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Abstract Background Soybean (Glycine max L.) is one of the most important oil crops in the world. It is desirable to increase oil yields from soybean, and so this has been a major goal of oilseed engineering. However, it is still uncertain how many genes and which genes are involved in lipid biosynthesis. Results Here, we evaluated changes in gene expression over the course of seed development using Illumina (formerly Solexa) RNA-sequencing. Tissues at 15 days after flowering (DAF) served as the control, and a total of 11592, 16594, and 16255 differentially expressed unigenes were identified at 35, 55, and 65 DAF, respectively. Gene Ontology analyses detected 113 co-expressed unigenes associated with lipid biosynthesis. Of these, 15 showed significant changes in expression levels (log2fold values ≥ 1) during seed development. Pathway analysis revealed 24 co-expressed transcripts involved in lipid biosynthesis and fatty acid biosynthesis pathways. We selected 12 differentially expressed genes and analyzed their expressions using qRT-PCR. The results were consistent with those obtained from Solexa sequencing. Conclusion These results provide a comprehensive molecular biology background for research on soybean seed development, particularly with respect to the process of oil accumulation. All of the genes identified in our research have significance for breeding soybeans with increased oil contents.

Background: Bone loss caused by microgravity exposure presents a serious threat to the health of astronauts, but existing treatment strategies have specific restrictions. This research aimed to investigate whether salidroside (SAL) can mitigate microgravity-induced bone loss and its underlying mechanism. Methods: In this research, we used hindlimb unloading (HLU) and the Rotary Cell Culture System (RCCS) to imitate microgravity in vivo and in vitro. Results: The results showed that salidroside primarily enhances bone density, microstructure, and biomechanical properties by stimulating bone formation and suppressing bone resorption, thereby preserving bone mass in HLU rats. In MC3T3-E1 cells cultured under simulated microgravity in rotary wall vessel bioreactors, the expression of osteogenic genes significantly increased after salidroside administration, indicating that salidroside can promote osteoblast differentiation under microgravity conditions. Furthermore, the Nrf2 inhibitor ML385 diminished the therapeutic impact of salidroside on microgravity-induced bone loss. Overall, this research provides the first evidence that salidroside can mitigate bone loss induced by microgravity exposure through stimulating the Nrf2/HO-1 pathway. Conclusion: These findings indicate that salidroside has great potential for treating space-related bone loss in astronauts and suggest that Nrf2/HO-1 is a viable target for counteracting microgravity-induced bone damage. [ABSTRACT FROM AUTHOR]

Long noncoding RNAs (LncRNAs) play essential roles in regulating gene expression in various biological processes. However, the function of lncRNAs in vascular smooth muscle cell (VSMC) transformation remains to be explained. In this work, we discover that a new bone marrow protein (BMP) signaling target, lncRNA RP11‐301G19.1, is significantly induced in BMP7‐treated VSMCs through lncRNA microarray analysis. Addition of BMP signaling inhibitor LDN‐193189 attenuates the expression of ACTA2 and SM‐22α, as well as the mRNA level of RP11‐301G19.1. Furthermore, lncRNA RP11‐301G19.1 is critical to the VSMC differentiation and is directly activated by SMAD1/9. Mechanistically, knocking down of RP11‐301G19.1 leads to the decrease of ATOH8, another BMP target, while the forced expression of RP11‐301G19.1 reactivates ATOH8. In addition, miR‐17‐5p, a miRNA negatively regulated by BMP‐7, contains predicted binding sites for lncRNA RP11‐301G19.1 and ATOH8 3′UTR. Accordingly, overexpression of miR‐17‐5p decreases the levels of them. Together, our results revealed the role of lncRNA RP11‐301G19.1 as a miRNA sponge to upregulate ATOH8 in VSMC phenotype transformation. [ABSTRACT FROM AUTHOR]

SUMMARY: Citrus is a model plant for studying adventitious embryos, a form of asexual reproduction controlled by a single dominant gene, RWP. This gene has been identified as the causal gene for nucellar embryogenesis, but its function has not yet been fully understood. In this study, we used the fast‐growing Fortunella hindsii as a system to explore chromatin accessibility during the nucellar embryony initiation, emphasizing elevated chromatin accessibility in polyembryonic (PO) genotypes compared to monoembryonic ones (MO). Notably, a higher level of accessible chromatin was observed in one allele of the promoter region of FhRWP, consistent with increased expression of the allele carrying the causal structural variant. By independently performing RNAi and gene editing experiments on PO genotypes, we found the downregulation of FhRWP expression could reduce the number of nucellar embryos, while its knockout resulted in abnormal axillary bud development. In overexpression experiments, FhRWP was identified as having the unique capability of inducing the embryogenic callus formation in MO stem segments, possibly through the regulation of the WUS‐CLV signaling network and the ABA and cytokinin pathway, marking the inaugural demonstration of FhRWP's potential to reignite somatic cells' embryogenic fate. This study reveals the pleiotropic function of RWP in citrus and constructs a regulatory network during adventitious embryo formation, providing a new tool for bioengineering applications in plant regeneration. Significance Statement: FhRWP plays a central role in regulating multiple developmental phenotypes, including axillary bud differentiation, nucellar embryo formation, and somatic embryogenesis in citrus. [ABSTRACT FROM AUTHOR]

Summary: MLL‐rearranged (MLL‐r) leukaemia is observed in approximately 10% of acute myeloid leukaemia (AML) and is associated with a relatively poor prognosis, highlighting the need for new treatment regimens. MLL fusion proteins produced by MLL rearrangements recruit KDM4C to mediate epigenetic reprogramming, which is required for the maintenance of MLL‐r leukaemia. In this study, we used a combinatorial drug screen to selectively identify synergistic treatment partners for the KDM4C inhibitor SD70. The results showed that the drug combination of SD70 and MI‐503, a potent menin‐MLL inhibitor, induced synergistically enhanced apoptosis in MLL::AF9 leukaemia cells without affecting normal CD34+ cells. In vivo treatment with SD70 and MI‐503 significantly prolonged survival in AML xenograft models. Differential gene expression analysis by RNA‐seq following combined pharmacological inhibition of SD70 and MI‐503 revealed changes in numerous genes, with MYC target genes being the most significantly downregulated. Taken together, these data provide preclinical evidence that the combination of SD70 and MI‐503 is a potential dual‐targeted therapy for MLL::AF9 AML. [ABSTRACT FROM AUTHOR]

Diseases like obesity and intestinal inflammation diseases are accompanied by dysbiosis of the gut microbiota (DSGM), which leads to various complications, including systemic metabolic disorders. DSGM reportedly impairs the fertility of male mice; however, the regulatory mechanism is unclear. Exosomes are molecular mediators of intercellular communication, but the regulation of spermatogenesis by non‐reproductive tissue‐originated exosomes remains unknown. The present study shows that DSGM altered the miRNA expression profile of mouse circulating exosomes and impaired spermatogenesis. Moreover, the single‐cell sequencing results indicate that circulating exosomes from mice with DSGM impaired spermatogenesis, while circulating exosomes from wild mice improved spermatogenesis by promoting meiosis. Further study demonstrates that DSGM leads to abnormal upregulation of miR‐211‐5p in gut‐derived circulating exosomes, which inhibited the expression of meiosis‐specific with coiled‐coil domain (Meioc) in the testes and impaired spermatogenesis by disturbing meiosis process. In summary, this study defines the important role of gut‐derived exosomes in connecting the "gut‐testis" axis. [ABSTRACT FROM AUTHOR]

