Recent advances in early pregnancy loss diagnosis in dairy cows: New approaches.

Early pregnancy loss is a primary cause of low reproductive rates in dairy cows, posing severe economic losses to dairy farming. The accurate diagnosis of dairy cows with early pregnancy loss allows for oestrus synchronization, shortening day open, and increasing the overall conception rate of the herd. Several techniques are available for detecting early pregnancy loss in dairy cows, including rectal ultrasound, circulating blood progesterone, and pregnancy-associated glycoproteins (PAGs). Yet, there is a need to improve on existing techniques and develop novel strategies to identify cows with early pregnancy loss accurately. This manuscript reviews the applications of rectal ultrasound, circulating blood progesterone concentration, and PAGs in the diagnosis of pregnancy loss in dairy cows. The manuscript also discusses the recent progress of new technologies, including colour Doppler ultrasound (CDUS), interferon tau-induced genes (ISGs), and exosomal miRNA in diagnosing pregnancy loss in dairy cows. This study will provide an option for producers to re-breed cows with pregnancy loss, thereby reducing the calving interval and economic costs. Meanwhile, this manuscript might also act as a reference for exploring more economical and precise diagnostic technologies for early pregnancy loss in dairy cows.

Understanding AP2/ERF Transcription Factor Responses and Tolerance to Various Abiotic Stresses in Plants: A Comprehensive Review.

Abiotic stress is an adverse environmental factor that severely affects plant growth and development, and plants have developed complex regulatory mechanisms to adapt to these unfavourable conditions through long-term evolution. In recent years, many transcription factor families of genes have been identified to regulate the ability of plants to respond to abiotic stresses. Among them, the AP2/ERF (APETALA2/ethylene responsive factor) family is a large class of plant-specific proteins that regulate plant response to abiotic stresses and can also play a role in regulating plant growth and development. This paper reviews the structural features and classification of AP2/ERF transcription factors that are involved in transcriptional regulation, reciprocal proteins, downstream genes, and hormone-dependent signalling and hormone-independent signalling pathways in response to abiotic stress. The AP2/ERF transcription factors can synergise with hormone signalling to form cross-regulatory networks in response to and tolerance of abiotic stresses. Many of the AP2/ERF transcription factors activate the expression of abiotic stress-responsive genes that are dependent or independent of abscisic acid and ethylene in response to abscisic acid and ethylene. In addition, the AP2/ERF transcription factors are involved in gibberellin, auxin, brassinosteroid, and cytokinin-mediated abiotic stress responses. The study of AP2/ERF transcription factors and interacting proteins, as well as the identification of their downstream target genes, can provide us with a more comprehensive understanding of the mechanism of plant action in response to abiotic stress, which can improve plants' ability to tolerate abiotic stress and provide a more theoretical basis for increasing plant yield under abiotic stress.

Rfamide-related peptide-3(RFRP-3) receptor gene is expressed in mouse ovarian granulosa cells: Potential role of RFRP-3 in steroidogenesis and apoptosis.

RFRP-3 is a functional ortholog of avian GnIH and regulates reproductive activities in the gonads of animals. However, the role of RFRP-3 in the function of ovarian granulosa cells in mice remains unclear. First, we detected the expression of the RFRP-3 receptor (GPR147) in the ovarian granulosa cells of mice. Second, the effect of RFRP-3 treatment on estradiol and progesterone secretions from granulosa cells was tested by ELISA. Meanwhile, the expression of genes and proteins regulating steroid hormone synthesis was respectively examined by qPCR and western blot. Furthermore, the effect of RFRP-3 treatment on the apoptosis of granulosa cells was analyzed. The results revealed that the GPR147 protein (a RFRP-3 receptor) was expressed in the ovarian granulosa cells of mice. Low and medium doses RFRP-3 treatment significantly reduced progesterone secretion in the granulosa cells (P < 0.05), while RFRP-3 suppressed p450scc, 3ß-HSD, StAR, and FSHR expression in a non-dose-dependent manner. Moreover, RFRP-3 treatment might induce the apoptosis of granulosa cells. Additionally, low doses RFRP-3 significantly reduced p-ERK1/2 protein expression (P < 0.05) in the ovarian granulosa cells. We here, for the first time, confirmed that GPR147 was expressed in the ovarian granulosa cells of mice. Our findings suggested that and RFRP-3 regulates the granulosa cell function through the ERK signaling pathway, which will lay the foundation for uncovering molecular mechanisms by which RFRP-3 regulates follicle development in future.

