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The viscosity within cells is a crucial microenvironmental factor, and sulfur dioxide (SO2 ) has essential functions in regulating cellular apoptosis and inflammation. Some evidence has been confirmed that changes in viscosity and overexposure of SO2 within the cell may cause detrimental effects including, but not limited to, respiratory and cardiovascular illnesses, inflammation, fatty liver, and various types of cancer. Therefore, precise monitoring of SO2 and viscosity in biological entities holds immense practical importance. Therefore, in this research, we developed a versatile fluorescent TCF-Cou that enables the dual detection of SO2 and viscosity in the living system. Probe TCF-Cou possessed a response to viscosity and SO2 through red and green emissions. The alteration of SO2 and viscosity levels in live cells and zebrafish were also monitored using probe TCF-Cou. We hope that this fluorescent probe could be a potential tool for revealing the related pathological and physiological processes through monitoring the changes in SO2 and viscosity.
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Colorantes Fluorescentes , Pez Cebra , Humanos , Animales , Células HeLa , Viscosidad , Dióxido de AzufreRESUMEN
Since their inception, rhodamine dyes have been extensively applied in biotechnology as fluorescent markers or for the detection of biomolecules owing to their good optical physical properties. Accordingly, they have emerged as a powerful tool for the visualization of living systems. In addition to fluorescence bioimaging, the molecular design of rhodamine derivatives with disease therapeutic functions (e.g., cancer and bacterial infection) has recently attracted increased research attention, which is significantly important for the construction of molecular libraries for diagnostic and therapeutic integration. However, reviews focusing on integrated design strategies for rhodamine dye-based diagnosis and treatment and their wide application in disease treatment are extremely rare. In this review, first, a brief history of the development of rhodamine fluorescent dyes, the transformation of rhodamine fluorescent dyes from bioimaging to disease therapy, and the concept of optics-based diagnosis and treatment integration and its significance to human development are presented. Next, a systematic review of several excellent rhodamine-based derivatives for bioimaging, as well as for disease diagnosis and treatment, is presented. Finally, the challenges in practical integration of rhodamine-based diagnostic and treatment dyes and the future outlook of clinical translation are also discussed.
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The concept of molecular design, integrating diagnostic and therapeutic functions, aligns with the general trend of modern medical advancement. Herein, we rationally designed the smart molecule ER-ZS for endoplasmic reticulum (ER)-targeted diagnosis and treatment in cell and animal models by combining hemicyanine dyes with ER-targeted functional groups (p-toluenesulfonamide). Owing to its ability to target the ER with a highly specific response to viscosity, ER-ZS demonstrated substantial fluorescence turn-on only after binding to the ER, independent of other physiological environments. In addition, ER-ZS, being a small molecule, allows for the diagnosis of nonalcoholic fatty liver disease (NAFLD) via liver imaging based on high ER stress. Importantly, ER-ZS is a typeâ I photosensitizer, producing O2 â - and â OH under light irradiation. Thus, after irradiating for a certain period, the photodynamic therapy inflicted severe oxidative damage to the ER of tumor cells in hypoxic (2 % O2 ) conditions and activated the unique pyroptosis pathway, demonstrating excellent antitumor capacity in xenograft tumor models. Hence, the proposed strategy will likely shed new light on integrating molecular optics for NAFLD diagnosis and cancer therapy.
