Molecular and cellular consequences of mitochondrial DNA double-stranded breaks.

Mitochondria are subcellular organelles essential for life. Beyond their role in producing energy, mitochondria govern various physiological mechanisms, encompassing energy generation, metabolic processes, apoptotic events, and immune responses. Mitochondria also contain genetic material that is susceptible to various forms of damage. Mitochondrial double-stranded breaks (DSB) are toxic lesions that the nucleus repairs promptly. Nevertheless, the significance of DSB repair in mammalian mitochondria is controversial. This review presents an updated view of the available research on the consequences of mitochondrial DNA DSB from the molecular to the cellular level. We discuss the crucial function of mitochondrial DNA damage in regulating processes such as senescence, integrated stress response, and innate immunity. Lastly, we discuss the potential role of mitochondrial DNA DSB in mediating the cellular consequences of ionizing radiations, the standard of care in treating solid tumors.

The fate of antibiotic resistance genes in wastewater containing microalgae treated by chlorination, ultra-violet, and Fenton reaction.

Antibiotic resistance genes (ARGs) and bacteria (ARBs) in the effluent of wastewater treatment plants (WWTPs) are of utmost importance for the dissemination of ARGs in natural aquatic environments. Therefore, there is an urgent need for effective technologies to eliminate WWTP ARGs/ARBs and mitigate the associated risks posed by the discharged ARG in aquatic environments. To test the effective technology for eliminating ARGs/ARBs, we compared the removal of ARGs and ARBs by three different tertiary treatments, namely ultra-violet (UV) disinfection, chlorination disinfection, and Fenton oxidation. Then, the treated wastewater was co-cultured with Chlorella vulgaris (representative of aquatic biota) to investigate the fate of discharged ARGs into the aquatic environment. The results demonstrated that chlorination (at a chlorine concentration of 15 mg/L) and Fenton (at pH 2.73, with 0.005 mol/L Fe2+ and 0.0025 mol/L H2O2) treatment showed higher efficacy in ARG removal (1.8 - 4.17 logs) than UV treatment (15 min) (1.29 - 3.87 logs). Moreover, chlorine at 15 mg/L and Fenton treatment effectively suppressed ARB regeneration while UV treatment for 15 min could not. Regardless of treatments tested in this study, the input of treated wastewater to the Chlorella system increased the number of ARGs and mobile genetic elements (MGEs), indicating the potential risk of ARG dissemination associated with WWTP discharge. Among the wastewater-Chlorella co-culture systems, chlorination resulted in less of an increase in the number of ARGs and MGEs compared to Fenton and UV treatment. When comparing the wastewater systems to the co-culture systems, it was observed that Chlorella vulgaris reduced the number of ARGs and MGEs in chlorination and UV-treated wastewater; however, Chlorella vulgaris promoted ARG survival in Fenton-treated water, suggesting that aquatic microalgae might act as a barrier to ARG dissemination. Overall, chlorination treatment not only effectively removes ARGs and inhibits ARB regeneration but also shows a lower risk of ARG dissemination. Therefore, chlorination is recommended for practical application in controlling the spread of discharged ARGs from WWTP effluent in natural aquatic environments.

Nitrogen Doped Porous Biochar/ß-CD-MOFs Heterostructures: Bi-Functional Material for Highly Sensitive Electrochemical Detection and Removal of Acetaminophen.

Acetaminophen (AC) is one of the most common over-the-counter drugs, and its pollutant in groundwater has attracted more attention due to its serious risk to human health. Currently, the research on AC is mainly focused on its detection, but few are concerned about its removal. In this work, for the first time, nitrogen-doped Soulangeana sepals derived biochar/ß-cyclodextrin-Metal-organic frameworks (N-SC/ß-CD-MOFs) composite was proposed for the simultaneous efficient removal and detection of AC. N-SC/ß-CD-MOFs combined the properties of host-guest recognition of ß-CD-MOFs and porous structure, high porosity, and large surface area of N-SC. Their synergies endowed N-SC/ß-CD-MOFs with a high adsorption capacity toward AC, which was up to 66.43 mg/g. The adsorption type of AC on the surface of N-SC/ß-CD-MOFs conformed to the Langmuir adsorption model, and the study of the adsorption mechanism showed that AC adsorption on N-SC was mainly achieved through hydrogen bonding. In addition, the high conductivity, large specific surface area and abundant active sites of N-SC/ß-CD-MOFs were of great significance to the high-performance detection of AC. Accordingly, the sensor prepared with N-SC/ß-CD-MOFs presented a wide linear range (1.0-30.0 µM) and a low limit of detection of 0.3 nM (S/N = 3). These excellent performances demonstrate that N-SC/ß-CD-MOFs could act as an efficient dual-functional material for the detection and removal of AC.

