Bismuth, Nitrogen-Codoped Carbon Dots as a Dual-Read Optical Sensing Platform for Highly Sensitive, Ultrarapid, Ratiometric Detection of Doxorubicin.

Doxorubicin (DOX) is a potent anticancer drug, but it has side effects on normal tissues, particularly myocardial cells. Therefore, it is crucial to detect the DOX concentration in body fluids for effective clinical treatment. In this work, N,Bi-codoped CDs (Bi,N-CDs) were synthesized through a one-step hydrothermal method to carbonize the raw materials of 2,4-dinitroaniline and bismuth nitrate. The resulting Bi,N-CDs showed a reduced emission at 490 nm and an enhanced emission at 590 nm in the presence of DOX. The ratio of fluorescence (FL) intensity (F590/F490) was found to be a reliable indicator of DOX concentration, ranging from 0.05 to 30 µM and 40-200 µM, with detection limits (LOD) of 34 and 24 nM, respectively. A ratiometric fluorescence nanoprobe was established for highly selective and sensitive detection of DOX using a specific electrostatic interaction and inner filter effect between Bi,N-CDs and DOX. Meanwhile, Bi,N-CDs exhibited a distinct color change ranging from yellow to orange-red when exposed to DOX, allowing for a colorimetric method to measure DOX levels in the range of 0.05-30 µM, with a detection limit of 169 nM. The probe was triumphantly used to monitor DOX in actual samples via a dual-mode optical sensing strategy. This study contributes to the development of heteroatom-doped CDs and expands their potential applications for detecting biological samples.

Sequence and expression regulation of the BCL2L2 gene in pigs.

B cell lymphoma-2-like 2 (BCL2L2), an important regulator of apoptosis, plays vital roles in several physiological processes, as revealed by studies in humans and mice. However, reports on pig BCL2L2 are few, and the encoding gene has not been identified experimentally. This study was designed to clone the porcine BCL2L2 gene and its alternative splicing (AS) transcripts using molecular biological techniques and to analyze the regulatory mechanisms underlying transcription and translation. The BCL2L2 cDNA (V1) was 807 bp in length and encoded a polypeptide of 193 aa containing four BCL-2 homology domains. A total of nine AS transcripts were obtained, among which V2 and V3 differed from V1 in the 5' untranslated region (UTR). The core promoter was mapped to a range of -1102 to -759 bp (the first nucleotide of the start codon was designated as +1). There were several functional cis-elements, including one SP1 and two C/EBPα binding sites at around -759 bp. AS in the 5' UTR is involved in the regulation of gene expression, as revealed by dual-luciferase reporter and western blot analysis, and the secondary structure of the 5' UTRs may be the reason for the differential expression of V1-3. At the same time, an upstream open reading frame (ORF) existed in each of the three 5' UTRs, was found to repress the expression of the main ORF. Additionally, the roles of porcine BCL2L2 in cell proliferation and apoptosis were preliminarily analyzed. The results will contribute to further characterizing the role of BCL2L2.

Erratum: A2780 human ovarian cancer cells with acquired paclitaxel resistance display cancer stem cell properties.

[This corrects the article DOI: 10.3892/ol.2013.1568.].

A novel carbon-nanodots-based theranostic nano-drug delivery system for mitochondria-targeted imaging and glutathione-activated delivering camptothecin.

Chemotherapy is severely limited by continuously decreased therapeutic efficacy and uncontrolled side effects on normal tissue, which can be improved by constructing a nanoparticle-based drug delivery system (DDS). Nevertheless, no studies have reported on DDS-based on carbon-nanodots (CNDs), combining subcellular organelle-targeted imaging/drug delivery, high drug loading content, and glutathione (GSH)-sensitive drug release into one system. Herein, the as-fabricated CNDs can be covalently conjugated with a mitochondria-targeting ligand (triphenylphosphine, TPP), a smart GSH-responsive disulfide linker (S-S), and the anticancer drug (camptothecin, CPT) to initially prepare a theranostic nano-DDS (TPP-CNDs-S-CPT) with the drug loading efficiency of 64.6 wt%. Owing to excellent water dispersibility, superior fluorescence properties, satisfactory cell permeability, and favorable biocompatibility, TPP-CNDs-S-CPT was successfully used for intracellular mitochondrial-targeted imaging in vitro. High intracellular GSH concentrations in tumor cells caused the cleavage of S-S, resulting in concomitant activation and release of CPT, as well as significant fluorescence enhancement. In vivo, TPP-CNDs-S-CPT exhibited lower biological toxicity and even higher tumor-activatable performance than free CPT, as well as specific cancer therapy with few side effects. The mitochondria-targeted ability and the precise drug-release in tumor make TPP-CNDs-S-CPT a hopeful chemotherapy prodrug, providing significant theoretical basis and data support for in-depth understanding and exploration of chemotherapeutic DDS-based on CNDs.

