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In this work, bright yellow fluorescent and multifunctional carbon dots (N-CDs) were prepared by hydrothermal method from O-phenylenediamine and 4-aminobenzoic acid. The fluorescence characterization showed that the N-CDs possessed good optical properties (QY = 32%) and excitation dependent multi-color emission. By exciting with 390 nm, the strong selective interaction of VB12 with N-CDs could result in a sharp decrease in the luminescence of N-CDs at 567 nm. An efficient fluorescence sensor in aqueous solution was constructed which could linearly respond VB12 in wide concentration ranges of 0-90 µM and 140-250 µM. The linear correlation coefficients of N-CDs and VB12 were 0.9950 and 0.9968, respectively, and the detection limit was 0.119 µM. N-CDs were performed for sensitive determination of VB12 in real samples. Moreover, the N-CDs were exploited to image cell. This N-CDs was a sensitive fluorescence probe to monitor VB12 and presented prospective potential in living cells imaging. Schematic diagram of the synthesis process and application research of N-CDs.
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In this paper, intrinsic dual-emission fluorescent carbon dots (CDs) doped with N and S atoms have been firstly fabricated. The characterization results show that CDs are successfully synthesized with two separate fluorescence emissions at 468 nm and 628 nm, respectively. The strong and selective interaction of Cr (VI) ions with CDs lead to obvious fluorescence decrease of CDs at 468 nm, which is caused by a mixed quenching mechanism. At the same time, the fluorescence at 628 nm increase. Interestingly, the CDs solution show obvious color change under the daylight and UV light, so visualization detection of Cr (VI) can be realized in water samples. Based on the data of the emission intensity ratios of F468/F628, Cr (VI) can be detected from 3.8 to 38.9 µM combined with the linear correlation coefficient of 0.998, and the lowest detection concentration is 47.2 nM. The platform is satisfactorily applied to the detection of Cr (VI) ions in water samples. In addition, the CDs could be applied as fluorescent probes for cell imaging with dual fluorescent emission.
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Carbono , Pontos Quânticos , Corantes Fluorescentes , Íons , ÁguaRESUMO
We assessed the concordance between immunohistochemistry (IHC) and gene expression profiling (GEP) for determining diffuse large B-cell lymphoma (DLBCL) cell of origin (COO) in the phase III PHOENIX trial of rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP) with or without ibrutinib. Among 910 of 1114 screened patients with non-germinal centre B cell-like (non-GCB) DLBCL by IHC, the concordance with GEP for non-GCB calls was 82·7%, with 691 (75·9%) identified as activated B cell-like (ABC), and 62 (6·8%) as unclassified. Among 746 of 837 enrolled patients with verified non-GCB DLBCL by IHC, the concordance with GEP was 82·8%, with 567 (76·0%) identified as ABC and 51 (6·8%) unclassified; survival outcomes were similar regardless of COO or treatment, whereas among patients with ABC DLBCL aged <60 years, the overall and event-free survival were substantially better with ibrutinib versus placebo plus R-CHOP [hazard ratio (HR) 0·365, 95% confidence interval (CI) 0·147-0·909, P = 0·0305; HR 0·561, 95% CI 0·326-0·967, P = 0·0348, respectively]. IHC and GEP showed high concordance and consistent survival outcomes among tested patients, indicating centralised IHC may be used to enrich populations for response to ibrutinib plus R-CHOP.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Perfilação da Expressão Gênica , Imuno-Histoquímica , Linfoma Difuso de Grandes Células B/classificação , Adenina/administração & dosagem , Adenina/análogos & derivados , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Linfócitos B/química , Linfócitos B/patologia , Ciclofosfamida/administração & dosagem , Doxorrubicina/administração & dosagem , Centro Germinativo/patologia , Humanos , Estimativa de Kaplan-Meier , Linfoma Difuso de Grandes Células B/tratamento farmacológico , Linfoma Difuso de Grandes Células B/genética , Linfoma Difuso de Grandes Células B/patologia , Pessoa de Meia-Idade , Células-Tronco Neoplásicas/química , Células-Tronco Neoplásicas/patologia , Piperidinas/administração & dosagem , Prednisona/administração & dosagem , Prognóstico , Intervalo Livre de Progressão , Rituximab/administração & dosagem , Resultado do Tratamento , Vincristina/administração & dosagemRESUMO
