Integration of RNA-seq and ATAC-seq analyzes the effect of low dose neutron-γ radiation on gene expression of lymphocytes from oilfield logging workers.

Objective: Although radiation workers are exposed to much lower doses of neutron-γ rays than those suffered in nuclear explosions and accidents, it does not mean that their health is not affected by radiation. Lower doses of radiation do not always cause morphological aberrations in chromosomes, so more sophisticated tests must be sought to specific alterations in the exposed cells. Our goal was to characterize the specific gene expression in lymphocytes from logging workers who were continuously exposed to low doses of neutron-γ radiation. We hypothesized that the combination of cell type-specific transcriptomes and open chromatin profiles would identify lymphocyte-specific gene alterations induced by long-term radiation with low-dose neutron-γ-rays and discover new regulatory pathways and transcriptional regulatory elements. Methods: Lymphocytes were extracted from workers who have been occupationally exposed to neutron-γ and workers unexposed to radiation in the same company. mRNA-seq and ATAC-seq (Assay for Transposase-Accessible Chromatin with high-throughput sequencing) were performed, followed integrative analysis to identify specific gene regulatory regions induced by neutron-γ radiation. A qPCR assay was then performed to verify the downregulation of RNA coding for ribosomal proteins and flow cytometry was used to detect ribosomal protein expression and cell cycle alterations. Results: We identified transcripts that were specifically induced by neutron-γ radiation and discovered differential open chromatin regions that correlated with these gene activation patterns. Notably, we observed a downward trend in the expression of both differentially expressed genes and open chromatin peaks. Our most significant finding was that the differential peak upregulated in ATAC-seq, while the differential gene was downregulated in the ribosome pathway. We confirmed that neutron-γ radiation leads to transcriptional inhibition by analyzing the most enriched promoters, examining RPS18 and RPS27A expression by qPCR, and analyzing protein-protein interactions of the differential genes. Ribosomal protein expression and cell cycle were also affected by neutron-γ as detected by flow cytometry. Conclusion: We have comprehensively analyzed the genetic landscape of human lymphocytes based on chromatin accessibility and transcript levels, enabling the identification of novel neutron-γ induced signature genes not previously known. By comparing fine-mapping of open chromatin and RNA reads, we have determined that neutron-γ specifically leads to downregulation of genes in the ribosome pathway, with pseudogenes potentially playing a crucial role.

RMI1 facilitates repair of ionizing radiation-induced DNA damage and maintenance of genomic stability.

Ionizing radiation (IR) causes a wide variety of DNA lesions, of which DNA double-stranded breaks (DSBs) are the most deleterious. Homologous recombination (HR) is a crucial route responsible for repairing DSBs. RecQ-mediated genome instability protein 1 (RMI1) is a member of an evolutionarily conserved Bloom syndrome complex, which prevents and resolves aberrant recombination products during HR, thereby promoting genome stability. However, little is known about the role of RMI1 in regulating the cellular response to IR. This study aimed to understand the cellular functions and molecular mechanisms by which RMI1 maintains genomic stability after IR exposure. Here, we showed IR upregulated the RMI1 protein level and induced RMI1 relocation to the DNA damage sites. We also demonstrated that the loss of RMI1 in cells resulted in enhanced levels of DNA damage, sustained cell cycle arrest, and impaired HR repair after IR, leading to reduced cell viability and elevated genome instability. Taken together, our results highlighted the direct roles of RMI1 in response to DNA damage induced by IR and implied that RMI1 might be a new genome safeguard molecule to radiation-induced damage.

Assessment of radiation doses and DNA damage in pediatric patients undergoing interventional procedures for vascular anomalies.

