Low-Temperature Adaptive Single-Atom Iron Nanozymes against Viruses in the Cold Chain.

Outbreaks of viral infectious diseases, such as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and influenza A virus (IAV), pose a great threat to human health. Viral spread is accelerated worldwide by the development of cold chain logistics; Therefore, an effective antiviral approach is required. In this study, it is aimed to develop a distinct antiviral strategy using nanozymes with low-temperature adaptability, suitable for cold chain logistics. Phosphorus (P) atoms are added to the remote counter position of Fe-N-C center to prepare FeN4P2-single-atom nanozymes (SAzymes), exhibiting lipid oxidase (OXD)-like activity at cold chain temperatures (-20, and 4 °C). This feature enables FeN4P2-SAzymes to disrupt multiple enveloped viruses (human, swine, and avian coronaviruses, and H1-H11 subtypes of IAV) by catalyzing lipid peroxidation of the viral lipid envelope. Under the simulated conditions of cold chain logistics, FeN4P2-SAzymes are successfully applied as antiviral coatings on outer packaging and personal protective equipment; Therefore, FeN4P2-SAzymes with low-temperature adaptability and broad-spectrum antiviral properties may serve as key materials for developing specific antiviral approaches to interrupt viral transmission through the cold chain.

Discovery of 4-amino-1,6-dihydro-7H-pyrrolo[2,3-d]pyridazin-7-one derivatives as potential receptor-interacting serine/threonine-protein kinase 1 (RIPK1) inhibitors.

Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) is an important regulatory factor in the necroptosis signaling pathway, and is considered an attractive therapeutic target for treating multiple inflammatory diseases. Herein, we describe the design, synthesis, and structure-activity relationships of 4-amino-1,6-dihydro-7H-pyrrolo [2,3-d]pyridazin-7-one derivatives as RIPK1 inhibitors. Among them, 13c showed favorable RIPK1 kinase inhibition activity with an IC50 value of 59.8 nM, and high RIPK1 binding affinity compared with other regulatory kinases of necroptosis (RIPK1 Kd = 3.5 nM, RIPK3 Kd = 1700 nM, and MLKL Kd > 30,000 nM). 13c efficiently blocked TNFα-induced necroptosis in both human and murine cells (EC50 = 1.06-4.58 nM), and inhibited TSZ-induced phosphorylation of the RIPK1/RIPK3/MLKL pathway. In liver microsomal assay studies, the clearance rate and half-life of 13c were 18.40 mL/min/g and 75.33 min, respectively. 13c displayed acceptable pharmacokinetic characteristics, with oral bioavailability of 59.55%. In TNFα-induced systemic inflammatory response syndrome, pretreatment with 13c could effectively protect mice from loss of body temperature and death. Overall, these compounds are promising candidates for future optimization studies.

Supragastric lesser sac: an insidious site for surgical exploration during the debulking surgery in advanced ovarian cancer.

OBJECTIVE: Metastases in the supragastric lesser sac (SGLS) are not only occult but are also barriers to complete resection of ovarian cancer. We describe a cohort of patients with SGLS disease undergoing debulking surgery. METHODS: We identified all patients who underwent evaluation and eventual resection of SGLS disease as part of cytoreductive surgery for stage IIIC-IVB high-grade epithelial ovarian cancer at our institution from January 2018 to August 2022. RESULTS: Thirty-three of 286 patients (11.5%) underwent resection of SGLS disease. Metastases in the SGLS were identified by preoperative imaging in 4 of 33 patients (12.1%). The median peritoneal cancer index score was 22 (range, 9-33). Through surgical exploration, metastases were frequently seen in the right diaphragm (100%), hepatorenal recess (97%), lesser omentum (81.8%), left diaphragm (78.8%), supracolic omentum (75.8%), anterior transverse mesocolon (72.7%), splenic hilum (63.6%), ligamentum teres hepatis (60.6%), and gallbladder fossa (51.5%). The lesser omentum was normal in 6 of 33 (18.2%) patients, despite metastases within the SGLS. A total of 54.5% of patients underwent complex surgery (surgical complexity scores; median, 8; range, 3-14). Complete resections were achieved in 19 (57.6%) patients. No complications were related to the resection of SGLS disease. The median length of progression-free survival was 24.8 months (95% confidence interval=16.6-32.9). CONCLUSION: Metastases to the SGLS are not uncommon in advanced ovarian cancer, particularly those with widely disseminated disease. Disease in this recess is rarely identified by preoperative imaging and deserves systematic surgical exploration to attain complete cytoreduction.