BACKGROUND: It has been reported that lifestyle factors may affect birth weight; however, few studies have explored the association between lifestyle factors and low birth weight in preterm and term births in China. The objective of this study was to explore the effect of lifestyle on low birth weight in preterm and term births. METHODS: This case-control study was conducted in fourteen hospitals in Jiangmen, Guangdong Province. Data were collected from August 2015 to May 2016 using a standard questionnaire. Data were analysed using logistic regression. RESULTS: Women who delivered preterm and were physically active (1-3 times per week and ≥ 4 times per week) had reduced odds of having low birth weight babies (aOR = 0.584, 95%CI = 0.394-0.867 and, aOR = 0.516, 95%CI = 0.355-0.752, respectively). Pregnant women who had insufficient gestational weight gain had increased odds of having low birth weight babies (aOR = 2.272, 95%CI = 1.626-3.176). Women exposed to passive smoking had an increased risk of delivering low birth weight infants (aOR = 1.404, 95%CI = 1.057-1.864). Insufficient gestational weight gain and excessive gestational weight gain were both significantly associated with low birth weight (aOR = 1.484, 95%CI = 1.103-1.998 and aOR = 0.369, 95%CI = 0.236-0.577, respectively) for term deliveries. In addition, parity, history of low birth weight, antenatal care and gestational hypertension were significantly associated with the likelihood of low birth weight. CONCLUSION: Pregnant women without exercise contraindications should remain physically active. Pregnant women should be aware of the negative effects of smoke and be aware of strategies to protect themselves from passive smoke exposure. Hospitals should inform pregnant women of the importance appropriate gestational weight gain. These recommendations should be put into practice to decrease the prevalence of low birth weight infants.
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Recém-Nascido de Baixo Peso , Estilo de Vida , Comportamento Materno/fisiologia , Nascimento Prematuro/epidemiologia , Nascimento a Termo , Estudos de Casos e Controles , China/epidemiologia , Exercício Físico , Feminino , Idade Gestacional , Humanos , Gravidez , Cuidado Pré-Natal/estatística & dados numéricos , Fatores de Risco , Fatores Socioeconômicos , Poluição por Fumaça de Tabaco , Aumento de PesoRESUMO
A fluoroimmunoassay based on quantum dots (QDs) and a lateral flow immunoassay system based on the magnetic beads (MB) were constructed to detect ractopamine (RAG) in urine samples. The monoclonal antibody (Ab1) against RAC was conjugated with QDs or MB as detector reagent, respectively. They apply a competitive format using an immobilized RAC conjugate and free RAC present in samples. That is to say, the concentration of RAC in the sample was negative related to the fluorescense intensity of QDs or the color density of MB. Results showed that the limit of detection (LOD) of fluorescence immunoassay method is 1 ng · mL⻹ and analysis time is 4 h, while the visual LOD was 10 ng · mL⻹ and analysis time was 15 min in magnetic lateral flow immunoassay system (MFLIS). Taken into consideration of the advantages and disadvantages of the two methods, it was suitable for the trace detection of RAC using fluoroimmunoassay while it was appropriate for point-of-care tesing of RAC by MFLIS.
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Fluorimunoensaio , Imunoensaio , Fenetilaminas/urina , Anticorpos Monoclonais/química , HumanosRESUMO
Fluoroimmunoassay based on quantum dots (QDs) and magnetic relaxation switch (MRS) immunoassay based on superparamagnetic nanoparticles (SMN) were constructed to detect Salmonella enterica (S. enterica) in water samples. In fluoroimmunoassay, magnetic beads was conjugated with S. enterica capture antibody (MB-Ab2) to enrich S. enterica from sample solution, then the QDs was conjugated with the S. enterica detection antibody (QDs-Ab1) to detect S. enterica based on sandwich immunoassay format. And the fluorescence intensity is positive related to the bacteria concentration of the sample. Results showed that the limit of detection (LOD) of this method was 102 cfu · mL⻹ and analysis time was 2 h. In MRS assay, magnetic nanoparticle-antibody conjugate (MN-Ab1) can switch their dispersed and aggregated state in the presence of the target. This state of change can modulate the spin-spin relaxation time (T2) of the neighboring water molecule. The change in T2(ΔT2) positively correlates with the amount of the target in the sample. Thus, AT can be used as a detection signal in MRS immunosensors. Results showed that LOD of MRS sensor for S. enterica was 10³ cfu · mL⻹ and analysis time was 0.5 h. Two methods were compared in terms of advantages and disadvantages in detecting S. enterica.
