CLOCK, BMAL1 and PER2 in suprachiasmatic nucleus (SCN) were notably increased in BMSCquiescent-EXO and BMSCinduced-EXO groups compared to those from PD rats. More importantly, peroxisome proliferation-activated receptor γ (PPARγ) activities were significantly enhanced after treatment with BMSCquiescent-EXO and BMSCinduced-EXO. JC-1 fluorescence staining showed that mitochondrial membrane potential imbalance was repaired after inoculation of BMSCinduced-EXO. In summary, MSC-EXOs showed the improvement of sleep disorder in PD rats through recovering circadian rhythm associated gene expression. The potential mechanisms may be related with increased PPARγ activities and rescued mitochondrial membrane potential imbalance in Parkinson striatum.

Optic Nerve Sheath Fenestration for Progressive Visual Loss in Cerebral Venous Sinus Thrombosis: A Long-Term Retrospective Observational Study.

INTRODUCTION: Progressive cerebral venous sinus thrombosis (CVST)-induced visual loss remains problematic, despite decreasing overall mortality owing to early diagnosis and aggressive treatment. Optic nerve sheath fenestration (ONSF) improves or stabilizes visual function in patients with idiopathic intracranial hypertension; however, its role in CVST awaits elucidation. We evaluated the efficacy and safety of ONSF in resolving CVST-induced visual impairment based on long-term observation. METHODS: This observational study included 18 patients with progressive CVST-induced visual loss, who had undergone ONSF between 2012 and 2021. Patients received maximum medical therapy, including anticoagulants and intracranial pressure (ICP)-lowering medications. The best-corrected visual acuity (BCVA), visual fields (VFs), and optic nerve head were assessed at baseline, at 1 week after ONSF, and over 6 months after ONSF. Activities of daily living (ADL) and National Eye Institute Visual Function Questionnaire-25 (VFQ-25) scores were assessed at final follow-up. RESULTS: Thirty-one ONSF-treated eyes of 18 patients were included. The mean follow-up duration was 35.6 months (range 1 week-8 years). Two patients were lost to follow-up. Before ONSF, all patients were still experiencing progressive visual loss despite receiving adequate anticoagulation and ICP-lowering therapy. Postoperative BCVA remained stable or improved in 25/31 eyes (80.6%) 1 week postoperatively and 17/28 eyes (60.7%) upon final follow-up. All papilledema resolved postoperatively. No complications were reported except for one transient postoperative diplopia. The median ADL score was 100 (range 25-100), and the mean total VFQ-25 score was 40.6 (range 9.5-87.3). CONCLUSION: This was the largest study to describe ONSF's role in CVST based on a long-term follow-up. Considering its efficacy and favorable safety, ONSF can be considered an important adjunctive approach to resolving progressive visual loss of CVST patients, on the basis of anticoagulation and ICP-lowering therapy.

Cerebral venous sinus thrombosis (CVST) is a cerebrovascular disease that generally affects young patients. Medical treatments include anticoagulants, intracranial pressure (ICP)-lowering medications, and repeated lumbar punctures, effectively reducing CVST's mortality rate. However, CVST still carries a potential risk of progressive vision loss. Optic nerve sheath fenestration (ONSF) has been reported to be effective and safe in protecting visual function of patients with idiopathic intracranial hypertension. However, its efficacy and safety have not been evaluated in visual loss caused by CVST. We were the first to evaluate the efficacy and safety of ONSF in CVST-induced progressive visual loss based on long-term follow-ups. Before ONSF, all patients were still experiencing progressive visual loss despite receiving adequate anticoagulation and ICP-lowering therapy. We found ONSF to be 80.6% (1 week postoperatively) and 60.7% (after long-term follow-up of over 6 months) effective in stabilizing and/or improving visual function as well as 100% effective in papilledema resolution. Moreover, ONSF exhibited a favorable safety profile, with an extremely low complication rate of 5.6% despite under perioperative anticoagulation. Although visual impairment in CVST was reported to be uncommon, it often significantly affects quality of life and social value of patients. Thus, visual loss in CVST deserves more attention from neurologists, neurosurgeons, and ophthalmologists. Considering its efficacy and favorable safety, ONSF could be regarded a potentially important adjunctive approach to resolving progressive visual loss in CVST patients, on the basis of anticoagulation and ICP-lowering therapy.Procedural videos available for this article.