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Carbocianinas , Neoplasias , Enfermedad del Hígado Graso no Alcohólico , Fotoquimioterapia , Animales , Humanos , Enfermedad del Hígado Graso no Alcohólico/diagnóstico por imagen , Enfermedad del Hígado Graso no Alcohólico/tratamiento farmacológico , Piroptosis , Colorantes/metabolismo , Viscosidad , Hígado/metabolismo , Retículo Endoplásmico/metabolismo , Estrés del Retículo Endoplásmico , Neoplasias/patologíaRESUMEN
Proteins have been perceived as being an intriguing modality of therapeutics for the treatment of intractable diseases in view of their superlative precision and versatility. Nonetheless, proteins' intrinsic characters, particularly their being hydrophilic macromolecules with unmethodical charges, have imposed the exceeding challenge of seeking transcellular trafficking into cells' interiors. To circumvent this drawback, we have attempted to employ triple-functional amine-reactive 4-(2-((2-(((4-nitrophenoxy)carbonyl)oxy)ethyl)disulfaneyl)ethoxy)-4-oxobutanoic acid for the efficient incorporation of the anionic carboxyl moiety into amine-enriched enzymes, resulting in overall negatively charged pro-enzymes. The resulting pro-enzymes could be readily electrostatically assembled with cationic species [for instance: block copolymers of poly(ethylene glycol)-polylysine] into core-shell architectural delivery nanoparticles for their facilitated endocytosis into cells. Noteworthy is the aforementioned carboxylation chemistry designed to allow facile reversal of the pro-enzymes to the original amine groups due to the thiolysis of intermediate disulfide linkage for subsequent cascade reactions in response to the cytosol-enriched glutathione. Therefore, cytosol-selective structural disassembly for the liberation and activation of the pro-enzymes was accomplished. Our subsequent investigations utilizing ribonuclease A and catalase as the model enzymes demonstrated appreciable transcellular transportation of the active enzymes to the cell interiors, exerting overwhelming cytotoxic potencies and H2O2 scavenging capacities, respectively. Hence, we reported an unprecedented redox-stimulated charge reversal strategy in engineering cytosol-activatable pro-enzymes, manifesting a simple and efficient approach in the manufacture of transcellular proteinic therapeutics, which should be highlighted to promote their wide availability for use with diverse functional proteins as molecular biological tools and precision therapeutics.
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Peróxido de Hidrógeno , Nanopartículas , Aminas , Glutatión , Nanopartículas/química , Polietilenglicoles/química , Polilisina/química , ProteínasRESUMEN
In light of immune facilities trafficking toward the pathological sites along upward gradient of immunostimulatory cytokines, a localized resiquimod (Toll-like receptor 7/8 agonist) release depot was manufactured for pursuit of precision immunostimulation toward intractable triple-negative breast carcinoma. In principle, resiquimod/poly(lactic-co-glycolic acid) microspheres were fabricated and embedded into injectable and biodegradable poly(ethylene glycol) (PEG)-based hydrogel. The subsequent investigations approved persistent retention of immunostimulatory resiquimod in tumors upon peritumoral administration, which consequently led to localized and consistent secretion of immunostimulatory cytokines. Initially, not only innate tumor phagocytosis but also adaptive antitumor immunities were successfully cultivated for in situ suppression of the growth of primary solid tumors, more importantly, capable of inhibiting distant pulmonary metastasis, as evidenced by observation of enormous lymphocytes selectively gathering in the pulmonary artery. Hence, our presented study provided an important clinical indication of using immunostimulatory drugs to activate potent innate and adaptive antitumor immunities for precision antitumor therapy. Further immunomodulatory strategies, such as checkpoint blockage and tumor immunogenicity, could also be complementary for development of advanced antitumor immunotherapeutics in treatment of a number of intractable tumors.
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Adyuvantes Inmunológicos/metabolismo , Imidazoles/metabolismo , Neoplasias de la Mama Triple Negativas/patología , Adyuvantes Inmunológicos/farmacología , Línea Celular Tumoral , Preparaciones de Acción Retardada , Humanos , Imidazoles/farmacología , Metástasis de la Neoplasia , Medicina de Precisión , Neoplasias de la Mama Triple Negativas/inmunologíaRESUMEN
MAIN CONCLUSION: Expression of GmPSKγ1 , a novel PSK-encoding gene from soybean, increases seed size and yield in transgenic plants by promoting cell expansion. Phytosulfokine-α (PSK-α), a sulfated pentapeptide hormone with the sequence YIYTQ, plays important roles in many aspects of plant growth and development. In this study, we identified a pair of putative precursor genes in soybean, GmPSKγ1 and -2, encoding a PSK-like peptide: PSK-γ. Similar to PSK-α in amino acid composition, the sequence of PSK-γ is YVYTQ, and the tyrosines undergo sulfonylation. Treatment of Arabidopsis seedlings with synthetic sulfated PSK-γ significantly enhanced root elongation, indicating that PSK-γ might be a functional analog of PSK-α. Expression pattern analysis revealed that the two GmPSKγ genes, especially GmPSKγ1, are primarily expressed in developing soybean seeds. Heterologous expression of GmPSKγ1 under the control of a seed-specific promoter markedly increased seed size and weight in Arabidopsis, and this promoting effect of PSK-γ on seed growth was further confirmed in transgenic tobacco constitutively expressing GmPSKγ1. Cytological analysis of transgenic Arabidopsis seeds revealed that PSK-γ promotes seed growth by inducing embryo cell expansion. In addition, expression analysis of downstream candidate genes suggested that PSK-γ signaling might regulate cell wall loosening to promote cell expansion in Arabidopsis seeds. Overall, our results shed light on the mechanism by which PSK-γ promotes seed growth, paving the way for the use of this new peptide for biotechnological improvement of crop seed/grain size and yield.