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Fluorimunoensaio , Salmonella enterica/isolamento & purificação , Água/análise , Fenômenos Magnéticos , Nanopartículas de Magnetita , Pontos Quânticos , Microbiologia da ÁguaRESUMO
Abusive use of ß-agonists as feed additives for animals and medication is detrimental to human health and food safety. Conventional assays are restricted to a single type of ß-agonists detection and cannot match the multiplexing features to perform automated, high throughput, and rapid quantitative analysis in real samples. In this research, we develop a portable automated chip system (PACS) with highly integrated automated devices in conjunction with portable microfluidic chips to provide simultaneous point-of-care testing of multiple ß-agonists in the field, simplifying complex manual methods, shortening assay times, and improving sensitivity. Specifically, silicon film is used as reaction substrates for immobilizing the conjugates of ß-agonists to increase the sensitivity of the assay result. Then, the PACS with a chemiluminescence imaging detector is established for automatic high-throughput and sensitive detection of Clenbuterol, Ractopamine, and Salbutamol based on the indirect immunoassay. Newly developed chip with high mixing performance can improve the sensitivity of target determination. Multiplex assays were carried out using the developed system for Clenbuterol, Ractopamine, and Salbutamol with a limit of detection of 54 pg mL-1,59 pg mL-1, and 93 pg mL-1, respectively. Except for sample preparation and coating, the detection in the PACS takes less than 47 min. A satisfactory sample recovery (86.33%-108.12%) was obtained, validating the reliability and practical applicability of this PACS. Meanwhile, the PACS enables sensitive and rapid detection of multiple ß-agonists in farms or markets where lacking advanced laboratory facilities.
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Técnicas Biossensoriais , Clembuterol , Animais , Humanos , Reprodutibilidade dos Testes , Albuterol , Testes ImediatosRESUMO
UNLABELLED: Study Type - Diagnostic (exploratory cohort) Level of Evidence 2b What's known on the subject? and What does the study add? Hypermethylation of genes such as glutathione-S-transferase P1 (GSTP1) and adenomatous polyposis coli (APC) occurs with high frequency in prostate tumour tissue but is much less common in the benign prostate; however, the potential value of gene methylation biomarkers as an adjunct to biopsy histopathology has had little study. When measured in histologically benign prostate biopsy tissue, APC gene hypermethylation was found to have high negative predictive value and high sensitivity. GSTP1 hypermethylation was found to have lower performance than APC. OBJECTIVE: To evaluate the performance of DNA methylation biomarkers in the setting of repeat biopsy in men with an initially negative prostate biopsy but a high index of suspicion for missed prostate cancer. PATIENTS AND METHODS: We prospectively evaluated 86 men with an initial histologically negative prostate biopsy and high-risk features. All men underwent repeat 12-core ultrasonography-guided biopsy. DNA methylation of glutathione-S-transferase P1 (GSTP1) and adenomatous polyposis coli (APC) was determined using tissue from the initially negative biopsy and compared with histology of the repeat biopsy. The primary outcome was the relative negative predictive value (NPV) of APC compared with GSTP1, and its 95% confidence interval (CI). RESULTS: On repeat biopsy, 21/86 (24%) men had prostate cancer. APC and GSTP1 methylation ratios below the threshold (predicting no cancer) produced a NPV of 0.96 and 0.80, respectively. The relative NPV was 1.2 (95% CI: 1.06-1.36), indicating APC has significantly higher NPV. Methylation ratios above the threshold yielded a sensitivity of 0.95 for APC and 0.43 for GSTP1. Combining both methylation markers produced a performance similar to that of APC alone. APC methylation patterns were consistent with a possible field effect or occurrence early in carcinogenesis. CONCLUSIONS: APC methylation provided a very high NPV with a low percentage of false-negatives, in the first prospective study to evaluate performance of DNA methylation markers in a clinical cohort of men undergoing repeat biopsy. The potential of APC methylation to reduce unnecessary repeat biopsies warrants validation in a larger prospective cohort.
