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Striatin-interacting protein 1 (Strip1) is a core component of the striatin interacting phosphatase and kinase (STRIPAK) complex, which is involved in embryogenesis and development, circadian rhythms, type 2 diabetes, and cancer progression. However, the expression and role of Strip1 in the mammalian cochlea remains unclear. Here we studied the expression and function of Strip1 in the mouse cochlea by using Strip1 knockout mice. We first found that the mRNA and protein expression of Strip1 increases as mice age starting from postnatal day (P) 3 and reaches its highest expression level at P30 and that the expression of Strip1 can be detected by immunofluorescent staining starting from P14 only in cochlear HCs, and not in supporting cells (SCs). Next, we crossed Strip1 heterozygous knockout (Strip +/−) mice to obtain Strip1 homozygous knockout (Strip1 −/−) mice for studying the role of Strip1 in cochlear HCs. However, no Strip1 −/− mice were obtained and the ratio of Strip +/− to Strip1 +/+ mice per litter was about 2:1, which suggested that homozygous Strip1 knockout is embryonic lethal. We measured hearing function and counted the HC number in P30 and P60 Strip +/− mice and found that they had normal hearing ability and HC numbers compared to Strip1 +/+ mice. Our study suggested that Strip1 probably play important roles in HC development and maturation, which needs further study in the future. [ABSTRACT FROM AUTHOR]

Introduction/Background: Global warming greatly limits the productivity of rice. Rice plants are highly sensitive to heat stress at the flowering stage. The selection of heat-tolerant varieties is considered the most effective approach for ensuring global food security in the coming decades. Methods: Based on previous screening and QTL localization results, we selected tolerant varieties (Huang Huazhan, HZ) and susceptible varieties (Yang Dao6, YD) of rice and studied their physiological characteristics, gene expression responses, and proteomic differences of their anthers under heat stress. The differentially expressed proteins (DEPs) were validated by real-time PCR. Results: The activities of the antioxidant enzymes CAT, SOD, POD, and APX were 8.36%, 9.56%, 20.61%, and 25.34% higher in HZ than in YD under heat stress, respectively. Similarly, the content of proline and soluble sugar was 8.32% and 14.47% higher in HZ than in YD, respectively. The content of MDA and H2O2 was 8.11% and 39.5% lower in HZ than in YD, respectively. The ratio of endogenous GA3/ABA in HZ was 10.65, which was significantly higher than that of YD (3.84). In addition, we validated the candidate genes LOC_Os08g07010 and LOC_Os08g07440 that our team located in 2021, and the result showed that the expression of these two heat-tolerant genes in the anthers was significantly higher in HH than in YH. DEPs involved in the response to heat stress were identified by TMT proteomics, five upregulated and three downregulated differential expression proteins in HH. DEPs were verified by RT-qPCR. Discussion: These results provide new insights into the physiological characteristics, dominant DEPs, and gene expression responses in both rice varieties under heat stress. Our results indicate that the antioxidant and osmoregulatory capacities, the ratio of endogenous GA3 and ABA, these DEPs are mainly involved in the pathways of phenylpropanoid biosynthesis, ubiquitin-mediated proteolysis, carbohydrate metabolism, thiamine metabolism, protein processing in the endoplasmic reticulum, and folding, sorting, and degradation were upregulated to a greater degree in HZ than in YD. Additional studies were performed to clarify the roles of these proteins in response to heat stress. [ABSTRACT FROM AUTHOR]

The study of long noncoding RNAs (lncRNAs) is an emerging area of cancer research, in part due to their ability to serve as disease biomarkers. However, few studies have investigated lncRNAs in chronic lymphocytic leukemia (CLL). We have identified one particular lncRNA, treRNA, which is overexpressed in CLL B-cells. We measured transcript expression in 144 CLL patient samples and separated samples into high or low expression of treRNA relative to the overall median. We found that high expression of treRNA is significantly associated with shorter time to treatment. High treRNA also correlates with poor prognostic indicators such as unmutated IGHV and high ZAP70 protein expression. We validated these initial findings in samples collected in a clinical trial comparing the nucleoside analog fludarabine alone or in combination with the alkylating agent cyclophosphamide in untreated CLL samples collected prior to starting therapy (E2997). High expression of treRNA was independently prognostic for shorter progression free survival in patients receiving fludarabine plus cyclophosphamide. Given these results, in order to study the role of treRNA in DNA damage response we generated a model cell line system where treRNA was over-expressed in the human B-CLL cell line OSU-CLL. Relative to the vector control line, there was less cell death in OSU-CLL over-expressing treRNA after exposure to fludarabine and mafosfamide, due in part to a reduction in DNA damage. Therefore, we suggest that treRNA is a novel biomarker in CLL associated with aggressive disease and poor response to chemotherapy through enhanced protection against cytotoxic mediated DNA damage.

Parental genes can influence the phenotype of their offspring through genomic-epigenomic interactions even without the direct inheritance of specific parental genotypes. Maternal genetic variations can affect the ovarian and intrauterine environments and potentially alter lactation behaviors, impacting offspring nutrition and health outcomes independently of the fetal genome. Similarly, paternal genetic changes can affect the endocrine system and vascular functions in the testes, influencing sperm quality and seminal fluid composition. These changes can initiate early epigenetic modifications in sperm, including alterations in microRNAs, tRNA-derived small RNAs (tsRNAs), and DNA methylation patterns. These epigenetic modifications might induce further changes in target organs of the offspring, leading to modified gene expression and phenotypic outcomes without transmitting the original parental genetic alterations. This review presents clinical evidence supporting this hypothesis and discusses the potential underlying molecular mechanisms. Parental gene-offspring epigenome-offspring phenotype interactions have been observed in neurocognitive disorders and cardio-renal diseases. [ABSTRACT FROM AUTHOR]

Few reports have confirmed whether exosomes derived from fibroblasts can regulate the process of melanogenesis. We wondered whether exosomes derived from fibroblasts could have a potent regulatory effect on melanogenesis and explored the underlying mechanisms. This study aimed to find the role of fibroblasts in melanocytes and revealed the related mechanisms. RT-qPCR, Western blot analysis were conducted to measure the RNA and protein expression level of various related genes. miRNA sequencing, mass spectrum analysis and subsequent bioinformatics analysis were employed to find the underlying targets. Zebrafish were employed to measure the melanin synthesis related process in vivo. Furthermore, electron microscopy, ROS measurement and dual-luciferase reporter assay were adopted to investigate the relationship between these processes. We found that exosomes derived from human primary dermal fibroblasts were internalized by human primary melanocytes and MNT1 cells and that the melanin content and the expression of melanin synthesis-related proteins TYR and MITF was inhibited by exosomes derived from UVB-induced human primary dermal fibroblasts. The miRNA expression profile in secreted exosomes changed significantly, with miR-25–5p identified as capable of regulating TSC2 expression via the CDS region. The miR-25–5p-TSC2 axis could affect the melanin content through subsequent cellular organelle dysfunction, such as mitochondrial dysfunction, endoplasmic reticulum stress and dysregulation of lysosomal cysteine proteases. We unveiled a novel regulatory role of fibroblasts in melanocytes, facilitated by the secretion of exosomes. miR-25–5p within exosomes plays a pivotal role in regulating melanogenesis via TSC2-induced cellular organelle dysfunction. • A new regulatory role of fibroblasts in melanocytes was revealed and this process was achieved by the secretion of exosomes. • miR-25–5p in exosomes plays a role in regulating melanogenesis. • TSC2-induced cellular organelle dysfunction could affect the process of melanogenesis. [ABSTRACT FROM AUTHOR]

Sclerostin (SOST) is produced by osteocytes and is known as a negative regulator of bone homeostasis. Parathyroid hormone (PTH) regulates calcium, phosphate as well as vitamin D metabolism, and is a strong inhibitor of SOST synthesis in vitro and in vivo. PTH has two methionine amino acids (positions 8 and 18) which can be oxidized. PTH oxidized at Met18 (Met18(ox)-PTH) continues to be bioactive, whereas PTH oxidized at Met8 (Met8(ox)-PTH) or PTH oxidized at Met8 and Met18 (Met8, Met18(di-ox)-PTH) has minor bioactivity. How non-oxidized PTH (n-oxPTH) and oxidized forms of PTH act on sclerostin synthesis is unknown. The effects of n-oxPTH and oxidized forms of PTH on SOST gene expression were evaluated in UMR106 osteoblast-like cells. Moreover, we analyzed the relationship of SOST with n-oxPTH and all forms of oxPTH in 516 stable kidney transplant recipients using an assay system that can distinguish in clinical samples between n-oxPTH and the sum of all oxidized PTH forms (Met8(ox)-PTH, Met18(ox)-PTH, and Met8, Met18(di-ox)-PTH). We found that both n-oxPTH and Met18(ox)-PTH at doses of 1, 3, 20, and 30 nmol/L significantly inhibit SOST gene expression in vitro, whereas Met8(ox)-PTH and Met8, Met18(di-ox)-PTH only have a weak inhibitory effect on SOST gene expression. In the clinical cohort, multivariate linear regression showed that only n-oxPTH, but not intact PTH (iPTH) nor oxPTH, is independently associated with circulating SOST after adjusting for known confounding factors. In conclusion, only bioactive PTH forms such as n-oxPTH and Met18(ox)-PTH, inhibit SOST synthesis. [ABSTRACT FROM AUTHOR]