Effects of solid-state fermentation with Bacillus subtilis LK-1 on the volatile profile, catechins composition and antioxidant activity of dark teas.

In this study, the solid-state fermentation (SSF) of dark tea was carried out using Bacillus subtilis LK-1, which was isolated from Fu brick tea (FBT). The effects of SSF with B. subtilis on volatile organic compounds (VOCs), non-volatile metabolites, and antioxidant activities of dark tea was investigated. A total of 45 VOCs were identified, primarily consisting of ketones (18), hydrocarbons (8), aldehydes (7), and alcohols (6). Following fermentation, the content of key odor active substances such as linalool, ß-ionone, and 3,5-octadiene-2-one significantly increased, resulting in an enhanced floral and fruity aroma of dark tea. Furthermore, new flavor substances like geranyl isovalerate and decanal were produced during SSF, enriching the aroma profile of dark tea. Non-ester catechins demonstrated a drastic increase, while ester catechins remarkably decreased after SSF. Furthermore, SSF led to a slight decrease in the total polyphenols content and antioxidant activity of dark tea. There is a close relationship between VOCs and the main non-volatile metabolites during SSF. Overall, this study highlighted the great impact of SSF with B. subtilis on the metabolites of dark tea and provided valuable insights into the role of bacteria in shaping the metabolite profile of FBT.

The Way to Malignant Transformation: Can Epigenetic Alterations Be Used to Diagnose Early-Stage Head and Neck Cancer?

Identifying and treating tumors early is the key to secondary prevention in cancer control. At present, prevention of oral cancer is still challenging because the molecular drivers responsible for malignant transformation of the 11 clinically defined oral potentially malignant disorders are still unknown. In this review, we focused on studies that elucidate the epigenetic alterations demarcating malignant and nonmalignant epigenomes and prioritized findings from clinical samples. Head and neck included, the genomes of many cancer types are largely hypomethylated and accompanied by focal hypermethylation on certain specific regions. We revisited prior studies that demonstrated that sufficient uptake of folate, the primary dietary methyl donor, is associated with oral cancer reduction. As epigenetically driven phenotypic plasticity, a newly recognized hallmark of cancer, has been linked to tumor initiation, cell fate determination, and drug resistance, we discussed prior findings that might be associated with this hallmark, including gene clusters (11q13.3, 19q13.43, 20q11.2, 22q11-13) with great potential for oral cancer biomarkers, and successful examples in screening early-stage nasopharyngeal carcinoma. Although one-size-fits-all approaches have been shown to be ineffective in most cancer therapies, the rapid development of epigenome sequencing methods raises the possibility that this nonmutagenic approach may be an exception. Only time will tell.

Risk Factors of Postoperative Cerebrospinal Fluid Leak After Craniovertebral Junction Anomalies Surgery: A Case-Control Study.

OBJECTIVE: To identify potential risk factors for cerebrospinal fluid (CSF) leakage after craniovertebral junction (CVJ) anomaly surgery and to provide a reference for clinical practice. METHODS: Sixty-six patients who underwent elective CVJ anomaly surgery during a 6-year period (April 2013 to September 2019) were retrospectively included. Research data were collected from the patients' medical records and imaging systems. Patients were divided into CSF leak and no CSF leak groups. Univariate tests were performed to identify potential risk factors. For statistically significant variables in the univariate tests, a logistic regression test was used to identify independent risk factors for CSF leakage. RESULTS: The overall prevalence of CSF leakage was 13.64%. Univariate tests showed that a basion-dental interval (BDI) > 10 mm and occipitalized atlas had significant intergroup differences (p < 0.05). Multivariate analysis indicated that a BDI > 10 mm was an independent risk factor for CSF leakage, and patients with CVJ anomalies with a BDI > 10 mm were more likely to have postoperative CSF leaks (odds ratio, 14.67; 95% confidence interval, 1.48-30.88; p = 0.004). CONCLUSION: It is necessary to maintain vigilance during CVJ anomaly surgery in patients with a preoperative BDI > 10 mm to avoid postoperative CSF leaks.

Bidirectionally Regulating Viral and Cellular Ferroptosis with Metastable Iron Sulfide Against Influenza Virus.

