An in-situ-tag-generation proximity labeling technology for recording cellular interactions.

Obtaining information about cellular interactions is fundamental to the elucidation of physiological and pathological processes. Proximity labeling technologies have been widely used to report cellular interactions in situ; however, the reliance on addition of tag molecules typically restricts their application to regions where tags can readily diffuse, while the application in, for example, solid tissues, is susceptible. Here, we propose an "in-situ-tag-generation mechanism" and develop the GalTag technology based on galactose oxidase (GAO) for recording cellular interactions within three-dimensional biological solid regions. GAO mounted on bait cells can in situ generate bio-orthogonal aldehyde tags as interaction reporters on prey cells. Using GalTag, we monitored the dynamics of cellular interactions and assessed the targeting ability of engineered cells. In particular, we recorded, for the first time, the footprints of Bacillus Calmette-Guérin (BCG) invasion into the bladder tissue of living mice, providing a valuable perspective to elucidate the anti-tumor mechanism of BCG.

3D Multimodal Fusion Network with Disease-induced Joint Learning for Early Alzheimer's Disease Diagnosis.

Multimodal neuroimaging provides complementary information critical for accurate early diagnosis of Alzheimer's disease (AD). However, the inherent variability between multimodal neuroimages hinders the effective fusion of multimodal features. Moreover, achieving reliable and interpretable diagnoses in the field of multimodal fusion remains challenging. To address them, we propose a novel multimodal diagnosis network based on multi-fusion and disease-induced learning (MDL-Net) to enhance early AD diagnosis by efficiently fusing multimodal data. Specifically, MDL-Net proposes a multi-fusion joint learning (MJL) module, which effectively fuses multimodal features and enhances the feature representation from global, local, and latent learning perspectives. MJL consists of three modules, global-aware learning (GAL), local-aware learning (LAL), and outer latent-space learning (LSL) modules. GAL via a self-adaptive Transformer (SAT) learns the global relationships among the modalities. LAL constructs local-aware convolution to learn the local associations. LSL module introduces latent information through outer product operation to further enhance feature representation. MDL-Net integrates the disease-induced region-aware learning (DRL) module via gradient weight to enhance interpretability, which iteratively learns weight matrices to identify AD-related brain regions. We conduct the extensive experiments on public datasets and the results confirm the superiority of our proposed method. Our code will be available at: https://github.com/qzf0320/MDL-Net.

Selenium-based metabolic oligosaccharide engineering strategy for quantitative glycan detection.

Metabolic oligosaccharide engineering (MOE) is a classical chemical approach to perturb, profile and perceive glycans in physiological systems, but probes upon bioorthogonal reaction require accessibility and the background signal readout makes it challenging to achieve glycan quantification. Here we develop SeMOE, a selenium-based metabolic oligosaccharide engineering strategy that concisely combines elemental analysis and MOE,enabling the mass spectrometric imaging of glycome. We also demonstrate that the new-to-nature SeMOE probes allow for detection, quantitative measurement and visualization of glycans in diverse biological contexts. We also show that chemical reporters on conventional MOE can be integrated into a bifunctional SeMOE probe to provide multimodality signal readouts. SeMOE thus provides a convenient and simplified method to explore the glyco-world.

Decreased HAT1 expression in granulosa cells disturbs oocyte meiosis during mouse ovarian aging.

