Haiwei Meng

Effects of Ginkgolide B on 6-OHDA-induced apoptosis and calcium over load in cultured PC12

Ginkgolide B, one of the major components of Ginkgo biloba extracts, is a potent platelet‐activating factor (PAF) receptor antagonist, which is also regarded as having neuroprotective effects on the CNS. The aim of this research is to observe the effects of Ginkgolide B on the PC12 apoptosis induced by 6‐hydroxydopamine (6‐OHDA) and to explore whether these effects are related to the changes of intracellular Ca2+ and Calbindin D28K mRNA in PC12 cells. In the present work, the damage of PC12 cells was induced by 100 μM 6‐OHDA. The cells survival rate was examined by MTT assays. The intracellular free calcium concentration in PC12 cells was measured by using the fluorescent Ca2+ indicator fluo‐3/AM. Semi‐quantitative reverse transcriptase‐polymerase chain reaction (RT‐PCR) was employed to determine the expression of Calbindin D28K mRNA in PC12. The data show that the Ginkgolide B inhibited PC12 cells apoptosis induced by 6‐OHDA in a dose‐dependent manner, and decreased the activity of caspase‐3. In addition, Ginkgolide B increased the expression of Calbindin D28K mRNA and inhibited 6‐OHDA‐induced elevation in the intracellular calcium concentration. Our results showed that the Ginkgolide B inhibited the apoptosis of PC12 induced by 6‐OHDA, and the protective effects of Ginkgolide B on PC12 cells are mediated, at least in part, by up‐regulating the Calbindin D28K mRNA and by decreasing the intracellular calcium concentration.

D-β-hydroxybutyrate inhibits the apoptosis of PC12 cells induced by 6-OHDA in relation to up-regulating the ratio of Bcl-2/Bax mRNA

D-beta-hydroxybutyrate (DbetaHB) is a predominant member of ketone bodies produced by hepatocytes and, to a lesser extent, by astrocytes. It is an alternative source of energy in the brain when glucose supply is depleted such as during starvation. It has been reported that ketone bodies could protect dopaminergic culture. However, the biological function of DbetaHB in Parkinson disease (PD) is still unclear. In the present work, we investigated the role of DbetaHB in protecting rat pheochromocytoma (PC12) cells from apoptosis induced by 6-Hydroxydopamine (6-OHDA). DbetaHB rescued PC12 cells from apoptotic death induced by 6-OHDA by MTT assay, acridine orange (AO) staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining and the activity of caspase-3. DbetaHB prevented the decrease of cell viability and the increase of caspase-3 activity induced by 6-OHDA in a dose-dependent manner in PC12 cells. AO and TUNEL staining showed that DbetaHB prevented the apoptosis of PC12 cells induced by 6-OHDA. The ratio of Bcl-2/Bax at mRNA levels, which regulates the apoptosis of PC12 cells when exposed to 6-OHDA, increased when DbetaHB was preincubated. The data showed that DbetaHB inhibited the apoptosis of PC12 cells induced by 6-OHDA in relation to up-regulating the ratio of Bcl-2/Bax mRNA.

α-Synuclein knockdown attenuates MPP + induced mitochondrial dysfunction of SH-SY5Y cells

Alpha-synuclein is one of the main constituents of Lewy bodies and plays an important role in the pathology of Parkinsons disease. Mutation or overexpression of alpha-synuclein causes Parkinsons disease, and downregulation of alpha-synuclein resists MPP(+)-induced cell death, but the mechanism remains elusive. In this study, we attempted to explore the effect of alpha-synuclein knockdown on mitochondrial function in MPP(+)-treated SH-SY5Y cells. We reconstructed the short hairpin RNA expression vector, pGenesil-2, specially targeting alpha-synuclein mRNA, and it was stably transfected into SH-SY5Y cells. Cell viability, nuclear morphology, and mitochondrial membrane potential were then detected, and the expression of alpha-synuclein, cytochrome c, Bcl-2 and Bax were analyzed by Western blotting. The results showed that after exposure to 500 microM MPP(+) for 24 h, about 41.0+/-1.5% control cells showed low mitochondrial membrane potential. However, the percentage was 13.6+/-1.2% in MPP(+) treated alpha-synuclein knockdown cells. MPP(+) induced cytochrome c release significantly, which was about 3.1-fold compared with that of control. However, in alpha-synuclein knockdown cells, the release of cytochrome c was blocked, which was about 1.4-fold compared with that of control. The Bcl-2/Bax ratio of SH-SY5Y cells reduced to 35.5+/-3.8% after MPP(+) treatment, and this ratio was 85.2+/-3.0% in MPP(+) treated alpha-synuclein knockdown cells. These data suggest that knockdown of alpha- synuclein might be an effective means in rescuing MPP(+)-induced mitochondrial dysfunction of SH-SY5Y cells.

