ent-Herqueidiketal and epi-Peniciherqueinone Isolated from a Mushroom Derived Fungus Penicillium herquei YNJ-35.

A new polyaromatic metabolite, ent-herqueidiketal (1), and a new phenalenone derivative, epi-peniciherqueinone (2), along with twelve known compounds 3-14, were isolated from the fungus Penicillium herquei YNJ-35, a symbiotic fungus of Pulveroboletus brunneopunctatus collected from Nangunhe Nature Reserve, Yunnan Province, China. The structures of 1-14 and the absolute configurations of 1 and 2 were determined by their spectroscopic data or by their single-crystal X-ray diffraction analysis or optical rotation values. Compound 1 showed strong antibacterial activity against Staphylococcus aureus (ATCC 29213) with minimum inhibitory concentration (MIC) of 8 µg/mL. In the cytotoxicity assays, compound 1 showed weak inhibitory activity against breast cancer MCF-7 and mice microglial BV2 cells with half maximal inhibitory concentration (IC50 ) of 17.58 and 29.56 µM; compound 14 showed stronger cytotoxicity against BV2 and MCF-7 cells with IC50 values of 6.57 and 10.26 µM.

Ligustilide ameliorates cognitive impairment via AMPK/SIRT1 pathway in vascular dementia rat.

Vascular dementia (VaD) is the second cause of dementia after Alzheimer's disease. Ligustilide (LIG) is one of the main active ingredients of traditional Chinese medicines, such as Angelica. Studies have reported that LIG could protect against VaD. However, the mechanism is still confused. In this study, we employed a bilateral common carotid artery occlusion rat model to study. LIG (20 or 40 mg/kg/day) and Nimodipine (20 mg/kg) were orally administered to the VaD rats for four weeks. Morris water maze test showed that LIG effectively ameliorated learning and memory impairment in VaD rats. LIG obviously reduced neuronal oxidative stress damage and the level of homocysteine in the brain of VaD rats. Western blot results showed that pro-apoptotic protein Bax and cleaved caspase 3 increased and anti-apoptotic protein Bcl-2 decreased in the hippocampi of VaD rats. But after LIG treatment, these changes were reversed. Moreover, Nissl staining result showed that LIG could reduce neuronal degeneration in VaD rats. Furthermore, LIG enhanced the expressions of P-AMPK and Sirtuin1(SIRT1) in VaD rats. In conclusion, these studies indicated that LIG could ameliorate cognitive impairment in VaD rats, which might be related to AMPK/SIRT1 pathway activation.

Eleven-Residue Peptaibols Isolated from Trichoderma longibrachiatum Rifai DMG-3-1-1and Their Structure-Activity Relationship.

Total 23 eleven-residue peptaibols, including five reported ones (1-5) in our previous work, were isolated from the fungus Trichoderma longibrachiatum Rifai DMG-3-1-1, which was obtained from the mushroom Clitocybe nebularis (Batsch) P. Kumm. The structures of the 13 new peptaibols (6-10 and 12-19) were determined by their NMR and MALDI-MS/MS data, their absolute structures were further determined by Marfey's analyses and their ECD data. Careful comparison of the structures of 1-23 showed that only seven residues varied including the 2nd (Gln2 /Asn2 ), 3rd (Ile3 /Val3 ), 4th (Ile4 /Val4 ), 6th (Pro6 /Hyp6 ), 8th (Leu8 /Val8 ), 10th (Pro10 /Hyp10 ) and 11th (Leuol11 /Ileol11 /Valol11 ) residues. Comparison of the IC50 s against the three tested cell lines of 1-23 indicated that 2nd, 3rd and 4th amino acid residues affected their cytotoxicities powerfully. Compounds 2, 5, 9, 11, 21 and 22 showed moderate antibacterial activities against Staphylococcus aureus MRSA T144, which also showed stronger cytotoxicities against BV2 and MCF-7 cells.

Chemical constituents from the stems of Acanthopanax senticosus with their cytotoxic activities.

A new coumestan named 7,5'-dihydroxy-4'-(3''-hydroxy-3''-methyl-trans-isobut-1''-enyl) coumestan (1), together with five known compounds (2-6), was isolated from the EtOAc-soluble extract of the stems of Acanthopanax senticosus. Their structures were elucidated based on extensive spectroscopic analyses. All the isolates were evaluated for in vitro cytotoxic activities against four human cancer cells including HepG2, A549, HeLa and MCF-7. Among them, the new compound 1 was found to exhibit significant cytotoxic activity on HeLa cells with IC50 value of 6.5 µM.

