Animal venoms are a complex mixture of highly specialized toxic molecules. Among them, pore-forming proteins (PFPs) or toxins (PFTs) are one of the major disease-causing toxic elements. The ability of the PFPs in defense and toxicity through pore formation on the host cell surface makes them unique among the toxin proteins. These features made them attractive for academic and research purposes for years in the areas of microbiology as well as structural biology. All the PFPs share a common mechanism of action for the attack of host cells and pore formation in which the selected pore-forming motifs of the host cell membrane-bound protein molecules drive to the lipid bilayer of the cell membrane and eventually produces water-filled pores. But surprisingly their sequence similarity is very poor. Their existence can be seen both in a soluble state and also in transmembrane complexes in the cell membrane. PFPs are prevalent toxic factors that are predominately produced by all kingdoms of life such as virulence bacteria, nematodes, fungi, protozoan parasites, frogs, plants, and also from higher organisms. Nowadays, multiple approaches to applications of PFPs have been conducted by researchers both in basic as well as applied biological research. Although PFPs are very devastating for human health nowadays researchers have been successful in making these toxic proteins into therapeutics through the preparation of immunotoxins. We have discussed the structural, and functional mechanism of action, evolutionary significance through dendrogram, domain organization, and practical applications for various approaches. This review aims to emphasize the PFTs to summarize toxic proteins together for basic knowledge as well as to highlight the current challenges, and literature gap along with the perspective of promising biotechnological applications for their future research.

Gastric cancer (GC) is one of the main causes of cancer-related death. Lysine acetyltransferases 2 A (KAT2A) is a succinyltransferase that plays an essential role in cancer development. The pyruvate kinase M2 (PKM2) is a glycolysis rate-limiting enzyme that mediates the glycolysis of cancers. This study aimed to explore the effects and mechanism of KAT2A in GC progression. The effects of biological behaviors of GC cells were evaluated by MTT, colony formation and seahorse assays. The succinylation modification was assessed by immunoprecipitation (IP). The interaction between proteins were detected by Co-IP and immunofluorescence. A pyruvate kinase activity detection kit was used to evaluate the activity of PKM2. Western blot was performed to detect the expression and oligomerization of protein. Herein, we confirmed that KAT2A was highly expressed in GC tissues and was associated with a poor prognosis. Function studies showed that knockdown of KAT2A inhibited cell proliferation and glycolytic metabolism of GC. Mechanistically, KAT2A could directly interacted with PKM2 and KAT2A silencing inhibited the succinylation of PKM2 at K475 site. In addition, the succinylation of PKM2 altered its activity rather than its protein levels. Rescue experiments showed that KAT2A promoted GC cell growth, glycolysis, and tumor growth by promoting PKM2 K475 succinylation. Taken together, KAT2A promotes the succinylation of PKM2 at K475 to inhibit PKM2 activity, thus promotes the progression of GC. Therefore, targeting KATA2 and PKM2 may provide novel strategies for the treatment of GC.

MicroRNAs play a key role in the pathogenesis of many types of cancer, including thyroid cancer (TC). MiR-138-5p has been confirmed to be abnormally expressed in TC tissues. However, the role of miR-138-5p in TC progression and its potential molecular mechanism need to be further explored. In this study, quantitative real-time PCR was used to examine miR-138-5p and TRPC5 expression, and western blot analysis was performed to examine the protein levels of TRPC5, stemness-related markers, and Wnt pathway-related markers. Dual-luciferase reporter assay was used to assess the interaction between miR-138-5p and TRPC5. Cell proliferation, stemness, and apoptosis were examined using colony formation assay, sphere formation assay, and flow cytometry. Our data showed that miR-138-5p could target TRPC5 and its expression was negatively correlated with TRPC5 expression in TC tumor tissues. MiR-138-5p decreased proliferation, stemness, and promoted gemcitabine-induced apoptosis in TC cells, and this effect could be reversed by TRPC5 overexpression. Moreover, TRPC5 overexpression abolished the inhibitory effect of miR-138-5p on the activity of Wnt/β-catenin pathway. In conclusion, our data showed that miR-138-5p suppressed TC cell growth and stemness via the regulation of TRPC5/Wnt/β-catenin pathway, which provided some guidance for studying the potential function of miR-138-5p in TC progression.

