Hypo-osmolality activates Wnt/ß-catenin mediated by AQP1 in nucleus pulposus cells degeneration.

The goals of this study were to investigate whether Wnt/ß-catenin signaling plays a role in hypo-osmolality-related degeneration of nucleus pulposus (NP) cells, and if so, to define the mechanism underlying AQP1 in this effect. Human NP cells were cultured under hypo-osmotic (300/350/400 mOsm) and iso-osmotic (450 mOsm) conditions. The cell viability, AQP1, the expression of Wnt/ß-catenin signaling, collagen II/I, and MMP3/9 were evaluated. To determine the effects of the Wnt/ß-catenin signaling, we used the inhibitor and the activator of Wnt during the hypo-osmotic culture of NP cells. We also examined whether the silencing and overexpressing of the AQP1 gene would affect the Wnt/ß-catenin expression in NP cells. Hypo-osmolality caused NP cell degeneration and activated the Wnt/ß-catenin signaling but suppressed the AQP1 level. Inhibiting the Wnt/ß-catenin signaling alleviated the hypo-osmolality-induced NP cell degeneration. On the contrary, activating Wnt/ß-catenin aggravated the NP cell degeneration under hypo-osmotic conditions, which did not affect AQP1 expression. AQP1-overexpressed NP cells exhibited decreased Wnt/ß-catenin signaling and alleviated cell degeneration under the hypo-osmotic condition. Besides, AQP1 silencing accelerated NP cell degeneration and activated Wnt/ß-catenin expression compared with untreated control. Hypo-osmolality promotes NP cell degeneration via activating Wnt/ß-catenin signaling, which is suppressed by AQP1 expression. The upregulation of AQP1 suppressed the Wnt/ß-catenin signaling and alleviated the hypo-osmolality induced by the NP cell degeneration.

First Report of Leaf Spot on Cucumis melo L. Caused by Arcopilus aureus in China.

Cucumis melo L. is an important fruit with widespread consumption and commercial value. However, an undescribed disease affecting Hami melon (Cucumis melo L. var. Luhoutian) plants has consistently emerged in the Qihe region of Dezhou, Shandong Province of China since 2021. The disease can occur in both seedling and mature stages of Hami melon plants, and in severely diseased areas, the incidence rate was seen as 40 to 80%. During the seedling stage, the initial symptom is the appearance of water-soaked spots on the leaves. As the disease progresses, the leaves develop necrotic spots, and severely affected plants may exhibit stem rot and decay. In the mature stage, the disease primarily affects the leaves, causing necrotic spots and chlorosis. Under conditions of high humidity, black mold can be observed in the affected areas. Small pieces of symptomatic leaves from six different infected plants were collected and surface-sterilized with 5% NaClO for 3 min and 75% alcohol for 30 s for pathogen isolation (Wang et al., 2020). After rinsing with sterile water and blotted on sterile filter paper, the tissues were established on potato dextrose agar (PDA) media and incubated at 28℃ for 3-4 days. Pure isolates showed up at PDA were obtained through single-spore isolation. Colonies of all 16 isolates obtained by single-spore isolation had similar morphological characteristics on the PDA medium, the mycelium of the isolate appears dense and yellowish-brown on the PDA medium, and also secretes a brownish-red pigment on PDA. Under the opticalmicroscope, the perithecia from PDA media are subglobose spherical in shape, 80-100 µm in diameter, brownish by reflected light, wholly and densely hairy. Terminal hairs are very dense, greyish by reflected light, olive brown to reddish brown by transmitted light, thick-walled, arcuate, circinate, or spirally coiled at the apex. The ascospores within the perithecia are elliptical or droplet-shaped, initially colorless hyaline but later becoming subhyaline slightly gray, with dimensions of 7-9 µm × 4-5 µm. The morphological characteristics of the isolates were consistent with the description of Arcopilus aureus (Wang et.al. 2016). The internal transcribed spacer (ITS) region and ß-tubulin genes of three randomly selected isolates were PCR amplified and sequenced using primers ITS4/ITS5 and Bt2a/Bt2b. The sequences of ITS and ß-tubulin genes were submitted to NCBI with GenBank Accession No. OR539527 and OR640972, respectively. Based on morphological features and phylogenetic analysis, we concluded that the isolates belonged to A. aureus. Pathogenicity tests were conducted by placing agar plugs-containing fungal mycelia and agar blocks (control) on leaves of Hami melon seedlings (n=12) grown at 28°C with 60% humidity in a greenhouse, the assay was repeated three times. Symptoms appeared on the pathogen-inoculated leaves seven days after inoculation, whereas the control treatment remained symptomless. The pathogens were reisolated from diseased leaves and identified as A. aureus based on morphological, and molecular phylogenetic analysis, while Koch'sostulate was used to confirm its life mode. To the best of our knowledge, this is the first report of leaf spot caused by A. aureus on Cucumis melo L. in China.