Interventional procedures (IPs) have been widely used to treat vascular anomalies (VA) in recent years. However, patients are exposed to low-dose X-ray ionizing radiation (IR) during these fluoroscopy-guided IPs. We collected clinical information and IR doses during IPs and measured biomarkers including γ-H2AX, chromosome aberrations (CA), and micronuclei (MN), which underpin radiation-induced DNA damage, from 74 pediatric patients before and after IPs. For the 74 children, the range of dose-area product (DAP) values was from 1.2 to 1754.6 Gy∙cm2, with a median value of 27.1 Gy∙cm2. DAP values were significantly higher in children with lesions in the head and neck than in the limbs and trunk; the age and weight of children revealed a strong positive correlation with DAP values. The treated patients as a group demonstrated an increase in all three endpoints relative to baseline following IPs. Children with vascular tumors have a higher risk of dicentric chromosome + centric ring (dic+r) and cytokinesis-block micronucleus (CBMN) after IPs than children with vascular malformations. The younger the patient, the greater the risk of CA after IPs. Moreover, rogue cells (RCs) were found in five children (approximately 10%) after IPs, and the rates of dic+r and CBMN were significantly higher than those of other children (Z = -3.576, p < 0.001). These results suggest that there may be some children with VA who are particularly sensitive to IR, but more data and more in-depth experiments will be needed to verify this in the future.

Identification and Bioinformatic Assessment of circRNA Expression After RMI1 Knockdown and Ionizing Radiation Exposure.

RecQ-mediated genome instability protein 1 (RMI1) is an important component of the BLM-Topo IIIα-RMI1-RMI2 complex and plays a critical role in maintaining genome stability. However, the cellular functions of RMI1 in response to ionizing radiation (IR) are poorly understood. In this study, we found that RMI1 knockdown led to enhanced radiosensitivity and apoptosis after irradiation. To analyze the effect of RMI1 knockdown on the expression of circular RNAs (circRNAs), we performed high-throughput RNA sequencing on four groups of human embryonic kidney (HEK) 293T cells: control cells and RMI1 knockdown cells with or without IR exposure. A total of 179 and 160 differentially expressed circRNAs (DE-circRNAs) were identified under RMI1 knockdown without and with exposure to IR, respectively. The Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed that these DE-circRNAs were involved in a variety of functions and signal pathways, including histone H3-K36 methylation, nuclear pore organization, mRNA destabilization, the mismatch repair pathway, and the apoptotic signaling pathway. Overall, our results indicate that RMI1 plays a crucial role in the response to IR and, more generally, that circRNAs are important in the regulatory mechanism of the radiation response.

Anxiety and Depression Among Imaging Doctors in Post-COVID-19 Period.

To investigate the mental state of medical imaging staff in Shandong Province, China, who have been on the forefront of the COVID-19 epidemic during its late stage in China. Questionnaires designed to assess anxiety and depression were administered on-location, and 5331 complete results were collected. SPSS software was used for statistical descriptions and analysis. Rates of anxiety disorders and depression among medical imaging workers in Shandong Province, China, were 6.1% and 6.5%, respectively, higher than those of anxiety and depression in Chinese residents before the epidemic. The outbreak in Xinjiang, China; virus mutation in Japan; and spread of the epidemic due to occupational errors were the primary reported causes of anxiety and depression among image workers. Medical imaging workers showed evidence of psychological abnormalities during the late stage of the epidemic in China.

Exposure Doses to Technologists Working in 7 PET/CT Departments.

OBJECTIVE: The aim of this study was to measure occupational exposure doses of technologists who dispense and inject radiopharmaceuticals in 7 positron emission tomography/computed tomography (PET/CT) departments. This was done with the goal to help improving protective designs in PET departments and/or establishing national protection standards. METHOD: Common LiF thermoluminescence dosimeters (TLDs) were placed on the chest and necklace of the technologists to monitor whole-body and thyroid doses, respectively. Ring TLDs were also worn on both index fingers to measure individual hand doses. All TLDs were assembled and measured once every 3 months for a total of 12 months. Additionally, we measured and compared the dose of TLDs attached to both the inside and the outside of the technologist's lead coat. RESULTS: Technologists received relatively high exposures, which accounted for 64% to 94% of the collective dose in their respective departments. Their thyroid doses ranged from 1.2 to 1.7 mSv/a; some technologists' hand doses exceeded 500 mSv/a. Use of a lead coat reduced the average dose by 8%. CONCLUSION: Technologists working in PET/CT departments were the main population exposed to radiation. This work underscores the need for enhanced protective measures for these workers to better reduce their exposure, particularly for their hands.