A lot of research has been done on using natural items as diabetes treatment. The molecular docking study was conducted to evaluate the inhibitory activities of urolithin A against α-amylase, α-glucosidase, and aldose reductase. The molecular docking calculations indicated the probable interactions and the characteristics of these contacts at an atomic level. The results of the docking calculations showed the docking score of urolithin A against α-amylase was -5.169 kcal/mol. This value for α-glucosidase and aldose reductase was -3.657 kcal/mol and -7.635 kcal/mol, respectively. In general, the outcomes of the docking calculations revealed that urolithin A can construct several hydrogen bonds and hydrophobic contacts with the assessed enzymes and reduces their activities considerably. The properties of urolithin against common human breast cancer cell lines, i.e., SkBr3, MDA-MB-231, MCF-7, Hs578T, Evsa-T, BT-549, AU565 and 600MPE were evaluated. The IC50 of the urolithin was 400, 443, 392, 418, 397, 530, 566 and 551 against SkBr3, MDA-MB-231, MCF-7, Hs578T, Evsa-T, BT-549, AU565 and 600MPE, respectively. After doing the clinical trial studies, the recent molecule may be used as an anti-breast cancer supplement in humans. IC50 values of urolithin A on α-amylase, α-glucosidase, and aldose reductase enzymes were obtained at 16.14, 1.06 and 98.73 µM, respectively. A lot of research has been done on using natural items as diabetes treatment. The molecular docking study was conducted to evaluate the inhibitory activities of urolithin A against alpha-amylase, alpha-glucosidase, and aldose reductase. The properties of urolithin against common human breast cancer cell lines, i.e., SkBr3, MDA-MB-231, MCF-7, Hs578T, Evsa-T, BT-549, AU565 and 600MPE were evaluated. After doing the clinical trial studies, the recent molecule may be used as an anti-breast cancer supplement in humans. IC50 values of urolithin A on alpha-amylase, alpha-glucosidase, and aldose reductase enzymes were obtained at 16.14, 1.06 and 98.73 µM, respectively.

Disturbance of extravillous trophoblast infiltration is associated with preeclampsia (PE), a severe condition of pregnancy characterized by hypertension and proteinuria. Senescence-associated epithelial membrane protein 1 (SEMP1), an integral membrane protein, is a vital component of tight junction strands in epithelial or endothelial cells, with no clear function reported in PE. Gene Expression Omnibus (GEO) datasets showed that SEMP1 expression was downregulated in the placental tissues of PE patients, which was confirmed by assessing SEMP1 levels in placental samples collected in our hospital. Furthermore, less SEMP1 was detected in cytokeratin 7 positive trophoblast cells in the spiral arteries of rat placentas post L-arginine methyl ester hydrochloride (L-NAME) treatment. Trophoblast cells acquired robust ability of proliferation, migration, and invasion when SEMP1 was overexpressed. Such capability was weakened in SEMP1-silenced cells. Trophoblast cells overexpressing SEMP1 secreted more vascular endothelial growth factor A (VEGFA), which facilitated the tube formation of human umbilical vein endothelial cells. Blockade of PI3K/AKT signaling transduction with LY294002 dampened the effects of SEMP1 on trophoblast cells. Collectively, we firstly indicated that SEMP1 inhibition is a potential driver for PE, which may be associated with the deactivation of the PI3K/AKT pathway. SEMP1 was involved in PE progression by regulating cell growth, migration, invasion, and tube formation via PI3K/AKT signaling pathway in trophoblast cells and endothelial cells.

The CRISPR/Cas system, an innovative gene-editing tool, is emerging as a promising technique for genome modifications. This straightforward technique was created based on the prokaryotic adaptive immune defense mechanism and employed in the studies on human diseases that proved enormous therapeutic potential. A genetically unique patient mutation in the process of gene therapy can be corrected by the CRISPR method to treat diseases that traditional methods were unable to cure. However, introduction of CRISPR/Cas9 into the clinic will be challenging because we still need to improve the technology's effectiveness, precision, and applications. In this review, we first describe the function and applications of the CRISPR-Cas9 system. We next delineate how this technology could be utilized for gene therapy of various human disorders, including cancer and infectious diseases and highlight the promising examples in the field. Finally, we document current challenges and the potential solutions to overcome these obstacles for the effective use of CRISPR-Cas9 in clinical practice.

The demand for astaxanthin has been increasing for many health applications ranging from pharmaceuticals, food, cosmetics, and aquaculture due to its bioactive properties. Haematococcus pluvialis is widely recognized as the microalgae species with the highest natural accumulation of astaxanthin, which has made it a valuable source for industrial production. Astaxanthin produced by other sources such as chemical synthesis or fermentation are often produced in the cis configuration, which has been shown to have lower bioactivity. Additionally, some sources of astaxanthin, such as shrimp, may denature or degrade when exposed to high temperatures, which can result in a loss of bioactivity. Producing natural astaxanthin through the cultivation of H. pluvialis is presently a demanding and time-consuming task, which incurs high expenses and restricts the cost-effective industrial production of this valuable substance. The production of astaxanthin occurs through two distinct pathways, namely the cytosolic mevalonate pathway and the chloroplast methylerythritol phosphate (MEP) pathway. The latest advancements in enhancing product quality and extracting techniques at a reasonable cost are emphasized in this review. The comparative of specific extraction processes of H. pluvialis biological astaxanthin production that may be applied to large-scale industries were assessed. The article covers a contemporary approach to optimizing microalgae culture for increased astaxanthin content, as well as obtaining preliminary data on the sustainability of astaxanthin production and astaxanthin marketing information.

