Metabolic hormones play essential regulatory roles in many biological processes, including morphogenesis, growth, and reproduction through the maintenance of energy balance. Various metabolic hormones originally discovered in mammals, including ghrelin, leptin, and <em>nesfatin</em>-<em>1</em> have been identified and characterized in fish. However, physiological roles of these metabolic hormones in regulating reproduction are largely unknown in fishes, especially in males. While the information available is restricted, this review attempts to summarize the main findings on the roles of metabolic peptides on the reproductive system in male fishes with an emphasis on testicular development and spermatogenesis. Specifically, the primary goal is to review the physiological interactions between hormones that regulate reproduction and hormones that regulate metabolism as a critical determinant of testicular function. A brief introduction to the localization of metabolic hormones in fish testis is also provided. Besides, the consequences of fasting and food deprivation on testicular development and sperm quality will be discussed with a focus on interactions between metabolic and reproductive hormones.

We examined whether the chemogenetic activation of endogenous arginine vasopressin (AVP) affects central <em>nesfatin</em>-<em>1</em>/NucB2 neurons, using a transgenic rat line that was previously generated. Saline (<em>1</em> mL/kg) or clozapine-N-oxide (CNO, <em>1</em> mg/mL/kg), an agonist for hM3Dq, was subcutaneously administered in adult male AVP-hM3Dq-mCherry transgenic rats (300-370 g). Food and water intake were significantly suppressed after subcutaneous (s.c.) injection of CNO, with aberrant circadian rhythmicity. The percentages of Fos expression in <em>nesfatin</em>-<em>1</em>/NucB2-immunoreactive neurons were significantly increased in the hypothalamus and brainstem at <em>1</em>20 min after s.c. injection of CNO. Suppressed food intake that was induced by chemogenetic activation of endogenous AVP was ablated after intracerebroventricularly administered <em>nesfatin</em>-<em>1</em>/NucB2-neutralizing antibody in comparison with vehicle, without any alteration of water intake nor circadian rhythmicity. These results suggest that chemogenetic activation of endogenous AVP affects, at least in part, central <em>nesfatin</em>-<em>1</em>/NucB2 neurons and may exert anorexigenic effects in the transgenic rats.

<strong class="sub-title"> Keywords: </strong> Arginine vasopressin; DREADDs; Feeding; Hypothalamus; Nesfatin-<em>1</em>/NucB2; Transgenic rat.

The lack of reliable plasma biomarkers limits their use in the diagnosis of gastric cancer (GC). The current study aimed to determine whether plasma <em>nesfatin</em>-<em>1</em> can be used as a novel non-invasive biomarker for the diagnosis of GC. The levels of <em>nesfatin</em>-<em>1</em> in 40 patients with GC and 40 healthy individuals, who were selected from the Chaohu Hospital Affiliated to Anhui Medical University, were assessed. ELISA was used for the measurement of plasma <em>nesfatin</em>-<em>1</em> levels, while immunohistochemistry was applied to determine Ki67 protein expression in GC and normal gastric tissues. The diagnostic value of plasma <em>nesfatin</em>-<em>1</em> for GC was further assessed using receiver operating characteristic (ROC) curve analysis. The results revealed that, compared with the controls, the mean <em>nesfatin</em>-<em>1</em> levels in patients with GC were significantly increased. Furthermore, the protein expression of Ki67 in GC tissue was significantly upregulated compared with that in normal gastric tissue. Plasma <em>nesfatin</em>-<em>1</em> levels were also demonstrated to be correlated with Ki67 protein expression in GC tissues. Additionally, ROC curve analysis indicated the potential diagnostic value of <em>nesfatin</em>-<em>1</em>, and the area under the ROC curve (AUC) for <em>nesfatin</em>-<em>1</em> was 0.857 (95% confidence interval, 0.769-0.946). At a threshold <em>nesfatin</em>-<em>1</em> level of <em>1</em>.075 ng/ml, the optimal sensitivity and specificity were 70.0 and 95.0%, respectively, in discriminating patients with GC from healthy controls. These results indicated that plasma <em>nesfatin</em>-<em>1</em> may serve as a novel biomarker for the diagnosis of GC and determination of GC cell proliferation.

