Opioid medications have been used for pain management, but there are negative side effects, including a potential delay in recovery and increased risk of permanent disability. In this paper, analgesic targets including N-methyl-D-aspartate receptors, cannabinoid receptors, prostaglandin E2 receptor 4 receptors, matrix metalloproteinase receptors, and some new targets for blocking pain signaling pathways associated with these receptors are reviewed. In addition, some novel agonists, antagonists, and leading compounds with agonistic (antagonistic) activities interacting with the target are also described. These novel compounds usually have better analgesic activity and lower side effects than traditional opioids. They are expected to be developed into new analgesics and benefit clin?ical patients who need pain treatment.

OBJECTIVE To explore the protective effect and mechanism of ophiopogonin D (OP-D) on oxidative stress injury of H9c2 cells induced by H2O2. METHODS An oxidative damage model of H9c2 cells was established by H2O2 induction. The cells were divided into control group (cultured in serum-free medium for 28 h), H2O2 injury model group (treated with H2O2400μmol·L-1 for 3 h), OP-D 5, 10 and 20 μmol · L-1 pretreatment groups (treated with H2O2400 μmol · L-1 for 3 h after OP-D pretreat?ment for 24 h), and an inhibitor of CYP2J3, 6-(2-proparglyloxyphenyl) hexanoic acid (PPOH) group (OP-D 20μmol·L-1+PPOH 10μmol·L-1, PPOH was added to the cells 1 h before OP-D treatment). Cell activity was measured by MTT method, levels of dihydroxyeicosatrienoic acid (11,12-DHET and 14,15-DHET respectively) were detected by enzyme-linked immunosorbent assay (ELISA), while levels of malondialdehyde (MDA), nitric oxide (NO) and activity of lactate dehydrogenase (LDH), superoxide dismutase (SOD) were detected by assay kits. Flow cytometry (FCM) was used to detect reactive oxygen species (ROS) and apoptosis. Western blotting was used to detect the expressions of CYP2J3, Akt phos?phorylation (p-Akt) protein and endothelial nitric oxide synthase phosphorylation (p-eNOS) protein in cells, and the possible mechanism by which OP-D reduces oxidative stress was further verified with PPOH. RESULTS H2O2400μmol · L-1 significantly inhibited H9c2 cell viability (P<0.01), and OP-D signifi?cantly increased the cell survival rate after H2O2 injury (P<0.01). Different concentrations of OP-D increased the level of 11,12- DHET and 14,15-DHET (P<0.05, P<0.01). OP-D increased the level of NO in cells after H2O2-induced injury (P<0.05, P<0.01), enhanced the activity of SOD (P<0.05), and decreased the level of MDA and LDH (P<0.05, P<0.01). OP-D significantly reduced oxidative stress and apoptosis after H2O2 injury (P<0.05, P<0.01). OP-D pretreatment increased the protein and mRNA expression of CYP2J3 (P<0.05, P<0.01) and the phosphorylation of PI3K/Akt-eNOS pathway after H2O2 injury (P<0.05, P<0.01). After PPOH was given in advance, the protective effect of OP-D was inhibited (P<0.05, P<0.01). CONCLUSION OP-D can reduce H2O2-induced H9c2 cell damage, which may be related to the activation of PI3K pathway and the phosphorylation of its downstream factors Akt and eNOS by inducing CYP2J3 expression and increasing the contents of 11,12-DHET and 14,15-DHET.