Influenza virus with numerous subtypes and frequent variation limits the development of high-efficacy and broad-spectrum antiviral strategy. Here, a novel multi-antiviral metastable iron sulfides (mFeS) against various influenza A/B subtype viruses is developed. This work finds that mFeS induces high levels of lipid peroxidation and •OH free radicals in the conservative viral envelope, which depends on Fe2+ . This phenomenon, termed as a viral ferroptosis, results in the loss of viral infectibility and pathogenicity in vitro and in vivo, respectively. Furthermore, the decoction of mFeS (Dc(mFeS)) inhibits cellular ferroptosis-dependent intracellular viral replication by correcting the virus-induced reprogrammed sulfur metabolism, a conserved cellular metabolism. Notably, personal protective equipment (PPE) that is loaded with mFeS provides good antiviral protection. Aerosol administration of mFeS combined with the decoction (mFeS&Dc) has a potential therapeutic effect against H1N1 lethal infection in mice. Collectively, mFeS represents an antiviral alternative with broad-spectrum activity against intracellular and extracellular influenza virus.

PA-X protein of H9N2 subtype avian influenza virus suppresses the innate immunity of chicken bone marrow-derived dendritic cells.

H9N2 subtype avian influenza (AI) is an infectious disease associated with immunosuppression in poultry. Here, the regulation function of PA-X protein was determined on the host innate immune response of H9N2-infected chicken bone marrow-derived DCs (chBM-DCs). Based on 2 mutated viruses expressing PA-X protein (rTX) or deficient PA-X protein (rTX-FS), and the established culture system of chBM-DCs, results showed PA-X protein inhibited viral replication in chBM-DCs but not in non-immune chicken cells (DF-1). Moreover, PA-X protein downregulated the expression of phenotypic markers (CD40, CD86, and MHCII) and proinflammatory cytokine (IL-12 and IL-1ß) of chBM-DCs. The mixed lymphocyte reaction between chBM-DCs and chicken T cells showed PA-X protein significantly decreased H9N2-infected chBM-DCs to induce T cell proliferation, implying a suppression of the DC-induced downstream T cell response. Taken together, these findings indicated that PA-X protein is a key viral protein to help H9N2 subtype AIVs escape the innate immunity of chBM-DCs.

Who really benefits from intraperitoneal chemotherapy for advanced ovarian cancer? A treatment-free survival analysis of the AICE trial.

OBJECTIVE: To investigate whether peritoneal disease extent can predict the survival benefit of intraperitoneal/intravenous (IP/IV) chemotherapy in ovarian cancer. DESIGN: A treatment-free survival (TFS) analysis. SETTING: Five-centre trial. POPULATION: An extended follow-up of the Additional Intraperitoneal Cisplatin and Etoposide in ovarian cancer (AICE) trial (NCT01669226), with data cut-off on 27 August 2020. Patients were categorised into subgroups with high tumour burden (HTB) and low tumour burden (LTB). METHODS: Overall survival (OS) was divided into time on protocol treatment exposure (T), time free of subsequent treatment or death (TFS) and time after the first subsequent therapy (REL). TFS analyses and quality-adjusted OS were calculated by multiplying the mean time in each health state by its assigned utility: quality-adjusted OS = ut  × T + TFS + urel  × REL. MAIN OUTCOME MEASURES: The area under each Kaplan-Meier curve was estimated using the 96-month restricted mean time, with threshold utility analyses used to illustrate quality-adjusted OS comparisons. RESULTS: In the HTB subgroup, the restricted mean TFS was 33.9 months and 18.7 months in the IP/IV and IV groups, respectively (p = 0.005), with a significant quality-adjusted OS gain (13.2-16.0 months). In the LTB subgroup, IP/IV therapy yielded no survival benefit in either TFS (p = 0.268) or quality-adjusted OS (range: 1.4-6.3 months). CONCLUSIONS: Both TFS and quality-adjusted OS was longer across all utility weight values with IP/IV than with standard IV therapy in the HTB subgroup, whereas patients in the LTB subgroup did not benefit from the therapy. The tumour burden of ovarian cancer should be assessed before deciding on IP/IV versus IV treatment.

Effect of Amomum villosum essential oil as an additive on the chemical composition, fermentation quality, and bacterial community of paper mulberry silage.

