Exploring the relationships between hormone receptor, HER2 status, and bone involvement in the first distant metastases of in Chinese breast cancer patients who lacked HER2 targeted therapy.

BACKGROUND: This retrospective study explored the relationship between hormone receptor (HR), human epidermal growth factor receptor 2 (HER2) status, and bone involvement in the first distant metastases (DM) of Chinese breast cancer (BC) patients who lacked the HER2 targeted therapy. Such therapy was rarely received due to its lag approval or high cost in China compared with the developed countries. METHODS: All eligible women with primary unilateral stage I - III BC and first DM diagnosed in 2008-2018 at one cancer center were identified for enrollment. Based on chart records, a full or no/partial compliance status of endocrine therapy (ET) was assigned for HR-positive patients. Multivariate logistic regression was used to estimate the adjusted odd ratio (aOR), its 95%CI and p value. RESULTS: Four hundred eighteen patients had an average age of 50.7 years and median disease-free survival of 27.1 months at DM. Bone, lung, liver and brain metastasis rates in patients were 55.7%, 34.7%, 33.0% and 8.1%, respectively. Compared to HR-negative patients, HR-positive patients with the full and non/partial compliance of ET were significantly associated with higher risk of bone involvement with an aOR of 2.329 (1.316 - 1.741, p = 0.004) and 2.317 (1.330 - 4.036, p = 0.003), respectively. No difference of such risk was found between the two groups of ET compliance (p = 0.984) nor between HER2-negative and HER2-positive patients (aOR 0.827, p = 0.431). Stratified analyses further indicated that HR-positive was associated with bone involvement only in HER2-negative BC patients (p = 0.006-0.015). CONCLUSIONS: HR-positive tumors are significantly associated with bone involvement in HER2-negative metastatic BC patients. ET does not appear to impact this association. HER2 status per se is not associated with such risk.

Effect of different dry matter content on fermentation characteristics and nutritional quality of Napier grass silage with novel lactic acid bacteria strains.

To investigate the characteristics of different LAB strains isolated from subtropics and their effects on Napier grass (Pennisetum purpureum Schum.) silage with two dry matter (DM) levels, sugar fermentation pattern, and growth profiles of three screened lactic acid bacteria (LAB) strains [Pediococcus pentosaceus (PP04), Weissella cibaria (WC10), and Lactobacillus plantarum (LP694)] were characterized, and then used either individually or in combination at 1.0 × 106 cfu g-1 fresh weight to inoculate grass having 15% or 25% DM. Treatments were applied: (1) no inoculant (control); (2) PP04; (3) WC10; (4) LP694; (5) M-1 (PP04: WC10 = 2:1); (6) M-2 (PP04: LP694 = 1:2); (7) M-3 (WC10: LP694 = 2:1); (8) M-4 (PP04: WC10: LP694 = 2:1:1). The results showed that all inoculations increased LAB, DM recovery, and lactic acid (LA) concentration, while decreasing pH, the ammonia nitrogen/total nitrogen (NH3-N/TN), and butyric acid (BA) concentration compared to control group in both DM. However, the effect of inoculations was very limited at 15% DM. Silages with inoculants achieved higher silage quality at 25% DM than 15% DM. The different LAB inoculants result in significant differences in silage quality, while W. cibaria decreased the pH and inhibited the growth of undesirable bacteria and those characteristics were not affected by the DM content.

Curcumin ameliorates Staphylococcus aureus-induced mastitis injury through attenuating TLR2-mediated NF-κB activation.

PROBLEM: It has long been known that Staphylococcus aureus (S. aureus, serotype CP8) is associated with clinical mastitis in cows, and recent scientific studies have shown that curcumin (CUR) is effective in anti-inflammatory. However, the mechanism of action of curcumin on S. aureus-induced cows has not been fully understood. Therefore, this study investigated whether curcumin could improve the inflammation response in mice mastitis and to clarify the possible mechanism. METHOD: of study: A mouse mastitis model was established. The mice were administered curcumin (125 mg/kg), ciprofloxacin (130 mg/kg, CIP), and water (model group) for 5 days. RESULTS: CUR and CIP treatment prevented the S. aureus-induced mouse mastitis increase the levels of IL-2, IL-10, and IFN-γ and decrease levels of IL-6, IL-8, and TNF-α. Additionally, RT-PCR results showed that 20 µg/mL curcumin inhibited the mRNA expression of TNF-α, IL-6, IL-1ß, TRAF6 and MEKK1 in murine mammary epithelial cells (MMECs). Likewise, Western blotting results showed that CUR inhibited the expression of TRAF6 and MEKK1. CONCLUSION: These results indicated that CUR is superior to CIP in the prevention of mastitis, and the mechanism may be that the curative effect of CUR inhibits TLR-2 mediated NF-κB signaling pathway in mouse mastitis.