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Genes de Plantas/genética , Glycine max/genética , Proteínas de Plantas/genética , Semillas/crecimiento & desarrollo , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Clonación Molecular , Genes de Plantas/fisiología , Hormonas Peptídicas/genética , Filogenia , Proteínas de Plantas/fisiología , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Plantas Modificadas Genéticamente , Reacción en Cadena en Tiempo Real de la Polimerasa , Alineación de SecuenciaRESUMEN
A novel naphthalimide-based colorimetric and fluorescent turn-on chemosensor for Al3+ was synthesized and characterized with spectroscopic techniques. In MeOH solution, BPAM showed high selectivity and sensitivity to Al3+ by a 60-fold fluorescence enhancement and blue-shift absorption with visible color changes attributed to the contribution of chelation enhanced fluorescence (CHEF) and inhibition of intramolecular charge transfer (ICT). A 1:1 BPAM-Al3+ complex confirmed by job's plot and HRMS with a binding constant of 6.37 × 104 M- 1, and the detection limit for Al3+ was as low as 1.59 × 10- 7 M. BPAM was successfully applied in real sample detection and assessing the existence of Al3+ by a colorimetric method on filter paper. Furthermore, the fluorescent signals of BPAM were designed to construct an INHIBIT molecular logic gate.
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HSP90 plays important roles in multiple cellular stress responses. Here, two cytoplasmic HSP90 isoforms, ScHSP90α and ScHSP90ß, were identified from Siniperca chuatsi. Their cDNA and gDNA structures, amino acid sequence features, and sequence identities and phylogenetic analysis with other species were described. Their expression profiles during embryonic development in different tissues and under stressful conditions were analyzed using real-time quantitative PCR. During embryogenesis, transcripts of both genes were detected at low levels during the early developmental stages and were up-regulated from appearance of myomere for ScHSP90a and closure of blastopore for ScHSP90ß. ScHSP90α showed a tissue-specific variation with high expression in ovary and brain under non-stressed conditions, while ScHSP90ß was ubiquitously highly expressed in different tissues. Acute heat shock resulted in a strong up-regulation of ScHSP90α in heart, liver, and head kidney, while it only weakly induced ScHSP90ß in these tissues. ScHSP90α was also markedly induced in liver in a time-dependent manner under hypoxia, while the expression of ScHSP90ß was not affected by hypoxia. Additionally, Aeromonas hydrophila infection markedly augmented ScHSP90α in head kidney and spleen and mildly up-regulated ScHSP90ß in spleen, while suppressing ScHSP90ß in head kidney. These results suggest that ScHSP90α and ScHSP90ß are differently involved in embryogenesis and under different environmental conditions including high temperature, hypoxia, and bacterial infection. This study will benefit to further clarify the roles of fish HSP90 isoforms in embryogenesis and under stressful conditions and contribute to further study on enhancing stress tolerance and disease resistance of mandarin fish.