Circulating cfRNA in plasma has emerged as a fascinating area of research with potential applications in disease diagnosis, monitoring, and personalized medicine. Circulating RNA sequencing technology allows for the non-invasive collection of important information about the expression of target genes, eliminating the need for biopsies. This comprehensive review aims to provide a detailed overview of the current knowledge and advancements in the study of plasma cfRNA, focusing on its diverse landscape and biological functions, detection methods, its diagnostic and prognostic potential in various diseases, challenges, and future perspectives. [ABSTRACT FROM AUTHOR]

Numerous studies have been conducted to investigate the genomic characterization of bZIP genes and their involvement in the cellular response to endoplasmic reticulum (ER) stress. These studies have provided valuable insights into the coordinated cellular response to ER stress, which is mediated by bZIP transcription factors (TFs). However, a comprehensive and systematic investigations regarding the role of bZIP genes and their involvement in ER stress response in pak choi is currently lacking in the existing literature. To address this knowledge gap, the current study was initiated to elucidate the genomic characteristics of bZIP genes, gain insight into their expression patterns during ER stress in pak choi, and investigate the protein-to-protein interaction of bZIP genes with the ER chaperone BiP. In total, 112 members of the BcbZIP genes were identified through a comprehensive genome-wide analysis. Based on an analysis of sequence similarity, gene structure, conserved domains, and responsive motifs, the identified BcbZIP genes were categorized into 10 distinct subfamilies through phylogenetic analysis. Chromosomal location and duplication events provided insight into their genomic context and evolutionary history. Divergence analysis estimated their evolutionary history with a predicted divergence time ranging from 0.73 to 80.71 million years ago (MYA). Promoter regions of the BcbZIP genes were discovered to exhibit a wide variety of cis-elements, including light, hormone, and stress-responsive elements. GO enrichment analysis further confirmed their roles in the ER unfolded protein response (UPR), while co-expression network analysis showed a strong relationship of BcbZIP genes with ER-stress-responsive genes. Moreover, gene expression profiles and protein–protein interaction with ER chaperone BiP further confirmed their roles and capacity to respond to ER stress in pak choi. [ABSTRACT FROM AUTHOR]

Flower color is a major feature of ornamental plants, and the rich flower color of plants is an important factor in determining their ornamental and economic values, so flower color is an important research target for gardening and horticulture breeders at home and abroad. Our research group collected four colors of Impatiens uliginosa (white, pink, red, and deep red) during the collection of germplasm resources in the field. In this study, we analyzed the transcriptomes of the four flower colors of I. uliginosa by using RNA-Seq technology. The transcriptomes were screened to identify candidate genes related to flower color, and the coloring mechanisms of four flower colors were revealed at the molecular level. The main findings were as follows: (1) The number of the four different transcripts ranged from 64,723 to 93,522 and contained a total of 100,705 unigenes. (2) The analysis of differentially expressed genes revealed structural genes including C4H, FLS, PAL, and ANS and transcription factors including MYB, MYB-related, AP2-EREBP, and bHLH. (3) Among the four flower colors of I. uliginosa, the C4H1 gene had the highest expression in pink flowers, and the C4H2 gene had the highest expression in red flowers. This indicated that C4H genes positively regulated the red flower color of I. uliginosa. However, FLS expression was the highest in white flowers, and with deepening flower color, FLS gene expression gradually weakened, acting as a negative regulator. The results of this study could lay the theoretical foundation for investigating the mechanism of coloration and flower color variation in I. uliginosa. [ABSTRACT FROM AUTHOR]

Impatiens uliginosa is a native and potential water body-restoring ornamental plant. In this study, RNA-Seq technology was used to analyze the transcriptome of its floral organs. Candidate genes related to flower color changes under copper stress were investigated through transcriptome screening, and the intrinsic mechanism of the effects of different concentrations of copper on I. uliginosa was revealed at the molecular level. The main findings were as follows: (1) Transcriptome sequencing analysis was performed on the flower organs of I. uliginosa treated with different concentrations of copper (0 mg·L−1, 10 mg·L−1, and 20 mg·L−1). A total of 70,319 transcripts and 39,949 unigenes were obtained. An analysis of differentially expressed genes revealed structural genes including GT, ANS, CHI, and PAL; transcription factors including MYB and WD40; and transport factors including GST and ABC. (2) The gene expression levels of flower color changed in the flowering period of I. uliginosa at different copper concentrations. The expression levels of IuGT and IuGST genes in I. uliginosa were significantly different under different concentrations of copper treatments. Their expression levels were the highest at a copper concentration of 0 mg·L−1 and the lowest at 20 mg·L−1. In summary, the low expression of IuGT and IuGST genes was more conducive to the formation of white flowers of I. uliginosa. [ABSTRACT FROM AUTHOR]

Background: Hepatocellular carcinoma (HCC) is characterized by aggressive progression and elevated mortality rates. This study aimed to investigate the regulatory effects of RBBP7 on HCC pathogenesis and the underlying mechanisms. Methods: The expression and clinical feature of RBBP7 were evaluated using bioinformatics analysis and the assessment of clinical HCC samples. CCK8 and colony formation were employed to estimate cell proliferation function of RBBP7. Aerobic glycolysis levels of RBBP7 were evaluated by measuring ATP levels, lactic acid production, glucose uptake capacity, and the expression of relevant enzymes (PFKM, PKM2, and LDHA). The phosphorylation levels in PI3K/AKT signaling were measured by western blotting. The regulatory effect of transcription factors of specificity protein 1 (SP1) on RBBP7 mRNA expression was confirmed in dual-luciferase reporter assays and chromatin immunoprecipitation experiments. The proliferation- and glycolysis-associated proteins were assessed using immunofluorescence staining in vivo. Results: We found that RBBP7 is expressed at high levels in HCC and predicts poor survival. Functional assays showed that RBBP7 promoted HCC proliferation and glycolysis. Mechanistically, it was demonstrated that RBBP7 activates the PI3K/AKT pathway, a crucial pathway in glycolysis, contributing to the progression of HCC. The outcomes of the dual-luciferase assay further confirmed that SP1 is capable of activating the promoter of RBBP7. Conclusions: RBBP7, which is up-regulated by SP1, promotes HCC cell proliferation and glycolysis through the PI3K/AKT pathway. The findings of this study suggest that RBBP7 is a potential biomarker for HCC. [ABSTRACT FROM AUTHOR]

Background and aims: Amino acids (AAs) not only constitute milk protein but also stimulate milk synthesis through the activation of mTORC1 signaling, but which amino acids that have the greatest impact on milk fat and protein synthesis is still very limited. In this study, we aimed to identify the most critical AAs involved in the regulation of milk synthesis and clarify how these AAs regulate milk synthesis through the G-protein-coupled receptors (GPCRs) signaling pathway. Methods: In this study, a mouse mammary epithelial cell line (HC11) and porcine mammary epithelial cells (PMECs) were selected as study subjects. After treatment with different AAs, the amount of milk protein and milk fat synthesis were detected. Activation of mTORC1 and GPCRs signaling induced by AAs was also investigated. Results: In this study, we demonstrate that essential amino acids (EAAs) are crucial to promote lactation by increasing the expression of genes and proteins related to milk synthesis, such as ACACA, FABP4, DGAT1, SREBP1, α-casein, β-casein, and WAP in HC11 cells and PMECs. In addition to activating mTORC1, EAAs uniquely regulate the expression of calcium-sensing receptor (CaSR) among all amino-acid-responsive GPCRs, which indicates a potential link between CaSR and the mTORC1 pathway in mammary gland epithelial cells. Compared with other EAAs, leucine and arginine had the greatest capacity to trigger GPCRs (p-ERK) and mTORC1 (p-S6K1) signaling in HC11 cells. In addition, CaSR and its downstream G proteins Gi, Gq, and Gβγ are involved in the regulation of leucine- and arginine-induced milk synthesis and mTORC1 activation. Taken together, our data suggest that leucine and arginine can efficiently trigger milk synthesis through the CaSR/Gi/mTORC1 and CaSR/Gq/mTORC1 pathways. Conclusion: We found that the G-protein-coupled receptor CaSR is an important amino acid sensor in mammary epithelial cells. Leucine and arginine promote milk synthesis partially through the CaSR/Gi/mTORC1 and CaSR/Gq/mTORC1 signaling systems in mammary gland epithelial cells. Although this mechanism needs further verification, it is foreseeable that this mechanism may provide new insights into the regulation of milk synthesis. [ABSTRACT FROM AUTHOR]

