Comparison of Easydo Activator, ultrasonic and needle irrigation techniques on sealer penetration and smear layer removal in vitro.

The effects of Easydo Activator (EA), a new sonic irrigation system, on sealer penetration at the root apex were compared to needle irrigation (NI) and passive ultrasonic irrigation (PUI) in this study. Forty-two single-rooted teeth were prepared and randomly divided into three groups (n = 14): group 1: NI; group 2: PUI; and group 3: EA. A solution of 3% sodium hypochlorite (NaOCl) was used for irrigation. Nine teeth in each group were filled with AH Plus sealer mixed with CY5 fluorescent dye and a single gutta-percha cone. The sealer penetration area, maximum penetration depth and percentage of sealer penetration at 5 mm and 1 mm from the apex were analyzed by confocal laser scanning microscopy (CLSM). The remaining 5 teeth in each group were subjected to test smear layer scores by scanning electron microscopy (SEM). The CLSM evaluation showed that increases in the area, depth and percentage of sealer penetration were detected at 1 and 5 mm from the root apex in the PUI group compared with the NI group, and greater increases were observed in the EA group (P < 0.05). The SEM experiment showed that the lowest scores for the smear layer and debris removal were achieved by the EA group when compared with the PUI and NI groups (P < 0.05). In conclusion, EA was superior to PUI and NI regarding sealer penetration at the root apex during endodontic treatment, and it could provide a new technical idea for clinical root canal therapy.

Notch1 is a marker for in situ resting osteocytes in a 3-dimensional gel culture model.

PURPOSE: Osteocytes in vivo exhibit different functional states, but no specific marker to distinguish these is currently available. MATERIALS AND METHODS: To simulate the differentiation process of pre-osteoblasts to osteocytes in vitro, MC3T3-E1 cells were cultured on type I collagen gel and a three-dimensional (3D) culture system was established. The Notch expression of osteocyte-like cells in 3D culture system was compared with that of in situ osteocytes in bone tissues. RESULTS: Immunohistochemistry demonstrated that Notch1 was not detected in "resting" in situ osteocytes, but was detected in normal cultured osteocyte-like cell line MLO-Y4. Osteocytes obtained from conventional osteogenic-induced osteoblasts and long-term cultured MLO-Y4 cells could not replicate the Notch1 expression pattern from in situ osteocytes. From day 14-35 of osteogenic induction, osteoblasts in 3D culture system gradually migrated into the gel to form canaliculus-like structures similar to bone canaliculus. On day 35, stellate-shaped osteocyte-like cells were observed, and expression of DMP1 and SOST, but not Runx2, was detected. Notch1 was not detected by immunohistochemistry, and Notch1 mRNA level was not significantly different from that of in situ osteocytes. In MC3T3-E1 cells, down-regulation of Notch2 increased Notch1, Notch downstream genes (ß-catenin and Nfatc1), and Dmp1. In MLO-Y4 cells, Notch2 decreased after Notch1 siRNA transfection. Downregulation of Notch1 or Notch2 decreased Nfatc1, ß-catenin, and Dmp1, and increased Sost. CONCLUSIONS: We established "resting state" osteocytes using an in vitro 3D model. Notch1 can be a useful marker to help differentiate the functional states of osteocytes (activated vs. resting state).

Comparison of EASYDO ACTIVATOR, passive ultrasonic, and needle irrigation techniques on the treatment of apical periodontitis: a study in rats.