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Proteínas de Peces/genética , Proteínas HSP90 de Choque Térmico/genética , Perciformes/genética , Aeromonas hydrophila , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Desarrollo Embrionario/genética , Femenino , Enfermedades de los Peces/genética , Infecciones por Bacterias Gramnegativas/genética , Calor , Hipoxia/genética , Isoformas de Proteínas/genética , Estrés Fisiológico/genética , TranscriptomaRESUMEN
BACKGROUND: Obstructive sleep apnea (OSA) and its main feature, chronic intermit-tent hypoxia (IH) during sleep, is closely associated with insulin resistance (IR) and diabetes. Rimonabant can regulate glucose metabolism and improve IR. The present study aimed to assess the effect of IH and rimonabant on glucose metabolism and insulin sensitivity, and to explore the possible mechanisms. MATERIAL AND METHODS: Thirty-two rats were randomly assigned into 4 groups: Control group, subjected to intermittent air only; IH group, subjected to IH only; IH+NS group, subjected to IH and treated with normal saline; and IH+Rim group, subjected to IH and treated with 10 mg/kg/day of rimonabant. All rats were killed after 28 days of exposure. Then, the blood and skeletal muscle were collected. We measured fasting blood glucose levels, fasting blood insulin levels, and the expression of glucose transporter 4 (GLUT4) in both mRNA and protein levels in skeletal muscle. RESULTS: IH can slow weight gain, increase serum insulin level, and reduce insulin sensitivity in rats. The expressions of GLUT4 mRNA, total GLUT4, and plasma membrane protein of GLUT4 (PM GLUT4) in skeletal muscle were decreased. Rimonabant treatment was demonstrated to improve weight gain and insulin sensitivity of the rats induced by IH. Rimonabant significantly upregulated the expression of GLUT4 mRNA, PM GLUT4, and total GLUT4 in skeletal muscle. CONCLUSIONS: The present study demonstrates that IH can cause IR and reduced expression of GLUT4 in both mRNA and protein levels in skeletal muscle of rats. Rimonabant treatment can improve IH - induced IR, and the upregulation of GLUT4 expression may be involved in this process.
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Regulación de la Expresión Génica , Transportador de Glucosa de Tipo 4/metabolismo , Glucosa/metabolismo , Hipoxia , Músculo Esquelético/metabolismo , Piperidinas/uso terapéutico , Pirazoles/uso terapéutico , Animales , Glucemia/análisis , Antagonistas de Receptores de Cannabinoides/química , Inmunohistoquímica , Resistencia a la Insulina , Masculino , Músculo Esquelético/efectos de los fármacos , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Receptor Cannabinoide CB1/metabolismo , Rimonabant , Apnea Obstructiva del Sueño/tratamiento farmacológicoAsunto(s)
Proteínas Bacterianas/metabolismo , Regulación Bacteriana de la Expresión Génica , Regiones Promotoras Genéticas , Sinorhizobium meliloti/metabolismo , Factores de Transcripción/metabolismo , Proteínas Bacterianas/genética , Sinorhizobium meliloti/genética , Factores de Transcripción/genéticaAsunto(s)
Medicago sativa/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Nódulos de las Raíces de las Plantas/metabolismo , Estrés Fisiológico , Radicales Libres , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Mutación , Nitrógeno/química , Oxígeno/química , Proteínas de Plantas/metabolismo , Brotes de la PlantaRESUMEN
End-to-end weakly supervised semantic segmentation (E2E-WSSS) aims at optimizing a segmentation model in a single-stage training process based on only image annotations. Existing methods adopt an online-trained classification branch to provide pseudo annotations for supervising the segmentation branch. However, this strategy makes the classification branch dominate the whole concurrent training process, hindering these two branches from assisting each other. In our work, we treat these two branches equally by viewing them as diverse ways to generate the segmentation map, and add interactions on both their supervision and operation to achieve mutual promotion. For this purpose, a bidirectional supervision mechanism is elaborated to force the consistency between the outputs of these two branches. Thus, the segmentation branch can also give feedback to the classification branch to enhance the quality of localization seeds. Moreover, our method also designs interaction operations between these two branches to exchange their knowledge to assist each other. Experiments indicate our work outperforms existing end-to-end weakly supervised segmentation methods. Codes are available at https://github.com/zh460045050/BMP-WSSS.
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The abuse and overuse of antibiotics let drug-resistant bacteria emerges. Antibacterial photodynamic therapy (APDT) has shown outstanding merits to eliminate the drug-resistant bacteria via cytotoxic reactive oxygen species produced by irradiating photosensitizer. However, most of photosensitizers are not effective for Gram-negative bacteria elimination. Herein conjugates of NBS, a photosensitizer, linked with one (NBS-DPA-Zn) or two (NBS-2DPA-Zn) equivalents of zinc-dipicolylamine (Zn-DPA) have been designed to achieve the functional recognition of different bacteria. Due to the cationic character of NBS and metal transfer channel effect of Zn-DPA, NBS-DPA-Zn exhibited the first regent to distinguish P. aeruginosa from other Gram-negative bacteria. Whereas NBS-2DPA-Zn showed broad-spectrum antibacterial effect because the two arm of double Zn-DPA enhanced interactions with anionic membranes of bacteria, led the bacteria aggregation and thus provided the efficacy of APDT to bacteria and corresponding biofilm. In combination with a hydrogel of Pluronic, NBS-2DPA-Zn@gel shows promising clinical application in mixed bacterial diabetic mouse model infection. This might propose a new method that can realize functional identification and elimination of bacteria through intelligent regulation of Zn-DPA, and shows excellent potential for antibacterial application.