Increasing efficiency of human mesenchymal stromal cell culture by optimization of microcarrier concentration and design of medium feed.

BACKGROUND AIMS: Large amounts of human mesenchymal stromal cells (MSCs) are needed for clinical cellular therapy. In a previous publication, we described a microcarrier-based process for expansion of MSCs. The present study optimized this process by selecting suitable basal media, microcarrier concentration and feeding regime to achieve higher cell yields and more efficient medium utilization. METHODS: MSCs were expanded in stirred cultures on Cytodex 3 microcarriers with media containing 10% fetal bovine serum. Process optimization was carried out in spinner flasks. A 2-L bioreactor with an automated feeding system was used to validate the optimized parameters explored in spinner flask cultures. RESULTS: Minimum essential medium-α-based medium supported faster MSC growth on microcarriers than did Dulbecco's modified Eagle's medium (doubling time, 31.6 ± 1.4 vs 42 ± 1.7 h) and shortened the process time. At microcarrier concentration of 8 mg/mL, a high cell concentration of 1.08 × 10(6) cells/mL with confluent cell concentration of 4.7 × 10(4)cells/cm(2) was achieved. Instead of 50% medium exchange every 2 days, we have designed a full medium feed that is based on glucose consumption rate. The optimal medium feed that consisted of 1.5 g/L glucose supported MSC growth to full confluency while achieving the low medium usage efficiency of 3.29 mL/10(6)cells. Finally, a controlled bioreactor with the optimized parameters achieved maximal confluent cell concentration with 16-fold expansion and a further improved medium usage efficiency of 1.68 mL/10(6)cells. CONCLUSIONS: We have optimized the microcarrier-based platform for expansion of MSCs that generated high cell yields in a more efficient and cost-effective manner. This study highlighted the critical parameters in the optimization of MSC production process.

Transfer of anterolateral thigh flaps in elderly oral cancer patients: complications in oral and maxillofacial reconstruction.

PURPOSE: Although a promising approach, the use of anterolateral thigh (ALT) free flaps has been limited in the reconstruction of oral and maxillofacial defects in elderly patients. The aim of this study was to estimate the frequency of postoperative complications and identify factors associated with complications. PATIENTS AND METHODS: The authors designed and implemented a retrospective study on the frequency of postoperative complications in elderly patients. They enrolled a sample composed of patients who underwent ALT free flap transfers for the repair of defects created during oral and maxillofacial cancer surgery from February 2002 to March 2013. The χ(2) test, t test, and multivariate regression model were used. RESULTS: A total of 1,100 patients were studied (859 men and 241 women). One hundred four patients (9.5%) were at least 70 years old (elderly group) at the time of surgery; the other 996 patients were younger than 70 years (younger group). The overall success rate of ALT free flap transfer was 97.2% (97.0% in the younger group, 99.0% in the elderly group; P > .05). The overall complication rate was 27.5% (27.2% in the younger group, 29.8% in the elderly group; P = .572). Multivariate analysis showed that operation time, American Society of Anesthesiologists class, and comorbidity were independent risk factors for postoperative complications in elderly patients. CONCLUSIONS: Oral and maxillofacial reconstruction using ALT free flaps in elderly patients can achieve outcomes similar to those obtained in younger patients. Limiting the operation time is important for improving surgical outcomes.

Application of Thinned Anterolateral Thigh Flap for the Reconstruction of Head and Neck Defects.

PURPOSE: To evaluate the feasibility and reconstructive efficacy of the thinned anterolateral thigh (ALT) flap for the reconstruction of head and neck defects. PATIENTS AND METHODS: A retrospective review was performed of 43 patients who had undergone reconstruction of head and neck defects with a thinned ALT flap from January 2009 through December 2013 at the Second Xiangya Hospital. The methods for flap thinning and defect reconstruction and reconstructive efficacy are reported. RESULTS: The flaps were 5 cm × 7 cm to 9 cm × 14 cm, and all were harvested from a single cutaneous perforator. Postoperatively, 40 flaps survived completely and 3 flaps experienced partial necrosis. Venous compromise occurred in 2 cases, both of which were salvaged after operative exploration. Of the 43 donor sites, 41 were closed directly and resulted in only linear scars, and 2 were closed using full-thickness skin grafts because the defect was larger. All the patients were followed for approximately 6 to 36 months, and they were satisfied with the esthetic and functional results of the donor and recipient sites after reconstruction. CONCLUSION: With the high success rate of flap transplantation, satisfactory functional and esthetic results, and lower complication rates at the donor and recipient sites, the use of thinned ALT flaps can be a good choice for the reconstruction of head and neck defects in obese patients.

