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The clinical heterogeneity of early-stage endometrial cancer (EC) is worthy of further study to identify high-quality prognostic markers and their potential role in aggressive tumor behavior. Mutation of TP53 was considered as an important primary triage in modified molecular typing for EC, it still cannot precisely predict the prognosis of EC. After proteomic analysis of cancer and para-cancerous tissues from 24 early-stage endometrioid EC patients with different survival outcomes, 13 differentially expressed proteins were screen out while 2 proteins enriched in p53 signaling pathway were further identified by single-cell transcriptome (scRNA-seq). Interestingly, tumor necrosis factor type-1 receptor-associated protein (TRAP1) and calmodulin-regulated spectrin-associated protein family member 3 (CAMSAP3) were found to be significantly downregulated in the specific cell cluster. Expectedly, the signature genes of TRAP1low/CAMSAP3low cluster included classical oncogenes. Moreover, close cellular interactions were observed between myeloid cells and the TRAP1low/CAMSAP3low cluster after systematically elucidating their relationship with tumor microenvironment (TME). The expression of TRAP1 and CAMSAP3 was verified by immunohistochemistry. Thus, a novel prediction model combining TRAP1, CAMSAP3 and TP53 was construct by multi-omics. Compared with the area under the curve, it demonstrated a significantly improvemrnt in the diagnostic efficacy in EC patients from TCGA bank. In conclusion, this work improved the current knowledge regarding the prognosis of early-stage EC through proteomics and scRNA-seq. These findings may lead to improvements in precise risk stratification of early-stage EC patients.
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Biomarcadores de Tumor , Neoplasias Endometriales , Regulación Neoplásica de la Expresión Génica , Estadificación de Neoplasias , Proteómica , Humanos , Femenino , Neoplasias Endometriales/genética , Neoplasias Endometriales/patología , Neoplasias Endometriales/metabolismo , Neoplasias Endometriales/mortalidad , Pronóstico , Biomarcadores de Tumor/genética , Proteómica/métodos , Microambiente Tumoral/genética , Perfilación de la Expresión Génica , Persona de Mediana Edad , Transcriptoma , Multiómica , Proteínas HSP90 de Choque TérmicoRESUMEN
PURPOSE: Few studies have focused on the impact of human papillomavirus (HPV) positivity in male partners on female HPV infection and cervical lesions. The purpose of this study was to evaluate the impact of the HPV infection status of husbands on wives' cervical HPV infection and lesions. METHODS: We surveyed 251 monogamous couples who attended the outpatient department of Fujian Maternity and Child Health Hospital from 2013 to 2021. HPV type analysis was performed on exfoliated cells of the females' cervix and males' urethra by the PCR-reverse dot blot method. We analyzed the prevalence and consistency of HPV types in 251 couples. Subsequently, the risk of HPV infection in females with HPV-positive male partners was analyzed. SPSS version 26 (IBM, Chicago, USA) was used for statistical analysis. RESULTS: In 251 couples, the most commonly detected high-risk HPV (HR-HPV) genotypes were 52, 51, 16, and 58 for males and 16, 52, 18, and 58 for females. Wives with HPV-positive husbands had higher infection rates for most HR-HPV genotypes. HR-HPV positivity in husbands was a risk factor for the development of cervical lesions in wives (OR = 2.250, P = 0.014). Both single-type (OR = 2.085, P = 0.040) and multiple-type (OR = 2.751, P = 0.036) infection in husbands will contributed to an increased risk of non-HR-HPV infection and cervical lesions in wives. CONCLUSION: Husbands' HPV positivity increases the burden of non-HR-HPV infection and increases the risk of cervical lesions developing in wives. It is hoped to provide a reference value for cervical cancer prevention in females and HPV vaccination in males.
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Infecciones por Papillomavirus , Neoplasias del Cuello Uterino , Embarazo , Niño , Humanos , Masculino , Femenino , Heterosexualidad , Infecciones por Papillomavirus/epidemiología , Papillomaviridae/genética , Cuello del Útero , Genotipo , Prevalencia , Neoplasias del Cuello Uterino/epidemiologíaRESUMEN
Cry2Ab is one of the important alternative Bt proteins that can be used to manage insect pests resistant to Cry1A toxins and to expand the insecticidal spectrum of pyramided Bt crops. Previous studies have showed that vacuolar H+-ATPase subunits A and B (V-ATPase A and B) may be involved in Bt insecticidal activities. The present study investigated the role of V-ATPases subunit E in the toxicity of Cry2Ab in Helicoverpa amigera. RT-PCR analysis revealed that oral exposure of H. amigera larvae to Cry2Ab led to a significant reduction in the expression of H. armigera V-ATPase E (HaV-ATPase E). Ligand blot, homologous and heterologous competition experiments confirmed that HaV-ATPases E physically and specifically bound to activated Cry2Ab toxin. Heterologous expressing of HaV-ATPase E in Sf9 cells made the cell line more susceptible to Cry2Ab, whereas knockdown of the endogenous V-ATPase E in H. zea midgut cells decreased Cry2Ab's cytotoxicity against this cell line. Further in vivo bioassay showed that H. armigera larvae fed a diet overlaid with both Cry2Ab and E. coli-expressed HaV-ATPase E protein suffered significantly higher mortality than those fed Cry2Ab alone. These results support that V-ATPases E is a putative receptor of Cry2Ab and can be used to improve Cry2Ab toxicity and manage Cry2Ab resistance at least in H. armigera.
