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Mesenchymal stem cells have shown noticeable potential for unlimited self-renewal. They can differentiate into specific somatic cells, integrate into target tissues via cell-cell contact, paracrine effects, exosomes, and other processes and then regulate the target cells and tissues. Studies have demonstrated that transplantation of MSCs could decrease the expression and concentration of collagen in the liver, thereby reducing liver fibrosis. A growing body of evidence indicates that apoptotic MSCs could inhibit harmful immune responses and reduce inflammatory responses more effectively than viable MSCs. Accumulating evidence suggests that mitochondrial transfer from MSCs is a novel strategy for the regeneration of various damaged cells via the rescue of their respiratory activities. This study is aimed at reviewing the functions of MSCs and the related roles of the programmed cell death of MSCs, including autophagy, apoptosis, pyroptosis, and ferroptosis, as well as the regulatory pathogenic mechanisms of MSCs in liver fibrosis. Research has demonstrated that the miR-200B-3p gene is differentially expressed gene between LF and normal liver samples, and that the miR-200B-3p gene expression is positively correlated with the degree of liver fibrosis, suggesting that MSCs could inhibit liver fibrosis through pyroptosis. It was confirmed that circulating monocytes could deliver MSC-derived immunomodulatory molecules to different sites by phagocytosis of apoptotic MSCs, thereby achieving systemic immunosuppression. Accordingly, it was suggested that characterization of the programmed cell death-mediated immunomodulatory signaling pathways in MSCs should be a focus of research.

High-dimensional self-exciting point processes have been widely used in many application areas to model discrete event data in which past and current events affect the likelihood of future events. In this paper, we are concerned with detecting abrupt changes of the coefficient matrices in discrete-time high-dimensional self-exciting Poisson processes, which have yet to be studied in the existing literature due to both theoretical and computational challenges rooted in the non-stationary and high-dimensional nature of the underlying process. We propose a penalized dynamic programming approach which is supported by a theoretical rate analysis and numerical evidence.

Ya-Chun Chu,1,2,* Hsu-Wei Fang,2,3,* Yu-Yi Wu,1 Yu-Jun Tang,1 Erh-Hsuan Hsieh,1 YiZhou She,4 Che-Yi Chang,1 I-Chan Lin,5,6 Yin-Ju Chen,1,7– 9 Guei-Sheung Liu,1,10– 12 Ching-Li Tseng1,8,9,13 1Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei City, Taiwan; 2Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei City, Taiwan; 3Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Miaoli County, Taiwan; 4School of Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei City, Taiwan; 5Department of Ophthalmology, Wan Fang Hospital, Taipei Medical University, Taipei City, Taiwan; 6Department of Ophthalmology, School of Medicine, College of Medicine, Taipei Medical University, Taipei City, Taiwan; 7Department of Radiation Oncology, Taipei Medical University Hospital, Taipei City, Taiwan; 8International Ph.D. Program in Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei City, Taiwan; 9Center for Precision Medicine and Translational Cancer Research, Taipei Medical University Hospital, Taipei City, Taiwan; 10Centre for Eye Research Australia, Royal Victorian Eye and Ear Hospital, East Melbourne, VIC, Australia; 11Ophthalmology, Department of Surgery, University of Melbourne, East Melbourne, VIC, Australia; 12Menzies Institute for Medical Research, University of Tasmania, Hobart, TAS, Australia; 13Research Center of Biomedical Device, College of Biomedical Engineering, Taipei Medical University, Taipei City, Taiwan*These authors contributed equally to this workCorrespondence: Ching-Li Tseng, Tel +886 2 2736 1661 (ext. 5214), Email chingli@tmu.edu.twBackground: Corneal neovascularization (NV) is a process of abnormal vessel growth into the transparent cornea from the limbus and can disturb the light passing through the cornea, resulting in vision loss or even blindness. The use of nanomedicine as an effective therapeutic formulation in ophthalmology has led to higher drug bioavailability and a slow drug release rate. In this research, we designed and explored the feasibility of a new nanomedicine, gp91 ds-tat (gp91) peptide-encapsulated gelatin nanoparticles (GNP-gp91), for inhibiting corneal angiogenesis.Methods: GNP-gp91 were prepared by a two-step desolvation method. The characterization and cytocompatibility of GNP-gp91 were analyzed. The inhibition effect of GNP-gp91 on HUVEC cell migration and tube formation was observed by an inverted microscope. The drug retention test in mouse cornea was observed by in vivo imaging system, fluorescence microscope, and DAPI/TAMRA staining. Finally, the therapeutic efficacy and evaluation of neovascularization-related factors were conducted through the in vivo corneal NV mice model via topical delivery.Results: The prepared GNP-gp91 had a nano-scale diameter (550.6 nm) with positive charge (21.7 mV) slow-release behavior (25%, 240hr). In vitro test revealed that GNP-gp91 enhanced the inhibition of cell migration and tube formation capacity via higher internalization of HUVEC. Topical administration (eyedrops) of the GNP-gp91 significantly prolongs the retention time (46%, 20 min) in the mouse cornea. In chemically burned corneal neovascularization models, corneal vessel area with a significant reduction in GNP-gp91 group (7.89%) was revealed when compared with PBS (33.99%) and gp91 (19.67%) treated groups via every two days dosing. Moreover, GNP-gp91 significantly reduced the concentration of Nox2, VEGF and MMP9 in NV’s cornea.Conclusion: The nanomedicine, GNP-gp91, was successfully synthesized for ophthalmological application. These data suggest that GNP-gp91 contained eyedrops that not only have a longer retention time on the cornea but also can treat mice corneal NV effectively delivered in a low dosing frequency, GNP-gp91 eyedrops provides an alternative strategy for clinical ocular disease treatment in the culture.Keywords: gp91 peptide, gelatin, nanoparticles, ocular retention, control release, corneal neovascularization, eye drops