OBJECTIVES: To evaluate the long-term therapeutic effect of EASYDO ACTIVATOR, passive ultrasonic irrigation, and needle irrigation in experimental apical periodontitis in rats. MATERIALS AND METHODS: Sprague-Dawley male rats were used to produce periapical lesions. The pulp chambers of the bilaterally first mandibular molars were exposed and left open for 21 days. The rats were divided into four groups according to different irrigation protocols. Seven days after irrigation, the mandibles were removed for micro-CT, histological, and immunohistochemical analysis. Serum levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were assessed by enzyme-linked immunosorbent assays (ELISA). Statistical data were analyzed by one-way analysis of variance (ANOVA) with LSD tests. RESULTS: The passive ultrasonic irrigation and EASYDO ACTIVATOR groups had the smallest apical lesions compared to the other groups (P < 0.05), while the needle irrigation group had smaller lesions than the control group (P < 0.05). The EASYDO ACTIVATOR group had less inflammation infiltration compared to the control and needle irrigation groups (P < 0.05). The control and needle irrigation groups had more TNF-α expression compared to the passive ultrasonic irrigation and EASYDO ACTIVATOR groups (P < 0.05). The lowest IL-6 expression was observed in the EASYDO ACTIVATOR group. The EASYDO ACTIVATOR group had the lowest serum level of TNF-α than other groups (P < 0.05). IL-6 expression was significantly lower in the EASYDO ACTIVATOR group in comparison with the control and needle irrigation groups (P < 0.05). CONCLUSIONS: EASYDO ACTIVATOR can significantly reduce the apical lesions and decrease the inflammatory response around the periapical area. CLINICAL RELEVANCE: EASYDO ACTIVATOR is recommended for clinical application.

Role of NOD2 and hepcidin in inflammatory periapical periodontitis.

The immunological response occurring during periapical inflammation includes expression of nucleotide binding oligomerization domain containing 2 and hepcidin. Nucleotide binding oligomerization domain containing 2 deficiency increases infiltration of inflammatory cells close to alveolar bone. Hepcidin has an important role in iron metabolism affecting bone metabolism.We investigated the role of nucleotide binding oligomerization domain containing 2 and hepcidin in inflammatory periapical periodontitis. Periapical periodontitis was induced in rats and confirmed by micro-computed tomography. Nucleotide binding oligomerization domain 2 and hepcidin were evaluated through immunohistochemistry. Bioinformatics analysis was undertaken usingthe Kyoto Encyclopedia of Genes and Genomes and Gene Ontology databases. Micro-computer tomography revealed alveolar bone resorption in the periapical region and furcation area of mandibular molars in rats of the periapical periodontitis group. Immunohistochemistry showed increased expressionof nucleotide binding oligomerization domain containing 2 and hepcidin around root apices in rats of the periapical periodontitis group. Bioinformatics analysis of differentially expressed genes in inflamed and non-inflamed tissues revealed enrichment in the NOD-like receptor signaling pathway. Our data suggest that nucleotide binding oligomization domain contain2 and hepcidin have important roles in periapical periodontitis severity because they can reduce alveolar bone loss.They could elicit new perspectives for development of novel strategies for periapical periodontitis treatment.

Comparison of sealer penetration of sonic activation versus conventional needle irrigation: a systematic review and meta-analysis of randomized controlled trials.

BACKGROUND: Most existing studies comparing the efficiency of sonic irrigation (SI) and conventional needle irrigation (CNI) in increasing the penetration of sealers into dentine tubules are controversial; and this study aimed to determine whether the use of SI can lead to greater sealing ability than CNI, during the root canal treatment. METHODS: The EMBASE, PubMed, and Cochrane Library databases were used to find confocal laser scanning microscopy studies evaluating percentage and maximum depth of sealer penetration following the use of SI or CNI in mature permanent teeth until October 2022. The critical estimative checklist of randomized controlled trials of the standardized Joanna Briggs Institute was adopted to independently score the quality of each study. The random-effect model for meta-analysis was used to analyse for each canal segment (apical, middle, coronal). The results are shown in the forest plots as weighted mean differences (WMDs) with 95% confidence intervals (95% CIs). RESULTS: Ninety-seven articles were included in the preliminary screening, and nine of them were included in this study. Eight studies were included in the meta-analysis.The meta-analysis exhibited great increases in the coronal (WMD: 8.09, 95% CI 2.78-13.40/WMD: 165.32, 95% CI 128.85-201.80), and middle segments (WMD: 8.81, 95% CI 5.76-11.87/WMD: 132.98, 95% CI 68.71-197.25) for the percentage and maximum depth of sealer penetration, respectively. The percentage of sealer penetration in the apical thirds region was nonsignificant (WMD: 4.73, 95% CI - 2.34-11.80). However, the maximum depth of sealer penetration in the apical thirds region was significant (WMD: 121.46, 95% CI 86.55-156.38). Chi-squared analysis revealed heterogeneity scores of 0.0-70.0% and 44.0-90.0% for the percentage and maximum depth of sealer penetration, respectively. DISCUSSION: This review verified that SI significantly improves tubular dentin sealer penetration in most areas of the root canal; thus, SI may lead to better filling efficiency and anti-reinfection effects than CNI during and after the root canal therapy. Nevertheless, a large heterogeneity in the current data comparing the irrigation efficiency of SI versus CNI in the apical third of the root canal was found, implying the necessity to standardize root canal irrigation procedures and obtain more accurate results in this area. TRIAL REGISTRATION: INPLASY database (INPLASY202270116).

