Toxicodynamic of combined mycotoxins: MicroRNAs and acute-phase proteins as diagnostic biomarkers.

Mycotoxins, ubiquitous contaminants in food, present a global threat to human health and well-being. Mitigation efforts, such as the implementation of sound agricultural practices, thorough food processing, and the advancement of mycotoxin control technologies, have been instrumental in reducing mycotoxin exposure and associated toxicity. To comprehensively assess mycotoxins and their toxicodynamic implications, the deployment of effective and predictive strategies is imperative. Understanding the manner of action, transformation, and cumulative toxic effects of mycotoxins, moreover, their interactions with food matrices can be gleaned through gene expression and transcriptome analyses at cellular and molecular levels. MicroRNAs (miRNAs) govern the expression of target genes and enzymes that play pivotal roles in physiological, pathological, and toxicological responses, whereas acute phase proteins (APPs) exert regulatory control over the metabolism of therapeutic agents, both endogenously and posttranscriptionally. Consequently, this review aims to consolidate current knowledge concerning the regulatory role of miRNAs in the initiation of toxicological pathways by mycotoxins and explores the potential of APPs as biomarkers following mycotoxin exposure. The findings of this research highlight the potential utility of miRNAs and APPs as indicators for the detection and management of mycotoxins in food through biological processes. These markers offer promising avenues for enhancing the safety and quality of food products.

Cold plasma: A success road to mycotoxins mitigation and food value edition.

Mycotoxins are common in many agricultural products and may harm both animals and humans. Dietary mycotoxins are reduced via physical, chemical, and thermal decontamination methods. Chemical residues are left behind after physical and chemical treatments that decrease food quality. Since mycotoxins are heat-resistant, heat treatments do not completely eradicate them. Cold plasma therapy increases food safety and shelf life. Cold plasma-generated chemical species may kill bacteria quickly at room temperature while leaving no chemical residues. This research explains how cold plasma combats mold and mycotoxins to guarantee food safety and quality. Fungal cells are damaged and killed by cold plasma species. Mycotoxins are also chemically broken down by the species, making the breakdown products safer. According to a preliminary cold plasma study, plasma may enhance food shelf life and quality. The antifungal and antimycotoxin properties of cold plasma benefit fresh produce, agricultural commodities, nuts, peppers, herbs, dried meat, and fish.

Identification and function of the Pax gene Bmgsb in the silk gland of Bombyx mori.

Paired box (Pax) genes are highly conserved throughout evolution, and the Pax protein is an important transcription factor of embryonic development. The Pax gene Bmgsb is expressed in the silk glands of silkworm, but its biological functions remain unclear. This study aimed to investigate the expression pattern of Bmgsb in the silk gland and explore its functions using RNA interference (RNAi). Here, we identified eight Pax genes in Bombyx mori. Phylogenetic analysis showed that the B. mori Pax genes were highly homologous to the Pax genes in other insects and highly evolutionarily conserved. The tissue expression profile showed that Bmgsb was expressed in the anterior silk gland and anterior part of the middle silk gland (AMSG). RNAi of Bmgsb resulted in defective development of the AMSG, and the larvae were mostly unable to cocoon in the wandering stage. RNA-seq analysis showed that the fibroin genes fib-l, fib-h and p25, cellular heat shock response-related genes and phenol oxidase genes were considerably upregulated upon Bmgsb knockdown. Furthermore, quantitative reverse transcription-PCR results showed that the fibroin genes and ubiquitin proteolytic enzyme-related genes were significantly upregulated in the AMSG after Bmgsb knockdown. This study provides a foundation for future research on the biological functions of B. mori Pax genes. In addition, it demonstrates the important roles of Bmgsb in the transcriptional regulation of fibroin genes and silk gland development.

Molecular cloning, biological description, and functional analysis of Ajfos transcription factor in pathogen-induced Apostichopus japonicus.