Mesenchymal stem cell-derived exosomes altered neuron cholesterol metabolism via Wnt5a-LRP1 axis and alleviated cognitive impairment in a progressive Parkinson's disease model.

Parkinson's disease (PD) is associated with abnormal metabolism of brain cholesterol, and the metabolites of neuronal cholesterol may also affect neurodegenerative progression. In this study, we aim to explore the therapeutic effect of BMSC derived exosomes on motor and cognitive deficits in α-synuclein (α-Syn) A53T transgenic mice, a progressive PD animal model. Results revealed that rotating rod performance of α-Syn A53T TG mice decreased by 45.4 %±8.6 % at the age of 12 months compared with wide-type (WT) mice. Striatum injection of BMSC quiescent exosomes (BMSCquiescent-EXO) and BMSC induced exosomes (BMSCinduced-EXO) rescued the rotation behavior (BMSCquiescent-EXO: 92.3 %±12.5 % P = 0.008; BMSCinduced-EXO: 102.3 %±16.7 %, P = 0.006). Although there was no difference in the escape latency within 5 days of Morris water maze learning between groups in the 12-month old mice. The exploration latency was shorter (p < 0.05) in BMSCquiescent-EXO and BMSCinduced-EXO groups, the number of explorations and novel object recognition index were significantly increased (p < 0.05). More importantly, the total cholesterol level was increased (p < 0.05), while the content of 24S-hydroxycholesterol significantly decreased (p < 0.05) after intrastriatal injection BMSCquiescent-EXO and BMSCinduced-EXO in A53T group. Liquid chromatography-mass spectrometry (LC/MS) was performed to profile phospholipid metabolites in lipid raft of hippocampal neurons, demonstrating that BMSCquiescent-EXO injection caused the decreasing relative percentages of phosphatidylglycerol (PG) and phosphatidylethanolamine (PE) compared to those in A53T mice, while the relative percentages of phosphatidylinositol (PI), phosphatidylserine (PS), and phosphatidylcholine (PC) increased. The cholesterol content of lipid rafts was lower in BMSCquiescent-EXO and BMSCinduced-EXO groups than that in A53T group (P < 0.05). In summary, exosomes isolated during BMSC dopaminergic neuron differentiation can significantly improve the motor, learning and memory ability of the progressive PD mice model, and its mechanism may be related to the change of altered phospholipid composition and cholesterol metabolism in hippocampal neurons.

Cerebrospinal Fluid Extracellular Vesicles with Distinct Properties in Autoimmune Encephalitis and Herpes Simplex Encephalitis.

Encephalitis mediated by autoantibodies against neuronal antigens and herpes simplex encephalitis (HSE) are seemingly separate causes of encephalopathy in adults. Autoimmune encephalitis (AE) is autoimmune in origin, and herpes simplex encephalitis is infectious. The purpose of this study was to examine the role of cerebrospinal fluid (CSF) exosomes from patients with antibody-positive AE and HSE. Towards this, exosomes were isolated from CSF from 13 patients with anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis, 11 patients with anti-gamma-aminobutyric acid-B (GABAB) receptor encephalitis, 9 patients with anti-leucine-rich glioma-inactivated 1 (LGI1) encephalitis, and 8 patients with anti-contactin-associated protein-like 2 (CASPR2) encephalitis, and 12 control individuals negative of antibodies against neuronal autoantigens. There were ten miRNAs highly expressed in patients with anti-NMDAR encephalitis compared to those in control subjects. Eight miRNAs were found to be lower expressed in anti-NMDAR encephalitis CSF-derived exosomes. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways enriched by AE differential expressed exosomic miRNAs demonstrated that AE-related exosomic miRNAs may participate as a feedback regulation in cancer development. In addition, the exosome concentration in CSF of 9 HSE patients was significantly higher compared to those from 9 HSV( -) patients. This observation was consistent with the results that exosome concentration was found to be higher in the animal model which was inoculated intranasally with HSV-1 compared to controls. Furthermore, western blot demonstrated that the subunits of NMDAR, GABABR, and AMPAR were detected highly expressed in exosomes derived from sera of HSV-1-treated animal model compared to controls. More importantly, exosomes isolated from CSF of HSE patients contained higher expression levels of two miRNAs encoded by HSV, miR-H2-3p, and miR-H4-3p compared to those from HSV( -) patients. In summary, HSV may trigger brain autoimmunity in HSE by presentation of surface autoantigens via exosomes.

