Seasonal Succession of Fungal Communities in Native Truffle (Tuber indicum) Ecosystems.

Truffles are a rare underground fungus and one of the most expensive, and sought-after kitchen ingredients in the world. Microbial ecology plays an important role in the annual growth cycle of truffles, but fungal communities in native truffle ecosystems are still largely unknown, especially for Tuber indicum from China. In this study, the spatial and temporal dynamics of soil physicochemical properties and fungal communities were described associated with four T. indicum-producing plots (TPPs) and one non-truffle-producing plot in four successive growing seasons. A total of 160 biological samples were collected, 80 of which were used for the determination of 10 soil physicochemical indices and 80 for Illumina-based analysis of the fungal microbiome. Soil physicochemical properties and fungal communities exhibited considerable seasonal variation. Ascomycetes, Basidiomycetes, and Mucormycoides dominated. The core microbiome work on the microecological changes in TPPs, and the identified core members contribute to the seasonal succession of communities. The genus Tuber occupies a central position in healthy TPPs. There was a strong correlation between soil physicochemical properties and fungal communities. The genus Tuber showed a positive correlation with Ca, Mg, and total nitrogen, but a negative correlation with total phosphorus and available potassium. This study describes the complex ecological dynamics of soil physicochemical indices and fungal communities occurring during the annual cycle of Tuber indicum, and highlights the succession of core communities in truffle plots, which contribute to better protection of native truffle ecosystems and control of mycorrhizal fungal contamination in artificial truffle plantations in China. IMPORTANCE The spatial and temporal dynamics of soil physicochemical properties and fungal communities associated with four Tuber indicum-producing plots and one non truffle producing plot in four different growing seasons are described. Soil physicochemical properties and fungal communities exhibited considerable seasonal variation. This study examines the complex ecological dynamics of soil physicochemical indices and fungal communities occurring during the annual cycle of Tuber indicum and highlights the succession of core communities in truffle plots, which contributes to better protection of native truffle ecosystems and control of mycorrhizal fungal contamination in artificial truffle plantations in China.

Ultra-performance liquid chromatography-tandem mass spectrometry revealed the significantly different metabolic profiles of Auricularia cornea growing on weakly acidic and weakly alkaline substrates.

Auricularia cornea is a widely prized basidiomycetous mushroom with culinary and medicine value, which is cultivated artificially on a large scale in China. However, little attention has been paid to the differences in metabolic profiles under different pH growth conditions. In the present study, weakly acidic and weakly alkaline artificial substrates were developed and used for the cultivation of A. cornea, and the metabolic profiles of its fruiting bodies were determined using ultra performance liquid chromatography-tandem mass spectrometry. The results show that the weakly alkaline substrate environment promoted mycelial growth, increased body surface area, and improved yield and transformation efficiency, but attenuated metabolite accumulation by A. cornea. A total of 412 different metabolites were identified in negative and positive ion mode, of which 99 had significantly different amounts and covered 51 metabolic pathways. Principal component analysis and orthogonal patrial least squares discriminant analysis showed clear separation between two treatments, indicating different metabolic profiles. The different metabolites mainly included seven chemical categories, including amino acids and derivatives, nucleotides and derivatives, phenolic acids, organic acids, lipids, flavonoids and alkaloids. This study revealed the biological significance of these metabolites, which could be useful for further unexplored compounds and possible biological functions of A. cornea.

First Report of Brown Rot Disease on Cultivated Auricularia cornea Caused by Trichoderma pleuroticola in Sichuan Province, China.

