LncRNA nuclear-enriched abundant transcript 1 aggravates cerebral ischemia/reperfusion injury through activating early growth response-1/RNA binding motif protein 25 axis.

Ischemic stroke is a major global health issue. Ischemia and subsequent reperfusion results in stroke-related brain injury. Previous studies have demonstrated that nuclear-enriched abundant transcript 1 (NEATa and early growth response 1 (EGR1) are involved in ischemia reperfusion (IR) injury). In this study, we aimed to explore the roles of NEAT1/EGR1 axis as well as its downstream effector RNA binding motif protein 25 (RBM25) in cerebral IR injury. Oxygen-glucose deprivation/reperfusion (OGD/R) and middle cerebral artery occlusion (MCAO) were used to establish in vitro and in vivo models of cerebral IR injury, respectively. According to our data, NEAT1, EGR1, and RBM25 levels were elevated in OGD/R-exposed SK-N-SH and SH-SY5Y cells and cerebral cortex of MCAO mice. NEAT1, EGR1, or RBM25 knockdown effectively reduced infarct volumes and apoptosis, and improved neurological function. Mechanistically, NEAT1 directly interacted with EGR1, which restrained WW domain containing E3 ubiquitin protein ligase 1 (WWP1)-mediated ubiquitination of EGR1 and subsequently caused EGR1 accumulation. EGR1 bound to RBM25 promoter and transcriptionally activated RBM25. Rescue experiments indicated that RBM25 overexpression abolished the therapeutic effects of NEAT1 knockdown. In conclusion, this work identified a novel NEAT1/EGR1/RBM25 axis in potentiating brain injury after IR insults, suggesting a potential therapeutic target for ischemic stroke.

Investigation on Potential Biomarkers and Immune-Related Mechanisms of Ischemic Stroke.

BACKGROUND: This study aimed to reveal the detailed immune-related mechanisms underlying ischemic stroke (IS) and identify new immune-associated biomarkers for clinical management. METHODS: Differentially expressed genes (DEGs) between IS samples and normal controls were identified using the GSE16561 dataset. The feature genes of the immune cells were investigated using the GSE72642 dataset. Weighted correlation network analysis (WGCNA) was performed to reveal module genes, followed by an investigation of common DEGs and a functional enrichment analysis. Potential biomarkers were identified based on hub genes in protein-protein interaction networks and WGCNA. Finally, GSE158312 was used for biomarker verification. RESULTS: In total, 1230 DEGs were identified between the IS samples and normal controls. Seven clinically significant modules were identified using WGCNA. The yellow module genes were positively correlated with polymorphonuclear cells (PMNC), whereas the brown module genes were positively correlated with CD4+ T cells. Eight genes were selected as hub genes. These genes are mainly involved in functions such as the innate immune response. Upregulated TLR2 and ARG1 levels were significantly different between the two groups in the verification dataset. CONCLUSIONS: Our findings suggest ARG1 and TLR2 as novel biomarkers for IS. Upregulated TLR2 might play a role in IS development by participating in the innate immune response function.

Effect of MicroRNA-126a-3p on Bone Marrow Mesenchymal Stem Cells Repairing Blood-brain Barrier and Nerve Injury after Intracerebral Hemorrhage.