• CircMYBL1 suppressed acquired resistance to osimertinib in non-small-cell lung cancer cells. • The results suggest that circMYBL1 could play an important role in facilitating growth and metastasis in susceptible cells. • In-silico analysis and the results show that the target genes of circMYBL1 are involved in tumor growth and metastasis. Circular RNAs (circRNAs) play an important role in the development of acquired resistance to many anticancer drugs. We developed the Non-Small-Cell Lung Cancer (NSCLC) cell lines with acquired resistance to osimertinib, a third-generation of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs), and evaluated the different expression profiles of circRNAs in osimertinib-sensitive and -resistant NSCLC cell lines using RNA sequencing (RNA-Seq). The expression of selected differentially expressed circRNAs was verified using quantitative real-time PCR (qRT-PCR) in paired osimertinib-sensitive and -resistant NSCLC cell lines, and in plasma samples of osimertinib-sensitive and -resistant NSCLC patients. We found that circMYBL1(has_circ_0136924) was downregulated after acquired resistance to osimertinib, inhibiting circMYBL1 expression facilitated the proliferation, migration, and invasion in osimertinib-sensitive NSCLC cells. CircMYBL1 may be a novel molecular biomarker and therapeutic target for osimertinib-resistant NSCLC. [ABSTRACT FROM AUTHOR]

Acetohydroxyacid synthase (AHAS) is a key enzyme in the first step of the branched-chain amino acid synthesis pathway, and the production of acetohydroxybutyrate from one molecule of 2-ketobutyric acid and one molecule of pyruvate. AHAS is inhibited by feedback from L-valine, L-leucine, and L-isoleucine, and the expression of ilvBN, the gene encoding AHAS, is regulated by all three branched-chain amino acids. A change in amino acids 20–22 on the regulatory subunit (M13 mutation) removes the feedback inhibition by valine. We cloned the gene encoding AHAS (ilvBN) into a vector and then transfected it into Escherichia coli BL21 for expression with targeted changes in amino acids 20–22 on the regulatory subunit, and then determined the activity of the mutated AHAS and its inhibitory effects on valine, isoleucine, and leucine. The enzyme containing the M13 mutation was feedback resistant to all three amino acids. Previous studies have suggested that the binding sites for the three branched-chain amino acids may be at the same variable center. We investigated the enzymatic properties of wild-type and mutant AHAS, modeled their crystal structures, and resolved the mechanism of feedback inhibition induced by mutant M13, which will be useful for continuing the modification of AHAS and the design of broad-spectrum herbicides. [ABSTRACT FROM AUTHOR]

Simple Summary: MicroRNAs (miRNAs) play important roles in the regulation of a plant's response to a variety of pathogens. However, the regulatory networks of miRNAs and their targets in pepper's response to P. capsici are largely unknown. Herein, integrative miRNA-seq and mRNA-seq analysis were performed to identify miRNAs and their target genes in pepper during P. capsici infection, and the results provide a valuable resource for better understanding the regulatory mechanisms between miRNAs and their targets in pepper's defense against P. capsici. Phytophthora blight of pepper is a notorious disease caused by the oomycete pathogen Phytophthora capsici, which poses a great threat to global pepper production. MicroRNA (miRNA) is a class of non-coding small RNAs that regulate gene expressions by altering the translation efficiency or stability of targeted mRNAs, which play important roles in the regulation of a plant's response to pathogens. Herein, time-series mRNA-seq libraries and small RNA-seq libraries were constructed using pepper roots from the resistant line CM334 and the susceptible line EC01 inoculated with P. capsici at 0, 6, 24, and 48 h post-inoculation, respectively. For mRNA-seq analysis, a total of 2159 and 2971 differentially expressed genes (DEGs) were identified in CM334 and EC01, respectively. For miRNA-seq analysis, 491 pepper miRNAs were identified, including 330 known miRNAs and 161 novel miRNAs. Among them, 69 and 88 differentially expressed miRNAs (DEMs) were identified in CM334 and EC01, respectively. Examination of DEMs and their targets revealed 22 regulatory networks, predominantly featuring up-regulated miRNAs corresponding to down-regulated target genes. Notably, these DEM-DEG regulatory networks exhibited significant overlap between CM334 and EC01, suggesting that they might contribute to pepper's basal defense against P. capsici. Furthermore, five selected DEMs (miR166, miR1171, miR395, miR530 and miRN2) and their target genes underwent qRT-PCR validation, confirming a consistent negative correlation in the expression patterns of miRNAs and their targets. This comprehensive analysis provides novel insights into the regulatory networks of miRNAs and their targets, offering valuable contributions to our understanding of pepper's defense mechanisms against P. capsici. [ABSTRACT FROM AUTHOR]

The disorders of endometrial receptivity and ovulatory dysfunction are both significant causes of infertility in patients with polycystic ovary syndrome (PCOS). In this study, we investigated the expression profile and functional implications of circular RNAs (circRNAs) in the endometrial receptivity of PCOS‐affected mice. Twenty‐four female C57BL/6 mice were divided into PCOS and normal control groups. The PCOS group received subcutaneous DHEA treatment, while the control group remained untreated. Gene chip technology was utilized to analyze circRNA expression in endometrial tissues on the fourth day of gestation with subsequent bioinformatics analyses into circRNA functions. Furthermore, endometrial epithelial cells were used to determine represented circRNA functions. Results showed that the PCOS group exhibited 205 differentially expressed circRNAs, with 147 upregulated and 58 downregulated ones. qRT‐PCR confirmed differential expression of circRNAs, including circRNA_38548, circRNA_001686, circRNA_38550, and circRNA_27938. Predicted target genes and a circRNA‐miRNA‐mRNA regulatory network were constructed. Additionally, four circRNAs (circRNA_38548, circRNA_38550, and circRNA_001686) were identified to contribute to abnormal endometrial receptivity by regulating genes such as Lifr, FOXK1, FOXO1, HOXA10, through interactions with miRNAs. Further research is warranted to elucidate the underlying mechanisms involving these circRNAs. [ABSTRACT FROM AUTHOR]

During fertilization, DAXX (death domain‐associated protein) mediates histone variant H3.3 incorporation into heterochromatin, which plays an important role in the maintenance of genomic integrity. rDNA, the ribosomal gene, is included in the first wave of gene activation after fertilization. Our and other studies indicated that loss of Daxx disturbs rDNA heterochromatinization and promotes rDNA transcription without change in protein expression of H3.3. However, maternal and zygotic deletion of Daxx impairs blastocyst development. Whether Daxx knockdown affects H3.3 expression and improves the rDNA transcription in preimplantation development has not been reported. In the present study, we injected HA‐labelled H3.3 (H3.3‐HA) into oocytes during ICSI procedure, and detected H3.3 and DAXX by immunofluorescent staining. Then, we knockdowned Daxx and detected the gene expression levels of Daxx, H3.3, 18s and 47s rRNA. We also performed immunofluorescent staining of B23, γH2A and EdU incorporation to demonstrate nuclear structure, DNA damage and replication. We found injection of H3.3‐HA did not impair preimplantation development. Daxx siRNA did not change expression of H3.3 mRNA, and the development of two‐cell embryos and blastocysts, but the overall replication and expression levels of rRNA were increased compared with that in the control group. Finally, knockdown of DAXX did not aggravate the DNA damage but loosened the nucleolus. We concluded that Daxx knockdown promoted DNA replication and rDNA transcription, but did not affect H3.3 expression and subsequent preimplantation development. [ABSTRACT FROM AUTHOR]

