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BACKGROUND Osteoarthritis (OA) is a chronic degenerative disease characterized by synovitis and has been implicated in sphingolipid metabolism disorder. However, the role of sphingolipid metabolism pathway (SMP)-related genes in the occurrence of OA and synovial immune dysregulation remains unclear. MATERIAL AND METHODS In this study, we obtained synovium-related databases from GEO (n=40 for both healthy controls and OA) and analyzed the expression levels of SMP-related genes. Using 2 algorithms, we identified hub genes and developed a diagnostic model incorporating these hub genes to predict the occurrence of OA. Subsequently, the hub genes were further validated in peripheral blood samples from OA patients. Additionally, CIBERSORT and MCP-counter analyses were employed to explore the correlation between hub genes and immune dysregulation in OA synovium. WGCNA was used to determine enriched modules in different clusters. RESULTS Overall, the expression levels of SMP genes were upregulated in OA synovium. We identified 6 hub genes of SMP and constructed an excellent diagnostic model (AUC=0.976). The expression of re-confirmed hub genes showed associations with immune-related cell infiltration and levels of inflammatory cytokines. Furthermore, we observed heterogeneity in the expression patterns of hub genes across different clusters of OA. Notably, older patients displayed increased susceptibility to elevated levels of pain-related inflammatory cytokines and infiltration of immune cells. CONCLUSIONS The SMP-related hub genes have the potential to serve as diagnostic markers for OA patients. Moreover, the 4 hub genes of SMP demonstrate wide participation in immune dysregulation in OA synovium. The activation of different pathways is observed among different populations of patients with OA.
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Osteoartritis , Esfingolípidos , Membrana Sinovial , Humanos , Membrana Sinovial/metabolismo , Osteoartritis/genética , Osteoartritis/diagnóstico , Osteoartritis/metabolismo , Osteoartritis/inmunología , Esfingolípidos/metabolismo , Perfilación de la Expresión Génica/métodos , Redes Reguladoras de Genes , Masculino , Femenino , Transcriptoma/genética , Bases de Datos Genéticas , Persona de Mediana Edad , Estudios de Casos y ControlesRESUMEN
Osteoarthritis (OA) is a prevalent joint disease that can be exacerbated by lipid metabolism disorders. The intra-articular fat pad (IFP) has emerged as an active participant in the pathological changes of knee OA (KOA). However, the proteomic and lipidomic differences between IFP tissues from KOA and control individuals remain unclear. Samples of IFP were collected from individuals with and without OA (n = 6, n = 6). Subsequently, these samples underwent liquid chromatography/mass spectrometry-based label-free quantitative proteomic and lipidomic analysis to identify differentially expressed proteins (DEPs) and lipid metabolites (DELMs). The DEPs were further subjected to enrichment analysis, and hub DEPs were identified using multiple algorithms. Additionally, an OA diagnostic model was constructed based on the identified hub DEPs or DELMs. Furthermore, CIBERSORT was utilized to investigate the correlation between hub protein expression and immune-related modules in IFP of OA. Our results revealed the presence of 315 DEPs and eight DELMs in IFP of OA patients compared to the control group. Enrichment analysis of DEPs highlighted potential alterations in pathways related to coagulation, complement, fatty acid metabolism, and adipogenesis. The diagnostic model incorporating four hub DEPs (AUC = 0.861) or eight DELMs (AUC = 0.917) exhibited excellent clinical validity for diagnosing OA. Furthermore, the hub DEPs were found to be associated with immune dysfunction in IFP of OA. This study presents a distinct proteomic and lipidomic landscape of IFP between individuals with OA and those without. These findings provide valuable insights into the molecular changes associated with potential mechanisms underlying OA.
