OBJECTIVES:To investigate the mechanism by which the pyramidal neurons of the anterior cingulate cortex (ACC) modulate the effects of enriched environment (EE) for relieving anxiety-like behaviors in mice. METHODS:C57BL/6J mice were randomly divided into control group, restraint stress (RS) group, and RS+EE group (n=8). The mice in the latter two groups were subjected to RS for 2 h daily for 3 days, and those in RS+EE group were housed in an EE during modeling. Anxiety-like behaviors of the mice were evaluated using the elevated plus-maze tests (EPM) and open field test (OFT). Changes in c-Fos expression in the ACC of the mice were detected with immunofluorescence assay, and pyramidal neuron excitability in the ACC (PynACC) was measured using patch-clamp technique. The miniature excitatory and inhibitory postsynaptic currents (mEPSC and mIPSC, respectively) were analyzed to assess synaptic transmission changes. RESULTS:Behavioral tests showed obvious anxiety-like behaviors in RS mice, and such behavioral changes were significantly improved in RS+EE mice. Immunofluorescence staining revealed significantly increased c-Fos expression in the ACC in RS mice but lowered c-Fos expression in RS+EE group. Compared with the control mice, the RS mice showed increased action potential firing rate of PynACC, which was significantly reduced in RS+EE group. Compared with the RS mice, the RS+EE mice showed also decreased frequency of mEPSCs of PynACC, but the amplitude exhibited no significant changes. No obvious changes in the frequency or amplitude of mIPSCs were observed in RS+EE mice. CONCLUSIONS:EE reduces excitability of PynACC to alleviate anxiety-like behaviors induced by RS in mice.

OBJECTIVE:To elucidate the modulation mechanism of Suanzaoren Decoction (SZRD) on basolateral amygdala (BLA) neuronal activity to alleviate chronic restraint stress (CRS)-related behavioral deficits. METHODS:The male C57BL/6J mice were assigned to 4 groups using the complete randomization method, including control (CON, n=19), CRS (n=19), SZRD (n=21), and fluoxetine (Flu, n=22) groups. Mice were restrained for 6 h per day, over a 21-d period to establish CRS models. The CON group remained in their cages without food or water during the 6-h matching period. SZRD and Flu groups received intragastric administration of SZRD (4.68 g/kg) and Flu (20 mg/kg) daily, respectively, 30 min before restraint for 21 consecutive days. The therapeutic effects of SZRD were evaluated using behavioral tests including the tail suspension test, elevated plus maze test, and forced swimming test. The cellular Fletcher B. Judson murine osteosarcoma proto-oncogene (c-Fos) expression in the BLA was measured using immunofluorescence, while action potential (AP) firing and synaptic transmission in BLA pyramidal neurons were evaluated using whole-cell patch-clamp recordings. RESULTS:SZRD administration significantly increased time spent in the open arms and open-arm entries while reducing immobility time (P<0.05 or P<0.01). It downregulated CRS-induced c-Fos expression and AP firing of pyramidal neurons in the BLA (P<0.01). Additionally, SZRD selectively attenuated excitatory (P<0.01), but not inhibitory, synaptic transmission onto BLA pyramidal neurons. CONCLUSION:SZRD alleviated CRS-induced anxiety- and depression-like behaviors in mice by modulating the excitability and synaptic transmission of BLA pyramidal neurons.