In recent years, with the development of ’China’s new urbanization, the “characteristic town movement” with the development of industrial economy first has brought problems to a large number of rural settlements, such as no cultural planning, no consumption of industry, and no soul. Then, in reality, there are still a large number of rural settlements under the planning of the upper-level local government, with the goal of developing into a characteristic town in the future. Therefore, this study believes that there is an urgent need to build a framework for evaluating the construction potential of rural settlements with sustainable characteristic towns. Not only that but also a decision analysis model should be provided for real-world empirical cases. This model needs to cover the assessment of the sustainable development potential of characteristic towns as the goal and the formulation of improvement strategies. This study combines the data collection of current characteristic town development rating reports, applies data exploration technology to extract core impact elements and obtain hierarchical decision rules, integrates expert domain knowledge with DEMATEL technology, and establishes an impact network relationship diagram between core impact elements. At the same time, the representative characteristic town cases are assessed for their sustainable development potential, and the modified VIKOR technique is applied to clarify the actual problems of the empirical cases, in an attempt to determine whether the development potential and development plan of the characteristic town meet the sustainable development needs from the pre-evaluation mechanism.

Ir3 is reported to photodegrade CAIX under hypoxia, which improve the cancer immune microenvironment, and induce immunogenic pyroptosis through lipid peroxidation under hypoxia.

Irregular mare patches (IMPs), recently discovered on the Moon, are eruptions of magma on the lunar surface, and their origins are still in question. While prior studies on IMPs have mainly focused on optical image analysis, in this study, an analysis of the characteristics of minerals is performed exemplary for the first time. Modified Gaussian model (MGM) deconvolution was applied to the infrared spectrum to confirm the properties of the mafic mineral. Mafic minerals were analyzed for 6 olivine-rich (Ol-rich) IMPs out of 91 currently reported, and only 4 of them yielded results of significance. All four sites showed more abundance of Fe than Mg, and manifested a weak relationship with Mg-suite rock. However, a problem was discovered during the MGM application process due to pilot implementation. In order to solve this problem, it is required to adjust the MGM initial condition settings more precisely and to increase the signal to noise ratio of the observation data. Moreover, it is necessary to analyze the mineral properties for all IMPs considering minerals other than Ol and utilize them to deduce the origin of the IMPs.

