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Retinal fibrosis is one of the major features of diabetic retinopathy (DR). Our recent research has shown that Poldip2 can affect early DR through oxidative stress, but whether Poldip2 would regulate retinal fibrosis during DR development is still enigmatic. Here, diabetic Sprague-Dawley (SD) rats were induced with streptozotocin (STZ) and treated with adeno-associated virus serotype 9–polymerase-δ interacting protein 2 (Poldip2) shRNA, while human adult retinal pigment epithelial (ARPE-19) cells were treated with high glucose or Poldip2 siRNA. We identified that in STZ-induced DR rats and ARPE-19 cells treated with high glucose, the expression of Poldip2, transforming growth factor-β1 (TGF-β1), phosphorylated-SMAD3/SMAD3, MMP9, COL-1, FN, and CTGF increased while the expression of cadherin decreased. However, deleting Poldip2 inhibited the TGF-β1/SMAD3 signaling pathway and attenuated the above protein expression in vivo and in vitro. Mechanistically, we found that Poldip2 promotes the activation of SMAD3, facilitates its nuclear translocation through interacting with it, and significantly enhances the expression of fibrosis makers. Collectively, Poldip2 was identified is a novel regulator of DR fibrosis and is expected to become a therapeutic target for PDR. Article Highlights: To investigate the role of polymerase-δ interacting protein 2 (Poldip2) in retinal fibrosis during diabetic retinopathy (DR) development. Does Poldip2 regulate retinal fibrosis in DR, and what are the underlying mechanisms? Poldip2 upregulation in DR correlates with increased expression of fibrotic markers, while Poldip2 deletion inhibits the transforming growth factor-β1/SMAD3 pathway, reducing fibrosis-related protein levels. Poldip2 emerges as a novel regulator of DR fibrosis, suggesting therapeutic potential for treating proliferative diabetic retinopathy. [ABSTRACT FROM AUTHOR]

Background: Diabetic nephropathy (DN) is a severe complication of diabetes influenced by arginine methylation. This study aimed to elucidate the role of protein arginine methylation-related genes (PRMT-RGs) in DN and identify potential biomarkers. Methods: Differentially expressed genes in two GEO datasets (GSE30122 and GSE104954) were integrated with 9 PRMT-RGs. Candidate genes were identified using WGCNA and differential expression analysis, then screened using support vector machine-recursive feature elimination and least absolute shrinkage and selection operator. Biomarkers were defined as genes with consistent differential expression across both datasets. Regulatory networks were constructed using the miRNet and Network Analyst databases. Gene set enrichment analysis was performed to identify the signaling pathways in which the biomarkers were enriched in DN. Different immune cells in DN were identified using immune infiltration analysis. Meanwhile, drug prediction and molecular docking identified potential DN therapies. Finally, qRT-PCR and immunohistochemistry validated two biomarkers in STZ-induced DN mice and DN patients. Results: Two biomarkers (FAM98A and FAM13B) of DN were identified in this study. The molecular regulatory network revealed that FAM98A and FAM13B were co-regulated by 6 microRNAs and 1 transcription factor and were enriched in signaling pathways. Immune infiltration and correlation analyses revealed that FAM98A and FAM13B were involved in developing DN along with PRMT-RGs and immune cells. The expression levels of Fam98a and Fam13b were significantly upregulated in the kidneys of DN mice revealed by qRT-PCR analysis. The expression levels of FAM98A were significantly upregulated in the kidneys of DN patients revealed by immunohistochemistry staining. Molecular docking showed that estradiol and rotenone exerted potential therapeutic effects on DN by targeting FAM98A. Conclusion: Comprehensive bioinformatics analysis revealed that FAM98A and FAM13B were potential DN biomarkers correlated with PRMT-RGs and immune cells. This study provided useful insights for elucidating the molecular mechanisms and developing targeted therapy for DN. [ABSTRACT FROM AUTHOR]

Motivation Human epidermal growth factor receptor 2 (HER2) status identification enables physicians to assess the prognosis risk and determine the treatment schedule for patients. In clinical practice, pathological slides serve as the gold standard, offering morphological information on cellular structure and tumoral regions. Computational analysis of pathological images has the potential to discover morphological patterns associated with HER2 molecular targets and achieve precise status prediction. However, pathological images are typically equipped with high-resolution attributes, and HER2 expression in breast cancer (BC) images often manifests the intratumoral heterogeneity. Results We present a phenotype-informed weakly supervised multiple instance learning architecture (PhiHER2) for the prediction of the HER2 status from pathological images of BC. Specifically, a hierarchical prototype clustering module is designed to identify representative phenotypes across whole slide images. These phenotype embeddings are then integrated into a cross-attention module, enhancing feature interaction and aggregation on instances. This yields a phenotype-based feature space that leverages the intratumoral morphological heterogeneity for HER2 status prediction. Extensive results demonstrate that PhiHER2 captures a better WSI-level representation by the typical phenotype guidance and significantly outperforms existing methods on real-world datasets. Additionally, interpretability analyses of both phenotypes and WSIs provide explicit insights into the heterogeneity of morphological patterns associated with molecular HER2 status. Availability and implementation Our model is available at https://github.com/lyotvincent/PhiHER2 [ABSTRACT FROM AUTHOR]