Paper mulberry (Broussonetia papyrifera L., PM) is being used as a new type of animal protein feed to address the feed crisis. To investigate the effect of additives on the chemical composition, fermentation quality, and bacterial community of PM silage (at room temperature, 25°), paper mulberry was fermented with formic acid (FA), Amomum villosum essential oil (AVEO) and lactic acid bacteria (LAB) inoculant treatments. The results showed that fresh PM had a low water-soluble carbohydrate (WSC) content and large amounts of unclassified bacteria. Compared with the CK and LAB treatments, the FA and AVEO treatments significantly (P < 0.05) decreased the pH and increased the lactic acid content of PM silage after 60 days of ensiling. In the AVEO-treated silages the abundance of Lactococcus in the early stage of ensiling increased by 14.09%, the abundances of Levilactobacillus and Lentilactobacillus in the late stage of ensiling increased by 58.34 and 91.12%, respectively, and the abundance of Stenotrophomonas decreased by 94.71%, resulting in improved PM silage quality. These results confirmed that AVEO could potentially be developed as a new additive for improving the fermentation quality of silage.

PA-X protein assists H9N2 subtype avian influenza virus in escaping immune response of mucosal dendritic cells.

H9N2 subtype low pathogenicity avian influenza virus (AIV) poses a potential zoonotic risk. PA-X, a novel protein generated by PA gene ribosomal frameshift, is considered to be the virulence factor of H9N2 subtype AIVs. Our study found that rTX possessing PA-X protein enhanced the mammalian pathogenicity of H9N2 subtype AIVs compared with PA-X-deficient virus (rTX-FS). Furthermore, PA-X protein inhibited H9N2 subtype AIVs to infect dendritic cells (DCs), but not nonimmune cells (MDCK cells). Meanwhile, PA-X protein suppressed the phenotypic expression (CD80, CD86, CD40 and MHCII), early activation marker (CD69) and pro-inflammatory cytokines (IL-6 and TNF-α), whereas increased anti-inflammatory cytokine (IL-10) in DCs. After intranasally viral infection in mice, we found that PA-X protein of H9N2 subtype AIVs reduced CD11b+ and CD103+ subtype mucosal DCs recruitment to the nasal submucosa by inhibiting CCL20 expression. Moreover, PA-X protein abolished the migratory ability of CD11b+ and CD103+ DCs into draining cervical lymph nodes by down-regulating CCR7 expression. The rTX-infected DCs significantly impaired the allogeneic CD4+ T cell proliferation, suggesting PA-X protein suppressed the immune functions of DCs for hindering the downstream immune activation. These findings indicated that PA-X protein assisted H9N2 subtype AIVs in escaping immune response of mucosal DCs for enhancing the pathogenicity.

Effect of the enhanced recovery after surgery protocol on recovery after laparoscopic myomectomy: a systematic review and meta-analysis.

Background: Surgery is the recommended treatment for uterine leiomyoma but it still has issues like postoperative complications and slow recovery. The enhanced recovery after surgery (ERAS) protocol could probably reduce traumatic stress and promote the rapid postoperative recovery of patients, but there are controversies for the results of different studies. This meta-analysis was performed to resolve the controversies and provide evidence for the application of ERAS in gynecology. Methods: The PubMed, Embase, Ovid, CNKI (China), Wanfang Data (China), and Google Scholar databases were searched to recruit all studies on the application of ERAS in laparoscopic myomectomy up to November 2021. The inclusion criteria of studies was established according to the PICOS principles. the Cochrane RoB 2.0 and Newcastle-Ottawa Scale (NOS) scale were used to assess the bias of the studies, RevMan 5.3 software was used for meta-analysis. Results: Ten studies that met the criteria were finally included with 1,441 participants. Eight of them were randomized controlled trials (RCTs) and two were cohort studies, all of them were with low level of bias. Meta-analysis showed that ERAS protocol after laparoscopic myomectomy could significantly shorten the first time getting out of bed after surgery [mean difference (MD) =-4.85; 95% confidence interval (CI): (-7.35, -2.36); P=0.0001], the first defecation time after surgery [MD =-4.69; 95% CI: (-5.68, -3.69); P<0.00001], and the postoperative hospital stay [MD =-1.32, 95% CI: (-2.08, -0.56); P=0.0007]. It could also markedly reduce the patient readmission rate [odds ratio (OR) =0.42; 95% CI: (0.23, 0.76); P=0.004], and notably reduced the incidence of complications [OR =0.37; 95% CI: (0.22, 0.61); Z=3.82; P=0.0001]. Yet, the cost of the ERAS protocol was not significantly different from that of routine care [MD =-127.76, 95% CI: (-997.19, 741.66); P=0.77]. Discussion: The application of ERAS protocol after gynecological laparoscopic myomectomy can shorten the first defecation time, first time out of bed, hospital stay, and reduce the readmission rate as well as the incidence of postoperative complications, without additional costs. But still there was heterogeneity among the studies, the topic still deserved further exploration.