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Bacillus thuringiensis , Insecticidas , Mariposas Nocturnas , Animales , Helicoverpa armigera , Endotoxinas/toxicidad , Endotoxinas/metabolismo , Adenosina Trifosfatasas/genética , Adenosina Trifosfatasas/metabolismo , Escherichia coli , Toxinas de Bacillus thuringiensis/metabolismo , Mariposas Nocturnas/genética , Mariposas Nocturnas/metabolismo , Larva/metabolismo , Insecticidas/toxicidad , Insecticidas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/toxicidad , Proteínas Hemolisinas/metabolismo , Bacillus thuringiensis/metabolismo , Resistencia a los InsecticidasRESUMEN
Low-molecular-weight (LMW, <1000 Da) dissolved organic matter (DOM) plays a significant role in metal/organic pollutant complexation, as well as photochemical/microbiological processes in freshwater ecosystems. The micro size and high reactivity of LMW-DOM hinder its precise characterization. In this study, Suwannee River fulvic acid (SRFA), a commonly used reference material for aquatic DOM, was applied to examine the optical features and molecular composition of LMW-DOM by combining membrane separation, ultraviolet-visible absorption and Orbitrap mass spectrometry (MS) characterization. The 100-500 Da molecular weight cut-off (MWCO) membrane had a better performance in regard to separating the tested LMW-DOM relative to the 500-1000 Da MWCO membrane. The ultraviolet-visible absorbance decreased dramatically for the retentates, whereas it increased for the dialysates. Specifically, carbohydrates, lipids and peptides exhibited high selectivity to the 100-500 Da MWCO membrane in early dialysis. Lignins, tannins and condensed aromatic molecules displayed high permeability to the 500-1000 Da MWCO membrane in late dialysis. Overall, the retentates were dominated by aromatic rings and phenolic hydroxyls with high O/Cwa (weighted average of O/C) and low H/Cwa. Conversely, such dialysates had numerous aliphatic chains with high H/Cwa and low O/Cwa compared to SRFA. In particular, LMW-DOM below 200 Da was identified by Orbitrap MS. This work provides an operational program for identifying LMW-DOM based on the SRFA standard and MS analysis.
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This study focused on constructing iron(III)-tetraamidomacrocyclic ligand (FeIII-TAML)-based magnetic nanostructures via a surfactant-assisted self-assembly (SAS) method to enhance the reactivity and recoverability of FeIII-TAML activators, which have been widely employed to degrade various organic contaminants. We have fabricated FeIII-TAML-based magnetic nanomaterials (FeIII-TAML/CTAB@Fe3O4, CTAB refers to cetyltrimethylammonium bromide) by adding a mixed solution of FeIII-TAML and NH3·H2O into another mixture containing CTAB, FeCl2 and FeCl3 solutions. The as-prepared FeIII-TAML/CTAB@Fe3O4 nanocomposite showed relative reactivity compared with free FeIII-TAML as indicated by decomposition of bisphenol A (BPA). Moreover, our results demonstrated that the FeIII-TAML/CTAB@Fe3O4 composite can be separated directly from reaction solutions by magnet adsorption and reused for at least four times. Therefore, the efficiency and recyclability of self-assembled FeIII-TAML/CTAB@Fe3O4 nanostructures will enable the application of FeIII-TAML-based materials with a lowered expense for environmental implication.