Panax notoginseng is a perennial plant well known for its versatile medicinal properties, including hepatoprotective, antioxidant, anti-inflammatory, anti-tumor, estrogen-like, and antidepressant characteristics. It has been reported that plant age affects the quality of P. notoginseng. This study aimed to explore the differential metabolome and transcriptome of 2-year (PN2) and 3-year-old (PN3) P. notoginseng plant root samples. Principal component analysis of metabolome and transcriptome data revealed major differences between the two groups (PN2 vs. PN3). A total of 1813 metabolites and 28,587 genes were detected in this study, of which 255 metabolites and 3141 genes were found to be differential (p < 0.05) between PN2 vs. PN3, respectively. Among differential metabolites and genes, 155 metabolites and 1217 genes were up-regulated, while 100 metabolites and 1924 genes were down-regulated. The KEGG pathway analysis revealed differentially enriched metabolites belonging to class lipids ("13S-hydroperoxy-9Z, 11E-octadecadionic acid", "9S-hydroxy-10E, 12Z-octadecadionic acid", "9S-oxo-10E, 12Z-octadecadionic acid", and "9,10,13-trihydroxy-11-octadecadionic acid"), nucleotides and derivatives (guanine and cytidine), and phenolic acids (chlorogenic acid) were found to be enriched (p < 0.05) in PN3 compared to PN2. Further, these differentially enriched metabolites were found to be significantly (p < 0.05) regulated via linoleic acid metabolism, nucleotide metabolism, plant hormone signal transduction, and arachidonic acid metabolism pathways. Furthermore, the transcriptome analysis showed the up-regulation of key genes MAT, DMAS, SDH, gallate 1-beta-glucosyltransferase, and beta-D-glucosidase in various plants' secondary metabolic pathways and SAUR, GID1, PP2C, ETR, CTR1, EBF1/2, and ERF1/2 genes observed in phytohormone signal transduction pathway that is involved in plant growth and development, and protection against the various stressors. This study concluded that the roots of a 3-year-old P. notoginseng plant have better metabolome and transcriptome profiles compared to a 2-year-old plant with importantly enriched metabolites and genes in pathways related to metabolism, plant hormone signal transduction, and various biological processes. These findings provide insights into the plant's dynamic biochemical and molecular changes during its growth that have several implications regarding its therapeutic use. [ABSTRACT FROM AUTHOR]

Mountain-cultivated ginseng ( MCG) can be considered a mimic of wild ginseng, whose seeds are sowed artificially, cultivated in the natural environment, and returned to the wild state before being used clinically. Cultivated ginseng ( CG) and MCG are mainly used as the commercial material for clinical applications. However, MCG is much more expensive but effective than CG. The aim of this study is to explore the differences in the pharmacokinetics and brain concentration of Rg1, Re, Rb1, and Rd after oral administration of MCG, CG, and pure ginsenosides. An ultra-performance liquid chromatography-tandem mass spectrometry method was developed and validated to determine the concentration of the four ginsenosides in rat plasma and brain tissue. Compared with the pure group, the area under the curve ( AUC) of all the four ginsenosides for CG and MCG increased. The mean brain concentrations of the four ginsenosides were found to be 10- to 15-fold lower than the corresponding contents in the plasma, and the poor permeability of ginsenosides into blood-brain barrier was indicated by the low concentrations of the four ginsenosides. [ABSTRACT FROM AUTHOR]