<em>Nesfatin</em>-<em>1</em> is a novel anorexic peptide hormone that also exerts cardiovascular protective effects in rodent models. However, <em>nesfatin</em>-<em>1</em> treatment at high doses also exerts vasopressor effects, which potentially limits its therapeutic application. Here, we evaluated the vasoprotective and vasopressor effects of <em>nesfatin</em>-<em>1</em> at different doses in mouse models. Wild-type mice and those with the transgene nucleobindin-2, a precursor of <em>nesfatin</em>-<em>1</em>, were employed. Wild-type mice were randomly assigned to treatment with vehicle or <em>nesfatin</em>-<em>1</em> at 0.2, 2.0 or <em>1</em>0 μg/kg/day (Nes-0.2, Nes-2, Nes-<em>1</em>0, respectively). Subsequently, mice underwent femoral artery wire injury to induce arterial remodeling. After 4 weeks, injured arteries were collected for morphometric analysis. Compared with vehicle, <em>nesfatin</em>-<em>1</em> treatments at 2.0 and <em>1</em>0 μg/kg/day decreased body weights and elevated plasma <em>nesfatin</em>-<em>1</em> levels with no changes in systolic blood pressure. Furthermore, these treatments reduced neointimal hyperplasia without inducing undesirable remodeling in injured arteries. However, <em>nesfatin</em>-<em>1</em> treatment at 0.2 μg/kg/day was insufficient to elevate plasma <em>nesfatin</em>-<em>1</em> levels and showed no vascular effects. In nucleobindin-2-transgenic mice, blood pressure was slightly higher but neointimal area was lower than those observed in littermate controls. In cultured human vascular endothelial cells, <em>nesfatin</em>-<em>1</em> concentration-dependently increased nitric oxide production. Additionally, <em>nesfatin</em>-<em>1</em> increased AMP-activated protein kinase phosphorylation, which was abolished by inhibiting liver kinase B<em>1</em>. We thus demonstrated that <em>nesfatin</em>-<em>1</em> treatment at appropriate doses suppressed arterial remodeling without affecting blood pressure. Our findings indicate that <em>nesfatin</em>-<em>1</em> can be a therapeutic target for improved treatment of peripheral artery disease.

Feeding is a complex behaviour entailing elaborate interactions between forebrain, hypothalamic and brainstem neuronal circuits via multiple orexigenic and anorexigenic neuropeptides. Nucleobindin-2 (NUCB2)/<em>nesfatin</em>-<em>1</em> is a negative regulator of food intake and body weight with a widespread distribution in rodent brainstem nuclei. However, its localisation pattern in the human brainstem is unknown. The present study aimed to explore NUCB2/<em>nesfatin</em>-<em>1</em> immunoexpression in human brainstem nuclei and its possible correlation with body weight. Sections of human brainstem from 20 autopsy cases (<em>1</em>3 males, seven females; eight normal weight, six overweight, six obese) were examined using immunohistochemistry and double immunofluorescence labelling. Strong immunoreactivity for NUCB2/<em>nesfatin</em>-<em>1</em> was displayed in various brainstem areas, including the locus coeruleus, medial and lateral parabrachial nuclei, pontine nuclei, raphe nuclei, nucleus of the solitary tract, dorsal motor nucleus of vagus (<em>1</em>0N), area postrema, hypoglossal nucleus, reticular formation, inferior olive, cuneate nucleus, and spinal trigeminal nucleus. NUCB2/<em>nesfatin</em>-<em>1</em> was shown to extensively colocalise with neuropeptide Y and cocaine- and amphetamine-regulated transcript in the locus coeruleus, dorsal raphe nucleus and solitary tract. Interestingly, in the examined cases, NUCB2/<em>nesfatin</em>-<em>1</em> protein expression was lower in obese than normal weight subjects in the solitary tract (P = 0.020). The findings of the present study provide neuroanatomical support for a role for NUCB2/<em>nesfatin</em>-<em>1</em> in feeding behaviour and energy balance. The widespread distribution of NUCB2/<em>nesfatin</em>-<em>1</em> in the human brainstem nuclei may be indicative of its pleiotropic effects on autonomic, neuroendocrine and behavioural processes. In the solitary tract, a key integrator of energy status, altered neurochemistry may contribute to obesity. Further research is necessary to decipher human brainstem energy homeostasis circuitry, which, despite its importance, remains inadequately characterised.

<strong class="sub-title"> Keywords: </strong> brainstem; cocaine- and amphetamine-regulated transcript; <em>nesfatin</em>-<em>1</em>; neuropeptide Y; nucleobindin 2; nucleus of the solitary tract.

<em>Nesfatin</em>-<em>1</em> is the proteolytic cleavage product of Nucleobindin 2, which is expressed both in a number of brain nuclei (e. g., the paraventricular nucleus of the hypothalamus) and peripheral tissues. While Nucleobindin 2 acts as a calcium binding protein, <em>nesfatin</em>-<em>1</em> was shown to affect energy homeostasis upon central nervous administration by decreasing food intake and increasing thermogenesis. In turn, Nucleobindin 2 mRNA expression is downregulated in starvation and upregulated in the satiated state. Still, knowledge about the physiological role of endogenous Nucleobindin 2/<em>nesfatin</em>-<em>1</em> in the control of energy homeostasis is limited and since its receptor has not yet been identified, rendering pharmacological blockade impossible. To overcome this obstacle, we tested and successfully established an antibody-based experimental model to antagonize the action of <em>nesfatin</em>-<em>1</em>. This model was then employed to investigate the physiological role of endogenous Nucleobindin 2/<em>nesfatin</em>-<em>1</em>. To this end, we applied <em>nesfatin</em>-<em>1</em> antibody into the paraventricular nucleus of satiated rats to antagonize the presumably high endogenous Nucleobindin 2/<em>nesfatin</em>-<em>1</em> levels in this feeding condition. In these animals, <em>nesfatin</em>-<em>1</em> antibody administration led to a significant decrease in thermogenesis, demonstrating the important role of endogenous Nucleobindin 2/<em>nesfatin</em>-<em>1</em>in the regulation of energy expenditure. Additionally, food and water intake were significantly increased, confirming and complementing previous findings. Moreover, neuropeptide Y was identified as a major downstream target of endogenous Nucleobindin 2/<em>nesfatin</em>-<em>1</em>.

