LncRNA NR2F1-AS1 Inhibits the Malignant Properties of Cervical Cancer Cells via Targeting miR-642a-3p/NR2F1 Axis

ABSTRACT Background: Cervical cancer (CC), as a serious menace to the health of women, has long been one of the most lethal gynecologic neoplasms throughout the world. Long non-coding RNA (LncRNA) NR2F1-AS1 has been documented to exert crucial functions in many malignant tumors. Nonetheless, the function and molecular mechanism of NR2F1-AS1 in CC remain completely unknown. Objective: This study aimed to explore the function and molecular mechanism of NR2F1-AS1 in CC. Methods: The expression levels of NR2F1-AS1, miR-642a-3p, NR2F1 in CC tissues, and cell lines were examined by reverse transcription real-time quantitative polymerase chain reaction. Cell viability, proliferation, migration, and invasion were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, colony formation and Transwell assays. The protein levels of epithelial-mesenchymal transition markers and NR2F1 in CC cells were assessed by Western blot analysis. The correlations among NR2F1-AS1, miR-642a-3p, and NR2F1 were estimated through luciferase reporter and RNA immunoprecipitation assays. Results: NR2F1-AS1 expression was clearly downregulated in CC tissues and cell lines. Molecular mechanistic experiments showed that NR2F1-AS1 overexpression upregulated NR2F1 expression in CC cells by directly binding to miR-642a-3p, and inhibiting by this way cell viability, proliferation, migration, and invasion in CC. Rescue assays showed that NR2F1 knockdown or miR-642a-3p overexpression offset NR2F1-AS1 upregulation-induced inhibition on CC cell malignant phenotypes. Conclusion: These findings revealed that NR2F1-AS1 played a tumor suppressor role in CC by mediating the miR-642a-3p/NR2F1 axis.

Novel NR2F1 variant identified by whole-exome sequencing in a patient with Bosch–Boonstra–Schaaf optic atrophy syndrome

Bosch–Boonstra–Schaaf optic atrophy syndrome (BBSOAS) is an extremely rare autosomal dominant disorder characterized by intellectual disability, developmental delay, seizures, hypotonia, hearing loss, and optic nerve atrophy. This syndrome is caused by loss-of-function variants in the nuclear receptor subfamily 2 group F member 1 ( NR2F1 ) gene. To date, approximately 80 patients have been reported with BBSOAS. Here, we describe a 3-year-old infant with delayed development, intellectual disability, strabismus, nystagmus, and optic atrophy with well-characterized features associated with BBSOAS. Whole-exome sequencing revealed a novel heterozygous missense mutation (NM_005654.6:c.437G>A, p.Cys146Tyr) in the NR2F1 gene. This missense variant is predicted to be deleterious by the protein prediction tools (SIFT, PolyPhen-2, and MutationTaster). To the best of our knowledge, this is the first patient with BBSOAS reported from Turkey.

Clinical and genetic characteristics of a Han Chinese family with autosomal recessive enhanced S-cone syndrome / 中华实验眼科杂志

Objective:To analyze the clinical phenotypes and pathogenic gene of a Han Chinese family with enhanced S-cone syndrome (ESCS).Methods:The method of pedigree investigation was adopted.A suspected ESCS Han Chinese family including 8 members of 3 generations was recruited in Henan Eye Hospital from June to September 2021.There was one patient in the family.A thorough ophthalmic examination of the proband was carried out to evaluate the phenotypes, including visual acuity, degree of strabismus, anterior segment and fundus, autofluorescence imaging, fluorescein fundus angiography, full-field electroretinogram (ERG), multifocal ERG, optical coherence tomography.DNA was extracted from peripheral blood samples from the proband and family members.The pathogenic gene and variation were screened by whole exome sequencing (WES).The variation and co-segregation were verified by Sanger sequencing.The deleteriousness of the variation was analyzed by SIFT, Polyphen2 and MutationTaster.The pathogenicity of the variation was evaluated in accordance with the American College of Medical Genetics and Genomics (ACMG) Standards and Guidelines.The analysis of amino acid sequence conservation was performed by SIFT.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Henan Eye Hospital (No.HNEECKY-2017[6]).Written informed consent was obtained from each subject.Results:This pedigree was consistent with autosomal recessive inheritance.The proband had clinical features such as night blindness, hyperopia, accommodative esotropia, peripheral retinal pigmentation, retinoschisis, and photopic ERG responses dominated by large-amplitude waves.Variations including a compound heterozygous variation, c.671C>T: p.S224L on exon 5 and c. 955G>A: p.E319K on exon 6 of NR2E3 were identified by WES.The variations were confirmed to be consistent with co-segregation.The both loci were missense variations, the variation frequency of which was 0 in the East Asian population via the gnomAD database.The variations were predicted to be deleterious by SIFT, Polyphen2 and MutationTaster.The c.671C>T variation was recorded with unknown significance in ClinVar database, and the c.955G>A variation was an unreported new locus.According to the ACMG Standards and Guidelines, the both variations were labeled as with uncertain clinical significance, and the corresponding amino acid sequences were highly conservative across multiple species. Conclusions:This family has the clinical characteristics of ESCS and meets the genetic diagnosis criteria.Two novel variations in NR2E3 gene, c.671C>T: p.S224L and 955G>A: p.E319K, are found.