The purpose of this study was to investigate the anxiety-like behaviors, circadian rhythms and sleep, and to elucidate the possible underlying mechanisms of the abnormal sleep behavior in Shank3 gene knockout (Shank3-KO) mice. The anxiety-like behaviors were detected by elevated plus-maze (EPM) test, open field test (OFT) and tail suspension test (TST). The circadian rhythms were detected by running wheel test. The electroencephalogram (EEG)/electromyogram (EMG) recordings were performed synchronically by polysomnograph. The distribution of SHANK3 in anterior cingulate cortex (ACC), paraventricular thalamus (PVT), nucleus accumbens (NAc), basolateral amygdala (BLA) and hippocampal CA2 region in wild type (WT) mice was detected by immunofluorescence assay. The protein expression of c-Fos in PVT, ACC and NAc was also detected by immunofluorescence assay during light cycle. The colocalization of c-Fos and vesicular glutamate transporter 2 (Vglut2, a marker for glutamatergic neurons) in the PVT was detected by immunofluorescence double labeling experiment. The results of EPM test showed that, compared with the WT mice, the Shank3-KO mice showed less time in open arms and less number of open arm entries. The results of OFT showed that the Shank3-KO mice showed less time in central area and less number of central area entries. The immobility time of Shank3-KO mice was increased in the TST. The results of running wheel rhythm test showed that the phase shift time of Shank3-KO mice in the continuous dark period was increased. The results of EEG/EMG recording showed that, compared with the WT mice, the duration of wakefulness in Shank3-KO mice was increased and the duration of non-rapid eye movement (NREM) sleep was decreased during light phase; The bout number of wakefulness was increased, the bout number of NREM sleep was decreased, NREM-wake transitions were increased, and wake-NREM transitions were decreased during light phase. SHANK3 was expressed in ACC, PVT, NAc and BLA in the WT mice. The expression of c-Fos in the PVT of Shank3-KO mice was up-regulated 2 h after entering the light phase, and majority of c-Fos was co-localized with Vglut2. These results suggest that the anxiety level of Shank3-KO mice is increased, the regulation of the internal rhythms is decreased, and the bout number of wakefulness is increased during light phase. The glutamatergic neurons in PVT may be involved in the regulation of abnormal sleep behavior in Shank3-KO mice during the light phase.

Nav1.3 is a tetrodotoxin-sensitive voltage-gated sodium channel isoform encoded by SCN3A, the abnormal expression of which plays a crucial role in the generation of ectopic discharge, as well as being associated with allodynia and hyperalgesia. Using bioinformatics analysis, we showed that miR-30b-5p directly targets SCN3A. We aimed to explore whether miR-30b-5p can participate in trigeminal neuralgia (TN) in rats by regulating the expression of Nav1.3. The rat TN model was constructed through infraorbital nerve-chronic constriction injury (ION-CCI), which was verified by measuring the change in mechanical threshold and the expression of activating transcription factor 3 (a marker of nerve damage) in the trigeminal ganglia (TG). The expression of miR-30b-5p in postoperative TG was downregulated, whereas that of Nav1.3 was upregulated in rats subjected to ION-CCI. Overexpression of miR-30b-5p repressed the expression of Nav1.3 in TG and alleviated ION-CCI-induced TN. MiR-30b-5p targets to regulate the expression of SCN3A, thereby reducing or aggravating TN. Therefore, miR-30b-5p may be a novel therapeutic target for neuropathic pain.

Blue light has been shown to impair spatial memory in mice. However, the underlying neural circuit mechanisms of this impairment remain elusive. Our study revealed that 600-lux blue light disrupted spatial memory retrieval in male mice through the secondary visual cortex (V2M) → the prelimbic cortex (PrL) neural pathway. Specifically, viral tracing and multichannel fiber optic recordings demonstrated that glutamatergic V2M neurons (V2MGlu) projected to GABAergic neurons in the PrL (PrLGABA), thereby inhibiting the excitability of glutamatergic PrL neurons (V2MGlu→PrLGABA→Glu). The selective ablation of V2MGlu neurons projecting to the PrL eliminated the detrimental effects of blue light on spatial memory retrieval. Furthermore, optogenetic or chemogenetic activation of the V2MGlu → PrLGABA→Glu neural circuitry replicated the impairing effects of blue light on spatial memory in mice. Conversely, inhibition of the V2MGlu → PrLGABA→Glu neural circuitry reversed the spatial memory damage induced by blue light. Collectively, our findings elucidate the V2MGlu → PrLGABA→Glu neural circuitry as a potential mediator of blue-light-induced spatial memory impairment in male mice. A V2MGlu→PrLGABA→Glu circuit underlies blue light-induced spatial memory impairment in mice.

The regulation of circadian rhythms and the sleep–wake states involves in multiple neural circuits. The suprachiasmatic nucleus (SCN) is a circadian pacemaker that controls the rhythmic oscillation of mammalian behaviors. The basal forebrain (BF) is a critical brain region of sleep–wake regulation, which is the downstream of the SCN. Retrograde tracing of cholera toxin subunit B showed a direct projection from the SCN to the horizontal limbs of diagonal band (HDB), a subregion of the BF. However, the underlying function of the SCN–HDB pathway remains poorly understood. Herein, activation of this pathway significantly increased non–rapid eye movement (NREM) sleep during the dark phase by using optogenetic recordings. Moreover, activation of this pathway significantly induced NREM sleep during the dark phase for first 4 h by using chemogenetic methods. Taken together, these findings reveal that the SCN–HDB pathway participates in NREM sleep regulation and provides direct evidence of a novel SCN-related pathway involved in sleep–wake states regulation.