Activator protein-1 subfamily member c-Fos wields significant influence over cellular activities, such as regulation of cell growth and division, cell death, and immune responses under various extracellular situations. In this study, the full-length c-Fos of sea cucumber, Apostichopus japonicus (Ajfos) was successfully cloned and analyzed. The anticipated 306 amino acid sequences of Ajfos displayed a basic-leucine zipper (bZIP) domain, similar to invertebrate counterparts. In addition, the qPCR results suggested Ajfos expressed in all tissues, with the highest level in coelomocytes from polian vesicle (vesicle lumen cells), followed by coelomocytes from coelom (coelomocytes). Moreover, the expression levels of Ajfos in the coelomocytes and vesicle lumen cells of sea cucumber showed significant changes after the Vibrio splendidus challenge, especially reaching a peak at 6 h. Compared with the silencing negative control RNA interference (siNC) group, silencing Ajfos (siAjfos) in vivo decreased the downstream proliferation-related gene expression of vesicle lumen cells after infection with V. splendidus while no significant influence was observed on coelomocytes. Furthermore, the proliferation proportion of vesicle lumen cells in the siAjfos group was significantly reduced under pathogen stimulation conditions. Finally, based on the fluctuation trend of total coelomocyte density (TCD) from coelom and polian vesicle previously discovered, it is evident that Ajfos played a critical role in facilitating the swift proliferation of vesicle lumen cells in response to V. splendidus stimulation. Altogether, this research provided an initial reference of the function of Ajfos in echinoderms, unveiling its participation in host coelomocyte proliferation of sea cucumbers during bacterial challenges.

Evaluating the efficacy of auricular acupuncture for chemotherapy-induced taste alterations: A pilot randomized controlled trial.

PURPOSE: Irrespective of the development of acupuncture-based interventions, clinical evidence regarding their efficacy remains controversial owing to issues with the study design and an unclear risk of bias. This study aimed to evaluate the efficacy of auricular acupuncture in managing taste alterations in patients with cancer undergoing platinum-based chemotherapy. METHODS: We conducted a pilot randomized controlled trial involving 73 patients randomly assigned to an auricular acupuncture or a control group. The primary outcome was the severity of chemotherapy-induced taste alterations, and the secondary outcomes included quality of life and negative emotions of the patients. RESULTS: A total of 49 participants completed the study. Compared to the control group, patients in the auricular acupuncture group showed significant reductions in discomfort, general taste alterations, and total scores on the Chemotherapy-induced Taste Alteration Scale (all p < 0.05). Furthermore, we observed significant improvements in quality of life, including physical function (p = 0.007), role function (p = 0.006), emotional function (p = 0.016), nausea and vomiting (p = 0.021), appetite loss (p = 0.046), and significant improvements in anxiety and depression (p < 0.01). CONCLUSIONS: Our findings suggest that auricular acupuncture may be a beneficial intervention for managing chemotherapy-induced taste alterations in patients with cancer receiving platinum-based chemotherapy. It may also contribute to improvements in quality of life and negative emotions. However, these results are preliminary, and further evaluation with larger randomized controlled trials is necessary.

KaScape: a sequencing-based method for global characterization of protein‒DNA binding affinity.

It is difficult to exhaustively screen all possible DNA binding sequences for a given transcription factor (TF). Here, we developed the KaScape method, in which TFs bind to all possible DNA sequences in the same DNA pool where DNA sequences are prepared by randomized oligo synthesis and the random length can be adjusted to a length such as 4, 5, 6, or 7. After separating bound from unbound double-stranded DNAs (dsDNAs), their sequences are determined by next-generation sequencing. To demonstrate the relative binding affinities of all possible DNA sequences determined by KaScape, we developed three-dimensional KaScape viewing software based on a K-mer graph. We applied KaScape to 12 plant TF family AtWRKY proteins and found that all AtWRKY proteins bound to the core sequence GAC with similar profiles. KaScape can detect not only binding sequences consistent with the consensus W-box "TTGAC(C/T)" but also other sequences with weak affinity. KaScape provides a high-throughput, easy-to-operate, sensitive, and exhaustive method for quantitatively characterizing the relative binding strength of a TF with all possible binding sequences, allowing us to comprehensively characterize the specificity and affinity landscape of transcription factors, particularly for moderate- and low-affinity binding sites.

Feasibility insights into the application of Paenibacillus pabuli E1 in animal feed to eliminate non-starch polysaccharides.