• Ascomycota and Basidiomycota are the dominant phyla in Panax ginseng -growing soil. • Changes induced by all treatments on soil fungal communities are evaluated. • Adding different leaf litter alters the composition of fungal community. • Basidiomycota were negatively affected by total nitrogen. • Some active biomarkers may be involved in the decomposition of different leaf litters. Soil fungi communities play a vital role in the plant-soil ecosystem and affect plant growth and health, but the underlying mechanism controlling soil fungi communities in understory wild ginseng soil is still unknown. To study that mechanism, a pot culture experiment adding different leaf litters based on a completely randomized design was carried out. The results indicated that 1990 operational taxonomic units were obtained from eighteen samples. Ascomycota and Basidiomycota, the two dominant phyla, accounted for 84.57% of the total valid reads at the phylum level. The observed species and Chao 1 index in treatment E were significantly lower than those in the other treatments, although no significant differences were found among the Shannon index of all treatments (p > 0.05). Different types of leaf litter significantly changed the fungal community composition; specifically, Ascomycota was higher in broad leaf litter treatments (A, C, D and E) than in the coniferous leaf litter treatment (B), but Basidiomycota showed the opposite trend, indicating that Ascomycota and Basidiomycota could be used to identify the species of coniferous and broad leaves. The active biomarker fungal (Bmf), 36 different phylotypes, were identified by a linear discriminant analysis effect size algorithm in all treatments, and some Bmf may participate in the decomposition of different tree litters. Additionally, the changes in fungal community diversity and composition were closely related to the changes in soil microbial biomass carbon, total nitrogen and available phosphorus in all treatment soils. Overall, our study indicated that leaf litter changed the soil fungal community structure, and some Bmf were directly involved in leaf litter decomposition. Bmf is more indicative and helpful for distinguishing leaf species. [ABSTRACT FROM AUTHOR]

Abstract Rusty root is a widespread problem in ginseng production, which can reduce ginseng quality and root price. The formation of rusty root is a complex process caused by multiple factors, primarily soil physicochemical factors and microorganisms. However, the precise roles played by these factors in the development of this disease remain unclear, especially the role of microorganisms. Deciphering the structure of the rhizosphere microbial communities will provide a more comprehensive and deeper understanding of rusty root disease. Rhizosphere microbial communities from five ginseng fields with different rusty root grades and indices were characterized by MiSeq sequencing. The pattern of beta diversity indicated that rusty root disease might be positively correlated with bacterial community structure, but not with fungal community structure. Rusty root may be caused by excessive Fe (II) oxidation and Fe (III) deposition accompanied by aluminum (Al) and manganese (Mn), in which nitrate-dependent Fe (II)-oxidizing bacteria, including Acidobacteria, Chloroflexi, and Nitrospirae, play a driving role. Moreover, the accumulation of iron (Fe), Al, and Mn could enhance growth and weak invasion of some fungi, thus exacerbating the rust symptoms. However, no correlation was found between the relative abundance of the genus Ilyonectria and rusty root grade or index (P > 0.05); suggesting that Ilyonectria species may not be essential for rusty root disease. Thus, the oxidation and deposition of rhizosphere Fe, Al, and Mn facilitated by nitrate-dependent Fe (II)-oxidizing bacteria might be key factors contributing to rusty root disease. Further studies on the inhibition of Fe, Al, and Mn transformation and accumulation, and application of iron/nitrogen-cycling related bacteria in ginseng soil might elucidate effective methods for preventing rusty root. Highlights • Rusty root could be a physiological disorder but not an infectious disease. • Rusty root may be caused by Fe (II) oxidation and deposition. • Nitrate-dependent Fe(II) oxidizing bacteria play a driving role in rusty root. [ABSTRACT FROM AUTHOR]

[Display omitted] • Ascomycota and Basidiomycota had the same change trend in ginseng soils of all ages. • Soil harmful microbes predominate as culture time increased in ginseng. • Enhanced synergy of pathogenic bacteria with the extension of cultivation time. • Decreased amino acid synthesis and physiological metabolism in aged ginseng soils. • Soil pH was a driver of microbial community structure in cultivated ginseng soil. Changes in the microbial community are considered important factors affecting the yield and quality of ginseng cultivated in farmland, while the characteristics of the change in the microbial community of the soil before and after cultivation of ginseng in farmland are not well understood. Here, we aimed to fill this research gap by comparing the differences in soil nutrients and microbial communities between continuously cultivated ginseng soils for 1-, 2-, 3- and 5-year-old ginseng soils and corresponding uncultivated ginseng soils. The soil properties (pH, total soil organic carbon (TOC) content and total nitrogen (TN) content) of aged ginseng soil (3- and 5-year-old) were significantly reduced compared with those of the control soil after ginseng was cultivated in farmland. With the exception of 2-year-old ginseng, soil bacterial richness and diversity were higher in different ages of cultivated ginseng soils than in the control soil, while fungal diversity decreased in aged ginseng soil and increased in young ginseng soil (1- and 2-year-old). Ascomycota and Basidiomycota showed a consistent increasing/decreasing trend in cultivated ginseng soils of all ages compared to the control. In addition, ginseng cultivation resulted in a substantial decrease in the complexity of soil bacterial networks but a slight decrease in the complexity of fungal networks. The results from the linear discriminant size effect (LEfSe) analysis showed that beneficial microorganisms were biomarkers of the young ginseng soil, including Nitrosospira , Cephalotrichum and Humicola , while harmful microorganisms were biomarkers of aged ginseng, such as Plectosphaerella , Cladosporium and Alternaria. PICRUSt2 analysis revealed that the increase in the incidence of ginseng disease with continuous cultivation time may be related to the decreases in amino acid synthesis and physiological metabolism and the increase in fungal saprophytes. Soil pH was the most common and best predictor of bacterial and fungal community structure, followed by TOC and ammonium nitrogen (NH 4 +-N) contents. Meanwhile, the decreased of soil pH, TOC and AK may be the driving factors for the growth of ginseng soil pathogens. Overall, this study revealed the changing ginseng rhizosphere soil microbial community at different continuous cultivation times, provided a theoretical basis for ginseng cultivation, and promoted the development of medicinal plants such as ginseng. [ABSTRACT FROM AUTHOR]

