First report of Lasiodiplodia pseudotheobromae causing soft rot of plum in China.

Plum (Prunus salicina) is one of the most important fruit tree species worldwide (Valderrama-Soto et al. 2021). In June 2023, the postharvest soft rot symptoms were observed on plum fruits in several fruit markets of Guiyang city, Guizhou province, China. The disease incidence in these markets ranged from 20 to 25% with 70% disease severity. Plum fruits showed rotting, which was characterized by water soaked fruit tissue, softening and presence of whitish mycelia four days post inoculation. In severe conditions, whole fruits become rotted and were covered with white fungal mycelia. Small sections (5 × 3 mm) from 6 diseased plum fruits were surface sterilized by using 75% ethanol for 30 s followed by 0.1% mercuric chloride solution for 5 min, rinsed three times with ddH2O, and then transferred onto potato dextrose agar (PDA) and incubated at 25 ± 2°C for three days. Three pure cultures (GUCC23-0001 to GUCC23-0003) were obtained by transferring a single hyphal tip to new PDA plates. Colonies of these isolates were grayish-white initially, gradually turning to whitish brown with fluffy aerial mycelia and uneven edges and finally turned to a dark gray colony after five days of inoculation. The pseudoparaphyses were hyaline, cylindrical, aseptate, and rounded at apex. Conidia were ellipsoidal, hyaline, unicellular, and 24.2 to 28.6 × 12.3 to 15.5 µm in size (n = 30) (Fig. S1), which were similar to the morphology of Lasiodiplodia pseudotheobromae (Alves et al. 2008). Furthermore, fungal DNA was extracted from fresh mycelia of PDA after seven days by using fungus genomic DNA extraction kit (Biomiga, USA). Partial DNA sequences from four loci including internal transcribed spacer (ITS), translation elongation factor 1-alpha (tef1), beta-tubulin (tub2), and polymerase II second largest subunit (rpb2) were amplified with ITS1 and ITS4 (White et al. 1990), EF1-688F and EF1-1251R (Alves et al. 2008), Bt2a and Bt2b (Glass and Donaldson 1995), and RPB2-LasF and RPB2-LasR, respectively (Cruywagen et al. 2017). GenBank accession numbers are OR361680, OR361681, OR361682 for ITS, OR423394, OR423395, OR423396 for tef1, OR423397, OR423398, OR423399 for tub2, and OR423391, OR423392, OR423393 for rpb2, and gene sequencing showed 99.6 to 100% identity with ex-type strain of L. pseudotheobromae (CBS 116459). Phylogenetic analysis also placed our isolates in a highly supported clade with the reference isolate of L. pseudotheobromae (Fig. S2). Another experiment was designed to confirm the pathogenicity test for additional confirmation. Five mm mycelial plugs of L. pseudotheobromae from a three day old culture on PDA were placed on five surface-sterilized and non-wounded plum fruits for 12 hours and incubated at 25°C ± 2°C for four days. Sterilized fungus free PDA plugs were used as a negative control. Mycelial plugs were removed after 12 hours following which whole fruits were incubated in plastic boxes at 25°C ± 2°C. The experiment was repeated twice. The pathogenicity was evaluated under control conditions in laboratory (relative humidity, 70 ± 5% and temperature 25 ± 5˚C). Plum fruits showed rotting, which was characterized by water soaked fruit tissue, softening and presence of whitish mycelia four days post inoculation. These symptoms and signs were similar to the initially observed symptoms on plums in the markets. No disease symptoms were observed on the control fruits. The re-isolated fungus obtained from inoculated plum fruits was very similar to those isolated from diseased samples in morphology, fulfilling Koch's postulates. To the best of our knowledge, this is the first report of L. pseudotheobromae causing postharvest fruit rot of plum in China. In 2022, the total planting area of plum was 1946.5 thousand hectares, which produces approximately 6626300 tons of plum (Food and Agriculture Organization of the United Nations, 2022). Based on the disease incidence and severity reported in the current study, soft rot of plum may be responsible for nearly 35% of yield losses under severe. Therefore, our study laid a theoretical foundation for the prevention and control of this post-harvest disease of plum.

