Radiosensitization effect of quinoline-indole-schiff base derivative 10E on non-small cell lung cancer cells in vitro and in tumor xenografts.

Radioresistance is an inevitable obstacle in the clinical treatment of inoperable patients with non-small cell lung cancer (NSCLC). Combining treatment with radiosensitizers may improve the efficacy of radiotherapy. Previously, the quinoline derivative 10E as new exporter of Nur77 has shown superior antitumor activity in hepatocellular carcinoma. Here, we aimed to investigate the radiosensitizing activity and acting mechanisms of 10E. In vitro, A549 and H460 cells were treated with control, ionizing radiation (IR), 10E, and 10E + IR. Cell viability, apoptosis, and cycle were examined using CCK-8 and flow cytometry assays. Protein expression and localization were examined using western blotting and immunofluorescence. Tumor xenograft models were established to evaluate the radiosensitizing effect of 10E in vivo. 10E significantly inhibited cell proliferation and increased their radiosensitivity while reducing level of p-BCRA1, p-DNA-PKs, and 53BP1 involved in the DNA damage repair pathway, indicating that its radiosensitizing activity is closely associated with repressing DNA damage repair. A549 cells showed low level of Nur77 and a low response to IR but 10E-treated A549 cells showed high level of Nur77 indicating that Nur77 is a core radiosensitivity factor and 10E restores the expression of Nur77. Nur77 and Ku80 extranuclear co-localization in the 10E-treated A549 cells suggested that 10E-modulated Nur77 nuclear exportation inhibits DNA damage repair pathways and increases IR-triggered apoptosis. The combination of 10E and IR significantly inhibits tumor growth in a tumor xenograft model. Our findings suggest that 10E acts as a radiosensitizer and that combining 10E with radiotherapy may be a potential strategy for NSCLC treatment.

Genomic and functional impact of Trp53 inactivation in JAK2V617F myeloproliferative neoplasms.

Classical myeloproliferative neoplasms (MPNs) are characterized by the proliferation of myeloid cells and the risk of transformation into myelofibrosis or acute myeloid leukemia (AML) and TP53 mutations in MPN patients are linked to AML. However, JAK2V617F has been reported to impact the TP53 response to DNA damage, suggesting potential overlapping role of TP53 inactivation in MPN. We established a mouse model showing that JAK2V617F/Vav-Cre/Trp53-/- mice displayed a similar phenotype to JAK2V617F/Vav-Cre mice, but their proliferation was outcompeted in competitive grafts. RNA-Seq revealed that half of the genes affected by JAK2V617F were affected by p53-inactivation, including the interferon pathway. To validate this finding, mice were repopulated with a mixture of wild-type and JAK2V617F (or JAK2V617F/Vav-Cre/Trp53-/-) cells and treated with pegylated interferonα. JAK2V617F-reconstituted mice entered complete hematological remission, while JAK2V617F/Vav-Cre /Trp53-/--reconstituted mice did not, confirming that p53 loss induced interferon-α resistance. KEGG and Gene Ontology analyses of common deregulated genes showed that these genes were mainly implicated in cytokine response, proliferation, and leukemia evolution, illustrating that in this mouse model, the development of MPN is not affected by TP53 inactivation. Taken together, our results show that many genetic modifications induced by JAK2V617F are influenced by TP53, the MPN phenotype may not be. Trp53 loss alone is insufficient to induce rapid leukemic transformation in steady-state hematopoiesis in JAK2V617F MPN, and Trp53 loss may contribute to interferon resistance in MPN.

Tetrahedral DNA loaded siCCR2 restrains M1 macrophage polarization to ameliorate pulmonary fibrosis in chemoradiation-induced murine model.

