Combinatorial therapy with sub-effective Ro25-6981 and ZL006 ameliorates depressive-like behavior in single or combined stressed male mice.

Major depression is a severe neuropsychiatric disorder that poses a significant challenge to health. However, development of an effective therapy for the disease has long been difficult. Here, we investigate the efficacy of a novel combinatorial treatment employing sub-effective doses of Ro25-6981, an antagonist targeting GluN2B-containing NMDA receptors, in conjunction with ZL006, an inhibitor of the PSD95/nNOS, on mouse models of depression. We employed social isolation, chronic restraint stress, or a combination of both to establish a depressed mouse model. Treatment with the drug combination reduced depressive-like behaviors without affecting locomotor activity in mice subjected to social isolation or chronic restraint stress. Furthermore, the combination therapy ameliorated depressive-like behaviors induced by combined stress of chronic restraint followed by social isolation. Mechanistic studies revealed that the combined treatment downregulated the hippocampal nitric oxide level. However, the therapeutic benefits of this combination were negated by the activation of NMDA receptors with a low dose of NMDA or by increasing nitric oxide levels with l-arginine. Moreover, the combinatorial treatment had negligible effects on object memory and contextual fear memory. Our data establish a combined therapy paradigm, providing a potential strategy targeting major depression.

Disruption of Cdk5-GluN2B complex by a small interfering peptide attenuates social isolation-induced escalated intermale attack behavior and hippocampal oxidative stress in mice.

Social isolation has emerged as a significant issue during the COVID-19 pandemic that can adversely impact human mental health and potentially lead to pathological aggression. Given the lack of effective therapeutic interventions for aggressive behavior, alternative approaches are necessary. In this study, we utilized a genetic method combined with a pharmacological approach to identify and demonstrate the crucial role of Cdk5 in escalated intermale attack behavior induced by 2-week social isolation. Moreover, we developed a small peptide that effectively disrupts the interaction between Cdk5 and GluN2B, given the known involvement of this complex in various neuropsychiatric disorders. Administration of the peptide, either systemically or via intrahippocampal injection, significantly reduced oxidative stress in the hippocampus and attenuated intermale attack behavior induced by 2-week social isolation. These findings highlight the previously unknown role of the hippocampal Cdk5-GluN2B complex in social isolation-induced aggressive behavior in mice and propose the peptide as a promising therapeutic strategy for regulating attack behavior and oxidative stress.

The down-regulation of tyrosine phosphatase SHP2 activity is involved in the removal of surface AMPA receptors in long term depression.

Hebbian-type synaptic plasticity which includes long term potentiation (LTP) and long term depression (LTD), is the main cellular mechanism underlying learning and memory. Effective activity and synaptic content of tyrosine phosphatase SHP2 are required for AMPA receptor trafficking during LTP. However, the role of SHP2 in LTD has not been fully elucidated. This study shows that the phosphorylation level of SHP2 at Y542 decreased after LTD induction either in hippocampal cultures or acute CA1 mini slices. This change occurred at least 10 min after LTD induction and was alleviated by administration of NMDA receptor antagonist, APV. Furthermore, the SHP2 mutant (D61G), found in Noonan syndrome patients, prevented the removal of surface AMPA receptors during chemical-induced LTD on cultured hippocampal neurons. The results revealed a molecular basis of regulatory role of SHP2 in long term depression, thus expands our understanding of the SHP2 function in learning and memory.

Targeting GluN2B/NO Pathway Ameliorates Social Isolation-Induced Exacerbated Attack Behavior in Mice.

Exacerbated attack behavior has a profound socioeconomic impact and devastating social consequences; however, there is no satisfactory clinical management available for an escalated attack behavior. Social isolation (SI) is widespread during this pandemic and may exert detrimental effects on mental health, such as causing heightened attack behavior. To explore the therapeutic approaches that alleviate the SI-induced heightened attack behavior, we utilized pharmacological methods targeting the GluN2B/NO signaling pathway during the attack behavior. Ifenprodil and TAT-9C peptide targeting GluN2B showed that the inhibition of GluN2B mitigated the SI-induced escalated attack behavior and the SI-induced aberrant nitric oxide (NO) level in the brain. Additionally, the potentiation of the NO level by L-arginine reversed the effects of the inhibition of GluN2B. Moreover, we showed that high doses of L-NAME and 7-NI and subeffective doses of L-NAME in combination with ifenprodil or TAT-9C or subeffective doses of 7-NI plus ifenprodil or TAT-9C all decreased the SI-induced escalated attack behavior and reduced the NO level, further supporting the idea that GluN2B/NO signaling is a crucial modulator of the escalated attack behavior.