Cauliflower is a nutritious vegetable with inflorescences that are specialized to form the edible organs called curds. Uncovering key genes underlying important traits is crucial for the genetic improvement of this important crop. However, the genetic basis of many important agronomic traits, including curd performance and plant architecture in cauliflower, remains unclear. GWASs have proved to be powerful tools to study agronomic traits in many crops. To reveal the genetic basis of four important agronomic traits, namely, the main stem height (MSH), purplish curd (PC), external leaf wing (ELW) and weight of a single curd (WSC), we selected 220 core accessions of loose-curd cauliflower for resequencing, phenotypic investigation and GWAS. The approach revealed significant novel loci. We detected several significant associations: on C02 for MSH and PC, on C06 for ELW and on C01 for WSC. More interestingly, we identified a significant single-peak signal for the weight of a single curd (WSC), an important yield trait, and within this signal interval, we identified the BOB01G136670 gene with five SNPs encoding nonsynonymous mutations in the CDS region; these mutations resulted in two haplotypes with significant differences in curd weight. The weight of a single curd was significantly increased in the varieties with the BOB01G136670 Hap1 allele compared to those with BOB01G136670 Hap2. BOB01G136670 was highly conserved with the homologous genes that encode serine carboxypeptidase and belong to the S10 family in other species, including GS5, which functions as a positive regulator of grain size in rice, wheat and maize. Additionally, BOB01G136670 was highly expressed specifically at the curd enlargement stage, with low or even no expression at all in other tissues and stages, indicating that BOB01G136670 is a plausible candidate gene for WSC. Overall, this study identified genomic loci for four important agronomic traits that are relevant for accelerating biological breeding and the improvement of cauliflower varieties. [ABSTRACT FROM AUTHOR]

Poly (ADP-ribose) Polymerase (PARP) inhibitors (PARPi) have been approved for both frontline and recurrent setting in ovarian cancer with homologous recombination (HR) repair deficiency. However, more than 40% of BRCA1/2-mutated ovarian cancer lack the initial response to PARPi treatment, and the majority of those that initially respond eventually develop resistance. Our previous study has demonstrated that increased expression of aldehyde dehydrogenase 1A1 (ALDH1A1) contributes to PARPi resistance in BRCA2-mutated ovarian cancer cells by enhancing microhomology-mediated end joining (MMEJ) but the mechanism remains unknown. Here, we find that ALDH1A1 enhances the expression of DNA polymerase θ (Polθ, encoded by the POLQ gene) in ovarian cancer cells. Furthermore, we demonstrate that the retinoic acid (RA) pathway is involved in the transcription activation of the POLQ gene. The RA receptor (RAR) can bind to the retinoic acid response element (RARE) located in the promoter of the POLQ gene, promoting transcription activation-related histone modification in the presence of RA. Given that ALDH1A1 catalyzes the biosynthesis of RA, we conclude that ALDH1A1 promotes POLQ expression via the activation of the RA signaling pathway. Finally, using a clinically-relevant patient-derived organoid (PDO) model, we find that ALDH1A1 inhibition by the pharmacological inhibitor NCT-505 in combination with the PARP inhibitor olaparib synergistically reduce the cell viability of PDOs carrying BRCA1/2 mutation and positive ALDH1A1 expression. In summary, our study elucidates a new mechanism contributing to PARPi resistance in HR-deficient ovarian cancer and shows the therapeutic potential of combining PARPi and ALDH1A1 inhibition in treating these patients. [ABSTRACT FROM AUTHOR]

BACKGROUND: Lymphoid-specific helicase (HELLS), a SNF2-like chromatin-remodeling enzyme, plays a key role in tumor progression via its DNA methylation function. However, the effects of HELLS on immune infiltration and prognosis in liver hepatocellular carcinoma (LIHC) remain uncertain. METHODS: The Tumor Immune Estimation Resource (TIMER) database was employed to explore the pan-cancer mRNA expression of HELLS and its correlation with immunity. GEPIA2 was used to verify the correlation between HELLS expression and survival. The role of HELLS in cancer was explored via gene set enrichment analysis (Gene Ontology and Kyoto Encyclopedia of Genes and Genomes) and the construction of gene-gene and protein-protein interaction networks (PPI). Additionally, correlations between DNA methylation, HELLS expression, and immune-related genes were explored in LIHC. HELLS expression in LIHC clinical samples was determined using qRT-PCR and western blotting. The effects of downregulated HELLS expression in hepatocellular carcinoma cells was explored via transfection experiments in vitro. RESULTS: High HELLS mRNA expression was identified in several cancers and was significantly associated with poorer prognosis in LIHC. Furthermore, HELLS expression was positively correlated with tumor-infiltrating lymphocytes and immune checkpoint genes in LIHC. Bioinformatics analysis suggested that DNA methylation of HELLS may be associated with the immune response. Results from the TCGA-LIHC dataset, clinical samples, and functional analysis indicated that HELLS contributed to tumor progression in LIHC. CONCLUSION: The study findings demonstrate that HELLS is an important factor in promoting LIHC malignancy and might serve as a potential biomarker for LIHC. [ABSTRACT FROM AUTHOR]

Radish (Raphanus sativus L.) is an important root vegetable crop that is easily infected by various pathogens that result in decreased yield and quality. Nucleotide-binding site and leucine-rich repeat (NBS-LRR) genes play vital roles in resisting pathogen infection in plants. However, the genome-wide characterization and functional roles of NBS-LRR genes remain largely unexplored in radish. Here, a total of 187 RsNBS-LRR genes were identified at the whole-genome level in radish, among which 80 RsNBS-LRR genes were unevenly distributed on nine radish chromosomes. Interestingly, 15 clusters containing 36 RsNBS-LRR genes occurred in eight chromosomes. RNA-Seq data showed that several RsNBS-LRR genes exhibited significant differential expression profiles in different radish tissues. Moreover, a range of cis-acting regulatory elements associated with ABA, MeJA, or SA were identified in the promoter region of some RsNBS-LRR genes. RT-qPCR analysis showed that the expression of a few RsNBS-LRR genes (e.g., RsNBS021 and RsNBS163) was significantly induced under Peronospora parasitica infection and/or ABA treatment, indicating that they might play critical roles in ABA-dependent defense resistance processes. These results could enhance our understanding of the evolutionary relationship of RsNBS-LRR genes and facilitate the genetic manipulation of disease resistance in radish breeding programs. [ABSTRACT FROM AUTHOR]

In most higher plants, sucrose is the significant form of carbohydrate for long-distance transportation. Sucrose transporters/sucrose carriers (SUTs/SUCs) are involved in the loading and unloading of sucrose in phloem and play an important role in the growth and development of plants. In this study, 12 RsSUC genes were first identified from the radish genome, and their phylogenetic relationships, gene structure, and conserved motifs were further analyzed. RT-qPCR results indicated that RsSUC genes exhibited various expression patterns in different tissues and development stages of the radish. Overexpression of RsSUC1b in Arabidopsis significantly improved the uptake efficiency of exogenous sucrose, and promoted leaves and lateral root growth. In addition, the transgenic plants flowered significantly earlier than wild-type (WT) plants, and the soluble sugar contents (SSCs) including sucrose, glucose, and fructose in the mature leaves and pods were increased. It could be inferred that RsSUC1b is a plasma membrane sucrose transporter and plays a vital role in sucrose transportation and sugar accumulation during plant growth and development. These findings provided novel insights into the biological function of RsSUC genes and facilitate dissecting the molecular mechanism underlying sugar transport during radish development. [ABSTRACT FROM AUTHOR]

The late embryonic development abundant protein (LEA) is a family of proteins widely present in the body and related to osmoregulation. Saussurea involucrata is an extremely cold-tolerant plant. In our previous studies, we found that the LEAs gene in Saussurea involucrata has up-regulated expression under low temperature. To evaluate the biological function of SiLEA5 protein under low-temperature stress and its potential in agricultural breeding, we isolated the SiLEA5 gene from Saussurea involucrata, constructed a plant overexpression vector, and transformed tomato. We found that SiLEA5 protein significantly increased the yield of transgenic tomatoes by increasing their photosynthetic capacity, including net photosynthetic rate, stomatal conductance, and intercellular CO2 concentration. Under low-temperature stress, the SiLEA5 protein can regulate proline metabolism and oxidative stress, which confers transgenic tomatos with cold resistance. Thus, our work provided evidence for the role of SiLEA5 protein in low-temperature stress resistance in plants, as well as potential applications in crop breeding and cold stress resistance research. [ABSTRACT FROM AUTHOR]

