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Mammalian orthoreovirus (MRV) outer capsid protein σ3 is a multifunctional protein containing a double-stranded RNA-binding domain, which facilitates viral entry and assembly. We reasoned that σ3 has an innate immune evasion function. Here, we show that σ3 protein localizes in the mitochondria and interacts with mitochondrial antiviral signaling protein (MAVS) to activate the intrinsic mitochondria-mediated apoptotic pathway. Consequently, σ3 protein promotes the degradation of MAVS through the intrinsic caspase-9/caspase-3 apoptotic pathway. Moreover, σ3 protein can also inhibit the expression of the components of the RNA-sensing retinoic acid-inducible gene (RIG)-like receptor (RLR) signaling pathway to block antiviral type I interferon responses. Mechanistically, σ3 inhibits RIG-I and melanoma differentiation-associated gene 5 expression is independent of its inhibitory effect on MAVS. Overall, we demonstrate that the MRV σ3 protein plays a vital role in negatively regulating the RLR signaling pathway to inhibit antiviral responses. This enables MRV to evade host defenses to facilitate its own replication providing a target for the development of effective antiviral drugs against MRV. IMPORTANCE: Mammalian orthoreovirus (MRV) is an important zoonotic pathogen, but the regulatory role of its viral proteins in retinoic acid-inducible gene-like receptor (RLR)-mediated antiviral responses is still poorly understood. Herein, we show that MRV σ3 protein co-localizes with mitochondrial antiviral signaling protein (MAVS) in the mitochondria and promotes the mitochondria-mediated intrinsic apoptotic pathway to cleave and consequently degrade MAVS. Furthermore, tryptophan at position 133 of σ3 protein plays a key role in the degradation of MAVS. Importantly, we show that MRV outer capsid protein σ3 is a key factor in antagonizing RLR-mediated antiviral responses, providing evidence to better unravel the infection and transmission mechanisms of MRV.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal , Proteínas do Capsídeo , Orthoreovirus de Mamíferos , Transdução de Sinais , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas do Capsídeo/metabolismo , Proteínas do Capsídeo/genética , Humanos , Orthoreovirus de Mamíferos/genética , Animais , Apoptose , Proteína DEAD-box 58/metabolismo , Proteína DEAD-box 58/genética , Mitocôndrias/metabolismo , Imunidade Inata , Camundongos , Evasão da Resposta Imune , Células HEK293 , Receptores Imunológicos/metabolismo , Helicase IFIH1 Induzida por Interferon/metabolismo , Helicase IFIH1 Induzida por Interferon/genética , Linhagem Celular , Interações Hospedeiro-PatógenoRESUMO
Mammalian reovirus (MRV) is a non-enveloped, gene segmented double-stranded RNA (dsRNA) virus. It is an important zoonotic pathogen that infects many mammals and vertebrates that act as natural hosts and causes respiratory and digestive tract diseases. Studies have reported that RIG-I and MDA5 in the innate immune cytoplasmic RNA-sensing RIG-like receptor (RLR) signaling pathway can recognize dsRNA from MRV and promote antiviral type I interferon (IFN) responses. However, the mechanism by which many MRV-encoded proteins evade the host innate immune response remains unclear. Here, we show that exogenous µ1 protein promoted the proliferation of MRV in vitro, while knockdown of MRV µ1 protein expression by shRNA could impair MRV proliferation. Specifically, µ1 protein inhibited MRV or poly(I:C)-induced IFN-ß expression, and attenuated RIG-I/MDA5-mediated signaling axis transduction during MRV infection. Importantly, we found that µ1 protein significantly decreased IFN-ß mRNA expression induced by MDA5, RIG-I, MAVS, TBK1, IRF3(5D), and degraded the protein expression of exogenous MDA5, RIG-I, MAVS, TBK1 and IRF3 via the proteasomal and lysosomal pathways. Additionally, we show that µ1 protein can physically interact with MDA5, RIG-I, MAVS, TBK1, and IRF3 and attenuate the RIG-I/MDA5-mediated signaling cascades by blocking the phosphorylation and nuclear translocation of IRF3. In conclusion, our findings reveal that MRV outer capsid protein µ1 is a key factor in antagonizing RLRs signaling cascades and provide new strategies for effective prevention and treatment of MRV infection.