Assessment of Genomic Instability in Medical Workers Exposed to Chronic Low-Dose X-Rays in Northern China.

The increasing use of ionizing radiation (IR) in medical diagnosis and treatment has caused considerable concern regarding the effects of occupational exposure on human health. Despite this concern, little information is available regarding possible effects and the mechanism behind chronic low-dose irradiation. The present study assessed potential genomic damage in workers occupationally exposed to low-dose X-rays. A variety of analyses were conducted, including assessing the level of DNA damage and chromosomal aberrations (CA) as well as cytokinesis-block micronucleus (CBMN) assay, gene expression profiling, and antioxidant level determination. Here, we report that the level of DNA damage, CA, and CBMN were all significantly increased. Moreover, the gene expression and antioxidant activities were changed in the peripheral blood of men exposed to low-dose X-rays. Collectively, our findings indicated a strong correlation between genomic instability and duration of low-dose IR exposure. Our data also revealed the DNA damage repair and antioxidative mechanisms which could result in the observed genomic instability in health-care workers exposed to chronic low-dose IR.

Antioxidant Activity Evaluation of Dietary Flavonoid Hyperoside Using Saccharomyces Cerevisiae as a Model.

Oxidative stress leads to various diseases, including diabetes, cardiovascular diseases, neurodegenerative diseases, and even cancer. The dietary flavonol glycoside, hyperoside (quercetin-3-O-galactoside), exerts health benefits by preventing oxidative damage. To further understand its antioxidative defence mechanisms, we systemically investigated the regulation of hyperoside on oxidative damage induced by hydrogen peroxide, carbon tetrachloride, and cadmium in Saccharomyces cerevisiae. Hyperoside significantly increased cell viability, decreased lipid peroxidation, and lowered intracellular reactive oxygen species (ROS) levels in the wild-type strain (WT) and mutants gtt1∆ and gtt2∆. However, the strain with ctt1∆ showed variable cell viability and intracellular ROS-scavenging ability in response to the hyperoside treatment upon the stimulation of H2O2 and CCl4. In addition, hyperoside did not confer viability tolerance or intercellular ROS in CdSO4-induced stress to strains of sod1∆ and gsh1∆. The results suggest that the antioxidative reactions of hyperoside in S. cerevisiae depend on the intercellular ROS detoxification system.

RMI1 contributes to DNA repair and to the tolerance to camptothecin.

Maintenance of genome integrity is critical for faithful propagation of genetic information and the prevention of the mutagenesis induced by various DNA damage events. RecQ-mediated genome instability protein 1 (RMI1), together with Bloom syndrome protein and topoisomerase IIIα, form an evolutionarily conserved complex that is critical for the maintenance of genomic stability. Herein, we report that RMI1 depletion increases cell sensitivity to camptothecin treatment, as shown by an elevation of genotoxic stress-induced DNA double-strand breaks, a stronger activation of the DNA damage response, and a greater G2/M cell cycle delay. Our findings support that, upon DNA damage, RMI1 forms nuclear foci at the damaged regions, interacts with RAD51, and facilitates the recruitment of RAD51 to initiate homologous recombination. Our data reveal the importance of RMI1 in response to DNA double-strand breaks and shed light on the molecular mechanisms by which RMI1 contributes to maintain genome stability.-Fang, L., Sun, X., Wang, Y., Du, L., Ji, K., Wang, J., He, N., Liu, Y., Wang, Q., Zhai, H., Hao, J., Xu, C., Liu, Q. RMI1 contributes to DNA repair and to the tolerance to camptothecin.