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Tejido Adiposo , Lipidómica , Osteoartritis de la Rodilla , Proteómica , Humanos , Osteoartritis de la Rodilla/metabolismo , Osteoartritis de la Rodilla/patología , Tejido Adiposo/metabolismo , Tejido Adiposo/patología , Proteómica/métodos , Lipidómica/métodos , Masculino , Femenino , Persona de Mediana Edad , Metabolismo de los Lípidos , Anciano , Relevancia ClínicaRESUMEN
Rotator cuff tears are still among the serious challenges encountered by sports medicine surgeons. With the development of arthroscopic technology, the treatment options for irreparable massive rotator cuff tears (IMRCTs) have gradually increased, but postoperatively, these options are prone to graft retears, reoperations, infections, revision surgical procedures, and so on. On the basis of studies on the balance of the force couples of the rotator cuff, more scholars believe that when addressing IMRCTs, attention should be paid to the reconstruction of the force couples of the rotator cuff; hence, we developed the superior fulcrum reconstruction technique. This article describes an alternative approach to IMRCT repair with superior fulcrum reconstruction using autologous ipsilateral peroneus longus tendon.
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Objective: To investigate early effectiveness of arthroscopic superior fulcrum reconstruction in the treatment of irreparable massive rotator cuff tear (IMRCT). Methods: A retrospective analysis was conducted on the clinical data of 24 patients with IMRCT who met the inclusion criteria between January 2020 and April 2022. Among them, there were 11 males and 13 females with an average age of 56.2 years (range, 42-68 years). There were 12 cases of falling injuries, 3 cases of traction injuries, and the other 9 cases had no obvious causes. The disease duration ranged from 1 to 25 months (median, 6 months). The rotator cuff tears were classified as Hamada grade 2 in 18 cases and grade 3 in 6 cases, and Goutallier grade 1 in 3 cases, grade 2 in 20 cases, and grade 3 in 1 case. All patients were treated with arthroscopic superior fulcrum reconstruction. Visual analogue scale (VAS) score, Constant-Murley score, the University of California at Los Angeles (UCLA) score, and the American Shoulder and Elbow Surgeons (ASES) score were recorded before operation and at 1, 3, 6, and 12 months after operation. Results: The operations were all successfully completed. The incisions healed by first intention and no related complications occurred. All patients were followed up 12-33 months (mean, 24.6 months). The VAS, Constant-Murley, UCLA, and ASES scores at different time points after operation were superior to those before operation ( P<0.05). All of the above indicators further improved with time. Except for no significant difference in VAS and Constant-Murley scores between 6 and 12 months ( P>0.05), the differences between the other time points were significant ( P<0.05). At 12 months after operation, according to UCLA scoring standard, shoulder joint function was rated as excellent in 4 cases, good in 19 cases, and poor in 1 case, with an excellent and good rate of 96.0%. MRI showed that there was no graft re-tear and the transplanted tendon and bone tunnel healed. Conclusion: The arthroscopic superior fulcrum reconstruction for IMRCT can effectively relieve the pain, improve the shoulder range of motion, and restore good shoulder function.