Flowers and pod abscission significantly reduces soybean yield. This study aims to identify the main signaling pathways and key candidate genes in soybean leaves that affect flower and pod abscission under shade conditions. This information will be useful for the localization and cloning of genes related to abscission. Two soybean cultivars with different abscission rates (Liaodou 32 and Shennongdou 28) were used in this experiment. The soybean leaves were subjected to 50% shading treatment and the transcriptome and metabolome was sequenced during the light-sensitive period. The effects of weak light at different growth stages on the metabolic pathways of soybean leaves and organ abscission were investigated by analyzing plant phenotype and physiological changes. The results showed that both two cultivars triggered the same molecular mechanism and similar metabolite accumulation mode by shading, but the regulations of the two cultivars were different. The key candidate genes identified for soybean flower and pod abscission caused by shading were DIV, MYB06, MYB44, MY1R1, MYBS3, WRKY6, WRK53, WRK70, WRK40, DOF14, CDF3, CDF2, GATA5, DREB3 and ERF3; the differentially expressed genes that caused the variation between the cultivars were SRM1, MYB16, WRK24, COL16, MYB61 and TRB1. The main metabolic pathways involved in soybean flower and pod abscission caused by shading were secondary metabolite biosynthesis pathway, metabolic pathway, cofactor biosynthesis pathway, phenylpropanoid biosynthesis pathway, flavonoid biosynthesis pathway, fatty acid biosynthesis pathway and amino acid biosynthesis pathway; the DEMs that caused the differences among the cultivars were carbon metabolism, glutathione metabolism, biotin metabolism, nucleotide metabolism, purine metabolism, plant hormone signaling, flavone and flavonol biosynthesis, lysine degradation, arginine and proline metabolism, amino sugars and nucleotide sugars metabolism, etc. In conclusion, shading treatment in the light-sensitive period of soybean changed the physiological response and gene expression level of leaves, inhibited carbohydrate metabolism and transport and biosynthesis of secondary metabolites, and thereby leading to increased competition and hormonal disruption, which promoted the abscission of soybean flowers/pods and reduced grain yield. [ABSTRACT FROM AUTHOR]

The paper mulberry is a commonly found tree species with a long history of cultivation. It also serves as a crucial case study for understanding how woody plants adapt to low temperatures. Under cold treatment, we observed a substantial number of alternative splicing (AS) genes, showcasing the intricate landscape of AS events. We have detected all seven types of AS events, with the alternative 3′ splice site (A3) having the most. We observed that many genes that underwent differential AS were significantly enriched in starch and sucrose metabolism and circadian rhythm pathways. Moreover, a considerable proportion of differentially spliced genes (DSGs) also showed differential expression, with 20.38% and 25.65% under 12 h and 24 h cold treatments, respectively. This suggests a coordinated regulation between gene AS and expression, playing a pivotal role in the paper mulberry's adaptation to cold stress. We further investigated the regulatory mechanisms of AS, identifying 41 serine/arginine-rich (SR) splicing factors, among which 11 showed differential expression under cold treatment, while 29 underwent alternative splicing. Additionally, genes undergoing AS displayed significantly higher DNA methylation levels under cold stress, while normal splicing (non-AS) genes exhibited relatively lower methylation levels. These findings suggest that methylation may play an important role in governing gene AS. Finally, our research will provide useful information on the role of AS in the cold acclimation tolerance of the paper mulberry. [ABSTRACT FROM AUTHOR]

Species residing across elevational gradients display adaptations in response to environmental changes such as oxygen availability, ultraviolet radiation, and temperature. Here, we study genomic variation, gene expression, and long-term adaptation in Tibetan Partridge (Perdix hodgsoniae) populations residing across the elevational gradient of the Tibetan Plateau. We generated a high-quality draft genome and used it to carry out downstream population genomic and transcriptomic analysis. The P. hodgsoniae populations residing across various elevations were genetically distinct, and their phylogenetic clustering was consistent with their geographic distribution. We identified possible evidence of gene flow between populations residing in <3,000 and >4,200 m elevation that is consistent with known habitat expansion of high-altitude populations of P. hodgsoniae to a lower elevation. We identified a 60 kb haplotype encompassing the Estrogen Receptor 1 (ESR1) gene, showing strong genetic divergence between populations of P. hodgsoniae. We identified six single nucleotide polymorphisms within the ESR1 gene fixed for derived alleles in high-altitude populations that are strongly conserved across vertebrates. We also compared blood transcriptome profiles and identified differentially expressed genes (such as GAPDH , LDHA , and ALDOC) that correlated with differences in altitude among populations of P. hodgsoniae. These candidate genes from population genomics and transcriptomics analysis were enriched for neutrophil degranulation and glycolysis pathways, which are known to respond to hypoxia and hence may contribute to long-term adaptation to high altitudes in P. hodgsoniae. Our results highlight Tibetan Partridges as a useful model to study molecular mechanisms underlying long-term adaptation to high altitudes. [ABSTRACT FROM AUTHOR]

Northern corn leaf blight, caused by the fungal pathogen Exserohilum turcicum, is a major disease of maize. The first major locus conferring resistance to E. turcicum race 0, Ht1, was identified over 50 years ago, but the underlying gene has remained unknown. We employed a map‐based cloning strategy to identify the Ht1 causal gene, which was found to be a coiled‐coil nucleotide‐binding, leucine‐rich repeat (NLR) gene, which we named PH4GP‐Ht1. Transgenic testing confirmed that introducing the native PH4GP‐Ht1 sequence to a susceptible maize variety resulted in resistance to E. turcicum race 0. A survey of the maize nested association mapping genomes revealed that susceptible Ht1 alleles had very low to no expression of the gene. Overexpression of the susceptible B73 allele, however, did not result in resistant plants, indicating that sequence variations may underlie the difference between resistant and susceptible phenotypes. Modelling of the PH4GP‐Ht1 protein indicated that it has structural homology to the Arabidopsis NLR resistance gene ZAR1, and probably forms a similar homopentamer structure following activation. RNA sequencing data from an infection time course revealed that 1 week after inoculation there was a threefold reduction in fungal biomass in the PH4GP‐Ht1 transgenic plants compared to wild‐type plants. Furthermore, PH4GP‐Ht1 transgenics had significantly more inoculation‐responsive differentially expressed genes than wild‐type plants, with enrichment seen in genes associated with both defence and photosynthesis. These results demonstrate that the NLR PH4GP‐Ht1 is the causal gene underlying Ht1, which represents a different mode of action compared to the previously reported wall‐associated kinase northern corn leaf blight resistance gene Htn1/Ht2/Ht3. [ABSTRACT FROM AUTHOR]

