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Conversion of sputum from positive to negative is one of the indicators to evaluate the efficacy of anti-tuberculosis treatment (ATT). We investigate the factors associated with delayed sputum conversion after 2 or 5 months of ATT from the perspectives of bacteriology and genomics. A retrospective study of sputum conversion in sputum positive 1782 pulmonary tuberculosis (PTB) was conducted from 2021 to 2022 in Beijing, China. We also designed a case-matched study including 24 pairs of delayed-sputum-conversion patients (DSCPs) and timely-sputum-conversion patients (TSCPs), and collect clinical isolates from DSCPs before and after ATT and initial isolates of TSCPs who successfully achieved sputum conversion to negative after 2 months of ATT. A total of 75 strains were conducted drug sensitivity testing (DST) of 13 anti-TB drugs and whole-genome sequencing (WGS) to analyze the risk factors of delayed conversion and the dynamics changes of drug resistance and genomics of Mycobacterium tuberculosis (MTB) during ATT. We found TSCPs have better treatment outcomes and whose initial isolates show lower levels of drug resistance. Clinical isolates of DSCPs showed dynamically changing of resistance phenotypes and intra-host heterogeneity. Single nucleotide polymorphism (SNP) profiles showed large differences between groups. The study provided insight into the bacteriological and genomic variation of delayed sputum conversion. It would be helpful for early indication of sputum conversion and guidance on ATT.
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Antituberculosos , Genómica , Mycobacterium tuberculosis , Esputo , Tuberculosis Pulmonar , Humanos , Tuberculosis Pulmonar/microbiología , Tuberculosis Pulmonar/tratamiento farmacológico , Tuberculosis Pulmonar/diagnóstico , Esputo/microbiología , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/aislamiento & purificación , Mycobacterium tuberculosis/efectos de los fármacos , Antituberculosos/uso terapéutico , Antituberculosos/farmacología , Masculino , Adulto , Femenino , Estudios Retrospectivos , Persona de Mediana Edad , Genómica/métodos , Polimorfismo de Nucleótido Simple , Pruebas de Sensibilidad Microbiana , Secuenciación Completa del Genoma , Resultado del Tratamiento , Farmacorresistencia Bacteriana/genéticaRESUMEN
Even though robotic-assisted laparoscopic radical prostatectomy (RARP) is superior to open surgery in reducing postoperative complications, 6-20% of patients still experience urinary incontinence (UI) after surgery. Therefore, many researchers have established predictive models for UI occurrence after RARP, but the predictive performance of these models is inconsistent. This study aims to systematically review and critically evaluate the published prediction models of UI risk for patients after RARP. We conducted a comprehensive literature search in the databases of PubMed, Cochrane Library, Web of Science, and Embase. Literature published from inception to March 20, 2024, which reported the development and/or validation of clinical prediction models for the occurrence of UI after RARP. We identified seven studies with eight models that met our inclusion criteria. Most of the studies used logistic regression models to predict the occurrence of UI after RARP. The most common predictors included age, body mass index, and nerve sparing procedure. The model performance ranged from poor to good, with the area under the receiver operating characteristic curves ranging from 0.64 to 0.98 in studies. All the studies have a high risk of bias. Despite their potential for predicting UI after RARP, clinical prediction models are restricted by their limited accuracy and high risk of bias. In the future, the study design should be improved, the potential predictors should be considered from larger and representative samples comprehensively, and high-quality risk prediction models should be established. And externally validating models performance to enhance their clinical accuracy and applicability.
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Laparoscopía , Complicaciones Posoperatorias , Prostatectomía , Procedimientos Quirúrgicos Robotizados , Incontinencia Urinaria , Humanos , Prostatectomía/métodos , Prostatectomía/efectos adversos , Incontinencia Urinaria/etiología , Procedimientos Quirúrgicos Robotizados/efectos adversos , Procedimientos Quirúrgicos Robotizados/métodos , Masculino , Laparoscopía/efectos adversos , Laparoscopía/métodos , Complicaciones Posoperatorias/etiología , Curva ROC , Índice de Masa CorporalRESUMEN
Human exposure to micro- and nano-plastics (MNPs) primarily occurs through respiration and diet in the environment. However, the early effects and warning signs of MNPs exposure on vertebrates are unclear. Here we used intratracheal instillation and intragastric infusion to establish mouse models for MNPs exposure to systematically investigate the toxic mechanisms of MNPs on endocrine organs. Results showed that MNPs induced endocrine disruptions in short-term exposure by both dietary and respiratory pathways. Microplastics (MPs) exposed through dietary route were more toxic to thyroid gland, whereas nanoplastics (NPs) exhibited the highest level of toxicity to parathyroid gland through respiration. The transcriptome and validation of related functional genes revealed that MNPs affected the synthesis of thyroglobulin by interfering with the expressions of PAX8 and CREB. MNPs also impacted the levels of thyroid stimulating hormone, further mediating the secretion of thyroid hormones. Moreover, MNPs modulate the expression of Mafb, thereby exerting regulatory effects on parathyroid hormone (PTH) synthesis and growth development in parathyroid cells. Meanwhile, MNPs interfered with the expression of IP3R in the calcium signaling pathway, indirectly affecting the secretion of PTH. This study reveals the effects and mechanisms of MNPs on thyroid and parathyroid and highlights the significance of early warning of MNPs exposure.
