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Anthocyanin generation in apples (Malus domestica) and the pigmentation that results from it may be caused by irradiation and through administration of methyl jasmonate (MeJA). However, their regulatory interrelationships associated with fruit coloration are not well defined. To determine whether MdERF109, a transcription factor (TF) involved in light-mediated coloration and anthocyanin biosynthesis, has synergistic effects with other proteins, we performed a yeast two-hybrid assessment and identified another TF, MdWER. MdWER was induced by MeJA treatment, and although overexpression of MdWER alone did not promote anthocyanin accumulation co-overexpression with MdERF109 resulted in significantly increase in anthocyanin biosynthesis. MdWER may form a protein complex with MdERF109 to promote anthocyanin accumulation by enhancing combinations between the proteins and their corresponding genes. In addition, MdWER, as a MeJA responsive protein, interacts with the anthocyanin repressor MdJAZ2. Transient co-expression in apple fruit and protein interaction assays allowed us to conclude that MdERF109 and MdJAZ2 interact with MdWER and take part in the production of anthocyanins upon MeJA treatment and irradiation. Our findings validate a role for the MdERF109-MdWER-MdJAZ2 module in anthocyanin biosynthesis and uncover a novel mechanism for how light and MeJA signals are coordinated anthocyanin biosynthesis in apple fruit.
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Acetatos , Antocianinas , Ciclopentanos , Frutas , Regulación de la Expresión Génica de las Plantas , Luz , Malus , Oxilipinas , Proteínas de Plantas , Ciclopentanos/metabolismo , Oxilipinas/metabolismo , Antocianinas/metabolismo , Antocianinas/biosíntesis , Acetatos/farmacología , Acetatos/metabolismo , Malus/metabolismo , Malus/genética , Malus/efectos de la radiación , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Frutas/metabolismo , Frutas/genética , Frutas/efectos de la radiación , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Reguladores del Crecimiento de las Plantas/metabolismoRESUMEN
An in planta chemical cross-linking-based quantitative interactomics (IPQCX-MS) workflow has been developed to investigate in vivo protein-protein interactions and alteration in protein structures in a model organism, Arabidopsis thaliana. A chemical cross-linker, azide-tag-modified disuccinimidyl pimelate (AMDSP), was directly applied onto Arabidopsis tissues. Peptides produced from protein fractions of CsCl density gradient centrifugation were dimethyl-labeled, from which the AMDSP cross-linked peptides were fractionated on chromatography, enriched, and analyzed by mass spectrometry. ECL2 and SQUA-D software were used to identify and quantitate these cross-linked peptides, respectively. These computer programs integrate peptide identification with quantitation and statistical evaluation. This workflow eventually identified 354 unique cross-linked peptides, including 61 and 293 inter- and intraprotein cross-linked peptides, respectively, demonstrating that it is able to in vivo identify hundreds of cross-linked peptides at an organismal level by overcoming the difficulties caused by multiple cellular structures and complex secondary metabolites of plants. Coimmunoprecipitation and super-resolution microscopy studies have confirmed the PHB3-PHB6 protein interaction found by IPQCX-MS. The quantitative interactomics also found hormone-induced structural changes of SBPase and other proteins. This mass-spectrometry-based interactomics will be useful in the study of in vivo protein-protein interaction networks in agricultural crops and plant-microbe interactions.