NF-E2-Related Factor 2 (Nrf2) Ameliorates Radiation-Induced Skin Injury.

Radiation-induced skin injury (RISI) commonly occur in cancer patients who received radiotherapy and is one of the first clinical symptoms after suffering from nuclear exposure. Oxidative damage is the major causes of RISI. Nuclear factor erythroid 2-related factor 2 (Nrf2) is considered as a key mediator of the cellular antioxidant response. However, whether Nrf2 can alleviate RISI after high-dose irradiation remains unknown. In this study, we demonstrated that Nrf2-deficient (Nrf2 -/-) mice were susceptible to high-dose irradiation and adenovirus-mediated overexpression of Nrf2 (ad-Nrf2) protected against radiation in skin cells. Overexpression of Nrf2 attenuated the severity of skin injury after high-dose electron beam irradiation. To uncover the mechanisms of Nrf2 involved in RISI, mRNA sequencing technology was performed to analyze the mRNA expression profiles of Ad-Nrf2 skin cells following radiation. The results revealed that a total of 127 genes were significantly changed, 55 genes were upregulated, and 72 genes were downregulated after Nrf2 overexpression. GSEA showed that Nrf2 was associated with positive regulation of genes involved in the reactive oxygen species pathway after radiation. Taken together, this study illustrated the role of Nrf2 in RISI and provided potentially strategies for ameliorating RISI.

Thoracic vertebra fixation with a novel screw-plate system based on computed tomography imaging and finite element method.

BACKGROUND AND OBJECTIVE: The traditional pedicle screw-rod internal fixation system has been widely used for thoracic diseases in clinical practice, but its high profile increases the damage to soft tissue, leading to long-term intractable back stiffness. The purpose of this study is to compare biomechanical advantages between the new spine pedicle screw-plate internal fixation system and traditional pedicle screw-rod internal fixation system using finite element analysis. METHODS: Based on computed tomography (CT), four three-dimensional finite element models of T7-T9 were constructed. The downward concentrated force of 150 N and the moment of 5 Nm was applied to the models to simulate six physiological activities, including flexion, extension, left and right lateral bending, left and right axial torsion. The maximum displacement, range of motion (ROM) and maximum stress of the two models in six physiological activities, was measured to evaluate the biomechanical advantages of the novel pedicle screw-plate internal fixation system. RESULTS: The novel pedicle screw-plate internal fixation system has a lower profile than the traditional pedicle screw-rod internal fixation system. With regards to the stability, the maximum displacement of the models of two internal fixation systems decreased by 56.2%-91.4% under the six motion status when comparing with the unstable model. Meanwhile, the ROM remained unchanged between the two models of internal fixation systems besides the left lateral bending. However, there is no significant difference in the ROM between the models of the two internal fixation systems in left lateral bending motion (P = 0.203). In terms of the strength, the maximum stress in the model with the new pedicle screw-plate internal fixation system was higher than that of model with the traditional pedicle screw-rod internal fixation system in every motion status but left and right lateral bending motion. CONCLUSIONS: The novel pedicle screw-plate internal fixation system has lower profile in orthopedics and higher strength, However, it has no disadvantage when comparing with the traditional pedicle screw-rod internal fixation system in terms of the stability. In summary, we suggest that the novel spine pedicle screw-plate system can be used as a new internal fixation and provide better comfort for patients.

Modelling tri-cortical pedicle screw fixation in thoracic vertebrae under osteoporotic condition: A finite element analysis based on computed tomography.

