Unconventional hcp/fcc Nickel Heteronanocrystal with Asymmetric Convex Sites Boosts Hydrogen Oxidation.

Developing non-platinum group metal catalysts for the sluggish hydrogen oxidation reaction (HOR) is critical for alkaline fuel cells. To date, Ni-based materials are the most promising candidates but still suffer from insufficient performance. Herein, we report an unconventional hcp/fcc Ni (u-hcp/fcc Ni) heteronanocrystal with multiple epitaxial hcp/fcc heterointerfaces and coherent twin boundaries, generating rugged surfaces with plenty of asymmetric convex sites. Systematic analyses discover that such convex sites enable the adsorption of *H in unusual bridge positions with weakened binding energy, circumventing the over-strong *H adsorption on traditional hollow positions, and simultaneously stabilizing interfacial *H2O. It thus synergistically optimizes the HOR thermodynamic process as well as reduces the kinetic barrier of the rate-determining Volmer step. Consequently, the developed u-hcp/fcc Ni exhibits the top-rank alkaline HOR activity with a mass activity of 40.6 mA mgNi-1 (6.3 times higher than fcc Ni control) together with superior stability and high CO-tolerance. These results provide a paradigm for designing high-performance catalysts by shifting the adsorption state of intermediates through configuring surface sites.

[Identification of chemical constituents in Simiao Yong'an Decoction based on UPLC-LTQ-Orbitrap-MS].

This study aims to identify the chemical constituents of Simiao Yong'an Decoction based on ultra-performance liquid chromatography coupled with linear quadrupole ion trap-orbitrap mass spectrometry(UPLC-LTQ-Orbitrap-MS). The elution was performed through a UPLC BEH C_(18) column(2.1 mm × 100 mm, 1.7 µm) with the mobile phase of water(containing 0.1% formic acid)-acetonitrile at a flow rate of 0.4 mL·min~(-1). LTQ-Orbitrap-MS with heat electrospray ion(HESI) source was employed to collect MS fragment information in the negative ion mode. A total of 72 compounds were identified based on reference substance comparison, fragmentation rules, accurate molecular weight, related reports and databases(MassBank and HMDB), including 30 iridoid glycosides, 9 organic acids, 15 flavonoids, 10 phenylpropanoids, 7 triterpenoids, and 1 saccharide. The method established in this study is comprehensive, rapid, and accurate, which can help summarize the fragmentation rules of constituents and provide reference for revealing the active constituents and pharmacodynamic mechanism of Simiao Yong'an Decoction.

Soft Electrochemical Actuators with a Two-Dimensional Conductive Metal-Organic Framework Nanowire Array.

Electrically activated soft actuators capable of large deformation are powerful and broadly applicable in multiple fields. However, designing soft actuators that can withstand a high strain, provide a large actuation displacement, and exhibit stable reversibility are still the main challenges toward their practical application. Here, for the first time, we report a two-dimensional (2D) conductive metal-organic framework (MOF) based electrochemical actuator, which consists of vertically oriented and hierarchical Ni-CAT NWAs/CNF electrodes through the use of a facile one-step in situ hydrothermal growth method. The soft actuator prepared in this study demonstrated improvements in actuation performance and benefits from both the intrinsically ordered porous architecture and efficient transfer pathways for fast ion and electron transport; furthermore, this actuator facilitated a considerably high diffusion rate and low interfacial resistance. In particular, the actuator demonstrated a rapid response (<19 s) at a 3 V DC input, large actuation displacement (12.1 mm), and a correspondingly high strain of 0.36% under a square-wave AC voltage of ±3 V. Specifically, the actuator achieved a broad-band frequency response (0.1-20 Hz) and long-term cyclability in air (10000 cycles) with a negligible degradation in actuation performance. Our work demonstrates new opportunities for bioinspired artificial actuators and overcomes current limitations in electrode materials for soft robotics and bionics.

Simultaneous determination of five alkaloids from Rauvolfia vomitoria in rat plasma by LC-MS/MS: Application to a comparative pharmacokinetic study in normal and type 2 diabetic rats.