Auricularia cornea Ehrenb. is a well-known, rare, and valuable edible mushroom, with considerable culinary and medicinal value. It is distributed worldwide and especially common in Asia (Khatua et al. 2022). In China, more than 1.89 million tons of A. cornea are artificially cultivated annually, particularly in Sichuan Province, which produces 0.97 million tons, accounting for about 51% of the nation's total (Ye et al. 2022). However, farmers in Shifang, a county-level city in Sichuan Province, who practiced traditional greenhouse cultivation on a large scale, reported that brown rot disease affected up to 20% of their A. cornea crop and caused severe failures each year between 2016 and 2022, resulting in devastating economic losses. Worse, when the diseased fruit bodies were not removed promptly by farmers, the disease could spread to 100% of crop in a given greenhouse in 10 to 15 days. The symptoms mainly occur in fresh unfolded fruiting bodies. Lesions were brown, putrefied, and foul smelling, and eventually the fruiting bodies wilted. Naturally air-dried fruiting bodies were dark brown with yellow deposits at the edges (Fig. 1A). In this backdrop, we collected more than 60 diseased samples from Shifang (104°1'15''E, 31°12' 30'' N) and isolated pathogens from May 2021 through May 2022. On the clean bench, the surfaces of the lesions were disinfected with cotton balls soaked in 75% ethanol and rinsed three times with sterile water, and then the internal tissue block was picked with a sterile scalpel and cultured in potato dextrose agar (PDA) medium at 25°C. The pathogen was repeatedly isolated and purified, and we conducted pathogenicity tests. Colonies of the pathogen on PDA medium were white and cottony, with a mycelial growth rate of 13.92 ± 1.24 mm/day at 25°C. Then the spores began to turn yellow-green and soon turquoise, converging into a wide concentric wheel. The spores were elliptical with dimensions of 4.0 to 8.0 µm × 3.0 to 4.5 µm (N=50). Pathogenicity tests were conducted in an incubator. Ninety pure white, healthy A. cornea fruiting bodies were randomly picked and placed in groups of 10. On the clean bench, the bodies surface were sterilized with 75% alcohol cotton balls, then washed three times with sterile water, dried with sterile absorbent paper, and placed in sterile petri dishes for subsequent tests. The first control group (CK1) did not receive additional treatment other than the surface disinfection mentioned above. These bodies were immediately sealed with parafilm. The second control group (CK2) was not inoculated with the pathogen, and a blank sterile PDA plug (diameter: 0.8 cm) was placed on the surface of the fruiting body and sealed with parafilm. All seven treatment groups (TS) were inoculated with single-pathogen mycelium plugs (diameter: 0.8 cm) in the center surface of each fruiting body. They were then sealed with parafilm. All total of nine experimental treatment groups were cultured in a light incubator at 25°C. The test results showed that the pathogen could visibly infect the A. cornea within 24 hours. At 48 hours after inoculation, the lesions were round and brown, radiating outward along the inoculum, consistent with the symptoms of the original sample. As culture time continued, the extent of each lesion gradually expanded (Fig. 1B). After 120 hours, the fruiting bodies showed rot, stench, and loss of other traits relevant to commercial value. In contrast, the CK1 and CK2 groups had no lesions. Furthermore, the fungal cultures with the same phenotypic characteristics could be continuously isolated from the lesions of TS, and the pathogenic factors were verified by Koch's postulates. Similarly, uninoculated and inoculated tests were conducted in a greenhouse, and the results were consistent with those of incubator testing. The pathogen was designated MMEBYJ202206. The DNA of the pathogen was extracted using CTAB, and the rDNA internal transcribed spacer (ITS) of the isolates was amplified using ITS1/ITS4 primers. PCR was conducted in a 25 µL reaction mixture, and a 596 bp sequence was obtained by sequencing. The ITS sequence has been submitted in GenBank with accession number ON974844.1. BLAST database in NCBI was used to compare ITS sequences and phylogenetic tree was constructed based on the neighbor-joining algorithm from MEGA (Fig. 2). The results indicate that the MMEBYJ202206 was located on a common clade of the phylogenetic tree with KX343129.1, KX343130.1, KX343131.1, and MF871554.1 of T. pleuroticola, and it showed 99% support in bootstrap (500 replicates), but it was not in the same clade as other Trichoderma spp., suggesting that the pathogen was T. pleuroticola. To our knowledge, this is the first report to show that T. pleuroticola can cause brown rot disease on artificially cultivated A. cornea. However, a previous study showed T. pleuroticola to be the causal agent of considerable decline in the yield of Pleurotus ostreatus and P. florida (Siwulski et al. 2011; Blaszczyk et al. 2013) and capable of infecting A. heimuer raised on artificial bed-log (a rod used for growing mycelium of edible mushrooms) (Liu et al. 2019). Brown rot disease is important in China because it has caused considerable damage to yield in artificially cultivated A. cornea, a decline in the external and internal qualities of the product, and a reduction in the enthusiasm of farmers for this crop. Consequently, this study provides a foundation for further research and prevention of this pathogen in China.

First Report of Vacuiphoma oculihominis Causing Fruit Rot on Navel Orange (Citrus sinensis) in China.