A dual-mode sensing platform coupling two-signal ratiometric and colorimetric methods for detecting Au3+ based on surface state-regulated carbon nanodot.

The considerable risk posed by Au3+ residues to the environment and human health has sparked interest in researching Au3+ monitoring techniques. The detection results in the usual ratio mode are more reliable. In this work, we develop a dual-mode strategy based on reducing carbon dots coupling with two-signal ratiometric and colorimetric methods for high-sensitivity, good-selectivity, and wide-range detection of Au3+. Cyan carbon dots (C-CDs) were synthesized by a simple and efficient one-step hydrothermal method. The C-CDs with rich amino group used m-phenylenediamine as carbon source, which made it have the potential as a reducing agent. After the addition of Au3+, Au3+ was reduced to Au0, generating stable gold nanoparticles (AuNPs). The fluorescence signal (F490) of C-CDs decreased. At the same time, the large size of AuNPs enhances the second-order scattering signal (S770) and produces the UV-visible absorption peak of AuNPs. Therefore, the dual-mode sensing strategy combining S770/F490 ratiometric and colorimetric detection of Au3+ is realized with high accuracy and sensitivity. Au3+ was determined in real samples and a good recovery was obtained. The dual-mode method has good performance and practicality, so it shows great potential for environment testing in a simple and reliable way.

Sesquiterpenoid derivatives isolated from a liquid culture of Hypholoma capnoides 819.

Nine previously undescribed sesquiterpenoids, named as capnoidones A-G (1-6 and 8) and capnoidols A and B (7 and 9), along with three known sesquiterpenoids, fascicularones A, B, and G (12, 11 and 10), were isolated from the fermentation products of the mushroom Hypholoma capnoides 819 (Strophariace). The structures of these compounds were determined through MS and NMR experiments along with electronic circular dichroism analysis. Optical rotation calculations and X-ray diffraction experiments were also conducted for confirmation of the structures. Compounds 1 and 4 displayed mild cytotoxicity towards BV2 microglial cells in mice, while compound 4 exhibited mild cytotoxicity against breast cancer MCF-7 cells. However, none of the compounds demonstrated antibacterial activity against Staphylococcus aureus or Escherichia coli.

Anesthetic management of a patient with preoperative R-on-T phenomenon undergoing laparoscopic-assisted sigmoid colon resection: A case report.

BACKGROUND: The R-on-T phenomenon is a malignant arrhythmia associated with potentially catastrophic consequences. It may initiate ventricular tachycardia or ventricular fibrillation, which can result in syncope or sudden cardiac death. This manifestation poses a great challenge for anesthesiologists. However, it is rarely encountered in the perioperative setting. CASE SUMMARY: We herein present a case in which the R-on-T phenomenon was incidentally revealed by 24-h Holter monitoring in a patient diagnosed with sigmoid colon cancer. Careful evaluation and treatment with mexiletine were carried out preoperatively under consultation with a cardiovascular specialist, and surgery was uneventfully performed under general anesthesia after thorough preparation. CONCLUSION: Physicians should be vigilant about this infrequent but potentially fatal arrhythmia. Our experience suggests that the anesthetic process can be greatly optimized with careful preparation.

Taohong siwu decoction ameliorates cognitive dysfunction through SIRT6/ER stress pathway in Alzheimer's disease.

