Alternative splicing refers to the process in which different mRNA splice isomers are produced from an mRNA precursor by different splicing methods. WRKY transcription factors are one of the largest transcription factor families and are widely involved in growth, development, aging and abiotic stress processes. In this study, we found that WRKY28-2 from Caragana korshinskii can undergo alternative splicing and produce two different transcripts, CkWRKY28-2 and CkWRKY28-2-AS. The start codon of CkWRKY28-2 is ATG, the end codon is TGA, encoding 265 aa, the molecular weight of CkWRKY28-2 protein is 30 028.18 Da, and the pI is 8.16. The start codon of CkWRKY28-2-AS is ATG, the stop codon is TGA, encoding 68 aa, the molecular weight of CkWRKY28-2-AS is 7 286.71 Da, and the pI is 3.98. The results of RT-qPCR analysis showed that CkWRKY28-2 was mainly expressed in roots and stems, while CkWRKY28-2-AS was mainly expressed in leaves. Under drought stress, CkWRKY28-2 expression increased while CkWRKY28-2-AS expression decreased. Under salt stress, the expression levels of CkWRKY28-2 and CkWRKY28-2-AS increased with increasing salt treatment time. Under high temperature stress, the expression levels of CkWRKY28-2 and CkWRKY28-2-AS were significantly down-regulated compared with the control. Under low temperature stress, the expression levels of CkWRKY28-2 and CkWRKY28-2-AS decreased significantly with increasing treatment time. This study laid a foundation for the molecular mechanism of CkWRKY28-2 involved in stress resistance.
Objective: When cAMP was synthesized via the salvage pathway, guanosine was also produced in large amounts as a major by-product. In order to enhance cAMP salvage synthesis, an inosine monophosphate dehydrogenase (IMPDH) inhibitor was screened and utilized to reduce by-product production. Methods: The optimum addition conditions for IMPDH were determined by shake flask experiments and the fermentations with oxymatrine addition were carried out in a 7 L fermentor. Then, the fermentation performance, key enzyme activities, by-product levels and transcriptome analysis were systematically investigated. Results: With the addition of 20 mg/L oxymatrine for 24 h, the cAMP concentration reached 4.30 g/L with an increase of 58.67% higher than that of the control, and the fermentation performance was also obviously promoted. The results of enzyme activities and by-product levels showed that IMPDH together with 5'-nucleotidase were strongly inhibited and the enzyme activities for cAMP synthesis were enhanced, obviously leading to higher cAMP levels and lower guanosine production. The transcriptome analysis results showed that due to the addition of oxymatrine, the transcription levels of key enzyme genes in EMP, de novo synthesis and by-product synthesis pathway were significantly decreased, and at the same time, those in TCA cycle, PPP pathway and salvage pathway were significantly increased. Conclusion: Oxymatrine could effectively decrease by-product synthesis and promote cAMP synthesis by regulating metabolism of enzyme activity and transcription level, resulting in more carbon flow and ATP supply directed into cAMP savage pathway for product synthesis.
Various microorganisms in nature provide a vast library of genetic elements for heavy metal bioadsorption. The analysis of the mechanism of microbial heavy metal resistance and the study of related genes are prerequisites for the use of these genetic elements. In this study, we screened five highly mercury-tolerant strains from mercury-contaminated soils in mining areas. The genetic relationship of each strain was investigated by 16S rRNA sequencing analysis. The mercury tolerance of the strains was further tested. HG5, the closest relative of Peribacillus frigoritolerans, was the more mercury tolerant strain, still able to grow colonies in the medium containing 150 mg/L HgCl2. Transcriptome analysis of HG5 under mercury stress showed that nucleotide synthesis and metabolism, heme synthesis and metabolism, pyruvate metabolism, TCA cycle and other pathways were affected by mercury. At the same time, quantitative real-time PCR proved that mercury stress had a major impact on the sulfur assimilation process of HG5. After mercury stress, the expression of the trehalose operon gene increased significantly. This study could provide a theoretical basis for the utilization of heavy metal resistance genes and related gene elements.
