The novel oncogenic factor TET3 combines with AHR to promote thyroid cancer lymphangiogenesis via the HIF-1α/VEGF signaling pathway.

BACKGROUND: Lymphangiogenesis has been reported to play crucial roles in the metastasis of thyroid cancer (THCA), but despite the significant research on lymphangiogenesis in THCA, the precise regulatory mechanism remains unclear. METHODS: Public databases including the Cancer Genome Atlas (TCGA), TIMER, and UALCAN were used to analyze and visualize the expression of TET3 and AHR in THCA, and the correlation between these molecules were used by TIMER. Additionally, RT-PCR and Western Blot were performed to determine the mRNA and protein expression of related proteins. Plate colony formation, wound healing, cell cycle, apoptosis, angiogenesis and transwell assay were used to examine the ability of proliferation, movement, lymphangiogenesis, migration and invasion of THCA cells. RESULTS: Analysis of the TCGA database revealed higher expression levels of TET3 and AHR in tumor tissue compared to normal tissue in THCA. Additionally, a strong correlation was observed between TET3 and AHR. UALCAN database demonstrated that high expression of TET3 and AHR was associated with advanced THCA TNM stages in THCA patients. Furthermore, TET3 activation accelerated THCA cell proliferation by inducing G2/M phase arrest and suppressing apoptosis, while AHR inactivation reduced THCA cell proliferation by decreasing G2/M phase arrest and promoting apoptosis in vitro. Notably, both TET3 and AHR significantly enhanced THCA cell lymphangiogenesis, migration and invasion. Moreover, TET3 activation and AHR inactivation regulated HIF-1α/VEGF signaling pathway, which ultimately, blocked the HIF-1α/VEGF in THCA cells and impaired their movement, migration and invasion abilities. CONCLUSIONS: The combined action of TET3 and AHR to promote lymphangiogenesis in THCA through the HIF-1α/VEGF signaling pathway, and targeting them might provide a potential treatment strategy for THCA.

The potential markers of NK-92 associated to cytotoxicity against K562 cells.

Markers associated to NK cytolytic activity are in a great need to regulate NK cell immunotherapy products. We assume that biomarkers which response to cytolysis will change their transcription, expression or secretion. To find NK-92 indicator to cytolytic activity, we have evaluated the potential markers by quantifying the expression of well-known cytotoxicity functional molecules (cytokine IFN-γ, Granzyme B, perforin, CD69 and CD107a), and explored candidate markers by a sweeping transcription picture of NK-92 using a direct cytolysis model (incubation with K562). We found that IFN-γ secretion was highly correlated to cytotoxicity of NK-92, neither Granzyme B, perforin secretion, nor CD69, CD107a positive population were upregulated by K562 stimulation. RNAseq revealed 432 genes expression changed during cytolysis, several genes (BIRC3, CSF2, VCAM1 and TNFRSF9) mRNA expression were validated by real time RT-PCR under K562 being killed or protected from being killed conditions. Results suggested IFN-γ secretion, BIRC3 and TNFRSF9 transcription in NK-92 were responsive to K562 cytolysis. In a word, our results confirmed one marker and reveal an array of novel candidate markers associated with NK-92 cytotoxicity. Further studies are greatly needed to determine the roles these new makers play in NK-92 cytolysis process.

Diagnostic and prognostic value of serum thioredoxin and DJ-1 in non-small cell lung carcinoma patients.