Disrupting phosphorylation of Tyr-1070 at GluN2B selectively produces resilience to depression-like behaviors.

Drugs targeting N-methyl-D-aspartate receptors (NMDARs) have been approved to treat major depressive disorder (MDD); however, the presence of undesirable psychotomimetic and cognitive side effects may limit their utility. In this study, we show that the phosphorylation levels of the GluN2B subunit at tyrosine (Y) 1070 increase in mice after both acute and chronic restraint stress (CRS) exposure. Preventing GluN2B-Y1070 phosphorylation via Y1070F mutation knockin produces effects similar to those of antidepressants but does not affect cognitive or anxiety-related behaviors in subject mice. Mechanistically, the Y1070F mutation selectively reduces non-synaptic NMDAR currents and increases the number of excitatory synapses in the layer 5 pyramidal neurons of medial prefrontal cortex (mPFC) but not in the hippocampus. Altogether, our study identifies phosphorylation levels of GluN2B-Y1070 in the mPFC as a dynamic, master switch guarding depressive behaviors, suggesting that disrupting the Y1070 phosphorylation of GluN2B subunit has the potential for developing new antidepressants.

Differential Roles of GluN2B in Two Types of Chemical-induced Long Term Potentiation-mediated Phosphorylation Regulation of GluA1 at Serine 845 in Hippocampal Slices.

Synaptic plasticity, such as long term potentiation (LTP) and long term depression (LTD), underlies the cellular mechanism of learning and memory. Chemical-induced LTP (cLTP), which facilitates biochemical analysis of molecular changes in brain slices or neuronal cultures, has been accepted as an in vitro model to explore synaptic plasticity. cLTP, by either forskolin and rolipram (F&R) or glycine, is thought to be dependent on NMDA receptor. However, subunit-specific dependence and regulation of the NMDA receptor in cLTP remain poorly understood. In the present study, we found that phosphorylation level of GluN2B at tyrosine 1472 was modulated by F&R-induced LTP but not by glycine-induced LTP in hippocampal slices. Furthermore, an increased phosphorylation level of GluA1 at serine 845 by F&R-induced LTP rather than glycine-induced LTP was dependent on the activation of GluN2B, which is supported by the results from GluN2B antagonists, small interfering peptide and CRISPR-Cas9-mediated knock out of GluN2B. Taken together, we reveal the significant role of GluN2B in F&R-induced LTP, uncovering the role of GluN2B subunit of NMDA receptor in a specified cLTP.

Antidiabetic Drug Metformin Ameliorates Depressive-Like Behavior in Mice with Chronic Restraint Stress via Activation of AMP-Activated Protein Kinase.

Depression is one of the most prevalent neuropsychiatric disorders in modern society. However, traditional drugs, such as monoaminergic agents, have defect showing lag response requiring several weeks to months. Additionally, these drugs have limited efficacy and high resistance rates in patients with depression. Thus, there is an urgent need to develop novel drugs or approaches for the treatment of depression. Here, using biochemical, pharmacological, genetic and behavioral methods, we demonstrate that metformin imparts a fast-acting antidepressant-like effect in naïve mice as well as stressed mice subjected to chronic restraint stress model. Moreover, inhibition of AMP-activated protein kinase (AMPK) activity by compound C or knock down of hippocampal AMPKα occluded the antidepressant-like effect induced by metformin. Our results suggest that metformin may be a viable therapeutic drug for the treatment of stress-induced depression via activation of AMPK.

Acute stress regulates phosphorylation of N-methyl-d-aspartate receptor GluN2B at S1284 in hippocampus.

