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Background: Neuronal abnormalities are closely associated with major depressive disorder (MDD). Available evidence suggests a role for microRNAs (miRNAs) in regulating the expression of genes involved in MDD. Hence, miRNAs that can be potential therapeutic targets need to be identified. Methods: A mouse model of chronic unpredictable stress (CUS) was used to evaluate the function of miRNAs in MDD. miR‐144‐5p was screened from the hippocampi of CUS mice based on sequencing results. Adenovirus‐associated vectors were used to overexpress or knockdown miR‐144‐5p in mice. BpV(pic) and LY294002 were used to determine the relationship between miR‐144‐5p target genes PTEN and TLR4 in neuronal impairment caused by miR‐144‐5p deficiency. Western blotting, immunofluorescence, ELISA immunosorbent assay, and Golgi staining were used to detect neuronal abnormalities. Serum samples from healthy individuals and patients with MDD were used to detect miR‐144‐5p levels in the serum and serum exosomes using qRT‐PCR. Results: miR‐144‐5p expression was significantly decreased within the hippocampal dentate gyrus (DG) of CUS mice. Upregulation of miR‐144‐5p in the DG ameliorated depression‐like behavior in CUS mice and attenuated neuronal abnormalities by directly targeting PTEN and TLR4 expression. Furthermore, miR‐144‐5p knockdown in normal mice led to depression‐like behavior via inducing neuronal abnormalities, including abnormal neurogenesis, neuronal apoptosis, altered synaptic plasticity, and neuroinflammation. miR‐144‐5p deficiency‐mediated neuronal impairment was mediated by PI3K/Akt/FoxO1 signaling. Furthermore, miR‐144‐5p levels were downregulated in the sera of patients with MDD and associated with depressive symptoms. Consistently, serum exosome‐derived miR‐144‐5p levels were decreased in patients with MDD. Conclusion: miR‐144‐5p plays a vital role in regulating neuronal abnormalities in depression. Our findings provide translational evidence that miR‐144‐5p is a new potential therapeutic target for MDD. [ABSTRACT FROM AUTHOR]

The β-ketoacyl CoA synthase encoded by the KCS genes is a rate-limiting enzyme for the synthesis of very-long-chain fatty acid (VLCFA), which catalyzes the VLCFA elongation. Yellow horn (Xanthoceras sorbifolium) is a horticultural tree species known for its kernel oil, which has strong resistance to drought, cold, high temperature, and saline-alkali. The conserved domain FAE1-CUT1-RPPA and ACP-syn-III_C of the KCS gene family were used to search the KCS sequences across the whole genomic sequence of yellow horn; a total of 20 XsKCS genes were identified and divided into four subfamilies. The conserved motif and transmembrane structure analysis revealed that most XsKCSs had a conserved transmembrane domain except XsKCS10 and XsKCS20. The prediction of cis-acting elements of XsKCS genes showed that XsKCS genes contained many stress and hormone response elements, such as ABRE, MBS, and LTR. Furthermore, XsKCS genes exhibited differential expression profiles under abiotic stress and stress-related hormone treatment conditions. Transcriptomic data showed that XsKCS1, XsKCS11, and XsKCS17 had higher expression in yellow horn with high leaf cuticular wax, indicating that they may function in the cuticular wax accumulation and drought response. This study laid a foundation for further functional verification of XsKCS genes in yellow horn stress response. [ABSTRACT FROM AUTHOR]