Rauvolfia vomitoria is widely distributed in the tropical regions of Africa and Asia, and has been used in traditional folk medicine in China. Indole alkaloids were found to be major bioactive components, while the effects of diabetes mellitus on the pharmacokinetic parameters of the components have not been reflected in vivo. In this study, an efficient and sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous determination of five ingredients of R. vomitoria in rats. Detection was implemented in multiple-reaction-monitoring mode with an electrospray positive-ionization source. Validation parameters were all in accordance with the current criterion. The established method was effectively employed to compare the pharmacokinetic behaviors of five alkaloids (reserpine, yohimbine, ajmaline, ajmalicine, and serpentine) between normal and type 2 diabetic rats. The single-dose pharmacokinetic parameters of the five alkaloids were determined in normal and diabetic rats after oral administration of 100 and 200 mg/kg body weight. The results indicated that diabetes mellitus significantly altered the pharmacokinetic characteristics of yohimbine, ajmaline, and ajmalicine after oral administration in rats. This is an attempt to provide some evidence for clinicians that may serve as a guide for the use of antidiabetic medicine in clinical practice.

Matrix-Assisted DOSY for Analysis of Indole Alkaloid Mixtures.

Diffusion-ordered spectroscopy (DOSY) is a powerful tool for investigating mixtures and identifying peaks of chemical components. However, similar diffusion coefficients of the components, particularly for complex mixtures that contain crowded resonances, limit resolution and restrict application of the DOSY technique. In this paper, matrix-assisted DOSY were used to explore whether the diffusion resolution of a complex model involving indole alkaloid mixtures can be realized. Furthermore, we investigated the influence of different factors on the separation effect. The results showed that the changes in diffusion coefficient differences were achieved more obviously when using sodium dodecyl sulfate (SDS) micelles as the matrix. In addition, we also found that increasing the concentration of SDS can improve the resolution of the DOSY spectrum. Finally, after investigating the influence factors and NMR conditions, we demonstrated the applications of the SDS-assisted DOSY on analyzing the total alkaloid extract of Alstonia Mairei, and the virtual separation of mixtures was achieved.

Research Progress of NMR in Natural Product Quantification.

In the fields of medicine and health, traditional high-performance liquid chromatography or UV-visible spectrophotometry is generally used for substance quantification. However, over time, nuclear magnetic resonance spectroscopy (NMR) has gradually become more mature. Nuclear magnetic resonance spectroscopy has certain advantages in the quantitative analysis of substances, such as being nondestructive, having a high flux and short analysis time. Nuclear magnetic resonance spectroscopy has been included in the pharmacopoeiae of various countries. In this paper, the principle of nuclear magnetic resonance spectroscopy and the recent progress in the quantitative study of natural products by NMR are reviewed, and its application in the quantitative study of natural products is proposed. At the same time, the problems of using NMR alone to quantify natural products are summarized and corresponding suggestions are put forward.

4ß-Hydroxywithanolide E from Goldenberry (Whole Fruits of Physalis peruviana L.) as a Promising Agent against Chronic Obstructive Pulmonary Disease.

Environmental toxicant- and oxidant-induced [e.g., cigarette smoke (CS)] respiratory oxidative stress and inflammatory response play a vital role in the onset and progression of COPD. The nuclear factor erythroid 2-related factor 2 (Nrf2) represents an important mechanism for regulating intracellular oxidative stress and inflammatory response and is a promising target for developing agents against COPD. Herein, a bioactivity-guided purification of goldenberry (whole fruits of Physalis peruviana L.) led to the isolation of a novel and potent Nrf2 activator 4ß-hydroxywithanolide E (4ß-HWE). Our study indicated that (i) 4ß-HWE activated the Nrf2-mediated defensive response through interrupting Nrf2-Keap1 protein-protein interaction (PPI) via modification of Cys151 and Cys288 cysteine residues in Keap1 and accordingly suppressing the ubiquitination of Nrf2. (ii) 4ß-HWE enhanced intracellular antioxidant capacity and inhibited oxidative stress in normal human lung epithelial Beas-2B cells and wild-type AB zebrafish. (iii) 4ß-HWE blocked LPS-stimulated inflammatory response and inhibited LPS-stimulated NF-κB activation in RAW 264.7 murine macrophages. (iv) 4ß-HWE effectively suppressed oxidative stress and inflammatory response in a CS-induced mice model of pulmonary injury. Collectively, these results display the feasibility of using 4ß-HWE to prevent or alleviate the pathological progression of COPD and suggest that 4ß-HWE is a candidate or a leading molecule against COPD.

Chemical Constituents from Physalis Calyx seu Fructus and Their Inhibitory Effects against Oxidative Stress and Inflammatory Response.

