Expression of GnT-III decreases chemoresistance via negatively regulating P-glycoprotein expression: Involvement of the TNFR2-NF-κB signaling pathway.

The phenomenon of multidrug resistance (MDR) is called chemoresistance with respect to the treatment of cancer, and it continues to be a major challenge. The role of N-glycosylation in chemoresistance, however, remains poorly understood. Here, we established a traditional model for adriamycin resistance in K562 cells, which are also known as K562/adriamycin-resistant (ADR) cells. Lectin blot, mass spectrometry, and RT-PCR analysis showed that the expression levels of N-acetylglucosaminyltransferase III (GnT-III) mRNA and its products, bisected N-glycans, are significantly decreased in K562/ADR cells, compared with the levels in parent K562 cells. By contrast, the expression levels of both P-glycoprotein (P-gp) and its intracellular key regulator, NF-κB signaling, are significantly increased in K562/ADR cells. These upregulations were sufficiently suppressed by the overexpression of GnT-III in K562/ADR cells. We found that the expression of GnT-III consistently decreased chemoresistance for doxorubicin and dasatinib, as well as activation of the NF-κB pathway by tumor necrosis factor (TNF) α, which binds to two structurally distinct glycoproteins, TNF receptor 1 (TNFR1) and TNF receptor 2 (TNFR2), on the cell surface. Interestingly, our immunoprecipitation analysis revealed that only TNFR2, but not TNFR1, contains bisected N-glycans. The lack of GnT-III strongly induced TNFR2's autotrimerization without ligand stimulation, which was rescued by the overexpression of GnT-III in K562/ADR cells. Furthermore, the deficiency of TNFR2 suppressed P-gp expression while it increased GnT-III expression. Taken together, these results clearly show that GnT-III negatively regulates chemoresistance via the suppression of P-gp expression, which is regulated by the TNFR2-NF/κB signaling pathway.

Taurine promotes B-cell activation by interaction with the VH /VL framework regions of B-cell receptor.

Taurine (Tau) is a special sulphur-containing amino acid and has been widely used as a dietary supplement. Although Tau exists in lymphocytes in large quantities, the physiological significance of Tau to modulate human immunity is unknown. In the present study, we first found that Tau regulates the B-cell receptor (BCR)-mediated signal transduction and induces the B cells activation. The IgG production of mice after ovalbumin immunization was also increased by Tau administration. Moreover, the isothermal titration calorimetry and surface plasmon resonance analysis have shown that Tau specifically bound to the IgG2a-BCR. The Tau could bind to IgG F(ab')2 regions via fluorescence spectroscopy analysis. In the molecular docking analysis, Tau bound to the framework regions (FRs) of variable region of the heavy chains (VH ) and in the light chains (VL ) of IgG2a-BCR. Our results suggested that Tau could improve the activation of B cells by interaction with the VH /VL FRs of BCR.

O-GlcNAcylation regulates ß1,4-GlcNAc-branched N-glycan biosynthesis via the OGT/SLC35A3/GnT-IV axis.

N-Linked glycosylation and O-linked N-acetylglucosamine (O-GlcNAc) are important protein post-translational modifications that are orchestrated by a diverse set of gene products. Thus far, the relationship between these two types of glycosylation has remained elusive, and it is unclear whether one influences the other via UDP-GlcNAc, which is a common donor substrate. Theoretically, a decrease in O-GlcNAcylation may increase the products of GlcNAc-branched N-glycans. In this study, via examination by lectin blotting, HPLC, and mass spectrometry analysis, however, we found that the amounts of GlcNAc-branched tri-antennary N-glycans catalyzed by N-acetylglucosaminyltransferase IV (GnT-IV) and tetra-antennary N-glycans were significantly decreased in O-GlcNAc transferase knockdown cells (OGT-KD) compared with those in wild type cells. We examined this specific alteration by focusing on SLC35A3, which is the main UDP-GlcNAc transporter in mammals that is believed to modulate GnT-IV activation. It is interesting that a deficiency of SLC35A3 specifically leads to a decrease in the amounts of GlcNAc-branched tri- and tetra-antennary N-glycans. Furthermore, co-immunoprecipitation experiments have shown that SLC35A3 interacts with GnT-IV, but not with N-acetylglucosaminyltransferase V. Western blot and chemoenzymatic labeling assay have confirmed that OGT modifies SLC35A3 and that O-GlcNAcylation contributes to its stability. Furthermore, we found that SLC35A3-KO enhances cell spreading and suppresses both cell migration and cell proliferation, which is similar to the phenomena observed in the OGT-KD cells. Taken together, these data are the first to demonstrate that O-GlcNAcylation specifically governs the biosynthesis of tri- and tetra-antennary N-glycans via the OGT-SLC35A3-GnT-IV axis.