Breast cancer is one of the most prevalent malignancies among women. Enhancing the prognosis is an effective approach to enhance the survival rate of breast cancer. Cuproptosis, a copper-dependent programmed cell death process, has been associated with patient prognosis. Inducing cuproptosis is a promising approach for therapy. However, there is currently no anti-breast cancer drug that induces cuproptosis. In this study, we repositioned the clinical drug fluphenazine as a potential agent for breast cancer treatment by inducing cuproptosis. Firstly, we utilized the Cancer Genome Atlas (TCGA) database and Connectivity Map (CMap) database to identify 22 potential compounds with anti-breast cancer activity through inducing cuproptosis. Subsequently, our findings demonstrated that fluphenazine effectively suppressed the viability of MCF-7 cells. Fluphenazine also significantly inhibited the viability of triple negative breast cancer cells MDA-MB-453 and MDA-MB-231. Furthermore, our study revealed that fluphenazine significantly down-regulated the expression of potential prognostic biomarkers associated with cuproptosis, increased copper ion levels, and reduced intracellular pyruvate accumulation. Additionally, it up-regulated the expression of FDX1 at both the mRNA and protein levels, which has been reported to play a crucial role in the induction of cuproptosis. These findings suggest that fluphenazine has the potential to be used as an anti-breast cancer drug by inducing cuproptosis. Therefore, this research provides an insight for the development of novel cuproptosis-dependent anti-cancer agents. [Display omitted] • The upregulation of SLC31A1 , ATP7A , and DLD is related to a poor prognosis in breast cancer. • Repositioning Flu as a potential drug candidate for the treatment of breast cancer. • The expression of SLC31A1 , ATP7A , and DLD was downregulated by Flu. • The anti-breast cancer effect of Flu is achieved by inducing cuproptosis. [ABSTRACT FROM AUTHOR]

Simple Summary: Estrogen receptors α (ERα) and β (ERβ) show distinct contributions to tumor initiation and progression. Given that ERβ mainly functions as a "tumor suppressor", activation of ERβ using its specific agonist should be able to inhibit tumor progression. In this study, we show that a newly developed ERβ agonist, OSU-ERb-12, can not only impede ovarian cancer cell expansion and tumor growth but also reduce the cancer stem cell (CSC) population. OSU-ERb-12 could inhibit epithelial-to-mesenchymal transition (EMT) in ovarian cancer cells by increasing Snail expression, thereby, blocking EMT-mediated cancer cell dedifferentiation. We also found that OSU-ERb-12 inhibits ovarian cancer expansion in an ERα-independent manner while limiting the CSC population in an ERα-dependent manner. Taken together, our data suggest that the newly developed ERβ agonist OSU-ERb-12 can be used not only to hinder tumor growth but also has the potential to prevent tumor relapse and metastasis by depleting CSCs. Epithelial ovarian cancer is the most lethal malignancy of the female reproductive tract. A healthy ovary expresses both Estrogen Receptor α (ERα) and β (ERβ). Given that ERα is generally considered to promote cell survival and proliferation, thereby, enhancing tumor growth, while ERβ shows a protective effect against the development and progression of tumors, the activation of ERβ by its agonists could be therapeutically beneficial for ovarian cancer. Here, we demonstrate that the activation of ERβ using a newly developed ERβ agonist, OSU-ERb-12, can impede ovarian cancer cell expansion and tumor growth in an ERα-independent manner. More interestingly, we found that OSU-ERb-12 also reduces the cancer stem cell (CSC) population in ovarian cancer by compromising non-CSC-to-CSC conversion. Mechanistically, we revealed that OSU-ERb-12 decreased the expression of Snail, a master regulator of the epithelial-to-mesenchymal transition (EMT), which is associated with de novo CSC generation. Given that ERα can mediate EMT and facilitate maintenance of the CSC subpopulation and that OSU-ERb-12 can block the transactivity of ERα, we conclude that OSU-ERb-12 reduces the CSC subpopulation by inhibiting EMT in an ERα-dependent manner. Taken together, our data indicate that the ERβ agonist OSU-ERb-12 could be used to hinder tumor progression and limit the CSC subpopulation with the potential to prevent tumor relapse and metastasis in patients with ovarian cancer. [ABSTRACT FROM AUTHOR]

Background: Cholangiocarcinoma (CCA) is reported as an aggressive cancer which leads to high mortality and no effective therapeutic target has yet been discovered. Surgical resection is the main method to treat patients with CCA. However, only one-third of CCA patients have the opportunity to accept the operation, leading to poor prognosis for CCA patients. Therefore, it is necessary to search for new therapeutic targets of CCA or core genes involved in the happening and growth of CCA. Aim: In this study, we utilized bioinformatics technology and accessed to several medical databases trying to find the core genes of CCA for the purpose of intervening CCA through figuring out an effective curative target. Methods: Firstly, three differentially expressed genes (DEGs) were discovered from GEPIA, and by further observing the distribution and gene expression, CHST4 was obtained as the core gene. Afterwards, correlated genes of CHST4 in CCA were identified using UALCAN to construct a gene expression profile. We obtained PPI network by Search Tool for the Retrieval of Interacting Networks Genes (STRING) and screened core genes using cytoscape software. Functional enrichment analyses were carried out and the expression of CHST in human tissues and tumors was observed. Finally, a CCA model was established for qPCR and staining validation. Results: Three differentially expressed genes (DEGs), CHST4, MBOAT4 and RP11-525K10.3, were obtained. All were more over-expressed in CCA samples than the normal, among which the change multiple and the gene expression difference of CHST4 was the most obvious. Therefore, CHST4 was selected as the core gene. We can see in our established protein–protein interaction (PPI) network that CHST4 had the highest degree of connectivity, demonstrating its close association with CCA. We found that genes were mainly enriched in CCs in the PPI networks genes which shows functional enrichment analysis results, including golgi lumen, extracellular space and extracellular region. CHST4 was found very specifically expressed in the bile duct and was significantly different from that in normal tissues. The overexpression of CHST4 was further verified in the established animal model of TAA-induced CCA in rats. Quantitative PCR (qPCR) demonstrated that CHST4 was significantly overexpressed in tumor tissues, verifying the role of CHST4 as the core gene of CCA. Conclusion: CHST4 was increasingly expressed in CCA and CHST4 is worth being studied much further in the intervention of CCA. [ABSTRACT FROM AUTHOR]

BACKGROUND: Intestinal amino acid (AA) chemosensing has been implicated in the regulation of AA absorption, nitrogen metabolism and hormone release, thereby playing an indispensable role in maintaining metabolic homeostasis in mammals. The objective of this experiment was to study the distribution of free AA and the expression of AA transporting related genes along the small and large intestines of Liuyang black goats, together with the effects of dietary corn grain replaced by dietary corn gluten feed (CGF). RESULTS: The CGF replacement did not alter (P > 0.05) AA profiles and the expression of AA transporting related genes in the intestinal mucosa. Intriguingly, in terms of gut regions, the concentrations of aspartic acid and glutamic acid in the mucosa of ileum were remarkably less (P < 0.001) than those in the large intestine. Moreover, the concentrations of most cationic and neutral AAs shared the same distribution pattern, with the jejunum and ileum holding the greatest and least levels (P < 0.05), respectively. It was notable that the expression of both anionic and cationic AA transporters in the small intestine was exceedingly greater (P < 0.001) than those in the large intestine. As for transporters of neutral AA, system ASC, L, and A showed an extremely distinctive expression pattern. CONCLUSION: The jejunum would be the primary site of transporting AA, while CGF substitution does not exert a disadvantageous influence on the AA chemosensing systems of the first‐pass metabolism. © 2021 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Tuberculosis caused by the pathogen Mycobacterium tuberculosis (MTB), remains a significant threat to global health. Elucidating the mechanisms of essential MTB genes provides an important theoretical basis for drug exploitation. Gene mtsp17 is essential and is conserved in the Mycobacterium genus. Although Mtsp17 has a structure closely resembling typical steroidogenic acute regulatory protein-related lipid transfer (START) family proteins, its biological function is different. This study characterizes the transcriptomes of Mycobacterium smegmatis to explore the consequences of mtsp17 downregulation on gene expression. Suppression of the mtsp17 gene resulted in significant down-regulation of 3% and upregulation of 1% of all protein-coding genes. Expression of desA1, an essential gene involved in mycolic acid synthesis, and the anti-SigF antagonist MSMEG_0586 were down-regulated in the conditional Mtsp17 knockout mutant and up-regulated in the Mtsp17 over-expression strain. Trends in the changes of 70 of the 79 differentially expressed genes (Log2 fold change > 1.5) in the conditional Mtsp17 knockout strain were the same as in the SigF knockout strain. Our data suggest that Mtsp17 is likely an activator of desA1 and Mtsp17 regulates the SigF regulon by SigF regulatory pathways through the anti-SigF antagonist MSMEG_0586. Our findings indicate the role of Mtsp17 may be in transcriptional regulation, provide new insights into the molecular mechanisms of START family proteins, and uncover a new node in the regulatory network of mycobacteria. [ABSTRACT FROM AUTHOR]