OBJECTIVE: Intracerebral hemorrhage (ICH) is a disease that threatens human health due to its high morbidity and mortality. On behalf of finding the better methods in the treatment of ICH, researchers pay more attention to a new technology which is finding effective genes to modify stem cells. METHODS: In this study, we isolated, cultured and identified bone marrow mesenchymal stem cells (MSCs) in vitro. Further, the MSCs (transfected with lentivirus expressing microRNA-126a-3p (miR-126)) were injected into the type Ⅶ collagenase-induced ICH rats to investigate the recovery effects of blood-brain barrier (BBB) and nerve damage in vivo. RESULTS: The MSCs surface marker molecules (CD29: 98.5%; CD90: 96.5%) were highly expressed, and the blood cell surface molecule was negatively expressed (CD45: 2%). Meanwhile, it was verified that miR-126 facilitated the differentiation of MSCs into vascular endothelial cells, owing to the rise of markers (CD31 and VE-cadherin). The modified neurological severity score, modified limb placing test score, brain water content and evans blue content were reduced after transplanted miR-126-modified MSCs. It was found that miR-126 accelerated the differentiation of MSCs into vascular endothelial cells via immunohistochemical staining in vivo. HE staining indicated the area of edema was obviously decreased compared with that in ICH + vector-MSCs group. MiR-126-modified MSCs alleviated the cell apoptosis in brain tissues by TUNEL assay. In addition, the mRNA and protein expression of protease activated receptor-1 and matrix metalloproteinase-9 were diminished, whilst the expression of zonula occludens-1 (ZO-1) and claudin-5 were enhanced in ICH+miR-126-MSCs group. Immunofluorescence assay revealed that miR-126-modified MSCs decreased the disruption of tight junction (ZO-1 and claudin-5). CONCLUSIONS: All data illustrate that miR-126-modified MSCs repair BBB and nerve injury after ICH.

Molecular differences in Alzheimer's disease between male and female patients determined by integrative network analysis.

Alzheimer's disease (AD) is a complex neurodegenerative disease and the most common cause of dementia among the elderly. There has been increasing recognition of sex differences in AD prevalence, clinical manifestation, disease course and prognosis. However, there have been few studies on the molecular mechanism underlying these differences. To address this issue, we carried out global gene expression and integrative network analyses based on expression profiles (GSE84422) across 17 cortical regions of 125 individuals with AD. There were few genes that were differentially expressed across the 17 regions between the two sexes, with only four (encoding glutamate metabotropic receptor 2, oestrogen-related receptor beta, kinesin family member 26B, and aspartoacylase) that were differentially expressed in three regions. A pan-cortical brain region co-expression network analysis identified pathways and genes (eg, glycogen synthase kinase 3ß) that were significantly associated with clinical characteristics of AD (such as neurofibrillary score) in males only. Similarity analyses between region-specific networks indicated that male patients exhibited greater variability, especially in the superior parietal lobule, dorsolateral prefrontal cortex and occipital visual cortex. A network module analysis revealed an association between clinical traits and crosstalk of sex-specific modules. An examination of temporal and spatial patterns of sex differences in AD showed that molecular networks were more conserved in females than in males in different cortical regions and at different AD stages. These findings provide insight into critical molecular pathways governing sex differences in AD pathology.

Olfactory Impairment and Hippocampal Volume in a Chinese MCI Clinical Sample.

PURPOSE: The aim of this study was to evaluate the relationship between olfactory function and hippocampal volume in patients with mild cognitive impairment (MCI). METHODS: We enrolled a total of 31 MCI patients and 9 normal control subjects. All participants underwent 3.0 T-magnetic resonance imaging scanning. The scan results were processed using GE ADW4.6 processing software and V0xar 3D workstation to acquire the hippocampal volume. The University of Pennsylvania Smell Identification Test (UPSIT) was used to evaluate the olfactory function of MCI patients. The correlations of UPSIT score with hippocampal volume and hippocampal head volume were evaluated by Pearson correlation coefficient analysis. RESULTS: MCI patients had significantly smaller left (2.78±0.50 vs. 3.19±0.31 cm(3)) and right (2.97±0.42 vs. 3.31±0.25 cm(3)) hippocampal volumes compared with normal controls (P<0.05). In addition, patients with olfactory dysfunction had smaller volumes of the hippocampus (left hippocampal volume, 2.57±0.39 vs. 3.23±0.40 cm(3); right hippocampal volume, 2.86±0.43 vs. 3.22±0.30 cm(3)) and hippocampal head (left hippocampal head volume, 1.18±0.16 vs. 1.53±0.25 cm(3); right hippocampal head volume, 1.25±0.22 vs. 1.54±0.22 cm(3)) compared with those with normal olfactory function (P<0.05). No significant difference in the hippocampal body volume and hippocampal tail volume was found between MCI patients with olfactory loss and those with normal olfactory function. The UPSIT score was significantly positively correlated with left hippocampal volume (r=0.55, P<0.05), right hippocampal volume (r=0.42, P<0.05), left hippocampal head volume (r=0.53, P<0.05), and right hippocampal head volume (r=0.45, P<0.05). CONCLUSIONS: Olfactory function correlates well with hippocampal volume among patients with MCI.