Studies of <em>nesfatin</em>-<em>1</em> in glucose metabolism have become a topic of interest recently, however, the specific receptor for <em>nesfatin</em>-<em>1</em> has not yet been identified. Some studies hinted at a connection between <em>nesfatin</em>-<em>1</em> and the ghrelin receptor, growth hormone secretagogue receptor. Therefore, we aimed to study the role of GHSR in the glycemic effects of <em>nesfatin</em>-<em>1</em> as well as its downstream pathways. We employed C57/BL6 mice (wild type and GHSR knockout mice) eating a normal chow diet and a high fat diet in this study, and the experimental technique included western blot, real-time PCR, immunofluorescence and ELISA. We found that in mice fed a normal chow diet (NCD), <em>nesfatin</em>-<em>1</em> improved glucose tolerance, up-regulated AKT kinase (AKT) mRNA levels and phosphorylation and GLUT4 membrane translocation in skeletal muscle. These effects were blocked by co-injection of GHSR antagonist [D-Lys3]-GHRP-6 and were attenuated in GHSR knockout mice. In mice fed high-fat diet (HFD), <em>nesfatin</em>-<em>1</em> not only exerted the effects observed in NCD mice, but also suppressed appetite and raised AKT levels in liver tissues that also required GHSR. Peripheral <em>nesfatin</em>-<em>1</em> suppressed c-fos expression of GHSR immunoreactive neurons induced by fasting in hypothalamic nuclei, indicating that <em>nesfatin</em>-<em>1</em> inhibited the activation of central GHSR. We concluded that the effects of <em>nesfatin</em>-<em>1</em> on food intake and glucose metabolism were GHSR-dependent, and that the glycemic effect was associated with AKT and GLUT4. This study should stimulate further exploration of the <em>nesfatin</em>-<em>1</em> receptor.

Nucleobindin2 (NUCB2)/<em>nesfatin</em>-<em>1</em> expression in human plasma positively correlates with the expression of pro-inflammatory cytokines in patients with chronic obstructive pulmonary disease (COPD), implicating its potential role in neutrophilic lung inflammation. There are no data on the localization of nucleobindin2 (NUCB2)/<em>nesfatin</em>-<em>1</em> in human lungs and inflammatory cells. We examined the localization of NUCB2/<em>nesfatin</em>-<em>1</em>-immunoreactivity in normal and inflamed human lungs obtained from COPD patients and neutrophils with light and immunoelectron microscopy. Immunohistology showed localization of NUCB2/<em>nesfatin</em>-<em>1</em>-like immunoreactivity in the bronchiolar epithelium, alveolar septa, vascular endothelium and various immune cells of normal and inflamed lungs. Further, NUCB2/<em>nesfatin</em>-<em>1</em>-like immunoreactivity accumulated within 0.5 μm of the plasma membrane in human neutrophils following 90 min. of <em>1</em> ng/mL LPS stimulation. NUCB2/<em>nesfatin</em>-<em>1</em>-like immunoreactivity was also found to localize in euchromatic portions of neutrophilic nuclei at five times the mean concentration compared to heterochromatin. Finally, our results indicate that NUCB2/<em>nesfatin</em>-<em>1</em>-like immunoreactivity is predominantly cytoplasmic including that in the Golgi complex and vesicles as it localizes at two times the concentration in neutrophilic cytoplasm compared to nucleus. Our study is the first to detail the localization of NUCB2/<em>nesfatin</em>-<em>1</em>-like immunoreactivity in lungs and neutrophils, and nuclear localization of NUCB2/<em>nesfatin</em>-<em>1</em> also implicates its potential role in transcriptional regulation.

<strong class="sub-title"> Keywords: </strong> Immuno-gold Electron Microscopy; Immunohistology; NUCB2; Nesfatin-<em>1</em>.

Neuropeptides are involved in various brain activities being able to control a wide spectrum of higher mental functions. The purpose of this concise structural investigation was to detect the possible immunoreactivity of the novel multifunctional neuropeptide <em>nesfatin</em>-<em>1</em> within the human bed nucleus of the stria terminalis (BNST). The BNST is involved in the mechanism of fear learning, integration of stress and reward circuits, and pathogenesis of addiction. <em>Nesfatin</em>-<em>1</em>-expressing neurons were identified for the first time in several regions of the BNST using both immunohistochemical and fluorescent methods. This may implicate a potential contribution of this neuropeptide to the BNST-related mechanisms of stress/reward responses in the human brain.