Purpose: Nonalcoholic fatty liver disease (NAFLD) is currently the leading cause of chronic liver disease in developing countries. The pathogenesis is complex, and there is currently no effective treatment. Betaine is an essential intermediate in choline catabolism and an important component of the methionine cycle. Betaine deficiency is associated with NAFLD severity, and its mechanism needs to be further elaborated. Methods: In this study, an NAFLD mouse model was established by feeding ApoE−/− mice a high-fat diet. The effects of betaine on NAFLD were investigated, including its mechanism. Results: In this study, after treatment with betaine, blood lipid levels and liver damage were significantly decreased in the NAFLD mouse model. The fat infiltration of the liver tissues of high-fat diet (HFD)-fed mice after betaine administration was significantly improved. Betaine treatment significantly upregulated AMP-activated protein kinase (AMPK), fibroblast growth factor 10 (FGF10), and adipose triglyceride lipase (ATGL) protein levels both in vivo and in vitro and suppressed lipid metabolism-related genes. Furthermore, the overexpression of FGF10 increased the protein level of AMPK and decreased lipid accumulation in HepG2 cells. Conclusion: Taken together, the data strongly suggest that betaine significantly prevents high-fat diet-induced NAFLD through the FGF10/AMPK signaling pathway in ApoE−/− mice. [ABSTRACT FROM AUTHOR]

Choroidal neovascularization (CNV) is a leading cause of blindness in the elderly in developed countries and is particularly associated with age-related macular degeneration (AMD). Cabozantinib (CBZ) hinders the activation of multiple receptor tyrosine kinases involved in tumor angiogenesis, such as hepatocyte growth factor receptor (MET) and vascular endothelial growth factor receptor 2 (VEGFR2). We aimed to investigate the role and mechanism of CBZ in a mouse laser-induced CNV model. In zebrafish embryos, CBZ perturbed intersegmental vessel (ISV) formation without obvious neurodevelopment impairment. In the mouse laser-induced CNV model, phosphorylated hepatocyte growth factor receptor (p-MET) and phosphorylated vascular endothelial growth factor receptor 2 (p-VEGFR2) were increased in the CNV region. CBZ intravitreal injection or oral gavage alleviated CNV leakage and the CNV lesion area without obvious intraocular toxicity, as well as disturbed the phosphorylation of MET and VEGFR2. Additionally, CBZ downregulated the expression of the hepatocyte growth factor (HGF) with no effect on the expression of the vascular endothelial growth factor (VEGF). CBZ downregulated HGF, p-MET, and p-VEGFR2 expressions in vitro, as well as inhibited the proliferation, migration, and tube formation of b-End3 cells. In summary, CBZ alleviates mouse CNV formation possibly via inhibiting the activation of MET and VEGFR2. The findings provide a novel potential therapy method for CNV patients. [ABSTRACT FROM AUTHOR]

Objectives: Ischemia–reperfusion (I/R) injury can aggravate the dysfunction and structural damage of tissues and organs. This study aimed at investigating the pathogenesis of I/R injury. Methods: GSE82146 was extracted from Gene Expression Omnibus database, which included 12 nonischemic control (NIC) hippocampal tissues and 15 complete global brain ischemia (CGBI)–reperfusion hippocampal tissues. After processing the original data using the affy package, the differentially expressed genes (DEGs) between CGBI and NIC samples were analysed by the limma package. An enrichment analysis for the DEGs was implemented based on the MATHT online tool. Using Cytoscape software, a protein–protein interaction (PPI) network was built and significant network modules were obtained. Finally, miRNA-gene pairs were predicted using the miRWalk2.0 tool, and the miRNA-gene regulatory network was built using the Cytoscape software. Results: Overall, 322 DEGs (279 upregulated and 43 downregulated) were present in the CGBI samples. In PPI network, JUN, STAT3, ATF3, VEGFA and ATF4 had higher degrees. Four significant modules (modules a, b, c and d) were obtained from PPI network. Enrichment analysis suggested that FGF2 in module d was involved in MAPK signalling pathway. In the miRNA-gene regulatory network, rno-miR-125a-5p and rno-miR-125b-5p were among the top 10 miRNAs. Conclusion:JUN, STAT3, ATF3, VEGFA, ATF4, FGF2, rno-miR-125a-5p and rno-miR-125b-5p might affect the development and progression of I/R injury. [ABSTRACT FROM AUTHOR]

Rapamycin (Rap) and its derivatives, called rapalogs, are being explored in clinical trials targeting cancer and neurodegeneration. The underlying mechanisms of Rap actions, however, are not well understood. Mechanistic target of rapamycin (mTOR), a lysosome-localized protein kinase that acts as a critical regulator of cellular growth, is believed to mediate most Rap actions. Here, we identified mucolipin 1 (transient receptor potential channel mucolipin 1 [TRPML1], also known as MCOLN1), the principle Ca2+ release channel in the lysosome, as another direct target of Rap. Patch-clamping of isolated lysosomal membranes showed that micromolar concentrations of Rap and some rapalogs activated lysosomal TRPML1 directly and specifically. Pharmacological inhibition or genetic inactivation of mTOR failed to mimic the Rap effect. In vitro binding assays revealed that Rap bound directly to purified TRPML1 proteins with a micromolar affinity. In both healthy and disease human fibroblasts, Rap and rapalogs induced autophagic flux via nuclear translocation of transcription factor EB (TFEB). However, such effects were abolished in TRPML1-deficient cells or by TRPML1 inhibitors. Hence, Rap and rapalogs promote autophagy via a TRPML1-dependent mechanism. Given the demonstrated roles of TRPML1 and TFEB in cellular clearance, we propose that lysosomal TRPML1 may contribute a significant portion to the in vivo neuroprotective and anti-aging effects of Rap via an augmentation of autophagy and lysosomal biogenesis. [ABSTRACT FROM AUTHOR]

Autophagy is important for cellular survival during renal ischemia/reperfusion (I/R) injury. Ischemic preconditioning (IPC) has a strong renoprotective effect during renal I/R. Our study here aimed to explore the effect of IPC on autophagy during renal I/R injury. Rats were subjected to unilateral renal ischemia with or without prior IPC. Hypoxia/reoxygenation (H/R) injury was induced in HK-2 cells with or without prior hypoxic preconditioning (HPC). Autophagy and apoptosis were detected after reperfusion or reoxygenation for different time. The results showed that the levels of LC3II, Beclin-1, SQSTM1/p62, and cleaved caspase-3 were altered in a time-dependent manner during renal I/R. IPC further induced autophagy as indicated by increased levels of LC3II and Beclin-1, decreased level of SQSTM1/p62, and accumulation of autophagosomes compared to I/R groups at corresponding reperfusion time. In addition, IPC reduced the expression of cleaved caspase-3 and alleviated renal cell injury, as evaluated by the levels of serum creatinine (Scr), neutrophil gelatinase-associated lipocalin (NGAL), and kidney injury molecule-1 (KIM-1) in renal tissues. In conclusion, autophagy and apoptosis are dynamically altered during renal I/R. IPC protects against renal I/R injury and upregulates autophagic flux, thus increasing the possibility for a novel therapy to alleviate I/R-induced acute kidney injury (AKI). [ABSTRACT FROM AUTHOR]

Background: Osteosarcoma (OSA) is the most common bone tumor in children and dogs; however, no substantial improvement in clinical outcome has occurred in either species over the past 30 years. MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and play a fundamental role in cancer. The purpose of this study was to investigate the potential contribution of miR-34a loss to the biology of canine OSA, a well-established spontaneous model of the human disease. Methodology and principal findings: RT-qPCR demonstrated that miR-34a expression levels were significantly reduced in primary canine OSA tumors and canine OSA cell lines as compared to normal canine osteoblasts. In canine OSA cell lines stably transduced with empty vector or pre-miR-34a lentiviral constructs, overexpression of miR-34a inhibited cellular invasion and migration but had no effect on cell proliferation or cell cycle distribution. Transcriptional profiling of canine OSA8 cells possessing enforced miR-34a expression demonstrated dysregulation of numerous genes, including significant down-regulation of multiple putative targets of miR-34a. Moreover, gene ontology analysis of down-regulated miR-34a target genes showed enrichment of several biological processes related to cell invasion and motility. Lastly, we validated changes in miR-34a putative target gene expression, including decreased expression of KLF4, SEM3A, and VEGFA transcripts in canine OSA cells overexpressing miR-34a and identified KLF4 and VEGFA as direct target genes of miR-34a. Concordant with these data, primary canine OSA tumor tissues demonstrated increased expression levels of putative miR-34a target genes. Conclusions: These data demonstrate that miR-34a contributes to invasion and migration in canine OSA cells and suggest that loss of miR-34a may promote a pattern of gene expression contributing to the metastatic phenotype in canine OSA. [ABSTRACT FROM AUTHOR]

