Identification and characterization of the ferroptosis-related ceRNA network in irreversible pulpitis.

Background: The role of ferroptosis in irreversible pulpitis (IP) remains unclear. The competing endogenous RNA (ceRNA) theory that has been widely investigated is rarely used studied in IP. Hub lncRNAs selected from a ceRNA network may provide a novel hypothesis for the interaction of ferroptosis and IP. Methods: Differentially expressed genes (DEGs) were intersected with 484 ferroptosis markers to identify differentially expressed ferroptosis-related genes (DE-FRGs). Functional analysis and protein-protein interaction (PPI) networks were constructed to reveal the functions of DE-FRGs. Then, coexpression analyses were conducted between DE-FRGs and DElncRNAs to define ferroptosis-related DElncRNAs (FR-DElncRNAs). Predictions of DE-FRG- and FR-DElncRNA-related miRNAs were obtained, and members of both groups were selected. Additionally, two ceRNA networks consisting of FR-DElncRNAs, miRNAs and DE-FRGs from upregulated and downregulated groups were built. Finally, the hub lncRNAs of the ceRNA networks were used for immuno-infiltration analysis and qPCR verification. Results: According to the results of PCA and clustering analysis, 5 inflamed and 5 healthy pulp tissue samples were selected for analysis. The intersection of DEGs with 484 ferroptosis marker genes identified 72 DE-FRGs. The response to stimulus, cellular process, signaling, localization, and biological regulation pathways related to DE-FRGs were enriched. In total, 161 downregulated and 40 upregulated FR-DElncRNAs were chosen by coexpression analysis for further investigation. The MultimiR package and starBase were used to predict miRNAs of DE-FRGs and FR-DElncRNAs, respectively. The upregulated ceRNA network contained 2 FR-DElncRNAs (↑), 19 miRNAs (↓) and 22 DE-FRGs (↑). The downregulated network contained 44 FR-DElncRNAs (↓), 251 miRNAs (↑) and 10 DE-FRGs (↓). Six hub lncRNAs were identified based on the MCC method (LUCAT1 and AC106897.1 ↑; LINC00943, AL583810.1, AC068888.1, and AC125257.1↓). In addition, strong relationships between hub lncRNAs and immune cells were shown by immune infiltration analysis. Finally, validated by qPCR assays of the pulp tissue of IP patients, the expression levels in clinical samples were consistent with the microarray data. Conclusion: Two ceRNA networks were comprehensively constructed, and 6 hub lncRNAs were identified. These genes provide novel insights into the relationship between ferroptosis and IP. Intriguingly, the LINC00943/hsa-miR-29a-3p/PDK4 axis was deemed to be the key node in this network.

Establishment and validation of the autophagy-related ceRNA network in irreversible pulpitis.

BACKGROUND: The molecular mechanisms underlying the onset and progression of irreversible pulpitis have been studied for decades. Many studies have indicated a potential correlation between autophagy and this disease. Against the background of the competing endogenous RNA (ceRNA) theory, protein-coding RNA functions are linked with long noncoding RNAs (lncRNAs) and microRNAs (miRNAs). This mechanism has been widely studied in various fields but has rarely been reported in the context of irreversible pulpitis. The hub genes selected under this theory may represent the key to the interaction between autophagy and irreversible pulpitis. RESULTS: Filtering and differential expression analyses of the GSE92681 dataset, which contains data from 7 inflamed and 5 healthy pulp tissue samples, were conducted. The results were intersected with autophagy-related genes (ARGs), and 36 differentially expressed ARGs (DE-ARGs) were identified. Functional enrichment analysis and construction of the protein‒protein interaction (PPI) network of DE-ARGs were performed. Coexpression analysis was conducted between differentially expressed lncRNAs (DElncRNAs) and DE-ARGs, and 151 downregulated and 59 upregulated autophagy-related DElncRNAs (AR-DElncRNAs) were identified. StarBase and multiMiR were then used to predict related microRNAs of AR-DElncRNAs and DE-ARGs, respectively. We established ceRNA networks including 9 hub lncRNAs (HCP5 and AC112496.1 ↑; FENDRR, AC099850.1, ZSWIM8-AS1, DLX6-AS1, LAMTOR5-AS1, TMEM161B-AS1 and AC145207.5 ↓), which were validated by a qRT‒PCR analysis of pulp tissue from patients with irreversible pulpitis. CONCLUSION: We constructed two networks consisting of 9 hub lncRNAs based on the comprehensive identification of autophagy-related ceRNAs. This study may provide novel insights into the interactive relationship between autophagy and irreversible pulpitis and identifies several lncRNAs that may serve as potential biological markers.

