Polysaccharides as antioxidants and prooxidants in managing the double-edged sword of reactive oxygen species.

Polysaccharides, a class of naturally occurring carbohydrates, were widely presented in animals, plants, and microorganisms. Recently, health benefits of polysaccharides have attracted much attention due to their unique characteristics in reactive oxygen species (ROS) management. ROS, by-products of aerobic metabolism linked to food consumption, exhibited a dual role in protecting cells and fostering pathogenesis collectively termed double-edged sword. Some interesting studies reported that polysaccharides could behave as prooxidants under certain conditions, besides antioxidant capacities. Potentiation of the bright side of ROS could contribute to the host defense that was vitally important for the polysaccharides acting as biological response modifiers. Correspondingly, disease prevention of polysaccharides linked to the management of ROS production was systematically described and discussed in this review. Furthermore, major challenges and future prospects were presented, aiming to provide new insight into applying polysaccharides as functional food ingredients and medicine.

Effect of a dynamic navigation device on the accuracy of implant placement in the completely edentulous mandible: An in vitro study.

STATEMENT OF PROBLEM: A less invasive and more convenient workflow is needed for dynamic navigation-guided implant surgery for the edentulous arch. PURPOSE: The purpose of this in vitro study was to evaluate the accuracy of a novel dynamic navigation device developed for the completely edentulous mandible. MATERIAL AND METHODS: Two temporary 1-piece mini-implants were placed in the anterior region of a completely edentulous mandibular model for fixation of the navigation device. A total of 40 implants were inserted in 10 completely edentulous mandibular models with the aid of the dynamic navigation device. The accuracy of placement was determined by overlapping the preoperative plan with the postoperative cone beam computed tomography (CBCT) scans. The difference in the accuracy at different implant positions was compared by MANOVA and Bonferroni-corrected ANOVAs. The difference in accuracy between implants #1-20 and #21-40 was assessed for learning progression. RESULTS: The deviation of the implants (mean ±standard deviation) was 1.14 ±0.50 mm at the entry point and 1.29 ±0.48 mm at the apex. The mean ±standard deviation angular deviation was 3.02 ±1.32 degrees. No significant difference was seen between the planned and the placed deviation among the 4 implant positions. After repeated placement with this dynamic approach, implant accuracy at the entry (P=.001) and apex (P=.017) improved significantly. CONCLUSIONS: The navigation device achieved acceptable implant placement accuracy in the edentulous mandible. Deviations between the planned and placed locations were not affected by different implant positions. After repeated operations with this dynamic approach, accuracy at the entry and apex improved significantly.

[Effect of WRAP53 ß Targeted Co-Inhibitory Pathways Based on Comprehensive Bioinformatics Analysis in Treating Squamous Cell Carcinoma of the Head and Neck].

