The newly identified liver-enriched gene 1 (LEG1) encodes a protein with a characteristic domain of unknown function 781 (DUF781/LEG1), constituting a protein family with only one member in mammals. A functional study in zebrafish suggested that LEG1 genes are involved in liver development, while the platypus LEG1 homolog, Monotreme Lactation Protein (MLP), which is enriched in the mammary gland and milk, acts as an antibacterial substance. However, no functional studies on eutherian LEG1s have been published to date. Thus, we here report the first functional prediction study at the cellular level. As previously reported, eutherian LEG1s can be classified into three paralogous groups. Pigs have all three LEG1 genes (pLEG1s), while humans and mice have retained only LEG1a. Hence, pLEG1s might represent an ideal model for studying LEG1 gene functions. RNA-seq was performed by the overexpression of pLEG1s and platypus MLP in HepG2 cells. Enrichment analysis showed that pLEG1a and pLEG1b might exhibit little function in liver cells; however, pLEG1c is probably involved in the endoplasmic reticulum (ER) stress response and protein folding. Additionally, gene set enrichment analysis revealed that platypus MLP shows antibacterial activity, confirming the functional study in platypus. Therefore, our study showed from the transcriptomic perspective that mammalian LEG1s have different functions in liver cells due to the subfunctionalization of paralogous genes.
Weak tissue adhesion remains a major challenge in clinical translation of microneedle patches. Mimicking the structural features of honeybee stingers, stiff polymeric microneedles with unidirectionally backward facing barbs were fabricated and embedded into various elastomer films to produce self-interlocking microneedle patches. The spirality of the barbing pattern was adjusted to increase interlocking efficiency. In addition, the micro-bleeding caused by microneedle puncturing adhered the porous surface of the patch substrate to the target tissue via coagulation. In the demonstrative application of myocardial infarction treatment, the bioinspired microneedle patches firmly fixed on challenging beating hearts, significantly reduced cardiac wall stress and strain in the infarct, maintained left ventricular function and morphology. In addition, the microneedle patch was minimally invasively implanted onto beating porcine heart in 10 minutes, free of sutures and adhesives. Therefore, the honeybee stinger inspired microneedles could provide an adaptive and convenient means to adhere patches for various medical applications.Funding Information: This study is financially supported by the National Key Research and Development Program of China (No. 2019YFE0117400), National Natural Science Foundation of China (No. 32000971, 52005097, 81941003), "Leading Goose" R&D Program of Zhejiang, Natural Science Foundation of Shanghai (No. 21ZR1401300).Conflict of Interests: Y.Z., Y.L., C.G., T.R., L.S., X.H. have patent applications (CN113907915A) related to the present study and thus may have related financial interests. All other authors declare no competing interests.Ethical Approval: All animal experiments were approved by the Institutional Animal Care and Use Committee of Zhejiang University (No. ZJU20210245).
Porcine reproductive and respiratory syndrome (PRRS), which is caused by the PRRS virus (PRRSV), has resulted in large economic losses for the swine industry. The virus has shown remarkable genetic diversity since its discovery. In our study, we investigated mutation types in the evolution of PRRSV for both in vivo and in vitro passaging of the virus. Sequence alignment analysis demonstrated that the most common hypermutations expressed were A→G/U→C and G→A/C→U. The data provide a new theoretical basis for PRRSV evolution.Le syndrome reproducteur et respiratoire porcin (SRRP), qui est causé par le virus SRRP (VSRRP), a entrainé de grosses pertes économiques à l’industrie porcine. Le virus a démontré une remarquable diversité génétique depuis sa mise en évidence. Dans notre étude, nous avons examiné les types de mutation dans l’évolution du VSRRP lors du passage in vivo et in vitro du virus. L’analyse d’alignement des séquences a démontré que les hypermutations les plus fréquemment exprimées étaient A→G/U→C et G→A/C→U. Ces données fournissent une nouvelle base théorique pour l’évolution du VSRRP.(Traduit par Docteur Serge Messier).
Current understandings of the initiation of the trophectoderm (TE) program during mammalian embryonic development lack evidence of how TE-associated factors such as GATA3 and CDX2 participate in bovine lineage specification. In this study, we describe effects of TE-associated factors on the expression of lineage specification marker genes such as SOX2, OCT4, NANOG, GATA6 and SOX17, by using cytosine base editor system. We successfully knockout GATA3 or CDX2 in bovine embryos with robust efficiency. However, GATA3 or CDX2 deletion does not affect the developmental potential of embryos to reach blastocyst stage. Interestingly, GATA3 deletion downregulates the NANOG expression in bovine blastocysts. Further analysis of the mosaic embryos shows that GATA3 is required for NANOG in the TE of bovine blastocysts. Single blastocyst RNA-seq analysis reveals that GATA3 deletion disrupts the transcriptome in bovine blastocysts. Altogether, we propose that GATA3 plays an important role in maintaining TE lineage program in bovine embryos and the functional role of GATA3 is species-specific.