BACKGROUND: With advanced maternal age, abnormalities during oocyte meiosis increase significantly. Aneuploidy is an important reason for the reduction in the quality of aged oocytes. However, the molecular mechanism of aneuploidy in aged oocytes is far from understood. Histone acetyltransferase 1 (HAT1) has been reported to be essential for mammalian development and genome stability, and involved in multiple organ aging. Whether HAT1 is involved in ovarian aging and the detailed mechanisms remain to be elucidated. METHODS: The level of HAT1 in aged mice ovaries was detected by immunohistochemical and immunoblotting. To explore the function of HAT1 in the process of mouse oocyte maturation, we used Anacardic Acid (AA) and small interfering RNAs (siRNA) to culture cumulus-oocyte complexes (COCs) from ICR female mice in vitro and gathered statistics of germinal vesicle breakdown (GVBD), the first polar body extrusion (PBE), meiotic defects, aneuploidy, 2-cell embryos formation, and blastocyst formation rate. Moreover, the human granulosa cell (GC)-like line KGN cells were used to investigate the mechanisms of HAT1 in this progress. RESULTS: HAT1 was highly expressed in ovarian granulosa cells (GCs) from young mice and the expression of HAT1 was significantly decreased in aged GCs. AA and siRNAs mediated inhibition of HAT1 in GCs decreased the PBE rate, and increased meiotic defects and aneuploidy in oocytes. Further studies showed that HAT1 could acetylate Forkhead box transcription factor O1 (FoxO1), leading to the translocation of FoxO1 into the nucleus. Resultantly, the translocation of acetylated FoxO1 increased the expression of amphiregulin (AREG) in GCs, which plays a significant role in oocyte meiosis. CONCLUSION: The present study suggests that decreased expression of HAT1 in GCs is a potential reason corresponding to oocyte age-related meiotic defects and provides a potential therapeutic target for clinical intervention to reduce aneuploid oocytes.

Effect of the Functional VP1 Unique Region of Human Parvovirus B19 in Causing Skin Fibrosis of Systemic Sclerosis.

Human parvovirus B19 (B19V) is a single-stranded non-enveloped DNA virus of the family Parvoviridae that has been associated with various autoimmune disorders. Systemic sclerosis (SSc) is an autoimmune connective tissue disorder with high mortality and has been linked to B19V infection. However, the precise mechanism underlying the B19V contribution to the development of SSc remains uncertain. This study investigated the impacts of the functional B19V-VP1 unique region (VP1u) in macrophages and bleomycin (BLE)-induced SSc mice. Cell experimental data showed that significantly decreased viability and migration of both B19V-VP1u-treated U937 and THP-1 macrophages are detected in the presence of celastrol. Significantly increased MMP9 activity and elevated NF-kB, MMP9, IL-6, TNF-α, and IL-1ß expressions were detected in both B19V-VP1u-treated U937 and THP-1 macrophages. Conversely, celastrol revealed an inhibitory effect on these molecules. Notably, celastrol intervened in this pathogenic process by suppressing the sPLA2 activity of B19V-VP1u and subsequently reducing the inflammatory response. Notably, the administration of B19V-VP1u exacerbated BLE-induced skin fibrosis in mice, with augmented expressions of TGF-ß, IL-6, IL-17A, IL-18, and TNF-α, ultimately leading to α-SMA and collagen I deposits in the dermal regions of BLE-induced SSc mice. Altogether, this study sheds light on parvovirus B19 VP1u linked to scleroderma and aggravated dermal fibrosis.

Decreased LONP1 expression contributes to DNA damage and meiotic defects in oocytes.

Meiotic defects in oocytes are the primary reason for decreased female fertility with advanced maternal age. In this study, we revealed that decreased expression of ATP-dependent Lon peptidase 1 (LONP1) in aged oocytes and oocyte-specific depletion of LONP1 disrupt oocyte meiotic progression accompanying with mitochondrial dysfunction. In addition, LONP1 downregulation increased oocyte DNA damage. Moreover, we demonstrated that splicing factor proline and glutamine rich directly interacts with LONP1 and mediate the effect of LONP1 depletion on meiotic progression in oocytes. In summary, our data suggest that decreased expression of LONP1 is involved in advanced maternal age-related meiosis defects and that LONP1 represents a new therapeutic target to improve aged oocyte quality.

Granulosa cell mevalonate pathway abnormalities contribute to oocyte meiotic defects and aneuploidy.