Ni-containing heteropolyvanadate, Na

In the setting of acetabular dysplasia, the increased translational motion of the femur may damage the labrum and cartilage, as well as stretch the capsule. The purpose of the study was to investigate the relationship between the acetabular coverage and the capsular stiffness by assessing the distension of anterior and posterior joint recesses on the hip computed tomography arthrography.One hundred thirty-three patients (138 hips) with a median age of 36 years (range 18-50 years) who received the computed tomography arthrography for evaluation of nonarthritic hip pain in our institute between 2015 and 2017 were retrospectively reviewed. The maximal distance between the anterior/posterior capsule and the anterior femoral head-neck junction/posterior femoral head on the axial imaging of computed tomography arthrography was defined as the width of anterior/posterior joint recess. The width of anterior/posterior joint recess was adjusted with the diameter of the femoral head and was then compared between acetabular dysplasia (lateral center-edge angle 25°), normal acetabulum (lateral center-edge angle between 25 and 39°), and deep acetabulum (lateral center-edge angle 39°). In addition, the standard univariate linear regression analysis was used to investigate the relationship between the adjusted width of anterior/posterior joint recess and anterior/posterior coverage of the hip, determined by the anterior/posterior wall index.The adjusted width of posterior joint recess was significantly greater in the acetabular dysplasia group than the normal acetabulum and deep acetabulum groups (p 0.01 and p = 0.02, respectively). There was no significant difference of the adjusted width of anterior joint recess between the groups (n.s.). The adjusted width of posterior joint recess had a significant but weak negative correlation with the anterior wall index (r = - 0.25, p 0.001), and no correlation with the posterior wall index (r = - 0.0004, n.s.). There was no significant correlation between the adjusted width of anterior joint recess and the anterior/posterior wall index (r = 0.05, n.s./r = 0.07, n.s.).The distension of posterior capsule on the computed tomography arthrography was significantly greater in acetabular dysplasia. In addition, there was a significant but weak negative correlation between the distension of posterior capsule and the anterior coverage of the hip. It indicated a looser posterior capsule was observed in a dysplastic hip. The relevance of posterior capsular laxity to clinical outcomes warrants further investigation. Given the fact that the distension of anterior capsule was not significantly higher in acetabular dysplasia, the need of anterior capsular plication in a dysplastic hip should be carefully evaluated.Level III.

Poly(ADP-ribose) polymerase (PARP) inhibitors have demonstrated promising clinical activity in multiple cancers. However, resistance to PARP inhibitors remains a substantial clinical challenge. In the present study, we report that anaplastic lymphoma kinase (ALK) directly phosphorylates CDK9 at tyrosine-19 to promote homologous recombination (HR) repair and PARP inhibitor resistance. Phospho-CDK9-Tyr19 increases its kinase activity and nuclear localization to stabilize positive transcriptional elongation factor b and activate polymerase II-dependent transcription of HR-repair genes. Conversely, ALK inhibition increases ubiquitination and degradation of CDK9 by Skp2, an E3 ligase. Notably, combination of US Food and Drug Administration-approved ALK and PARP inhibitors markedly reduce tumor growth and improve survival of mice in PARP inhibitor-/platinum-resistant tumor xenograft models. Using human tumor biospecimens, we further demonstrate that phosphorylated ALK (p-ALK) expression is associated with resistance to PARP inhibitors and positively correlated with p-Tyr19-CDK9 expression. Together, our findings support a biomarker-driven, combinatorial treatment strategy involving ALK and PARP inhibitors to induce synthetic lethality in PARP inhibitor-/platinum-resistant tumors with high p-ALK–p-Tyr19-CDK9 expression.