Idiopathic pulmonary fibrosis (IPF) is a chronic lethal disease in the absence of demonstrated efficacy for preventing progression. Although macrophage-mediated alveolitis is determined to participate in myofibrotic transition during disease development, the paradigm of continuous macrophage polarization is still under-explored due to lack of proper animal models. Here, by integrating 2.5 U/kg intratracheal Bleomycin administration and 10 Gy thorax irradiation at day 7, we generated a murine model with continuous alveolitis-mediated fibrosis, which mimics most of the clinical features of our involved IPF patients. In combination with data from scRNA-seq of patients and a murine IPF model, a decisive role of CCL2/CCR2 axis in driving M1 macrophage polarization was revealed, and M1 macrophage was further confirmed to boost alveolitis in leading myofibroblast activation. Multiple sticky-end tetrahedral framework nucleic acids conjunct with quadruple ccr2-siRNA (FNA-siCCR2) was synthesized in targeting M1 macrophages. FNA-siCCR2 successfully blocked macrophage accumulation in pulmonary parenchyma of the IPF murine model, thus preventing myofibroblast activation and leading to the disease remitting. Overall, our studies lay the groundwork to develop a novel IPF murine model, reveal M1 macrophages as potential therapeutic targets, and establish new treatment strategy by using FNA-siCCR2, which are highly relevant to clinical scenarios and translational research in the field of IPF.

Granzyme B + CD8 + T cells with terminal differentiated effector signature determine multiple sclerosis progression.

BACKGROUND: Multiple sclerosis (MS) leads to demyelination and neurodegeneration with autoimmune responses in central nervous system. Patients begin with a relapsing-remitting (RR) course, and more than 80% of them may advance to secondary progressive MS (SPMS), which is characteristic for the gradual decline of neurological functions without demonstrated treating method to prevent. This study aims to investigate the contribution of peripheral CD8 + T cells during the conversion from RRMS to SPMS, as well as reveal potential diagnostic signature in distinguishing SPMS. METHODS: Single-cell RNA sequencing was employed to reveal the heterogeneity of CD8 + T cells between SPMS and RRMS. In addition, flow cytometry was used to further characterized CD8 + T cell dynamic changes in patients. T cell receptor sequencing was performed to detect the clonal expansion of MS. Using Tbx21 siRNA, T-bet was confirmed to manipulate GzmB expression. The correlation between GzmB + CD8 + T cell subsets and clinical characteristics of MS and their potential diagnostic value for SPMS were evaluated by generalized linear regression models and receiver operating characteristic (ROC) curve respectively. RESULTS: Other than diminished naïve CD8 + T cell, elevating of activated CD8 + T cell subsets were observed in SPMS patients. Meanwhile, this aberrant amplified peripheral CD8 + T cells not only exhibited terminal differentiated effector (EMRA) phenotype with GzmB expression, but also possessed distinct trajectory from clonal expansion. In addition, T-bet acted as a key transcriptional factor that elicited GzmB expression in CD8 + TEMRA cells of patients with SPMS. Finally, the expression of GzmB in CD8 + T cells was positively correlated with disability and progression of MS, and could effectively distinguish SPMS from RRMS with a high accuracy. CONCLUSIONS: Our study mapped peripheral immune cells of RRMS and SPMS patients and provided an evidence for the involvement of GzmB + CD8 + TEMRA cells in the progression of MS, which could be used as a diagnostic biomarker for distinguishing SPMS from RRMS.

A propensity score-matched study on the short-term outcome of ruptured blood blister-like aneurysm treated by microsurgery or endovascular surgery: a single-center study of 155 cases.