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Compuestos de Bencidrilo , Compuestos Férricos , Nanoestructuras , Fenoles , Compuestos Férricos/química , Cetrimonio , Fenómenos MagnéticosRESUMEN
INTRODUCTION: To assess the efficacy and safety of retrocorneal membrane interception (RMI)-enhanced penetrating canaloplasty in glaucoma secondary to iridocorneal endothelial syndrome (GS-ICE) with open angle or small peripheral anterior synechiae (PAS). METHODS: A series of 13 patients (13 eyes) with GS-ICE and uncontrolled intraocular pressure (IOP) underwent RMI-enhanced penetrating canaloplasty from March 2019 to October 2020. The patients were followed up at one week, 1 month, 3 months, 6 months, and 12 months, postoperatively. The main outcome measure was surgical success, which was defined as an IOP ≤ 18 mmHg or ≤ 21 mmHg, with glaucoma medications (qualified success), or without glaucoma medications (complete success). Secondary outcome measures were IOP, medication usage, and surgical complications. RESULTS: Among the 13 GS-ICE eyes, 12 (92%) achieved qualified success and 10 (77%) achieved complete success at 12 months postoperatively. The mean IOP decreased from 36.41 ± 8.92 mmHg on 3.0 (IQR 0.5) medications before surgery to 17.09 ± 7.71 mmHg (p = 0.0004) on 0.0 (IQR 0.5) medications (p = 0.0004) at 12 months after surgery. Transient hypotony (38%) and hyphema (38%) were the most common complications. CONCLUSIONS: RMI-enhanced penetrating canaloplasty appears effective and safe in treating GS-ICE with open angle or small PAS up to one year of follow up.
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Glaucoma de Ángulo Abierto , Presión Intraocular , Síndrome Endotelial Iridocorneal , Humanos , Glaucoma de Ángulo Abierto/cirugía , Glaucoma de Ángulo Abierto/fisiopatología , Glaucoma de Ángulo Abierto/complicaciones , Masculino , Femenino , Presión Intraocular/fisiología , Persona de Mediana Edad , Anciano , Síndrome Endotelial Iridocorneal/cirugía , Síndrome Endotelial Iridocorneal/diagnóstico , Síndrome Endotelial Iridocorneal/complicaciones , Estudios Retrospectivos , Estudios de Seguimiento , Agudeza Visual , Cirugía Filtrante/métodos , Resultado del Tratamiento , AdultoRESUMEN
BACKGROUND: There is a close correlation between HPV infection and systemic immune status. The purpose of this study was to determine which lymphocytes in peripheral blood influence human papillomavirus (HPV) infection and to identify whether peripheral blood lymphocyte (PBL) subsets could be used as biomarkers to predict HPV clearance in the short term. METHODS: This study involved 716 women undergoing colposcopy from 2019 to 2021. Logistic and Cox regression were used to analyze the association of PBLs with HPV infection and clearance. Using Cox regression, bidirectional stepwise regression and the Akaike information criterion (AIC), lymphocyte prediction models were developed, with the C-index assessing performance. ROC analysis determined optimal cutoff values, and their accuracy for HPV clearance risk stratification was evaluated via KaplanâMeier and time-dependent ROC. Bootstrap resampling validated the model and cutoff values. RESULTS: Lower CD4 + T cells were associated with a higher risk of HPV, high-risk HPV, HPV18 and HPV52 infections, with corresponding ORs (95% CI) of 1.58 (1.16-2.15), 1.71 (1.23-2.36), 2.37 (1.12-5.02), and 3.67 (1.78-7.54), respectively. PBL subsets mainly affect the natural clearance of HPV, but their impact on postoperative HPV outcomes is not significant (P > 0.05). Lower T-cell and CD8 + T-cell counts, as well as a higher NK cell count, are unfavorable factors for natural HPV clearance (P < 0.05). The optimal cutoff values determined by the PBL prognostic model (T-cell percentage: 67.39%, NK cell percentage: 22.65%, CD8 + T-cell model risk score: 0.95) can effectively divide the population into high-risk and low-risk groups, accurately predicting the natural clearance of HPV. After internal validation with bootstrap resampling, the above conclusions still hold. CONCLUSIONS: CD4 + T cells were important determinants of HPV infection. T cells, NK cells, and CD8 + T cells can serve as potential biomarkers for predicting natural HPV clearance, which can aid in patient risk stratification, individualized treatment, and follow-up management.
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Infecciones por Papillomavirus , Humanos , Femenino , Virus del Papiloma Humano , Estudios Retrospectivos , Linfocitos T CD4-Positivos , BiomarcadoresRESUMEN
Estrogen-related receptor alpha (ERRα) plays an important role in endometrial cancer (EC) progression. However, the biological roles of ERRα in EC invasion and metastasis are not clear. This study aimed to investigate the role of ERRα and 3-hydroxy-3-methylglutaryl-CoA synthase 1 (HMGCS1) in regulating intracellular cholesterol metabolism to promote EC progression. ERRα and HMGCS1 interactions were detected by co-immunoprecipitation, and the effects of ERRα/HMGCS1 on the metastasis of EC were investigated by wound-healing and transwell chamber invasion assays. Cellular cholesterol content was measured to verify the relationship between ERRα and cellular cholesterol metabolism. Additionally, immunohistochemistry was performed to confirm that ERRα and HMGCS1 were related to EC progression. Furthermore, the mechanism was investigated using loss-of-function and gain-of-function assays or treatment with simvastatin. High expression levels of ERRα and HMGCS1 promoted intracellular cholesterol metabolism for invadopodia formation. Moreover, inhibiting ERRα and HMGCS1 expression significantly weakened the malignant progression of EC in vitro and in vivo. Our functional analysis showed that ERRα promoted EC invasion and metastasis through the HMGCS1-mediated intracellular cholesterol metabolism pathway, which was dependent on the epithelial-mesenchymal transition pathway. Our findings suggest that ERRα and HMGCS1 are potential targets to suppress EC progression.
