It has been assumed that olfactory sensitivity is relatively consistent in different populations worldwide. Emerging, yet fragmented evidence lends credit to the hypothesis that olfactory sensitivity may be geographically diverse. To gain deeper insight regarding the interplay between geographical, demographic, and health factors in the context of olfactory sensitivity, we conducted a multicenter study comprising data from 1046 participants inhabiting 19 locations around the world. Our results revealed that location accounted for 17–20% of the variance in chemosensory sensitivity. Demographic and psychological factors related to working memory and depressive symptoms also contribute to explaining sensitivity to odors, accounting for 1.6–2.9% of the variance in chemosensory sensitivity. Thus, we conclude that inhabitants of different geographical regions may present different sensitivities to chemical stimuli. We discuss the factors that could potentially be included in future investigations to pinpoint even more precisely what factors determine differences in chemosensory sensitivity around the globe.

BACKGROUND:Alzheimer's disease (AD) is a global health event with progressive cognitive decline affecting millions of elderly people worldwide. Emerging evidence indicates that sevoflurane may be associated with AD progression. However, the underlying mechanism remains poorly understood. Herein, we explored the potential role of sevoflurane exposure in cognitive ability and the possible mechanisms of action in the early stage of Tg4510 (P301L tau) transgenic mice. METHODS:Using the novel object recognition test and Y-maze test, we assessed the change in cognitive abilities in 3-month-old male WT mice and Tg4510 mice with or without 3% sevoflurane exposure. By histological analysis and western blot, we determined the effect of sevoflurane exposure on tau pathology, synaptic function, and neuroinflammatory response. TMT-labeled proteomics technique coupled with bioinformatics analysis was performed to identify the potential key pathways or proteins involved in sevoflurane-induced cognitive deterioration. RESULTS:Behavioral test showed that sevoflurane exposure exacerbated cognitive impairment of young Tg4510 mice. Pathologically, immunofluorescence demonstrated that sevoflurane exposure increased tau phosphorylation, synaptic defects, and neuroinflammatory response, which were further supported by the result of immunoblotting in Tg4510 mice exposed to sevoflurane. Proteomic analysis revealed an obvious decrease in autophagy-related proteins including Sort1, Vps28, and Atg3 in the hippocampus of sevoflurane-exposed Tg4510 mice compared to the nonexposed group. As an upstream signaling of autophagy, the AKT/mTOR pathway was found to be inhibited in sevoflurane-exposed Tg4510 mice. Moreover, our data also validated an inhibition of autophagy signaling associated with sevoflurane exposure in the context of tau pathology, as indicated by the upregulated expression of p62 and the downregulated expression of Sort1, Vps28, and Atg3. CONCLUSION:These findings suggest that autophagy signaling appears to be a promising target for intervention in sevoflurane-induced cognitive impairment in the early tau pathology, which lays the foundation for further study of the underlying mechanisms.

Gelsemium elegans (G. elegans) is an extremely poisonous plant that is widely distributed in southern China and southeastern Asia. G. elegans poisoning events occur frequently in southern China, and are therefore an urgent public health problem requiring multidisciplinary action. However, the toxic components and toxicological mechanisms remain unclear. Here, we describe a systematic investigation on the toxic components of G. elegans, resulting in the isolation and identification of 120 alkaloids. Based on acute toxicity screening, the structure-toxicity relationship of Gelsemium alkaloids was proposed for the first time. Moreover, gelsedine- and humantenine-type alkaloids were detected in the clinical blood sample, and were confirmed to be causative in the poisoning. The most toxic compound, gelsenicine (1), had selective inhibitory effects toward ventral respiratory group (VRG) neurons in the medulla, which is the main brain region controlling respiration in the central nervous system. Gelsenicine (1) strongly inhibited the firing of action potentials in VRG neurons through its ability to stimulate GABAA receptors, the main receptors involved in inhibitory neurotransmission. Application of GABAA receptor antagonists successively reversed action potential firing in gelsenicine (1)-treated VRG neurons. Importantly, the GABAA receptor antagonists securinine and flumazenil significantly increased the survival of poisoned animals. Our findings provide insight into the components and mechanisms of G. elegans toxicity, and should assist the development of effective emergency treatments for G. elegans poisoning.