Changes in Retinal Vessel Flow after Small Incision Lenticule Extraction.

Objective: In order to analyze changes in retinal vessel flow after small incision lenticule extraction (SMILE). Methods: A total of 32 patients (62 eyes) who underwent SMILE were enrolled in this prospective study. Optical parameters, including vessel density (VD), and perfusion density (PD) of foveal, parafoveal, and perifoveal regions, respectively, were measured before surgery and at 1 day, 1 week, 1 month, and 3 months postoperation. Preoperative parameters and surgical parameters were recorded. Results: Significant decreases in VD and PD on postoperative day 1 were detected in all quadrants, both in 3 mm and in 6 mm regions (P < 0.001). One month after surgery, VD returned to preoperative levels. None of the preoperative and surgical parameters were significantly correlated with the VD and PD fluctuations (all P > 0.05). Conclusion. VD may decrease significantly with regional disparity 1 day after SMILE while recovering at 1 month. Elevation of intraocular pressure due to suction may account for such changes.

Treatment of MGMT promoter unmethylated glioblastoma with PD-1 inhibitor combined with anti-angiogenesis and epidermal growth factor receptor tyrosine kinase inhibitor: a case report.

Glioblastoma (GBM) is the most common primary central nervous system (CNS) malignancy in adults and is associated with poor prognosis, especially even worse in those with unmethylated MGMT promoter. Currently, maximal safe resection combined with temozolomide (TMZ) concurrent chemoradiotherapy and TMZ adjuvant chemotherapy has been considered the standard treatment for newly diagnosed GBM. The efficacy of drugs other than TMZ is currently undefined. With increasing understanding of the biological characteristics of GBM, more and more studies are being conducted on drug targets, such as specific signaling pathways and microenvironment. Herein, we report the case of a GBM patient with unmethylated MGMT promoter who was intolerant to TMZ, and underwent treatment with the combination of carelizumab, anlotinib, and oxitinib during radiotherapy according to results of whole-exome sequencing (WES) and the patient's condition. The progression-free survival (PFS) and overall survival (OS) for this case were respectively nearly 11 and 18 months, significantly exceeding the historical data and the tolerance of the treatment for this case without sever adverse effects was favorable. Our case provides clinical evidence supporting the efficacy of the above three drugs and radiotherapy, which may translate into novel individualized treatment strategies for GBM patients who are intolerant to TMZ.

Tumor treating fields combined with a poly (adenosine diphosphate-ribose) polymerase inhibitor during radiotherapy for rapidly progressing IDH-wildtype diffuse astrocytoma: a case report.

A 57-year-old woman was diagnosed with IDH-wildtype (IDHwt) astrocytoma (World Health Organization grade II) with the molecular characteristics of glioblastoma. She underwent concurrent radiotherapy and chemotherapy according to the Stupp protocol in combination with a multi-target antiangiogenic drug and additional intrathecal chemotherapy using methotrexate. During treatment, the patient's tumor showed rapid progression. The chemotherapy with temozolomide was stopped and replaced with radiotherapy combined with tumor treating fields (TTF), the poly (adenosine diphosphate-ribose) polymerase (PARP) inhibitor niraparib, and anlotinib. After the radiotherapy was completed, the symptoms of increased intracranial pressure and epilepsy were well controlled. Considering the patient's tolerance to the treatment, the combined therapy of TTF and anlotinib was continued, and osimertinib, a third-generation epidermal growth factor receptor-tyrosine kinase inhibitor with good permeability of the blood-brain barrier, was added. The patient was regularly followed up and had no obvious adverse drug reactions. Head magnetic resonance imaging (plain scan + enhanced scan) suggested that the lesions were stable. For rapidly progressing glioblastomas or histological grade II/III IDHwt astrocytomas, the combination of TTF and a PARP inhibitor during radiotherapy may have a synergistic effect on tumor control and is well tolerated by patients.