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Arabidopsis/metabolismo , Regulación de la Expresión Génica de las Plantas , Mapeo de Interacción de Proteínas/métodos , Proteoma/metabolismo , Proteínas Represoras/metabolismo , Secuencia de Aminoácidos , Arabidopsis/genética , Proteínas de Arabidopsis , Cromatografía Liquida , Reactivos de Enlaces Cruzados/química , Modelos Moleculares , Péptidos/análisis , Péptidos/química , Prohibitinas , Unión Proteica , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Estructura Secundaria de Proteína , Proteolisis , Proteoma/química , Proteoma/genética , Proteínas Represoras/química , Proteínas Represoras/genética , Coloración y Etiquetado/métodos , Succinimidas/química , Espectrometría de Masas en TándemRESUMEN
In order to study the status and sources of heavy metal pollution in Yinchuan Yellow River floodplain soils, we used inductively coupled plasma mass spectrometry (ICP-MS) to determine the presence of eight heavy metals in 92 soil samples from the Yinchuan Yellow River floodplain and used enrichment factors, geological accumulation index, and potential ecological risk index to analyze and evaluate the characteristics of heavy metal pollution in the study area. Combined correlation analysis, absolute factor analysis-multiple linear regression model (APCS-MLR), positive matrix factorization (PMF), and geostatistics were used to analyze the sources of soil heavy metals. The results showed that the content of eight heavy metals in the surface soil of the Yellow River floodplain in Yinchuan City were lower than the screening value of soil pollution risk in agricultural land; Cu and Pb contents were lower than the background value of Yinchuan City soil, and the contents of the remaining six elements were higher than the background value. The coefficients of variation of Zn and Cd were large and in the medium variation level and were influenced by anthropogenic activities. The heavy metal content varied between different land types and generally showed that wasteland > abandoned farmland > woodland > cultivated land. The average content of Cu and Pb in forest and arable soils was lower than the regional background value, whereas the rest of the heavy metals in different land types were higher than the soil background value. The analysis of enrichment factors showed that Zn and Cd were slightly enriched in the study area, and the cumulative index method and the evaluation of the potential risk of single heavy metals indicated that more than 60% of the sites in the study area were contaminated with Cd at a medium or higher potential ecological hazard. The comprehensive evaluation results of potential ecological risk showed that the overall ecological risk level of the study area was mild. From the distribution of heavy metal ecological risk comprehensive index sample points, only one point was in moderate ecological hazard, and the pollution point showed very few. Comprehensive correlation analysis, APCS-MLR model, PMF model, and geostatistical analysis results confirmed that Zn and Cd in the study area were mainly derived from human activities such as agricultural activities and transportation, and the remaining heavy metals were derived from soil parent materials. The results of this study can provide a scientific basis for the ecological protection and sustainable development of the Yellow River in Yinchuan City.
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BACKGROUND: The Toronto Adolescent and Youth (TAY) Cohort Study will characterize the neurobiological trajectories of psychosis spectrum symptoms, functioning, and suicidality (i.e., suicidal thoughts and behaviors) in youth seeking mental health care. Here, we present the neuroimaging and biosample component of the protocol. We also present feasibility and quality control metrics for the baseline sample collected thus far. METHODS: The current study includes youths (ages 11-24 years) who were referred to child and youth mental health services within a large tertiary care center in Toronto, Ontario, Canada, with target recruitment of 1500 participants. Participants were offered the opportunity to provide any or all of the following: 1) 1-hour magnetic resonance imaging (MRI) scan (electroencephalography if ineligible for or declined MRI), 2) blood sample for genomic and proteomic data (or saliva if blood collection was declined or not feasible) and urine sample, and 3) heart rate recording to assess respiratory sinus arrhythmia. RESULTS: Of the first 417 participants who consented to participate between May 4, 2021, and February 2, 2023, 412 agreed to participate in the imaging and biosample protocol. Of these, 334 completed imaging, 341 provided a biosample, 338 completed respiratory sinus arrhythmia, and 316 completed all 3. Following quality control, data usability was high (MRI: T1-weighted 99%, diffusion-weighted imaging 99%, arterial spin labeling 90%, resting-state functional MRI 95%, task functional MRI 90%; electroencephalography: 83%; respiratory sinus arrhythmia: 99%). CONCLUSIONS: The high consent rates, good completion rates, and high data usability reported here demonstrate the feasibility of collecting and using brain imaging and biosamples in a large clinical cohort of youths seeking mental health care.
