MCA供血区急性脑梗死SWI图像与脑血管反应性的相关性

目的:分析大脑中动脉供血区(MCA)急性脑梗死患者的磁敏感加权成像(SWI),探讨梗死侧侧脑室旁深部髓质静脉(DMV)数量与脑血管反应性(CVR)的相关性。方法:回顾性纳入首次发生患侧MCA供血区卒中患者109例,并已完成TCD CO_2负荷实验和SWI等检查。根据SWI图像将DMV数量分为3级,5条为1级;5-10条为2级;10条为3级,分析DMV分级与CVR的关系。结果:根据排除标准共纳入MCA供血区急性脑梗死患者53例,DMV 1级、2级和3级分别有29、18、6例患者,MCA平均CVR为0.40±0.18。DMV 1级CVR为(0.45±0.22)、2级为(0.35±0.15)和3级为(0.20±0.11),3组患者间CVR存在明显差异(P0.05),且DMV 3级组较DMV 1级组、2级组显著降低(P0.05);Spearman相关分析表明,DMV分级与CVR间存在明显的负性相关关系(P0.001)。结论:MCA供血区脑梗死患者DMV分级与患侧MCA CVR呈负相关关系。     

循环血游离Shope病毒DNA含量与兔VX2肿瘤18F-FDG PET/CT影像表现的相关性分析

目的:探讨循环血中Shope病毒DNA含量与兔VX2肿瘤18F-FDG PET/CT影像学特征间的关系及其临床意义。方法:采用组织块接种法建立兔VX2肿瘤模型,并行18F-FDG PET-CT观测肿瘤大小及糖代谢相关值,实时定量荧光探针PCR法检测肿瘤组织及血浆中Shope病毒特征性DNA片段含量。结果:移植前外周血中未检测出Shope病毒特异性DNA片段;移植后2周,VX2肿瘤组织和循环血中均可以检测到特征性Shope病毒DNA片段。瘤体内DNA含量明显高于循环血中含量。循环血Shope病毒DNA含量与FDG-PET的最大标准摄取值(SUVmax)明显呈正相关(r=0.943,p=0.005),但与肿瘤体积相关性尚不明确(r=0.657,p=0.156)。结论:循环血Shope病毒DNA有望作为一种潜在的VX2肿瘤标志物,其廉价、无创的特性,有望在肿瘤的早期诊断和预后随访中发挥优势。     

Functional Imaging of Brown Fat in Mice with 18F-FDG micro-PET/CT

Brown adipose tissue (BAT) differs from white adipose tissue (WAT) by its discrete location and a brown-red color due to rich vascularization and high density of mitochondria. BAT plays a major role in energy expenditure and non-shivering thermogenesis in newborn mammals as well as the adults ¹. BAT-mediated thermogenesis is highly regulated by the sympathetic nervous system, predominantly via β adrenergic receptor ^{2, 3}. Recent studies have shown that BAT activities in human adults are negatively correlated with body mass index (BMI) and other diabetic parameters ^4-6. BAT has thus been proposed as a potential target for anti-obesity/anti-diabetes therapy focusing on modulation of energy balance ^6-8. While several cold challenge-based positron emission tomography (PET) methods are established for detecting human BAT ^9-13, there is essentially no standardized protocol for imaging and quantification of BAT in small animal models such as mice. Here we describe a robust PET/CT imaging method for functional assessment of BAT in mice. Briefly, adult C57BL/6J mice were cold treated under fasting conditions for a duration of 4 hours before they received one dose of ¹⁸F-Fluorodeoxyglucose (FDG). The mice were remained in the cold for one additional hour post FDG injection, and then scanned with a small animal-dedicated micro-PET/CT system. The acquired PET images were co-registered with the CT images for anatomical references and analyzed for FDG uptake in the interscapular BAT area to present BAT activity. This standardized cold-treatment and imaging protocol has been validated through testing BAT activities during pharmacological interventions, for example, the suppressed BAT activation by the treatment of β-adrenoceptor antagonist propranolol ^{14, 15}, or the enhanced BAT activation by β3 agonist BRL37344 ¹⁶. The method described here can be applied to screen for drugs/compounds that modulate BAT activity, or to identify genes/pathways that are involved in BAT development and regulation in various preclinical and basic studies.     

Stereological and Flow Cytometry Characterization of Leukocyte Subpopulations in Models of Transient or Permanent Cerebral Ischemia

Microglia activation, as well as extravasation of haematogenous macrophages and neutrophils, is believed to play a pivotal role in brain injury after stroke. These myeloid cell subpopulations can display different phenotypes and functions and need to be distinguished and characterized to study their regulation and contribution to tissue damage. This protocol provides two different methodologies for brain immune cell characterization: a precise stereological approach and a flow cytometric analysis. The stereological approach is based on the optical fractionator method, which calculates the total number of cells in an area of interest (infarcted brain) estimated by a systematic random sampling. The second characterization approach provides a simple way to isolate brain leukocyte suspensions and to characterize them by flow cytometry, allowing for the characterization of microglia, infiltrated monocytes and neutrophils of the ischemic tissue. In addition, it also details a cerebral ischemia model in mice that exclusively affects brain cortex, generating highly reproducible infarcts with a low rate of mortality, and the procedure for histological brain processing to characterize infarct volume by the Cavalieri method.     