Unveiling the Potential of Bacillus safensis Y246 for Enhanced Suppression of Rhizoctonia solani.

Rhizoctonia solani is a significant pathogen affecting various crops, including tobacco. In this study, a bacterial strain, namely Y246, was isolated from the soil of healthy plants and exhibited high antifungal activity. Based on morphological identification and DNA sequencing, this bacterial strain was identified as Bacillus safensis. The aim of this investigation was to explore the antifungal potential of strain Y246, to test the antifungal stability of Y246 by adjusting different cultivation conditions, and to utilize gas chromatography-mass spectrometry (GC-MS) to predict the volatile compounds related to antifungal activity in Y246. In vitro assays demonstrated that strain Y246 exhibited a high fungal inhibition rate of 76.3%. The fermentation broth and suspension of strain Y246 inhibited the mycelial growth of R. solani by 66.59% and 63.75%, respectively. Interestingly, treatment with volatile compounds derived from the fermentation broth of strain Y246 resulted in abnormal mycelial growth of R. solani. Scanning electron microscopy analysis revealed bent and deformed mycelium structures with a rough surface. Furthermore, the stability of antifungal activity of the fermentation broth of strain Y246 was assessed. Changes in temperature, pH value, and UV irradiation time had minimal impact on the antifungal activity, indicating the stability of the antifungal activity of strain Y246. A GC-MS analysis of the volatile organic compounds (VOCs) produced by strain Y246 identified a total of 34 compounds with inhibitory effects against different fungi. Notably, the strain demonstrated broad-spectrum activity, exhibiting varying degrees of inhibition against seven pathogens (Alternaria alternata, Phomopsis. sp., Gloeosporium musarum, Dwiroopa punicae, Colletotrichum karstii, Botryosphaeria auasmontanum, and Botrytis cinerea). In our extensive experiments, strain Y246 not only exhibited strong inhibition against R. solani but also demonstrated remarkable inhibitory effects on A. alternata-induced tobacco brown spot and kiwifruit black spot, with impressive inhibition rates of 62.96% and 46.23%, respectively. Overall, these findings highlight the significant antifungal activity of B. safensis Y246 against R. solani. In addition, Y246 has an excellent antifungal stability, with an inhibition rate > 30% under different treatments (temperature, pH, UV). The results showed that the VOCs of strain Y246 had a strong inhibitory effect on the colony growth of R. solani, and the volatile substances produced by strain Y246 had an inhibitory effect on R. solani at rate of 70.19%. Based on these results, we can conclude that Y246 inhibits the normal growth of R. solani. These findings can provide valuable insights for developing sustainable agricultural strategies.

Biogenic Solution Map Based on the Definition of the Metabolic Correlation Distance between 4-Dimensional Fingerprints.

Accurate discrimination and classification of unknown species are the basis to predict its characteristics or applications to make correct decisions. However, for biogenic solutions that are ubiquitous in nature and our daily lives, direct determination of their similarities and disparities by their molecular compositions remains a scientific challenge. Here, we explore a standard and visualizable ontology, termed "biogenic solution map", that organizes multifarious classes of biogenic solutions into a map of hierarchical structures. To build the map, a novel 4-dimensional (4D) fingerprinting method based on data-independent acquisition data sets of untargeted metabolomics is developed, enabling accurate characterization of complex biogenic solutions. A generic parameter of metabolic correlation distance, calculated based on averaged similarities between 4D fingerprints of sample groups, is able to define "species", "genus", and "family" of each solution in the map. With the help of the "biogenic solution map", species of unknown biogenic solutions can be explicitly defined. Simultaneously, intrinsic correlations and subtle variations among biogenic solutions in the map are accurately illustrated. Moreover, it is worth mentioning that samples of the same analyte but prepared by alternative protocols may have significantly different metabolic compositions and could be classified into different "genera".

Luteolin ameliorates palmitate-induced lipotoxicity in hepatocytes by mediating endoplasmic reticulum stress and autophagy.