Physalis Calyx seu Fructus, a traditional Chinese medicine consisting of the calyxes and fruits of Physalis alkekengi var. franchetii, has been used as therapy for inflammation-related respiratory diseases such as excessive phlegm, cough, sore throat, and pharyngitis for a long history in China. The aim of the present study was to investigate the chemical constituents of Physalis Calyx seu Fructus and identify the bioactive constituents responsible for its traditional application as therapy for inflammation-related diseases. In the present study, one new phenylpropanoid (1: ), two new steroids (17: and 18: ), together with 55 known constituents have been purified from the EtOH extract of Physalis Calyx seu Fructus. Among them, seven and twelve known constituents were isolated for the first time from Physalis Calyx seu Fructus and the genus Physalis, respectively. Fourteen constituents, including steroids [physalins (5:  - 9, 12:  - 14: , and 15: ) and ergostane (21: )], a sesquiterpenoid (35: ), alkaloids (36: and 37: ), and a flavonoid (44: ), showed inhibitory effects against oxidative stress. Ten constituents, including steroids (5, 6, 8, 13: , and 15: ), sesquiterpenoids (34: and 35: ), alkaloids (37: and 41: ), and a flavonoid (43: ), were found be potential anti-inflammatory constituents of this medicinal plant. The inhibition of oxidative stress and inflammatory response may be related to the regulation of Nrf2 and nuclear factor-κB pathways. The ethnomedical use of Physalis Calyx seu Fructus as a treatment for respiratory diseases might be attributed to the combined inhibitory effects of steroids, alkaloids, sesquiterpenoids, and flavonoids against oxidative stress and inflammatory response.

[Simultaneous rapid detection of eight neurotransmitters in rat tissues,blood and urine samples by UPLC-MS/MS].

Ultra performance liquid chromatography-tandem mass spectrometry( UPLC-MS/MS) was used to establish the simultaneous determination method of eight neurotransmitters in brain,liver,kidney,adrenal gland,serum and urine,including serotonin,5-hydroxyindole acetic acid,epinephrine,norepinephrine,dopamine,glutamic acid,γ-aminobutyric acid,and acetylcholine,and then investigate the distribution characteristics of neurotransmitters in rat tissues,blood and urine. Waters ACQUITY UPLC BEH C_(18) column( 2. 1 mm×100 mm,1. 7 µm) was used,with 0. 3% formic acid solution-acetonitrile as the mobile phase for gradient elution.Multiple reaction monitoring( MRM) scanning method under positive mode by atmospheric pressure electrospray ion source( ESI) was also performed to establish the detection method of neurotransmitters for methodological research. The plasma,urine and tissues of normal rats were pre-treated including homogenization,centrifuging,and protein removal,then the 2 µL supernatant was injected for analysis. The results showed that eight kinds of neurotransmitters could be accurately determined within 7 min,with linear correlation coefficients all greater than 0. 99. This method showed high accuracy and good precision,with specificity,stability,extraction recovery and matrix effects all complying with the biological sample analysis requirements. The most abundant transmitters in the brain,liver,kidney,kidney gland,blood and urine were γ-aminobutyric acid,glutamic acid,glutamic acid,adrenaline,glutamic acid and dopamine.The method is sensitive,rapid,precise,accurate and specific,and can be used for simultaneous quantitative analysis of eight neurotransmitters in biological samples. The investigation of the distribution ratio of transmitters in rats is of important significance to disease prevention and treatment.

Nanoporous Gold Embedded ZIF Composite for Enhanced Electrochemical Nitrogen Fixation.

The electrochemical nitrogen reduction reaction (NRR) offers an energy-saving and environmentally friendly approach to produce ammonia under ambient conditions. However, traditional catalysts have extremely poor NRR performances because of their low activity and the competitive hydrogen evolution reaction. The high catalytic activity of nanoporous gold (NPG) and the hydrophobicity and molecular concentrating effect of the zeolitic imidazolate framework-8 (ZIF-8) were incorporated in the NPG@ZIF-8 nanocomposite so that the ZIF-8 shell could weaken hydrogen evolution and retard reactant diffusion. A highest Faradaic efficiency of 44 % and an excellent rate of ammonia production of (28.7±0.9) µg h-1 cm-2 were achieved, which are superior to traditional gold nanoparticles and NPG. Moreover, the composite catalyst shows high electrochemical stability and selectivity (98 %). The superior NRR performance makes NPG@ZIF-8 one of the most promising water-based NRR electrocatalysts for ammonia production.