ETHNOPHARMACOLOGICAL RELEVANCE: A growing number of people suffer from Alzheimer's disease (AD), but there is currently no effective treatment yet. Taohong Siwu Decoction (TSD) has been proved to take strong neuropharmacological activity on dementia, but the effect and mechanism of TSD against AD are still elusive. AIM OF STUDY: To investigate whether TSD could be effective in ameliorating cognitive deficits through SIRT6/ER stress pathway. MATERIALS AND METHODS: Herein, the APP/PS1 mice, an AD model, and HT-22 cell lines were utilized. Different dosages of TSD (4.25, 8.50 and 17.00 g/kg/d) were administered to the mice for 10 weeks by gavage. Following the behavioral tests, oxidative stress levels were measured using malondialdehyde (MDA) and superoxide dismutase (SOD) kits. Nissl staining and Western blot analyses were used to detect the neuronal function. Then, immunofluorescence and Western blot analysis were applied to evaluate silent information regulator 6 (SIRT6) and ER Stress related protein levels in APP/PS1 mice and HT-22 cells. RESULTS: Behavioral tests revealed that APP/PS1 mice administered with TSD orally took more time in the target quadrant, crossed more times in the target quadrant, had a higher recognition coefficient, and spent more time in the central region. In addition, TSD could ameliorate oxidative stress and inhibit neuronal apoptosis in APP/PS1 mice. Furthermore, TSD could up-regulate the SIRT6 protein expression and inhibit ER sensing proteins expressions, such as p-PERK and ATF6, in APP/PS1 mice and Aß1-42-treated HT22 cells. CONCLUSION: According to the abovementioned findings, TSD could alleviate cognitive dysfunction in AD by modulating the SIRT6/ER stress pathway.

Hand Hygiene among Anesthesiologists and Microorganisms Contamination in Anesthesia Environments: A Single-Center Observational Study.

Objective: To investigate the baseline levels of microorganisms' growth on the hands of anesthesiologists and in the anesthesia environment at a cancer hospital. Methods: This study performed in nine operating rooms and among 25 anesthesiologists at a cancer hospital. Sampling of the hands of anesthesiologists and the anesthesia environment was performed at a ready-to-use operating room before patient contact began and after decontamination. Results: Microorganisms' growth results showed that 20% (5/25) of anesthesiologists' hands carried microorganisms (> 10 CFU/cm 2) before patient contact began. Female anesthesiologists performed hand hygiene better than did their male counterparts, with fewer CFUs ( P = 0.0069) and fewer species ( P = 0.0202). Our study also found that 55.6% (5/9) of ready-to-use operating rooms carried microorganisms (> 5 CFU/cm 2). Microorganisms regrowth began quickly (1 hour) after disinfection, and increased gradually over time, reaching the threshold at 4 hours after disinfection. Staphylococcus aureus was isolated from the hands of 20% (5/25) of anesthesiologists and 33.3% (3/9) of operating rooms. Conclusion: Our study indicates that male anesthesiologists need to pay more attention to the standard operating procedures and effect evaluation of hand hygiene, daily cleaning rate of the operating room may be insufficient, and we would suggest that there should be a repeat cleaning every four hours.

[Suppressive effect of propofol on low-voltage-activated calcium currents in rat hippocampal neurons].

OBJECTIVE: To investigate the effect of propofol on the low-voltage-activated calcium currents [ICa(LVA)] in rat hippocampal neurons. METHODS: Hippocampal neurons were prepared from Wistar rats and cultured. ICa(LVA) was recorded using whole cell patch clamp technique. Different concentrations of propofol were added to the culture. The effect of propofol on ICa(LVA) was evaluated. RESULTS: ICa(LVA) was inhibited by propofol in a concentration-dependent manner. Propofol 3 µmol /L had no effect on ICa(LVA). Propofol 10, 30, 100 and 300 µmol/L reduced peak ICa(LVA) by (12.6 ± 4.1)%, (29.2 ± 5.7)%, (36.6 ± 5.3)%, (31.6 ± 2.6)% respectively with a mean IC50 of 16.8 µmol/L and Hill coefficient of 0.15. The V1/2 of activation curve was shifted from (-10 ± 1 )mV to (-11 ± 2 )mV. K was 12 ± 1 and 8 ± 1. The V1/2 of inactivation curve was shifted from (-25 ± 1) mV to (-25 ± 5) mV. K was 15 ± 1 and 16 ± 3. CONCLUSION: Propofol produces significant inhibition of ICa (LVA) calcium currents in the central neurons which may partly explain the anesthefic action of propofol on the central nervous system.

Immunopathological Roles of Neutrophils in Virus Infection and COVID-19.