miR-124-3p increases in high glucose induced osteocyte-derived exosomes and regulates galectin-3 expression: A possible mechanism in bone remodeling alteration in diabetic periodontitis.

The mechanisms underlying the two-way relationship between diabetes mellitus (DM) and periodontitis are unclear. We examined a possible effect of galectin-3 (Gal-3), a factor in DM and bone metabolism, on periodontitis with or without DM. Using enzyme-linked immunosorbent assay, we detected saliva Gal-3 in patients with periodontitis, with or without type 2 diabetes mellitus (T2DM). In animal models, we measured periodontal bone microarchitecture via micro computed tomography, and detected Gal-3, Runt-related transcription factor 2 (Runx2), and interleukin-6 (IL-6) expression in alveolar bone. Applying dual luciferase reporter assay, we explored the target binding of miR-124-3p and Gal-3. We examined osteocyte-derived exosomes with transmission electron microscopy and detected miR-124-3p, Gal-3, and IL-6 expression in exosomes. Saliva Gal-3 was increased in DM compared with controls but decreased in patients with moderate periodontitis and DM compared with those who had moderate periodontitis only. Alveolar bone mass was increased in DM and exacerbated in DM with periodontitis. Gal-3 and Runx2 were both increased in periodontitis and DM compared with controls, but decreased in DM with periodontitis compared with DM alone. MiR-124-3p targeted and inhibited Gal-3 expression in vitro. Osteocytes secreted exosomes carrying miR-124-3p, Gal-3, and IL-6, which were influenced by high glucose. These findings indicate that osteocyte-derived exosomes carrying miR-124-3p may regulate Gal-3 expression of osteoblasts, especially under high-glucose conditions, suggesting a possible mechanism for DM-related alveolar bone pathologies.

Endoplasmic reticulum stress remodels alveolar bone formation after tooth extraction.

Bone healing in tooth extraction sockets occurs in a complex environment containing saliva and many microorganisms and is affected by many factors. Endoplasmic reticulum (ER) stress affects bone metabolism, but the role of ER stress in bone healing after tooth extraction remains unclear. We utilized a rat tooth extraction model, in which we promoted wound healing by using salubrinal to regulate the ER stress response. Western blot analysis showed increased expression of p-eIF2α/eIF2α, Runx2 and alkaline phosphatase (ALP) in bone tissue, and histological assays showed irregularly arranged and new bone with more collagen fibres 14 days after tooth extraction and after modulating the degree of ER stress. Micro-CT showed that modulating ER stress to an appropriate degree increases bone filling in regards to the density in the bottom and the surrounding bone wall of the tooth extraction wounds. Transmission electron microscopy showed rough ER expansion and newly formed collagen fibrils in osteoblasts after modulating ER stress to an appropriate degree. We also used different concentrations of salubrinal to evaluate the resistance to tunicamycin-induced ER stress in an osteogenic induction environment. Salubrinal restored the tunicamycin-induced decrease in the viability of primary calvarial osteoblasts and increased the expression of Runx2 and ALP, and decreased p-eIF2α/eIF2α in a dose-dependent manner. Taken together, the results demonstrate that ER stress occurred after tooth extraction, and regulating the degree of ER stress can promote bone healing in tooth extraction sockets, providing clinical evidence for bone healing.

Associations between the codon 72 polymorphism of the TP53 gene and the risk of recurrent implantation failure.