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Biomarcadores Tumorais/genética , Biópsia por Agulha , Metilação de DNA , Genes APC , Glutationa S-Transferase pi/genética , Próstata/patologia , Neoplasias da Próstata/diagnóstico , Adulto , Idoso , Reações Falso-Negativas , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Sensibilidade e EspecificidadeRESUMO
Amivantamab, an epidermal growth factor receptor (EGFR)-c-Met bispecific antibody, targets activating/resistance EGFR mutations and MET mutations/amplifications. In the ongoing CHRYSALIS study (ClinicalTrials.gov Identifier: NCT02609776), amivantamab demonstrated antitumor activity in patients with non-small cell lung cancer harboring EGFR exon 20 insertion mutations (ex20ins) that progressed on or after platinum-based chemotherapy, a population in which amivantamab use has been approved by the US Food and Drug Administration. This bridging study clinically validated two novel candidate companion diagnostics (CDx) for use in detecting EGFR ex20ins in plasma and tumor tissue, Guardant360 CDx and Oncomine Dx Target Test (ODxT), respectively. From the 81 patients in the CHRYSALIS efficacy population, 78 plasma and 51 tissue samples were tested. Guardant360 CDx identified 62 positive (16 negative), and ODxT identified 39 positive (3 negative), samples with EGFR ex20ins. Baseline demographic and clinical characteristics were similar between the CHRYSALIS-, Guardant360 CDx-, and ODxT-identified populations. Agreement with local PCR/next-generation sequencing tests used for enrollment into CHRYSALIS demonstrated high adjusted negative (99.6% and 99.9%) and positive (100% for both) predictive values with the Guardant360 CDx and ODxT tests, respectively. Overall response rates were comparable between the CHRYSALIS, Guardant360 CDx, and ODxT populations. Both the plasma- and tissue-based diagnostic tests provided accurate, comprehensive, and complementary approaches to identifying patients with EGFR ex20ins who could benefit from amivantamab therapy.
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Carcinoma Pulmonar de Células não Pequenas , Neoplasias Pulmonares , Humanos , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/genética , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/tratamento farmacológico , Neoplasias Pulmonares/genética , Seleção de Pacientes , Mutagênese Insercional/genética , Inibidores de Proteínas Quinases/uso terapêutico , Receptores ErbB/genética , Éxons/genética , MutaçãoRESUMO
In this post hoc subgroup analysis of 200 patients enrolled in China from the phase III PHOENIX trial (N = 838, NCT01855750), addition of ibrutinib to rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisone (R-CHOP) did not improve event-free survival (EFS) versus placebo+R-CHOP in the intent-to-treat (ITT; n = 200, hazard ratio [HR] = 0.83, 95% confidence interval [CI]: 0·509-1.349; p = 0.4495) or activated B-cell-like (ABC; n = 141 [based on available gene-expression profiling data], HR = 0.86, 95% CI: 0.467-1.570; p = 0.6160) subpopulations. However, ibrutinib+R-CHOP improved EFS (HR = 0·50, 95% CI: 0.251-1.003) and progression-free survival (PFS; HR = 0.48, 95% CI: 0.228-1.009) versus placebo+R-CHOP in patients aged <60 but not ≥60 years. Grade ≥3 serious treatment-emergent adverse events occurred more with ibrutinib+R-CHOP (45·6% vs. 31·3%). The percentage of patients receiving ≥6 cycles of R-CHOP was similar across treatment arms in those <60 years. A numerical trend was seen towards improved EFS and PFS with ibrutinib+R-CHOP versus placebo+R-CHOP in patients with MYC-high/BCL2-high co-expression. In this slightly younger Chinese subgroup, ibrutinib+R-CHOP did not improve EFS in the ITT and ABC subpopulations but improved outcomes with manageable safety in patients <60 years, consistent with overall PHOENIX study outcomes.
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OBJECTIVE: Serial circulating tumor cell (CTC) counts have demonstrated predictive and prognostic value in patients with metastatic breast, colorectal, and prostate cancer. In a phase III study of pegylated liposomal doxorubicin (PLD) with trabectedin vs. PLD for relapsed ovarian cancer, we evaluated the correlation, if any, between numbers of CTCs and progression free survival, (PFS) and overall survival (OS). METHODS: CTCs were isolated from peripheral blood (10 mL) using the CellSearch system and reagents (Veridex). A CTC is defined as EpCAM+, cytokeratin+, CD45-, and is positive for the nuclear stain DAPI. The normal reference range for CellSearch is <2 CTC/7.5 mL of blood. Hazard ratios adjusted for known prognostic factors were estimated by Cox regression. RESULTS: Two-hundred sixteen patients had baseline CTC measurements of which 111 (51.4%) were randomized to the trabectedin+PLD arm; 143/216 patients (66.2%) were platinum-sensitive. Thirty-one of 216 patients (14.4%) had 2 or more CTCs detected prior to the start of therapy (range 2-566). Univariate Cox regression analyses indicated that patients with ≥2 CTCs prior to therapy had 1.89- (p=0.003) and 2.06-fold (p=0.003) higher risk for progression and death respectively. Multivariate analyses that include baseline CA-125, platinum sensitivity status, largest diameter lesion, number of tumor lesions, ECOG PS, and tumor grade show that patients with elevated baseline CTC had 1.58- (p=0.058) and 1.54-fold (p=0.096) higher risk for progression and death respectively. CONCLUSIONS: Results from this study indicate that elevated numbers of CTCs impart an unfavorable prognosis for ovarian cancer patients.
