An injection of orexin A or B into the cisterna magna or the rostral ventrolateral medulla (RVLM), where bulbospinal vasomotor neurons are located, elevated arterial pressure (AP) and heart rate (HR). We examined how orexins affected RVLM neurons to regulate cardiovascular functions by using in vitro recordings of neuronal activity of the RVLM and in vivo measurement of cardiovascular functions in rats. Orexin A and B concentration-dependently depolarized RVLM neurons. At 100 nM, both peptides excited 42% of RVLM neurons. Tetrodotoxin failed to block orexin-induced depolarization. In the presence of N-(2-methyl-6-benzoxazolyl)-NЈ-1, 5-naphthyridin-4-yl urea (SB-334867), an orexin 1 receptor (OX 1 R) antagonist, orexin A depolarized 42% of RVLM neurons with a smaller, but not significantly different, amplitude (4.9 Ϯ 0.8 versus 7.2 Ϯ 1.1 mV). In the presence of (2S)-1-(3,4-dihydro-6,7-dimethoxy-2(1H)-isoquinolinyl)-3,3-dimethyl-2-[(4-pyridinylmethyl)amino]-1-butanone hydrochloride (TCS OX2 29), an orexin 2 receptor (OX 2 R) antagonist, orexin A depolarized 25% of RVLM neurons with a significantly smaller amplitude (1.7 Ϯ 0.5 mV). Coapplication of both antagonists completely eliminated orexin A-induced depolarization. An OX 2 R agonist, [Ala 11 ,D-Leu 15 ]-orexin B, concentration-dependently depolarized RVLM neurons. Regarding neuronal phenotypes, orexins depolarized 88% of adrenergic, 43% of nonadrenergic, and 36 to 41% of rhythmically firing RVLM neurons. Intracisternal TCS OX2 29 (3 and 10 nmol) suppressed intracisternal orexin A-induced increases of AP and HR, whereas intracisternal SB-334867 (3 and 10 nmol) had no effect on the orexin A-induced increase of HR but suppressed the orexin A-induced pressor response at 10 nmol. We concluded that orexins directly excite RVLM neurons, which include bulbospinal vasomotor neurons, and regulate cardiovascular function mainly via the OX 2 R, with a smaller contribution from the OX 1 R.Orexin A and orexin B (also known as hypocretin 1 and hypocretin 2) increase arterial pressure (AP), heart rate (HR), and sympathetic activity in rats and rabbits, when given by an intracerebroventricular (Samson et al., 1999;Shirasaka et al., 1999;Matsumura et al., 2001), intracisternal (Chen et al., 2000, or intrathecal (Antunes et al., 2001) injection. Regarding the sites of action of orexins in regulating cardiovascular and autonomic functions, the hypothalamic paraventricular nucleus was proposed to be one of the critical sites (reviewed in Kannan et al., 2007). In addition, the medullary structures essential for cardiovascular and autonomic regulation may be responsible for orexin-induced cardiovascular alterations. To explore the role of orexins in medullary control of the AP and HR, Chen et al. (2000) found that an intracisternal injection of orexins induced long-lasting pressor and positive chronotropic responses, and microinjection of orexin A into the rostral ventrolateral medulla (RVLM) caused similar responses in rats. The cardiovascular effects of orexins were also ex...

New Findings r What is the central question of this study?Central orexinergic activity is involved in tonic and phasic control of cardiovascular homeostasis. A potential role for elevated central orexinergic activity in the maintenance of hypertension in spontaneously hypertensive rats (SHRs) has not previously been explored. r What is the main finding and what is its importance?We show that central or intra-rostral ventrolateral medulla blockade of orexin 2 receptors produces a significant reduction of arterial pressure in SHRs, but not Wistar-Kyoto rats. This study demonstrates a previously unrecognized role of orexin 2 receptors in maintaining hypertension in SHRs.Orexins can raise arterial pressure and sympathetic activity and are involved in tonic and phasic control of cardiovascular homeostasis. We hypothesized that elevated central orexinergic activity contributes to the maintenance of hypertension in spontaneously hypertensive rats (SHRs). We examined this hypothesis by suppressing central orexinergic activity in SHRs and Wistar-Kyoto rats (WKYs) with specific antagonists or antibodies against orexin 1 (OX1R) and 2 receptors (OX2R). Intracerebroventricular administration of an OX1R antagonist, SB-334867 (30 and 100 nmol), induced no significant change in mean arterial pressure (MAP) and heart rate (HR) in SHRs and WKYs except that at 100 nmol it reduced HR in WKYs. In contrast, an OX2R antagonist, TCS-OX2-29 (3-30 nmol) induced long-lasting reductions of MAP and HR in SHRs (21 ± 3 mmHg and 22 ± 2 beats min −1 at 30 nmol), but not in WKYs. Intracerebroventricular anti-OX2R IgG, but not anti-OX1R IgG or non-immune goat IgG, significantly lowered MAP and HR in SHRs. None of the three IgGs affected MAP or HR in WKYs. The OX2R protein level in the rostral ventrolateral medulla (RVLM) was lower in SHRs than in WKYs, whereas no differences were found between SHRs and WKYs in the paraventricular hypothalamic nucleus, dorsomedialperifornical hypothalamic area or caudal nucleus tractus solitarii. The OX1R protein levels in these four regions did not differ between SHRs and WKYs. Injection of TCS-OX2-29 (50 pmol) into the RVLM produced a larger reduction of MAP in SHRs than in WKYs. We conclude that elevated OX2R-mediated activity in the brain, especially in the RVLM, may contribute to hypertension in SHRs.

