The antitumor effects of herbal medicine Triphala on oral cancer by inactivating PI3K/Akt signaling pathway: based on the network pharmacology, molecular docking, in vitro and in vivo experimental validation.

BACKGROUND: This research aimed to investigate the anti-tumor effects and underlying genetic mechanisms of herbal medicine Triphala (TRP) in oral squamous cell carcinoma (OSCC). METHODS: The target genes of Triphala (TRP) in oral squamous cell carcinoma (OSCC) were identified, and subsequent functional enrichment analysis was conducted to determine the enriched signaling pathways. Based on these genes, a protein-protein interaction network was constructed to identify the top 10 genes with the highest degree. Genes deregulated in OSCC tumor samples were identified to be hub genes among the top 10 genes. In vitro experiments were performed to investigate the influence of TRP extracts on the cell metabolic activity, migration, invasion, apoptosis, and proliferation of two OSCC cell lines (CAL-27 and SCC-9). The functional rescue assay was conducted to investigate the effect of applying the inhibitor and activator of an enriched pathway on the phenotypes of cancer cells. In addition, the zebrafish xenograft tumor model was established to investigate the influence of TRP extracts on tumor growth and metastasis in vivo. RESULTS: The target genes of TRP in OSCC were prominently enriched in the PI3K-Akt signaling pathway, with the identification of five hub genes (JUN, EGFR, ESR1, RELA, and AKT1). TRP extracts significantly inhibited cell metabolic activity, migration, invasion, and proliferation and promoted cell apoptosis in OSCC cells. Notably, the application of TRP extracts exhibited the capacity to downregulate mRNA and phosphorylated protein levels of AKT1 and ESR1, while concomitantly inducing upregulation of mRNA and phosphorylated protein levels in the remaining three hub genes (EGFR, JUN, and RELA). The functional rescue assay demonstrated that the co-administration of TRP and the PI3K activator 740Y-P effectively reversed the impact of TRP on the phenotypes of OSCC cells. Conversely, the combination of TRP and the PI3K inhibitor LY294002 further enhanced the effect of TRP on the phenotypes of OSCC cells. Remarkably, treatment with TRP in zebrafish xenograft models demonstrated a significant reduction in both tumor growth and metastatic spread. CONCLUSIONS: Triphala exerted significant inhibitory effects on cell metabolic activity, migration, invasion, and proliferation in OSCC cell lines, accompanied by the induction of apoptosis, which was mediated through the inactivation of the PI3K/Akt pathway.

Controllable Adaptive Molybdate-Oligosaccharide Nanoparticles Regulate M2 Macrophage Mitochondrial Function and Promote Angiogenesis via PI3K/HIF-1α/VEGF Pathway to Accelerate Diabetic Wound Healing.

The complex wound environment of diabetic wounds leads to poor treatment efficacy, and the inflammatory disorders and vascular injury are the primary causes of death in such patients. Herein, a sprayable, controllable adaptive, pH-responsive nanosystem of molybdate and oligosaccharide (CMO) is specially developed as an immunomodulatory and angiogenesis-promotion material for diabetic wound healing. CMO exhibited pH-responsive release of Mo2+ and oligosaccharide (COS), specifically in response to the alkalescent environment observed in diabetic wounds. CMO provide an anti-inflammatory environment by promoting M2 polarization through significantly stimulating macrophage mitochondrial function. Specifically, CMO with a certain concentration reduce reactive oxygen species (ROS) and tumor necrosis factor α (TNF-α) expression, and upregulated mitochondrial membrane potential (MMP), superoxide dismutase (SOD), and interleukin 10 (IL-10) expression in macrophages. Moreover, CMO facilitate angiogenesis via upregulating the PI3K/HIF-1α/VEGF pathway-a critical process for the formation of new blood vessels that supply nutrients and oxygen to the healing tissue. Remarkably, CMO promote cell viability and migration of endothelial cells, and enhance the expression of angiogenic genes. In vitro and in vivo studies suggest this simple but powerful nanosystem targeting mitochondrial function has the potential to become an effective treatment for diabetic wound healing.

SERPINB5 is a novel serum diagnostic biomarker for gastric high-grade intraepithelial neoplasia and plays a role in regulation of macrophage phenotypes.