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Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Recidiva Local de Neoplasia/sangue , Recidiva Local de Neoplasia/tratamento farmacológico , Células Neoplásicas Circulantes/patologia , Neoplasias Ovarianas/sangue , Neoplasias Ovarianas/tratamento farmacológico , Dioxóis/administração & dosagem , Intervalo Livre de Doença , Doxorrubicina/administração & dosagem , Doxorrubicina/análogos & derivados , Feminino , Humanos , Pessoa de Meia-Idade , Análise Multivariada , Recidiva Local de Neoplasia/patologia , Neoplasias Ovarianas/patologia , Polietilenoglicóis/administração & dosagem , Taxa de Sobrevida , Tetra-Hidroisoquinolinas/administração & dosagem , TrabectedinaRESUMO
This study examined the effect of subcellular localization of P21 on the proliferation and apoptosis of HepG2 cells. The coding genes of the wild and the mutant P21 were amplified by mega primer PCR from the plasmid pCEP-WAF1 which contains human P21 cDNA in the nuclear localizational signal (NLS) sequence, and then inserted into the eukaryotic expression vector pDsRed1-C1. The recombinants were transfected into HepG2 cells. The transcription and expression of P21 were determined by RT-PCR and fluorescence microscopy. The cell proliferation was measured by MTT, and the cell cycle and apoptosis of HepG2 cells by flow cytometry. The results of restriction analysis, DNA sequencing and fluorescence microscopy confirmed the construction of the wild and the mutant P21 in the eukaryotic expression plasmid. The plasmid containing the mutant P21 was found to accelerate cell proliferation and the wild P21 plasmid to inhibit cell proliferation. Cell cycle analysis showed that the cell ratio of G(0)/G(1) in the wild type group was significantly increased as compared with that in the mutant type group, and cell apoptosis analysis revealed that the apoptosis rate in the wild type group was much higher than that in the mutant type group. It was concluded that the subcellular localization of P21 may contribute to the development of hepatic cancer.
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Apoptose/fisiologia , Proliferação de Células , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Citoplasma/metabolismo , Sequência de Bases , Núcleo Celular/metabolismo , Inibidor de Quinase Dependente de Ciclina p21/fisiologia , Células Hep G2 , Humanos , Dados de Sequência Molecular , Distribuição TecidualRESUMO
Conventional immunoassay methods have their common defects, such as tedious processing steps and inadequate sensitivity, in detecting whole blood. To overcome the above problems, we report a microfluidic chip-based magnetic relaxation switching (MRS) immunosensor via enzyme-mediated nanoparticles to simplify operation and amplify the signal in detecting whole blood samples. In the silver mirror reaction with catalase (CAT) as the catalyst, H2O2 can effectively control the production of Ag NPs. The amount of Ag NPs formed further affects the degree of aggregation of magnetic nanoparticles (MNPS), which gives rise to the changes of transverse relaxation time (T2). Both sample addition and reagent reaction are carried out in the microfluidic chip, thereby saving time and reagent consumption. We also successfully apply the sensor to detect alpha-fetoprotein (AFP) in real samples with a satisfied limit of detection (LOD = 0.56 ng/ml), which is superior to the conventional ELISA.
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Existing methods for detecting cardiac markers are difficult to be applied in point-of-care testing (POCT) due to complex operation, long time consumption, and low sensitivity. Here, we report a snail-shaped microfluidic chip (SMC) for the multiplex detection of cTnI, CK-MB, and Myo with high sensitivity and a short detection time. The SMC consists of a sandwich structure: a channel layer with a mixer and reaction zone, a reaction layer coated with capture antibodies, and a base layer. The opening or closing of the microchannels is realized by controlling the downward movement of the press-type mechanical valve. The chemiluminescence method was used as a signal readout, and the experimental conditions were optimized. SMC could detect cTnI, CK-MB, and Myo at concentrations as low as 1.02, 1.37, and 4.15. The SMC will be a promising platform for a simultaneous determination of multianalytes and shows a potential application in POCT.
