Research advances in the identification of regulatory mechanisms of surfactin production by Bacillus: a review.

Surfactin is a cyclic hexalipopeptide compound, nonribosomal synthesized by representatives of the Bacillus subtilis species complex which includes B. subtilis group and its closely related species, such as B. subtilis subsp subtilis, B. subtilis subsp spizizenii, B. subtilis subsp inaquosorum, B. atrophaeus, B. amyloliquefaciens, B. velezensis (Steinke mSystems 6: e00057, 2021) It functions as a biosurfactant and signaling molecule and has antibacterial, antiviral, antitumor, and plant disease resistance properties. The Bacillus lipopeptides play an important role in agriculture, oil recovery, cosmetics, food processing and pharmaceuticals, but the natural yield of surfactin synthesized by Bacillus is low. This paper reviews the regulatory pathways and mechanisms that affect surfactin synthesis and release, highlighting the regulatory genes involved in the transcription of the srfAA-AD operon. The several ways to enhance surfactin production, such as governing expression of the genes involved in synthesis and regulation of surfactin synthesis and transport, removal of competitive pathways, optimization of media, and fermentation conditions were commented. This review will provide a theoretical platform for the systematic genetic modification of high-yielding strains of surfactin.

Identification of two novel large deletions in FBN1 gene by next-generation sequencing and multiplex ligation-dependent probe amplification.

BACKGROUND: Mutations in fibrillin-1 (FBN1) are known to be associated with Marfan syndrome (MFS), an autosomal dominant connective tissue disorder. Most FBN1 mutations are missense or nonsense mutations. Traditional molecular genetic testing for the FBN1 gene, like Sanger sequencing, may miss disease-causing mutations in the gene's regulatory regions or non-coding sequences, as well as partial or complete gene deletions and duplications. METHODS: Next-generation sequencing, multiplex ligation-dependent probe amplification and gap PCR were conducted on two MFS patients to screen for disease-causing mutations. RESULTS: We identified two large deletions in FBN1 from two MFS patients. One patient had a 0.23 Mb deletion (NC_000015.9:g.48550506_48779360del) including 5'UTR-exon6 of FBN1. The other patient harbored a 1416 bp deletion (NC_000015.9:g.48410869_48412284del) affecting the last exon, exon 66, of the FBN1 gene. CONCLUSION: Our results expanded the number of large FBN1 deletions and highlighted the importance of screening for large deletions in FBN1 in clinical genetic testing, especially for those with the classic MFS phenotype.

First Report of Gilbertella persicaria causing Postharvest Soft Rot of Pyrus pyrifolia 'Housui' in China.