Treating blood blister-like aneurysms (BBA) is a major neurosurgical challenge. Whether endovascular repair serves as a better strategy than microsurgery remains controversial. We aim to perform a propensity score-matched (PSM) retrospective study to analyze the short-term outcome in BBA patients who received microsurgery and endovascular treatment. One hundred fifty-five eligible patients with internal carotid artery BBA were retrospectively collected with demographic and angiographic baseline in a single center. Three-month outcome and adverse events were set as outcome endpoints. PSM was used to match the microsurgery and endovascular group. Matching effect was evaluated by distribution variation analysis and love plot. The outcome of neurosurgery and endovascular treatment was then compared before and after PSM. Better WFNS levels (p = .017) and modified Fisher grade (p = .027) were noted in endovascular group before matching. Other baseline including angiographic features were comparable between two groups. Before matching, the 3-month outcome of endovascular repair surgery was more favorable than microsurgery (p < .0001). The occurrence of adverse events was also higher in the microsurgery group (p = .0079). In PSM-adjusted groups, the superior outcome effect of endovascular treatment still existed but with a reduced significance (p = .004). Similar trend was also observed in the adverse event rate (p = .038). Fatality rate was comparable between two adjusted groups regardless of PSM adjustment. Endovascular surgery of BBAs exhibits overall more favorable short-term outcome regardless of PSM matching. Microsurgery does not cause a higher fatality rate, hence it could be considered a salvage plan for those high-grade BBA patients.

AMPK pathway is implicated in low level lead-induced pubertal testicular damage via disordered glycolysis.

Lead (Pb) is a common environmental pollutant. It has been demonstrated that long-term exposure to Pb at environmental levels may cause severe and irreversible damage to the male reproductive system. Of note, the impairments may originate from environmental Pb exposure at puberty. However, the underlying mechanisms remain unclear. In this study, we administrated male ICR mice with 200 mg/L Pb through the drinking water for 30-, 60-, 90-day from postnatal day 28. RNA sequencing was performed in the control group and the 90-day Pb exposure group. It was found that Pb exposure induced testicular damage, increased oxidative stress levels and poor sperm quality. Bioinformatic analysis displayed 199 genes up-regulated (such as GLUT1 and MCT4 genes) and 156 genes down-regulated (such as GLUT3, PFK1, LDH, CD147 and AMPK genes) in the Pb exposure group compared to the control group. Gene ontology (GO) terms enrichment analysis showed differentially expressed genes (DEGs) are involved in the protein catabolic, cellular catabolic and triglyceride catabolic processes. KEGG pathways enrichment analysis indicated glycerolipid metabolism and AMPK signaling were significantly enriched. Furthermore, experimental verification showed that Pb exposure induces energy dysmetabolism and decreases glycolysis products in mice testicular tissue. The AMPK signaling pathway was found to be deactivated after Pb exposure. The GLUT1, GLUT3, PFK1 and LDH proteins, which play a critical role in the cell glycolysis process, also were decreased. Besides, the expression of CD147 was decreased and the location of CD147 was altered upon Pb exposure. Together, these findings indicated the implication of the AMPK signaling pathway in Pb exposure induced pubertal testicular damage and poor sperm quality by inhibiting cell glycolysis and disordering lactate transportation in testicular cells.

Marginal Zinc Deficiency in Mice Increased the Number of Abnormal Sperm and Altered the Expression Level of Spermatogenesis-Related Genes.

Marginal zinc deficiency is more common than severe zinc deficiency, and the effect of marginal zinc deficiency on male reproduction is unknown. This study investigated the effect of marginal zinc deficiency on spermatogenesis and its mechanism. Male ICR mice were fed normal zinc (30 mg/kg) and marginal zinc deficiency (10 mg/kg) diets for 35 days. Zinc-dependent proteins and enzymes were measured as biomarkers of zinc levels in the body. Metallothionein and Cu-Zn SOD levels in the control group were higher than those in the marginal zinc deficiency group. Hematoxylin and eosin staining showed that the marginal zinc deficiency diet caused histopathological changes in the testis and destruction of the sperm head under scanning electron microscopy. Sperm parameters and sex hormone levels were also affected by marginal zinc deficiency. In addition, marginal zinc deficiency led to alter expression level of several important spermatogenesis-related genes in the epididymis and testes. These results indicate that although zinc intake in marginal zinc deficiency is close to the recommended reference value, low zinc intake interferes with the expression of genes related to spermatogenesis and may lead to sperm abnormalities in mice.

Zinc-Enriched Yeast May Improve Spermatogenesis by Regulating Steroid Production and Antioxidant Levels in Mice.