ABSTRACT: Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has been spread around the world and is currently affecting global public health. Clinical evidence indicates that the elevated number of peripheral neutrophils and higher ratio of neutrophils-to-lymphocytes are correlated with severe outcomes in COVID-19 patients, suggesting the possible immunopathological role of neutrophils during SARS-CoV-2 infection. As an abundant innate immune cell type, neutrophils are well known for their contributions to antimicrobial defense. However, their dysfunction is also associated with different inflammatory signatures during the pathogenesis of infection. Herein, in this mini-review, we summarize the recent progress on the potential role of neutrophils during COVID-19-associated inflammatory responses. In particular, we highlight the interactions between neutrophils and viruses as well as the relationship of neutrophils with cytokine storm and thrombosis in COVID-19 patients. Lastly, we discuss the importance of neutrophils as potential therapeutic targets for COVID-19.

[Suppression effect of ketamine on transient outward potassium currents in rathippocampal neurons].

OBJECTIVE: To investigate the effect of ketamine on the transient outward potassium currents [IA] using whole-cell patch clamp technique. METHODS: Pyramidal neurons were enzymatically isolated from Wistar rat hippocampus. The effect of ketamine on the [IA] was assessed using whole-cell patch clamp technique. Different concentrations of ketamine were added and potassium currents were measured. RESULTS: [IA] was inhibited by ketamine in a concentration-dependent manner. 10 micromol/L ketamine had no effect on [IA]. The four concentrations of ketamine (30, 100, 300, 1000 micromol/L) reduced peak [IA] currents by (11+/-2)%, (22+/-3)%, (45+/-5)%, (53+/-5)% respectively, with a mean IC50 of (130+/-24) micromol/L and Hill coefficient of 1.19+/-0.56. The V1/2 of activation curve was shifted from (-8.70+/-0.13) mV to (-11.2+/-2.10) mV (n=8, P<0.05). The V1/2 of inactivation curve was shifted from (-75.53+/-7.98) mV to (-91.94+/-11.85) mV (n=8, P<0.05). CONCLUSION: Ketamine produces significant inhibition of potassium currents in the central neurons, which may least partly explain at the action of ketamine.

[Effect of ketamine on high-voltage-activated calcium currents in rat hippocampal neurons].

OBJECTIVE: To investigate the effect of ketamine on the high-voltage-activated calcium currents (ICa(HVA)) in rat hippocampal neurons. METHODS: Neurons were cultured from Wistar rat hippocampus. ICa(HVA) was recorded using whole-cell patch clamp technique. After application with ketamine at different concentrations (10, 30, 100, 300, and 1000 µmol/L), the effect of ketamine on ICa(HVA) was evaluated. RESULTS: ICa(HVA) was inhibited by ketamine in a concentration-dependent manner. Ketamine at 10 µmol/L showed no effect on ICa(HVA). Four concentrations of ketamine (30, 100, 300,and 1000 µmol/L) reduced the peak ICa(HVA) currents by (17.5 ∓ 4.5)%, (25.5 ∓ 6.9)%, (38.5 ∓ 4.1)%, and (42.3 ∓ 4.6)% respectively,with a mean half maximal inhibitory concentration of 68.2 µmol/L and Hill coefficient of 0.47. The maximal activation membrane potential was shifted to (5.3 ∓ 0.8) from (5.4 ∓ 0.9). The half maximal activation membrane potential of inactivation curve was shifted from(-26.7 ∓ 3.9) mV to(-32.8 ∓ 4.2) mV. CONCLUSION: Ketamine can remarkably inhibit calcium currents in the central neurons,which may explain at least partly the action of ketamine on central nervous system.

Factors limiting the recruitment of Quercus schottkyana, a dominant evergreen oak in SW China.

Quercus schottkyana is a dominant species of oak in the Asian evergreen broad-leaved forests in southwestern China but seedlings are uncommon and recruitment is rare. Annual acorn production by Q. schottkyana is variable and the acorns are exposed to a series of mortality risks. Understanding the factors that limit recruitment of the oak requires knowledge of the oak's life cycle from acorn production to germination and seedling establishment. In this study, we first tested the effects of acorn density on establishment of seedlings by placing batches of acorns at different densities throughout the study area. Second, we tested the effects of herbivores on seedling survival by erecting fences around both natural and transplanted seedling populations. Our results show that even though the rate of seedling establishment increases as acorn density increases (for 32-8000 acorns∙m-2), survival rates of seedlings in the field were generally low (0-0.6%). We show that seedling recruitment of Q. schottkyana is mainly limited to the acorn stage where 88% of the acorns died from the combined effects of desiccation and predation by weevils (Curculio) and bark beetles (Coccotrypes sp.). Herbivory results in the death of some seedlings and consequently also affects the recruitment of seedlings of Q. schottkyana.

