RESUMEN
Bioorthogonal drug molecules are currently gaining prominence for their excellent efficacy, safety and metabolic stability. Pharmacokinetic study is critical for understanding their mechanisms and guiding pharmacotherapy, which is primarily performed with liquid chromatography-mass spectrometry as the gold standard. For broader and more efficient applications in clinics and fundamental research, further advancements are especially desired in cheap and portable instrumentation as well as rapid and tractable pretreatment procedures. Surface-enhanced Raman spectroscopy (SERS) is capable of label-free detection of various molecules based on the spectral signatures with high sensitivity even down to a single-molecule level. But limited by irreproducibility at low concentrations and spectral interference in complex biofluids, SERS hasn't been widely applied for pharmacokinetics, especially in live animals. In this work, we propose a new method to quantify bioorthogonal drug molecules with signatures at the spectral silent region (SR) by the digital colloid-enhanced Raman spectroscopy (dCERS) technique. This method was first validated using 4-mercaptobenzonitrile in a mixture of analogous molecules, exhibiting reliable and specific identification capability based on the unique SR signature and Poisson-determined quantification accuracy. We further developed a single-step serum pretreatment method and successfully profiled the pharmacokinetic behavior of an anticancer drug, erlotinib, from animal studies. In a word, this method, superior in sensitivity, controllable accuracy, minimal background interference and facile pretreatment and measurement, promises diverse applications in fundamental studies and clinical tests of bioorthogonal drug molecules.
RESUMEN
P2Y12 receptor inhibitors are commonly used in clinical antiplatelet therapy, typically alongside other medications. Vicagrel, a promising P2Y12 receptor inhibitor, has submitted a new drug marketing application to the United States Food and Drug Administration. Its primary metabolites and some metabolic pathways are identical to those of clopidogrel. The aim of this study was to investigate the effects of the thiol methyltransferase inhibitor (±)-2,3-dichloro-α-methylbenzylamine (DCMB) on the metabolism and pharmacokinetics of vicagrel. In vitro incubation with human and rat liver microsomes revealed that DCMB significantly inhibited the methylation of vicagrel's thiol metabolite M15-1. Rats were orally administered 6 mg/kg [14C]vicagrel (100 µCi/kg) 1 hour after peritoneal injection with or without DCMB (80 mg/kg). Compared with the control group, the plasma of DCMB-pretreated rats exhibited maximum plasma concentration (C max) decrease and time to reach C max (T max) delay for all vicagrel-related substances, the methylation product of the thiol metabolite (M9-2), and the derivatization product of the active thiol metabolite (MP-M15-2). However, no significant changes in area under the curve (AUC) or half-life (t 1/2) were observed. DCMB had negligible effect on the total radiological recovery of vicagrel within 72 hours, although the rate of vicagrel excretion slowed down within 48 hours. DCMB had a negligible impact on the metabolic pathway of vicagrel. Overall, the present study found that DCMB did not significantly affect the total exposure, metabolic pathways, metabolite profiles, or total excretion rates of vicagrel-related metabolites in rats, but led to C max decrease, T max delay, and slower excretion rate within 48 hours. SIGNIFICANCE STATEMENT: This study used liquid chromatography-tandem mass spectrometry combined with radiolabeling technology to investigate the effects of the thiol methyltransferase inhibitor (±)-2,3-dichloro-α-methylbenzylamine on the absorption, metabolism, and excretion of vicagrel in rats. This work helps to better understand the in vivo metabolism of active thiol metabolites of P2Y12 inhibitors such as clopidogrel, vicagrel, etc.
Asunto(s)
Metiltransferasas , Microsomas Hepáticos , Ratas Sprague-Dawley , Animales , Ratas , Masculino , Microsomas Hepáticos/metabolismo , Microsomas Hepáticos/efectos de los fármacos , Humanos , Metiltransferasas/antagonistas & inhibidores , Metiltransferasas/metabolismo , Bencilaminas/farmacocinética , Bencilaminas/farmacología , Metilación , Inhibidores Enzimáticos/farmacología , Inhibidores Enzimáticos/farmacocinética , Tiofenos/farmacocinética , Tiofenos/farmacología , Interacciones Farmacológicas , FenilacetatosRESUMEN
Triple-negative breast cancer (TNBC) represents a highly aggressive and heterogeneous malignancy. Currently, effective therapies for TNBC are very limited and remain a significant unmet clinical need. Targeting the transcription-regulating cyclin-dependent kinase 9 (CDK9) has emerged as a promising avenue for therapeutic treatment of TNBC. Herein, we report the design, synthesis, optimization, and evaluation of a new series of aminopyrazolotriazine compounds as orally bioavailable, potent, and CDK9/2 selectivity-improved inhibitors, enabling efficacious inhibition of TNBC cell growth, as well as notable antitumor effect in TNBC models. The compound C35 demonstrated low-nanomolar potency with substantially improved CDK9/2 selectivity, downregulated the CDK9-downstream targets (e.g., MCL-1), and induced apoptosis in TNBC cell lines. Moreover, with the desired oral bioavailability, oral administration of C35 could significantly suppress the tumor progression in two TNBC mouse models. This study demonstrates that target transcriptional regulation is an effective strategy and holds promising potential as a targeted therapy for the treatment of TNBC.