AIM: Recurrent implantation failure (RIF) is the most common cause of unsuccessful pregnancy after assisted reproductive techniques. The tumor protein P53 (TP53) codon 72 polymorphism (G-C transversion) has been explored in susceptibility to RIF, but inconclusive results have been reported. The aim of this article is to estimate the associations between the TP53 codon 72 polymorphism and the risk of RIF. MATERIALS AND METHODS: A comprehensive search for relevant articles was conducted. The odds ratios (ORs) and 95% confidence intervals (CIs) for CC + GC versus GG, CC versus GC + GG, CC versus GG, GC versus GG genotypes, and C versus G allele, were estimated. Publication bias was explored. Statistical analyses were performed using RevMan 5.2 and Stata 11.0 software. RESULTS: A total of five case-control studies in five articles with 417 RIF cases and 325 controls were included. An overall random effect OR of 1.20 (95% CI, 0.66-2.19; P = 0.55) in the dominant model (CC + GC vs GG) was found. The results suggested that a lack of increased or decreased risks were found in individuals who carried the CC homozygote and heterozygote GC, in comparison with the homozygote GG. However, in subgroup analysis by ethnicity, a significantly increased risk was observed among Latin Americans in the dominant model (OR, 1.56; 95% CI, 1.04-2.33; P = 0.03). CONCLUSIONS: This meta-analysis shows that the TP53 codon 72 polymorphism is not associated with RIF risk in the overall population; however, associations were found in the Latin American population.

The codon 72 polymorphism of the TP53 gene and endometriosis risk: a meta-analysis.

Endometriosis is a chronic, inflammatory and common gynaecological disease. This study investigated the association between TP53 codon 72 polymorphism and the risk of endometriosis. A search for relevant articles was conducted in PubMed, Embase, CNKI, Wanfang, Weipu databases and Google Scholar. The strength of the relationships between TP53 codon 72 polymorphism and the risk of endometriosis was assessed by odds ratios (OR) and with 95% confidence intervals (CI). Sixteen case-control studies in 15 articles were included. Significant association was found in the dominant model (CC + GC versus GG) with an OR of 1.38 and 95% CI (1.14, 1.67). The results suggested that individuals who carried CC homozygote and heterozygote GC might have a 38% increased endometriosis risk when compared with the homozygote GG. In the subgroup analysis by ethnicity, significantly increased risk was observed among Asians (OR = 1.62, 95% CI = 1.18-2.23, P = 0.003) and Latin Americans (OR = 1.54, 95% CI = 1.16-2.03, P = 0.002) but not in Caucasians (OR = 1.02, 95% CI = 0.80-1.30) for the dominant model. The current meta-analysis suggested that TP53 codon 72 polymorphism was associated with the endometriosis risk, especially in Asians and Latin Americans.

The Gothenburg Trismus Questionnaire in China: Cross-cultural adaptation and measurement invariance.

OBJECTIVES: The Gothenburg Trismus Questionnaire (GTQ) is a comprehensive scale for screening and assessing trismus in head and neck (H&N) cancer and temporomandibular joint disorders (TMD) patients. This study aimed to translate and cross-culturally adapt the GTQ in China, and to test its measurement invariance. METHODS: This study comprised 278 H&N cancer, 245 TMD, and 507 control patients. Internal consistency and test-retest reliability were tested to assess the GTQ's reliability. The validity was evaluated through composite reliability (CR), average variance extracted (AVE), and correlation tests. Multi-group confirmatory factor analysis (CFA) was used to investigate the GTQ's measurement invariance across clinical status and gender. T tests were employed to compare score differences across clinical status and gender. RESULTS: The Chinese version of GTQ scale shows excellent internal consistency and test-retest reliability. The CR, AVE, and correlation values demonstrate the good validity of GTQ. The multi-group CFA supported configural invariance across clinical status but not metric invariance, while it supported strict invariance across gender. Additionally, t tests revealed that patients with H&N cancer and TMD scored higher than the control group, while males scored higher than females. CONCLUSIONS: The Chinese version of GTQ serves as an effective tool for screening and assessing trismus.

Plasticity of adipose tissues in response to fasting and refeeding declines with aging in mice.