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Neoplasias Endometriales , Podosomas , Femenino , Humanos , Línea Celular Tumoral , Neoplasias Endometriales/patología , Hidroximetilglutaril-CoA Sintasa , Podosomas/fisiología , Receptores de Estrógenos/metabolismo , Invasividad Neoplásica , Metástasis de la Neoplasia , Receptor Relacionado con Estrógeno ERRalfaRESUMEN
An ultrasensitive 1O2-based electrochemical aptasensor by on-line assembly of photosensitizers using graphene oxide (GO) as a cartridge is reported. In the presence of target protein lysozyme, the interaction of lysozyme with aptamer led to the dissociation of dsDNA and release of the aptamer-lysozyme complex to solution, with DNA-c retaining on the electrode; then, the photosensitizer phloxine B (PB) was assembled on the electrode since GO can simultaneously adsorb DNA-c and PB molecules. Upon irradiation by a green LED, 1O2 was generated by photocatalysis of PB molecules and then cleaved the DNA-c, leading to remarkably decreased impedance signals that linearly respond with the logarithm of lysozyme concentration. Benefitting from the efficient photosensitization ability of PB and the high PB-loading capacity of GO, the developed sensor allowed determination of 0.001 to 100 nM lysozyme with a limit of detection of about 0.14 pM. The relative standard deviation (RSD) for five independent electrodes with 1 nM lysozyme was 3.1%, indicating satisfactory reproducibility. The sensor also showed excellent selectivity toward lysozyme in the presence of interfering substances and was applied to the determination of lysozyme in urine samples with recoveries ranging from 91 to 101%. The on-line assembly of photosensitizer technique opens a new way for amplified electrosensing of biomolecules. Graphical abstract An on-line assembly of photosensitizers and DNA on electrode was developed using graphene oxide a cartridge and the photocatalytic electrosensor can be used for label-free detection of lysozyme as low as 1 pM.
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A novel Gram-stain positive, short rod, forming sub-terminal endospores of ellipsoidal shape, halophilic, alkaliphilic and aerobic bacterium, designated strain KQ-12T, was isolated from a saline-alkaline lake in China, and characterised by a polyphasic taxonomic approach. The isolate grew at 4-40 °C (optimum, 25 °C), at pH 8.0-10.0 (pH 9.0) and in the presence of 0-16% (w/v) NaCl (8%). 16S rRNA gene sequence similarity of KQ-12T to species in the genera Salipaludibacillus ranged from 96.6 to 98.1%. Phylogenetic trees indicated that the strain should be assigned to the genus Salipaludibacillus. The polar lipids of KQ-12T were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and an unidentified phospholipid and its major cellular fatty acids were anteiso-C15:0, anteiso-C17:0, iso-C15:0, and C16:0. The isoprenoid quinone was MK-7. These key chemotaxonomic properties also confirmed the affiliation of the strain to the genus Salipaludibacillus. However, some physiological, biochemical properties, low average nucleotide identity and low digital DNA-DNA hybridization relatedness values enabled the strain to be differentiated from closely related species of the genus Salipaludibacillus. Thus, KQ-12T can be classified as a novel species in the genus Salipaludibacillus, for which the name Salipaludibacillus keqinensis sp. nov. is proposed. The type strain is KQ-12T (â= âACCC 60430Tââ = âKCTC 33935T).