Fiber Pathways of Attention Subnetworks Revealed with Tract-Based Spatial Statistics (TBSS) and Probabilistic Tractography

It has been widely accepted that attention can be divided into three subnetworks - alerting, orienting and executive control (EC), and the subnetworks of attention are linked to distinct brain regions. However, the association between specific white matter fibers and the subnetworks of attention is not clear enough. Using diffusion tensor imaging (DTI), the white matter connectivity related to the performance of attention was assessed by attention network test (ANT) in 85 healthy adolescents. Tract-based spatial statistics (TBSS) and probabilistic diffusion tractography analysis demonstrated that cerebellothalamic tract was involved in alerting, while orienting depended upon the superior longitudinal fasciculus (SLF). In addition, EC was under the control of anterior corona radiata (ACR). Our findings suggest that different fiber pathways are involved in the three distinct subnetworks of attention. The current study will yield more precise information about the structural substrates of attention function and may aid the efforts to understand the neurophysiology of several attention disorders.

Development of the Fetal Cerebral Cortex in the Second Trimester: Assessment with 7T Postmortem MR Imaging

BACKGROUND AND PURPOSE: Few investigators have analyzed the fetal cerebral cortex with MR imaging of high magnetic strength. Our purpose was to document the sulcal development and obtain quantitative measurements of the fetal brain in the second trimester. MATERIALS AND METHODS: The brains of 69 fetal specimens, with GA 12–22 weeks, were first scanned on a 7T MR imaging scanner. Then the sequential development of the different fissures and sulci was analyzed, and quantitative measurements of the cerebral cortex were obtained. RESULTS: A new chronology of sulcal development during 12–22 weeks GA was summarized. Before 12 weeks, few sulci were present; by 16 weeks, many sulci were present. The 16th week could be considered the most intensive time point for sulcal emergence. Most sulci, except for the postcentral sulcus and intraparietal sulcus, were present by 22 weeks GA. Measurements of the fetal brains, each with different growth rates, linearly increased with GA, but no sexual dimorphisms or cerebral asymmetries were detected. CONCLUSIONS: The second trimester is the most important phase, during which most sulci are present and can be clearly shown on 7T postmortem MR imaging. It is apparent that the specific time during which neuropathologic features of sulci appear, previously thought to be well understood, should be redefined. Quantitative data provide assistance in the precise understanding of the immature brain. The present results are valuable in anatomic education, research, and assessment of normal brain development in the uterus.

Development of the human fetal cerebellum in the second trimester: a post mortem magnetic resonance imaging evaluation

The cerebellum is one of the most important structures in the posterior cranial fossa, but the characterization of its development by magnetic resonance imaging (MRI) is incomplete. We scanned 40 fetuses that had no morphological brain disorder at 14–22 weeks of gestation using 7.0 T MRI. Amira 4.1 software was used to determine morphological parameters of the fetal cerebellum, which included the cerebellar volume (CV), transverse cerebellar diameter (TCD), and the length and width of the vermis. The relationship between these measurements and gestational age (GA) was analysed. We found that the primary fissure was visible at week 14 of gestation. From week 16, the prepyramidal fissure, the secondary fissure and the dentate nucleus could be identified. The posterolateral fissure and the fourth ventricle were recognized at week 17, whereas the tentorium of the cerebellum was visible at week 20. The relationships between GA and CV, TCD, and the width and length of the vermis were described adequately by second‐order polynomial regression curves. The ratios between TCD and vermis length and between TCD and vermis width decreased with GA. These results show that 7.0 T MRI can show the trajectory of cerebellar development clearly. They increase our understanding of normal cerebellar development in the fetus, and will facilitate the diagnosis of pathological intrauterine changes in the cerebellum.