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Proteómica , Trastornos Psicóticos , Niño , Humanos , Adolescente , Estudios de Cohortes , Neuroimagen , EncéfaloRESUMEN
Aquaporin-8 (AQP8), a member of the aquaporin family, is strongly expressed in follicular granulosa cells, which could affect the hormone secretion level in females. AQP8, as a membrane protein, could mediate H2O2 into cells, thereby triggering various biological events. The deficiency of Aqp8 increases female fertility, resulting from the decrease in follicular atresia. The low cell death rate is related to the apoptosis of granulosa cells. However, the mechanism by which AQP8 regulates the autophagy of granulosa cells remains unclear. Thus, this study aimed to explore the effect of AQP8 on autophagy in follicular atresia. We found that the expression of the autophagy marker light-chain protein 3 was significantly downregulated in the granulosa cells of Aqp8-knockout (Aqp8 -/- ) mice, compared with wild-type (Aqp8 +/+ ) mice. Immunofluorescence staining and transmission electron microscopic examination indicated that the number of autophagosomes in the granulosa cells of Aqp8 -/- mice decreased. Using a follicular granulosa cell autophagy model, namely a follicular atresia model, we verified that the concentration of H2O2 significantly increased during the autophagy of granulosa cells, consistent with the Aqp8 mRNA level. Intracellular H2O2 accumulation was modulated by endogenous AQP8 expression level, indicating that AQP8-mediated H2O2 was involved in the autophagy of granulosa cells. AQP8 deficiency impaired the elevation of H2O2 concentration through phosphorylated tyrosine activation. In addition, we carried out the analysis of transcriptome sequencing datasets in the ovary and found there were obvious differences in principal components, differentially expressed genes (DEGs) and KEGG pathways, which might be involved in AQP8-regulated follicular atresia. Taken together, these findings indicated that AQP8-mediated H2O2 transport could mediate the autophagy of granulosa cells. AQP8 might be a potential target for diseases related to ovarian insufficiency.
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Low temperature can affect the growth and development of plants through changes in DNA demethylation patterns. Another known effect of low temperature is the accumulation of anthocyanin pigments. However, it is not known whether the two phenomena are linked, specifically, whether DNA demethylation participates in anthocyanin accumulation in response to low-temperature stress. The ROS1 gene is involved in plant DNA demethylation and influences methylation levels in response to low temperature stress. In this study, using RNA sequencing, we detected that the transcription levels of MdROS1 correlate with the anthocyanin content, as well as with those of anthocyanin biosynthesis-related genes in apple (Malus domestica), at low temperatures. Genomic bisulfite sequencing showed that the methylation levels of the promoters of the anthocyanin related genes MdCHS, MdCHI, MdF3'H, MdANS, MdUFGT, and MdMYB10 decreased in apple leaves after low-temperature treatment. Similar expression and methylation results were also found in apple fruit. Transiently silencing MdROS1 in the leaves and fruit of apple cultivars inhibited the accumulation of anthocyanins and led to decreased expression of anthocyanin biosynthetic genes, and the opposite results were detected in MdROS1-overexpressing leaves and fruit. A promoter binding assay showed that the conserved RRD-DME domains of MdROS1 directly bind to the promoters of MdF3'H and MdUFGT. Taken together, these results suggest that ROS1 affects the anthocyanin biosynthetic pathway by decreasing the methylation level of anthocyanin-related gene promoters, thereby increasing their expression and increasing anthocyanin accumulation.
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Optimal performance in many tasks requires minimizing the impact of both visual distraction and mind-wandering. Yet, so far, these two types of distraction have been studied in isolation and it remains unclear whether they act in similar or dissociable ways across age groups. Here, we studied the impact of visual distraction and mind-wandering on performance in a go/no-go task in young and older adults. Older adults reported higher task focus than young, which was associated with a specific age-related reduction in mind-wandering, rather than to thoughts triggered by the task. Older adults exhibited fewer no-go errors, higher mean reaction time (RT) and reduced RT variability compared to young adults. In contrast, visual distraction was associated with a disproportionate effect in older versus young adults on go accuracy, mean RT, and RT variability. Decreasing task focus was similarly associated with reduced go- and no-go accuracy and increased RT variability across age groups. In summary, our results suggest that whereas older adults are disproportionately affected by visual distraction compared to young, they exhibit a reduction in mind-wandering frequency. Moreover, the impact of decreasing task focus on task performance is similar across age groups. Our results suggest a dissociation of the impact of visual distraction and mind-wandering as a function of age. (PsycInfo Database Record (c) 2020 APA, all rights reserved).