In response to reactive oxygen species (ROS), thioredoxin and DJ-1 are upregulated to counteract the detrimental effect of ROS under normal condition. However, cancer cells can take advantage of thioredoxin and DJ-1 against ROS-induced cell damage. In several human cancer types, thioredoxin and DJ-1 were found to be overexpressed. The present study aimed to explore the serum levels of thioredoxin and DJ-1 in non-small cell lung cancer (NSCLC) patients and its relationship to the diagnosis and prognosis of this particular malignancy. Sera from 134 NSCLC patients and 168 healthy controls were obtained. Using the enzyme-linked immunosorbent assay (ELISA) method, the levels of serum thioredoxin and DJ-1 were measured and correlated to the clinicopathological characteristics of NSCLC patients. The diagnostic and prognostic significance of the biomarkers were evaluated by using receiver operating curve (ROC), Kaplan-Meier curve, and log-rank analyses and the Cox proportional hazard model, respectively. Serum thioredoxin and DJ-1 levels were significantly higher in the NSCLC patients than that in the controls (23.5 ± 6.57 vs. 13.8 ± 2.49 and 7.11 ± 2.02 vs. 5.18 ± 1.26, respectively). NSCLC patients at later stage cancer showed significantly higher levels of serum thioredoxin and DJ-1 than those at the early stages (P < 0.01 and P < 0.05, respectively). Multivariate logistic regression analysis showed that high serum thioredoxin level was an independent risk factor for lymph nodal metastases and distant metastases (OR = 2.18, 95 % CI 1.26-3.41 and OR = 3.68, 95 % CI 2.16-5.33, respectively). In addition, an increase in the serum DJ-1 level was also identified as an independent risk factor for nodal metastases (OR = 1.37, 95 % CI 1.11-3.04). For predicting the development of NSCLC, ROC/area under the curve (AUC) analysis for thioredoxin indicated an AUC of 0.80 (sensitivity 0.62, specificity 0.92), and ROC/AUC analysis for DJ-1 showed an AUC of 0.78 (sensitivity 0.66, specificity 0.89). NSCLC patients with high serum thioredoxin and DJ-1 levels had lower survival rates than those with low levels, and multivariate analyses for overall survival revealed that high serum thioredoxin levels served as an independent prognostic factor for NSCLC (HR = 2.07, 95 % CI 1.19-3.48). Serum levels of thioredoxin and DJ-1 were significantly higher in NSCLC patients; therefore, these may be utilized as novel diagnostic and prognostic biomarkers for NSCLC.

Expression and characterization of hepatitis E virus-like particles and non-virus-like particles from insect cells.

The hepatitis E virus (HEV) capsid antigen expressed in insect cell has been proposed as a candidate subunit vaccine for the prevention of hepatitis E. However, the expression and purification of HEV virus-like particles (VLPs) from insect cells have not been explored. We aimed to optimize the procedure to obtain HEV VLPs. In this study, two conformations of the HEV capsid proteins were expressed in insect cells, VLPs and non-VLPs, and they were purified separately. The physicochemical properties and the humoral immune responses induced by the two forms were analyzed and compared. We found that HEV VLPs were more immunogenic in mice than HEV non-VLPs. Therefore, we optimized the conditions that yielded high VLPs expression in insect cell cultures and developed an efficient purification method. The results suggest that the distinction and isolation of VLPs from non-VLPs are essential to generate a more immunogenic vaccine.

Pseudoxanthomonas wuyuanensis sp. nov., isolated from saline-alkali soil.

A bacterium, designated XC21-2(T), was isolated from a saline-alkaline soil sample from China. Cells were Gram-stain-negative, rod-shaped and motile and grew optimally at 35-37 °C, pH 6.0-7.0 and in the presence of 0.5% (w/v) NaCl. Growth occurred in the range pH 5.5-9.0 and in the presence of up to 4 % (w/v) NaCl. The major cellular fatty acids were iso-C15 : 0, iso-C16 : 0 and iso-C17 : 1ω9c. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an uncharacterized amino-group-containing polar lipid. The major quinone was ubiquinone 8 (Q-8) and the G+C content of the genomic DNA was 66.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain XC21-2(T) formed a tight phylogenetic lineage with Pseudoxanthomonas dokdonensis KCTC 12543(T) within the genus Pseudoxanthomonas and was most closely related to P. dokdonensis KCTC 12543(T) and P. mexicana ATCC 700993(T), with 97.9 and 97.5 % 16S rRNA gene sequence similarity, respectively. On the basis of the unique physiological profile of the isolate and its phylogenetic divergence from known species, strain XC21-2(T) represents a novel species within the genus Pseudoxanthomonas, for which the name Pseudoxanthomonas wuyuanensis sp. nov. is proposed. The type strain is XC21-2(T) ( = CGMCC 1.10978(T) = KCTC 23877(T)).