With aging, abnormalities during oocyte meiosis become more prevalent. However, the mechanisms of aging-related oocyte aneuploidy are not fully understood. Here we performed Hi-C and SMART-seq of oocytes from young and old mice and reveal decreases in chromosome condensation and disrupted meiosis-associated gene expression in metaphase I oocytes from aged mice. Further transcriptomic analysis showed that meiotic maturation in young oocytes was correlated with robust increases in mevalonate (MVA) pathway gene expression in oocyte-surrounding granulosa cells (GCs), which was largely downregulated in aged GCs. Inhibition of MVA metabolism in GCs by statins resulted in marked meiotic defects and aneuploidy in young cumulus-oocyte complexes. Correspondingly, supplementation with the MVA isoprenoid geranylgeraniol ameliorated oocyte meiotic defects and aneuploidy in aged mice. Mechanically, we showed that geranylgeraniol activated LHR/EGF signaling in aged GCs and enhanced the meiosis-associated gene expression in oocytes. Collectively, we demonstrate that the MVA pathway in GCs is a critical regulator of meiotic maturation and euploidy in oocytes, and age-associated MVA pathway abnormalities contribute to oocyte meiotic defects and aneuploidy.

Effects of ultrasonic vibration on microstructure and mechanical properties of 1Cr12Ni3MoVN alloy fabricated by directed energy deposition.

Due to the rapid melting and solidification during directed energy deposition (DED) process, the defects and columnar crystals are likely to generate in the deposition layers, which reduce the quality and performance of the whole parts. Therefore, in order to improve the microstructure and mechanical properties of 1Cr12Ni3MoVN alloy manufactured by DED method, ultrasonic vibration (UV) has been employed to assist directed energy deposition process in this work. The results indicate that the high-intensity ultrasonic vibration can weaken the epitaxy growth tendency of crystal grains, and significantly improve plasticity while keeping an approximate strength. In addition, a two-dimensional numerical model is established to simulate the effect of ultrasonic vibration in the molten pool. The simulation results show that ultrasonic vibration remarkably improves the flow velocity and pressure in the molten pool, inducing the cavitation effect that breaks dendritic crystal and affects crystal characteristics. Meanwhile, the acoustic streaming effect changes the thermodynamic conditions and promotes high-temperature diffusion, which uniforms temperature distribution and reduces the temperature gradient in the molten pool. Thus the reduced temperature gradient G and raised solidification growth rate R promote the formation of fine equiaxed crystal characteristics after UV treatment. The product G × R increases and the ratio G/R decreases after UV treatment, resulting in the formation of fine equiaxed crystals.

How does public environmental supervision affect the industrial structure optimization?

Based on the panel data of 278 prefecture-level cities in China from 2005 to 2017, this study takes the environmental information disclosure policy implemented in 2008 as a quasi-natural experiment and uses the difference-in-difference method to examine the impact of public environmental supervision on industrial structure upgrading. This study found that public environmental supervision has indeed significantly promoted the industrial structure optimization. It is noteworthy that, the impact of public environmental supervision on industrial structure optimization is heterogeneous across regions and corresponding to the intensity of environmental regulations. Public environmental supervision in the eastern region has significantly promoted the optimization of the industrial structure, while the promotion effect in the central and western regions is not obvious. Besides, the effect of public environmental supervision is more significant in the cities with relatively tight traditional environmental regulations. The mechanism analysis shows that public environmental supervision promotes industrial structure upgrading mainly by influencing the level of urban technological innovation, and the level of technological innovation plays an intermediary role between public environmental supervision and industrial structure optimization. This study has important reference significance for further improving the environmental information disclosure system and constructing an environmental governance system with the government as the leading body, enterprises as the main body, and the public as the common participation.

Maternal circadian disruption before pregnancy impairs the ovarian function of female offspring in mice.

Circadian disturbance brought on by shift employment, nighttime light pollution, and other factors is quite prevalent in contemporary culture. However, the effect of maternal circadian disruption before pregnancy on the reproduction of offspring in mice requires further research. Herein, we exposed female ICR mice to constant light to establish a model of preconceptional circadian disruption and then checked the ovarian function of female offspring (named the CLE group below). Our results revealed obesity, abnormal lipid metabolism and earlier puberty onset in the CLE group. Additionally, impaired ovarian follicle development, oocyte quality and preimplantation embryo development were shown in the CLE group. Moreover, the expression levels of Gnrh1 in the hypothalamus and Cyp17a1, Bmper, Bdnf and Lyve1 in ovaries, as well as circadian clock genes, including Clock, Cry1, Nr1d2 and Per2, were significantly downregulated in the CLE group. Mechanistically, immune responses, including the interleukin-17 (IL-17) signalling pathway, cytokine-cytokine receptor interaction and the chemokine signalling pathway, were altered in the CLE group, which may be responsible for the damaged ovarian function.

