miR-584-5p / Ykt6 - mediated autophagy - lysosome - exosome pathway as a critical route affecting the toxic effects of lead on HK-2 cells.

Lead is a widespread environmental pollutant with serious adverse effects on human health, but the mechanism underlying its toxicity remains elusive. This study aimed to investigate the role of miR-584-5p / Ykt6 axis in the toxic effect of lead on HK-2 cells and the related mechanism. Our data suggested that lead exposure caused significant cytotoxicity, DNA and chromosome damage to HK-2 cells. Mechanistically, lead exposure down-regulated miR-584-5p and up-regulated Ykt6 expression, consequently, autophagosomal number and autophagic flux increased, lysosomal number and activity decreased, exosomal secretion increased. Interestingly, when miR-584-5p level was enhanced with mimic, autophagosomal number and autophagic flux decreased, lysosomal number and activity increased, ultimately, exosomal secretion was down-regulated, which resulted in significant aggravated toxic effects of lead. Further, directly blocking exosomal secretion with inhibitor GW4869 also resulted in exacerbated toxic effects of lead. Herein, we conclude that miR-584-5p / Ykt6 - mediated autophagy - lysosome - exosome pathway may be a critical route affecting the toxic effects of lead on HK-2 cells. We provide a novel insight into the mechanism underlying the toxicity of lead on human cells.

HUC-MSC-derived exosomes repaired the damage induced by hydroquinone to 16HBE cells via miR-221/PTEN pathway.

Mesenchymal stem cell - originated exosomes (MSC-exo) are promising non-cellular treatment agents for various diseases. The present study aimed to explore whether human umbilical cord MSC - originated exosomes (HUC-MSC-exo) have the function of protecting human cells (16HBE) against the damage caused by HQ and the related mechanism. HUC-MSC-exo was isolated with differential gradient ultracentrifugation method and characterized by using transmission electron microscope (TEM). 16HBE cells were used as the tool cells and co-cultured with HUC-MSC-exo. Confocal laser scanning microscope was employed to confirm the ingestion of HUC-MSC-exo by 16HBE. Cell proliferation, migration, oxidative stress, DNA and chromosome damages of 16HBE were analyzed under HQ stress, and the role of miR-221/PTEN axis was investigated. Our data showed that under HQ stress, different groups of cells exhibited significantly decreased proliferation and migration abilities, and significant oxidative stress, DNA and chromosome damage effects. HUC-MSC-exo could alleviate the cytotoxic, oxidative stress and genotoxic damage effects of HQ on 16HBE cells. Mechanistically, HQ exposure up-regulated the level of miR-221 and down-regulated PTEN, while HUC-MSC-exo could significantly reduce the level of miR-221 and promote PTEN expression, which was involved in alleviating the toxic effects of HQ on 16HBE cells. Our data indicates that HUC-MSC-exo can alleviate the oxidative stress, cytotoxic and genotoxic effects of HQ on 16HBE cells via miR-221/PTEN pathway, and it may be a promising agent for protecting against the toxicity of HQ.

[dvances in the diagnosis and treatment of necrozoospermia].

Necrozoospermia is a special type of asthenospermia, in which mass sperm death is commonly seen, with an incidence rate of 0.2%－0.4%. Studies on necrospermia are rarely reported. Its etiology is complicated, and its diagnosis and treatment are very difficult. This article focuses on the main etiological factors, pathophysiological mechanism, diagnostic methods and treatment strategies of necrospermia, aiming to provide some reference for andrologists and reproduction physicians, as well as a theoretical guidance for intracytoplasmic sperm injection (ICSI) in the treatment of the patients with necrospermia.

Lysosome-related exosome secretion mediated by miR-26b / Rab31 pathway was associated with the proliferation and migration of MCF-7 cells treated with BPA.

Bisphenol A (BPA), one of the typical environmental endocrine disruptors (EEDs), can promote the proliferation and migration of cancer cells, but the mechanism of which remains largely unclear. Exosome secretion plays an important role in the stress response of cells to environmental stimuli. This study was designed to explore whether exosome secretion was involved in the toxic effect of BPA on the proliferation and migration of MCF-7 cells, and the related mechanism. Our data shows that the IC50 value of MCF-7 exposure to BPA was about 65.82 µM. The exposure of MCF-7 to 10 µM BPA resulted in a decreased miR-26b expression and the activation of miR-26b/Rab-31 pathway, consequently, the number and activity of lysosomes decreased, the secretion of exosomes increased, cell proliferation and migration were enhanced obviously. Interestingly, miR-26b mimic up-regulated the number and activity of lysosomes via miR-26b/miR-31 pathway, exosome secretion was down-regulated, cell proliferation and migration decreased. Further, when GW4869 was used to directly inhibit the exosome secretion of MCF-7 treated with BPA, their proliferation and migration were down-regulated. Herein, we concluded that the stimulating effect of BPA on the proliferation and migration of MCF-7 cells was associated with the lysosome - related exosome secretion via miR-26b / Rab31 pathway.

