Establishment and characterization of a fin tissue cell line derived from silver pomfret, Pampus argenteus.

A cell line (PaF) derived from the fin tissue of silver pomfret (Pampus argenteus) was established and characterized in this study. The cell line has been subcultured for more than 50 times in Dulbecco's modified Eagle's medium (DMEM) containing 15% foetal bovine serum (FBS) since the initial primary culture. PaF cells grew well at temperatures from 24°C to 28°C in DMEM supplemented with 15% FBS. Partial amplification and sequence analysis of the cytochrome B gene indicated that PaF originated from silver pomfret. Cytogenetic analysis demonstrated that the modal chromosome number was 48. A significant cytopathic effect was observed in PaF cells during viral haemorrhagic septicaemia virus (VHSV) infection, and the VHSV replication was confirmed by qRT-PCR and viral titre assays. In contrast, PaF cells were resistant to red-spotted grouper nervous necrosis virus infection. Moreover, PaF cells could respond to VHSV and lipopolysaccharide treatments, as indicated by the expression of immune-related genes, TLR5 and TLR9. In conclusion, the establishment of PaF cell line will provide an appropriate in vitro tool for the study of mechanisms of pathogen-silver pomfret interaction.

Enhanced thermoelectric performance of Cu3SbS4 flower-like hierarchical architectures composed of Cl doped nanoflakes via an in situ generated CuS template.

In this work, Cu3SbS4 hierarchical flower-like microspheres composed of chlorine (Cl-)-doped Cu3SbS4 nanoflakes are realized via a one pot solvothermal ion exchange reaction. The kinetic factors including the duration time, the ratio of source materials, and the KOH concentration, are systematically investigated. Using a suite of analytical techniques, including SEM, XRD and FTIR, the mechanism of the two stage in situ chemical transformation of CuS flower-like microspheres consisting of nanoflake intermediates to the target product Cu3SbS4 is elucidated. The difference in solubility between reactants and products (Ksp(CuS) > Ksp(CuSbSx)) determines that the ion-exchange reaction from transition binary to ternary metal chalcogenides is favorable under the impetus of a thermodynamic driving force. In addition, the optical and enhanced thermoelectric transport properties are investigated. The results revealed that Cl-doped Cu3SbS4 exhibited an improved power factor, which was 8 times higher than that of undoped Cu3SbS4 at 500 K. The current study not only provides a facile and economical way to synthesize high-quality Cl-doped Cu-Sb-S three dimensional (3D) hierarchical nanostructures, but also opens up a new route for preparation of other I-V-VI multicomponent chalcogenide NCs, such as Cu-Bi-S and Cu-Pb-S systems, which would be difficult to obtain otherwise.

Comparison of seven CD19 CAR designs in engineering NK cells for enhancing anti-tumour activity.

Chimeric antigen receptor-natural killer (CAR-NK) cell therapy is emerging as a promising cancer treatment, with notable safety and source diversity benefits over CAR-T cells. This study focused on optimizing CAR constructs for NK cells to maximize their therapeutic potential. We designed seven CD19 CAR constructs and expressed them in NK cells using a retroviral system, assessing their tumour-killing efficacy and persistence. Results showed all constructs enhanced tumour-killing and prolonged survival in tumour-bearing mice. In particular, CAR1 (CD8 TMD-CD3ζ SD)-NK cells showed superior efficacy in treating tumour-bearing animals and exhibited enhanced persistence when combined with OX40 co-stimulatory domain. Of note, CAR1-NK cells were most effective at lower effector-to-target ratios, while CAR4 (CD8 TMD-OX40 CD- FcεRIγ SD) compromised NK cell expansion ability. Superior survival rates were noted in mice treated with CAR1-, CAR2 (CD8 TMD- FcεRIγ SD)-, CAR3 (CD8 TMD-OX40 CD- CD3ζ SD)- and CAR4-NK cells over those treated with CAR5 (CD28 TMD- FcεRIγ SD)-, CAR6 (CD8 TMD-4-1BB CD-CD3ζ 1-ITAM SD)- and CAR7 (CD8 TMD-OX40 CD-CD3ζ 1-ITAM SD)-NK cells, with CAR5-NK cells showing the weakest anti-tumour activity. Increased expression of exhaustion markers, especially in CAR7-NK cells, suggests that combining CAR-NK cells with immune checkpoint inhibitors might improve anti-tumour outcomes. These findings provide crucial insights for developing CAR-NK cell products for clinical applications.

Characterization of a novel polysaccharide from Arca subcrenata and its immunoregulatory activities in vitro and in vivo.

Arca subcrenata is an economical edible shellfish. A novel water-soluble α-D-glucan (ASPG-1) with a molecular weight of 2.56 × 106 Da was purified and characterized from A. subcrenata. Its structure was characterized as a repeating unit consisting of α-D-Glcp, (1 → 6)-α-D-Glcp and (1 → 4,6)-α-D-Glcp. ASPG-1 exerted potent immunoregulatory activity by promoting the viability of splenic lymphocytes. Moreover, it enhanced pinocytic capacity, and promoted the secretion of NO and cytokines in RAW264.7 cells. The immunomodulatory mechanism of ASPG-1 involved the activation of the TLR4-MAPK/Akt-NF-κB signaling pathway. ASPG-1 inhibited tumor growth in 4T1 breast cancer mice and its combination with doxorubicin increased antitumor efficacy. The ASPG-1 combination with DOX-treated group (64.8%) showed an improved tumor inhibition rate compared to that of the DOX-treated group (53.3%). The antitumor mechanism of ASPG-1 may involve an enhancement of the immune response of mice to tumors. These results indicated that ASPG-1 could be developed as a potential adjuvant in tumor immunotherapy.