Sevoflurane is an inhalational anesthetic widely used in clinical settings. Accumulating evidence has shown that sevoflurane exposure may impair cognitive function, potentially contributing to Alzheimer's disease (AD)-related changes. However, the underlying mechanism remains poorly understood. In the present study, 4-month-old 5xFAD mice were used to investigate the effect of sevoflurane exposure on cognitive decline by Y-maze test and novel object recognition test. We found that sevoflurane exposure promoted the appearance of cognitive impairment of 5xFAD mice, accompanied with the deterioration of Aβ accumulation, synaptic defects, and neuroinflammation. Additionally, sevoflurane was also found to aggravate mitochondrial fission of 5xFAD mice, as indicated by the further upregulated expression of p-Drp1S616. Moreover, sevoflurane significantly increased mitochondrial damage and dysfunction of AD models both in vitro and in vivo experiments. Seahorse XF analysis further indicated that sevoflurane exposure facilitated a metabolic shift from oxidative phosphorylation to glycolysis. Further rescue experiments revealed that a key mechanism underlying sevoflurane-induced cognitive impairment was the excessive mitochondrial fission, as supported by the result that the mitochondrial fission inhibitor Mdivi-1 counteracted the sevoflurane-mediated deteriorative effects in 5xFAD mice. These findings provided evidence for a new mechanism of sevoflurane exposure accelerating AD-related cognitive decline.

Objective The activation state of microglia is known to occupy a central position in the pathophysiological process of cerebral inflammation. Autophagy is a catabolic process responsible for maintaining cellular homeostasis. In recent years, autophagy has been demonstrated to play an important role in neuroinflammation. Resolvin D1 (RvD1) is a promising therapeutic mediator that has been shown to exert substantial anti-inflammatory and proresolving activities. However, whether RvD1-mediated resolution of inflammation in microglia is related to autophagy regulation needs further investigation. The present study aimed to explore the effect of RvD1 on microglial autophagy and its corresponding pathways. Methods Mouse microglial cells (BV-2) were cultured, treated with RvD1, and examined by Western blotting, confocal immunofluorescence microscopy, transmission electron microscopy, and flow cytometry. Results RvD1 promoted autophagy in both BV-2 cells and mouse primary microglia by favoring the maturation of autophagosomes and their fusion with lysosomes. Importantly, RvD1 had no significant effect on the activation of mammalian target of rapamycin (mTOR) signaling. Furthermore, RvD1-induced mTOR-independent autophagy was confirmed by observing reduced cytoplasmic calcium levels and suppressed calcium/calmodulin-dependent protein kinase II (CaMK II) activation. Moreover, by downregulating ATG5, the increased phagocytic activity induced by RvD1 was demonstrated to be tightly controlled by ATG5-dependent autophagy. Conclusion The present work identified a previously unreported mechanism responsible for the role of RvD1 in microglial autophagy, highlighting its therapeutic potential against neuroinflammation.

It has been proven that neuroinflammation triggered by microglial activation is the pathogenesis of depression associated with sepsis. An endogenous lipid mediator known as resolvin D1 (RvD1) is known to have anti-inflammatory effects in a sepsis model. However, it remains unknown if the effects of RvD1 on inflammatory responses are regulated by microglial autophagy. The current study investigated the role of RvD1-induced microglial autophagy in neuroinflammation. The findings showed that RvD1 reverses LPS-induced autophagy inhibition in microglia. RvD1 treatment significantly inhibits inflammatory responses by preventing NF-κB nuclear translocation and microglial M1 phenotypic transition. RvD1 exhibits an attenuation of neurotoxicity in both in vivo and in vitro models of sepsis. Following RvD1 injection, depressive-like behaviors in SAE mice were significantly improved. Notably, the aforesaid effects of RvD1 were eliminated by 3-MA, demonstrating that microglial autophagy was modulated. In conclusion, our findings shed new light on the involvement of microglial autophagy in SAE and emphasize the potential benefits of RvD1 as a promising therapeutic agent in the treatment of depression.