Objective: To investigate the association between WD40-encoding RNA antisense to p53 ( WRAP53 ß), a telomerase new core subunit, and the clinical, genomic and immune infiltration characteristics of squamous cell carcinoma of the head and neck (HNSC), and to explore for potential joint targeted therapy of HNSC. Methods: Tumor IMmune Estimation Resource (TIMER) online modules were adopted to predict the association between WRAP53 ß expression and the clinical features, oncogene, and immune infiltration of HNSC in the Cancer Genome Atlas (TCGA) cohort. Tumor Immune Single-cell Hub (TISCH) was used to analyze WRAP53 ß expression at the single cell level. Analysis of the small molecule inhibitors potentially targeting WRAP53 ß was carried out by Computational Analysis of REsistance (CARE). In the in vitro verification experiment, recombinant lentiviral particles with the sh WRAP53 ß sequence were synthesized. Then, the oral squamous cell carcinoma cell line Cal27 (the sh WRAP53 ßgroup) stably expressing sh WRAP53 ß were constructed, and two control groups were set up (the shNC group consisting of Cal27 cells added with lentiviral particles containing non-specific control sequences and the Con group consisting of untreated Cal27 cells). MTT assay was done to examine the proliferation of cells in the three groups. Cellular immunofluorescence assay was done for further qualitative examination of the expression of P53 protein in the cells of the sh WRAP53 ß group and the shNC group. Western blot was done to measure the expression of WRAP53ß and γ-H2AX, a DNA damage protein, in the 18 th, 23 rd and 28 th passages of the sh WRAP53 ß group and the shNC group. Finally, specimens of 13 cases of oral squamous cell carcinoma and 7 cases of oral mucosal inflammation were collected, and the expression of WRAP53ß and γ-H2AX in the clinical specimens of oral squamous cell carcinoma was verified with immunohistochemistry. Resluts: TIMER analysis revealed that the expression level of WRAP53 ß in HNSC tissues was significantly higher than that in normal tissues. There was a significant positive correlation between WRAP53 ß expression and multiple genes in the p53 pathway, including CCNB1, CCNB2 and CDK1. Although no significant correlation between WRAP53 ß expression and infiltrating immune cells was found, WRAP53 ß was significantly positively correlated with the inflammatory factors IFN-γ and IL23A, and negatively correlated with IL-1A and IL-6 in HPV-positive carcinoma of the head and neck. TISCH single cell sequencing datasets also showed higher expression of WRAP53 ß in malignant cells, and very low or zero expression in immune cells. According to the CARE scores, the most potent WRAP53 ß co-inhibitory drugs were ATM, CDK1 and MDM4 targeted inhibitors. In vitro cell experiments showed that the proliferation ability of Cal27 cells decreased significantly in the sh WRAP53 ß group as compared with that of the control group between Day 5 and Day 7 ( P<0.05). Furthermore, the expression of P53 decreased significantly in the sh WRAP53 ß group. As compared with the control group, the expression of WRAP53ß in sh WRAP53 ß group significantly decreased in the 18 th, 23 rd and 28 th passages ( P<0.05), while γ-H2AX expression only decreased in the 18 th and 28 th passages ( P<0.05) according to the results of Western blot. Clinical specimens showed rather high positive expression rate of γ-H2AX in oral squamous cell carcinoma tissues (12/13), while the expression of WRAP53ß was not detected in oral mucositis samples (0/7). Conclusions: WRAP53 ß showed significantly higher expression level in HSNC, and was significantly associated with p53 pathway genes. ATM, CDK1 and MDM4 inhibitors may be potential WRAP53 ß co-inhibitory agents. RNA interference of WRAP53 ß expression may cause inhibition of DNA damage, thereby indicating therapeutic potential for HNSC.

Ganoderma atrum polysaccharide modulates the M1/M2 polarization of macrophages linked to the Notch signaling pathway.

Macrophages could be polarized into two major sub-populations including classically activated (M1) and alternatively activated (M2) macrophages. The present study aimed to investigate the effects of Ganoderma atrum polysaccharide (PSG-1) on the regulation of macrophage polarization and further explored the associated molecular mechanisms. In this work, a lipopolysaccharide (LPS) plus IFN-γ and IL-4 were used to establish an in vitro model of two extreme states, namely pro-inflammatory M1 and anti-inflammatory M2. The results showed that PSG-1 had effects on the behavior modification of macrophage polarization by reducing CD80 expression in LPS plus IFN-γ-induced M1 macrophages, and attenuating CD23 expression in IL-4-induced M2 macrophages. Further study revealed that PSG-1-modulated M1 and M2 macrophage polarization was associated with controlling phagocytosis, reactive oxygen species generation, NO and cytokines (IL-1ß, IL-6 and IL-10). Subsequently, the treatment of M1 macrophages with a combination of PSG-1 and a Notch-response inhibitor (DAPT) did not alter CD80 expression compared with DAPT alone, while several pro-inflammatory parameters were considerably decreased, suggesting that the Notch signaling pathway partly mediated the effects of PSG-1 on modulating macrophage polarization. Together, our findings suggested that PSG-1 could repair the chaos in the polarization of M1/M2 macrophages and the molecular mechanism linked to the Notch signaling pathway.