Facile synthesis of ratiometric fluorescent carbon dots for pH visual sensing and cellular imaging.

A novel ratiometric emission N, S dual-doped carbon dots (N, S-CDs) were facilely developed for pH visual sensing via one-step hydrothermal method. The proposed N, S-CDs displayed intrinsic pH-sensitive behavior and exhibited ratiometric fluorescence emission (F563 nm/F645 nm) characteristic with the variation of pH values. Interestingly, a significant red shift of emission wavelength can be observed at 645 nm along with the emission at 563 nm decreased accordingly when the pH changed from 3.0 to 1.0. Simultaneously, the fluorescence of N, S-CDs aqueous solution was visually varied from yellow to red. The ratiometric pH linear response was in the region of 3.6 to 2.4 and the pKa was 2.90. Moreover, the N, S-CDs hold unique optical properties, good reversibility, superior biocompatibility and low cytotoxicity, which was further employed to monitor the intracellular pH fluctuations through the visible fluorescence changes between yellow and red. All these findings demonstrated that N, S-CDs can be utilized as the visual biosensor platform for tracking pH variations in extremely acidic environments such as gastric juice, which provided novel insights for clinical medical disease diagnosis (e.g., stomach disease detection) and other biomedical fields.

Apatinib treatment for symptomatic pseudoprogression after standard treatment for glioblastoma multiforme: a case report.

Apatinib, a novel tyrosine kinase inhibitor, has anti-angiogenetic effect just as bevacizumab. Although bevacizumab has been used successfully in treating cerebral radiation necrosis, there has yet not any report on that apatinib can treat pseudoprogression with symptoms. Here we report a case of glioblastoma multiforme (GBM) patient with pseudoprogression after receiving the concurrent chemoradiotherapy, which was successfully treated by apatinib. A 51-year-old woman had multiple intracranial lesions (left parietal and right frontal), the primary left parietal lesion was surgically removed and was pathologically confirmed as glioblastoma (WHO grade IV). Then the patient received postoperative temozolomide with concurrent chemoradiotherapy. Three weeks after the radiotherapy, the patient experienced increased intracranial pressure and seizure. Magnetic resonance imaging (MRI) T1 enhancement examination showed an increase of abnormal enhancement range in the area of irradiation. After multiple disciplinary team (MDT) discussion, the patient was diagnosed with pseudoprogression after radiotherapy. Then she was given apatinib for 8 weeks at a dose of 500 mg qd. During the treatment period, the clinical symptoms and corresponding nerve images of the patient have been rapidly improved. In 12 months after the radiotherapy, progression of tumor in the primary site has not been discovered. Apatinib showed a good therapeutic effect and tolerance for the development of pseudoprogression advances with obvious symptoms.

Dual-ligand functionalized carbon nanodots as green fluorescent nanosensors for cellular dual receptor-mediated targeted imaging.

The conjugation of ligands to nanoparticles as drug delivery systems that target specific cells is a promising approach for the delivery of therapeutic agents to tumor cells. Herein, we prepared green-emission fluorescent carbon nanodots (CNDs) by a facile hydrothermal method with d-(+)-glucosamine hydrochloride and l-aspartic acid as the precursors, then covalently conjugated with folate (FA), polyethyleneimine (PEI) and hyaluronic acid (HA) to develop dual ligand-decorated nanocarriers (FA-PEI-HA-CNDs) for the targeted imaging of cancer cells. FA-PEI-HA-CNDs integrated the excellent fluorescence property of CNDs, and can be used for the real-time and noninvasive location tracking of cancer cells. The cellular uptake study demonstrated that FA-PEI-HA-CNDs markedly improved the internalization efficiency in A-549 cells via folate/CD44 receptor-mediated endocytosis in comparison with that of the A549 cells pretreated with excess FA, HA, and FA and HA. Therefore, these dual folate/CD44 receptor-targeted CNDs (FA-PEI-HA-CNDs) show promising potential for cancer detection, drug delivery, and individualized treatment as performance platforms.

A label-free nano-probe for sequential and quantitative determination of Cr(VI) and ascorbic acid in real samples based on S and N dual-doped carbon dots.