Comparison of the Fungal Community, Chemical Composition, Antioxidant Activity, and Taste Characteristics of Fu Brick Tea in Different Regions of China.

In this study, the fungal community structure, metabolites, antioxidant ability, and taste characteristics of five Fu brick tea (FBT) from different regions of China were determined and compared. A total of 69 operational taxonomic units (OTUs) were identified and assigned into 5 phyla and 27 genera, with Eurotium as the predominant genus in all samples. Hunan (HN) sample had the strongest fungal diversity and richness, followed by Guangxi (GX) sample, and Zhejiang (ZJ) sample had the lowest. GX sample had higher amounts of gallic acid (GA), total catechins, gallocatechin (GC), and epicatechin gallate (ECG) as well as antioxidant activity than the other samples. The levels of total phenolics, total flavonoids, epigallocatechin (EGC), catechin, epicatechin (EC), thearubigins (TRs), and theaflavins (TFs) were the highest in the ZJ sample. Guizhou (GZ) and Shaanxi (SX) samples contained the highest contents of epigallocatechin gallate (EGCG) and gallocatechin gallate (GCG), respectively. Total phenolics, GA, EC, CG, and TFs were positively associated with most of fungal genera. Total phenolic content (TPC), total flavonoid content (TFC), and most of catechins contributed to the antioxidant activities of FBT. HN sample had the strongest sourness and sweetness, ZJ sample had the strongest saltiness, SX sample had the strongest umami, and GZ sample had the strongest astringency, which was ascribed to the varied metabolites. This work reveals that FBT in different regions vary greatly in fungal community, metabolites, antioxidant activity, and taste characteristics, and provides new insight into the quality characteristics formation of FBT in different regions.

Bifunctional polypyrrole-based conductive paper towards simultaneous efficient solar-driven water evaporation and electrochemical energy storage.

The thriving solar-driven water evaporation (SDWE) technology is considered the ideal candidate for next-generation water treatment because of its high efficiency, environment-friendliness, and low cost. The irresistible trend of diversified energy demand presents multi-functional requirements for a successful SWDE. However, the current SDWE technology rarely breaks through this technical dilemma. Here, we have designed a bifunctional polypyrrole-based capacitor to achieve water purification and energy storage. The hydrophilicity of the filter paper and the high light absorptance of polypyrrole (96.18%) promote the generation of solar steam. The evaporation rate of the PPy-200 (Polypyrrole-200) filter paper reached 1.54 kg m-2 h-1 under 1 kW m-2. Interestingly, the symmetric supercapacitor assembled with PPy-based filter paper electrodes could simultaneously realize efficient evaporation (1.94 kg m-2 h-1) and electrochemical energy storage. As a single electrode, the PPy-200 filter paper exhibited ultra-high specific capacitance (4129.50 mF cm-2) and favorable cycling stability (71.16% after 4000 cycles). More importantly, the capacitance of PP-PPy-200 (Polyvinyl alcohol/Polyethylene glycol-Polypyrrole-200) increased to 2.55 times under one sun illumination. This work not only points out a direction for solar thermal utilization, but also provides new design inspiration for high-efficiency flexible electrochemical energy storage devices.

Oridonin alleviates hyperbilirubinemia through activating LXRα-UGT1A1 axis.

Hyperbilirubinemia is a serious hazard to human health due to its neurotoxicity and lethality. So far, successful therapy for hyperbilirubinemia with fewer side effects is still lacking. In this study, we aimed to clarify the effects of oridonin (Ori), an active diterpenoid extracted from Rabdosia rubescens, on hyperbilirubinemia and revealed the underlying molecular mechanism in vivo and in vitro. Here, we showed that liver X receptor alpha (LXRα) deletion eliminated the protective effect of Ori on phenylhydrazine hydrochloride-induced hyperbilirubinemia mice, indicating that LXRα acted as a key target for Ori treatment of hyperbilirubinemia. Ori significantly increased the expression of LXRα and UDP-glucuronosyltransferase 1A1 (UGT1A1) in the liver of wild-type (WT) mice, which were lost in LXRα-/- mice. Ori or LXR agonist GW3965 also reduced lipopolysaccharide/D-galactosamine-induced hyperbilirubinemia via activating LXRα/UGT1A1 in WT mice. Liver UGT1A1 enzyme activity was elevated by Ori or GW3965 in WT mice. Further, Ori up-regulated LXRα gene expression, increased its nuclear translocation and stimulated UGT1A1 promoter activity in HepG2 cells. After silencing LXRα by siRNA, Ori-induced UGT1A1 expression was markedly reduced in HepG2 cells and primary mouse hepatocytes. Taken together, Ori stimulated the transcriptional activity of LXRα, resulting in the up-regulation of UGT1A1. Therefore, Ori or its analogs might have the potential to treat hyperbilirubinemia-related diseases through modulating LXRα-UGT1A1 signaling.