Metabolites of stable fly reduce diarrhea in mice by modulating the immune system, antioxidants, and composition of gut microbiota.

Escherichia coli (E. coli) O1-induced diarrhea is associated with intestinal microbial imbalance, however, the results of using oral antibiotics still remain poor. To overcome such problem, our study investigates the role of metabolites from stable flies (MSF) in the occurrence of diarrhea. The amino acid composition and molecular weight analysis of MSF by RP-HPLC and GPC, respectively. Besides the normal control group, SPF mice in other group were inoculated with E. coli O1 received treatment as follows over a period of 7 days saline solution (E. coli control), ciprofloxacin (0.13 g/kg; positive control) and MSF (2, 4 and 8 mg/kg) dosage. Throughout the experiment, defecation and body weights were examined and recorded. On the eighth day, after administering anesthesia, blood, tissue of small intestine samples were obtained for immunological and anti-oxidant. Small intestinal tissues and cecum contents samples were used for histopathological and 16S rDNA sequencing analysis. Our showed that MSF was rich in isoleucine, and its molecular weight less than 400 Da is 60.03%. MSF (4 and 8 mg/kg) and ciprofloxacin, significantly decreased IL-6, IL-8 and TNF-α levels, whereas, increased IL-2, IL-4, IL-10, INF-γ, IgA and IgG levels in mice having diarrhea. These treatments also reversed intestinal flora imbalance as indicated by the increased in Firmicutes-to-Bacteroidetes ratio and Clostridium levels (P < 0.05) and improved 5-HT, CAT and SOD levels. MSF favored diarrhea management as compared to ciprofloxacin, suggesting that MSF can be used in the management of E. coli O1-induced diarrhea, in normal gut microbiota and normal intestinal antioxidant function.

Regulation of Hedgehog signaling Offers A Novel Perspective for Bone Homeostasis Disorder Treatment.

The hedgehog (HH) signaling pathway is central to the regulation of bone development and homeostasis. HH signaling is not only involved in osteoblast differentiation from bone marrow mesenchymal stem cells (BM-MSCs), but also acts upstream within osteoblasts via the OPG/RANK/RANKL axis to control the expression of RANKL. HH signaling has been found to up-regulate parathyroid hormone related protein (PTHrP) expression in osteoblasts, which in turn activates its downstream targets nuclear factor of activated T cells (NFAT) and cAMP responsive element binding protein (CREB), and as a result CREB and NFAT cooperatively increase RANKL expression and osteoclastogenesis. Osteoblasts must remain in balance with osteoclasts in order to avoid excessive bone formation or resorption, thereby maintaining bone homeostasis. This review systemically summarizes the mechanisms whereby HH signaling induces osteoblast development and controls RANKL expression through PTHrP in osteoblasts. Proper targeting of HH signaling may offer a therapeutic option for treating bone homeostasis disorders.

Effect of nisin on microbiome-brain-gut axis neurochemicals by Escherichia coli-induced diarrhea in mice.

The effects of nisin on the neurochemicals, Aquaporin-3 (AQP-3) and intestinal microorganisms in the brain-gut axis of mice were analyzed by using enzyme linked immune sorbent assay (ELISA) and high throughput sequencing in this investigation, to further revealed the relationship between intestinal flora abundance in mice and neurochemicals in the brain-gut axis. Using HE staining found damage of structure of small intestine villi in the model group (Escherichia coli O1, E. coli O1). Compared with normal control and ciprofloxacin groups, using ELISA showed that nisin increased the highest norepinephrine (NE) expression in the brain, expression of 5-hydroxytryptamine (5-HT) and dopamine (DA) in the duodenum, and increased the expression of AQP-3 in jejunum. Using high-throughput sequencing showed the highest diversity of cecal microflora in nisin group (ACE-index = 1417.25, Chao1-index = 1378.45), but the cecal microflora in the negative control group (ACE-index = 969.54, Chao1-index = 340.29) exhibited the lowest species diversity. Our data indicated that nisin regulates neurochemicals, AQP-3 and cecal microflora imbalance in mice.