The goal of the research was to find alternative protein sources for animal farming that are efficient and cost-effective. The researchers focused on distillers dried grains with solubles (DDGS), a co-product of bioethanol production that is rich in protein but limited in its use as a feed ingredient due to its high non-starch polysaccharides (NSPs) content, particularly for monogastric animals. The analysis of the Paenibacillus pabuli E1 genome revealed the presence of 372 genes related to Carbohydrate-Active enzymes (CAZymes), with 98 of them associated with NSPs degrading enzymes that target cellulose, hemicellulose, and pectin. Additionally, although lignin is not an NSP, two lignin-degrading enzymes were also examined because the presence of lignin alongside NSPs can hinder the catalytic effect of enzymes on NSPs. To confirm the catalytic ability of the degrading enzymes, an in vitro enzyme activity assay was conducted. The results demonstrated that the endoglucanase activity reached 5.37 U/mL, while beta-glucosidase activity was 4.60 U/mL. The filter paper experiments did not detect any reducing sugars. The xylanase and beta-xylosidase activities were measured at 11.05 and 4.16 U/mL, respectively. Furthermore, the pectate lyase and pectin lyase activities were found to be 8.19 and 2.43 U/mL, respectively. The activities of laccase and MnP were determined as 1.87 and 4.30 U/mL, respectively. The researchers also investigated the effect of P. pabuli E1 on the degradation of NSPs through the solid-state fermentation of DDGS. After 240 h of fermentation, the results showed degradation rates of 11.86% for hemicellulose, 11.53% for cellulose, and 8.78% for lignin. Moreover, the crude protein (CP) content of DDGS increased from 26.59% to 30.59%. In conclusion, this study demonstrated that P. pabuli E1 possesses various potential NSPs degrading enzymes that can effectively eliminate NSPs in feed. This process improves the quality and availability of the feed, which is important for animal farming as it seeks alternative protein sources to replace traditional nutrients.

Transcriptome analysis reveals key transcription factors and pathways of polian vesicle associated with cell proliferation in Vibrio splendidus-challenged Apostichopus japonicus.

Polian vesicle is thought to produce coelomocytes and contribute to the sea cucumber's immune system. Our previous work has indicated that polian vesicle was responsible for cell proliferation at 72 h post pathogenic challenge. However, the transcription factors related to the activation of effector factors and the molecular process behind this remained unknown. In this study, to reveal the early functions of polian vesicle in response to the microbe, a comparative transcriptome sequencing of polian vesicle in V. splendidus-challenged Apostichopus japonicus, including normal group (PV 0 h), pathogen challenging for 6 h (PV 6 h) and 12 h (PV 12 h) was performed. Compared PV 0 h to PV 6 h, PV 0 h to PV 12 h, and PV 6 h to PV 12 h, we found 69, 211, and 175 differentially expressed genes (DEGs), respectively. KEGG enrichment analysis revealed the DEGs, including several transcription factors such as fos, FOS-FOX, ATF2, egr1, KLF2, and Notch3 between PV 6 h and PV 12 h were consistently enriched in MAPK, Apelin and Notch3 signaling pathways related to cell proliferation compared with that in PV 0 h. Important DEGs involved in cell growth were chosen, and their expression patterns were almost the same as the transcriptome profile analysis by qPCR. Protein interaction network analysis indicated that two DEGs of fos and egr1 were probably significant as key candidate genes controlling cell proliferation and differentiation in polian vesicle after pathogenic infection in A. japonicus. Overall, our analysis demonstrates that polian vesicles may play an essential role in regulating proliferation via transcription factors-mediated signaling pathway in A. japonicus and provide new insights into hematopoietic modulation of polian vesicles in response to pathogen infection.

Characterization of a Filamentous Phage, Vaf1, from Vibrio alginolyticus AP-1.

Filamentous phages are ubiquitously distributed in the global oceans. However, little is known about their biological contribution to their host's genetic and phenotypic diversity. In this study, a filamentous phage, Vaf1, was isolated and characterized from the emerging marine pathogen strain Vibrio alginolyticus AP-1. We explored the effects of the resident phage Vaf1 on the host physiology under diverse conditions by precisely deleting the entire phage Vaf1. Our results demonstrate that the presence of phage Vaf1 significantly increased biofilm formation, swarming motility, and contact-dependent competition. Furthermore, the gene expression profile suggests that several phage genes were upregulated in response to low-nutrient conditions. Unexpectedly, an in vivo study of zebrafish shows that fish infected with strain ΔVaf1 survived longer than those infected with wild-type strain AP-1, indicating that Vaf1 contributes to the virulence of V. alginolyticus. Together, our results provide direct evidence for the effect of Vaf1 phage-mediated phenotypic changes in marine bacteria V. alginolyticus. This further emphasizes the impressive complexity and diversity that filamentous phage-host interactions pose and the challenges associated with bacterial disease control in marine aquaculture. IMPORTANCE Non-lytic filamentous phages can replicate without killing their host, establishing long-term persistence within the bacterial host. In contrast to the well-studied CTXφ phage of the human-pathogenic Vibrio cholerae, little is known about the filamentous phage Vaf1 and its biological role in host fitness. In this study, we constructed a filamentous phage-deleted strain, ΔVaf1, and provided direct evidence on how an intact phage, φVaf1, belonging to the family Inoviridae, helps the bacterial host AP-1 to overcome adverse environmental conditions. Our results likely open new avenues for fundamental studies on how filamentous phage-host interactions regulate different aspects of Vibrio cell behaviors.