This study aimed to investigate the mechanism of Dexmedetomidine (DEX) on sleep homeostasis. We will focus on the effect of DEX on the activity and the signal transmission of oxytocin-expressing (Oxytocin, OXT) neurons in the paraventricular nucleus (PVN) of the hypothalamus (PVN OXT ) in the regulation of sleep-wakefulness cycle. In this study, the guide cannula was implanted to microinject DEX into PVN. PVN OXT neurons were specifically activated by the chemogenetic method. Differences in sleep-wakefulness states were monitored by electroencephalogram (EEG)/ electromyogram (EMG) recording. In addition, the effect of DEX on the electrophysiological activity of PVN OXT neurons was examined by whole-cell patch-clamp technique. EEG/EMG results showed that microinjection of DEX in PVN significantly increased the duration of nonrapid eye movement (NREM) sleep in mice. Chemogenetic activation of PVN OXT neurons after internal cannula injection of DEX to PVN increased the amount of wake. Electrophysiological results show that DEX could inhibit the frequency of action potential (AP) and the spontaneous excitatory postsynaptic current (sEPSC) in PVN OXT neurons through α 2 -adrenoceptors. DEX maintained sleep homeostasis by inhibiting excitatory synaptic signaling in PVN OXT neurons through α 2 -adrenoceptors.

Dexmedetomidine (DexM), a highly selective α2-adrenoceptor agonist, significantly reduces postoperative adverse effects, including sleep and circadian rhythm disruptions. Vasoactive intestinal peptide neurons in the suprachiasmatic nucleus (SCNVIP) regulate the synchronization of circadian rhythms with the external environment in mammals. We investigate the effects of DexM on sleep and circadian rhythms, as well as the underlying mechanisms. Using electrophysiological and chemogenetic methods, along with locomotor activity and electroencephalogram/electromyogram recordings, we found that DexM accelerates the rate of re-entrainment following an 8-hour phase advance in the 12-hour light:12-hour dark cycle, increases the amount of non-rapid eye movement sleep, and decreases the mean duration of rapid eye movement sleep. Chemogenetic inhibition of SCNVIP neurons hinders the acceleration of re-entrainment and the changes in the sleep-wakefulness cycle induced by DexM. Electrophysiological results show that DexM increases the firing rate and the frequency of spontaneous glutamatergic postsynaptic currents while decreasing the frequency of spontaneous GABAergic PSCs in SCNVIP neurons through the α2-adrenergic receptor. Additionally, DexM reduces the frequency of miniature GABAergic PSCs in SCNVIP neurons. In conclusion, these findings suggest that DexM promotes sleep and maintains the coordination of circadian rhythms with the external environment by activating SCNVIP neurons through the α2-adrenoceptor.

BackgroundTrigeminal neuralgia (TN) is a common and difficult-to-treat neuropathic pain disorder in clinical practice. Previous studies have shown that Toll-like receptor 4 (TLR4) modulates the activation of the NF-κB pathway to affect neuropathic pain in rats. Voltage-gated sodium channels (VGSCs) are known to play an important role in neuropathic pain electrical activity.ObjectiveTo investigate whether TLR4 can regulate Nav1.3 through the TRAF6/NF-κB p65 pathway after infraorbital nerve chronic constriction injury (ION-CCI).Study design: ION-CCI modeling was performed on SD (Sprague Dawley) rats. To verify the success of the modeling, we need to detect the mechanical pain threshold and ATF3. Then, detecting the expression of TLR4, TRAF6, NF-κB p65, p-p65, and Nav1.3 in rat TG. Subsequently, investigate the role of TLR4/TRAF6/NF-κB pathway in ION-CCI model by intrathecal injections of LPS-rs (TLR4 antagonist), C25-140 (TRAF6 inhibitor), and PDTC (NF-κB p65 inhibitor).ResultsION-CCI surgery decreased the mechanical pain threshold of rats and increased the expression of ATF3, TLR4, TRAF6, NF-κB p-p65 and Nav1.3, but there was no difference in NF-κB p65 expression. After inject antagonist or inhibitor of the TLR4/TRAF6/NF-κB pathway, the expression of Nav1.3 was decreased and mechanical pain threshold was increased.ConclusionIn the rat model of ION-CCI, TLR4 in the rat trigeminal ganglion regulates Nav1.3 through the TRAF6/NF-κB p65 pathway, and TLR4 antagonist alleviates neuropathic pain in ION-CCI rats.