Molecular Mechanism of Wendantang Against Schizophrenia in Rats： An Exploration Based on miRNA-219， NR2B， DISC1 and CaMKⅡγ / 中国实验方剂学杂志

ObjectiveTo study the effects of Wendantang on the expression of miRNA-219， N-methyl-D-aspartate receptor 2B （NR2B）， disrupted in schizophrenia 1 （DISC1）， and Ca2+/calmodulin-dependent protein kinase Ⅱγ （CaMKⅡγ） in the frontal lobe of rats with schizophrenia. MethodSixty rats were randomly divided into six groups， namely normal group， model group， high-， medium-， and low-dose Wendantang groups， and clozapine group， with 10 rats in each group. Rats in high-， medium-， and low-dose Wendantang groups were intragastric with 40， 20， and 10 g·kg-1 Wendantang， and the ones in clozapine group were intragastric with 0.02 g·kg-1 clozapine， those in normal and model group were intragastric with equal volume of normal saline， once a day. After 21 days of administration， rats in all groups except for the normal group were injected with 0.6 mg·kg-1 dizocilpine maleate （MK-801） into the left abdominal cavity for inducing acute schizophrenia. The stereotypic behavior and ataxia in rats were scored according to SAMS and HOFFMAN criteria. The morphological changes in the prefrontal cortex were observed by hematoxylin-eosin （HE） staining. The protein expression levels of NR2B, DISC1 and CaMKⅡγ in the frontal lobe was detected by Western blot. The mRNA expression levels of miRNA-219 was detected by real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. ResultCompared with normal group， the model group exhibited significantly increased stereotypic behavior and ataxia scores （P<0.01）， karyopyknosis and karyolysis in most neurons of the prefrontal cortex， and down-regulated NR2B， DISC1， and CaMKⅡγ protein expression （P<0.01） and miRNA-219， NR2B， DISC1， and CaMKⅡγ mRNA expression （P<0.01）. Compared with model group， Wendantang high-， medium-， and low-doses group lowered the scores of stereotypic behavior and ataxia at 50， 60 mmin（P<0.05，P<0.01）. In high- and medium-dose Wendantang groups， the neurons in the prefrontal cortex were densely arranged. The karyopyknosis and karyolysis were alleviated to varying degrees. The NR2B protein expression in the frontal lobe was up-regulated （P<0.01）. In the medium- and low-dose Wendantang groups， the DISC1 protein expression in the frontal lobe was up-regulated （P<0.05，P<0.01）. Wendantang at each dose significantly increased the CaMKⅡγ protein expression （P<0.05） and miRNA-219， NR2B， DISC1， and CaMKⅡγ mRNA expression in the frontal lobe （P<0.05，P<0.01）. ConclusionWendantang improves the scores of stereotypical behavior and ataxia， relieves the karyopyknosis and karyolysis of neurons in the prefrontal cortex， and increases the expression levels of miRNA-219， NR2B， DISC1， and CaMKⅡγ of rats with schizophrenia， so as to alleviate the schizophrenic-like symptoms and schizophrenia.