OBJECTIVE To evaluate the genotoxicity of naproxen (NPX) impurities acetylnerolin (Ace). METHODS The genotoxicity of Ace was predicted by ADMET, Derek and Sarah with the quanti?tative structure-activity relationship (QSAR). The chromosomal aberration and bacterial reverse-muta?tion (Ames) tests were performed to verify the above results. In chromosomal aberration tests, CHL cells were incubated with Ace 10, 20 and 40 mg · L-1 for 4 h in the presence or absence of metabolic activation system solution (S9 mix). Methyl methane sulfonate (MMS) 20 mL · L-1 without S9 mix and cyclophosphamide (CP) 12 mg · L-1 with S9 mix served as positive control. The number of chromo?somes in each aberrant metaphase (including fissure, exchange, ring, break and polyploid) was counted and recorded, when the distortion rate less than 5％was considered negative and more than 10％was considered positive. In Ames test, the potential mutagenicity was evaluated using five strains of S. typhimurium ( TA97,TA98,TA100,TA102 and TA1535). They were treated with Ace 5, 25, 125 and 625μg per plate with or without S9 mix and incubated for 48-72 h. When without S9 mix, Dexon 50μg per plate served as positive control for TA97 and TA98, MMS 2.0μL per plate served as positive control for TA100 and TA102, and sodium azide 1.5μg per plate served as positive control for TA1535. When with S9 mix, 2-AF 100 μg per plate served as positive control for TA97, TA98 and TA100, 1, 8-dihydroxyanthraquinone (100μg per plate) served as positive control for TA102 and CP 50μg per plate served as positive control for TA1525. When the number of colonies was at least two-fold that of the negative control, the compound was considered mutagenic. RESULTS Although the Derek and Sarah software predicted that the NPX impurities were not genotoxic, ADMET data showed that Ace could induce chromosomal aberrations. The distortion rate of Ace 40 mg · L-1 was greater than 5％, but less than 10％. The distortion rate of Ace was less than 5％when<20 mg·L-1. Consistent with the results of ADMET, Ace might induce chromosomal aberrations. Ames test results showed that Ace did not signifi?cantly increase the number of bacteria (5-625μg per plate) compared with the negative control. Contrary to the ADMET results, Ace had no mutagenicity. CONCLUSION Ace has potential chromosomal muta?genicity. For life-long usage of NPX, the content of Ace should be reduced from 0.15％of conventional impurities to 0.015％.

Epilepsy is a disorder of the brain charac-terized by abnormal neuron excitability.However,the underlying molecular mechanism of neuron excitability modulation remains elusive.With the help of bioinformatic methods,we have identified receptor-type tyrosine-pro-tein phosphatase-like N(PTPRN)as a critical gene dur-ing epileptogenesis.PTPRN recruits NEDD4L ubiquitin E3 ligase to NaV1.2 sodium channels,facilitating NEDD4L-mediated ubiquitination and endocytosis.Knockout of PTPRN endows hippocampal granule cells with augmented depolarization currents and higher intrinsic excitability,which is reflected by increased seizure susceptibility of transgenic mice.On the contrary,reduced neuron excit-ability and decreased seizure susceptibility are observed after PTPRN overexpression.Meanwhile,we find that a 133 aa fragment recaptures modulation effect of PTPRN full-length,and this fragment shows therapeutic potential towards epilepsy caused by NaV1.2 gain of function vari-ants.In brief,our results demonstrate PTPRN playsa criti-calroleinregulatingneuronexcitability,providing a poten-tial therapeutic approach for epilepsy.

OBJECTIVE Epileptic networks are char-acterized as two states,seizures or more prolonged inter-ictal periods.However,cellular mechanisms underlying the contribution of interictal periods to ictal events remain unclear.METHODS Here,we present the procedure for labeling seizure-activated and interictal-activated neuro-nal ensembles in mouse hippocampal kindling model using an enhanced-synaptic-activity-responsive element.This technique is combined with genetically encoded effectors to characterize and manipulate neuronal ensembles recruited by focal seizures(FS-Ens)and interictal periods(IP-Ens)in piriform cortex,a region that plays a key role in seizure generation.RESULTS Ca2+ activities and histo-logical evidence reveal a disjointed correlation between the two ensembles during FS dynamics.Optogenetic acti-vation of FS-Ens promotes further seizure development,while IP-Ens protects against it.Interestingly,both ensem-bles are functionally involved in generalized seizures(GS)due to circuit rearrangement.IP-Ens bidirectionally modulates FS but not GS by controlling coherence with hippocampus.CONCLUSION This study indicates that the interictal state may represent a seizure-preventing environment,and the interictal-activated ensemble may serve as a potential therapeutic target for epilepsy.