Anti-proliferative and anti-invasive effects of garcinol from Garcinia indica on gallbladder carcinoma cells.

Garcinol, a natural histone acetyltransferase inhibitor, has been reported to exhibit significant anti-proliferative activity in various cancer cell types. However, no information is available about the anti-cancer effects of garcinol on gallbladder carcinoma cells (GBC). In this study, GBC cells (GBC-SD and NOZ) were treated by garcinol and subjected to Cell Counting Kit-8 (CCK-8), and GBC-SD cells were selected for further transwell chamber assay, quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis. Our results indicated that garcinol could significantly inhibit the growth of GBC cells in a dose- and time-dependent manner. It also inhibited the invasion of GBC-SD cells in a dose-dependent manner. Garcinol treatment decreased the activity of matrix metalloproteinase 2 (MMP2) and MMP9 by the downregulation of mRNA levels, and these two enzymes are critical to tumor invasion. Treatment with garcinol also decreased Stat3 and Akt activation in GBC-SD cells. Taken together, the effects of garcinol on GBC-SD cells may be associated with the suppression of Stat3 and Akt signaling pathways, which may contribute to inhibiting their downstream targets such as mRNA levels of MMP2 and MMP9.

Genomic instability in adult men involved in processing electronic waste in Northern China.

BACKGROUND: Managing and recycling electronic waste (e-waste), while useful and necessary, has resulted in significant contamination of several environments in China. The area around Tianjin, China has become one of the world's largest e-waste disposal centers, where electronics are processed by manually disassembly or burning, which can result in serious exposure of workers to a multitude of toxicants. OBJECTIVE: The present study assessed potential genomic damage in workers involved in recycling e-waste. METHODS: To detect cytogenetic and DNA damage, chromosomal aberrations (CA), cytokinesis blocking micronucleus (CBMN) and the comet assay were performed. Concentrations of some trace elements, markers of oxidative stress and polychlorinated biphenyls (PCBs) in whole blood or serum were measured, and relationships among the markers described above, age, and duration of exposure were analyzed. The profiles of expression of genes in lymphocytes in peripheral blood were assessed to determine the status of the regulation of genes involved in genome stability. RESULTS: Concentrations of 28 PCB congeners in the whole blood of the exposed group were significantly (P<0.001) greater than those in the control individuals. Frequency of CA (8.01%) and CBMN (26.3‰) in lymphocytes and the level of DNA damage in the lymphocytes and spermatozoa of the exposed men were also significantly (P<0.0001) greater than those of the controls. There were significant relationships between CA, CBMN, DNA damage and duration of exposure. Concentrations of malondialdehyde (MDA) and lead (Pb) in the blood serum were significantly greater, but activities of superoxide dismutase (SOD), glutathione (GSH) and concentrations of calcium (Ca) and magnesium (Mg) were lower in the serum of the exposed men. MDA, Pb, Ca and Mg were associated with the duration of exposure to handling e-waste. In males involved in handling of e-waste, there were 13 genes - ATM, ATR, ABL1, CHEK1, CHEK2, GADD45A, CDK7, GTSE1, OGG1, DDB1, PRKDC, XRCC1 and CCNH - for which expression of mRNA was up-regulated and 7 genes - BRCA1, GTF2H1, SEMA4A, MRE11A, MUTYH, PNKP and RAD50 - for which the expression of mRNA was down-regulated. CONCLUSIONS: A strong correlation between indicators of damage of DNA, which could result in instability of the genome, and duration of processing e-waste was observed. If proper procedures are not followed, there are significant risks to the health of the individuals involved in such activities.

Ginger Oleoresin Alleviated γ-Ray Irradiation-Induced Reactive Oxygen Species via the Nrf2 Protective Response in Human Mesenchymal Stem Cells.