Zinc (Zn) is an essential nutrient for the human body. This nutrient is involved in numerous physiological functions and plays an important role in spermatogenesis. Zn-enriched yeast (ZnY) is considered a Zn supplement with high bioavailability and is widely used as a functional food. However, the effect of ZnY on male reproductive function remains unclear. This study aimed to investigate the beneficial effects of ZnY on the treatment of male spermatogenesis disorders. The spermatogenic dysfunctional mice were established by using cyclophosphamide (CP). CP was administered in saline at a dose of 50 mg/kg bw/day for 5 days by intraperitoneal injection (i.p.). Then, ZnY was orally supplemented at the dose levels of 2, 4, and 8 mg Zn/kg bw/day for 30 days. CP significantly decreased the sperm density and viability, testicular marker enzymes, serum testosterone, follicular stimulating hormone (FSH), and luteinizing hormone (LH). ZnY supplementation significantly improved these sperm parameters and hormone levels. Additionally, ZnY decreased the CP-induced lipid peroxidation and increased the glutathione levels. Moreover, ZnY increased the gene expression of anti-apoptotic proteins and steroid synthetase in mouse testes. The low-dose ZnY supplementation has a better effect on improving spermatogenesis, while the other two groups are less beneficial roles possibly due to excessive Zn intake. The present results suggest that appropriate ZnY can act as an accessory factor to improve steroid production and antioxidant levels in spermatogenic dysfunction mice.

Improvement roles of zinc supplementation in low dose lead induced testicular damage and glycolytic inhibition in mice.

Lead (Pb) is a toxic metal that affects the male reproductive system. This study aimed to investigate the effects of zinc (Zn) intake between recommended dietary allowances (RDAs) and tolerable upper intake levels (ULs) in preventing male testis damage induced by low-dose Pb. Forty-five mice were randomly divided into control, Pb, and Pb + Zn groups. They were given distilled water ad libitum with 0, 200 mg/L Pb2+, or 15 mg/L Zn2+ mixed with 200 mg/L Pb2+ for 90 consecutive days. The Zn levels in the blood and testis of the Pb group were significantly lower than those of the control group. The Pb levels in the blood and testis of the Pb + Zn group were significantly lower than those of the Pb group. Additionally, a significant decrease in sperm density and viability, with a significant increase in sperm abnormality rate and DNA fragmentation index, was observed in the Pb group. Zn supplementation significantly improved the above sperm parameters. Moreover, Zn supplementation decreased low-dose Pb-induced lipid peroxidation and increased glutathione, total superoxide dismutase (SOD), and copper/Zn-SOD levels. Furthermore, Zn treatment improved glycolysis products and lactate transporters in Pb-treated mouse testes. Our findings suggest that Zn intake between RDAs and UL can act as a therapeutic agent in protecting against the reproductive impairments associated with Pb exposure.

Genome-wide identification and functional analysis of long non-coding RNAs and mRNAs in male mice testes at the onset of puberty after low dose lead exposure.

Many researchers have studied the relationship between lead (Pb) and testis injury, but the underlying mechanisms are still unknown. The participation of long non-coding RNAs (lncRNAs) in biological processes has been proposed. To comprehensively gain insight into the molecular toxicity of Pb, expression patterns are analysed through RNA sequencing (RNA-seq) in male mice treated with 200 mg/L of Pb through the drinking water for 90 days at the onset of puberty. A total of 614 differentially expressed (DE) lncRNAs were included (p ≤ 0.05 and fold change ≥2), of which 288 were up-regulated, and 326 were down-regulated. A total of 2295 DE mRNAs (p ≤ 0.05 and fold change ≥2), including 1202 up-regulated and 1093 down-regulated ones, were found in the testes of Pb-exposed group. Functional analysis results showed that several lncRNAs might be implicated in the bio-pathway of mitogen-activated protein kinase (MAPK) signaling pathway. Finally, seven pairs of lncRNA-mRNA co-expression were established in mice testes and confirmed by RT-qPCR. Moreover, the DE genes were also altered in Sertoli cells. Therefore, our research might be helpful for future exploring the effects of Pb exposure on lncRNA in testis, as well as its function.

