A networked iron and nitrogen-doped ZIF-8/MWCNTs heterostructure for oxygen reduction reaction.

Zeolitic Imidazolate Frameworks-8 (ZIF-8) is commonly used as an ideal precursor for non-noble metal catalysts because of its high specific surface area, ultra-high porosity, and N-rich content. Upon pyrolyzing ZIF-8 at 900 °C in Ar, the resulting material, referred to as Z8, displayed good activity toward the oxygen reduction reaction (ORR). Then the ZIF-8 was mixed with various conductive carbon materials, such as multiwall carbon nanotubes (MWCNTs), Acetylene black (ACET), Vulcan XC-72R (XC-72R), and Ketjenblack EC-600JD (EC-600JD), to form Z8 composites. The Z8/MWCNTs composite exhibited enhanced ORR activity owing to its network structure, meso-/microporous hierarchical porous structure, improved electrical conductivity, and graphitization. Subsequently, iron and nitrogen co-doping is achieved through the pyrolysis of a mixture comprising Fe, N precursor, and ZIF-8/MWCNTs, which is denoted as FeN-Z8/MWCNTs. The intrinsically high electrical conductivity of MWCNTs facilitated efficient electron transfer during the ORR, while the meso-/microporous hierarchical porous structure and network structure of Fe, N co-doped ZIF-8/MWCNTs promoted oxygen transport. The presence of Fe-containing species in the catalyst acted as activity centers for ORR. This strategy of preparing Z8 composites and modifying them with Fe, N co-doping offers an insightful approach to designing cost-effective electrocatalysts.

A new target of radiotherapy combined with immunotherapy: regulatory T cells.

Radiotherapy is one important treatment for malignant tumours. It is widely believed today that radiotherapy has not only been used as a local tumour treatment method, but also can induce systemic anti-tumour responses by influencing the tumour microenvironment, but its efficacy is limited by the tumour immunosuppression microenvironment. With the advancement of technology, immunotherapy has entered a golden age of rapid development, gradually occupying a place in clinical tumour treatment. Regulatory T cells (Tregs) widely distributing in the tumour microenvironment play an important role in mediating tumour development. This article analyzes immunotherapy, the interaction between Tregs, tumours and radiotherapy. It briefly introduces immunotherapies targeting Tregs, aiming to provide new strategies for radiotherapy combined with Immunotherapy.

The efficacy of auto-cross-linked hyaluronic acid gel in addition to oestradiol and intrauterine balloon insertion in the prevention of adhesion reformation after hysteroscopic adhesiolysis.

RESEARCH QUESTION: What is the efficacy of auto-cross-linked hyaluronic acid gel use in preventing adhesion reformation after intrauterine adhesiolysis? DESIGN: This was a single-centre, double-blind randomized controlled trial. RESULTS: In total 171 participants successfully completed the study (84 in the treatment group and 87 in the control group). There was no significant difference in pre-operative variables between the two groups. The primary outcome measure was the adhesion reformation rate at second-look and third-look hysteroscopy. At second-look hysteroscopy, there was no significant difference in adhesion recurrence rate between the treatment group (20.2%, 17/84) and the control group (23.0%, 20/87; P = 0.662). At third-look hysteroscopy, there was also no significant difference in adhesion recurrence rate between the treatment group (9.5%, 8/84) and the control group (11.5%, 10/87; P = 0.675). The secondary outcome measure was the median American Fertility Society (AFS) score, which was not significantly different at second-look hysteroscopy 4 weeks after surgery between the treatment group (0, range 0-4.0) and the control group (0, range 0-4.0; P = 0.475), and at third-look hysteroscopy 8 weeks after surgery between the treatment group (0, range 0-3.5) and the control group (0, range 0-4.0; P = 0.965). Regarding the menstrual flow improvement rate 3 months post-operatively, there was no significant difference between the treatment and control groups (67.9% versus 64.4%; P = 0.630). CONCLUSIONS: The application of auto-cross-linked hyaluronic acid gel does not seem to reduce the incidence and severity of intrauterine adhesion recurrence or affect the menstrual pattern after hysteroscopic removal of mild to moderate intrauterine adhesions.

The Effect of Estrogen in the Prevention of Adhesion Reformation after Hysteroscopic Adhesiolysis: A Prospective Randomized Control Trial.