Chemical nitrogen (N) fertilizer is widely used in sugarcane production, especially in China and India. Understanding the molecular mechanisms and mining miRNAs and their target genes associated with nitrogen use efficiency (NUE) in sugarcane can aid in developing the N-efficient varieties, and thus is beneficial to reduce N fertilizer application. In this study, the root miRNA database of N-efficient sugarcane variety ROC22 under low N stress (0.3 mM NH4NO3) for 3 h was constructed, along with their transcriptome-rearranged data. KEGG analysis indicated that those candidate target genes, corresponding to differentially expressed miRNAs, were mainly enriched in N metabolism, amino acid metabolism, carbohydrate metabolism, photosynthesis, and hormone signal transduction pathways. It was found that under low N stress for 0–24 h, there was a negative correlation between miR168 and SPX, along with miR396 and acnA. Furthermore, the expression of miR156 in the roots of ROC22 was significantly up-regulated under low N treatment. Compared with the wild-type, the Arabidopsis plants overexpressing sugarcane miR156 exhibited significantly improved length and surface area of roots, while the expression of one NO3− transporter gene NRT1.1, three N assimilation key genes (NR1, NIR1, and GS), and the activity of two N assimilation key enzymes (NR and GS) were up-regulated under low N treatment. It can be reasonably deduced that sugarcane miR156 can enhance the nitrogen assimilation ability of the overexpressed Arabidopsis plants under low N application, and thus has a potential ability for improving sugarcane NUE. The present study should be helpful for understanding the molecular regulatory network in the N-efficient sugarcane genotype responding to low N stress and could provide the candidate miRNAs with a potential function in improving sugarcane NUE. [ABSTRACT FROM AUTHOR]

The investigation into viable but non-culturable (VBNC) bacteria through the implementation of resuscitation promoting factors (Rpfs) has broadened the potential sources for isolating strains capable of degrading polychlorinated biphenyls (PCBs). Nonetheless, there has been limited research on the efficacy of resuscitated strains and the potential improvement of their performance through co-cultivation. In this work, the PCB degradation potential of resuscitated strains, specifically Pseudomonas sp. HR1 and Achromobacter sp. HR2, as well as their co-cultures, was investigated. Of particular importance was the comparative analysis between the optimal co-culture and individual strains regarding their ability to degrade PCB homologs and mineralize intermediate metabolites. The results suggested that the resuscitated strains HR1 and HR2 demonstrated robust growth and effective degradation of Aroclor 1242. The co-culture CO13, with an optimal HR1 to HR2 ratio of 1:3, exhibited a remarkable improvement in PCB degradation and intermediate metabolite mineralization compared to individual strains. Analysis of functional genes and degradation metabolites revealed that both the individual strains and co-culture CO13 degraded PCBs via the HOPDA-benzoate pathway, then mineralized through protocatechuate meta - and ortho -cleavage pathways, as well as the catechol ortho -cleavage pathway. This study represents the first documentation of the improved PCB degradation through the co-cultivation of resuscitated strains, which highlights the great promise of these resuscitated strains and their co-cultures as effective bio-inoculants for enhanced bioremediation. [Display omitted] • First report on co-culture of resuscitated Pseudomonas and Achromobacter strains. • Resuscitated strains HR1 and HR2 exhibited distinct PCB degradation advantages. • CO13 consisting of HR1 and HR2 in 1:3 ratio achieved highest PCB degradation. • Co-culture CO13 surpassed single strains in intermediate metabolite mineralization. [ABSTRACT FROM AUTHOR]

Background. Idiopathic pulmonary fibrosis (IPF) is a progressive disease with high mortality and poor prognosis. The prognostic signatures related to conventional therapy response remain limited. The Wenfei Buqi Tongluo (WBT) formula, a traditional Chinese medicine (TCM) formula, has been widely utilized to treat respiratory diseases in China, which is particularly effective in promoting inflammatory absorption. In this study, we aim to explore the mechanism of the WBT formula in the inhibition of inflammatory response during IPF, based on network pharmacology and in vivo experiments. Methods. Network pharmacology was applied to predict the changes of biological processes and potential pathways for the WBT formula against IPF. Histopathological changes, inflammatory factors (IL-6, IL-1β, and TNF-α), and the proteins of the TLR4/MyD88/NF-κB pathway in bleomycin- (BLM-) induced mice model were examined by hematoxylin-eosin (H&E), Masson or immunohistochemistry staining, Western blot, and enzyme-linked immunosorbent assay analysis. Results. A total of 163 possible components and 167 potential targets between the WBT formula and IPF were obtained. The enrichments of network pharmacology showed that inflammation response, TNF, and NF-κB pathways were involved in the treatment of WBT against IPF. The in vivo experiments indicated that the WBT formula could ameliorate inflammatory exudation and collagen deposition at a histopathology level in the BLM-induced mice model. The levels of IL-6, IL-1β, and TNF-α were reduced after the WBT formula treatment. Moreover, the expressions of phosphorylated-NF-κB p65, TLR4, and MyD88 were significantly downregulated by the WBT formula, compared with the BLM-induced group. Conclusion. These results indicated that the WBT formula can suppress BLM-induced IPF in a mouse model by inhibiting the inflammation via the TLR4/MyD88/NF-κB pathway. This study provides a new insight into the molecular mechanisms of the WBT formula in the application at the clinic. [ABSTRACT FROM AUTHOR]