Quantitative real-time polymerase chain reaction (qRT-PCR) is one of the most common methods for gene expression studies. Data normalization based on reference genes is essential for obtaining reliable results for qRT-PCR assays. This study evaluated potential reference genes of Chinese yam (Dioscorea opposita Thunb.), which is an important tuber crop and medicinal plant in East Asia. The expression of ten candidate reference genes across 20 samples from different organs and development stages was assessed. We identified the most stable genes for qRT-PCR studies using combined samples from different organs. Our results also suggest that different suitable reference genes or combinations of reference genes for normalization should be applied according to different organs and developmental stages. To validate the suitability of the reference genes, we evaluated the relative expression of PE2.1 and PE53, which are two genes that may be associated with microtuber formation. Our results provide the foundation for reference gene(s) selection in D. opposita and will contribute toward more accurate gene analysis studies of the genus Dioscorea. [ABSTRACT FROM AUTHOR]

Traditional Chinese medicine Jinfukang (JFK) has been clinically used for treating lung cancer. To examine whether epigenetic modifications are involved in its anticancer activity, we performed a global profiling analysis of H3K4Me3, an epigenomic marker associated with active gene expression, in JFK-treated lung cancer cells. We identified 11,670 genes with significantly altered status of H3K4Me3 modification following JFK treatment (P<0.05). Gene Ontology analysis indicates that these genes are involved in tumor-related pathways, including pathway in cancer, basal cell carcinoma, apoptosis, induction of programmed cell death, regulation of transcription (DNA-templated), intracellular signal transduction, and regulation of peptidase activity. In particular, we found that the levels of H3K4Me3 at the promoters of SUSD2, CCND2, BCL2A1, and TMEM158 are significantly altered in A549, NCI-H1975, NCI-H1650, and NCI-H2228 cells, when treated with JFK. Collectively, these findings provide the first evidence that the anticancer activity of JFK involves modulation of histone modification at many cancer-related gene loci. [ABSTRACT FROM AUTHOR]

It has become clear that repetitive sequences have played multiple roles in eukaryotic genome evolution including increasing genetic diversity through mutation, changes in gene expression and facilitating generation of novel genes. However, identification of repetitive elements can be difficult in the ab initio manner. Currently, some classical ab initio tools of finding repeats have already presented and compared. The completeness and accuracy of detecting repeats of them are little pool. To this end, we proposed a new ab initio repeat finding tool, named HashRepeatFinder, which is based on hash index and word counting. Furthermore, we assessed the performances of HashRepeatFinder with other two famous tools, such as RepeatScout and Repeatfinder, in human genome data hg19. The results indicated the following three conclusions: (1) The completeness of HashRepeatFinder is the best one among these three compared tools in almost all chromosomes, especially in chr9 (8 times of RepeatScout, 10 times of Repeatfinder); (2) in terms of detecting large repeats, HashRepeatFinder also performed best in all chromosomes, especially in chr3 (24 times of RepeatScout and 250 times of Repeatfinder) and chr19 (12 times of RepeatScout and 60 times of Repeatfinder); (3) in terms of accuracy, HashRepeatFinder can merge the abundant repeats with high accuracy. [ABSTRACT FROM AUTHOR]

Genes in the signal transducer and activator of transcription (STAT) family are vital for activities including gene expression and immune response. To investigate the functions of the silkworm Bombyx mori STAT ( Bm-STAT ) gene in antiviral immunity, two Bm-STAT gene isoforms, Bm-STAT -L for long form and Bm-STAT- S for short form, were cloned. Sequencing showed that the open reading frames were 2313 bp encoding 770 amino acid residues for Bm-STAT -L and 2202 bp encoding 734 amino acid residues for Bm-STAT- S. The C-terminal 42 amino acid residues of Bm-STAT-L were different from the last 7 amino acid residues of Bm-STAT-S. Immunofluorescence showed that Bm-STAT was primarily distributed in the nucleus. Transcription levels of Bm-STAT in different tissues were determined by quantitative PCR, and the results revealed Bm-STAT was mainly expressed in testes. Western blots showed two bands with molecular weights of 70 kDa and 130 kDa in testes, but no bands were detected in ovaries by using anti-Bm-STAT antibody as the primary antibody. Expression of Bm-STAT in hemolymph at 48 h post infection with B. mori macula-like virus (BmMLV) was slightly enhanced compared with controls, suggesting a weak response induced by infection with BmMLV. Hemocyte immunofluorescence showed that Bm-STAT expression was elevated in B. mori nucleopolyhedrovirus (BmNPV)-infected cells. Moreover, resistance of BmN cells to BmNPV was reduced by downregulation of Bm-STAT expression and increased by upregulation. Resistance of BmN cells to BmCPV was not significantly improved by upregulating Bm-STAT expression. Therefore, we concluded that Bm-STAT is a newly identified insect gene of the STAT family. The JAK-STAT pathway has a more specialized role in antiviral defense in silkworms, but JAK-STAT pathway is not triggered in response to all viruses. [ABSTRACT FROM AUTHOR]

We used samples from rheumatoid arthritis (RA) patients to examine whether Anti-citrullinated protein antibodies (ACPAs) alter macrophage subset distribution and promote RA development. Macrophage subset distributions and interferon regulatory factor 4 (IRF4) and IRF5 expressions were analyzed. ACPAs were purified by affinity column. After RA and osteoarthritis (OA) patients' macrophages were cocultured with ACPAs, macrophage subsets and IRF4 and IRF5 expressions were measured. Small interfering RNAs (siRNAs) were transfected into ACPA-activated cells to suppress IRF4 or IRF5. Fluorescence-activated cell sorting (FACS), Western blot, and immunohistochemistry were performed. Macrophage subset disequilibrium occurred in RA patient synovial fluids. IRF4 and IRF5 were all expressed in the synovial fluid and synovium. ACPAs (40 IU/ml) could induce macrophages to polarize to M1 subsets, and the percentage of increased M1/M2 ratio of RA patients was higher than that of the OA patients. ACPAs also induce IRF4 and IRF5 protein expressions. IRF5 siRNA transfection impaired ACPA activity significantly. We demonstrated that macrophage subset disequilibrium occurred in RA patients. ACPAs induced IRF5 activity and led to M1 macrophage polarization. [ABSTRACT FROM AUTHOR]

The expression pattern of Sohlh1 (spermatogenesis and oogenesis specific basic helix-loop-helix 1) and Sohlh2 in mice has been reported in previous studies. Sohlh1 and Sohlh2 are specifically expressed in spermatogonia, prespermatogonia in male mice and oocytes of primordial and primary follicles in female mice. In this report, we studied the expression pattern of Sohlh1 and Sohlh2 in human adult tissues. Immunohistochemical staining of Sohlh1 and Sohlh2 was performed in 5 samples of normal ovaries and testes, respectively. The results revealed that Sohlh genes are not only expressed in oocytes and spermatogonia, but also in granular cells, theca cells, Sertoli cells and Leydig cells, and in smooth muscles of blood vessel walls. To further investigate the expression of Sohlh genes in other adult human tissues, we collected representative normal adult tissues developed from three embryonic germ layers. Compared with the expression in mice, Sohlhs exhibited a much more extensive expression pattern in human tissues. Sohlhs were detected in testis, ovary and epithelia developed from embryonic endoderm, ectoderm and tissues developed from embryonic mesoderm. Sohlh signals were found in spermatogonia, Sertoli cells and also Leydig cells in testis, while in ovary, the expression was mainly in oocytes of primordial and primary follicles, granular cells and theca cells of secondary follicles. Compared with Sohlh2, the expression of Sohlh1 was stronger and more extensive. Our study explored the expression of Sohlh genes in human tissues and might provide insights for functional studies of Sohlh genes. [ABSTRACT FROM AUTHOR]

Background: Angiotensin II (AngII) participates in endothelial damage and inflammation, and accelerates atherosclerosis. Endothelial lipase (EL) is involved in the metabolism and clearance of high density lipoproteins (HDL), the serum levels of which correlate negatively with the onset of cardiovascular diseases including atherosclerosis. However, the relationship between AngII and EL is not yet fully understood. In this study, we investigated the effects of AngII on the expression of EL and the signaling pathways that mediate its effects in human umbilical vein endothelial cells (HUVECs). Methods and Findings: HUVECs were cultured in vitro with different treatments as follows: 1) The control group without any treatment; 2) AngII treatment for 0 h, 4 h, 8 h, 12 h and 24 h; 3) NF-κB activation inhibitor pyrrolidine dithiocarbamate (PDTC) pretreatment for 1 h before AngII treatment; and 4) mitogen-activated protein kinase (MAPK) p38 inhibitor (SB203580) pretreatment for 1 h before AngII treatment. EL levels in each group were detected by immunocytochemical staining and western blotting. HUVECs proliferation was detected by MTT and proliferating cell nuclear antigen (PCNA) immunofluorescence staining. NF-kappa B (NF-κB) p65, MAPK p38, c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK) and phosphorylated extracellular signal-regulated kinase (p-ERK) expression levels were assayed by western blotting. The results showed that the protein levels of EL, NF-κB p65, MAPK p38, JNK, and p-ERK protein levels, in addition to the proliferation of HUVECs, were increased by AngII. Both the NF-kB inhibitor (PDTC) and the MAPK p38 inhibitor (SB203580) partially inhibited the effects of AngII on EL expression. Conclusion: AngII may upregulate EL protein expression via the NF-κB and MAPK signaling pathways. [ABSTRACT FROM AUTHOR]