Role of microglia autophagy and mitophagy in age-related neurodegenerative diseases.

Microglia, characterized by responding to damage, regulating the secretion of soluble inflammatory mediators, and engulfing specific segments in the central nervous system (CNS), function as key immune cells in the CNS. Emerging evidence suggests that microglia coordinate the inflammatory responses in CNS system and play a pivotal role in the pathogenesis of age-related neurodegenerative diseases (NDDs). Remarkably, microglia autophagy participates in the regulation of subcellular substances, which includes the degradation of misfolded proteins and other harmful constituents produced by neurons. Therefore, microglia autophagy regulates neuronal homeostasis maintenance and process of neuroinflammation. In this review, we aimed at highlighting the pivotal role of microglia autophagy in the pathogenesis of age-related NDDs. Besides the mechanistic process and the co-interaction between microglia autophagy and different kinds of NDDs, we also emphasized potential therapeutic agents and approaches that could be utilized at the onset and progression of these diseases through modulating microglia autophagy, including promising nanomedicines. Our review provides a valuable reference for subsequent studies focusing on treatments of neurodegenerative disorders. The exploration of microglia autophagy and the development of nanomedicines greatly enhances current understanding of NDDs.

Construction of ceRNA and m6A-related lncRNA networks associated with anti-inflammation of AdipoAI.

Background: Adiponectin (APN) is an endogenous adipokine secreted from adipocytes that exerts anti-inflammatory properties. AdipoAI is an orally active adiponectin receptor agonist identified by our group that can emulate APN's anti-inflammatory properties through mechanisms that are not fully understood. LncRNAs, a type of noncoding RNA more than 200 bp in length, have been demonstrated to have abundant biological functions, including in anti-inflammatory responses. Materials and Result: In the current study, we performed a lncRNA microarray in LPS-induced Raw264.7 cells that were prestimulated with AdipoAI and screened 110 DElncRNAs and 190 DEmRNAs. Enrichment analyses were conducted on total mRNAs and DEmRNAs, including GSVA, ssGSEA, GO/KEGG, GSEA, and PPI analysis. Among all these processes, endocytosis was significantly enriched. A coexpression analysis was built based on DElncRNAs and DEmRNAs. Then, using TargetScan and miRwalk to predict related microRNAs of DElncRNAs and DEmRNAs, respectively, we established competing endogenous RNA (ceRNA) networks including 54 mRNAs from 8 GO items. Furthermore, 33 m6A methylation-related marker genes were obtained from a previous study and used for the construction of an m6A-related lncRNA network by coexpression analysis. We identified FTO as the hub gene of the network and 14 lncRNAs that interacted with it. The expression levels of 10 lncRNAs selected from ceRNA and FTO-related lncRNA networks were validated with qRT‒PCR. Finally, macrophage phenotype scores showed that AdipoAI could attenuate the M2b and M2c polarization of macrophages and correlate with the above lncRNAs. Conclusion: Our work reveals that lncRNAs might be involved in the anti-inflammation process of AdipoAI in LPS-induced macrophages through the ceRNA network and the epigenetic regulation of m6A. Mechanistically, these lncRNAs associated with AdipoAI might be related to endocytosis and polarization in macrophages and provide new candidates for the anti-inflammatory application of APN and its receptor agonist.

[Size distribution characteristics of particulate matter in the top areas of coke oven].

OBJECTIVE: To systematically evaluate the environmental exposure information of coke oven workers, we investigated the concentration and size distribution characteristics of the particle matter (PM) in the top working area of coke oven. METHODS: The aerodynamic particle sizer spectrometer was employed to collect the concentration and size distribution information of PM at a top working area. The PM was divided into PM ≤ 1.0 µm, 1.0 µm < PM ≤ 2.5 µm, 2.5 µm < PM ≤ 5.0 µm, 5.0 µm < PM ≤ 10.0 µm and PM>10.0 µm based on their aerodynamic diameters. The number concentration, surface area concentration, and mass concentration were analyzed between different groups. We also conducted the correlation analysis on these parameters among groups. RESULTS: We found the number and surface area concentration of top area particulate was negatively correlated with particle size, but mass concentration curve showed bimodal type with higher point at PM = 1.0 µm and PM = 5.0 µm. The average number concentration of total particulate matter in the top working area was 661.27 number/cm³, surface area concentration was 523.92 µm²/cm³, and mass concentration was 0.12 mg/m³. The most number of particulate matter is not more than 1 µm (PM(1.0)), and its number concentration and surface area concentration accounted for 96.85% and 67.01% of the total particles respectively. In the correlation analysis, different particle size correlated with the total particulate matter differently. And the characteristic parameters of PM2.5 cannot fully reflect the total information of particles. CONCLUSION: The main particulate matter pollutants in the top working area of coke oven is PM1.0, and it with PM(5.0) can account for a large proportion in the mass concentration of PM. It suggest that PM1.0 and PM(5.0) should be considered for occupational health surveillance on the particulate matter in the top area of coke oven.