KIF17 Modulates Epileptic Seizures and Membrane Expression of the NMDA Receptor Subunit NR2B / 神经科学通报·英文版

Epilepsy is a common and severe brain disease affecting >65 million people worldwide. Recent studies have shown that kinesin superfamily motor protein 17 (KIF17) is expressed in neurons and is involved in regulating the dendrite-targeted transport of N-methyl-D-aspartate receptor subtype 2B (NR2B). However, the effect of KIF17 on epileptic seizures remains to be explored. We found that KIF17 was mainly expressed in neurons and that its expression was increased in epileptic brain tissue. In the kainic acid (KA)-induced epilepsy mouse model, KIF17 overexpression increased the severity of epileptic activity, whereas KIF17 knockdown had the opposite effect. In electrophysiological tests, KIF17 regulated excitatory synaptic transmission, potentially due to KIF17-mediated NR2B membrane expression. In addition, this report provides the first demonstration that KIF17 is modified by SUMOylation (SUMO, small ubiquitin-like modifier), which plays a vital role in the stabilization and maintenance of KIF17 in epilepsy.

The role of nuclear receptor transcription factor NR2F6 in tumor / 生物工程学报

Nuclear receptor subfamily 2, group F, member 6 (NR2F6) is a member of orphan nuclear receptors, which is expressed in major tissues and organs of the human body, and plays an important role in the regulation of various biological functions and gene expressions. Recent studies have shown that the expression of NR2F6 was up-regulated in a variety of malignant tumors and showed significant correlations with cancer progression. These findings triggered the widespread interest in understanding the relationship between NR2F6 and cancer development and progression. In addition, the latest studies have underscored that NR2F6 was involved in enhancing antitumor immune responses that could serve as a potential target for immune regulation. This review summarizes the biological functions of NR2F6 and its role in tumors, with the aim to provide new insights into effective cancer therapies.

Boonsta-Bosch-Schaff optic atrophy syndrome with infantile spasm: A case and literatures review / 解放军医学杂志

Objective To report a case of Boonsta-Bosch-Schaff optic atrophy syndrome (BBSOAS) with infantile spasm, its clinical features as well as diagnosis and treatment process, and review the relevant literature. Methods Retrospectively analyze the clinical data of a case of BBSOAS with infant spasm patient in the First Medical Center of PLA General Hospital, retrieve the databanks of online human Mendelian genetic database (OMIM), PubMed, CNKI and Wanfang Medical Online, explore the clinical characteristic of BBSOAS with infant spasm patients, the relationship between the phenotype - genotype, and the therapeutic effect. Results The patient was a 9-month-old boy admitted to hospital due to "intermittent convulsions for 4 months". The growth and development of the child delayed, gaze following was poor; fundus examination showed pale optic disc (atrophy, small optic disc), spasm attack, and electroencephalogram indicated hypsarrhythmia. Genetic test found de novo missense mutation in NR2F1 gene c.383G>A (p.YS128tyr), so diagnosed as BBSOAS and Infantile spasms. The spasticity was not controlled and hypsarrhythmia was still existed in electroencephalogram after adrerrmrticotropic hormone (ACTH) and a variety of antiepileptic drugs were administered. Fever occurred 2 weeks after out of hospital oral perampanel, spasms was completely controlled after the febrile retrograde. A total of 9 English references were obtained by searching multiple databases and manual screening, and a total of 46 cases of BBSOAS were found, among which 9 cases were complicated with infantile spasms. All the amino acid changes caused by NR2F1 gene mutation were located in DNA-binding domain, and optic nerve atrophy or/and hypoplasia were observed. Conclusions The new missense mutation of NR2F1 gene c.383G>A that caused BBSOAS and infantile spasm is no literature report before. In case of optic atrophy or hypoplasia associated with spasm in infants and young children, the doctor should consider the possibility of BBSOAS, and do the genetic examination if necessary. Perampanel may be a potential drug for spasm control in such children.

Transcranial direct current stimulation promotes hippocampal neurogenesis in mice with cerebral ischemia involving up-regulation of nmda receptors / 中国药理学通报