目的 构建水通道3(AQP-3)/血凝素(HA)融合标签载体,使用原子力显微镜(AFM)观察AQP-3在细胞膜的分布.方法 采用重叠聚合酶链反应(PCR)法将AQP-3/HA标签基因克隆到AQP-3的DNA序列中,构建真核表达载体,挑取单克隆菌落PCR初步鉴定后进行测序鉴定;质粒转染细胞后,采用免疫荧光标记HA标签蛋白,激光共聚焦显微镜下观察荧光标记并拍照;胶体金标记转染成功的细胞上HA标签,AFM观察胶体金颗粒从而分析AQP-3蛋白分布.结果 将HA标签蛋白插入AQP-3第二个胞外段,即T149-Y150之间,成功构建了融合表达载体pIRES2-AQP3/HA-EGF;共聚焦显微镜观察到转染成功的细胞发绿色荧光,AQP-3/HA融合蛋白呈蓝色荧光,其主要分布于细胞膜;AFM观察到细胞表面凹凸不平并伸出片状伪足和丝状伪足,AQP-3/HA在细胞膜表面分布有差异性,其在突起部位分布较多.结论 将HA标签插入AQP-3第二个胞外段,可减小对其功能影响,联合胶体金和AFM识别HA标签蛋白,能够实现AQP-3在细胞膜的示踪标记.

Objective: Obeticholic acid (OCA), a potent farnesoid X receptor (FXR) agonist, is a promising drug for nonalcoholic fatty liver disease (NAFLD); however, it can cause liver injury, especially at high doses. Here, we investigated the role of FXR in the high-dose OCA-induced hepatoxicity in the condition of the NAFLD mouse model. Methods: Wild-type (WT) mice and FXR−/− mice were administered with over-dose OCA (0.40%) and high-dose OCA (0.16%), in a high-fat diet. RNA-seq on liver samples of mice fed with high-dose OCA was performed to dig out the prominent biological events contributing to hepatic fibrosis. Results: Over-dose OCA induced liver injury and shortened survival in WT mice, but not FXR−/− mice. High-dose OCA caused hepatic stellate cell activation and liver fibrosis in the presence of FXR. Furthermore, high-dose OCA induced cholesterol accumulation in livers via the upregulation of genes involved in cholesterol acquisition and downregulation of genes regulating cholesterol degradation in liver, leading to the production of interleukin -1β and an FXR-mediated inflammatory response. Conclusion: The high-dose OCA induced FXR-dependent hepatic injury via cholesterol accumulation and interleukin -1β pathway in the NAFLD mice.

Iodide (I−) is crucial to thyroid function, and its regulation in thyrocytes involves ion transporters and reactive oxygen species (ROS). However, the extent of 2Cl−/H+ exchanger (ClC-3) involvement in the iodide (I−) efflux from thyrocytes remains unclear. Therefore, we examined the effects of ClC-3 on I− efflux. ClC-3 expression was found to significantly alter the serum TT3 and TT4 concentrations in mice. We further found that excess I− stimulation affected ClC-3 expression, distribution, and I− efflux in FRTL-5 cells. Immunofluorescence analyses indicated that ClC-3 mainly accumulated in the cell membrane and co-localized with β-tubulins after 24 h of excess I− treatment, and that this process depended on ROS production. Thus, ClC-3 may be involved in I− efflux at the apical pole of thyrocytes via excess I−-induced ROS production and β-tubulin polymerization. Our results reveal novel insights into the role of ClC-3 in I− transport and thyroid function.