A Method for the In Vivo Investigation of the Serotonergic 5-HT2 Receptors in the Human Cerebral Cortex Using Positron Emission Tomography and 18F-Labeled Setoperone

Following previous validation in baboons, we have studied the characteristics of [18F]setoperone as a radioligand for investigating serotonergic 5-hydroxytryptamine2 (5-HT2) receptors in the normal, unmedicated human brain with positron emission tomography (PET); subjects orally pretreated with therapeutic amounts of ketanserin, sulpiride, or prazosin were also studied to evaluate the specificity and sensitivity of [18F]setoperone brain specific binding. In controls (n = 10), the tracer showed a clear-cut retention in both frontal cortex and striatum (known to contain a high density of 5-HT2 receptors) relative to cerebellum (known to be devoid of 5-HT2 receptors). In the seven young controls (20-39 years old), the frontal cortex/cerebellum and striatum/cerebellum ratios increased during the first hour to reach similar values of 2.53 +/- 0.12 and 2.38 +/- 0.11 (mean +/- SEM), respectively, and were essentially stable during the second hour. Pretreatment with ketanserin (a 5-HT2 blocker) significantly reduced the frontal cortex/cerebellum ratio to 0.7-1.0 at 65 min, whereas the striatum/cerebellum ratio was significantly, but only partially, reduced. During sulpiride treatment (a D2 blocker), the frontal cortex/cerebellum ratio was not altered, whereas the striatum/cerebellum ratio was significantly, but only partially, reduced. With prazosin pretreatment (an alpha 1-adrenergic blocker), neither the frontal cortex/cerebellum nor the striatum/cerebellum ratio was modified. These data in humans with PET demonstrate that [18F]setoperone labels with high sensitivity and selectivity 5-HT2 receptors in the frontal cortex; in striata, however, binding is to both 5-HT2 and D2 receptors. The deproteinated-to-whole plasma radio-activity concentration ratio increased with time following injection. The mean percentage of intact [18F]setoperone, in deproteinated plasma, was 82, 74, 53, 45, 30, and 22% at 5, 10, 20, 30, 60, and 110 min following injection, respectively. These data indicate that [18F]setoperone (a) is significantly bound to plasma proteins and (b) is significantly metabolized into several labeled metabolites that are much more hydrophilic than setoperone and, hence, presumably do not cross the blood-brain barrier. These results suggest the suitability of [18F]setoperone data for modeling of 5-HT2 receptor binding in brain.     

Design,Synthesis, Lipophilic Properties,and Binding Affinities of Potential Ligands in Positron Emission Tomography (PET) for Visualization of Brain Dopamine D4 Receptors

We report the synthesis of compounds structurally related to the high‐affinity dopamine D₄ receptor ligand N‐{2‐[4‐(3‐cyanopyridin‐2‐yl)piperazin‐1‐yl]ethyl}‐3‐methoxybenzamide ( 1e ). All compounds were specifically designed as potential PET radioligands for brain D₄ receptor visualization, having lipophilicity within a range for brain uptake and weak non‐specific binding (0.75<cLogP<3.15) and bearing a substituent for easy access to labeling with the positron emitter isotope ¹¹C or ¹⁸F. The best compound of the series, N‐{2‐[4‐(4‐chlorophenyl)piperazin‐1‐yl]ethyl}‐6‐fluoropyridine‐3‐carboxamide ( 7a ), displayed excellent selectivity over D₂ and D₃ receptors (>100‐fold), but its D₄ receptor affinity was suboptimal for imaging of brain D₄ receptors (K_i=30 nM ).     

Mechanistic Positron Emission Tomography Studies of 6-[18F]Fluorodopamine in Living Baboon Heart: Selective Imaging and Control of Radiotracer Metabolism Using the Deuterium Isotope Effect

