Structural and biochemical insights into small RNA 3' end trimming by Arabidopsis SDN1.

A family of DEDDh 3'→5' exonucleases known as Small RNA Degrading Nucleases (SDNs) initiates the turnover of ARGONAUTE1 (AGO1)-bound microRNAs in Arabidopsis by trimming their 3' ends. Here, we report the crystal structure of Arabidopsis SDN1 (residues 2-300) in complex with a 9 nucleotide single-stranded RNA substrate, revealing that the DEDDh domain forms rigid interactions with the N-terminal domain and binds 4 nucleotides from the 3' end of the RNA via its catalytic pocket. Structural and biochemical results suggest that the SDN1 C-terminal domain adopts an RNA Recognition Motif (RRM) fold and is critical for substrate binding and enzymatic processivity of SDN1. In addition, SDN1 interacts with the AGO1 PAZ domain in an RNA-independent manner in vitro, enabling it to act on AGO1-bound microRNAs. These extensive structural and biochemical studies may shed light on a common 3' end trimming mechanism for 3'→5' exonucleases in the metabolism of small non-coding RNAs

Metabonomic profiling of clubroot-susceptible and clubroot-resistant radish and the assessment of disease-resistant metabolites

Plasmodiophora brassicae causes a serious threat to cruciferous plants including radish (Raphanus sativus L.). Knowledge on the pathogenic regularity and molecular mechanism of P. brassicae and radish is limited, especially on the metabolism level. In the present study, clubroot-susceptible and clubroot-resistant cultivars were inoculated with P. brassicae Race 4, root hairs initial infection of resting spores (107 CFU/mL) at 24 h post-inoculation and root galls symptom arising at cortex splitting stage were identified on both cultivars. Root samples of cortex splitting stage of two cultivars were collected and used for untargeted metabonomic analysis. We demonstrated changes in metabolite regulation and pathways during the cortex splitting stage of diseased roots between clubroot-susceptible and clubroot-resistant cultivars using untargeted metabonomic analysis. We identified a larger number of differentially regulated metabolites and heavier metabolite profile changes in the susceptible cultivar than in the resistant counterpart. The metabolites that were differentially regulated in both cultivars were mostly lipids and lipid-like molecules. Significantly regulated metabolites and pathways according to the P value and variable important in projection score were identified. Moreover, four compounds, including ethyl α-D-thioglucopyranoside, imipenem, ginsenoside Rg1, and 6-gingerol, were selected, and their anti-P. brassicae ability and effects on seedling growth were verified on the susceptible cultivar. Except for ethyl α-D-thioglucopyranoside, the remaining could inhibit clubroot development of varing degree. The use of 5 mg/L ginsenoside Rg1 + 5 mg/L 6-gingerol resulted in the lowest disease incidence and disease index among all treatments and enhanced seedling growth. The regulation of pathways or metabolites of carbapenem and ginsenoside was further explored. The results provide a preliminary understanding of the interaction between radish and P. brassicae at the metabolism level, as well as the development of measures for preventing clubroot

ANALYSIS ON 113 CASES OF ADVERSE REACTIONS CAUSED BY Β-LACTAM ANTIBIOTICS

The objectives of this study were to learn about the characteristics and rules of the occurrence of adverse reactions caused by lactam antibiotics and provide a reference for clinical drug use. Methods: A retrospective study was made to analyse the 113 case reports of adverse reactions caused by β-lactam antibiotics collected in our hospital between 2007 and 2009. Results: 113 cases of ADR involved 17 kinds of β-lactam antibiotics, headed by ceftriaxone sodium. The most common manifestation was skin and accessory damage; nervous system and gastrointestinal system damage were also easier to find, and the administration route was mainly intravenous infusion. Conclusion: The clinical application of β-lactam antibiotics should pay attention to adverse reaction monitoring and rational drug use to reduce the incidence of adverse reactions

Exogenous melatonin mediates radish (Raphanus sativus) and Alternaria brassicae interaction in a dose-dependent manner