<em>Nesfatin</em>-<em>1</em> and neuronostatin in the central nervous system participate in regulating stress responses. Glucocorticoid hormones affect the brain through glucocorticoid receptors (GR). We investigated in the rat the possibility of co-localizing <em>nesfatin</em>-<em>1</em> and neuronostatin neurons in hypothalamic areas with GR. using immunohistochemistry. We counted <em>nesfatin</em>-<em>1</em> and neuronostatin stained neurons. We counted GR positive <em>nesfatin</em>-<em>1</em> neurons in the arcuate nucleus (ARC) and paraventricular nucleus (PVN) and GR positive neuronostatin neurons in the periventricular nucleus (PeN). The percentage of <em>nesfatin</em>-<em>1</em> neurons that expressed GR was 38.4% in female rats and 2<em>1</em>.9% in male rats in the ARC, and 33.3% in female rats and 29.2% in male rats in the PVN. The percentage of neuronostatin neurons that expressed GR was 39.<em>1</em>% in female rats and 39.9% in male rats in the PeN. We found that a substantial portion of <em>nesfatin</em>-<em>1</em> and neuronostatin neurons were stained for GR. We speculate that the pattern of GR might permit secretion of neuropeptides to be stimulated by peripheral glucocorticoid signals. Stress can suppress food intake, in part, through the GR in neurons that express <em>nesfatin</em>-<em>1</em>, which is a satiety molecule, and in neurons that express neuronostatin, which is an anorexigenic peptide.

<strong class="sub-title"> Keywords: </strong> Glucocorticoid; hypothalamus; <em>nesfatin</em>-<em>1</em>; neuronostatin; rat; receptors; stress.

The two peptides phoenixin and <em>nesfatin</em>-<em>1</em> are colocalized in hypothalamic nuclei involved in the mediation of food intake and behavior. Phoenixin stimulates food intake and is anxiolytic, while <em>nesfatin</em>-<em>1</em> is an anorexigenic peptide shown to increase anxiety and anhedonia. Interestingly, central activation of both peptides can be stimulated by restraint stress giving rise to a role in the mediation of stress. Thus, the aim of the study was to test whether also peripheral circulating levels of NUCB2/<em>nesfatin</em>-<em>1</em> and phoenixin are altered by restraint stress. Male ad libitum fed Sprague Dawley rats equipped with a chronic intravenous catheter were subjected to restraint stress and plasma levels of NUCB2/<em>nesfatin</em>-<em>1</em>, phoenixin and cortisol were measured over a period of 240 min and compared to levels of freely moving rats. Peripheral cortisol levels were significantly increased in restrained rats at 30, 60, <em>1</em>20 and 240 min compared to controls (p < 0.05). In contrast, restraint stress decreased plasma phoenixin levels at <em>1</em>5 min compared to unstressed conditions (0.8-fold, p < 0.05). Circulating NUCB2/<em>nesfatin</em>-<em>1</em> levels were increased only at 240 min in restrained rats compared to those in unstressed controls (<em>1</em>.3-fold, p < 0.05). In addition, circulating NUCB2/<em>nesfatin</em>-<em>1</em> levels correlated positively with phoenixin levels (r = 0.378, p < 0.00<em>1</em>), while neither phoenixin nor <em>nesfatin</em>-<em>1</em> were associated with cortisol levels (r = 0.0275, and r=-0.<em>1</em>43, p> 0.05). These data suggest that both peptides, NUCB2/<em>nesfatin</em>-<em>1</em> and phoenixin, are affected by restraint stress, although less pronounced than circulating cortisol.

<strong class="sub-title"> Keywords: </strong> Brain-gut axis; Cortisol; HPA axis; Nesfatin-<em>1</em>; Phoenixin; Restraint; Stress.