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Bacillaceae/aislamiento & purificación , Lagos/microbiología , Bacillaceae/clasificación , Bacillaceae/genética , Bacillaceae/metabolismo , Técnicas de Tipificación Bacteriana , China , ADN Bacteriano/genética , Ácidos Grasos/química , Ácidos Grasos/metabolismo , Lagos/análisis , Filogenia , ARN Ribosómico 16S/genética , Cloruro de Sodio/análisis , Cloruro de Sodio/metabolismoRESUMEN
Exploration of quantum dots (QDs) as energy acceptors revolutionizes the current chemiluminescence resonance energy transfer (CRET), since QDs possess large Stokes shifts and high luminescence efficiency. However, the strong and high concentration of oxidant (typically H2O2) needed for luminol chemiluminescence (CL) reaction could cause oxidative quenching to QDs, thereby decreasing the CRET performance. Here we proposed the use of bienzyme-QDs bioconjugate as the energy acceptor for improved CRET sensing. Two enzymes, one for H2O2 generation (oxidase) and another for H2O2 consumption (horseradish peroxidase, HRP), were bioconjugated onto the surface of QDs. The bienzyme allowed fast in situ cascaded H2O2 generation and consumption, thus alleviating fluorescence quenching of QDs. The nanosized QDs accommodate the two enzymes in a nanometric range, and the CL reaction was confined on the surface of QDs accordingly, thereby amplifying the CL reaction rate and improving CRET efficiency. As a result, CRET efficiency of 30-38% was obtained; the highest CRET efficiency by far was obtained using QDs as the energy acceptor. The proposed CRET system could be explored for ultrasensitive sensing of various oxidase substrates (here exemplified with cholesterol, glucose, and benzylamine), allowing for quantitative measurement of a spectrum of metabolites with high sensitivity and specificity. Limits of detection (LOD, 3σ) for cholesterol, glucose, and benzylamine were found to be 0.8, 3.4, and 10 nM, respectively. Furthermore, multiparametric blood analysis (glucose and cholesterol) is demonstrated.
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Técnicas Biosensibles/métodos , Enzimas Inmovilizadas/química , Peróxido de Hidrógeno/análisis , Mediciones Luminiscentes/métodos , Luminol/química , Puntos Cuánticos/química , Animales , Bencilamino Oxidasa/química , Bencilaminas/análisis , Glucemia/análisis , Bovinos , Colesterol/sangre , Colesterol Oxidasa/química , Transferencia de Energía , Glucosa Oxidasa/química , Peroxidasa de Rábano Silvestre/química , Humanos , Modelos MolecularesRESUMEN
Culturable animal caliciviruses are widely-used as surrogates for human norovirus (HuNoV). The infectivity of a culturable virus was traditionally determined by plaque assay and/or 50% tissue culture infectious dose (TCID50) assay, both of which are time-consuming and labor-intensive. Molecular approaches, such as quantitative real time RT-PCR (qRT-PCR) and RT-PCR, could be used for detection of the viral genome but yet fail to determine the infectivity of a virus. In this study, we evaluated different assays for determination of infectivity of Tulane virus (TV), a surrogate for HuNoV. The infectivity of TV was measured by RNase exposure assay, RT-PCR assays, cellular-receptor-mediated capture qRT-PCR assay, receptor-mediated in situ capture qRT-PCR assay, cell-culture-mediated amplification qRT-PCR, and confirmed by TCID50 assay. RNase exposure assay was only useful for measuring TV inactivation caused by heat. Short template RT-PCR assay did not reflect inactivation status of TV. Partial reduction in viral RNA signal could be measured by long-template RT-PCR only when TV was inactivated by thermal or chlorine treatments at full-inactivation levels. Cellular-receptor-mediated capture qRT-PCR exhibited low sensitivity and specificity for the evaluation of virus infectivity. The in situ capture qRT-PCR assay could be used to evaluate virus inactivation deriving from damage to viral capsid caused by heat and chlorine. The cell-culture-mediated amplification qRT-PCR could be used as an alternative method to rapidly determine the infectivity of TV.
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Infecciones por Caliciviridae/virología , Caliciviridae/fisiología , Norovirus/fisiología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Caliciviridae/crecimiento & desarrollo , Línea Celular , Humanos , Viabilidad Microbiana , Norovirus/genética , Norovirus/crecimiento & desarrollo , Inactivación de VirusRESUMEN
Human noroviruses (HuNoVs) are the major cause of epidemic nonbacterial gastroenteritis. Although quantitative (real-time) reverse transcription-PCR (qRT-PCR) is widely used for detecting HuNoVs, it only detects the presence of viral RNA and does not indicate viral infectivity. Human blood group antigens (HBGAs) have been identified as receptors/co-receptors for both HuNoVs and Tulane virus (TV) and are crucial for viral infection. We propose that viral infectivity can be evaluated with a molecular assay based on receptor-captured viruses. In this study, we employed TV as an HuNoV surrogate to validate the HBGA-based capture qRT-PCR method against the 50% tissue culture infectious dose (TCID50) method. We employed type B HBGA on an immuno-well module to concentrate TV, followed by amplification of the captured viral genome by in situ qRT-PCR. We first demonstrated that this in situ capture qRT-PCR (ISC-qRT-PCR) method could effectively concentrate and detect TV. We then treated TV under either partial or full inactivation conditions and measured the remaining infectivity by ISC-qRT-PCR and a tissue culture-based amplification method (TCID50). We found that the ISC-qRT-PCR method could be used to evaluate virus inactivation deriving from damage to the capsid and study interactions between the capsid and viral receptor. Heat, chlorine, and ethanol treatment primarily affect the capsid structure, which in turns affects the ability of the capsid to bind to viral receptors. Inactivation of the virus by these methods could be reflected by the ISC-qRT-PCR method and confirmed by TCID50 assay. However, the loss of the infectivity caused by damage to the viral genome (such as that from UV irradiation) could not be effectively reflected by this method. Despite this limitation, the ISC-qRT-PCR provides an alternative approach to determine inactivation of Tulane virus. A particular advantage of the ISC-qRT-PCR method is that it is also a faster and easier method to effectively recover and detect the viruses, as there is no need to extract viral RNA or to transfer the captured virus from magnetic beads to PCR tubes for further amplification. Therefore, ISC-qRT-PCR can be easily adapted for use in automated systems for multiple samples.