[Design and Testing of a Pressure Measurement and Adjustment Device for Fracture Ends].

Objective: To design and test a device which is capable of accurately measuring and dynamically adjusting the axial pressure at the fracture end in real-time. Methods: Upon completion of the design, the pressure measurement and adjustment device was implemented in a canine tibial fracture external fixation model. A pressure sensor was mounted at the fracture end, and the displayed values of the pressure sensor were used as the standard for comparison. The relationship between the displayed values of the measurement and adjustment device and the pressure sensor under identical conditions was examined. Results: The device was utilized in external fixation models of tibial fractures in five beagles. A linear correlation was observed between the displayed values of the device and the pressure sensor at the fracture end. The measurement values from the device could be transformed into fracture end pressure through the application of coefficients, thereby facilitating accurate measurement and dynamic adjustment of the fracture end pressure. Conclusion: The pressure measurement and adjustment device at the fracture end is easy to operate, enabling precise measurement and dynamic regulation of the pressure at the fracture end. It is well-suited for animal experiments aimed at investigating the impact of axial compression on fracture healing, demonstrating promising potential for experimental applications.

Bacterial wilt affects the structure and assembly of microbial communities along the soil-root continuum.

BACKGROUND: Beneficial root-associated microbiomes play crucial roles in enhancing plant growth and suppressing pathogenic threats, and their application for defending against pathogens has garnered increasing attention. Nonetheless, the dynamics of microbiome assembly and defense mechanisms during pathogen invasion remain largely unknown. In this study, we aimed to investigate the diversity and assembly of microbial communities within four niches (bulk soils, rhizosphere, rhizoplane, and endosphere) under the influence of the bacterial plant pathogen Ralstonia solanacearum. RESULTS: Our results revealed that healthy tobacco plants exhibited more diverse community compositions and more robust co-occurrence networks in root-associated niches compared to diseased tobacco plants. Stochastic processes (dispersal limitation and drift), rather than determinism, dominated the assembly processes, with a higher impact of drift observed in diseased plants than in healthy ones. Furthermore, during the invasion of R. solanacearum, the abundance of Fusarium genera, a known potential pathogen of Fusarium wilt, significantly increased in diseased plants. Moreover, the response strategies of the microbiomes to pathogens in diseased and healthy plants diverged. Diseased microbiomes recruited beneficial microbial taxa, such as Streptomyces and Bacilli, to mount defenses against pathogens, with an increased presence of microbial taxa negatively correlated with the pathogen. Conversely, the potential defense strategies varied across niches in healthy plants, with significant enrichments of functional genes related to biofilm formation in the rhizoplane and antibiotic biosynthesis in the endosphere. CONCLUSION: Our study revealed the varied community composition and assembly mechanism of microbial communities between healthy and diseased tobacco plants along the soil-root continuum, providing new insights into niche-specific defense mechanisms against pathogen invasions. These findings may underscore the potential utilization of different functional prebiotics to enhance plants' ability to fend off pathogens.

Response of bacterial community metabolites to bacterial wilt caused by Ralstonia solanacearum: a multi-omics analysis.