BACKGROUND AND OBJECTIVE: The technique of tri-cortical pedicle screw (TCPS) has been used to improve the anchoring strength in the sacral vertebrae. However, no studies have reported their application in the thoracic vertebrae. Our research is aimed to assess the stability and strength of the TCPS in thoracic vertebrae under osteoporotic condition by three dimensional (3D) finite element method on the basis of medical image reconstruction using computed tomography (CT), and verifying its effectiveness in clinical application. MATERIALS AND METHODS: The 3D finite element models were constructed using Mimcs to transfer two dimensional CT images into 3D models by marching cubes algorithm of six-partition. Six physiological activities were simulated in 3D finite element models. Compared with the strength and stability of the uni-cortical pedicle screw (UCPS) and bi-cortical pedicle screw (BCPS), the effectiveness of TCPS was assessed. The stress distribution and maximum stress were measured to evaluate the strength. The maximum displacement and the range of motion were analysed to assessed the stability. EXPERIMENTAL RESULTS: Four geometrically accurate and nonlinear T7-T9 finite element models were constructed successfully by 3D finite element method based on the CT images. Three kinds of internal fixation methods in the osteoporotic thoracic vertebral body can improved the maximum stress, decrease the maximum displacement and range of motion in six physiological activities. The range of motion and maximum displacement of TCPS decreased more significantly than that of UCPS and BCPS. The maximum von Mises stress of TCPS was minimum and UCPS was maximum under the condition of extension, right lateral bending, left rotation and right rotation. CONCLUSIONS: Effectively, TCPS can provide better stability and strength than that of UCPS and BCPS techniques in the osteoporotic thoracic vertebrae. In practice, the technique of TCPS can be applied in the osteoporotic thoracic vertebral body to enhance the griping strength of the screws and reduce the risk of pedicle screw loosening. However, further cadaver experiments and more biomechanical analysis are necessary to confirmed our findings.

Identification of microRNA-181 as a promising biomarker for predicting the poor survival in colorectal cancer.

BACKGROUND: A series of studies have investigated the vital role of microRNA-181 (miR-181) in the initiation and development of colorectal cancer (CRC), and demonstrated that it might be associated with the prognosis of CRC. However, inconsistent findings have hindered its clinical application. METHODS: A comprehensive meta-analysis and an integrative bioinformatics analysis were carried out for concluding current available evidence, clarifying the preliminary prognostic value and unfolding the underlying biological function of miR-181 in CRC patients. RESULTS: The findings revealed that elevated expression levels of miR-181 were associated with significantly poorer overall survival rates (HR = 1.75, 95% CI: 1.26-2.43, P < .05). Meanwhile, the target genes of miR-181 were identified and enriched into several important gene ontology (GO) categories and signaling pathways including miRNAs in cancer, pathways in cancer, proteoglycans in cancer, colorectal cancer, FoxO signaling pathway, PI3K-Akt signaling pathway, VEGF signaling pathway, HIF-1 signaling pathway, mTOR signaling pathway, and cAMP signaling pathway, which were confirmed highly involved in the initiation and progression of CRC. In addition, the protein-protein interaction (PPI) networks were set up by miR-181 targets to screen hub nodes and significant modules, which were also considerably associated with the molecular pathogenesis of CRC. CONCLUSIONS: The present study demonstrated that miR-181 could be a promising biomarker with predictive value for prognosis for CRC patients. However, future studies comprising large cohorts from multicenter are warranted to further investigate the biomarker value of miR-181.

GPR39 agonist TC-G 1008 promotes osteoblast differentiation and mineralization in MC3T3-E1 cells.

Osteoporosis-related bone fracture and falls have a severe impact on patients' daily lives. Osteoblasts are bone-building cells that play a vital role in bone formation and remodeling. Imbalanced osteoblast differentiation could lead to osteoporosis. GPR39 is an orphan G protein-coupled receptor that mediates metabolic pathways. In this study, we show that GPR39 is expressed in MC3T3-E1 cells. Osteoblast differentiation culture media induces GPR39, suggesting that GPR39 is a differentiation-responsive factor. Activation of GPR39 using its selective agonist TC-G 1008 induces alkaline phosphatase (ALP), osteocalcin (OCN), and type I collagen (Col-I) expression, and increases cellular ALP activity and calcium deposition, implying that GPR activation promotes cells toward osteoblast differentiation. Treatment with TC-G 1008 also increases Runx-2 expression and AMPK activation. However, the inhibition of AMPK by Compound C abolished TC-G 1008-mediated ALP, OCN, and Col-I induction, and reduces ALP activity and cellular calcium deposition as well as Runx-2 induction. These data indicate that TC-G 1008-mediated GPR39 activation involves AMPK-mediated Runx-2 induction. In summary, our study uncovers a new role of GPR39 activation in osteoblast differentiation, implying that GPR39 could be a promising therapeutic target for osteoporosis.

Metformin suppresses gastric cancer progression through calmodulin­like protein 3 secreted from tumor­associated fibroblasts.