A Double-Walled Bimetal-Organic Framework for Antibiotics Sensing and Size-Selective Catalysis.

The first double-walled bimetal-organic framework [Cd4K4(TADA)4(H2O)12]·6DMF (1, TADA = 3,3'-((6-hydroxy-1,3,5-triazine-2,4-diyl)bis(azanediyl)) dibenzoate) has been successfully constructed. In virtue of its high porosity and abundant basic sites, MOF 1 can be served as a high-efficiently size-selective heterogeneous catalyst for Knovenagel condensation reaction. Remarkably, 1 also exhibits excellent luminescent detectability for metronidazole (MDZ) with the highest quenching constant ( Ksv) of 1.16 × 105 M-1. The results demonstrate that 1 can be served as a unique bifunctional platform for antibiotics sensing and size-selective catalysis.

In Vivo and In Vitro Inhibitory Effects of Icariin on Angiogenesis.

OBJECTIVE: To investigate the inhibitory effects of icariin(ICA),an active ingredient of Herb Epimedii,on angiogenesis. METHODS: The chick chorioallantoic membrane(CAM)assay was adopted to evaluate the effects of various doses of the ICA on the angiogenesis. The cell growth inhibitory effect of ICA on human umbilical vein endothelial cells(HUVEC)was measured by MTT assay. Cell cycle arrest and the induction of apoptosis were evaluated by flow cytometry. The effect of ICA on the migration of HUVEC cells was measured on Transwell model. RESULTS: ICA remarkably inhibited angiogenesis in CAM in a concentration-dependent manner. The proliferation of HUVEC cells was inhibited by ICA, and the effect was time-and concentration-dependent. ICA-treated HUVEC cells showed cell cycle arrest;180 µg/ml of ICA decreased the percentage of migrating HUVEC cells by 78.0%. CONCLUSION: ICA can effectively suppress angiogenesis;however,its in vivo inhibitory effect on angiogenesis warrants further investigations.

Diterpenoids from the Chinese liverwort Heteroscyphus tener and their antiproliferative effects.

Four new ent-labdane diterpenoids, heteroscyphins A-D (1-4), and four known diterpenoids (5-8) were isolated from the Chinese liverwort Heteroscyphus tener (Steph.) Schiffn. The absolute configuration of compound 1 was defined by single-crystal X-ray diffraction using Cu Kα radiation. Cytotoxicity tests revealed that compounds 3 and 5 exhibited modest activity against seven cancer cell lines. Compound 5 showed inhibitory effects on prostate cancer (PCa) cell proliferation but with less inhibition on non-neoplastic prostate epithelial cells. Compound 5 markedly caused cell growth arrest at the G0/G1 phase and induced cellular apoptosis through ROS-mediated DNA damage in PCa cells.

[Effect of RNA interference targeting Schistosoma japonicum aldose reductase gene].