Assuntos
Proteínas do Capsídeo , Proteína DEAD-box 58 , Fator Regulador 3 de Interferon , Helicase IFIH1 Induzida por Interferon , Orthoreovirus de Mamíferos , Receptores Imunológicos , Transdução de Sinais , Animais , Humanos , Transporte Ativo do Núcleo Celular , Núcleo Celular/metabolismo , Proteína DEAD-box 58/metabolismo , Células HEK293 , Imunidade Inata/imunologia , Fator Regulador 3 de Interferon/metabolismo , Interferon beta/metabolismo , Interferon beta/imunologia , Helicase IFIH1 Induzida por Interferon/metabolismo , Helicase IFIH1 Induzida por Interferon/genética , Orthoreovirus de Mamíferos/imunologia , Orthoreovirus de Mamíferos/fisiologia , Fosforilação , Proteínas Serina-Treonina Quinases , Infecções por Reoviridae/imunologia , Transdução de Sinais/imunologia , Proteínas Virais/metabolismo , Proteínas do Capsídeo/metabolismoRESUMO
OBJECTIVE: The purpose of this study was to explore the therapeutic characteristics of mesenchymal stem cells generated from human umbilical cord (hUC-MSCs) when utilized in conjunction with auto-crosslinked hyaluronic acid gel (HA-gel) for the management of intrauterine adhesion (IUA). The goal was to see how this novel therapy could enhance healing and improve outcomes for IUA patients. METHODS: In this study, models of intrauterine adhesion (IUA) were established in Sprague-Dawley (SD) rats, which were then organized and divided into hUC-MSCs groups. The groups involved: hUC-MSCs/HA-gel group, control group, and HA-gel group. Following treatment, the researchers examined the uterine cavities and performed detailed analyses of the endometrial tissues to determine the effectiveness of the interventions. RESULTS: The results indicated that in comparison with to the control group, both HA-gel, hUC-MSCs, and hUC-MSCs/HA-gel groups showed partial repair of IUA. However, in a more notable fashion transplantation of hUC-MSCs/HA-gel complex demonstrated significant dual repair effects. Significant outcomes were observed in the group treated with hUC-MSCs and HA-gel, they showed thicker endometrial layers, less fibrotic tissue, and a higher number of endometrial glands. This treatment strategy also resulted in a significant improvement in fertility restoration, indicating a profound therapeutic effect. CONCLUSIONS: The findings of this study suggest that both HA-gel, hUC-MSCs, and hUC-MSCs/HA-gel complexes have the potential for partial repair of IUA and fertility restoration caused by endometrium mechanical injury. Nonetheless, the transplantation of the hUC-MSCs/HA-gel complex displayed exceptional dual healing effects, combining effective anti-adhesive properties with endometrial regeneration stimuli.
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Ácido Hialurônico , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , Ratos Sprague-Dawley , Cordão Umbilical , Doenças Uterinas , Ácido Hialurônico/farmacologia , Ácido Hialurônico/química , Feminino , Animais , Transplante de Células-Tronco Mesenquimais/métodos , Humanos , Ratos , Aderências Teciduais , Cordão Umbilical/citologia , Doenças Uterinas/terapia , Géis , Endométrio/efeitos dos fármacos , Endométrio/citologia , Modelos Animais de DoençasRESUMO
Histone H1.2 is a member of the linker histone family, which plays extensive and crucial roles not only in the regulation of chromatin dynamics, cell cycle, and cell apoptosis, but also in viral diseases and innate immunity response. Recently, it was discovered that H1.2 regulates interferon-ß and inhibits influenza virus replication, whereas its role in other viral infections is poorly reported. Here, we first found the up-regulation of H1.2 during Encephalomyocarditis virus (EMCV) infection, implying that H1.2 was involved in EMCV infection. Overexpression of H1.2 inhibited EMCV proliferation, whereas knockdown of H1.2 showed a significant promotion of virus infection in HEK293T cells. Moreover, we demonstrated that overexpression of H1.2 remarkably enhanced the production of EMCV-induced type I interferon, which may be the crucial factor for H1.2 proliferation-inhibitory effects. We further found that H1.2 up-regulated the expression of the proteins of the MDA5 signaling pathway and interacted with MDA5 and IRF3 in EMCV infection. Further, we demonstrated that H1.2 facilitated EMCV-induced phosphorylation and nuclear translocation of IRF3. Briefly, our research uncovers the mechanism of H1.2 negatively regulating EMCV replication and provides new insight into antiviral targets for EMCV.
