Blockade of IL-6 inhibits tumor immune evasion and improves anti-PD-1 immunotherapy.

Long-standing inflammatory bowel disease predisposes to the development of colorectal cancer (CRC). Interleukin (IL) -6, a pivotal link between chronic inflammation and tumor progression, has recently been recognized as a potential therapeutic target. The effect of IL-6 on proliferation and metastasis of CRC by activating the STAT3 pathway has been widely demonstrated in recent years, but few on mediating tumor immune evasion. In this study, we found that IL-6 was remarkably overexpressed in CRC and its elevation was associated with a poor prognosis. We studied CRC tumorigenesis in vivo by inoculating MC38 tumors and induced-CRC model via AOM/DSS (azoxymethane/dextransulfate sodium) in IL-6 deficient (IL-6-/-) and wild-type (WT) mice and found that IL-6-/- mice were less susceptible to develop tumors, compared to WT mice. We detected CD8+ T cells via immunofluorescence and found they exhibit high expression in tumor of IL-6-/- mice. High level of IL-6 was found in colitis model, with down-regulation of MHC-I molecules. In in vitro experiments, we found that IL-6 may act as a negative regulator in IFNγ-STAT1-MHC-I signaling. In addition, vivo trials also confirmed that MHC-I mRNA level was negatively related to the existence of IL-6. Furthermore, the blockade of IL-6 also activated CD8+T-cell accumulation and led to the high PD-L1 expression in CRC, which can sensitize animals to anti-PD-1 therapy. Our study provides a research basis for the significant role of IL-6 in tumor evasion and highlights a novel target to improve the efficacy of immunotherapy.

STOML2 interacts with PHB through activating MAPK signaling pathway to promote colorectal Cancer proliferation.

BACKGROUND: Highly expressed STOML2 has been reported in a variety of cancers, yet few have detailed its function and regulatory mechanism. This research aims to reveal regulatory mechanism of STOML2 and to provide evidence for clinical therapeutics, via exploration of its role in colorectal cancer, and identification of its interacting protein. METHODS: Expression level of STOML2 in normal colon and CRC tissue from biobank in Nanfang Hospital was detected by pathologic methods. The malignant proliferation of CRC induced by STOML2 was validated via gain-of-function and loss-of-function experiments, with novel techniques applied, such as organoid culture, orthotopic model and endoscopy monitoring. Yeast two-hybrid assay screened interacting proteins of STOML2, followed by bioinformatics analysis to predict biological function and signaling pathway of candidate proteins. Target protein with most functional similarity to STOML2 was validated with co-immunoprecipitation, and immunofluorescence were conducted to co-localize STOML2 and PHB. Pathway regulated by STOML2 was detected with immunoblotting, and subsequent experimental therapy was conducted with RAF inhibitor Sorafenib. RESULTS: STOML2 was significantly overexpressed in colorectal cancer and its elevation was associated with unfavorable prognosis. Knockdown of STOML2 suppressed proliferation of colorectal cancer, thus attenuated subcutaneous and orthotopic tumor growth, while overexpressed STOML2 promoted proliferation in cell lines and organoids. A list of 13 interacting proteins was screened out by yeast two-hybrid assay. DTYMK and PHB were identified to be most similar to STOML2 according to bioinformatics in terms of biological process and signaling pathways; however, co-immunoprecipitation confirmed interaction between STOML2 and PHB, rather than DTYMK, despite its highest rank in previous analysis. Co-localization between STOML2 and PHB was confirmed in cell lines and tissue level. Furthermore, knockdown of STOML2 downregulated phosphorylation of RAF1, MEK1/2, and ERK1/2 on the MAPK signaling pathway, indicating common pathway activated by STOML2 and PHB in colorectal cancer proliferation. CONCLUSIONS: This study demonstrated that in colorectal cancer, STOML2 expression is elevated and interacts with PHB through activating MAPK signaling pathway, to promote proliferation both in vitro and in vivo. In addition, combination of screening assay and bioinformatics marks great significance in methodology to explore regulatory mechanism of protein of interest.

Protein-reduced gold nanoparticles mixed with gentamicin sulfate and loaded into konjac/gelatin sponge heal wounds and kill drug-resistant bacteria.