Cloning and Functional Characterization of 2-C-methyl-D-erythritol-4-phosphate cytidylyltransferase (LiMCT) Gene in Oriental Lily (Lilium 'Sorbonne').

2-C-methyl-D-erythritol-phosphate cytidylyltransferase (MCT) is a key enzyme in the MEP pathway of monoterpene synthesis, catalyzing the generation of 4- (5'-pyrophosphate cytidine)-2-C-methyl-D-erythritol from 2-C-methyl-D-erythritol-4-phosphate. We used homologous cloning strategy to clone gene, LiMCT, in the MEP pathway that may be involved in the regulation of floral fragrance synthesis in the Lilium oriental hybrid 'Sorbonne.' The full-length ORF sequence was 837 bp, encoding 278 amino acids. Bioinformatics analysis showed that the relative molecular weight of LiMCT protein is 68.56 kD and the isoelectric point (pI) is 5.12. The expression pattern of LiMCT gene was found to be consistent with the accumulation sites and emission patterns of floral fragrance monoterpenes in transcriptome data (unpublished). Subcellular localization indicated that the LiMCT protein is located in chloroplasts, which is consistent with the location of MEP pathway genes functioning in plastids to produce isoprene precursors. Overexpression of LiMCT in Arabidopsis thaliana affected the expression levels of MEP and MVA pathway genes, suggesting that overexpression of the LiMCT in A. thaliana affected the metabolic flow of C5 precursors of two different terpene synthesis pathways. The expression of the monoterpene synthase AtTPS14 was elevated nearly fourfold in transgenic A. thaliana compared with the control, and the levels of carotenoids and chlorophylls, the end products of the MEP pathway, were significantly increased in the leaves at full bloom, indicating that LiMCT plays an important role in regulating monoterpene synthesis and in the synthesis of other isoprene-like precursors in transgenic A. thaliana flowers. However, the specific mechanism of LiMCT in promoting the accumulation of isoprene products of the MEP pathway and the biosynthesis of floral monoterpene volatile components needs further investigation.

Genome-Wide Characterization of Differentially Expressed Scent Genes in the MEP Control Network of the Flower of Lilium 'Sorbonne'.

Fragrance is an important feature of ornamental lilies. Components of volatile substances and important genes for monoterpene synthesis in the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway were examined in this study. Twenty volatile compounds (2 in the budding stage, 3 in the initial flowering stage, 7 in the semi-flowering stage, 17 in the full-flowering stage, and 5 in withering stage) were detected in the Oriental lily 'Sorbonne' using gas chromatography-mass spectrometry. The semi- and full-flowering stages were key periods for volatile substance production and enzyme function. Sequence assembly from samples collected during all flowering stages resulted in the detection of 274,849 genes and 129,017 transcripts. RNA sequencing and heatmapping led to the detection of genes in the MEP monoterpene metabolism pathway. Through gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis, we extracted key genes (LiDXS2, LiLIS, and LiMYS) and transcription factors (in the bHLH, MYB, HD-ZIP, and NAC families) associated with the MEP pathway. Tissue localization revealed that LiDXS2, LiLIS, and LiMYS were expressed in Lilium 'Sorbonne' petals in the full-flowering stage. Genes regulating the 1-deoxy-D-X-lignone-5-phosphate synthase family of rate-limiting enzymes, involved in the first step of monoterpene synthesis, showed high expression in the semi- and full-flowering stages. LiDXS2 was cloned and localized in chloroplast subcells. The relative expression of terpene-related genes in the MEP and mevalonic acid pathways of wild-type and LiLIS/LiMYS transgenic Arabidopsis thaliana, and changes in chemical composition, confirmed that LiLIS/LiMYS regulates the monoterpene synthesis pathway. The results of this study provide a theoretical basis for the synthesis of lily aromatic substances and the cultivation of new garden flower varieties.