Deep Mobile Linguistic Therapy for Patients with ASD.

Autistic spectrum disorder (ASD) is one of the most complex groups of neurobehavioral and developmental conditions. The reason is the presence of three different impaired domains, such as social interaction, communication, and restricted repetitive behaviors. Some children with ASD may not be able to communicate using language or speech. Many experts propose that continued therapy in the form of software training in this area might help to bring improvement. In this work, we propose a design of software speech therapy system for ASD. We combined different devices, technologies, and features with techniques of home rehabilitation. We used TensorFlow for Image Classification, ArKit for Text-to-Speech, Cloud Database, Binary Search, Natural Language Processing, Dataset of Sentences, and Dataset of Images with two different Operating Systems designed for Smart Mobile devices in daily life. This software is a combination of different Deep Learning Technologies and makes Human-Computer Interaction Therapy very easy to conduct. In addition, we explain the way these were connected and put to work together. Additionally, we explain in detail the architecture of software and how each component works together as an integrated Therapy System. Finally, it allows the patient with ASD to perform the therapy anytime and everywhere, as well as transmitting information to a medical specialist.

Deep Neural Network Framework Based on Word Embedding for Protein Glutarylation Sites Prediction.

In recent years, much research has found that dysregulation of glutarylation is associated with many human diseases, such as diabetes, cancer, and glutaric aciduria type I. Therefore, glutarylation identification and characterization are essential tasks for determining modification-specific proteomics. This study aims to propose a novel deep neural network framework based on word embedding techniques for glutarylation sites prediction. Multiple deep neural network models are implemented to evaluate the performance of glutarylation sites prediction. Furthermore, an extensive experimental comparison of word embedding techniques is conducted to utilize the most efficient method for improving protein sequence data representation. The results suggest that the proposed deep neural networks not only improve protein sequence representation but also work effectively in glutarylation sites prediction by obtaining a higher accuracy and confidence rate compared to the previous work. Moreover, embedding techniques were proven to be more productive than the pre-trained word embedding techniques for glutarylation sequence representation. Our proposed method has significantly outperformed all traditional performance metrics compared to the advanced integrated vector support, with accuracy, specificity, sensitivity, and correlation coefficient of 0.79, 0.89, 0.59, and 0.51, respectively. It shows the potential to detect new glutarylation sites and uncover the relationships between glutarylation and well-known lysine modification.

Human Endometrium-Derived Adventitial Cell Spheroid-Loaded Antimicrobial Microneedles for Uterine Regeneration.

Asherman's syndrome (AS) occurs as a consequence of severe damage to the endometrial basalis, usually leading to menstrual abnormalities, infertility, and recurrent miscarriage in women. Currently, human endometrium-derived adventitial cells (En-ADVs) are considered ideal seed cells with high pluripotency for regenerative medicine. However, critical issues such as noninvasive repair of tissues, targeting of native stem cells, and continuous action in the injured sites are not well resolved. Herein, En-ADV spheroid-loaded hierarchical microneedles (MN/En-ADV) for in situ intrauterine repair are developed. The flexible microneedles are fabricated with gelatin methacryloyl and lactoferrin, imparting the characteristics of rapid degradation and antimicrobial activity. Benefiting from an array of microwells on microneedles, En-ADVs can rapidly form 3D cell spheroids, which display higher potential for cell proliferation, differentiation, and migration than dissociated cells. With the application of MN/En-ADV, the repaired uteri show well-defined myometrial regeneration, angiogenesis, and an increase of endometrial receptivity in a rat AS model. Notably, embryos are able to implant in the reconstructed sites and remain viable, indicating that this system promotes the restoration of both normal morphology and reproductive function in the injured uterus. It is anticipated that multifunctional MN/En-ADV can be an ideal candidate for versatile in situ tissue regeneration.