Genome-wide characterization of TCP family and their potential roles in abiotic stress resistance of oat (Avena sativa L.).

The TCP gene family members play multiple functions in plant growth and development and were named after the first three family members found in this family, TB1 (TEOSINTE BRANCHED 1), CYCLOIDEA (CYC), and Proliferating Cell Factor 1/2 (PCF1/2). Nitrogen (N) is a crucial element for forage yield; however, over-application of N fertilizer can increase agricultural production costs and environmental stress. Therefore, the discovery of low N tolerance genes is essential for the genetic improvement of superior oat germplasm and ecological protection. Oat (Avena sativa L.), is one of the world's staple grass forages, but no genome-wide analysis of TCP genes and their roles in low-nitrogen stress has been performed. This study identified the oat TCP gene family members using bioinformatics techniques. It analyzed their phylogeny, gene structure analysis, and expression patterns. The results showed that the AsTCP gene family includes 49 members, and most of the AsTCP-encoded proteins are neutral or acidic proteins; the phylogenetic tree classified the AsTCP gene family members into three subfamilies, and each subfamily has different conserved structural domains and functions. In addition, multiple cis-acting elements were detected in the promoter of the AsTCP genes, which were associated with abiotic stress, light response, and hormone response. The 49 AsTCP genes identified from oat were unevenly distributed on 18 oat chromosomes. The results of real-time quantitative polymerase chain reaction (qRT-PCR) showed that the AsTCP genes had different expression levels in various tissues under low nitrogen stress, which indicated that these genes (such as AsTCP01, AsTCP03, AsTCP22, and AsTCP38) played multiple roles in the growth and development of oat. In conclusion, this study analyzed the AsTCP gene family and their potential functions in low nitrogen stress at the genome-wide level, which lays a foundation for further analysis of the functions of AsTCP genes in oat and provides a theoretical basis for the exploration of excellent stress tolerance genes in oat. This study provides an essential basis for future in-depth studies of the TCP gene family in other oat genera and reveals new research ideas to improve gene utilization.

Occurrence characteristics and risk assessment of microplastics in agricultural soils in the loess hilly gully area of Yan' an, China.

Large amounts of microplastics accumulated in the soil of agricultural fields with the rapid development of mulch agriculture. The enrichment of microplastics endangered the growth of crops and food security, and it also posed ecological risks. In this study, we investigated microplastics in a typical agriculture area of Yan' an City, in the loess hilly gully area of China. The characteristics of microplastics including their abundances, sizes, and types were measured through laser direct infrared spectrometer. The potential sources of microplastics were analyzed and the risk of soil microplastic pollution was evaluated. It was elaborated that the average abundances of microplastics in soil, water, and fertilizer were 4505 ± 435 n·kg-1, 91 ± 27 n·L-1, and 39,629 ± 10,114 n·kg-1, respectively. Microplastics with particle sizes < 100 µm accounted for >90 %. The smaller the particle size, the higher the content of microplastics. The top three polymers were polyethylene (PE, 37.4 %), polyethylene terephthalate (PET, 15.0 %), and ethylene vinyl acetate (EVA, 8.9 %), respectively. Agricultural mulch, plastic film, domestic waste, surface water irrigation, and organic compost were probably the potential sources of soil microplastics. The ecological risk evaluation showed that overall sampling sites had a minor ecological risk of microplastic pollution based on their abundance, while the polymer type showed a relatively high ecological risk for the investigated agricultural soils. Polyvinylchloride (PVC) and polymethylmethacrylate (PMMA) contribute considerably to the ecological risk, and their inputs to the farmland environment should be strictly limited. There was no significant carcinogenic risk to humans. This study would provide the basic reference for the current situation and risk assessment of farmland soil microplastics pollution in the loess hilly gully area of China.

Study on the relationship between regional soil desertification and salinization and groundwater based on remote sensing inversion: A case study of the windy beach area in Northern Shaanxi.