OBJECTIVE: To investigate the effect of siRNA-induced RNA interference (RNAi) at both messenger RNA (mRNA) and protein level, targeting the Schistosoma japonicum aldose reductase (SjAR) gene. METHODS: Two siRNAs (siRNA-1 and siRNA-2) were designed based on SjAR gene sequence and commercially synthesized together with a negative control siRNA (siRNA-NC) showing no homology to any known S. japonicum gene sequences. Each female Kunming mice (4-6 weeks old) was infected with 800-1,000 S. japonicum cercariae and sacrificed on Day 13 postinfection for schistosomula collection. Every 120 +/- 10 worms were selected as one group and subjected to RNAi treatment based on siRNA electroporation or soaking. In terms of the electroporation method, 5 microg of siRNA-1, siRNA-2, siRNA-NC or equal volumes of DEPC water (blank control) were added to each cuvette separately and the schistosomula in the cuvettes subsequently underwent a square wave pulse of 125 V and 20 ms duration, followed by a 48 h culture. As for the soaking method, solutions of the three siRNAs were added to the culture media separately to a concentration of 200 nmol/L, with equal volumes of DEPC water added as a blank control. Fresh media were changed and siRNAs were supplemented on Day 3 amid the 5 days' soaking process. Three repeats were set up for each treatment. Worm RNA and total soluble proteins were extracted after the RNAi treatment by using the TRIzol method, and cDNA was synthesized by reverse transcription of the RNA. Quantitative real-time PCR (qRT-PCR) was performed next to determine the change of relative SjAR transcripts abundance after the RNAi treatment with SjGAPDH as an internal reference. Western blotting was performed to measure the expression changes of SjAR at protein level, after degradation of the SjAR transcripts by RNAi treatment. RESULTS: qRT-PCR result showed that the relative expression level of SjAR transctripts was (90.6 +/- 16.2)%, (84.2 +/- 7.3)%, and (105.9 +/- 10.3)%, respectively, following the electroporation of siRNA-1, siRNA-2 or siRNA-NC as well as the 48 h culture, none of which showed a statistically significant difference compared with the blank control group [(100.9 +/- 16.8)%] (P > 0.05). However, after schistosomula soaked with siRNA-1 and siRNA-2 for 5 days, the levels of SjAR transcripts were reduced to (48.6 +/- 8.2)% and (73.4 +/- 4.7)%, respectively, both showing a statistically significant difference in comparison to the blank control group [(100.04 +/- 3.25)%] (P < 0.01). The siRNA-NC treated group (negative control) exhibited no statistically significant difference compared with the blank control group (P > 0.05). Accordingly, as shown by Western blotting, the SjAR protein levels of worms soaked with siRNA-1, siRNA-2 or siRNA-NC were 79.0%, 97.8% and 103.2%, respectively, compared with the blank control. CONCLUSION: RNAi treatment based on siRNA soaking reduce the expression of SjAR gene at both mRNA and protein level.

Chromosome-level genome assembly of the Asian citrus psyllid, Diaphorina citri.

Diaphorina citri is a global citrus pest. As a vector insect, it can transmit the causative agents of citrus huanglongbing, causing irreversible losses to the citrus industry. The acquisition of genomic information can provide a molecular genetic basis for effective control of D. citri. Here, the DNBSEQ™ , Oxford Nanopore Technologies, and Hi-C technologies are applied to generate a high-quality chromosome-level genome of D. citri. The genome size of D. citri was 523.78 Mb with a scaffold N50 of 47.05 Mb distributed on 13 chromosomes. A total of 250.64 Mb (47.85%) repeat sequences and 24 048 protein-coding genes were predicted. Genome resequencing of female and male individuals indicated that the sex chromosome system of D. citri is XO. Phylogenetic analysis demonstrated that D. citri and Pachypsylla venusta, which separated from their most recent common ancestor about 336.62 million years ago, were the most closely related. Additionally, we identified genes potentially involved in detoxification metabolism, pathogen transmission, and honeydew secretion for further investigation. The high-quality genome provides an important reference for developing effective management strategies of D. citri.

Identification and Functional Characterization of an Omega-Class Glutathione S-Transferase Gene PcGSTO1 Associated with Cyetpyrafen Resistance in Panonychus citri (McGregor).

Cyetpyrafen is a recently developed acaricide. The citrus red mite, Panonychus citri (McGregor), has developed significant resistance to cyetpyrafen. However, the molecular mechanism underlying the cyetpyrafen resistance in P. citri remains unclear. Glutathione S-transferases (GSTs) play a critical role in arthropod pesticide resistance. This study showed that GSTs were potentially related to the resistance of P. citri to cyetpyrafen through synergistic experiments and enzyme activity analysis. An omega-family GST gene, PcGSTO1, was significantly up-regulated in the egg, nymph, and adult stages of the cyetpyrafen-resistant strain. Additionally, silencing of PcGSTO1 significantly increased the mortality of P. citri to cyetpyrafen and recombinant PcGSTO1 demonstrated the ability to metabolize cyetpyrafen. Our results indicated that the overexpression of PcGSTO1 is associated with cyetpyrafen resistance in P. citri, and they also provided valuable information for managing resistance in P. citri.

Inertia-dependent dynamics of three-dimensional vesicles and red blood cells in shear flow.