To explore the plasticity of adipose tissues, C57BL/6J mice at the age of 1 month, 3 months, and 15 months corresponding to adolescence, adulthood, and middle-aged transitional period, respectively, were fasted and refed subsequently at different times. Body adipose tissues ratio (BATR) was calculated, the morphology of adipose tissue and the area of adipocytes were observed by histological analysis, and the mitochondria in adipocytes were observed under the transmission electron microscope. Furthermore, the expression levels of Ucp-1, Cidea, Cox7a1, Cpt-1m, Atgl, and Hsl were detected by qRT-PCR. Our results showed a significant increase in the adipocytes area and body visceral adipose tissue (VAT) ratio in all groups of mice with aging. Moreover, body mesenteric white adipose tissue (mWAT) ratio decreased the most after 72 h fasting. In the middle-aged transitional mice, the white adipocytes did not decrease until 72 h fasting, and most of them still appeared as unaffected unilocular cells. Besides, the number of mitochondria and the expression of Ucp-1, Cidea, Cox7a1, Cpt-1m, Atgl and Hsl were lower in these mice. After 72h refeeding, the body subcutaneous white adipose tissue (sWAT) ratio returned to normal, while the VAT kept decreasing. The above results indicated an impairment in adipose tissue plasticity in mice with aging, suggesting that age modulated the metabolic adaptiveness of adipose tissues in mice.

Hereditary severe insulin resistance syndrome: Pathogenesis, pathophysiology, and clinical management.

Severe insulin resistance has been linked to some of the most globally prevalent disorders, such as diabetes mellitus, nonalcoholic fatty liver disease, polycystic ovarian syndrome, and hypertension. Hereditary severe insulin resistance syndrome (H-SIRS) is a rare disorder classified into four principal categories: primary insulin receptor defects, lipodystrophies, complex syndromes, and obesity-related H-SIRS. Genes such as INSR, AKT2, TBC1D4, AGPAT2, BSCL2, CAV1, PTRF, LMNA, PPARG, PLIN1, CIDEC, LIPE, PCYT1A, MC4R, LEP, POMC, SH2B1, RECQL2, RECQL3, ALMS1, PCNT, ZMPSTE24, PIK3R1, and POLD1 have been linked to H-SIRS. Its clinical features include insulin resistance, hyperglycemia, hyperandrogenism, severe dyslipidemia, fatty liver, abnormal topography of adipose tissue, and low serum leptin and adiponectin levels. Diagnosis of H-SIRS is based on the presence of typical clinical features associated with the various H-SIRS forms and the identification of mutations in H-SIRS-linked genes by genetic testing. Diet therapy, insulin sensitization, exogenous insulin therapy, and leptin replacement therapy have widely been adopted to manage H-SIRS. The rarity of H-SIRS, its highly variable clinical presentation, refusal to be tested for genetic mutations by patients' family members who are not severely sick, unavailability of genetic testing, and testing expenses contribute to the delayed or underdiagnoses of H-SIRS. Early diagnosis facilitates early management of the condition, which results in improved glycemic control and delayed onset of diabetes and other complications related to severe insulin resistance. The use of updated genetic sequencing technologies is recommended, and long-term studies are required for genotype-phenotype differentiation and formulation of diagnostic and treatment protocols.

Kangfuxin Accelerates Extraction Socket Healing by Promoting Angiogenesis Via Upregulation of CCL2 in Stem Cells.

Kangfuxin (KFX) shows potential in wound healing, but its role in socket healing is unclear. This research finds increased bone mass, mineralization, and collagen deposition in KFX-treated mice. Mouse bone marrow mesenchymal stem cells, human periodontal ligament stem cells (hPDLSCs), and human dental pulp stem cells (hDPSCs) are treated with KFX under osteogenic induction. RNA-sequencing reveals upregulated chemokine-related genes, with a threefold increase in chemokine (C-C motif) ligand 2 (Ccl2). The conditioned medium (CM) of hPDLSCs and hDPSCs treated with KFX promotes endothelial cell migration and angiogenesis. Ccl2 knockdown abolishes CM-induced endothelial cell migration and angiogenesis, which can be reversed by recombinant CCL2 treatment. KFX-treated mice showed increased vasculature. In conclusion, KFX increases the expression of CCL2 in stem cells, promoting bone formation and mineralization in the extraction socket by inducing endothelial cell angiogenesis. © 2023 American Society for Bone and Mineral Research (ASBMR).