• A minimum data set was established for soil quality assessment. • Two soil factors offer the greatest potential for monitoring soil quality. • Positive impacts on accumulation of Astragalus mongholicus have grown over time. • Positive correlation between soil quality index and effective ingredients. The Minimum Data Set (MDS) is an effective way to improve understanding of the assessing soil quality and understanding of the heterogeneity of soil quality and encourage the adaption of proper agriculture practice. However, a suitable evaluation method of the soil quality of Astragalus mongholicu s and ecological link between this and the active ingredients, including astragalosides (I, II, and IV) and calycosin, was poorly known in the main cultivation region in China. This study selected 22 soil indicators and 4 active ingredients factors and used principle component analysis to construct MDS to determine a comprehensive set of indicators for assess soil quality and established the agriculture practice in the main cultivation regions of A. mongholicus. Our results indicated that the quality of A. mongholicus, based on the quality score of the active ingredients from the PCA , increased with an increase in the soil quality. The five soil factors, soil organic matter(SOM), soil total calcium(STCa), soil total gallium(STGa), soil total borom(STB), and soil total magnesium(STMg), were obtained within 10% of the highest score of the total soil scores and r < 0.5 by the MDS method. In addition, the assessment quality index (AQI) of A. mongholicus was obtained from the score and contribution in the PCA and the soil quality index (SQI) was assessed by the MDS method. Interestingly, SOM, STCa were the limiting factors influencing the astragaloside II and IV content. Additionally, a significantly positive correlation was found between the SQI (r = 0.372, P < 0.01), pH (r = 0.260, P < 0.05) and AQI. The findings of this study provided a novel approach for the soil quality evaluation of A. mongholicus and the SOM, STCa and pH may be an effective and practical tool to achieve a strategic regional goal, with respect to a sustainable high quality of A. mongholicus. [ABSTRACT FROM AUTHOR]

The biomass production as well as the accumulation of secondary metabolites of plant is highly determined by the absorption of nutritional elements, in particular nitrogen (N) and phosphorus (P). Arbuscular mycorrhizal fungi (AMF) can absorb soluble P and transport it to plants, while phosphate solubilizing bacteria (PSB) can increase the content of solubilizing P in soil. Previous studies have identified the effects of either AMF or PSB inoculation on altering plant C:N:P stoichiometry, whether AMF interact with PSB in promoting plant growth and changing elemental concentration and composition of secondary metabolites by altering plant C:N:P stoichiometry remains ambiguous. In this study, we investigated the effects of inoculation of AMF, PSB, and their co-inoculation AMP (AMF and PSB) on the biomass growth, the C:N:P stoichiometry, the core microorganisms of rhizosphere soil, and the ginsenoside compositions of ginseng ( Panax ginseng ). The results showed that compared to control or single inoculation of AMF or PSB, co-inoculation of AMF and PSB significantly increased the AMF colonization rate on ginseng roots, increased the biomass of both above and under-ground parts of ginseng. Similarly, co-inoculation of AMF and PSB substantially increased the concentrations of N and P, reduced the ratios of C:P and N:P in the above-ground part of ginseng. The co-inoculation of AMF and PSB also increased concentrations of total ginsenosides and altered the compositions of ginsenosides in both the above and under-ground parts of ginseng. Analysis the rhizosphere microorganism showed that the co-inoculation of AMF and PSB recruited distinct core microorganisms that differ from the control and treatments with single inoculation of AMF or PSB. Our results suggested that PSB inoculation enhanced the positive effect of AMF in improving the absorption of nutrimental elements, altered the C:N:P stoichiometry and, ginsenosides concentration and composition of ginseng, influenced the plant rhizosphere microbial community. These findings offer valuable insights into enhancing plant biomass production and promoting secondary metabolites by improving the plant-fungi-bacterial relationships., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Mu, Ding, Zhang, Jin, Liu, Guan, Zhang, Liang, Zhou and Liu.)