Postharvest diseases lead to substantial economic losses to the pear industry (Xu et al. 2021). In August 2022 and 2023, 'Housui' pears (Pyrus pyrifolia) with no visible wounds were harvested from Baoying county, Jiangsu Province, China and stored at 20°C with 85% relative humidity. Approximately 8% of pear fruits showed soft rot after 15 days of storage. The margin area of rot tissue was aseptically incubated on PDA medium at 25°C. Mycelial tips were transferred to new PDA after 24 h. Five fungal isolates were obtained after isolation and identification, including Alternaria sp., Botryosphaeria sp., Diaporthe sp., Fusarium sp. and Gilbertella sp. For each isolate, pathogenicity tests were confirmed three times by placing 10 µL of spore suspension (106 spores/mL) on three 'Housui' pear fruits superficially wounded with sterile toothpicks, and sterile distilled water served as controls. Lesions caused by Gilbertella sp. were distinctly observed after incubating at 20°C for 24h, and controls have no symptom. The lesions expanded to large brown spots with smelling of alcohol after 48 h, similar to natural disease symptom. The colony of Gilbertella sp. was initially white and rapidly turned gray, generating large amounts of black sporangia. -Sporangia were firstly white, then turn black, globose to dorsoventrally flattened, 70.22 to 131.58 × 75 to 135.17 µm, average 93.19 × 106.54 µm (n = 50), borne erect or nodding, breaking into two equal pieces. Sporangiophores were hyaline, 11.17 to 34.57 µm wide, average 19.67 µm (n = 50). Columellae were hyaline, pyriform or obovoid to cylindrical, with a distinct basal collar, 32.37 to 102.84 × 23.62 to 68.68 µm, average 60.06 × 40.07 µm (n = 50). Sporangiospores were single celled, mostly ellipsoid, 5.76 to 11.49 × 3.89 to 6.18 µm, average 8.68 × 5.08 µm (n = 100), attaching with 4-5 hyaline appendages at the ends. Chlamydospores were solitary or in short chain, cylindrical or oval. Zygospore was not observed. The isolate was morphologically identified as G. persicaria (Benny 1991). Molecular identification was performed by PCR amplification, sequencing and phylogenetic analysis of the internal transcribed spacer region of rDNA (ITS), partial 28S rDNA large subunit (LSU), and actin-1 (ACT-1) gene using primer pairs ITS1/ITS4, LR0R/LR5 and Gil_ACT_F/Gil_ACT_R (Zhang et al. 2020). The ITS (OP897009), LSU (OR794326), and ACT-1 (OR805109) sequences revealed 99.85%, 99.30% and 100% sequence identity to nucleotide sequences of G. persicaria from NCBI (ON875318, OP243274, and AJ287159). Phylogenetic analysis based on the maximum likelihood method grouped the isolate with other G. persicaria strains. Pathogenicity of the isolate was performed on wounded and non-wounded fruits. Wounded fruits severely rot after 48 h, and no non-wounded fruit rot after 5 days. Therefore, wound was required for the infection of G. persicaria. The pathogen was consistently re-isolated and purified from the inoculated pears, morphologically identified as G. persicaria, fulfilling Koch's postulates. Fruit rot caused by G. persicaria has been reported on peach, tomato, apricot, plum, apple, dragon fruit, papaya and eggplant, as well as Pyrus communis (Mehrotra 1964; Ginting et al. 1996; Cruz-Lachica et al. 2021). This is the first report of G. persicaria infection on 'Housui' pears in China. This disease is a potential threat to 'Housui' pear storage. The confirmation of this soft rot pathogen provides a foundation for pear postharvest disease prevention.

Assessment of Auricularia cornea var. Li. polysaccharides potential to improve hepatic, antioxidation and intestinal microecology in rats with non-alcoholic fatty liver disease.

Non-alcoholic fatty acid liver disease (NAFLD) is a reputed global health concern, affecting children and young adults. Accumulating evidence suggests that edible fungi polysaccharides have the potential to relieve NAFLD. Our previous study found that Auricularia cornea var. Li. polysaccharides (ACP) could improve immune by regulating gut microbiota. However, its NAFLD-alleviating potentials have been scarcely reported. This study analyzed the protective effects of Auricularia cornea var. Li. polysaccharides on high-fat diet (HFD)-induced NAFLD and mechanistic actions. We first analyzed the histology and hepatic lipid profile of animals to evaluate this variant's ameliorating effects on NAFLD. Then, antioxidant and anti-inflammatory potentials of ACP were studied. Finally, we explored changes in the gut microbiome diversity for mechanistic insights from the gut-liver region. Results showed that supplementation with ACP substantially reduced homeostasis model assessment-insulin resistance (HOMA-IR), body fat, liver index rates and weight gain (p < 0.05). This variant also improved HDL-C levels while decreasing triglyceride (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C) levels which were initially triggered by HFD. ACP mediation also decreased the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels considerably with H&E technique indicating that it can reduce liver lipid accumulation, thus lowering liver damages risks (p < 0.05). The antioxidant potentials of ACP were also demonstrated as it decreased the hepatic levels of malondialdehyde (MDA) and increased the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX). Proinflammatory markers like IL-6, IL-1ß and TNF-α concentrations were decreased by ACP supplementation, accompanied with increased IL-4 levels. Finally, ACP supplementation regulated the intestinal microbiota to near normal patterns. In all, ACP protects HFD-induced NAFLD by improving liver characteristics and regulating colonic flora composition, our findings assert that ACP can be a promising strategy in NAFLD therapy.

