image-recognition-parameter-analysis-concrete-vibration-state-based

Image Recognition and Parameter Analysis of Concrete Vibration State Based on Support Vector Machine

In this paper, the directional gradient histogram technology is employed to extract the features of concrete image samples captured under different ...

generation-of-maternal-mutants-using-zpccas9-knock-in-zebrafish

Generation of Maternal Mutants Using <em>zpc:cas9</em> Knock-in Zebrafish

Oogenesis and early embryonic development are critically dependent on maternally derived mRNAs and proteins. Eliminating these maternal factors ...

Wuhan Construction Engineering Group Co. Ltd.

Shandong University

Supermolecular hydrogels were prepared by α-cyclodeatrin (α-CD) and Tyloxapol, which can be considered as an oligomer of the nonionic surfactant polyoxyethylene tert-octylphenyl ether (TX-100) with a polymerization degree below 7. Two carbon materials, graphene oxide (GO) and graphene, were mixed into the α-CD/Tyloxapol hydrogel to adjust the physicochemical properties of hydrogel. In order to get stable graphene dispersion and then mix it with α-CD/Tyloxapol hydrogel, both TX-100 and Tyloxapol were used to disperse graphene for comparison. Interestingly, it can be found that TX-100 could disperse graphene better than Tyloxapol owing to smaller molecular size of TX-100 compared with Tyloxapol. Then, both the α-CD/Tyloxapol/GO and α-CD/Tyloxapol/graphene hydrogels were characterized by transmission electron microscopy (TEM), field emission scanning electron microscopy (FE-SEM), Fourier transform infrared (FT-IR) spectroscopy, small angle X-ray scattering (SAXS), X-ray diffraction (XRD) and rheological measurements. The results revealed that the addition of carbon materials into α-CD/Tyloxapol hydrogel can change their microstructures and the rheological properties. Furthermore, it can be confirmed that a little amount of carbon materials could induce fluorescence quenching sharply which could be a promising candidate for optical sensor.

Manipulation the properties of supramolecular hydrogels of α-cyclodextrin/Tyloxapol/carbon-based nanomaterials.

The spontaneous formation of hybrid submicrometer spheres that were composed by a Weakley-type polyoxometalate Na[EuWO]·32HO (denoted as EuW) and cationic peptide (K8) through a simple ionic self-assembly method was investigated. The approach presented in this study is an extended research which combined a biomolecule and a functional inorganic polyoxoanion for the fabrication of a multifunctional material. The K8/EuW hybrid submicrometer spheres were fully characterized by transmission electron microscopy, field-emission scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, confocal laser scanning microscopy, and fluorescence spectra. The results indicated that the electrostatic interaction, hydrogen-bonding interaction, and combined hydrophobic interaction between EuW and K8 favored the formation of the smooth submicrometer sphere structure. Once the EuW/K8 submicrometer spheres were forming, the fluorescence of EuW was reduced due to the hydrogen bonding between the ammonium group of K8 and the oxygen atom of EuW that blocked the hopping of the d electron in EuW. Interestingly, our submicrometer spheres showed excellent decomposition efficiency toward organic pollutants such as the dye of methylene blue (MB), suggesting their promising applications in the treatment of wastewater.

Self-Assembly of Peptide-Polyoxometalate Hybrid Sub-Micrometer Spheres for Photocatalytic Degradation of Methylene Blue.

Circular RNA has been revealed as a potential biomarker in multiple malignancies. However, few studies have focused on its potential to be prognostic markers in lung squamous cell carcinoma (LSCC). In this work, we aimed to build a prognostic model of resected LSCC based on circular RNA pyruvate dehydrogenase kinase 1 (circPDK1) and other clinicopathological factors.

Prognostic model based on circular RNA circPDK1 for resected lung squamous cell carcinoma.