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Lesiones del Manguito de los Rotadores , Articulación del Hombro , Masculino , Femenino , Humanos , Persona de Mediana Edad , Lesiones del Manguito de los Rotadores/cirugía , Manguito de los Rotadores/cirugía , Estudios Retrospectivos , Resultado del Tratamiento , Artroscopía , Articulación del Hombro/cirugía , Rango del Movimiento ArticularRESUMEN
BACKGROUND: Schatzker IV-C is a high-energy tibial plateau fracture often accompanied by lateral meniscus injuries. While imaging examinations are routine preoperative measurements, the correlation between CT imaging shift parameters of the lateral plateau and lateral meniscal injury in Schatzker IV-C fractures remains uncovered. METHODS: This retrospective study enrolled a total of 60 patients with Schatzker IV-C tibial plateau fractures at the First People's Hospital of Hefei. Prior to surgery, CT imaging was used to measure the numerical values of lateral plateau depression (LPD) and lateral plateau widening (LPW). The degree of lateral meniscus injury was confirmed based on intraoperative direct vision, with patients being classified into meniscus injury and non-meniscus injury groups. Dichotomous logistic regression was employed to evaluate the correlation between LPD, LPW, and lateral meniscus injury, while the optimal cut-off points for predicting lateral meniscal injury with LPD and LPW were determined using receiver operator characteristic (ROC) curves. RESULTS: The meniscus injury group exhibited a mean LPD of 15.3 ± 3.5 mm, which was significantly higher than the non-meniscus injury group's mean LPD of 8.4 ± 3.4 mm (P < 0.05). Similarly, the meniscus injury group had a larger mean LPW of 9.4 ± 1.8 mm compared to the non-meniscus injury group's mean LPW of 6.9 ± 0.9 mm (P < 0.05). The optimal cut-off points for predicting lateral meniscal injury were determined to be 8.40 mm for LPD (with a sensitivity of 95%, specificity of 85%, and AUC of 0.898) and 7.90 mm for LPW (with a sensitivity of 75%, specificity of 90%, and AUC of 0.897). CONCLUSIONS: Patients with Schatzker IV-C tibial plateau fractures are at a significantly higher risk of lateral meniscal injury when the LPD exceeds 8.40 mm and/or the LPW exceeds 7.90 mm. Our results may provide novel reference metrics for the early diagnosis of lateral meniscal injury in Schatzker IV-C tibial plateau fracture patients when the MRI examination is not available.
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Fracturas de la Tibia , Fracturas de la Meseta Tibial , Humanos , Meniscos Tibiales/diagnóstico por imagen , Meniscos Tibiales/cirugía , Estudios Retrospectivos , Fracturas de la Tibia/diagnóstico por imagen , Fracturas de la Tibia/cirugía , Fracturas de la Tibia/complicaciones , Tomografía Computarizada por Rayos X/métodosRESUMEN
Circular RNA (circRNA) THBS1 has been shown to exist as an oncogene in non-small-cell lung cancer, but its role in cervical cancer is still unclear. Our experiment aimed to uncover the functions and specific mechanism of circRNA THBS1 in cervical cancer cells. Levels of circRNA THBS1 and miR-543 in cervical cancer tissues and cell lines were assessed by RT-qPCR. starBase and dual luciferase reporter gene assay were applied for investigating the correlation between miR-543 and circRNA THBS1/HMGB2. Cell proliferation and apoptosis were evaluated by MTT and flow cytometry, respectively. Furthermore, the levels of HMGB2, E-cadherin, and N-cadherin in HeLa cells were determined by RT-qPCR and western blot analysis. Our data revealed that circRNA THBS1 was significantly upregulated and miR-543 was low expressed in cervical cancer tissues and cell lines. circRNA THBS1 interacted with miR-543 and negatively regulated miR-543 expression in HeLa cells. Silencing of circRNA THBS1 remarkably suppressed HeLa cells' viability, accelerated cells' apoptosis, and inhibited the EMT of HeLa cells, while these changes were reversed by miR-543 inhibitor. Moreover, miR-543 affected HeLa cells by targeting HMGB2. In conclusion, circRNA THBS1 silencing inhibited the malignant biological behaviors of cervical cancer cells via the regulation of miR-543/HMGB2 axis.
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Due to the large specific surface area and rich pore structure, chitosan nanofiber membrane has many advantages over conventional gel-like or film-like products. However, the poor stability in acidic solutions and relatively weak antibacterial activity against Gram-negative bacteria severely restrict its use in many industries. Here, we present a chitosan-urushiol composite nanofiber membrane prepared by electrospinning. Chemical and morphology characterization revealed that the formation of chitosan-urushiol composite involved the Schiff base reaction between catechol and amine groups and the self-polymerization of urushiol. The unique crosslinked structure and multiple antibacterial mechanisms endowed the chitosan-urushiol membrane with outstanding acid resistance and antibacterial performance. After immersion in HCl solution at pH 1, the membrane maintained its intact appearance and satisfactory mechanical strength. In addition to its good antibacterial performance against Gram-positive Staphylococcus aureus (S. aureus), the chitosan-urushiol membrane exhibited synergistic antibacterial activity against Gram-negative Escherichia coli (E. coli) that far exceeded that of neat chitosan membrane and urushiol. Moreover, cytotoxicity and hemolysis assays revealed that the composite membrane had good biocompatibility similar to that of neat chitosan. In short, this work provides a convenient, safe, and environmentally friendly method to simultaneously enhance the acid resistance and broad-spectrum antibacterial activity of chitosan nanofiber membranes.