Hemophagocytic lymphohistiocytosis (HLH) is an immune dysfunction characterized by an exaggerated and pathological inflammatory response, potentially leading to systemic inflammatory reactions and multiple-organ failure, including renal involvement. HLH can be classified as primary or secondary, with primary HLH associated with genetic mutations affecting cell degranulation capacity, and secondary HLH often linked to infections, tumors, and autoimmune diseases. The pathogenesis of HLH is not fully understood, but primary HLH is typically driven by genetic defects, whereas secondary HLH involves the activation of CD8+ T cells and macrophages, leading to the release of inflammatory cytokines and systemic inflammatory response syndrome (SIRS). The clinical presentation of HLH includes non-specific manifestations, making it challenging to differentiate from severe sepsis, particularly secondary HLH due to infections. Shared features include prolonged fever, hepatosplenomegaly, hematopenia, hepatic dysfunction, hypertriglyceridemia, and hypofibrinogenemia, along with histiocytosis and hemophagocytosis. However, distinctive markers like dual hemocytopenia, hypertriglyceridemia, hypofibrinogenemia, and elevated sCD25 levels may aid in differentiating HLH from sepsis. Indeed, no singular biomarker effectively distinguishes between hemophagocytic lymphohistiocytosis and infection. However, research on combined biomarkers provides insights into the differential diagnosis. Renal impairment is frequently encountered in both HLH and sepsis. It can result from a systemic inflammatory response triggered by an influx of inflammatory mediators, from direct damage caused by these factors, or as a consequence of the primary disease process. For instance, macrophage infiltration of the kidney can lead to structural damage affecting various renal components, precipitating disease. Presently, tubular necrosis remains the predominant form of renal involvement in HLH-associated acute kidney injury (HLH-AKI). However, histopathological changes may also encompass interstitial inflammation, glomerular abnormalities, microscopic lesions, and thrombotic microangiopathy. Treatment approaches for HLH and sepsis diverge significantly. HLH is primarily managed with repeated chemotherapy to eliminate immune-activating stimuli and suppress hypercellularity. The treatment approach for sepsis primarily focuses on anti-infective therapy and intensive symptomatic supportive care. Renal function significantly influences clinical decision-making, particularly regarding the selection of chemotherapy and antibiotic dosages, which can profoundly impact patient prognosis. Conversely, renal function recovery is a complex process influenced by factors such as disease severity, timely diagnosis, and the intensity of treatment. A crucial aspect in managing HLH-AKI is the timely diagnosis, which plays a pivotal role in reversing renal impairment and creating a therapeutic window for intervention, may have opportunity to improve patient prognosis. Understanding the clinical characteristics, underlying causes, biomarkers, immunopathogenesis, and treatment options for hemophagocytic lymphohistiocytosis associated with acute kidney injury (HLH-AKI) is crucial for improving patient prognosis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhao, Guan, Lin, Qiu, Zhao and Duan.)

Introduction: Ginseng ( Panax ginseng C.A. Meyer) has multiple effects on human health; however, soil degradation seriously affects its yield. Trichoderma spp. play an important role in improving plant biomass by influencing the soil environment. Therefore, it is necessary to screen efficient Trichoderma strains that can increase ginseng biomass and determine their mechanisms., Methods: Herein, we selected six Trichoderma species ( T. brevicompactum , T. velutinum , T. viridescens , T. atroviride , T. koningiopsis , and T. saturnisporum ) isolated from ginseng rhizosphere soil, and evaluated their growth promoting effects on ginseng and their influence on the microbiome and chemical attributes of the ginseng rhizosphere soil., Results: Except for T. saturnisporum (F), compared with the control, the other five species increased ginseng biomass. In terms of chemical properties, the pH value, available potassium content, and available phosphorus content in the ginseng rhizosphere soil increased by 1.16-5.85%, 0.16-14.03%, and 3.92-38.64%, respectively, after root irrigation with spores of Trichoderma species. For the soil microbiome, fungal Chao1 and Ace richness indices decreased. Application of Trichoderma enhanced the relative level of Proteobacteria , but reduced the relative level of Ascomycota . At the genus level, application of Trichoderma enhanced the relative levels of Sphingomonas, Blastomonas , and Trichoderma , but reduced the relative level of Fusarium . Available K and available P were the most important elements that affected the structure of the bacterial community, while total K was the most influential element for the structure of the fungal community structure., Conclusion: The results indicated that the application of Trichoderma spp. could increase soil nutrients and regulate the structure and composition of the soil microbial community, thereby enhancing the biomass of ginseng. The results will provide guidance for soil improvement in ginseng cultivation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhang, Jin, Guan, Liu, Pan, Zhang, Zhang and Wang.)