A three-dimensional (3D) simulation study of the effect of inertia on the dynamics of vesicles and red blood cells (RBCs) has not been reported. Here, we developed a 3D model based on the front tracking method to investigate how inertia affects the dynamics of spherical/non-spherical vesicles and biconcave-shaped RBCs with the Reynolds number ranging from 0.1 to 10. The results showed that inertia induced non-spherical vesicles transitioned from tumbling to swinging, which was not observed in previous 2D models. The critical viscosity ratio of inner/outer fluids for the tumbling­swinging transition remarkably increased with an increasing Reynolds number. The deformation of vesicles was greatly enhanced by inertia, and the frequency of tumbling and tank-treading was significantly decreased by inertia. We also found that RBCs can transit from tumbling to steady tank-treading through the swinging regime when the Reynolds number increased from 0.1 to 10. These results indicate that inertia needs to be considered at moderate Reynolds number (Re ~ 1) in the study of blood flow in the human body and the flow of deformable particle suspension in inertial microfluidic devices. The developed 3D model provided new insights into the dynamics of RBCs under shear flow, thus holding great potential to better understand blood flow behaviors under normal/disease conditions.

Cytotoxic clerodane diterpenoids from the leaves and twigs of Casearia balansae.

Ten new clerodane diterpenoids (1-10), caseabalansins A-G, 18-epicaseabalansin A, 2-epicaseabalansin B, and 2-epicaseabalansin C, one new triterpenoid, balansinone (11), and seven known diterpenoids (12-18) were obtained from the leaves and twigs of Casearia balansae. Compounds 1 and 2 are the first examples of clerodane diterpenoids with an oxygen bridge between C-2 and C-19, and compounds 5-7 are three new diterpenoid artifacts presumably formed during the extraction process. The structures of the new compounds were established on the basis of extensive spectroscopic data, and that of 11 was verified by single-crystal X-ray crystallographic analysis. Compound 15 showed cytotoxic activity against the tumor cell lines PC3, DU145, SKOV3, and A549 with IC50 values of 4.5, 4.3, 5.1, and 5.7 µM, respectively. Compounds 1a, 2a, and 4 showed selective activity against PC3 tumor cells.

[Corynebacterium aquaticum infection: one case report and literature review].

OBJECTIVE: To study the diagnosis and treatment of Corynebacterium aquaticum infection. METHODS: A retrospective analysis of one case of Corynebacterium aquaticum infection and literature review were conducted. RESULTS: A 39-year old male patient was admitted because of cough, sputum production, fever and right chest pain for 10 days. Broad-spectrum antibiotic therapy had been given in another hospital, but the patient's condition had deteriorated.Nuclear magnetic resonance-guided percutancous transthoracic needle aspiration and lung tissue, pleural fluid and blood culture were performed after admission to our hospital. Corynebacterium aquaticum was grown from the lung tissue, the blood and the pleural effusion. Therefore the diagnosis of Corynebacterium aquaticum pneumonia complicated with pyothorax and septicemia was confirmed. After draining of pleural pus and intravenous vancomycin therapy, the patient recovered and was discharged from hospital. After literature search, we did not find reports on Corynebacterium aquaticum pneumonia complicated with pyothorax and septicemia. CONCLUSIONS: Corynebacterium aquaticum pneumonia complicated with pyothorax and septicemia is rare. The diagnosis could be confirmed by bacterial culture of lung tissue, pleural fluid and blood. Thoracic cavity draining and intravenous vancomycin are effective therapies for the disease.

Enhanced Mitophagy in Cholesteatoma Epithelial Cells.

HYPOTHESIS: Mitophagy may have a potential role in the pathogenesis of acquired cholesteatoma. BACKGROUND: Enhanced mitophagy has been proven to be involved in various cancers. However, its role in the pathogenesis of cholesteatoma, which shares some common features with cancer, is controversial. This study investigated mitophagy in cholesteatoma epithelial cells. METHODS: The autophagy protein markers LC3-II and p62 and mitophagy proteins BNIP3, Parkin, and PINK1 were analyzed in cholesteatoma epithelial cells and external auditory canal epithelium cells by immunoblotting. The results were confirmed by immunohistochemistry. Adenovirus Ad-mCherry-GFP-LC3B and Ad-GFP-LC3B were used to evaluate autophagic activity. Transmission electron microscopy was used to observe and analyze autophagosomes. RESULTS: LC3-II expression was increased in cholesteatoma cells, whereas soluble and insoluble p62 levels were decreased. The expressions of BNIP3, Parkin, and PINK1 were higher in total protein and mitochondrial protein of cholesteatoma cells compared with normal external auditory canal epithelium cells. Autophagic activity was increased in cholesteatoma cells compared with normal external auditory canal epithelium cells. CONCLUSION: Mitophagy was enhanced in cholesteatoma epithelial cells and may have a potential role in the pathogenesis of acquired cholesteatoma.