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Quitosano , Nanofibras , Quitosano/farmacología , Quitosano/química , Nanofibras/química , Staphylococcus aureus , Escherichia coli , Antibacterianos/farmacología , Antibacterianos/química , CatecolesRESUMEN
The cGAS-STING pathway mediates cytoplasmic DNA-triggered innate immunity. STING activation is initiated by cyclic-GMP-AMP (cGAMP)-induced translocation from the endoplasmic reticulum and sulfated glycosaminoglycans-induced polymerization at the Golgi. Here, we examine the mechanisms underlying STING transport and activation beyond the Golgi. A genome-wide CRISPR-Cas9 screen identified Armadillo-like helical domain-containing protein 3 (ARMH3) as critical for STING activation. Upon cGAMP-triggered translocation, ARMH3 interacted with STING at the Golgi and recruited phosphatidylinositol 4-kinase beta (PI4KB) to synthesize PI4P, which directed STING Golgi-to-endosome trafficking via PI4P-binding proteins AP-1 and GGA2. Disrupting PI4P-dependent lipid transport through RNAi of other PI4P-binding proteins impaired STING activation. Consistently, disturbed lipid composition inhibited STING activation, whereas aberrantly elevated cellular PI4P led to cGAS-independent STING activation. Armh3fl/fllLyzCre/Cre mice were susceptible to DNA virus challenge in vivo. Thus, ARMH3 bridges STING and PIK4B to generate PI4P for STING transportation and activation, an interaction conserved in all eukaryotes.
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Factores de Restricción Antivirales , Proteínas del Dominio Armadillo , Proteínas de la Membrana , Animales , Ratones , 1-Fosfatidilinositol 4-Quinasa/metabolismo , Proteínas Portadoras , Endosomas/metabolismo , Inmunidad Innata , Lípidos , Proteínas de la Membrana/metabolismo , Nucleotidiltransferasas/metabolismo , Proteínas del Dominio Armadillo/metabolismoRESUMEN
BACKGROUND: Osteoarthritis (OA) is one of degenerative-related arthritis, which can be aggravated by low-grade synovitis. It is known that arachidonic acid (AA) dysmetabolism brings OA synovitis. However, the impact of synovial AA metabolism pathway (AMP) related genes on OA remains uncovered. METHODS: Here, we conducted a comprehensive analysis to explore the impact of AA metabolism genes in OA synovium. We obtained transcriptome expression profiles from three raw datasets related to OA synovium (GSE12021, GSE29746, GSE55235) and identified the hub genes of AA metabolism pathways (AMP) in OA synovium. An OA occurrence diagnostic model was constructed and validated based on the identified hub genes. Then, we explored the correlation between hub gene expression and the immune-related module using CIBERSORT and MCP-counter analysis. The unsupervised consensus clustering analysis and weighted correlation network analysis (WGCNA) were utilized to identify robust clusters of identified genes in each cohort. Moreover, the interaction between the hub genes of AMP and immune cells was elucidated through single-cell RNA (scRNA) analysis by scRNA sequencing data from GSE152815. RESULTS: We found that the expression of AMP-related genes was up-regulated in OA synovium, and seven hub genes (LTC4S, PTGS2, PTGS1, MAPKAPK2, CBR1, PTGDS, and CYP2U1) were identified. The diagnostic model that combined the identified hub genes showed great clinical validity in diagnosing OA (AUC = 0.979). Moreover, significant associations were noticed between the hub genes' expression, immune cell infiltration, and inflammatory cytokine levels. The 30 OA patients were randomized and clustered into three groups using WGCNA analysis based on the hub genes, and diverse immune status was found in different clusters. Of interest, older patients were more likely to be classified into a cluster with higher levels of inflammatory cytokines IL-6 and less infiltration of immune cells. Based on the scRNA-sequencing data, we found that the hub genes had relatively higher expression in macrophages and B cells than other immune cells. Moreover, inflammation-related pathways were significantly enriched in macrophages. CONCLUSION: These results suggest that AMP-related genes are closely involved in alterations of OA synovial inflammation. The transcriptional level of hub genes could serve as a potential diagnostic marker for OA.