Protective efficacy of a recombinant enterotoxin antigen in a maternal immunization model and the inhibition of specific maternal antibodies to neonatal oral vaccination.

ETEC (enterotoxigenic Escherichia coli) infection is the leading cause of profuse watery diarrhea in mammals, especially among pre-weaning and post-weaning piglets in swine industry. Maternal immunization is a current rational strategy for providing protection to susceptive piglets and reducing the incidence of ETEC-associated diarrhea. Here we evaluated the protective efficiency of a recombinant antigen (MBP-SLS) fused by major enterotoxin subunits (STa-LTB-STb) via a maternal immunization model, and the impacts of maternal antibodies to neonatal oral vaccination were also investigated in the neonates. The high titers of specific IgG and sIgA in pups shown that the maternal antibodies could be transferred passively. Furthermore, the increases of IL-6 and IL-10 cytokines in breast milk and pup serum indicated that immunization on mother could effectively boost the immune system of neonates. Newborn rats from immunized mothers showed a 70% survival rate after ETEC infection. However, the mucosal immune responses of neonates were inhibited by the high level of maternal specific antibodies. Among the oral-vaccinated neonates, born from mock-immunized rats reached the highest survival after ETEC challenge. Collectively, the fusion MBP-SLS antigen could effectively induce strong immune responses in rats during pregnancy and the pups could receive passive protection through specific antibodies transferred via milk and placenta. However, the specific maternal antibodies exhibited an inhibition effect on the mucosal immune responses in offspring.

In-vitro assessment of a novel plant rhizobacterium, Citrobacter freundii, for degrading and biocontrol of food mycotoxin deoxynivalenol.

Deoxynivalenol (DON) is one of the most harmful and well-known toxins present in food and animal feed throughout the world. Citrobacter freundii (C. freundii-ON077584), a novel DON-degrading strain, was isolated from rice root-linked soil samples. The degrading properties, including DON concentrations, incubation pH, incubation temperatures, bacterial concentrations, and acid treatment effect on degradation, were evaluated. At pH 7 and an incubation temperature of 37 °C, C. freundii demonstrated the capability to degrade more than 90% of DON. The degraded products of DON were identified as 3-keto-DON and DOM-1, which were confirmed by High Performance Liquid Chromatography (HPLC) and Ultra-Performance Liquid Chromatography hyphenated with Tandem Mass Spectrometry (UPLC-MS/MS) analyses. The mechanism of DON degradation into 3-keto-DON and DOM-1 by this bacterial strain will be further explored to identify and purify novel degrading enzymes that can be cloned to the microorganism and added to the animal feed to degrade the DON in the digestion tract.

DNA-TCP complex structures reveal a unique recognition mechanism for TCP transcription factor families.

Plant-specific TCP transcription factors are key regulators of diverse plant functions. TCP transcription factors have long been annotated as basic helix-loop-helix (bHLH) transcription factors according to remote sequence homology without experimental validation, and their consensus DNA-binding sequences and protein-DNA recognition mechanisms have remained elusive. Here, we report the crystal structures of the class I TCP domain from AtTCP15 and the class II TCP domain from AtTCP10 in complex with different double-stranded DNA (dsDNA). The complex structures reveal that the TCP domain is a distinct DNA-binding motif and the homodimeric TCP domains adopt a unique three-site recognition mode, binding to dsDNA mainly through a central pair of ß-strands formed by the dimer interface and two basic flexible loops from each monomer. The consensus DNA-binding sequence for class I TCPs is a perfectly palindromic 11 bp (GTGGGNCCCAC), whereas that for class II TCPs is a near-palindromic 11 bp (GTGGTCCCCAC). The unique DNA binding mode allows the TCP domains to display broad specificity for a range of DNA sequences even shorter than 11 bp, adding further complexity to the regulatory network of plant TCP transcription factors.