The Efficacy of High- or Low-Frequency Transcranial Magnetic Stimulation in Alzheimer's Disease Patients with Behavioral and Psychological Symptoms of Dementia.

INTRODUCTION: Alzheimer's disease (AD) is usually accompanied by different degrees of behavioral and psychological symptoms of dementia (BPSD). Transcranial magnetic stimulation (TMS) has been applied for the treatment of AD as a painless and noninvasive therapy. However, the efficacy of repetitive TMS (rTMS) with different frequencies in AD with BPSD remains unknown. METHODS: A total of 32 AD patients with psychobehavioral symptoms were selected as the study subjects. Among them, 16 patients were included in the high-frequency TMS group with an average disease duration of 6.22 ± 2.55 years. The low-frequency TMS group was gender and age matched with a disease course of 7.02 ± 3.33-year average duration. The high-frequency TMS group received TMS treatment twice per day for 4 weeks under 80% MT stimulation intensity, 10-Hz frequency for 0.5 h each time, and the low-frequency TMS group received TMS treatment of 2-Hz frequency for 0.5 h each time. Neuropsychological status was assessed by the Behavioral Pathology in Alzheimer's Disease Rating Scale (BEHAVE-AD) score. The behavioral ability was assessed by the Abilities of Daily Living (ADL) scale; cognitive function was evaluated by Mini-Mental State Examination (MMSE). The levels of ß amyloid 40 and 42 (Aß40 and Aß42) in plasma were detected using a double-antibody sandwich enzyme-linked immunosorbent assay. All patients underwent brain magnetic resonance imaging (MRI) before and after the experiment. RESULTS: After 2 weeks of treatment, the BEHAVE-AD and ADL scores of the patients in the high-frequency group were significantly lower than those before the treatment, and they continued to decrease after 4 weeks of treatment. The BEHAVE-AD and ADL scores of the low-frequency TMS group were also significantly lower than before treatment. The comparison between groups at different time points showed that the BEHAVE-AD and ADL scores of the patients in the high-frequency group were significantly lower than those of the patients in the low-frequency TMS group. The MMSE of high-frequency TMS-treated patients increased from 14.22 ± 3.55 before treatment to 14.67 ± 2.22 at 2 week's treatment and 17.33 ± 3.11 at 4 week's treatment (p < 0.01) in contrast to 14.19 ± 3.47, 14.28 ± 3.41, and 14.49 ± 2.79, respectively, found in the low-frequency TMS group. At week 4, the high-frequency TMS-treated group's plasma Aß40 did not change compared to that before treatment. No effects on plasma Aß42 were observed between the high- vs. low-frequency TMS groups. The incidence of adverse reactions during treatment was comparable between groups. CONCLUSION: These results indicate that high-frequency TMS has the advantages of fast results, good efficacy, and high safety for the treatment of psychobehavioral abnormalities in AD patients. In addition, our study suggests that high-frequency TMS intervention can further improve the cognitive function of AD patients.

Exosomes isolated during dopaminergic neuron differentiation suppressed neuronal inflammation in a rodent model of Parkinson's disease.

