Mammalian interspecific hybrids provide unique advantages for mechanistic studies of speciation, gene expression regulation, and X chromosome inactivation (XCI) but are constrained by their limited ...natural resources. Previous artificially generated mammalian interspecific hybrid cells are usually tetraploids with unstable genomes and limited developmental abilities. Here, we report the generation of mouse-rat allodiploid embryonic stem cells (AdESCs) by fusing haploid ESCs of the two species. The AdESCs have a stable allodiploid genome and are capable of differentiating into all three germ layers and early-stage germ cells. Both the mouse and rat alleles have comparable contributions to the expression of most genes. We have proven AdESCs as a powerful tool to study the mechanisms regulating X chromosome inactivation and to identify X inactivation-escaping genes, as well as to efficiently identify genes regulating phenotypic differences between species. A similar method could be used to create hybrid AdESCs of other distantly related species.
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•The mouse-rat AdESCs are genetically stable and pluripotent•The AdESCs exhibit mid-parent and species-biased gene expression patterns•Mouse-specific X inactivation in allodiploid cells reveals Xi-escaping genes•The AdESCs provide a tool to study the species-specific pluripotency maintenance
The mouse-rat allodiploid ESCs generated by fusion of haploid ESCs exhibit intriguing gene expression patterns and mouse-specific X chromosome inactivation after differentiation, thereby providing a valuable tool for X chromosome inactivation and gene function studies.
Dysregulated expression of microRNAs (miRNAs or miRs) has been implicated in the pathophysiology of type 2 diabetes mellitus (T2DM). However, their underlying role in the complication of detrusor ...fibrosis remains poorly understood. Therefore, this study aimed to examine the potential functional relevance of miR-363 in detrusor fibrosis of rats with streptozotocin (STZ)-induced T2DM through the predicted target gene collagen type I alpha 2 (Col1a2). Immunohistochemical analysis found an increase in the positive expression of collagen type III alpha 1 (Col3a1) and Col1a2 in detrusor tissues, where miR-363 expression was decreased. Next, gain- and loss-of-function experiments were performed to clarify the effects of miR-363 and Col1a2 on the activities of bladder detrusor cells. Of note, binding affinity between miR-363 and Col1a2 was verified by a dual-luciferase reporter gene assay and RNA immunoprecipitation (RIP) assay. Upregulated miR-363 inhibited Col1a2 expression, which led to increased expression of B-cell lymphoma 2 (Bcl-2) and Smad7 and accelerated cell viability, along with decreases in cell apoptosis and Col3a1, Bcl-2-associated X protein (Bax), transforming growth factor (TGF)-β1, and Smad4 expressions. In conclusion, miR-363 upregulation reduces detrusor fibrosis in rats with STZ-induced T2DM through suppression of the TGF-β1/Smad signaling pathway by targeting Col1a2. Therefore, our study provided further insights for the development of new therapeutic targets for T2DM.
miR-363 can inhibit Col1a2, which promotes viability and reduces apoptosis of detrusor cells in rats with STZ-induced T2DM through the inhibition of the TGF-β1/Smad signaling pathway, which ultimately leads to the alleviation of detrusor fibrosis. This study provides new evidence regarding the functional actions of miR-363 in T2DM.
Wearable electronics, such as sensors, actuators, and supercapacitors, have attracted broad interest owing to their promising applications. Nevertheless, practical problems involving their ...sensitivity and stretchability remain as challenges. In this work, efforts were devoted to fabricating a highly stretchable and sensitive strain sensor based on dip-coating of graphene onto an electrospun thermoplastic polyurethane (TPU) nanofibrous membrane, followed by spinning of the TPU/graphene nanomembrane into an intertwined-coil configuration. Owing to the intertwined-coil configuration and the synergy of the two structures (nanoscale fiber gap and microscale twisting of the fiber gap), the conductive strain sensor showed a stretchability of 1100%. The self-inter-locking of the sensor prevents the coils from uncoiling. Thanks to the intertwined-coil configuration, most of the fibers were wrapped into the coils in the configuration, thus avoiding the falling off of graphene. This special configuration also endowed our strain sensor with an ability of recovery under a strain of 400%, which is higher than the stretching limit of knees and elbows in human motion. The strain sensor detected not only subtle movements (such as perceiving a pulse and identifying spoken words), but also large movements (such as recognizing the motion of fingers, wrists, knees, etc.), showing promising application potential to perform as flexible strain sensors.
Abstract
Background
Age at natural menopause (ANM) is an important index for women’s health. Either early or late ANM is associated with a series of adverse outcomes later in life. Despite being an ...inheritable trait, its genetic determinant has not yet been fully understood.