STUDY OBJECTIVE: To evaluate whether estrogen therapy can reduce adhesion reformation after hysteroscopic adhesiolysis. DESIGN: A single-center, single blinded, randomized controlled trial. SETTING: A tertiary University Hospital. PATIENTS: A total of 207 patients with mild (American Fertility Society [AFS] score 1-6) and severe (AFS score 7-12) intrauterine adhesion who underwent hysteroscopic adhesiolysis. INTERVENTIONS: Patients were randomized to a treatment group or a control group, stratified according to the preoperative AFS adhesion score. The treatment group received estrogen, and the control group did not. All patients had second-look hysteroscopy at 4 weeks and third-look hysteroscopy at 8 weeks after surgery. MEASUREMENTS AND MAIN RESULTS: Primary outcome measures were adhesion reformation rate and AFS score at third-look hysteroscopy. Secondary outcome measures included adhesion reformation rate and AFS score at second-look hysteroscopy and menstrual pattern improvement rate at 3 months after operation. Among subjects with mild intrauterine adhesion, there was no significant difference between the treatment group and control group with regard to adhesion reformation rate at third-look hysteroscopy (10.6% vs 13.6%), AFS score (mean ± standard deviation) at third-look hysteroscopy (1.1 ± 1.2 vs 1.3 ± 1.2), and menstrual pattern improvement rate at 3-month follow-up (89.4% vs 86.4%). Similarly, among those with severe intrauterine adhesion, there was no significant difference between the treatment group and control group in adhesion reformation rate at third-look hysteroscopy (32.6% vs 26.7%), AFS score (mean ± standard deviation) at third-look hysteroscopy (2.5±2.2 vs 2.7±2.1), and menstrual pattern improvement rate at 3-month follow-up (84.8% vs 73.3%). CONCLUSION: Postoperative estrogen therapy did not appear to reduce the incidence or severity of adhesion reformation, nor did it improve the menstrual pattern, regardless of whether the pre-existing intrauterine adhesion was mild or severe.

Discovery of STAT3 and Histone Deacetylase (HDAC) Dual-Pathway Inhibitors for the Treatment of Solid Cancer.

Nowadays, simultaneous inhibition of multiple targets through drug combination is an important anticancer strategy owing to the complex mechanism behind tumorigenesis. Recent studies have demonstrated that the inhibition of histone deacetylases (HDACs) will lead to compensated activation of a notorious cancer-related drug target, signal transducer and activator of transcription 3 (STAT3), in breast cancer through a cascade, which probably limits the anti-proliferation effect of HDAC inhibitors in solid tumors. By incorporating the pharmacophore of the HDAC inhibitor SAHA (vorinostat) into the STAT3 inhibitor pterostilbene, a series of potent pterostilbene hydroxamic acid derivatives with dual-target inhibition activity were synthesized. An excellent hydroxamate derivate, compound 14, inhibited STAT3 (KD = 33 nM) and HDAC (IC50 = 23.15 nM) with robust potency in vitro. Compound 14 also showed potent anti-proliferation ability in vivo and in vitro. Our study provides the first STAT3 and HDAC dual-target inhibitor for further exploration.

The association of chronic endometritis with mid-trimester loss due to cervical incompetence and the outcome of laparoscopic cervical cerclage.

PURPOSE: To examine the association of chronic endometritis (CE) with cervical incompetence (CI) in Chinese women with mid-trimester loss, and the impact of the presence of CE on the outcome of laparoscopic cervical cerclage (LCC). METHODS: This retrospective cohort study included a study group of 293 women with mid-trimester loss due to CI (group I) and a comparison group of 332 women with recurrent first-trimester miscarriage (group II). Immunohistochemical study using CD138 epitope for the diagnosis of CE was completed in all subjects. Pre-conception LCC was undertaken in 247 women in the study group (group I). The study was approved by Institutional Review Board (IRB) (number 2015FXHEC-KY005). RESULTS: The prevalence of CE in group I was 42%, significantly (P < 0.001) higher than that of 23.5% in group II. Among 247 women in group I, there were no significant difference in mid-trimester loss rate, preterm delivery rate and term delivery rate in women with and without CE (2.2, 12.0, 85.8% vs. 1.8, 10.1, 88.1% respectively) and between women with CE treated and not treated with antibiotics prior to conception (2.3, 9.3, 88.4% vs. 2.0, 14.3, 83.7% respectively). CONCLUSIONS: Mid-trimester loss due to cervical incompetence is associated with chronic endometritis; However, the presence or not of CE and whether it was treated with antibiotics prior to conception did not appear to significantly influence the obstetric outcomes of women with CI after LCC.