Our previous investigation showed Wnt signal pathway was significantly activated during DA neuron differentiation of epiblast-derived stem cells. In this study, we next attempt to examine the therapeutic potential of the purified exosomes derived bone marrow mesenchymal stem cells (BMSCs) by administrating exosomes into the rat striatum of parkinson's disease (PD) animal model. Results revealed that the protein levels of interleukin (IL)-6, IL-1ß, tumor necrosis factor-alpha (TNF-α), and reactive oxygen species (ROS) in the substantia nigra of PD rats were down regulated after injection of BMSC induced-Exosomes into the striatum of PD model compared to BMSC quiescent-Exosomes. In addition, the expression of ionized calcium binding adaptor molecule 1 (Iba1) mRNA was significantly decreased, while the expression of tyrosine hydroxylase (TH) mRNA was increased after injection of BMSC induced-Exosomes. Injection of BMSC induced-Exosomes into the striatum rescued the rotation behavior and climbing speed in the PD rats. More importantly, Wnt5a was found to be enriched in BMSC induced Exosomes, which could be effectively transferred to the substantia nigra of PD rats. In conclusion, these findings demonstrated that exosomes isolated during dopaminergic neuron differentiation could rescue the pathogenic features of Parkinson's disease by reshaping the inflammatory microenvironment in the substantia nigra and repairing the injury to DA nerves.

Neuroprotective Effect of Trans-Resveratrol in Mild to Moderate Alzheimer Disease: A Randomized, Double-Blind Trial.

INTRODUCTION: Amyloid-beta (Aß) protein is a major component of the extracellular plaque found in the brains of individuals with Alzheimer's disease (AD). In this study, we investigated the effect of trans-resveratrol as an antagonist treatment for moderate to mild AD, as well as its safety and tolerability. METHODS: This was a case-control study that enrolled 30 selected patients who had been clinically diagnosed with moderate to mild AD. These patients were randomly divided into two groups, namely, a placebo group (n = 15) and a trans-resveratrol group (n = 15) who received 500 mg trans-resveratrol orally once daily for 52 weeks. Brain magnetic resonance imaging (MRI) examinations were performed on and cerebrospinal fluid (CSF) samples were obtained from all participants before (baseline) and after the study (52 weeks). Enzyme-linked immunosorbent assays were used to determine the levels of plasma Aß40 and Aß42 and CSF Aß40 and Aß42. RESULTS: The results showed that the changes over the study period in the levels of Aß40 in the blood and CSF of the patients treated with trans-resveratrol were not statistically significant (P > 0.05). In contrast, patients who received placebo showed a significant decrease in Aß40 levels compared with that at the beginning of the study (CSF Aß40: P = 0.024, plasma Aß40: P = 0.036). Analysis of the images on the brain MRI scans revealed that the brain volume of the patients treated with trans-resveratrol was significantly reduced at 52 weeks (P = 0.011) compared with that of patients in the placebo treatment group, Further analysis indicated that the level of matrix metallopeptidase 9 in the CSF of the patients treated with trans-resveratrol at 52 weeks decreased by 46% compared with that of patients in the placebo group (P = 0.033). CONCLUSION: These results indicate that trans-resveratrol has potential neuroprotective roles in the treatment of moderate to mild AD and that its mechanism may involve a reduction in the accumulation and toxicity of Aß in the brain of patients, thereby reducing neuroinflammation. TRIAL REGISTRATION: Chinese clinical trial registry: CTR20151780X.

Normal-sized basal ganglia perivascular space related to motor phenotype in Parkinson freezers.

Changes in basal ganglia (BG) perivascular spaces (PVSs) are related to motor and cognitive behaviors in Parkinson's disease (PD). However, the correlation between the initial motor phenotype and PVSs distribution/burden in PD freezing of gait (FOG) remains unclear. In addition, the normal-sized PVSs (nPVSs) have not been well-studied. With high-resolution 7T-MRI, we studied nPVSs burden in BG, thalamus, midbrain and centrum semiovale. The numbers and volume of nPVSs were assessed in 10 healthy controls, 10 PD patients without FOG, 20 with FOG [10 tremor dominant (TD), 10 non-TD subtype]. Correlation analyses were further performed in relation to clinical parameters. In this proof of concept study, we found that the nPVS burden of bilateral and right BG were significantly higher in freezers. A negative correlation existed between the tremor score and BG-nPVSs count. A positive correlation existed between the levodopa equivalent daily dose and BG-nPVSs count. The nPVS burden correlated with the progression to FOG in PD, but the distribution and burden of nPVS differ in TD vs. non-TD subtypes. High resolution 7T-MRI is a sensitive and reliable tool to evaluate BG-nPVS, and may be a useful imaging marker for predicting gait impairment that may evolve into FOG in PD.