Ectopic Expression of Gastrodia Antifungal Protein in Rice Enhances Resistance to Rice Sheath Blight Disease.

Sheath blight (ShB) disease, caused by Rhizoctonia solani Kühn, is one of the most serious rice diseases. Rice breeding against ShB has been severely hindered because no major resistance genes or germplasms are available in rice. Here, we report that introduction of Gastrodia antifungal protein (GAFP) genes from Gastrodia elata B1 into rice significantly enhances resistance to rice ShB. Four GAFP genes were cloned from G. elata B1, and all displayed a strong ability to inhibit R. solani growth in plate assays. Two versions, with or without a signal peptide, for each of the four GAFP genes were introduced into XD3 and R6547 rice cultivars, and all transgenic lines displayed stronger ShB resistance than the corresponding wild-type control in both greenhouse and field conditions. Importantly, GAFP2 showed the highest ShB resistance; GAFPs with and without its signal peptide showed no significant differences in enhancing ShB resistance. We also evaluated the agronomic traits of these transgenic rice and found that ectopic expression of GAFPs in rice at appropriate levels did not affect agronomic traits other than enhancing ShB resistance. Together, these results indicate that GAFP genes, especially GAFP2, have great potential in rice breeding against ShB disease.

Nanosheet-Facilitated Spray Delivery of dsRNAs Represents a Potential Tool to Control Rhizoctonia solani Infection.

Rhizoctonia solani is one of the important pathogenic fungi causing several serious crop diseases, such as maize and rice sheath blight. Current methods used to control the disease mainly depend on spraying fungicides because there is no immunity or high resistance available in crops. Spraying double-strand RNA (dsRNA) for induced-gene silencing (SIGS) is a new potentially sustainable and environmentally friendly tool to control plant diseases. Here, we found that fluorescein-labelled EGFP-dsRNA could be absorbed by R. solani in co-incubation. Furthermore, three dsRNAs, each targeting one of pathogenicity-related genes, RsPG1, RsCATA, and RsCRZ1, significantly downregulated the transcript levels of the target genes after co-incubation, leading to a significant reduction in the pathogenicity of the fungus. Only the spray of RsCRZ1 dsRNA, but not RsPG1 or RsCATA dsRNA, affected fungal sclerotium formation. dsRNA stability on leaf surfaces and its efficiency in entering leaf cells were significantly improved when dsRNAs were loaded on layered double hydroxide (LDH) nanosheets. Notably, the RsCRZ1-dsRNA-LDH approach showed stronger and more lasting effects than using RsCRZ1-dsRNA alone in controlling pathogen development. Together, this study provides a new potential method to control crop diseases caused by R. solani.

Regulatory effects of Auricularia cornea var. Li. polysaccharides on immune system and gut microbiota in cyclophosphamide-induced mice.

The immuno-regulating potential of edible fungus polysaccharides has gained more and more attention. However, there is little information about the study of Auricularia cornea var. Li. polysaccharides regulating immunomodulatory activity. The objective of this work to analyze the immunomodulatory activity and the mechanism of A. cornea var. Li. polysaccharides supplementation in an immunosuppressed mice model induced by cyclophosphamide. The effects of A. cornea var. Li. polysaccharides on immune system including immune organ indices, immunoglobulin contents, and inflammation cytokines in immunosuppressed mice were determined. In addition, the regulatory effects of A. cornea var. Li. polysaccharides on the gut microbiota and their metabolites were analyzed. Results showed that A. cornea var. Li. polysaccharides significantly elevated immune organ indexes, remarkably enhanced the levels of immunoglobulin A (IgA), IgG and IgM in serum and secretory IgA (sIgA) in the intestinal mucosa, conspicuously stimulated the levels of tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), IL-4, and IL-10 in the serum. A. cornea var. Li. polysaccharides also could restore gut microbiota to the pattern that is similar with that of the control group with increase of the relative abundances of short-chain fatty acids (SCFAs)-producing bacteria. Furthermore, the content of SCFAs were increased after A. cornea var. Li. polysaccharides supplementation. This study provides useful information for applications of A. cornea var. Li. polysaccharides in immune-regulated foods and medicine.