Osteoarthritis (OA) is a common degenerative joint disorder that adversely affects the quality of life of patients. Identification of novel diagnostic biomarkers is pivotal for the early detection and prevention of OA. Dataset GSE185059 was selected from Gene Expression Omnibus database to obtain differentially expressed lncRNAs (DE-lncRNAs), mRNAs (DE-mRNAs), and circRNAs (DE-circRNAs) between OA and normal samples. The Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses as well as protein-protein interaction (PPI) network construction of DE-mRNAs were conducted. Hub genes were identified from PPI networks and validated by RT-qPCR. starBase database was utilized for predicting miRNAs binding with hub genes, selected DE-lncRNAs and DE-circRNAs, respectively. The competing endogenous RNA (ceRNA) networks were constructed. A total of 818 DE-mRNAs, 191 DE-lncRNAs, and 2053 DE-circRNAs were identified. The DE-mRNAs were significantly enriched in several inflammation-related GO terms and KEGG pathways such as positive regulation of cell-cell adhesion, TNF-alpha signaling pathway and NF-kappa B signaling pathway. Thirteen hub genes were identified, which were CFTR, GART, SMAD2, NCK1, TJP1, UBE2D1, EFTUD2, PRKACB, IL10, SNRPG, CHD4, RPS24, and SRSF6. OA-related DE-lncRNA/circRNA-miRNA-hub gene networks were constructed. We identified 13 hub genes and constructed the ceRNA networks related to OA, providing a theoretical basis for further research.

Transfection efficiency of the immortalized human breast epithelial cell line MCF-10A remains an issue that needs to be resolved. In this study, it was aimed to deliver a recombinant DNA (pCMV-Azu-GFP) to the MCF-10A cells by the magnetofection method using magnetic nanoparticles (MNPs) and a simple magnet to accelerate the DNA delivery. Surface positively modified silica-coated iron oxide MNPs (MSNP-NH2) were produced and characterized via TEM, FTIR, and DLS analyses. The recombinant DNA (rDNA) was obtained by the integration of codon-optimized azurin to produce a fusion protein. Then, rDNA cloned in Escherichia coli cells was validated by sequence analysis. The electrostatically conjugated rDNA on MSNP-NH2 with an enhancer polyethyleneimine (PEI) was studied by agarose gel electrophoresis and the optimum conditions were determined to apply to the cell. A dose-dependent statistical difference was observed on treated cells based on the MTS test. The expression of the fusion protein after magnetofection was determined using laser scanning confocal microscope imaging and western blot analysis. It was observed that the azurin gene could be transferred to MCF-10A cells by magnetofection. Thus, when the azurin gene is used as a breast cancer treatment agent, it can be expressed in healthy cells without toxic effects.

Biliary atresia (BA) is a rare neonatal cholestatic disease that presents with a marked bile duct reaction and rapid fibrotic development. Our earlier research has shown that circUTRN24 is highly elevated in BA, but the exact molecular mechanism is still unknown. This study attempted to investigate whether circUTRN24 induces BA liver fibrosis through regulation of autophagy and to elucidate its molecular mechanism. Using TGF-β-treated hepatic stellate cells (HSC) LX-2, we created a liver fibrosis model. qRT-PCR was used to analyze the expression of circUTRN24, miR-483-3p, and IGF-1. Western blot analysis was used to assess the expression of IGF-1, HSC activation-related proteins, and autophagy-related proteins. The TGF-β-induced LX-2 cell fibrosis model was then supplemented with circUTRN24 siRNA, miR-483-3p mimics, and the autophagy activator Rapamycin, and functional rescue tests were carried out to investigate the role of circUTRN24, miR-483-3p, and autophagy in BA liver fibrosis. Using a luciferase reporter assay, a direct interaction between miR-483-3p and circUTRN24 or IGF-1 was discovered. With the increase of TGF-β treatment concentration, circUTRN24 expression also gradually increased, as did HSC activation and autophagy-related protein. si-circUTRN24 significantly decreased circUTRN24 expression and inhibited HSC activation and autophagy, which was reversed by Rapamycin. Through bioinformatics prediction and validation, we found circUTRN24 might act through miR-483-3p targeting IGF-1 in the autophagy-related mTOR pathway. Furthermore, miR-483-3p mimics significantly increased miR-483-3p expression and inhibited HSC activation and autophagy, which were reversed by Rapamycin. Functional rescue experiments showed that si-circUTRN24 inhibited circUTRN24 and IGF-1 expressions and promoted miR-483-3p expression, while the miR-483-3p inhibitor abolished these effects. These findings imply that circUTRN24/miR-483-3p/IGF-1 axis mediated LX-2 cell fibrosis by regulating autophagy.