Altered microRNAs in cerebrospinal fluid exosomes in paraneoplastic and autoimmune encephalitis: A possible feedback in cancer development.

AIMS: The purpose of this study was to examine the role of cerebrospinal fluid (CSF) exosomes from patients with paraneoplastic and autoimmune encephalitis (AE). MAIN METHODS: Towards this, microRNA profiling in the exosomes which were isolated from cerebrospinal fluid of 12 patients with anti-N-methyl-d-aspartate (NMDA) receptor encephalitis, 12 patients with anti-gamma-aminobutyric acid-B (GABAB) receptor encephalitis, 12 patients with anti-leucine-rich glioma-inactivated 1 (LGI1) encephalitis, and 12 patients with anti-contactin-associated protein-like 2 (CASPR2) encephalitis, and 12 control individuals negative of antibodies against neuronal auto-antigens. Selected findings were validated with quantitative RT-PCR. DIANA-mirPath was chosen for bioinformatic analysis. KEY FINDINGS: There were ten miRNAs higher expressed in AE patients with anti-NMDAR encephalitis compared to those in healthy controls. Further, eight miRNAs were found to be lower expressed in anti-NMDAR encephalitis CSF derived exosomes. In addition, Endometrial cancer, p53 signaling pathway, Non-small cell lung cancer, Small cell lung cancer, Transcriptional misregulation in cancer, Basal cell carcinoma, Acute myeloid leukemia, Renal cell carcinoma, Colorectal cancer, Choline metabolism in cancer, Melanoma, Pancreatic cancer, Prostate cancer, Ras signaling pathway, Glioma, Pathways in cancer, and Proteoglycans in cancer (all p < 0.01) were significantly enriched in differentially expressed miRNAs. SIGNIFICANCES: Exosomes expressing specific miRNAs in antibody positive AE may participate as a feedback regulation in cancer development.

Recent Advances on Extracellular Vesicles in Central Nervous System Diseases.

Extracellular vesicles (EVs) are particles released by multiple cells, encapsulated by lipid bilayers and containing a variety of biological materials, including proteins, nucleic acids, lipids and metabolites. With the advancement of separation and characterization methods, EV subtypes and their complex and diverse functions have been recognized. In the central nervous system (CNS), EVs are involved in various physiological and pathological processes, such as regulation of neuronal firing, synaptic plasticity, formation and maintenance of myelin sheath, propagation of neuroinflammation, neuroprotection, and spread and removal of toxic protein aggregates. Activity-dependent alteration of constituents enables EVs to reflect the change of cell and tissue states, and the wide distribution of EVs in biological fluids endows them with potential as diagnostic and prognostic biomarkers for CNS diseases, including neurodegenerative disease, cerebrovascular disease, traumatic brain disease, and brain tumor. Favorable biocompatibility, ability of crossing the blood-brain barrier and protecting contents from degradation, give promising therapeutic effects of EVs, either collected from mesenchymal stem cells culture conditioned media, or designed as drug delivery vehicles loaded with specific agents. In this review, we summarized EVs' basic biological properties, and mainly focused on their applications in CNS diseases.

Exosomes expressing neuronal autoantigens induced immune response in antibody-positive autoimmune encephalitis.