Radish (Raphanus sativus L.) is an economically important vegetable worldwide, but its sustainable production and breeding are highly threatened by blight disease caused by Alternaria brassicae. Melatonin is an important growth regulator that can influence physiological activities in both plants and microbes and stimulate biotic stress resistance in plants. In this study, 0-1500 μM melatonin was exogenously applied to healthy radish seedlings, in vitro incubated A. brassicae, and diseased radish seedlings to determine the effects of melatonin on host, pathogen, and host-pathogen interaction. At sufficient concentrations (0-500 μM), melatonin enhanced growth and immunity of healthy radish seedlings by improving the function of organelles and promoting the biosynthesis of antioxidant enzymes, chitin, organic acid, and defense proteins. Interestingly, melatonin also improved colony growth, development, and virulence of A. brassicae. A strong dosage-dependent effect of melatonin was observed: 50-500 μM promoted host and pathogen vitality and resistance (500 μM was optimal) and 1500 μM inhibited these processes. Significantly less blight was observed on diseased seedlings treated with 500 μM melatonin, indicating that melatonin more strongly enhanced the growth and immunity of radish than it promoted the development and virulence of A. brassicae at this treatment concentration. These effects of MT were mediated by transcriptional changes of key genes as identified by RNA-seq, Dual RNA-seq, and qRT-PCR. The results from this work provide a theoretical basis for the application of melatonin to protect vegetable crops against pathogens

The long noncoding RNA LINC15957 regulates anthocyanin accumulation in radish

Radish (Raphanus sativus L.) is an important root vegetable crop belonging to the Brassicaceae family. Anthocyanin rich radish varieties are popular among consumers because of their bright color and high nutritional value. However, the underlying molecular mechanism responsible for skin and flesh induce anthocyanin biosynthesis in transient overexpression, gene silencing and transcriptome sequencing were used to verify its function in radish anthocyanin accumulation, radish remains unclear. Here, we identified a long noncoding RNA LINC15957, overexpression of LINC15957 was significantly increased anthocyanin accumulation in radish leaves, and the expression levels of structural genes related to anthocyanin biosynthesis were also significantly increased. Anthocyanin accumulation and expression levels of anthocyanin biosynthesis genes were significantly reduced in silenced LINC15957 flesh when compared with control. By the transcriptome sequencing of the overexpressed LINC15957 plants and the control, 5,772 differentially expressed genes were identified. A total of 3,849 differentially expressed transcription factors were identified, of which MYB, bHLH, WD40, bZIP, ERF, WRKY and MATE were detected and differentially expressed in the overexpressed LINC15957 plants. KEGG enrichment analysis revealed the genes were significant enriched in tyrosine, L-Phenylalanine, tryptophan, phenylpropanol, and flavonoid biosynthesis. RT-qPCR analysis showed that 8 differentially expressed genes (DEGs) were differentially expressed in LINC15957-overexpressed plants. These results suggested that LINC15957 involved in regulate anthocyanin accumulation and provide abundant data to investigate the genes regulate anthocyanin biosynthesis in radish

Dietary Corn Bran Altered the Diversity of Microbial Communities and Cytokine Production in Weaned Pigs

Corn bran (CB) has been used as an ingredient for pigs, but the underlying mechanisms that improve gut health is less clear. This study was conducted to investigate effects of dietary CB on growth performance, nutrient digestibility, plasma indices related to gut hormones and immunity, gut microbiota composition, and fermentation products in weaned pigs. A total of 60 weaned pigs were allocated to two dietary treatments, and piglets in each group received control (CON) diet or 5% CB diet for 28 days. Growth performance, nutrient digestibility, indices of gut hormones and immunity in plasma were evaluated. Microbiota composition in feces was determined using 16S rRNA amplicon sequencing, and fermentation products were measured by high-performance ion chromatography. The results showed that dietary CB did not affect growth performance, nutrient digestibility, gut hormones, or fermentation products in the trial (P > 0.05). There was an increased response to CB inclusion on interleukin-10 production (P < 0.05). On day 28, piglets fed dietary CB had a higher shannon index (P < 0.05). The population of the Firmicutes in CB treatment were decreased (P < 0.05), while the percentage of the Bacteroidetes were increased (P < 0.05). In particular, the populations of Eubacterium corprostanoligenes, Pevotella, and Fibrobacter related to polysaccharide fermentation of cereal bran were increased (P < 0.05). In conclusion, a post-weaning diet containing 5% CB increased intestinal microbial diversity, especially higher richness of fibrolytic bacteria, and promoted anti-inflammatory response to some extent in piglets, these changes should facilitate the adaptation of the digestive system of piglets in the subsequent growing phases

Widespread GLI expression but limited canonical hedgehog signaling restricted to the ductular reaction in human chronic liver disease