Leaf margin serration is a morphological characteristic in plants. The CUC2 (CUP-SHAPED COTYLEDON 2) gene plays an important role in the outgrowth of leaf teeth and enhances leaf serration via suppression of growth in the sinus. In this study, we isolated the BcCUC2 gene from Pak-choi (Brassica rapa ssp. chinensis), which contains a 1104 bp coding sequence, encoding 367 amino acid residues. Multiple sequence alignment exhibited that the BcCUC2 gene has a typical conserved NAC domain, and phylogenetic relationship analysis showed that the BcCUC2 protein has high identity with Cruciferae plants (Brassica oleracea, Arabidopsis thaliana, and Cardamine hirsuta). The tissue-specific expression analysis displayed that the BcCUC2 gene has relatively high transcript abundance in floral organs. Meanwhile, the expression profile of BcCUC2 was relatively higher in the '082' lines with serrate leaf margins than the '001' lines with smooth leaf margins in young leaves, roots, and hypocotyls. In addition, the transcript level of BcCUC2 was up-regulated by IAA and GA3 treatment, especially at 1–3 h. The subcellular localization assay demonstrated that BcCUC2 was a nuclear-target protein. Furthermore, leaf serration occurred, and the number of the inflorescence stem was increased in the transgenic Arabidopsis thaliana plants' overexpressed BcCUC2 gene. These data illustrated that BcCUC2 is involved in the development of leaf margin serration, lateral branches, and floral organs, contributing to further uncovering and perfecting the regulation mechanism of leaf serration in Pak-choi. [ABSTRACT FROM AUTHOR]

Flavonoids are the most abundant class of secondary metabolites that are ubiquitously involved in plant development and resistance to biotic and abiotic stresses. Flavonoid biosynthesis involves multiple channels of orchestrated molecular regulatory factors. Methyl jasmonate (MeJA) has been demonstrated to enhance flavonoid accumulation in numerous plant species; however, the underlying molecular mechanism of MeJA-induced flavonoid biosynthesis in safflower is still not evident. In the present study, we revealed the underlying molecular basis of a putative F3′5′H gene from safflower imparting MeJA-induced flavonoid accumulation in transgenic plants. The constitutive expression of the CtF3′5′H1 gene was validated at different flowering stages, indicating their diverse transcriptional regulation through flower development in safflower. Similarly, the CtF3′5′H1-overexpressed Arabidopsis plants exhibit a higher expression level, with significantly increased anthocyanins and flavonoid content, but less proanthocyanidins than wild-type plants. In addition, transgenic plants treated with exogenous MeJA revealed the up-regulation of CtF3′5′H1 expression over different time points with significantly enhanced anthocyanin and flavonoid content as confirmed by HPLC analysis. Moreover, CtF3′5′H1- overexpressed Arabidopsis plants under methyl violet and UV-B irradiation also indicated significant increase in the expression level of CtF3′5′H1 with improved anthocyanin and flavonoid content, respectively. Noticeably, the virus-induced gene silencing (VIGS) assay of CtF3′5′H1 in safflower leaves also confirmed reduced anthocyanin accumulation. However, the CtF3′5′H1 suppression in safflower leaves under MeJA elicitation demonstrated significant increase in total flavonoid content. Together, our findings confirmed that CtF3′5′H1 is likely mediating methyl jasmonate-induced flavonoid biosynthesis in transgenic plants via enhanced anthocyanin accumulation. [ABSTRACT FROM AUTHOR]

'Shannongsu' and 'Xincixiang' were two new late-ripening pear cultivars with high quality, that were bred by our team. In order to clear the postharvest storage characteristics, mature pears were collected and stored at room temperature. Several fruit characteristics were evaluated over time, such as firmness, ethylene release rate, content of aroma components, softening-related enzyme activity, and gene expression. Both 'Shannongsu' and 'Xincixiang' were crisp and juicy stored after 60 d at room temperature, which exhibited excellent storage characteristics. The storability may be comprehensive result of low levels of ethylene production, aroma synthesis, softening-related activities, and gene expression. The research cleared the storage characteristics of 'Shannongsu' and 'Xincixiang' at room temperature for the first time, which will provide scientific theoretical guidance for fruit production and marketing. [ABSTRACT FROM AUTHOR]

WRKY transcription factors (TF) control a variety of biological processes in higher plants, including abiotic and biotic stress responses. WRKY genes have been identified and functionally characterized in many plant species during the past ten years, but despite this progress, few investigations have focused on this TF family in licorice (Glycyrrhiza uralensis Fisch.). This study identified 72 WRKY genes from the licorice genome and classified them into three groups. The gene structure analysis indicated that the GuWRKY genes contain from one to eight introns, with the majority having two introns and three exons. Conserved motif analysis identified the conserved WRKY domain in most GuWRKYs. Further analysis of the promoter region revealed that almost all GuWRKYs had plant hormone- or stress-responsive cis-regulatory elements, with differences in the number of elements among the groups. The role of 10 selected GuWRKYs in regulating abiotic stress responses was further explored via quantitative real-time PCR (qRT-PCR) after low-temperature (4 °C), salt (150 mmol·L−1 NaCl), drought (15% polyethylene glycol 6000) treatments at 3, 6, 12, and 24 h. Thus, our study of the WRKY gene family in licorice provides a basis for further research on the structure and function of WRKY genes. [ABSTRACT FROM AUTHOR]

Pseudogene-derived transcripts, especially those barely transcribed in normal tissues, have been regarded as a kind of non-coding RNAs, and present potential functions in tumorigenicity and tumor development in human beings. However, their exact effects on hepatocellular carcinoma (HCC) remain largely unknown. On basis of our previous research and the constructed online database for the non-coding RNAs related to HCC, a series of pseudogene transcripts have been discovered, and SNRPFP1, the homologous pseudogene of SNRPF, was found to produce an anomalously high expression long non-coding RNA in HCC. In this study, we validated the expression of the SNRPFP1 transcript in both HCC tissues and cell lines. The adverse correlation between SNRPFP1 expression and patients' outcomes was observed. And depletion of SNRPF1 in HCC cells significantly suppressed cell proliferation and apoptosis resistance. Meanwhile, the motility of HCC cells was potently impaired. Interestingly, miR-126-5p, one of the tumor-suppressive genes commonly decreased in HCC, was found negatively expressed and correlated with SNRPF1, and a specific region of SNRPF1 transcript is directly binding to miR-126-5p in a molecular sponge way. The rescue experiment by knock-out miR-126-5p significantly reversed the cell growth suppression and a higher ratio of cell apoptosis induced by SNRPF1 depletion. Lastly, we concluded that SNRPF1 is a pseudogene active in HCC, and its abnormally over-expressed transcript is a strong promoter of HCC cell progress in vitro by sponging miR-126-5p. We believe that the findings in this study provide new strategies for HCC prevention and therapeutic treatment. [ABSTRACT FROM AUTHOR]

Ulcerative colitis (UC), a chronic inflammatory disease, often cause carcinogenesis, disability, and intestinal perforation. The JingFangFuZiLiZhong formula (JFFZLZ) shows a good effect against UC in the clinic. Hence, we aim to investigate the mechanisms between JFFZLZ and UC via network pharmacology data mining and in vivo experiments. We obtained active constituents and related targets from public databases. The overlapped genes between JFFZLZ and UC targets were further analysed by enrichment analysis. The active constituents and hub targets were used to construct molecule docking analysis. We finally screened out nine hub targets and their expressions were verified in the Gene Expression Omnibus database and UC rats' colon tissues after JFFZLZ treatment. The results implied that JFFZLZ mainly regulated signal transduction, metabolites production, and inflammation pathways. The expression of STAT3, CXCL8, IL6, CXCL12, TNF, TP53, and PTPN11 were both upregulated in colon tissues of UC patients and UC rats. While RELA, EGFR, and TP53 were downregulated in UC patients, but upregulated in UC rats. Furthermore, JFFZLZ could repair UC rats' colon mucosal damage and promote the healing of ulcers via regulating the hub targets. These results elucidated that the anti-UC effect of JFFZLZ was closely related to the inhibition of inflammatory response, inhibition of oxidative stress, and repairing colon mucosal damage through different signal pathways. The findings could contribute to a better understanding of the regulation mechanisms in JFFZLZ against UC. JFFZLZ could reduce the inflammatory infiltration and repair UC rats' colon mucosal damage. Through the network pharmacology-based strategy and public database mining, we obtained the hub targets and key pathways between JFFZLF and UC. The mechanism of JFFZLZ against UC was inhibition of inflammatory response and oxidative stress by regulating the expression of the hub targets. [ABSTRACT FROM AUTHOR]