[Study on the pulmonary inflammatory effects induced by inhalation exposure to nanoscale carbon black aerosol in mice].

OBJECTIVE: To explore the carbon black induced effects of lung morphology and pro-inflammation in mice, based on the carbon black aerosol dynamic inhalation exposure model. METHODS: The carbon black aerosol generated by dynamic inhalation device was imported exposure chamber to mice. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to observe the characters of carbon black. Sixty 9-week-old male BALB/c mice were randomly divided into two control groups, 7 d exposure group and 14 d exposure group. The numbers of four groups of animals were 15, respectively. Mice were exposed to carbon black in the inhalation chamber at (29.33 ± 9.10) mg/m(3) for 6 h/d for continuous exposure 7 d and 14 d, respectively. After 7 d and 14 d exposure, the mice were sacrificed after the last exposure for 24 h. Control mice were killed at 7 d and 14 d. The trachea, lungs, liver, kidneys, and spleen tissues were separated and weighted. Hematoxylin and eosin (HE) staining was used to observe pathological changes of lung by light microscopy. Pulmonary interleukin-8 (IL-8) expression was analyzed by immunohistochemistry. Transmission electron microscopy was used to observe the ultra structure of lung tissue. RESULTS: After 14 d exposure carbon black, the lung coefficient was increased in exposure group compared with control (0.61 ± 0.03 vs 0.79 ± 0.06, t = 6.26, P < 0.01). The spleen coefficient were higher than control(0.39 ± 0.04 vs 0.51 ± 0.06, t = 4.23, P < 0.01) . Other organ coefficients were no significant difference between CB group and control group.Histopathology displayed carbon black particles were deposited in the alveoli and lung bronchial wall in 7 d and 14 d groups. The black carbon particles were deposited within the lung tissue of mice in 14 d group. There were cilia damage, serious damage to the alveolar wall, inflammatory cell infiltration and more hyperemia in 14 d group. Immunohistochemistry showed the level of IL-8 in 7 d (0.272 ± 0.011) and 14 d (0.422 ± 0.065) exposure group were higher than control group in 14 d (0.188 ± 0.041) , F = 31.89, P < 0.01. TEM showed that the lung tissue vision was clear and organelle integrity in the control group. The particles appeared in lung tissue macrophage lysosomes in exposure group, the electron density was consistent with the carbon black particles. CONCLUSION: The dynamic carbon black particles exposure can affect the lung and spleen coefficient, damage integrity of lung morphology and induce inflammation in mice.

[Relationship among pathological changes in Liver tissues and level of serum HBV DNA, HBeAg and ALT of 194 patients with chronic hepatitis B].

OBJECTIVE: To study the relationships among pathological and immunohistochemical changes in liver tissues, and the HBeAg, HBV DNA, ALT level in the patients with chronic hepatitis B. METHODS: Pathological and immunohistochemical examinations of liver tissue liver function tests, serum HBV and HBV DNA detection were performed in 194 patients with chronic hepatitis B. RESULTS: There was significant difference between the serum HBeAg positive group and the negative group in G2, G3-4 S2, S3-4 in liver tissues; The serum HBV DNA level of the groups S0 and S1-4, and the hepatic activity index between the groups G0-1 and G2-4 were significantly different. And the hepatic HBcAg positive group and HBcAg negative group were significantly different too. There was no significant difference between the HBsAg level in liver tissues as "+" group and the "++ - +++" group. The pathological diagnosis as S1 or (and) G2 is respectively 28.57%, 53.33%, 80.15%, 77.88% among the four groups with normal-mild-moderate-severe elevated serum ALT level. CONCLUSION: Serum HBV DNA correlated with HBcAg expression in liver tissue; the HBsAg level in liver tissues have no relationship with the serum HBV DNA level. The patients with low serum HBV DNA level may have high index of hepatic activity and hepatic fibrosis. Asymptomatic carriers and patients with low serum ALT level should be encouraged to accept liver biopsy. It can determine the degree of liver inflammation and fibrosis and timing of treatment.