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Antibacterianos , Bacterias Gramnegativas , Fotoquimioterapia , Fármacos Fotosensibilizantes , Picolinas , Ácidos Picolínicos , Animales , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/farmacología , Fármacos Fotosensibilizantes/química , Antibacterianos/farmacología , Bacterias Gramnegativas/efectos de los fármacos , Ratones , Ácidos Picolínicos/química , Compuestos Organometálicos/química , Compuestos Organometálicos/farmacología , Biopelículas/efectos de los fármacos , Zinc/química , Pseudomonas aeruginosa/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Infecciones por Bacterias Gramnegativas/tratamiento farmacológicoRESUMEN
To successfully colonize legume root nodules, rhizobia must effectively evade host-generated reactive oxygen species (ROS). LsrB, a redox regulator from Sinorhizobium meliloti, is essential for symbiosis with alfalfa (Medicago sativa). The three cysteine residues in LsrB's substrate domain play distinct roles in activating downstream redox genes. The study found that LsrB's substrate-binding domain, dependent on the cysteine residue Cys146, is involved in oxidized glutathione (GSSG) resistance and alfalfa nodulation symbiosis. LsrB homologues from other rhizobia, with Cys172/Cys238 or Cys146, enhance GSSG resistance and complement lsrB mutant's symbiotic nodulation. Substituting amino acids in Azorhizobium caulinodans LsrB with Cys restores lsrB mutant phenotypes. The lsrB deletion mutant shows increased sensitivity to NCR247, suggesting an interaction with host plant-derived NCRs in alfalfa nodules. Our findings reveal that the key cysteine residue in the LsrB's substrate domain is vital for rhizobium-legume symbiosis.
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Proteínas Bacterianas , Glutatión , Medicago sativa , Nodulación de la Raíz de la Planta , Sinorhizobium meliloti , Simbiosis , Sinorhizobium meliloti/genética , Sinorhizobium meliloti/metabolismo , Sinorhizobium meliloti/fisiología , Medicago sativa/microbiología , Medicago sativa/metabolismo , Medicago sativa/genética , Medicago sativa/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Proteínas Bacterianas/química , Glutatión/metabolismo , Nodulación de la Raíz de la Planta/genética , Oxidación-Reducción , Cisteína/metabolismo , Cisteína/química , Nódulos de las Raíces de las Plantas/microbiología , Nódulos de las Raíces de las Plantas/metabolismo , Nódulos de las Raíces de las Plantas/genéticaRESUMEN
Purpose: To investigate the choroidal vascularity index (CVI) and choroidal structural changes in children with nephrotic syndrome. Methods: This was a cross-sectional study involving 45 children with primary nephrotic syndrome and 40 normal controls. All participants underwent enhanced depth imaging-optical coherence tomography examinations. An automatic segmentation method based on deep learning was used to segment the choroidal vessels and stroma, and the choroidal volume (CV), vascular volume (VV), and CVI within a 4.5 mm diameter circular area centered around the macular fovea were obtained. Clinical data, including blood lipids, serum proteins, renal function, and renal injury indicators, were collected from the patients. Results: Compared with normal controls, children with nephrotic syndrome had a significant increase in CV (nephrotic syndrome: 4.132 ± 0.464 vs. normal controls: 3.873 ± 0.574; P = 0.024); no significant change in VV (nephrotic syndrome: 1.276 ± 0.173 vs. normal controls: 1.277 ± 0.165; P = 0.971); and a significant decrease in the CVI (nephrotic syndrome: 0.308 [range, 0.270-0.386] vs. normal controls: 0.330 [range, 0.288-0.387]; P < 0.001). In the correlation analysis, the CVI was positively correlated with serum total protein, serum albumin, serum prealbumin, ratio of serum albumin to globulin, and 24-hour urine volume and was negatively correlated with total cholesterol, low-density lipoprotein cholesterol, urinary protein concentration, and ratio of urinary transferrin to creatinine (all P < 0.05). Conclusions: The CVI is significantly reduced in children with nephrotic syndrome, and the decrease in the CVI parallels the severity of kidney disease, indicating choroidal involvement in the process of nephrotic syndrome. Translational Relevance: Our findings contribute to a deeper understanding of how nephrotic syndrome affects the choroid.