Impact of antibiotic therapy on the rate of negative test results for chronic endometritis: a prospective randomized control trial.

OBJECTIVE: To compare the rates of negative test results for chronic endometritis (CE) between subjects who did and did not receive antibiotic treatment. DESIGN: Prospective, single-blind randomized controlled trial. SETTING: Tertiary hysteroscopic center in a university teaching hospital. PATIENT(S): A total of 132 women with CE confirmed with immunohistochemical study with CD138 epitope. INTERVENTION(S): Women randomized to antibiotic therapy received oral levofloxacin 500 mg and tinidazole 1,000 mg daily for 14 days. Women randomized to the control group did not receive any treatment. A repeated endometrial biopsy was performed 4 to 8 weeks after the initial biopsy to determine whether CE was still present. MAIN OUTCOME MEASURE(S): The rate of negative test results for CE (from positive to negative). RESULT(S): The CE rate of negative test results in the treatment group (89.3%) after one course of antibiotic treatment was significantly higher than that in the control group (12.7%). Among subjects who attempted pregnancy, there was no significant difference in ongoing pregnancy rates and miscarriage rates between the treatment arm (43.2%, 5.4%) and the control arm (25.7%, 14.3%). Among subjects randomized, there was also no significant difference in ongoing pregnancy rates and miscarriage rates between the treatment arm (27.1%, 3.4%) and the control arm (16.4%, 9.1%). CONCLUSION: A course of broad-spectrum oral antibiotic therapy for 14 days is effective in the treatment of CE in >89.8% of cases. However, it is not yet clear whether treatment improved pregnancy outcomes. CLINICAL TRIAL IDENTIFICATION NUMBER: NCT02648698.

CircPlekha7 plays an anti-fibrotic role in intrauterine adhesions by modulating endometrial stromal cell proliferation and apoptosis.

Circular RNA (circRNA) plays a key role in the development and progression of several diseases; however, its role in intrauterine adhesions (IUAs) is not well understood. This study aims to investigate the expression profiles and potential role of circRNA in IUA. RNA-sequencing was performed to screen for abnormally expressed circRNAs in TGF-ß1-induced IUA endometrial stromal cell (ESC) model (IUA group) and an SMAD3 inhibitor, SIS3-treated IUA ESC model (SIS3 group). Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses were performed to uncover the key functions and pathways. Interaction networks were constructed and analyzed based on the competing endogenous RNA hypothesis of circRNA. CircRNAs were validated by Sanger sequencing and quantitative polymerase chain reaction (qPCR). Cell proliferation and apoptosis were measured using MTS and flow cytometry, respectively. The protein and mRNA expression levels of fibrosis-related proteins were measured using western blotting and reverse transcription-qPCR, respectively. A total of 66 circRNAs were differentially expressed between the IUA and SIS3 groups. CircPlekha7 was identified as one of the significantly upregulated circRNAs in the SIS3 group. Overexpression of circPlekha7 enhanced apoptosis, decreased the viability of ESCs, and suppressed the expression of α-SMA, collagen I, and SMAD3 in ESCs; whereas knockdown of circPlekha7 exhibited opposite results. Altogether, the results indicate that circPlekha7 plays an anti-fibrotic role in IUA and may serve as a promising prognostic biomarker for patients with IUA. Therefore, overexpression of circPlekha7 could be a potential treatment strategy for IUA.

Functional analysis of miRNAs combined with TGF-ß1/Smad3 inhibitor in an intrauterine rat adhesion cell model.

In this study, we aimed to study the role of miRNAs in intrauterine adhesion (IUA) disease. An IUA cell model was constructed by TGF-ß1. Smad3 inhibitor (SIS3) can inhibit the Smad3 signaling pathway and affect the role of TGF-ß1; thus, it was used to identify the role of Smad3 and related miRNAs in IUA. Cell number significantly increased in the TGF-ß1 group after 72 h and 96 h, respectively, compared with that in the control group (P < 0.05). However, cell proliferation was significantly decreased in the TGF-ß1 + SIS3 group (P < 0.0001). Cell apoptosis was increased in the TGF-ß1 + SIS3 group compared with that in the TGF-ß1 group. Western Blot (WB) analysis suggested that TGF-ß1 treatment could effectively increase the expression of α-SMA, COL1, Smad3, and p-Smad3, which could be inhibited by SIS3 treatment. A total of 235 and 530 differentially expressed miRNAs in the TGF-ß1 + SIS3 group were significantly up- and downregulated compared with those in the TGF-ß1 group, respectively. These differentially expressed miRNAs were enriched in the MAPK and PI3K-AKT pathways. The ten most differentially expressed miRNAs were selected to verify their expressions using quantitative real-time polymerase chain reaction (qPCR). Furthermore, overexpression of rno-miR-3586-3p and rno-miR-455-5p can promote cell proliferation and exacerbate the IUA pathogenic process. However, overexpression of rno-miR-204-3p and rno-miR-3578 can inhibit cell behavior and IUA progression. The above results can provide detailed information for the understanding of IUA molecular mechanisms.

