Plant-derived artificial miRNA effectively reduced the proliferation of aphid (Aphidoidea) through spray-induced gene silencing.

BACKGROUND: Aphids (Hemiptera: Aphididae) are notorious sap-sucking insects that rampantly threaten agricultural production worldwide. Current management against aphids in the field heavily relies on chemical pesticides, which makes economical and eco-friendly methods urgently needed. Spray-induced gene silencing (SIGS) offers a powerful and precise approach to pest management. However, the high costs and instability of double-stranded RNA (dsRNA) regulators applied for downstream RNA interference (RNAi) still limit this strategy. It remains uncertain if RNAi regulators applied in SIGS could extend to small RNA (sRNA), especially miRNA. RESULTS: We chose two sRNA sequences, miR-9b and miR-VgR, whose corresponding targets ABCG4 and VgR are both essential for aphid growth and development. The efficacy of these sequences was initially verified by chemically synthetic single-stranded RNA (syn-ssRNA). Through spray treatment, we observed a significantly decreased survival number and increased abnormality rate of green peach aphids fed on the host under laboratory conditions. Based on our previous study, we generated transgenic plants expressing artificial miR-9b (amiR-9b) and miR-VgR (amiR-VgR). Remarkably, plant-derived amiRNA exerted potent and long-lasting inhibitory efficacy with merely one percent concentration of chemical synthetics. Notably, the simultaneous application of amiR-9b and amiR-VgR exhibited superior inhibitory efficacy. CONCLUSION: We explored the potential use of sRNA-based biopesticide through SIGS while investigating the dosage requirements. To optimize this strategy, the utilization of plant-derived amiRNA was proposed. The results suggested that attributed to stability and durability, deploying amiRNA in pest management is a potential and promising solution for the field application. © 2024 Society of Chemical Industry.

Transcription suppression of GABARAP mediated by lncRNA XIST-EZH2 interaction triggers caspase-11-dependent inflammatory injury in ulcerative colitis.

BACKGROUND: We have previously found that enhancer of zeste homolog 2 (EZH2) is correlated with inflammatory infiltration and mucosal cell injury in ulcerative colitis (UC). This study aims to analyze the role of X-inactive specific transcript (XIST), a possible interactive long non-coding RNA of EZH2, in UC and to explore the mechanisms. METHODS: C57BL/6N mice were treated with dextran sulfate sodium (DSS), and mouse colonic mucosal epithelial cells were treated with DSS and lipopolysaccharide (LPS) for UC modeling. The UC-related symptoms in mice, and the viability and apoptosis of mucosal epithelial cells were determined. Inflammatory injury in animal and cellular models were assessed through the levels of ACS, occludin, IL-1ß, IL-18, TNF-α, caspase-1, and caspase-11. Molecular interactions between XIST, EZH2, and GABA type A receptor-associated protein (GABARAP) were verified by immunoprecipitation assays, and their functions in inflammatory injury were determined by gain- or loss-of-function assays. RESULTS: XIST was highly expressed in DSS-treated mice and in DSS + LPS-treated mucosal epithelial cells. It recruited EZH2, which mediated gene silencing of GABARAP through H3K27me3 modification. Silencing of XIST alleviated body weight loss, colon shortening, and disease active index of mice and reduced inflammatory injuries in their colon tissues. Meanwhile, it reduced apoptosis and inflammation in mucosal epithelial cells. However, these alleviating effects were blocked by either EZH2 overexpression or GABARAP knockdown. Rescue experiments identified caspase-11 as a key effector mediating the inflammatory injury following GABARAP loss. CONCLUSION: This study suggests that the XIST-EZH2 interaction-mediated GABARAP inhibition activates caspase-11-dependent inflammatory injury in UC.

The combined effect of fire and nitrogen addition on biodiversity and herbaceous aboveground productivity in a coastal shrubland.