Identification of pivotal markers in vascular dementia based on proteomics data.

OBJECTIVE: The aim of this study was to analyze protein expression profiles of vascular dementia (VaD) subjects for investigating the underlying therapeutic markers. METHODS: Protein expression profile data were acquired from a quantitative clinical proteomic study, including 10 nondemented elderly controls and 10 age-matched VaD subjects. Differentially expressed proteins (DEPs) were identified between VaD subjects and controls, followed by function prediction using DAVID (Database for Annotation, Visualization, and Integrated Discovery). Then, a protein-protein interaction (PPI) network was constructed by comparing it with the STRING (Search Tool for the Retrieval of Interacting Genes) database, and the pathway crosstalk analysis was conducted based on overlapping PPI network and enriched pathways. Furthermore, the subpathway was screened and analyzed by the iSubpathwayMiner package in R. RESULTS: A total of 144 DEPs were screened from VaD subjects and the controls. They were significantly enriched in many pathways. High-degree proteins were detected in the PPI network, such as ATP5B (ATP synthase subunit ß). Furthermore, 'metabolic pathways' and 'Alzheimer's disease' were the significant pathways screened in the crosstalk analysis. At last, upregulated proteins were enriched in 2 subpathways of 1 pathway, while downregulated proteins were enriched in 162 subpathways of 36 pathways. CONCLUSION: By analyzing the differential expressions of proteins, the potential underlying therapeutic markers and mechanism of VaD might be elucidated.

Vitamin D receptor gene polymorphisms and the risk of Parkinson's disease.

The effect of vitamin D receptor (VDR) gene polymorphisms on Parkinson's disease (PD) has recently gained interest. However, evidence on this relationship is controversial. We searched PubMed, EMBASE and the Cochrane Library database targeted all studies that evaluated VDR gene polymorphisms and PD up to April 2,014. A meta-analysis was conducted on the association between VDR ApaI, BsmI, TaqI and FokI polymorphisms and PD using (1) allelic contrast, (2) dominant, (3) recessive, and (4) additive models. A total of five relevant studies involving PD patients (n = 1,266) and controls (n = 1,649) were included in the analysis. There was a significant association between FokI polymorphism and PD. In the pooled allelic analysis, the F allele was associated with increased risk of PD (OR 1.41, 95% CI 1.14-1.75). In the genotype analysis, FF + Ff versus ff showed a significant association with PD in the dominant model (OR 2.32, 95% CI 1.49-3.61, P = 0.0002). FF versus ff showed a significant association with PD in the additive model (OR 2.45, 95% CI 1.52-3.93, P = 0.0002). There was also a statistically significant association between VDR BsmI polymorphisms in the recessive model, BB versus Bb + bb showed a significant increased risk of PD (OR 1.37, 95% CI 1.01-1.87, P = 0.04). No significant associations were observed between VDR ApaI and TaqI polymorphisms and PD. To sum up, VDR BsmI and FokI polymorphisms were associated with increased risk of PD.

Effect of focal mild hypothermia on expression of MMP-9, TIMP-1, Tau-1 and ß-APP in rats with cerebral ischaemia/reperfusion injury.