Novel mechanism by which extracellular vesicles derived from Lactobacillus murinus alleviates deoxynivalenol-induced intestinal barrier disruption.

Deoxynivalenol (DON) is a common environmental pollutant that poses a serious health risk to humans worldwide. This study was aim to explore whether gut microbiota is involved in DON-induced intestinal toxicity as well as to reveal effect of probiotics derived from gut microbiota in protecting intestinal barrier and to elucidate mechanism. We found that DON caused disturbed gut microbiota, particularly Lactobacillus murinus (L. murinus) deficiency. DON enhanced M1 macrophage polarization and decreased tight junction protein expression. Microbiota transplantation experiments showed that transfer of DON-disrupted microbiota to healthy mice resulted in delivery of DON-induced intestinal toxicity. Besides, DON lost its damaging effect on macrophage and intestinal barrier in antibiotic-treated mice. Further intervention experiments revealed that L. murinus induce macrophage conversion from M1 to M2 phenotype through secreted extracellular vesicles (EVs) to alleviate DON-induced intestinal barrier disruption. Mechanistically, EVs activate TLR2 to promote M2 macrophage polarization and release IL-10, which in turn enhances intestinal barrier function. Upon successful translation of its efficacy into clinical practice, EVs created from L. murinus could be a novel possible treatment strategy for DON-induced gut disease.

Extracellular vesicles derived from Lactobacillus johnsonii promote gut barrier homeostasis by enhancing M2 macrophage polarization.

INTRODUCTION: Diarrheic disease is a common intestinal health problem worldwide, causing great suffering to humans and animals. Precise manipulation strategies based on probiotics to combat diarrheic diseases have not been fully developed. OBJECTIVES: The aim of this study was to investigate the molecular mechanisms by which probiotics manipulate macrophage against diarrheic disease. METHODS: Metagenome reveals gut microbiome profiles of healthy and diarrheic piglets. Fecal microbial transplantation (FMT) was employed to explore the causal relationship between gut microbes and diarrhea. The protective role of probiotics and their derived extracellular vesicles (EVs) was investigated in ETEC K88-infected mice. Macrophage depletion was performed to assess the role of macrophages in EVs against diarrhea. Execution of in vitro cell co-culture and transcriptome analyses elucidated the molecular mechanisms by which EVs modulate the macrophage and intestinal epithelial barrier. RESULTS: Escherichia coli was enriched in weaned diarrheic piglets, while Lactobacillus johnsonii (L. john) showed a negative correlation with Escherichia coli. The transmission of diarrheic illness symptoms was achieved by transferring fecal microbiota, but not metabolites, from diarrheic pigs to germ-free (GF) mice. L. john's intervention prevented the transmission of disease phenotypes from diarrheic piglets to GF mice. L. john also reduces the gut inflammation induced by ETEC K88. The EVs secreted by L. john demonstrated enhanced efficacy in mitigating the adverse impacts induced by ETEC K88 through the modulation of macrophage phenotype. In vitro experiments have revealed that EVs activate M2 macrophages in a manner that shuts down ERK, thereby inhibiting NLRP3 activation in intestinal epithelial cells. CONCLUSION: Our results reveal that intestinal microbiota drives the onset of diarrheic disease and that probiotic-derived EVs ameliorate diarrheic disease symptoms by modulating macrophage phenotypes. These findings can enhance the advancement of innovative therapeutic approaches for diarrheic conditions based on probiotic-derived EVs.