Exploring the Causal Relationship Between Migraine and Insomnia Through Bidirectional Two-Sample Mendelian Randomization: A Bidirectional Causal Relationship.

Introduction: The intricate relationship between migraine and insomnia has been a subject of great interest due to its complex mechanisms. Despite extensive research, understanding the causal link between these conditions remains a challenge. Material and Methods: This study employs a bidirectional Mendelian randomization approach to investigate the causal relationship between migraine and insomnia. Risk loci for both conditions were derived from large-scale Genome-Wide Association Studies (GWAS). The primary method of Mendelian Randomization utilized in this study is the Inverse Variance Weighted (IVW) method. Results: Our findings indicate a bidirectional causal relationship between migraine and insomnia. In the discovery set, migraine had a significant effect on insomnia (OR=1.02, 95% CI=1.02 (1.01-1.03), PIVW=5.30E-04). However, this effect was not confirmed in the validation set (OR=1.03, 95% CI=1.03 (0.87-1.21), PIVW=0.77). Insomnia also had a significant effect on migraine (OR=1.02, 95% CI=1.02 (0.01-1.03), PIVW=2.67E-08), and this effect was validated in the validation set (OR=2.30, 95% CI=2.30 (1.60-3.30), PIVW=5.78E-06). Conclusion: This study provides meaningful insights into the bidirectional causality between migraine and insomnia, highlighting a complex interplay between these conditions. While our findings advance the understanding of the relationship between migraine and insomnia, they also open up new avenues for further research. The results underscore the need for considering both conditions in clinical and therapeutic strategies.

mTORC1 - TFEB pathway was involved in sodium arsenite induced lysosomal alteration, oxidative stress and genetic damage in BEAS-2B cells.

The mechanistic target of rapamycin (RAPA) complex 1 (mTORC1) - transcription factor EB (TFEB) pathway plays a crucial role in response to nutritional status, energy and environmental stress for maintaining cellular homeostasis. But there is few reports on its role in the toxic effects of arsenic exposure and the related mechanisms. Here, we show that the exposure of bronchial epithelial cells (BEAS-2B) to sodium arsenite promoted the activation of mTORC1 (p-mTORC1) and the inactivation of TFEB (p-TFEB), the number and activity of lysosomes decreased, the content of reduced glutathione (GSH) and superoxide dismutase (SOD) decreased, the content of malondialdehyde (MDA) increased, the DNA and chromosome damage elevated. Further, when mTORC1 was inhibited with RAPA, p-mTORC1 and p-TFEB down-regulated, GSH and SOD increased, MDA decreased, the DNA and chromosome damage reduced significantly, as compared with the control group. Our data revealed for the first time that mTORC1 - TFEB pathway was involved in sodium arsenite induced lysosomal alteration, oxidative stress and genetic damage in BEAS-2B cells, and it may be a potential intervention target for the toxic effects of arsenic.

New method for calculating the windward area of irregular fragments.

Average windward area is an important index for calculating the trajectory, velocity attenuation and terminal effect of explosive fragments. In order to solve the problems that existing theoretical method cannot calculate windward area of irregular fragment and experiment method is not convenient for automatic calculation and has low accuracy, a Monte Carlo subdivision projection simulation algorithm is proposed. The average windward area of arbitrary shaped fragments can be obtained with coordinate translation, random rotation, plane projection, convex-hull triangulation, concave boundary searching and sorting with maximum edge length constraint, subdivision area calculation, and averaging by thousands of cycles. Results show that projection area obtained by the subdivision projection algorithm is basically the same as that obtained by software method of computer aided design. Moreover, the maximum calculation error of the algorithm is less than 7%, and its accuracy is much higher than that of the equivalent ellipsoid method. The average windward area calculated by the Monte Carlo subdivision projection simulation algorithm is consistent with theoretical formula for prefabricated fragments, and the error is less than 3%. The convergence and accuracy of the Monte Carlo subdivision projection algorithm are better than those of the icosahedral uniform orientation method.

Comprehensive analysis of genetic associations and single-cell expression profiles reveals potential links between migraine and multiple diseases: a phenome-wide association study.

