Metabolites of intestinal fora can be used as diagnostic and progressive markers for mild cognitive impairment.

Purpose: The aim of the work was to analyze the metabolites of the intestinal microbiota from the patients with mild cognitive impairment (MCI) and progressive MCI due to Alzheimer's disease (AD). Method: Two cohorts were established. The first one included 87 subjects with 30 healthy controls (NC), 22 patients with MCI due to AD, and 35 patients with AD. The second cohort included 87 patients with MCI due to AD, who were followed up for 2 years and finally were divided into progressive MCI due to AD group (P-G) and unprogressive MCI due to AD group (U-G) according their cognitive levels. Fecal samples were collected to all patients at the baseline time point. Differential metabolites were subjected to pathway analysis by MetaboAnalyst. Results: In the first cohort, we found 21 different metabolites among the three groups (AD, MCI, and NC). In the second cohort, we identified 19 differential metabolites between the P-G and U-G groups. By machine learning analysis, we found that seven characteristic metabolites [Erythrodiol, alpha-Curcumene, Synephrine, o-Hydroxylaminobenzoate, 3-Amino-4-hydroxybenzoic acid, 2-Deoxystreptamine, and 9(S] were of characteristic significance for the diagnosis of MCI due to AD, and six metabolites (Indolelactate, Indole-3-acetaldehyde, L-Proline, Perillyl, Mesaconate, and Sphingosine) were the characteristic metabolites of early warning for the progression of MCI due to AD. D-Glucuronic acid was negatively correlated with Apolipoprotein E4 (APOE4). Perillyl alcohol was negatively correlated with all of the five biomarkers [P-tau181, Neurofilament light chain (NF-light), Aß1-42, Aß1-40, and glial fibrillary acidic protein (GFAP)], but Indoleacetaldehyde was positively correlated with three biomarkers (P-tau181, Aß1-42, and GFAP). Three characteristic metabolites (3-Amino-4-hydroxybenzoate, 2-Deoxystreptamine, and p-Synephrine) were positively correlated with Aß1-42. 2-Deoxystreptamine, 9(S)-HPOT, and Indoleacetaldehyde were positively correlated with GFAP. L-Proline and Indoleacetaldehyde were positively correlated with NF-light. Conclusion: Specific metabolites of intestinal fora can be used as diagnostic and progressive markers for MCI.

Relevance of cerebral small vessel disease load scores in first-ever lacunar infarction.

AIM: To reveal the correlation between total cerebrovascular disease load and primary lacunar infarction. BACKGROUND: Cerebral small vessel disease (CSVD) is the lack of specific clinical manifestations, whose clinical diagnoses are highly dependent on neuroimaging results. Total CSVD load scores may be more suitable for the assessment of overall brain function damage caused by CSVD. Little is known about whether the association between imaging markers of CSVD and CSVD total load scores at the time of first-ever lacunar infarction (LI). METHODS: clinical data of 396 patients hospitalised from September 2016 to May 2018 due to a first-ever LI (case group), along with patients diagnosed with CSVD based on imaging alone and those with no abnormalities (control group) based on magnetic resonance imaging (MRI). Binary logistic regression and multiple ordered logistic regression were used to analyse the characteristics of imaging markers of CSVD in patients with first-ever LI, including different total score burden and distribution, and the relationship between different markers. RESULTS: In 396 patients, smoking, cholesterol level and total small vessel disease (SVD) score were all significantly associated with the first-ever LI. There were more LI, cerebral microbleeds (CMB), white matter hyperintensities (WMH), and moderate to severe enlarged perivascular spaces (EPVS) in the first-ever LI group, relative to controls (p < 0.01). The Fazekas scores for periventricular WMH, deep WMH, and total Fazekas score were all significantly higher in patients with first-ever LI relative to those with no cerebral abnormalities (p < 0.01). An analysis of various imaging markers of CSVD revealed a significant correlation between the presence and degree of any marker and the severity of other markers, even after adjusting for the presence of other markers (p < 0.05). CONCLUSIONS: The first-ever LI group exhibited higher total CSVD score loads, a greater number of lacunae, CMB, severe WMH and moderate to severe EPVS. Smoking is an independent risk factor in patients with first-ever LI.