Cerebral glucose metabolism: Influence on perihematomal edema formation after intracerebral hemorrhage in cat models

Background: Most intracerebral hemorrhage (ICH) imaging studies focus on structural brain changes. Stereotactic neuroimaging techniques such as magnetic resonance imaging (MRI) and positron emission tomography/computed tomography (PET/CT) are helpful in the diagnosis of ICH, monitoring the subsequent recovery and investigating its functional mechanisms. Purpose: To explore the influence of the changes in cerebral glucose metabolism on perihematomal edema formation in an experimental cat model of ICH. Material and Methods: Forty-eight cats were divided into 1 sham operation group (6 cats) and 7 ICH model groups (42 cats)”. The ICH model groups were injected with 1.0 ml autologous nonheparinized blood into their thalami using accurate stereotactic guidance apparatus. MRI and 18F-fluorodeoxyglucose (FDG) PET/CT scans were acquired at 2, 6, 12, 24, 48, 72, and 120 h following the intervention. Pearsons correlation test was used to evaluate the association between T2-weighted signal intensity and the edema volume. Students t test and q test were used to identify the times of significant temporal changes. Results: The volume of perilesional edema did not significantly increase from 2 h to 12 h after ICH, but then increased by 229.4% at 24 h, peaked (by 273.5%), and steadily decreased by 72 h. The FDG intensity in perihematomal edema tissues was markedly reduced 2 h after ICH on PET images, reached its lowest level at 12 h, and then steadily increased at 24 h and 48 h. The changes of standard absorption value (SUV) in perihematomal edema were consistent with those of FDG intensity. Conclusion: Perihematomal glucose metabolism abnormalities have a close relationship with the formation of vasogenic edema. Furthermore, abnormal glucose metabolism may impair capillary integrity and increase blood–brain barrier permeability.

Development of the human fetal hippocampal formation during early second trimester

Development of the fetal hippocampal formation has been difficult to fully describe because of rapid changes in its shape during the fetal period. The aims of this study were to: (1) segment the fetal hippocampal formation using 7.0 T MR images from 41 specimens with gestational ages ranging from 14 to 22 weeks and (2) reveal the developmental course of the fetal hippocampal formation using volume and shape analyses. Differences in hemispheric volume were observed, with the right hippocampi being larger than the left. Absolute volume changes showed a linear increase, while relative volume changes demonstrated an inverted-U shape trend during this period. Together these exhibited a variable developmental rate among different regions of the fetal brain. Different sub-regional growth of the fetal hippocampal formation was specifically observed using shape analysis. The fetal hippocampal formation possessed a prominent medial-lateral bidirectional shape growth pattern during its rotation process. Our results provide additional insight into 3D hippocampal morphology in the assessment of fetal brain development and can be used as a reference for future hippocampal studies.

Rosiglitazone Protects Dopaminergic Neurons Against Lipopolysaccharide-Induced Neurotoxicity Through Inhibition of Microglia Activation

ABSTRACT Recent evidence has suggested that microglia activation plays an important role in the pathogenesis of Parkinsons disease (PD). Activated microglia secrete various proinflammatory cytokines and neurotoxic mediators, which may contribute to the development of PD. Thus, the inhibition of microglia activation may have a therapeutic benefit in the treatment of PD. In the present study, using mesencephalic neuron–microglia mixed culture and microglia-enriched culture, we investigated whether rosiglitazone (RGZ), a member of peroxisome proliferator-activated receptor gamma (PPARγ) agonists, could inhibit microglia activation. Our results showed that RGZ significantly inhibited lipopolysaccharide (LPS)-induced microglia activation and the production of tumor necrosis factor-alpha (TNF-α), nitric oxide (NO), and superoxide. We further investigated the intracellular signaling pathways regulating the production of TNF-α and NO in LPS-activated microglia. The results showed that RGZ inhibited the phosphorylation and nuclear translocation of the p65 subunit of NF-κB, and the phosphorylation of p38 mitogen-activated protein kinase (p38MAPK). Taken together, our results suggested that the therapeutic effects of RGZ were partially mediated by modulating microglia activation.

Measurements using 7.0 T post-mortem magnetic resonance imaging of the scalar dimensions of the fetal brain between 12 and 20 weeks gestational age

In this study, scalar values for the fetal brain from 12 to 20 weeks gestational age were obtained. Fifty‐two fetal specimens of 12–20 weeks gestational age with an anatomically normal and developmentally appropriate central nervous system (CNS) were scanned using a 7.0 T magnetic resonance imaging (MRI) scanner. The linear biometric measurements of the brain were then determined. All the measurements (except for the interhemispheric distance) were found to increase linearly with gestational age, although each increased at a different growth rates. The 95% confidence interval for each value was obtained. These data may be considered to be a valuable reference for the assessment of normal fetal brain development in clinical settings and as a supplement to post‐mortem MRI or anatomical investigations.