Newhall navel orange [Citrus sinensis (L.) Osbeck] is an economically important agricultural product in China. In February 2022, a rare lesion symptom was observed on Newhall navel oranges that were harvested from an orchard Ganzhou city, Jiangxi province, China (25.53° N, 114.79° E) and stored for 90 days (18±2℃, 80 to 90% RH) at the Jiangxi Key Laboratory for Postharvest Technology and Non-destructive Testing of Fruits and Vegetables (28.68° N, 115.85° E). Approximately 2% (15/750) of the oranges exhibited symptoms, with normal appearance but ink-black flesh and juice, yellowish lesions on edges of the symptoms, and no unusual odor. To isolate the pathogen, three 5 × 5 mm pieces of symptomatic tissue from a diseased orange were disinfected in 75% ethanol for 30 s, rinsed three times with sterile water, and inoculated on potato dextrose agar (PDA) at 25±1℃ and a 12:12 h photoperiod for 7 days. A pure isolate named ND-hsp was obtained. The colony was light yellow center with pale edge on the top and brown on the bottom. Conidia and pycnidia were observed on PDA medium after 2 months. Conidia were long oval, no septa, 2.9 × 3.4 µm (n = 50), and pycnidia were solitary, 39.4 × 43.9 µm (n = 20), with one or no orifice, brown to dark brown. The morphological characteristics of ND-hsp strain on PDA, oatmeal agar and malt extract agar were similar to those of the Didymellaceae (Aveskamp et al. 2010). Ulteriorly, the genomic DNA of the ND-hsp isolate was extracted from its mycelia using a fungal genomic DNA extraction kit (Solarbio, Beijing, China) for subsequent phylogenetic analyses. Four primer sets, LR0R (Rehner and Samuels 1994) /LR7 (Vilgalys and Hester 1990), V9G (Hoog and Gerrits 1998) /ITS4 (White et al. 1990), Btub2Fd/Btub4Rd (Woudenberg et al. 2009) and RPB2-5F2 (Sung et al. 2007)/RPB2-7cR (Liu et al. 1999) were used to amplify the corresponding DNA fragments of large subunit ribosomal RNA (LSU), internal transcribed spacer region (ITS), beta-tubulin gene (TUB2) and RNA polymerase Ⅱ second largest subunit (RPB2), respectively. The obtained sequences were assigned GenBank accession numbers and showed 99 to 100% identity with their counterparts of Vacuiphoma oculihominis UTHSC DI16-308. A phylogenetic tree was constructed in MEGA 7.0 using the concatenated sequences, placing the isolate within the V. oculihominis clade by 100% bootstrap support. Pathogenicity experiments were performed in triplicate. Ten Newhall navel oranges were surface sterilized with 75% ethanol and inoculated with 15µL of a spore suspension (2×106 spores/ml) into a 3 mm-diameter wound on the equator. The control group received sterile water instead of the spore suspension. Treated and control oranges were incubated at 25±1 ℃ and about 90% relative humidity for 20 days. All oranges were cut longitudinally or transversely through the inoculated wound and examined internally. The oranges inoculated with ND-hsp exhibited ink-black flesh and juice symptoms consistent with the initial oranges. The control oranges remained asymptomatic. Under the Koch's rule, V. oculihominis was reisolated from diseased oranges and kept in Collaborative Innovation Center of Postharvest Key Technology and Quality Safety of Fruits and Vegetables in Jiangxi Province. GenBank database analysis confirms that V. oculihominis has been found in human eye secretions and decayed trees. This is the first report of V. oculihominis as a pathogen on navel oranges in China. Our findings contribute to understanding of citrus fruit pathogens.

First Report of Fruit Rot Caused by Fusarium graminearum on Ponkan (Citrus reticulata Blanco cv. Ponkan) in China.

Ponkan (Citrus reticulata Blanco cv. Ponkan) is a Chinese citrus species with tasty fruit. In November 2021, an unknown postharvest disease of Ponkan fruit caused nearly 15% losses of 2000 fruits in Nanchang, Jiangxi Province (28.68° N, 115.85° E). The initial fruit's surface necrosis was brown (Xu et al. 2022) (Figure 1A). Disease spots spread to the entire fruit, white or grey hyphae appeared, and the fruit rotted. Twenty diseased fruits were surface-disinfested with 2% sodium hypochlorite and 75% ethanol, then rinsed with sterile distilled water to isolate the pathogen. Diseased tissue sections (5 × 3 mm) were incubated on potato dextrose agar (PDA) for 7 days at 25°C. Twelve of 15 monoconidial isolates have similar morphology. On PDA, the isolates produced copious white aerial mycelia. After 5-7 days on straw juice medium, two types of conidia appeared (Rice straw 60 g, Agar 20 g, distilled water 1000 mL) (Figure 1E-I). Macroconidia were abundant, falcate, slender, and slightly curved with 0-8 septa, mostly 4-5 septa (average 41.70 × 3.81 m, n=100) (Figure 1J). Microconidia were globose, oval, or piriform with 0-1 septa, 2.72 to 8.57 × 2.53 to 7.47 m (average 5.49 × 4.52 m, n=50) (Figure 1L), and chlamydospores were not observed. Conidial and colony morphology identified 12 monoconidial isolates as Fusarium graminearum (Fisher et al., 1982; Yulfo-Soto et al., 2021). Genomic DNA was extracted from three isolates using a DNA Extraction Kit (Yeasen, Shanghai, China). The ITS1/4 region combined with partial gene fragments of translation elongation factor-1 alpha (TEF-1α, primer TEF1/2, O'Donnell et al. 1998), RNA polymerase second largest subunit (RPB2, primer fRPB2-5F/7cR, Liu et al. 1999) and ß-tubulin (ß-tub, primer Bt2a/2b, Li et al. 2013) from the isolates were amplified and sequenced. The three tested isolates showed identical gene sequences. Sequences amplified from one representative isolate (PG16) have been submitted to GenBank. BLAST searches revealed that ITS (OM019317), TEF-1α (OM048103), RPB2 (ON364348), and ß-tub (OM048104) had 99 to 100% identity compared with F. graminearum (MH591453.1, KX087136.1, MF662636.1, and MZ078952.1, respectively) in GenBank. The phylogenetic analysis combined ITS - TEF-1α - RPB2 (O'Donnell et al. 2015) concatenated sequences using MEGA7.0 (Mao et al. 2021) showed the isolate was clustered with the F. graminearum clade with 100% bootstrap support (Figure 2). The isolate PG16 was used for pathogenicity tests. Ponkan fruits were surface-disinfested with 75% ethanol and rinsed with sterile distilled water three times. Then, 30 punctured wound fruits (2-mm-diameter, 2-mm-depth) with a sterile needle and 30 unwounded fruits were inoculated with conidial suspension (10 µL, 3.0 × 105 conidia/mL). while the control fruits were inoculated with 10 µL sterile distilled water. All fruits were incubated at 25°C and 90% relative humidity. Two days later, all wounded fruits inoculated with conidial suspension showed disease spots, similar symptoms to the original rotten fruits (Figure 1D). Control and conidial-inoculated unwounded fruits were healthy (Figure 1B-C). The Pathogenicity test was repeated twice, and similar symptoms were observed. Morphologically and molecularly, the re-isolated fungus matched the inoculated isolate. First report of F. graminearum causing Ponkan fruit rot in China. As Ponkan is an important citrus crop with high economic value in China, identification of the causing agent, F. graminearum, for fruit rot allows the development of control measures to manage this disease.