A fluorescent sulfur and nitrogen dual-doped carbon dots (S,N-CDs) was prepared by a simple and one-step acid-base neutralization and exothermic carbonization method. Hexavalent chromium (Cr(VI)) could effectively quench the fluorescence of S,N-CDs based on inner filter effect (IFE) and dynamic quenching, whereas ascorbic acid (AA) could recover the fluorescence of S,N-CDs/Cr(VI) because of IFE weakening. So an "on-off-on" and label-free nano-probe consecutive determination of Cr(VI) and AA was constructed. This nano-probe system demonstrated excellent selectivity and sensitivity to Cr(VI) and AA with linear range of 0.065-198 µmol/L (3.38-10,296 µg/L) and 6.6-892 µmol/L (1.16-157 mg/L), respectively. Meanwhile, the as-prepared S,N-CDs possess low toxicity and could be used for multi-color cell imaging in SMMC 7721 cells. More importantly, this nano-probe was successfully employed for detection of Cr(VI) in tap water and AA in food samples. In view of its simple detection condition, rapid response, wide linear range, low detection limit and inexpensive instrument, the as-constructed nano-probe system could have a wide range of potential application, including water quality monitoring and evaluation, food inspection and testing and biomedical analysis.

Sulforaphane-Conjugated Carbon Dots: A Versatile Nanosystem for Targeted Imaging and Inhibition of EGFR-Overexpressing Cancer Cells.

Because of the demand for precision medicine, the investigatation on the application of carbon-dots-based nanosystems in the field of biomedicine is attracting more and more attention. Therefore, we have built a multifunctional nanosystem based on sulforaphane-conjugated carbon dots (SFN-CDs) with thiourea skeleton and applied for EGFR-overexpressing cancer cells targeted imaging and inhibiting. The SFN-CDs are formed by grafting sulforaphane on the amino-rich yellow fluorescent carbon dots, which have excellent optical stability and can be distinguished from normal cells for targeted imaging of cancer cells. The vitro toxicity experiments demonstrated that the SFN-CDs can effectively inhibit EGFR-overexpressing cancer cell proliferation at concentrations below 100 µg mL-1. All these results validated that SFN-CDs nanoparticles with integration of diagnostic and therapeutic functions can be used as a potential nanodurg in early stage of cancer control. Moreover, this work provides useful insight into targeted nanoparticle design in the biological nanomedicine field.

Chemotherapy enhances tumor vascularization via Notch signaling-mediated formation of tumor-derived endothelium in breast cancer.

It is believed that tumor cells can give rise to endothelial cells and tumor endothelium has a neoplastic origin. Yet, the stimuli and underlying mechanism remain unclear. Here, we demonstrate that adriamycin or paclitaxel, first-line chemotherapy agent, induced breast cancer cells to generate morphological, phenotypical and functional features of endothelial cells in vitro. In xenografts models, challenges from adriamycin or paclitaxel induced cancer cells to generate the majority of microvessels. Importantly, in breast cancer specimens from patients with neoadjuvant anthracycline-based or taxane-based chemotherapy, tumor-derived endothelial microvessels, lined by EGFR-amplified or/and TP53+-CD31+ endothelial cells, was significantly higher in patients with progressive or stable disease (PD/SD) than in those with a partial or complete response (PR/CR). Further, exposure to the Notch signaling inhibitor and gene silencing studies showed that Notch signaling inhibition or silencing Nothc4/Dll3 decreased endothelial markers and function of tumor-derived endothelial cells under chemotherapy treatment, which may be through VEGFR3. Thus, our findings demonstrate that chemotherapy induces functional tumor-derived endothelial microvessels by mediating Notch signaling and VEGF signaling, and may provide new targets for anti-angiogenesis therapy in breast cancer.

MGMT promoter methylation in serum and cerebrospinal fluid as a tumor-specific biomarker of glioma.