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Caliciviridae/fisiología , Viabilidad Microbiana , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Virología/métodos , Inactivación de Virus , Antígenos de Grupos Sanguíneos/metabolismo , Caliciviridae/efectos de los fármacos , Caliciviridae/efectos de la radiación , Desinfección/métodos , Humanos , Receptores Virales/metabolismo , Acoplamiento ViralRESUMEN
In this work, we demonstrated a simple and highly sensitive LED-induced chemiluminescence (LED-CL) platform for aptasensing of proteins. It was based on the fact that fluorescein isothiocyanate (FITC) strongly catalyzed the reaction between luminol and dissolved oxygen under LED irradiation. Using FITC as a new catalytic tag, a highly sensitive sandwich type LED-CL aptasensor was fabricated for quantification of the model protein (platelet-derived growth factor-BB, PDGF-BB). Under optimized conditions, the assay for PDGF-BB was linear from 0.1 nM to 100 nM, and the detection limit was as low as 50 pM, which is better than most of other aptamer-based assays without an amplification procedure. Thrombin, bovine serum albumin, hemoglobin, cytochrome C and adenosine triphosphate showed no obvious interferences for the determination of PDGF-BB. The advantages of high sensitivity and selectivity provided by this LED-CL aptasensor will facilitate its wide application in protein assays.
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Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Sustancias Luminiscentes/química , Mediciones Luminiscentes/métodos , Factor de Crecimiento Derivado de Plaquetas/análisis , Animales , BovinosRESUMEN
Primary endometrial squamous cell carcinoma (PESCC) is a rare malignant tumor. To investigate the clinical and pathological features of PESCC, two cases of PESCC in Fujian Maternal and Child Health Hospital were retrospectively studied and the literatures were reviewed. Both of the two cases were menopausal women aged 57-62 years, clinically presenting with "vaginal discharge". Case 1 was a non-keratinising squamous cell carcinoma with high-risk HPV infection. Tumor infiltrated in deep myometrium with multifocal intravascular thrombus and macro metastases to one pelvic lymph node (1/15) and abdominal aortic lymph node (1/1). Lung metastasis occurred 36 months after the surgery. After surgical resection and without postoperative supplemental therapy, the patient remained tumor-free for 110 months to date. Case 2 had a history of breast cancer for 5 years and long-term intake of aromatase inhibitor drugs without HPV infection. It was a keratinized squamous cell carcinoma. Tumor also infiltrated in deep myometrium with multifocal intravascular thrombus and one pelvic lymph node metastasis (1/18), However, no metastasis was seen elsewhere. To date, the patient survived for 16 months without tumor after surgery. Both of the two cases expressed squamous epithelial markers P40, P63, and CK5/6, but neither expressed PAX8 or PR. Case 1 had diffuse expression of P16, wild-type P53, and ER-negative. Case 2 had negative P16, mutant P53, and focal positive ER. PESCC is often associated with HPV infection and low estrogen levels. However, studies in the literatures have found that P16 expression is not always consistent with HPV infection, indicating that PESCC cannot be easily classified as HPV-associated or non-dependent like cervical cancer. There are two main patterns of P16 and P53 expression, P16-positive/P53 wild-type and P16-negative/P53-mutant, but no positive expression of both has been seen so far. It is worth noting that we reported the second case of PESCC with a history of breast cancer, where the patient had been taking the oral aromatase inhibitor drug (exemestane) for a long period of time to reduce the estrogen level, indicating the low estrogen level may be also a key factor in the pathogenesis of PESCC.