Abnormal accumulation of lipids in liver leads to uncontrolled endoplasmic reticulum (ER) stress and autophagy. Luteolin is known to have antioxidant, anti-inflammatory, and anti-cancer properties, but whether it protects against lipotoxicity in liver remains unclear. In this study, we challenged AML12 liver cells and mouse primary hepatocytes with palmitic acid (PA) with or without luteolin pretreatment. In the presence of PA, reactive oxygen species (ROS) production was increased at 3 h, followed by enhancement of expression of p-PERK, ATF4, p-eIF2α, CHOP, and TXNIP (ER stress markers) and p-p62 and LC3II/LC3I ratio (autophagy markers), in both primary hepatocytes and AML12 cells. When PA treatment was extended up to 24 h, apoptosis was induced as evidenced by an increase in caspase-3 activation. RFP-GFP-LC3B transfection further revealed that the fusion of autophagosomes with lysosomes was damaged by PA. With luteolin treatment, the expression of antioxidant enzymes, i.e., heme oxygenase-1 and glutathione peroxidase, was upregulated, and PA-induced ROS production, ER stress, and cell death were dose-dependently ameliorated. Luteolin could also reverse the damage caused to autophagic flux. These results indicate that luteolin protects hepatocytes against PA assault by enhancing antioxidant defense, which can attenuate ER stress and autophagy as well as promote autophagic flux.

Amelioration of 5-fluorouracil-induced intestinal mucositis by Streptococcus thermophilus ST4 in a mouse model.

Intestinal mucositis is a commonly encountered toxic side effect in patients undergoing 5-fluorouracil (5-FU)-based chemotherapy. Numerous studies have shown that probiotics enable improving chemotherapy-induced intestinal mucositis, but the beneficial effects of probiotics differ depending on the strain. Therefore, in the present studies we suggest that S. thermophilus ST4 separated from raw milk may assess mucoprotective activity in 5-FU-induced intestinal mucositis. In our causal-comparative study design, fifteen mice were randomized assigned into three groups (n = 5/each group): control group, 5-FU group and 5-FU+S. thermophilus ST4 group. The control group was orally administrated saline only, and the 5-FU group was followed by intraperitoneal injection of 5-FU for 3 days after 10-day saline administration, and the 5-FU+S. thermophilus ST4 group was intragastrically subjected for S. thermophilus ST4 once per day during the whole experiment, starting from the first day of the experiment, followed by 5-FU intraperitoneal injection for 3 days after 10-day S. thermophilus ST4 pretreatment. Diarrhea score, pro-inflammatory cytokines serum levels, intestinal histopathology and short chain fatty acid were assessed. Here, we demonstrated the beneficial effects of S. thermophilus ST4 derived from raw milk against 5-FU-induced intestinal mucositis, including body weight reduction, appetite loss and diarrhea. Intrinsically, S. thermophilus ST4 effectively maintained epithelium structure in small intestines and colons as well as reduced the intestinal inflammation. Besides, S. thermophilus ST4 significantly increased the expression of acetic acid, reinforcing the muco-protective effects. In conclusion, our results demonstrate that S. thermophilus ST4 supplementation ameliorates 5-FU-induced intestinal mucositis. This suggests probiotic may serve as an alternative therapeutic strategy for the prevention or management of 5-FU-induced mucositis in the future.

Defective minor spliceosomes induce SMA-associated phenotypes through sensitive intron-containing neural genes in Drosophila.

The minor spliceosome is evolutionarily conserved in higher eukaryotes, but its biological significance remains poorly understood. Here, by precise CRISPR/Cas9-mediated disruption of the U12 and U6atac snRNAs, we report that a defective minor spliceosome is responsible for spinal muscular atrophy (SMA) associated phenotypes in Drosophila. Using a newly developed bioinformatic approach, we identified a large set of minor spliceosome-sensitive splicing events and demonstrate that three sensitive intron-containing neural genes, Pcyt2, Zmynd10, and Fas3, directly contribute to disease development as evidenced by the ability of their cDNAs to rescue the SMA-associated phenotypes in muscle development, neuromuscular junctions, and locomotion. Interestingly, many splice sites in sensitive introns are recognizable by both minor and major spliceosomes, suggesting a new mechanism of splicing regulation through competition between minor and major spliceosomes. These findings reveal a vital contribution of the minor spliceosome to SMA and to regulated splicing in animals.