Background: Pediatric-onset multiple sclerosis (POMS) represents the earliest stage of disease pathogenesis. Investigating the cerebrospinal fluid (CSF) proteome in POMS may provide novel insights into early MS processes. Objective: To analyze CSF obtained from children at time of initial central nervous system (CNS) acquired demyelinating syndrome (ADS), to compare CSF proteome of those subsequently ascertained as having POMS versus monophasic acquired demyelinating syndrome (mADS). Methods: Patients were selected from two prospective pediatric ADS studies. Liquid chromatography–mass spectrometry (LC-MS) was performed in a Dutch discovery cohort (POMS n = 28; mADS n = 39). Parallel reaction monitoring–mass spectrometry (PRM-MS) was performed on selected proteins more abundant in POMS in a combined Dutch and Canadian validation cohort (POMS n = 48; mADS n = 106). Results: Discovery identified 5580 peptides belonging to 576 proteins; 58 proteins were differentially abundant with ⩾2 peptides between POMS and mADS, of which 28 more abundant in POMS. Fourteen had increased abundance in POMS with ⩾8 unique peptides. Five selected proteins were all confirmed within validation. Adjusted for age, 2 out of 5 proteins remained more abundant in POMS, that is, Carboxypeptidase E (CPE) and Semaphorin-7A (SEMA7A). Conclusion: This exploratory study identified several CSF proteins associated with POMS and not mADS, potentially reflecting neurodegeneration, compensatory neuroprotection, and humoral response in POMS. The proteins associated with POMS highly correlated with age at CSF sampling.

Nitrilases are promising biocatalysts to produce high-value-added carboxylic acids through hydrolysis of nitriles. However, since the enzymes always show low activity and sometimes with poor reaction specificity toward 2-chloronicotinonitrile (2-CN), very few robust nitrilases have been reported for efficient production of 2-chloronicotinic acid (2-CA) from 2-CN. Herein, a nitrilase from Paraburkholderia graminis (PgNIT) was engineered to improve its catalytic properties. We identified the beneficial residues via computational analysis and constructed the mutant library. The positive mutants were obtained and the activity of the "best" mutant F164G/I130L/N167Y/A55S/Q260C/T133I/R199Q toward 2-CN was increased from 0.14 × 10

The small-scale magnetospheric and ionospheric plasma experiment (SNIPE) is a mission initiated by the Korea Astronomy and Space Science Institute (KASI) in 2017 and comprises four 6U-sized nano-satellites (Korea Astronomy and Space Science Institute Satellite-1, KASISat-1) flying in formations. The main goal of the SNIPE mission is to investigate the space environment in low Earth orbit at 500-km. Because Iridium & GPS Board (IGB) is installed on the KASISat-1, a communication simulation is required to analyze the contact number and the duration. In this study, communication simulations between the Iridium satellite network and KASISat-1 are performed using STK Pro (System Tool Kit Pro Ver 11.2) from the AGI (Analytical Graphics, Inc.). The contact number and durations were analyzed by each orbit and date. The analysis shows that the average access number per day is 38.714 times, with an average of 2.533 times per orbit for a week. Furthermore, on average, the Iridium satellite communication is linked for 70.597 min daily. Moreover, 4.625 min is the average duration of an individual orbit.

We have developed an iron-catalyzed synthesis of pyrrolo[2,1-a]isoquinoline derivatives with tetrahydroisoquinolines, arylacyl bromides, and nitroolefins. Highly functionalized pyrrolo[2,1-a]isoquinolines can be obtained in moderate to good yields through a three-component N-alkylation/oxidative 1,3-dipolar cycloaddition/elimination/aromatization cascade. The obtained products in this study can be easily modified by easy chemical transformations to structurally complex molecules bearing privileged framework.