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Microplásticos , Nanopartículas , Factor de Transcripción PAX8 , Glándula Tiroides , Animales , Microplásticos/toxicidad , Glándula Tiroides/efectos de los fármacos , Glándula Tiroides/metabolismo , Nanopartículas/toxicidad , Factor de Transcripción PAX8/genética , Biomarcadores/metabolismo , Glándulas Paratiroides/efectos de los fármacos , Glándulas Paratiroides/metabolismo , Hormona Paratiroidea , Masculino , Ratones , Enfermedades de la Tiroides/inducido químicamente , Enfermedades de las Paratiroides , Tiroglobulina/metabolismo , Tiroglobulina/genéticaRESUMEN
AIM: We reveal the mechanism of action whereby ambient PM2.5 promotes kidney injury. METHODS: Using C57BL/6 mice, the effects of PM2.5 exposure on the acute kidney injury (AKI) were investigated, including renal function changes, expression of inflammatory cytokines, histopathological changes, as well as activation of nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing 3ï¼NLRP3ï¼. The effects of PM2.5 on renal injury after NLRP3 inhibition were explored using NLRP3 inhibitor (MCC950) and NLRP3 knockout mice. The effects of PM2.5 on the inflammatory response of renal macrophages were investigated at the cellular level. RESULTS: PM2.5 exposure could promote kidney injury, NLRP3 activation and inflammatory response in mice. After using MCC950 and NLRP3 knockout mice, the effects of PM2.5 and the kidney injury could be inhibited. The cellular-level results also suggested that MCC950 could inhibit the effects of PM2.5. CONCLUSION: PM2.5 can promote the progression of AKI and aggravate tissue inflammation through NLRP3, which is an important environmental toxicological mechanism of PM2.5.
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Lesión Renal Aguda , Inflamación , Macrófagos , Ratones Endogámicos C57BL , Ratones Noqueados , Proteína con Dominio Pirina 3 de la Familia NLR , Material Particulado , Animales , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Material Particulado/toxicidad , Lesión Renal Aguda/inducido químicamente , Lesión Renal Aguda/patología , Ratones , Macrófagos/efectos de los fármacos , Inflamación/inducido químicamente , Masculino , Sulfonamidas/toxicidad , Sulfonamidas/farmacología , Indenos/toxicidad , Contaminantes Atmosféricos/toxicidad , Furanos/toxicidad , Sulfonas/toxicidadRESUMEN
Targeted next-generation sequencing (tNGS) can be used to perform Mycobacterium tuberculosis (MTB) complex-specific amplification or target capture directly from sputum samples, yielding simultaneous coverage of many genes and DNA regions associated with antimicrobial resistance (AMR). Performance comparisons of tNGS and another molecular testing tool, Xpert MTB/rifampicin (RIF), have been empirical. Here, using a dilution series of a RIF-resistant clinical isolate of MTB, we found that tNGS had a slightly lower limit of bacterial detection (102 CFU/mL) compared with Xpert MTB/RIF (103 CFU/mL) in culture medium. However, the minimum detection limit of the rpoB S450L mutation in this isolate was significantly lower with tNGS (102 CFU/mL) than with Xpert MTB/RIF (106 CFU/mL). Sputum samples collected from 129 suspected pulmonary tuberculosis patients were also prospectively studied with the clinical diagnosis as a reference, revealing that the sensitivity of tNGS (48.6%) was higher than those of culture (46.8%), Xpert MTB/RIF (39.4%), and smear microscopy (34.9%) testing. Notably, AMR analysis of 56 MTB-positive samples as determined by tNGS revealed high mutation frequencies of 96.4%, 35.7%, 26.8%, and 19.6% in the following AMR-associated genes: rrs, rpoB, katG, and pncA, respectively. The findings of this study provide theoretical support for the differential clinical application of tNGS and Xpert MTB/RIF and suggest that tNGS has greater application value in tuberculosis drug resistance monitoring and prevention.IMPORTANCETargeted next-generation sequencing (tNGS) can be used to perform Mycobacterium tuberculosis (MTB) complex-specific amplification or target capture directly from sputum samples, yielding simultaneous coverage of genes and DNA regions associated with antimicrobial resistance (AMR). Performance comparisons of tNGS and Xpert MTB/rifampicin (RIF) have been empirical. The Xpert MTB/RIF assay is a commercial system that uses the nucleic acid amplification detection method for rapid (2 hours) diagnosis of tuberculosis (TB). The cost of the tNGS and Xpert MTB/RIF assays in this study was similar, at USD 98 and USD 70-104 per sample, respectively, but the time required for tNGS (3 days) was much longer than that required for the Xpert MTB/RIF assay. However, tNGS yielded more accurate results and a larger number of AMR-associated gene mutations, which compensated for the extra time and highlighted the greater application value of tNGS in TB drug resistance monitoring and prevention.