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Metabolismo de los Lípidos , Osteoartritis , Humanos , Ácido Araquidónico , Citocinas , Inflamación , Osteoartritis/diagnóstico , Osteoartritis/genética , Análisis de Secuencia de ARN , Familia 2 del Citocromo P450RESUMEN
PURPOSE: Postoperative adjuvant trans-catheter arterial chemoembolization (TACE) is regarded as a common strategy for hepatocellular carcinoma (HCC) patients at a high risk of recurrence. However, there are currently no clinically available biomarkers to predict adjuvant TACE response. Vessels that encapsulate tumor clusters (VETC) can be used as an independent predictor of HCC prognosis. In this study, we aimed to explore whether the VETC pattern could predict adjuvant TACE benefit. METHODS: Vascular pattern and HIF-1α expression were detected in immunohistochemistry. The survival benefit of adjuvant TACE therapy for patients with or without VETC pattern (VETC+ /VETC-) was evaluated. RESULTS: The adjuvant TACE therapy obviously improved the TTR and OS in VETC+ patients, while adjuvant TACE therapy could not benefit from VETC- patients. Univariate and multivariate analysis revealed that adjuvant TACE therapy significantly improved the TTR and OS in VETC+ patients, but not in VETC- patients. In addition, the VETC+ , but not VETC- , patients could benefit from adjuvant TACE therapy in patients with high-risk factors of vascular invasion, larger tumor or multiple tumor. The mechanistic investigations revealed that the favorable efficacy of adjuvant TACE on VETC+ patients, but not VETC- ones, may be not due to the activation of HIF-1α pathway. CONCLUSION: The VETC pattern may represent a novel and reliable factor for selecting HCC patients who may benefit from adjuvant TACE therapy, and the combination of VETC pattern and tumor characteristics may help stratify patients' outcomes and responses to adjuvant TACE therapy.
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Carcinoma Hepatocelular , Quimioembolización Terapéutica , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/patología , Neoplasias Hepáticas/patología , Pronóstico , Análisis Multivariante , Terapia Combinada , Estudios RetrospectivosRESUMEN
The cGAS-STING pathway is responsible for cytoplasmic double-stranded DNA (dsDNA) -triggered innate immunity and involved in the pathology of various diseases including infection, autoimmune diseases, neurodegeneration and cancer. Understanding the activation and regulatory mechanisms of this pathway is critical to develop therapeutic strategies toward these diseases. Here, we review the signal transduction, cellular functions and regulations of cGAS and STING, particularly highlighting the latest understandings on the activation of cGAS by dsDNA and/or Manganese (Mn2+), STING trafficking, sulfated glycosaminoglycans (sGAGs)-induced STING polymerization and activation, and also regulation of the cGAS-STING pathway by different biocondensates formed via phase separation of proteins from host cells and viruses.