A strategy of co-fermentation of distillers dried grains with solubles (DDGS) and lignocellulosic feedstocks as swine feed.

To meet the sustainable development of the swine feed industry, it is essential to find alternative feed resources and develop new feed processing technologies. Distillers dried grains with solubles (DDGS) is a by-product from the ethanol industry consisting of adequate nutrients for swine and is an excellent choice for the swine farming industry. Here, a strategy of co-fermentation of DDGS and lignocellulosic feedstocks for production of swine feed was discussed. The potential of the DDGS and lignocellulosic feedstocks as feedstock for fermented pig feed and the complementary relationship between them were described. In order to facilitate the swine feed research in co-fermentation of DDGS and lignocellulosic feedstocks, the relevant studies on strain selection, fermentation conditions, targeted metabolism, product nutrition, as well as the growth and health of swine were collected and critically reviewed. This review proposed an approach for the production of easily digestible and highly nutritious swine feed via co-fermentation of DDGS and lignocellulosic feedstocks, which could provide a guide for cleaner swine farming, relieve stress on the increasing demand of high-value swine feed, and finally support the ever-increasing demand of the pork market.

Dimmed gene knockout shortens larval growth and reduces silk yield in the silkworm, Bombyx mori.

The bHLH domain transcription factor, Bombyx mori-derived dimmed (Bmdimm), is directly regulated by the JH-BmMet/BmSRC-BmKr-h1 pathway and plays a key role in regulating the expression of FibH, which codes the main component of silk protein. However, the other roles of Bmdimm in silk protein synthesis remain unclear. Here, we established a Bmdimm knockout (KO) line containing a 7-bp deletion via CRISPR/Cas9 system, which led to the absence of the bHLH domain. The expression level of silk protein genes and silk yield decreased significantly in the Bmdimm KO line. Moreover, knocking out Bmdimm led to shortened larval stages and significant weight loss in larvae and adults. Bmdimm was found to be highly expressed in the silk gland, but it was also expressed in the fat body. The expression level of Bmkr-h1 in the fat body was significantly downregulated in the Bmdimm KO line. Exogenous JHA treatment upregulated Bmkr-h1 and rescued the phenotype of larval growth in the Bmdimm KO line. In conclusion, knocking out Bmdimm led to a shortened larval stage via the inhibition of Bmkr-h1 expression, then reduced silk yield. These findings help to elucidate the regulatory mechanism of fibroin synthesis and larval development in silkworms.

Zearalenone Induces Blood-Testis Barrier Damage through Endoplasmic Reticulum Stress-Mediated Paraptosis of Sertoli Cells in Goats.

Zearalenone (ZEA) is present worldwide as a serious contaminant of food and feed and causes male reproductive toxicity. The implication of paraptosis, which is a nonclassical paradigm of cell death, is unclear in ZEA-induced male reproductive disorders. In this study, the toxic effects of ZEA on the blood-testis barrier (BTB) and the related mechanisms of paraptosis were detected in goats. ZEA exposure, in vivo, caused a significant decrease in spermatozoon quality, the destruction of seminiferous tubules, and damage to the BTB integrity. Furthermore, ZEA exposure to Sertoli cells (SCs) in vitro showed similar dysfunction in structure and barrier function. Importantly, the formation of massive cytoplasmic vacuoles in ZEA-treated SCs corresponded to the highly swollen and dilative endoplasmic reticulum (ER), and paraptosis inhibition significantly alleviated ZEA-induced SC death and vacuolization, which indicated the important contribution of paraptosis in ZEA-induced BTB damage. Meanwhile, the expression of ER stress marker proteins was increased after ZEA treatment but decreased under the inhibition of paraptosis. The vacuole formation and SC death, induced by ZEA, were remarkably blocked by ER stress inhibition. In conclusion, these results facilitate the exploration of the mechanisms of the SC paraptosis involved in ZEA-induced BTB damage in goats.

Isolation and characterization of a novel phage vB_ValP_VA-RY-3 infecting Vibrio alginolyticus.