Diabetic retinopathy (DR) is a major complication of diabetes mellitus that may cause severe visual impairment. It has been reported that the levels of <em>nesfatin</em>-<em>1</em> in the serum and vitreous humor were negatively correlated with DR; however, its role in DR has not been fully elucidated. Therefore, the present study was performed to investigate the effect of <em>nesfatin</em>-<em>1</em> on high glucose-treated human retinal epithelial cells (ARPE-<em>1</em>9) and explore the underlying mechanism. The effects of <em>nesfatin</em>-<em>1</em> on cell viability, inflammation, oxidative stress and apoptosis were examined under high glucose conditions. The Cell Counting Kit-8 assay was used to determine cell viability. The levels of inflammatory cytokines were evaluated using ELISA kits. The reactive oxygen species and malondialdehyde content was estimated using commercial assay kits. Flow cytometry was performed to detect apoptotic cells and western blot analysis was employed to evaluate the expression of apoptosis-associated proteins. Moreover, the levels of NF-κB, NACHT, LRR and PYD domains-containing protein 3 (NLRP3) and high-mobility group protein B<em>1</em> (HMGB<em>1</em>) were determined via western blot analysis. The results revealed that <em>nesfatin</em>-<em>1</em> enhanced cell viability and suppressed inflammation, oxidative stress and apoptosis in the presence of high glucose concentration. Moreover, the activation of the NF-κB/NLRP3 inflammasome signaling and the expression of HMGB<em>1</em> were inhibited by <em>nesfatin</em>-<em>1</em>. Furthermore, HMGB<em>1</em> overexpression partially abrogated the inactivation of the NF-κB/NLRP3 inflammasome pathway caused by <em>nesfatin</em>-<em>1</em>. Taken together, these findings demonstrated that <em>nesfatin</em>-<em>1</em> inhibited the activation of the NF-κB/NLRP3 inflammasome signaling via modulating HMGB<em>1</em> and exerted a protective effect on ARPE-<em>1</em>9 cells against high glucose-induced inflammation, oxidative stress and apoptosis.

<strong class="sub-title"> Keywords: </strong> LRR and PYD domains-containing protein 3; NACHT; diabetic retinopathy; high glucose; high-mobility group protein B<em>1</em>; <em>nesfatin</em>-<em>1</em>.

<strong class="sub-title"> Background: </strong> Phoenixin, spexin and <em>nesfatin</em>-<em>1</em> belong to a family of newly discovered multifunctional neuropeptides that play regulatory roles in several brain structures and modulate the activity of important neural networks. However, little is known about their expression and action at the level of brainstem. The present work was, therefore, focused on gene expression of the aforementioned peptides in the brainstem of rats chronically treated with olanzapine, a second generation antipsychotic drug.

<strong class="sub-title"> Methods: </strong> Studies were carried out on adult, male Sprague-Dawley rats that were divided into 2 groups: control and experimental animals treated with olanzapine (28-day-long intraperitoneal injection, at dose 5 mg/kg daily). All individuals were killed under anesthesia and the brainstem excised. Total mRNA was isolated from homogenized samples of both structures and the RT-PCR method was used for estimation of related SMIM20/phoenixin, NPQ/spexin and NUCB2/<em>nesfatin</em>-<em>1</em> gene expression.

<strong class="sub-title"> Results: </strong> Long-term treatment with olanzapine is reflected in qualitatively different changes in expression of examined neuropeptides mRNA in the rat brainstem. Olanzapine significantly decreased NPQ/spexin mRNA expression, but increased SMIM20/phoenixin mRNA level in the rat brainstem; while NUCB2/<em>nesfatin</em>-<em>1</em> mRNA expression remained unchanged.

Conclusions: Olanzapine can affect novel peptidergic signaling in the rat brainstem. This may cautiously suggest the presence of an alternative mode of its action.

<strong class="sub-title"> Keywords: </strong> Brainstem; Nesfatin-<em>1</em>; Olanzapine; Phoenixin; Spexin.

Ghrelin and <em>nesfatin</em>-<em>1</em> are two recently discovered peptide hormones that play opposite roles in the food intake, body-weight control and energy homeostasis in both human and rodents. Beyond its appetite-control function, increasing evidence has shown that ghrelin affects multiple advanced activities in the central nervous system, including memory and emotion. <em>Nesfatin</em>-<em>1</em> was also widely expressed in extra-hypothalamic brain regions including hippocampus and amygdala. However, the possible actions of <em>nesfatin</em>-<em>1</em> in those important brain regions are largely unknown. In this study, we micro-infused ghrelin or <em>nesfatin</em>-<em>1</em> into the lateral amygdala (LA) or area CA<em>1</em> of the dorsal hippocampus (CA<em>1</em>) and investigated the immediate effects of those two peptide hormones on cognitive and affective behaviors. We found that the micro infusion of ghrelin into the LA or the CA<em>1</em> interfered with certain types of learning and memory in both rats and mice, while <em>nesfatin</em>-<em>1</em> had no effect. Our data thus suggested that although <em>nesfatin</em>-<em>1</em> works as a functional antagonist of ghrelin in the feeding control, only ghrelin affects learning and memory.