Potential interaction between the oral microbiota and COVID-19: a meta-analysis and bioinformatics prediction.

Objectives: The purpose of this study was to evaluate available evidence on the association between the human oral microbiota and coronavirus disease 2019 (COVID-19) and summarize relevant data obtained during the pandemic. Methods: We searched EMBASE, PubMed, and the Cochrane Library for human studies published up to October 2022. The main outcomes of the study were the differences in the diversity (α and ß) and composition of the oral microbiota at the phylum and genus levels between patients with laboratory-confirmed SARS-CoV-2 infection (CPs) and healthy controls (HCs). We used the Human Protein Atlas (HPA), Gene Expression Profiling Interactive Analysis (GEPIA) database, Protein-protein interaction (PPI) network (STRING) and Gene enrichment analysis (Metascape) to evaluate the expression of dipeptidyl peptidase 4 (DPP4) (which is the cell receptor of SARS CoV-2) in oral tissues and evaluate its correlation with viral genes or changes in the oral microbiota. Results: Out of 706 studies, a meta-analysis of 9 studies revealed a significantly lower alpha diversity (Shannon index) in CPs than in HCs (standardized mean difference (SMD): -0.53, 95% confidence intervals (95% CI): -0.97 to -0.09). Subgroup meta-analysis revealed a significantly lower alpha diversity (Shannon index) in older than younger individuals (SMD: -0.54, 95% CI: -0.86 to -0.23/SMD: -0.52, 95% CI: -1.18 to 0.14). At the genus level, the most significant changes were in Streptococcus and Neisseria, which had abundances that were significantly higher and lower in CPs than in HCs based on data obtained from six out of eleven and five out of eleven studies, respectively. DPP4 mRNA expression in the oral salivary gland was significantly lower in elderly individuals than in young individuals. Spearman correlation analysis showed that DPP4 expression was negatively correlated with the expression of viral genes. Gene enrichment analysis showed that DPP4-associated proteins were mainly enriched in biological processes, such as regulation of receptor-mediated endocytosis of viruses by host cells and bacterial invasion of epithelial cells. Conclusion: The oral microbial composition in COVID-19 patients was significantly different from that in healthy individuals, especially among elderly individuals. DPP4 may be related to viral infection and dysbiosis of the oral microbiome in elderly individuals.

Oral cavity-derived stem cells and preclinical models of jaw-bone defects for bone tissue engineering.

BACKGROUND: Jaw-bone defects caused by various diseases lead to aesthetic and functional complications, which can seriously affect the life quality of patients. Current treatments cannot fully meet the needs of reconstruction of jaw-bone defects. Thus, the research and application of bone tissue engineering are a "hot topic." As seed cells for engineering of jaw-bone tissue, oral cavity-derived stem cells have been explored and used widely. Models of jaw-bone defect are excellent tools for the study of bone defect repair in vivo. Different types of bone defect repair require different stem cells and bone defect models. This review aimed to better understand the research status of oral and maxillofacial bone regeneration. MAIN TEXT: Data were gathered from PubMed searches and references from relevant studies using the search phrases "bone" AND ("PDLSC" OR "DPSC" OR "SCAP" OR "GMSC" OR "SHED" OR "DFSC" OR "ABMSC" OR "TGPC"); ("jaw" OR "alveolar") AND "bone defect." We screened studies that focus on "bone formation of oral cavity-derived stem cells" and "jaw bone defect models," and reviewed the advantages and disadvantages of oral cavity-derived stem cells and preclinical model of jaw-bone defect models. CONCLUSION: The type of cell and animal model should be selected according to the specific research purpose and disease type. This review can provide a foundation for the selection of oral cavity-derived stem cells and defect models in tissue engineering of the jaw bone.

Long noncoding RNA BBOX1-AS1 increased radiotherapy sensitivity in colorectal cancer by stabilizing and activating PFK1.