The exploration of resuscitated strains, facilitated by the resuscitation promoting factor (Rpf), has substantially expanded the pool of cultivated degraders, enhancing the screening of bio-inoculants for bioremediation applications. However, it remains unknown whether these resuscitated strains can re-enter the viable but non-culturable (VBNC) state and the specific stress conditions that trigger such a transition. In this work, the whole genome, and polychlorinated biphenyl (PCB)-degrading capabilities of a resuscitated strain HR1, were investigated. Notably, the focus of this exploration was on elucidating whether HR1 would undergo a transition into the VBNC state when exposed to low temperature and PCBs, with and without the presence of heavy metals (HMs). The results suggested that the resuscitated strain Pseudomonas sp. HR1 harbored various functional genes related to xenobiotic biodegradation, demonstrating remarkable efficiency in Aroclor 1242 degradation and strong resistance against stress induced by low temperature and PCBs. Nevertheless, when exposed to the combined stress of low temperature, PCBs, and HMs, HR1 underwent a transition into the VBNC state. This transition was characterized by significant decreases in enzyme activities and notable changes in both morphological and physiological traits when compared to normal cells. Gene expression analysis revealed molecular shifts underlying the VBNC state, with down-regulated genes showed differential expression across multiple pathways and functions, including oxidative phosphorylation, glycolysis, tricarboxylic acid cycle, amino acid metabolism, translation and cytoplasm, while up-regulated genes predominantly associated with transcription regulation, membrane function, quorum sensing, and transporter activity. These findings highlighted the great potential of resuscitated strains as bio-inoculants in bioaugmentation and shed light on the survival mechanisms of functional strains under stressful conditions, which should be carefully considered during bioremediation processes. [Display omitted] • First report on stress survival strategy of resuscitated strain Pseudomonas sp. HR1. • HR1 can efficiently degrade PCBs and resist stresses from low temperature and PCBs. • HR1 entered VBNC state with joint stress of low temperature, PCBs, and heavy metals. • The VBNC cells of HR1 showed significant morphological and physiological changes. • Induction of the VBNC state in HR1 involved extensive regulation of gene expression. [ABSTRACT FROM AUTHOR]

Background. Acute kidney injury (AKI) caused by bee stings is common, with characteristics of acute onset, severe illness, and high mortality. Melittin, a major component of bee venom, has been considered to play a key role in bee sting related AKI. This study aims to illustrate whether melittin could lead to apoptosis of renal tubular epithelial cells (RTECs) and to investigate its mechanism. Methods. In vivo, 45 mice were randomly divided into the melittin group (n=30, injected with melittin into the tail vein according to the total dose of 4.0 ug/g weight) and the control group (n=15, injected with the same volume of saline into the tail vein). In vitro, human RTECs (HK-2) were cultured and treated with melittin (2ug/ml or 4ug/ml) and TNF-α (10ng/ml). Biochemical analysis, HE stains, and electron microscope were performed to evaluate renal function and pathological changes. TUNEL stains and flow cytometry were performed to detect apoptosis. Real-time PCR was performed to detect mRNA levels of Bax, Bcl-2, and TNF-α. Simple western assay and immunohistochemical (IH) and immunofluorescent (IF) stains were performed for protein detection. Results. Melittin successfully induced AKI in mice. Compared with the control group, obvious injury and apoptosis of RTECs were observed in the melittin group; the mRNA and protein expressions of Bax were significantly increased, while the expression of Bcl-2 was significantly decreased. The serum TNF-αlevel in melittin group was significantly higher than that in control group. In vitro, the results confirmed that melittin can cause HK-2 cells apoptosis. The trends of expression of Bax and Bcl-2 were consistent with the results in vivo. The levels of TNF-α mRNA and protein by PCR and Western blot were significantly higher in melittin group than those in control group. Conclusion. Melittin can lead to the apoptosis of RTECs, which may be mediated by upregulating the expression of Bax/Bcl-2 and activating the TNF-α signaling pathway. [ABSTRACT FROM AUTHOR]