Asunto(s)
Antineoplásicos , Quinasa 9 Dependiente de la Ciclina , Inhibidores de Proteínas Quinasas , Neoplasias de la Mama Triple Negativas , Quinasa 9 Dependiente de la Ciclina/antagonistas & inhibidores , Quinasa 9 Dependiente de la Ciclina/metabolismo , Neoplasias de la Mama Triple Negativas/tratamiento farmacológico , Neoplasias de la Mama Triple Negativas/patología , Humanos , Animales , Femenino , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Antineoplásicos/farmacocinética , Antineoplásicos/química , Antineoplásicos/síntesis química , Administración Oral , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/uso terapéutico , Inhibidores de Proteínas Quinasas/farmacocinética , Inhibidores de Proteínas Quinasas/química , Inhibidores de Proteínas Quinasas/síntesis química , Ratones , Línea Celular Tumoral , Relación Estructura-Actividad , Disponibilidad Biológica , Proliferación Celular/efectos de los fármacos , Apoptosis/efectos de los fármacos , Descubrimiento de Drogas , Transcripción Genética/efectos de los fármacos , Ratones Desnudos , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Daratumumab, a humanized monoclonal antibody utilized in treating immunoglobulin light-chain amyloidosis and relapsed/refractory multiple myeloma, was quantified in rat serum through a simple, economical and effective liquid chromatography tandem-mass spectrometry (LC-MS/MS) method. A surrogate peptide, LLIYDASNR, derived from trypsin hydrolysis, was quantitatively analyzed with LLIYDASN [13C6, 15N4] RAT as an internal standard. This corrected variations from sample pretreatment and mass spectrometry response, involving denaturation and trypsin hydrolysis in a two-step process lasting approximately 1â¯hour. Methodological validation demonstrated a linear range of 1⯵g/mL to 1000⯵g/mL in rat serum. Precision, accuracy, matrix effect, sensitivity, stability, selectivity, carryover, and interference met acceptance criteria. The validated LC-MS/MS approach was successfully applied to a pharmacokinetic study of daratumumab in rats at an intravenous dose of 15â¯mg/kg.
Asunto(s)
Cromatografía Líquida con Espectrometría de Masas , Espectrometría de Masas en Tándem , Cromatografía Liquida/métodos , Tripsina , Espectrometría de Masas en Tándem/métodos , Anticuerpos Monoclonales/química , Inmunoglobulina G , Digestión , Reproducibilidad de los ResultadosRESUMEN
23-hydroxybetulinic acid (23-HA), a main bioactive component isolated from Pulsatilla chinensis (Bunge) Regel, exhibits various pharmacological activities, such as antimelanoma, antileukemia, anti-colon cancer, and antihepatotoxicity. Although the main active ingredient anemoside B4 (AB4) from this plant has been well studied, research on its active metabolite 23-HA is limited. In the present study, a validated HPLC-QQQ-MS/MS method was established for the quantification of 23-HA in rat plasma. Pharmacokinetics analysis showed that the absorption and elimination of 23-HA in rats were rapid, with an oral bioavailability as 12.9 %. After oral administration with 50 mg/kg 23-HA for SD rats, the plasma, urine, feces, and bile samples were collected and analyzed by UPLC-Q Exactive Plus MS and HPLC-QQQ-MS/MS. Seventeen metabolites of 23-HA were identified, and its major metabolic pathways included oxidation, hydration, sulfation, and glucuronidation. This study highlights the first detailed investigation of 23-HA's pharmacokinetics in rats along with its metabolism in vivo, and will provide robust evidence for further research and clinical application of 23-HA.