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Vírus da Encefalomiocardite , Histonas , Humanos , Células HEK293 , Imunidade Inata , Helicase IFIH1 Induzida por Interferon/genética , Transdução de Sinais , Replicação ViralRESUMO
OBJECTIVE: Existing studies have conflicting results about the predictors of forgotten joints in patients with total knee arthroplasty (TKA), and the relationship between psychosocial factors and forgotten knees is unknown. The purpose of this study was to confirm predictors for the forgotten joint in TKA patients. METHODS: This was an observational, prospective longitudinal study. A total of 205 patients who underwent TKA and a 6-month follow-up were included between August 2020 and September 2021. Demographic characteristics, clinical characteristics, and psychosocial variables were collected before TKA surgery (T0). The forgotten joint score (FJS) was taken before TKA surgery (T0) and at 1 month (T1), 3 months (T3), and 6 months (T6) after TKA surgery. The psychosocial variables were also completed at T6. Bivariate and multivariable linear regressions (LR) were performed to screen the predictors associated with FJS (T6). RESULTS: Patients who underwent TKA in our study had a mean FJS of 20.3 ± 12.2 before surgery, 15.9 ± 10.3 at 1 month, 28.7 ± 12.6 at 3 months, and 40.3 ± 12.5 at 6 months. The predictors were sex, combined musculoskeletal disorders (MSD), operation time, FJS (T3), range of motion (ROM) (T6), pain score (T6), Groningen orthopaedic social support scale (GO-SSS) score (T6), and the generalized anxiety disorder scale (GAD) score (T6). The data satisfied the assumptions of multivariable linear regressions. The multiple R2 of LR was 0.71, and the adjusted R2 was 0.70. The F-statistic of the LR model was 59.5 (p < 0.000). CONCLUSION: Our study revealed the level of forgotten knee decreased slightly from preoperation to 1 month postoperatively and then increased from 1 month postoperatively to 6 months postoperatively in TKA patients. The main predictors associated with the FJS at 6 months after surgery were sex, combined MSD, operation time, FJS (T3), ROM (T6), pain score (T6), GO-SSS score (T6), and anxiety (T6).
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Artroplastia do Joelho , Osteoartrite do Joelho , Humanos , Artroplastia do Joelho/métodos , Modelos Lineares , Estudos Prospectivos , Estudos Longitudinais , Osteoartrite do Joelho/cirurgia , Articulação do Joelho/cirurgia , Dor , Estudos RetrospectivosRESUMO
INTRODUCTION: Orthopaedic trauma patients are at high risk of venous thromboembolism (VTE). As VTE prophylaxis has gradually raised public concerns, guidelines related to this topic have increased over time. However, the existing recommendations of thromboprophylaxis guidelines in orthopaedic trauma patients are still inconsistent, and the quality of the guidelines and recommendations for the topic still lacks comprehensive assessments. This review aims to critically appraise clinical practice guidelines for thromboprophylaxis in orthopaedic trauma patients. METHODS AND ANALYSIS: We will conduct a comprehensive literature search up to 31 October 2022 in databases (PubMed, EMBASE, CINAHL, Web of Science, the Cochrane Library, etc), academic websites and guideline repositories. The quality of the guidelines and recommendations will be assessed by five reviewers independently using the Appraisal of Guidelines Research and Evaluation II instrument (AGREE-II) and the AGREE - Recommendation EXcellence (AGREE-REX). We will summarise the characteristics of the guidelines and compare the differences between these recommendations. ETHICS AND DISSEMINATION: This study will follow the Declaration of Helsinki and has received approval from the Ethics Committee on Biomedical Research, West China Hospital, Sichuan University (ethics approval no. 2021-989). The results will be summarised as a paper, disseminated through peer-reviewed journals, and will help guide further research in the future. PROTOCOL REGISTRATION NUMBER: CRD42021273405.