We developed sugarcane plants with improved resistance to the sugarcane borer, Diatraea saccharalis (F). An expression vector pGcry1Ac0229, harboring the cry1Ac gene and the selectable marker gene, bar, was constructed. This construct was introduced into the sugarcane cultivar FN15 by particle bombardment. Transformed plantlets were identified after selection with Phosphinothricin (PPT) and Basta. Plantlets were then screened by PCR based on the presence of cry1Ac and 14 cry1Ac positive plantlets were identified. Real-time quantitative PCR (RT-qPCR) revealed that the copy number of cry1Ac gene in the transgenic lines varied from 1 to 148. ELISA analysis showed that Cry1Ac protein levels in 7 transgenic lines ranged from 0.85 μg/FWg to 70.92 μg/FWg in leaves and 0.04 μg/FWg to 7.22 μg/FWg in stems, and negatively correlated to the rate of insect damage that ranged from 36.67% to 13.33%, respectively. Agronomic traits of six transgenic sugarcane lines with medium copy numbers were similar to the non-transgenic parental line. However, phenotype was poor in lines with high or low copy numbers. Compared to the non-transgenic control plants, all transgenic lines with medium copy numbers had relatively equal or lower sucrose yield and significantly improved sugarcane borer resistance, which lowered susceptibility to damage by insects. This suggests that the transgenic sugarcane lines harboring medium copy numbers of the cry1Ac gene may have significantly higher resistance to sugarcane borer but the sugarcane yield in these lines is similar to the non-transgenic control thus making them superior to the control lines. [ABSTRACT FROM AUTHOR]

Accumulating evidence from previous studies suggested that interleukin-1 (IL-1β) and tumor necrosis factor-α (TNF-α) play an important role in pathogenesis of temporomandibular disorders (TMD). However, the cell surface receptors and the intracellular signal pathways leading to these cytokines expression are not fully understood. In the current study, we investigated the roles of Toll-like receptor 4 (TLR4) and its adaptor myeloid differentiation factor 88 (MyD88) in the expression of IL-1β and TNF-α in synovial fibroblasts (SFs) separated from rat temporomandibular joint (TMJ) with lipopolysaccharide (LPS) stimulation. The results showed that treatment with LPS could increase TLR4, MyD88, IL-1β, and TNF-α expression at both mRNA and protein levels. In addition, increased expression of IL-1β and TNF-α could be blocked by treatment with TAK-242, a blocker of TLR4 signaling, and also by MyD88 inhibitory peptide (MIP). These findings suggested that maybe TLR4/MyD88 signal transduction pathway participates in enhanced expression of IL-1 and TNF-α in patients with TMD. The activation of TLR4/MyD88 signal transduction pathway which results in production of proinflammatory factors may play a role in the pathogenesis of TMD. [ABSTRACT FROM AUTHOR]

Heterologous expression is an efficient alternative to conventional extraction to produce a specific Buthus martensii Karsch ( Bm K) peptide. In this work, Bm K1 was successfully expressed in Escherichia coli after genetic codon optimization, but Bm K1 content was <6% of total cellular protein. To improve Bm K1 expression, a trc promoter library with a wide relative strength was constructed, and three promoters, PpJF136 (0.55), PpJF325 (1.29), and PpJF288 (2.31), were selected to control Bm K1 expression. A higher Bm K1 expression (>13.9% of total protein) was obtained using a stronger promoter, PpJF325. Furthermore, a maximum Bm K1 content (>21.7% of total protein) was obtained by combining promoter PpJF325 and three copies of the Bm K1 gene. The yield of the purified Bm K1 achieved 196.74 mg L−1 in E. coli BL21( DE3) p JF431, which was improved 2.09-fold compared with the control. This was the highest reported production of scorpion peptides in E. coli. [ABSTRACT FROM AUTHOR]