目的 探究短时程光照对视交叉上核(SCN)和外侧缰核(LHB)谷氨酸受体表达的影响.方法 利用T7(3.5 h/3.5 h光暗循环)建立短时程光周期模型组和T24(12 h/12 h光暗循环)正常光建立对照组,通过蛋白免疫印迹(Western blot)的方法对SCN和LHB脑区关键受体蛋白的表达进行分析,包括谷氨酸能受体、垂体腺苷酸环化酶激活肽(PACAP)受体以及下游信号分子.结果 Western blot结果显示,与T24相比,T7下SCN在授时因子时间(ZT)1、ZT 5的α-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体亚型2(GluR2)的表达增多(P<0.05,P<0.01),T7组GluR2总体表达比T24组增多(P<0.01),而LHB中T7组GluR2总体表达比T24组减少(P<0.01),与T24光周期相比,T7光周期下SCN的N-甲基-D-天冬氨酸受体亚型2(NR2B)的总体表达上升(P<0.05),T7光周期下LHB的磷酸化的细胞外调蛋白激酶(P-ERK)的总体表达上升(P<0.05).结论 T7光周期会导致SCN中GluR2和NR2B表达增多、LHB中GluR2受体表达降低及P-ERK的表达升高.

The unique sedative activities with rapid arousal of dexmedetomidine (Dex) are not fully understood. Growing evidence suggests the involvement of the ventrolateral preoptic area (VLPO) in sleep-wake cycle. The major type in the VLPO is sleep-active neurons, inhibited by noradrenaline (NA(-) neurons). The other type of neurons is activated by NA (NA(+) neurons), which are wake-active. Previous research showed that Dex-induced sedation and sleep homeostasis likely share common mechanisms. To explore the underlying mechanisms of Dex in the VLPO, in vivo polysomnography recording and in vitro electrophysiological recording were used in our study. Bath application of Dex (2 & mu;M) increased the firing rate of both VLPO NA(-) and NA(+) neurons. Compared to the control group, there was no difference in the firing rate of both VLPO NA(-) and NA(+) neurons after Dex (2 & mu;M) and RS79948 (1 mM) administration, an & alpha;(2) receptor antagonist. No difference was detected regarding resting membrane potential (RMP) amplitude of both VLPO NA (-) and NA(+) neurons after application of Dex (2 & mu;M). Moreover, Dex (2 & mu;M) significantly reduced the frequency of miniature inhibitory postsynaptic currents (mIPSCs) in both VLPO NA(-) and NA(+) neurons. These electrophysiology results were consistent with behavioral sedation, with increased nonrapid eye movement sleep (NREM sleep) and increased expression of c-Fos in the VLPO during the dark phase after intraperitoneal injection with Dex (80 & mu;g/kg). In conclusion, Dex activates NA(-) and NA(+) neurons in the VLPO via presynaptic & alpha;(2) receptors. This mechanism may explain the unique sedative properties with rapid arousal.Summary StatementDexmedetomidine is an important ICU sedative. The mechanism of dexmedetomidine is not fully understood. Activating NA(-) and NA(+) neurons in the VLPO by dexmedetomidine using polysomnography and electrophysiological recording, this may explain the unique sedative properties with rapid arousal.

Objective:Isoliquiritigenin (ILTG) is a chalcone compound that exhibits hypnotic effects via gamma-aminobutyric acid type A (GABAA) receptors. The ventrolateral preoptic area (VLPO) is a sleep-promoting center that contains a large number of GABA-releasing cells. There are two cell types in the VLPO: one generates a low-threshold spike (LTS), whereas the other lacks an LTS (non-LTS). Method:Whole-cell patch-clamp technology was used to detect the firing and currents of LTS and non-LTS cells in the VLPO. Results:Bath administration of ILTG (10 μM) increased the firing rate of VLPO LTS cells, reversed by flumazenil (5 μM), a GABAA benzodiazepine site antagonist. However, the firing rate of VLPO non-LTS cells was inhibited by ILTG (10 μM), also reversed by flumazenil (5 μM). No differences were detected regarding resting membrane potential (RMP) amplitude, spike threshold, afterhyperpolarization (AHP) amplitude, or action potential duration (APD50) after ILTG (10 μM) perfusion in VLPO LTS cells. RMP amplitude was more hyperpolarized and spike threshold was higher after ILTG (10 μM) application in VLPO non-LTS cells. In addition, ILTG significantly reduced the frequency of miniature inhibitory postsynaptic currents (mIPSCs) in VLPO LTS cells. ILTG significantly increased the amplitude of mIPSCs in VLPO non-LTS cells. Conclusions:This study revealed that ILTG suppresses presynaptic GABA release on VLPO LTS cells, thereby increasing their excitability. ILTG enhances postsynaptic GABAA receptor function on VLPO non-LTS cells, thereby decreasing their excitability. These results suggest that ILTG may produce hypnotic effects by modulating the GABAergic synaptic transmission properties of these two cell types.