Lens transparency is established by abundant accumulation of crystallin proteins and loss of organelles in the fiber cells. It requires an efficient translation of lens messenger RNAs (mRNAs) to overcome the progressively reduced transcriptional activity that results from denucleation. Inappropriate regulation of this process impairs lens differentiation and causes cataract formation. However, the regulatory mechanism promoting protein synthesis from lens-expressed mRNAs remains unclear. Here we show that in zebrafish, the RNA-binding protein Rbm24 is critically required for the accumulation of crystallin proteins and terminal differentiation of lens fiber cells. In the developing lens, Rbm24 binds to a wide spectrum of lens-specific mRNAs through the RNA recognition motif and interacts with cytoplasmic polyadenylation element-binding protein (Cpeb1b) and cytoplasmic poly(A)-binding protein (Pabpc1l) through the C-terminal region. Loss of Rbm24 reduces the stability of a subset of lens mRNAs encoding heat shock proteins and shortens the poly(A) tail length of  mRNAs encoding lens structural components, thereby preventing their translation into functional proteins. This severely impairs lens transparency and results in blindness. Consistent with its highly conserved expression in differentiating lens fiber cells, the findings suggest that vertebrate Rbm24 represents a key regulator of cytoplasmic polyadenylation and plays an essential role in the posttranscriptional control of lens development.

Rbm24 controls poly(A) tail length and translation efficiency of  mRNAs in the lens via cytoplasmic polyadenylation.

Syne2b/nesprin-2 is a giant protein implicated in tethering the nucleus to the cytoskeleton and plays an important role in maintaining cellular architecture. Epiboly is a conserved morphogenetic movement that involves extensive spreading and thinning of the epithelial blastoderm to shape the embryo and organize the three germ layers. Dynamic cytoskeletal organization is critical for this process, but how it is regulated remains elusive. Here we generated a zebrafish  mutant line and analyzed the effects of impaired Syne2b function during early development. By CRISPR/Cas9-mediated genome editing, we obtained a large deletion in the  locus, predicted to cause truncation of the nuclear localization KASH domain in the translated protein. Maternal and zygotic  embryos showed delayed epiboly initiation and progression without defects in embryonic patterning. Remarkably, disruption of Syne2b function severely impaired cytoskeletal organization across the embryo, leading to aberrant clustering of F-actin at multiple cell contact regions and abnormal cell shape changes. These caused disintegration of the epithelial blastoderm before the end of gastrulation in most severely affected embryos. Moreover, the migration of yolk nuclear syncytium also became defective, likely due to disorganized cytoskeletal networks at the blastoderm margin and in the yolk cell. These findings demonstrate an essential function of Syne2b in maintaining cytoskeletal architecture and epithelial integrity during epiboly movement.

Syne2b/Nesprin-2 Is Required for Actin Organization and Epithelial Integrity During Epiboly Movement in Zebrafish.

Maternal products are exclusive factors to drive oogenesis and early embryonic development. As disrupting maternal gene functions is either time-consuming or technically challenging, early developmental programs regulated by maternal factors remain mostly elusive. We provide a transgenic approach to inactivate maternal genes in zebrafish primary oocytes. By introducing three tandem single guide RNA (sgRNA) expression cassettes and a green fluorescent protein (GFP) reporter into Tg(:) embryos, we efficiently obtained maternal  and  mutants among GFP-positive F offspring. Notably, most of these maternal mutants displayed either sgRNA site-spanning genomic deletions or unintended large deletions extending distantly from the sgRNA targets, suggesting a prominent deletion-prone tendency of genome editing in the oocyte. Thus, our method allows maternal gene knockout in the absence of viable and fertile homozygous mutant adults. This approach is particularly time-saving and can be applied for functional screening of maternal factors and generating genomic deletions in zebrafish.

Rapid generation of maternal mutants via oocyte transgenic expression of CRISPR-Cas9 and sgRNAs in zebrafish.