Accumulating articles have reported the coding potential of long non-coding RNAs (lncRNAs). However, only a few lncRNAs-encoded peptides have been studied. Breast cancer (BRCA) progression-related gene modules were determined by weighted gene co-expression network analysis (WGCNA). Cell viability, proliferation, and migration capacities were assessed by Cell counting kit-8 (CCK8), 5-ethynyl-2'-deoxyuridine (EdU), and transwell assays. Immunofluorescence (IF) assay was implemented to observe protein expression. Co-immunoprecipitation (Co-IP) and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) were employed to analyze MAGI2 antisense RNA 3 (MAGI2-AS3)-ORF5-interacted proteins. WGCNA identified that MEpurple and MEblack modules were significantly negatively correlated with T stage in BRCA patients. MAGI2-AS3 was screened as one of the differentially expressed (DE) lncRNAs with translational potential in MEblack and MEpurple modules in BRCA. The data in The Cancer Genome Atlas (TCGA) uncovered that MAGI2-AS3 abundance was significantly decreased in invasive BRCA patients, and it had high diagnostic and prognostic values. MAGI2-AS3-ORF5 notably restrained BRCA cell viability, proliferation, and migration. Mechanically, MAGI2-AS3-ORF5 might affect the progression of BRCA cells by binding to extracellular matrix (ECM)-related proteins. MAGI2-AS3-ORF5 played an anti-tumor role by inhibiting BRCA cell viability, proliferation, and migration. MAGI2-AS3-ORF5 might modulate BRCA cell migration through ECM-associated proteins.

Staphylococcus aureus (S. aureus), as a Gram-positive bacterium, is commonly encountered in various infectious diseases, such as acute skin and soft tissue infections. Despite that many efforts have been made, sensitive and reliable quantitative determination of S. aureus remains a huge challenge. Here, we depict a novel colorimetric approach for sensitive and accurate detection by combining allosteric probe-based target recognition and chain extension-based dual signal recycling. The single-strand DNA (ssDNA) products generated by the chain extension process lead to the liberation of G-quadruplex sequences, which can fold into active DNAzyme under the assistance of hemin. The active DNAzyme can work as peroxidase mimics to catalyze the 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS2-)-H2O2 system, causing the color change of the system. Eventually, the method exhibits a wide detection range from 103 cfu/mL to 106 cfu/mL. The limit of detection of the approach was determined 232 cfu/mL. Considering the robust capability of the approach in S. aureus detection, we believe that it will be a potential alternative tool for biomedical research and clinical molecular diagnostics.

Recently, a new signaling complex Death-Associated Protein Kinase 1 (DAPK1)-N-methyl D-aspartate receptor subtype 2B (NR2B) engaged in the neuronal death cascade was identified where it was found that after stroke injury, N-methyl-D-aspartate glutamate (NMDA) receptors interact with DAPK1 through NR2B subunit and lead to excitotoxicity via overactivation of NMDA receptors. In this study, we used ZINC-12 database to find out potential inhibitor of DAPK1 and found some natural compounds showing good binding affinity towards DAPK1. These natural compounds showed interactions with ATP-binding site residues as well as substrate-recognition motifs. Thus, it has been concluded that the ligands those are showing interactions with both the sites could be considered as potential inhibitors for DAPK1.

Human umbilical cord mesenchymal stem cell (hucMSC)-derived exosomes (Exo) have been frequently investigated for disease control. This study was designed to explore the effects of hucMSC-Exo carrying lncRNA family with sequence similarity 99-member B (Exo-lncRNA FAM99B) on hepatocellular carcinoma (HCC) cell behaviour. The expression of lncRNA FAM99B in HCC cells was measured by reverse-transcription quantitative polymerase chain reaction. Protein levels of exosomal markers were quantified using western blotting. Flow cytometry analyses were performed to detect surface markers of hucMSCs and to measure the effects of Exo-lncRNA FAM99B on HCC cell cycle progression and cell apoptosis. Nanoparticle tracking analysis was used to measure the particle size of the exosomes. Additionally, cell viability was evaluated using methyl thiazolyl tetrazolium assays, and Transwell assays were performed to measure cell migration and invasion. Xenograft tumor models were established to explore the role of Exo-lncRNA FAM99B in vivo. Experimental results revealed that lncRNA FAM99B was downregulated in HCC cell lines, and low level of FAM99B is associated with poor survival rates in patients with HCC according to bioinformatics analysis. HucMSCs were identified in a good morphology with positively expressed CD105, CD29, and CD44 as well as negatively expressed CD31, CD14, and HLA-DR. High protein levels of exosomal markers (Alix, CD63 and TSG101) identified the existence of HucMSC-Exo. Importantly, the hucMSCs-Exo could enter HCC cells and exerted a suppressive effect on malignant cell activities. Moreover, overexpression of Exo-lncRNA FAM99B enhanced cell cycle arrest and cell apoptosis while suppressing cell viability, migration, and invasion in HCC. Exo-siRNA-FAM99B exerted the opposite effects on HCC cell process. In vivo experiments verified that Exo-lncRNA FAM99B inhibited tumorigenesis in HCC. In summary, lncRNA FAM99B derived from hucMSC-Exo inhibited malignant cellular phenotypes and tumorigenesis in HCC, which might provide a novel therapeutic strategy for HCC treatment.