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Secuenciación de Nucleótidos de Alto Rendimiento , Mycobacterium tuberculosis , Rifampin , Esputo , Tuberculosis Pulmonar , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/aislamiento & purificación , Humanos , Esputo/microbiología , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Rifampin/farmacología , Tuberculosis Pulmonar/diagnóstico , Tuberculosis Pulmonar/microbiología , Sensibilidad y Especificidad , Tuberculosis Resistente a Múltiples Medicamentos/diagnóstico , Tuberculosis Resistente a Múltiples Medicamentos/microbiología , Proteínas Bacterianas/genética , Mutación , Farmacorresistencia Bacteriana/genética , Técnicas de Diagnóstico Molecular/métodos , Pruebas de Sensibilidad Microbiana , Femenino , ARN Polimerasas Dirigidas por ADN/genética , Masculino , Adulto , ADN Bacteriano/genéticaRESUMEN
Toxic organic pollutants in wastewater have seriously damaged human health and ecosystems. Photocatalytic degradation is a potential and efficient tactic for wastewater treatment. Among the entire carbon family, biochar has been developed for the adsorption of pollutants due to its large specific surface area, porous skeleton structure, and abundant surface functional groups. Hence, combining adsorption and photocatalytic decomposition, TiO2-biochar photocatalysts have received considerable attention and have been extensively studied. Owing to biochar's adsorption, more active sites and strong interactions between contaminants and photocatalysts can be achieved. The synergistic effect of biochar and TiO2 nanomaterials substantially improves the photocatalytic capacity for pollutant degradation. TiO2-biochar composites have numerous attractive properties and advantages, culminating in infinite applications. This review discusses the characteristics and preparation techniques of biochar, presents in situ and ex situ synthesis approaches of TiO2-biochar nanocomposites, explains the benefits of TiO2-biochar-based compounds for photocatalytic degradation, and emphasizes the strategies for enhancing the photocatalytic efficiency of TiO2-biochar-based photocatalysts. Finally, the main difficulties and future advancements of TiO2-biochar-based photocatalysis are highlighted. The review gives an exhaustive overview of recent progress in TiO2-biochar-based photocatalysts for organic contaminants removal and is expected to encourage the development of robust TiO2-biochar-based photocatalysts for sewage remediation and other environmentally friendly uses.
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OBJECTIVES: We elucidated the factors, evolution, and compensation of antimicrobial resistance (AMR) in Mycobacterium tuberculosis (MTB) isolates under dual pressure from the intra-host environment and anti-tuberculosis (anti-TB) drugs. METHODS: This retrospective case-control study included 337 patients with pulmonary tuberculosis from 15 clinics in Tianjin, China, with phenotypic drug susceptibility testing results available for at least two time points between January 1, 2009 and December 31, 2016. Patients in the case group exhibited acquired AMR to isoniazid (INH) or rifampicin (RIF), while those in the control group lacked acquired AMR. The whole-genome sequencing (WGS) was conducted on 149 serial longitudinal MTB isolates from 46 patients who acquired or reversed phenotypic INH/RIF-resistance during treatment. The genetic basis, associated factors, and intra-host evolution of acquired phenotypic INH/RIF-resistance were elucidated using a combined analysis. RESULTS: Anti-TB interruption duration of ≥30 days showed association with acquired phenotypic INH/RIF resistance (aOR = 2·2, 95% CI, 1·0-5·1) and new rpoB mutations (p = 0·024). The MTB evolution was 1·2 (95% CI, 1·02-1·38) single nucleotide polymorphisms per genome per year under dual pressure from the intra-host environment and anti-TB drugs. AMR-associated mutations occurred before phenotypic AMR appearance in cases with acquired phenotypic INH (10 of 16) and RIF (9 of 22) resistances. DISCUSSION: Compensatory evolution may promote the fixation of INH/RIF-resistance mutations and affect phenotypic AMR. The TB treatment should be adjusted based on gene sequencing results, especially in persistent culture positivity during treatment, which highlights the clinical importance of WGS in identifying reinfection and AMR acquisition before phenotypic drug susceptibility testing.