Maternal products are those mRNAs and proteins deposited during oogenesis, which play critical roles in controlling oocyte formation, fertilization, and early embryonic development. However, loss-of-function studies for these maternal factors are still lacking, mainly because of the prolonged period of transgenerational screening and technical barriers that prevent the generation of maternal (M) and maternal and zygotic (MZ) mutant embryos. By the transgenic expression of multiple sgRNAs targeting a single gene of interest in the background of a transgenic line Tg(:) with oocyte-specific  expression, we have successfully obtained maternal or maternal-zygotic mutant for single genes in F1 embryos. In this work, we tandemly connected a maternal GFP marker and eight sgRNA expression units to target  and  simultaneously and introduced this construct to the genome of Tg(:) by meganuclease I- I. As expected, we confirmed the existence of M;M embryos with strong defective convergence and extension movement during gastrulation among outcrossed GFP positive F1 offspring. The MZ;MZ embryos were also obtained by crossing the mutant carrying mosaic F0 female with ; male fish. This proof-of-principle thus highlights the potential of this conditional knockout strategy to circumvent the current difficulty in the study of genes with multiple functionally redundant paralogs.

A Time-Saving Strategy to Generate Double Maternal Mutants by an Oocyte-Specific Conditional Knockout System in Zebrafish.

Use of zebrafish as animal model for various diseases during early developmental stages has been exponentially increased with the aim to achieve the best representative results in this transparent fish. Recent studies documented that Rbm24a mutant causes cataract formation and resulted in blindness using the zebrafish model. Therefore, correct interpretation of studies that aimed for molecular approaches, a description of comparative and in-depth analysis of development of lens in wildtype and mutant is crucial to obtain the correct conclusion. In this study, we use a gold standard method the Transmission Electron Microscopy (TEM) to analysis the lens development in rbm24a mutant zebrafish. Firstly, we compare the cellular structures at 16-20 h post fertilization (hpf), the lens placode in ectoderm indicated delay lens development in rbm24a mutant than wildtype (siblings) zebrafish. At 33 hpf, loosely appeared lens fiber cells showed heterogenous electron density with numbers of mitochondria in lens of rbm24a mutant, revealed the influence of gene mutation in lens development. A detail ultrastructure of lens of rbm24a mutant also presented at 33 hpf. Comparatively in wildtype (siblings) at 33 hpf, lens exhibited homogenous electron density in tightly packed lens fiber cells with few mitochondria. Furthermore, to characterize the lens in rbm24a mutant we obtained data of cellular structures on 25 hpf and 1.5 days' post fertilization (dpf). At 25 hpf in mutant zebrafish, the detached solid sphere lens mass from ectoderm showed karyorrhexis, mitophagy and vesicles (also multivesicular bodies), these cellular structures supposed to hamper the development of future fiber cells. Moreover, at 1.5 dpf in mutant, nuclear excisosome, multilamellar bodies and irregular shaped mitochondria in heterogenous electron dense cytoplasm of lens fiber cells, collectively shown affected lens transparency. In summary the ultrastructure results of lens of rbm24a mutant zebrafish expand our knowledge and give reflection of different cellular activities like autophagy, apoptosis, vesicles (multivesicular bodies) and nuclear excisosomes which play their role in transparency achievement.

Cellular activity of autophagy and multivesicular bodies in lens fiber cells during early lens development in rbm24a mutant of zebrafish: Ultrastructure analysis.

Evidence has shown a strong relationship between smoking and epithelial mesenchymal transition (EMT). α5-nicotinic acetylcholine receptor (α5-nAChR) contributes to nicotine-induced lung cancer cell EMT. The cytoskeleton-associated protein PLEK2 is mainly involved in cytoskeletal protein recombination and cell stretch migration regulation, which is closely related to EMT. However, little is known about the link between nicotine/α5-nAChR and PLEK2 in lung adenocarcinoma (LUAD). Here, we identified a link between α5-nAChR and PLEK2 in LUAD. α5-nAChR expression was correlated with PLEK2 expression, smoking status and lower survival in vivo. α5-nAChR mediated nicotine-induced PLEK2 expression via STAT3. α5-nAChR/PLEK2 signaling is involved in LUAD cell migration, invasion and stemness. Moreover, PLEK2 was found to interact with CFL1 in nicotine-induced EMT in LUAD cells. Furthermore, the functional link among α5-nAChR, PLEK2 and CFL1 was confirmed in mouse xenograft tissues and human LUAD tissues. These findings reveal a novel α5-nAChR/PLEK2/CFL1 pathway involved in nicotine-induced LUAD progression.