Soil desertification and salinization are important environmental concerns in arid regions, and their relationship with groundwater change must be further clarified. However, the relationships among soil desertification, salinization, and groundwater are difficult to investigate on a large spatiotemporal scale using traditional ground surveys. In the windy beach area in Northern Shaanxi (WBANS), desertification and salinization problems coexist; therefore, this area was selected as the study area. The feasibility of implementing large-scale remote sensing inversions to identify the degree of desertification and salinization was verified based on measured data, and the degree of influence of groundwater burial depth (GBD) on desertification and salinization was quantified using the geodetector and residual trend analysis methods. The results showed that the GBD in the WBANS presented an increasing trend and the degree of salinization showed a decreasing trend. Moreover, the joint influence of the unique natural environment and anthropogenic activities has led to increases in fractional vegetation cover and considerable improvements in the ecological environment. The intensity of desertification explained by GBD in the WBANS increased significantly (p < 0.01) at a rate of change of 0.0190/year, with high q-values above 0.66 for both Yuyang and Shenmu. The contribution rate of potential evapotranspiration and precipitation to salinization in Yuyang and Shenmu was >97 %, and the contribution rate of GBD to salinization in Dingbian, Jingbian, and Hengshan was 34.78 %, 31.15 %, and 29.41 %, respectively. Overall, the suitable GBD in the WBANS is 2-4 m. The study results provide a reference for research on the inversion, monitoring, and prevention of desertification and salinization dynamics on a large spatiotemporal scale and offer a scientific basis for rationally determining GBD.

LiveBoost: A GB-based prediction system for liver fibrosis in chronic hepatitis B patients in China - A multi-center retrospective study.

Background: The aim of this study was to evaluate the accuracy of LiveBoost™, a gradient boosting (GB)-based prediction system based on standard biochemical values (AST, ALT, platelet count) and age, in Chinese patients with chronic hepatitis B (CHB) and compare its performance with FIB-4 (fibrosis-4 score) and APRI (the aspartate transaminase to platelet ratio index). Methods: This retrospective trial enrolled 454 participants, including 279 CHB patients who underwent liver biopsy and 175 normal controls from 3 centers in China. All participants underwent laboratory blood testing. LiveBoost was constructed using GB and FIB-4 and APRI were calculated from laboratory data. Results: LiveBoost outperformed APRI and FIB-4 in predicting hepatic fibrosis and cirrhosis. The GB model had an AUROC of 0.977 for CHB diagnosis, 0.804 for early and advanced fibrosis, and 0.836 for non-cirrhosis and cirrhosis, compared to AUROC of 0.554, 0.673 and 0.720 for FIB-4, AUROC of 0.977, 0.652 and 0.654 for APRI. Conclusions: LiveBoost is a more reliable and cost-effective method than APRI and FIB-4 for assessing liver fibrosis in Chinese patients with CHB.

[Characteristics and source of soil condensation water in Guanzhong Plain, China]. / å ³ä¸­å¹³åŽŸåœŸå£¤å‡ç»“æ°´é‡åŠæ°´æ±½æ¥æºç‰¹å¾.

We determined the quantity and source of soil water condensation versus atmospheric vapor condensation in Guanzhong Plain and explored their role in water balance, using three different types of hand-made micro-lysimeters (open-ends, top-seal, and bottom-seal designs). Field monitoring of the vapor condensation process was carried out from late September to late October in 2018 and from March to May in 2019 using the weighing method. Results showed that during the monitoring period, condensation occurred every day without rainfall event. The maximum daily condensation for the open-ends, top-seal and bottom-seal designs were 0.38, 0.27 and 0.16 mm, respectively, indicating that vapor flow in soil pores was the main source of soil water condensation, and that the measurement using the open-ended micro-lysimeter could faithfully reflect the amount of condensation in Guanzhong Plain. During the monitoring period, total soil water condensation reached 14.94 mm, accounting for 12.8% of the precipitation in the same period (116.4 mm), and the ratio of atmospheric vapor condensation to soil vapor condensation was 0.59:1.

Arula-7 powder improves diarrhea and intestinal epithelial tight junction function associated with its regulation of intestinal flora in calves infected with pathogenic Escherichia coli O1.