Spinal cord injury and its underlying mechanism in rats with temporal lobe epilepsy.

Numerous cases of spinal cord injury following seizure have been previously reported. However, whether spinal cord injury is a common occurrence after seizures and its underlying mechanisms remain unclear. The present study generated a Sprague-Dawley rat model of temporal lobe epilepsy (TLE), and Nissl staining and transmission electron microscopy were used to detect tissue damage. In addition, Evans blue staining was used to detect damage to the blood-brain barrier (BBB) and albumin extravasation. In addition, double-staining was used to detect the association between neurons and extravasated albumin. Furthermore, neuronal degeneration was assessed using Fluoro-Jade C staining, while fluorescence staining and western blotting were used to detect apoptosis and inflammation. In the present study, spinal cord injury was only observed in rats with grade IV-V seizures, whereas Nissl staining showed structural damage and decreased neuronal cell numbers in the brain and the spinal cord. The present study identified BBB damage and albumin extravasation in rats of the TLE groups. Double-staining for albumin and neurons showed a significant match of neurons positive for albumin. Fluoro-Jade C staining indicated neuronal degeneration in the brain, but not the spinal cord in the TLE rats. High levels of caspase-3 were also detected in the injured spinal cord. A small number of albumin+ neurons in the spinal cord presented caspase-3+ signals in rats of the TLE groups. The expression levels of intercellular adhesion molecule 1, CD11b and inflammatory factors such as tumor necrosis factor-α and interleukin-6 were significantly elevated in the injured spinal cord. The present results suggested that spinal cord injury occurred in rats as a result of severe seizure attacks, and that BBB damage, albumin extravasation, inflammation and apoptosis contributed to the pathological changes observed during spinal cord injury.

Effects of the transcription factor Olig1 on the differentiation and remyelination of oligodendrocyte precursor cells after focal cerebral ischemia in rats.

The differentiation and maturation of oligodendrocyte precursor cells (OPCs) is important for remyelination in the central nervous system. Nevertheless, this process is often limited and incomplete in ischemic injury. Oligodendrocyte transcription factor 1 (Olig1) is important for the maturation of OPCs and the repair of demyelinated lesions. However, how Olig1 modulates the development of OPCs or the remyelination associated with ischemic injury remains unclear. The present study aimed to examine alterations in OPCs, and the expression of myelin and Olig1, at different time-points after focal cerebral ischemia using immunohistochemistry and western blot techniques to elucidate the role of Olig1 in the maturation of OPCs and remyelination. The present results showed that the expression of Olig1 significantly decreased at 1 day after middle cerebral artery occlusion (MCAO) and returned to normal levels from day 3 to 28. Additionally, Olig1 was found to translocate into the nucleus following ischemia in the brain. The number of OPCs in the ischemic striatum significantly declined at days 1 and 3 following MCAO, and increased at days 7, 14 and 28 compared with the control. The expression of myelin basic protein, a marker of mature oligodendrocytes and myelin, gradually decreased from day 1 to 7 after ischemia and recovered at day 14 and 28; however, the levels were lower than those in the control group. The present results indicated that the restored normal level of Olig1 following ischemia may play an important role in the maturation of OPCs through its translocation into the nucleus, where it may promote the growth and development of myelin under pathological conditions. However, this endogenous recovery mechanism fails to fully repair the demyelinated lesion. The data of the present study may help clinicians understand the expression pattern of Olig1 and its potential role in endogenous remyelination after ischemia, which may have implications for the treatment of diseases that lead to demyelination.

What influenced the lesion patterns and hemodynamic characteristics in patients with internal carotid artery stenosis? A retrospective study.