Gastric cancer is one of the most common malignant tumor types worldwide, with a high morbidity and associated mortality. The interaction between gastric cancer cells and their microenvironment has a significant role in their maintenance and progression. Gastric tumor­associated fibroblasts (TAFs) are among the major regulators of the gastric cancer microenvironment. Metformin, a classical anti­diabetic drug, is known to prevent cancer progression. However, the effect of metformin on gastric TAFs and TAF­associated cancer progression has remained to be elucidated. In the present study, TAFs were isolated from gastric cancer patients, pre­treated with metformin and then co­cultured with gastric cancer cell lines. It was demonstrated that pre­treatment with 200 µM metformin reduced the stimulatory effect of TAFs on the proliferation of gastric cancer cells in co­culture, suggesting that metformin impairs the tumor­promoting role of TAFs. Using tandem mass tags­based quantitative proteomic analysis, it was identified that metformin significantly affected the secretion of 32 proteins (14 upregulated and 18 downregulated) in the culture medium of gastric TAFs. Among these proteins, calmodulin­like protein 3 (Calml3) was 2.88­fold upregulated in the culture medium of gastric TAFs after metformin treatment and a further experiment using recombinant Calml3 indicated its suppressive effect on the clonogenicity of gastric cancer cells. It was concluded that metformin suppresses gastric cancer through stimulating Calml3 secretion from TAFs, which represents a novel anticancer mechanism of metformin.

Downregulation of HuR Inhibits the Progression of Esophageal Cancer through Interleukin-18.

PURPOSE: The purpose of this study was to investigate the effect of human antigen R (HuR) downregulation and the potential target genes of HuR on the progression of esophageal squamous cell carcinoma (ESCC). MATERIALS AND METHODS: In this study, a proteomics assay was used to detect the expression of proteins after HuR downregulation, and a luciferase assay was used to detect the potential presence of a HuR binding site on the 3'-untranslated region (3'-UTR) of interleukin 18 (IL-18). In addition, colony formation assay, MTT, EdU incorporation assay, Western blot, flow cytometry, immunohistochemistry, transwell invasion assay, and wound healing assay were used. RESULTS: In the present study, we found that the expression of both HuR protein and mRNA levels were higher in tumor tissues than in the adjacent tissues. HuR downregulation significantly suppressed cell proliferation. In addition, the metastasis of esophageal cancer cells was inhibited, while the expression of E-cadherin was increased and the expression of matrix metalloproteinase (MMP) 2, MMP9, and vimentin was decreased after HuR knockdown. Moreover, silencing of HuR disturbed the cell cycle of ESCC cells mainly by inducing G1 arrest. Furthermore, proteomics analysis showed that downregulation of HuR in TE-1 cells resulted in 100 upregulated and 122 downregulated proteins, including IL-18 as a significantly upregulated protein. The expression of IL-18 was inversely regulated by HuR. IL-18 expression was decreased in ESCC tissues, and exogenous IL-18 significantly inhibited the proliferation and metastasis of ESCC cells. The 3'-UTR of IL-18 harbored a HuR binding site, as shown by an in vitro luciferase assay. CONCLUSION: HuR plays an important role in the progression of esophageal carcinoma by targeting IL-18, which may be a potential therapeutic target for the treatment of ESCC.

Metformin Sensitizes Non-small Cell Lung Cancer Cells to an Epigallocatechin-3-Gallate (EGCG) Treatment by Suppressing the Nrf2/HO-1 Signaling Pathway.

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. (-)-Epigallocatechin-3-gallate (EGCG), a major polyphenol in green tea, is widely studied as a cancer chemopreventive agent with potential anti-cancer effects. The NF-E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway is considered to mediate cellular resistance to EGCG. Metformin, a classical antidiabetic drug, has been shown to prevent cancer progression. Researchers have not reported whether metformin potentiates the anti-cancer efficacy of EGCG. In this study, metformin inhibited HO-1 expression and augmented the anti-tumor effect of EGCG. Metformin also enhanced ROS (reactive oxygen species) generation induced by EGCG (100 µM), subsequently resulting in apoptosis. Based on the results of the in vivo study, size of xenografts treated with the combination of metformin and EGCG was smaller than other groups. Mechanistically, metformin modulated the EGCG-activated Nrf2/HO-1 pathway through Sirtuin 1 (SIRT1)-dependent deacetylation of Nrf2. Moreover, metformin upregulated SIRT1 expression partially through the NF-kB pathway. Comparatively, the combination of EGCG and metformin showed little impact on normal lung epithelial BEAS-2B cells. Based on our findings, metformin sensitized NSCLC cells to the EGCG treatment by suppressing the Nrf2/HO-1 signaling pathway.