Numerous long single-stranded DNAs produced by dual amplification reactions for electrochemical detection of exosomal microRNAs.

Exosomal microRNAs (miRNAs) have been explored as an extremely promising biomarker of liquid biopsy for the diagnosis, treatment and prognosis of diseases such as cancer, in which sensitive and selective detection is significant. Herein, we describe the construction and testing of an electrochemical biosensor for the sensitive detection of exosomal miRNAs. It is based on synthetizing numerous long single-stranded DNAs (ssDNAs), which are produced by dual amplification reactions of target-triggered cyclic strand displacement reaction (TCSDR) and primer exchange DNA amplification reaction (PEDAR). In the first signal amplification step, target miRNAs are captured by the hairpin DNA strands (capture probes, Cp) that are immobilized on electrode. After strand unfolding with target capture, primer probes (Pp) enable to hybridize with Cp. And then target miRNAs were displaced for starting the TCSDR process that enable the introduction of numerous primers in Pp. In the second signal amplification step, the primers associated with PEDAR produce copious amounts of elongated ssDNAs. These ssDNAs absorb abundant quantities of methylene blue (MB) that enables the highly sensitive and label-free detection of exosomal miRNAs. This dual amplification process is characterized by a low limit of detection (LOD) of 3.04 aM. In addition, the electrochemical biosensor exhibits good selectivity for miR-21 detection, and shows benefits of simple operation, low cost, portability. Overall, the electrochemical biosensor provides a promising platform for the early diagnosis and screening of tumor biomarkers and the development of devices for point-of-care testing (POCT).

Rubiparvus bistigma, a new genus and species of Empoascini (Hemiptera, Cicadellidae, Typhlocybinae), with a checklist of the Alebroides group in Chinese fauna.

A new leafhopper genus of the tribe Empoascini, Rubiparvus gen. nov., is described based on Rubiparvus bistigma sp. nov. from Yunnan (southwest China). Habitus photos and illustrations of male genitalia of this new species are given. Differences between the new genus and closely related genera are discussed. A checklist of the Alebroides generic group in the Chinese fauna is provided.

[Clinical analysis of 91 cases of imported falciparum malaria from Africa].

OBJECTIVE: To explore the clinical characteristics, early diagnosis, and treatment of patients with imported falciparum malaria from Africa. METHODS: The clinical data of 91 imported falciparum malaria cases were analyzed by retrospective study. RESULTS: All the 91 cases had the history of mosquito bites. The clinical manifestation of these cases varied, including fever, headache, chill, diarrhea, anemia, thrombocytopenia, proteinuria, damage of liver function, abdominal ultrasonographic presentations (enlarged liver and spleen). All the patients were successfully treated with the combination therapy of artemether and primaquine. CONCLUSION: The key procedures for treating imported falciparum malaria are earlier diagnosis and effective therapy. The combination therapy with artemether and primaquine shows a high efficacy and low side effect and low relapsed rate.

[Soil respiration in subtropical forests and model simulation of its relationships with soil temperature and moisture content].

By using Li-6400-09 system, an in situ measurement on the soil respiration, soil temperature, and soil moisture content in three main forest types (evergreen broadleaved forest, Cunninghamia lanceolata plantation, and Phyllostachys pubescens forest) in subtropical area of China was conducted, with the relationships between soil respiration and soil temperature and moisture content simulated by several models. The C flux of soil respiration in P. pubescens forest, evergreen broadleaved forest, and C. lanceolata plantation was 12.84, 11.70, and 7.12 t C x hm(-2) x a(-1), respectively, and the soil respiration in these three forest types had similar diurnal and seasonal variations, with the maximum value at 11:00-12:00 and the minimum value at 1:00-3:00, and the highest value in August and September while the lowest value in December and January. Van't Hoff equation and Lloyd & Taylor function had less difference in describing the relationships between soil respiration and soil temperature, while the soil respiration rate predicted by Lloyd & Taylor function was smaller than the observed value. Quadratic model and power function model could well simulate the relationship between soil respiration and soil moisture content. Soil moisture content positively or negatively affected soil respiration, but the effects only reached significant level in C. lanceolata plantation. Comparing with single-factor equation, two-factor equation (soil temperature and moisture content) could better describe the responses of soil respiration to changed soil temperature and moisture content. Multivariate analysis of covariance showed that after eliminating the effects of soil temperature and moisture content, forest type had significant effects on soil respiration (R2 = 0.541). Other factors, such as air temperature, air relative humidity, and photosynthetic radiation also affected soil respiration, and the effects of air temperature reached significant level.