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Objectives: Acute angle-closure glaucoma (AACG) is a major cause of irreversible and severe visual function loss. Robust rise in AACG was observed in the ophthalmic outpatient clinics concomitant with the outbreak of COVID-19 infection in China after the relaxing of "zero-COVID policy" in early December 2022. Here we compared the demographic and clinical proï¬les of patients with AACG before, during and after the COVID-19 outbreak. Underlying mechanisms were tentatively investigated. Methods: This is a retrospective comparative study. Consecutive cases with newly diagnosed AACGs in a tertiary eye hospital were retrospectively collected during December 17, 2022 to January 8, 2023. Data from the same period in 2018-2019 and 2019-2020, 2020-2021 and 2021-2022, and 2023-2024 were collected as pre-pandemic, pandemic-control, and post-pandemic control, respectively. For the patients in 2022-2023 outbreak group, COVID-19 infection was confirmed by reversed transcriptase-polymerase chain reaction or antibody test for severe acute respiratory syndrome coronavirus disease 2 (SARS-CoV-2) from nasopharyngeal swabs. Ocular parameters, serum electrolytes and coagulative parameters were compared between COVID-19 positive and negative AACGs in observational group. SARS-CoV-2 nucleic acid in the aqueous humor was detected. Results: A total of 106 AACG cases were diagnosed during the outbreak period in 2022-2023. In contrast, 18 (in 2018-2019) and 22 (in 2019-2020) cases were included during pre-pandemic period, and 21 (in 2023-2024) during the post-pandemic period. Only 13 and 4 newly onset AACG were included in 2020-2021 and 2021-2022 during the pandemic-control period, respectively. Younger age and higher proportion of bilateral involvement were detected in COVID-19 outbreak group than that of other groups (p = 0.034 and p = 0.080). Sixty-eight (64.2 %) patients in the outbreak group had a confirmed COVID-19 infection. Intervals between infection and AACG attack was 52 ± 85h (0-15d). Fifty-three patients (77.9 %) reported the applications of ibuprofen or other antipyretic medications and 25 (36.8 %) reported large volume water intake before AACG attack. COVID-19-positive AACG patients had higher level of D-dimer than their negative counterparts (1.13 ± 2.60 mg/L vs. 0.46 ± 0.43 mg/L, p = 0.083). No difference in IOP, serum electrolytes, and coagulative parameters other than D-dimer was observed between COVID-19 positive and negative cases. SARS-CoV-2 were negative in the aqueous humor from 14 COVID-19 positive and 8 negative patients. Conclusion: COVID-19 infection surged the onset of AACG in patients at risk. Mental stress, water intake, increased choroidal thickness due to SARS-CoV-2 induced ACE receptor activation, and hyper-coagulation, may contribute to the disease onset. Ocular involvement should not be ignored in both routine and new systemic emergent conditions.
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Transposon mutagenesis screening of Bacillus subtilis YB-1471, a novel rhizosphere biocontrol agent of Fusarium crown rot (FCR) of wheat, resulted in the identification of orf04391, linked to reduced biofilm formation. The gene encodes a protein possessing a putative tertiary structure of a "double-wing" DNA-binding domain. Expression of orf04391 increased during biofilm development in stationary cultures and during rapid growth in shaking cultures. An orf04391 deletion strain showed reduced biofilm production related to lower levels of the extracellular matrix, and the mutant also had reduced sporulation, adhesion, root colonization, and FCR biocontrol efficiency. Transcriptome analysis of YB-1471 and Δorf04391 in stationary culture showed that the loss of orf04391 resulted in altered expression of numerous genes, including sinI, an initiator of biofilm formation. DNA binding was shown with his-tagged Orf04391 binding to the sinIR operon in vivo and in vitro. Orf04391 appears to be a transcriptional regulator of biofilm formation in B. subtilis through the Spo0A-SinI/SinR pathway.
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Bacillus subtilis , Proteínas Bacterianas , Fusarium , Enfermedades de las Plantas , Triticum , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/química , Biopelículas/crecimiento & desarrollo , Fusarium/genética , Fusarium/metabolismo , Fusarium/fisiología , Regulación Bacteriana de la Expresión Génica , Enfermedades de las Plantas/microbiología , Triticum/microbiologíaRESUMEN
BACKGROUND: The evaluation of particle-bound mercury (PBM) exposure is a crucial aspect of assessing the global cycle of mercury (Hg) and its adverse effects on human health and ecosystems. Nevertheless, the precise and reliable measurement of PBM remains a formidable task because of the costly and cumbersome equipment required, as well as the inadequate sensitivities exhibited by current analytical techniques. In this study, we provided a unique and straightforward approach utilising filter fiber-assisted matrix solid-phase dispersion (FF-MSPD) in conjunction with single-drop solution electrode discharge-induced cold vapor generation atomic fluorescence spectrometry (SD-SEGD-CVG-AFS) for the precise quantification of PBM. The PBM contained in a small filter was efficiently extracted with 200 µL of eluent (0.2 % L-cysteine and 4 % HCOOH) by FF-MSPD and subsequently converted to Hg0 using SD-SEGD-CVG, before being subjected to examination using AFS. RESULTS: The resulted limit of detection (LOD, 3σ) was 0.17 pg m-3, obtained with a sample volume of 12 m3, which was much higher than that of the techniques published in the literatures. The aforementioned technique was effectively utilised for the detection of mercury in 19 samples of PM2.5 and PM10 which were collected over a span of several months. SIGNIFFCANCE: Contrast to conventional methods, the proposed method offers a range of distinct advantages, including simplified operation, absence of memory effects, enhanced sensitivity, substantial reduction in reagent usage, and decreased secondary pollution. These advantages are particularly valuable for advancing research on the fate, transport, and exposure routes of environmental mercury.