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Enfermedades Autoinmunes , Proteínas de la Membrana , Humanos , Animales , Proteínas de la Membrana/metabolismo , Nucleotidiltransferasas/metabolismo , Inmunidad Innata , Transducción de SeñalRESUMEN
Dried blood spot (DBS) based PCR was considered an inexpensive and feasible method for detecting pathogens in the blood. The DBS carrier filter paper and PCR kits are crucial for accurate diagnosis. We evaluated 4 types of filter papers and 20 PCR kits for DBS samples. The PCR detecting Plasmodium results showed that the minimum detection limit of the 4 filter papers was 1 × 102 parasites/µL, and the positive rates of 20 PCR kits ranged from 0% to 100%. PCR results were satisfactory for detecting Plasmodium falciparum (P. falciparum) and Plasmodium. vivax (P. vivax) in archived DBS samples and Babesia gibsoni (B. gibsoni) in fresh pet DBS samples. Our results provided a useful reference for the detection of blood pathogens with DBS samples and direct PCR, especially for screening the cost-efficacy combination of filter paper and PCR kit in resource-limited areas.
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Solvent-free mechanical milling is a new, environmentally friendly and cost-effective technology that is now widely used in the field of organic synthesis. The mechanochemical solvent-free synthesis of furoxans from aldoximes was achieved through dimerization of the in situ generated nitrile oxides in the presence of sodium chloride, Oxone and a base. A variety of furoxans was obtained with up to a 92% yield. The present protocol has the advantages of high reaction efficiency and mild reaction conditions.
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Oxadiazoles , Oximas , Técnicas de Química Sintética , Dimerización , SolventesRESUMEN
Programmed death ligand-1 (PD-L1)/PD-1 checkpoint extensively serves as a central mediator of immunosuppression. A tumor-promoting role for abundant PD-L1 in several cancers is revealed. However, the importance of PD-L1 and how the PD-L1 expression is controlled in breast cancer remains obscure. Here, the mechanisms of controlling PD-L1 at the transcription and protein acetylation levels in promoting breast cancer growth are presented. Overexpressed PD-L1 accelerates breast cancer growth in vitro and in vivo. RNA-seq uncovers that PD-L1 can induce some target genes affecting many cellular processes, especially cancer development. In clinical breast cancer tissues and cells, PD-L1 and HBXIP are both increased, and their expressions are positively correlated. Mechanistic exploration identifies that HBXIP stimulates the transcription of PD-L1 through co-activating ETS2. Specifically, HBXIP induces PD-L1 acetylation at K270 site through interacting with acetyltransferase p300, leading to the stability of PD-L1 protein. Functionally, depletion of HBXIP attenuates PD-L1-accelerated breast tumor growth. Aspirin alleviates breast cancer via targeting PD-L1 and HBXIP. Collectively, the findings display new light into the mechanisms of controlling tumor PD-L1 and broaden the utility for PD-L1 as a target in breast cancer therapy.
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Proteínas Adaptadoras Transductoras de Señales/metabolismo , Antígeno B7-H1/metabolismo , Neoplasias de la Mama/patología , Animales , Western Blotting , Neoplasias de la Mama/metabolismo , Línea Celular Tumoral , Proliferación Celular , Inmunoprecipitación de Cromatina , Femenino , Técnica del Anticuerpo Fluorescente , Humanos , Células MCF-7 , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
BACKGROUND: Diosmetin is a bioflavonoid compound naturally abundant in citrus fruits. It is found to perform a variety of activities, while its antitumor property in osteosarcoma, a malignant tumor with unmet clinical treatment, remained unknown. METHODS: Colony formation assay, cell cycle analysis and apoptosis analysis were conducted respectively to observe the effect of diosmetin on cell proliferation and apoptosis in human osteosarcoma cells. Western blot and immunoprecipitation were used to detect the expression of apoptotic molecules and activation of STAT3/c-Myc pathway in Saos-2 and U2SO cells. RESULTS: Diosmetin significantly inhibited cell proliferation, induced cell cycle arrest at G2/M phase and promoted cell apoptosis in both Saos-2 and U2SO cells. Moreover, Diosmetin downregulated the expression of anti-apoptotic protein Bcl-xL while upregulated the levels of pro-apoptotic proteins including cleaved Caspase-3, cleaved-PARP and Bax. Furthermore, diosmetin dose-dependently inhibited STAT3 phosphorylation, reduced the expression of its downstream protein c-Myc and impeded the interaction between STAT3 molecules. CONCLUSIONS: These results suggest that diosmetin exerts anti-osteosarcoma effects by suppressing cell proliferation and inducing apoptosis via inhibiting the activation of STAT3/c-Myc signaling pathway, which provide the possibility for diosmetin to be a chemotherapeutic candidate for osteosarcoma.