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Antituberculosos , Isoniazida , Mycobacterium tuberculosis , Rifampin , Tuberculosis Pulmonar , Secuenciación Completa del Genoma , Humanos , Mycobacterium tuberculosis/efectos de los fármacos , Mycobacterium tuberculosis/genética , Antituberculosos/farmacología , Antituberculosos/uso terapéutico , Estudios Retrospectivos , Masculino , Femenino , Persona de Mediana Edad , Adulto , Estudios de Casos y Controles , Rifampin/farmacología , Rifampin/uso terapéutico , Tuberculosis Pulmonar/tratamiento farmacológico , Tuberculosis Pulmonar/microbiología , Isoniazida/farmacología , Isoniazida/uso terapéutico , China , Pruebas de Sensibilidad Microbiana , Tuberculosis Resistente a Múltiples Medicamentos/tratamiento farmacológico , Tuberculosis Resistente a Múltiples Medicamentos/microbiología , Fenotipo , Mutación , Farmacorresistencia Bacteriana/genética , Anciano , Evolución Molecular , Proteínas Bacterianas/genética , Farmacorresistencia Bacteriana Múltiple/genéticaRESUMEN
Introduction: Mycobacterium tuberculosis (MTB) has a type III-A clustered regularly interspaced short palindromic repeat/CRISPR-associated protein (CRISPR/Cas) system consisting of a Csm1-5 and CRISPR RNA (crRNA) complex involved in the defense against invading nucleic acids. However, CRISPR/Cas system in the MTB still is clearly unknown and needs to be further explored. Methods: In our work, two non-Cas system proteins EspB and HtpG protein were found and identified by LC-MS/MS. The effect of EspB and HtpG on Type III-A CRISPR/Cas System of M. tuberculosis was examined by using Plasmid interference assay and Co-immunoprecipitation analyses. We explored that EspB could interact with the crRNA RNP complex, but HtpG could inhibit the accumulation of the MTB Csm proteins and defense the mechanism of CRISPR/Cas system. Results: The proteins ESAT-6 secretion system-1(Esx-1) secreted protein B (EspB) and high-temperature protein G (HtpG), which were not previously associated with CRISPR/Cas systems, are involved in mycobacterial CRISPR/Cas systems with distinct functions. Conclusion: EspB is a novel crRNA-binding protein that interacts directly with the MTB crRNP complex. Meanwhile, HtpG influences the accumulation of MTB Csm proteins and EspB and interferes with the defense mechanism of the crRNP complex against foreign DNA in vivo. Thereby, our study not only leads to developing more precise clinical diagnostic tool to quickly detect for MTB infection, but also knows these proteins merits for TB biomarkers/vaccine candidates.
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Background and Objective: This study was performed to investigate the association of peripheral T lymphocyte subsets with disseminated infection (DI) by Mycobacterium tuberculosis (MTB) in HIV-negative patients. Methods and Materials: The study included 587 HIV-negative tuberculosis (TB) patients. Results: In TB patients with DI, the proportion of CD4+ T cells decreased, the proportion of CD8+ T cells increased, and the ratio of CD4+/CD8+ T cells decreased. According to univariate analysis, smoking, alcohol consumption, rifampicin-resistance, retreatment, and high sputum bacterial load were linked to lower likelihood of developing MTB dissemination. Multivariate analysis indicated that after adjustment for alcohol use, smoking, retreatment, smear, culture, rifampicin-resistance, and CD4+/CD8+, the proportion of CD8+ T cells (but not CD4+ T cells) was independently and positively associated with the prevalence of DI in HIV-negative pulmonary TB (PTB) patients. Conclusions: Examining T lymphocyte subsets is of great value for evaluating the immune function of HIV-negative TB patients, and an increase in the CD8+ T cell proportion may be a critical clue regarding the cause of DI in such patients.
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Infecciones por VIH , Mycobacterium tuberculosis , Tuberculosis Ganglionar , Humanos , Rifampin , Subgrupos de Linfocitos T , Infecciones por VIH/complicacionesRESUMEN
The need for novel antibiotics has become imperative with the increasing prevalence of antibiotic resistance in Gram-negative bacteria in clinics. Acting as a permeability barrier, lipopolysaccharide (LPS) protects Gram-negative bacteria against drugs. LPS is synthesized in cells and transported to the outer membrane (OM) via seven lipopolysaccharide transport (Lpt) proteins (LptA-LptG). Of these seven Lpt proteins, LptC interacts with LptA to transfer LPS from the inner membrane (IM) to the OM, and assembly is aided by LptD/LptE. This interaction among the Lpt proteins is important for the biosynthesis of LPS; therefore, the Lpt proteins, which are significant in the assembly process of LPS, can be a potential target for new antibiotics. In this study, a yeast two-hybrid (Y2H) system was used to screen compounds that could block LPS transport by inhibiting LptA/LptC interaction, which finally disrupts the biosynthesis of the OM. We selected the compound IMB-0042 for this study. Our results suggest that IMB-0042 disrupts LptA/LptC interaction by binding to both LptA and LptC. Escherichia coli cells, when treated with IMB-0042, showed filament morphology, impaired OM integrity, and an accumulation of LPS in the periplasm. IMB-0042 inhibited the growth of Gram-negative bacteria and showed synergistic sensitization to other antibiotics, with low cytotoxicity. Thus, we successfully identified a potential antibacterial agent by using a Y2H system, which blocks the transport of LPS by targeting LptA/LptC interaction in Escherichia coli.