Aim To investigate the improving effect of transcranial direct current stimulation (tDCS) on endogenous hippocampal neurogenesis in mice with cerebral ischemiaand the possible mechanism. Methods The model of acute cerebral ischemia in mice was established by bilateral common carotid artery occlision (BCCAO). The pathological changes of mice were detected by hippocampal HE staining. The learning and memory function of mice was assessed by Morris water maze. The number of BrdU, DCX and BrdU/NeuN-positive cells was observed through immunofluorescence staining for detecting hippocampal neurogenesis. The mRNA and protein expressions of NMDAR subunits NR2a and NR2b in hippocampus were detected by qRT-PCR and Western blot. Results The neuronal damage in the hippocampal CA1 region was marked (P <0. 01), and the learning and memory function significantly decreased (P<0. 01) in cerebral ischemia mice, suggesting the successful establishment of cerebral ischemia model. At the same time, the number of BrdU, DCX and BrdU/NeuN positive cells was up-reg-ulated significantly (P < 0. 01 ) , indicating the occurrence of neurogenesis in hippocampus after cefebral ischemia. Treatment with tDCS significantly ameliorated the pathological damage in CA1 region of mice, improved learning and memory, and promoted hippocam-pal neurogenesis. Meanwhile, the mRNA and protein expression levels of NR2a and NR2b in hippocampus were also up-regulated (P < 0. 05 or P < 0.01). Conclusions tDCS can promote hippocampal neurogenesis and improve learning and memory function in cerebral ischemia mice, which may be related to theup-regula-tion ofNR2a and NR2b expression.

Role of NR2B and CREB in ginsenoside Rbl regulation of methamphetamine-induced conditioned place preference in rats / 中国药理学通报

Aim To study the effect of ginsenoside Rbl on methamphetamine-induced CPP in rats and to explore the role of NR2B/CREB in it. Methods METH(2mg·kg-1, i.p) was administered to establish METH-induced CPP model in rats. 0 ∼3 d was the adaptation stage and 4 ∼ 13 d was the experimental stage. METH (2 mg · kg-1, i. p) or saline (10 mg · kg-1, i. p) was injected every other day. Rb1 (10 mg · kg-1, i.p) or saline was pre-injected lh before injection of METH or saline. After perfusion, the hippocampus was isolated from brain on ice, and the expression levels of NR2B, CREB and p-CREB were detected by Western blot. Results The animal model of METH-induced CPP was successfully established. The rats were pretreated with Rbl (10 mg · kg-1) for 1 h, and the time that the rats stayed in drug-paired was significantly reduced compared with METH group. Western blot results showed that NR2B, p-CREB and p-CREB/CREB significantly increased in METH group and without altering CREB expression levels compared with control group. However, after pre-treated with Rbl, the expression levels of NR2B, p-CREB and p-CREB/CREB decreased compared with METH group. Conclusions METH can significantly induce CPP in rats. Rbl may inhibit METH-induced CPP in rats by regulating NR2B and p-CREB.

A Review on Chronic Pain in Rheumatoid Arthritis: A Focus on Activation of NR2B Subunit of N-Methyl-D-Aspartate Receptors

@#Chronic pain is a debilitating condition that occurs after tissue damage, which substantially affects the patient’s emotional state and physical activity. The chronic pain in rheumatoid arthritis (RA) is the result of various autoimmune-induced inflammatory reactions in the joints. Both types of peripheral and central pain processing can lead to sensitisation. Non-steroidal anti-inflammatory drugs (NSAIDs) and disease-modifying anti-rheumatic drugs (DMARDs) can result in potent anti-inflammatory effect. However, these drugs are not able to suppress the pain from RA for a prolonged period. For years, researchers have examined the role of the N-methyl-D-aspartic acid receptor 2B (NR2B) subunit of N-methyl-D-aspartate receptors (NMDAR) in chronic and neuropathic pain models. This NMDAR subtype can be found in at the peripheral and central nervous system and it represents an effective therapy for RA pain management. This review focuses on the NR2B subunit of NMDAR and the different pathways leading to its activation. Furthermore, specific attention is given to the possible involvement of NR2B subunit in the peripheral and central pathogenesis of RA.

Effect of Lactobacillus casei on the depressive behavior and the expression of NR1 and NR2A receptors in hippocampus of rats with postpartum depression induced by maternal separation / 中华行为医学与脑科学杂志