The mitochondrial protease LONP1 maintains oocyte development and survival by suppressing nuclear translocation of AIFM1 in mammals.

BACKGROUND: Oogenesis is a fundamental process of human reproduction, and mitochondria play crucial roles in oocyte competence. Mitochondrial ATP-dependent Lon protease 1 (LONP1) functions as a critical protein in maintaining mitochondrial and cellular homeostasis in somatic cells. However, the essential role of LONP1 in maintaining mammalian oogenesis is far from elucidated. METHODS: Using conditional oocyte Lonp1-knockout mice, RNA sequencing (RNA-seq) and coimmunoprecipitation/liquid chromatography-mass spectrometry (Co-IP/LC-MS) technology, we analysed the functions of LONP1 in mammalian oogenesis. FINDINGS: Conditional knockout of Lonp1 in mouse oocytes in both the primordial and growing follicle stages impairs follicular development and causes progressive oocyte death, ovarian reserve loss, and infertility. LONP1 directly interacts with apoptosis inducing factor mitochondria-associated 1 (AIFM1), and LONP1 ablation leads to the translocation of AIFM1 from the cytoplasm to the nucleus, causing apoptosis in mouse oocytes. In addition, women with pathogenic variants of LONP1 lack large antral follicles (>10 mm) in the ovaries, are infertile and present premature ovarian insufficiency. INTERPRETATION: We demonstrated the function of LONP1 in regulating oocyte development and survival, and in-depth analysis of LONP1 will be crucial for elucidating the mechanisms underlying premature ovarian insufficiency. FUNDING: This work was supported by grants from the National Key Research and Development Program of China (2018YFC1004701), the National Nature Science Foundation of China (82001629, 81871128, 81571391, 81401166, 82030040), the Jiangsu Province Social Development Project (BE2018602), the Jiangsu Provincial Medical Youth Talent (QNRC2016006), the Youth Program of the Natural Science Foundation of Jiangsu Province (BK20200116) and Jiangsu Province Postdoctoral Research Funding (2021K277B).

Growth hormone ameliorates the age-associated depletion of ovarian reserve and decline of oocyte quality via inhibiting the activation of Fos and Jun signaling.

Oocyte quality typically begins to decline with aging, which contributes to subfertility and infertility. However, there is still no effective treatment to restore the ovarian reserve and improve aged-oocyte quality. According to the present study, growth hormone (GH) secretion changes with maternal age in female mice. After intraperitoneal injection with GH (1 mg/kg body weight) every two days for two months, the 10-month-old mice showed a better ovarian reserve and oocyte quality than control mice. GH treatment decreased the occurrence rate of aneuploidy caused by spindle/chromosome defects. Additionally, the single oocyte transcriptome analysis indicated that GH decreased the expression of apoptosis-related genes in oocytes. It was also observed that GH treatment reduced the expression of γH2AX and apoptosis of aged oocytes via decreasing the activation of Fos and Jun. Collectively, our results indicate that GH treatment is an effective way to reverse the age-associated depletion of ovarian reserve and the decline of oocyte quality by decreasing apoptosis.

[miR-21 promotes expression of NF-κB and NLRP3 in mouse RAW264.7 cells by inhibiting A20].

Objective To investigate the effect of miR-21 on the expression of NF-κB and NLR family pyrin domain containing 3 (NLRP3) in RAW264.7 cells stimulated by lipopolysaccharide (LPS) and its possible mechanism. Methods Real-time fluorescent quantitative PCR was used to detect the expression of miR-21 after RAW264.7 cells were stimulated with 100 ng/mL LPS for 24 hours. miR-21 inhibitors or mimics were transfected to regulate the expression of miR-21 in RAW264.7 cells. After miR-21 inhibitors or mimics were stimulated by LPS, real-time quantitative PCR was used to detect its effect on the mRNA expression of interleukin 6 (IL-6), tumor necrosis factor α (TNF-α) and IL-1ß, and Western blotting was used to detect the effects on the expression of NF-κB, NLRP3 and A20 proteins. Results The expression of miR-21 significantly increased when RAW264.7 cells were stimulated by LPS. The expression of IL-6, TNF-α, IL-1ß, NF-κB and NLRP3 was raised when the expression of miR-21 was up-regulated. The expression of IL-6, TNF-α, IL-1ß, NF-κB and NLRP3 was reduced when the expression of miR-21 was down-regulated. miR-21 targeted the inhibition of A20 expression. Conclusion miR-21 can promote the expression of NF-κB and NLRP3 in RAW264.7 cells and its mechanism may be related to the targeted inhibition of A20.