The soil microbial community plays a critical role in promoting robust plant growth and serves as an effective defence mechanism against root pathogens. Current research has focused on unravelling the compositions and functions of diverse microbial taxa in plant rhizospheres invaded by Ralstonia solanacearum, however, the specific mechanisms by which key microbial groups with distinct functions exert their effects remain unclear. In this study, we employed a combination of amplicon sequencing and metabolomics analysis to investigate the principal metabolic mechanisms of key microbial taxa in plant rhizosphere soil. Compared to the healthy tobacco rhizosphere samples, the bacterial diversity and co-occurrence network of the diseased tobacco rhizosphere soil were significantly reduced. Notably, certain genera, including Gaiella, Rhodoplanes, and MND1 (Nitrosomonadaceae), were found to be significantly more abundant in the rhizosphere of healthy plants than in that of diseased plants. Eight environmental factors, including exchangeable magnesium, available phosphorus, and pH, were found to be crucial factors influencing the composition of the microbial community. Ralstonia displayed negative correlations with pH, exchangeable magnesium, and cation exchange flux, but showed a positive correlation with available iron. Furthermore, metabolomic analysis revealed that the metabolic pathways related to the synthesis of various antibacterial compounds were significantly enriched in the healthy group. The correlation analysis results indicate that the bacterial genera Polycyclovorans, Lysobacter, Pseudomonas, and Nitrosospira may participate in the synthesis of antibacterial compounds. Collectively, our findings contribute to a more in-depth understanding of disease resistance mechanisms within healthy microbial communities and provide a theoretical foundation for the development of targeted strategies using beneficial microorganisms to suppress disease occurrence.

[Short-term effect of black film covering on Oncomelania hupensis snail control in marshland and lake regions].

OBJECTIVE: To evaluate the short-term effect of the black film covering combined with niclosamide on Oncomelania hupensis snail control in marshland and lake regions. METHODS: A ditch with snails in Anxiang County was selected as the experimental sites. The ditch was divided into 3 groups: a film group, a niclosamide group, and a niclosamide and film group. The snails were surveyed before the test and 1, 3, 5, 7 days after the experiment. RESULTS: After the film covering, the highest temperature of the surface of soil was 63.7 degrees C on the first day, and the temperature higher than 40 degrees C lasted 4.34 hours. On the second day, the highest temperature was 52.3 degrees C, and the temperature higher than 40 degrees C lasted 4.96 hours. On the fourth day and fifth day, the temperature was lower than 30 degrees C, while the temperature was lower than 34 degrees C on the sixth day. On the seventh day of the experiment, the snail death rates of the film group, the niclosamide group, and the niclosamide and film group on the soil surface were 16.36%, 58.40% and 53.57%, respectively. On the seventh day, the snail death rates of the film group, and the niclosamide and film group under the soil were 20.00% and 40.00%, respectively, while no snails were found under the soil in the niclosamide group 3 days after the experiment. In mesh bags, the snail death rates of the film group, the niclosamide group, and the niclosamide and film group were 84.00%, 95.33% and 95.33%, respectively. CONCLUSION: The short-term effect of the black film covering on snail control is not obvious, and the black film covering does no promote the molluscicidal effect of niclosamide.

[Study on spatial-temporal variation of infected snail in bottomland areas after an integrated control strategy at village level in the marshland and lake regions based on geographic information system].

OBJECTIVE: To evaluate the effect of an integrated control strategy and to quantify the spatial-temporal variation of infected snails in the bottomland areas after the strategy was implemented. METHODS: Based on the geographic database of infected snail distribution at the village level during 2004 - 2010 in Anxiang county, Hunan province, spatial autocorrelation analysis and spatial scan statistics were applied to analyze the spatial-temporal characteristics on the distribution of infected snails. RESULTS: The number of embankments with infected snails in Anxiang county decreased from 23 in 2004 to 10 in 2010, while the rate of frame with infected snail in embankments decreased from 4.32‰ in 2004 to 0.12‰ in 2010. The spatial distribution of infected snails was nonrandom, only in 2004 and 2005 with Moran's I = 0.21 (P < 0.10) and Moran's I = 0.13 (P < 0.10) respectively. Data from the local spatial auto-correlation analysis showed that the number of villages with H-H types of auto-correlation model had been gradually decreasing. The results of SaTScan statistics appeared the same as from the local spatial auto-correlation analysis, showing that the number of areas with increased risk was decreasing. CONCLUSION: The comprehensive measures with emphasis on infectious source control seemed effective for schistosomiasis control program. The current distribution characteristics provided us with evidence that the resource assignment could be more reasonably implemented so as to control schistosomiasis in a more effective way.