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Flavonoides , Osteosarcoma , Transducción de Señal/efectos de los fármacos , Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Flavonoides/farmacología , Humanos , Osteosarcoma/tratamiento farmacológico , Proteínas Proto-Oncogénicas c-myc , Factor de Transcripción STAT3RESUMEN
Biomolecular condensates (biocondensates) formed via liquid-liquid phase-separation of soluble proteins have been studied extensively. However, neither the phase-separation of endoplasmic reticulum (ER) transmembrane protein nor a biocondensate with organized membranous structures has been reported. Here, we have discovered a spherical ER membranous biocondensate with puzzle-like structures caused by condensation of the ER-resident stimulator of interferon genes (STING) in DNA virus-infected or 2'3'-cGAMP (cyclic GMP-AMP)-treated cells, which required STING transmembrane domains, an intrinsically disordered region (IDR) and a dimerization domain. Intracellular 2'3'-cGAMP concentrations determined STING translocation or condensation. STING biocondensates constrained STING and TBK1 (TANK binding protein 1) to prevent innate immunity from overactivation, presumably acting like a 'STING-TBK1-cGAMP sponge'. Cells expressing STING-E336G/E337G showed notably enhanced innate immune responses due to impaired STING condensation after viral infection at later stages. Microtubule inhibitors impeded the STING condensate gel-like transition and augmented type I-interferon production in DNA virus-infected cells. This membranous biocondensate was therefore named the STING phase-separator.
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Retículo Endoplásmico/genética , Inmunidad Innata/genética , Extracción Líquido-Líquido , Proteínas de la Membrana/genética , Humanos , Nucleótidos Cíclicos/genética , Unión Proteica/genética , Proteínas Serina-Treonina Quinasas/genética , Transducción de Señal/genéticaRESUMEN
Activation of the cyclic guanosine monophosphate (GMP)-AMP (cGAMP) sensor STING requires its translocation from the endoplasmic reticulum to the Golgi apparatus and subsequent polymerization. Using a genome-wide CRISPR-Cas9 screen to define factors critical for STING activation in cells, we identified proteins critical for biosynthesis of sulfated glycosaminoglycans (sGAGs) in the Golgi apparatus. Binding of sGAGs promoted STING polymerization through luminal, positively charged, polar residues. These residues are evolutionarily conserved, and selective mutation of specific residues inhibited STING activation. Purified or chemically synthesized sGAGs induced STING polymerization and activation of the kinase TBK1. The chain length and O-linked sulfation of sGAGs directly affected the level of STING polymerization and, therefore, its activation. Reducing the expression of Slc35b2 to inhibit GAG sulfation in mice impaired responses to vaccinia virus infection. Thus, sGAGs in the Golgi apparatus are necessary and sufficient to drive STING polymerization, providing a mechanistic understanding of the requirement for endoplasmic reticulum (ER)-to-Golgi apparatus translocation for STING activation.