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Ortopedia , Tromboembolia Venosa , Humanos , Anticoagulantes/uso terapêutico , China , Bases de Dados Factuais , Literatura de Revisão como Assunto , Tromboembolia Venosa/prevenção & controle , Revisões Sistemáticas como AssuntoRESUMO
Nuclear factor κB (NF-κB) is involved in a wide range of innate immune activities in host cells and serves as an important component of a host's immunity system. To survive in infected cells, viruses have evolved intricate strategies to evade the host immune response. Pseudorabies virus (PRV) is a member of the alpha herpesvirus family and is capable of causing reproductive and neurological dysfunction in pigs. PRV has a large DNA genome and therefore has the ability to encode numerous proteins that modulate host innate immune responses. In the present study, we demonstrated that the PRV-encoded immediate early protein ICP0 inhibits the tumor necrosis factor alpha (TNF-α)-mediated NF-κB signaling pathway. An in-depth study showed that ICP0 protein was able to limit NF-κB activation and decreased the expression of inflammatory cytokines interleukin-6 (IL-6) and interleukin 8 (IL-8). In addition, ICP0 blocked the activation of NF-κB through interacting with p65, degrading its protein expression and limiting its phosphorylation. PRV protein ICP0 is shown for the first time to enable escape from innate immune response through the regulation of NF-κB during PRV infection. These results illustrate that PRV ICP0 is able to block NF-κB activation. This mechanism may represent a critical role in the early events leading to PRV infection.
Assuntos
Herpesvirus Suídeo 1 , Proteínas Imediatamente Precoces , Animais , Linhagem Celular , Herpesvirus Suídeo 1/metabolismo , Proteínas Imediatamente Precoces/genética , NF-kappa B/metabolismo , Suínos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Heat shock proteins (HSPs) are a protein family that respond to physiological stress, such as heat, starvation, and infection. As cellular protein chaperones, they play an important role in protein folding, assembly, and degradation. Though it is well known that HSP27 is involved in a range of viral infections, its role during an encephalomyocarditis virus (EMCV) infection is not known. Here, we report that EMCV degrades HSP27 and that EMCV proteins 2Cpro and 3Apro are primarily responsible for its degradation. Consequently, loss of cellular HSP27 augmented EMCV proliferation, an effect that could be reversed upon HSP27 overexpression. Importantly, we found that HSP27 positively regulated EMCV-triggered type I interferon (IFN) production. Moreover, overexpression of 2Cpro and 3Apro significantly blocked type I IFN production. We also found for the first time that HSP27, as a molecular chaperone, can specifically interact with MDA5 and stabilize the expression of MDA5. Collectively, this study shows that HSP27 dampens EMCV infectivity by positively regulating EMCV-triggered retinoic acid-inducible gene (RIG)-I-like receptor (RLR)/melanoma differentiation-associated gene 5 (MDA5) signal pathway, while EMCV proteins 2Cpro and 3Apro interact with HSP27 and degrade HSP27 protein expression to allow EMCV proliferation. Our findings provide further mechanistic evidence for EMCV partaking in immune escape mechanisms, and that 2Cpro and 3Apro could serve as potential antiviral targets.
RESUMO
Type I interferon (IFN)-mediated antiviral responses are critical for modulating host-virus responses, and indeed, viruses have evolved strategies to antagonize this pathway. Encephalomyocarditis virus (EMCV) is an important zoonotic pathogen, which causes myocarditis, encephalitis, neurological disease, reproductive disorders, and diabetes in pigs. This study aims to understand how EMCV interacts with the IFN pathway. EMCV circumvents the type I IFN response by expressing proteins that antagonize cellular innate immunity. Here, we show that EMCV VP2 is a negative regulator of the IFN-ß pathway. This occurs via the degradation of the MDA5-mediated cytoplasmic double-stranded RNA (dsRNA) antiviral sensing RIG-I-like receptor (RLR) pathway. We show that structural protein VP2 of EMCV interacts with MDA5, MAVS, and TBK1 through its C terminus. In addition, we found that EMCV VP2 could significantly degrade RLRs by the proteasomal and lysosomal pathways. For the first time, EMCV VP2 was shown to play an important role in EMCV evasion of the type I IFN signaling pathway. This study expands our understanding that EMCV utilizes its capsid protein VP2 to evade the host antiviral response.IMPORTANCE Encephalomyocarditis virus is an important pathogen that can cause encephalitis, myocarditis, neurological diseases, and reproductive disorders. It also causes huge economic losses for the swine industry worldwide. Innate immunity plays an important role in defending the host from pathogen infection. Understanding pathogen microorganisms evading the host immune system is of great importance. Currently, whether EMCV evades cytosolic RNA sensing and signaling is still poorly understood. In the present study, we found that viral protein VP2 antagonized the RLR signaling pathway by degrading MDA5, MAVS, and TBK1 protein expression to facilitate viral replication in HEK293 cells. The findings in this study identify a new mechanism for EMCV evading the host's innate immune response, which provide new insights into the virus-host interaction and help develop new antiviral approaches against EMCV.