The hypothalamic supramammillary nucleus (SuM) plays a crucial role in controlling wakefulness, but the downstream target regions participating in this control process remain unknown. Here, using circuit-specific fiber photometry and single-neuron electrophysiology together with electroencephalogram, electromyogram and behavioral recordings, we find that approximately half of SuM neurons that project to the medial septum (MS) are wake-active. Optogenetic stimulation of axonal terminals of SuM-MS projection induces a rapid and reliable transition to wakefulness from non-rapid-eye movement or rapid-eye movement sleep, and chemogenetic activation of SuM MS projecting neurons significantly increases wakefulness time and prolongs latency to sleep. Consistently, chemogenetically inhibiting these neurons significantly reduces wakefulness time and latency to sleep. Therefore, these results identify the MS as a functional downstream target of SuM and provide evidence for the modulation of wakefulness by this hypothalamic-septal projection.

Background and objectives Dexmedetomidine (DEX) is widely used in clinical sedation which has little effect on cardiopulmonary inhibition, however the mechanism remains to be elucidated. The basal forebrain (BF) is a key nucleus that controls sleep-wake cycle. The horizontal limbs of diagonal bundle (HDB) is one subregions of the BF. The purpose of this study was to examine whether the possible mechanism of DEX is through the α2 adrenergic receptor of BF (HDB). Methods In this study, we investigated the effects of DEX on the BF (HDB) by using whole cell patch clamp recordings. The threshold stimulus intensity, the inter-spike-intervals (ISIs) and the frequency of action potential firing in the BF (HDB) neurons were recorded by application of DEX (2 µM) and co-application of a α 2 adrenergic receptor antagonist phentolamine (PHEN) (10 µM). Results DEX (2 µM) increased the threshold stimulus intensity, inhibited the frequency of action potential firing and enlarged the inter-spike-interval (ISI) in the BF (HDB) neurons. These effects were reversed by co-application of PHEN (10 µM). Conclusion Taken together, our findings revealed DEX decreased the discharge activity of BF (HDB) neuron via α 2 adrenergic receptors.

目的 研究Toll样受体7(TLR7)在SD大鼠三叉神经疼痛中的作用,并初步探讨TLR7在疼痛过程中通过激活核因子-κB(NF-κB)通路介导相关炎症因子的作用机制.方法 采用眶下神经缩窄术(ION-CCI)建立大鼠三叉神经痛(TN)模型.通过实时荧光定量聚合酶链反应(qRT-PCR)及免疫印迹法(Western blot)检测三叉神经节(TG)内TLR7表达变化.经灌胃给药方式向ION-CCI大鼠给予TLR7抑制剂羟基氯喹(HCQ),检测抑制后TG内TLR7及其下游信号通路NF-κB亚基p65、p-p65及炎症因子TNF-α、IL-1β表达的变化.结果 结扎眶下神经所致的三叉神经损伤后,大鼠TG内的TLR7表达增加(P<0.05).在给予TLR7抑制剂后,大鼠TG内TLR7和TNF-α、IL-1β表达降低,并且p65核内移位和磷酸化减弱,NF-κB 信号通路激活被抑制,IONCCI诱导的雄性SD 大鼠的机械疼痛得到缓解(P<0.05).结论 TG 内TLR7通过激活初级感觉神经元中的NF-κB介导炎症因子TNF-α、IL-1β的表达参与调控神经病理性疼痛.