Introduction: Fire and nitrogen (N) deposition each impact biodiversity and ecosystem productivity. However, the effect of N deposition on ecosystem recovery after fire is still far from understood, especially in coastal wetlands. Methods: We selected a typical coastal shrubland to simulate three N deposition levels (0, 10, and 20 g N m-2 year-1) under two different burned conditions (unburned and burned) in the Yellow River Delta of North China. Soil properties, soil microbial biodiversity, shrub growth parameters, herbaceous biodiversity, and aboveground productivity were determined after experimental treatments for 1 year. Results: We found that fire had a stronger influence on the ecosystem than N addition. One year after the fire, shrub growth had significantly decreased, while soil pH, soil electrical conductivity, herbaceous biodiversity, soil microbial biodiversity, and herbaceous aboveground productivity significantly increased. Conversely, a single year of N addition only slightly increased herbaceous aboveground productivity. The combined effect of fire and N addition was only significant for fungus biodiversity and otherwise had minimal influence. Interestingly, we found that herbaceous aboveground productivity was positively associated with fungal community diversity under unburned conditions but not in burned shrublands. Fire showed a great impact on soil parameters and biodiversity in the coastal wetland ecosystem even after a full year of recovery. Discussion: Fire may also diminish the influence of several belowground factors on herbaceous aboveground productivity, which ultimately reduces recovery and stability. Appropriate N addition may be an effective way to improve the ecosystem productivity in a wetland dominated by shrub species.

CCCTC-Binding Factor Mediates the Transcription of Insulin-Like Growth Factor Binding Protein 5 Through EZH2 in Ulcerative Colitis.

BACKGROUND: Ulcerative colitis (UC) features chronic, non-infectious inflammation of the colon. Insulin-like growth factor binding protein 5 (IGFBP5) has been indicated to be related to various inflammation-related diseases. However, its association with UC remains largely unclear. AIMS: Here, we investigate the role of IGFBP5 in colonic mucosal epithelial cell injury in UC. METHODS: Differentially expressed genes in the colonic tissues of UC mice were screened using the Gene Expression Omnibus database, and IGFBP5 was identified. UC mice were developed using dextran sulfate sodium, and IGFBP5 expression in the colonic tissues of UC mice was confirmed by immunohistochemistry and RT-qPCR. The effects of IGFBP5 in vivo and in vitro were investigated by intraperitoneal injection of adeno-associated virus into UC mice or by transfection with an IGFBP5 overexpression plasmid into lipopolysaccharide-treated colonic mucosal epithelial cells. The mechanisms causing IGFBP5 deletion in UC were predicted by bioinformatics analysis and ChIP-qPCR and verified by rescue experiments. RESULTS: IGFBP5 was reduced in UC. IGFBP5 impaired the NFκB pathway in the colonic tissue of UC mice and ameliorated inflammatory infiltration and colonic mucosal cell injury. IGFBP5 depletion was associated with H3K27me3 modification, which was induced by EZH2. CTCF was responsible for recruiting EZH2 to the promoter region of IGFBP5. CTCF inhibition repressed UC progression by reducing H3K27me3 modification via the discouragement of the enrichment of EZH2 in the IGFBP5 promoter. CONCLUSIONS: CTCF modulates H3K27me3 modification of the IGFBP5 promoter by recruiting EZH2, thereby downregulating IGFBP5 to accentuate colonic mucosal epithelial cell injury in UC mice.

lncRNA EGFEM1P promotes thyroid cancer progression by acting as an miR-369-3p sponge and upregulating TCF4.

Thyroid cancer is the most commonly diagnosed endocrine cancer, with the incidence of 14.42 per 100,000 person-years in 2010-2013. It is important to conduct an in-depth exploration into the molecular mechanisms of thyroid cancer, providing insights into the improvements of therapy. Long non-coding RNAs (lncRNAs) act as oncogenes or tumor suppressors in thyroid cancer by sponging microRNAs (miRNAs), however, the functions of numerous lncRNAs are still unknown. In the present study, via the comprehensive analysis of microarray data derived from papillary thyroid tumors and the RNA sequencing of thyroid tumors from The Cancer Genome Atlas database, EGF like and EMI domain containing 1 (EGFEM1P) expression levels in papillary thyroid tumors and normal adjacent tissues were explored. Reverse transcription-quantitative PCR was used to detect EGFEM1P, microRNA (miR)-369-3p and T cell factor 4 (TCF4) expression levels. Western blotting was used to detect TCF4 protein and cleaved caspase-3/8 expression levels. Cell proliferative ability and apoptosis were assessed using the Cell Counting Kit-8 assay and flow cytometry, respectively. The interactions between EGFEM1P and miR-369-3p, and miR-369-3p and TCF4, were determined using the dual-luciferase reporter assay. The results demonstrated that EGFEM1P was upregulated in papillary thyroid tumors and thyroid cancer cells compared with normal adjacent tissues and human normal thyroid epithelial Nthy-ori 3-1 cell line. In the examined thyroid cancer cells, EGFEM1P was demonstrated to interact with miR-369-3p and decreased miR-369-3p expression levels. Thereafter, TCF4 was determined to be a target gene of miR-369-3p and EGFEM1P promoted TCF4 expression via regulating miR-369-3p expression levels. At last, it was found that EGFEM1P expression promoted rapid cell proliferation and inhibited cell apoptosis in thyroid cancer cells via acting as a miR-369-3p sponge. In conclusion EGFEM1P promoted thyroid cancer progression via acting as a sponge of the miR-369-3p/TCF4 axis.