Study on the Pattern of Postpartum Uterine Involution in Dairy Cows.

Postpartum uterine involution is necessary for the normal reproduction of dairy cows. The study aimed to investigate the pattern of postpartum uterine involution and the impact of parity on uterine involution in Chinese Holstein dairy cows. The diameter of the uterine cervix, pregnant uterine horn, and non-pregnant uterine horn were monitored using a B-mode veterinary ultrasound scanner at 5, 10, 15, 20, 25, 30, 35, and 40 days, respectively, after parturition in both multiparous and primiparous dairy cows. Meanwhile, the concentrations of hydroxyproline, E2, and IGF-1 were detected using ELISA at 5, 10, 15, 20, 30, and 40 d after parturition in both multiparous and primiparous dairy cows. Furthermore, the duration of uterine involution was compared in the multiparous and primiparous dairy cows. The results demonstrated that the diameter of the uterine cervix and the pregnant uterine horn did not decrease any further at 25 days postpartum for both the multiparous cows and the primiparous cows. Hydroxyproline levels gradually decreased with uterine involution; however, there was no significant variation in IGF-1 concentrations during uterine involution in the dairy cows. Although E2 concentrations of the peripheral plasma displayed an upward trend from day 5 to day 15 in the two groups of postpartum cows, there was no significant difference between the two groups during uterine involution. These results suggest that postpartum uterine involution was around 25 days postpartum in both the primiparous dairy cows and the multiparous Chinese Holstein dairy cows. Parity did not affect uterine involution in the postpartum Chinese Holstein dairy cows. The hydroxyproline levels of the peripheral blood may be an indicator of uterine involution in postpartum cows. Nonetheless, IGF-1 and E2 levels of the periphery blood are not associated with uterine involution in Chinese Holstein dairy cows.

MicroRNA: A Dynamic Player from Signalling to Abiotic Tolerance in Plants.

MicroRNAs (miRNAs) are a class of non-coding single-stranded RNA molecules composed of approximately 20-24 nucleotides in plants. They play an important regulatory role in plant growth and development and as a signal in abiotic tolerance. Some abiotic stresses include drought, salt, cold, high temperature, heavy metals and nutritional elements. miRNAs affect gene expression by manipulating the cleavage, translational expression or DNA methylation of target messenger RNAs (mRNAs). This review describes the current progress in the field considering two aspects: (i) the way miRNAs are produced and regulated and (ii) the way miRNA/target genes are used in plant responses to various abiotic stresses. Studying the molecular mechanism of action of miRNAs' downstream target genes could optimize the genetic manipulation of crop growth and development conditions to provide a more theoretically optimized basis for improving crop production. MicroRNA is a novel signalling mechanism in interplant communication relating to abiotic tolerance.

Exosomes: The role in mammalian reproductive regulation and pregnancy-related diseases.

Exosomes are a kind of extracellular vesicles that are produced and secreted by different mammalian cells. They serve as cargo proteins and can transfer different kinds of biomolecules, including proteins, lipids, and nucleic acids, which consequently act on target cells to exert different biological effects. Recent years have witnessed a significant increase in the number of studies on exosomes due to the potential effects of exosomes in the diagnosis and treatment of cancers, neurodegenerative diseases, and immune disorders. Previous studies have demonstrated that exosomal contents, especially miRNAs, are implicated in numerous physiological processes such as reproduction, and are crucial regulators of mammalian reproduction and pregnancy-related diseases. Here, we describe the origin, composition, and intercellular communication of exosomes, and discuss their functions in follicular development, early embryonic development, embryonic implantation, male reproduction and development of pregnancy-related diseases in humans and animals. We believe this study will provide a foundation for revealing the mechanism of exosomes in regulating mammalian reproduction, and providing new approaches and ideas for the diagnosis and treatment of pregnancy-related diseases.