Correlations among population genetic structure, geographic origin, growth rate, and fungicide resistance of Rhizoctonia solani AG 1-IA, the pathogen of rice sheath blight.

Rhizoctonia solani AG1-IA is a necrotrophic fungus that causes rice sheath blight and results in severe yield and quality reductions in rice worldwide. Differences of genetic structure and fungicide sensitivity of the pathogen have significant effects on the severity and control effect of this disease in the field. To determine correlations among population genetic structure, geographic origin, growth rate, and fungicide resistance of the pathogen, 293 strains of R. solani were isolated from diseased rice collected from 13 cities of Jiangsu Province and five regions of China. Simple sequence repeat (SSR) molecular marker technology was used to analyze the genetic diversity of these strains, and a total of 74 bands were amplified by nine pairs of primers. Population genetic structure analysis showed that strains from Central China and northern Jiangsu had the highest Nei's gene diversity index and Shannon diversity index. The vast majority of strains grew fast with colony diameters of more than 60.0 mm cultured at 28 °C for 36 h. The half-maximal effective concentration (EC50) of them to tebuconazole, thifluzamide, and propiconazole varied ∼16.2-, 3.8-, and 7.5-fold. However, the genetic diversity of R. solani had no significant correlation with their geographic origin, growth rate or fungicide sensitivity.

Transcriptome profiling and co-expression network analysis of lncRNAs and mRNAs in colorectal cancer by RNA sequencing.

BACKGROUND: Long non-coding RNAs (lncRNAs) are widely involved in the pathogenesis of cancers. However, biological roles of lncRNAs in occurrence and progression of colorectal cancer (CRC) remain unclear. The current study aimed to evaluate the expression pattern of lncRNAs and messenger RNAs (mRNAs). METHODS: RNA sequencing (RNA-Seq) in CRC tissues and adjacent normal tissues from 6 CRC patients was performed and functional lncRNA-mRNA co-expression network was constructed afterwards. Gene enrichment analysis was demonstrated using DAVID 6.8 tool. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to validate the expression pattern of differentially expressed lncRNAs. Pearson correlation analysis was applied to evaluate the relationships between selected lncRNAs and mRNAs. RESULTS: One thousand seven hundred and sixteenth differentially expressed mRNAs and 311 differentially expressed lncRNAs were screened out. Among these, 568 mRNAs were up-regulated while 1148 mRNAs down-regulated, similarly 125 lncRNAs were up-regulated and 186 lncRNAs down-regulated. In addition, 1448 lncRNA-mRNA co-expression pairs were screened out from 940,905 candidate lncRNA-mRNA pairs. Gene enrichment analysis revealed that these lncRNA-related mRNAs are associated with cell adhesion, collagen adhesion, cell differentiation, and mainly enriched in ECM-receptor interaction and PI3K-Akt signaling pathways. Finally, RT-qPCR results verified the expression pattern of lncRNAs, as well as the relationships between lncRNAs and mRNAs in 60 pairs of CRC tissues. CONCLUSIONS: In conclusion, these results of the RNA-seq and bioinformatic analysis strongly suggested that the dysregulation of lncRNA is involved in the complicated process of CRC development, and providing important insight regarding the lncRNAs involved in CRC.

RBCK1 promotes hepatocellular carcinoma metastasis and growth by stabilizing RNF31.