O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation is a conventional technique to predict the prognosis or individualized treatment of glioma in tumor tissue following surgery or biopsy. However, the technique cannot be applied in those glioma patients with concomitant neurological dysfunctions or advanced age. The present study aimed to find a new minimally invasive and efficient alternative method for the detection of MGMT promoter methylation. The expression of MGMT promoter methylation was assessed in peripheral blood and cerebrospinal fluid (CSF), and compared to the corresponding tumor tissue from glioma patients. The 89 patients in the study [32 World Health Organization (WHO) grade II, 19 WHO grade III and 38 WHO grade IV) were pathologically-diagnosed glioma and received radiation therapy following sample collection. The resected glioma tumor tissue (89), corresponding serum (89) and CSF (78) samples were collected for the detection of MGMT promoter methylation using methylation-specific polymerase chain reaction. The sensitivity and specificity of detecting MGMT promoter methylation in CSF and serum were compared. Among the tumor tissue samples, 51/89 (57.3%) showed MGMT promoter methylation. The specificity of the detection in the CSF and serum samples reached 100%. The sensitivity of MGMT promoter methylation detection in CSF and serum were 26/40 (65.0%) and 19/51 (37.3%), respectively (P<0.05). In the WHO II, III and IV subgroups, the sensitivities of MGMT promoter methylation detection using CSF were 8/12 (66.7%), 11/18 (61.1%) and 7/10 (70.0%), respectively, which were significantly higher than the sensitivities using serum (7/21, 33.3%; 7/19, 36.8%; and 5/11, 45.5%, respectively P<0.05). Among patients with residual postoperative tumors, the sensitivities of detecting MGMT promoter methylation using CSF and serum were 18/25 (72.0%) and 10/24 (41.7%), respectively, both of which were significantly higher than the corresponding values for patients without residual tumors (8/15, 53.3% and 6/19, 31.6%, respectively; P<0.05). The detection of MGMT promoter methylation in CSF specimens shows higher sensitivity compared to the serum for glioma patients. Assessment of MGMT promoter methylation in CSF may provide a promising clinical methodology for early diagnosis, individual treatment, monitoring of recurrence and prognosis for glioma patients.

Acquisition of 5-fluorouracil resistance induces epithelial-mesenchymal transitions through the Hedgehog signaling pathway in HCT-8 colon cancer cells.

Colon cancer has a high incidence in individuals >60-years-old. The commonly used chemotherapeutic agent, 5-fluorouracil (5-FU), has gradually lost its potency in treating colorectal cancer following the acquisition of resistance. Drug resistance is usually associated with epithelial-mesenchymal transitions (EMTs) in cancer cells. In the present study, the EMT phenotypes of two colon cancer cell lines, wild-type (HCT-8/WT) and 5-FU-resistant (HCT-8/5-FU), were characterized following the analysis of cellular migration, proliferation, morphology and molecular changes. In order to further clarify the mechanism of EMT in HCT-8/5-FU cells, the effect of EMT pathway inhibitors upon drug sensitivity was investigated. The results revealed that the Hedgehog signaling pathway inhibitor, GDC0449, reversed drug resistance. Therefore, inhibition of the Hedgehog pathway may provide a novel chemotherapeutic strategy for the treatment of patients with 5-FU-resistant colon cancer.

A2780 human ovarian cancer cells with acquired paclitaxel resistance display cancer stem cell properties.

The use of chemotherapy to treat cancer is effective, but chemoresistance reduces this efficacy. Chemotherapy resistance involves several mechanisms, including the cancer stem cell (CSC) concept. The aim of the present study was to assess whether paclitaxel-resistant epithelial ovarian carcinoma is capable of generating cells with CSC-like properties. Using the paclitaxel-resistant A2780/PTX cell line, it was demonstrated that high aldehyde dehydrogenase 1 (ALDH1) activity identifies CSCs from diverse sources. Furthermore, the A2780/PTX cells had a strong ability to form colonies in soft agar assays. Notably, it was demonstrated that the inhibition of the PI3K signaling pathway abolished colony formation. These data suggest that there is a link between paclitaxel resistance and CSC enrichment. It is possible that therapeutic benefits, such as the restoration of chemosensitivity or the suppression of tumorigenicity, may be enabled by gaining further insights into the mechanisms underlying chemoresistance and the generation of CSCs.

Acquisition of paclitaxel resistance via PI3K­dependent epithelial­mesenchymal transition in A2780 human ovarian cancer cells.