Discovery of bazedoxifene analogues targeting glycoprotein 130.

Deregulation of GP130 in signal transduction is involved in multiple types of human diseases, especially in cancers, indicating that GP130 is an attractive target for cancer therapy. However, GP130 was conventionally considered as an undruggable target thus the discovery of GP130 PPI inhibitors is extremely challenging. By the aid of structure-based drug design, in this study, two series of bazedoxifene based analogues were designed to target GP130 D1 domain and block the IL-6/GP130/STAT3 signaling pathway for antitumor treatment. Most of these designed compounds displayed potent anti-proliferative activity against cancer cells. The representative compound 10a was demonstrated to directly bind to GP130 protein with an affinity (KD) value of 3.8 µM via both SPR and DARTS methods. Subsequently, molecular docking study predicted that 10a targeted D1 domain of GP130 and co-IP assay demonstrated that 10a did not inhibit IL-6R/GP130 interaction, which meant 10a did not bind to the D2 and D3 domains of GP130. Moreover, 10a selectively inhibited JAK2 and STAT3 phosphorylation as well as IL-6 induced STAT3 phosphorylation. 10a effectively inhibited tumor cell viability, migration and promoted apoptosis. Furthermore, 10a effectively suppressed xenograft model tumor growth in vivo. Taken together, this study described a new class of bazedoxifene derived GP130 inhibitors as antitumor agents.

Durations of intrauterine balloon therapy and adhesion reformation after hysteroscopic adhesiolysis: a randomized controlled trial.

RESEARCH QUESTION: Does placing an intrauterine balloon for different durations (7, 14 or 28 days) affect the recurrence of intrauterine adhesions after hysteroscopic adhesiolysis? DESIGN: Prospective randomized control trial involving 138 patients recruited over a 12-month period and followed up post-operatively for 15 months. The primary outcome measure was the rate of adhesion reformation at third-look hysteroscopy. RESULTS: At third-look hysteroscopy, 8 weeks after the initial hysteroscopy, the adhesion recurrence rate in women who had an intrauterine balloon for 28 days (20%) was significantly (P < 0.01) lower than that of women who had the balloon for 14 days (55%) or 7 days (36.8%). CONCLUSIONS: Placing an intrauterine balloon for 28 days instead of 7 or 14 days after hysteroscopic adhesiolysis resulted in a greater reduction in the recurrence rate of adhesions. However, the study was underpowered to address whether the ongoing pregnancy rate could be improved by keeping the balloon in the uterine cavity for a longer period of time.

Real-time monitoring of etoposide prodrug activated by hydrogen peroxide with improved safety.

Etoposide is one of the most used first-line chemotherapeutic drugs. However, its application is still limited by its side effects. Herein, we designed a novel H2O2 sensitive prodrug 6YT for selectively releasing the anti-cancer drug etoposide in cancer cells. In this paper, etoposide and a hydrogen peroxide (H2O2) sensitive aryl borate ester group were linked by a fluorescent coumarin and finally the prodrug 6YT was generated. The fluorescence of coumarin was quenched before the connected aryl borate ester group was cleaved by H2O2. However, in the high level H2O2 environment of the tumor, the fluorescence could be activated simultaneously with the release of etoposide, and the drug release state of the prodrug was monitored real-time. With the support of 6YT, we obtained direct and visual evidence of etoposide release in a high H2O2 environment both in cells and zebrafish. The prodrug 6YT was also verified with comparable activity and improved safety with etoposide both in cells and in a mouse model. As a safe and effective prodrug, 6YT is expected to be one of the promising candidates in chemotherapy against cancer.

