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Abstract Studying the carbon effect of land use in watersheds is important for mitigating global warming, promoting coordinated emission reduction in different regions within the watersheds, and realizing high-quality development of the watersheds. Although a number of scholars have carried out relevant studies in the past, they mainly focused on carbon emissions, rarely involved the carbon balance formed by carbon sources and sinks, and lacked relevant studies on the development of low-carbon economy sub-region. Based on this, this study takes the Yellow River Basin as an example, explores the spatial and temporal evolution of carbon emissions from land use in counties in the Yellow River Basin from 1980 to 2020, and predicts the spatial pattern of carbon income and expenditure from land use under natural conditions in 2030 and 2060 using the PLUS model; and then superimposes on the main functional area planning, divides 735 counties in the Yellow River Basin into six low-carbon economic development subregions, and analyzes their economic development The model of their economic development is analyzed. The results show that: (1) the spatial and temporal differentiation of land use carbon balance in the Yellow River Basin has changed greatly over the past 40 years, (2) the spatial distribution pattern of land use carbon balance in the natural context in 2030 and 2060 is more similar to that in 1990, (3) the carbon emission reduction potentials and pattern optimization of the different low-carbon economic development subregions differ greatly, and they have different low-carbon economic development patterns. The results of this study provide a theoretical basis for scientifically and rationally formulating economic policies for low-carbon development in the counties of the Yellow River Basin, and also provide an important reference for related studies in other similar basins or regions in the world.

Panax notoginseng is a traditional Chinese medicine rich in many pharmacological components. The root of the ‘Miaoxiang Sanqi No. 2’ is yellow or greenish yellow, while a novel cultivar-‘Wenyuan Ziqi No. 1’ shows purple root and is thought to have high medicinal value. Little information is available about the anthocyanin biosynthesis in P. notoginseng root. In this study, we compared the ‘Miaoxiang Sanqi No. 2’ and ‘Wenyuan Ziqi No. 1’ in morphological, transcriptional and metabolic levels. The results showed that purple rich in the periderm, rhizome and phloem around cambium of the ‘Wenyuan Ziqi No. 1’ root and cyanidin 3-O-galactoside was the main anthocyanin causing purple. Moreover, ‘Wenyuan Ziqi No. 1’ highly accumulated in 155 metabolites, including flavones, phenylpropanoids and lipids. Transcriptome data showed that phenylpropanoid biosynthesis pathway genes are highly expressed in ‘Wenyuan Ziqi No. 1’. Conjoint analysis showed that anthocyanin biosynthesis pathway substances were highly accumulated in ‘Wenyuan Ziqi No. 1’, and the expression level of structural genes involved in anthocyanin biosynthesis pathway was higher in ‘Wenyuan Ziqi No. 1’. Meanwhile, eight R2R3-MYB genes that might be involved in anthocyanin biosynthesis were identified. The comprehensive analysis of two cultivars provides new insights into the understanding of root coloration in P. notoginseng.

A sensitive and specific bioanalytical method was required to measure the exposure of a LAGA-mutated surrogate mouse IgG2a monoclonal antibody in mouse plasma, but the lack of highly specific reagents for the LAGA mutant hindered the development of a ligand-binding assay. Equally problematic is that no sensitive unique tryptic peptides suitable for quantitative mass spectrometric analysis could be identified in the mIgG2a complementarity-determining regions. To overcome these challenges, a trypsin alternative pepsin, an aspartic protease, was systematically investigated for its use in digesting the mutated mIgG2a antibody to allow generation of signature peptides for the bioanalytical quantification purpose. After a series of evaluations, a rapid one-hour pepsin digestion protocol was established for the mutated Fc backbone. Consequently, a new pepsin digestion-based liquid chromatography-tandem mass spectrometry (LC/MS/MS) method was successfully developed to support the mouse pharmacokinetic (PK) sample analysis. In brief, robust and reproducible C-terminal cleavage of both leucine and phenylalanine near the double mutation site of the mutated mIgG2a was accomplished at pH ≤2 and 37°C. Combined with a commercially available rat anti-mIgG2a heavy-chain antibody, the established immunoaffinity LC/MS/MS assay achieved a limit of quantitation of 20 ng/mL in the dynamic range of interest with satisfactory assay precision and accuracy. The successful implementation of this novel approach in discovery PK studies eliminates the need for tedious and costly generation of specific immunocapturing reagents for the LAGA mutants. The approach should be widely applicable for developing popular LAGA mutant-based biological therapeutics.