<b>Background:</b> Glioblastoma is one of the most malignant tumors in the brain with high mortality. In recent years, immunotherapy and targeted therapy show great prospects in the treatments for glioblastoma, whereas more effective therapeutic targets are still urgently needed to be developed. Nucleobindin-2 (NUCB2) is the precursor protein of <em>nesfatin</em>-<em>1</em>, which have a variety of metabolic functions, such as food intake and temperature regulation. In recent years, the potential link between NUCB2 and the development of multiple cancer was gradually revealed; however, the effects of NUCB2 on the progression of glioblastoma are still unclear. <b>Methods:</b> Immunohistochemical assays were performed to explore the NUCB2 expression levels in 94 samples of glioblastoma and corresponding nontumor tissues; patients were divided into NUCB2 high expression group and low expression group. Clinical analysis related to the clinical features, the potential link between NUCB2 expression levels, and clinical features were analyzed; the effects of NUCB2 on cell proliferation and invasion of glioblastoma were detected through colony formation and MTT assay, and transwell assay respectively. The possible effects of NUCB2 on tumor growth and metastasis were measured in mice. <b>Results:</b> In this study, we demonstrated that NUCB2 over-expression was correlated with the high degree of recurrence of patients with glioblastoma. Further, we also revealed that NUCB2 promoted cell proliferation and invasion of glioblastoma <i>in vitro</i> and promoted the growth and metastasis of glioblastoma in mice. <b>Conclusion:</b> This study provided evidence that NUCB2 might be a novel therapeutic target of glioblastoma.

In recent researches, high expression of <em>nesfatin</em>-<em>1</em>/nucleobindin-2 (NUCB2) is linked to poor prognosis in prostate and colon cancer due to the enhancement in proliferation, migration, and invasion. However, the role of <em>nesfatin</em>-<em>1</em> in bladder cancer is not clear. In this study, we examined the expression of NUCB2 in bladder cancer using immunohistochemistry and observed that its high expression was associated with recurrence and metastasis. In addition, the transwell assay and wound healing assay showed that cell migration and invasion were decreased with NUCB2 knockdown in T24 and 5637 cells. In vivo, tumor growth and metastasis were inhibited with shRNA treatment in T24 cells. Those results showed that NUCB2 played an important role in bladder cancer and could be considered a potent prognostic factor in bladder cancer.

OBJECTIVE

To determine whether Nucb2/<em>nesfatin</em><em>1</em> production is regulated by the cannabinoid system through the intracellular mTOR pathway in the stomach.

METHODS

Sprague Dawley rats were treated with vehicle, rimonabant, rapamycin or rapamycin+rimonabant. Gastric tissue obtained from the animals was used for biochemical assays: Nucb2 mRNA measurement by real time PCR, gastric Nucb2/nesfatin protein content by western blot, and gastric explants to obtain gastric secretomes. Nucb2/nesfatin levels were measured in gastric secretomes and plasma using enzyme-linked immunosorbent assay.

RESULTS

The inhibition of cannabinoid receptor <em>1</em> (CB<em>1</em>) by the peripheral injection of an inverse agonist, namely rimonabant, decreases food intake and increases the gastric secretion and circulating levels of Nucb2/<em>nesfatin</em>-<em>1</em>. In addition, rimonabant treatment activates mTOR pathway in the stomach as showed by the increase in pmTOR/mTOR expression in gastric tissue obtained from rimonabant treated animals. These effects were confirmed by the use of a CB<em>1</em> antagonist, AM28<em>1</em>. When the intracellular pathway mTOR/S6k was inactivated by chronic treatment with rapamycin, rimonabant treatment was no longer able to stimulate the gastric secretion of Nucb2/<em>nesfatin</em>-<em>1</em>.

CONCLUSIONS

The peripheral cannabinoid system regulates food intake through a mechanism that implies gastric production and release of Nucb2/Nesfatin-<em>1</em>, which is mediated by the mTOR/S6k pathway.

Fibroblast growth factor 2<em>1</em> (FGF2<em>1</em>) is a member of the FGF superfamily that acts in an endocrine manner. FGF2<em>1</em> is a key regulator of energy balance and metabolism in mammals, and has emerged as a therapeutic potential for treating obesity and diabetes. Here, we report that mRNAs encoding FGF2<em>1</em> and its receptors are widely distributed within the zebrafish tissues and are importantly modulated by fasting (decreased in brain and liver, and increased in gut). FGF2<em>1</em> stimulates food intake in zebrafish, likely in part by modulating brain npy/agrp and nucb2/<em>nesfatin</em>-<em>1</em> and gut ghrelin and cck mRNA expression. In accordance with this orexigenic role, the expression of FGF2<em>1</em> and its receptors were observed to increase preprandially and decrease post-feeding in the foregut and/or liver. Finally, we found important evidence in favor of a role for FGF2<em>1</em> in regulating glucose and lipid metabolism in the zebrafish liver in a way that mimics a fasting metabolic state.