Asunto(s)
Cromatografía Líquida con Espectrometría de Masas , Espectrometría de Masas en Tándem , Triterpenos , Ratas , Animales , Ratas Sprague-Dawley , Cromatografía Líquida de Alta Presión/métodos , Espectrometría de Masas en Tándem/métodos , Heces/química , Administración OralRESUMEN
A new liquid chromatography tandem mass spectrometry (LC-MS/MS) method was established to quantify the anti-gastric cancer fully human monoclonal antibody (ramucirumab) in rat and human serum. The surrogate peptide (GPSVLPLAPSSK) for ramucirumab was generated by trypsin hydrolysis and quantified using the isotopically labeled peptide GPSVLPLAPSSK[13C6, 15N2]ST containing two more amino acids at the carboxyl end as an internal standard to correct for variations introduced during the enzymatic hydrolysis process and any mass spectrometry changes. Additionally, the oxidation and deamidation of unstable peptides (VVSVLTVLHQDWLNGK and NSLYLQMNSLR) were detected. The quantitative range of the proposed method was 1-1000 µg/mL, and complete methodological validation was performed. The precision, accuracy, matrix effect, sensitivity, stability, selectivity, carryover, and interference of the measurements met the required standards. The validated LC-MS/MS method was applied to pharmacokinetic studies in rats administered ramucirumab at 15 mg/kg intravenously. Overall, a robust, efficient, and cost-effective LC-MS/MS method was successfully developed for quantifying ramucirumab in rat and human serum.
Asunto(s)
Ramucirumab , Espectrometría de Masas en Tándem , Humanos , Ratas , Animales , Cromatografía Liquida/métodos , Espectrometría de Masas en Tándem/métodos , Cromatografía Líquida con Espectrometría de Masas , Péptidos/química , Inmunoensayo , Digestión , Reproducibilidad de los ResultadosRESUMEN
BACKGROUND: There has been no systematic review and meta-analysis to analyze and summarize the predictive factors of successful sperm extraction in salvage microdissection testicular sperm extraction. OBJECTIVES: We aimed to investigate the factors predicting the result of salvage microdissection testicular sperm extraction in patients with non-obstructive azoospermia who failed the initial microdissection testicular sperm extraction or conventional testicular sperm extraction. MATERIALS AND METHODS: We conducted a systematic literature search in PubMed, Web of Science, EMBASE, and the Cochrane Library for literature that described the characteristics of patients with non-obstructive azoospermia who underwent salvage microdissection testicular sperm extraction after failing the initial microdissection testicular sperm extraction or conventional testicular sperm extraction published prior to June 2022. RESULTS: This meta-analysis included four retrospective studies with 332 patients with non-obstructive azoospermia who underwent a failed initial microdissection testicular sperm extraction and three retrospective studies with 177 non-obstructive azoospermia patients who underwent a failed conventional testicular sperm extraction. The results were as follows: among non-obstructive azoospermia patients whose first surgery was microdissection testicular sperm extraction, younger patients (standard mean difference: -0.28, 95% confidence interval [CI]: -0.55 to -0.01) and those with smaller bilateral testicular volume (standard mean difference: -0.55, 95% CI: -0.95 to -0.15), lower levels of follicle-stimulating hormone (standard mean difference: -0.86, 95% CI: -1.18 to -0.54) and luteinizing hormone (standard mean difference: -0.68, 95% CI: -1.16 to -0.19), and whose testicular histological type was hypospermatogenesis (odds ratio: 3.52, 95% CI: 1.30-9.53) were more likely to retrieve spermatozoa successfully, while patients with Sertoli-cell-only syndrome (odds ratio: 0.41, 95% CI: 0.24-0.73) were more likely to fail again in salvage microdissection testicular sperm extraction. Additionally, in patients who underwent salvage microdissection testicular sperm extraction after a failed initial conventional testicular sperm extraction, those with testicular histological type of hypospermatogenesis (odds ratio: 30.35, 95% CI: 8.27-111.34) were more likely to be successful, while those with maturation arrest (odds ratio: 0.39, 95% CI: 0.18-0.83) rarely benefited. CONCLUSION: We found that age, testicular volume, follicle-stimulating hormone, luteinizing hormone, hypospermatogenesis, Sertoli-cell-only syndrome, and maturation arrest were valuable predictors of salvage microdissection testicular sperm extraction, which will assist andrologists in clinical decision-making and minimize unnecessary injury to patients.