PLEK2 mediates metastasis and invasion via α5-nAChR activation in nicotine-induced lung adenocarcinoma.

Primary cilia are enriched in signaling receptors, and defects in their formation or function can induce conditions such as polycystic kidney disease, postaxial hexadactyly, and microphthalmia. Mammalian Hedgehog (Hh) signaling is important in the development of primary cilia, and TMEM216, a transmembrane protein that localizes to the base of cilia, is also implicated in ciliogenesis in zebrafish. Here, we found that -deficient mice had impaired Hh signaling and displayed typical ciliopathic phenotypes. These phenomena were also observed in cells deficient in TMEM216. Furthermore, TMEM216 interacted with core Hh signaling proteins, including SUFU, a negative regulator of Hh, and GLI2/GLI3, transcription factors downstream of Hh. The competition between TMEM216 and SUFU for binding to GLI2/GLI3 inhibited the cleavage of GLI2/GLI3 into their repressor forms, which resulted in the nuclear accumulation of full-length GLI2 and the decreased nuclear localization of cleaved GLI3, ultimately leading to the activation of Hh signaling. Together, these data suggest that the TMEM216-SUFU-GLI2/GLI3 axis plays a role in TMEM216 deficiency-induced ciliopathies and Hh signaling abnormalities.

TMEM216 promotes primary ciliogenesis and Hedgehog signaling through the SUFU-GLI2/GLI3 axis.

Maternal mRNAs and proteins are produced during oogenesis by more than 60% of zebrafish genes. They are indispensable for fertilization and early embryogenesis. Generation and analysis of the maternal mutant is the most direct way to characterize the maternal function of the specific gene. However, due to the lethality of zygotic mutants, the maternal function of most genes in zebrafish remains elusive. Several methods have been developed to circumvent this obstacle, including mRNA rescue, germ-line replacement, oocyte microinjection in situ, mosaic mutation, and bacterial artificial chromosome (BAC)-mediated conditional rescue. Here, we provide an alternative approach to generate zebrafish maternal mutants rapidly and efficiently by introducing four tandem sgRNA expression cassettes into Tg() embryos. This method is more technically feasible and cost- and time-effective than other established methods. Key features • This protocol can circumvent the lethality or infertility of the zygotic mutants to obtain maternal mutants of the target gene. • This protocol is time-saving (one fish generation). • Using this protocol, double-gene maternal mutants can be obtained in a single generation. • Stable lines can be established to continuously produce maternal mutant embryos for the gene of interest.

Generation of Zebrafish Maternal Mutants via Oocyte-Specific Knockout System.

Double-stranded RNAs (dsRNAs), known as conserved pathogen-associated molecular patterns, activate the integrated stress response via interferon-induced protein kinase R (PKR), leading to global translation inhibition. However, the interferon system is inactive in pluripotent cells, leaving the mechanisms of dsRNA sensing and translational control unclear. In this study, we utilized early zebrafish embryos as a model of pluripotent cells and discovered a PKR-independent blockage of translation initiation by dsRNA stimulation. Prkra dimer was identified as the genuine dsRNA sensor. Upon dsRNA binding, the dimerized dsRNA-binding domain 3 of Prkra becomes activated to sequester the eIF2 complexes from the translation machinery, inhibiting global protein synthesis. This distinctive embryonic stress response restricts RNA virus replication in zebrafish embryos, is conserved in mouse embryonic stem cells, and compensates PKR function in differentiated cells. Therefore, the Prkra-mediated dsRNA sensing and translation control may serve as a common strategy for cells to adapt to environmental stresses.