Magnetic resonance imaging (MRI) is a popular imaging tool that is valuable for the early detection and monitoring of malignancies because it does not involve radiation and is noninvasive. Metal-based contrast agents (CAs) are commonly used in clinical settings despite concerns about the toxicity of free metals. Therefore, finding alternative nontoxic imaging probes is vital. In this work, we have synthesized and effectively utilized sustainable biomass lignin-based all-organic nanoconjugates linked with nitroxide radicals as MRI CAs. Lignin grafted with poly(4-glycidyloxy-2,2,6,6-tetramethylpiperidine-1-oxyl) (LPGT) exhibits a longitudinal relaxivity of 0.54 mM
Pulmonary arterial hypertension (PAH) is a rare cardiovascular disease with very high mortality rate. The currently available therapeutic strategies, which improve symptoms, cannot fundamentally reverse the condition. Thus, new therapeutic strategies need to be established. Our research analyzed three microarray datasets of lung tissues from human PAH samples retrieved from the Gene Expression Omnibus (GEO) database. We combined two datasets for subsequent analyses, with the batch effects removed. In the merged dataset, 542 DEGs were identified and the key module relevant to PAH was selected using WGCNA. GO and KEGG analyses of DEGs and the key module indicated that the pre-ribosome, ribosome biogenesis, centriole, ATPase activity, helicase activity, hypertrophic cardiomyopathy, melanoma, and dilated cardiomyopathy pathways are involved in PAH. With the filtering standard (|MM| > 0.95 and |GS| > 0.90), 70 hub genes were identified. Subsequently, five candidate marker genes (CDC5L, AP3B1, ZFYVE16, DDX46, and PHAX) in the key module were found through overlapping with the top thirty genes calculated by two different methods in CytoHubb. Two of them (CDC5L and DDX46) were found to be significantly upregulated both in the merged dataset and the validating dataset in PAH patients. Meanwhile, expression of the selected genes in lung from PAH chicken measured by qRT-PCR and the ROC curve analyses further verified the potential marker genes' predictive value for PAH. In conclusion, CDC5L and DDX46 may be marker genes and potential therapeutic targets for PAH.
Diagnostic and therapeutic illumination on internal organs and tissues with high controllability and adaptability in terms of spectrum, area, depth, and intensity remains a major challenge. Here, we present a flexible, biodegradable photonic device called iCarP with a micrometer scale air gap between a refractive polyester patch and the embedded removable tapered optical fiber. ICarP combines the advantages of light diffraction by the tapered optical fiber, dual refractions in the air gap, and reflection inside the patch to obtain a bulb-like illumination, guiding light towards target tissue. We show that iCarP achieves large area, high intensity, wide spectrum, continuous or pulsatile, deeply penetrating illumination without puncturing the target tissues and demonstrate that it supports phototherapies with different photosensitizers. We find that the photonic device is compatible with thoracoscopy-based minimally invasive implantation onto beating hearts. These initial results show that iCarP could be a safe, precise and widely applicable device suitable for internal organs and tissue illumination and associated diagnosis and therapy.
During mammalian preimplantation development, the transition from morula to blastocyst is a critical biological event. This process involves polarization and initial specification of lineages, regulated by various transcription factors that have been extensively studied in mice. Our single-cell RNA sequencing analyses revealed that TEAD3 is specifically expressed in the trophectoderm cells of bovine preimplantation embryos, unlike in mice. The objective of this study is to determine the functional role of TEAD3 in bovine preimplantation development. While TEAD3 knockdown does not affect blastocyst formation in cattle, embryos fail to progress to the blastocyst stage when both TEAD3 and TEAD4, another member of the TEAD family, are disrupted using RNA interference and base editing techniques, respectively. This finding suggests a redundant role for TEAD3 and TEAD4 in preimplantation development in cattle. RNA sequencing analysis identified dysregulation of 215 genes, with 53 genes upregulated and 162 genes downregulated. Notably, we observed a reduction in the expression of trophectoderm-specifier genes KRT8, KRT18, and EZR, as well as HIPPO signaling pathway components. Immunofluorescence analysis further revealed that the protein expression levels of KRT8 and EZR were significantly decreased. Importantly, the initial expression of trophectoderm lineage-specific factors such as TFAP2C and GATA3, as well as the inner cell mass lineage-specific transcription factor OCT4, remained unaffected. This contrasts with the role of TEAD4 in directly regulating trophectoderm lineage specification in mice. Thus, our studies demonstrate that TEAD3 and TEAD4 play essential and redundant roles upstream of TE fate decisions during preimplantation development in cattle.
The emergence of the first three lineages during development is orchestrated by a network of transcription factors, which are best characterized in mice. However, the role and regulation of these factors are not completely conserved in other mammals, including human and cattle. Here, we establish a gene inactivation system with a robust efficiency by introducing premature codon with cytosine base editors in bovine early embryos. By using this approach, we have determined the functional consequences of three critical lineage-specific genes (SOX2, OCT4 and CDX2) in bovine embryos. In particular, SOX2 knockout results in a failure of the establishment of pluripotency in blastocysts. Indeed, OCT4 level is significantly reduced and NANOG barely detectable. Furthermore, the formation of primitive endoderm is compromised with few SOX17 positive cells. RNA-seq analysis of single blastocysts (day 7.5) reveals dysregulation of 2074 genes, among which 90% are up-regulated in SOX2-null blastocysts. Intriguingly, more than a dozen lineage-specific genes, including OCT4 and NANOG, are down-regulated. Moreover, SOX2 level is sustained in the trophectoderm in absence of CDX2. However, OCT4 knockout does not affect the expression of SOX2. Overall, we propose that SOX2 is indispensable for OCT4 and NANOG expression and CDX2 represses the expression of SOX2 in the trophectoderm in cattle, which are all in sharp contrast with results in mice.
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