Asunto(s)
Azoospermia , Oligospermia , Síndrome de Sólo Células de Sertoli , Humanos , Masculino , Azoospermia/cirugía , Azoospermia/patología , Oligospermia/patología , Estudios Retrospectivos , Microdisección/métodos , Recuperación de la Esperma , Semen , Testículo/cirugía , Testículo/patología , Espermatozoides/patología , Hormona Folículo Estimulante , Hormona Luteinizante , Hormona Folículo Estimulante HumanaRESUMEN
Recombinant human growth hormone (rhGH) is a peptide comprising 191 amino acids, that is mainly used to promote the growth of children and plays an important antiaging role. In the present study, a simple and sensitive quantitation method for rhGH in rat plasma was established by LCMS/MS. After simple and rapid enzymatic digestion of the plasma sample, two suitable surrogate peptides (LFDNAMLR and FPTIPLSR) were selected for quantitative analysis. The results showed good linearity over calibration range 10-2000 ng/mL. The quality control (QC) accuracy ranged from -13.8 to 14.3%, and the accuracy of the lower limit of quantification (LLOQ) ranged from -12.9 to 19.0%. The intra-day and inter-day precision ranges for all QCs were 1.7-13.6% and 4.0-7.0%, respectively. The method was successfully applied to intravenous and subcutaneous pharmacokinetic studies in rats. In comparison with previously published methods, our method features simple sample preparation combined with a short sample processing time (3.5 h), wide linear range (10-2000 ng/mL), small plasma volume (35 µL), and LLOQ (10 ng/mL).
Asunto(s)
Hormona de Crecimiento Humana , Animales , Humanos , Ratas , Cromatografía Liquida/métodos , Hormona de Crecimiento Humana/análisis , Hormona de Crecimiento Humana/sangre , Control de Calidad , Reproducibilidad de los Resultados , Espectrometría de Masas en Tándem/métodos , Proteínas Recombinantes/análisis , Proteínas Recombinantes/sangreRESUMEN
In this paper, α-alanine and ß-alanine were used as modifiers to prepare α-alanine amidated pectin (α-AP) and ß-alanine amidated pectin (ß-AP) through enzymatic method. The effects of alanine and its isomer on the amidation degree and physicochemical properties of pectin were studied. Fourier transform infrared spectrum, proton nuclear magnetic resonance analysis, X-ray photoelectron spectrum and elemental analysis indicated that the amino groups from α-C and ß-C could be aminated with the carboxyl group of pectin to form the corresponding aminated pectin. The alanine grafting ratio of α-AP and ß-AP were 21.99% and 18.92%, respectively. The results showed that the dynamic viscosity of ß-AP was significantly higher than that of α-AP due to the influence of amino acid side chain. However, due to the higher alanine grafting ratio of α-AP, the strength of hydrogel prepared by α-AP was higher than that of ß-AP, and it also has the highest elastic modulus and swelling capacity. In addition, pectin, as a carbohydrate-based fat substitute, has been widely used in the field of food to simulate the smooth and delicate taste of fat. Compared with High methoxy pectin (HMP) and ß-AP, α-AP had better colloid stability and smaller hydrogel particles in the composite system. The results provide references for the application of amidated pectin in low-fat food.
Asunto(s)
Sustitutos de Grasa , Pectinas , Alanina , Hidrogeles , Pectinas/química , beta-AlaninaRESUMEN
In this study, pectin was modified with phenylalanine by ultra-low temperature enzymatic method to improve its gel properties. The grafting ratio of phenylalanine amidated pectin was studied under different reaction conditions. The highest value (29.21 %) was reached a reaction temperature of -5 °C and time of 12 h. Further analysis indicated that phenylalanine and high methoxyl pectin combined at the solid-liquid two phase interface under the catalysis of papain to form phenylalanine amidated pectin. Moreover, the physicochemical properties of pectin hydrogel and its feasibility as a sustained-release drug carrier were discussed. The results showed that phenylalanine amidated pectin can form hydrogel with a certain strength under acidic conditions, and there is no need to add a lot of soluble solids and divalent cations. Besides, the phenylalanine amidated pectin hydrogel as a sustained release carrier of drugs showed more sustained and complete drug release.