Fangchinoline inhibits glutamate release from rat cerebral cortex nerve terminals (synaptosomes).

Fangchinoline, an active component of radix stephaniae tetrandrinea, has been shown to possess neuroprotective properties. It has been reported that excessive glutamate release has been proposed to be involved in the pathogenesis of several neurological diseases. The primary purpose of the present study was to investigate the effect of fangchinoline on glutamate release in rat cerebral cortex nerve terminals and to explore the possible mechanism. Fangchinoline inhibited the release of glutamate evoked by 4-aminopyridine (4-AP) in a concentration-dependent manner, and this phenomenon resulted from a reduction of vesicular exocytosis but not from an inhibition of Ca(2+)-independent efflux via glutamate transporter. Fangchinoline did not alter the resting synaptosomal membrane potential or 4-AP-mediated depolarization, but significantly reduced depolarization-induced increase in [Ca(2+)](C). Fangchinoline-mediated inhibition of glutamate release was significantly prevented by the N- and P/Q-type Ca(2+) channel blocker omega-conotoxin MVIIC, and by the PKC inhibitors, GF109203X and Ro318220. In addition, the glutamate release mediated by direct Ca(2+) entry with Ca(2+) ionophore (ionomycin) was unaffected by fangchinoline, which suggests that the inhibitory effect of fangchinoline is not due to directly interfering with the release process at some point subsequent to Ca(2+) influx. These results suggest that fangchinoline inhibits glutamate release from the rat cortical synaptosomes through the suppression of voltage-dependent Ca(2+) channel activity and subsequent reduces Ca(2+) entry into nerve terminals, rather than any upstream effect on nerve terminal excitability. This inhibition appears to involve the suppression of PKC signal transduction pathway. This finding may explain the neuroprotective effects of fangchinoline against neurotoxicity.

[Study on thermogravimetric characteristics of simulated medical waste by using TG-DTA-FTIR analysis].

Thermogravimetric behavior of simulated medical waste was studied in the atmosphere of nitrogen and air at different heating rates by thermogravimetric analysis and differential thermal analysis (TG-DTA), and the influence on the thermogravimetric behavior of simulated medical waste by the atmosphere and heating rate was also discussed. The results indicate that the initial separating temperature of the volatile organic matter ( T(s)) moves toward higher temperature and the peak value of the weight loss velocity (DTGmax) has a remarkable increase with the increasing heating rate when simulated medical waste is heated at these two atmospheres. There are two weight loss peaks when the experimental material is pyrolyzed at the atmosphere of nitrogen and the corresponding weight loss rate is about 31% and 59% . There are three weight loss peaks at the atmosphere of air and the corresponding anterior two weight loss rate is about 44% - 59% and 31% - 46%. At the same time, qualitative analysis of evolved gases under different conditions was analyzed by the coupled simultaneous fourier transform infrared spectroscopy (FTIR). In addition, quantitative analysis of CO, CO2, H2O and CH4 was conducted. Experimental results show that the main thermogravimetric stages of simulated medical waste in the atmosphere of nitrogen and air have the similar products, and the typical absorbed peaks of CO2, CO, alkanes, aldehydes, carbonyl acids, alcohols and alkenes were detected. And result shows that the heating rate and atmosphere have an effect on the yield of CO, CO2, H2O and CH4. In the main stages of the process, the curves of water content changing along with the heating time show downward peaks, which count for the participation of water during the reaction.