Objective: Obeticholic acid (OCA), a farnesoid X receptor (FXR) agonist, is believed to alleviate nonalcoholic fatty liver disease (NAFLD) by decreasing hepatic lipogenesis in an FXR-dependent manner. Here, we revealed a novel mechanism by which OCA improves NAFLD by affecting hepatic long-chain fatty acids (LCFAs) uptake. Methods: Stably transfected HEK-293 cells expressing fatty acid transport protein 5 (FATP5) were established to examine fatty acid uptake; FXR-/-, human (h) FATP5, and FXR-/-/hFATP5 mouse models were incorporated to explore the effects of OCA on FATP5 ex vivo and in vivo. Results: OCA inhibited hFATP5 (IC50 =0.07 mu M) more than murine (m) FATP5 (IC50 =1.04 mu M) as measured by LCFAs uptake in FATP5 expressing HEK-293. OCA also inhibited LCFA uptake in primary hepatocytes from hFATP5 mice, FXR-/-/hFATP5 mice more than that from FXR(-/-)mice, ex vivo. Moreover, OCA inhibited LCFAs uptake by livers in hFATP5 mice and FXR-/-/hFATP5 mice, but not in FXR-/- mice, in vivo. Long-term administration of 0.04% OCA markedly reduced hepatic triglyceride (TG) accumulation in hFATP5 mice and FXR-/-/ hFATP5 mice by 63% and 53%, respectively, but not in FXR-/- mice. Conclusions: OCA ameliorated high-fat diet-induced NAFLD independent of FXR by inhibiting hepatic hFATP5mediated LCFAs uptake. This suggests that the therapeutic effects of OCA on NAFLD in vivo are mediated by a novel, hFATP5 dependent mechanism.

Acetaldehyde (ACD), the first metabolite of ethanol, is implicated in several of ethanol's actions, including the reinforcing and aversive effects. The neuronal mechanisms underlying ACD's aversive effect, however, are poorly understood. The lateral habenula (LHb), a regulator of midbrain monoaminergic centers, is activated by negative valence events. Although the LHb has been linked to the aversive responses of several abused drugs, including ethanol, little is known about ACD. We, therefore, assessed ACD's action on LHb neurons in rats. The results showed that intraperitoneal injection of ACD increased cFos protein expression within the LHb and that intra-LHb infusion of ACD induced conditioned place aversion in male rats. Furthermore, electrophysiological recording in brain slices of male and female rats showed that bath application of ACD facilitated spontaneous firing and glutamatergic transmission. This effect of ACD was potentiated by an aldehyde dehydrogenase (ALDH) inhibitor, disulfiram (DS), but attenuated by the antagonists of dopamine (DA) receptor (DAR) subtype 1 (SCH23390) and subtype 2 (raclopride), and partly abolished by the pretreatment of DA or DA reuptake blocker (GBR12935; GBR). Moreover, application of ACD initiated a depolarizing inward current (IACD) and enhanced the hyperpolarizing-activated currents in LHb neurons. Bath application of Rp-cAMPs, a selective cAMP-PKA inhibitor, attenuated ACD-induced potentiation of EPSCs and IACD Finally, bath application of ZD7288, a selective blocker of hyperpolarization-activated cyclic nucleotide-gated channels, attenuated ACD-induced potentiation of firing, EPSCs, and IACD These results show that ACD exerts its aversive property by exciting LHb neurons via multiple cellular mechanisms, and new treatments targeting the LHb may be beneficial for alcoholism.SIGNIFICANCE STATEMENT Acetaldehyde (ACD) has been considered aversive peripherally and rewarding centrally. However, whether ACD has a central aversive property is unclear. Here, we report that ACD excites the lateral habenula (LHb), a brain region associated with aversion and negative valence, through multiple cellular and molecular mechanisms. Intra-LHb ACD produces significant conditioned place aversion. These results suggest that ACD's actions on the LHb neurons might contribute to its central aversive property and new treatments targeting the LHb may be beneficial for alcoholism.