Analysis of reproduction-related transcriptomes on pineal-hypothalamic-pituitary-ovarian tissues during estrus and anestrus in Tan sheep.

Seasonal estrus is an important factor limiting the fertility of some animals such as sheep. Promoting estrus in the anestrus season is one of the major ways in improving the fecundity of seasonally breeding animals. The pineal-hypothalamus-pituitary-ovary (PHPO) axis plays a decisive role in regulating animal reproduction. However, the molecular mechanisms by which the PHPO axis regulates seasonal reproduction in animals are not well understood, especially in Tan sheep. To this end, we collected pineal, hypothalamus, pituitary and ovary tissues from Tan sheep during estrus and anestrus for RNA-Sequencing, and performed bioinformatics analysis on the entire regulatory axis of the pineal-hypothalamic-pituitary-ovary (PHPO). The results showed that 940, 1,638, 750, and 971 DEGs (differentially expressed genes, DEGs) were identified in pineal, hypothalamus, pituitary and ovary, respectively. GO analysis showed that DEGs from PHPO axis-related tissues were mainly enriched in "biological processes" such as transmembrane transport, peptide and amide biosynthesis and DNA synthesis. Meanwhile, KEGG enrichment analysis showed that the bile acid secretion pathway and the neuroactive ligand-receptor interaction pathway were significantly enriched. Additionally, four potential candidate genes related to seasonal reproduction (VEGFA, CDC20, ASPM, and PLCG2) were identified by gene expression profiling and protein-protein interaction (PPI) analysis. These findings will contribute to be better understanding of seasonal reproduction regulation in Tan sheep and will serve as a useful reference for molecular breeding of high fertility Tan sheep.

Integrative Analysis of miRNA-mRNA in Ovarian Granulosa Cells Treated with Kisspeptin in Tan Sheep.

Kisspeptin is a peptide hormone encoded by the kiss-1 gene that regulates animal reproduction. Our studies revealed that kisspeptin can regulate steroid hormone production and promote cell proliferation in ovarian granulosa cells of Tan sheep, but the mechanism has not yet been fully understood. We speculated that kisspeptin might promote steroid hormone production and cell proliferation by mediating the expression of specific miRNA and mRNA in granulosa cells. Accordingly, after granulosa cells were treated with kisspeptin, the RNA of cells was extracted to construct a cDNA library, and miRNA-mRNA sequencing was performed. Results showed that 1303 expressed genes and 605 expressed miRNAs were identified. Furthermore, eight differentially expressed miRNAs were found, and their target genes were significantly enriched in progesterone synthesis/metabolism, hormone biosynthesis, ovulation cycle, and steroid metabolism regulation. Meanwhile, mRNA was significantly enriched in steroid biosynthesis, IL-17 signaling pathway, and GnRH signaling pathway. Integrative analysis of miRNA-mRNA revealed that the significantly different oar-let-7b targets eight genes, of which EGR1 (early growth response-1) might play a significant role in regulating the function of granulosa cells, and miR-10a regulates lipid metabolism and steroid hormone synthesis by targeting HNRNPD. Additionally, PPI analysis revealed genes that are not miRNA targets but crucial to other biological processes in granulosa cells, implying that kisspeptin may also indirectly regulate granulosa cell function by these pathways. The findings of this work may help understand the molecular mechanism of kisspeptin regulating steroid hormone secretion, cell proliferation, and other physiological functions in ovarian granulosa cells of Tan sheep.

OsDREB2B, an AP2/ERF transcription factor, negatively regulates plant height by conferring GA metabolism in rice.