Abstract: Mechanistic positron emission tomography (PET) studies using the deuterium isotope effect and specific pharmacological intervention were undertaken to examine the behavior of 6-[¹⁸F]fluorodopamine (6-[¹⁸F]FDA; 1 ) and (?)-6-[¹⁸F]fluoronorepinephrine {(?)-6-[¹⁸F]FNE; 2 } in the baboon heart. Two regiospecifically deuterated derivatives of 6-[¹⁸F]FDA [α,α-D₂(3 ) and β,β-D₂ (4 )] were used to assess the contributions of monoamine oxidase (MAO) and dopamine β-hydroxylase, respectively, to the clearance kinetics of 6-[¹⁸F]FDA. Compound 3 showed a reduced rate of clearance, consistent with MAO-catalyzed cleavage of the α C-D bond, whereas compound 4 showed no change, indicating that cleavage of the β C-D bond is not a rate-limiting step. Pretreatment with pargyline, an MAO inhibitor, also decreased the rate of clearance. Desipramine and tomoxetine [norepinephrine (NE) uptake inhibitors], but not GBR-12909 (a dopamine uptake inhibitor), blocked the uptake of both (?)-6-[¹⁸F]FNE and 6-[¹⁸F]FDA, with (?)-6-[¹⁸F]FNE showing a higher degree of blockade. Chiral HPLC demonstrated that 6-[¹⁸F]FDA is stereoselectively converted to (?)-6-[¹⁸F]FNE in vivo in the rat heart. These studies demonstrate that (a) the more rapid clearance of 6-[¹⁸F]FDA relative to (?)-6-[¹⁸F]FNE can be largely accounted for by metabolism by MAO; (b) selective deuterium substitution can be used to protect a radiotracer from metabolism in vivo and to favor a particular pathway; (c) 6-[¹⁸F]FDA and (?)-6-[¹⁸F]FNE share the NE transporter; (d) 6-[¹⁸F]FDA is stereoselectively converted to (?)-6-[¹⁸F]FNE in vivo; and (e) the profile of radioactivity in the heart for 6-[¹⁸F]FDA is complex, probably including labeled metabolites as well as neuronal and nonneuronal uptake.     

In Vivo Imaging of Vesicular Monoamine Transporters in Human Brain Using [11C]Tetrabenazine and Positron Emission Tomography

Abstract: The pharmacokinetics of [¹¹CJtetrabenazine, a high-affinity radioligand for the monoamine vesicular transporter, were determined in living human brain using in vivo imaging by positron emission tomography (PET). The radiotracer showed high brain uptake and rapid washout from all brain regions with relatively slower clearance from regions of highest concentrations of monoamine vesicular transporters (striatum), resulting in clear differential visualization of these structures at short intervals after injection (10–20 min). As the first human PET imaging study of a vesicular neurotransmitter transporter, these experiments demonstrate that external imaging of vesicular transporters forms a new and valuable approach to the in vivo quantification of monoaminergic neurons, with potential application to the in vivo study of neurodegenerative disorders such as Parkinson's disease.     

Blood–Cerebrospinal Fluid and Blood-Brain Barriers Imaged by 18F-Labeled Metabolites of 18F-Setoperone Studied in Humans Using Positron Emission Tomography

18F-Setoperone, a sensitive radioligand for brain serotonin 5-HT2 receptor positron emission tomography studies, is metabolized into 18F-labeled metabolites, which participate in blood 18F radioactivity. Its main metabolite, identified as reduced 18F-setoperone, was synthesized and studied in humans to determine if 18F-labeled metabolites of 18F-setoperone (a) enter into the brain, (b) bind to the 5-HT2 receptor, and (c) explain the increase of 18F radioactivity in the free fraction in blood measured following 18F-setoperone injection. After reduced 18F-setoperone injection, the brain-to-blood 18F radioactivity concentration ratio (a) was low, at the beginning, indicating that this metabolite did not cross the blood-brain barrier; (b) was increased thereafter, with a higher radioactivity level in the choroid plexus than in brain tissue, suggesting a blood-CSF barrier crossing due to radioligand hydrophilicity; and (c) showed similar kinetics for cerebellum and frontal cortex, indicating that radioactive metabolites of 18F-setoperone did not bind to the 5-HT2 receptor. Because hydrophilic 18F-labeled metabolites of 18F-setoperone increased 18F radioactivity in the free fraction in blood, we quantified the relation between 18F-setoperone metabolism and free fraction kinetics in blood. A significant negative correlation was found between metabolism and free fraction rate constants in blood, showing it was possible to predict the 18F-setoperone metabolism rate using free fraction kinetics in blood. This will allow us to avoid the use of radio-TLC, a reference method that is difficult to use when multiple samples must be analyzed. A hydrophilic positron-emitter radioligand could also be used to study the blood-CSF barrier.     

Mapping of Histamine H1 Receptors in the Human Brain Using [11C]Pyrilamine and Positron Emission Tomography

We have studied the characteristics of carbon-11 labeled pyrilamine as a radioligand for investigating histamine H1 receptors in human brain with positron emission tomography (PET). [11C]Pyrilamine is distributed evenly in proportion to cerebral blood flow at initial PET images. Later (after 45-60 min), 11C radioactivity was observed at high concentrations in the frontal and temporal cortex, hippocampus, and thalamus, and at low concentrations in the cerebellum and pons. The regional distribution of the carbon-11 labeled compound in the brain corresponded well with that of the histamine H1 receptors determined in vitro in autopsied materials. In six controls, the frontal and temporal cortices/cerebellum ratio increased during the first 60 min to reach a value of 1.22 +/- 0.071. Intravenous administration of d-chlorpheniramine (5 mg) completely abolished the specific binding in vivo in the frontal cortex and temporal cortex (cortex/cerebellum ratio, 0.955 +/- 0.015). The availability of this method for measuring histamine H1 receptors in vivo in humans will facilitate studies on neurological and psychiatric disorders in which histamine H1 receptors are thought to be abnormal.