Photoenzymatic decarboxylation shows great promise as a pathway for the generation of hydrocarbon fuels. CvFAP, which is derived from Chlorella variabilis NC64A, is a photodecarboxylase capable of converting fatty acids into hydrocarbons. CvFAP is an example of coupling biocatalysis and photocatalysis to produce alkanes. The catalytic process is mild, and it does not yield toxic substances or excess by-products. However, the activity of CvFAP can be readily inhibited by several factors, and further enhancement is required to improve the enzyme yield and stability. In this article, we will examine the latest advancements in CvFAP research, with a particular focus on the enzyme's structural and catalytic mechanism, summarized some limitations in the application of CvFAP, and laboratory-level methods for enhancing enzyme activity and stability. This review can serve as a reference for future large-scale industrial production of hydrocarbon fuels.

Nanotechnology and nanostructured materials for drug delivery and tissue engineering applications are relatively new field that is constantly advancing and expanding. The materials used are at the nanoscale level. Recently, great discoveries and applications have been made (Agents for use in chemotherapy, biological agents and immunotherapy agents) in the treatment of diseases in various areas. Tissue engineering is based on the regeneration and repair of damaged organs and tissues by developing biological substitutes that restore, maintain or improve the function of tissues and organs. Cells isolated from patients are used to seed 3D nanoparticles that can be synthetic or natural biomaterials. For the development of new tissue in tissue engineering, it is necessary to meet the conditions for connecting cells. This paper will present the ways of connecting cells and creating new tissues. Some recent discoveries and advances in the field of nanomedicine and the application of nanotechnology in drug delivery will be presented. Furthermore, the improvement of the effectiveness of new and old drugs based on the application of nanotechnology will be shown.

Cellular RNAs, both coding and noncoding are adorned by > 100 chemical modifications, which impact various facets of RNA metabolism and gene expression. Very often derailments in these modifications are associated with a plethora of human diseases. One of the most oldest of such modification is pseudouridylation of RNA, wherein uridine is converted to a pseudouridine (Ψ) via an isomerization reaction. When discovered, Ψ was referred to as the 'fifth nucleotide' and is chemically distinct from uridine and any other known nucleotides. Experimental evidence accumulated over the past six decades, coupled together with the recent technological advances in pseudouridine detection, suggest the presence of pseudouridine on messenger RNA, as well as on diverse classes of non-coding RNA in human cells. RNA pseudouridylation has widespread effects on cellular RNA metabolism and gene expression, primarily via stabilizing RNA conformations and destabilizing interactions with RNA-binding proteins. However, much remains to be understood about the RNA targets and their recognition by the pseudouridylation machinery, the regulation of RNA pseudouridylation, and its crosstalk with other RNA modifications and gene regulatory processes. In this review, we summarize the mechanism and molecular machinery involved in depositing pseudouridine on target RNAs, molecular functions of RNA pseudouridylation, tools to detect pseudouridines, the role of RNA pseudouridylation in human diseases like cancer, and finally, the potential of pseudouridine to serve as a biomarker and as an attractive therapeutic target.

The constant increase in demand for food, valued bio-based compounds and energy demand has prompted the development of innovative and sustainable resources. New technologies and strategies must be implemented to boost microalgae biomass production, such as using different photoperiods along with (LED) light-emitting diodes to stimulate biomass production and boost profits. This work investigates the cultivation of blue-green microalgae (Spirulina) in a closed lab condition. The current study aims to boost Spirulina biomass production by creating ideal growth conditions using different photoperiods (12:12; 10:14; 14:10) light/dark with a constant light intensity of 2000 lx from White LED lights. The obtained optical density and protein content was highest for photoperiod 14L: 10D and values were 0.280 OD, with a protein content of 23.44 g/100 g, respectively. This study is a crucial first step in identifying the best photoperiod conditions to help S. platensis produce more biomass. The study results showed that increasing photoperiod for S. platensis farming can improve the quality and amount of biomass generated in those cultures without negatively affecting growth.

Chagas disease-caused by the parasite Trypanosoma cruzi-is a neglected tropical disease for which available drugs are not fully effective in the chronic stage and a vaccine is not available yet. Microalgae represent a promising platform for the production and oral delivery of low-cost vaccines. Herein, we report a vaccine prototype against T. cruzi produced in a microalgae platform, based on the candidate antigen Tc24 with a C terminus fusion with the Co1 peptide (Tc24:Co1 vaccine prototype). After modeling the tertiary structure, in silico studies suggested that the chimeric protein is antigenic, not allergenic, and molecular docking indicated binding with Toll-like receptors 2 and 4. Thus, Tc24:Co1 was expressed in the marine microalga Schizochytrium sp., and Western blot confirmed the expression at 48 h after induction, with a yield of 632 µg/L of algal culture (300 μg/g of lyophilized algal cells) as measured by the enzyme-linked immunosorbent assay (ELISA). Upon oral administration of whole-cell Schizochytrium sp. expressing Tc24:Co1 (7.5 µg or 15 µg of Tc24:Co1 doses) in mice, specific serum IgG and intestinal mucosa IgA responses were detected in addition to an increase in serum Th1/Th2 cytokines. In conclusion, Schizochytrium sp.-expressing Tc24:Co1 is a promising oral vaccine prototype to be evaluated in an animal model of Trypanosoma cruzi infection.