The AP2/ERF family is a large group of plant-specific transcription factors that play an important role in many biological processes, such as growth, development, and abiotic stress responses. OsDREB2B, a dehydration responsive factor (DRE/CRT) in the DREB subgroup of the AP2/ERF family, is associated with abiotic stress responses, such as cold, drought, salt, and heat stress, in Arabidopsis or rice. However, its role in regulating plant growth and development in rice is unclear. In this study, we reported a new function of OsDREB2B, which negatively regulates plant height in rice. Compared with wild type (WT), OsDREB2B-overexpressing (OE) rice exhibited dwarf phenotypes, such as reduction in plant height, internode length, and seed length, as well as grain yield, while the knockout mutants developed by CRISPR/Cas9 technology exhibited similar phenotypes. Spatial expression analysis revealed that OsDREB2B was highly expressed in the leaf sheaths. Under exogenous GA3 application, OsDREB2B expression was induced, and the length of the second leaf sheath of the OsDREB2B-OE lines recovered to that of the WT. OsDREB2B localized to the nucleus of the rice protoplast acted as a transcription activator and upregulated OsAP2-39 by directly binding to its promoter. OsDREB2B-OE lines reduced endogenous bioactive GA levels by downregulating seven GA biosynthesis genes and upregulating eight GA deactivation genes but not GA signaling genes. The yeast two-hybrid assay and bimolecular fluorescence complementation assay showed that OsDREB2B interacted with OsWRKY21. In summary, our study suggests that OsDREB2B plays a negative role in rice growth and development by regulating GA metabolic gene expression, which is mediated by OsAP2-39 and OsWRKY21, thereby reducing GA content and rice plant height.

Physiological responses and transcriptome analysis of Spirodela polyrhiza under red, blue, and white light.

MAIN CONCLUSION: Red light (RL) accelerated starch accumulation in S. polyrhiza, but higher protein content under blue light (BL) was associated with the upregulation of most DEGs enriched for specific GO terms and KEGG pathways. Red light (RL) and blue light (BL) greatly influence the growth and physiological processes of duckweed. Physiological and molecular mechanisms underlying the response of duckweed to different light qualities remain unclear. This study employed physiological and transcriptomic analyses on duckweed, Spirodela polyrhiza "5510", to elucidate its differential response mechanisms under RL, BL, and white light conditions. Changes in growth indicators, ultrastructure alterations, metabolite accumulations, and differentially expressed genes (DEGs) were measured. The results showed that BL promoted both biomass and protein accumulations, while RL promoted starch accumulation. A total of 633, 518, and 985 DEGs were found in white-vs-red, white-vs-blue, and red-vs-blue comparison groups, respectively. In Gene Ontology (GO) enrichment analysis, the DEGs in all three comparison groups were significantly enriched in two GO terms, carboxylic acid metabolic process and lyase activity. In Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, the DEGs were greatly enriched in two pathways, histidine metabolism and isoquinoline alkaloid biosynthesis. Higher protein content under BL was associated with the upregulation of most DEGs enriched with the GO terms and KEGG pathways. Furthermore, the light qualities influenced the gene expression patterns of other metabolic pathways, like carotenoid biosynthesis, and the regulation of these genes may explain the level of photosynthetic pigment content. The results revealed the physiological changes and transcriptome-level responses of duckweed to three light qualities, thereby providing bases for further research studies on the ability of duckweed as a biomass energy source.

Involvement of rice transcription factor OsERF19 in response to ABA and salt stress responses.