What is the central question of this study? Our previous study demonstrates that elevated orexin 2 receptor (OX2R) activity within the rostral ventrolateral medulla (RVLM) contributes to hypertension in spontaneously hypertensive rats (SHRs), and a lower OX2R protein level was detected in their RVLM. The present study aims to explore the mechanisms underlying elevated orexinergic activity in the RVLM of SHRs, compared with their normotensive counterparts, Wistar-Kyoto rats. What is the main finding and its importance? Increased orexinergic input into the RVLM and enhanced OX2R responsiveness in the RVLM, which was mainly mediated by augmented OX2R-neuronal nitric oxide synthase signalling, may underlie the elevated OX2R activity within the RVLM of SHRs. Our previous study showed that elevated orexin 2 receptor (OX2R) activity within the rostral ventrolateral medulla (RVLM) contributes to hypertension in spontaneously hypertensive rats (SHRs). Herein, we investigated the mechanism(s) underlying the elevated OX2R activity. The following results were found. (i) More hypothalamic orexin A-immunoreactive (OXA-IR) cells existed in SHRs than in Wistar-Kyoto (WKY) rats at either 4 (2217 ± 43 versus 1809 ± 69) or 16 weeks of age (1829 ± 59 versus 1230 ± 84). The number of OXA-IR cells that project to the RVLM was higher in 16-week-old SHRs than in WKY rats (91 ± 11 versus 52 ± 11). (ii) Higher numbers of OXA-IR and RVLM-projecting OXA-IR cells were found in the dorsomedial and perifornical hypothalamus of 16-week-old SHRs. (iii) Spontaneously hypertensive rats had higher levels of orexin A and B in the hypothalamus and higher levels of orexin A in the RVLM than did WKY rats. (iv) Unilateral intra-RVLM application of OX2R agonist, orexin A or [Ala(11), d-Leu(15)]-orexin B (50 pmol) induced a larger pressor response in SHRs than in WKY rats. (v) Intra-RVLM pretreatment with a neuronal nitric oxide synthase (NOS) inhibitor, 7-nitro-indazole (2.5 pmol), or a soluble guanylate cyclase inhibitor, methylene blue (250 pmol), reduced the intra-RVLM [Ala(11), d-Leu(15) ]-orexin B-induced pressor response in both WKY rats and SHRs. In contrast, an inducible NOS inhibitor, aminoguanidine (100 pmol), was ineffective. (vi) Neuronal NOS was co-expressed with OX2R in RVLM neurons. In conclusion, increased orexinergic input and enhanced OX2R-neuronal NOS signalling may underlie elevated OX2R activity in the RVLM and contribute to the pathophysiology of hypertension in SHRs.

Internet addiction (IA) has become a global mental and social problem, which may lead to a series of psychiatric symptoms including uncontrolled use of internet, and lack of concentration. However, the exact pathophysiology of IA remains unclear. Most of functional connectivity studies were based on pre-selected regions of interest (ROI), which could not provide a comprehensive picture of the communication abnormalities in IA, and might lead to limited or bias observations. Using local functional connectivity density (lFCD), this study aimed to explore the whole-brain abnormalities of functional connectivity in IA. We evaluated the whole-brain lFCD resulting from resting-state fMRI data in 28 IA individuals and 30 demographically matched healthy control subjects (HCs). The correlations between clinical characteristics and aberrant lFCD were also assessed. Compared with HCs, subjects with IA exhibited heightened lFCD values in the right dorsolateral prefrontal cortex (DLPFC), left parahippocampal gyrus (PHG), and cerebellum, and the bilateral middle cingulate cortex (MCC) and superior temporal pole (STP), as well as decreased lFCD values in the right inferior parietal lobe (IPL), and bilateral calcarine and lingual gyrus. Voxel-based correlation analysis revealed the significant correlations between the Young's Internet Addiction Test (IAT) score and altered lFCD values in the left PHG and bilateral STP. These findings revealed the hyper-connectivity in cognitive control network and default mode network as well as the hypo-connectivity in visual attention network, verifying the common mechanism in IA and substance addiction, and the underlying association between IA, and attention deficit/hyperactivity disorder in terms of neurobiology.