A conserved asparagine residue in the inner surface of BRI1 superhelix is essential for protein native conformation.

Asparagine-linked glycosylation (ALG, N-glycosylation) is one of the most prevalent protein modifications in eukaryotes and regulates protein folding, trafficking and function. Recently, we reported that the mutation of N154Q significantly led to the ER retention of brassinosteroids insensitive 1 (BRI1), the receptor of brassinosteroids (BRs). However, the mechanism of how the N154 site affects BRI1 structure is still not completely clear. In current study, we found that the removal of N154-glycan with S156A replacement significantly enhanced the ability of bri1 to complement bri1-301 mutant and plasma membrane localization compared with N154Q. In addition, the various mutations on N154 site resulted in bri1 retention in the ER, except for N154D. The 3D modeling suggested that there existed polar contacts around N154 site and the mutations not only destroyed the addition of N-glycan on the site, but also led to the disorder of hydrogen bonds formation. The sequence analysis showed that the N275 shared more similarity with N154 site and the removal of N275-glycan further enhanced the retention of bri1 carrying S156A mutation in the ER. Our results showed that N154 was special and essential for maintaining BRI1 structure and explored the role of those residues and key N-glycans lying in the LRR inner surface on protein conformation.

Clinical observation of octreotide combined with diclofenac sodium in preventing ERCP-related pancreatitis.

OBJECTIVE: To elucidate the efficacy of octreotide in combination with diclofenac sodium in the prevention of ERCP-related pancreatitis, and investigate its impact on patients' serum amylase, white blood cell (WBC) count, adverse effects, hyperamylasemia and hemorheology. METHODS: The prospective study was conducted, in which 124 patients who underwent endoscopic retrograde cholangiopancreatography (ERCP) in our hospital were evenly divided into 2 groups, the observation group (n=62) and the control group (n=62), via a random number table method. The control group was administered diclofenac sodium lidocaine hydrochloride via injection after ERCP, while the observation group was given octreotide acetate on the basis of the control group. The incidence of pancreatitis, serum amylase level, WBC count, incidence of adverse effects and hyperamylasemia, and hemorheology levels were compared between these two groups of patients. RESULTS: The incidence of pancreatitis in the observation group was significantly lower than that in the control group (P<0.05). After treatment, the serum amylase level at 24 h post-surgery in the observation group was notably lower than that in the control group (P<0.001). The WBC count at 2 h and 24 h post-surgery in the observation group were both significantly lower than those in the control group (all P<0.001). The incidence of total adverse reactions in the observation group was remarkably lower than that in the control group (P<0.01). The incidence of hyperamylasemia in the observation group was considerably lower than that in the control group (P<0.001). Twenty-four hours post-surgery, the whole blood viscosity at high shear rate, whole blood viscosity at low shear rate, and plasma viscosity in the observation group were all significantly lower than those in the control group (all P<0.001). CONCLUSION: The combination of octreotide and diclofenac sodium could effectively prevent the occurrence of ERCP-related pancreatitis, which reduced the incidence of hyperamylasemia, decreased the WBC count and serum amylase level, improved the hemorheology, and lowered the incidence of adverse effects in patients after ERCP. Therefore, this therapeutic strategy is worthy of clinical propagation and application.

Plant Size Plays an Important Role in Plant Responses to Low Water Availability and Defoliation in Two Woody Leguminosae Species.