Asunto(s)
Hidrogeles , Pectinas , Preparaciones de Acción Retardada , Liberación de Fármacos , Hidrogeles/química , Pectinas/química , Fenilalanina , TemperaturaRESUMEN
OBJECTIVE: Accurate prediction of postoperative recurrence risk of gastric cancer (GC) is critical for individualized precision therapy. We aimed to investigate whether a computed tomography (CT)-based radiomics nomogram can be used as a tool for predicting the local recurrence (LR) of GC after radical resection. MATERIALS AND METHODS: 342 patients (194 in the training cohort, 78 in the internal validation cohort, and 70 in the external validation cohort) with pathologically proven GC from two centers were included. Radiomics features were extracted from the preoperative CT imaging. The clinical model, radiomics signature, and radiomics nomogram, which incorporated the radiomics signature and independent clinical risk factors, were developed and verified. Furthermore, the performance of these three models was assessed by using the area under the curve (AUC) of receiver operating characteristic (ROC) curve analysis and decision curve analysis (DCA). RESULTS: The radiomics signature, which was comprised of two selected radiomics features, namely, contrast_GLCM and dissimilarity_GLCM, showed better performance than the clinical model in predicting the LR of GC, with AUC values of 0.83 in the training cohort, 0.84 in the internal validation cohort, and 0.73 in the external cohort, respectively. By integrating the independent clinical risk factors (N stage, bile acid duodenogastric reflux and nodular or irregular outer layer of the gastric wall) into the radiomics signature, the radiomics nomogram achieved the highest accuracy in predicting LR, with AUC values of 0.89, 0.89 and 0.80 in the three cohorts, respectively. DCA in the validation cohort showed that radiomics nomogram added more net benefit than the clinical model within the range of 0.01-0.98. CONCLUSION: The CT-based radiomics nomogram has the potential to predict the LR of GC after radical resection.
RESUMEN
The physiological process of male reproduction relies on the orchestration of neuroendocrine, immune, and energy metabolism. Spermatogenesis is controlled by the hypothalamic-pituitary-testicular (HPT) axis, which modulates the production of gonadal steroid hormones in the testes. The immune cells and cytokines in testes provide a protective microenvironment for the development and maturation of germ cells. The metabolic cellular responses and processes in testes provide energy production and biosynthetic precursors to regulate germ cell development and control testicular immunity and inflammation. The metabolism of immune cells is crucial for both inflammatory and anti-inflammatory responses, which supposes to affect the spermatogenesis in testes. In this review, the role of immunometabolism in male reproduction will be highlighted. Obesity, metabolic dysfunction, such as type 2 diabetes mellitus, are well documented to impact male fertility; thus, their impacts on the immune cells distributed in testes will also be discussed. Finally, the potential significance of the medicine targeting the specific metabolic intermediates or immune metabolism checkpoints to improve male reproduction will also be reassessed.
Asunto(s)
Metabolismo Energético , Inmunomodulación , Reproducción/fisiología , Animales , Manejo de la Enfermedad , Susceptibilidad a Enfermedades , Retroalimentación Fisiológica , Hormonas Esteroides Gonadales/metabolismo , Hormona Liberadora de Gonadotropina/metabolismo , Humanos , Sistema Hipotálamo-Hipofisario/inmunología , Sistema Hipotálamo-Hipofisario/metabolismo , Sistema Inmunológico/inmunología , Sistema Inmunológico/metabolismo , Infertilidad Masculina/etiología , Infertilidad Masculina/metabolismo , Infertilidad Masculina/terapia , Masculino , Testículo/inmunología , Testículo/metabolismoRESUMEN
OBJECTIVE: To evaluate the relationship between the BRAF V600E mutation in lymph node metastasis (LNM) and its invasive characteristics in papillary thyroid cancer (PTC). MATERIAL AND METHODS: A total of 373 PTC patients were enrolled in this study conducted at Zhujiang Hospital of Southern Medical University between January 2017 and December 2018. PTCs with cervical lymph node metastases were verified pathohistologically, and primary tumors and LNM were examined for the BRAF V600E mutation. Patients were excluded from the study if the BRAF V600E mutation was examined only in primary tumors or only in LNM. RESULTS: Of the 373 patients examined, BRAF V600E mutation frequency in primary tumors was slightly higher than in LNM (81.5% vs 78.0%, P = 0.000), the intra-class correlation coefficient (ICC) was 0.865 (95% CI 0.835-0.890). The BRAF V600E mutation in both primary tumor and LNM negatively correlated with the size of the largest metastatic focus of LNM (Odds ratio, OR = 0.297, 95% CI 0.143-0.616, P = 0.001; OR = 0.242, 95% CI 0.119-0.492, P = 0.000, respectively). There was no relationship between BRAF V600E mutation in LNM and the number, extranodal extension or stage of LNM (P > 0.05). CONCLUSION: The BRAF V600E mutation in LNM may not be related to the invasive characteristics of LNM in PTC.