QTL Mapping of Soybean (Glycine max) Vine Growth Habit Trait.

The vine growth habit (VGH) is a notable property of wild soybean plants that also holds a high degree of importance in domestication as it can preclude using these wild cultivars for breeding and improving domesticated soybeans. Here, a bulked segregant analysis (BSA) approach was employed to study the genetic etiology of the VGH in soybean plants by integrating linkage mapping and population sequencing approaches. To develop a recombinant inbred line (RIL) population, the cultivated Zhongdou41 (ZD41) soybean cultivar was bred with ZYD02787, a wild soybean accession. The VGH status of each line in the resultant population was assessed, ultimately leading to the identification of six and nine QTLs from the BSA sequencing of the F4 population and F6-F8 population sequence mapping, respectively. One QTL shared across these analyzed generations was detected on chromosome 19. Three other QTLs detected by BSA-seq were validated and localized to the 90.93 kb, 2.9 Mb, and 602.08 kb regions of chromosomes 6 and 13, harboring 14, 53, and 4 genes, respectively. Three consistent VGH-related QTLs located on chromosomes 2 and 19 were detected in a minimum of three environments, while an additional six loci on chromosomes 2, 10, 13, and 18 were detected in at least two environments via ICIM mapping. Of all the detected loci, five had been reported previously whereas seven represent novel QTLs. Together, these data offer new insights into the genetic basis of the VGH in soybean plants, providing a rational basis to inform the use of wild accessions in future breeding efforts.

First report of Fusarium tricinctum causing fruit rot on navel orange (Citrus sinensis (L.) Osbeck) in China.

Citrus sinensis (L.) Osbeck is popular with consumers for its delicious taste. In December 2020, a rot symptom causing about 15% losses of a total of 450 fruits was observed on 'Newhall' navel oranges after 70 d storage (20℃, 85%-90% RH) at Jiangxi Key Laboratory for Postharvest Technology and Nondestructive Testing of Fruits & Vegetables (28.68° N, 115.85° E). The fruits were harvested from an orchard in Ganzhou City, Jiangxi Province, China (25.53° N, 114.79° E). Incipiently, the pedicles of infected fruits were brown, the peels became softened and showed yellowish-brown lesions which, gradually expanded and had white hyphae (Fig. S1A). To isolate the pathogen, the surface of diseased fruits was disinfected with NaClO (2%) for 2 min and ethanol (75%) for 0.5 min, then washed with sterile water three times. Tissues (5 × 5 mm) around the lesion were incubated on potato dextrose agar (PDA) at 28 ± 1℃ (L: D=12: 12) for 5 days. Five cultures with similar morphology were obtained and colonies initially produced white aerial hyphae and became khaki then turned pink on PDA (Fig. S1F, G, H). Abundant microconidia, macroconidia and rare chlamydospores were observed after 10 days on PDA and no glucose PDA media (Zhang et al. 2020). Macroconidia were falciform and curved to lunate, 2-4 septa, 29.38 × 3.75 µm in size (n=50) (Fig. S1K, Fig. S3). Microconidia were oval, napiform or pyriform, 0-1 septa, 12.00 × 3.43 µm in size (n=50) (Fig. S1L1 to L4, Fig. S3). Chlamydospores were found in hyphae, ellipsoidal or orbicular (Fig. S1I-1 to I-2, J-1 to J-2). The morphological features of five isolates were similar to Fusarium (Leslie and Summerell 2006). Genomic DNA of five isolates was extracted with DNA Extraction Kit (Yeasen, Shanghai, China), ITS1/ITS4, EF1Ha/EF2Tb and fRPB2-5F/fRPB2-7cR primers were used to amplify the internal transcribed spacer region (ITS), and the transcriptional elongation factor-1 alpha (TEF-1α), and RNA polymerase II (RPB2) gene sequences (White et al. 1990; Carbone and Kohn 1999; Liu et al. 1999). The ITS, TEF-1α and RPB2 sequences of five isolates were deposited in GenBank and showed 99-100% identity with corresponding sequences from F. tricinctum (Table S1). A phylogenetic tree was constructed with ITS-TEF-1α-RPB2 concatenated sequences in MEGA7.0 (Li et al. 2021) and all five isolates were placed in F. tricinctum clade with 100% bootstrap support (Fig. S2). To confirm pathogenicity, ten healthy C. sinensis fruits were surface-sterilized with 75% ethanol and inoculated with 10 µL spore suspension (1.0 × 106 spore/mL) including five wounded (with sterilized needle) and five unwounded (Fig. S1B to E). Control fruits were inoculated with 10 µL sterile water. All fruits were incubated at 28 ± 1℃, 90% RH for 7 days. The experiment was conducted three times. The lesion diameter of inoculated wounded fruits was 21.01 ± 2.52 mm and showed similar symptoms to original rotten fruits. However, the control and unwounded fruits remained healthy. To fulfill Koch's postulates, F. tricinctum was re-isolated from the inoculated fruits and deposited in Collaborative Innovation Center of Postharvest Key Technology and Quality Safety of Fruits and Vegetables in Jiangxi Province. To our knowledge, F. tricinctum has been reported on apple tree and kiwi plant in China (Zhang et al. 2021; Ma et al. 2022), but this is the first report of F. tricinctum causing fruit rot on navel orange in China. This finding provides important information for preventing postharvest disease of citrus.