Agrobacterium tumefaciens-mediated plant transformation is the most dominant technique for the transformation of plants. It is used to transform monocotyledonous and dicotyledonous plants. A. tumefaciens apply for stable and transient transformation, random and targeted integration of foreign genes, as well as genome editing of plants. The Advantages of this method include cheapness, uncomplicated operation, high reproducibility, a low copy number of integrated transgenes, and the possibility of transferring larger DNA fragments. Engineered endonucleases such as CRISPR/Cas9 systems, TALENs, and ZFNs can be delivered with this method. Nowadays, Agrobacterium-mediated transformation is used for the Knock in, Knock down, and Knock out of genes. The transformation effectiveness of this method is not always desirable. Researchers applied various strategies to improve the effectiveness of this method. Here, a general overview of the characteristics and mechanism of gene transfer with Agrobacterium is presented. Advantages, updated data on the factors involved in optimizing this method, and other useful materials that lead to maximum exploitation as well as overcoming obstacles of this method are discussed. Moreover, the application of this method in the generation of genetically edited plants is stated. This review can help researchers to establish a rapid and highly effective Agrobacterium-mediated transformation protocol for any plant species.

Beta-thalassemia is one of the most common monogenic inherited disorders worldwide caused by different mutations in the hemoglobin subunit beta (HBB) gene. Genome-editing based on clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 system (CRISPR/Cas9) has raised the hope for life-long gene therapy of beta-thalassemia. In a proof-of-concept study, we describe the detailed design and assess the efficacy of a novel homology-directed repair (HDR)-based CRISPR construct for targeting the HBB locus. The selected sgRNAs were designed and cloned into an optimized CRISPR plasmid. The HDR donor templates containing a reporter and a selection marker flanked by the piggyBac Inverted Tandem Repeat (ITRs), the homology arms and the delta thymidine kinase (ΔTK) gene for negative selection were constructed. The efficiency of on-target mutagenesis by the eSpCas9/sgRNAs was assessed by mismatch assays. HDR-positive cells were isolated by treatment with G418 or selection based on truncated Neuron Growth Factor Receptor (tNGFR) expression using the Magnetic Activated Cell Sorting (MACS) method followed by ganciclovir (GCV) treatment to eliminate cells with random genomic integration of the HDR donor template. In-out PCR and sanger sequencing confirmed HDR in the isolated cells. Our data showed ~ 50% efficiency for co-transfection of CRISPR/donor template plasmids in HEK293 cells and following G418 treatment, the HDR efficiency was detected at ~ 37.5%. Moreover, using a clinically-relevant strategy, HDR events were validated after selection for tNGFR+ cells followed by negative selection for ΔTK by GCV treatment. Thus, our HDR-based gene-editing strategy could efficiently target the HBB locus and enrich for HDR-positive cells.

MicroRNAs (miRNAs) are small single-stranded regulatory RNAs that are shown to be dysregulated in a wide array of human cancers. MiRNAs play critical roles in cancer progression and function as either oncogenes or tumor suppressors through modulating various target genes. Therefore, they possess great potential as diagnostic and therapeutic targets for cancer detection and treatment. In particular, recent studies have illustrated that miR-425 is also dysregulated in various human malignancies and plays a fundamental role in cancer initiation and progression. miR-425 has been reported to function as a dual-role miRNA participating in the regulation of cellular processes, including metastasis, invasion, and cell proliferation by modulating multiple signaling pathways, such as TGF-β, Wnt, and P13K/AKT pathways. Therefore, regarding recent researches showing the high therapeutic potential of miR-425, in this review, we have noted the impact of its dysregulation on signaling pathways and various aspects of tumorigenesis in a variety of human cancers.

The central nervous system (CNS) is a complicated neural network. The origin and evolution of functional neurons and glia cells remain unclear, as do the cellular alterations that occur during the course of cerebral disease rehabilitation. Lineage tracing is a valuable method for tracing specific cells and achieving a better understanding of the CNS. Recently, various technological breakthroughs have been made in lineage tracing, such as the application of various combinations of fluorescent reporters and advances in barcode technology. The development of lineage tracing has given us a deeper understanding of the normal physiology of the CNS, especially the pathological processes. In this review, we summarize these advances of lineage tracing and their applications in CNS. We focus on the use of lineage tracing techniques to elucidate the process CNS development and especially the mechanism of injury repair. Deep understanding of the central nervous system will help us to use existing technologies to diagnose and treat diseases.