The goal of this study was to determine whether the plasma leptin, <em>nesfatin</em>-<em>1</em>, cortisol, brain-derived neurotrophic factor (BDNF), and inflammatory cytokines could be used as potential biomarkers for the degree of craving in the alcohol-dependent patients after <em>1</em> month of abstinence. A total of 83 patients with alcohol use disorder (AUD) and 6<em>1</em> healthy subjects were assessed. Patients with AUD were selected from Department of Material Dependence, Anhui Mental Health Center, and subjects in the control group were selected from healthy volunteers. The Alcohol Urge questionnaire Scale (AUQ) was used to evaluate the extent of craving for alcohol, and the Michigan Alcoholism Screening Test (MAST), the Fagerstrom Test for Nicotine Dependence (FTND), the Self-Rating Anxiety Scale (SAS), and the Self-Rating Depression Scale (SDS) were also assessed in patients with AUD. Enzyme-Linked Immunosorbent Assay (ELISA) was used for the measurement of plasma leptin, <em>nesfatin</em>-<em>1</em>, cortisol, BDNF, Interleukin-6 (IL-6), C-reactive protein (CRP), and tumor necrosis factor-α (TNF-α) levels. Compare with healthy controls, the average leptin, leptin/BMI, IL-6, CRP, and TNF-α levels in patients with AUD were significantly increased, while the BDNF levels were significantly decreased. Moreover, the partial correlational analysis showed that the AUQ scores of the alcohol-dependent patients were positively correlated with the plasma leptin levels (<i>r</i> = 0.6<em>1</em>3, <i>P</i> < 0.00<em>1</em>), rather than <em>nesfatin</em>-<em>1</em> (<i>r</i> = 0.066, <i>P</i> = 0.569) after controlling for age as covariate. Furthermore, plasma <em>nesfatin</em>-<em>1</em> levels were found to be correlated with the SDS scores (<i>r</i> = 0.366, <i>P</i> = 0.00<em>1</em>) in the AUD group. In addition, plasma leptin levels were positively associated with the plasma IL-6 (<i>r</i> = 0.257, <i>P</i> = 0.033), CRP (<i>r</i> = 0.305, <i>P</i> = 0.0<em>1</em><em>1</em>), and TNF-α (<i>r</i> = 0.3<em>1</em><em>1</em>, <i>P</i> = 0.009) levels, and negatively associated with the BDNF levels (<i>r</i> = -0.245, <i>P</i> = 0.042) in patients with AUD. These results suggest that plasma leptin, but not <em>nesfatin</em>-<em>1</em>, might be a potential biomarker for the degree of craving in alcohol-dependent patients after <em>1</em> month of abstinence, the mechanism of which might be related to the dysfunction of the inflammatory cytokines and BDNF levels.

The regulation of food intake encompasses complex interplays between the gut and the brain. Among them, the gastrointestinal tract releases different peptides that communicate the metabolic state to specific nuclei in the hindbrain and the hypothalamus. The present overview gives emphasis on seven peptides that are produced by and secreted from specialized enteroendocrine cells along the gastrointestinal tract in relation with the nutritional status. These established modulators of feeding are ghrelin and <em>nesfatin</em>-<em>1</em> secreted from gastric X/A-like cells, cholecystokinin (CCK) secreted from duodenal I-cells, glucagon-like peptide <em>1</em> (GLP-<em>1</em>), oxyntomodulin, and peptide YY (PYY) secreted from intestinal L-cells and uroguanylin (UGN) released from enterochromaffin (EC) cells. © 202<em>1</em> American Physiological Society. Compr Physiol <em>1</em><em>1</em>:<em>1</em>679-<em>1</em>730, 202<em>1</em>.

<em>NESFATIN</em>-<em>1</em> acts as a neuroendocrine hormone to suppress gonadotropin secretion in the female goldfish and to prevent germinal vesicle breakdown of oocytes in the zebrafish. However, the expression and function of <em>NESFATIN</em>-<em>1</em> in meiotic maturation and development of porcine oocytes remains elusive. Genomic structure of porcine <em>NESFATIN</em>-<em>1</em> precursor nucleobindin 2 (NUCB2) is first characterized in detail and an evolutionally closer relationship of <em>NESFATIN</em>-<em>1</em> between pig and rat is shown by phylogenetic analysis of multiple species. Additionally, immunofluorescence analysis revealed that <em>NESFATIN</em>-<em>1</em> is predominantly expressed and localizes on the membrane of both theca cells and granulosa cells, but not expressed in oocytes. Real-time quantitative polymerase chain reaction showed that the abundance of <em>NESFATIN</em>-<em>1</em> transcripts in granulosa cells progressively decreases during the developmental transition from small follicles to large follicles. Correspondingly, <em>NESFATIN</em>-<em>1</em> could significantly enhance both the cleavage and blastocyst rate of parthenogenetically activated oocytes from small follicles (p < 0.05), whereas it did not affect meiotic maturation and development of oocytes from large follicles. Interestingly, we found that <em>NESFATIN</em>-<em>1</em> significantly improves meiotic maturation of oocytes cultured in chemically defined medium in the absence of pyruvate compared with the control group (p < 0.05), suggesting that the <em>NESFATIN</em>-<em>1</em> as a substitute for pyruvate exerts beneficial effects on porcine oocyte maturation. In conclusion, these results demonstrate that <em>NESFATIN</em>-<em>1</em> facilitates both meiotic maturation and development of porcine oocytes.