RNF31 (HOIP), RBCK1 (HOIL-1L), and SHARPIN are subunits of the linear ubiquitin chain assembly complex. Their function and specific molecular mechanisms in hepatocellular carcinoma (HCC) have not been reported previously. Here, we investigated the role of RNF31 and RBCK1 in HCC. We showed that RNF31 and RBCK1 were overexpressed in HCC and that upregulation of RNF31 and RBCK1 indicated poor clinical outcomes in patients with HCC. RNF31 overexpression was significantly associated with more satellite foci and vascular invasion in patients with HCC. Additionally, RBCK1 expression correlated positively with RNF31 expression in HCC tissues. Functionally, RBCK1 and RNF31 promote the metastasis and growth of HCC cells. Moreover, the RNF31 inhibitor gliotoxin inhibited the malignant behavior of HCC cells. Mechanistically, RBCK1 interacted with RNF31 and repressed its ubiquitination and proteasomal degradation. In summary, the present study revealed an oncogenic role and regulatory relationship between RBCK1 and RNF31 in facilitating proliferation and metastasis in HCC, suggesting that they are potential prognostic markers and therapeutic targets for HCC.

Long non-coding RNA muskelin 1 antisense RNA as a potential therapeutic target in hepatocellular carcinoma treatment.

Long non-coding RNAs are essential to hepatocellular carcinoma (HCC) development, progression, and incidence of drug resistance. However, the biological significance of long non-coding RNA muskelin 1 antisense RNA (MKLN1-AS) remains poorly characterized. In this study, we observed noticeable increased levels of MKLN1-AS in HCC tissues. This upregulation of MKLN1-AS was clinically associated with vascular invasion and decreased disease-free survival and overall survival of patients with HCC. Functionally, MKLN1-AS-knockdown dramatically suppressed the metastasis and growth of HCC cells in vitro and in vivo. Additionally, the knockdown of MKLN1-AS augmented the pro-apoptosis effect of lenvatinib. Taken together, our findings indicate that MKLN1-AS may be exploited as a potential prognostic predictor and therapeutic target for HCC treatment.

Suppressing chlorophyll degradation by silencing OsNYC3 improves rice resistance to Rhizoctonia solani, the causal agent of sheath blight.

Necrotrophic fungus Rhizoctonia solani Kühn (R. solani) causes serious diseases in many crops worldwide, including rice and maize sheath blight (ShB). Crop resistance to the fungus is a quantitative trait and resistance mechanism remains largely unknown, severely hindering the progress on developing resistant varieties. In this study, we found that resistant variety YSBR1 has apparently stronger ability to suppress the expansion of R. solani than susceptible Lemont in both field and growth chamber conditions. Comparison of transcriptomic profiles shows that the photosynthetic system including chlorophyll biosynthesis is highly suppressed by R. solani in Lemont but weakly in YSBR1. YSBR1 shows higher chlorophyll content than that of Lemont, and inducing chlorophyll degradation by dark treatment significantly reduces its resistance. Furthermore, three rice mutants and one maize mutant that carry impaired chlorophyll biosynthesis all display enhanced susceptibility to R. solani. Overexpression of OsNYC3, a chlorophyll degradation gene apparently induced expression by R. solani infection, significantly enhanced ShB susceptibility in a high-yield ShB-susceptible variety '9522'. However, silencing its transcription apparently improves ShB resistance without compromising agronomic traits or yield in field tests. Interestingly, altering chlorophyll content does not affect rice resistance to blight and blast diseases, caused by biotrophic and hemi-biotrophic pathogens, respectively. Our study reveals that chlorophyll plays an important role in ShB resistance and suppressing chlorophyll degradation induced by R. solani infection apparently improves rice ShB resistance. This discovery provides a novel target for developing resistant crop to necrotrophic fungus R. solani.

Dimethyl itaconate inhibits LPS­induced microglia inflammation and inflammasome­mediated pyroptosis via inducing autophagy and regulating the Nrf­2/HO­1 signaling pathway.