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Atención/fisiología , Inhibición Psicológica , Análisis y Desempeño de Tareas , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Envejecimiento , Femenino , Humanos , Masculino , Persona de Mediana Edad , Tiempo de Reacción , Adulto JovenRESUMEN
Optimal performance in many tasks requires minimizing the impact of both visual distractors in the environment and distracting internal thoughts (i.e., mind-wandering). Prior research has indicated that older adults are disproportionately affected by the presence of visual distractors compared to young adults, but are not excessively affected by distracting thoughts. Yet an explanation for these dissociable effects remains elusive. In the current study, we assessed age-related differences in event-related potentials and neural variability associated with internal distraction and visual distractors in a go/no-go task. Compared to young adults, older adults showed an increased visual distraction cost in mean reaction time (RT) and RT variability but a reduction in internal distraction frequency and a reduced internal distraction cost on go accuracy and RT variability. Visual distraction and internal distraction were associated with opposite patterns of behavioral and neural effects. Behaviorally, across age groups, internal distraction was associated with more no-go errors whereas visual distraction was associated with reduced no-go errors. Across groups, internal distraction was associated with decreased P3 amplitude, whereas visual distraction was associated with increased P3 amplitude. In addition, internal distraction was associated with an increase in neural variability (more so in young versus older adults), while visual distraction was associated with a reduction in variability in young adults only. We suggest that the opposing effects of the two distractor types on behavioral and neural measures occur because visual distraction is associated with increased attentional resources devoted to the task to overcome visual interference whereas internal distraction is associated with decreased attentional resources devoted to the task. Moreover, older adults exhibited reduced flexibility of neural variability as a function of both distractor types, which may correspond to a diminished ability to up-regulate attention in the face of visual distraction and a diminished shift in attention away from the task during internal distraction.
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Envejecimiento/fisiología , Atención/fisiología , Encéfalo/fisiología , Potenciales Evocados , Anciano , Cognición , Femenino , Humanos , Masculino , Tiempo de Reacción , Adulto JovenRESUMEN
Proanthocyanidins (PAs) are a class of flavonoid compounds in plants that play many important roles in pest and disease resistance and are beneficial components of the human diet. The crabapple (Malus) provides an excellent model to study PA biosynthesis and metabolism; therefore, to gain insights into the PA regulatory network in Malus plants, we performed RNA-seq profiling of fruits of the 'Flame' cultivar at five sequential developmental stages. KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis showed that differentially expressed genes (DEGs) related to the functional category 'plant hormone signal transduction' were significantly enriched during fruit development. Further analysis showed that ethylene signal transduction pathway genes or response genes, such as ERS (ethylene response sensor), EIN3 (ETHYLENE INSENSITIVE 3) and ERFs (ethylene response factors), may play an important role in the regulatory network of PA biosynthesis. Additionally, 12 DEGs, including 10 ERFs, 1 MYB, and 1 bHLH transcription factor, associated with PA biosynthesis were identified using WGCNA. The expression patterns of these genes correlated with PA accumulation trends and transcriptome data from qRT-PCR analysis. The expression of RAP2-4 (RELATED TO APETALA 2-4) and RAV1 (related to ABI3/VP1), which belong to the ERF transcription factor family, showed the greatest correlations with PAs accumulation among the 12 identified TFs. Agrobacterium mediated-transient overexpression of the RAP2-4 led to an increase in PA abundance in crabapple leaves and apple fruits, and the opposite results were observed in RAV1-overexpressed crabapple leaves and apple fruits. Moreover, a yeast one-hybrid assay showed that RAP2-4 and RAV1 specifically bound the promoters of the PA biosynthetic genes McLAR1 and McANR2, respectively. These results indicate that RAP2-4 act as an inducer and RAV1 act as a repressor of PA biosynthesis by regulating the expression of the PA biosynthetic genes McLAR1 and McANR2. Taken together, we identified two potential regulators of PA biosynthesis and provide new insights into the ethylene-PA regulatory network.