Objective: To use bioinformatics methods to screen and experimentally validate long non-coding RNA (lncRNA) associated with oxidative stress (OS) in hepatocellular carcinoma (HCC) in relation to prognosis. Methods: We first downloaded HCC-related expression profile data and clinical data from The Cancer Genome Atlas (TCGA) database, and analyzed differentially expressed genes (DEGs) and OS-related lncRNA using R language. We then verified the reliability of the risk model using receiver operating characteristic (ROC) curves and LASSO regression analysis. Kaplan-Meier survival analysis was used to evaluate patient survival. We used Nomogram to construct a model of pathologic variables and risk scores for predicting 1-year, 3-year, and 5-year survival rates of patients. The bioinformatics results were validated in vitro by RT-qPCR, colony formation, cell scratch, Transwell and cell cycle. Results: Eight prognostic lncRNAs (TMEM220-AS1, AC012360.2, SNHG3, GASAL1, PCAT6, AC009005.1, AL031985.3, AC009403.1) were acquired through regression analysis and were utilized to establish the prognostic model. AC009005.1 was highly expressed in tumor cells, and interference with AC009005.1 significantly inhibited the invasion, migration and proliferation of tumor cells.
Exosomes, nanoscale extracellular vesicles secreted by cells, play a critical role in mediating intercellular communication and transporting a variety of bioactive substances, including nucleic acids, proteins, and lipids. These vesicles play a significant role in both physiological and pathological processes of the body. Due to their endogenous origin and biocompatibility, exosomes offer distinct advantages over synthetic carriers such as liposomes and nanoparticles, particularly in the area of disease diagnostics, biomarker discovery, and therapeutic applications. However, their clinical utility is currently hampered by challenges such as low efficiency in isolation and purification, and limited targeting capabilities. This article provides a comprehensive overview of exosomes, including their origin, classification, and recent advances in isolation and purification techniques. It critically evaluates the strengths and limitations of these methods. Additionally, the review discusses the burgeoning applications of exosomes in disease diagnostics and therapeutics, highlighting their potential and future prospects. By providing novel insights and strategies, this review aims to pave the way for more effective use of exosomes in clinical and research settings, thereby unlocking their full potential for healthcare innovation.
Currently, one of the most promising tools in tumor immunotherapy is dendritic cell (DC) vaccines. DC vaccines are based on the uptake of antigens delivered by DCs and the activation of T cells to induce anti-tumor effects. However, traditional DC vaccines have made slow progress in the treatment of solid tumors due to the limited activity of monocyte-derived DCs and tumor-mediated immunosuppression. Novel DC vaccines and DC vaccine combination therapies offer new ways to improve tumor treatment compared to traditional DC vaccine therapy. This article reviews the major challenges faced by traditional DC vaccines in the field of solid tumor therapy, the current research progress of novel DC vaccines, and DC vaccine combination therapy strategies. It also discusses their development prospects and provides new ideas for future research and product development of DC vaccines in tumor therapy.
Cancer remains one of the leading causes of human mortality. Despite significant advances in cancer treatment by modern medicine, traditional therapeutic approaches still face limitations, including inadequate targeting and significant toxic side effects. Nevertheless, the emergence of technological advancements has brought a ray of hope in tumor treatment through the utilization of bacteria and their derivatives, which have unique biological properties. Bacteria, due to their inherent survival characteristics, exhibit highly effective tumor-targeting colonization and direct tumor cell killing capabilities while being amenable to genetic engineering. Consequently, through genetic modification, bacteria and their derivatives can be used as efficient drug delivery vehicles, seamlessly integrating various treatment modalities, including radiotherapy, chemotherapy, and immunotherapy. The combination aims to overcome therapeutic deficiencies, increase the efficacy of tumor treatment, and minimize toxic side effects. This review comprehensively summarizes the latest research progress in optimizing combination therapy strategies using bacteria and their derivatives for tumor treatment. Specifically, it explores the advantages of utilizing bacteria and their derivatives as drug carriers in tumor treatment and outlines methods for engineering bacteria for drug delivery, and elucidates strategies for optimizing combination therapy. The primary objective is to provide a theoretical basis for optimizing anti-tumor treatment strategies, ultimately unlocking the therapeutic potential of bacteria, enhancing the effectiveness of anti-tumor treatment, and providing safer and more effective treatment options for people fighting cancer.