Background. Ferroptosis, a type of cell death caused by phospholipid peroxidation, has lately been linked to the onset and development of numerous illnesses. Numerous investigations have demonstrated the close relationship between lipid peroxidation and carotid atherosclerosis. In order to get new knowledge for targeted therapy, bioinformatics analysis was employed in this study to discover the probable ferroptosis-related genes of carotid atherosclerosis. Methods. The GSE43292 gene expression dataset was downloaded from the Gene Expression Omnibus (GEO) database. The differentially expressed ferroptosis-related genes were screened by R software and then analyzed by protein-protein interaction (PPI) network, differential gene correlation analysis, Kyoto Encyclopedia of Gene and Genome (KEGG) pathway, and Gene Ontology (GO) terminology enrichment analysis to explore the functional role. Result. In samples of atherosclerosis, we found 33 ferroptosis genes that were differentially expressed, including 21 upregulated genes and 12 downregulated genes. These differentially elevated genes were mainly connected to the ferroptosis and glutathione metabolism pathways, according to GO and KEGG enrichment analysis. We also discovered 10 hub genes and 2 important modules through the analysis of the PPI network and the creation of key modules. Conclusion. The current findings imply that the carotid atherosclerosis phenomenon involves ferroptosis, and 10 important genes associated to ferroptosis may play a role in the development of carotid atherosclerosis. This study offered a novel approach to future research on the carotid atherosclerosis pathogenic processes and treatment targets. [ABSTRACT FROM AUTHOR]

• The lifestyle for brain health (LIBRA) index has been associated with cognitive function. • Whether CD33 rs3865444 polymorphism can modulate the association between the LIBRA index and cognition in dementia-free older adults is unknown. • Our study suggested that a higher LIBRA index was associated with worse cognitive performance in rural Chinese older adults, especially among the CD33 CC carriers. • This study contributes to our further understanding of the complex relationships of modifiable lifestyles and the CD33 gene with cognitive function, which has potential implications for preventive interventions to maintain late-life brain and cognitive health. We sought to examine the associations of the Lifestyle for Brain Health (LIBRA) index with cognitive function among rural Chinese older adults and to explore the potential role of cluster of differentiation 33 gene (CD33) in the associations. This population-based cross-sectional study included 4914 dementia-free participants (age ≥60 years; 56.43 % women) in the 2018 baseline examination of MIND-China. The LIBRA index was generated from 11 factors. We used a neuropsychological test battery to assess episodic memory, verbal fluency, attention, executive function, and global cognition. The CD33 (rs3865444) polymorphism was detected using multiple-polymerase chain reaction amplification. Data were analyzed using the general linear regression models. A higher LIBRA index was associated with multivariable-adjusted β-coefficient (95 %CI) of -0.011(-0.020- -0.001) for global cognitive z-score, -0.020(-0.033- -0.006) for episodic memory, and -0.016(-0.029- -0.004) for verbal fluency. The CD33 (rs3865444) was associated with a lower global cognitive z-score in the additive (CA vs. CC: β-coefficient=0.042; 95 %CI=0.008–0.077), the dominant (CA+AA vs. CC: 0.040; 0.007–0.073), and the over-dominant (CA vs. CC+AA: 0.043; 0.009–0.077) models. Similar results were obtained for verbal fluency and attention. The CD33 gene showed statistical interactions with LIBRA index on cognitive function (P interaction <0.05) such that a higher LIBRA index was significantly associated with lower z-scores of global cognition and attention only among CD33 CC carriers (P < 0.05). This population-based study reveals for the first time that a higher LIBRA index is associated with worse cognitive performance in rural Chinese older adults and that CD33 gene could modify the association. [ABSTRACT FROM AUTHOR]

Background: Breast cancer (BC) is the most common malignant tumor worldwide. Apoptosis and hypoxia are involved in the progression of BC, but reliable biomarkers for these have not been developed. We hope to explore a gene signature that combined apoptosis and hypoxia‐related genes (AHGs) to predict BC prognosis and immune infiltration. Methods: We collected the mRNA expression profiles and clinical data information of BC patients from The Cancer Genome Atlas database. The gene signature based on AHGs was constructed using the univariate Cox regression, least absolute shrinkage and selection operator, and multivariate Cox regression analysis. The associations between risk scores, immune infiltration, and immune checkpoint gene expression were studied using single‐sample gene set enrichment analysis. Besides, gene signature and independent clinicopathological characteristics were combined to establish a nomogram. Finally, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were performed on the potential functions of AHGs. Results: We identified a 16‐AHG signature (AGPAT1, BTBD6, EIF4EBP1, ERRFI1, FAM114A1, GRIP1, IRF2, JAK1, MAP2K6, MCTS1, NFKBIA, NFKBIZ, NUP43, PGK1, RCL1, and SGCE) that could independently predict BC prognosis. The median score of the risk model divided the patients into two subgroups. By contrast, patients in the high‐risk group had poorer prognosis, less abundance of immune cell infiltration, and expression of immune checkpoint genes. The gene signature and nomogram had good predictive effects on the overall survival of BC patients. GO and KEGG analyses revealed that the differential expression of AHGs may be closely related to tumor immunity. Conclusion: We established and verified a 16‐AHG BC signature which may help predict prognosis, assess potential immunotherapy benefits, and provide inspiration for future research on the functions and mechanisms of AHGs in BC. [ABSTRACT FROM AUTHOR]

Background: Intervertebral disc degeneration (IDD) has become a serious public health problem, the mechanism of which is complex and still unclear. We aimed to construct a ceRNA network related to IDD to explore its pathogenesis. Methods: We downloaded the GSE67566, GSE63492, GSE116726 and GSE124272 datasets from GEO database, and obtained the differentially expressed RNAs. Then, we constructed a ceRNA network and the KEGG and GO enrichment analysis were performed. Finally, we performed immune cell infiltration analysis on the GSE124272 dataset and analysed the correlation between immune cell abundance and hub genes expression levels. Results: The ceRNA network included three down-regulated circRNAs: hsa_circ_0074817, hsa_circ_0002702, hsa_circ_0003600, three up-regulated miRNAs: hsa-miR-4741, hsa-miR-3158-5p, hsa-miR-508-5p, and 57 down-regulated mRNAs, including six hub genes: IGF1, CHEK1, CCNB1, OIP5, BIRC5, AR. GO and KEGG analysis revealed that the network is involved in various biological functions. Immune infiltration analysis showed that IDD was closely related to immune cell infiltration, and hub genes could further affect the development of IDD by affecting immune cell infiltration. Conclusion: This study identified the hsa_circ_0074817-hsa-miR-508-5p-IGF1/CHEK1/CCNB1, the hsa_circ_0003600-hsa-miR-4741-BIRC5/OIP5/AR and the hsa_circ_0002702-hsa-miR-3158-5p-IGF1/AR as important regulatory axis of IDD, which will help us gain further insight into the pathogenesis of IDD and determine potential therapeutic targets. [ABSTRACT FROM AUTHOR]