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Karanjin is a bioactive furanoflavonoid with various pharmacological activities including anticancer activities. However, the effect and the related mechanism of karanjin in breast cancer (BC) have not been revealed. The potential targets of karanjin and BC were predicted using SwissTargetPrediction and GeneCards databases, respectively. The overlapping targets between karanjin and BC were identified using the Venn diagram. DAVID database was used for the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathway analysis. Cell viability, proliferation, and apoptosis were examined by MTT (3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-tetrazolium bromide), EdU (5-ethynyl-2'-deoxyuridine) incorporation, and TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP digoxigenin nick-end labeling) assays, respectively. The protein levels were measured by western blot analysis. We screened out 28 overlapping targets between karanjin and BC. KEGG analysis showed that the targets of karanjin in BC were associated with the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway. Karanjin inhibited cell viability and impeded the proliferative ability of BC cells. Moreover, karanjin treatment induced apoptosis in BC cells. Additionally, karanjin treatment blocked the PI3K/Akt signaling pathway and activation of the PI3K/Akt pathway reversed the antitumor effect of karanjin on BC cells. In conclusion, karanjin exerted antitumor activity in BC cells by regulating the PI3K/Akt signaling pathway.
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Neoplasias de la Mama , Proteínas Proto-Oncogénicas c-akt , Humanos , Femenino , Proteínas Proto-Oncogénicas c-akt/metabolismo , Fosfatidilinositol 3-Quinasa/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Transducción de Señal , Apoptosis , Proliferación CelularRESUMEN
ATP-binding cassette (ABC) subfamily A member 8 (ABCA8) has been reported to play a vital role in cancer development. Our study aimed to explore the role and the molecular mechanism of ABCA8 in breast cancer (BC) progression. GSE65194, GSE15852, and GSE45827 datasets were used to identify differentially expressed genes (DEGs) in BC. The diagnosis and prognosis value were determined using ROC curve analysis and Kaplan-Meier plotter, respectively. The relationship between ABCA8 expression and clinicopathological features in BC was analyzed by TCGA. Co-expressed genes of ABCA8 in BC were screened out through GEPIA and subjected to KEGG pathway enrichment analysis. Cell proliferation was evaluated by CCK-8 and EdU incorporation assays. Proliferating cell nuclear antigen (PCNA) expression and the changes of the AMP activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway were measured by western blot analysis. Totally 4 overlapping DEGs were identified and all reduced in BC samples. ABCA8 with high diagnostic and prognostic values was selected for further exploration. Low ABCA8 expression was correlated with clinicopathological features in BC patients. ABCA8 overexpression inhibited BC cell proliferation. The top 20 co-expressed genes of ABCA8 were identified by GEPIA and significantly enriched in AMPK signaling pathway. Inhibition of AMPK/mTOR pathway reversed the suppressive effect of ABCA8 on BC cell growth. These results suggested that ABCA8 overexpression repressed BC cell proliferation through regulating the AMPK/mTOR signaling pathway.
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Proteínas Quinasas Activadas por AMP , Neoplasias de la Mama , Proteínas Quinasas Activadas por AMP/genética , Proteínas Quinasas Activadas por AMP/metabolismo , Transportadoras de Casetes de Unión a ATP/genética , Transportadoras de Casetes de Unión a ATP/metabolismo , Neoplasias de la Mama/metabolismo , Proliferación Celular/genética , Femenino , Humanos , Transducción de Señal , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
In Gram-negative bacteria, lipopolysaccharide transport (Lpt) protein LptA and LptC form a complex to transport LPS from the inner membrane (IM) to the outer membrane (OM). Blocking the interaction between LptA and LptC will lead to the defect of OM and cell death. Therefore, Lpt protein interaction could be used as a target to screen new drugs for killing Gram-negative bacteria. Here we used biolayer interferometry (BLI) assay to detect the interaction between LptA and LptC, with the aim to develop a method for screening the LptA/LptC interaction blockers in vitro. Firstly, LptC and LptA with or without signal peptide (LptAfull or LptAno signal) were expressed in E. coli BL21(DE3). The purified proteins were then labeled with biotin and the super streptavidin (SSA) biosensor was blocked with diluent. The biotin labeled protein sample was mixed with the sensor, and then the binding of the protein with a series of diluted non biotinylated protein was detected. At the same time, non-biotinylated protein was used as a control. The binding of biotinylated protein to a small molecule IMB-881 and the blocking of interaction were also detected by the same method. In the blank control, the biosensor without biotinylated protein was used to detect the serially diluted samples. The signal response constant was calculated by using steady analysis. The results showed that biotinylated LptC had a good binding activity with LptAfull and LptAno signal with KD value 2.9eâ»7±7.9eâ»8 and 6.0eâ»7±2.8eâ»8, respectively; biotinylated LptAno signal had a good binding activity with LptC, with a KD value of 9.6eâ»7±7.2eâ»8. All binding curves showed obvious fast binding and fast dissociation morphology. The small molecule compound IMB-881 can bind to LptA to block the interaction between LptA and LptC, but has no binding activity with LptC. In summary, we developed a method for detecting the LptA/LptC interaction based on the BLI technology, and confirmed that this method can be used to evaluate the blocking activity of small molecule blockers, providing a new approach for the screening of LptA/LptC interaction blockers.