As a serious liver disease, the clinical treatment of liver failure faces significant challenges. Chronic liver disease, viral hepatitis infection, drug poisoning, and other factors can lead to liver failure, threatening the lives of thousands of patients. In recent years, stem cell therapy has emerged as a novel treatment method, offering a new approach to the treatment of liver failure. It works by promoting the repair and regeneration of liver cells, regulating the immune response, and inhibiting liver fibrosis and inflammation. Several clinical trials have demonstrated that stem cell transplantation therapy can significantly improve the survival rate of patients with liver failure. It also effectively improves liver function and blood clotting function. However, further studies are needed to confirm its long-term effectiveness and safety. This paper summarizes the research progress of stem cell therapy for liver failure, discusses its potential therapeutic mechanism, and explores its prospects for clinical application. The aim is to provide a reference for future research and clinical practice.
The human intestinal microbiota is closely related to the onset and development of human diseases, and it has become an important clinical treatment tool to regulate the structure and function of the intestinal flora for the treatment of diseases. Plant essential oil have become a hotspot of current research due to their advantages of multifaceted regulation of intestinal flora and low toxic side effects. Taking the role of plant volatile oils on intestinal flora and the association between intestinal microbiota and various types of diseases as a starting point, this paper reviews the research on the intrinsic mechanism of plant volatile oils regulating intestinal flora to exert pharmacological effects at home and abroad in recent years to provide a new way of thinking for the research on the pharmacological effects of volatile oils.
Inhalation drug delivery is an emerging approach to disease treatment that offers unique advantages over oral and intravenous routes. However, there are several physiological barriers that affect delivery efficacy. Therefore, research has focused on optimizing carrier design, with inhalable nanoparticles demonstrating excellent design advantages and functional integration capabilities during the delivery process. In this review, we outline inhalation drug delivery devices and formulations and discuss the challenges of inhalation delivery. In particular, we highlight inhalation delivery strategies based on various nanoparticles over the past five years and provide perspectives on the development of inhalation drug delivery nano-platforms, with the aim of providing insights for further research and innovation in the field of inhalation drug delivery.
Terpenoids are widely used in medicine, food, fragrances, and flavors. However, natural terpenoids are produced at low concentrations in plants, making extraction costly and sources limited. The chemical synthesis of terpenoids, especially those with complex chemical structures, is challenging due to harsh conditions and low yields, hindering their widespread application. Biosynthesis offers a promising alternative for efficient production of terpenoids and has become a focal point of research in recent years. With advances in synthetic biology, the construction of microbial cell factories for terpenoid biosynthesis through microbial host modification, novel pathway design, and integration of microbial mass fermentation shows great potential for the food, medical, and cosmetic industries. Terpenoid flavors, which have distinctive aromas, and latent fragrance compounds, which release aromatic components when transformed, are the most economically valuable substances in the industrialization of natural products. This article reviews the synthetic mechanisms, recent advances, and engineering strategies for the biosynthesis of terpenoid flavors and latent fragrances. Key strategies include gene editing, transcriptional regulation, modular pathway engineering, cell membrane engineering, adaptive laboratory evolution, and co-culture systems. The goal is to advance terpenoid synthesis technologies for industrial applications in the fragrance and flavor industry.