In the manufacturing of battery boxes using the aluminum extruded process, poor consistency of products and a short life of the die for making aluminum structural sections are usually observed. A new method of producing battery boxes is proposed that combines steel stamped and aluminum extruded process. This paper first describes the design requirements for a battery box using a new process, and several important issues such as weld seam arrangement and error proofing in the manufacturing process are discussed. To address the issue of weld seam arrangement, the following three principles should be considered in the design: These principles include that the profile lap angle should be above 90°, three or more beams should not be lapped too closely together, and multiple brackets in close proximity should be designed as one unit. Secondly, according to the maximum external dimensions of the box and the relative position of each structure, a new box was designed in two parts: the external frame and the internal frame. In the box design, the external frame is designed from both the external frame mating surface and the external mount reinforcement, while the internal frame is solely described from the design of the internal frame mating surface. Finally, the newly designed box is subjected to finite element simulation analysis, a manufacturing process study, test verification, and bead optimization. The results show that the performance of the new battery box meets the design requirements. The study of composite process battery boxes in this paper is of guiding significance for the design of battery boxes.

As a catadromous fish, Asian sea bass (Lates calcarifer) juveniles migrate from seawater (SW) to freshwater (FW) for growth and development. During migration, they undergo physiological changes to acclimate to environmental salinity. Thus, it is crucial to understand how SW-to-FW migration affects the gut microbiota of catadromous fish. To the best of our knowledge, no study has revealed the effects of transfer to hypotonic environments on a catadromous fish microbiota. In this study, we aimed to determine the effects of FW transfer on the microbiota and cytokine gene expression in the intestines of juvenile catadromous Asian sea bass. The relationship between the water and the gut microbiota of this euryhaline species was also examined. We found that FW transfer affected both mucosa- and digesta-associated microbiota of Asian sea bass. Plesiomonas and Cetobacterium were dominant in both the mucosa- and digesta-associated microbiota of FW-acclimated sea bass. The pathogenic genera Vibrio, Staphylococcus, and Acinetobacter were dominant in the SW group. Although dominant fish microbes were present in the water, fish had their own unique microbes. Vitamin B6 metabolism was highly expressed in the FW fish microbiota, whereas arginine, proline, and lipid metabolism were highly expressed in the SW fish microbiota. Additionally, the correlation between cytokine gene expression and microbiota was found to be affected by FW transfer. Taken together, our results demonstrated that FW transfer altered the composition and functions of mucosa- and digesta-associated microbiota of catadromous Asian sea bass intestines, which correlated with cytokine gene expression.

IntroductionXuanhuang Pill (XHP) is a traditional Chinese medicine oral formula composed of 10 herbs. This study aims to verify the hepatoprotective activity of XHP and explain its possible mechanism.MethodsThe hepatoprotective activity of XHP was evaluated by constructing a mouse model of alcoholic liver disease, and the mechanism of XHP was preliminarily explained by utilizing ultra-performance liquid chromatography/time-of-flight mass spectrometry (UPLC-QTOF/MS), proteomics and network pharmacology.ResultsThe current study demonstrated that treatment with XHP ameliorated acute alcohol-induced liver injury in mice by significantly reducing alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and triglycerides (TGs) and malondialdehyde (MDA) content. Remarkably, treatment also increased superoxide dismutase (SOD) activity and glutathione (GSH) content. UPLC-QTOF/MS, 199 compounds were identified as within the make-up of the XHP. Network pharmacology analysis showed that 103 targets regulated by 163 chemical components may play an important role in the protective liver effect mediated by XHP. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis suggest that the HIF-1, FoxO, PI3K-Akt, insulin, and thyroid hormone signaling pathways are key modulators of XHP’s effects. Finally, eight key targets including Mapk1, Mapk3, Akt1, Map2k1, Pik3ca, Pik3cg, Raf1, and Prkca were verified by molecular docking and proteomics analysis, which provide insight into the hepatoprotective effect observed with XHP treatment.ConclusionIn summary, these results improved upon knowledge of the chemical composition and the potential mechanisms of hepatoprotective action of oral XHP treatment, providing foundational support for this formulation as a viable therapeutic option for alcoholic liver disease.