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Proteínas de Escherichia coli , Escherichia coli , Proteínas Portadoras , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Interferometría , Proteínas de la Membrana/metabolismoRESUMEN
The development of novel treatments for lymphedema is hindered by the poorly understood pathophysiology of the disease. To improve the therapeutic success of treating the disease, the present study aimed to investigate the effects and mechanism of long noncoding RNA myocardial infarctionassociated transcript (MIAT) in terms of the differentiation of adiposederived mesenchymal stem cells (ADMSCs) into lymphatic endothelial cells (LECs). The expression levels of (MIAT), microRNA (miR)495 and Prosperorelated homeobox 1 (Prox1) were measured by reverse transcriptionquantitative PCR. The protein expression levels of Prox1, lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1), vascular endothelial growth factor receptor3 (VEGFR3) and podoplanin (PDPL) were detected by western blotting and immunofluorescence. A dualluciferase reporter assay was also used to detect the interaction between MIAT, miR495 and Prox1. In addition, migration and tubeformation capabilities were measured by Transwell assay and tubeformation assay, respectively. The results obtained demonstrated that VEGFC156S (recombinant VEGFC in which Cys156 was replaced by Ser residue) treatment could efficiently induce the differentiation of ADMSCs into LECs. MIAT expression was upregulated and miR495 was downregulated during differentiation. Mechanistically, MIAT upregulated Prox1 expression possibly by acting as a molecular sponge for miR495. Functional analyses indicated that the expression levels of Prox1, LYVE1, VEGFR3 and PDPL, and the migration and tubeformation capabilities of ADMSCs induced by VEGFC156S, were significantly inhibited by silencing MIAT and overexpressing miR495. Moreover, miR495 inhibition and Prox1 overexpression reversed the effects of MIAT downregulation and miR495 upregulation, respectively, on the differentiation of ADMSCs into LECs. Taken together, these results suggested that MIAT may be involved in the differentiation of ADMSCs into LECs, and that the MIAT/miR495/Prox1 axis may be a novel regulatory mechanism and therapeutic target for the treatment of lymphedema.
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Proteínas de Homeodominio/genética , Células Madre Mesenquimatosas/metabolismo , MicroARNs/genética , Infarto del Miocardio/genética , ARN Largo no Codificante/genética , Proteínas Supresoras de Tumor/genética , Animales , Diferenciación Celular/genética , Movimiento Celular/genética , Células Endoteliales/citología , Células Endoteliales/metabolismo , Humanos , Células Madre Mesenquimatosas/citología , Infarto del Miocardio/patología , Factor A de Crecimiento Endotelial Vascular/genética , Proteínas de Transporte Vesicular/genéticaRESUMEN
OBJECTIVE: To observe the clinical effect of Yihechun Capsules (YHC) on oligozoospermia and asthenospermia. METHODS: A total of 181 male patients with infertility were randomly divided into a YHC+Levocarnitine (LC) group (n = 93, including 42 cases of oligozoospermia, 20 cases of asthenospermia and 31 cases of oligoasthenospermia) and an LC control group (n = 88, including 39 cases of oligozoospermia, 22 cases of asthenospermia and 27 cases of oligoasthenospermia), the former treated with YHC (ï¼»0.3 g per capsuleï¼½, once 4 capsules, bid, 30 minutes after meal) combined with LC oral liquid (2ï¼3 g/d, tid, at mealtime) and the latter with LC oral liquid only (2ï¼3 g/d, tid, at mealtime). After 3 months of treatment, comparisons were made between the two groups of patients in sperm concentration, the percentages of grade a and grade a+b sperm, and the rate of pregnancy. RESULTS: Of the 181 patients, 5 in the YHC+LC group and 2 in the LC control group failed to complete the course of treatment. There were no statistically significant differences between the two groups of patients in the baseline sperm concentration and the percentages of grade a and grade a+b sperm (P > 0.05), wich were all markedly increased in both the YHC+LC and the LC control groups (P < 0.05) after 3 months of treatment. And the patients of the YHC+LC group, compared with the controls, showed even more significant increases, as the oligozoospermia patients in sperm concentration (ï¼»21.07 ± 6.98ï¼½ vs ï¼»16.56 ± 1.82ï¼½ ×106/ml, P < 0.05) and the percentages of grade a sperm (ï¼»27.53 ± 3.34ï¼½% vs ï¼»26.88 ± 1.35ï¼½%, P < 0.05) and grade a+b sperm (ï¼»53.32 ± 3.16ï¼½% vs ï¼»52.63 ± 2.48ï¼½%, P < 0.05), the asthenospermia patients in sperm concentration (ï¼»26.36 ± 3.37ï¼½ vs ï¼»24.42 ± 2.21ï¼½ ×106/ml, P < 0.05) and the percentages of grade a sperm (ï¼»25.28 ± 4.64ï¼½% vs ï¼»21.32 ± 3.28ï¼½%, P < 0.05) and grade a+b sperm (ï¼»49.19 ± 2.87ï¼½% vs ï¼»45.64 ± 1.78ï¼½%, P < 0.05), and the oligoasthenospermia patients in sperm concentration (ï¼»19.38 ± 3.39ï¼½ vs ï¼»18.75 ± 1.35ï¼½ ×106/ml, P < 0.05) and the percentages of grade a sperm (ï¼»22.65 ± 4.81ï¼½% vs ï¼»21.31 ± 2.42ï¼½%, P < 0.05) and grade a+b sperm (ï¼»48.74 ± 5.61ï¼½% vs ï¼»44.36 ± 1.32ï¼½%, P < 0.05). The pregnancy rate was dramatically higher in the YHC+LC than in the LC control group (36.4% ï¼»32/88ï¼½ vs 15.1% ï¼»13/86ï¼½, P < 0.01). CONCLUSIONS: Yihechun Capsules combined with Levocarnitine oral liquid is evidently effective for the treatment of oligozoospermia and asthenospermia.