Discovery of fluorescent coumarin-benzo[b]thiophene 1, 1-dioxide conjugates as mitochondria-targeting antitumor STAT3 inhibitors.

STAT3 has been extensively studied as a potential antitumor target. Though studies on regulating STAT3 mainly focus on the inhibition of STAT3 phosphorylation at Tyr705 residue, the phosphorylation at Ser727 residue of STAT3 protein is also closely associated with the mitochondrial import of STAT3 protein. N, N-diethyl-7-aminocoumarin is a fluorescent mitochondria-targeting probe. In this study, a series of STAT3 inhibitors were developed by connecting N, N-diethyl-7-aminocoumarin fluorophore with benzo [b]thiophene 1, 1-dioxide moiety. All designed compounds displayed potent anti-proliferative activity against cancer cells. The representative compound 7a was mainly accumulated in mitochondria visualized by its fluorescence. STAT3 phosphorylation was inhibited by compound 7a at both Tyr705 and Ser727 residues. Compound 7a inhibited STAT3 phosphorylation whereas had no influence on the phosphorylation levels of STAT1, JAK2, Src and Erk1/2, indicating good selectivity of compound 7a. Moreover, compound 7a down-regulated the expression of STAT3 target genes Bcl-2 and Cyclin D1, increased ROS production and remarkably reduced the mitochondrial membrane potential to induce mitochondrial apoptotic pathway. Furthermore, compound 7ain vivo suppressed breast cancer 4T1 implanted tumor growth. Taken together, these results highlighted that compound 7a might be a promising mitochondria-targeting STAT3 inhibitor for cancer therapy.

Correlation between hysteroscopy findings and chronic endometritis.

OBJECTIVE: To evaluate the role of hysteroscopy in the diagnosis of chronic endometritis (CE). DESIGN: Retrospective cohort study. SETTING: University teaching hospital. PATIENT(S): A consecutive series of 1,189 cases of diagnostic hysteroscopy. INTERVENTION(S): Endometrial biopsy specimens were obtained after hysteroscopy for routine histology and immunohistochemistry for plasma cells using a CD138 epitope. MAIN OUTCOME MEASURE(S): Observer variability, sensitivity, specificity, and accuracy of the hysteroscopic features in the diagnosis of CE. RESULT(S): Immunohistochemistry of biopsy specimens showed presence of CD138 cells in 322 of 1,189 cases (27.1%). Among cases that tested positive for CD138 cells, the prevalence of hysteroscopic features was as follows: endometrial hyperemia, 169 of 322 (52.5%); endometrial interstitial edema, 27 of 322 (8.4%); and micro-polyps, 11 of 322 (3.4%). The κ value of intraobserver and interobserver agreement on the presence or absence of the hysteroscopic feature of CE was 0.86 and 0.73, respectively. The sensitivity, specificity, positive and negative predictive value, and diagnostic accuracy of the presence of one or more hysteroscopy features were 59.3%, 69.7%, 42.1%, 82.8%, and 66.9%, respectively. CONCLUSION(S): The finding of endometrial hyperemia, micro-polyps or endometrial interstitial edema during hysteroscopy should alert to the diagnosis of CE, but the overall accuracy of hysteroscopic examination with regard to the diagnosis of CE is only 67%, so it should not be used to replace histologic examination as the diagnostic tool of choice. In women in whom a diagnosis of CE is considered likely, endometrial biopsy should be obtained to examine plasma cells by immunohistochemistry, which should remain the preferred method for diagnosis.

Discovery of new benzensulfonamide derivatives as tripedal STAT3 inhibitors.

Persistent activated STAT3 has a striking correlation with cancer development and inhibition of STAT3 signaling pathway is a novel therapeutic way for human cancers. Among STAT family, STAT1 and STAT3 play opposite roles in tumorigenesis. However, the discovery of selective STAT3 inhibitors is still challenging to date. In this study, a series of small-molecular (MW < 500) benzensulfanilamide derivatives were designed to selectively suppress STAT3 activation for anti-cancer treatment. The most potent compound 11 inhibited both overexpressed and IL-6 induced STAT3 phosphorylation, whereas 11 displayed little effect on the phosphorylation of other STAT isoforms STAT1, STAT5, demonstrating 11 was a selective STAT3 inhibitor. Meanwhile, 11 dismissed STAT3 DNA binding activity and colony formation. In addition, 11 elevated the ROS level and induced apoptosis of cancer cells. Furthermore, 11 effectively suppressed tumor growth in an in vivo mouse-xenograft model.