Persistent hypotonic and inflammatory conditions in the joint cavity can lead to the loss of cartilage matrix and cell death, which are the important mechanisms of osteoarthritis (OA) onset. Previous studies have confirmed that the existence of a hypotonic environment is a red flag for inflammation, as hypotonic environment induces the opening of the chloride channel of the cell and promotes chloride ion efflux, which prompts the cell volume to increase. Chloride channels play an important role in the regulation of mineralization and chondrocyte death. Here, we reported that OA chondrocytes showed a significant increase of cell death rate and the imbalance of cartilage matrix catabolism. We found that the distribution of skeleton protein F-actin was disordered. In addition, the volume-sensitive chloride current of OA chondrocytes decreased significantly with the increase of the expression levels of inflammation-related proteins caspase-1, caspase-3, and NLRP3. Moreover, interleukin-1β (IL-1β) showed a potential to activate the chloride current of normal chondrocytes. These results indicate that IL-1β-induced chloride channel opening in chondrocytes may be closely related to the occurrence of OA. This chloride channel opening process may therefore be a potential target for the treatment of OA.

Invasion and metastasis of breast cancer cells is an important cause of death in breast cancer patients. In the tumor microenvironment, M2 polarization of macrophages can promote the invasion and metastasis of tumor cells. OVOL2 is an evolutionarily conserved transcription regulator, but its effect in macrophages has not been described previously. The aim of this study was to investigate the effects of OVOL2 on macrophage polarity and the role of these effects in the tumor metastasis. We found that overexpression of OVOL2 in macrophages significantly inhibited M2 polarization and thus inhibits breast cancer metastasis. We propose a novel mechanism in which OVOL2 inhibits M2 polarization of macrophages and thus reduces their ability to induce invasion and metastasis of breast cancer. By shedding new light on the regulation of metastasis in cancers, our study provides a new strategy for the targeted therapy of cancer.

Estradiol regulates thyroid function, and chloride channels are involved in the regulation of thyroid function. However, little is known about the role of chloride channels in the regulation of thyroid functions by estrogen. In this study, the effects of estrogen on chloride channel activities in human thyroid Nthy-ori3-1 cells were therefore investigated using the whole cell patch-clamp technique. The results showed that the extracellular application of 17β-estradiol (E2) activated Cl- currents, which reversed at a potential close to Cl- equilibrium potential and showed remarkable outward rectification and an anion permeability of I- > Br- > Cl- > gluconate. The Cl- currents were inhibited by the chloride channel blockers, NPPB and tamoxifen. Quantitative Real-time PCR results demonstrated that ClC-3 expression was highest in ClC family member in Nthy-ori3-1 cells. The down-regulation of ClC-3 expression by ClC-3 siRNA inhibited E2-induced Cl- current. The Cl- current was blocked by the estrogen receptor antagonist, ICI 182780 (fulvestrant). Estrogen receptor alpha (ERα) and not estrogen receptor beta was the protein expressed in Nthy-ori3-1 cells, and the knockdown of ERα expression with ERα siRNA abolished E2-induced Cl- currents. Estradiol can promote the accumulation of ClC-3 in cell membrane. ERα and ClC-3 proteins were partially co-localized in the cell membrane of Nthy-ori3-1 cells after estrogen exposure. The results suggest that estrogen activates chloride channels via ERα in normal human thyroid cells, and ClC-3 proteins play a pivotal role in the activation of E2-induced Cl- current.

目的:探讨ClC-3氯通道在顺铂诱导的鼻咽癌细胞凋亡中的作用及机制.方法:将不同浓度的顺铂作用于人低分化鼻咽癌细胞(CNE-2Z细胞);MTT法检测不同浓度的顺铂处理24h和48h后细胞活力;采用ClC-3-siRNA下调ClC-3的表达,Annexin V-FITC/PI双染法检测细胞凋亡,多功能微孔板检测仪和流式细胞术检测细胞不同状态下的ROS水平.结果:(1)顺铂呈时间和浓度依赖性抑制CNE-2Z细胞生长,氯通道阻断剂DIDS显著抑制顺铂引起的细胞凋亡(P＜0.01);(2)顺铂促进CNE-2Z细胞ClC-3表达,下调ClC-3蛋白表达后,顺铂诱导的细胞凋亡率下降(P＜0.05);(3)顺铂促进CNE-2Z细胞ROS产生,用抗氧化剂抑制细胞产生ROS后,顺铂诱导的ClC-3蛋白表达和凋亡被抑制,而下调ClC-3蛋白表达对ROS水平影响不大.结论:顺铂可通过提升CNE-2Z细胞ROS水平,上调氯通道ClC-3蛋白水平,从而诱导细胞凋亡.