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Astenozoospermia/tratamiento farmacológico , Carnitina/uso terapéutico , Medicamentos Herbarios Chinos/uso terapéutico , Oligospermia/tratamiento farmacológico , Femenino , Humanos , Masculino , Embarazo , Índice de Embarazo , Recuento de Espermatozoides , Motilidad Espermática , EspermatozoidesRESUMEN
Hepatocellular carcinoma (HCC) is a major leading cause of cancer-related death worldwide. Alpha fetoprotein (AFP) is reactivated in a majority of hepatocellular carcinoma (HCC) and associated with poor patient outcomes. Although increasing evidence has shown that AFP can regulate HCC cell growth, the precise functions of AFP in hepatocarcinogenesis and the associated underlying mechanism remain incompletely understood. In this study, we demostrated that depleting AFP significantly suppressed diethylnitrosamine (DEN)-induced liver tumor progression in an AFP gene-deficient mouse model. Similarly, knocking down AFP expression inhibited human HCC cell proliferation and tumor growth by inducing apoptosis. AFP expression level was inversely associated with the apoptotic rate in mouse and human HCC specimens. Investigation of potential cross-talk between AFP and apoptotic signaling revealed that AFP exerted its growth-promoting effect by suppressing the Fas/FADD-mediated extrinsic apoptotic pathway. Mechanistically, AFP bound to the RNA-binding protein HuR, increasing the accumulation of HuR in the cytoplasm and subsequent inhibition of Fas mRNA translation. In addition, we found that inhibiting AFP enhanced the cytotoxicity of therapeutics to AFP-positive HCC cells by activating HuR-mediated Fas/FADD apoptotic signaling. Conclusion: Our study defined the pro-oncogenic role of AFP in HCC progression and uncovered a novel antiapoptotic mechanism connecting AFP to HuR-mediated Fas translation. Our findings suggest that AFP is involved in the pathogenesis and chemosensitivity of HCC and that blockade of AFP may be a promising strategy to treat advanced HCC.
Asunto(s)
Apoptosis/genética , Carcinoma Hepatocelular/metabolismo , Proteína de Dominio de Muerte Asociada a Fas/metabolismo , alfa-Fetoproteínas/genética , Animales , Carcinogénesis/genética , Carcinoma Hepatocelular/patología , Línea Celular Tumoral , Proteína de Dominio de Muerte Asociada a Fas/genética , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patología , Ratones , alfa-Fetoproteínas/metabolismoRESUMEN
BACKGROUND: Most studies have showed that maternal depression is associated with pregnancy complications. However, there were limited evidences in Chinese population. We examined the associations of antenatal depression symptoms with pregnancy outcomes, especially for low birth weight. METHODS: A total of 1377 singleton pregnant women were recruited from Nanshan Maternity & Child Healthcare Hospital of Shenzhen in this prospective cohort study. Depression symptoms were assessed by the Edinburgh postnatal depression scale (EPDS) questionnaire in the second trimester of gestation; cut-points for the indication of antenatal depression were â§12 scores in this study. Socio-demographic data, life-style and pregnancy outcomes were collected through Shenzhen Maternity & Child Healthcare database. The risks of adverse outcomes in pregnant women with antenatal depression were determined by multivariate logistic regression and represented as odds ratio(OR) and 95% confidence interval (CI). RESULTS: Of the 1377 subjects, the prevalence of antenatal depression was 19.1%. The EPDS scores were 13.8 ± 2.0 and 6.5 ± 2.9 (P < 0.001) in subjects with and without antenatal depression, respectively. After adjustment for maternal age, education, parity, pre-pregnancy body mass index (BMI), residential area, fetal gender, an EPDS score ≥ 12 (versus. < 12) was associated with an increased risk for low birth weight (odds ratio: 2.05, 95% CI: 1.12-4.64), but not for preterm birth, large for gestational age, small for gestational age or macrosomia. CONCLUSION: Pregnant women presenting antenatal depressive symptoms are at elevated risk of low birth weight. Mental health problems of pregnancy should be addressed for the prevention of low birth weight.