PRIMARY OBJECTIVE: Following stroke, hypothermia is reported to reduce both cellular and extracellular damage. This study aimed to examine the effects of focal mild hypothermia on proteins associated with both extracellular (matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of MMP-9 (TIMP-1)) and cellular damage (Tau-1 and ß-amyloid precursor protein (ß-APP)) to characterize the protective effects of hypothermia. METHODS AND PROCEDURES: Male Wistar rats received ischaemic damage using a transient, focal ischaemia/reperfusion model. Afterwards, one group (HT) received 6 hours of focal mild hypothermia (33 °C) applied to the head, while another remained at normal temperature (NT). The brains were collected at 6, 12, 24, 48 and 72 hours after hypothermia to measure infarct volume ratio and to detect cells immunopositive for MMP-9, TIMP-1, Tau-1 and ß-APP, while neurological deficits were examined separately after 2 weeks. MAIN OUTCOMES AND RESULTS: Focal mild hypothermia had no effect on infarct volume ratio but expression of MMP-9, TIMP-1 Tau-1 and ß-APP was decreased. Furthermore, neurological function in the HT group was better than in the NT group. CONCLUSIONS: Focal mild hypothermia has protective effects on cerebral ischaemia-reperfusion injury characterized by decreased expression of MMP-9, TIMP-1, Tau-1 and ß-APP, along with improvement of neurological function despite no changes in infarct volume.

An interventional study of baicalin on neuronal pentraxin-1, neuronal pentraxin-2, and C-reactive protein in Alzheimer's disease rat model.

Background: Baicalin has been shown to promote spatial learning and neural regeneration, which might increase the differentiation of neural stem cells in Alzheimer's disease (AD) rat models. We aimed to study the role of baicalin on neuronal pentraxin-1 (NPTX-1), neuronal pentraxin-2 (NPTX-2), and C-reactive protein (CRP) in AD model rats. Methods: The 30 male Sprague Dawley rats were divided into three groups: the control group, the AD model group, and the AD + baicalin group. Then, the Morris water maze was used to verify the effect of baicalin on the memory and spatial learning of rats. Immunohistochemistry and immunofluorescence were used to observe the expression of NPTX-1, NPTX-2, and CRP in brain tissue. Results: Compared with the AD model group, the AD rats treated with baicalin spent significantly less time finding escape latencies (P = 0.008) and had longer cross-platform times in the target quadrant (P = 0.015). In addition, the AD + baicalin group had significantly higher numbers of hippocampal neurons compared with the AD model group (P < 0.05). Baicalin also obviously decreased the apoptosis of neurons. Moreover, compared with the AD model group, the NPTX-1 and CRP expression in the AD + baicalin group was significantly reduced (P = 0.000) while the expression of NPTX-2 in the brain tissue of AD rats was significantly increased (P = 0.000). Conclusions: Baicalin can play a therapeutic role by downregulating NPTX-1, upregulating NPTX-2, and downregulating CPR in AD model rats.

KDM1A-mediated upregulation of METTL3 ameliorates Alzheimer's disease via enhancing autophagic clearance of p-Tau through m6A-dependent regulation of STUB1.

BACKGROUND: Alzheimer's disease (AD) is a severe neurodegenerative disorder that progressively destroys cognitive skills. Exploring the mechanism underlying autophagic clearance of phosphorylated tau (p-Tau) contributes to developing novel therapeutic strategies for AD. METHODS: SH-SY5Y and HT22 cells were treated with Aß1-42 to establish an in vitro model of AD. Cell viability was examined using CCK-8. TUNEL staining was applied to evaluate cell apoptosis. LC3 puncta was examined by IF staining. m6A modification level was evaluated through MeRIP. RNA pull-down and RIP assays were used for analyzing the interaction between IGF2BP1 and STUB1 transcripts. The binding of KDM1A to the promoter of METTL3 was confirmed by ChIP assays. APP/PS1 transgenic mice were used as an in vivo model of AD. Cognitive skills of mice were evaluated with the Morris water maze. Hippocampal damage and Aß deposition were detected through H&E and IHC staining. RESULTS: Dysregulated levels of autophagy, p-Tau and m6A was observed in an in vitro model of AD. Overexpression of METTL3 or STUB1 enhanced autophagy but reduced p-Tau level in Aß1-42-treated cells. METTL3 stabilized STUB1 mRNA through the m6A-IGF2BP1-dependent mechanism and naturally promoted STUB1 expression, thereby enhancing autophagic p-Tau clearance in Aß1-42-treated cells. Overexpression of KDM1A enhanced autophagy, m6A modification and autophagic p-Tau clearance in Aß1-42-treated cells. KDM1A-mediated upregulation of METTL3 promoted autophagic p-Tau clearance and ameliorated Alzheimer's disease both in vitro and in vivo. CONCLUSION: KDM1A-mediated upregulation of METTL3 enhances autophagic clearance of p-Tau through m6A-dependent regulation of STUB1, thus ameliorating Alzheimer's disease. Our study provides novel mechanistic insights into AD pathogenesis and potential drug targets for AD.