New insights on the function of plant acyl carrier proteins from comparative and evolutionary analysis.

Acyl carrier proteins (ACPs) play a central role in both plastidial and mitochondrial Type II fatty acid synthesis in plant cells. However, a large proportion of plant ACPs remain functionally uncharacterized, and their evolutionary history remains elusive. In present study, 97 putative ACPs were identified from ten angiosperm species examined. Based on phylogenetic analysis, ACP genes were grouped into plastidial (cpACP: ACP1/2/3/4/5) and mitochondrial (mtACP: mtACP1/mtACP2/mtACP3) ACPs. Protein sequence (motifs and length), tertiary structure, and gene structure (exon number, average intron length, and intron phase) were highly conserved in different ACP subclades. The differentiation of ACPs into distinct types occurred 85-98 and 45-57 million years ago. A limited proportion of ACP genes experience tandem or segmental duplication, corresponding to two rounds of whole genome duplication. Ka/Ks ratios revealed that duplicated ACP genes underwent a purifying selection. Regarding expression patterns, most ACPs were expressed constitutively and tissue-specifically. Notably, the average expression levels of ACP1, mtACP3, and mtACP1 were positively correlated with those of ACP3, ACP4, and mtACP2, respectively. Analysis of cis-elements showed that seven motifs (CACTFTPPCA1, DOFCOREZM, GT1CONSENSUS, CAATBOX1, ARR1AT, POLLEN1LELAT52, and GATABOX) related to tissue-specific, ABA, and light-mediated gene regulation were ubiquitous in all ACPs investigated, which shed new light on the regulation patterns of these central enzymatic partners of the FAS system. This study presents a thorough overview of angiosperm ACP gene families and provides informative clues for the functional characterization of plant ACPs in the future.

LINC00992 contributes to the oncogenic phenotypes in prostate cancer via targeting miR-3935 and augmenting GOLM1 expression.

BACKGROUND: Accumulating evidence has revealed the critical role of long non-coding RNAs (lncRNAs) in cellular processes during tumor progression. As documented in cancer-related literatures, LINC00992 expression is associated with cancer progression, whereas its function in tumors including prostate cancer has not been characterized yet. METHODS: Data from GEPIA database suggested LINC00992 expression in prostate cancer tissues. The expression levels of RNAs were monitored via qRT-PCR. Western blot evaluated the levels of proteins. The proliferation, apoptosis and migration of prostate cancer cells were assessed by CCK-8, EdU, TUNEL, Transwell and wound healing assays. Luciferase reporter, RNA pull down and RIP assays were applied to detect the interplays among LINC00992, miR-3935 and GOLM1. RESULTS: Elevated levels of LINC00992 and GOLM1 were detected in prostate cancer tissues and cells. LINC00992 exerted facilitating functions in prostate cancer cell proliferation and migration. Mechanically, LINC00992 interacted with and negatively regulated miR-3935 to elevate GOLM1 expression in prostate cancer cells. In addition, the in vitro suppressive effect of silenced LINC00992 on prostate cancer cell proliferation and migration was reversed by GOLM1 upregulation. Likewise, LINC00992 depletion restrained tumor growth in vivo was offset by enhanced GOLM1 expression. CONCLUSIONS: LINC00992 competitively bound with miR-3935 to elevate GOLM1 expression and therefore facilitate the oncogenic phenotypes of prostate cancer cells, implying a potential LINC00992-targeted therapy for prostate cancer.

[Acute oral toxicity and 90 days feeding test of recombinant human lactoferrin].