RESUMEN
The synthesized sheet-like polypyrrole (ppy) nanowires were used as solid phase extraction materials, followed by gas chromatography-mass spectrometry (GC-MS) for the detection of traces residues of pyrethroid pesticides in human plasma. A multiresidue method was developed and verified for the determination of trace pyrethroid residues (transfluthrin, allethrin, resmethrin, fenpropathrin, etofenprox, fenvalerate) in human plasma. In this study, using the cationic surfactant cetyltrimethylammonium bromide (CTAB) as a soft template, ppy nanowires with regular morphology were prepared by oxidative polymerization. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction analysis (XRD), Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA) and other techniques were employed for characterization. Molecular dynamics analyses were used to simulate the adsorption mechanism of each pyrethroid and ppy nanowires. Based on density analysis, molecular recognition analysis, and binding energy, the van der Waals force was considered as an important driving force for the adsorption of pyrethroids and ppy nanowires. The limits of detection (LOD) of six pyrethroids were 0.008-0051 ng mL-1, and the limits of quantification (LOQ) were 0.028-0.162 ng mL-1. The relative standard deviations of ppy nanowires were 2.12-5.09%, and the recoveries of six pyrethroids ranged from 76.9 to 110.4%. The enrichment factors were within the range of 47.09-51.30. The experimental results showed that the method could be an efficient detection method for trace residue analysis of pyrethroid pesticides in complex biological samples. It would be advantageous for clinical monitoring and toxicological studies of pyrethroids.
Asunto(s)
Simulación de Dinámica Molecular , Nanocables/química , Plaguicidas/sangre , Polímeros/química , Piretrinas/sangre , Pirroles/química , Microextracción en Fase Sólida/métodos , Adsorción , Cromatografía de Gases y Espectrometría de Masas , Humanos , Enlace de Hidrógeno , Iones , Reproducibilidad de los Resultados , Espectroscopía Infrarroja por Transformada de Fourier , Termodinámica , Termogravimetría , Factores de Tiempo , Difracción de Rayos XRESUMEN
The vast majority of first-trimester pregnancy losses are the consequence of numerical aberrations in fetal chromosomes, which may involve nearly all chromosomes. Although commercial probes for all chromosomes are available for multiplex ligation-dependent probe amplification (MLPA) and fluorescence in situ hybridization (FISH) analyses, their use has rarely been reported for screening all 24 chromosomes for early fetal demise, especially by FISH. Here, we validated the ability of MLPA and FISH techniques as two low-cost aneuploidy screening methods for 24 chromosomes in 165 chorionic villus samples (CVSs). The results obtained by two methods were compared by the Chi-square test and the Kappa agreement test. Both methods gave conclusive results for all CVSs tested and showed highly consistent results (kappaâ¯=â¯0.890, pâ¯<â¯0.001). There was no statistically significant difference between the aneuploidy rate of the CVSs tested by the two methods (pâ¯=â¯0.180). Most of the samples showed fully concordant molecular karyotyping results (81.21%) between the two analytical methods, 10.91% had incompletely concordant results, and 7.88% had discordant results. The inconsistencies included segmental abnormalities, mosaicism, and polyploidy. Both assays used to screen 24 chromosomes were powerful techniques for detecting aneuploidy in CVSs. In terms of cost-effectiveness and diagnostic accuracy, the combination of subtelomeric (P036, P070) and centromeric (P181) MLPA assays is the better analytic strategy and follow-up analysis by FISH is recommended for MLPA-negative samples.