Ferroptosis is a type of cell regulated necrosis triggered by intracellular phospholipid peroxidation, which is more immunogenic than apoptosis. Therefore, genes controlling ferroptosis may be promising candidate biomarkers for tumor therapy. In this study, we investigate the function of genes associated with ferroptosis in breast cancer (BC) and systematically evaluate the relationship between ferroptosis-related gene expression and prognosis of BC patients from the Cancer Genome Atlas database. By using the consensus clustering method, 1203 breast cancer samples were clustered into two clearly divided subgroups based on the expression of 237 ferroptosis-related genes. Then differentially expressed analysis and least absolute shrinkage and selection operator were used to identify the prognosis-related genes. Furthermore, the genetic risk signature was constructed using the expression of prognosis-related genes. Our results showed that the genetic risk signature can identify patient subgroups with distinct prognosis in either training cohort or validation, and the genetic risk signature was associated with the tumor immune microenvironment. Finally, the Cox regression analysis indicated that our risk signature was an independent prognostic factor for BC patients and this signature was verified by the polymerase chain reaction and western blot. Within this study, we identified a novel prognostic classifier based on five ferroptosis-related genes which may provide a new reference for the treatment of BRCA patients. [ABSTRACT FROM AUTHOR]

Background. Inflammatory reactions and pyroptosis play an important role in the pathology of intervertebral disc degeneration (IDD). The aim of the present study was to investigate pyroptosis in the nucleus pulposus cells (NPCs) of inflammatory induced IDD by bioinformatic methods and to search for possible diagnostic biomarkers. Methods. Gene expression profiles related to IDD were downloaded from the GEO database to identify differentially expressed genes (DEGs) between inflammation-induced IDD and non-inflammatory intervention samples. Pyroptosis genes were then searched for, and their expression in IDD was analyzed. Weighted gene co-expression network analysis (WGCNA) was then used to search for modules of IDD genes associated with pyroptosis and intersected with DEGs to discover candidate genes that would be diagnostically valuable. A LASSO model was developed to screen for genes that met the requirements, and ROC curves were created to clarify the diagnostic value of the genetic markers. Ultimately, the screened genes were further validated, and their diagnostic value assessed by selecting gene sets from the GEO database. RT-PCR was used to assess the mRNA expression of diagnostic markers in the nucleus pulposus (NP). Pan-cancer analysis was applied to demonstrate the expression and prognostic value of the screened genes in various tumors. Results. A total of 733 DEGs were identified in GSE41883 and GSE27494, which were mainly enriched in transmembrane receptor protein serine/threonine, kinase signaling pathway, response to lipopolysaccharide, and other biological processes, and they were mainly related to TGF beta signaling pathway, toll-like receptor signaling pathway, and TNF signaling pathway. A total of 81 genes related to pyroptosis were identified in the literature, and eight genes related to IDD were identified in the Veen diagram, namely, IL1A, IL1B, NOD2, GBP1, IL6, AK1, EEF2K, and PYCARD. Eleven candidate genes were obtained after locating the intersection of pyroptosis-related module genes and DEGs according to WGCNA analysis. A total of six valid genes were obtained after constructing a machine learning model, and five key genes were finally identified after correlation analysis. GSE23132 and GSE56081 validated the candidate genes, and the final IDD-related diagnostic markers were obtained as SMIM1 and SEZ6L2. RT-PCR results indicated that the mRNA expression of both was significantly elevated in IDD. The pan-cancer analysis demonstrated that SMIM1 and SEZ6L2 have important roles in the expression and prognosis of various tumors. Conclusion. In conclusion, this research identifies SMIM1 and SEZ6L2 as important biomarkers of IDD associated with pyroptosis, which will help to unravel the development and pathogenesis of IDD and determine potential therapeutic targets. [ABSTRACT FROM AUTHOR]

Polycystic ovary syndrome (PCOS) is a prevalent endocrine and metabolic disorder in premenopausal women. This investigation was to elucidate the underlying mechanism of endoplasmic reticulum stress (ERS) activation in granulosa cells, which has been implicated in the etiology of PCOS. Differentially expressed genes (DEGs) between PCOS and control groups were integrated with ERS gene lists from databases to identify DE-ERS genes, and functional analyses were performed. Univariate regression analysis and the LASSO method were used to select diagnostic factors, followed by establishing a DE-ERS gene-based diagnostic model. A nomogram model was further generated to predict the risk of PCOS. The correlation between ERS gene expression and immune cell proportion was assessed. A total of 14 DE-ERS genes associated with “protein processing in endoplasmic reticulum”, “ferroptosis”, and “glycerophospholipid metabolism” were selected as PCOS-related factors. An eight-DE-ERS genes-based diagnostic model was developed and displayed satisfactory performance in the training (Area under curve (AUC) = 0.983) and validation datasets (AUC = 0.802). High risk of PCOS can be accurately predicted, which might contribute to clinical decision-making. Moreover, EDEM1 expression was significantly positively correlated with naive B cell infiltration, while PDIA6 was negatively correlated with neutrophil proportion (P < 0.001). We identified eight novel molecules and developed an ERS gene-based diagnostic model in PCOS, which might provide novel insight for finding biomarkers and treatment methods. [ABSTRACT FROM AUTHOR]

The onset and progression of ovarian cancer (OC) are closely related to dysregulated gene expression. Current treatments for OC are mainly limited to surgery and chemotherapy. However, due to low drug sensitivity, the prognosis OC is exceptionally poor and the recurrence rate remains high. Hence, it is vital to develop new treatment strategies. Gene editing for site-specific genomic modification is a powerful novel tool for the treatment of OC. In this article, current gene editing research for the treatment of OC is reviewed to provide a reference for the clinical application of new approaches to improve treatment outcomes and prognosis. [ABSTRACT FROM AUTHOR]

This study aimed to analyze the role of the FcepsilonRI (FcεRI) signaling pathway in the pathogenesis of benign lymphoepithelial lesion of lacrimal gland (LGBLEL). Transcriptomic analysis was performed on LGBLEL and orbital cavernous hemangioma (CH) patients diagnosed via histopathology in Beijing Tongren Hospital, Capital Medical University, between July 2010 and October 2013. Four LGBLEL and three orbital CH patients, diagnosed between October 2018 and August 2019, were randomly selected as experimental and control groups, respectively. RT-PCR, immunohistochemical staining, and western blotting were used to verify genes and proteins related to the FcεRI signaling pathway. Transcriptomic analysis showed that the FcεRI signaling pathway was upregulated in the LGBLEL compared with the CH group. The mRNA expression levels of important genes including SYK, p38, JNK, PI3K, and ERK were significantly increased in the LGBLEL group (P = 0.0066, P = 0.0002, P = 0.0003, P < 0.0001, P < 0.0001, respectively). Immunohistochemical staining results showed that SYK, p38, and ERK were positively expressed in LGBLEL, while JNK and PI3K were not. The protein contents of P-SYK, P-p38, P-JNK, P-PI3K, and P-ERK were significantly higher in the LGBLEL than in the CH group (P = 0.0169, P = 0.0074, P = 0.0046, P = 0.0157, P = 0.0156, respectively). The FcεRI signaling pathway participates in the pathogenesis of LGBLEL. [ABSTRACT FROM AUTHOR]