The endogenous metabolite itaconate and its cell­permeable derivative dimethyl itaconate (DI) have been identified as anti­inflammatory regulators of macrophages; however, their contribution to inflammasome­mediated pyroptosis remains unknown. The present study examined the molecular mechanism of DI on NLR family pyrin domain­containing 3 (NLRP3) inflammasome assembly and NLRP3 inflammasome­dependent pyroptosis in microglia. Lipopolysaccharide (LPS) and ATP were used to induce microglia pyroptosis in vitro; this process was confirmed by TUNEL assay, lactate dehydrogenase (LDH) detection and gasdermin D (GSDMD) expression analysis. The regulation of microglia polarization and inflammatory cytokine expression was assessed by immunofluorescence assays and ELISA. To investigate the associated mechanism of action, the expression levels of the nuclear factor erythroid 2­related factor 2 (Nrf­2)/heme oxygenase­1 (HO­1) pathway proteins were analyzed by western blotting. Finally, the regulatory effect of DI on autophagy and its association with inflammation was determined by western blotting. The present study demonstrated that DI administration inhibited NLRP3 assembly, LDH release and GSDMD cleavage. Cotreatment of DI with LPS and ATP facilitated the transition from M1 to M2, reduced inflammatory mediator expression and impeded NF­κB phosphorylation. In addition, DI effectively reduced reactive oxygen species production through the Nrf­2/HO­1 pathway. Moreover, DI induced cellular autophagy, whereas inhibition of autophagy with 3­methyladenine markedly reversed its inhibitory effect on NLRP3­dependent pyroptosis. Taken together, the present study suggested that DI participated in the Nrf­2/HO­1 pathway and served a key role in microglia inflammation and NLRP3 inflammasome­mediated pyroptosis via induction of autophagy.

The dynamics of N6-methyladenine RNA modification in interactions between rice and plant viruses.

BACKGROUND: N6-methyladenosine (m6A) is the most common RNA modification in eukaryotes and has been implicated as a novel epigenetic marker that is involved in various biological processes. The pattern and functional dissection of m6A in the regulation of several major human viral diseases have already been reported. However, the patterns and functions of m6A distribution in plant disease bursting remain largely unknown. RESULTS: We analyse the high-quality m6A methylomes in rice plants infected with two devastating viruses. We find that the m6A methylation is mainly associated with genes that are not actively expressed in virus-infected rice plants. We also detect different m6A peak distributions on the same gene, which may contribute to different antiviral modes between rice stripe virus or rice black-stripe dwarf virus infection. Interestingly, we observe increased levels of m6A methylation in rice plant response to virus infection. Several antiviral pathway-related genes, such as RNA silencing-, resistance-, and fundamental antiviral phytohormone metabolic-related genes, are also m6A methylated. The level of m6A methylation is tightly associated with its relative expression levels. CONCLUSIONS: We revealed the dynamics of m6A modification during the interaction between rice and viruses, which may act as a main regulatory strategy in gene expression. Our investigations highlight the significance of m6A modifications in interactions between plant and viruses, especially in regulating the expression of genes involved in key pathways.

ChIP-seq Analysis of the Global Regulator Vfr Reveals Novel Insights Into the Biocontrol Agent Pseudomonas protegens FD6.

Many Pseudomonas protegens strains produce the antibiotics pyoluteorin (PLT) and 2,4-diacetylphloroglucinol (2,4-DAPG), both of which have antimicrobial properties. The biosynthesis of these metabolites is typically controlled by multiple regulatory factors. Virulence factor regulator (Vfr) is a multifunctional DNA-binding regulator that modulates 2,4-DAPG biosynthesis in P. protegens FD6. However, the mechanism by which Vfr regulates this process remains unclear. In the present study, chromatin immunoprecipitation of FLAG-tagged Vfr and nucleotide sequencing analysis were used to identify 847 putative Vfr binding sites in P. protegens FD6. The consensus P. protegens Vfr binding site predicted from nucleotide sequence alignment is TCACA. The qPCR data showed that Vfr positively regulates the expression of phlF and phlG, and the expression of these genes was characterized in detail. The purified recombinant Vfr bound to an approximately 240-bp fragment within the phlF and phlG upstream regions that harbor putative Vfr consensus sequences. Using electrophoretic mobility shift assays, we localized Vfr binding to a 25-bp fragment that contains part of the Vfr binding region. Vfr binding was eliminated by mutating the TACG and CACA sequences in phlF and phlG, respectively. Taken together, our results show that Vfr directly regulates the expression of the 2,4-DAPG operon by binding to the upstream regions of both the phlF and phlG genes. However, unlike other Vfr-targeted genes, Vfr binding to P. protegens FD6 does not require an intact binding consensus motif. Furthermore, we demonstrated that vfr expression is autoregulated in this bacterium. These results provide novel insights into the regulatory role of Vfr in the biocontrol agent P. protegens.