BACKGROUND: The effects of Arula-7 powder (ASP) on diarrhea and intestinal barrier function associated with its regulation of intestinal microflora in calves infected with pathogenic Escherichia coli O1 (E. coli O1) were studied. METHOD: Twenty Holstein calves were randomly divided into four treatment groups: normal control (NC), model control (MC), 0.5 mg/kg ciprofloxacin (CIP) and 2.50 g/kg ASP groups. RESULTS: ASP inhibited the relative abundance of Proteobacteria, Selenomonadales, and Enterobacteriales, and increased the relative abundance of Lactobacillus, Faecalibacterium, and Alloprevotella. Moreover, we demonstrated for the first time that the ASP and CIP promoted weight gain, reduced the diarrhea rate (P < 0.05), and enhanced antioxidant capacity (P < 0.05) due to the increase in average daily gain (ADG), total protein (TP), and albumin (ALB). In addition, ASP and CIP increased the expression of Zunola occludens-1 (ZO-1), Occludin, and Claudin-1 in the ileum (P < 0.05), and improved immunity due to increase levels of interleukin-2 (IL-2), interleukin-4 (IL-4), interferon-γ (IFN-γ), immunoglobulin A (IgA), and immunoglobulin G (IgG) in the serum, strengthened CD4+T levels in the ileal mucosa and reducing CD8+T and CD11c+T (P < 0.05). CONCLUSION: Hence, The intestinal microbiota environment formed by early intervention of ASP powder has a protective effect on the intestinal mucosal function of calves infected with pathogenic E. coli. Video Abstract.

Genome-wide identification and characterization of ABA receptor pyrabactin resistance 1-like protein (PYL) family in oat.

Abscisic acid (ABA) is a phytohormone that plays an important role in plant growth and development. Meanwhile, ABA also plays a key role in the plant response to abiotic stressors such as drought and high salinity. The pyrabactin resistance 1-like (PYR/PYL) protein family of ABA receptors is involved in the initial step of ABA signal transduction. However, no systematic studies of the PYL family in "Avena sativa, a genus Avena in the grass family Poaceae," have been conducted to date. Thus, in this study, we performed a genome-wide screening to identify PYL genes in oat and characterized their responses to drought stress. A total of 12 AsPYL genes distributed on nine chromosomes were identified. The phylogenetic analysis divided these AsPYLs into three subfamilies, based on structural and functional similarities. Gene and motif structure analysis of AsPYLs revealed that members of each subfamily share similar gene and motif structure. Segmental duplication appears to be the driving force for the expansion of PYLs, Furthermore, stress-responsive AsPYLs were detected through RNA-seq analysis. The qRT-PCR analysis of 10 AsPYL genes under drought, salt, and ABA stress revealed that AsPYL genes play an important role in stress response. These data provide a reference for further studies on the oat PYL gene family and its function.

Transcriptional Regulation of Different Rhizome Parts Reveal the Candidate Genes That Regulate Rhizome Development in Poa pratensis.

A strong rhizome can enhance the ability of a plant to resist drought, low temperature, and other stresses, as it can help plants rapidly obtain water and nutrients. Poa pratensis var. anceps Gaud. cv. Qinghai (QH) is a variant of P. pratensis that is widely distributed in natural grasslands above 3000 m above sea level on the Qinghai-Tibet Plateau. It forms turf easily and has strong soil-fixing ability due to its well-developed rhizomes. Understanding the molecular mechanism of rhizome development in this species is essential for cultivating new varieties of rhizome-type pasture for ecological protection. To clarify the transcriptional regulatory changes in different parts of the rhizome, we analyzed three different rhizome parts (rhizome buds, rhizome nodes, and rhizome internodes) of QH and weak-rhizome wild P. pratensis material (SN) using RNA sequencing. A total of 3806 genes were specifically expressed in Q_B, 1104 genes were specifically expressed in Q_N, and 1181 genes were specifically expressed in Q_I. Analysis showed that MYB, B3, NAC, BBR-BPC, AP2 MIKC_MADS, BSE1, and C2H2 may be key transcription factors regulating rhizome development. These genes interacted with multiple functional genes related to carbohydrate, secondary metabolism, and signal transduction, thus ensuring the normal development of the rhizomes. In particular, SUS (sucrose synthase) [EC:2.4.1.13] is specifically expressed in Q_I, which may be an inducing factor for the production of new plants from Q_B and Q_N. Additionally, PYL, PP2C, and SNRK2, which are involved in the abscisic acid signaling pathway, were differentially expressed in Q_N. In addition, genes related to protein modification and degradation, such as CIPKs, MAPKs, E2, and E3 ubiquitin ligases, were also involved in rhizome development. This study laid a foundation for further functional genomics studies on rhizome development in P. pratensis.

Freeze-thaw condition limits the fermentation process and accelerates the aerobic deterioration of oat (Avena sativa) silage in the Qinghai-Tibet Plateau.