OBJECTIVE Cognitive deficit is a com-mon comorbidity in temporal lobe epilepsy(TLE)and that is not well controlled by current therapeutics.Currently,how epileptic seizure affects cognitive performance remains largely unclear.The subiculum is the major out-put of the hippocampus,which projects to entorhinal cor-tex and other more distinct brain regions.Physiologically,the subiculum codes spatial working memory and naviga-tion information including place,speed,and trajectory.Importantly,prior studies have noted the importance of the subiculum in the beginning,spreading,and generaliz-ing process of hippocampal seizure.How seizure-activated neurons in subiculum participate in cognitive impairment remains largely elusive.METHODS In this study,we sought to label the subicular seizure-activated c-fos+ neu-rons with a special promoter with enhanced synaptic activity-responsive element E-SARE in the subiculum,combined with chemogenetics and designer receptors exclusively activated by designer drugs(DREADDs),Ca2+ fiber photometry approaches,and behavioral tasks,to reveal the role of these neurons in cognitive impairment in epilepsy.RESULTS We found that chemogenetic inhibi-tion of subicular seizure-tagged c-fos+ neurons(mainly CaMK Ⅱ α+ glutamatergic neurons)alleviates seizure generalization and improves cognitive performance in the hippocampal CA3 kindling TLE model.While inhibition of seizure-labeled c-fos+ GABAergic interneuron shows no effect on seizure and cognition.As a comparison,che-mogenetic inhibition of the whole subicular CaMK Ⅱ α+ neuron impairs cognitive function in na?ve mice in basal condition.Notably,inhibition of subicular seizure-tagged c-fos+ neurons enhances the recruitment of cognition-responsive c-fos+ neurons via increasing neural excitability during cognition tasks.CONCLUSION Our results dem-onstrate that subicular seizure-activated c-fos+ neurons contribute to cognitive impairment in TLE,suggesting sei-zure-tagged c-fos+ neurons as the potential therapeutic target to alleviate cognitive impairment in TLE.

OBJECTIVE Temporal lobe epilepsy is a common neurological disease caused by abnormal syn-chronized discharge in the brain and it is mainly treated through long-term use of anti-epileptic drugs(AEDs).This project is supposed to provide an electro-responsive and brain-targeted drug delivery system(DDS)for on-demand drug release,which could promptly block the transmis-sion of epileptic discharges.METHODS The DDS was fab-ricated by co-polymerization of dopamine and pyrrole,together with conjugation of brain-targeted peptide.A number of characterization including electron microscopy,thermogravimetric analysis,dynamic light scattering and other methods were conducted to evaluate the physio-chemical properties of the nanomaterials.In vitro study based on a home-made electric device and high perfor-mance liquid chromatography was performed to record drug release profiles.Three epileptic models including acute,continuous and spontaneous models were estab-lished for the evaluation of therapeutic efficacy.RESULTS Our polymeric DDS has a nanoscale size(ca.80 nm)and could load AEDs such as phenytoin(drug loading capacity 20.4%).The hybrid nanomaterials can improve the brain delivery efficiency through a combination of receptor-mediated transcytosis and near-infrared-enabled brain transport.In vitro study proved that the DDS could release phenytoin in the electric field in a sensitive(50 μA),quick(30 s)and sustained(>3 times)manner.In vivo study demonstrated excellent anti-epileptic effects in a lower dose(20%).Biosafety study further verified that our strategy has limited damage.CONCLUSION For on-demand seizure control,we have developed a nano-engineered DDS with the capability of electro-responsive drug release and brain-targeted accumula-tion.The DDS could increase the AEDs accumulation at epileptic region and release the AEDs in response to the epileptic discharges.Such strategy could timely inhib-it the epileptic seizure.Our work provides a promising approach to"smart"therapy of epilepsy and sheds light on development of pharmacotherapy of other brain disorders.

OBJECTIVE Epilepsy is considered a cir-cuit-level dysfunction associated with imbalanced excita-tion-inhibition,it is therapeutically necessary to identify key brain regions and related circuits in epilepsy.The subic-ulum is an essential participant in epileptic seizures,but the circuit mechanism underlying its role remains largely elusive.METHODS Here we deconstruct the diversity of subicular circuits in mouse models of epilepsy.Fiber pho-tometry was used to detect intrinsic activities of subicular PV,SST-positive interneurons and CaMK Ⅱ α-positive pyramidal neurons.Optogenetics and chemogenetics were used to selectively active or inactive subicular neu-rons or their projecting terminals.We also used in vivo and in vitro electrophysiology to record membrane charac-teristics of single neuron in distinct sub-regions of the subiculum.Finally,single pulse test was used to detect synaptic transmission strength between the subiculum and its downstream target.RESULTS First,we found that two majority of subicular interneurons,which inner-vate local pyramidal neurons to constrain their excitability,PV and SST-positive neurons showcase distinct calcium dynamics during hippocampal seizures.This could be attributed to distinct neural inputs from para-hippocampal regions of these two neuronal types.During epileptogen-esis,PV and SST neurons undergo different circuit reor-ganization patterns,that is,remarkable increase of exter-nal input to subicular PV neurons are seen after seizures,while SST cells receive decimated neural input.As their downstream targets,excitatory subicular pyramidal neu-rons are also intrinsically activated during hippocampal seizures.Moreover,we found that the subiculum hetero-geneously controls the generalization of hippocampal sei-zures by projecting to different downstream regions.No-tably,anterior thalamus projecting subicular neurons bidi-rectionally mediate seizures,while entorhinal cortex-pro-jecting subicular neurons act oppositely in seizure modu-lation.These two subpopulations are structurally and functionally dissociable.An intrinsically enhanced hyper-polarization-activated current and robust bursting intensity in anterior thalamus-projecting neurons facilitate synaptic transmission,thus contributing to the generalization of hippocampal seizures.CONCLUSION These results demonstrate that subicular neurons and circuits have diverse roles in epilepsy,suggesting the necessity to pre-cisely target specific subicular circuits for effective treat-ment of epilepsy.