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Depresión/complicaciones , Depresión/epidemiología , Complicaciones del Embarazo/epidemiología , Complicaciones del Embarazo/psicología , Adulto , China , Depresión/diagnóstico , Femenino , Humanos , Recién Nacido de Bajo Peso , Recién Nacido , Modelos Logísticos , Masculino , Oportunidad Relativa , Paridad , Embarazo , Complicaciones del Embarazo/diagnóstico , Resultado del Embarazo , Estudios Prospectivos , Factores de Riesgo , Adulto JovenRESUMEN
OBJECTIVE: To study the effect of Compound Amino Acid Capsules (CAAC) combined with clomiphene in the treatment of severe oligospermia. METHODS: A total of 104 patients with severe oligospermia admitted to our Center of Reproductive Medicine from January to September 2018 were randomly assigned to a trial (n = 60) and a control group (n = 44), the former treated by oral administration of CAAC combined with clomiphene and the latter with clomiphene only, both for 12 weeks. Comparisons were made between the two groups of patients in the sperm concentration, and the percentages of progressively motile sperm (PMS) and total motile sperm (TMS) before and after 4, 8 and 12 weeks of medication as well as the pregnancy rate during the treatment. RESULTS: Compared with the baseline, the trial group showed significant elevation at 4, 8 and 12 weeks in sperm concentration (ï¼»3.13 ± 1.29ï¼½ vs ï¼»12.06 ± 2.24ï¼½, ï¼»22.10 ± 2.65ï¼½ and ï¼»28.13 ± 3.59ï¼½ ×106/ml, P < 0.01), PMS (ï¼»14.03 ± 2.49ï¼½% vs ï¼»21.05 ± 3.14ï¼½%, ï¼»29.08 ± 4.70ï¼½% and ï¼»35.08 ± 3.70ï¼½%, P < 0.01) and TMS (ï¼»20.10 ± 4.05ï¼½% vs ï¼»27.10 + 4.87ï¼½%, ï¼»36.09 ± 5.64ï¼½% and ï¼»45.04 ± 6.69ï¼½%, P < 0.01), and so did the control group in sperm concentration (ï¼»3.27 ± 1.46ï¼½ vs ï¼»10.21 ± 2.35ï¼½, ï¼»19.89 ± 2.74ï¼½ and ï¼»25.23 ± 3.69ï¼½ ×106/ml, P < 0.01), PMS (ï¼»13.32 ± 3.12ï¼½% vs ï¼»17.02 ± 3.26ï¼½%, ï¼»22.13 ± 3.70ï¼½% and ï¼»27.18 ± 2.54ï¼½%, P < 0.01) and TMS (ï¼»21.30 ± 4.87ï¼½% vs ï¼»24.22 ± 5.07ï¼½%, ï¼»30.03 ± 5.33ï¼½% and ï¼»35.05 ± 5.69ï¼½%, P < 0.01), even more significant in the trial than in the control group at the three time points after medication (P < 0.01). The pregnancy rate was markedly higher in the former than in the latter group at 4 (1.72% vs 0.53%, P < 0.01), 8 (4.21% vs 2.87%, P < 0.01) and 12 weeks (8.32% vs 6.32%, P < 0.01). No adverse reactions were observed in neither of the two groups during the treatment. CONCLUSIONS: CAAC combined with clomiphene can significantly improve the semen parameters of the patients with severe oligospermia, with no obvious adverse events.
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Aminoácidos/administración & dosificación , Clomifeno/administración & dosificación , Oligospermia/terapia , Cápsulas , Femenino , Humanos , Masculino , Embarazo , Índice de Embarazo , Recuento de EspermatozoidesRESUMEN
BACKGROUND: Polycystic ovary syndrome (PCOS) is a heterogeneous endocrine disorder in women of reproductive age and is commonly complicated by adverse endometrial outcomes. Long non-coding RNAs (lncRNAs) are a class of non-protein-coding transcripts that are more than 200 nucleotides in length. Accumulating evidence indicates that lncRNAs are involved in the development of various human diseases. Among these lncRNAs, lncRNA CD36-005 (CD36-005) is indicated to be associated with the pathogenesis of PCOS. However, the mechanisms of action of CD36-005 have not yet been elucidated. METHODS: This study determined the CD36-005 expression level in the uteri of PCOS rat model and its effect on the proliferation activity of rat primary endometrial stromal cells. RNA sequencing (RNA-seq) and bioinformatics analysis were performed to detect the mRNA expression profiles and the biological pathways in which these differentially expressed mRNAs involved, after CD36-005 overexpression in the primary endometrial stromal cells. The differential expression of Hmgn5, Nr5a2, Dll4, Entpd1, Fam50a, and Brms1 were further validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). RESULTS: CD36-005 is highly expressed in the uteri of PCOS rat model and promotes the proliferation of rat primary endometrial stromal cells. A total of fifty-five mRNAs differentially expressed were identified in CD36-005 overexpressed stromal cells. Further analyses identified that these differentially expressed mRNAs participate in many biological processes and are associated with various human diseases. The results of qRT-PCR validation were consistent with the RNA-seq data. CONCLUSIONS: These data provide a list of potential target mRNA genes of CD36-005 in endometrial stromal cells and laid a foundation for further studies on the molecular function and mechanism of CD36-005 in the endometrium.