Asunto(s)
Aneuploidia , Muestra de la Vellosidad Coriónica/métodos , Vellosidades Coriónicas/fisiopatología , Hibridación Fluorescente in Situ , Análisis Citogenético , Femenino , Humanos , Mosaicismo , Reacción en Cadena de la Polimerasa Multiplex , EmbarazoRESUMEN
Lung cancer is the leading cause of cancer death worldwide, and non-small cell lung cancer (NSCLC) accounts for 80%-85% of diagnostic cases. The molecular mechanisms of NSCLC pathogenesis are not well understood. Heterogeneous nuclear ribonucleoprotein K (hnRNPK) is a multifunctional protein that regulates gene expression and signal transduction and closely associated with tumorigenesis, but its mechanism of action in the pathogenesis of NSCLC is unclear. In this study, we observed that the expression pattern of hnRNPK in H1299 lung adenocarcinoma cells varied depending on the cell density in culture. Moreover, hnRNPK stimulated the ability of proliferation and colony formation of H1299 cells, which is important for the multilayered cell growth in culture. We further investigated whether there is an association between hnRNPK and the elements involved in the cell contact inhibition pathway. By using quantitative reverse transcriptase-polymerase chain reaction assay and a YAP activity reporter system, we found that hnRNPK upregulated the mRNA and protein levels and transcriptional activity of Yes-associated protein 1 (YAP), a master negative regulator of Hippo contact inhibition pathway. Furthermore, YAP knockdown with siRNA abolished the stimulatory effect of hnRNPK on H1299 cell proliferation. These results suggested that YAP could be one of the effectors of hnRNPK. Our data may provide new clues for further understanding the biological functions of hnRNPK, particularly in the context of lung adenocarcinoma oncogenesis.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Adenocarcinoma del Pulmón/genética , Carcinoma de Pulmón de Células no Pequeñas/genética , Regulación Neoplásica de la Expresión Génica , Ribonucleoproteína Heterogénea-Nuclear Grupo K/genética , Neoplasias Pulmonares/genética , Factores de Transcripción/genética , Células A549 , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Adenocarcinoma del Pulmón/metabolismo , Adenocarcinoma del Pulmón/patología , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Línea Celular , Línea Celular Tumoral , Proliferación Celular/genética , Supervivencia Celular/genética , Ribonucleoproteína Heterogénea-Nuclear Grupo K/metabolismo , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Interferencia de ARN , Transducción de Señal/genética , Factores de Transcripción/metabolismo , Proteínas Señalizadoras YAPRESUMEN
The inflammatory response is an unavoidable process and contributes to the destruction of cerebral tissue during the acute ischemic stroke (AIS) phase and has not been addressed fully to date. Insightful understanding of correlation of inflammatory mediators and stroke outcome may provide new biomarkers or therapeutic approaches for ischemic stroke. Here, we prospectively recruited 180 first-ever AIS patients within 72 hrs after stroke onset. We used the National Institutes of Health Stroke Scale (NIHSS) to quantify stroke severity and modified Rankin scale (mRS) to assess the 3-month outcome for AIS patients. Initially, we screened 35 cytokines, chemokines, and growth factors in sera from 75 AIS patients and control subjects. Cytokines that were of interest were further investigated in the 180 AIS patients and 14 heathy controls. We found that IL-1RA, IL-1ß, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-13, IL-15, EGF, G-CSF, Flt-3L, GM-CSF and Fractalkine levels were significantly decreased in severe stroke patients. In particular, IL-1ß, IL-4, IL-5, IL-7, IL-9, IL-10, IL-15, G-CSF and GM-CSF were significantly reduced in AIS patients with poor outcome, compared to those with good prognosis. IL-6 was notably higher in the poor outcome group. Only IL-9 level decreased in the large infarct volume group. After adjusting for confounders, we found that IL-5 was an independent protective factor for prognosis in AIS patients with an adjusted OR of 0.042 (P = 0.007), whereas IL-6 was an independent risk predictor for AIS patients with an adjusted OR of 1.293 (P = 0.003). Our study suggests the levels of serum cytokines are related to stroke severity, short-term prognosis and cerebral infarct volume in AIS patients.
RESUMEN
BACKGROUND: Although a number of studies have reported the role of an increased left atrial (LA) size on stroke, limited data are collected about the relationship between LA enlargement and recurrent ischemic stroke in the Chinese population. Our aim was to assess the association of LA size with the risk of stroke recurrence, particularly with recurrent cardioembolic or cryptogenic stroke in ischemic stroke patients. METHODS: The study recruited 313 consecutive patients with acute first-ever ischemic stroke. Echocardiographic LA diameter was measured and indexed by height and body surface area separately. The endpoint was recurrent ischemic stroke. Cox proportional hazard models were used to examine the association of LA size with total recurrent ischemic stroke and recurrent cardioembolic or cryptogenic stroke while adjusting for baseline demographics characteristics, clinical factors, echocardiographic left ventricular ejection fraction, and medication. RESULTS: Over a median follow-up period of 1.63 years, 47 recurrent ischemic strokes (21 were cardioembolic or cryptogenic) occurred. In a multivariate model adjusted for potential confounders, compared with the bottom tertiles of LA diameter indexed to height (LA diameter/H), the top tertile of LA diameter/H was significantly associated with the total recurrent ischemic stroke (adjusted HR 3.610, 95% CI 1.870-6.967, p < .001) and the composite of recurrent cardioembolic or cryptogenic stroke (adjusted HR 5.673, 95% CI 1.780-18.084, p = .003). Results were similar when LA diameter indexed to body surface area (LA diameter/BSA) was involved in the analysis. CONCLUSION: LA size is an independent predictor of total recurrent ischemic stroke and the composite of recurrent cardioembolic or cryptogenic stroke.