OBJECTIVE: To study the edible safety of recombinant human lactoferrin( rh LF) expressed from transgenic cow mammary gland bioreactor. METHODS: According to the food additive safety toxicology evaluation procedures and method, acute oral toxicity in rats and 90 day subchronic toxicity test in mice were done to evaluate the edible safety of rh LF. RESULTS: Acute oral toxicity test indicated that rh LF was no toxic effect during the observation period, mouse acute oral LD50 of recombinant human lactoferrin was greater than 20 000 mg/kg. 90 days feeding test indicated that there was no-observedadverse-effect-level after givening 300 times rh LF recommended dose of animals body, toxicological parameters NOAEL was 10. 00 g/( kg·d). CONCLUSION: According to the acute toxic dose graduation standard, rh LF was nonpoisonous. Rh LF was no-observedadverse-effect-level and no subchronic toxicity after givening 300 times rh LF recommended dose of animals body. According to the result, rh LF was no potential food safety risk.

Prediction of Fuhrman pathological grade of renal clear cell carcinoma based on CT texture analysis.

OBJECTIVE: To study the predictive performance of the imaging model based on the texture analysis of CT plain scan in distinguishing between low (grade I and II) and high (grade III and IV) of Fuhrman pathological grade of renal clear cell carcinoma. METHODS: The clinical data of 94 patients with ccRCC who underwent CT scan and were confirmed by biopsy or surgery in TCGA-KIRC public database were retrospectively analyzed. There were 32 cases of low-grade ccRCC and 62 cases of high-grade ccRCC. The patients were randomly divided into training set and verification set according to the proportion of 7:3 by stratified sampling method. The imaging characteristics of ccRCC were calculated in the plain CT images. Lasso regression was used to reduce the dimensionality of the imaging characteristics of the training set, and binary logistic regression was used to construct the prediction model. Bootstrap method was used to verify the training set model and the validation set model, and the area under the receiver operating characteristic (ROC) curve (AUC) was calculated respectively. RESULTS: Binary logistic regression showed that only imaging features were independent risk factors for predicting the Furhman classification of ccRCC. The predictive model was y = 1/[1 + exp (-z)], z = 1.274 × imaging risk score + 0.072. The results of bootstrap internal validation showed that the AUC of the training group was 0.961 (95% CI: 0.900-0.913). The Hosmer-Lemeshow goodness of fit test showed that the prediction model had a good calibration in the training group (P = 0.416). The AUC of prediction model in validation group was 0.731 (95% CI: 0.500-1.000). The Hosmer-Lemeshow goodness of fit test results showed that the prediction model had a good calibration in the validation group (P = 0.592). CONCLUSION: The model based on CT texture analysis has a good predictive effect in differentiating low-grade and high-grade ccRCC and can provide reference for the treatment and prognosis of patients.

Social Support and Physical Activity in College and University Students: A Meta-Analysis.

Although physical activity (PA) has a profound impact on health, many college and university students are still physically inactive. There is some evidence to suggest that social support (SS) could impact the PA levels of students, but the internal relationship and specific effects are not very clear. The purpose of this review was to determine the strength of the relationship between SS and PA and examine whether any potential associations differed in terms of age, gender, and region among college and university students. Studies were identified using the following electronic databases: PubMed, SPORTDiscus, Web of Science, and Sociological Abstracts. Moderator analyses investigating the effects of students' age, gender, and region (nation) were performed. This review included 19 articles. The results showed total SS was significantly associated with PA (r = 0.30, 95% confidence interval [CI]: [0.22, 0.37], p < .001). With respect to different types of support, friend support was more strongly associated with PA than family support. Gender factors had a significant moderating effect on the correlation between SS and PA (QM = 17.433, p < .001). Separate analyses examining the moderating effects of gender (percentage of females) found that the association between SS and PA was stronger with the increase in female percentage. In conclusion, SS is an important factor associated with PA levels and should foster SS within intervention programs according to types of SS and gender differences to increase PA levels among college and university students.

Improving maize carbon and nitrogen metabolic pathways and yield with nitrogen application rate and nitrogen forms.

Reduced fertilizer efficiency caused by excessive use of nitrogen (N) fertilizer is a major problem in agriculture and a hot topic of research. Most studies have focused on the effect of N application rate on N efficiency, whereas there are limited studies on changing the N form to improve N yield and efficiency. Here, the effects of different N application rates and nitrate-to-ammonium N ratios on post-anthesis carbon (C) and N metabolism and maize yield under shallow-buried drip irrigation were investigated. Two rates of N application (210 kg·ha-1 (NA1) and 300 kg·ha-1 (NA2)) and three nitrate-to-ammonium N ratios (2:1 (NF1), 3:1 (NF2), and 4:1 (NF3)) were utilized. Post-anthesis photosynthetic characteristics, activities of key enzymes in photosynthetic C and N metabolism, nonstructural carbohydrate content, post-anthesis N accumulation and transportation, yield, and N-use efficiency were determined. At both N application rates, NF2 treatment enhanced photosynthetic activity in the ear-leaf at silking stage and promoted key enzyme activities of C and N metabolic pathways, compared with NF1 and NF3. Furthermore, NF2 significantly increased nonstructural carbohydrate accumulation (4.00-64.71%), post-anthesis N accumulation and transportation (11.00-38.00%), and grain yield (2.60-13.08%). No significant differences between NA1 and NA2 were observed under NF2 in most of the measured variables; however, NA1 had higher N-use efficiency. Thus, the optimal treatment under shallow-buried drip irrigation was a N application rate of 210 kg ha-1 and a nitrate-to-ammonium N ratio of 3:1. These findings provide theoretical guidance on appropriate N applications for high-yield maize production.