Objective@#To investigate the effect of Lactobacillus casei on depressive behavior and the expression of NR1, NR2A receptor in postpartum depression rats induced by maternal separation.@*Methods@#Twenty-four Sprague-Dawley female rats were randomly divided into control group(CON group) (n=8), postpartum depression group(PPD group) (n=8), postpartum depression with lactobacillus casei group(PPD+ Lactobacillus casei group) (n=8). Rats of the control group were given no interventions.Rats in PPD group and PPD+ Lactobacillus casei group were given maternal separation to establish depression model.And then the rats in PPD+ Lactobacillus casei group were treated with Lactobacillus casei(8×108CFU/(kg·d)) for 4 weeks after 14 days of maternal separation.The forced swimming test (FST) was employed to detect the depressive behaviors.Lactobacillus, Bifidobacterium, Enterococcus faecalis and Escherichia coli in cecum of rats and the expression of NR1 and NR2A mRNA in hippocampus of rats were detected by real-time fluorescence quantitative PCR.@*Results@#Compared with CON group ((157.50±8.13) s), the immobility time of PPD group((200.00±10.35) s) was significantly longer (t=-3.23, P<0.05). Compared with PPD group, the immobility time of PPD+ Lactobacillus casei group ((153.25±7.41) s) was significantly shortened (t=3.67, P<0.05), and the depressive behavior was improved.Compared with CON group, the mRNA expression of Lactobacillus ((1.47±0.08), t=-5.87, P<0.01), Enterococcus faecalis ((1.23±0.08), t=-2.85, P<0.05) and Escherichia coli( (1.33±0.07), t=-4.96, P<0.01) in caecum were significantly increased in PPD group, while that of Bifidobacterium decreased significantly((0.65±0.07), t=5.18, P<0.01). Compared with PPD group, the mRNA expression of Lactobacillus( (1.05±0.05), t=3.67, P<0.01), Enterococcus faecalis ((0.97±0.05), t=2.74, P<0.05) and Escherichia coli ((1.06±0.05), t=3.31, P<0.01) were significantly decreased in PPD+ Lactobacillus casei group, while that of Bifidobacterium increased significantly ((0.98±0.04), t=-4.26, P<0.01). Compared with CON group, the mRNA expression of NR1 receptor ((0.57±0.04), t=9.65, P<0.01) and NR2A receptor ((0.60±0.05), t=8.64, P<0.05) in hippocampus of rats in PPD group were significantly decreased.Compared with PPD group, the expression of NR1 receptor ((1.01±0.05), t=-5.39, P<0.01) and NR2A receptor ((0.98±0.07), t=-3.91, P<0.05) in hippocampus were increased significantly in PPD+ Lactobacillus casei group.@*Conclusion@#Lactobacillus casei can improve the depressive behavior of postpartum depression in rats, and improve the intestinal flora, which affect the expression of NR1 and NR2A in hippocampus.

Effect of Lactobacillus casei on the depressive behavior and the expression of NR1 and NR2A recep-tors in hippocampus of rats with postpartum depression induced by maternal separation / 中华行为医学与脑科学杂志

Objective To investigate the effect of Lactobacillus casei on depressive behavior and the expression of NR1, NR2A receptor in postpartum depression rats induced by maternal separation. Methods Twenty-four Sprague-Dawley female rats were randomly divided into control group( CON group) (n=8),postpartum depression group(PPD group) (n=8),postpartum depression with lactobacillus casei group(PPD+ Lactobacillus casei group) (n=8). Rats of the control group were given no interventions. Rats in PPD group and PPD+Lactobacillus casei group were given maternal separation to establish depression model. And then the rats in PPD+Lactobacillus casei group were treated with Lactobacillus casei ( 8× 108CFU/(kg·d)) for 4 weeks after 14 days of maternal separation. The forced swimming test ( FST) was employed to detect the depressive behaviors. Lactobacillus,Bifidobacterium,Enterococcus faecalis and Esche-richia coli in cecum of rats and the expression of NR1 and NR2A mRNA in hippocampus of rats were detec-ted by real-time fluorescence quantitative PCR. Results Compared with CON group ((157. 50±8. 13) s), the immobility time of PPD group((200. 00±10. 35) s) was significantly longer (t=-3. 23,P<0. 05). Com-pared with PPD group,the immobility time of PPD+ Lactobacillus casei group ((153. 25±7. 41) s) was sig-nificantly shortened (t=3. 67,P<0. 05),and the depressive behavior was improved. Compared with CON group,the mRNA expression of Lactobacillus ((1. 47± 0. 08),t=-5. 87,P<0. 01),Enterococcus faecalis ((1. 23±0. 08),t=-2. 85,P<0. 05) and Escherichia coli( (1. 33±0. 07),t=-4. 96,P<0. 01) in caecum were significantly increased in PPD group, while that of Bifidobacterium decreased significantly (( 0. 65 ± 0. 07),t=5. 18,P<0. 01). Compared with PPD group,the mRNA expression of Lactobacillus ( ( 1. 05 ± 0. 05),t=3. 67,P<0. 01),Enterococcus faecalis ((0. 97±0. 05),t=2. 74,P<0. 05) and Escherichia coli ((1. 06±0. 05),t=3. 31,P<0. 01) were significantly decreased in PPD+ Lactobacillus casei group,while that of Bifidobacterium increased significantly ((0. 98± 0. 04),t=-4. 26,P<0. 01). Compared with CON group,the mRNA expression of NR1 receptor (( 0. 57 ± 0. 04), t=9. 65, P<0. 01) and NR2A receptor ((0. 60±0. 05),t=8. 64,P<0. 05) in hippocampus of rats in PPD group were significantly decreased. Com-pared with PPD group,the expression of NR1 receptor ((1. 01±0. 05),t=-5. 39,P<0. 01) and NR2A re-ceptor ((0. 98±0. 07),t=-3. 91,P<0. 05) in hippocampus were increased significantly in PPD+ Lactoba-cillus casei group. Conclusion Lactobacillus casei can improve the depressive behavior of postpartum depression in rats,and improve the intestinal flora,which affect the expression of NR1 and NR2A in hippocampus.