The immunological role of exosomes in autoimmune encephalitis (AE) remains uncharacterized and not examined. In this study we ought to determine whether exosomes are generated in AE and to define the presence of cell surface neuronal autoantigens (autoAgs) in the cargo. Exosomes were isolated from cerebrospinal fluid (CSF) from 12 patients with anti-N-methyl-d-aspartate (NMDA) receptor encephalitis, 8 patients with anti-gamma-aminobutyric acid-B (GABAB) receptor encephalitis, 8 patients with anti-leucine-rich glioma-inactivated 1 (LGI1) encephalitis, 8 patients with anti-contactin-associated protein-like 2 (CASPR2) encephalitis, 10 patients with anti-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid 1,2 (AMPA) receptor encephalitis and 30 control individuals negative of antibodies against neuronal autoAgs. Western blot demonstrated that CSF or sera derived exosomes from AE contained specific neuronal autoAgs in protein aggregates, however, control subjects had no detectable levels of these neuronal autoAgs. In addition, development of antibodies against NMDAR, GABABR, LGI1, CASPR2, and AMPAR were detected in the sera after 30 days immunization of C57BL/6 J mice with exosomes isolated from antibody positive AE patients; Enzyme-linked immunospot (ELISpot) assay demonstrated increased frequency of neuronal autoAgs-specific IL-17 and IFN-γ in splenocytes from AE derived exosomes immunized mice. We concluded that exosomes expressing neuronal autoAgs were present in CSF from antibody positive AE patients, and we propose these exosomes carrying neuronal autoAgs would play an important role in the immune pathogenesis of autoimmune encephalitis.

Differential Expression of microRNA Profiles and Wnt Signals in Stem Cell-Derived Exosomes During Dopaminergic Neuron Differentiation.

The role of secreted exosomes during dopaminergic (DA) neuron differentiation is still unknown. To investigate the roles of exosomes in DA neuron fate specification, we profiled exosomal microRNAs (miRNAs) during DA neuron differentiation of epiblast-derived stem cells (EpiSCs). There were 26 miRNAs differentially expressed (relative fold >2, p < 0.05) in EpiSC-derived exosomes at 0, 2, 4, 6, 8, 10, 12, and 14 days of DA epiblast differentiation. Among them, 23 exosomic miRNAs were significantly increased, including miR-124, miR-132, miR-133b, miR-218, miR-9, miR-34b, miR-34c, and miR-135a2, while three exosomic miRNAs (miR-214, miR-7a, and miR-302b) were decreased, when compared with control samples. Bioinformatics analysis by DIANA-mirPath demonstrated that extracellular matrix-receptor interaction, signaling pathways regulating pluripotency of stem cells, FoxO signaling pathway, DA synapse, Wnt signaling pathway, GABAergic synapse, and neurotrophin signaling pathway were significantly enriched in DA differentiation-related miRNA signature (all p-values <0.012). Furthermore, messenger RNAs for nine DA neuronal markers tyrosine hydroxylase (TH), Nr4a2, Pitx3, Drd1a, Lmx1a, Lmx1b, Foxa1, Dmrt5, and Slc18a2 were significantly increased expressed over time in exosomes derived from differentiated EpiSCs. Interestingly, adding with exosomes derived from EpiSC induction experiment resulted in a twofold increase of TH-positive neurons production (35% vs. 17%, p < 0.01) during DA neuronal differentiation from mouse embryonic stem cells (ESCs). In summary, our results suggested exosomal miRNAs are potential regulators of DA neuron differentiation. More importantly, EpiSC-derived exosomes could promote the generation of DA neuron differentiation from ESCs.

Characterization of the 18 kDa translocator protein (TSPO) expression in post-mortem normal and Alzheimer's disease brains.

The 18 kDa translocator protein (TSPO) is a widely used target for microglial PET imaging radioligands, but its expression in post-mortem normal and diseased human brain is not well described. We aimed at characterizing the TSPO expression in human control (CTRL) and Alzheimer's disease (AD) brains. Specifically, we sought to: (1) define the cell type(s) expressing TSPO; (2) compare tspo mRNA and TSPO levels between AD and CTRL brains; (3) correlate TSPO levels with quantitative neuropathological measures of reactive glia and AD neuropathological changes; and (4) investigate the effects of the TSPO rs6971 SNP on tspo mRNA and TSPO levels, glial responses and AD neuropathological changes. We performed quantitative immunohistochemistry and Western blot in post-mortem brain samples from CTRL and AD subjects, as well as analysis of publicly available mouse and human brain RNA-Seq datasets. We found that: (1) TSPO is expressed not just in microglia, but also in astrocytes, endothelial cells and vascular smooth muscle cells; (2) there is substantial overlap of tspo mRNA and TSPO levels between AD and CTRL subjects and in TSPO levels between temporal neocortex and white matter in both groups; (3) TSPO cortical burden does not correlate with the burden of activated microglia or reactive astrocytes, Aß plaques or neurofibrillary tangles, or the cortical thickness; (4) the TSPO rs6971 SNP does not significantly impact tspo mRNA or TSPO levels, the magnitude of glial responses, the cortical thickness, or the burden of AD neuropathological changes. These results could inform ongoing efforts toward the development of reactive glia-specific PET radioligands.