Timely antibacterial treatment of wounds reduces the probability of wound infection and promotes wound healing. However, the materials used to treat wounds often fail to provide both sterilization (especially for super bacteria) and moisture, and some may even cause secondary injury to the wound. In this study, gold nanoparticles (Au NPs) of average grain diameter of 3 ± 1 nm were prepared using egg white as the reductant. These particles showed no aggregation and pink fluorescence. Au NPs were mixed with gentamicin sulfate (GS) and loaded into a mixture of konjac glucomannan (KGM) and gelatin as wound dressing (KGM/Gelatin@Au NPs/GS). Antibacterial experiments showed that the Au NPs amplified the antibacterial activity of GS; Au NPs/GS efficiently eliminated bacteria, especially super bacteria. Cytotoxicity tests indicated that KGM/Gelatin@Au NPs/GS showed basically no cytotoxicity to L929 cells. In addition, KGM/Gelatin@Au NPs/GS possesses good water absorption, water retention, and enhanced mechanical properties, which can provide a moist environment for wounds and promote healing. In conclusion, our study showed that the antibacterial activity of KGM/Gelatin@Au NPs/GS is better than that of only GS and that it efficiently eliminated super bacteria. Therefore, KGM/Gelatin@Au NPs/GS can be used for killing superbugs, inhibiting bacterial growth, and promoting wound healing.

Silver nanoparticles in situ synthesized by polysaccharides from Sanghuangporus sanghuang and composites with chitosan to prepare scaffolds for the regeneration of infected full-thickness skin defects.

In recent years, silver nanoparticles have widely been used in antibacterial dressings to solve antibiotic resistance problems. However, traditional methods for reducing silver nanoparticles are usually toxic. To overcome this problem, Sanghuangporus sanghuang polysaccharides (FSHPs) were used as a green reducing agent to prepare silver nanoparticles (AgNPs) with a size of 3-35 nm. The FSHPs­silver nanoparticles (FSHPs-Ag) composite with chitosan solution were then freeze-dried to obtain a porous sponge dressing of chitosan-FSHPs-Ag (CS-FSHPs-Ag). The internal pores of CS-FSHPs-Ag were between 50 and 100 µm and had good swelling and water retention properties, which could provide a moist environment for wounds. Based on the experimental results, the appropriate concentration of AgNPs required for CS-FSHPs-Ag to inhibit Escherichia coli and Staphylococcus aureus was determined. Moreover, there was no statistically significant difference between the material treatment and the blank control group, indicating that the material almost showed no toxicity to L929 cells. Finally, this material was used for dressing animal wounds. The results showed that the CS-FSHPs-Ag promoted wound contraction and internal tissue growth better than the wounds treated with Aquacel® Ag, which indicated that the CS-FSHPs-Ag has a great potential as an ideal wound dressing material.

In situ reduction of silver nanoparticles by chitosan-l-glutamic acid/hyaluronic acid: Enhancing antimicrobial and wound-healing activity.

Spongy composites with silver nanoparticles (AgNPs) were synthesized by freeze-drying a mixture of silver nitrate (AgNO3) and chitosan-l-glutamic acid (CG) derivative loaded with hyaluronic acid (HA) solution. CG/AgNP spongy composites had an interconnected porous structure and rough surfaces. When AgNPs (5-20nm) were immobilized on these spongy composites, AgNP aggregation was dependent on AgNO3 concentration. The spongy composites exhibited good mechanical properties, swelling, and water retention capacity. In vitro antibacterial activity showed that the CG/AgNP spongy composites effectively inhibited bacterial (Escherichia coli and Staphylococcus aureus) growth and penetration. Spongy composites containing low concentrations of AgNP were non-toxic to L929 cells, while CG/HA/AgNP spongy composites promoted wound healing, as determined by in vivo tests, wound contraction ratio, average healing time, and histological examination. These results indicate that the spongy composites can serve as effective antibacterial wound dressings.

Accelerated wound-healing capabilities of a dressing fabricated from silkworm cocoon.

Silk fibroin materials have shown some success in wound dressing applications; however, their use for this purpose remains limited by a complex production process and wasted sericin. In the present study, Bombyx mori cocoon materials are used because the protective function of the silkworm cocoon resembles the manner in which the skin protects the human body. A series of silkworm cocoon sol-gel film (SCSF) wound dressings are prepared by immersion in a CaCl2-ethanol-H2O solution for different treatment times. The accelerated wound-healing capabilities of SCSFs are systematically evaluated. Among them, the SCSF sample immersed for 90min exhibits stronger biocompatibility and antibacterial performance compared to other SCSFs. SCSF-90 also exhibits excellent transparency, a high swelling ratio, and good extensibility. Furthermore, in vivo experiments indicate that SCSF-90 can significantly accelerate the healing rate of wounds in New Zealand white rabbits, compared to the standard Mepitel® dressing, and histological examinations reveal that SCSF-90 aided in the successful reconstruction of intact and thickened epidermis. These results demonstrate that the proposed approach may be utilized in the design of antibacterial materials with promising applications in wound dressing.