Increased Nociceptive Responses in Streptozotocin-Induced Diabetic Rats and the Related Expression of Spinal NR2B Subunit of N-Methyl-D-Aspartate Receptors

BACKGROUND: This study investigated the role of NR2B in a modulated pain process in the painful diabetic neuropathy (PDN) rat using various pain stimuli. METHODS: Thirty-two Sprague-Dawley male rats were randomly allocated into four groups (n=8): control, diabetes mellitus (DM) rats and diabetic rats treated with ifenprodil at a lower dose (0.5 µg/day) (I 0.5) or higher dose (1.0 µg/day) (I 1.0). DM was induced by a single injection of streptozotocin at 60 mg/kg on day 0 of experimentation. Diabetic status was assessed on day 3 of the experimentation. The responses on both tactile and thermal stimuli were assessed on day 0 (baseline), day 14 (pre-intervention), and day 22 (post-intervention). Ifenprodil was given intrathecally for 7 days from day 15 until day 21. On day 23, 5% formalin was injected into the rats' hind paw and the nociceptive responses were recorded for 1 hour. The rats were sacrificed 72 hours post-formalin injection and an analysis of the spinal NR2B expression was performed. RESULTS: DM rats showed a significant reduction in pain threshold in response to the tactile and thermal stimuli and higher nociceptive response during the formalin test accompanied by the higher expression of phosphorylated spinal NR2B in both sides of the spinal cord. Ifenprodil treatment for both doses showed anti-allodynic and anti-nociceptive effects with lower expression of phosphorylated and total spinal NR2B. CONCLUSION: We suggest that the pain process in the streptozotocin-induced diabetic rat that has been modulated is associated with the higher phosphorylation of the spinal NR2B expression in the development of PDN, which is similar to other models of neuropathic rats.

Effect of Kidney-nourishing,Blood-activating,Phlegm-resolving and Resuscitation-inducing Decoction on Expression of NMDA Receptor Subtypes in Cochlear Spiral Ganglion Neurons of Tinnitus Rats / 广州中医药大学学报

Objective To observe the effect of Kidney-nourishing, Blood-activating, Phlegm-resolving and Resuscitation-inducing Decoction(KBPRD)on the expression of N-methyl-D-aspartate(NMDA)receptor subtype 1,2A,2B(NR1,NR2A,NR2B)in the cochlear spiral ganglion neurons(SGN)of tinnitus rats and to explore its mechanism, thus to provide experimental evidence for the treatment of tinnitus with KBPRD. Methods Sixty rats were randomly divided into normal group,model group,western medicine group,and low-,middle-,and high-dose Chinese medicine groups, 10 rats in each group. Rats were given intraperitoneal injection of sodium salicylate combined with water deprivation to induce tinnitus model. After successful establishment of the model, the rats in low-,middle-,and high-dose Chinese medicine groups were given gastric administration of KBPRD in the dosage of 5.5, 11, 22 g·kg-1·d-1 respectively, the rats in western medicine group were given gastric administration of 5 mg·kg-1·d-1 of carbamazepine,and rats in the model group and normal group were given gastric administration of 2 mL of normal saline,once every day,treatment time covering 8 weeks. The expression levels of NR1,NR2A,and NR2B in the cochlear SGN was detected by immunoblotting and real-time quantitative reverse transcription-polymerase chain reaction(qRT-PCR)after 8 weeks of treatment. Results Compared with the normal group,the expression levels of NR1, NR2A and NR2B in the model group were increased, the difference being significant (P < 0.05). Compared with the model group,the expression of NR1,NR2A and NR2B in low-,middle-,and high-dose Chinese medicine groups were significantly decreased(P<0.05).Conclusion KBPRD is effective on relieving tinnitus of rats, and its mechanism is correlated with lowering the increased expression of NR1,NR2A and NR2B in SGN of tinnitus rats.