Highlights: [•] LPS reduces the expression of GDNF in Sertoli cells and its conditioned media but not BMP4 or SCF. [•] LPS can decrease the expression of SSC self-renewal genes and increase the expression of SSC differentiation genes. [•] LPS can also depress the expression of GDNF downstream genes. [ABSTRACT FROM AUTHOR]

Context: Our previous study demonstrated that tetrandrine (TET) could reverse the resistance of Candida albicans to fluconazole. Objective: The aim of this study was to investigate the molecular mechanism underlying this action. Materials and methods: Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) was performed to compare the expression levels of the drug resistance genes CDR1, CDR2, MDR1, FLU1 and ERG11 in fluconazole-sensitive CA-3 and resistant CA-16 cells that were either treated with FLC and/or TET or left as untreated controls. In addition, intracellular ATP levels were measured using an ATP assay kit, and the expression level of the energy metabolism gene ADH1 was measured by real-time RT-PCR. Results: Compared with FLC/TET-free conditions, FLC + TET treatment strains showed statistically different ( p < 0.05) expression of CDR1 and CDR2 (increased in the FLC-sensitive strains, while decreased in the FLC-resistant strains), MDR1 (increased in the FLC-resistant strains), FLU1 and ERG11 (increased in the FLC-sensitive strains), ADH1 (decreased in both the FLC-sensitive and the FLC-resistant strains). And also, the FLC + TET treatment decreased the intracellular ATP levels in both the FLC-sensitive and the FLC-resistant strains ( p < 0.05). Discussion and conclusion: These results suggest that changes in the expression levels of the drug resistance genes CDR1 and CDR2, the cellular ATP supply and the expression level of the energy metabolism gene ADH1 contribute to the TET-mediated reversal of the fluconazole resistance of C. albicans. [ABSTRACT FROM AUTHOR]

Abstract: In recent years, the potential value of the non-structural proteins (NSP) 2C and 3ABC has been well documented for differentiation of animals infected with foot-and-mouth disease virus (FMDV) from vaccinated animals (DIVA). In order to develop a more sensitive approach to detect animals infected naturally in herds of FMDV-vaccinated animals, a 47.6kD fusion protein named 2C3AB was expressed in bacteria which incorporated two major B-cell epitope regions of 2C and the whole 3AB within the NSP of FMDV. The product reacted specifically with sera from animals infected with FMDV, but did not react with sera from non-vaccinated and healthy animals. The performance of 2C3AB was compared further with the 3ABC fusion protein as the antigen in an indirect ELISA format for DIVA. The results showed that the 2C3AB-ELISA had an even stronger signal reaction in the indirect ELISA and showed higher sensitivity than the 3ABC-ELISA for DIVA purposes and for detection of early virus infection in animals. Therefore, it is expected that the recombinant protein 2C3AB could be a good candidate protein with which to develop more sensitive methods for DIVA and for surveillance of herds infected subclinically under conditions of vaccination. This study indicates that the 2C3AB-ELISA can be used to confirm the results of the 3ABC-ELISA to improve the performance of the 3ABC-ELISA DIVA test. [Copyright &y& Elsevier]

Abstract: Rett syndrome is a neurodevelopmental disorder caused by Mecp2 gene mutations. In RTT patients and Mecp2-null (Mecp2 −/Y) mice, norepinephrine (NE) content drops significantly, which may play a role in breathing arrhythmia, sleep disorders and sudden death. However, the underlying mechanisms for the NE defect are not fully understood. The NE defect may result from decreased NE biosynthesis, loss of catecholaminergic neurons or both. Although deficiency in tyrosine hydroxylase (TH) has been demonstrated, it is possible that dopamine β-hydroxylase (DBH), the critical enzyme converting dopamine to NE, is also affected. To test these possibilities, we studied DBH expressions in pontine catecholaminergic neurons of Mecp2 −/Y mice identified with breathing abnormalities. In comparison to the wild type, Mecp2 −/Y mice at 2months of age showed ∼50% decrease in the expressions of DBH and TH, at both protein and mRNA levels in the locus coeruleus (LC) region. Consistently, DBH and TH immunoreactivity was markedly decreased in LC neurons of Mecp2 −/Y mice. No evidence was found for selective deficiency in TH- or DBH-containing neurons in Mecp2 −/Y mice, as almost all TH-positive cells expressed DBH. By counting TH-immunoreactive cells in the LC, we found that the Mecp2 −/Y mice lost only ∼5% of the catecholaminergic neurons as compared to wild-type, although their LC volume shrank by ∼15%. These results strongly suggest that the NE defect in Mecp2 −/Y mice is likely to result from deficient expression of not only TH but also DBH without significant loss of catecholaminergic neurons in the LC. [Copyright &y& Elsevier]

Opisthorchis felineus , Opisthorchis viverrini , and Clonorchis sinensis (family Opisthorchiidae) are parasitic flatworms that pose serious threats to humans in certain countries and cause opisthorchiasis/clonorchiasis. Opisthorchiid flukes parasitize the biliary tract of the host, causing cholangitis, cholecystitis, cholelithiasis and cholangiocarcinoma. In this review, we primarily focus on recent microRNAs (miRNAs) studies of opisthorchiid flukes and their definitive hosts. Many miRNAs are conserved and expressed in a developmentally stage specific manner in the three opisthorchiid flukes, which play important roles in the growth and development of Opisthorchiidae spp., as well as host-pathogen interactions. Some miRNAs might be potential biomarkers related to carcinogenesis of cholangiocarcinoma. Therefore, this review provides the basis for further investigating the roles of miRNAs in opisthorchiid flukes and their definitive hosts, as well as promoting the development of novel approaches to prevent and treat opisthorchiasis/clonorchiasis. • MicroRNAs are widely expressed in opisthorchiid flukes and their defintive hosts. • The expression pattern of miRNAs is species-specific and dependent on the developmental stage. • A total of 55 conserved miRNAs, belonging to 34 families, were shared by O. felineus, O. viverrini and C. sinensis. • The top three expressed miRNAs in adults of all three species were miR-10-P2a, miR-71-P1, and miR-281. • The MiRNA bantam may help regulate cell proliferation and miR-192 might be a biomarker of O. viverrini -associated carcinogenesis [ABSTRACT FROM AUTHOR]

AbstractBreast cancer is a heterogeneous disease that is ranked as one of the most common cancers worldwide. Currently, although there are existing molecules such as progesterone receptor and estrogen receptor for breast cancer treatment, discovering more effective drug targets is still in urgent need. In this study, we have obtained six sequencing datasets of breast cancer from GEO database and identified a set of differentially expressed molecules, including 67 miRNAs and 133 genes. Function enrichment analysis by miRPathDB database indicated that targets of 11 miRNAs could be enriched in breast cancer pathway with a p-value ≤ .05. A special miRNA-gene interaction network was constructed for analysis of the progression of breast cancer. We then ranked the importance of each molecule (i.e. miRNA and gene) by their node centrality indexes in the network and selected the top 10% of molecules. The statistical analysis of these molecules showed three miRNAs (hsa-miR-1275, hsa-miR-2392, hsa-miR-3141) have significant effects on the prognosis and survival of patients. By searching for potential drugs in Drugbank database, we have identified four candidates (phenethyl isothiocyanate, amuvatinib, theophylline, trifluridine) for targeting these genes. In conclusion, we believe that these drugs and their analogs could be used in the targeted therapy of breast cancer in the future. [ABSTRACT FROM AUTHOR]

Class-3 semaphorins (SEMA3s), initially characterized as axon guidance cues, have been recognized as key regulators for immune responses, angiogenesis, tumorigenesis and drug responses. The functions of SEMA3s are attributed to the activation of downstream signaling cascades mainly mediated by cell surface receptors neuropilins (NRPs) and plexins (PLXNs), yet their roles in human cancers are not completely understood. Here, we provided a detailed pan-cancer analysis of NRPs and PLXNs in their expression, and association with key signal transducers, patient survival, tumor microenvironment (TME), and drug responses. The expression of NRPs and PLXNs were dysregulated in many cancer types, and the majority of them were further dysregulated in metastatic tumors, indicating a role in metastatic progression. Importantly, the expression of these genes was frequently associated with key transducers, patient survival, TME, and drug responses; however, the direction of the association varied for the particular gene queried and the specific cancer type/subtype tested. Specifically, NRP1, NRP2, PLXNA1, PLXNA3, PLXNB3, PLXNC1, and PLXND1 were primarily associated with aggressive phenotypes, whereas the rest were more associated with favorable prognosis. These data highlighted the need to study each as a separate entity in a cancer type- and subtype-dependent manner. [ABSTRACT FROM AUTHOR]