Soil salinity is a major environmental stressor that restricts the growth and yield of crops. Plants have evolved more complicated and precise mechanisms to cope with salt stress, as they cannot escape from harmful environments. In the current study we identified and characterized an AP2/ERF transcription factor in rice, OsERF19. The expression of OsERF19 was slightly repressed by salt stress or abscisic acid (ABA) treatment. OsERF19-overexpression (OsERF19-OX) plants displayed enhanced tolerance to salt stress and ABA hypersensitivity compared to wild type and control plants. Furthermore, OsLEA3, OsNHX1, OsHKT6, and OsOTS1 were upregulated in OsERF19-OX plants when the plants were subjected to salt stress. OsRAB21, OsNCED5, and OsP5CS1 were also upregulated in OsERF19-OX plants treated with ABA. Yeast one-hybrid and dual luciferase reporter assays demonstrated that OsERF19 directly targets the promoters of OsOTS1 and OsNCED5 and further increases their transcription. These results suggest that the transcription factor OsERF19 plays a positive role in salt stress and ABA responses in rice.

OsGATA16, a GATA Transcription Factor, Confers Cold Tolerance by Repressing OsWRKY45-1 at the Seedling Stage in Rice.

BACKGROUND: Cold stress is the main abiotic stress in rice, which seriously affects the growth and yield of rice. Identification of cold tolerance genes is of great significance for rice to solve these problems. GATA-family transcription factors involve diverse biological functions, however, their role in cold tolerance in rice remains unclear. RESULTS: In this study, a GATA-type zinc finger transcription factor OsGATA16, which can improve cold tolerance, was isolated and characterized from rice. OsGATA16 belongs to OsGATA subfamily-II and contains 11 putative phosphorylation sites, a nuclear localization signal (NLS), and other several conserved domains. OsGATA16 was expressed in all plant tissues, with the strongest in panicles. It was induced by cold and ABA treatments, but was repressed by drought, cytokinin and JA, and acted as a transcriptional suppressor in the nucleus. Overexpression of OsGATA16 improves cold tolerance of rice at seedling stage. Under cold stress treatments, the transcription of four cold-related genes OsWRKY45-1, OsSRFP1, OsCYL4, and OsMYB30 was repressed in OsGATA16-overexpressing (OE) rice compared with wild-type (WT). Interestingly, OsGATA16 bound to the promoter of OsWRKY45-1 and repressed its expression. In addition, haplotype analysis showed that OsGATA16 polarized between the two major rice subspecies japonica and indica, and had a non-synonymous SNP8 (336G) associated with cold tolerance. CONCLUSION: OsGATA16 is a GATA transcription factor, which improves cold tolerance at seedling stage in rice. It acts as a positive regulator of cold tolerance by repressing some cold-related genes such as OsWRKY45-1, OsSRFP1, OsCYL4 and OsMYB30. Additionally, OsGATA16 has a non-synonymous SNP8 (336G) associated with cold tolerance on CDS region. This study provides a theoretical basis for elucidating the mechanism of cold tolerance in rice and new germplasm resources for rice breeding.

Rice Transcription Factor OsWRKY55 Is Involved in the Drought Response and Regulation of Plant Growth.

WRKY transcription factors (TFs) have been reported to respond to biotic and abiotic stresses and regulate plant growth and development. However, the molecular mechanisms of WRKY TFs involved in drought stress and regulating plant height in rice remain largely unknown. In this study, we found that transgenic rice lines overexpressing OsWRKY55 (OsWRKY55-OE) exhibited reduced drought resistance. The OsWRKY55-OE lines showed faster water loss and greater accumulation of hydrogen peroxide (H2O2) and superoxide radical (O2-·) compared to wild-type (WT) plants under drought conditions. OsWRKY55 was expressed in various tissues and was induced by drought and abscisic acid (ABA) treatments. Through yeast two-hybrid assays, we found that OsWRKY55 interacted with four mitogen-activated protein kinases (MAPKs) that could be induced by drought, including OsMPK7, OsMPK9, OsMPK20-1, and OsMPK20-4. The activation effects of the four OsMPKs on OsWRKY55 transcriptional activity were demonstrated by a GAL4-dependent chimeric transactivation assay in rice protoplasts. Furthermore, OsWRKY55 was able to reduce plant height under normal conditions by decreasing the cell size. In addition, based on a dual luciferase reporter assay, OsWRKY55 was shown to bind to the promoter of OsAP2-39 through a yeast one-hybrid assay and positively regulate OsAP2-39 expression. These results suggest that OsWRKY55 plays a critical role in responses to drought stress and the regulation of plant height in rice, further providing valuable information for crop improvement.