Physical Model of Inclusions Removal at Static Steel-Slag Interface.

Inclusions are one of the important factors affecting the cleanliness of molten steel. The current optimization of inclusion removal methods mainly focuses on promoting inclusions to float to the slag-steel interface so that the inclusions can be absorbed and removed by the refining slag. However, the research on the floating removal of inclusions cannot be carried out directly in the ladle, so methods such as mathematical models and physical models were developed. This article uses silicone oil to simulate the slag layer; polypropylene particles; and aluminum oxide particles to simulate inclusions to establish a water model experiment. By changing the viscosity of silicone oil and the diameter of particles, the factors affecting the movement of inclusions at the slag-steel interface were explored. Based on the water model, a mathematical model of the floating behavior of inclusions at the slag-steel interface was constructed, and parameters such as particle diameter and interfacial tension in the water model experiment were studied by the mathematical model for calculation. Both the mathematical model and the water model experimental results show that after the viscosity of silicone oil increases from 0.048 Pa·s to 0.096 Pa·s, the dimensionless displacement and terminal velocity of the particles decreases. When the diameter of the same particle increases, the dimensionless displacement and terminal velocity increases. The dimensionless displacement of polypropylene particles of the same diameter is larger than that of aluminum oxide particles, and the terminal velocity is smaller than that of aluminum oxide particles. This is attributed to the better overall three-phase wettability of polypropylene particle. When the liquid level increases, the dimensionless displacement and terminal velocity of particles under the same conditions show only slight differences (less than 10%).

Phase shift extraction algorithm based on Euclidean matrix norm.

In this Letter, the character of Euclidean matrix norm (EMN) of the intensity difference between phase-shifting interferograms, which changes in sinusoidal form with the phase shifts, is presented. Based on this character, an EMN phase shift extraction algorithm is proposed. Both the simulation calculation and experimental research show that the phase shifts with high precision can be determined with the proposed EMN algorithm easily. Importantly, the proposed EMN algorithm will supply a powerful tool for the rapid calibration of the phase shifts.

Massilia lurida sp. nov., isolated from soil.

A bacterial isolate, designated strain D5(T), was isolated from a soil sample collected from the Inner Mongolia Autonomous Region, China, and subjected to a taxonomic investigation using a polyphasic approach. Strain D5(T) was aerobic, Gram-stain-negative, rod-shaped and motile. Strain D5(T) fell within the evolutionary radius of the genus Massilia in the phylogenetic tree based on 16S rRNA gene sequences and was most closely related to Massilia plicata 76(T) with 97.3% 16S rRNA gene sequence similarity. The predominant quinone of strain D5(T) was Q-8. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The major fatty acids were summed feature 3 (comprising C16:1ω7c and/or iso-C15:0 2-OH) and C16:0. These chemotaxonomic data supported the affiliation of strain D5(T) to the genus Massilia. The genomic DNA G+C content was 65.9 mol%. Mean DNA-DNA relatedness values between strain D5(T) and the phylogenetically most closely related species of the genus Massilia, Massilia plicata KCTC 12344(T) and Massilia dura KCTC 12342(T), were 26 and 21%, respectively. Strain D5(T) could be differentiated from recognized species of the genus Massilia by several phenotypic characteristics. It is clear from the data presented that strain D5(T) represents a novel species of the genus Massilia, for which the name Massilia lurida sp. nov. is proposed. The type strain is D5(T) (=CGMCC 1.10822(T)=KCTC 23880(T)).

Three Strains of Lactobacillus Derived from Piglets Alleviated Intestinal Oxidative Stress Induced by Diquat through Extracellular Vesicles.