Macleaya cordata (Willd.) R. Br. is a perennial herbaceous medicinal plant (Papaveraceae) commercially cultivated in China which has been studied for detumescence, detoxification, and insecticidal effect (Lin et al. 2018). In August 2021, anthracnose was observed in 2-year-old M. cordata plants in Benxi county, northeast China (41°45'48″N, 123°69'15″E). Dozens of irregular reddish-brown spots (3-11 mm) were observed on each diseased leaf. The lesions were covered with a layer of gray-white mycelia. As the disease progressed, the spots became necrosis and perforation or they would merged into large lesions, ultimately resulting in wilted leaves (Fig. 1). More than 33% of the plants in a 16-ha field were infected in 2021. The diseased leaves were collected and cut into 3-8 mm pieces, surface-disinfested by immersing them into 1% NaOCl for 2 min, and rinsed three times with sterile distilled water. They were then dried with sterilized absorbent paper, placed on PDA medium amended with chloramphenicol (40 mg/L), and incubated in darkness at 25°C with a 12-h photoperiod. Twenty isolates (BLH1 to 20) were obtained and purified using a single-spore method. Isolate BLH12 was identified and used for the pathogenicity test. Colonies were sparsely fluffy with smooth edges, and gradually became gray to pale orange from the initial white. The underside of the colonies was pale orange towards the center. Conidia were single-celled, cylindrical, and transparent with broadly blunt ends, measuring (15.13 ± 1.14) × (5.80 ± 0.60) μm (n=50). Appressoria were single-celled, brown-to-dark brown, usually elliptical or irregular, and sometimes lobed. Setae were not observed. The isolate was initially identified as Colletotrichum gloeosporioides complex (Prihastuti et al. 2009). The identification was confirmed as described previously (Weir et al. 2012). The rDNA internal transcribed spacer region (OP415560), the glyceraldehyde-3-phosphate dehydrogenase (OP433642), chitin synthase (OP433643), calmodulin (OP433644), actin (OP433645), glutamine synthetase (OP433646), β-tubulin (OP433647), and superoxide dismutase (OP433648) gene sequences were obtained (Carbone & Kohn 1999; Weir et al. 2012), and BLAST searches revealed 99-100% homology with the type culture ICMP 18608 (JX010244, JX010044, JX009683, JX009443, JX009744, JX010078, JX010389, and JX010311). A phylogenetic analysis of combining all loci indicated BLH12 and the type strain of C. aenigma were clustered in one group (Fig. 2). Based on the basis of morphological characteristics and phylogenetic relationships, BLH12 was identified as C. aenigma . For the pathogenicity test, healthy 2-year-old plants were sprayed with a BLH12 spore suspension (1 × 10 5 /mL). Control plants were sprayed with sterile water.There were three replicates (five plants each) per treatment. All plants were incubated at 25°C (12-h photoperiod and 86% relative humidity) and examined after 7 days. The experiment was repeated twice. The inoculated plants showed lesions on the leaf surface, similar to those in the field, whereas the control plants were asymptomatic. The pathogen was successfully reisolated and identified as the methods mentioned above. This fungus reportedly infects the leaves of many woody plants in China (Wang et al. 2020; Zhang et al. 2021). This is the first report of C. aenigma causing anthracnose on M. cordata , which will provide an guideline for developing effective field control practices for the disease.