PURPOSE: Our study explored the effect of long noncoding RNA BBOX1-AS1 on colorectal cancer (CRC) radiosensitivity in vivo and in vitro. METHODS: Differentially expressed lncRNAs in CRC were screened using a bioinformatics database and an online prediction website. The expression of BBOX1-AS1 in tissue samples was analyzed via real-time quantitative PCR (RT-qPCR). Subcellular localization of BBOX1-AS1 in CRC cells was analyzed using fluorescence in situ hybridization (FISH). The correlation between BBOX1-AS1 and PFK1 expression levels in CRC tissues was analyzed via Pearson's correlation coefficient. The effect of BBOX1-AS1 on PFK1 stability was investigated using RNA and protein stability testing. RNA Binding Protein Immunoprecipitation (RIP) and RNA pull-down assays were used to confirm the binding of BBOX1-AS1 to PFK1. RESULTS: BBOX1-AS1 was highly expressed in CRC and associated with poor prognosis. Similarly, it was highly expressed in CRC tissues and CRC cell lines. In addition, BBOX1-AS1 promoted the proliferation, invasion, migration, and glycolysis of CRC cells and inhibited apoptosis. RIP and RNA pull-down experiments confirmed that BBOX1-AS1 bound to PFK1. RNA stability and protein stability experiments showed that BBOX1-AS1 affected the stability of PFK1 mRNA and protein. Furthermore, we confirmed that BBOX1-AS1 increased radiation resistance through the regulation of PFK1 expression. CONCLUSIONS: BBOX1-AS1 promoted the proliferation, invasion, migration, and glycolysis of CRC cells through stabilization of the expression of PFK1. BBOX1-AS1 also inhibited CRC cell apoptosis and increased radiotherapy resistance in CRC cells.

Prognostic biomarker GSTK1 in head and neck squamous cell carcinoma and its correlation with immune infiltration and DNA methylation.

Background: Glutathione S-transferase kappa 1 (GSTK1) is critical in sarcoma and breast cancer (BRCA) development. However, the clinical significance of GSTK1 in head and neck squamous cell carcinoma (HNSC) remains unclear. This study is the first investigation into the role of GSTK1 in HNSC. Methods: All original data were downloaded from the Cancer Genome Atlas (TCGA) dataset and verified by R Base Package 4.2.0. The expression of GSTK1 in various cancers was explored with TIMER and TCGA databases. Prognostic value of GSTK1 was analyzed via survival module of Kaplan-Meier plotter and Human Protein Atlas database and Cox regression analysis. The association between GSTK1 and clinical features was evaluated by Wilcoxon signed-rank test and logistic regression analysis. The relationship between GSTK1 and immune infiltration and methylation level was further explored. The expression of GSTK1 and its correlation with immune cell infiltration was verified by Immunohistochemical staining (IHC). Results: GSTK1 was lower in HNSC, BRCA, Lung squamous cell carcinoma, and Thyroid carcinoma than in para-carcinoma. Low GSTK1 expression was associated with worse overall survival in Bladder urothelial carcinoma, Kidney renal papillary cell carcinoma, BRCA, and HNSC. However, only in BRCA and HNSC, GSTK1 expression in tumors was lower than that in normal tissues. Cox regression analyses confirmed that GSKT1 was an independent prognostic factor of overall survival in HNSC patients. The decrease in GSTK1 expression in HNSC was significantly correlated with high T stage and smoker history. IHC showed that the expression level of GSTK1 in HNSC was lower than that in para-carcinoma. In addition, GSEA showed that three pathways related to immune infiltration were positively correlated, while two pathways related to DNA methylation were negatively correlated with expression of GSTK1. Further analysis showed that GSTK1 was moderately positively correlated with the infiltration level of T cells and Cytotoxic cells, which was further confirmed by IHC. The methylation level of GSTK1 was associated with prognosis in patients with HNSC. Conclusion: Low GSTK1 expression may be a potential molecular marker for poor prognosis in HNSC and provide new insight for the development of diagnostic marker or therapeutic target.

Association of Sex Hormones and Fat Distribution in Men with Different Obese and Metabolic Statuses.