The production of huge amounts of biogas slurry during livestock breeding has resulted in pressing environmental issues. Although paddy fields can be potential sinks for the disposal of biogas slurry, the impacts of biogas slurry on rice production, grain quality, and relevant environmental risks in the Yangtze Delta region remain unclear. Herein, we conducted a field trial from 2021 to 2023 which involved different gradients of biogas slurry utilization, including CK (no fertilizer), CN (100% chemical nitrogen (N) of 240 kg ha−1), NBS (biogas slurry replacing 50% chemical N), BS1 (replacing 100% chemical N), BS1.5 (replacing 150% chemical N), and BS2 (replacing 200% chemical N). The results showed that there were no significant differences in average rice yields between CN, NBS, BS1.5, and BS2 over the three-year study period, with an average yield of 8283 kg ha−1, and the average yields of BS1 and CK were 7815 kg ha−1 and 6236 kg ha−1, respectively. However, heavy utilization of biogas slurry (BS1.5 and BS2) not only significantly reduced the rice seed-setting rate, the 1000-grain weight, and the processing quality, but also significantly increased the protein, amylose, Cu, and Zn content in rice grains; additionally, higher N losses occurred via surface water and increased NH3 volatilization was observed, finally resulting in lower nitrogen-use efficiency. Meanwhile, moderate utilization of biogas slurry (NBS and BS1) led to better rice quality and nitrogen-use efficiency, lower potential food safety risk, and N loss. Further, compared to BS1, NBS showed higher yield, harvest index, processing quality, gel consistency, palatability scores, and nitrogen-use efficiency, but lower N losses were present. Overall, the NBS treatment balanced the agronomic benefits and environmental risks in the Yangtze River Delta region. In the future, more attention should be paid to food safety and environmental risks when using biogas slurry.

As the main contributor to greenhouse gas (GHG) in paddy soil, information on methane (CH4) emission characteristics under different tillage and cultivation practices are limited. A five-year field trial was conducted from 2019 in a single-cropping rice system in Taihu Lake region, east of China. The experiment had a completely randomized block design, and the treatments included rotary tillage plus rice dry direct seeding (RD), rotary tillage plus rice mechanical transplanting (RT), and plowing tillage plus rice mechanical transplanting (PT). We determined the rice yield, GHG emission, soil traits, and methanogens and methanotrophs in 2022 and 2023. The results revealed that PT and RT significantly increased rice yield compared to RD, whereas PT simultaneously increased CH4 emissions. The year-averaged cumulative CH4 emissions in PT were increased by 38.5% and 61.4% higher than RT and RD, respectively. Meanwhile, yield-scaled global warming potentials (GWPs) in RT and RD were lower than those in PT. Tillage and cultivation practices shifted mcrA and pmoA abundances, and PT significantly decreased pmoA abundance. The community structure and diversity of the methanogens and methanotrophs were not significantly affected. Structural equation model analyses illustrated that CH4 emissions were regulated by mcrA and pmoA directly, which in turn, regulated by soil carbon and nitrogen. Overall, rotary tillage plus mechanism transplanting was a feasible agronomic technology in a single-cropping rice system in Taihu Lake region, exhibiting higher and more stable rice productivity, accompanied with lower CH4 emissions and yield-scaled GWP.

Abstract Background The phyllosphere mycobiome plays a crucial role in plant fitness and ecosystem functions. The complex microbial ecological networks (MEN) formed by these fungi remain poorly understood, particularly with regard to their organization strategy and their contributions to plant secondary metabolites such as saponin. Results In this study, we constructed six MENs from leaf epiphytic and endophytic mycobiomes of three Panax species distributed in the northeast and southwest ends of mainland China. Hub nodes were absent in these MENs, which were significantly more complex, robust, and less efficient compared to random graphs (P 0.5). Taxonomic analysis revealed that Pleosporales and Chaetothyriales were significantly overrepresented by positive regulation taxa (PRT) of total saponin content (FDR