A synonymous mutation in exon 39 of FBN1 causes exon skipping leading to Marfan syndrome.

Marfan syndrome is a heritable autosomal-dominant connective tissue disorder and it was typically caused by mutations in FBN1. However, the synonymous mutation was seldom recorded to be related to Marfan syndrome. Hereon, Multiplex ligation-dependent probe amplification failed to detect a copy number variant involving FBN1 but a synonymous mutation c.4773A > G (p.Gly1591Gly) was identified by NGS in exon 39. RNA was extracted from patient's aortic tissue and reverse polymerase chain reaction demonstrated the presence of a shortened mRNA transcript. Results of minigene models indicated that c.4773A > G was bona fide responsibility for the aberrant splicing pattern, and artificial mutations of c.4773A > C and c.4773A > T also gave rise to fragments with exon 39 entire skipped. Together, the novel synonymous mutations in c.4773 position (A > G, C, T), middle of exon 39 of FBN1 gene, was found to be associated with Marfan syndrome by altering the splicing pattern of pre-mRNA.

RNA m6 A methylation regulates sorafenib resistance in liver cancer through FOXO3-mediated autophagy.

N6-methyladenosine (m6 A) is an abundant nucleotide modification in mRNA, known to regulate mRNA stability, splicing, and translation, but it is unclear whether it is also has a physiological role in the intratumoral microenvironment and cancer drug resistance. Here, we find that METTL3, a primary m6 A methyltransferase, is significantly down-regulated in human sorafenib-resistant hepatocellular carcinoma (HCC). Depletion of METTL3 under hypoxia promotes sorafenib resistance and expression of angiogenesis genes in cultured HCC cells and activates autophagy-associated pathways. Mechanistically, we have identified FOXO3 as a key downstream target of METTL3, with m6 A modification of the FOXO3 mRNA 3'-untranslated region increasing its stability through a YTHDF1-dependent mechanism. Analysis of clinical samples furthermore showed that METTL3 and FOXO3 levels are tightly correlated in HCC patients. In mouse xenograft models, METTL3 depletion significantly enhances sorafenib resistance of HCC by abolishing the identified METTL3-mediated FOXO3 mRNA stabilization, and overexpression of FOXO3 restores m6 A-dependent sorafenib sensitivity. Collectively, our work reveals a critical function for METTL3-mediated m6 A modification in the hypoxic tumor microenvironment and identifies FOXO3 as an important target of m6 A modification in the resistance of HCC to sorafenib therapy.

PRSS1 genotype is associated with prognosis in patients with pancreatic ductal adenocarcinoma.