OBJECTIVES: This study aimed to explore the dynamic changes of lesion patterns and hemodynamic characteristics in patients with internal carotid artery stenosis (ICAS). PATIENTS AND METHODS: Patients who had suffered an acute ischemic stroke in the distribution of ipsilateral ICAS were included. Computed tomography (CT) and transcranial doppler ultrasound (TCD) were conducted to evaluate the degree of ICAS and the hemodynamic characteristics of the intracranial and extracranial arteries. RESULT: A total of 424 patients were included in the study. With the aggravation of ICAS, blood velocity in ipsilateral ICA was increased, while blood flow in the ipsilateral middle cerebral artery (MCA) was decreased. In the same degree of ICAS, patients with opened communicating arteries showed relatively higher blood perfusion in MCA compared with those without communicating arteries. In the average stage of ICAS, small lesions (D=0-1.5cm), middle lesions (1.5cm3.0cm) commonly existed. The number of small and large lesions significantly increased when the blood flow of ipsilateral MCA decreased. In the same degree of stenosis, the number of small lesions and large lesions, and the total area of all lesions, evidently increased with the decrease of ipsilateral MCA blood velocity. CONCLUSION: Hypoperfusion is an independent risk factor for ischemic lesions in patients with ICAS. Whether or not the communicating arteries are open influences the blood flow of the intracranial arteries. TCD was a convenient and rapid tool to assess intracranial perfusion and vascular compensatory status.

Relationship between CHOP/GADD153 and unstable human carotid atherosclerotic plaque.

BACKGROUND AND AIMS: The signaling protein C/EBP homologous protein (CHOP) and corresponding growth-arrest-and-DNA-damage-inducible gene 153 (GADD153) is associated with endoplasmic reticulum stress (ERS), which can lead to apoptosis. Our study aims to elucidate the role of CHOP/GADD153 in unstable atherosclerotic (AS) plaque formation isolated from confounding factors such as diabetes mellitus, primary hyperlipidemia, autoimmune deficiencies/abnormalities, essential hypertension, chronic heart failure, chronic kidney disease, and smoking. MATERIAL AND METHODS: We collected carotid artery tissue samples from patients aged 50-80 years-old who received carotid endarterectomies (CEA) at our institution. We obtained fresh AS plaque samples during CEA and preserved the specimens immediately in the operating room with liquid nitrogen. Samples were categorized as stable or unstable AS plaques according to a six-stage histologic classification. CHOP/GADD153 expression was then examined with immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). RESULTS: A total of 32 patients met our inclusion and exclusion criteria, with 24 (75.0%) classified as unstable lesions. The mean optical density ratio normalized to GAPDH for CHOP/GADD153 in stable and unstable groups was 0.357 ± 0.025 and 0.490 ± 0.027, respectively (p < .05). Positive immunostaining of CHOP/GADD153 was found in macrophages and smooth muscle cells of unstable AS plaques with a mean integrated optical density of 0.63 ± 0.03, compared to 0.17 ± 0.05 in the stable group (p < .05). CONCLUSIONS: In conclusion, we were able to show significant elevation of CHOP/GADD153 in unstable plaques independent of other confounding factors that induce ERS.

Neuronal uptake of serum albumin is associated with neuron damage during the development of epilepsy.

It is well established that brain blood barrier dysfunction following the onset of seizures may lead to serum albumin extravasation into the brain. However, the effect of albumin extravasation on the development of epilepsy is yet to be fully elucidated. Previous studies have predominantly focused on the effect of albumin absorption by astrocytes; however, the present study investigated the effects of neuronal uptake of albumin in vitro and in kainic acid-induced Sprague-Dawley rat models of temporal lobe epilepsy. In the present study, electroencephalogram recordings were conducted to record seizure onset, Nissl and Evans blue staining were used to detect neuronal damage and albumin extravasation, respectively, and double immunofluorescence was used to explore neuronal absorption of albumin. Cell counting was also conducted in vitro to determine whether albumin contributes to neuronal death. The results of the present study indicated that extravasated serum albumin was absorbed by neurons, and the neurons that had absorbed albumin died and were dissolved 28 days after seizure onset in vivo. Furthermore, significant neuronal death was detected after albumin absorption in vitro in a dose- and time-dependent manner. These results suggested that albumin may be absorbed by neurons following the onset of seizures. Furthermore, the results indicated that neuronal albumin uptake may be associated with neuronal damage and death in epileptic seizures. Therefore, attenuating albumin extravasation following epileptic seizures may reduce brain damage and slow the development of epilepsy.

Leukocyte Infiltration Triggers Seizure Recurrence in a Rat Model of Temporal Lobe Epilepsy.