High expression of ubiquitin conjugates and NF-κB in unstable human intracranial atherosclerotic plaques.

Arteries from the first segment of the middle cerebral and adjacent normal blood vessel specimens were collected from 19 patients who had died from cerebral infarction. According to morphologic examination the atherosclerotic plaque present was classified as stable or unstable. The expression of ubiquitin conjugates and nuclear factor kappa B (NF-κB) was assessed and found significantly higher in plaques than normal tissue and higher in unstable plaques than in stable plaques. Ubiquitin conjugates and NF-κB were positively correlated with each other. It was concluded that the ubiquitin proteasome system and NF-κB possibly play a role in the development of atherosclerotic plaques in intracranial arteries.

lncRNA MIR600HG Knockdown Alleviates Cognitive Impairment in Alzheimer's Disease Through NEDD4L Mediated PINK1 Degradation.

BACKGROUND: Growing evidence has demonstrated that long non-coding RNAs (lncRNAs) play a critical role in Alzheimer's disease (AD), which is characterized by sustained mitochondrial dysfunction, inevitable memory loss, and cognitive decline. However, the potential function of lncRNAs MIR600 Host Gene (MIR600HG) in AD remains unanswered. OBJECTIVE: Our study aimed to investigate the role of MIR600HG and its related molecular mechanism in AD. METHODS: The expression of MIR600HG was examined by qRT-PCR. The MIR600HG interacting proteins were identified by RNA pull-down assay and mass spectrometry and verified by RNA immunoprecipitation. Immunofluorescence staining was applied to examine the colocalization of PINK1 and NEDD4L. The PINK1 level and the activation of autophagy were detected by immunoblotting. Morris water maze test was performed to evaluate cognitive decline in AD mice model. RESULTS: MIR600HG expression was elevated during aging in two different types of AD transgenic mouse models. Next, we found that increased MIR600HG directly interact with NEDD4L, which promoted PINK1 ubiquitination and degradation, and as well as autophagy activation. Additionally, MIR600HG promoted Aß production and suppressed Cytochrome C Oxidase activity. Administration of AAV-shMIR600HG restored the Cytochrome C Oxidase activity and inhibited Aß production. Furthermore, PINK1 overexpression or MIR600HG knockdown significantly ameliorated the cognitive impairment in APP/PS1 mice. PINK1 depletion recovered the spatial memory defect in the AAV-shMIR600HG injected APP/PS1 mice. CONCLUSION: MIR600HG was increased in AD and promoted AD pathogenesis. Targeting MIR600HG significantly improved cognitive function in AD mice, which could pave the way for exciting new avenues in AD therapeutic strategy research.

Acute disseminated encephalomyelitis associated with Influenza A H1N1 infection.

Previous studies had not suggested acute disseminated encephalomyelitis (ADEM) during Influenza A H1N1 infection. We report the case of patient who had predominant neurological complication following Influenza A H1N1 infection. The patient, who showed clinical and MRI evidence of ADEM, had significant recovery, which in part, may be related to early treatment. The patient demonstrated that the prognosis of Influenza A H1N1-associated ADEM may not be poor.

Protective effect of edaravone against PrP106-126-induced PC12 cell death.