Asunto(s)
Isquemia Encefálica/complicaciones , Atrios Cardíacos , Accidente Cerebrovascular/epidemiología , Anciano , China/epidemiología , Ecocardiografía/métodos , Femenino , Atrios Cardíacos/diagnóstico por imagen , Atrios Cardíacos/patología , Humanos , Masculino , Persona de Mediana Edad , Tamaño de los Órganos , Modelos de Riesgos Proporcionales , Recurrencia , Factores de Riesgo , Accidente Cerebrovascular/etiologíaRESUMEN
BACKGROUND: Serum soluble lectin-like oxidized low-density lipoprotein receptor-1 (sLOX-1) has been shown associated with the progression of atherosclerosis in endothelial cells. We sought to assess whether the baseline serum sLOX-1 levels are correlated with the presence and short-term functional outcome of large-artery atherosclerotic (LAA) stroke. METHODS: The study recruited 241 subjects, including 148 consecutive patients with acute ischemic stroke with the subtype of LAA and 93 non-stroke controls. Clinical and laboratory data, including serum concentration of sLOX-1, were collected within 24 h of admission, and the severity of LAA stroke patients was evaluated by National Institutes of Health Stroke Scale score. And functional outcome was assessed by modified Rankin Scale three months after stroke. The association between sLOX-1 level and the functional outcome at three months was analyzed by multiple logistic regression models. RESULTS: Serum levels of sLOX-1 in the LAA stroke patients were significantly higher as compared to normal controls (2.48 ± 0.93 ng/ml vs. 2.22 ± 0.79 ng/ml in the controls, t = 2.301, p = 0.022). The levels of serum sLOX-1 in patients with good outcome were significantly lower than those with poor outcome (2.39 ± 0.94 ng/ml vs. 2.77 ± 0.84 ng/ml, p = 0.032). After adjusting for potential confounders, sLOX-1 was still an independent predictor for the function outcome with an adjusted OR of 3.39 (95% CI, 1.61-7.11, p = 0.001). CONCLUSIONS: The serum sLOX-1 level was higher in patients with LAA stroke, and it was an independent predictor of functional outcome in patients with LAA ischemic stroke.
Asunto(s)
Isquemia Encefálica/diagnóstico , Receptores Depuradores de Clase E/sangre , Accidente Cerebrovascular/diagnóstico , Anciano , Biomarcadores/sangre , Isquemia Encefálica/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Índice de Severidad de la Enfermedad , Accidente Cerebrovascular/sangreRESUMEN
BACKGROUND: Neutrophil-to-lymphocyte ratio is an independent predictor of mortality in patients with acute ischemic stroke. However, it is uncertain whether neutrophil-to-lymphocyte ratio is related with functional outcome and recurrent ischemic stroke. In this study, we aimed to investigate the relationship of neutrophil-to-lymphocyte ratio with stroke severity, functional outcome, and recurrent ischemic stroke after acute ischemic stroke. METHODS: A total of 280 patients with acute ischemic stroke were included in the study. Patients were divided into 3 groups according to the neutrophil-to-lymphocyte ratio value (<2, 2-3, >3). Demographic, clinical, and laboratory data were collected for all patients. We evaluated the association between neutrophil-to-lymphocyte ratio and (1) stroke severity on admission, (2) functional outcome at 3 months, and (3) recurrent ischemic stroke. Regression analyses were performed, adjusting for confounders. RESULTS: After adjustment for potential confounders, neutrophil-to-lymphocyte ratio was associated with an increased risk of stroke severity on admission (odds ratio [OR] 1.364, 95% confidence interval [CI] 1.101-1.690, P = .005) and primary unfavorable outcome (OR 1.455, 95% CI 1.083-1.956, P = .013). After a median of 1.13 years (interquartile range.91-1.42) of follow-up, neutrophil-to-lymphocyte ratio was associated with recurrent ischemic stroke after adjustment (hazard ratio 1.499, 95% CI 1.161-1.935, P = .002). CONCLUSIONS: Our study suggests that neutrophil-to-lymphocyte ratio is associated with stroke severity on admission, primary unfavorable functional outcome, and recurrent ischemic stroke in patients with acute ischemic stroke.