Implant placement in the pterygoid region with dynamically navigated surgery: A clinical report.

Implant placement in the pterygoid region is a reliable treatment for posterior maxillary tooth loss. However, the surgery is not widely applied because the implant placement region is hard to access and the direct visual access is limited. This clinical report describes the use of a dynamic navigation system to improve the pterygoid implant placement surgery. Real-time imaging of the surgery area and full-time guidance are provided by the system to alleviate the problem of lack of visibility and to reduce the complexity of placement.

Periodontal Pathogens Promote Oral Squamous Cell Carcinoma by Regulating ATR and NLRP3 Inflammasome.

Periodontitis is closely related to oral cancer, but the molecular mechanism of periodontal pathogens involved in the occurrence and development of oral cancer is still inconclusive. Here, we demonstrate that, in vitro, the cell proliferation ability and S phase cells of the periodontitis group (colonized by Porphyromonas gingivalis and Fusobacterium nucleatum, P+) significantly increased, but the G1 cells were obviously reduced. The animal models with an in situ oral squamous cell carcinoma (OSCC) and periodontitis-associated bacteria treatment were constructed, and micro-CT showed that the alveolar bone resorption of mice in the P+ group (75.3 ± 4.0 µm) increased by about 53% compared with that in the control group (48.8 ± 1.3 µm). The tumor mass and tumor growth rate in the P+ group were all higher than those in the blank control group. Hematoxylin-eosin (H&E) staining of isolated tumor tissues showed that large-scale flaky necrosis was found in the tumor tissue of the P+ group, with lots of damaged vascular profile and cell debris. Immunohistochemistry (IHC) of isolated tumor tissues showed that the expression of Ki67 and the positive rate of cyclin D1 were significantly higher in tumor tissues of the P+ group. The qRT-PCR results of the expression of inflammatory cytokines in oral cancer showed that periodontitis-associated bacteria significantly upregulated interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-18, apoptosis-associated speck-like protein containing a CARD (ASC) (up to six times), and caspase-1 (up to four times), but it downregulated nuclear factor (NF)-κB, NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3), and IL-1ß (less than 0.5 times). In addition, the volume of spleen tissue and the number of CD4+ T cells, CD8+ T cells, and CD206+ macrophages in the P+ group increased significantly. IHC and Western blotting in tumor tissues showed that expression levels of γ-H2AX, p-ATR, RPA32, CHK1, and RAD51 were upregulated, and the phosphorylation level of CHK1 (p-chk1) was downregulated. Together, we identify that the periodontitis-related bacteria could promote tumor growth and proliferation, initiate the overexpressed NLRP3, and activate upstream signal molecules of ATR-CHK1. It is expected to develop a new molecular mechanism between periodontitis-related bacteria and OSCC.

MPI-based bioinformatic analysis and co-inhibitory therapy with mannose for oral squamous cell carcinoma.

Mannose induces tumor cell apoptosis and inhibits glucose metabolism by accumulating intracellularly as mannose 6-phosphate while the drug sensitivity of tumors is negatively correlated with mannose phosphate isomerase gene (MPI) expression. In this study, we performed a first attempt to explore the relationship between the targeted gene MPI and immune infiltration and genetic and clinical characteristics of head and neck squamous carcinoma (HNSC) using computational algorithms and bioinformatic analysis, and further to verify the co-inhibition effects of mannose with genotoxicity, immune responses, and microbes dysbiosis in oral squamous cell carcinoma (OSCC) in vitro and in vivo. Our results found that patients with lower MPI expression had higher survival rate. The enhancement of MPI expression was in response to DNA damage gene, and ATM inhibitor was verified as a potential drug with a synergistic effect with mannose on HSC-3. In the HNSC, infiltrated immunocytes CD8+ T cell and B cell were the significantly reduced risk cells, while IL-22 and IFN-γ showed negative correlation with MPI. Finally, mannose could reverse immunophenotyping caused by antibiotics in mice, resulting in the decrease of CD8+ T cells and increase of myeloid-derived suppressor cells (MDSCs). In conclusion, the MPI gene showed a significant correlation with immune infiltration and genetic and clinical characteristics of HNSC. The treatment of ATM inhibitor, immune regulating cells of CD8+ T cells and MDSCs, and oral microbiomes in combination with mannose could exhibit co-inhibitory therapeutic effect for OSCC.