PURPOSE: Although several studies have explored the association of sex hormones with glucose metabolism, the association between sex hormones and body fat distribution, which is closely related to insulin resistance, has not been fully elucidated. We have tried to explore the relationship of testosterone (T) and estradiol (E2) with visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) mass in Chinese men with different obese and metabolic statuses. PATIENTS AND METHODS: A total of 128 men from the Health Management Center of the Second Xiangya Hospital, Central South University were collected and grouped in accordance with their obese and metabolic syndrome (MS) statuses: metabolically healthy non-overweight/obese men (MHNO), metabolically healthy overweight/obese men (MHO) and metabolically unhealthy overweight/obese men (MUO). Multiple regression analyses were performed to estimate contributions of sex hormones levels to the variations of body fat distribution and the contributions of body fat distribution to the variations of sex hormone levels. RESULTS: With fat mass parameters as independent variables, SAT had a strong negative association with T in MHNO (ß = -2.772, P = 0.034), VAT was positively correlated with E2 in MHO (ß = 22.269, P = 0.009), and SAT was negatively associated with T in MUO (ß = -3.315, P = 0.010). With sex hormones as independent variables, E2 positively correlated with VAT (ß = -176.259, P = 0.048), while T negatively correlated with VAT in MHO (ß = 183.150, P = 0.029). In MUO, an inverse association of T with SAT was observed (ß = -213.689, P = 0.021). CONCLUSION: E2 and VAT had a mutual influence, thus resulting in a vicious circle, and the negative correlation between T and VAT may be related to the decrease of the MS occurrence in the MHO group. There were bi-directional relationships between sex hormones and fat distribution in men with different obese and metabolic statuses. TRIAL REGISTRATION: Chinese Clinical Trial Registry, ChiCTR-EOC-16010194. Retrospectively registered.

A novel antimicrobial peptide derived from human BPIFA1 protein protects against Candida albicans infection.

Bactericidal/permeability-increasing fold containing family A, member 1 (BPIFA1) is an innate immunity defense protein. Our previous studies proved its antibacterial and antiviral effects, but its role in fungi remains unknown. The study aimed to identify antifungal peptides (AFP) derived from BPIFA1, and three antimicrobial peptides (AMP1-3) were designed. The antifungal effects were proved by growth inhibition assay. AMP3 activity was confirmed by germ tube growth experiment and XTT assay. Its effects on cell wall and membrane of Candida albicans were assessed by tannic acid and Annexin V-FITC/PI double staining, respectively. Additionally, scanning electron microscope (SEM) and transmission electron microscopy (TEM) were used for morphological and ultrastructural observation. The expression of ALS1, EAP1, and SUN41 was tested by qPCR. Ultimately, three AMPs could fight against C. albicans in vitro, and AMP3 was highly effective. It functioned by destroying the integrity of cell wall and normal structure of cell membrane. It also inhibited biofilm formation of C. albicans. In addition, AMP3 down-regulated the expression of ALS1, EAP1, and SUN41, those are known to be involved in virulence of C. albicans. Altogether, the study reported successful development of a novel AFP, which could be used as a new strategy for antifungal therapy.

Spheroid co-culture of BMSCs with osteocytes yields ring-shaped bone-like tissue that enhances alveolar bone regeneration.

Oral and maxillofacial bone defects severely impair appearance and function, and bioactive materials are urgently needed for bone regeneration. Here, we spheroid co-cultured green fluorescent protein (GFP)-labeled bone marrow stromal cells (BMSCs) and osteocyte-like MLO-Y4 cells in different ratios (3:1, 2:1, 1:1, 1:2, 1:3) or as monoculture. Bone-like tissue was formed in the 3:1, 2:1, and 1:1 co-cultures and MLO-Y4 monoculture. We found a continuous dense calcium phosphate structure and spherical calcium phosphate similar to mouse femur with the 3:1, 2:1, and 1:1 co-cultures, along with GFP-positive osteocyte-like cells encircled by an osteoid-like matrix similar to cortical bone. Flake-like calcium phosphate, which is more mature than spherical calcium phosphate, was found with the 3:1 and 2:1 co-cultures. Phosphorus and calcium signals were highest with 3:1 co-culture, and this bone-like tissue was ring-shaped. In a murine tooth extraction model, implantation of the ring-shaped bone-like tissue yielded more bone mass, osteoid and mineralized bone, and collagen versus no implantation. This tissue fabricated by spheroid co-culturing BMSCs with osteocytes yields an internal structure and mineral composition similar to mouse femur and could promote bone formation and maturation, accelerating regeneration. These findings open the way to new strategies in bone tissue engineering.