Epithelial ovarian cancer is a major cause of mortality among women with gynecological malignancies. Paclitaxel is commonly used for chemotherapy of ovarian cancer, yet its efficacy is limited by chemoresistance. Generally, drug resistance is associated with acquisition of the epithelial-mesenchymal transition (EMT) in cancer. The aim of the present study was to determine whether the EMT is involved in acquired resistance to paclitaxel in A2780 human ovarian cancer cells. Using the paclitaxel-resistant A2780/PTX cell line, we examined the cellular morphology, molecular changes, migration and proliferation consistent with the EMT. Furthermore, we found that inhibition of phosphatidylinositol 3-kinase (PI3K) activity reduced the proliferation and migration and restored their sensitivity to paclitaxel. Our study provides new insights into EMT-like phenotypic changes that are linked to paclitaxel resistance in A2780 cells. We believe that inhibition of the PI3K signaling pathway could provide a novel therapeutic approach to overcome chemoresistance and prevent metastasis during paclitaxel chemotherapy.

Enhanced reactive oxygen species (ROS) yields and antibacterial activity of spongy ZnO/ZnFe2O4 hybrid micro-hexahedra selectively synthesized through a versatile glucose-engineered co-precipitation/annealing process.

In this study, sponge-like ZnO/ZnFe2O4 hybrid micro-hexahedra with diverse textures and compositions were fabricated by the thermal decomposition of hexahedral zinc/iron oxalate precursors, starting from a glucose-engineered co-precipitation process. The resulting ZnO/ZnFe2O4 micro-hexahedra were systematically characterized by X-ray powder diffraction, Fourier-transform infrared spectroscopy, scanning electronic microscopy, transmission electron microscopy (TEM), high-resolution TEM, and surface area analysis. Moreover, modulation in crystal size, composition, and textural properties of spongy ZnO/ZnFe2O4 micro-hexahedra was easily achieved by varying the Zn2+/Fe3+ feeding ratio and the annealing temperature. The antibacterial property of the products was analyzed by testing ATP (adenosine triphosphate) and inhibition zones. Results showed that oxidative stress was the governing mechanism for the antibacterial activity of ZnO/ZnFe2O4 hybrid materials. Moreover, we found that the higher reactive oxygen species yields and the resulting antibacterial activity were exhibited by the ZnO/ZnFe2O4 micro-hexahedra formed at lower sintering temperatures rather than the pure ZnO and Fe2O3. The enhanced antibacterial properties were likely caused by the spongy ZnO/ZnFe2O4 heterostructures, improving the probability of photoinduced charge separation and broadening the visible-light absorption.

[The Role of Postoperative Radiotherapy on Stage N2 Non-small Cell Lung Cancer.].

BACKGROUND: The clinical value of postoperative radiotherapy (PORT) in stage N2 nonsmall-cell lung cancer (NSCLC) is controversy. The aim of this study is to analyze the efficacy of PORT in subgroup of stage N2 NSCLC, which can help clinicians to choose proper patients for PORT. METHODS: Clinical data of 359 patients with stage N2 NSCLC treated with radical surgery between Mar. 2000 and Jul. 2005 were retrospectively reviewed. Two hundred and seven patients received adjuvant chemotherapy and one hundred and four patients received adjuvant radiotherapy. First, the group of patients were analyzed to evaluate the factors affecting the overall survival. The all patients were divided based on tumor size and the number of lymph node metastasis station (single station or multiple station) so as to evaluate the role of PORT. The endpoint was overall survival (OS) and local recurrence-free survival (LRFS). Kaplan-Meier method was used to calculate the OS, LRFS and Log-rank was used to compare the difference in OS and LRFS between different groups. RESULTS: The median duration of follow-up was 2.3 years. 224 patients died. The median survival was 1.5 years and 1, 3, 5-year survival were 78%, 38% and 26%. Univariate analysis showed tumor size, the number of lymph node metastasis station and PORT were correlated with OS. Among patients, 5-year survival rates in PORT and non-PORT were 29% and 24% (P=0.047) respectively. In subgroups, PORT was related with high survival in patients with multiple station N2 compared to non-PORT: 36% vs 20% (P=0.013) and 33% vs 15% (P=0.002) in patients in patients with tumor size>3 cm. Also, it was related with low local recurrence compared to non-PORT: 65% vs 48% (P=0.006) and 62% vs 48% (P=0.033). CONCLUSIONS: PORT can improve overall survival for N2 NSCLC, especially the patients with the factors as follows: tumor size>3 cm and multiple station N2 can benefit from PORT more or less.