Migraine is a common neurological disorder that affects more than one billion people worldwide. Recent genome-wide association studies have identified 123 genetic loci associated with migraine risk. However, the biological mechanisms underlying migraine and its relationships with other complex diseases remain unclear. We performed a phenome-wide association study (PheWAS) using UK Biobank data to investigate associations between migraine and 416 phenotypes. Mendelian randomization was employed using the IVW method. For loci associated with multiple diseases, pleiotropy was tested using MR-Egger. Single-cell RNA sequencing data was analyzed to profile the expression of 73 migraine susceptibility genes across brain cell types. qPCR was used to validate the expression of selected genes in microglia. PheWAS identified 15 disorders significantly associated with migraine, with one association detecting potential pleiotropy. Single-cell analysis revealed elevated expression of seven susceptibility genes (including ZEB2, RUNX1, SLC24A3, ANKDD1B, etc.) in brain glial cells. And qPCR confirmed the upregulation of these genes in LPS-treated microglia. This multimodal analysis provides novel insights into the link between migraine and other diseases. The single-cell profiling suggests the involvement of specific brain cells and molecular pathways. Validation of gene expression in microglia supports their potential role in migraine pathology. Overall, this study uncovers pleiotropic relationships and the biological underpinnings of migraine susceptibility.

MCF-7 cell - derived exosomes were involved in protecting source cells from the damage caused by tributyltin chloride via transport function.

Tributyltin chloride (TBTC) is a ubiquitous environmental pollutant with various adverse effects on human health. Exosomes are cell - derived signaling and substance transport vesicles. This investigation aimed to explore whether exosomes could impact the toxic effects caused by TBTC via their transport function. Cytotoxicity, DNA and chromosome damage caused by TBTC on MCF-7 cells were analyzed with CCK-8, flow cytometry, comet assay and micronucleus tests, respectively. Exosomal characterization and quantitative analysis were performed with ultracentrifugation, transmission electron microscope (TEM) and bicinchoninic acid (BCA) methods. TBTC content in exosomes was detected with Liquid Chromatography-Mass Spectrometry (LC-MS). The impacts of exosomal secretion on the toxic effects of TBTC were analyzed. Our data indicated that TBTC caused significant cytotoxicity, DNA and chromosome damage effects on MCF-7 cells, and a significantly increased exosomal secretion. Importantly, TBTC could be transported out of MCF-7 cells by exosomes. Further, when exosomal secretion was blocked with GW4869, the toxic effects of TBTC were significantly exacerbated. We concluded that TBTC promoted exosomal secretion, which in turn transported TBTC out of the source cells to alleviate its toxic effects. This investigation provided a novel insight into the role and mechanism of exosomal release under TBTC stress.

Terminal dual-labeling of a transcribed RNA.

We report here a site-specific terminal dual-labeling strategy for a transcribed RNA. The combination of 5'-thiophosphoryl and 3'-amino functionalities enables efficient RNA dual labeling with different fluorophores at both 5'- and 3'-terminal positions specifically. This dual-labeling strategy is applied to pre-miRNA for construction of molecular beacons. The RNA beacons in their native hairpin formation bring the fluorophore and quencher groups into close proximity, leading to fluorescence quenching by FRET effect. Ribonuclease (dicer enzyme or micrococcal nuclease) can efficiently cleave RNA beacons leading to concentration- and time-dependent fluorescence increase. The dual-labeling strategy for transcribed RNAs involves only commercially available reagents, enzymes and native RNA, making it more accessible for general applications.

Effect of metal oxides on the light radiation intensity of Ba(NO3)2/Mg-containing pyrotechnic mixtures combustion.

Studying how to improve the performance of illuminating agents to meet the requirements of ammunition miniaturization of great importance. In this study, a simple method for increasing light radiation intensity through the adding of metal oxides was developed and tested. Results revealed that the metal oxides had a very strong effect on the light radiation intensity of the reaction system. Optical radiation intensity increased by 17.8%, - 5.4% and 25.9% after the addition 5% of MgO, Al2O3 and BaO to the Ba(NO3)2/Mg reaction system, respectively. This phenomenon may be related to the light radiation characteristics and reactivity of the metal oxide itself, as well as the temperature at which the added metal oxide can be excited to radiate light intensity.

In vitro expansion of DNA triplet repeats with bulge binders and different DNA polymerases.

The expansion of DNA repeat sequences is associated with many genetic diseases in humans. Simple bulge DNA structures have been implicated as intermediates in DNA slippage within the DNA repeat regions. To probe the possible role of bulged structures in DNA slippage, we designed and synthesized a pair of simple chiral spirocyclic compounds [Xi Z, Ouyang D & Mu HT (2006) Bioorg Med Chem Lett 16, 1180-1184], DDI-1A and DDI-1B, which mimic the molecular architecture of the enediyne antitumor antibiotic neocarzinostatin chromophore. Both compounds strongly stimulated slippage in various DNA repeats in vitro. Enhanced slippage synthesis was found to be synchronous for primer and template. CD spectra and UV thermal stability studies supported the idea that DDI-1A and DDI-1B exhibited selective binding to the DNA bulge and induced a significant conformational change in bulge DNA. The proposed mechanism for the observed in vitro expansion of long DNA is discussed.