Epilepsy, which affects about 1 % of the population worldwide, leads to poor prognosis and increased morbidity. However, effective drugs providing satisfactory control on seizure relapse were rare, which encouraged more etiological studies. Whether inflammation is one of key events underlying seizure is in debate. In order to explore the role of inflammatory in the pathogenesis and development of epilepsy, we conducted intra-caudal vein injection of leukocytes to aggravated brain inflammatory process in kainic acid-induced seizure model in this study. The results showed that intravenous administration of activated leukocytes increased the frequency and reduced the latent phase of seizure recurrences in rat models of epileptic seizure, during which leukocyte inflammation, brain-blood barrier damage, and neuron injury were also significantly aggravated, indicating that leukocyte infiltration might facilitate seizure recurrence through aggravating brain inflammation, brain-blood barrier damage, and neuron injury.

Inactivition of CDKL3 mildly inhibits proliferation of cells at VZ/SVZ in brain.

CDKL3 has an important role in regulating cell growth and/or differentiation, and its inactivation is recently reported to be related to non-syndromic mild mental retardation (MR). MR is a common neurological disorder, predominantly characterized by impaired cognitive function. Though genetic factors play a very important role in the pathogenesis of MR, to date, only few genes linked to MR have been characterized and understood very well. Here, we investigated the role of the CDKL3 in the proliferation of cells surrounding the brain ventricle, and the results showed down-regulating CDKL3 by the method of RNAi in the cells surrounding the brain ventricle of the mouse embryo at E15 may inhibit their proliferation. As our previous study had shown that Cdkl3 mRNA expression is developmentally regulated in the central nervous system, peaking during late embryonic and early postnatal stages which are the key stages of neurite formation and maturation, furtherly, the present findings indicated that CDKL3 may be involved in proliferation of cells surrounding the brain ventricle where neuronal progenitor cells are enriched during the late embryo stage, supporting the notion that CDKL3 inactivation contributes to non-syndromic mild MR.

The activity of the Ang/Tie-2 system in the brain that suffered acute carbon monoxide poisoning.

Acute carbon monoxide poisoning (ACMP) leads to significant toxicity of the central nervous system and heart, and even death, following it, some patients suffered delayed encephalopathy. Until now, no theory had explained it exactly. It was reported that neovascularization was found in acute ischemic brains and also that angiopoietins (Ang) play important roles in the process of angiogenesis, for example, the members of Ang family, Ang-1 and Ang-2 may promote angiogenesis by combining with endothelial-specific cell surface tyrosine kinase receptor Tie-2. Interestingly, some studies suggested that small vascular injury may play an important role in the pathogenesis of delayed encephalopathy after carbon monoxide poisoning. Does neovascularization also occur in the brains after ACMP? Do Ang also take part in the pathologic processes in the brains that suffered ACMP? People know little about it. In the present study, we showed that neovascularization also occurred in the brains that suffered ACMP, and there are two expression peaks of Ang-1, Ang-2 and Tie-2, respectively, in the mice brains on the 3rd day and the 7th day following ACMP, and draw a conclusion that the Ang/Tie-2 system takes part in the pathologic processes in the brains that suffered ACMP by participating in neovascularization.

Non-syndromic mild mental retardation candidate gene CDKL3 regulates neuronal morphogenesis.

Mental retardation is a common neurological disorder characterized by various clinical manifestations, primarily impaired cognitive function. To date, only a few genes linked to mental retardation have been well characterized, and the genetics of mental retardation remains poorly understood. Here, we investigated the role of the Cdkl3, a mental retardation candidate gene, in neuronal morphogenesis. We show that Cdkl3 mRNA expression is developmentally regulated in the central nervous system, peaking during neonatal stages, and CDKL3 protein is enriched in neuronal nuclei. Downregulating CDKL3 by RNAi decreased dendrite branching, total dendritic length and complexity in cultured cortical neurons, whereas it promoted axon growth without affecting axon/dendrite specification. Furthermore, depleting CDKL3 in developing cortical neurons also inhibited dendritic growth and maturation, and reduced spine density on basal dendrites in vivo. In contrast, overexpressing CDKL3 enhanced dendritic elaboration both in vitro and in vivo and suppressed axonal outgrowth in vitro. Taken together, these findings demonstrate that CDKL3 is involved in neuronal morphogenesis during development.