OBJECTIVE To reveal the role of the basal forebrain(BF)GABAergic neurons in the regulation of isoflurane anesthesia and to elucidate the underlying neural pathways.METHODS The activity of BF GABAer-gic neurons was monitored during isoflurane anesthesia using a genetically encoded calcium indicator in Vgat-Cre mice of both sexes.The activity of BF GABAer-gic neurons was manipulated by chemogenetic and opto-genetic approaches.Sensitivity,induction time and emer-gence time of isoflurane anesthesia were estimated by righting reflex.The electroencephalogram(EEG)power and burst-suppression were monitored by EEG recording.The effects of activation of GABAergic BF-thalamic reticu-lar nucleus(TRN)pathway on isoflurane anesthesia were investigated with optogenetics.RESULTS The activity of BF GABAergic neurons was generally inhibited during isoflurane anesthesia,obviously decreased during the induction of anesthesia and gradually restored during the emergence from anesthesia.Activation of BF GABAergic neurons with chemogenetics and optogenetics promoted behavioral emergence from isoflurane anesthesia,with decreased sensitivity to isoflurane,delayed induction and accelerated emergence from isoflurane anesthesia.Optogenetic activation of BF GABAergic neurons prom-oted cortical activity during isoflurane anesthesia,with decreased EEG delta power and burst suppression ratio during 0.8%and 1.4%isoflurane anesthesia,respectively.Similar to the effects of activating BF GABAergic cell bod-ies,photostimulation of BF GABAergic terminals in the TRN also strongly promoted cortical activation and behav-ioral emergence from isoflurane anesthesia.CONCLU-SION The GABAergic neurons in the BF is a key neural substrate for general anesthesia regulation that facilitates behavioral and cortical emergence from general anesthe-sia via the BF-TRN pathway.

Sleep is essential for the maintenance of human normal functions.Nowadays,the occurrence of active sleep deprivation(ASD)or passive sleep depriva-tion(PSD)is becoming more and more common due to the inability of the body adapting to the rapid changes in the internal and external environment.SD is not only an action,a brief process or a result,but also a directly or indirectly sustained state,which is associated to sleep time,circadian rhythm or sleep quality.SD can lead to numerous adverse effects on the body,such as sleep-related acute and chronic diseases.Long-term SD increases the risk of neurological and cardiovascular dis-eases as well as immune system dysfunction.In addi-tion,SD may affect the reproductive health of the body,giving rise to a series of potential fertility problems.In recent years,the correlation research and mechanism between SD and the related diseases have become a focus of scholars' attention.Numerous lines of evidence suggest that pathological sleep,such as insomnia and sleep apnea syndrome,is associated with impaired repro-ductive function.Disruptions in the circadian rhythm can also lead to impaired hypothalamic-pituitary-gonadal(HPG)axis function and thereby interfere with the repro-ductive process.Our research team has demonstrated that SD significantly affects the fertility of male and female rats and has adverse effects on reproduction.By new generation sequencing(NGS)and RT-PCR verifica-tion,we have identified differently expressed genes that are involved in mediating the effect of SD on fertility.However,the mechanisms and biological macromolecules regulated by SD are worthy of being further explored.This paper provides a brief review of SD research and then focuses on the adverse impact of SD on fertility,conducting a literature review to sort out the ideas and pro-vide references for research in this field.