The prognostic value of the genotype of the PRSS1 gene in patients with pancreatic ductal adenocarcinoma (PDAC) remains poorly understood. The aim of the present study was to evaluate the association between the PRSS1 genotype and clinicopathological characteristics of patients with PDAC, as well as to explore the prognostic significance of the PRSS1 genotype in patients with PDAC. A total of 124 patients with PDAC patients were included in the current study and the PRSS1 genotype of the enrolled patients was determined by the polymerase chain reaction. Associations between the PRSS1 genotype and clinicopathological characteristics were subsequently analyzed using the Chi-square test. The impact of the PRSS1 genotype on patient prognosis was assessed using the Kaplan-Meier method, and predictive factors of overall survival (OS) time were analyzed by Cox regression. A total of 56 patients with PDAC (45.16%) had the T/C PRSS1 genotype, which was associated with large tumor sizes (P=0.027) and higher tumor node metastasis (TNM) stages (P=0.041). Following a median follow-up of 19 months, the T/C genotype of PRSS1 genotype was associated with a shorter OS time (P=0.037) compared with the C/C or T/T PRSS1 genotypes. Univariate and multivariate analyses revealed that PRSS1 genotype was identified to be an independent prognostic factor for the OS time of patients with PDAC. The results obtained in the current study suggested that the PRSS1 genotype, as well as factors such as the serum level of carbohydrate antigen 19-9 and the TNM stage, may act as independent prognostic factors for the OS time of patients with PDAC.

FolVps9, a Guanine Nucleotide Exchange Factor for FolVps21, Is Essential for Fungal Development and Pathogenicity in Fusarium oxysporum f. sp. lycopersici.

The soil-borne, asexual fungus Fusarium oxysporum f.sp. lycopersici (Fol) is the causal agent of tomato wilt disease. Autophagy plays a crucial role in the development and virulence of Fol. The Fol endosomal system is highly dynamic and has been shown to be associated with conidiogenesis and pathogenicity. Rab GTPases and the regulators are highly conserved in regulating autophagy and endocytosis in most eukaryotes. Identification and characterization of additional Rab regulators in fungal pathogens should facilitate the understanding of the autophagy and endocytosis in different filamentous fungi. Here, we have identified and characterized the yeast VPS9 homolog FolVPS9 in Fol. Targeted gene deletion showed that FolVPS9 is important for growth, conidiation and virulence in Fol. Cytological examination revealed that FolVps9 co-localized with FolVps21 (a marker of early endosome) and played a critical role in endocytosis and autophagosome degradation. Pull-down assays showed that FolVps9 interacted with FolVps21, which was also important for development and plant infection in Fol. Yeast two-hybrid, bimolecular fluorescence complementation and co-immunoprecipitation assays revealed that FolVps9 specifically interacts with the GDP-bound form of FolVps21. Furthermore, a constitutively active form of FolVps21 (Q72L) was able to rescue defects of ΔFolvps9 and ΔFolvps21 mutants. In summary, our study provides solid evidence that FolVps9 acts as a FolVps21 guanine nucleotide exchange factor (GEFs) to modulate endocytosis and autophagy, thereby controlling vegetative growth, asexual development and pathogenicity in Fol.

A value-added multistage utilization process for the gradient-recovery tin, iron and preparing composite phase change materials (C-PCMs) from tailings.

Tin-, iron-bearing tailing is a typically hazardous solid waste in China, which contains plenty of valuable tin, iron elements and is not utilized effectively. In this study, a multistage utilization process was put forward to get the utmost out of the valuable elements (tin and iron) from the tailings, and a gradient-recovery method with three procedures was demonstrated: (1) An activated roasting followed by magnetic separation process was conducted under CO-CO2 atmosphere, tin and iron were efficiently separated during magnetic separation process, and 90.8 wt% iron was enriched in magnetic materials while tin entered into non-magnetic materials; (2) The tin-enriched non-magnetic materials were briquetted with CaCl2 and anthracite and roasted, then tin-rich dusts were collected during the chloridizing roasting process; (3) The roasted briquettes were infiltrated in melting NaNO3 to prepare NaNO3/C-PCMs by a infiltration method. Three kinds of products were obtained from the tailings by the novel process: magnetic concentrates containing 64.53 wt.% TFe, tin-rich dusts containg 52.4 wt.% TSn and NaNO3/C-PCMs for high temperature heat storage. Such a comprehensive and clean utilization method for tin-, iron-bearing tailings produced no secondary hazardous solid wastes, and had great potential for practical application.