A synergistic effect of Cu(2+) and norbixin on DNA damage.

Annatto and its derivatives are members of carotenoids with long-chain conjugated polyenes, which are widely used as food additives and antioxidant. However, carotenoids can also act as pro-oxidant under certain circumstances. To explore the biochemical behavior of annatto and its derivatives, the DNA damage effects by norbixin to the copper(II) ions mediated DNA damage was evaluated herein. It has been found that norbixin was capable of promoting copper(II) caused DNA damage on supercoiled plasmid DNA, whereas the long-chain saturated system such as lauric acid did not. These DNA damage showed strong dependency on both the concentration of norbixin and the interaction time. For the mechanism of damage promotion by norbixin, the long-chain conjugated polyenes unit may participate in the reductive reaction of copper(II) ion to generate free radical, and gave stronger DNA damage through the interactions between DNA and the radicalized carotenoids. The control experiments showed that the redox cycle between Cu(I) and Cu(II), hydroxyl radical, singlet oxygen and hydrogen peroxide may play essential roles in the cleavage reaction.

Estimation of Residual Stress in Selective Laser Melting of a Zr-Based Amorphous Alloy.

An accurate estimation of residual stresses is crucial to ensure dimensional accuracy and prevent premature fatigue failure of 3D printed components. Different from their crystalline counterparts, the effect of residual stress would be worse for amorphous alloys owing to their intrinsic brittleness with low fracture toughness. However, the generation of residual stress and its performance in 3D printed amorphous alloy components still remain unclear. Here, a finite element method combined with experiments and theoretical analyses was introduced to estimate the residual stress in selective laser melting of a Zr-based amorphous alloy. The results revealed that XY cross scanning strategy exhibits relatively low residual stress by comparison with X and Y strategies, and the residual stress becomes serious with increasing bar thickness. The residual stress, on the other hand, could be tuning by annealing or preheating the substrate. The above scenario is thoroughly understood according to the temperature gradient mechanism and its effect on microstructure evaluation.

Low temperature deformation behavior of an electromagnetically bulged 5052 aluminum alloy.

The fundamental understanding of the deformation behavior of electromagnetically formed metallic components under extreme conditions is important. Here, the effect of low temperature on the deformation behavior of an electromagnetically-bulged 5052 aluminum alloy was investigated through uniaxial tension. We found that the Portevin-Le Chatelier Effect, designated by the serrated characteristic in stress-strain curves, continuously decays until completely disappears with decreasing temperature. The physical origin of the phenomenon is rationalized on the basis of the theoretical analysis and the Monte Carlo simulation, which reveal an increasing resistance to dislocation motion imposed by lowering temperature. The dislocations are captured completely by solute atoms at -50 °C, which results in the extinction of Portevin-Le Chatelier. The detailed mechanism responsible for this process is further examined through Monte Carlo simulation.

Flexible DNA junction assisted efficient construction of stable gene nanoparticles for gene delivery.

A flexible DNA junction was designed to assist the construction of stable gene nanoparticles with multiple target gene copies, which can be used as efficient gene cargo with successful expression in eukaryotic cells for gene delivery.

Efficient construction of stable gene nanoparticles through polymerase chain reaction with flexible branched primers for gene delivery.

Flexible branched primers were designed to construct stable gene nanoparticles with multiple target gene copies through polymerase chain reaction, which can be used as an efficient transcription template in eukaryotic cells for gene delivery.

Self-assembled controllable virus-like nanorods as templates for construction of one-dimensional organic-inorganic nanocomposites.

Virus-like assemblies with controllable size and surface groups can be used as efficient templates for the controllable synthesis of CdS nanorods, which represents a new strategy for controllable preparation of one-dimensional (1D) organic-inorganic nanocomposites.

Interaction of bulged DNA with leucine-containing mimics of NCS-chrom.

Synthesis of chiral spirocyclic helical compounds containing leucine that mimic the molecular architecture of the potent DNA bulge binder obtained from the natural product metabolite NCSi-gb has been accomplished. The interaction between the compounds and DNA was studied by circular dichroism (CD) method. The results suggested that the two synthetic diastereoisomers specifically targeted the bulge site of DNA and induced conformational change of bulged DNA greatly.

Stimulation on DNA triplet repeat strand slippage synthesis by the designed spirocycles.

The designed simpler chiral spirocyclic helical compounds that mimic the molecular architecture of the DNA bulge binder NCSi-gb have been prepared. It has been found that the synthesized spirocyclic compounds have strong stimulation effect on DNA slippage synthesis. Their stimulation activities on DNA strand slippage suggest that they may bind to or induce the formation of a non Watson-Crick structure during in vitro replication of DNA triplet repeats.