The objective of this study is to determine the effect of freeze-thaw condition on the fermentation characteristics, microbial community, and aerobic stability of oat (Avena sativa) silage in the Qinghai-Tibet Plateau. Oat forage was harvested at milk ripening stage, ensiled in vacuum-sealed bags, and then stored at (1) a constant temperature of 20°C, as a control (20 group) or (2) subjected to freeze-thaw condition (alternating 20 and -5°C every 12 h; S group). The quality and microbial community in the silage were measured after 1, 3, 7, 14, and 60 days of ensiling, and the aerobic stability was measured after 60 days of ensiling at room temperature or at the two treatment temperatures. The results showed that the higher the pH, the lower the concentration of lactic acid and the ratio of lactic acid/acetic acid of the samples under freeze-thaw condition, as compared to those stored at 20°C. The dry matter content of 20 groups was significantly higher than S group (p < 0.05). While ash, neutral detergent fiber (NDF), acid detergent fiber (ADF), crude protein (CP), and water-soluble carbohydrates (WSC) had no significant difference between two groups. Lactobacillus spp., Leuconostoc spp., and Weissella spp. were the most prevalent bacterial genera in all groups. The abundance of Lactobacillus spp. in the 20 group was the highest on day 3 of ensiling (p < 0.05), and it reached the peak on day 14 in the S group, but the abundance in the S group did not exceed 50% during whole fermentation process. The abundance of Enterobacterales and the count of Escherichia coli in the S group was significantly higher than 20 group (p < 0.05). Interestingly, the lactic acid concentration was significant correlated with Lactobacillus spp. in 20 group, while correlated with Leuconostoc spp. in S group. The aerobic stability of the S group was lower than that of the 20 group (p < 0.05). The present study indicates that the freeze-thaw condition led to insufficient fermentation degree of silage by limiting the fermentation of Lactobacillus spp. and severely reduced the aerobic stability of oat silage.

Responses of photosynthetic characteristics of oat flag leaf and spike to drought stress.

Raising crops production via improving photosynthesis has always been focused. Recently excavating and increasing the photosynthetic capacity of non-leaf organs becomes an important approach to crops yield increase. Here we studied the photosynthetic characteristics of the flag leaf and the non-leaf organs including the sheath, the glume and the lemma under greenhouse. The relative water content (RWC), the stomatal characteristics, the photosynthetic pigment contents, the enzyme activities in C3 and C4 pathway and the malate content of the flag leaf and the non-leaf organs on 7, 14, 21, and 28 days after anthesis (denoted by 7DAA, 14DAA, 21DAA, and 28DAA) were determined under well-watered (CK) and water-stressed (D) treatments. Drought stress significantly reduced the RWC of the flag leaf and the non-leaf organs, while the variation of RWC in the glume and the lemma was lower than in the flag leaf. The chlorophyll a content, the chlorophyll b content, the total chlorophyll content and the xanthophyll content in the flag leaf were significantly decreased under D. However, drought stress significantly increased the photosynthetic pigment contents in the glume at the late stage (21DAA and 28DAA). In addition, the induced activities of PEPC, NADP-MDH, NADP-ME, NAD-ME, and PPDK in non-leaf organs under drought stress suggested that the C4 photosynthetic pathway in non-leaf organs compensated the limited C3 photosynthesis in the flag leaf. Non-leaf organs, in particular the glume, showed the crucial function in maintaining the stable photosynthetic performance of oat.

Corrigendum: Freeze-thaw condition limits the fermentation process and accelerates the aerobic deterioration of oat (Avena sativa) silage in the Qinghai-Tibet Plateau.

[This corrects the article DOI: 10.3389/fmicb.2022.944945.].

Genome-Wide Identification and Characterization of the Oat (Avena sativa L.) WRKY Transcription Factor Family.

The WRKY family is widely involved in the regulation of plant growth and stress response and is one of the largest gene families related to plant environmental adaptation. However, no systematic studies on the WRKY family in oat (Avena sativa L.) have been conducted to date. The recently published complete genome sequence of oat enables the systematic analysis of the AsWRKYs. Based on a genome-wide study of oat, we identified 162 AsWRKYs that were unevenly distributed across 21 chromosomes; a phylogenetic tree of WRKY domains divided these genes into three groups (I, II, and III). We also analyzed the gene duplication events and identified a total of 111 gene pairs that showed strong purifying selection during the evolutionary process. Surprisingly, almost all genes evolved after the completion of subgenomic differentiation of hexaploid oat. Further studies on the functional analysis indicated that AsWRKYs were widely involved in various biological processes. Notably, expression patterns of 16 AsWRKY genes revealed that the response of AsWRKYs were affected by stress level and time. In conclusion, this study provides a reference for further analysis of the role of WRKY transcription factors in species evolution and functional differentiation.