Background: Trigeminal neuralgia is a common chronic maxillofacial neuropathic pain disorder, and voltagegated sodium channels (VSGCs) are likely involved in its pathology. Prior studies report that pituitary adenylate cyclase-activating polypeptide (PACAP), a neuropeptide highly expressed in the trigeminal ganglion, may contribute to dorsal root ganglion neuron excitability by modulating the Nav1.7.Objective: We investigated whether PACAP can regulate Nav1.7 through the mitogen-activated protein kinase/ ERK kinase/extracellular-signal-regulated kinase (MEK/ERK) pathway in the trigeminal ganglion after chronic constriction injury of the infraorbital nerve (ION-CCI) in rats.Study design: Sprague-Dawley rats underwent ION-CCI, followed by intrathecal injection of PACAP 6-38 (PAC1 receptor antagonist) and PD98059 (MEK/ERK antagonist). Quantitative real-time PCR and western blot were used to quantify ATF3, PACAP, ERK, p-ERK, and Nav1.7 expression.Results: The mechanical pain threshold decreased from day 3 to day 21 after ION-CCI and reached the lowest testing value by day 14; however, it increased after PACAP 6-38 and PD98059 injections. Additionally, ION-CCI surgery increased ATF3, PACAP, and p-ERK expression in the rat trigeminal ganglion and decreased Nav1.7 and PAC1 receptor expression; however, there was no difference in ERK expression. PACAP 6-38 injection significantly decreased PACAP, p-ERK, and Nav1.7 expression and increased the PAC1 receptor expression, with no change in ERK expression. Moreover, PD98059 injection decreased PACAP, p-ERK, and Nav1.7 expression and increased the expression of PAC1 receptor.Conclusion: After ION-CCI, PACAP in the rat trigeminal ganglion can modulate Nav1.7 through the MEK/ERK pathway via the PAC1 receptor. Further, PACAP inhibition alleviates allodynia in ION-CCI rats.

The mechanism of light-induced spatial memory deficits, as well as whether rhythmic expression of the pituitary adenylyl cyclase-activating polypeptides (PACAP)-PAC1 pathway influenced by light is related to this process, remains unclear. Here, we aimed to investigate the role of the PACAP-PAC1 pathway in light-mediated spatial memory deficits. Animals were first housed under a T24 cycle (12 h light:12 h dark), and then light conditions were transformed to a T7 cycle (3.5 h light:3.5 h dark) for at least 4 weeks. The spatial memory function was assessed using the Morris water maze (MWM). In line with behavioral studies, rhythmic expression of the PAC1 receptor and glutamate receptors in the hippocampal CA1 region was assessed by western blotting, and electrophysiology experiments were performed to determine the influence of the PACAP-PAC1 pathway on neuronal excitability and synaptic signaling transmission. Spatial memory was deficient after mice were exposed to the T7 light cycle. Rhythmic expression of the PAC1 receptor was dramatically decreased, and the excitability of CA1 pyramidal cells was decreased in T7 cycle-housed mice. Compensation with PACAP1-38, a PAC1 receptor agonist, helped T7 cycle-housed mouse CA1 pyramidal cells recover neuronal excitability to normal levels, and cannulas injected with PACAP1-38 shortened the time to find the platform in MWM. Importantly, the T7 cycle decreased the frequency of AMPA receptor-mediated excitatory postsynaptic currents. In conclusion, the PACAP-PAC1 pathway is an important protective factor modulating light-induced spatial memory function deficits, affecting CA1 pyramidal cell excitability and excitatory synaptic signaling transmission.

To explore the correlation of tumor necrosis factor-α-induced protein 8-like protein 3 (TIPE3) expressions in colorectal cancer (CRC) with tumor-immune infiltration and patient prognosis. Formalin-fixed paraffin-embedded tumor samples from CRC patients (n = 110) were used in this study. Immunohistochemistry staining of TIPE3 and three prognostic immune biomarkers (CD8, CD20, and CD66b) was conducted in the tumor tissues and adjacent normal tissues. A Cox regression analysis of univariate and multivariate variables was performed to assess the correlation between TIPE3 and patient prognosis. We found that TIPE3 was mainly expressed in the cytoplasm, with a small amount in the nucleus. The expression of TIPE3 in tumor tissues is significantly higher than in adjacent normal tissues, and it is significantly correlated with the survival rate of patients in tumor tissues (p = 0.0038) and adjacent normal tissues (p<0.0001). Patients with a high TIPE3 expression had a lower survival rate, while patients with a low TIPE3 expression had a higher survival rate. Univariate regression analysis showed that the TIPE3 expression in tumor tissues (p = 0.007), the TIPE3 expression in adjacent normal tissues (p<0.001), the number of CD8+ T cells in tumor tissues (p = 0.020), the number of CD20+ B cells in tumor tissues (p = 0.023), the number of CD20+ B cells in adjacent normal tissues (p = 0.023), the number of CD66b+ neutrophils in tumor tissues (p = 0.005), the number of CD66b+ neutrophils in adjacent normal tissues (p<0.001), lymphatic metastasis (p = 0.010), TNM stage (p = 0.013), and tumor grade (p = 0.027) were significantly correlated with overall survival (OS). These prognostic factors were then subjected to multivariate regression analysis, and the results showed that the expression of TIPE3, the number of CD8+ T cells, and the number of CD66b+ neutrophils were prognostic factors affecting the OS rate of CRC patients. We found that the TIPE3 protein is upregulated in CRC cancer tissues and is correlated with survival rate.