POLE and Mismatch Repair Status, Checkpoint Proteins and Tumor-Infiltrating Lymphocytes in Combination, and Tumor Differentiation: Identify Endometrial Cancers for Immunotherapy.

OBJECTIVE: Although Polymerase-epsilon (POLE)-mutated and mismatch repair (MMR)-deficient endometrial cancers (ECs) are considered as promising candidates for anti-PD-1/PD-L1 therapy, selecting only these patients may exclude other patients who could potentially respond to this treatment strategy, highlighting the need of additional biomarkers for better patient selection. This study aims to evaluate potential predictive biomarkers for anti-PD-1/PD-L1 therapy in addition to POLE mutation (POLEm) and MMR deficiency (MMRd). METHODS: We performed next generation sequencing for POLE from 202 ECs, and immunohistochemistry for MLH1, MSH2, MSH6, PMS2, CD3, CD8, PD-1 and PD-L1 on full-section slides from these ECs. We assessed the association of POLEm and MMRd with clinicopathologic features, expression of check point proteins, and density of tumor-infiltrating lymphocytes (TILs). Prognostic impact of these immune markers was also evaluated. RESULTS: POLEm, MMRd and high-grade tumors exhibited elevated level of TILs. Increased expression of PD-1 and PD-L1 was observed in MMRd and high-grade ECs. A subgroup of MMR proficient ECs also harbored increased density of TILs, and positive expression of PD-1 and PD-L1. In addition, negative expression of checkpoint proteins and high density of TILs in combination was associated with good prognosis. CONCLUSIONS: Candidates for PD-1 blockade may extend beyond POLEm and MMRd ECs, additional factors such as tumor grade, and combination of TILs levels and expression of checkpoint proteins may need to be considered for better patient selection.

Visualization and bibliometric analysis of cAMP signaling system research trends and hotspots in cancer.

Cyclic adenosine monophosphate (cAMP) is an essential second messenger that widely distributed among prokaryotic and eukaryotic organisms. cAMP can regulate various biological processes, including cell proliferation, differentiation, apoptosis and immune functions. Any dysregulation or alteration of cAMP signaling may cause cell metabolic disorder, immune dysfunction and lead to disease or cancer. This study aimed to conduct a scientometric analysis of cAMP signaling system in cancer field, and explored the research trend, hotspots and frontiers from the past decade. Relevant literatures published from 2009 to 2019 were collected in the Web of Science Core Collection database. EndNote X9 was used to remove duplicate articles, and irrelevant articles were manually filtered. Bibliometric analyses were completed by CiteSpace V. A total of 4306 articles were included in this study. The number of related literatures published each year is gradually increasing. Most of them belong to "Biochemistry & Molecular Biology", "Oncology", "Cell Biology", "Pharmacology & Pharmacy" and "Endocrinology & Metabolism" areas. In the past decade, USA, China, and Japan contributed the most to the research of cAMP signaling system in cancer. The frontiers and hotspots of cAMP signaling pathway system related to cancer fields mainly focused on cancer cell apoptosis, metastasis, and multiple tumors occurrence in patients with Carney complex. Intervention of the cAMP metabolic pathway may be a potential and promising therapeutic strategy for controlling clinical cancer and tumor diseases.

PbAc Triggers Oxidation and Apoptosis via the PKA Pathway in NRK-52E Cells.