Ligularia fischeri (Ledeb.) is a perennial herbal plant of Compositae that is cultivated commercially in China as a medicinal, ornamental, and edible plant. Leaf spots were observed in 2-year-old L. fischeri in Benxi County of northeast China, in August 2021. Irregular reddish brown spots ranging from 3 to 11 mm were observed on infected leaves, and each leaf had dozens of spots (Fig. 1). As the disease progressed, the diseased spots withered and the centers fell out, and multiple lesions merge into large diseased spots, causing leaf wilting. The roots and stem bases were not infected during the reproductive stage. More than 37% of the plants in a 18 ha field were infected in 2021. The ten diseased leaves were collected and cut into small (3-5 mm) pieces, which were surface-disinfested by immersing into 1% NaOCl for 2 min and rinsing with sterile distilled water three times. The leaf pieces were then placed on acidified potato dextrose agar (PDA) in petri plates and incubated in the dark at 25°C. Twenty isolates with the same morphological characteristics were obtained. Isolates were further purified by starting a new colony for each isolate from a single spore collected from water agar. Isolate TYTW7 was randomly selected for identification and pathogenicity testing. It grew rapidly and produced profuse aerial mycelia with a carmine red underside. The conidiophores had many fertile branches and formed an elongated stipe with a sphaeropedunculate vesicle at the tip. The one-septate conidia were cylindrical and almost straight with parallel walls and rounded ends. Their sizes ranged from 30.35 to 51.76 × 2.93 to 5.01 μm (n = 100) and the pathogens were initially identified as Calonectria sp. (Crous 2002; Crous et al. 2004; Lombard et al. 2015, 2016). Further confirmation of the identification was determined according to published method (Liu and Chen 2017; Shao and Li 2021). The partial gene regions including the translation elongation factor 1-alpha (GenBank accession no. OP290551), histone H3 (OP290552), calmodulin (OP290553) and β-tubulin (OP290554) were obtained, and BLAST searches showed 99-100% homology with the ex-type culture CERC 8952 (MF527049, MF527065, MF527081 and MF527107) and phylogenetic analysis combining all loci revealed that the isolate TYTW7 and the type strain of Ca. montana clustered in one group (Fig. 2). Based on morphological characteristics and phylogenetic analysis, isolate TYTW7 was identified as Ca. Montana . Healthy 2-year-old plants were used for the pathogenicity test. A spore suspension (1×10 5 spores/mL water) was used to inoculate three host plants; sterile water was sprayed on the same number plants serving as a control. The experiment was repeated three times. All plants were incubated at 27±2°C (12h photoperiod) and were evaluated after seven days. The inoculated plants showed lesions on the leaf surface, similar to those in the field, and the control remained symptomless. The pathogens were successfully reisolated and identified by sequencing, and no pathogens were isolated from symptomless control plants. To our knowledge, this is the first report of Ca. montana causing L. fischeri leaf spot. The disease poses a threat to the production and more control strategies are needed on management options to minimize losses.

Brachybotrys paridiformis Maxim. ex Oliv. (Boraginaceae) is a perennial medicinal plant and vegetable that is cultivated commercially in China. Anthracnose is a devastating disease of B. paridiformis , with annual production losses exceeding 33% based on our survey. In July 2021, anthracnose of B. paridiformis was observed on 2-year-old plants in Shenyang city, Northeast China, which is the most important region for B. paridiformis cultivation. Round or irregular-shaped black spots were exhibited on leaves, with the leaf edges most commonly infected. As the necrosis expanded, the leaves withered and dropped; young leaves were generally not infected (Fig. 1). More than 40% of the plants in a 21-ha sampling field were infected in 2021. Symptomatic leaves (n = 20) were collected and the diseased tissue was cut into small pieces, immersed in 1% NaOCl for 2 min, rinsed three times with sterile water, and placed on acidified potato dextrose agar (PDA) in Petri dishes. After a 3-day incubation in darkness at 25 °C, 18 suspected single-pure morphologically identical Colletotrichum isolates were obtained and sequenced. Isolate SQZ9 was randomly selected and identified. Colonies on PDA were initially white, but gradually became pale brownish with a reverse side that was pale yellowish to pinkish. Aerial mycelia were grayish-white, dense, and cottony, with microsclerotia detected on some aging mycelia. The detected single-celled conidia (11.65-17.25 × 4.25-6.15 µm; n = 50) were fusiform to cylindrical with obtuse to slightly rounded ends. Appressoria were ovoid to clavate and medium brown. Setae were not observed. The morphological characteristics were similar to those of Colletotrichum spp. (Prihastuti et al. 2009; Weir et al. 2012). Initial BLAST searches of the GenBank database revealed the SQZ9 rDNA internal transcribed spacer region (OP389109, 566 bp), glyceraldehyde-3-phosphate dehydrogenase (OP407730, 260 bp), chitin synthase (OP407731, 301 bp), calmodulin (OP407732, 712 bp), actin (OP407733, 282 bp), glutamine synthetase (OP407734, 909 bp), β-tublin (OP407735, 498 bp), and superoxide dismutase (OP407736, 396 bp) sequences were respectively 99%-100% similar to the C. siamense type strain JX010278, JX010019, JX009709, GQ856775, GQ856730, JX010100, JX010410, and JX010332 sequences (Carbone & Kohn 1999; Moriwaki & Tsukiboshi 2009; Stephenson et al. 1997). The SQZ9 identity was confirmed by constructing a phylogenetic tree combining all loci, which grouped the isolate and the C. siamense type strain in the same clade (Fig. 2). For pathogenicity tests, 15 healthy 2-year-old plants (3 plants per pot) were spray-inoculated with SQZ9 conidial suspension (1 × 105 conidia/mL) at 2 mL per plant. Same number of plants sprayed with water were used as control. This experiment was repeated twice. All plants were covered with clear plastic bags for 72 h to maintain high humidity and then placed in a greenhouse (29 °C, natural light, and 85% relative humidity). After six days, the inoculated leaves exhibited symptoms that were similar to those observed in the field, but the controls were symptomless. The same fungus was recovered from inoculated symptomatic leaves, and its identity was confirmed by sequencing and a phylogenetic analysis. This is the first report of C. siamense causing anthracnose on B. paridiformis in China. Future studies should assess the effectiveness of chemical and biological control measures for managing this disease.