Previous studies found that Poria cocos polysaccharides (PCPs) significantly enhanced the antioxidant activity in piglet intestines while increasing the abundance of Lactobacillus. However, the relationship between Lactobacillus and antioxidant activity has yet to be verified, and the mode of action needs further investigation. Six Lactobacillus strains isolated from the intestines of neonatal piglets fed with PCPs were studied to investigate the relationship between Lactobacillus and intestinal oxidative stress. The results showed that three of them alleviated intestinal oxidative stress and protected the intestinal barrier. Subsequently, we extracted the extracellular vesicles (EVs) of these three Lactobacillus strains to verify their intestinal protection mode of action. We found that these EVs exerted an excellent antioxidant effect and intestinal barrier protection and could directly improve intestinal microbial composition. Our findings suggested that the EVs of the three Lactobacillus strains could enhance antioxidant activity by improving the physical intestinal barrier and remodeling gut microbiota. Unlike probiotics, which should be pre-colonized, EVs can act directly on the intestines. This study provides new ideas for the subsequent development of products to protect intestinal health.

Agrobacterium tumefaciens-Mediated Genetic Transformation of Narrowleaf Plantain.

Species in the genus Plantago have several unique traits that have led to them being adapted as model plants in various fields of study. However, the lack of a genetic manipulation system prevents in-depth investigation of gene function, limiting the versatility of this genus as a model. Here, a transformation protocol is presented for Plantago lanceolata, the most commonly studied Plantago species. Using Agrobacterium tumefaciens-mediated transformation, 3 week-old roots of aseptically grown P. lanceolata plants were infected with bacteria, incubated for 2-3 days, and then transferred to a shoot induction medium with appropriate antibiotic selection. Shoots typically emerged from the medium after 1 month, and roots developed 1-4 weeks after the shoots were transferred to the root induction medium. The plants were then acclimated to a soil environment and tested for the presence of a transgene using the ß-glucuronidase (GUS) reporter assay. The transformation efficiency of the current method is ~20%, with two transgenic plants emerging per 10 root tissues transformed. Establishing a transformation protocol for narrowleaf plantain will facilitate the adoption of this plant as a new model species in various areas.

Sodium arsenite exposure enhances H3K14 acetylation and impairs male spermatogenesis in rat testes.

Histone modifications play important roles in the epigenetic regulation of spermatogenesis via mediating gene transcription. Steroidogenic regulatory enzymes control testosterone biosynthesis, which are essential for spermatogenesis. Arsenic exposure inhibits the expression of steroidogenic genes by significantly increasing tri-methylation of H3K9 (H3K9me3) level in rat testis, finally diminishes testosterone release and lowers the rat sperm quality. Acetylation of H3K14 (H3K14ac) is associated with testosterone production and spermatogenesis. Co-occurrence of H3K9me3/H3K14ac has been identified previously by mass spectrometry in histone H3 isolated from different human cell types. H3K9me3/H3K14ac dually marked regions are in a poised inactive state to inhibit the gene expression. Whereas, whether inorganic arsenic exposure affects spermatogenesis and steroidogenic regulatory enzymes via mediating H3K14ac level has not been studied. Thereupon, the male Sprague-Dawley (SD) rats were exposed to (NaAsO2) for 6 weeks, then the sperm density and motility, testosterone level in serum, arsenic in rat testis were detected. mRNA expression of steroidogenic regulatory enzymes Star, Cyp11a1, Hsd3b and Hsd17b were determined by RT-PCR. H3K14ac level and the expression of histone acetylases of H3K14 (KAT2A and EP300), histone deacetylases of H3K14 (HDAC6 and HDAC3), the reader of H3K14ac (BAZ2A) were determined. The results suggested arsenic enhances H3K14ac in rat testis, which was associated with repression of steroidogenic regulatory genes expression, further reduced testosterone production, and impaired the spermatogenesis.

Mycobacterium litorale sp. nov., a rapidly growing mycobacterium from soil.