Plant size influences plant responses to combined environmental factors under climate change. However, their roles in plant ecophysiological responses are not fully understood. Two rapidly growing Leguminosae species (Robinia pseudoacacia and Amorpha fruticosa) were used to examine plant responses to combined drought and defoliation treatments (two levels of both treatments). Both 1.5 month-old seedlings and 3 month-old seedlings were grown in a greenhouse, and seedling growth, leaf gas exchanges, stem hydraulics, and concentrations of non-structural carbohydrates were determined after 60 days of treatment. Our results indicated defoliation had no significant effect on plant height, basal diameter, and total biomass whatever plant sizes and species. Under the low water availability treatment, the defoliated seedlings significantly increased by 24% in stem water potential compared with non-defoliated seedlings in large R. pseudoacacia. Compared with the high water availability in large non-defoliated R. pseudoacacia seedlings, the low water availability significantly reduced by 26% in stem starch concentration to maintain the stem soluble sugar concentration stable, but not in small R. pseudoacacia seedlings. We also found a negative correlation between leaf and root soluble sugar concentration under low water availability in A. fruticosa. The results demonstrate defoliation could relieve the effect of low water availability in large seedlings. Large seedlings had more compensatory mechanisms in response to defoliation and drought treatments than small seedlings, thus species with large carbon reserves are more recommended for vegetation restoration under combined drought and defoliation conditions. Future studies with more species are crucial for obtaining more rigorous conclusions.

Biodiversity, environmental context and structural attributes as drivers of aboveground biomass in shrublands at the middle and lower reaches of the Yellow River basin.

Biodiversity-biomass relationships have been debated for decades and remain subject to controversy. Understanding the relationship between biodiversity and biomass will be crucial for soil and water conservation of the whole basin of the Yellow River. The positive effects of biodiversity on aboveground biomass are altered via two fundamental mechanisms-niche complementarity and selection effects-and are modulated by environmental context and community structure in natural communities. Most studies of biodiversity-biomass relationships have focused on grasslands and forests, rather than on shrublands. We combine multiple biotic variables (biodiversity, functional identity and community structural attributes) and environmental context with aboveground biomass across shrubland habitat types (temperate hilly, temperate montane and subtropical montane) at neighbourhood and community spatial scales, to evaluate the effects of these factors on shrubland aboveground biomass, in the Yellow River basin. We found aboveground biomass was influenced primarily by the community-weighted mean plant maximum height, followed by species richness and community-weighted mean specific leaf area. Furthermore, individual plant size inequality, mean annual precipitation and water availability either directly or indirectly influenced aboveground biomass. The biodiversity-aboveground biomass relationship was stronger at the broader spatial scale. Thus, our findings indicate that both niche complementarity and selection effects shape the effects of biodiversity on shrubland aboveground biomass, although selection effects are more important. Moreover, they indicate that water is the most important environmental factor for determining aboveground biomass, and suggest that community structure and spatial scale could influence shrubland aboveground biomass and its response to biodiversity.

Identification of the DNA Replication Regulator MCM Complex Expression and Prognostic Significance in Hepatic Carcinoma.

BACKGROUND: The microliposome maintenance (MCM) complex, MCM2-7, is revealed to be involved in multiple cellular processes and plays a key role in the development and progression of human cancers. However, the MCM complex remains poorly elaborated in hepatic carcinoma (HCC). METHODS: In the study, we found the mRNA and protein level by bioinformatics. We also explored the prognostic value, genetic alteration, interaction network, and functional enrichment of MCM2-7. The MCM expression and correlation among these MCMs in HCC cell lines were identified by western blot. RESULTS: MCM2-7 was significantly increased in HCC tissues compared to normal liver tissues. The high level of MCM2-7 had a positive correlation with poor prognosis. However, MCM2-7 alterations were not correlated with poor OS. MCMs were both increased in HCC cell lines compared to the normal hepatocyte cell line. Furthermore, the positive correlation was found among MCMs in HCC cell lines. CONCLUSIONS: The MCM complex was increased in HCC tissues and cell lines and negatively correlated with prognosis, which might be important biomarkers for HCC.

Clinical Significance of Polo-Like Kinase 4 as a Marker for Advanced Tumor Stage and Dismal Prognosis in Patients With Surgical Gastric Cancer.