Novel homozygotic mutation in the NR2E3 gene in a family affected with Goldmann-Favre syndrome / 中华眼底病杂志

Objective To identify the pathogenic genes and mutations in a Hui population family with Goldmann-Favre syndrome.Methods A two-generation Hui population family with consanguineous marriage including 4 individuals was enrolled in this study.DNA was extracted from 4 ml peripheral venous blood of all participants.The DNA sequence was performed by Ophthalmology Gene panel sequencing through Ion PGM platform.Then the selected mutations were proved by PCR-Sanger sequencing method.Pathogenic analysis of the mutation was done by means of retrieving PubMed and related databases.And the function of mutation effect was interpreted by protein prediction software.Results The sequence result showed that a novel homozygous mutation in NR2E3,c.925C ＞ T (p.R309W),which resulted in conversion of arginine to tryptophan at position 309 of the photoreceptor-specific retinal nuclear receptor.Parents of the proband were carriers of the heterozygous mutation.The 309 amino acid locus of NR2E3 protein product was highly conserved among species,and protein prediction softwares predicted the mutation as harmful.Conclusion The homozygous mutation c.925C＞T (p.R309W) in NR2E3 cause Goldmann-Favre syndrome in this patient.

In silico analysis of binding interaction of conantokins with NMDA receptors for potential therapeutic use in Alzheimers disease

Abstract Background The N-methyl-D-aspartate (NMDA) receptors are glutamate receptors that play vital roles in central nervous system development and are involved in synaptic plasticity, which is an essential process for learning and memory. The subunit N-methyl D-aspartate receptor subtype 2B (NR2B) is the chief excitatory neurotransmitter receptor in the mammalian brain. Disturbances in the neurotransmission mediated by the NMDA receptor are caused by its overexposure to glutamate neurotransmitter and can be treated by its binding to an antagonist. Among several antagonists, conantokins from cone snails are reported to bind to NMDA receptors. Methods This study was designed to analyze the binding mode of conantokins with NMDA receptors in both humans and rats. To study interactions, dockings were performed using AutoDock 4.2 and their results were further analyzed using various computational tools. Results Detailed analyses revealed that these ligands can bind to active site residues of both receptors as reported in previous studies. Conclusions In light of the present results, we suggest that these conantokins can act as antagonists of those receptors and play an important role in understanding the importance of inhibition of NMDA receptors for treatment of Alzheimers disease.

miR-27b inhibits gastric cancer metastasis by targeting NR2F2

Increasing attention is focused on the down-regulation of miRNAs in cancer process. Nuclear receptor subfamily 2 (NR2F2, also known as COUP-TFII) is involved in the development of many types of cancers, but its role in gastric cancer remains elusive. In this experiment, oncomine and Kaplan-meier database revealed that NR2F2 was up-regulated in gastric cancer and that the high NR2F2 expression contributed to poor survival. MicroRNA-27b was targeted and down-regulated by NR2F2 in human gastric cancer tissues and cells. The ectopic expression of miR-27b inhibited gastric cancer cell proliferation and tumor growth in vitro and in vivo. Assays suggested that the overexpression of miR-27b could promote MGC-803 cells' migration and invasion and retard their metastasis to the liver. In addition, down-regulation of miR-27b enhanced GES-1 cells' proliferation and metastasis in vitro. These findings reveal that miR-27b is a tumor suppressor in gastric cancer and a biomarker for improving patients' survival.