Vibrio alginolyticus is a common foodborne pathogen existing both in contaminated seafood and the environment and can cause serious mortality in aquaculture facilities. Bacteriophages can be used as an alternative bio-control agent to eliminate and reduce pathogens. In this study, a novel lytic phage, designated vB_ValP_VA-RY-3 (referred to as S1R3Y), was isolated from sewage collected in Dalian, China. The linear double-stranded DNA genome of phage S1R3Y is 40.271 kb, which has a mol% G + C content of 43.98, containing 51 ORFs with a T7-like genomic organization. It shared the closest relationship with phage vB_CsaP_Ss1, but the homology coverage is just 6%. S1R3Y lacks tRNA and no known virulence or lysogenic genes were found. S1R3Y had a burst size of 147 PFU/cell and is stable under different temperatures (4-56 °C) and pH (5.0-7.0). A comparison of its genomic features and phylogenetic analysis revealed that phage S1R3Y is a novel member of the order Caudovirales, family Podoviridae. Our results suggest that phage S1R3Y may represent a potential therapeutic agent against Vibrio alginolyticus.

Evaluation of Biochemical Properties, Antioxidant Activities and Phenolic Content of Two Wild-Grown Berberis Fruits: Berberis nummularia and Berberisatrocarpa.

To evaluate the potential health-promoting benefits of Berberis nummularia and B. atrocarpa fruits, the biochemical properties (nutrition component, mineral substance, organic acids), total phenolic and flavonoid content and antioxidant (DPPH, FRAP, ABTS and ORAC) capacity of ethanol extracts of B. nummularia and B. atrocarpa fruits wild-grown in Xinjiang were analyzed. The results indicated that there were no meaningful differences (p > 0.05) between the ash (1 ± 0.1 and 1 ± 0.0 g/100 g), fiber (16 ± 1.0 and 18 ± 1.4) and carbohydrate (57 ± 1.8 and 56 ± 1.8 g/100 g) content, respectively, in the dry fruits of B. nummularia and B. atrocarpa. The total fat (7 ± 0.4 and 5 ± 0.1 mg/100 g), soluble sugar (23 ± 0.6 and 12 ± 1.4 g/100 g), titratable acidity (18 ± 2.5% and 14 ± 1.3%) content, and energy value (330.86 and 314.41 kcal/100 g) of B. nummularia was significantly higher than that of B. atrocarpa fruits. Both species contain malic acid, acetic acid, tartaric acid, citric acid and fumaric acid, in which, malic acid is the dominant organic acid. The organic acid and mineral components of B. nummularia fruits were significantly higher than that of B. atrocarpa (p < 0.05). The total phenolic and flavonoid content of B. nummularia were 2 ± 0.0 mg GA/g DW and 2 ± 0.0 mg RE/g DW, respectively, which were significantly lower than the total phenolic and flavonoid content of B. atrocarpa (12 ± 0.1 mg GA/g DW and 9 ± 0.0 mg RE/g DW). The antioxidant capacity of B. nummularia (4 ± 0.1 mg Ascorbic acid/g DW for DPPH, 32 ± 0.1 mg Trolox/g DW for FRAP, 80 ± 3.0 mg Trolox/g DW for ABTS and 60 ± 3.6 mg Trolox/g for ORAC was significantly lower than that of B. atrocarpa (12 ± 0.0 mg Ascorbic acid/g DW for DPPH, 645 ± 1.1 mg Trolox/g DW for FRAP, 304 ± 3.0 mg Trolox/g DW for ABTS and 155 ± 2.8 mg Trolox/g for ORAC). B. atrocarpa fruits showed significantly higher antioxidant capacity than that of B. nummularia. The fruits of the two species can be used in food coloring and nutritional supplements, and consumption of the fruits can aid in weight control and reduce blood glucose or cholesterol.

Exosomal lncRNA PCAT1 Promotes Tumor Circulating Cell-Mediated Colorectal Cancer Liver Metastasis by Regulating the Activity of the miR-329-3p/Netrin-1-CD146 Complex.