BACKGROUND: Gastric cancer (GC) develops from gastric precancerous lesions (GPL), and early diagnosis and treatment at the premalignant stage may achieve a higher benefit‒cost ratio with a reduced necessity for surgery. However, reliable noninvasive screening biomarkers of GPL are currently lacking. METHODS: The marker genes of GPL encoding extracellular proteins were identified by bioinformatics analysis and further verified by immunofluorescence and immunohistochemistry assays. Serum samples were collected to measure the levels of SERPINB5, the diagnostic efficacy of which was assessed by the area under the receiver operating characteristic (ROC) curve (AUC). Finally, the effect of SERPINB5 on the phenotypic conversion of macrophages was verified by public data and in vitro experiments. RESULTS: SERPINB5 was identified as an extracellular biomarker of GPL that had good diagnostic efficacy. High expression of SERPINB5 was observed in the epithelial cells and adjacent extracellular matrix on sections of gastric high-grade intraepithelial neoplasia (HGIN). Importantly, SERPINB5 determined in serum was significantly increased in the HGIN group, and the AUC for discriminating between HGIN and chronic gastritis or low-grade intraepithelial neoplasia was 0.9936 and 0.9750, respectively. Moreover, SERPINB5 expression was positively correlated with macrophage infiltration, and M1 marker NOS2 expression, but negatively correlated with M2 marker CSF1R expression. In THP-1-derived macrophages, SERPINB5 upregulated expression of M1-related cytokines TNF-α and IL-12, and M1 marker CD86, but suppressed production of M2-related cytokines TGF-ß and IL-10. CONCLUSIONS: Our study provides evidence that SERPINB5 may serve as a promising noninvasive serum biomarker for gastric HGIN screening and regulate macrophage phenotype conversion.

In vitro and in vivo antimicrobial activity of antimicrobial peptide Jelleine-I against foodborne pathogen Listeria monocytogenes.

As a human foodborne pathogen, Listeria monocytogenes can cause severe human listeriosis and develop resistance to antibiotics. Antimicrobial peptides (AMPs) are produced from all kingdoms of life and regarded as promising alternatives to conventional antibiotics. Jelleine-I is an AMP identified from honeybees royal jelly. In this study, we explored the activity and action mechanism of Jelleine-I against L. monocytogenes. We found its minimum inhibitory concentration to be 12.5 µg/mL. Membrane permeability analysis revealed that Jelleine-I increased L. monocytogenes cell membrane permeability, causing calcium leakage. Scanning, transmission electron microscopy and fluorescence microscopy revealed that Jelleine-I destroyed membrane integrity, disrupted intracellular structures and interacted with the bacterial DNA. DNA binding analysis demonstrated that Jelleine-I bound to bacterial genomic DNA. Results of reverse transcription-quantitative PCR revealed that Jelleine-I affected bacterial DNA replication gene expression levels. Moreover, Jelleine-I induced cellular reactive oxygen species (ROS) production from fluorescence intensity analysis, and inhibited bacterial biofilm formation. Results of immunomodulation in Galleria mellonella revealed that Jelleine-I increased host hemocyte counts, upregulated host AMP gene (Gloverin and Cecropin D) expression, and inhibited proinfammatory cytokine (tumor necrosis factor α and interleukin 6) production induced by bacterial infection. It efficiently killed bacteria and increased the survival rate of infected insects to 70 %. Furthermore, Jelleine-I increased the G1 to S phase transition in mammalian cells from cells cycle analysis, and cytotoxicity assay results indicated that it promoted cell proliferation without hemolysis or cytotoxicity. Collectively, Jelleine-I possesses antimicrobial, immunomodulatory and cell proliferative activities, and is a promising candidate for preventing L. monocytogenes emergence and dissemination.

Impacts of chitosan oligosaccharide (COS) on angiogenic activities.

It has been proved that chitosan oligosaccharide (COS) has a more favorable therapeutic applications such as wound healing and anti-tumor treatment, and can affect angiogenesis. For better understanding the effect of COS on angiogenic activities at cellular level, COS with different concentration and degree of polymerization (DP) were used to culture human umbilical vein endothelial cells (HUVECs) in this work. Cell proliferation activity, cell morphology, cell migration and angiogenesis associated factor expression of HUVECs were evaluated. The results indicated that COS at a high concentration of 400 µg/mL (COS(400)) and DP of 6 (Chitinhexaose Hydrochloride, COS6) had inhibitory effect on angiogenic activities of HUVECs. Specifically, COS(400) and COS6 inhibited cell proliferation activity, cell migration, and vascular endothelial cell growth factor (VEGF) expression of HUVECs. While COS at a low concentration (<400 µg/mL) and suitable polymerization degrees (DP < 6) had little significant effect on cell proliferation, migration, and VEGF expression of HUVECs, showing dose-dependent effect. These findings provided insight for the potential use of COS, for broadening its future applications in biomedical fields and functional materials area. It also helped guide the design and synthesis of chitosan-based materials as an angiogenesis inhibitor for anti-angiogenic therapy.