Chimonanthus nitens Oliv. essential oil mitigates lipopolysaccharide-induced acute lung injury in rats.

This study aimed to evaluate the effect of Chimonanthus nitens Oliv. essential oil (named CEO) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats. In the present study, 21 compounds were characterized in CEO by gas chromatography-mass spectrometry analysis. Furthermore, animal data suggested that CEO could protect rats against ALI, as evidence by increasing white blood cell count, reducing immune organ index and improving lung histopathological changes in rats subjected to LPS. Reduction of the levels of IL-1ß was also shown during CEO-triggering lung protection in rats. Meanwhile, these protective effects of CEO were accompanied by the attenuation of lipid oxidation, and elevation of antioxidant enzymes, suggesting that enhancement of antioxidant defense was linked to its lung protection. Moreover, a combination with CEO and LPS significantly elevated short-chain fatty acids (SCFAs) compared with LPS alone via increasing propionic, i-butyric, butyric and i-valeric acid on LPS-induced ALI in rats. Therefore, our findings indicated that CEO could alleviate LPS-caused ALI in rats by controlling aberrant inflammation, correcting the redox system, and modulating SCFAs in rats.

Mannose Receptor Mediates the Immune Response to Ganoderma atrum Polysaccharides in Macrophages.

The ability of mannose receptor (MR) to recognize the carbohydrate structures is well-established. Here, we reported that MR was crucial for the immune response to a Ganoderma atrum polysaccharide (PSG-1), as evidenced by elevation of MR in association with increase of phagocytosis and concentrations of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) in normal macrophages. Elevation of MR triggered by PSG-1 also led to control lipopolysaccharide (LPS)-triggered inflammatory response via the increase of interleukin-10 (IL-10) and inhibition of phagocytosis and IL-1ß. Anti-MR antibody partly attenuated PSG-1-mediated anti-inflammatory responses, while it could not affect TNF-α secretion, suggesting that another receptor was involved in PSG-1-triggered immunomodulatory effects. MR and toll-like receptor (TLR)4 coordinated the influences on the TLR4-mediated signaling cascade by the nuclear factor-κB (NF-κB) pathway in LPS-stimulated macrophages subjected to PSG-1. Collectively, immune response to PSG-1 required recognition by MR in macrophages. The NF-κB pathway served as a central role for the coordination of MR and TLR4 to elicit immune response to PSG-1.

(-)-Epigallocatechin-3-gallate alleviates doxorubicin-induced cardiotoxicity in sarcoma 180 tumor-bearing mice.