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The deposition and dissolution of calcium carbonate can be affected by the action of biological factors, such as microbial-induced carbonate precipitation (MICP). Bacillus spp. has been isolated and applied to prevent soil erosion, increase the stability of slopes, dikes and dunes. However, previous studies have been always limited to a single calcium source (CaCl2) to evaluate the roles of bacteria, and the deposition and curing effect has not yet been quantified. Here, we designed deposition experiments to determine the effect of Bacillus cereus with different calcium sources and applied it to sand curing to measure the amount of deposition and curing. The results demonstrated that vaterite was produced when the Bacillus cereus participated. Also, more deposition was produced in the Ca(CH3COO)2 and CaCl2 groups, but the Ca(NO3)2 group showed optimal curing effects in the sand curing test due to the denser and more uniform deposition. This research will provide an important reference for the design and application of microbial-induced carbonate precipitation.
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Microorganism cells and spores are the main components of PM2.5 (fine particulate matter) as well as fine mineral particles. In the microscopic system, the microorganisms will affect the minerals through attachment, charge neutralization, and dissolution related to the cell surface structure and metabolite. To explore the process and the results of microbial cells and their extracellular polymeric substances (EPS) acting on the surface properties of minerals of PM2.5 through the metabolism, a common native soil bacterium Bacillus mucitaginosus with abundant extracellular polymers was chosen as the tested strain. Meanwhile, as one of the PM2.5 common minerals, calcite fine particles were taken as the research object to explore the influence of microbial cells and extracellular polymers on its surface properties. High performance liquid chromatography (HPLC), inductively coupled plasma spectrometry (ICP), Zeta potential analysis, Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction spectroscopy (XRD), and scanning electron microscopy (SEM) were used to characterize the composition of EPS, the soluble ions, surface charge, surface groups, crystal form, and surface morphology of calcite residual solid after being treated by the bacterial cells and EPS. The results revealed the EPS of B. mucitaginosus mainly consisted of protein and polysaccharides. Both the whole cell and its EPS could promote the dissolution of calcite particles into calcium ions. Due to the adhesion of organic groups on the calcite surface, the surface potential shifted significantly in the negative direction and the solution pH was clearly increased. The morphology of calcite surface was significantly changed after dissolution and re-crystallization. Experimental results also showed that the existence of the bacteria cells and EPS significantly affected the surface properties of calcite and provide a theoretical basis for the mechanism of PM fine particulate matter on human health impact for further study.
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Bacillus/química , Carbonato de Calcio/química , Material Particulado/química , Contaminantes Atmosféricos/química , Bacillus/fisiología , Cristalización , Concentración de Iones de Hidrógeno , Microscopía Electrónica de Rastreo , Polímeros/química , Polisacáridos/química , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de Superficie , Difracción de Rayos XRESUMEN
In this research, a zinc oxide micro-/nano-structured hollow sphere (MNHS) with a large specific surface area is applied as energy storage material to encapsulate poly(vinyl chloride) solution and control the fuel release. The sustained release effect of MNHS not only makes the motion more controllable, but enhances the motion time and distance.
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Driven by the Marangoni effect, a poly(vinyl chloride) (PVC) particle runs in its orbit (a) with high velocity due to the release of surfactant and heat. The PVC particles are also able to efficiently drive an aluminum bulk and to induce spinning and quick runs on a water surface (b).