OBJECTIVE: This study aimed to investigate the correlation of polo-like kinase 4 with clinicopathological features and survival profiles in patients with gastric cancer. METHODS: This retrospective study was conducted based on the clinical data from 289 eligible patients with gastric cancer who received resection. Polo-like kinase 4 expression in adjacent tissue and tumor tissue was determined by immunohistochemical assay and semiquantified scoring method using immunohistochemical score by staining intensity score multiplying staining density score. Based on the total immunohistochemical score (ranged from 0 to 12), the polo-like kinase 4 expression was classified as low expression (immunohistochemical: 0-3) and high expression (immunohistochemical: 4-12); furthermore, high expression was divided into high+ expression (immunohistochemical: 4-6), high++ expression (immunohistochemical: 7-9), and high+++ expression (immunohistochemical: 10-12). RESULTS: Polo-like kinase 4 expression was elevated in tumor tissue compared with adjacent tissue. Tumor polo-like kinase 4 high expression correlated with increased T stage and Tumor, Node, Metastasis (TNM) stage, while, it did not correlate with age, gender, current smoke, current drink, chronic complications, Helicobacter pylori infection, tumor location, pathological grade, or N stage. Besides, higher tumor polo-like kinase 4 expression correlated with shorter disease-free survival and overall survival. Subsequently, multivariate Cox proportional hazards regression analysis showed that higher tumor polo-like kinase 4 expression was an independent predictive factor for worse disease-free survival but not for overall survival. CONCLUSION: Polo-like kinase 4 possesses the clinical significance as a biomarker for aiding prognostication and facilitating postoperative tumor management in patients with gastric cancer.

Functional identity enhances aboveground productivity of a coastal saline meadow mediated by Tamarix chinensis in Laizhou Bay, China.

Research in recent decades has confirmed that biodiversity influences ecosystem productivity; however, the potential mechanisms regulating this process remain subject to controversy, due to variation across ecosystems. Here, the effects of biodiversity on ecosystem productivity were evaluated using three variables of biodiversity (taxonomic diversity, functional identity, and functional diversity) and surrounding environmental conditions in a coastal saline meadow located on the south coast of Laizhou Bay, China. At this site, the shrub and field layers were primarily dominated by Tamarix chinensis and natural mesic grasses, respectively. Our results showed that functional identity, which is quantified as the community weighted mean of trait values, had greater explanatory ability than taxonomic and functional diversity. Thus, ecosystem productivity was determined disproportionately by the specific traits of dominant species. T. chinensis coverage was a biotic environmental factor that indirectly affected ecosystem productivity by increasing the community weighted mean of plant maximum height, which simultaneously declined with species richness. The present study advances our understanding of the mechanisms driving variation in the productivity of temperate coastal saline meadows, providing evidence supporting the "mass ratio" hypothesis.

Bacterial community shaped by heavy metals and contributing to health risks in cornfields.

Scientists are increasingly aware that heavy metal contamination in soils, especially in farmland ecosystems, can negatively affect human health and alter the bacterial community that plays a critical role in plant growth and heavy metal accumulation. The goal of the present paper was to uncover how various heavy metals and non-metallic elements affect human health and bacterial diversity in cornfields and to explore the contribution of soil bacteria to heavy metal accumulation in crops. Soil samples were collected from five counties in Shandong Province, China, where abnormally high levels of heavy metals and metalloids were caused by mining and heavy industry. We calculated a hazard quotient (HQ) to evaluate the health risk these heavy metals cause and analyzed the soil bacterial community using 16S rRNA gene sequencing. The HQ results showed that As posed the greatest threat to human health followed by Pb although concentrations of all metals did not reach the health risk threshold. Meanwhile, principal component analysis (PCA) and redundancy analysis (RDA) revealed soil bacterial richness was significantly influenced by As, Ni, and Cr as well as pH and phosphorus, but not by the species diversity of aboveground weeds. The most abundant bacteria in our study region were heavy metal tolerant groups, specifically Actinobacteria and Proteobacteria. Moreover, correlation analysis suggested that Actinobacteria might reduce the phytoaccumulation of Cr, Cu, Zn, and Hg in corn, while Proteobacteria might weaken phytoaccumulation of Pb, Ni, As, and Cd. Our results verified that heavy metals play an important role in shaping the soil bacterial community. Using native bacteria in farmland provides a potential biological strategy for reducing the health risk posed by heavy metals related to food consumption.