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AIM: Women have an increased risk for developing depression during pregnancy, and depression has a serious negative impact on the mother and infant. This study explored the effectiveness and feasibility of a comprehensive intervention based on using a short message service (SMS) to reduce depressive symptoms and prevent depression during pregnancy. METHODS: This quasi-experimental study was conducted in three public hospitals with similar levels of care and maternal origin in Jiangmen City, Guangdong Province. One of the three hospitals was randomly selected as the intervention hospital, and the others were control hospitals. There were 4501 pregnant women who participated in this study. Pregnant women in the intervention group received a comprehensive intervention based on SMS after enrollment. Data were collected using questionnaires from August 2016 to August 2018. RESULTS: After the intervention, the Edinburgh Postnatal Depression Scale scores of the intervention group were lower than those of the control group (intervention group: 3.9 ± 3.9, control group: 5.2 ± 4.3, P < .001), and the proportion of subjects with positive depression screening results in the intervention group was also significantly lower than that in the control group (intervention group: 9.0%, control group: 16.1%, P < .001). Moreover, compared with women in the intervention group, women in the control group who did not receive the intervention were more likely to be positive for depression in the third trimester (AOR = 2.04, 95% CI = 1.62-2.58). CONCLUSIONS: The SMS-based comprehensive intervention used in this study can effectively alleviate depressive symptoms and reduce the risk of depression during pregnancy.
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Depressão Pós-Parto , Complicações na Gravidez , Envio de Mensagens de Texto , Depressão/diagnóstico , Depressão/prevenção & controle , Depressão Pós-Parto/prevenção & controle , Feminino , Humanos , Gravidez , Complicações na Gravidez/prevenção & controle , Gestantes , Escalas de Graduação PsiquiátricaRESUMO
Enhancing tumor homing and improving the efficacy of drugs are urgent needs for cancer treatment. Herein a novel targeted, intracellularly activatable fluorescence and cytotoxicity nanodiamond (ND) drug system (ND-PEG-HYD-FA/DOX, NPHF/D) was successfully prepared based on doxorubicin (DOX) and folate (FA) covalently bound to PEGylated NDs, in which the DOX was covalently coupled via an intracellularly hydrolyzable hydrazone bond that was stable in the physiological environment to ensure minimal drug release in circulation. Cell uptake studies demonstrated the selective internalization of NPHF/D by folate receptor (FR) mediated endocytosis in the order MCF-7 > HeLa > HepG2 â« CHO, using confocal laser scanning microscopy (CLSM) and flow cytometry. Interestingly, the DOX fluorescence of NPHF/D was significantly quenched, while the fluorescence recovery and cytotoxicity took place by low pH regulation in intracellular lysosomes, which made NPHF/D act as a fluorescence OFF-ON messenger for activatable imaging and cancer therapy. Of note, NPHF/D significantly inhibited the growth of tumors. Simultaneously, it was demonstrated that the introduction of FA and the cleavability of the hydrazone greatly enhanced the therapeutic performance of NPHF/D. In addition, toxicity studies in mice verified that the composites were devoid of any detected hepatotoxicity, cardiotoxicity, and nephrotoxicity using histopathology and blood biochemistry studies. Our work provides a novel strategy for cancer therapy, using ND-conjugated cancer drugs, and the exploration of theranostic drug-delivery systems.