The aim of this study was to explore the effects of the potential of vitamin C in relieving lead‐induced antioxidant and immune responses in Grass Carp Ctenopharyngodon idella kidney (CIK) cells. In this study, CIK cells were treated with vitamin C (0, 60, 80, 100, 120, and 140 µM) and lead (0 and 25 µM). Cell viability, oxidative response, and immune‐related gene expression were assessed. These results showed that cell viability was significantly decreased by exposure to lead and that vitamin C supplementation alone had no significant effect on cell viability, but vitamin C supplementation can alleviate the effect of lead on cell viability. Furthermore, vitamin C supplementation can increase the antioxidant capacity and the expression of immune‐related genes. Ultimately, these results suggest that vitamin C has a protective effect on CIK cells when they are exposed to lead. [ABSTRACT FROM AUTHOR]

Background: Natural variations derived from both evolutionary selection and genetic recombination, presume to have important functions to respond to various abiotic stresses, which could be used to improve drought tolerance via genomic selection. Results: In the present study, the NAC-encoding gene of ZmNAC080308 was cloned and sequenced in 199 inbred lines in maize. Phylogenetic analysis showed that ZmNAC080308 is closely clusteredinto the same group with other well-known NAC genes responding to improve drought tolerance. In total, 86 SNPs and 47 InDels were identified in the generic region of ZmNAC080308, 19 of these variations were associated with GY (grain yield) in different environments. Nine variations in the 5'-UTR region of ZmNAC080308 are closely linked, they might regulate the gene expression and respond to improve GY under drought condition via Sp1-mediated transactivation. Two haplotypes (Hap1 and Hap2) identified in the, 5'-UTR region using the nine variations, and Hap2 containing insertion variants, exhibited 15.47 % higher GY under drought stress condition. Further, a functional marker was developed to predict the drought stress tolerance in a US maize inbred line panel. Lines carrying Hap2 exhibited > 10 % higher GY than those carrying Hap1 under drought stress condition. In Arabidopsis, overexpression ZmNAC080308 enhanced drought tolerance. Conclusions: ZmNAC080308 is an important gene responding to drought tolerance, a functional marker is developed for improving maize drought tolerance by selecting this gene. [ABSTRACT FROM AUTHOR]

SUMMARY: Cold stress has always been a major abiotic factor affecting the yield and quality of temperate fruit crops. Ethylene plays a critical regulatory role in the cold stress response, but the underlying molecular mechanisms remain elusive. Here, we revealed that ethylene positively modulates apple responses to cold stress. Treatment with 1‐aminocyclopropane‐1‐carboxylate (an ethylene precursor) and aminoethoxyvinylglycine (an ethylene biosynthesis inhibitor) respectively increased and decreased the cold tolerance of apple seedlings. Consistent with the positive effects of ethylene on cold stress responses, a low‐temperature treatment rapidly induced ethylene release and the expression of MdERF1B, which encodes an ethylene signaling activator, in apple seedlings. Overexpression of MdERF1B significantly increased the cold tolerance of apple plant materials (seedlings and calli) and Arabidopsis thaliana seedlings. A quantitative real‐time PCR analysis indicated that MdERF1B upregulates the expression of the cold‐responsive gene MdCBF1 in apple seedlings. Moreover, MdCIbHLH1, which functions upstream of CBF‐dependent pathways, enhanced the binding of MdERF1B to target gene promoters as well as the consequent transcriptional activation. The stability of MdERF1B–MdCIbHLH1 was affected by cold stress and ethylene. Furthermore, MdERF1B interacted with the promoters of two genes critical for ethylene biosynthesis, MdACO1 and MdERF3. The resulting upregulated expression of these genes promoted ethylene production. However, the downregulated MdCIbHLH1 expression in MdERF1B‐overexpressing apple calli significantly inhibited ethylene production. These findings imply that MdERF1B–MdCIbHLH1 is a potential regulatory module that integrates the cold and ethylene signaling pathways in apple. Significance Statement: Here, we show that the ethylene response factor MdERF1B acting in the CBF/DREB1 signaling pathway was upregulated by cold treatment. Further investigation revealed that MdCIbHLH1, which functions upstream of CBF‐regulated pathways, is essential for MdERF1B‐mediated cold tolerance and ethylene biosynthesis. [ABSTRACT FROM AUTHOR]

Interleukin-1 receptor antagonist (IL-1Ra) is an antagonist of IL-1β binding IL-1β receptors but does not induce intracellular responses or signal transduction. In this study, the full-length complementary DNA (cDNA) of the IL-1Ra gene (OaIL-1Ra) was identified from sheep (Ovis aries) using rapid amplification of cDNA ends PCR and submitted to GenBank with the accession number KC425613. The OaIL-1Ra cDNA comprised an open reading frame of 525 bp encoding a protein of 19765.8 Da, a 5′-untranslated region (UTR) of 27 bp, and a 3′-UTR of 676 bp with a poly(A) tail. Recombinant OaIL-1Ra with bioactivity was expressed in a prokaryotic expression system, and a monoclonal antibody against native OaIL-1Ra was prepared. Through Western blot analyses, the OaIL-1Ra protein was widely expressed in lung, heart, spleen, liver, kidney, muscle, intestine, lymphonodi, rumen, and white blood cells, with the highest levels in liver and spleen. The expression of OaIL-1Ra in primary cultured white blood cells of sheep were highly induced in a time-dependent manner when challenged with different bacteria. These results implied that OaIL-1Ra is associated with immune responses during bacterial infections. [ABSTRACT FROM AUTHOR]

Background: Turner syndrome (TS) is a sex chromosome aneuploidy with a variable spectrum of symptoms including short stature, ovarian failure and skeletal abnormalities. The etiology of TS is complex, and the mechanisms driving its pathogenesis remain unclear. Methods: In our study, we used the online Gene Expression Omnibus (GEO) microarray expression profiling dataset GSE46687 to identify differentially expressed genes (DEGs) between monosomy X TS patients and normal female individuals. The relevant data on 26 subjects with TS (45,XO) and 10 subjects with the normal karyotype (46,XX) was investigated. Then, tissue-specific gene expression, functional enrichment, and protein-protein interaction (PPI) network analyses were performed, and the key modules were identified. Results: In total, 25 upregulated and 60 downregulated genes were identified in the differential expression analysis. The tissue-specific gene expression analysis of the DEGs revealed that the system with the most highly enriched tissue-specific gene expression was the hematologic/immune system, followed by the skin/skeletal muscle and neurologic systems. The PPI network analysis, construction of key modules and manual screening of tissue-specific gene expression resulted in the identification of the following five genes of interest: CD99, CSF2RA, MYL9, MYLPF , and IGFBP2. CD99 and CSF2RA are involved in the hematologic/immune system, MYL9 and MYLPF are related to the circulatory system, and IGFBP2 is related to skeletal abnormalities. In addition, several genes of interest with possible roles in the pathogenesis of TS were identified as being associated with the hematologic/immune system or metabolism. Conclusion: This discovery-driven analysis may be a useful method for elucidating novel mechanisms underlying TS. However, more experiments are needed to further explore the relationships between these genes and TS in the future. [ABSTRACT FROM AUTHOR]