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Síndrome Nefrótico , Niño , Humanos , Síndrome Nefrótico/complicaciones , Estudios Transversales , Coroides/diagnóstico por imagen , Fóvea Central , ColesterolRESUMEN
Background: The choroid is the most vascularized structure in the human eye, associated with numerous retinal and choroidal diseases. However, the vessel distribution of choroidal sublayers has yet to be effectively explored due to the lack of suitable tools for visualization and analysis. Methods: In this paper, we present a novel choroidal angiography strategy to more effectively evaluate vessels within choroidal sublayers in the clinic. Our approach utilizes a segmentation model to extract choroidal vessels from OCT B-scans layer by layer. Furthermore, we ensure that the model, trained on B-scans with high choroidal quality, can proficiently handle the low-quality B-scans commonly collected in clinical practice for reconstruction vessel distributions. By treating this process as a cross-domain segmentation task, we propose an ensemble discriminative mean teacher structure to address the specificities inherent in this cross-domain segmentation process. The proposed structure can select representative samples with minimal label noise for self-training and enhance the adaptation strength of adversarial training. Results: Experiments demonstrate the effectiveness of the proposed structure, achieving a dice score of 77.28 for choroidal vessel segmentation. This validates our strategy to provide satisfactory choroidal angiography noninvasively, supportting the analysis of choroidal vessel distribution for paitients with choroidal diseases. We observed that patients with central serous chorioretinopathy have evidently (P < 0.05) lower vascular indexes at all choroidal sublayers than healthy individuals, especially in the region beyond central fovea of macula (larger than 6 mm). Conclusions: We release the code and training set of the proposed method as the first noninvasive mechnism to assist clinical application for the analysis of choroidal vessels.
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China, is characterized by its remarkable ethnical diversity, which necessitates whole genome variation data from multiple populations as crucial tools for advancing population genetics and precision medical research. However, there has been a scarcity of research concentrating on the whole genome of ethnic minority groups. To fill this gap, we developed the Guizhou Multi-ethnic Genome Database (GMGD). It comprises whole genome sequencing data from 476 healthy unrelated individuals spanning 11 ethnic minorities groups in Guizhou Province, Southwest China, including Bouyei, Dong, Miao, Yi, Bai, Gelo, Zhuang, Tujia, Yao, Hui, and Sui. The GMGD database comprises more than 16.33 million variants in GRCh38 and 16.20 million variants in GRCh37. Among these, approximately 11.9% (1,956,322) of the variants in GRCh38 and 18.5% (3,009,431) of the variants in GRCh37 are entirely new and do not exist in the dbSNP database. These novel variants shed light on the genetic diversity landscape across these populations, providing valuable insights with an average coverage of 5.5 ×. This makes GMGD the largest genome-wide database encompassing the most diverse ethnic groups to date. The GMGD interactive interface facilitates researchers with multi-dimensional mutation search methods and displays population frequency differences among global populations. Furthermore, GMGD is equipped with a genotype-imputation function, enabling enhanced capabilities for low-depth genomic research or targeted region capture studies. GMGD offers unique insights into the genomic variation landscape of different ethnic groups, which are freely accessible at https://db.cngb.org/pop/gmgd/ .