Exposure to acute stress leads to diverse changes, which include either beneficial or deleterious effects on molecular levels that are implicated in stress-related disorders. N-methyl-d-aspartate receptor (NMDAR)-mediated signalings, are thought to be vital players in stress-related mental disorders as well as attractive therapeutic targets for clinical treatment. In the present study, we utilized acute stress models in mice to explore regulation of phosphorylation level of S1284 in GluN2B subunit of NMDAR. We found out that forced swimming and acute restraint stress increased phosphorylation level of S1284, while phosphorylation level of S1284 was unaltered after brief exposure to open field. Moreover, phosphorylation change of S1284 was negated by treatment of roscovitine which is believed to be a Cyclin-dependent kinase inhibitor. Besides, we showed well correlation of phosphorylation change of S1284 and immobility time during forced swimming. Collectively, our results demonstrated that phosphorylation level of S1284 in GluN2B was regulated by acute stress.

Phosphorylation of Tyrosine 1070 at the GluN2B Subunit Is Regulated by Synaptic Activity and Critical for Surface Expression of N-Methyl-D-aspartate (NMDA) Receptors.

The number and subunit composition of synaptic N-methyl-d-aspartate receptors (NMDARs) play critical roles in synaptic plasticity, learning, and memory and are implicated in neurological disorders. Tyrosine phosphorylation provides a powerful means of regulating NMDAR function, but the underling mechanism remains elusive. In this study we identified a tyrosine site on the GluN2B subunit, Tyr-1070, which was phosphorylated by a proto-oncogene tyrosine-protein (Fyn) kinase and critical for the surface expression of GluN2B-containing NMDARs. The phosphorylation of GluN2B at Tyr-1070 was required for binding of Fyn kinase to GluN2B, which up-regulated the phosphorylation of GluN2B at Tyr-1472. Moreover, our results revealed that the phosphorylation change of GluN2B at Tyr-1070 accompanied the Tyr-1472 phosphorylation and Fyn associated with GluN2B in synaptic plasticity induced by both chemical and contextual fear learning. Taken together, our findings provide a new mechanism for regulating the surface expression of NMDARs with implications for synaptic plasticity.

[Comparing extended vertical partial laryngectomy and cricohyoidoepiglottopexy in the treatment of laryngeal carcinoma].

OBJECTIVE: To compare the results of extended vertical partial laryngectomy (similar to modified supracricoid partial laryngectomy with cricohyoidoepiglottopexy) and cricohyoidoepiglottopexy in the treatment of laryngeal carcinoma. METHODS: Retrospectively analyzed on the results and prognosis in patients underwent extended vertical partial laryngectomy and cricohyoidoepiglottopexy between 1998 and 2005. The operation was similar to extended vertical partial laryngectomy. The healthy vocal cord and ventricular band as well as about 1/3 to 2/3 laminas of thyroid cartilage were removed. The healthy cricoarytenoid joint was reserved. The vocal cord, ventricular band, fixed or limitation of motion arytenoid cartilage and 2/3 laminas of thyroid cartilage in ill side were removed. The posteroinferior border of laminas of thyroid cartilage in both sides were reserved. The cricoid was lifted and fixed with hyoid epiglottis directly. Extended vertical partial laryngectomy group consisted of 37 patients with glottic carcinoma (stage T2 16 cases, stage T3 21 cases) and cricohyoidoepiglottopexy group consisted of 34 patients with glottic carcinoma (stage T2 12 cases, stage T3 21 cases, stage T4 1 case). RESULTS: Kaplan-Meier analysis was performed to calculate the survival rates. The three-year cumulative survival rate was 91.7% in extended vertical partial laryngectomy group and 87.5% in cricohyoidoepiglottopexy group respectively. There was no significant difference between the two groups (P > 0.05). The five-year cumulative survival rate was 80.6% in extended vertical partial laryngectomy group and 81.3% in cricohyoidoepiglottopexy group respectively. There was also no significant difference between the two groups (P > 0.05). The decannulation rate was 100% (37/37) in extended vertical partial laryngectomy group and 94.1% (32/34) in cricohyoidoepiglottopexy group respectively. The decannulation time was (14.0 + or - 2.3) days in extended vertical partial laryngectomy group and (19.0 + or - 4.6) days in cricohyoidoepiglottopexy group respectively. The incidence of aspiration was 2.7% (1/37) in modified group and 23.5 (8/34) in cricohyoidoepiglottopexy group respectively evaluated at 8th weeks post-operatively. The evaluation of deglutition disorder was analyzed by Ridit analysis in both groups and the results showed that there was significant difference between the two groups (U = 7.341, P < 0.001). The symptom of aspiration in extended vertical partial laryngectomy group was significant less than in cricohyoidoepiglottopexy group. CONCLUSIONS: Although the survival rate was not different between the two groups. The preservation of laryngeal function in extended vertical partial laryngectomy group was significant better than in cricohyoidoepiglottopexy group and extended vertical partial laryngectomy.