Superparamagnetic Iron Oxide Nanoparticles Protect Human Gingival Fibroblasts from Porphyromonas gingivalis Invasion and Inflammatory Stimulation.

INTRODUCTION: Modulating the inflammatory response of human gingival fibroblasts (hGFs) is important for the control of periodontal inflammation because it is a key event in the pathogenesis of periodontitis. Here, we aimed to determine whether polyglucose sorbitol carboxymethyl ether (PSC)-coated superparamagnetic iron oxide nanoparticles (SPIONs) protect hGFs against invasion and inflammatory stimulation by Porphyromonas gingivalis (P. gingivalis). METHODS: First, we determined the cytotoxicity and antimicrobial activity of PSC-SPIONs. Then, their effects on invasion of hGFs by P. gingivalis were evaluated by counting invading P. gingivalis, fluorescence staining, and transmission electron microscopy. The effect of PSC-SPIONs on inflammation in hGFs induced by P. gingivalis lipopolysaccharide was evaluated by measurement of reactive oxygen species (ROS), and enzyme-linked immunosorbent assays, quantitative reverse transcription-polymerase chain reaction, and Western blotting of key indicator molecules. The effects of dimercaptosuccinic acid (DMSA)-coated SPIONs and the free form of PSC alone were also tested and compared with those of PSC-SPIONs. RESULTS: PSC-SPIONs (25 µg/mL) are cytocompatible with hGFs and exhibit no antimicrobial effects on P. gingivalis. However, they inhibit invasion of hGFs by P. gingivalis at 15 µg/mL. They also decrease ROS production and inflammatory cytokine secretion by hGFs at 5, 15, and 25 µg/mL, by downregulating activation of the nuclear factor-kappa B signaling pathway. Furthermore, PSC alone does not inhibit inflammation, while DMSA-SPIONs do. This indicates that the nanosize effects of PSC-SPIONs, rather than their coating material, play the dominant role in their anti-inflammatory activity. CONCLUSION: PSC-SPIONs protect hGFs against P. gingivalis invasion and inflammatory stimulation. Thus, they have potential for clinical application in control of periodontal inflammation.

Geraniin inhibits cell growth and promoted autophagy-mediated cell death in the nasopharyngeal cancer C666-1 cells.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a rare malignant tumor developing from epithelial linings of nasopharynx, and 10-50 out of 100,000 NPC cases were recorded globally particularly in the Asian countries. METHODOLOGY: The cytotoxicity of geraniin against the NPC C666-1 cells were analyzed using MTT assay. The influences of geraniin on the C666-1 cell viability with the presence of ROS and apoptosis inhibitors were also studied. The expressions of PI3K, Akt, mTOR, and autophagic markers LC3, ATG7, P62/SQSTM1 expressions in the C666-1 cells were studied by western blotting analysis. The ROS production was assayed using DCFH-DA staining. The immunofluorescence assay was performed to detect the NF-κB and ß-catenin expressions in the C666-1 cells. RESULTS: The cell viability of C666-1 cells were appreciably prevented by the geraniin. The geraniin treatment also inhibited the C666-1 cell growth with the presence of apoptotic inhibitor Z-VAD-FMK. The geraniin-treatment effectively improved the ROS production and inhibited the NF-κB and ß-catenin expressions in the C666-1 cells. Geraniin appreciably modulated the PI3K/Akt/mTOR signaling axis and improved the autophagy-mediated cell death via improving the autophagic markers LC3 and ATG7 expressions in the C666-1 cells. CONCLUSION: In conclusion, our results proved that geraniin inhibits C666-1 cell growth and initiated autophagy-mediated cell death via modulating PI3K/Akt/mTOR cascade and improving LC3 and ATG7 expressions in the C666-1. Geraniin and it could be a hopeful and efficient candidate to treat the human NPC in the future.