Application of Neck Circumference in Four-Variable Screening Tool for Early Prediction of Obstructive Sleep Apnea in Acute Ischemic Stroke Patients.

BACKGROUND: The purpose of this study was to validate and pilot the use of the four-variable screening tool (4V) and modified 4V tools to identify acute ischemic stroke and transient ischemic attack (TIA) patients at high risk of obstructive sleep apnea (OSA). METHODS: Two modified scales, 4V-1 (ie, using neck circumference instead body mass index, regardless of gender) and 4V-2 (ie, as above but scored differently according to gender) were designed. These tools were used in a consecutive cohort of 124 acute ischemic stroke/TIA patients, together with the 4V-1, 4V-2, 4V, as well as the STOP-BANG, the Berlin questionnaire, and the Epworth Sleepiness Scale (ESS). Objective level 2 or level 3 polysomnography was used to confirm OSA and its severity. Both questionnaires and polysomnography were completed within 1 week from symptom onset. RESULTS: Area under the curve (AUC) of 4V was 0.807 (P< .0001) while AUC of STOP-BANG, Berlin Questionnaire and ESS were .701 (P< .0001), .704 (P< .0001) and .576 (P = .1556), respectively. AUC of 4V was greater than of STOP-BANG (z = 2.200, P = .0220), Berlin (z = 2.024, P = .0430) and ESS (z = 3.363, P = .0003). AUC of modified 4V-1 and modified 4V-2 were .824 (P< .001) and .835 (P< .001), respectively. Performance of modified 4V-2 was higher versus modified 4V-1 (z = 2.111, P = .0348) and higher but not significantly so to regular 4V (z = 1.784, P = .0744). CONCLUSIONS: Neck circumference scored by gender is a useful substitution to body mass index in the 4V when screening OSA at early stages of ischemic stroke/TIA patients.

Using Extracellular Circulating microRNAs to Classify the Etiological Subtypes of Ischemic Stroke.

There is no effective biological method to classify ischemic stroke subtypes. In this study, we first performed a systematical gene array study on serum microRNAs with different ischemic stroke subtypes including 13 normal control subjects (NCs) and 87 ischemic stroke (IS) patients including 23 cardioembolism (CARD), 26 large artery atherosclerosis (LAA), 27 lacunar infarct (LAC), and 11 stroke of undetermined etiology (SUE). Validation was performed by using an independent cohort of 20 NCs and 85 IS patients including 28 CARD, 23 LAA, 18 LAC, and 16 SUE. In the pilot discovery gene array study, we found specific serum microRNA signatures between different ischemic stroke subtypes (CARD, LAA, LAC, and SUE). We further validated 6 microRNAs [miR-125b, miR-125a, let-7b, let-7e, miR-7-2-3p, miR-1908] in a different group of ischemic stroke subtypes by using an independent cohort of 20 NCs, 28 CARD, 23 LAA, 18 LAC, and 16 SUE. Moreover, these circulating miRNAs were further detected to be differentially expressed between pre- vs. post-stroke in different ischemic stroke subtypes. The ROC analysis showed that miR-125b, miR-125a, let-7b, and let-7e could discriminate CARD patients from normal controls and other subtypes. Furthermore, ROC curves shown that miR-7-2-3p and miR-1908 showed significant area-under-the-curve values in both LAA and LAC patients. In conclusion, these results demonstrated that circulating miRNAs in sera could be potentially novel risk factors that involve in the pathogenesis of ischemic stroke subtypes.

NFE2L2 variations reduce antioxidant response in patients with Parkinson disease.