Abstract Background Histone acetyltransferases (HATs) play an important role in plant growth and development, stress response, and regulation of secondary metabolite biosynthesis. Hemp (Cannabis sativa L.) is famous for its high industrial, nutritional, and medicinal value. It contains non-psychoactive cannabinoid cannabidiol (CBD) and cannabinol (CBG), which play important roles as anti-inflammatory and anti-anxiety. At present, the involvement of HATs in the regulation of cannabinoid CBD and CBG synthesis has not been clarified. Methods The members of HAT genes family in hemp were systematically analyzed by bioinformatics analysis. In addition, the expression level of HATs and the level of histone acetylation modification were analyzed based on transcriptome data and protein modification data. Real-time quantitative PCR was used to verify the changes in gene expression levels after inhibitor treatment. The changes of CBD and CBG contents after inhibitor treatment were verified by HPLC-MS analysis. Results Here, 11 HAT genes were identified in the hemp genome. Phylogenetic analysis showed that hemp HAT family genes can be divided into six groups. Cannabinoid synthesis genes exhibited spatiotemporal specificity, and histones were acetylated in different inflorescence developmental stages. The expression of cannabinoid synthesis genes was inhibited and the content of CBD and CBG declined by 10% to 55% in the samples treated by HAT inhibitor (PU139). Results indicated that CsHAT genes may regulate cannabinoid synthesis through altering histone acetylation. Conclusions Our study provides genetic information of HATs responsible for cannabinoid synthesis, and offers a new approach for increasing the content of cannabinoid in hemp.

A human induced pluripotent stem cell (iPSC) line (SDASi001-A) was generated from patient with Schimke immune-osseous dysplasia (SIOD), carrying heterozygous mutations in SMARCAL1 gene. Peripheral blood mononuclear cells (PBMCs) were reprogrammed using non-integrating delivery of OCT4, SOX2, KFL4, BCL-XL and c-MYC. The iPSC line expresses pluripotency markers, displays a normal karyotype, and has the ability to differentiate into cells of three germ layers in vitro. This iPSC line represents a valuable cell model for SIOD in humans.

Medicinal and edible seed Semen Persicae is susceptible to mycotoxin and fungal contamination. However, the occurrence of mycotoxin contamination and fungal infection is still unclear. In this paper, ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry and high-throughput sequencing were conducted to determine the mycotoxin contents and fungal abundances of Semen Persicae. 42.86% of samples were positive for aflatoxin B1 (26.48–48.37 μg/kg) and 28.57% of samples were positive for aflatoxin B2 (1.47–4.82 μg/kg). Ochratoxin A and fumonisin B1 were only detected in one sample (91.02 and 34.61 μg/kg, respectively). Chao 1 and Shannon indices were significantly higher in the Dalian of Liaoning, Baotou of Innermongolia and Langfang of Hebei regions than in other groups. Ascomycota, Basidiomycota, Wallemia, Candica, Saccharomyces and Aspergillus were the predominant fungi and they were significantly region-specific. Simultaneously, the diversity, composition and co-occurrence network complexity in the mycotoxin-free group were significantly higher than those in the mycotoxin-contaminated group. Spearman correlation analysis showed aflatoxins, ochratoxin A and fumonisins contents were positively and significantly correlated with the abundances of Aspergillus, Rhodotorula, Wallemia and Candida. In conclusion, this study reported the prevalence of mycotoxin contamination and the great diversity of fungi associated with Semen Persicae for the first time, providing an early warning for subsequent potential mycotoxin biosynthesis.

IntroductionArecae semen, which is derived from the dried ripe seed of Areca catechu L., has been commonly used as one of the major traditional Chinese medicines (TCMs). Three types of crude herbal preparations, namely, raw Arecae semen (AS), Arecae semen tostum (SAS), and Arecae semen carbonisata (FAS), are available for different clinical applications in TCMs. Although aflatoxin contamination in Arecae semen has been reported preliminarily, only a few studies have been conducted on fungal contamination.MethodsIn this study, the presence of fungi on the surface of three Arecae semen (AS, SAS, and FAS) that collected from four provinces were investigated using high-throughput sequencing and internal transcribed spacer 2.ResultsResults showed that the phyla Ascomycota (75.45%) and Basidiomycota (14.29%) and the genera Wallemia (7.56%), Botryosphaeria (6.91%), Davidiella (5.14%), and Symbiotaphrina (4.87%) were the dominant fungi, and they presented significant differences in four areas and three processed products (p