A new monoterpenoid indole alkaloid compound (1) and six known monoterpenoid indole alkaloids compounds (2-7) were isolated from the barks of Voacanga africana Staph. The structures were established by spectral analysis as ibogamine-16-carboxylic acid,17,20-didehydro-5,6-dioxo-10-methoxy-methyl ester (1), voacamine (2), vobasine (3), voacangine (4), voacristine (5), 19-epi-voacristine (6) and 19-epi-heyneanine (7). Compound 1 was confirmed by X-ray crystallographic analysis. All of the isolated compounds were evaluated for cytotoxicity against five cell lines (HEPG-2, A375, MDA-MB-231, SH-SY5Y, CT26). Among them, compounds 2 and 6 displayed significant inhibitory activities, compounds 3, 4 and 5 showed moderate inhibitory activities, while compounds 1 and 7 showed no inhibitory activities against the five cell lines.

Purpose. Acupuncture is an effective therapy for Internet addiction (IA). However, the underlying mechanisms of acupuncture in relieving compulsive Internet use remain unknown. Neuroimaging studies have demonstrated the role of the ventral striatum (VS) in the progress of IA; hence, the aim of this study was to explore the effects of acupuncture on the resting-state functional connectivity (rsFC) and relevant network of VS in IA. Methods. Twenty-seven IA individuals and 30 demographically matched healthy control subjects (HCs) were recruited in this study. We acquired the functional magnetic resonance imaging (fMRI) data in IA subjects before and after 40 days of acupuncture treatment. Seed-to-voxel and ROI-to-ROI analyses were applied to detect the rsFC alterations of the VS and related network in IA subjects and to investigate the modulation effect of acupuncture on the rsFC. Results. Compared with HCs, IA subjects exhibited enhanced rsFC of the right ventral rostral putamen (VRP) with the left orbitofrontal cortex (OFC), premotor cortex (PMC), cerebellum, and right ventromedial prefrontal cortex (vmPFC). In the network including these five ROIs, IA also showed increased ROI-to-ROI rsFC. Using a paired t-test in IA subjects before and after 40 days of acupuncture, the increased ROI-to-ROI rsFC was decreased (normalized to HC) with acupuncture, including the rsFC of the right VRP with the left OFC, PMC, and cerebellum, and the rsFC of the left cerebellum with the left OFC, PMC, and right vmPFC. Furthermore, the change in rsFC strength between the right VRP and left cerebellum in IA individuals was found positively correlated with the Internet craving alleviation after acupuncture. Conclusions. These findings verified the modulation effect of acupuncture on functional connectivity of reward and habit systems related to the VS in IA individuals, which might partly represent the underlying mechanisms of acupuncture on IA.

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Although mounting neuroimaging studies have greatly improved our understanding of the neurobiological mechanism underlying internet addiction (IA), the results based on traditional group-level comparisons are insufficient in guiding individual clinical practice directly. Specific neuroimaging biomarkers are urgently needed for IA diagnosis and the evaluation of therapy efficacy. Therefore, this study aimed to develop support vector machine (SVM) models to identify IA and assess the efficacy of cognitive behavior therapy (CBT) based on unbiased functional connectivity density (FCD). Resting-state fMRI data were acquired from 27 individuals with IA before and after 8-week CBT sessions and 30 demographically matched healthy controls (HCs). The discriminative FCDs were computed as the features of the support vector classification (SVC) model to identify individuals with IA from HCs, and the changes in these discriminative FCDs after treatment were further used as features of the support vector regression (SVR) model to evaluate the efficacy of CBT. Based on the informative FCDs, our SVC model successfully differentiated individuals with IA from HCs with an accuracy of 82.5% and an area under the curve (AUC) of 0.91. Our SVR model successfully evaluated the efficacy of CBT using the FCD change ratio with a correlation efficient of 0.59. The brain regions contributing to IA classification and CBT efficacy assessment were the left inferior frontal cortex (IFC), middle frontal cortex (MFC) and angular gyrus (AG), the right premotor cortex (PMC) and middle cingulate cortex (MCC), and the bilateral cerebellum, orbitofrontal cortex (OFC) and superior frontal cortex (SFC). These findings confirmed the FCDs of hyperactive impulsive habit system, hypoactive reflecting system and sensitive interoceptive reward awareness system as potential neuroimaging biomarkers for IA, which might provide objective indexes for the diagnosis and efficacy evaluation of IA.