[Advances in the study of ovarian dysfunction with aging].

Societal changes regarding the role of women have significant impacts on women's willingness and the timing of childbearing. Ovarian reserve in woman typically begins to decline at the age of 35, and it is primarily characterized by a reduction in the number of ovarian follicles and a decline in oocyte quality. The clinical diagnosis of ovarian insufficiency relies on multiple variables including changes of follicle stimulating hormone (FSH), serum anti-Müllerian hormone (AMH), inhibin B, antral follicle count, menstruation and age. It is proven that ovarian cells demonstrate dysfunction associated with aging including mitochondrial dysfunction, telomere shortening, impaired DNA repair, epigenetic changes and metabolic/energetic disorders. In this review, we introduce the clinical diagnosis and management of ovarian insufficiency. We mainly discuss the molecular mechanism and potential interventions. We are optimistic that this information and knowledge will inform the important decisions for women and society regarding childbearing.

A novel mechanism of pre-transplant insulin resistance contributing to post-transplant complications: Cyclosporin A-induced O-GlcNAcylation.

Pre-transplant insulin resistance has been proved to be an important risk factor for organ transplantation, predicting increased post-transplant complications and worse survival outcomes. However, the underlying mechanism is still unclear. Cyclosporin A (CsA) is widely used as an immunosuppressant after organ transplantation, while emerging evidence has shown that CsA increases the risk of post-transplant complications. Thus, in this study, using a cellular model of palmitate-induced insulin resistance, we evaluate the effect of CsA on apoptosis in skeletal muscle C2C12 cells with palmitate-induced insulin resistance. Western blot and flow cytometric analysis showed that CsA induced apoptosis in insulin-resistant C2C12 cells. Mechanistically, a sustained increase of global protein O-GlcNAcylation was observed after CsA treatment, and suppression of protein O-GlcNAcylation with its inhibitors (alloxan or 5-oxo-6-diazo-norleucine) resulted in decreased O-GlcNAcylation levels and apoptosis. Furthermore, CsA increased mitochondrial membrane potential and intracellular ROS production in insulin-resistant C2C12 cells, and inhibition of ROS production with SS-31 suppressed CsA-induced O-GlcNAcylation. In summary, our results suggest that CsA treatment induced apoptosis in insulin-resistant C2C12 cells, partly via CsA-induced ROS production and resultant O-GlcNAcylation, indicating that O-GlcNAcylation serves as a potent therapeutical target for organ transplantation.

Comparison of the glycopattern alterations of mitochondrial proteins in cerebral cortex between rat Alzheimer's disease and the cerebral ischemia model.

Alzheimer's disease (AD) and ischemic brain injury are two major neurodegenerative diseases. Mitochondrial dysfunction commonly occurs in AD and ischemic brain injury. Currently, little attention has been paid to the glycans on mitochondrial glycoproteins, which may play vital roles during the process of mitochondrial dysfunction. The aim of this study was to illustrate and compare the glycopattern alterations of mitochondrial glycoproteins extracted from the cerebral cortex of the rat models of these two diseases using High-throughput lectin microarrays. The results shown that the number of lectins with significant differences compared to normal brains was nine for the rat sporadic Alzheimer's disease (SAD) model and eighteen for the rat middle cerebral artery occlusion (MCAO) model. Interestingly, five lectins showed opposite expression patterns between the SAD and MCAO rat models. We conclude that glycopattern alterations of mitochondrial glycoproteins in the cerebral cortex may provide vital information to help understand mitochondrial dysfunction in AD and ischemic brain injury. In addition, glycans recognized by diverse lectins with opposite expression patterns between these two diseases hints at the different pathomechanisms of mitochondrial dysfunction in AD and ischemic brain injury.