Metabolomic profiling reveals biomarkers for diverse flesh colors in jelly fungi (Auricularia cornea).

Auricularia cornea has garnered attention due to its nutrition, culinary applications, and promising commercial prospects. However, there is little information available regarding the metabolic profiling of various colors strains. In this study, 642 metabolites across 64 classes were identified by LC-MS/MS to understand the metabolic variations between white, pink and dark brown strains. Notably, prenol lipids, carboxylic acids and fatty acyls accounted for 46.8 % of the total. Comparative analysis revealed 17 shared differential metabolites (DMs) among them. ACP vs ACW exhibited 17 unique metabolites, including d-arginine and maleic acid, etc. ACP vs ACB showed 5 unique metabolites, with only PS(18:1(9Z)/0:0) demonstrating up-regulation. ACB vs ACW showed 8 unique metabolites, including 4-hydroxymandelic acid and 5'-methylthioadenosine, etc. KEGG enrichment analysis highlighted pathway variations, and MetPA analysis identified key-pathways influencing DMs accumulation in A. cornea. This pioneering metabolomics study offers insights into A. cornea metabolic profiling, potential applications, and guides further research.

Research progress on surgical factors related to early urinary control after laparoscopic radical prostatectomy.

Radical resection of prostate cancer is the first choice for the treatment of early localized prostate cancer, but urinary incontinence is prone to occur after the operation, especially early urinary incontinence, which seriously affects the quality of life of patients. This article discusses the surgical methods, approaches, and techniques to clarify the effects of surgical-related factors on early postoperative urinary control, in order to provide the best treatment for patients with prostate cancer.

Clinical study of 3D laparoscopic radical prostatectomy by transperitoneal and extraperitoneal approaches.

OBJECTIVE: Comparison of the clinical effectiveness and safety of three-dimensional transperitoneal laparoscopic radical prostatectomy (3D TLRP) versus 3D extraperitoneal LRP (3D ELRP) for prostate cancer. MATERIALS AND METHODS: To retrospectively analyze the clinical and regular postoperative follow-up data of patients who underwent 3D LRP performed by the same attending surgeon at the Affiliated Hospital of Bengbu Medical College between 2017 and 2022. A total of 82 patients who met the criteria were included. They were divided into 3D TLRP (n = 39) and 3D ELRP groups (n = 43) according to the surgical approach. The preoperative, intraoperative, and postoperative data were compared. RESULTS: There were no statistically significant differences in preoperative characteristics between the two groups. There were also no statistically significant differences between the 3D TLRP and 3D ELRP groups in terms of intraoperative blood transfusion rate (12.82% vs. 2.33%), positive lymph node rate (11.11% vs. 2.38%), positive surgical margin rate (12.82% vs. 6.98%), pathological Gleason score, postoperative clinical stage, perioperative complication rate (10.26% vs. 4.65%), immediate urinary control rate (56.41% vs. 58.14%), 3-month postoperative urinary control rate (76.92% vs. 74.42%), 6-month postoperative urinary control rate (87.18% vs. 83.72%), 6-month postoperative biochemical recurrence rate (7.69% vs. 9.30%), or 6-month postoperative sexual function recovery rate (2.56% vs. 2.33%) (P > 0.05). Compared with the 3D ELRP group, the 3D TLRP group had a longer operative time (232.36 ± 48.52 min vs. 212.07 ± 41.76 min), more estimated blood loss (150.000 [100.0, 200.0] vs. 100.000 [100.0, 125.0]), longer recovery of gastrointestinal function (2.72 ± 0.89 vs. 2.26 ± 0.88), longer duration of drainage tube retention (5.69 ± 1.79 vs. 4.28 ± 2.68), and longer hospitalization time (12.54 ± 4.07 vs. 10.88 ± 2.97), with statistical significance (P < 0.05). CONCLUSION: 3D TLRP and 3D ELRP have similar oncologic and functional outcomes. Clinically, physicians can choose a reasonable procedure according to the patient's specific situation and their own surgical experience.

Pathogenic invasive microbes Trichoderma pleuroticola transform bacterial and fungal community diversity in Auricularia cornea crop production system.

Pathogenic invasion of Trichoderma pleuroticola profoundly altered microflora in the Auricularia cornea crop production system, impacting diversity and composition in both artificial bed-log and fruiting bodies. A more complex ecological network between the diseased and healthy bodies. Researchers still have poor knowledge about how the important agricultural relationship between the composition of the microbiome of the artificial bed-log and the fruiting bodies is infected by the pathogenic invasive microbes T. pleuroticola, but this knowledge is crucial if we want to use or improve it. Here, we investigated 8 groups (48 biological samples) across 5 growth stages of the A. cornea production system using metagenomic technology. Diseased and healthy fruiting bodies exhibited distinct microbial compositions, while core members in artificial bed-logs remained stable. Core microbiota analysis highlighted Pseudomonas and Pandoraea bacterial genera, as well as Sarocladium, Cephalotrichum, Aspergillus, and Mortierella fungal genera as biomarker species after the bodies were treated with the pathogenic invasive microbes T. pleuroticola. In diseased bodies, these core members upregulated pathways including polymyxin resistance, L-arginine degradation II, superpathway of L-arginine and L-ornithine degradation, glucose degradation (oxidative), glucose and glucose-1-phosphate degradation, promoting fruit spoilage. Our data confirm that T. pleuroticola plays an important role in the early stages of disease development in the A. cornea crop generation system. The exposed volatile core microbiome may play an important role in accelerating T. pleuroticola-induced decay of fruiting bodies.