Prkra dimer senses double-stranded RNAs to dictate global translation efficiency.

The germ granules are ribonucleoprotein (RNP) biomolecular condensates that determine the fate of primordial germ cells (PGCs) and serve as a model for studying RNP granule assembly. Here, we show that the maternal RNA-binding protein Rbm24a is a key factor governing the specific sorting of mRNAs into germ granules. Mechanistically, Rbm24a interacts with the germ plasm component Buc to dictate the specific recruitment of germ plasm mRNAs into phase-separated condensates. Germ plasm particles lacking Rbm24a and mRNAs fail to undergo kinesin-dependent transport toward cleavage furrows where small granules fuse into large aggregates. Therefore, the loss of maternal Rbm24a causes a complete degradation of the germ plasm and the disappearance of PGCs. These findings demonstrate that the Rbm24a/Buc complex functions as a nucleating organizer of germ granules, highlighting an emerging mechanism for RNA-binding proteins in reading and recruiting RNA components into a phase-separated protein scaffold.

Rbm24a dictates mRNA recruitment for germ granule assembly in zebrafish.

Our goal was to investigate the connection between asthma and HEI-2020.

ArticleTotal : 2	Year
Publication title Cited by 1	2024
Publication title	2025

Article	Year
Manipulation the properties of supramolecular hydrogels of α-cyclodextrin/Tyloxapol/carbon-based nanomaterials. Journal of colloid and interface science\| PubMed ID: 26828277	2016
Self-Assembly of Peptide-Polyoxometalate Hybrid Sub-Micrometer Spheres for Photocatalytic Degradation of Methylene Blue. The journal of physical chemistry. B\| PubMed ID: 29095615	2017
Prognostic model based on circular RNA circPDK1 for resected lung squamous cell carcinoma. Translational lung cancer research\| PubMed ID: 32010569	2019
Rbm24 controls poly(A) tail length and translation efficiency of mRNAs in the lens via cytoplasmic polyadenylation. Proceedings of the National Academy of Sciences of the United States of America\| PubMed ID: 32170011	2020
Syne2b/Nesprin-2 Is Required for Actin Organization and Epithelial Integrity During Epiboly Movement in Zebrafish. Frontiers in cell and developmental biology\| PubMed ID: 34222245	2021
Rapid generation of maternal mutants via oocyte transgenic expression of CRISPR-Cas9 and sgRNAs in zebrafish. Science advances\| PubMed ID: 34362733	2021
A Time-Saving Strategy to Generate Double Maternal Mutants by an Oocyte-Specific Conditional Knockout System in Zebrafish. Biology\| PubMed ID: 34440009	2021
Cellular activity of autophagy and multivesicular bodies in lens fiber cells during early lens development in rbm24a mutant of zebrafish: Ultrastructure analysis. Micron (Oxford, England : 1993)\| PubMed ID: 36965272	2023
PLEK2 mediates metastasis and invasion via α5-nAChR activation in nicotine-induced lung adenocarcinoma. Molecular carcinogenesis\| PubMed ID: 37921560	2024
TMEM216 promotes primary ciliogenesis and Hedgehog signaling through the SUFU-GLI2/GLI3 axis. Science signaling\| PubMed ID: 38261656	2024
Generation of Zebrafish Maternal Mutants via Oocyte-Specific Knockout System. Bio-protocol\| PubMed ID: 39525965	2024
Prkra dimer senses double-stranded RNAs to dictate global translation efficiency. Molecular cell\| PubMed ID: 40280134	2025
Rbm24a dictates mRNA recruitment for germ granule assembly in zebrafish. The EMBO journal\| PubMed ID: 40281355	2025
Association of the 2020 Healthy Eating Index with asthma in children and adolescents in the United States: a cross-sectional study of NHANES. The Journal of asthma : official journal of the Association for the Care of Asthma\| PubMed ID: 40439092	2025

Tong Lu

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