Macrophage Polarization Mediated by Chitooligosaccharide (COS) and Associated Osteogenic and Angiogenic Activities.

The host response to implanted biomaterials can influence the functionality of the materials and modulate the tissue repair and remolding. Macrophages, key cells in the host response to biomaterials, can be polarized into different phenotypes, which are important in regenerative medicine. The objective of this study was to evaluate the effect of chitooligosaccharide (COS) on the modulation of macrophage (RAW 264.7) polarization and the associated osteogenic and angiogenic activities. The results demonstrate that COS can shift the macrophage response to an alternatively activated reparative response, which can then upregulate the expression of anti-inflammatory cytokines. COS can also create an immune-modulated microenvironment, with osteogenesis- and angiogenesis-related proteins and a biological process that further influences the osteogenic/angiogenic differentiation and promotion of bone mesenchymal stem cells (BMSCs) and vascular activation of human umbilical vein endothelial cells (HUVECs). In this work, at a low concentration of 4 µg/mL [COS(4)] and suitable polymerization degree of 5 (chitopentaose hydrochloride, COS5) of COS, the associated effect on an alternatively activated reparative response and upregulation of anti-inflammatory cytokine expression was better than that of COS at other concentrations or polymerization degrees. The supernatant from a culture of RAW 264.7 stimulated by COS(4) and COS5 [conditioned medium S-COS(4) and S-COS5] contained more osteogenesis- and angiogenesis-related proteins like DKK-1, OPN, osteoactivin, vascular endothelial growth factor (VEGF) R1, epidermal growth factor (EGF), and insulin-like growth factor binding protein-5 (IGFBP-5) for regulation of osteogenesis/angiogenesis. Specifically, the alkaline phosphatase (ALP) activity and typical osteogenesis-related proteins of BMSCs were significantly influenced by the conditioned media of COS-stimulated macrophages [S-COS(4) and S-COS5]. Furthermore, the conditioned media affected HUVEC proliferation and migration for vascularization. Our results suggest that COS at a low concentration and suitable polymerization degrees has a beneficial effect on immunity modulation (an alternatively activated reparative response) and can modulate osteogenesis/angiogenesis processes for tissue regeneration without using any inductive agent.

[Feasibility of multi models targeting home pain alleviation service used on 220 cases with advanced cancer].

OBJECTIVE: To investigate the feasibility and effectiveness of multiple modes on home pain alleviation service used for advanced cancer patients to in prove clinical therapy services. METHODS: The study was involved with 220 patients with advanced cancers to provide them with multimodal analgesia services at home, from February 2010 to February 2013. Patients in this study had been taking both opioid treatments. They were randomly divided into two groups with the number as 112 and 108 and were given different doses of morphine or other drugs. During the period of observation, data was collected under the M. D. Anderson symptom Inventory (MDASI) score and classification of score on pain. RESULTS: Differences of pain scores in the two groups and the MDASI score were significant and presented as skewed distribution. Scores on pain score were between groups were significantly different (Z = -9.735, P < 0.001). The average rankings of A group and B group were 76.68 and 162.79 respectively. Under the application of 0.4 mg alprazolam, the degree of pain alleviation seemed to be better. The differences on comprehensive scores between different drug groups were statistically significant (Z = -13.334, P < 0.001). The average rankings of groups A and B were respectively 59.87 and 179.08. Under the use of 0.4 mg of alprazolam, the results could be considered to show better improvements in symptomatic patients. Application of 0.4 mg alprazolam on patients with advanced-cancer-induced-pain showed a better symptomatic improvement than using morphine. CONCLUSION: Patients with advanced cancers receiving multimodal analgesia short-term sedation therapies at home, showed both ideal feasibility and good effectiveness. When morphine was combined used with midazolam at home, a better outcome could be seen in pain-releasing on patients with cancer, than single morphine analgesia was used.