RESUMEN
This study reports the development of a photocatalytic electrochemical aptasensor for the purpose of detecting chloramphenicol (CAP) antibiotic residues in water by utilizing SYBR Green I (SG) and chemically exfoliated MoS2 (ce-MoS2) as synergistically signal-amplification platforms. The Au nanoparticles (AuNPs) were electrodeposited onto the surface of an indium tin oxide (ITO) electrode. After that, the thiolate-modified cDNA, also known as capture DNA, was combined with the aptamer. Subsequently, photosensitized SG molecules and ce-MoS2 nanomaterial were inserted into the groove of the resultant double-stranded DNA (dsDNA). The activation of the photocatalytic process upon exposure to light resulted in the generation of singlet oxygen. The singlet oxygen effectively split the dsDNA, resulting in significant enhancement in the current of [Fe(CN)6]3-/4-. When the CAP was present, both SG molecules and ce-MoS2 broke away from the dsDNA, which turned off the photosensitization response, leading to significant reduction in the current of [Fe(CN)6]3-/4-. Under the optimal conditions, the aptasensor exhibited a linear relationship between the current of [Fe(CN)6]3-/4- with logarithmic concentrations of CAP from 20 to 1000 nM, with a detection of limit (3σ) of 3.391 nM. The aptasensor also demonstrated good selectivity towards CAP in the presence of interfering antibiotics, such as tetracycline, streptomycin, levofloxacin, ciprofloxacin, and sulfadimethoxine. Additionally, the results obtained from the analysis of natural water samples using the proposed aptasensor were consistent with the findings acquired through the use of a liquid chromatograph-mass spectrometer. Therefore, with its simplicity and high selectivity, this aptasensor can potentially detect alternative antibiotics in environmental water samples by replacing the aptamers based on photosensitization.
Asunto(s)
Aptámeros de Nucleótidos , Benzotiazoles , Técnicas Biosensibles , Cloranfenicol , Diaminas , Disulfuros , Técnicas Electroquímicas , Molibdeno , Compuestos Orgánicos , Quinolinas , Cloranfenicol/análisis , Aptámeros de Nucleótidos/química , Técnicas Electroquímicas/métodos , Molibdeno/química , Diaminas/química , Disulfuros/química , Benzotiazoles/química , Quinolinas/química , Compuestos Orgánicos/química , Técnicas Biosensibles/métodos , Nanopartículas del Metal/química , Oro/química , Fármacos Fotosensibilizantes/química , Antibacterianos/análisis , Límite de Detección , Contaminantes Químicos del Agua/análisis , Procesos Fotoquímicos , Tamaño de la PartículaRESUMEN
Objective: This study introduced a novel subtype classification method for endometrial cancer (EC) with mismatch repair deficiency (MMRd) by employing immune status and prognosis as the foundational criteria. The goal was to enhance treatment guidance through precise subtype delineation. Methods: Study Cohort: This study encompassed a cohort of 119 patients diagnosed with MMRd-EC between 2015 and 2022. Analyses using t-tests and Mann-Whitney U-tests were performed to assess prognostic markers and peripheral blood immune cell profiles in patients with MutS deficiency (MutS-d) versus those with MutL deficiency (MutL-d). Logistic regression analysis was used to identify independent risk factors. Bioinformatics Analysis: An online database was used to assess the prognostic implications, immune cell infiltration, and immune checkpoint involvement associated with the deficiency of MutS versus MutL in EC. Results: Patients with MutL-d exhibited heightened risk factors, including elevated cancer grade and increased myometrial invasion, leading to poorer prognosis and shorter overall survival and progression-free survival. Regarding systemic immune status, patients with MutL-d demonstrated decreased peripheral blood lymphocyte percentage, lymphocyte count, and CD8+ T cell percentage. For local immunity, the infiltration of natural killer cells, CD8+ T cells, and cytotoxic T lymphocytes in the tumor tissue was reduced in patients with MutL-d. Additionally, patients with MutL-d exhibited lower expression of immune checkpoint markers. The composition of immune subtypes and survival outcomes also indicate that patients with MutL-d have a poorer immune status and prognosis than the patients with MutS-d. Conclusion: Patients with MMRd-EC can be subclassified according to MutS or MutL deficiency. Patients with MutS-d exhibited better immune status, prognosis, and immunotherapy benefits than those with MutL-d. These results can help guide patients to a more precise treatment.