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Antineoplásicos/química , Materiais Biocompatíveis/química , Portadores de Fármacos/química , Nanodiamantes/química , Animais , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Materiais Biocompatíveis/metabolismo , Materiais Biocompatíveis/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Doxorrubicina/química , Doxorrubicina/metabolismo , Doxorrubicina/farmacologia , Doxorrubicina/uso terapêutico , Liberação Controlada de Fármacos , Endocitose , Ácido Fólico/análogos & derivados , Ácido Fólico/química , Humanos , Concentração de Íons de Hidrogênio , Masculino , Camundongos , Camundongos Nus , Microscopia Confocal , Neoplasias/tratamento farmacológico , Neoplasias/patologia , Polietilenoglicóis/química , Nanomedicina Teranóstica , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Erdafitinib, a pan-fibroblast growth factor receptor (FGFR) inhibitor received accelerated approval from the US Food and Drug Administration (FDA) for locally advanced or metastatic urothelial carcinoma (mUC) in adult patients with specific FGFR3/2 genetic alterations who progressed during or after ≥1 line of prior platinum-containing chemotherapy (PCC), including within 12 months of neoadjuvant or adjuvant PCC. Concordance between the clinical trial assay (CTA) used in a phase 2 study and QIAGEN's therascreen® FGFR kit (a two-step, multiplex, real-time, RT-PCR assay), the FDA-approved companion diagnostic (CDx) with erdafitinib, was evaluated in this bridging study. Study samples included 100 CTA-confirmed FGFR-positive samples from 100 erdafitinib-treated mUC patients, plus 200 CTA-confirmed FGFR-negative samples from the phase 2 study. The primary objective was met if the lower bound of 95% CI of objective response rate (ORR) in CDx-confirmed patients with FGFR alterations was >25%. Demographics were similar between the bridging study and CTA-screened patients. In total, 292 of 300 samples (97.3%) with valid CDx results showed high analytical concordance versus CTA (percent agreement [95% CI]: positive percent agreement, 87.2 [79.0; 92.5]; negative percent agreement, 97.0 [93.5; 98.6]; overall percent agreement, 93.8 [90.5; 96.1]). Investigator-assessed ORR in the 81 CDx-identified, erdafitinib-treated patients who tested positive for both assays was 45.7% (95% CI: 35.3%; 56.5%) versus 40.4% (95% CI: 30.7%; 50.1%) for CTA and met the criteria for primary objective. High ORR and clinical concordance to CTA suggest that QIAGEN's CDx can reliably select mUC patients who would potentially benefit from erdafitinib treatment.
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Carcinoma de Células de Transição , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/genética , Receptor Tipo 3 de Fator de Crescimento de Fibroblastos/genética , Neoplasias Urológicas , Idoso , Carcinoma de Células de Transição/diagnóstico , Carcinoma de Células de Transição/tratamento farmacológico , Carcinoma de Células de Transição/genética , Carcinoma de Células de Transição/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pirazóis/uso terapêutico , Quinoxalinas/uso terapêutico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/antagonistas & inibidores , Receptor Tipo 3 de Fator de Crescimento de Fibroblastos/antagonistas & inibidores , Neoplasias Urológicas/diagnóstico , Neoplasias Urológicas/tratamento farmacológico , Neoplasias Urológicas/genética , Neoplasias Urológicas/patologiaRESUMO
BACKGROUND: Skeletal dysplasias account for nearly 10% of fetal structural malformations detected by ultrasonography. This clinically heterogeneous group of genetic anomaly includes at least 461 genetic skeletal disorders with extreme clinical, phenotypic, and genetic heterogeneities, thus, significantly complicates accurate diagnosis. Researches have used whole exome sequencing (WES) for prenatal molecular diagnoses of skeletal dysplasias, however, data are still limited. METHODS: DNA extracted from umbilical cord blood or amniocytes from fetuses suspected of skeletal dysplasias based on ultrasound evaluations were analyzed by WES. Blood samples were taken from the parents of the positive fetuses for co-segregation analysis using Sanger sequencing. RESULT: Definitive molecular diagnosis was made in 6/8 (75%) cases, comprised of 5 de novo disease-causing changes in 3 genes (FGFR3, COL2A1, and COL1A2) and one proband with a biallelic deficiency for Lamin B Receptor(LBR),and including 3 novel variants. All fetuses had no detectable copy number variation (CNV) from sequencing results. CONCLUSIONS: Our study suggests that WES is an efficient approach for prenatal diagnosis of fetuses suspected of skeletal abnormalities and contributes to parental genetics counseling and pregnancy management.