Canonical Hedgehog (Hh) signaling in vertebrate cells occurs following Smoothened activation/translocation into the primary cilia (Pc), followed by a GLI transcriptional response. Nonetheless, GLI activation can occur independently of the canonical Hh pathway. Using a murine model of liver injury, we previously identified the importance of canonical Hh signaling within the Pc+ liver progenitor cell (LPC) population and noted that SMO-independent, GLI-mediated signals were important in multiple Pc-ve GLI2+ intrahepatic populations. This study extends these observations to human liver tissue, and analyses the effect of GLI inhibition on LPC viability/gene expression. Human donor and cirrhotic liver tissue specimens were evaluated for SHH, GLI2 and Pc expression using immunofluorescence and qRT-PCR. Changes to viability and gene expression in LPCs in vitro were assessed following GLI inhibition. Identification of Pc (as a marker of canonical Hh signaling) in human cirrhosis was predominantly confined to the ductular reaction and LPCs. In contrast, GLI2 was expressed in multiple cell populations including Pc-ve endothelium, hepatocytes, and leukocytes. HSCs/myofibroblasts (gt;99%) expressed GLI2, with only 1.92% displaying Pc. In vitro GLI signals maintained proliferation/viability within LPCs and GLI inhibition affected the expression of genes related to stemness, hepatocyte/biliary differentiation and Hh/Wnt signaling. At least two mechanisms of GLI signaling (Pc/SMOdependent and Pc/SMO-independent) mediate chronic liver disease pathogenesis. This may have significant ramifications for the choice of Hh inhibitor (anti-SMO or anti-GLI) suitable for clinical trials. We also postulate GLI delivers a pro-survival signal to LPCs whilst maintaining stemness

Experimental investigation on flow field around a flapping plate with single degree of freedom

Undesirable flapping motion of discs can cause the failure of swing check valves in nuclear passive safety systems. Time-resolved particle image velocimetry (PIV) was employed to investigate the flow characteristics around a free-to-rotate plate and the motion response, with the Reynolds numbers, based on the hydraulic diameter of the channel, from 1.32 × 104 to 3.95 × 104. Appreciable flapping motion (±3.52°) appeared at the Reynolds number of 2.6 × 104 with the frequency of 5.08 Hz. In the low-Reynolds-number case, the plate showed negligible flapping. In the high-Reynolds-number case, the deflection angle increased with reduced flapping amplitude. The torque from the fluid determined the flapping amplitude. In the low-Reynolds-number case, Karman vortices were absent. With increasing Reynolds numbers, Karman vortices developed behind the plate with larger deflection angles. Strong interaction between the wake flow from the leading and trailing edge of the plate was observed. Based on power spectrum density (PSD) analysis, the vortex shedding frequency coincided with the flapping frequency, and the amplitude was positively correlated to the strength of the vortices. Proper orthogonal decomposition (POD) modes evince that, in the case of appreciable motion, coherent structures exhibited a larger spatial scale, enhancing the magnitude of the external torque on the plate

Recent Advances in Light-Induced Thermoelastic Spectroscopy for Gas Sensing: A Review

Light-induced thermoelastic spectroscopy (LITES) is a promising optical approach for gas sensing, which uses a quartz tuning fork (QTF) as a photothermal detector, instead of a commercial photodetector. Since the QTF has the advantages of low cost, small size, high resonance frequency, high-quality factor (Q-factor), and a wide spectral response range, and the LITES sensor has received extensive attention and obtained great development. This review paper summarizes and discusses the advances of the QTF-based, state-of-the-art LITES gas sensing technique in recent years and presents the development prospects of LITES sensor in the future

Past, Present and Future of Bacillus Calmette-Guérin Vaccine Use in China

The BCG vaccine is prepared from a weakened strain of Mycobacterium bovis (M. bovis), a bacterium closely related to Mycobacterium tuberculosis (MTB), which causes tuberculosis (TB). The vaccine was developed over 13 years, from 1908 to 1921, in the French Institut Pasteur by Léon Charles Albert Calmette and Jean-Marie Camille Guérin, who named the product Bacillus Calmette–Guérin (BCG). BCG, the only licensed vaccine currently available to prevent TB, is given to infants at high risk of TB shortly after birth to protect infants and young children from pulmonary, meningeal, and disseminated TB. The BCG vaccine, one of the safest and most widely used live attenuated vaccines in the world, recently celebrated its 100th anniversary (from 1921 to 2021); its record of use in preventing TB in China is also approaching 100 years. In 2022, a new century of BCG vaccine immunization will begin. In this article, we briefly review the history of BCG vaccine use in China, describe its current status, and offer a preliminary outlook on the future of the vaccine, to provide BCG researchers with a clearer understanding of its use in China