A case report of invasive candidiasis and fungal osteomyelitis mimicking oropharyngeal carcinoma recurrence in an immunocompetent patient following transoral robotic surgery.

INTRODUCTION: Transoral robotic surgery(TORS) for the excision of oropharyngeal tumour is increasingly considered for its lower rates of postoperative complications and better functional outcomes. However, we report a case of invasive candidiasis in an immunocompetent patient with previous radiation therapy years ago who underwent TORS recently. CASE PRESENTATION: A 55year old Chinese female with previous radiation therapy for nasopharyngeal carcinoma diagnosed in 1986 underwent wide excision of a newly diagnosed oropharyngeal carcinoma with TORS. From 4 to 8 weeks post-op, an enlarging exophytic lesion at the surgical excision site was noted. Initial biopsy did not show malignancy or fungal organisms. A repeat biopsy under anaesthesia was done as CT imaging showed suspicion of either infective or tumour recurrence. The patient was treated with long term antifungals as final culture results showed invasive candidiasis. The surgical site showed no further lesions 6 months post-op. DISCUSSION: Previous radiation therapy to the oropharyngeal area predisposes a patient to colonization of fungal organisms. Despite TORS showing lower rates of infection, patients who have had previous radiotherapy may be more susceptible to infections as the dissection involves down to the pre-vertebral fascia. Hence, it is appropriate for such patients to have microbiological and fungal cultures performed for a lesion such as this and be treated according to the results of the culture. CONCLUSION: This case serves as a reminder of the risk of invasive candidiasis requiring early investigation and treatment in an immunocompetent patient with prior history of both radiation therapy and recent TORS.

Reconstruction design before tumour resection: A new concept of through-and-through cheek defect reconstruction.

OBJECTIVE: To explore a new method of reconstruction of through-and-through cheek defects and to evaluate this method's efficacy and patient prognosis. MATERIALS AND METHODS: This retrospective study included 70 patients who underwent reconstruction of through-and-through cheek defects. The surgical approach, design of facial skin incisions, selection and design of flaps, postoperative quality of life and prognosis of patients were recorded and reported. RESULTS: Postoperative quality of life gradually increased over time, and the mean scores of University of Washington Quality of Life (UW-QOL) Questionnaire was more than 80 at 1-year postoperatively. The appearance, oral competence, chewing, swallowing, speech and other oral functions were well recovered in about 90% of patients at 1-year postoperatively. CONCLUSION: This new idea of reconstruction before tumour resection, brings the effect of plastic and reconstructive surgery to a new height.

Inhibiting ROS-NF-κB-dependent autophagy enhanced brazilin-induced apoptosis in head and neck squamous cell carcinoma.

Autophagy modulation has been considered a potential therapeutic strategy for head and neck squamous cell carcinoma (HNSCC). A previous study confirmed that brazilin might possess significant anti-carcinogenic activity. However, whether brazilin induces autophagy and its roles in cell death in HNSCC are still unclear. In this study, we have shown that brazilin induced significant apoptosis in the Cal27 HNSCC cell line but not in oral keratinocyte cell line (OKC). In addition to showing apoptosis induction, we demonstrated the brazilin-induced autophagic response in the Cal27 cells, as evidenced by the formation of GFP-LC3 puncta, and also showed the upregulation of LC3-II and Beclin-1. Moreover, pharmacologically or genetically blocking autophagy enhanced the brazilin-induced apoptosis, indicating the cytoprotective role of autophagy in brazilin-treated Cal27 cells. Moreover, brazilin activated nuclear factor kappa B (NF-κB p65) nuclear translocation and increased NF-κB p65 reporter activity, which contributed to the upregulation of autophagy-related genes, including LC3-II and Beclin-1. Importantly, we found that brazilin triggered reactive oxygen species (ROS) generation in Cal27 cells. Furthermore, N-acetyl-cysteine (NAC), a ROS scavenger, abrogated the effects of brazilin on the NF-κB p65-dependent autophagy. Taken together, our results demonstrated that brazilin increased the NF-κB p65-dependent autophagy through the promotion of ROS signalling pathways in HNSCC. These data also suggest that a strategy of blocking ROS-NF-κB p65-dependent autophagy to enhance the activity of brazilin warrants further attention for the treatment of HNSCC.