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Glicosaminoglicanos/metabolismo , Aparato de Golgi/metabolismo , Proteínas de la Membrana/metabolismo , Nucleótidos Cíclicos/metabolismo , Animales , Células COS , Línea Celular , Línea Celular Tumoral , Chlorocebus aethiops , Cricetinae , Citosol/metabolismo , Retículo Endoplásmico/metabolismo , Células HeLa , Humanos , Ratones , Polimerizacion , Transducción de Señal/fisiología , Transportadores de Sulfato/metabolismo , Vaccinia/metabolismo , Virus Vaccinia/patogenicidadRESUMEN
Post-translational modifications (PTMs) of Cyclin-dependent kinase 5 (CDK5) have emerged as important regulatory mechanisms that modulate cancer development in patients. Though CDK5 is an atypical member of the cyclin-dependent kinase family, its aberrant expression links to cell proliferation, DNA damage response, apoptosis, migration and angiogenesis in cancer. Current studies suggested that, new PTMs on CDK5, including S-nitrosylation, sumoylation, and acetylation, serve as molecular switches to control the kinase activity of CDK5 in the cell. However, a majority of these modifications and their biological significance in cancer remain uncharacterized. In this review, we discussed the role of PTMs on CDK5-mediated signaling cascade, and their possible mechanisms of action in malignant tumors, as well as the challenges and future perspectives in this field. On the basis of the newly identified regulatory signaling pathways of CDK5 related to PTMs, researchers have investigated the cancer therapeutic potential of chemical compounds, small-molecule inhibitors, and competitive peptides by targeting CDK5 and its PTMs. Results of these preclinical studies demonstrated that targeting PTMs of CDK5 yields promising antitumor effects and that clinical translation of these therapeutic strategies is warranted.
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In this study, a novel cationic nanofiber membrane with various functional groups, good structural stability, and high adsorption capacity of Cr(vi) is presented. This nanofiber membrane is prepared by electrospinning a mixed aqueous solution of a cationic polycondensate (CP) and polyvinyl alcohol (PVA). With the aid of PVA, CP can be smoothly electrospun without using any organic solvents, and the cross-linking between CP and PVA improves the stability of membrane in acidic solution. Chemical and morphology characterization reveals that the CP/PVA membrane is composed of interwoven nanofibers that contain numerous cationic groups. Due to its high cationicity and hydrophilicity, the CP/PVA membrane shows great affinity for HCr2O7 - and Cr2O7 2-. Adsorption experiments indicate that the CP/PVA membrane can remove Cr(vi) from simulated wastewater rapidly and efficiently in both batch and continuous mode. Besides, the presence of most coexisting ions will not interfere with the adsorption. Due to the redox reaction between the CP/PVA membrane and adsorbed Cr(vi), the CP/PVA membrane exhibits distinct color change after Cr(vi) adsorption and the discoloration is highly dependent on the adsorption amount. Therefore, in addition to serving as a highly efficient adsorbent, the CP/PVA membrane is also expected to be a convenient and low-cost method for semi-quantitative determination of Cr(vi) in wastewater.
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BACKGROUND: Diosmetin is a bioflavonoid compound naturally abundant in citrus fruits. It is found to perform a variety of activities, while its antitumor property in osteosarcoma, a malignant tumor with unmet clinical treatment, remained unknown. METHODS: Colony formation assay, cell cycle analysis and apoptosis analysis were conducted respectively to observe the effect of diosmetin on cell proliferation and apoptosis in human osteosarcoma cells. Western blot and immunoprecipitation were used to detect the expression of apoptotic molecules and activation of STAT3/c-Myc pathway in Saos-2 and U2SO cells. RESULTS: Diosmetin significantly inhibited cell proliferation, induced cell cycle arrest at G2/M phase and promoted cell apoptosis in both Saos-2 and U2SO cells. Moreover, Diosmetin downregulated the expression of anti-apoptotic protein Bcl-xL while upregulated the levels of pro-apoptotic proteins including cleaved Caspase-3, cleaved-PARP and Bax. Furthermore, diosmetin dose-dependently inhibited STAT3 phosphorylation, reduced the expression of its downstream protein c-Myc and impeded the interaction between STAT3 molecules. CONCLUSIONS: These results suggest that diosmetin exerts anti-osteosarcoma effects by suppressing cell proliferation and inducing apoptosis via inhibiting the activation of STAT3/c-Myc signaling pathway, which provide the possibility for diosmetin to be a chemotherapeutic candidate for osteosarcoma.