<em>Nesfatin</em>-<em>1</em> is a central anorectic peptide derived from the precursor protein nucleobindin-2 (NUCB2). In the present study, the changes in hypothalamic NUCB2 mRNA expression and their responses to food deprivation during the neonatal to pre-pubertal period (postnatal days <em>1</em>0, 20, and 30) were evaluated in male and female rats. The rats' serum leptin levels were also measured because NUCB2 mRNA expression is positively regulated by leptin. In both the female and male rats, hypothalamic NUCB2 mRNA expression tended to fall throughout development. Similarly, higher serum leptin levels were detected on postnatal day <em>1</em>0 than on postnatal days 20 and 30 in both sexes. Hypothalamic NUCB2 mRNA expression was positively correlated with the serum leptin level in both the female and male rats; however, the relationship was not significant in males. The hypothalamic NUCB2 mRNA levels of the fed and 24h fasted groups did not differ at any time point in either sex. On the other hand, the serum leptin levels of the 24h fasted group were significantly lower than those of the fed group at all time points in both sexes. It can be speculated that the upregulation of hypothalamic leptin activity might induce a transient increase in hypothalamic NUCB2 mRNA expression during the early postnatal period (postnatal day <em>1</em>0) in both sexes. However, hypothalamic NUCB2 mRNA expression does not become sensitive to a negative energy balance during the neonatal to pre-pubertal period.

<strong class="sub-title"> Objective: </strong> Delta-like <em>1</em> (DLK<em>1</em>) is known to inhibit adipocyte differentiation and <em>nesfatin</em>-<em>1</em> is a neuropeptide that plays a role in the regulation of nutrition and metabolism. We aimed to assess both the levels of DLK<em>1</em> and <em>nesfatin</em>-<em>1</em> in polycystic ovary syndrome (PCOS) and determine the association of DLK<em>1</em> and <em>nesfatin</em>-<em>1</em> with metabolic parameters.

<strong class="sub-title"> Materials and methods: </strong> Forty-four patients with PCOS and 40 healthy women as the control group were included in this study. Venous blood samples of the participants were collected, and hormonal, metabolic parameters, DLK<em>1</em> and <em>nesfatin</em>-<em>1</em> blood levels were determined. Anthropometric parameters were also determined. For a double comparison, the Mann-Whitney U test was used for non-parametric numerical data, and Student's t-test was used for parametric numerical data. Bivariate correlations were investigated using Spearman's correlation analysis. The diagnostic performance of the parameters was evaluated using receiver operating characteristic curve analysis.

<strong class="sub-title"> Results: </strong> The findings showed that DLK<em>1</em> and <em>nesfatin</em>-<em>1</em> levels were lower among the PCOS group, and the differences in these values were found to be statistically significant. A significant negative correlation was found between DLK<em>1</em> levels and body mass index (BMI), waist/hip ratio, visceral adiposity index (VAI), fasting serum insulin (FSI), homeostasis model of assessment-insulin resistance (HOMA-IR) and triglyceride levels. A significant negative correlation was found between <em>nesfatin</em>-<em>1</em> levels and BMI, VAI, FSI, HOMA-IR and triglyceride.

<strong class="sub-title"> Conclusion: </strong> The findings showed that DLK<em>1</em> and <em>nesfatin</em>-<em>1</em> levels were lower in PCOS. Based on this study, DLK<em>1</em> may be culpable for metabolic disorders in PCOS and can be a novel marker for PCOS in the future.

<strong class="sub-title"> Keywords: </strong> Polycystic ovary syndrome; delta-like <em>1</em>; insulin resistance; <em>nesfatin</em>-<em>1</em>; visceral adiposity index.

Polycystic ovary syndrome (PCOS) is a complex and heterogeneous endocrine disorder, generally exhibiting the characteristic features of hyperandrogenemia, insulin resistance (IR) and obesity. <em>Nesfatin</em>-<em>1</em> is derived from the precursor nucleobindin2 (NUCB2), and plays an active role in energy balance, glucose metabolism and most likely gonadal function. In order to explore the role of <em>nesfatin</em>-<em>1</em>, we employed a rat model that uses letrozole to induce PCOS. The PCOS rats exhibited increased body weight, irregular cycles, polycystic ovaries characterized by cysts formed from atretic follicles, and a diminished granulosa layer. The expression of both <em>nesfatin</em>-<em>1</em> mRNA and protein in the ovarian tissues of PCOS group decreased significantly compared to the control group (p < 0.05). <em>Nesfatin</em>-<em>1</em> expression in peripheral blood also decreased in the PCOS group, in contrast with the control group. Furthermore, we found that <em>nesfatin</em>-<em>1</em> had a positive correlation with FSH, E2 and P, whereas it had a negative correlation with LH, and total T (p < 0.05). When taken together, these data indicated that the decrease in <em>nesfatin</em>-<em>1</em> may contribute to the mechanism governing PCOS, and might provide a new potential target for therapies aimed at treating PCOS.