In silico analysis of binding interaction of conantokins with NMDA receptors for potential therapeutic use in Alzheimer's disease

Background The N-methyl-D-aspartate (NMDA) receptors are glutamate receptors that play vital roles in central nervous system development and are involved in synaptic plasticity, which is an essential process for learning and memory. The subunit N-methyl D-aspartate receptor subtype 2B (NR2B) is the chief excitatory neurotransmitter receptor in the mammalian brain. Disturbances in the neurotransmission mediated by the NMDA receptor are caused by its overexposure to glutamate neurotransmitter and can be treated by its binding to an antagonist. Among several antagonists, conantokins from cone snails are reported to bind to NMDA receptors. Methods This study was designed to analyze the binding mode of conantokins with NMDA receptors in both humans and rats. To study interactions, dockings were performed using AutoDock 4.2 and their results were further analyzed using various computational tools. Results Detailed analyses revealed that these ligands can bind to active site residues of both receptors as reported in previous studies. Conclusions In light of the present results, we suggest that these conantokins can act as antagonists of those receptors and play an important role in understanding the importance of inhibition of NMDA receptors for treatment of Alzheimer's disease.(AU)

Nr2e1 Downregulation Is Involved in Excess Retinoic Acid-induced Developmental Abnormality in the Mouse Brain / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>This study aimed to investigate the expression pattern and function of Nuclear receptor subfamily 2 group E member 1 (Nr2e1) in retinoic acid (RA)-induced brain abnormality.</p><p><b>METHODS</b>The mouse model of brain abnormality was established by administering 28 mg/kg RA, and neural stem cells (NSCs) were isolated from the mouse embryo and cultured in vitro. Nr2e1 expression was detected by whole mount in situ hybridization, RT-PCR, and Western blotting. Nr2e1 function was determined by transducing Nr2e1 shRNA into NSCs, and the effect on the sonic hedgehog (Shh) signaling pathway was assessed in the cells. In addition, the regulation of Nr2e1 expression by RA was also determined in vitro.</p><p><b>RESULTS</b>Nr2e1 expression was significantly downregulated in the brain and NSCs of RA-treated mouse embryos, and knockdown of Nr2e1 affected the proliferation of NSCs in vitro. In addition, a similar expression pattern of Nr2e1 and RA receptor (RAR) α was observed after treatment of NSCs with different concentrations of RA.</p><p><b>CONCLUSION</b>Our study demonstrated that Nr2e1 could be regulated by RA, which would aid a better understanding of the mechanism underlying RA-induced brain abnormality.</p>

Cell-penetrating peptdie based on peptide array technique inhibiting NR2B phosphorylation in inflammato-ry pain rats / 实用医学杂志

Objective We used peptide array technique to construct a peptide FynP inhibiting the interac-tion between Fyn and PSD95 in vitro therefor with a potential for inbiting NR2B phosphorylation level(p-NR2B). This experiment was designed to examine whether FynP(deliverd with TAT-LK15)can inhibit interaction between Fyn and PSD95 in inflammatory pain rats,and therefore inhibit NR2B phosphorylationin in vivo. Methods TAT-LK15 was complexed with FynP(cell-penetrating peptide Tat-LK15/FynP)or scrambled control FynP(Tat-LK15/mFynP). Changes of p-NR2B were detected using western-blot in SCDH of chronic inflammatory pain rats following intraperitoneal injection of Tat-LK15/FynP,meanwhile,the effect of Tat-LK15/FynP on the interaction between Fyn and PSD-95 was tested by co-immunoprecipitation. Pain control efficacy was evaluated by changes of mechani-cal withdrawal threshold(MWT)and thermal withdrawal duration(TWL)in these rats. Results Interaction be-tween Fyn and PSD-95 was efficiently inhibited by intraperitoneal injection of TAT-LK15/FynP complexes while in-jection of FynP or TAT-LK15/mFynP complexes did not show this inhibitory effect. NR2B phosphorylation level was also inhibited by injection of TAT-LK15/FynP,and the changes of p-NR2B levels were reduced by 52%compared to chronic inflammatory pain rats without treatment. FynP or TAT-LK15/mFynP did not show this effect. Moreover, injection of TAT-LK15/FynP complexes significantly reduced MWT and increased TWL of chronic inflammatory pain rats accordingly. Conclusion FynP delivered by Tat-LK15 can perturb Fyn and PSD95 interaction and then inhibit NR2B phosphorylation activation therfor relieve chronic inflammatory pain.