The effects of native lactic acid bacteria on the microbiome, fermentation profile, and nutritive value of Napier grass silage prepared with different legume ratios.

Mixing grass with legumes before ensiling is beneficial for improving dry matter and crude protein yield, but additional information is needed to balance nutrient content and fermentation quality. In this study, the microbial community, fermentation characteristics, and nutrient content of Napier grass mixed with alfalfa at different proportions were assessed. Tested proportions included: 100:0 (M0), 70:30 (M3), 50:50 (M5), 30:70 (M7), and 0:100 (MF). Treatments included: (CK) sterilized deionized water; (IN) selected lactic acid bacteria: Lactobacillus plantarum CGMCC 23166 and Lacticaseibacillus rhamnosus CGMCC 18233 (1.5 × 105 cfu/g of fresh weight for each inoculant); and (CO) commercial lactic acid bacteria: L. plantarum (1 × 105 cfu/g of fresh weight). All mixtures were ensiled for 60 days. Data analysis was used as a completely randomized design with a 5-by-3 factorial arrangement of treatments. The results showed that with increasing alfalfa mixing ratio, dry matter, and crude protein increased, while neutral detergent fiber and acid detergent fiber decreased both before and after ensiling (p < 0.05), which was not influenced by fermentation. Inoculation with IN and CO decreased pH and increased the lactic acid content compared to CK (p < 0.05), especially in silages M7 and MF. The highest Shannon index (6.24) and Simpson index (0.93) were observed in the MF silage CK treatment (p < 0.05). The relative abundance of Lactiplantibacillus decreased with increasing alfalfa mixing ratio, while the abundance of Lactiplantibacillus was significantly higher in the IN-treated group than in other treatment groups (p < 0.05). A higher alfalfa mixing ratio improved the nutrient value, but also made fermentation more difficult. Inoculants improved the fermentation quality by increasing the abundance of Lactiplantibacillus. In conclusion, the groups M3 and M5 achieved the optimal balance of nutrients and fermentation. If a higher proportion of alfalfa needs to be used, it is recommended to use inoculants to ensure sufficient fermentation.

Circular RNA circ_0079593 enhances malignant melanoma progression by the regulation of the miR-573/ABHD2 axis.

BACKGROUND: Malignant melanoma is the most fatal type of skin tumor. Circular RNAs (circRNAs) have been implicated in the malignant progression of melanoma. OBJECTIVE: The main purpose of this paper was to identify the precise parts of circ_0079593 in the malignant progression of melanoma. METHODS: The levels of circ_0079593, miR-573 and abhydrolase domain containing 2 (ABHD2) were determined by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Cell proliferation, colony formation, cell cycle progression, apoptosis, migration, and invasion were evaluated using the Cell Counting Kit-8 (CCK-8), colony formation, flow cytometry, and transwell assays, respectively. Targeted correlations among circ_0079593, miR-573 and ABHD2 were confirmed by dual-luciferase reporter, RNA immunoprecipitation (RIP) and RNA pull-down assays. Animal studies were performed to assess the role of circ_0079593 in vivo. RESULTS: Our data showed that circ_0079593 level was up-regulated in melanoma tissues and cells. The knockdown of circ_0079593 suppressed cell proliferation, cell cycle progression, migration, invasion, and enhanced apoptosis in vitro and inhibited tumor growth in vivo. Mechanistically, circ_0079593 directly targeted miR-573, and circ_0079593 controlled ABHD2 expression by miR-573. MiR-573 mediated the regulation of circ_0079593 on melanoma cell progression in vitro. Moreover, ABHD2 was a functional target of miR-573 in regulating melanoma cell progression in vitro. CONCLUSION: Our findings identified that the knockdown of circ_0079593 suppressed melanoma progression at least partially through targeting the miR-573/ABHD2 axis, providing evidence for developing circ_0079593 as a promising therapeutic target for melanoma treatment.