Structural characterization and immunoregulatory activity of a novel acidic polysaccharide from Scapharca subcrenata.

A novel acidic polysaccharide named SSPA50-1 was isolated from Scapharca subcrenata using a simulated gastric fluid extraction method. SSPA50-1 is a heteropolysaccharide with an average molecular weight of 44.7 kDa that is composed of galacturonic acid, glucose, galactose, mannose, ribose, rhamnose, fucose, xylose and arabinose at a molar ratio of 1.00:5.40:9.04:3.10:1.59:4.01:2.10:2.21:2.28. The structural characterization based on the methylation and 1D/2D NMR analyses indicated that SSPA50-1 is composed of →3)-ß-L-Rhap-(1→, →3)-ß-L-2-O-Me-Fucp-(1→, →2)-α-D-Xylp-(1→, →5)-α-L-Araf-(1→, →3)-ß-D-Galp-(1→, →6)-α-D-Glcp-(1→, →3,4)-ß-D-Manp-(1→, →3,4)-ß-D-Galp-(1→, ß-D-Ribf-(1→, α-D-Glcp-(1→, and α-D-GalAp6Me-(1→. Furthermore, SSPA50-1 possessed potent immunoregulatory activity by enhancing the phagocytosis and NO, iNOS, TNF-α and IL-6 secretion capacity of RAW264.7 cells. Otherwise, SSPA50-1 significantly promoted the proliferation of splenic lymphocytes and RAW264.7 macrophages. These results indicated that SSPA50-1 could be developed as a potential ingredient for immunostimulatory agents.

Arteannuin B Enhances the Effectiveness of Cisplatin in Non-Small Cell Lung Cancer by Regulating Connexin 43 and MAPK Pathway.

Cisplatin (DDP)-based chemotherapy is the first-line regimen for advanced non-small cell lung cancer (NSCLC) patients. However, advanced NSCLC patients may have innate resistance to DDP or develop resistance during DDP treatment. We investigated a natural compound, arteannuin B (Art B), for its potential effects on DDP resistance in NSCLC. Art B was isolated from Artemisia annua by chromatographic purification and spectral elucidation. The activities of Art B on DDP-mediated effects were examined using in vitro and in vivo assays. We observed significant correlations in T stage, clinical stage, chemotherapy resistance and poor survival of NSCLC patients with low Cx43 expression. Art B enhanced the effectiveness of cisplatin by increasing Cx43 expression in normal and DDP-resistant NSCLC cells. Art B also increased DDP uptake through up-regulating Cx43. The combination of DDP and Art B showed better therapeutic effect than individual treatments both in vitro and in vivo. Art B increased intracellular Fe[Formula: see text] level, promoted calcium influx, and activated gap junction and MAPK pathways, which might contribute to Art B-mediated effects. Art B may serve as a new drug candidate to enhance the antitumor effect of DDP on NSCLC.

Lateral plate mesoderm cell-based organoid system for NK cell regeneration from human pluripotent stem cells.

Human pluripotent stem cell (hPSC)-induced NK (iNK) cells are a source of off-the-shelf cell products for universal immune therapy. Conventional methods for iNK cell regeneration from hPSCs include embryoid body (EB) formation and feeder-based expansion steps, which are time-consuming and cause instability and high costs of manufacturing. Here, we develop an EB-free, organoid aggregate method for NK cell regeneration from hPSCs. In a short time-window of 27-day induction, millions of hPSC input can output over billions of iNK cells without the necessity of NK cell expansion feeders. The iNK cells highly express classical toxic granule proteins, apoptosis-inducing ligands, as well as abundant activating and inhibitory receptors. Functionally, the iNK cells eradicate human tumor cells via mechanisms of direct cytotoxicity, apoptosis, and antibody-dependent cellular cytotoxicity. This study provides a reliable scale-up method for regenerating human NK cells from hPSCs, which promotes the universal availability of NK cell products for immune therapy.

Purification and characterization of a novel mixed-linkage α,ß-d-glucan from Arca subcrenata and its immunoregulatory activity.

Arca subcrenata Lischke is a seafood with high nutritional value. In this study, we purified and characterized a novel water-soluble polysaccharide (ASPG-2) from Arca subcrenata with significant immunoregulatory effects and no apparent cell toxicity. ASPG-2 is a class of mixed-linkage α,ß-d-glucan backbones with α-linked side chains with a molecular weight of 4.39 × 105 Da. Its structure was characterized as a repeating unit consisting of (1 → 3)-ß-d-Glcp, (1 → 4)-α-d-Glcp, (1 → 4,6)-α-d-Glcp and (1 → 6)-α-d-Glcp. Using mouse RAW264.7 macrophages, we demonstrated that ASPG-2 exerted marked immunoregulatory effects by promoting the secretion of NO and increasing the phagocytosis of RAW264.7 cells in vitro. Moreover, flow cytometry analysis of the expression of the cell surface molecule CD86 revealed that ASPG-2 could polarize RAW264.7 cells into the M1 type. The immunomodulatory mechanism of ASPG-2 in macrophages was associated with the activation of the TLR4-MAPK/Akt-NF-κB signalling pathways. These results indicated that ASPG-2 might be researched and developed as a potential immunomodulatory agent or health product from marine organisms.