There is a need for an efficient and long-lasting treatment due to the population's increasing prevalence of neurodegenerative disorders. In an effort to generate fresh ideas and create novel therapeutic medications, scientists have recently started to investigate the biological functions of compounds derived from plants and herbs. Ginseng, famous Chinese herbal medicine, has therapeutic value by virtue of its compounds ginsenosides or panaxosides, which are triterpene saponins and steroid glycosides. Research revealed positive impacts on ameliorating various disease conditions and found it as a possible drug candidate. Several neuroprotection mechanisms followed by this compound are inhibition of cell apoptosis, oxidative stress, inflammatory, and tumor activity. It has been demonstrated that controlling these mechanisms enhances cognitive performance and safeguards the brain against neurodegenerative disorders. The main objective of this review is to give a description of the most recent studies on ginsenoside's possible therapeutic application in the treatment of neurodegenerative diseases. Using organic compounds like ginseng and its various components may create new avenues for innovative treatment approaches development for neurological diseases. However, further research is necessary to confirm the stability and effectiveness of ginsenosides for neurodegenerative disease.

Single-cell RNA-seq (scRNA-seq) is a revolutionary technology that allows for the genomic investigation of individual cells in a population, allowing for the discovery of unusual cells associated with cancer and metastasis. ScRNA-seq has been used to discover different types of cancers with poor prognosis and medication resistance such as lung cancer, breast cancer, ovarian cancer, and gastric cancer. Besides, scRNA-seq is a promising method that helps us comprehend the biological features and dynamics of cell development, as well as other disorders. This review gives a concise summary of current scRNA-seq technology. We also explain the main technological steps involved in implementing the technology. We highlight the present applications of scRNA-seq in cancer research, including tumor heterogeneity analysis in lung cancer, breast cancer, and ovarian cancer. In addition, this review elucidates potential applications of scRNA-seq in lineage tracing, personalized medicine, illness prediction, and disease diagnosis, which reveals that scRNA-seq facilitates these events by producing genetic variations on the single-cell level.

LncRNA ZNF667-AS1 plays an important role in the carcinogenesis and progression of various cancers. However, their role in colon cancer (CC) remains unclear. The expression of ZNF667-AS1, KIF5C, and miR-523-3p in CC cells and tissues was analyzed using RT-qPCR and western blotting. CCK-8 scratch-wound assay, western blotting, and flow cytometry were conducted to investigate the malignant activity of CC in vitro. Luciferase reporter, RNA pull-down, and Ago2 immunoprecipitation (RIP) experiments were conducted to ascertain the association of miR-523-3p with ZNF667-AS1 and KIF5C 3'UTR. Xenograft tumor experiments were also performed. CC cells and tissues showed low expression of NF667-AS1 and KIF5C and elevated expression of miR-523-3p. ZNF667-AS1 overexpression attenuates proliferation and migration of CC cells, restores inactivated apoptosis in vitro, and inhibits tumor growth in vivo. MiR-523-3p targets both ZNF667-AS1 and the KIF5C 3'UTR. ZNF667-AS1 overexpression in SW480 and SW620 cells attenuated the oncogenic effect of miR-523-3p in CC. However, this attenuating effect was counteracted by KIF5C overexpression. ZNF667-AS1 sequestered miR-523-3, reducing miR-523-3p-mediated inhibition of KIF5C expression, thereby repressing colon carcinogenesis in vitro. Our findings shed light on a novel anticancer strategy that could potentially combat CC.

Ovarian cancer (OC) is among several general malignant gynecological cancers associated with high mortality rates on a global scale. Earlier investigations have revealed a critical role of circular RNAs (circRNAs) in OC development, which is a new class of endogenous non-coding RNA (ncRNA) that reported to mediate progression of diverse tumor types. At present, the precise involvement of circRNAs and associated regulatory mechanisms in OC remain unknown. In this study, hsa_circ_0001741 expression patterns in OC cells and tissues were tested. The underlying regulatory pathways and targets were further explored with the aid of bioinformatics, luciferase reporter, 5-ethynyl-2'-deoxyuridine (EdU) and cell counting kit-8 (CCK-8) analyses. Further investigation of the hsa_circ_0001741 effects on tumor growth in vivo revealed abnormal circRNA expression in OC. hsa_circ_0001741 expression reduced in OC cells and tissues, indicative of activity in OC progression. hsa_circ_0001741 upregulation resulted in OC proliferation inhibitions. The luciferase reporter outputs verified miR-188-5p and FOXN2 as hsa_circ_0001741 downstream targets. FOXN2 silencing or miR-188-5p upregulations reversed inhibitory effects regarding hsa_circ_0001741 on OC cell proliferation. Therefore our data suggested that hsa_circ_0001741 upregulation inhibited proliferation of OC through modulatory effects on miR-188-5p/FOXN2 signaling.