Anthocyanin biosynthesis is induced by cytokinins, and is regulated by MYB transcription factors. However, the underlying molecular mechanisms have not been fully characterised. In the present study, red-fleshed apple callus were induced from the leaves of an R6/R6 homozygous line, which was the hybrid offspring of Malus sieversii f. niedzwetzkyana and 'Fuji'. We analysed the callus anthocyanin contents in response to different cytokinin concentrations. We observed that cytokinin treatments upregulated the expression of anthocyanin structural genes MdDFR and MdUFGT and transcription factor genes MdMYB10 and MdbHLH3. Additionally, the expression of MdMYBL2 , which encodes the bHLH and EAR motifs, was inhibited by cytokinin treatments. The MdMYBL2 -overexpressing callus had lower anthocyanin contents than the wild-type controls. We noted that the expression levels of anthocyanin biosynthesis structural genes MdDFR and MdUFGT and transcription factor genes MdMYB10 and MdbHLH3 were strongly suppressed in the transgenic callus. Subsequent yeast two-hybrid, bimolecular fluorescence complementation, and pull-down assays indicated that MdMYBL2 interacts with MdbHLH3, which may influence the expression of anthocyanin biosynthesis-related genes. Our findings may provide new insights into how MYB transcription factors influence the cytokinin-regulated anthocyanin biosynthesis in red-fleshed apples. During the cultivation of fruit trees, the rational application of hormones based on different environmental conditions can improve the appearance and nutritional value of fruit crops. Herein, we discussed the involvement of cytokinin in the anthocyanin biosynthesis pathway, which may have implications for the development of new cultivation techniques aimed at improving fruit coloration under diverse environmental conditions. [ABSTRACT FROM AUTHOR]

T‐2 toxin is one of the trichothecene mycotoxins commonly found in animal feed and agricultural products. Ingestion of T‐2 toxin by animals results in acute and chronic diseases, as well as reproductive failure. This study aimed at investigating the role of endoplasmic reticulum (ER) stress in T‐2 toxin‐induced apoptosis in goat endometrium epithelial cells. Flow cytometry and cell viability assay showed that T‐2 toxin significantly induced cell apoptosis, which was accompanied with increased expression of cleaved‐caspase‐3. The altered expression of two ER stress markers CHOP and GRP78 proved that ER stress is involved in the T‐2 toxin‐induced apoptosis. 4‐phenylbutyrate pretreatment was applied to relieve the ER stress, pretreated endometrium epithelial cells showed a decreased apoptosis level and the expression pattern of CHOP and GRP78 was reversed. The key genes involved in signaling pathways of ER stress‐associated apoptosis were examined, which showed that the IRE1‐JNK and PERK‐ATF4‐CHOP signal transduction pathways are both activated. Moreover, the level of cytokines including interleukin (IL)‐1β, IL‐6, IL‐10 and tumor necrosis factor‐α decreased in the T‐2 toxin‐treated cells. While the 4‐phenylbutyrate pretreatment elevates the cytokine levels after T‐2 treatment. Collectively, these results suggest that ER stress contributes to the T‐2 toxin‐induced apoptosis and decreased cytokine levels in goat endometrium epithelial cells. This study offers new insight into the molecular mechanisms of T‐2 toxicity on reproductive cells. T‐2 toxin‐induced apoptosis in goat endometrium epithelial cells is accompanied with increased expression of cleaved‐caspase‐3, and altered expression of CHOP and Grp78. The ER stress inhibitor, 4‐phenylbutyrate, decreased the apoptosis level and reversed the expression pattern of CHOP and GRP78 caused by T‐2 toxins. The IRE1‐JNK and PERK‐ATF4‐CHOP signal transduction pathways are both activated during T‐2 toxin exposure. These results indicated that ER stress is involved in T‐2 toxin‐induced apoptosis, which offers new insight into the molecular mechanisms of T‐2 toxicity on endometrium epithelial cells. [ABSTRACT FROM AUTHOR]

Introduction: Specific immunotherapy is critical for alleviating symptoms associated with house dust mite allergy, such as asthma and rhinitis. However, this approach relies on crude extracts, which are often not of sufficient quality or purity and are not standardized. The use of recombinant allergens may enable safer, more effective treatment.Material and Methods: Using our previously constructed plasmids pET28a(+)-Der f 1, pET28a(+)-Der f 2 and pET28b(+)-Der f 4 as templates, the gene fragments coding for the allergens Der f 1, Der f 2 and Der f 4, respectively, of the dust mite Dermatophagoides farinae were amplified by PCR. Next the PCR-amplified DNAs were recovered, cloned into pFastBacHT A, and transformed into Escherichia coli DH10Bac. The resulting vectors were co-transfected into Spodoptera frugiperda Sf9 cells for expression. The recombinant allergens were purified by Ni2+ affinity chromatography, and identified by SDS-PAGE and ELISA.Results: The recombinant allergens were successfully expressed and purified from a baculovirus expression system introduced into Sf9 cells, which were verified as being of the correct predicted molecular weights by SDS-PAGE. Furthermore, the reactivity to recombinant allergens rDer f 1, rDer f 2, and rDer f 4 was 85.2%, 88.9%, and 44.4%, respectively, in 27 children with asthma and D. farinae allergy.Conclusions: Recombinant allergens from dust mites can be successfully generated using a baculovirus-insect expression system. Furthermore, these recombinant allergens can be used to detect mite sensitivity in sera, highlighting their utility in future work to understand and develop treatment for mite allergy. [ABSTRACT FROM AUTHOR]

Purpose: A series of microRNAs (miRNAs) have been identified as associated with the survival of patients with non-small-cell lung cancer (NSCLC). The aim of the present study was to explore whether combination of these experimentally validated individual miRNA biomarkers could be used to further increase their prognostic power in NSCLC.Patients and methods: Based on previously validated NSCLC prognostic miRNAs, gene signatures that could discriminate high-risk subgroups with poor clinical outcome in four NSCLC miRNA expression datasets (GSE13937, GSE16025, The Cancer Genome Atlas Lung Adenocarcinoma, and TCGA lung squamous cell carcinoma) were developed using the SurvMicro tool. The potential of the miRNA signature established was validated by quantitative real-time PCR analysis of clinical NSCLC samples, and its prognostic power evaluated using the survivalROC method.Results: We developed two miRNA signatures with prognostic significance for NSCLC, comprising 12- and 7-miRNAs. The 7-miRNA signature (miR-148b, miR-365, miR-32, miR-375, miR-21, miR-125b, and miR-155) was a subset of a 12-miRNA set that retained prognostic power across NSCLC cohorts. Compared with previously established miRNA signatures, our 7-miRNA signature has similar potential, while comprising fewer miRNA components. The prognostic ability of the 7-miRNA signature was validated experimentally in an independent NSCLC cohort using real-time PCR (HR=3.4847, 95% CI=1.3693–8.8680, P=0.0092), and this signature, combined with tumor pathological stage, had superior prognostic ability compared with tumor stage alone.Conclusion: Our data indicate that the established 7-miRNA signature is simple, robust, and may have greater clinical prognostic utility for patients with NSCLC. [ABSTRACT FROM AUTHOR]