Prevalence and confounders of chronic endometritis in premenopausal women with abnormal bleeding or reproductive failure.

In this retrospective cohort study, a consecutive series of 1551 premenopausal women underwent hysteroscopy and endometrial biopsy. Chronic endometritis was diagnosed when plasma cell in endometrial tissue was detected by immunohistochemistry using CD138 epitope. The overall prevalence of chronic endometritis in the population studied was 24.4% The prevalence was significantly increased in the following conditions: recurrent implantation failure (40.8%; P < 0.001), abnormal uterine bleeding (40.7 %; P < 0.001), endometrial hyperplasia (50.0%, P < 0.05) and submucosal fibroid (59.1%; P < 0.001) than those without the respective conditions. The prevalence in specimens obtained from the proliferative phase (26.0%) was significantly higher (P < 0.05) than those from the luteal phase (17.5%). Logistic regression analysis showed three significant factors affecting the prevalence, in descending order of importance: clinical presentation, endometrial hyperplasia and stage of the cycle from which the specimen was obtained. The confounding variables identified in this study may account for the wide range of published prevalence of the condition, and should be considered in the analysis of prevalence data relating to chronic endometritis.

SIRT1 exhibits antioxidative effects in HT22 cells induced by tert-butyl alcohol.

Tertiary butyl alcohol (TBA) is a principal metabolite of methyl tertiary-butyl ether (MTBE), a common pollutant worldwide in the ground or underground water, which is found to produce nervous system damage. Nevertheless, few data regarding the effects of TBA has been reported. Studies indicated that oxidative stress plays a pivotal role in MTBE neurotoxic mechanism. Sirtuin 1 (SIRT1) has been reported to exert a neuroprotective effect on various neurologic diseases via resistance to oxidative stress by deacetylating its substrates. In this study, we examined levels of oxidative stress after exposure to TBA for 6 h in HT22 cells and HT22 cells with SIRT1 silencing (transfected with SIRT1 siRNA) or high expression (preconditioned with agonists SRT1720). We found that TBA activated oxidative stress by increasing generation of intracellular reactive oxygen species (ROS), malondialdehyde (MDA) and Oxidized glutathione (GSSG), and decreasing contents of superoxide dismutase (SOD) and glutathione reductase (GSH). In additional, levels of TBA-induced oxidative stress were aggravated when SIRT1 silenced but alleviated when SIRT1 enhanced. Our study indicated that SIRT1 mitigated oxidative stress induced by TBA.

FOXO3 promoted mitophagy via nuclear retention induced by manganese chloride in SH-SY5Y cells.

OBJECTIVES: To evaluate the role of FOXO3 during the process of mitophagy induced by manganese chloride (MnCl2), mitochondrial dysfunction and mitophagy were detected before and after FOXO3 was knocked down in SH-SY5Y cells. METHOD: Transmission electron microscopy (TEM), flow cytometry, confocal microscopy and a western blot were used to detect mitochondrial ultrastructure and autophagy, Ca2+ levels, mitochondrial reactive oxygen species (ROS) and the mitochondrial membrane potential (MMP), autophagosomes and mitophagy marker proteins (p62, LC3-II/LC3-I, Beclin-1, PINK1 and P-parkin), respectively. RESULTS: After SH-SY5Y cells were exposed to MnCl2, the levels of cytoplasmic Ca2+ and mitochondrial ROS increased but the mitochondrial MMP decreased significantly compared to the control in a dose- and time-dependent manner (p < 0.05), which indicated that MnCl2 can lead to mitochondrial dysfunction. Under TEM, mitophagy and autolysosomes were observed. The WB results also showed that mitophagy marker proteins including LC3-II/LC3-I, Beclin-1, PINK1 and P-parkin except for p62 increased in a dose- and time-dependent manner, accompanied by FOXO3 nuclear retention, which indicated that MnCl2 can lead to mitophagy and FOXO3 nuclear translocation may be involved in this process. After FOXO3 was knocked down, the inverse results of mitophagy and the levels of mitochondrial ROS decreasing were observed, which showed that FOXO3 silencing could inhibit mitophagy and mitochondrial dysfunction induced by MnCl2. CONCLUSIONS: Our results indicated that Mn could induce mitophagy by enhancing FOXO3 nuclear retention, which might promote mitophagy induced by MnCl2.