AIM: To reveal the roles of CXCL13 and miR-186-5p in a rat model (adult male Sprague-Dawley rats, 7-8 weeks old, 180-200 g) of trigeminal neuralgia (TN) established via chronic constriction injury of the infraorbital nerve (ION-CCI).MATERIAL and METHODS: The results of behavioural tests and the expression levels of miR-186-5p and CXCL13 in the trigeminal ganglion (TG) were compared between the sham and ION-CCI groups, as well as the consequences of the miR-186-5p mimic and inhibitor.RESULTS: Compared with the sham-operated rats, ION-CCI rats displayed mechanical hypersensitivity in the von Frey hair test. Western blotting revealed the upregulation of CXCL13 and downregulation of miR-186-5p in the TG of ION-CCI rats relative to their expression in sham rats. Furthermore, an miR-186-5p mimic decreased CXCL13 protein levels and increased the mechanical withdrawal thresholds of ION-CCI rats. CXCL13 protein levels also increased after the injection of an miR-186-5p inhibitor. Finally, miR-186-5p was found to be expressed in the TG and was downregulated in ION-CCI rats compared to sham rats.CONCLUSION: miR-186-5p may negatively regulate CXCL13 to influence the occurrence and development of TN. Collectively, our findings shed new light on novel therapies for the treatment of TN.

Background: Circular RNA (circRNA) has an important influence on oral squamous cell carcinoma (OSCC) progression as competing endogenous RNAs (ceRNAs). However, the link between ceRNAs and the OSCC immune microenvironment is unknown. The research aimed to find circRNAs implicated in OSCC carcinogenesis and progression and build a circRNA-based ceRNA network to create a reliable OSCC risk prediction model.Methods: The expression profiles of circRNA in OSCC tumors and normal tissues were assessed through RNA sequencing. From the TCGA database, clinicopathological data and expression patterns of microRNAs (miRNAs) and mRNAs were obtained. A network of circRNA-miRNA-mRNA ceRNA was prepared according to these differentially expressed RNAs and was analyzed through functional enrichment. Subsequently, based on the mRNA in the ceRNA network, the influence of the model on prognosis was then evaluated using a risk prediction model. Finally, considering survival, tumor-infiltrating immune cells (TICs), clinicopathological features, immunosuppressive molecules, and chemotherapy efficacy were analyzed.Results: Eleven differentially expressed circRNAs were found in cancer tissues relative to healthy tissues. We established a network of circRNA-miRNA-mRNA ceRNA, and the ceRNA network includes 123 mRNAs, six miRNAs, and four circRNAs. By the assessment of Genomes pathway and Kyoto Encyclopedia of Genes, it is found that in the cellular senescence, PI3K-AKT and mTOR signaling pathway mRNAs were mainly enrichment. An immune-related signature was created utilizing seven immune-related genes in the ceRNA network after univariate and multivariate analysis. The receiver operating characteristic of the nomogram exhibited satisfactory accuracy and predictive potential. According to a Kaplan-Meier analysis, the high-risk group's survival rate was signally lower than the group with low-risk. In addition, risk models were linked to clinicopathological characteristics, TICs, immune checkpoints, and antitumor drug susceptibility.Conclusion: The profiles of circRNAs expression of OSCC tissues differ significantly from normal tissues. Our study established a circRNA-associated ceRNA network associated with OSCC and identified essential prognostic genes. Furthermore, our proposed immune-based signature aims to help research OSCC etiology, prognostic marker screening, and immune response evaluation.