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Pueblo Asiatico , Bases de Datos Genéticas , Etnicidad , Genoma Humano , Humanos , China/etnología , Etnicidad/genética , Variación Genética , Genética de Población/métodos , Grupos Minoritarios , Polimorfismo de Nucleótido Simple , Secuenciación Completa del Genoma/métodos , Pueblo Asiatico/genéticaRESUMEN
Repeated sevoflurane exposure in neonatal mice can leads to neuronal apoptosis and mitochondrial dysfunction. The mitochondria are responsible for energy production to maintain homeostasis in the central nervous system. The mitochondria-associated endoplasmic reticulum membrane (MAM) is located between the mitochondria and endoplasmic reticulum (ER), and it is critical for mitochondrial function and cell survival. MAM malfunction contributes to neurodegeneration, however, whether it is involved in sevoflurane-induced neurotoxicity remains unknown. Our study demonstrated that repeated sevoflurane exposure induced mitochondrial dysfunction and dampened the MAM structure. The upregulated ER-mitochondria tethering enhanced Ca2+ transition from the cytosol to the mitochondria. Overload of mitochondrial Ca2+ contributed to opening of the mitochondrial permeability transition pore (mPTP), which caused neuronal apoptosis. Mitofusin 2(Mfn2), a key regulator of ER-mitochondria contacts, was found to be suppressed after repeated sevoflurane exposure, while restoration of Mfn2 expression alleviated cognitive dysfunction due to repeated sevoflurane exposure in the adult mice. These evidences suggest that sevoflurane-induced MAM malfunction is vulnerable to Mfn2 suppression, and the enhanced ER-mitochondria contacts promotes mitochondrial Ca2+ overload, contributing to mPTP opening and neuronal apoptosis. This paper sheds light on a novel mechanism of sevoflurane-induced neurotoxicity. Furthermore, targeting Mfn2-mediated regulation of the MAM structure and mitochondrial function may provide a therapeutic advantage in sevoflurane-induced neurodegeneration.
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Retículo Endoplásmico , GTP Fosfohidrolasas , Mitocondrias , Sevoflurano , Animales , Sevoflurano/toxicidad , Sevoflurano/farmacología , GTP Fosfohidrolasas/metabolismo , Ratones , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/metabolismo , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Ratones Endogámicos C57BL , Apoptosis/efectos de los fármacos , Anestésicos por Inhalación/toxicidad , Anestésicos por Inhalación/farmacología , Masculino , Calcio/metabolismo , Membranas Intracelulares/efectos de los fármacos , Membranas Intracelulares/metabolismo , Proteínas de Transporte de Membrana Mitocondrial/metabolismo , Proteínas de Transporte de Membrana Mitocondrial/efectos de los fármacosRESUMEN
The enzyme φC31 integrase from Streptomyces phage has been documented as functional in mammalian cells and, therefore, has the potential to be a powerful gene manipulation tool. However, the activity of this enzyme is cell-type dependent. The more active mutant forms of φC31 integrase are required. Therefore, a rapid and effective method should be developed to detect the intracellular activity of φC31 integrase. We devised in this study an integrase-inversion cassette that contains the enhanced green fluorescent protein (EGFP) gene and the reverse complementary DsRed gene, which are flanked by attB and reverse complementary attP. This cassette can be inverted by φC31 integrase, thereby altering the fluorescent protein expression. Thus, φC31 integrase activity can be qualitatively or quantitatively evaluated based on the detected fluorescence. Furthermore, this cassette-based method was applied to several cell types, demonstrating that it is an efficient and reliable tool for measuring φC31 integrase activity in mammalian cells.
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Bacteriófagos/enzimología , Colorantes Fluorescentes/análisis , Proteínas Fluorescentes Verdes/análisis , Integrasas/metabolismo , Proteínas Luminiscentes/análisis , Streptomyces/virología , Animales , Línea Celular , Pruebas de Enzimas , Colorantes Fluorescentes/metabolismo , Expresión Génica , Proteínas Fluorescentes Verdes/genética , Humanos , Integrasas/genética , Proteínas Luminiscentes/genética , TransfecciónRESUMEN
Uncertainty, as the predominant characteristic of the contemporary landscape, poses significant challenges and exerts profound influence on individual decision making and behaviors; however, there remains a limited understanding of its impact on university student behavior. Building upon the uncertainty management theory, this study presents a conceptual framework to investigate the impact of perceived environmental uncertainty on university students' anxiety levels and behaviors, including academic engagement and prosocial behavior. Additionally, our model proposes that the intolerance of uncertainty moderates a mediating effect on anxiety. These hypotheses are empirically tested using a sample of 221 Chinese university students. The results reveal a positive relationship between perceived environmental uncertainty and anxiety among university students; subsequently, anxiety exerts a negative influence on both academic engagement and prosocial behavior. Furthermore, we find that anxiety serves as a psychological mediator between perceived environmental uncertainty and both academic engagement and prosocial behavior. This research also underscores the significance of the intolerance of uncertainty in shaping university students' involvement in academic pursuits when confronted with anxiety stemming from perceived environmental uncertainty. Consequently, these findings have practical implications for facilitating university students' adaptive coping strategies in uncertain contexts and mitigating the negative effects of anxiety on their behavioral responses.