Objective: This study explored the colorectal cancer exosome lncRNA prostate cancer associated transcript 1- (PCAT1) mediated circulating tumors and the mechanism of cell colorectal cancer liver metastasis. Methods: Exosomes were extracted from the primary colorectal cancer (CRC) cell lines HCT116 and SW480 and cultured with T84 and human umbilical vein endothelial (HUVE) cells. The expression of PCAT1 and miR-329-3p was detected by real-time quantitative polymerase chain reaction (RT-qPCR), the expression of Netrin-1, CD146, and epithelial mesenchymal transition (EMT) related proteins was detected by Western blot, the proliferation activity of T84 cells was detected by cell counting kit 8 (CCK-8), and cell migration was detected by Transwell. The expression of the F-actin signal was detected by immunofluorescence after coculture of exosomes with human umbilical vein endothelial cells (HUVECs). Changes in subcutaneous tumor and liver nodule size after PCAT1 deletion were observed in a mouse model of liver metastasis from rectal cancer. Results: PCAT1 expression was upregulated in primary cell lines and their exosomes. After exosomes were cocultured with colorectal cancer tumor circulating T84 cells, the expression of Netrin-1 and CD146 was upregulated, the expression of miR-329-3p was downregulated, the proliferation and migration ability of T84 cells were enhanced, and EMT occurred. After knocking down PCAT1, the above phenomenon was reversed. Similarly, after exosomes were cocultured with HUVECs, the expression of the F-actin signal increased, and after PCAT1 was knocked down, the F-actin signal also decreased. PCAT1 regulates miR-329-3p/Netrin-1 and affects the biological behavior of T84 and F-actin signal expression in HUVECs. In a mouse model of colorectal cancer liver metastasis, knocking down PCAT1 significantly reduced the nodules formed by liver metastasis in mice. Conclusions: LncRNA PCAT1 derived from colorectal cancer exosomes regulates the activity of the Netrin-1-CD146 complex in circulating tumor cells (CTCs) to promote the occurrence of colorectal cancer EMT and liver metastasis and provides new molecular targets for the treatment of colorectal cancer liver metastasis.

Alterations in the vaginal microbiota of patients with preterm premature rupture of membranes.

Background: Preterm premature rupture of membranes (PPROM) is a common pregnancy complication. Yet, the main cause of PPROM remains poorly understood. In this study, we used 16S rRNA gene sequencing technology to identify the differences in vaginal microbiota between pregnant women with PPROM and those who delivered at term. Methods: Vaginal samples were collected from 48 patients with PPROM and 54 age- and gestational age-matched pregnant women who delivered at term (controls). The vaginal microbiota of the two groups was compared using 16S rRNA gene sequencing of the V3-V4 regions. Results: The vaginal microbial composition of the PPROM group was significantly different from that of the control group. Our results showed that the diversity of vaginal microbiota in patients with PPROM increased compared with controls. The relative abundance of Lactobacillus iners, Gardnerella vaginalis, Prevotella bivia, Ochrobactrum sp., Prevotella timonensis, and Ureaplasma parvum were more abundant in patients with PPROM, while Lactobacillus crispatus and Lactobacillus gasseri were more abundant in controls. Ochrobactrum sp., Prevotella timonensis, and Gardnerella vaginalis, could serve as biomarkers for PPROM. Finally, we proposed several metabolic pathways, including PWY-6339, PWY-6992, and PWY-7295. Conclusion: PPROM is characterized by vaginal microbial dysbiosis. The dysbiotic vaginal microbiota signatures in patients with PPROM include a higher bacterial diversity, decreased autochthonous bacteria, and increased pathogenic bacteria. These results may be beneficial for developing biomarkers for screening and early diagnosis of PPROM and may provide effective preventative treatments.

One-step salting-out extraction of bacteriophage from its infection broth of Acinetobacter baumannii.

Phages as a potential alternative antibiotic for multidrug-resistant bacterial infections are receiving great attention worldwide. However, the traditional separation and purification of phage are cumbersome, time-consuming, costly and inefficient. In this study, phage phiAB9 for multidrug-resistant Acinetobacter baumannii was separated and purified by a simple and cost-saving one-step salting-out extraction (SOE). Several kinds of salts and organic solvents without effect on phage survival were chosen to form the SOE systems, and the composition of SOE systems were optimized according to the phage recovery rate and impurity removal rate. After one-step SOE by an optimal system composed of 18% (w/w) ammonium citrate and 40% (w/w) ethyl acetate, the recovery rate of phage in the middle phase could reach to 90.82%, and most proteins (99.57%), cells (97.98%) and endotoxin (84.08%) were well removed, with a concentration factor of 210 and the purification factors of phage to proteins, cells and endotoxin were 303.64, 133.55 and 5.36, respectively. Comparing with two-step SOE and traditional aqueous two-phase extraction, one-step SOE may be an available method for the separation and purification of phages.