• RPG supplementation attenuated local inflammation and promoted epithelial metabolism in the ileum. • RPG supplementation did not alter ileal microbiota in both digesta and mucosa. • RPG can serve as an effective dietary intervention to improve productive performance and health status of cows. Optimizing diet formula with the aim of alleviating negative energy balance (NEB) is of great significance to the production performance of transition dairy cows. The objective of the study was to evaluate the effects of rumen-protected glucose (RPG) supplementation on ileal microbiota, the expression of genes involved in epithelial metabolism and immune homeostasis in the ileum of transition dairy cows. Ten Holstein dairy cows (age, 4.7 ± 0.5 years; bodyweight, 515 ± 42 kg; parity, 2.8 ± 0.4; body condition score, 3.2 ± 0.3; milk yield, 16.1 ± 3.7 kg/d) were randomly divided into two groups. The treatments were a basal diet plus 200 g/d of RPG (450 g/kg of glucose, 450 g/kg of coating fat and 100 g/kg of water) or a basal diet plus 90 g/d of coating fat (CON). Cows were provided with the diets for 21 days (from 1 week prepartum to 2 weeks postpartum) and euthanized on d 22 for taking samples. The ileal digesta samples were taken from the mid-ileum to analyze volatile fatty acid concentration and composition of digesta-associated microbiota. The ileal mucosa samples were taken from the mid-ileum for analysis of gene expression and composition of mucosa-associated microbiota. Ileal total volatile fatty acid concentration was lower (P = 0.018) in the PRG-supplemented group than in the CON group. Compared with CON group, the PRG supplementation up-regulated or tended to up-regulate (P < 0.10) the expression of PC , SGLT3, IGFBP5, PPARA, IL10 , Occludin , and TPJ1 , but down-regulated the expression of IL17A (P = 0.032) and tended to down-regulate (P < 0.10) the expression of TLR4 , TLR9 and IL26. However, no differences (P > 0.10) were observed between the RPG and CON groups in both ileal total bacterial copy number and ileal bacterial beta diversity. Collectively, dietary RPG supplementation promoted epithelial metabolism and improved immune homeostasis in the ileum, with no influence on both digesta- and mucosa-associated ileal microbiota of transition dairy cows. [ABSTRACT FROM AUTHOR]

Osteosarcoma (OS) is a leading malignant tumor reported with high mortality and morbidity. Dysexpression of CircBBS9 has been reported to exhibit a critical functional role in various diseases. However, the underlying molecular mechanisms of CircBBS9 in osteosarcoma are poorly characterized. The present study aims to investigate the impacts of CircBBS9 on the progression of osteosarcoma. The findings of the study demonstrated the up-regulated expression of CircBBS9 in osteosarcoma. The Actinomycin D and RNase R treatment experiments confirmed that circBBS9 is indeed a circRNA. In addition, the knockdown of circBBS9 negatively impacted the migration, proliferation and invasion of osteosarcoma cells. Further investigations illustrated that circBBS9 controlled miR-485-3p and miR-485-3p might directly interact with HMGB1. miR-485-3p had a negative regulatory role in HMGB1's gene expression. Through rescue assays, it was verified that CircBBS9 promoted osteosarcoma progression through the miR-485-3p/HMGB1 axis. Finally, circBBS9 knockdown attenuated the in-vivo growth of osteosarcoma. Conclusively, our study is the first time to examine the possible functional mechanism and regulation roles of CircBBS9 in osteosarcoma. The findings explained that CircBBS9 promoted the malignant osteosarcoma's progression by sponging miR-485-3p/HMGB1 and proposed CircBBS9 as a prognostic biomarker and therapeutic candidate for osteosarcoma patients. [ABSTRACT FROM AUTHOR]

Pancreatic cancer, mostly pancreatic ductal adenocarcinoma (PDAC), is one of the most lethal cancer types, with an estimated 44,330 death in 2018 in the US alone. While targeted therapies and immune checkpoint inhibitors have significantly improved treatment options for patients with lung cancer and renal cell carcinomas, little progress has been made in pancreatic cancer, with a dismal 5-year survival rate currently at ~8%. Upon diagnosis, the majority of pancreatic cancer cases (~80%) are already metastatic. Thus, identifying ways to reduce pancreatic cancer metastasis is an unmet medical need. Furthermore, pancreatic cancer is notorious resistant to chemotherapy. While Kirsten RAt Sarcoma virus oncogene (K-RAS) mutation is the major driver for pancreatic cancer, specific inhibition of RAS signaling has been very challenging, and combination therapy is thought to be promising. In this study, we report that combination of hedgehog (Hh) and Mitogen-activated Protein/Extracellular Signal-regulated Kinase Kinase (MEK) signaling inhibitors reduces pancreatic cancer metastasis in mouse models. In mouse models of pancreatic cancer metastasis using human pancreatic cancer cells, we found that Hh target gene Gli1 is up-regulated during pancreatic cancer metastasis. Specific inhibition of smoothened signaling significantly altered the gene expression profile of the tumor microenvironment but had no significant effects on cancer metastasis. By combining Hh signaling inhibitor BMS833923 with RAS downstream MEK signaling inhibitor AZD6244, we observed reduced number of metastatic nodules in several mouse models for pancreatic cancer metastasis. These two inhibitors also decreased cell proliferation significantly and reduced CD45+ cells (particularly Ly6G+CD11b+ cells). We demonstrated that depleting Ly6G+ CD11b+ cells is sufficient to reduce cancer cell proliferation and the number of metastatic nodules. In vitro, Ly6G+ CD11b+ cells can stimulate cancer cell proliferation, and this effect is sensitive to MEK and Hh inhibition. Our studies may help design novel therapeutic strategies to mitigate pancreatic cancer metastasis. [ABSTRACT FROM AUTHOR]

Background Marijuana consumption is on the rise in the US but the health benefits of cannabis smoking are controversial and the impact of cannabis components on lung homeostasis is not well-understood. Lung function requires a fine regulation of the ion channel CFTR, which is responsible for fluid homeostasis and mucocilliary clearance. The goal of this study was to assess the effect that exposure to Δ9-tetrahydrocannabinol (THC), the psychoactive substance present in marijuana, has on CFTR expression and function. Methods Cultures of human bronchial epithelial cell line 16HBE14o- and primary human airway epithelial cells were exposed to THC. The expression of CFTR protein was determined by immunoblotting and CFTR function was measured using Ussing chambers. We also used specific pharmacological inhibitors of EGFR and ERK to determine the role of this pathway in THC-induced regulation of CFTR. Results THC decreased CFTR protein expression in primary human bronchial epithelial cells. This decrease was associated with reduced CFTR-mediated short-circuit currents. THC also induced activation of the ERK MAPK pathway via activation of EGFR. Inhibition of EGFR or MEK/ERK prevented THC-induced down regulation of CFTR protein expression. Conclusions and general significance THC negatively regulates CFTR and this is mediated through the EGFR/ERK axis. This study provides the first evidence that THC present in marijuana reduces the expression and function of CFTR in airway epithelial cells. [ABSTRACT FROM AUTHOR]

Radish (Raphanus sativus L.) is an important cool-season root vegetable crop worldwide. Heat stress (HS) severely restricts radish taproot formation and diminishes its yield and quality. Although WRKY transcription factors (TFs) play pivotal roles in several biotic and/or abiotic stress responses, how WRKY TFs mediate HS regulatory network remains elusive in radish. Herein, a WRKY-Ia TF gene, RsWRKY26 , exhibited dramatically up-regulated expression under heat stress, especially at 8 h and 24 h. RsWRKY26 was localized to the nucleus and displayed transactivation activity in both the yeast and plant cells. Interestingly, the RsWRKY26- overexpressing radish plants exhibited significantly enhanced thermotolerance compared to the control plants, whereas its knock-down plants showed increased thermosensitivity. Both in vitro and in vivo assays showed that RsWRKY26 activated RsHSP70–20 transcription by directly binding to the specific W-box element in its promoter. Moreover, several biochemical assays revealed that a VQ motif-containing protein RsVQ4 interacted with RsWRKY26, which promoted DNA-binding activity of RsWRKY26 to regulate RsHSP70–20 expression. Notably, RsWRKY26 in turn binds to RsVQ4 promoter to activate its transcription, thereby forming a positive feedback loop to cooperatively regulate heat stress response in radish. Together, we revealed a novel RsVQ4-RsWRKY26- RsHSP70–20 module to promote the thermotolerance, which would facilitate dissecting the molecular regulatory network of RsWRKY-mediated thermal morphogenesis in radish. • RsWRKY26 positively regulates thermotolerance by activating RsHSP70-20 transcription. • RsVQ4 physically interacts with RsWRKY26 to promote its DNA-binding activity. • RsWRKY26 binds to RsVQ4 promoter to form a feedback loop enhancing thermotolerance. [ABSTRACT FROM AUTHOR]