Cell membrane-camouflaged inorganic nanoparticles for cancer therapy.

Inorganic nanoparticles (INPs) have been paid great attention in the field of oncology in recent past years since they have enormous potential in drug delivery, gene delivery, photodynamic therapy (PDT), photothermal therapy (PTT), bio-imaging, driven motion, etc. To overcome the innate limitations of the conventional INPs, such as fast elimination by the immune system, low accumulation in tumor sites, and severe toxicity to the organism, great efforts have recently been made to modify naked INPs, facilitating their clinical application. Taking inspiration from nature, considerable researchers have exploited cell membrane-camouflaged INPs (CMCINPs) by coating various cell membranes onto INPs. CMCINPs naturally inherit the surface adhesive molecules, receptors, and functional proteins from the original cell membrane, making them versatile as the natural cells. In order to give a timely and representative review on this rapidly developing research subject, we highlighted recent advances in CMCINPs with superior unique merits of various INPs and natural cell membranes for cancer therapy applications. The opportunity and obstacles of CMCINPs for clinical translation were also discussed. The review is expected to assist researchers in better eliciting the effect of CMCINPs for the management of tumors and may catalyze breakthroughs in this area.

Core fucosylation of copper transporter 1 plays a crucial role in cisplatin-resistance of epithelial ovarian cancer by regulating drug uptake.

Core fucosylation catalyzed by core fucosyltransferase (Fut8) contributes to the progressions of epithelial ovarian cancer (EOC). Copper transporter 1 (CTR1), which contains one N-glycan on Asn15 , mediates cellular transport of cisplatin (cDDP), and plays an important role in the process of cDDP-resistance in EOC. In the present study, we found that the core fucosylation level elevated significantly in the sera of cDDP-treated EOC patients. The in vitro assays also indicate that core fucosylation of CTR1 was significantly upregulated in cDDP-resistant A2780CP cells compared to the cDDP-sensitive A2780S cells. Intriguingly, the hyper core fucosylation suppressed the CTR1-cDDP interactions and cDDP-uptake into A2780CP cells. Conversely, contrast to the Fut8+/+ mouse ovarian epithelial cells, the Fut8-deleted (Fut8-/- ) cells obviously showed higher cDDP-uptake. Furthermore, the recovered core fucosylation induced the suppression of cDDP-uptake in Fut8-restored ovarian epithelial cells. In addition, the core fucosylation could regulate the phosphorylation of cDDP-resistance-associated molecules, such as AKT, ERK, JNK, and mTOR. Our findings suggest that the core fucosylation of CTR1 plays an important role in the cellular cDDP-uptake and thus provide new strategies for improving the outcome of cDDP based chemotherapy of EOC.

Effect of Additives on Tribological Performance of Magnetorheological Fluids.

In this study, nano-diamond (ND) and MoS2 powder are used as additives in a carbonyl iron-based magnetorheological fluid (MRF) to improve its tribological performance. MRFs are prepared by dispersing 35 wt.% of CI particles in silicone oil and adding different proportions (0, 1, 3, or 5 wt.%) of ND and MoS2 additives. Seven kinds of MRFs are made and tested using reciprocating friction and wear tester under different normal loads, and then the friction characteristics are evaluated by analyzing the experimental results. The morphological properties of MRFs and contacting surfaces before and after the tests are also observed using a scanning electron microscope and analyzed via energy-dispersive X-ray spectroscopy. The results show that the appropriate weight percentage of MoS2 additives may decrease the friction coefficient and wear zone. It is also demonstrated from detailed analyses of worn surfaces that the wear mechanism is influenced not only by additives, but also by the applied normal load and magnetic field strength.

Core fucosylation regulates the ovarian response via FSH receptor during follicular development.