A gram-positive, acid-fast and rapidly growing rod, designated F4(T), was isolated from a soil sample of Haikou in China. The isolate shared 98.2 % 16S rRNA gene sequence similarity with Mycobacterium monacense B9-21-178(T), 96.2 % hsp65 sequence similarity with M. monacense FI-05352 and 79.6 % 16S-23S rRNA internal transcribed spacer sequence similarity with M. monacense B9-21-178(T). DNA-DNA relatedness between the isolate and M. monacense DSM 44395(T) was 43.5 %. The morphological analysis and physiological tests also showed that the isolate differed from any strain reported to date. The mycolic acid profile and the cellular fatty acid composition were also determined. On the basis of phenotypic and chemotaxonomic characteristics and phylogenetic data, it was concluded that strain F4(T) ( = CGMCC 4.5724(T) = JCM 17423(T)) merited classification as the type strain of a novel species, for which the name Mycobacterium litorale sp. nov. is proposed.

Massilia flava sp. nov., isolated from soil.

A Gram-stain-negative, rod-shaped bacterium, designated strain Y9(T), was isolated from a soil sample collected in Ningxia Province in China and was characterized to determine its taxonomic position. Strain Y9(T) contained Q-8 as the predominant ubiquinone. Major fatty acid components were summed feature 3 (C(16:1)ω7c and/or iso-C(15:0) 2-OH) and C(16:0). The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The G+C content of the genomic DNA of strain Y9(T) was 68.7 mol%. A phylogenetic tree based on 16S rRNA gene sequences showed that the strain fell within the evolutionary radiation encompassed by the genus Massilia. Levels of 16S rRNA gene sequence similarity between strain Y9(T) and the type strains of recognized Massilia species ranged from 95.2 to 98.2%, the highest values being with Massilia albidiflava 45(T) (98.2%) and Massilia lutea 101(T) (98.0%). However, levels of DNA-DNA relatedness between strain Y9(T) and M. albidiflava KCTC 12343(T) and M. lutea KCTC 12345(T) were 37 and 26%, respectively. Strain Y9(T) was clearly differentiated from its nearest phylogenetic relatives in the genus Massilia based on phenotypic, chemotaxonomic and phylogenetic properties. Therefore, strain Y9(T) is considered to represent a novel species of the genus Massilia, for which the name Massilia flava sp. nov. is proposed. The type strain is Y9(T) (=CGMCC 1.10685(T) =KCTC 23585(T)).

Heterologous Expression of LiSEP3 from Oriental Lilium Hybrid 'Sorbonne' Promotes the Flowering of Arabidopsis thaliana L.

The MADS-box gene family has multiple molecular and biological functions in plants. Here, the LiSEP3 gene of the MADS-box gene family of' 'Sorbonne' was obtained by homologous cloning using the petals of the flowering stage of Lilium Oriental Hybrid 'Sorbonne.' The ORF full-length sequence is 729 bp, encoding 242 amino acids. Bioinformatics analysis showed that the relative molecular weight of the LiSEP3 protein is 27.67 kD and the isoelectric point (pI) is 9.16. The prediction result of the gene positioning is transcription in its nucleus. Homologous alignment of amino acid sequences showed that the protein not only had typical MADS-box and K-box domains, but also contained two short and relatively conservative SEP motifs. The phylogenetic tree showed that the amino acid sequence encoded by the LiSEP3 gene had the closest relationship with SEP3 in monocotyledon plants such as Apostasia odorata. The results of real-time PCR showed that LiSEP3 gene was mainly expressed in petal. During flower development, the expression level of the LiSEP3 gene showed an overall trend of initially increasing and then decreasing. The flowering time of LiSEP3 transgenic Arabidopsis thaliana L. plants was earlier than that of wild-type Arabidopsis thaliana L. plants, compared with wild type, the number of rosette leaves is less. In the transgenic plants, the expression of flowering-associated AtSPL5 and AtGI genes was up-regulated, while the expression of AtSVP and AtFRI genes that inhibit flowering was down-regulated, which was consistent with the statistical results of the flowering time of LiSEP3 transgenic plants. Our results illustrate that the heterologous expression of SEP3 functional genes in the MADS-box family promoted the flowering period of transgenic plants of this hybrid. This research provides a theoretical basis for improving the flowering period of ornamental plants through plant genetic engineering technology and enhancing their economic and social values.