This study investigated the mechanism of neurotrophin-3 (NT-3) in promoting spinal cord injury repair through the transforming growth factor-beta (TGF-β) signaling pathway. A mouse model of spinal cord injury was established. Forty C57BL/6J mice were randomized into model, NT-3, NT-3 + TGF-β1 and NT-3 + LY364947 groups. The Basso-Beattie-Bresnahan (BBB) scores of the NT-3 and NT-3 + LY364947 groups were significantly higher than the model group. The BBB score of the NT-3 + TGF-β1 group was significantly lower than NT-3 group. Hematoxylin-eosin staining and transmission electron microscopy showed reduction in myelin sheath injury, more myelinated nerve fibers in the middle section of the catheter, and relatively higher density and more neatly arranged regenerated axons in the NT-3 and NT-3 + LY364947 groups compared with the model and NT-3 + TGF-β1 groups. Immunofluorescence, TUNEL and Western blot analysis showed that compared with model group, the NEUN expression increased, and the apoptosis and Col IV, LN, CSPG, tenascin-C, Sema 3 A, EphB2 and Smad2/3 protein expression decreased significantly in the NT-3 and NT-3 + LY364947 groups; the condition was reversed in the NT-3 + TGF-β1 group compared with the NT-3 group. NT-3 combined with TGF-β signaling pathway promotes astrocyte differentiation, reduces axon regeneration inhibitory molecules, apoptosis and glial scar formation, promotes axon regeneration, and improves spinal cord injury.

Psidium guajava fruits are highly appreciated for their nutrients and bioactive compounds content, which contribute to their antioxidant and antimicrobial capacities. The purpose of this study was to determine bioactive compound (phenolic, flavonoids, and carotenoid contents), antioxidant activity (DPPH, ABTS, ORAC, and FRAP), and antibacterial potential against MDR and food-borne pathogenic strains of Escherichia coli, and Staphylococcus aureus during different stages of fruit ripening.The results elucidated that ripe fruits (methanolic extract) contain the highest total phenolic, flavonoids, and carotenoid contents (417.36 ± 2.63 µg GAE/gm of FW, 711.78 ± 0.70 µg QE/gm of FW and 0.683 ± 0.06 µg/gm of FW) followed by hexane, ethyl acetate, and aqueous. Methanolic extract of the ripe fruits showed the highest antioxidant activity when measured by DPPH (61.55 ± 0.91%), FRAP (31.83 ± 0.98 mM Fe(II)/gm of FW), ORAC (17.19 ± 0.47 mM TE/ gm of FW), and ABTS (41.31 ± 0.99 µmol Trolox/gm of FW) assays. In the antibacterial assay, the ripe stage had the highest antibacterial activity against MDR and food-borne pathogenic strains of Escherichia coli, and Staphylococcus aureus. The methanolic ripe extract was found to possess maximum antibacterial activity ZOI, MIC, and IC50 18.00 ± 1.00 mm, 95.95 ± 0.05%, and 0.58 μg/ml; 15.66 ± 0.57 mm, 94.66 ± 0.19%, and 0.50 μg/ml, respectively, against pathogenic and MDR strains of E. coli and 22.33 ± 0.57 mm, 98.97 ± 0.02%, and 0.26 μg/ml; 20.33 ± 1.15 mm, 96.82 ± 0.14%, and 0.39 μg/ml, respectively, against pathogenic and MDR strains of S. aureus. Considering the bioactive compounds and beneficial effects, these fruit extracts could be promising antibiotic alternatives, avoiding antibiotic overuse and its negative effects on human health and the environment, and can be recommended as a novel functional food.

The ability to detect early metabolic changes in patients who have an increased mortality risk in the intensive care units (ICUs) could increase the likelihood of predicting recovery patterns and assist in disease management. Markers that can predict the disease progression of patients in the ICU might also be beneficial for improving their medical profile. Although biomarkers have been used in the ICU more frequently in recent years, the clinical use of most of them is limited. A wide range of biological processes are influenced by microRNAs (miRNAs) that modulate the translation and stability of specific mRNAs. Studies suggest that miRNAs may serve as a diagnostic and therapeutic biomarker in ICUs by profiling miRNA dysregulation in patient samples. To improve the predictive value of biomarkers for ICU patients, researchers have proposed both investigating miRNAs as novel biomarkers and combining them with other clinical biomarkers. Herein, we discuss recent approaches to the diagnosis and prognosis of patients admitted to an ICU, highlighting the use of miRNAs as novel and robust biomarkers for this purpose. In addition, we discuss emerging approaches to biomarker development and ways to improve the quality of biomarkers so that patients in ICU get the best outcomes possible.

Staphylea holocarpa (Hemsley 1895) is an ornamental deciduous shrub or tree in the family Staphyleaceae. As the shortage of the wild resources, S. holocarpa is also a rare plant. The revelation of the species origin and evolution progress and the relation. Therefore, the S. holocarpa complete chloroplast genome sequence was completed and characterized by de novo assembly. The cp genome length of S. holocarpa was 160,461 bp and it has a typical quadripartite structure, consisted of an 89,760 bp large single-copy region and a 18,639 bp small single-copy region, which were divided by two inverted repeat regions of 26,031 bp. After genome annotation, it comes to 130 genes that were predicted, which includes 85, 8, and 37 encoded proteins, rRNA, and tRNA, respectively. A phylogenetic analysis has shown that the S. holocarpa cp genome is related to the Staphylea trifolia. This work will be useful for further population genomic and phylogenetic studies of S. holocarpa.