INTRODUCTION: Ovarian low response to follicle-stimulating hormone (FSH) causes infertility featuring hypergonadotropic hypogonadism, ovarian failure, and/or defective ovarian response. OBJECTIVES: N-glycosylation is essential for FSH receptor (FSHR). Core fucosylation catalyzed by fucosyltransferase 8 (FUT8) is the most common N-glycosylation. Core fucosylation level changes between individuals and plays important roles in multiple physiological and pathological conditions. This study aims to elucidate the significance of FUT8 to modulate FSHR function in female fertility. METHODS: Samples from patients classified as poor ovary responders (PORs) were detected with lectin blot and real-time PCR. Fut8 gene knockout (Fut8-/-) mice and FUT8-knockdown human granulosa cell line (KGN-KD) were established and in vitro fertilization (IVF) assay, western blot, molecular interaction, immunofluorescence and immunoprecipitation were applied. RESULTS: Core fucosylation is indispensable for oocyte and follicular development. FSHR is a highly core-fucosylated glycoprotein. Loss of core fucosylation suppressed binding of FSHR to FSH, and attenuated FSHR downstream signaling in granulosa cells. Transcriptomic analysis revealed the downregulation of several transcripts crucial for oocyte meiotic progression and preimplantation development in Fut8-/- mice and in POR patients. Furthermore, loss of FUT8 inhibited the interaction between granulosa cells and oocytes, reduced transzonal projection (TZP) formation and caused poor developmental competence of oocytes after fertilization in vitro. While L-fucose administration increased the core fucosylation of FSHR, and its sensitivity to FSH. CONCLUSION: This study first reveals a significant presence of core fucosylation in female fertility control. Decreased fucosylation on FSHR reduces the interaction of FSH-FSHR and subsequent signaling, which is a feature of the POR patients. Our results suggest that core fucosylation controls oocyte and follicular development via the FSH/FSHR pathway and is essential for female fertility in mammals.

Ablation of the GDP-fucose transporter suppresses lung cancer cell proliferation and migration by reducing expression of PD-L1.

Fucosylation, an important post-translational modification, is closely related to the development of tumors. In the microenvironment of lung cancer, expression of PD-L1 and fucosylation is abnormally upregulated. However, the correlation between PD-L1 expression and its fucosylation in lung adenocarcinoma (LUAD) remains unclear. The GDP-fucose transporter (GFT) is a key molecule in cellular fucosylation. To explore the correlation between fucosylation and PD-L1 expression, we knocked out the GFT-encoding gene SLC35C1 in mouse Lewis lung adenocarcinoma cells and in human H1299 lung adenocarcinoma cells. Loss of SLC35C1 impaired the phosphorylation of EGFR and its downstream target ERK. Moreover, loss of SLC35C1 up-regulated the expression of ß-TrCP, a PD-L1 E3 ligase, thereby promoting the ubiquitination of PD-L1 and its subsequent degradation. The down-regulated expression of PD-L1 leads to a decline in lung cancer cell proliferation and migration. These results suggest that fucosylation partially influences LUAD tumorigenesis by regulating PD-L1 expression.

Engineering white blood cell membrane-camouflaged nanocarriers for inflammation-related therapeutics.

White blood cells (WBCs) play essential roles against inflammatory disorders, bacterial infections, and cancers. Inspired by nature, WBC membrane-camouflaged nanocarriers (WBC-NCs) have been developed to mimic the "dynamic" functions of WBCs, such as transendothelial migration, adhesion to injured blood vessels, etc, which make them promising for diverse medical applications. WBC-NCs inherit the cell membrane antigens of WBCs, while still exhibiting the robust inflammation-related therapeutic potential of synthetic nanocarriers with excellent (bio)physicochemical performance. This review summarizes the proposed concept of cell membrane engineering, which utilizes physical engineering, chemical modification, and biological functionalization technologies to endow the natural cell membrane with abundant functionalities. In addition, it highlights the recent progress and applications of WBC-NCs for inflammation targeting, biological neutralization, and immune modulation. Finally, the challenges and opportunities in realizing the full potential of WBC-NCs for the manipulation of inflammation-related therapeutics are discussed.

Hydrogel-Anchored Fe-Based Amorphous Coatings with Integrated Antifouling and Anticorrosion Functionality.

Biofouling and corrosion of underwater equipment induced by marine organisms have become major issues in the marine industry. The superior corrosion resistance of Fe-based amorphous coatings makes them suitable for marine applications; however, they have a poor antifouling ability. In this work, a hydrogel-anchored amorphous (HAM) coating with satisfactory antifouling and anticorrosion performance is designed, utilizing an interfacial engineering strategy involving micropatterning, surface hydroxylation, and a dopamine intermediate layer to increase the adhesion strength between the hydrogel layer and the amorphous coating. The as-obtained HAM coating exhibits exceptional antifouling properties, achieving 99.8% resistance to algae, 100% resistance to mussels, and excellent biocorrosion resistance against Pseudomonas aeruginosa. Antifouling and anticorrosion performance of the HAM coating was also explored by conducting a marine field test in the East China Sea, and no signs of corrosion and fouling are observed after 1 month of immersion. It is revealed that the outstanding antifouling properties stem from the killing-resisting-camouflaging trinity that resists organism attachment across different length scales, and the excellent anticorrosion performance originates from the remarkable barrier of the amorphous coating against Cl- ion diffusion and microbe-induced biocorrosion. This work presents a novel methodology for designing marine protective coating with excellent antifouling and anticorrosion properties.