[Identification of differentially expressed genes in recurrent nasopharyngeal carcinoma and analysis of their chromosomal location].

OBJECTIVE: To identify differentially expressed genes in recurrent nasopharyngeal carcinoma (rNPC) by DNA microarrays, and analyze chromosomal localizations and molecular function by bioinformatics. METHODS: The primary nasopharyngeal carcinoma (pNPC) tissue samples and rNPC tissue samples were selected, and Affymetrix Gene1.0 ST gene chips were used to identify differential expressed genes in rNPC, and the bioinformatics was used to analyze their chromosomal localizations as well as molecular functions. RESULTS: A total of 44 genes were identified to be differential expressed in rNPC. Thirty-six genes were down regulated, 8 genes were up regulated. Functional classification of down-regulation genes showed that most genes (10 genes, 27.8%) belonged to the enzyme activity genes, followed by calcium ion binding genes (7 genes, 19.4%), protein binding genes (5 genes, 13.9%), receptor activity genes (4 genes, 11.1%), ATP binding genes (2 genes, 5.6%), transcription factor genes (2 genes, 5.6%), extracellular matrix binding and growth factor binding have 1 gene respectively (each accounted for 2.8%). In addition, the functions of 4 genes (11.1%) were unknown. Functional classification of up-regulation genes showed most genes (3 genes, 37.5%) were unknown, followed enzyme activity genes (2 genes, 25.0%), receptor activity, calcium ion binding and voltage-gated ion channel activity genes have 1 genes respectively (each accounted for 12.5%). These genes were localized randomly on the most the chromosomes, with a majority of them localized on chromosomes 1, 17. Chromosome 1 contained the most differentially expressed genes (10, 22.7%), followed by chromosomes 17 (5, 11.3%). CONCLUSIONS: The differential expressed genes in rNPC were supposed to be randomly distributed on most chromosomes, but the majorities were found on chromosomes 1, 17. Abnormality in three groups of genes, including in enzyme activity, calcium ion binding and protein binding associate genes, might play important roles in rNPC. Those genes need to be further studied.

[Quality of life, personality, coping style in the patients with advanced laryngeal cancer].

OBJECTIVE: To evaluate the relationship between the life quality of advanced laryngeal squamous cell cancer patients and their personality, coping style and other psychological factors. METHODS: The life quality were measured for 2 sub-groups of advanced laryngeal cancer patients and the normal control group. The Eysenck personality questionnaire (EPQ), coping style questionnaires (CSQ) and University of Washington-quality of life (UW-QOL) were used for life quality evaluation. The 2 sub-groups of the patients on the worse speech, job and ability (group I), the better speech, job and ability (group II), and normal control. RESULTS: (1) UW-QOL score: the total, activity, recreation, job and speech scores of group II were significantly higher than those of group I (P < 0.01). Group II was better than group I in appearance (P < 0.05). (2) EPQ score: the P and N scores in group II were lower than that in group I (P < 0.05). The E scores in group II were significantly higher than that in group I (P < 0.01). (3) CSQ score: the problem-saving factor and help-seeking factor in group II were more significantly lower than that in group I (P < 0.01). The self-blaming factor in group II were higher than that in group I (P < 0.05). (4) There was positive correlation between total scores of QOL and the problem-saving factor, help-seeking factor of CSQ, the E scores of EPQ (P < 0.05), there was negative correlation between total scores of QOL and the P scores, the N scores of EPQ (P < 0.05). CONCLUSIONS: The advanced laryngeal cancer patients of group II have better life quality, and their personalities showed more extroversive, stable feeling, adaptable, mature coping styles. The above characteristics may have good effects on the prognosis of advanced laryngeal cancer.