Assuntos
Proteínas do Capsídeo/metabolismo , Vírus da Encefalomiocardite/fisiologia , Interferon beta/metabolismo , Transdução de Sinais , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas do Capsídeo/química , Proteínas do Capsídeo/genética , Infecções por Cardiovirus/imunologia , Infecções por Cardiovirus/virologia , Proteína DEAD-box 58/antagonistas & inibidores , Proteína DEAD-box 58/metabolismo , Vírus da Encefalomiocardite/genética , Vírus da Encefalomiocardite/metabolismo , Células HEK293 , Humanos , Evasão da Resposta Imune , Imunidade Inata , Interferon Tipo I/metabolismo , Helicase IFIH1 Induzida por Interferon/metabolismo , Mutação , Domínios e Motivos de Interação entre Proteínas , Proteínas Serina-Treonina Quinases/metabolismo , Receptores Imunológicos/antagonistas & inibidores , Receptores Imunológicos/metabolismo , Replicação ViralRESUMO
OBJECTIVE: To examine whether a modified endometriosis fertility index (EFI) can better predict the rate of pregnancy without assisted reproductive technologies (ART) after laparoscopic surgery in infertile Chinese women with endometriosis. METHODS: 564 infertile women undergoing laparoscopy for endometriosis were retrospectively collected from January 2014 to December 2018. 472 patients were used to modify the EFI based on new, optimal cutoffs for its predictor variables. The predictive accuracy of the modified EFI was examined in the other 92 patients. RESULTS: Among the patients for the EFI modification, the multivariable Cox regression results showed that historical factors made more contribution in predicting non-ART pregnancy rate than surgical factors both in modified EFI (C-index: historical factors 0.617 vs surgical factors 0.558) and original EFI (C-index: historical factors 0.600 vs surgical factors 0.549). No significant relationship between the prior pregnancy and post-operative non-ART pregnancy rates was detected by both modified EFI and original EFI (p = 0.530 and 0.802, respectively). To assess the predictive effect of modified EFI, the two versions of modified EFI not only had higher predictive accuracy (C-index: 0.627 and 0.632) for non-ART pregnancy rates than that of the original EFI (C-index: 0.602) in the patients undergoing surgery during 2014-2017, but also higher than that of the original EFI (C-index: 0.638 and 0.612 vs 0.560) in the externally validated population in 2018. CONCLUSIONS: A modified EFI based on population-specific optimal cutoffs and weights might be more suitable for estimating the rate of non-ART pregnancy after laparoscopic surgery in infertile women with endometriosis.
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Endometriose/cirurgia , Fertilidade/fisiologia , Taxa de Gravidez/tendências , Adulto , China , Estudos de Coortes , Feminino , Humanos , GravidezRESUMO
At present, the 2019-nCoV epidemic situation is in severe and complex period. In order to prevent the virus from invading and infecting, it is very important and urgent for medical personnel to protect themselves. However, in the process of using protective equipment by medical personnel, the performance of device related pressure injuries (DRPI) caused by pain, numbness, redness, and even breakage caused by the equipment has seriously endangered the health of medical personnel. This article, based on Prevention and Treatment of Pressure Ulcers/Injuries: Quick Reference Guide 2019, references, and clinical experiences of wound specialists in West China Hospital of Sichuan University, summarize the preventive and protective measures of West China Hospital for medical personnel to prevent DRPI, so as to provide clinical preventive measurements for medical personnel.