AIMS: (-)-Epigallocatechin-3-gallate (EGCG), a major green tea polyphenol compound, plays an important role in the prevention of cardiovascular disease and cancer. The present study aimed to investigate the effects of EGCG on doxorubicin (DOX)-induced cardiotoxicity in Sarcoma 180 (S180) tumor-bearing mice. MAIN METHODS: S180 tumor-bearing mice were established by subcutaneous inoculation of S180 cells attached to the axillary region. The extent of myocardial injury was accessed by the amount of lactate dehydrogenase (LDH) released in serum. Heart tissue was morphologically studied with transmission electron microscopy. Apoptosis, reactive oxygen species (ROS) generation, mitochondrial membrane potential (ΔÑ°m) as well as calcium concentration were measured by flow cytometric analysis. Expression levels of manganese superoxide dismutase (MnSOD) were analyzed by Western blot. KEY FINDINGS: Results showed that the combination with EGCG and DOX significantly inhibited tumor growth and enhanced induction of apoptosis compared with DOX alone. Moreover, administration of EGCG could suppress DOX-induced cardiotoxicity as evidenced by alleviating LDH release and apoptosis in cardiomyocyte. EGCG-evoked cardioprotection was in association with the increase of ΔÑ°m and MnSOD expression. EGCG was also found to attenuate ROS generation and myocardial calcium overload in Sarcoma 180 tumor-bearing mice subjected to DOX. SIGNIFICANCE: EGCG alleviated DOX-induced cardiotoxicity possibly in part mediated by increasing of MnSOD and Ñ°m, reducing myocardial calcium overload and subsequently attenuating the apoptosis and LDH release. Our findings suggest that co-administration of EGCG and DOX have potential as a feasible strategy to mitigate cardiotoxicity of DOX without compromising its chemotherapeutic value.

Ganoderma atrum polysaccharide ameliorates ROS generation and apoptosis in spleen and thymus of immunosuppressed mice.

Ganoderma atrum polysaccharide (PSG-1) is a bioactive compound with antioxidant and immunomodulatory activities. The aim of this study was to determine the effect of PSG-1 on reactive oxygen species (ROS) generation and apoptosis in spleen and thymus of cyclophosphamide (CTX)-induced immunosuppressed mice. The results showed that PSG-1 protected mice against CTX-mediated immunosuppression, as evidenced by enhancing the ratios of thymus and spleen weights to body weight, promoting T cell and B cell survival, and increasing levels of TNF-α and IL-2. Apoptosis, ROS generation and lipid peroxidation in the immune organs of the immunosuppressed animals were ameliorated by PSG-1. The immune benefits of PSG-1 were associated with the enhancement of the activities of glutathione peroxidase, superoxide dismutase and catalase in the immune organs, implying that antioxidant activities of PSG-1 may play an important role in PSG-1-evoked immune protection. Taken together, these findings have demonstrated that PSG-1 may ameliorate CTX-induced immunosuppression through reducing apoptosis and oxidative damage in immunological system.

[Analysis of causes and whole microbial structure in a case of rampant caries].

OBJECTIVE: To analyze the whole microbial structure in a case of rampant caries to provide evidence for its prevention and treatment. METHODS: Clinical samples including blood, supragingival plaque, plaque in the caries cavity, saliva, and mucosal swabs were collected with the patient's consent. The blood sample was sent for routine immune test, and the others samples were stained using Gram method and cultured for identifying colonies and 16S rRNA sequencing. DNA was extracted from the samples and tested for the main cariogenic bacterium (Streptococcus mutans) with qPCR, and the whole microbial structure was analyzed using DGGE. RESULTS: The patient had a high levels of IgE and segmented neutrophils in his blood. Streptococci with extremely long chains were found in the saliva samples under microscope. Culture of the samples revealed the highest bacterial concentration in the saliva. The relative content of hemolytic bacterium was detected in the samples, the highest in the caries cavity; C. albicans was the highest in the dental plaque. In addition, 33 bacterial colonies were identified by VITEK system and 16S rDNA sequence phylogenetic analysis, and among them streptococci and Leptotrichia wade were enriched in the dental plaque sample, Streptococcus mutans, Fusobacterium nucleatum, and Streptococcus tigurinus in the caries cavity, and Lactobacillus in the saliva. S. mutans was significantly abundant in the mucosal swabs, saliva and plaque samples of the caries cavity as shown by qPCR. Compared to samples collected from a healthy individual and another two patients with rampant caries, the samples from this case showed a decreased bacterial diversity and increased bacterial abundance shown by PCR-DGGE profiling, and multiple Leptotrichia sp. were detected by gel sequencing. CONCLUSION: The outgrowth of such pathogenic microorganisms as S. mutans and Leptotrichia sp., and dysbiosis of oral microbial community might contribute to the pathogenesis of rampant caries in this case.