Purpose: Suppressor T cells are one of the determinants of colorectal cancer (CRC) clinical outcome. LAP+CD4+ T cell is a recently identified subset of suppressor T cells. This study was designed to investigate their clinical relevance in patients with CRC.Experimental Design: Sixty patients with CRC and 24 healthy donors (HD) were enrolled in this study. The percentages of LAP+CD4+ T cells in peripheral blood and tumor tissue were measured. The phenotype and functional relevance of LAP+CD4+ T cells were analyzed subsequently.Results: The percentages of LAP+CD4+ T cells in peripheral blood of patients with CRC were significantly higher than HD (HD vs. CRC: 3.1% ± 0.78% vs. 8.8% ± 5.8%, P < 0.0001) and in tumor tissue when compared with nontumor tissue (nontumor vs. tumor: 3.2% ± 1.1% vs. 9.5% ± 5.5%, P = 0.0002). In addition, LAP+CD4+ T cells with effector memory (EM) phenotype were more likely to accumulate in the tumor sites than in peripheral blood. These LAP+CD4+ T cells produced significantly higher levels of IFN-γ, IL-17 and comparatively lower IL-2 and very few IL-10. LAP+CD4+ T cells could suppress the proliferation of LAP−CD4+ T cells that were partially mediated by TGF-β. Furthermore, these LAP+CD4+ T cells accumulated in tumor site and increased further in the peripheral blood in patients with metastasis.Conclusions: LAP+CD4+ T cells as a suppressor subset could accumulate in the tumor microenvironment and circulated more in the peripheral blood with tumor progression in patients with CRC. Clin Cancer Res; 18(19); 5224–33. ©2012 AACR.

Targeting immune checkpoints such as programmed cell death 1 (PD-1) and programmed cell death ligand 1 (PD-L1) has transformed cancer treatment, with durable clinical responses across a wide range of tumor types. However, a high percentage of patients fail to respond to anti–PD-1/PD-L1 treatment. A greater understanding of PD-L1 regulation is critical to improving the clinical response rate of PD-1/PD-L1 blockade. Here, we demonstrate that PD-L1 is phosphorylated and stabilized by casein kinase 2 (CK2) in cancer and dendritic cells (DC). Phosphorylation of PD-L1 at Thr285 and Thr290 by CK2 disrupted PD-L1 binding with speckle-type POZ protein, an adaptor protein of the cullin 3 (CUL3) ubiquitin E3 ligase complex, protecting PD-L1 from CUL3-mediated proteasomal degradation. Inhibition of CK2 decreased PD-L1 protein levels by promoting its degradation and resulted in the release of CD80 from DC to reactivate T-cell function. In a syngeneic mouse model, combined treatment with a CK2 inhibitor and an antibody against T-cell immunoglobulin mucin-3 (Tim-3) suppressed tumor growth and prolonged survival. These findings uncover a mechanism by which PD-L1 is regulated and suggest a potential antitumor treatment option to activate DC function by blocking the CK2–PD-L1 pathway and inhibiting Tim-3.Significance:This work identifies a role for CK2 in immunosuppression by phosphorylation and stabilization of PD-L1, identifying CK2 inhibition as an immunotherapeutic approach for treating cancer.

Correlations between expression status of pEGFR, pSTAT3, CEBPD, and ABCB1 in muscle invasive UCUB in our independent cohort containing 60 patients receiving post-operative chemotherapy

CEBPD expression had significant worse MeFS in UCUB patients. A and B, Kaplan-Meier plots discloses the predictive significances of CEBPD immunoexpression for metastasis-free survival in UCUB.

Attenuated pSTAT3, CEBPD, ABCB1 and ABCC2 mRNA levels are observed in CDDP-resistant NTUB1/P xenografted NOD/SCID mice. A and B, Following 3 weeks of drug treatment, all animals were sacrificed, and their tumors were collected for protein and mRNA analysis. Protein samples were employed for Western blot analysis by using specific antibodies. The following data present changes in pSTAT3, CEBPD, and ABCB1 levels in NTUB1/P-xenografted NOD/SCID mice treated with CDDP and gefitinib or with CDDP and S3I-201. qPCR assays were performed using the total RNA harvested from tumor samples. In this figure, *** denotes a significant difference (P < 0.001). B, evaluation of body weight of experimental mice used in Fig. 5. CTL: control.