Unplanned exposure to radiation can cause side effects on high-risk individuals; meanwhile, radiotherapies can also cause injury on normal cells and tissues surrounding the tumor. Besides the direct radiation damage, most of the ionizing radiation- (IR-) induced injuries were caused by generation of reactive oxygen species (ROS). Human mesenchymal stem cells (hMSCs), which possess self-renew and multilineage differentiation capabilities, are a critical population of cells to participate in the regeneration of IR-damaged tissues. Therefore, it is imperative to search effective radioprotectors for hMSCs. This study was to demonstrate whether natural source ginger oleoresin would mitigate IR-induced injuries in human mesenchymal stem cells (hMSCs). We demonstrated that ginger oleoresin could significantly reduce IR-induced cytotoxicity, ROS generation, and DNA strand breaks. In addition, the ROS-scavenging mechanism of ginger oleoresin was also investigated. The results showed that ginger oleoresin could induce the translocation of Nrf2 to cell nucleus and activate the expression of cytoprotective genes encoding for HO-1 and NQO-1. It suggests that ginger oleoresin has a potential role of being an effective antioxidant and radioprotective agent.

Knockdown of RMI1 impairs DNA repair under DNA replication stress.

RMI1 (RecQ-mediated genome instability protein 1) forms a conserved BTR complex with BLM, Topo IIIα, and RMI2, and its absence causes genome instability. It has been revealed that RMI1 localizes to nuclear foci with BLM and Topo IIIα in response to replication stress, and that RMI1 functions downstream of BLM in promoting replication elongation. However, the precise functions of RMI1 during replication stress are not completely understood. Here we report that RMI1 knockdown cells are hypersensitive to hydroxyurea (HU). Using comet assay, we show that RMI1 knockdown cells exhibit accumulation of broken DNAs after being released from HU treatment. Moreover, we demonstrate that RMI1 facilitates the recovery from activated checkpoint and resuming the cell cycle after replicative stress. Surprisingly, loss of RMI1 results in a failure of RAD51 loading onto DNA damage sites. These findings reveal the importance of RMI1 in response to replication stress, which could explain the molecular basis for its function in maintaining genome integrity.

Dietary Flavonoid Hyperoside Induces Apoptosis of Activated Human LX-2 Hepatic Stellate Cell by Suppressing Canonical NF-κB Signaling.

Hyperoside, an active compound found in plants of the genera Hypericum and Crataegus, is reported to exhibit antioxidant, anticancer, and anti-inflammatory activities. Induction of hepatic stellate cell (HSC) apoptosis is recognized as a promising strategy for attenuation of hepatic fibrosis. In this study, we investigated whether hyperoside treatment can exert antifibrotic effects in human LX-2 hepatic stellate cells. We found that hyperoside induced apoptosis in LX-2 cells and decreased levels of α-smooth muscle actin (α-SMA), type I collagen, and intracellular reactive oxygen species (ROS). Remarkably, hyperoside also inhibited the DNA-binding activity of the transcription factor NF-κB and altered expression levels of NF-κB-regulated genes related to apoptosis, including proapoptotic genes Bcl-Xs, DR4, Fas, and FasL and anti-apoptotic genes A20, c-IAP1, Bcl-X L , and RIP1. Our results suggest that hyperoside may have potential as a therapeutic agent for the treatment of liver fibrosis.

Anticancer activities of citrus peel polymethoxyflavones related to angiogenesis and others.

Citrus is a kind of common fruit and contains multiple beneficial nutrients for human beings. Flavonoids, as a class of plant secondary metabolites, exist in citrus fruits abundantly. Due to their broad range of pharmacological properties, citrus flavonoids have gained increased attention. Accumulative in vitro and in vivo studies indicate protective effects of polymethoxyflavones (PMFs) against the occurrence of cancer. PMFs inhibit carcinogenesis by mechanisms like blocking the metastasis cascade, inhibition of cancer cell mobility in circulatory systems, proapoptosis, and antiangiogenesis. This review systematically summarized anticarcinogenic effect of citrus flavonoids in cancer therapy, together with the underlying important molecular mechanisms, in purpose of further exploring more effective use of citrus peel flavonoids.