Background: Cancer is considered one of the most lethal diseases worldwide. Venous thromboembolism (VTE) is the second leading cause of death in cancer patients. As one of the most reproducible predictors of thromboembolism, the D-dimer level is commonly considered by oncologists. Previous studies have demonstrated that the most correlated genes at the D-dimer level are F3, F5 and FGA. Methods: Using data from TCGA and multiple webtools, including GEPIA2, UALCAN, TIMER2.0, Kaplan-Meier Plotter and CIBERSORTx, we analyzed the tumor mutation burden (TMB), microsatellite instability (MSI) and functions of D-dimer-related genes in cancer. Validation was conducted via quantitative real-time polymerase chain reaction (qRT-PCR) and independent GEO + GTEx cohort. All statistical analyses were performed in R software and GraphPad Prism 9. Results: F3, F5 and FGA were expressed differently in multiple cancer types. TMB, MSI and anti-PD1/PDL1 therapy responses were correlated with D-dimer-related gene expression. D-Dimer-related genes expression affect the survival of cancer patients. F3 and F5 functioned in TGF-beta signaling. F3 and F5 were related to immunity and affected the fraction of CD8 + T cells by upregulating the TGF-beta signaling pathway, forming an F3, F5/TGF-beta signaling/CD8+ T cell axis. Conclusion: F3, F5 and FGA serve as satisfactory GC multibiomarkers and potentially influence the immune microenvironment and survival of cancer patients by influencing TGF-beta signaling., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Guan, Xu, Sui, Chen, Luan, Jiang, Wei, Long, Zhao, Han, Xu, Lin and Guan.)

In the past 3 decades, the cartilage repair potential of mesenchymal stromal cells, or mesenchymal stem cells (MSCs), has been widely examined in animal studies. Unfortunately, the phenotype and physical properties of MSC-derived cartilage tissue are not comparable to native hyaline cartilage. In particular, chondrocytic hypertrophy, a phenotype that is not observed in healthy hyaline cartilage, is concomitant with MSC chondrogenesis. Given that hypertrophic chondrocytes potentially undergo apoptosis or convert into osteoblasts, this undesired phenotype needs to be prevented or minimized before MSCs can be used to repair cartilage injuries in the clinic. In this review, we first provide an overview of chondrocytic hypertrophy and briefly summarize current methods for suppressing hypertrophy in MSC-derived cartilage. We then highlight recent progress on modulating the canonical Wnt/β-catenin pathway for inhibiting hypertrophy. Specially, we discuss the potential crosstalk between Wnt/β-catenin with other pathways in regulating hypertrophy. Lastly, we explore future perspectives to further understand the role of Wnt/β-catenin in chondrocytic hypertrophy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wang, Guan, Xiang, Clark, Alexander, Simonian, Deng and Lin.)

The tumor microenvironment (TME) comprises distinct cell types, including stromal types such as fibroblast cells and macrophage cells, which have recently become a critical factor in tumor development and progression. Here, we identified the TME-related gene, plexin domain containing 2 (PLXDC2), in a high-stromal-score population. And we revealed that this gene was related to poor survival and advanced (tumor-node-metastasis) stage in gastric cancer (GC) patients from The Cancer Genome Atlas database. An integrated gene profile and functional analysis of the proportions of tumor-infiltrating immune cells revealed that the expression of the M2 macrophages cell marker CD163 was positively correlated with PLXDC2 expression. In addition, the M2 macrophages gene signature and high PLXDC2 expression were associated with the inflammatory signaling pathway and the epithelial-to-mesenchymal transition (EMT)-related gene signature. Single-cell study of GC identified PLXDC2 was enriched specifically in fibroblasts and monocytes/macrophages populations, which supported its important role in the stroma. Furthermore, according to a tissue microarray immunohistochemistry analysis, the expression of PLXDC2 elevated in human GC stromal specimens compared to tumor tissue specimens. Moreover, PLXDC2 overexpression in the stromal compartment was associated with CD163-positive regulatory M2 macrophages, and its functions were related to the pathogenesis of GC. Multiplexed immunohistochemistry verified PLXDC2's correlation with EMT markers. Our data suggested that PLXDC2 was expressed in stromal cells and that its crosstalk with tumor-associated macrophages could contribute to cancer biology by inducing the EMT process., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Guan, Du, Wang, Gou, Xue, Xu, Li, Chan, Wu, Xu, Ni, Xu and Hu.)