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COVID-19 , Equipamento de Proteção Individual , Recursos Humanos em Hospital , Úlcera por Pressão , Humanos , China/epidemiologia , COVID-19/epidemiologia , COVID-19/prevenção & controle , Equipamento de Proteção Individual/efeitos adversos , Úlcera por Pressão/epidemiologia , Úlcera por Pressão/prevenção & controle , EpidemiasRESUMO
Manganese oxides are promising anode materials for their high-energy density. However, they suffer from poor rate capability and fast capacity fading. Herein, we construct a three-dimensional (3D) core-shell structured polypyrrole (PPy)/MnO2-reduced graphene oxide (rGO)-carbon nanotubes (CNTs) composite via a facile two-step method. In the structure, the CNTs can facilitate fast electron conduction and keep structural integrity. The flexible and conductive rGO nanosheets work as both a reactive material and a carrier for MnO2 in-situ growth. The MnO2 nanosheets well distributed on the rGO/CNTs scaffold favor the energy storage by way of fast Li+ insertion and extraction. PPy nanoparticles (â¼10nm) well wrapped on the MnO2 nanosheets not only enable the interfacial stabilization, but also provide a buffer layer to accommodate the volume expansion. As a result, the as-prepared PPy/MnO2-rGO-CNTs composite exhibits high specific capacity, excellent cycling stability and good rate capability. A reversible specific capacity of 1748.1mAhg-1 is achieved at the current density of 100mAg-1 after 200 cycles. Even at a high current density of 1000mAg-1, the composite still exhibits 941.1mAhg-1 after 1200 cycles. The design strategy of the composite can be extended to other high-capacity metal oxide material.
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OBJECTIVE: To explore the efficacy of laparoscopic surgery without auxiliary treatment for type II cesarean scar pregnancy (CSP-II). STUDY DESIGN: This was a case series of 7 patients with CSP-II who underwent laparoscopic surgery without auxiliary treatment between April 2014 and April 2015. All cases were diagnosed by ultrasound, confirmed by laparoscopy, and managed by laparoscopic resection of scar and gestational tissue and wound repair. RESULTS: All 7 patients had successful surgeries without complication. Uterine scar and gestational tissues were resected, while also preserving the uterus. The operation time was 70.1 ± 16.3 min and blood loss was 65.7 ± 32.1 mL. Serum ß-hCG levels 24 hours after surgery declined by 84.8 ± 9.4%. Serum ß-hCG levels went back to <5 IU/L in all 7 patients by 14.4 ± 4.3 days after surgery. The time interval between surgery and first menstruation was 35.3 ± 4.5 days. CONCLUSION: These results suggest the possibility that skilled surgeons could use laparoscopy without auxiliary pretreatment to remove gestational tissues and uterine scar defect and to repair the wound in patients with CSP-II.
Assuntos
Cesárea/efeitos adversos , Cicatriz/cirurgia , Laparoscopia , Gravidez Ectópica/cirurgia , Adulto , Perda Sanguínea Cirúrgica , Gonadotropina Coriônica Humana Subunidade beta/sangue , Cicatriz/classificação , Feminino , Humanos , Duração da Cirurgia , Gravidez , Gravidez Ectópica/diagnóstico por imagem , UltrassonografiaRESUMO
Warburg effect is characterized by an increased utilization of glucose via glycolysis in cancer cells, even when enough oxygen is present to properly respire. Recent studies demonstrate that deregulation of microRNAs contributes to the Warburg effect. In the present study, we show that miR-144 is downregulated while glucose transporter 1 (Glut1) is upregulated in ovarian cancers. In vitro studies further showed that miR-144 inhibits Glut1 expression through targeting its 3'-untranslated region. As a result, cells overexpressing miR-144 exhibited a metabolic shift, including enhanced glucose uptake and lactate production. The altered glucose metabolism induced by miR-144 also leads to a rapid growth of ovarian cancer cells. Taken together, our results indicate that miR-144 may serve as a molecular switch to regulate glycolysis in ovarian cancer by targeting the expression of Glut1.