Methods
Aiming to better characterize the genetic architecture of ANM, we conducted genome-wide association study (GWAS) meta-analyses in European-specific as well as trans-ancestry samples by using GWAS summary statistics from the following 3 large studies: the Reproductive Genetics Consortium (ReproGen; N = 69 626), the UK Biobank cohort (UKBB; N = 111 593) and the BioBank Japan Project (BBJ; N = 43 861), followed by a series of bioinformatical assessments and functional annotations.
Results
By integrating the summary statistics from the 3 GWAS of up to 225 200 participants, this largest meta-analysis identified 49 novel loci and 3 secondary signals that were associated with ANM at the genome-wide significance level (P < 5 × 10−8). No population specificity or heterogeneity was observed at most of the associated loci. Functional annotations prioritized 90 candidate genes at the newly identified loci. Among the 26 traits that were genetically correlated with ANM, hormone replacement therapy (HRT) exerted a causal relationship, implying a causal pattern by which HRT was determined by ANM.
Conclusion
Our findings improved our understanding of the etiology of female menopause, as well as shed light on potential new therapies for abnormal menopause.
The study aims to investigate the analgesic effects of microRNA‐129‐5p (miR‐129‐5p) on bone cancer pain (BCP) by targeting Eph receptor B1 (EphB1) through the EphB1/EphrinB2 signaling pathway. BCP ...mice models were established, and C3H/HeJ female mice were classified into the normal, blank, negative control (NC), miR‐129‐5p mimics, miR‐129‐5p inhibitors, EphB1 knockout (KO), and miR‐129‐5p inhibitors + EphB1 KO groups. Quantitative reverse transcription polymerase chain reaction and Western blot analysis were used to evaluate the miR‐129‐5p expression, and messenger RNA (mRNA) and protein expressions of EphB1, p‐EphB1, EphrinB2, and p‐EphrinB2. EphB1 and EphrinB2 were highly activated in the tibias of BCP mice 7 days after the operation. EphB1 is a target gene of miR‐129‐5p. The mechanical withdrawal threshold increased in the miR‐129‐5p mimics, EphB1 KO and miR‐129‐5p inhibitors + EphB1 KO groups, but decreased in the miR‐129‐5p inhibitors group. Compared with the blank and the NC groups, the expression of miR‐129‐5p was significantly increased in the miR‐129‐5p mimics group, and the mRNA and protein expressions of EphrinB2, p‐EphrinB2, EphB1, and p‐EphB1 were significantly decreased, while in the miR‐129‐5p inhibitors group, the results were opposite (all
P < 0.05); the mRNA and protein expressions of EphrinB2, p‐EphrinB2, EphB1, and p‐EphB1 were significantly decreased in the EphB1 KO group (all
P < 0.05); the expression of miR‐129‐5p was significantly decreased in the miR‐129‐5p inhibitors + EphB1 KO group (
P < 0.05), while the mRNA and protein expressions of EphrinB2 and p‐EphrinB2 were not significantly different (
P > 0.05). The results indicated that upregulated miR‐129‐5p alleviate BCP via downregulation of the EphB1/EphrinB2 signaling pathway.
The results indicated that upregulated microRNA‐129‐5p alleviates bone cancer pain via downregulation of the Eph receptor B1/EphrinB2 signaling pathway.
The arsenic in livestock wastewater would induce adverse impact on the biological treatment technology such as anaerobic ammonium oxidation (anammox) process. Extracellular polymeric substances (EPS) ...play an important role in resisting such toxicity. Unfortunately, the role of EPS in protecting anammox from As(III) and the mechanisms underlying the protection still remains unclear. This work comprehensively evaluated the acute toxicity of arsenic on anammox sludge and investigated the binding property and interaction mechanism. The results revealed that the half maximal inhibitory concentration (IC50) of As(III) on anammox sludge was estimated to be 408 mg L−1, which decreased to 41.97 mg L−1 when EPS was exfoliated. Complexation and hydrophobic interactions were the leading forces in preventing arsenic invasion. Protein was the main component that complexes with As(III), and O–H, –NH, –CO were binding sites. The response sequence of organic component in EPS to As(III) was ordered as hydrocarbons-proteins-polysaccharides-aliphatic amines.
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•The IC50 of As(III) was estimated as 408 mg L−1 in anammox process.•EPS was able to construct buffer zones to prevent As(III) invasion.•Protein is the main component within EPS to complex with As(III).•Response sequence of functional groups to As(III) was clarified via spectra analysis.
Leaves are one of the vegetative organs of plants that are essential for plant growth and development.