Influencing factors and growth kinetics analysis of carbon nanotube growth on the surface of continuous fibers.

Carbon nanotubes (CNTs) were continuously grown on the surface of the moving carbon fiber by chemical vapor deposition method using a custom-designed production line to prepare composite reinforcements on a large-scale. The systematic study of different parameters affecting the CNT growth revealed simple growth kinetics, which helps to control the surface morphology and structural quality of CNTs. Since hydrogen maintains the activity of the catalyst, it promotes the growth of CNTs in a continuous process. The increase of acetylene partial pressure promotes the accumulation of amorphous or graphite carbon on the catalyst surface, resulting in the decrease of CNT growth rate when acetylene concentration reaches 40%. The growth temperature significantly affects the CNT diameter and structural quality. As the temperature increases, the crystallinity of the tube wall increases obviously, and the CNT diameter increases due to the aggregate growth of the catalyst particles. According to the Arrhenius formula, the apparent activation energy is observed to be 0.67 eV, which proves that both bulk diffusion and surface diffusion exist when activated carbon passes through the catalyst to form CNTs.

Exploring the trade-offs between electric heating policy and carbon mitigation in China.

China has enacted a series of policies since 2015 to substitute electricity for in-home combustion for rural residential heating. The Electric Heating Policy (EHP) has contributed to significant improvements in air quality, benefiting hundreds of millions of people. This shift, however, has resulted in a sharp increase in electric loads and associated carbon emissions. Here, we show that China's EHP will greatly increase carbon emissions. We develop a theoretical model to quantify the carbon emissions from power generation and rural residential heating sectors. We found that in 2015, an additional 101.69-162.89 megatons of CO2 could potentially be emitted if EHP was implemented in 45-55% of rural residents in Northern China. In 2020, the incremental carbon emission is expected to reach 130.03-197.87 megatons. Fortunately, the growth of carbon emission will slow down due to China's urbanization progress. In 2030, the carbon emission increase induced by EHP will drop to 119.19-177.47 megatons. Finally, we conclude two kinds of practical pathways toward low-carbon electric heating, and provide techno-economic analyses.

A Novel AP2/ERF Transcription Factor, OsRPH1, Negatively Regulates Plant Height in Rice.

The APETALA 2/ethylene response factors (AP2/ERF) are widespread in the plant kingdom and play essential roles in regulating plant growth and development as well as defense responses. In this study, a novel rice AP2/ERF transcription factor gene, OsRPH1, was isolated and functionally characterized. OsRPH1 falls into group-IVa of the AP2/ERF family. OsRPH1 protein was found to be localized in the nucleus and possessed transcriptional activity. Overexpression of OsRPH1 resulted in a decrease in plant height and length of internode and leaf sheath as well as other abnormal characters in rice. The length of the second leaf sheath of OsRPH1-overexpressing (OE) plants recovered to that of Kitaake (non-transgenic recipient) in response to exogenous gibberellin A3 (GA3) application. The expression of GA biosynthesis genes (OsGA20ox1-OsGA20ox4, OsGA3ox1, and OsGA3ox2) was significantly downregulated, whereas that of GA inactivation genes (OsGA2ox7, OsGA2ox9, and OsGA2ox10) was significantly upregulated in OsRPH1-OE plants. Endogenous bioactive GA contents significantly decreased in OsRPH1-OE plants. OsRPH1 interacted with a blue light receptor, OsCRY1b, in a blue light-dependent manner. Taken together, our results demonstrate that OsRPH1 negatively regulates plant height and bioactive GA content by controlling the expression of GA metabolism genes in rice. OsRPH1 is involved in blue light inhibition of leaf sheath elongation by interacting with OsCRY1b.