This study aimed to investigate the mechanism of the lead exposure-induced oxidative stress and apoptosis of renal tubular epithelial cells. We explored the effects of lead acetate (PbAc) on the oxidation and apoptosis of renal proximal tubular cells (NRK-52E) through in vitro experiments. Results showed that PbAc induced dose-dependent reactive oxygen species (ROS) accumulation in NRK-52E cells, and the activities of superoxide dismutase (SOD) and glutathione (GSH) decreased, whereas the malondialdehyde (MDA) content increased. Under the exposure of 40 and 80 µM PbAc, the mRNA level of B cell lymphoma-2 (Bcl-2) in the cells decreased, the mRNA levels of Bcl-2-associated X protein (Bax) and caspase-3 increased, and apoptosis was obvious. Furthermore, the nicotinamide adenine dinucleotide phosphate oxidase 4 (Nox4) activity was enhanced by PbAc in a dose-dependent manner. The mRNA levels of protein kinase A (PKA) were upregulated by PbAc. H-89, a PKA inhibitor, suppressed PKA activation, ROS accumulation, and Nox4 activity in NRK-52E cells. Our results indicated that PbAc potentially stimulated oxidative stress and apoptosis in NRK-52E cells by increasing Nox4-dependent ROS production via the PKA signaling pathway.

Residential Radon and Histological Types of Lung Cancer: A Meta-Analysis of Case‒Control Studies.

Epidemiological studies on residential radon exposure and the risk of histological types of lung cancer have yielded inconsistent results. We conducted a meta-analysis on this topic and updated previous related meta-analyses. We searched the databases of Cochrane Library, Embase, PubMed, Web of Science and Chinese National Knowledge Infrastructure for papers published up to 13 November 2018. The pooled odds ratio (OR) and 95% confidence interval (CI) were calculated using fixed and random effects models. Subgroup and dose‒response analyses were also conducted. This study was registered with PROSPERO (No. CRD42019127761). A total of 28 studies, which included 13,748 lung cancer cases and 23,112 controls, were used for this meta-analysis. The pooled OR indicated that the highest residential radon exposure was significantly associated with an increased risk of lung cancer (OR = 1.48, 95% CI = 1.26-1.73). All histological types of lung cancer were associated with residential radon. Strongest association with small-cell lung carcinoma (OR = 2.03, 95% CI = 1.52-2.71) was found, followed by adenocarcinoma (OR = 1.58, 95% CI = 1.31-1.91), other histological types (OR = 1.54, 95% CI = 1.11-2.15) and squamous cell carcinoma (OR = 1.43, 95% CI = 1.18-1.74). With increasing residential radon levels per 100 Bq/m3, the risk of lung cancer, small-cell lung carcinoma and adenocarcinoma increased by 11%, 19% and 13%, respectively. This meta-analysis provides new evidence for a potential relationship between residential radon and all histological types of lung cancer.

Lead-induced oxidative damage in rats/mice: A meta-analysis.

BACKGROUND: Lead (Pb) is ubiquitous in the environment and is an environmental genotoxic metal. Pb accumulation in the body could cause the oxidative stress. OBJECTIVE: This meta-analysis aimed to perform a systematic evaluation of the extent of oxidative damage in rats/mice induced by lead. METHODS: All relevant articles in English or Chinese were retrieved from Embase, PubMed, Web of Science, Medline, China National Knowledge Infrastructure, and Chinese Biological Medicine databases from their inception date until July 22, 2018. RESULTS: A total of 108 eligible articles were included in this study. The indicators of oxidative stress included malondialdehyde (MDA), glutathione disulfide (GSSG), reactive oxygen species (ROS), hydrogen peroxide (H2O2), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), reduced glutathione (GSH), superoxide dismutase (SOD), and glutathione-s-transferase (GST). The meta-analysis showed that lead significantly increased oxidants levels, such as MDA, GSSG, ROS, and H2O2 (P < 0.05), and significantly reduced the level of antioxidants, such as CAT, GPx, GR, GSH, SOD, and GST (P < 0.05). The intraperitoneal mode was more effective than water drinking mode in reducing the levels of CAT, GPx, GSH, and SOD (P < 0.05). Other factors that influenced the overall oxidative stress, including species of animals, type of tissues, and intervention dosage and time, were comprehensively evaluated. CONCLUSION: The results of meta-analysis indicated that mice were more sensitive to lead than rats, and intraperitoneal mode was an effective intervention mean. High doses and long periods of lead treatment can cause serious oxidative damage. Moreover, testicular was more vulnerable to lead than other tissues. These results provided scientific evidence for preventing and treating lead toxicity.