Effects of blood flow restriction training on bone metabolism: a systematic review and meta-analysis.

Introduction: The efficacy of low-intensity blood flow restriction (LI-BFR) training programs in bone metabolism remains unclear compared to low-intensity (LI) training and high-intensity (HI) training. The aim of this review was to quantitatively identify the effects of LI-BFR training on changes in bone formation markers (i.e., bone-specific alkaline phosphatase, BALP), bone resorption (i.e., C-terminal telopeptide of type I collagen, CTX) and bone mineral density (BMD) compared with conventional resistance training programmes. Additionally, the effectiveness of walking with and without BFR was assessed. Methods: PubMed, Scopus, SPORTDiscus, Web of Science and Google Scholar databases were searched for articles based on eligibility criteria. Review Manager Version 5.4 was used for Meta-analysis. Physiotherapy Evidence Database (PEDro) was applied to assess the methodological quality of studies. Results: 12 articles were included in the meta-analysis, with a total of 378 participants. Meta-results showed that compared with LI training, LI-BFR training induced greater increments in BALP (young adults: MD = 6.70, p < 0.001; old adults: MD = 3.94, p = 0.002), slight increments in BMD (young adults: MD = 0.05, p < 0.00001; old adults: MD = 0.01, p < 0.00001), and greater decrements in CTX (young adults: MD = -0.19, p = 0.15; old adults: MD = -0.07, p = 0.003). Compared with HI training, LI-BFR training produced smaller increments in BALP (young adults: MD = -6.87, p = 0.24; old adults: MD = -0.6, p = 0.58), similar increments in BMD (MD = -0.01, p = 0.76) and similar decrements in CTX (young adults: MD = 0, p = 0.96; old adults: MD = -0.08, p = 0.13). Although there were only two studies on walking training intervention, walking training with BFR had a better effect on bone metabolism than training without BFR. Discussion: In conclusion, LI-BFR training induces greater improvements in bone health than LI training, but is less effective than HI training. Therefore, LI-BFR training may be an effective and efficient way to improve bone health for untrained individuals, older adults, or those undergoing musculoskeletal rehabilitation. Clinical Trial Registration: [https://www.crd.york.ac.uk/prospero/], identifier [CRD42023411837].

A microbial ecosystem enhanced by regulating soil carbon and nitrogen balance using biochar and nitrogen fertiliser five years after application.

The indiscriminate use of nitrogen fertiliser (NF) is a obstruction to improve soil quality and crop yields. However, the effect of biochar and NF on soil microbial ecosystem (SME) and crop yields is unknown. A five-year field experiment in China aimed to evaluate the effects of biochar and nitrogen fertiliser (NF) combination on soil structure, C-to-N ratio (CNR), microbial biomass, and spring maize yield. Biochar and NF were applied at different rates, and the combined application resulted in a soil solid-liquid-gas ratio closer to the ideal value. The use of biochar alone and in combination with NF significantly increased soil's C, N, and CNR. A moderate application of biochar and NF resulted in favourable biological and chemical properties of the soil. The application of biochar and NF at moderate levels led to an increase in SME, with the B8N150 producing the highest yield. The highest yield of B8N150 represents a 24.25% increase compared to the unfertilized control and a 9.04% increase compared to B0N150. Moderate use of biochar and NF could be beneficial in areas with similar climatic conditions.

Scrotal calcinosis: a case report and literature review.

OBJECTIVE: To understand the clinical features of idiopathic scrotal calcium deposits, to improve the understanding of the disease, and to discuss its etiology, pathogenesis and scrotal reconstruction strategies in the course of diagnosis and treatment. METHODS: To analyze the diagnosis and treatment of one case of idiopathic calcium salt deposition in the scrotum and to review the relevant literature. RESULTS: The patient was a 55-year-old male with multiple yellowish-white nodules of varying sizes in the scrotum for more than 20 years, with hard nodules and no tenderness or ulcerative manifestations. Under subarachnoid anesthesia, the scrotum was reconstructed after surgical excision of all diseased nodes, and postoperative pathology was consistent with scrotal calcium salt deposition. CONCLUSION: Scrotal idiopathic calcium deposits is a rare skin conditions caused by insoluble calcium salts deposited in the scrotal skin tissue, for which surgical excision of the lesion is the main treatment modality with remarkable results. It needs to be differentiated from epidermoid cysts, multiple lipodystrophies of the scrotum, and scabies nodules.