The prion protein peptide PrP106-126 induces cell apoptosis through mechanisms involving production of intracellular reactive oxygen species. The present study investigated the effects of edaravone, a potent free radical scavenger in clinical use, on cell cytotoxicity induced by PrP106-126. Results showed that PrP106-126 decreased PC12 cell viability in a dose- and time-dependent manner. Edaravone significantly antagonized the cytotoxic effects of PrP106-126. Mechanistically, PrP106-126 decreased PC 12 intracellular glutathione (GSH) concentrations, decreased superoxide dismutase (SOD) enzyme activity, increased concentrations of the oxidation end product malondialdehyde (MDA), depolarized the mitochondrial membrane, and increased caspase-3 activity. Edaravone alone did not affect GSH, SOD, or MDA but did effectively reverse all of the intracellular prooxidant effects induced by PrP106-126 and inhibit induced apoptosis in PC12 cells. In conclusion, edaravone may be a viable candidate for the treatment of oxidative stress-induced neurodegenerative disease.

Perihematomal pathological changes in neurons and astrocytes following acute cerebral hemorrhage.

We aim to investigate the pathological temporospatial characteristics of brain cell injury in the perihematomal areas. Brain autopsy samples from 44 consecutive cases of intracerebral hemorrhage were processed and analyzed following immunohistochemical staining for neurofilament (NF) and glial fibrillary acidic protein (GFAP). NF and GFAP positive cells were scored and graded according to the distance from the hematoma and the time from the onset of hematoma formation. The tissues from the same region on the contralateral side of the brain were used as controls. Neurons in the perihematomal areas exhibited pyknosis or swollen necrosis, while astrocytes were swollen. Morphological abnormalities pertaining to NF appearance were attenuated with increasing distance from the hematoma wall, but were exacerbated with prolonged bleeding time. The level of NF staining abnormality was positively correlated with time from the onset of hematoma within 7 days of intracerebral hemorrhage. In contrast, the intensity of GFAP staining was negatively correlated with time from the onset of hematoma formation. This immunoreactivity was significantly higher closer to hematoma. Taken together, these data indicate that pathological alterations in neurons and astrocytes in the perihematomal area change with time from the onset of hematoma formation.

Exosomes Derived from MicroRNA-146a-5p-Enriched Bone Marrow Mesenchymal Stem Cells Alleviate Intracerebral Hemorrhage by Inhibiting Neuronal Apoptosis and Microglial M1 Polarization.

INTRODUCTION: Intracerebral hemorrhage (ICH) is a devastating type of stroke with high mortality, and the effective therapies for ICH remain to be explored. Exosomes (Exos) have been found to play important roles in cell communication by transferring molecules, including microRNAs (miRNAs/miRs). MiRNAs are critical regulators of genes involved in many various biological processes and have been demonstrated to aggravate or alleviate brain damages induced by ICH. The aim of the present study was to investigate the effect of Exos derived from miR-146a-5p-enriched bone marrow mesenchymal stem cells (BMSCs-miR-146a-5p-Exos) on experimental ICH. METHODS: ICH was induced in adult male Sprague-Dawley rats by an intrastriatal injection of collagenase type IV. At 24 h after surgery, Exos were administrated. For detecting apoptotic cells, TUNEL staining was performed using an in situ Cell Death Detection Kit. Fluoro-Jade B staining was performed to detect degenerating neurons. Immunofluorescence assay was performed to detect the expression of myeloperoxidase (MPO) and OX-42. The binding of miR-146a-5p and its target genes was confirmed by luciferase reporter assay. RESULTS: At 24 h after surgery, BMSCs-miR-146a-5p-Exos administration significantly improved neurological function, reduced apoptotic and degenerative neurons, and inhibited inflammatory response. Furthermore, miR-146a-5p-enriched Exos obviously inhibited the M1 polarization of microglia after ICH in rats, accompanied by the reduced expression of pro-inflammatory mediators releasing by M1 microglia including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and monocyte chemoattractant protein-1 (MCP-1). Finally, we observed that miR-146a-5p directly targeted interleukin-1 receptor-associated kinase1 (IRAK1) and nuclear factor of activated T cells 5 (NFAT5), which contributed to the inflammation response and the polarization of M1 microglia/macrophages. CONCLUSION: We demonstrated that miR-146a-5p-riched BMSCs-Exos could offer neuroprotection and functional improvements after ICH through reducing neuronal apoptosis, and inflammation associated with the inhibition of microglial M1 polarization by downregulating the expression of IRAK1 and NFAT5.