Background: Endometriosis is a progressive and benign disease characterized by the presence of endometrial glands and stroma tissue outside of the uterine cavity. Though endometriosis is a benign disease, it has the characteristics of malignant tumour growth. Abnormal expression of T-cadherin is involved in the occurrence and progression of many tumours. We aimed to investigate whether T-cadherin promotes the migration and invasion of endometriosis cells through the PI3K/AKT/mTOR signaling pathway. Methods: Ectopic and eutopic endometrial samples from 62 female patients with endometriosis and endometrial samples from 51 female patients without endometriosis were collected. The immortalized endometrial stromal cell line hEM15A was cultured. Real-time RT-PCR, immunohistochemistry and Western blot were used to detect the expression of T-cadherin, phospho-PI3K/Akt/mTOR and matrix metalloproteinase 2 (MMP-2). Transfection technology was employed to upregulate T-cadherin expression. The migration and invasion abilities of hEM15A cells were measured by the transwell assay with uncoated or Matrigel-coated membranes. Results: The mRNA and protein expression of T-cadherin was significantly decresed in the ectopic tissues of the patients with endometriosis, while the mRNA and protein expression in the eutopic endometrial tissues of the same patients did not significantly differ from that in the patients without endometriosis. The migration and invasion ability and phospho-PI3K/Akt/mTOR and MMP-2 expression levels were decreased in hEM15A cells with high T-cadherin expression compared with the corresponding parameters in the normal control group. However, everolimus and BEZ235 inhibited cell migration and invasion in cells with low T-cadherin expression, and weakened overexpression of T‑cadherin significantly attenuated MMP-2 protein expression. Conclusion: Loss of T-cadherin promotes cell migration and invasion in endometriosis via the PI3K/AKT/mTOR signalling pathway.

Extracellular acidification, playing a promoting role in the process of acute pancreatitis, has been reported to activate Cl- channels in several types of cells. However, whether extracellular acidification aggravates acute pancreatitis via activating Cl- channels remains unclear. Here, we investigated the effects of extracellular acidification on Cl- channels in rat pancreatic acinar AR42J cells using whole-cell patch-clamp recordings. We found that extracellular acidification induced a moderately outward-rectified Cl- current, with a selectivity sequence of I- > Br- >= Cl- > gluconate(-), while intracellular acidification failed to induce the currents. The acid-sensitive currents were inhibited by Cl- channel blockers, 4,4'-Diisothiocyanatostilbene-2,2'-disulfonic acid disodium salt hydrate and 5-Nitro 2 (3-phenylpropylamino) benzoic acid. After ClC-3 was silenced by ClC-3 shRNA, the acid-sensitive Cl- currents were attenuated significantly, indicating that ClC-3 plays a vital role in the induction of acid-sensitive Cl- currents. Extracellular acid elevated the intracellular level of reactive oxygen species (ROS) significantly, prior to inducing Cl- currents. When ROS production was scavenged, the acid-sensitive Cl(-)currents were abolished. Whereas, the level of acid-induced ROS was unaffected with silence of ClC-3. Our findings above demonstrate that extracellular acidification induces a Cl- current in pancreatic acinar cells via promoting ROS generation, implying an underlying mechanism that extra-cellular acidification might aggravate acute pancreatitis through Cl- channels. (C) 2020 Elsevier Inc. All rights reserved.

Nutrient deficiency develops frequently in nasopharyngeal carcinoma cell (CNE-2Z) due to the characteristics of aggregation and uncontrolled proliferation. Therefore, starvation can induce autophagy in these cells. Chloride channel 3 (ClC-3), a member of the chloride channel family, is involved in various biological processes. However, whether ClC-3 plays an important role in starvation-induced autophagy is unclear. In this study, Earle's balanced salt solution (EBSS) was used to induce autophagy in CNE-2Z cells. We found that autophagy and the chloride current induced by EBSS were inhibited by chloride channel blockers. ClC-3 knockdown inhibited the degradation of LC3-II and P62. Furthermore, when reactive oxygen species (ROS) generation was suppressed by antioxidant N-acetyl-l-cysteine (L-NAC) pretreatment, EBSS-induced autophagy was inhibited, and the chloride current was unable to be activated. Nevertheless, ClC-3 knockdown had little effect on ROS levels, indicating that ROS acted upstream of ClC-3 and that both ROS and ClC-3 participated in EBSS-induced autophagy regulation in CNE-2Z.