Overexpression of Nicotianamine Synthase (AtNAS1) Increases Iron Accumulation in the Tuber of Potato.

Iron (Fe) deficiency is a global health problem, especially in underdeveloped countries. Biofortification with genetic engineering methods has been used to improve Fe nutrition in a number of crops. Various steps, e.g., uptake, distribution, and storage, involved in Fe homeostasis have been manipulated to increase the Fe concentration in the edible portions of plants. Nicotianamine (NA) is an important metal ion chelator in plants. It promotes the mobility of Fe and decreases cellular Fe toxicity. Increasing the Fe content in crops by promoting NA synthesis could help decrease human diseases associated with Fe deficiency. In the present study, Arabidopsis thaliana nicotianamine synthase 1 (AtNAS1) was overexpressed in potato (Solanum tuberosum, St) under the control of the cauliflower mosaic virus 35S promoter. Transgenic plants had a significantly increased amount of Fe in tubers (52.7 µg/g dry weight, 2.4-fold the amount in wild-type tubers), while no differences in plant phenotype or yield were detected between transgenic and wild-type plants. The expression of genes involved in root mineral uptake and homeostasis, such as StYSL1, StIRT1, StFRO1, and StNAS, was also altered in the roots and leaves of the transgenic plants. Our results demonstrate that the manipulation of Fe chelation is a useful strategy for Fe nutrition improvement, and the increased Fe accumulation in tubers of transgenic potato plants is most likely caused by the increased movement of Fe from the leaf to the tuber.

Moxifloxacin is a safe and effective candidate agent for tuberculosis treatment: a meta-analysis of randomized controlled trials.

BACKGROUND: Moxifloxacin is a fourth-generation fluoroquinolone that has shown good antibacterial activity against both gram-positive cocci and gram-negative bacteria. The purpose of this study was to evaluate the safety and efficacy of moxifloxacin in the drug treatment regimen of patients with tuberculosis. METHODS: We conducted an electronic database search of the PubMed, Embase, the Cochrane Controlled Center Register of Controlled Trials (CENTRAL), Web of Science, Baidu Scholar, and Google Scholar for literature related to clinical randomized controlled trials (RCTs) of tuberculosis patients (from the date of inception of the database to September 25, 2020). The experimental group received moxifloxacin while the control group did not use moxifloxacin. After literature screening, data extraction, and literature quality evaluation, the included studies were meta-analyzed using RevMan software 5.1. RESULTS: In total, 13 RCTs involving 7,774 patients were included in this meta-analysis. The negative rate of sputum culture in the experimental group (which received moxifloxacin) was significantly higher than that of the control group after 2 months of treatment [relative risk (RR) =1.12, 95% confidence interval (CI): 1.06-1.18, P<0.0001]. Treatment-related complications in the experimental group were significantly greater than those in the control group (RR =1.34, 95% CI: 1.07-1.67, P=0.01). There was no significant difference in the incidence of serious complications between the two groups (RR =1.09, 95% CI: 0.90-1.33, P=0.38). In addition, there were no significant differences in mortality and recurrence rate between the two groups (RR =0.79, 95% CI: 0.51-1.21, P=0.28; RR =1.41, 95% CI: 0.61-3.25, P=0.42). CONCLUSIONS: This meta-analysis found that the addition of moxifloxacin to the treatment regimen of pulmonary tuberculosis patients could significantly increase the negative rate of sputum culture after treatment; however, it has no significant effect on the recurrence rate. Also, the addition of moxifloxacin was found to increase the incidence of complications, but did not increase the incidence of mortality or serious complications.