Analysis of fuzhisan and quantitation of baicalin and ginsenoside Rb(1) by HPLC-DAD-ELSD.

Fuzhisan (FZS) is a traditional Chinese medicine composed of Radix Ginseng Rubra (Araliaceae family), Scutellaria baicalensis Georgi (Labiatae family), Angelica sinensis (Oliv) Diels (Umbelliferae family), Anemone altaica Fisch. Ex C.A. Mey (Araceae family) and Glycyrrhiza uralensis (Leguminosae family). To establish the chemical fingerprint of the components of FZS and quantify the components, baicalin and ginsenoside Rb(1), a high performance liquid chromatography method coupled with diode array and evaporative light scattering detectors (DAD-ELSD) was developed. Separation of 36 components from 12 batches of FZS was performed on a C(18) column, with a mobile phase consisting of acetonitrile and 0.1% acetic acid-water, with gradient elution at a column temperature of 30 degrees C. The optimum detection wavelength was set at 335 nm, the drift tube temperature of ELSD was set at 80 degrees C, the carrier gas pressure was 25 psi, and the gain = 10. The similarity among 12 batches of FZS was over 0.95. Five constituents of FZS, namely baicalin, ferulic acid, and ginsenosides Rg(1), Re, and Rb(1), were identified based on their retention times (RT). Calibration curves for baicalin and ginsenoside Rb1 showed good linearity (r (2) > 0.9992); recoveries ranged from 95% to 99%. This quantification method is reproducible and simple, and may provide a tool to assess the quality of FZS.

[Influences of bFGF and EGF on the proliferation and differentiation of endogenous neural stem cells after cerebral infarction in human].

OBJECTIVE: To investigate the relationship of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) and proliferation of endogenous neural stem cells (NSCs) after human cerebral infarction. METHODS: Paraffin-embedded brain tissues of 22 human fatal cases of CI from the brain tissues around subventricular zone and subgranular layer zone were stained with HE and immunohistochemistry stain. The endogenous neural stem cells were marked by nestin. The expression changes of EGF, bFGF and nestin in the perihematomal tissues were analysed with the SPSS 13.0 system. RESULTS: (1) Compared with the controls, the number of nestin-positive cells increased at 24 - 72 h (14 +/- 6)/HP in the ipsilateral SVZ and began to rise at 4.5 - 10 h (11 +/- 5)/HP in the ipsilateral SGZ, reached maximum at 120 - 144 h ((38 +/- 7)/HP in the SVZ, (54 +/- 17)/HP in the SGZ, and decreased markedly at 216 - 336 h, but it was still elevated compared with the controls (P < 0.05). (2) The number of bFGF-positive cells increased at 4.5 - 10 h (8.1 +/- 2.9)/HP in the SVZ, (19.0 +/- 8.2)/HP in the SGZ, reached maximum at 24 - 70 h (15.6 +/- 3.5)/HP in the SVZ, (32.0 +/- 5.7)/HP in the SGZ and decreased at 72 - 96 h, but it was still elevated compared with the controls (P < 0.05). (3) The number of EGF-positive cells increased at 4.5 - 10 h (4.3 +/- 1.6)/HP in the SVZ, (7.0 +/- 3.7)/HP in the SGZ, reached maximum at 120 - 144 h (27.0 +/- 1.4)/HP in the SVZ, (51.5 +/- 4.9)/HP in the SGZ and decreased at 216 - 336 h, but it was still elevated compared with the controls (P < 0.05). CONCLUSIONS: Perhaps the increased expression of EGF and bFGF after CI was a reaction of endogenous reparation and it correlated with the proliferation and endogenous of neural stem cells in human.