Propofol infusion syndrome (PRIS) is an uncommon life-threatening complication observed most often in patients receiving high-dose propofol. High-dose propofol treatment with a prolonged duration can damage the immune system. However, the associated molecular mechanisms remain unclear. An increasing number of clinical and experimental observations have demonstrated that tissue-resident macrophages play a critical role in immune regulation during anaesthesia and procedural sedation. Since the inflammatory response is essential for mediating propofol-induced cell death and proinflammatory reactions, we hypothesised that propofol overdose induces macrophage pyroptosis through inflammasomes. Using primary cultured bone marrow-derived macrophages, murine macrophage cell lines (RAW264.7, RAW-asc and J774) and a mouse model, we investigated the role of NLRP3 inflammasome activation and secondary pyroptosis in propofol-induced cell death. We found that high-dose propofol strongly cleaved caspase-1 but not caspase-11 and biosynthesis of downstream interleukin (IL)-1β and IL-18. Inhibition of caspase-1 activity blocks IL-1β production. Moreover, NLRP3 deletion moderately suppressed cleaved caspase-1 as well as the proportion of pyroptosis, while levels of AIM2 were increased, triggering a compensatory pathway to pyroptosis in NLRP3-/- macrophages. Here, we show that propofol-induced mitochondrial reactive oxygen species (ROS) can trigger NLRP3 inflammasome activation. Furthermore, apoptosis-associated speck-like protein (ASC) was found to mediate NLRP3 and AIM2 signalling and contribute to propofol-induced macrophage pyroptosis. In addition, our work shows that propofol-induced apoptotic initiator caspase (caspase-9) subsequently cleaved effector caspases (caspase-3 and 7), indicating that both apoptotic and pyroptotic cellular death pathways are activated after propofol exposure. Our studies suggest, for the first time, that propofol-induced pyroptosis might be restricted to macrophage through an NLRP3/ASC/caspase-1 pathway, which provides potential targets for limiting adverse reactions during propofol application. These findings demonstrate that propofol overdose can trigger cell death through caspase-1 activation and offer new insights into the use of anaesthetic drugs.

Although the spontaneous chloride currents (SCC) have been well studied in the normal cells, its properties and roles in neoplasms cells are still unknown. Here, we found that the SCC was manifested in the poorly differentiated human nasopharyngeal carcinoma CNE-2Z cells, with some differences such as lower occurrence and bigger current density than those of the volume-activated chloride currents (VACC). NPPB, a chloride channel blocker, inhibited the SCC much stronger than the VACC. Down-regulation of chloride channel -3 (ClC-3), a volume and mechanically dependent ion channel, could significantly decrease the VACC, but not in SCC. The occurrence, latency, and mean density of the SCC were much lower in the normal nasopharyngeal NP69-SV40T cells than those in CNE-2Z cells. Our results demonstrated that the spontaneous electrical reactivity of neoplasm cells is higher than that of normal cells, which probably relates to their high physiological activity of neoplasm cells. Significance of the study Spontaneous chloride currents (SCC) are well known in excitable tissues and regulate a variety of physiological and pathophysiological processes. During our researching on the volume-activated chloride currents (VACC) in human nasopharyngeal carcinoma CNE-2Z cells, SCC could be also observed with different properties from VACC. Meanwhile, the occurrence, latency, and mean density of the SCC were much higher in CNE-2Z cells than those in normal nasopharyngeal NP69-SV40T cells. Our results revealed the expression and characteristics of SCC in carcinoma cells and provided a preliminary experimental basis for further exploring the function of SCC in tumour cell biology.