Oxidative stress has been recognized as a risk factor of Parkinson's disease (PD) development. We hypothesized that decreased function of the nuclear factor (erythroid-derived 2)-like 2 (NFE2L2)-antioxidant response element (ARE) pathway might predispose to Parkinsonism. A case-control study was performed between NFE2L2 Single Nucleotide Polymorphism (SNP) and PD in a cohort of 765 unrelated patients with diagnosis of PD and 489 matched normal individuals. We found that c.351T>A, D117E (P = 0.003, OR = 2.8) and c.351T>A, D117E (P = 0.012, OR = 1.9) were significantly associated with PD. The risk allele of both polymorphisms showed a high frequency in our PD sample (c.351A: 19.7% and c.423T: 7.8%). The association between both c.351T>A and c.423G>T and PD was further confirmed in an independent case-control cohort consisting of 210 individuals with PD and 148 normal controls. We further found that over expression of D117E and Q141H variants of NFE2L2 reduced target genes expression of Glutathione S-transferase Pi 1 (GSTP1), Glutathione S-transferase Mu 1 (GSTM1), and Heme oxygenase 1 (HO-1) genes. NFE2L2 D117E and Q141H impaired activation of ARE-driven transcriptional activity. Our findings indicate that NFE2L2 may play an important role in the pathogenesis of PD in Chinese populations.

Altered microRNA profiles in cerebrospinal fluid exosome in Parkinson disease and Alzheimer disease.

The differential diagnosis of Parkinson's diseases (PD) is challenging, especially in the early stages of the disease. We developed a microRNA profiling strategy for exosomal miRNAs isolated from cerebrospinal fluid (CSF) in PD and AD. Sixteen exosomal miRNAs were up regulated and 11 miRNAs were under regulated significantly in PD CSF when compared with those in healthy controls (relative fold > 2, p < 0.05). MiR-1 and miR-19b-3p were validated and significantly reduced in independent samples. While miR-153, miR-409-3p, miR-10a-5p, and let-7g-3p were significantly over expressed in PD CSF exosome. Bioinformatic analysis by DIANA-mirPath demonstrated that Neurotrophin signaling, mTOR signaling, Ubiquitin mediated proteolysis, Dopaminergic synapse, and Glutamatergic synapse were the most prominent pathways enriched in quantiles with PD miRNA patterns. Messenger RNA (mRNA) transcripts [amyloid precursor protein (APP), α-synuclein (α-syn), Tau, neurofilament light gene (NF-L), DJ-1/PARK7, Fractalkine and Neurosin] and long non-coding RNAs (RP11-462G22.1 and PCA3) were differentially expressed in CSF exosomes in PD and AD patients. These data demonstrated that CSF exosomal RNA molecules are reliable biomarkers with fair robustness in regard to specificity and sensitivity in differentiating PD from healthy and diseased (AD) controls.

Evidence for polymerase gamma, POLG1 variation in reduced mitochondrial DNA copy number in Parkinson's disease.

BACKGROUND: It remains unclear whether the mtDNA content is related to the clinicopathological prognosis in Parkinson's disease (PD). METHODS/RESULTS: We analyzed the copy number of mtDNA using quantitative real-time PCR in 414 cases with PD and 231 healthy subjects from mainland of China. The level of mtDNA was significantly decreased in PD patients' peripheral blood as compared to that of healthy controls (p < 0.001). Furthermore, lower mtDNA copy number was more frequently detected (75%) in the older onset age group (≥ 50 years old) than that in (49%) the younger group (<50 years old, p = 0.007), suggesting mtDNA content might be an important genetic event in PD progression. Using direct sequencing, we examined the mutations in the D-loop region of mtDNA in 318 PD patients. The results revealed that 17% of the PD patients carried mutations in the D-loop of mtDNA, and 55% of these mutations were heteroplasmic. In addition, PD patients harboring AA + AA genotype of c.2070-12T > A and c.2070-64G > A in POLG1 along with mutations in POLG1 had a significantly lower copy number of mtDNA than those of PD patients without POLG1 alterations. CONCLUSIONS: Our results provided evidence for a significantly lower of mtDNA copy number in PD patients and POLG1 variation for reducing mtDNA copy number in PD.