Diabetes is a common, complex, chronic metabolic disease. A randomized, double-blind, placebo-parallel controlled clinical study has shown that Gegen Qinlian Decoction (GQD) can reduce glycosylated hemoglobin in type 2 diabetes mellitus (T2DM) intestinal damp-heat syndrome patients in a dose-dependent manner. To explore the pathogenesis of T2DM intestinal damp-heat syndrome and the therapeutic effect of GQD from the perspective of exosomal microRNA (miRNA). Eligible patients were selected and treated with GQD for 3 months to evaluate their clinical efficacy. Effective cases were matched with healthy volunteers, and saliva samples were collected. Exosomal miRNA was extracted from saliva and analyzed by chip sequencing. Subsequently, the function of the differential gene and the signal transduction pathway were analyzed using bioinformatics technology. Finally, three target miRNAs were randomly selected from the T2DM group/healthy group, and two target miRNAs in the T2DM before treatment/after treatment group were randomly selected for qPCR verification. Finally, we conducted a correlation analysis of the miRNAs and clinical indicators. The registration number for this research is ChiCTR-IOR-15006626. (1) The expression of exosomal miRNA chips showed that there were 14 differentially expressed miRNAs in the T2DM group/healthy group, and 26 differentially expressed miRNAs in the T2DM before treatment/after treatment group. (2) Enrichment results showed that in the T2DM group/healthy group, it was primarily related to cell development, body metabolism, TGF-β, and ErbB signaling pathways. In the T2DM before treatment/after treatment group, it was mainly related to cellular metabolic regulation processes, and insulin, Wnt, and AMPK signaling pathways. (3) The qPCR verification showed that the expressions of hsa-miR-9-5p, hsa-miR-150–5p, and hsa-miR-216b-5p in the T2DM group was higher (P<0.05). Following GQD treatment, hsa-miR-342–3p and hsa-miR-221–3p were significantly downregulated (P<0.05). (4) hsa-miR-9-5p was positively correlated with BMI (P<0.05), and hsa-miR-150–5p was positively correlated with total cholesterol and triglycerides (P<0.05). The GQD efficacy-related gene hsa-miR-342–3p was positively correlated with the patient's initial blood glucose level (P<0.05), and hsa-miR-221–3p was positively correlated with total cholesterol and triglycerides (P<0.05). The exosomal miRNA expression profile and signaling pathways related to T2DM intestinal damp-heat syndrome and the efficacy of GQD were established, which provides an alternative strategy for precision traditional Chinese medicine treatment. [Display omitted] [ABSTRACT FROM AUTHOR]

Starch is the major constituent of green banana fruit and is a resource for banana quality as it is degraded to soluble sugars during postharvest ripening. However, the molecular basis of starch degradation in different banana cultivars during fruit ripening remains largely unclear. In this study, the starch degradation of four banana cultivars (Baxi, Dajiao, Fengjiao, Gongjiao) during ripening was investigated. The contents of total starch, amylase, amylopectin, the amylose/amylopectin ratio, and the crystallinity were all decreased in all the four banana cultivars tested. In addition, the morphological structures of starch granules changed as ripening progressed due to starch degradation. The activity and gene expression of starch hydrolytic enzymes were increased. Importantly, a transcriptional regulator MabZIP21 was identified, which functions as an activator of starch degradation by upregulating the transcription of MaBAM4 , MaBAM7 , and MaAMY3. In addition, transient overexpression of MabZIP21 in banana or overexpression of MabZIP21 in tomato increased starch corruption by elevating the accumulation of starch degrading genes. Through this study, we expect to show that distinct patterns of starch degradation exist in different banana cultivars, and that MabZIP21 mediates starch breakdown by transcriptional activation of MaBAM4 , MaBAM7 , and MaAMY3. ● Starch breaks down during the ripening of banana fruit. ● Starch crystallinity and starch granule size decreased during ripening. ● Amylase and isoamylase activities and transcripts increased in ripening bananas. ● MabZIP21 trans-activates the transcription of MaBAM4 , MaBAM7 and MaAMY3. [ABSTRACT FROM AUTHOR]