Assuntos
Sequenciamento do Exoma , Osteocondrodisplasias/genética , Diagnóstico Pré-Natal , Adulto , China , Feminino , Feto/anormalidades , Humanos , Osteocondrodisplasias/sangue , Ondas Ultrassônicas , Adulto JovemRESUMO
BACKGROUND: Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway plays an important role in sepsis, transducing a multitude of inflammatory signals. To date, knowledge of JAK/STAT pathway in sepsis is limited. This study was to investigate the potential role of JAK/STAT pathway in mediating multiple organ damage and mortality in septic rats. Our data showed that inhibition of JAK2/STAT3 attenuated cecal ligation and puncture-induced multiple organ damage and mortality in 48 hours in rats. METHODS: A total of 98 male Wistar rats were randomly divided into 4 groups as follows: (1) normal control group (n = 10); (2) cecal ligation and puncture (CLP) group (n = 40), which was further divided into 2, 6, 24, 48-hour post-CLP groups; (3) AG490 (8.0 mg/kg, Calbiochem, La Jolla, CA) treatment group (n = 24), which was further divided into 2, 6, 24, 48-hour post-CLP groups; (4) rapamycin (0.4 mg/kg, Calbiochem, Calbiochem, La Jolla, CA) treatment group (n = 24), which was further divided into 2, 6, 24, 48-hour post-CLP groups; CLP was performed to induce experimental sepsis. AG490 (8 mg/kg) or rapamycin (0.4 mg/kg) was injected subcutaneously 0.5 hour before CLP in respective group. Animals were killed at destined time after CLP (not including the death rate observation group), and specimens of serum, liver, and lungs were harvested and stored in liquid nitrogen for subsequent analyses. In an additional experiment, 88 animals were randomly divided into three groups to compare the survival rate, including CLP group (n = 40), AG490 treatment group (8 mg/kg, n = 24), and rapamycin treatment group (0.4 mg/kg, n = 24). Mortality of rats in each group was recorded up to 48 hours after the procedure. RESULTS: After CLP challenge, myeloperoxidase (MPO), aspartate transaminase, and alanine aminotransferase levels, as well as activation of JAK2 and STAT3, were markedly increased. Administration of AG490 or rapamycin significantly decreased activation of JAK2 and STAT3, as well as high mobility group box-1 protein, MPO, alanine aminotransferase levels (p < 0.05 or p < 0.01). In addition, treatment with AG490 or rapamycin significantly improved the 48-hour survival rate from 37.5% (15 of 40) to 66.7% (16 of 24) and 70.8% (17 of 24), respectively (both p < 0.05). CONCLUSION: JAK2/STAT3 pathway might play a role in the development of multiple organ damage in septic rats, which suggested a potential strategy to control sepsis.
Assuntos
Ceco/lesões , Inibidores Enzimáticos/farmacologia , Janus Quinase 2/antagonistas & inibidores , Janus Quinase 2/genética , Insuficiência de Múltiplos Órgãos/genética , Fator de Transcrição STAT3/antagonistas & inibidores , Fator de Transcrição STAT3/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Sirolimo/farmacologia , Tirfostinas/farmacologia , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases/sangue , Esquema de Medicação , Proteína HMGB1/sangue , Masculino , Insuficiência de Múltiplos Órgãos/mortalidade , Peroxidase/sangue , Ratos , Ratos Wistar , Taxa de SobrevidaRESUMO
OBJECTIVE: To investigate the potential role of Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway in regulation of gene expression of high mobility group box-1 protein (HMGB1) in various tissues in rats with sepsis. METHODS: A sepsis model reproduced by cecal ligation and puncture (CLP), and 128 male Wistar rats were randomly divided into normal control group (n = 10), sham operation group (n = 10), CLP group (n = 60), AG490 treatment group (n = 24), and rapamycin (RPM) treatment group (n = 24). At serial time points animals in each group were sacrificed after CLP, then tissue samples were harvested to determine HMGB1 mRNA expression and STAT1/3 DNA binding activity. RESULTS: STAT1 activities increased rapidly in the liver, lungs and small intestine after CLP, peaking at 6 - 12 h, while it increased slowly, and still kept at mild level from 2 to 48 h in the kidneys. Compared with STAT1, lower STAT3 activities were detected only in the liver and lungs, with negative detection in the small intestine and kidneys. HMGB1 mRNA levels significantly increased in liver, lungs and small intestine at various time points after CLP respectively (P < 0.05 or P < 0.01), while they didn't change in the kidneys. Treatment with AG490 could markedly inhibit HMGB1 mRNA expression in the liver and small intestine at 24 and 48 h (P < 0.05 or P < 0.01), and in lungs at 2 h following CLP (P < 0.01). Similarly, treatment with RPM significantly decreased HMGB1 mRNA expression in the lungs at 2, 6, 24 and 48 h, in the liver at 6 and 24 h, and in the small intestine at 24 and 48 h (P < 0.05 or P < 0.01). In addition, STAT1 and STAT3 activities in the liver and lungs were significantly correlated with corresponding tissue HMGB1 mRNA expression. CONCLUSIONS: Peritoneal infection could extensively activate STAT1 and limitedly activate STAT3 in vital organs. Activation of JAK/STAT pathway might be involved in up-regulating the gene expression of HMGB1 and systemic inflammation secondary to severe septic challenge.