Core fucosylation and its roles in gastrointestinal glycoimmunology.

Glycosylation is a common post-translational modification in eukaryotic cells. It is involved in the production of many biologically active glycoproteins and the regulation of protein structure and function. Core fucosylation plays a vital role in the immune response. Most immune system molecules are core fucosylated glycoproteins such as complements, cluster differentiation antigens, immunoglobulins, cytokines, major histocompatibility complex molecules, adhesion molecules, and immune molecule synthesis-related transcription factors. These core fucosylated glycoproteins play important roles in antigen recognition and clearance, cell adhesion, lymphocyte activation, apoptosis, signal transduction, and endocytosis. Core fucosylation is dominated by fucosyltransferase 8 (Fut8), which catalyzes the addition of α-1,6-fucose to the innermost GlcNAc residue of N-glycans. Fut8 is involved in humoral, cellular, and mucosal immunity. Tumor immunology is associated with aberrant core fucosylation. Here, we summarize the roles and potential modulatory mechanisms of Fut8 in various immune processes of the gastrointestinal system.

α1,6-Fucosyltransferase contributes to cell migration and proliferation as well as to cancer stemness features in pancreatic carcinoma.

BACKGROUND: Pancreatic carcinoma is one of the deadliest malignant diseases, in which the increased expression of α1,6-fucosyltransferase (FUT8), a sole enzyme responsible for catalyzing core fucosylation, has been reported. However, its pathological roles and regulatory mechanisms remain largely unknown. Here, we use two pancreatic adenocarcinoma cell lines, MIA PaCa-2 and PANC-1 cells, as cell models, to explore the relationship of FUT8 with the malignant transformation of PDAC. METHODS: FUT8 knockout (FUT8-KO) cells were established by the CRISPR/Cas9 system. Cell migration was analyzed by transwell and wound-healing assays. Cell proliferation was examined by MTT and colony-formation assays. Cancer stemness markers and spheroid formations were used to analyzed cancer stemness features. RESULTS: Deficiency of FUT8 inhibited cell migration and proliferation in both MIA PaCa-2 and PANC-1 cells compared with wild-type cells. Moreover, the expression levels of cancer stemness markers such as EpCAM, CXCR4, c-Met, and CD133 were decreased in the FUT8-KO cells compared with wild-type cells. Also, the spheroid formations in the KO cells were loose and unstable, which could be reversed by restoration with FUT8 gene in the KO cells. Additionally, FUT8-KO increased the chemosensitivity to gemcitabine, which is the first-line therapy for advanced pancreatic cancer. CONCLUSIONS: FUT8-KO reduced the cell proliferation and migration. Our results are the first to suggest that the expression of FUT8 is involved in regulating the stemness features of pancreatic cancer cells. GENERAL SIGNIFICANCE: FUT8 could provide novel insights for the treatment of pancreatic carcinoma.

Annular Surface Micromachining of Titanium Tubes Using a Magnetorheological Polishing Technique.

In this study, a novel magnetorheological (MR) polishing device under a compound magnetic field is designed to achieve microlevel polishing of the titanium tubes. The polishing process is realized by combining the rotation motion of the tube and the reciprocating linear motion of the polishing head. Two types of excitation equipment for generating an appropriate compound magnetic field are outlined. A series of experiments are conducted to systematically investigate the effect of compound magnetic field strength, rotation speed, and type and concentration of abrasive particles on the polishing performance delivered by the designed device. The experiments were carried out through controlling variables. Before and after the experiment, the surface roughness in the polished area of the workpiece is measured, and the influence of the independent variable on the polishing effect is judged by a changing rule of surface roughness so as to obtain a better parameter about compound magnetic field strength, concentration of abrasive particles, etc. It is shown from experimental results that diamond abrasive particles are appropriate for fine finishing the internal surface of the titanium-alloy tube. It is also identified that the polishing performance is excellent at high magnetic field strength, fast rotation speed, and high abrasive-particle concentration.

Microstructure Simulation and Constitutive Modelling of Magnetorheological Fluids Based on the Hexagonal Close-packed Structure.