Baicalin and ginsenoside Rb1 promote the proliferation and differentiation of neural stem cells in Alzheimer's disease model rats.

BACKGROUND: This study aimed to explore the effects of ginsenoside Rb1 and baicalin on the proliferation and differentiation of neural stem cells (NSC) in Alzheimer's disease model rats. METHOD: The healthy Sprague Dawley male rats were randomly divided into 4 groups: control group, model group, ginsenoside Rb1 group and baicalin group. Besides, the animal model of dementia was induced by the injection of Aß1-40. 2 weeks later, the rats in the baicalin and ginsenoside Rb1 groups were injected with baicalin and ginsenoside Rb1, respectively. The contents, expression sites of Nestin, GFAP and NSE and the percentage of viable cells were detected by immunohistochemistry. In addition, the expression levels of Nestin, GFAP and NSE in hippocampus of rats were detected by western-blot and metrology analysis was performed using quantity. RESULTS: Injection of Aß1-40 significantly reduced the number of neuronal cells (p < .05). In addition, compared with the control group, the percentages of positive cells of NSCs, astrocytes and neuronal were increased. Besides, compared with the model group, the percentage of positive neural cells was improved by ginsenoside Rb1 (p < .05), and the percentages of astrocytes and neuronal were increased by ginsenoside Rb1 and baicalin (p < .05). Moreover, the expressions of Nestin and NSE were enhanced by ginsenoside Rb1 and baicalin (p < .05), while the GFAP level was only affected by ginsenoside Rb1 (p < .05) when compared with the model group. CONCLUSION: Ginsenoside Rb1 and baicalin might promote the proliferation and differentiation of endogenous NSCs in AD rat model.

[Expressions of heme oxygenase-1 and apoptosis-modulating proteins in peri-hematoma cortex after intracerebral hemorrhage in human being].

OBJECTIVE: To observe the expression of heme oxygenase-1 (HO-1) and Bcl-2, an apoptosis-modulating protein, in the neurons surrounding the hematoma in human being. METHODS: Specimens of cerebral cortex tissue 1 - 3 cm around the hemorrhagic focus with the size of 2.0 cm x 1.5 cm x 0.3 cm were collected during autopsy from 39 patients, 17 males and 22 females, aged 62.8 (36 - 84), who died from intracerebral hemorrhage 2 - 10 h, 17 - 30 h, 36 - 96 h, 120 - 216 h, or 240 - 408 h before. Specimens of brain tissue of the same size at the opposite side were collected as controls. Immunohistochemistry was used to examine the expression of HO-1 and Bcl-2 protein. RESULTS: (1) Expression of HO-1 could be detected in the specimens of the 2 h group, increased in the specimens of the 2 - 10 h group [(5.1 +/- 2.0)/HP], reached the peak in the 17 - 30 h group [(11.3 +/- 0.9)/HP], then began to decrease in the specimens of the 240 - 408 h group [(6.4 +/- 0.6)/HP] (F = 42.80, P < 0.001). The HO-1 expression of the control group remained negative at any time-point. (2) Expression of Bcl-2 could be detected in the specimens of the 2 - 10 h group [(4.2 +/- 1.7)/HP], was increased in the 17 - 30 h group [(6.6 +/- 0.5)/HP], reached the peak in the 36 approximately 96 h group [(8.9 +/- 1.1)/HP], then began to decrease, and was (4.7 +/- 0.6)/HP in the 240 approximately 408 h group (F = 29.59, P < 0.001). The Bcl-2 expression remained negative at any time point in the control group. (3) The expressions of HO-1 was positively correlated with the expression of Bcl-2 (r = 0.66, P < 0.001). CONCLUSIONS: Over-expression of HO-1 and Bcl-2 in the neurons provide a potential protection or destruction mechanism after intracerebral hemorrhage in human.