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Norcantharidin (NCTD), the demethylated analog of the Chinese medicine cantharidin, exhibits anti-myeloma activity by inactivating nuclear factor-κB (NF-κB), which is implicated in multiple myeloma (MM) cell survival and resistance to bortezomib (BTZ). We investigated whether NCTD could potentiate the anti-tumor activity of BTZ in MM. NCTD inhibited the proliferation of MM cells and potentiated the anti-myeloma effects of BTZ by down-regulating IKKα and p-IκBα, which induced the accumulation of IκBα and inhibited the constitutive activation of NF-κB. This effect was correlated with the suppression of NF-κB-regulated gene products. Furthermore, a chemotherapy-potentiating effect of NCTD on BTZ was also observed in vivo. Our study demonstrated that NCTD and BTZ exhibit significant therapeutic effects on MM through the NF-κB signal pathway in vitro and in vivo. Future studies will investigate the combined effects of NCTD and BTZ in patients with MM.
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Ácidos Borónicos/farmacología , Compuestos Bicíclicos Heterocíclicos con Puentes/farmacología , Mieloma Múltiple/tratamiento farmacológico , Pirazinas/farmacología , Ensayos Antitumor por Modelo de Xenoinjerto , Animales , Antineoplásicos/farmacología , Apoptosis/efectos de los fármacos , Western Blotting , Bortezomib , Caspasas/metabolismo , Puntos de Control del Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Sinergismo Farmacológico , Citometría de Flujo , Humanos , Quinasa I-kappa B/metabolismo , Proteínas I-kappa B/metabolismo , Masculino , Ratones , Ratones Desnudos , Mieloma Múltiple/metabolismo , Mieloma Múltiple/patología , Inhibidor NF-kappaB alfa , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
OBJECTIVE: To explore the synergetic effect of norcantharidin (NCTD) and adriamycin (ADR) on the proliferation and apoptosis of multiple myeloma (MM) cells. METHODS: Human MM cell line U266 cells were treated with NCTD alone (10 µmol/L) or in combination with ADR (0.25 µmol/L). MTT and Annexin V/PI staining were used to determine cell viability and apoptosis. The protein expression of nuclear factor-κB P65 (NF-κB P65), phosphorylated NF-κB p65 (p-NF-κB p65), NF-κB P65 inhibitor IκBα, phosphorylated IκBα (p-IκBα), survivin, Bcl-2 and Bax were determined by Western blot. Immunohistochemistry was used to determine the expression of vascular endothelial growth factor (VEGF). RESULTS: (1) NCTD potentiated the cytotoxicity and pro-apoptotic effects induced by ADR. The combination of NCTD and ADR had synergistic anti-proliferation effect. (2) Combination of ADR and NCTD downregulated the expression of nuclear NF-κB P65 and cytoplasm p-IκBα induced by ADR. The expression of nuclear NF-κB P65 and cytoplasm p-IκBα decreased from 2.08 ± 0.29 and 0.39 ± 0.07 to 0.48 ± 0.08 and 0.02 ± 0.01 respectively, while the expression of the cytoplasm NF-κB P65 and IκBα were unchanged in the ADR alone group and the combined group. (3) The expression of survivin and bcl-2 decreased from 0.31 ± 0.05 and 0.23 ± 0.05 to 0.03 ± 0.02 and 0.05 ± 0.02, while the expression of Bax increased from 0.46 ± 0.06 to 0.62 ± 0.08 respectively in ADR alone group and combined group. (4) The positive rate of VEGF in ADR group and combination group were (44.6 ± 4.4)% and (27.0 ± 2.1)% respectively, indicating that NCTD could potentiate the inhibition effect on VEGF induced by ADR. CONCLUSIONS: The results suggest that NCTD can potentialize the chemosensitivity of multiple myeloma cells to ADR through regulating NF-κB/IκBα signaling pathway and NF-κB-regulated gene products including survivin, Bcl-2, Bax and VEGF.