This paper presents a new constitutive model of high particles concentrated magnetorheological fluids (MRFs) that is based on the hexagonal close-packed structure, which can reflect the micro-structures of the particles under the magnetic field. Firstly, the particle dynamic simulations for the forces sustained by carbonyl iron powder (CIP) particles of MRFs are performed in order to investigate the particles chain-forming process at different time nodes. Subsequently, according to the force analyses, a hexagonal close-packed structure, which differs from the existing single-chain structure and body-cantered cubic structure, is adopted to formulate a constitutive model of MRFs with high concentration of the magnetic-responsive particles. Several experiments are performed while considering crucial factors that influence on the chain-forming mechanism and, hence, change the field-dependent shear yield stress in order to validate the proposed model. These factors include the magnetic induction intensity, volume fraction and radius of CIP particles, and surfactant coating thickness. It is shown that the proposed modeling approach can predict the field-dependent shear yield stress much better than the single-chain model. In addition, it is identified that the shear yield stress is increased as the particle volume fraction increases and surfactant coating thickness decreases. It is believed that the proposed constitutive model can be effectively used to estimate the field-dependent shear yield stress of MRFs with a high concentration of iron particles.

A novel immunochromatographic strips assay for rapid and simple detection of systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a complex multi-system autoimmune disease. Detection of anti-nuclear antibodies (ANA) is fundamental for the diagnosis of SLE. In the present study, we found that the level of core fucosylation catalyzed by α1,6-fucosyltransferase (Fut8) is markedly up-regulated on immunoglobulin G (IgG) in the sera of SLE patients detected by Aspergillus oryzae lectin (AOL) blot. In sandwich Dot enzyme-linked immunosorbent assay (Dot-ELISA), the core fucosylation level was also found significantly increased in the sera from SLE patients with a higher ANA titer. To establish a rapid and sensitive laboratory test for the diagnosis of SLE, we prokaryotically expressed AOL and C3-D1-C3-D2-C3 of protein G (SpG3), and generate AOL-conjugated colloid gold immunochromatographic strips (ICS). The detection limit of core fucosylated IgG was 10 µg/mL for AOL-conjugated colloid gold ICS. As well as indirect immunofluorescence, the AOL-conjugated colloid gold ICS showed reliable results in the serum of 39 SLE patients. Our results indicated that the AOL-conjugated colloid gold ICS could serve as a rapid test for the detection of ANA and suspected cases of SLE.

Association of TP53 codon 72 genotype polymorphism and environmental factors with esophageal squamous cell carcinoma in the Mongolian population of the Chinese region of Inner Mongolia.

The present study analyzed the association of tumor protein p53 (TP53) Pro72Arg polymorphism with esophageal squamous cell carcinoma (ESCC) in the Mongolian population of Tongliao (Inner Mongolia, China). Restriction fragment length polymorphism-polymerase chain reaction was used to detect the genotype distribution of TP53 Pro72Arg polymorphism in 100 patients with ESCC and 50 healthy controls from the same population. Besides, the correlation between ESCC in Mongolian patients and various factors such as age, sex, cigarette smoking and alcohol consumption was analyzed. χ2 test revealed significant associations between alcohol consumption (P=0.00006) and cigarette smoking (P=0.00076) and ESCC in Mongolian patients. Notably, the Pro72 allele was significantly enriched in patients with ESCC compared with its abundance in the healthy control group, and the genotype of Pro/Arg on p53 codon 72 was confirmed to exhibit a significant correlation with ESCC in Mongolian patients. The present study demonstrated that alcohol drinking and cigarette smoking were risk factors for ESCC in the Mongolian population. Mongolian patients who carry the partocular genotype of Arg/Pro or Pro/Pro on p53 codon 72 may be more likely to develop ESCC. Compared with the p53 codon 72 genotype Arg/Arg, the TP53 Pro72 allele increased the risk of ESCC in Mongolian patients by 1.659-fold.

Spontaneous epidural hematoma complicated by systemic lupus erythematosus: one case report.

In this study, an unusual case of intracranial hemorrhage is presented. It is spontaneous epidural hemorrhage, the complication of Systemic Lupus Erythematosus. In this case, a 29-year-old female presented with vomiting and continuous headache and computed tomography revealed the right frontal parietal and temporal epidural hematoma. The patient had been diagnosed SLE one year ago. Together, these observations indicated that the patient needs a surgery to reduce the intracranial pressure. Following surgery, the symptoms were eradicated but 7 hours later after surgery the review head CT showed that left epidural hemorrhage. According to the surgery index, we decided to give the patient non-operative treatment and the intracranial hemorrhage was under control. We thought this was the surgery complication but it's not. On the eighth day after surgery, the patient had a sudden headache with vomiting again and head CT revealed that left epidural hemorrhage. But this time we just gave non-operative treatment and especially added the dose of glucocorticosteroid. 12 days later, the patient's symptoms were under control and she was discharged from hospital. We also reviewed the literature about spontaneous epidural hemorrhage and bilateral epidural hematomas.