Levels of CEBPD associates with STAT3 activity in gefitinib-treated CDDP-resistant NTUB1/P (NTP) cells. A, Western blot analysis was conducted with lysates from NTP cells treated with gefitinib following various time courses and blotted with specific antibodies. B, gefitinib and S3I-201 attenuate expression of phosphorylation of STAT3 (pY705) and CEBPD in J82 cells. Cells were treated with CDDP alone or in combination with gefitinib or S3I-201 for 24 h. Western blot was conducted with indicated antibodies, respectively.

Correlations between CEBPD expression and other important clinicopathological parameters in 79 muscle invasive UCUB receiving post-operative chemotherapy

Expression and activities of EGFR and STAT3 levels in UCUB cells. Western blot analysis was conducted with lysates from UCUB cells. TSGH8301 (TSG), TCCSUP (TCC), NTUB1 (NTU) and CDDP resistance NTUB1/P (NTP).

Overexpression of CEBPD upregulates ABCB1 and ABCC2 transcription in CDDP-resistant NTUB1/P (NTP) cells. A, RT-PCR assay was conducted with total RNA harvested from NTP cells transfected with pCDNA3/HA/CEBPD expression vectors. B, overexpression of CEBPD upregulates ABCB1 transcription containing known exon 1 (NM_000927.4) in CDDP-resistant NTP cells. RT-PCR assay was conducted with total RNA harvested from NTP cells transfected with pCDNA3/HA/CEBPD expression vectors. Upper panel shows location of forward primer (+159) in exon 1 and reverse primer (+348) in exon 2 of ABCB1 gene locus.

Gefitinib and S3I-201 attenuate ABCB1 transcripts of and enhance CDDP sensitivity of J82 cells. A, cells were treated with CDDP alone or in combination with gefitinib or S3I-201 for 24 h. A qPCR assay was conducted with specific primers for ABCB1 gene. ***, significant difference (P < 0.001). B, J82 cells were then transfected with ABCB1 reporter (-21~+917) (AB-I, as shown in Fig. 3D) and treated with CDDP alone or in combination with gefitinib or S3I-201. After 24 h. A Luciferase assay was conducted using lysates of transfectants. ***, significant difference (P < 0.001). C, gefitinib and S3I-201 attenuate ABCB1 pump activity and enhance CDDP sensitivity of J82 cells. J82 cells were treated with CDDP alone or CDDP combining gefitinib or S3I-201 for 24 h, MDR dye-loading solution was added to each well for 4 h, and fluorescence intensities were detected by ELISA reader. ***, significant difference (P < 0.001). D, J82 cells were treated with CDDP alone or CDDP combining gefitinib or S3I-201 for 24 h. Death of experimental cells was examined by PI staining. ***, significant difference (P < 0.001). E, CEBPD contributes to drug resistance of cisplatin (CDDP) in nasopharyngeal carcinoma HONE1/R (CDDP-resistant HONE) cells. HONE1/R cells were resistant to CDDP treatment. HONE1 and HONE1/R cells were treated with CDDP in dose-dependent manner. Cell viability was examined by CCK8 assay. *, significant difference (P < 0.05). **, significant difference (P < 0.01). ***, significant difference (P < 0.001). F, CDDP induces expression of CEBPD and ABCB1. HONE1/R cells were treated with CDDP for 24 h and then lysates of experimental cells were analyzed by Western blot. G, HONE1/R cells were treated with gefitinib, S3I-201 or CDDP or in combination as indicated for 24 h, MDR dye-loading solution was added to each well for 4 h and fluorescence intensities were detected by ELISA reader. ***, significant difference (P < 0.001). H, S3I-201 enhances sensitivity of CDDP in HONE1/R cells. HONE1/R cells were treated with CDDP (20 μM) alone or in combination with S3I-201 (100 μM) for 24 h and death of experimental cells was examined by PI staining. ***, significant difference (P < 0.001).