The Nrf2/GCH1/BH4 Axis Ameliorates Radiation-Induced Skin Injury by Modulating the ROS Cascade.

Radiation-induced skin injury is a common side effect of radiotherapy and can limit the duration and dose of radiotherapy. Most early work focused on elimination of reactive oxygen species (ROS) after radiation; however, less is known about the mechanisms underlying amplification of ROS and consequent skin injury by radiation. 5,6,7,8-Tetrahydrobiopterin (BH4) is an essential cofactor for all nitric oxide synthases. Inadequate availability of BH4 leads to uncoupling of nitric oxide synthases and production of highly oxidative radicals. In this study, we demonstrated that radiation disrupted BH4, which resulted in nitric oxide synthases uncoupling and augmented radiation-induced ROS. Overexpression of GTP cyclohydrolase I (GCH1), the rate-limiting enzyme for BH4 synthesis, restored cellular BH4 levels and nitric oxide production and decreased radiation-induced ROS. GCH1 also protected skin cells and rat skins against radiation-induced damage. We found that GCH1 was regulated by NF-E2-related factor 2, a key mediator of the cellular antioxidant response. Importantly, we identified GCH1 as a key effector for NF-E2-related factor 2-mediated protection against radiation-induced skin injury by inhibiting ROS production. Taken together, the findings of this study illustrate the key role of the NF-E2-related factor 2/GCH1/BH4 axis during radiation-induced skin damage.

REV3L modulates cisplatin sensitivity of non-small cell lung cancer H1299 cells.

Lung cancer remains the leading cause of cancer-related mortality worldwide and non-small cell lung cancer (NSCLC) accounts for approximately 80-85% of all cases of lung cancer. Cisplatin plays a significant role in the management of human lung cancer. Translesion DNA synthesis (TLS) is involved in DNA damage repair. DNA polymerase Î¶ (Pol Î¶) is able to mediate the DNA replication bypass of DNA damage, which is suggested to be involved in chemoresistance. REV3L is the catalytic subunit of Pol Î¶. Due to its critical role in translesion DNA synthesis, whether REV3L modulates cisplatin response in NSCLC cells remains unknown. In this study, REV3L overexpression and silencing H1299 cell lines were established. The reports showed that cisplatin induced the expression of REV3L by recruiting Sp1 to its promoter. Similar results were obtained when the ability of the cells to express luciferase from a platinated plasmid was measured. Co-transfection of the reporter with the REV3L overexpression vector or REV3L plus REV7L significantly enhanced the reporter activity. Nuclear condensation and fragmentation of shRNA-REV3L H1299 cells were more pronounced than shRNA-NC H1299 cells after cisplatin exposure, indicating that REV3L overexpression abolished cisplatin-induced DNA damage. Moreover, a forced expression of REV3L conferred the resistance of H1299 cells to cisplatin, whereas the knockdown of REV3L sensitized cisplatin efficacy in H1299 cells. Taken together, we demonstrated that inhibition of REV3L sensitized lung cancer H1299 cells to cisplatin treatment. Thus, REV3L may be a novel target for the chemotherapy of NSCLC.

Warburg meets non-coding RNAs: the emerging role of ncRNA in regulating the glucose metabolism of cancer cells.

Unlike normal differentiated cells, cancer cells primarily rely on glycolysis to generate energy needed for cellular processes even in normoxia conditions. This phenomenon is called aerobic glycolysis or "the Warburg effect." Aerobic glycolysis is inefficient to generate ATP, but the advantages it confers to cancer cells remain to be fully explained. Several oncogenic signaling pathways, interplaying with enzymes and kinases involved in glucose metabolism, participate in the switch from oxidative phosphorylation (OXPHOS) to aerobic glycolysis. Non-coding RNAs (ncRNAs) are a family of functional RNA molecules that are not further translated into proteins, which exert regulatatory roles in gene transcription and translation. ncRNAs, especially miRNAs and long non-coding RNAs (lncRNAs), may also have great effect on glucose metabolism by targeting not only glycolysis enzymes directly but also oncogenic signaling pathways indirectly. A better understanding of the Warburg effect and the regulatory role of ncRNAs in cancer glucose metabolism may contribute to the treatment of cancers.