(
) gene is an indoleacetic acid (IAA) transporter that plays a critical role in leaf ...development. To determine the function of
in leaf polarity formation in
, the transgenic lines with
overexpression (OE) and
reduced expression (RE) were analyzed using the
-mediated method. The RE lines displayed the characteristics of leaf margin adaxial upward curling, with lower expression of
resulting in greater rolling. Tissue localization of IAA in the auxin
reporter system proved that auxin in the RE was mainly distributed in the secondary veins, palisade tissues, and epidermal cells in the leaf margin area. The auxin content in the leaf margin area was significantly greater than that in the main vein tissue. The cell density of the palisade tissue and the ratio of palisade tissue to spongy tissue in the curled leaf margin of the RE lines were found to be significantly decreased. RNA-seq analysis revealed that the RE hormone-signaling pathway genes were significantly enriched compared with those of the OE and WT lines; in particular, the auxin response-related genes SAURs (i.e.,
,
,
, and
) and
were found to be significantly upregulated. qRT-PCR analysis indicated that
expression at the leaf margin was significantly lower than that near the main vein in the RE lines. In contrast, the expression levels of SAURs and
were significantly higher than those near the midrib. In conclusion,
regulates the expression of auxin response-related genes and the polar transport of auxin to change the polar form of the proximal and distal axes of birch leaves.
In this study, we tested three batches of HPV trivalent vaccines that were prepared in the pilot plant and stored in the prefilled syringes at 2 to 8°C. The tests of stability include identity, ...appearance, the volume of fill, pH, osmolarity, aluminum content, polysorbate-80 content, sterility test, bacterial endotoxin, abnormal toxicity, in vitro relative potency analysis, and in vivo potency evaluation. According to the results, the pH was slightly increased, the polysorbate-80 content was decreased slowly, and ED50 values of HPV16, HPV18, and HPV58 were slightly elevated compared with day 0. According to the tests above, the storage condition of the HPV trivalent vaccine is determined to be 2 to 8°C avoiding light.
The recent success of derivation of mammalian haploid embryonic stem cells (haESCs) has provided a powerful tool for large-scale functional analysis of the mammalian genome. However, haESCs rapidly ...become diploidized after differentiation, posing challenges for genetic analysis. Here, we show that the spontaneous diploidization of haESCs happens in metaphase due to mitotic slippage. Diploidization can be suppressed by small-molecule-mediated inhibition of CDK1 and ROCK. Through ROCK inhibition, we can generate haploid somatic cells of all three germ layers from haESCs, including terminally differentiated neurons. Using piggyBac transposon-based insertional mutagenesis, we generated a haploid neural cell library harboring genome-wide mutations for genetic screening. As a proof of concept, we screened for Mn2+-mediated toxicity and identified the Park2 gene. Our findings expand the applications of mouse haploid cell technology to somatic cell types and may also shed light on the mechanisms of ploidy maintenance.
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•Mitotic slippage causes spontaneous diploidization of mouse haESCs•Inhibition of CDK1 and ROCK suppresses the diploidization of mouse haESCs•ROCK inhibition can generate haploid somatic cells of all three germ layers•Haploid neural stem-cell-like cells can be used for genome-wide genetic screening
He et al. show that CDK1 and ROCK inhibition can suppress the spontaneous diploidization of haploid embryonic stem cells (haESCs) and generate haploid somatic cells of all three germ layers for genome-wide genetic screening.
Abstract
Background
Ferroptosis plays an essential role in lipopolysaccharide (LPS)-induced acute lung injury (ALI). Meteorin-like/Meteorin-β (Metrnβ) is a protein secreted by skeletal muscle and ...adipose tissue and plays a role in cardiovascular diseases. However, its role in acute lung injury has not been elucidated.
Methods
In this study, we used an adenovirus (Ad) delivery system to overexpress or knockdown Metrnβ in lung tissue to examine the role of Metrnβ in LPS-induced acute lung injury.
Results
We found that ferroptosis was increased during LPS-induced ALI. The expression of Metrnβ was reduced in ALI lung tissue. Overexpression of Metrnβ in lung tissue alleviated LPS-induced lung injury, inflammation, and ferroptosis. Moreover, Metrnβ knockout in lung tissue accelerated LPS-induced ALI, inflammation, and ferroptosis. We also cultured MLE-12 cells and transfected the cells with Ad-Metrnβ or Metrnβ siRNA. Metrnβ overexpression ameliorated LPS-induced MLE cell death, inflammation and ferroptosis, while Metrnβ knockdown aggregated cell survival and decreased inflammation and ferroptosis. Moreover, we found that Metrnβ enhanced ferroptosis-related Gpx4 expression and reduced ferroportin and ferritin levels. Furthermore, we found that Metrnβ positively regulated SIRT1 transcription thus inhibited P53, increased SLC7A11 expression. When we used the ferroptosis inhibitor ferrostatin-1, the deteriorating effects of Metrnβ knockout were abolished in ALI mice. Moreover, SIRT1 knockout also abolished the protective effects of Metrnβ overexpression in vivo.
Conclusions
Taken together, Metrnβ could protect LPS-induced ALI by activating SIRT1-P53- SLC7A11 mediated ferroptosis inhibition.
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