BackgroundRenal involvement is rarely reported in juvenile dermatomyositis and may be caused by the toxic effects of myoglobinuria or an autoimmune reaction. We report a case of dermatomyositis and nephrotic syndrome in a child to explore the association between juvenile dermatomyositis and renal involvement.Case presentationAn 8-year-old girl with skin rash, edema, proximal muscle weakness predominantly involving the lower extremities, low-grade fever, and foamy urine was admitted to our hospital. Her laboratory tests met the criteria of nephrotic syndrome. She had elevated creatine kinase and lactate dehydrogenase and was diagnosed with juvenile dermatomyositis after electromyography and muscle MRI. Anti-NXP2 antibodies were positive. Her proteinuria was relieved soon after treatment with prednisone and methotrexate, but her muscle strength progressively decreased. The disease was relieved after pulse methylprednisolone treatment and mycophenolate mofetil, but recurred after drug reduction with mild proteinuria. Adalimumab was used for treatment and helped reduce the doses of glucocorticoid and mycophenolate mofetil.ConclusionJuvenile dermatomyositis may be one of the rare causes of nephrotic syndrome. The mechanism involved in JDM combined with renal injury may be multifactorial. Autoantibodies may play important roles in both muscle and renal damage.

Abstract Background Codonopsis pilosula, an important medicinal plant, can accumulate certain metabolites under moderate drought stress. Endophytes are involved in the metabolite accumulations within medicinal plants. It is still unknown that the endophytes of C. pilosula are associated with the accumulations of metabolites. This study aims to investigate the promoting effect of endophytes on the accumulations of active substances in C. pilosula under drought stress. Methods High–performance liquid chromatography and high–throughput sequencing technology were performed to investigate changes in the contents of secondary metabolite and endophyte abundances of C. pilosula under drought stress, respectively. Spearman’s correlation analysis was further conducted to identify the endophytic biomarkers related to accumulations of pharmacodynamic compounds. Culture-dependent experiments were performed to confirm the functions of endophytes in metabolite accumulations. Results The distribution of pharmacological components and diversity and composition of endophytes showed tissue specificity within C. pilosula. The contents of lobetyolin, syringin, and atractylolide III in C. pilosula under drought stress were increased by 8.47%‒86.47%, 28.78%‒230.98%, and 32.17%‒177.86%, respectively, in comparison with those in untreated groups. The Chao 1 and Shannon indices in different parts of drought–stressed C. pilosula increased compared with those in untreated parts. The composition of endophytic communities in drought treatment parts of C. pilosula was different from that in control parts. A total of 226 microbial taxa were identified as potential biomarkers, of which the abundances of 42 taxa were significantly and positively correlated to the pharmacodynamic contents. Culture-dependent experiments confirmed that the contents of lobetyolin and atractylolide III were increased by the application of Epicoccum thailandicum, Filobasidium magnum, and Paraphoma rhaphiolepidis at the rates of 11.12%‒46.02%, and that the content of syringin was increased by Pseudomonas nitroreducens at the rates of 118.61%‒119.36%. Conclusions Certain endophytes participated in the accumulations of bioactive metabolites, which provided a scientific evidence for the development and application of microorganisms to improve the quality of traditional Chinese medicine.

Estuarine tidal flats are located in the land-sea crisscross-sensitive zone. The degraded and shrinking state of major tidal flat landscapes weakens their ecological role in maintaining the safety of coastal wetlands in the world. However, little attention has been paid to tidal flat landscape evolution and ecological risks due to the tidal flats being located in the periphery of estuarine wetlands. This study was conducted in the tidal flats of the Liaohe estuary wetlands. The GIS spatial analysis and a transition matrix were combined to explore the tidal flat landscape evolution characteristics. The ecological risk faced by the tidal flats was evaluated by a landscape ecological risk model constructed by landscape indexes. The key driving factors and threshold values that affect the ecological risks were clarified using geographical detectors and regression analysis methods. The results showed that the tidal flats mainly evolved into a natural wetland before 2000 and rapidly changed into an artificial landscape between 2000 and 2010; the proportion of artificial landscapes then began to decline after 2010. Over the past 30 years, the landscape ecological risk in this area has generally been at a medium risk level. The ecological risk index showed a decreasing trend with the increases in landscape diversity. The low-risk artificial plaques have gradually replaced high-risk and medium-risk plaques of exposed tidal flats and natural wetlands. The study showed that human development activities were the key factors affecting the landscape ecological risks of the tidal flats. River sediment discharge was the key natural factor in maintaining the landscape ecological risk at a stable level. Therefore, the human development activities should be controlled within an appropriate limit, and the material supply of the upstream basin should be guaranteed to maintain the stability of the tidal flat landscape form in the protection and utilization of estuarine tidal flats in the future.

Revealing the ecological patterns of abundant and rare microbial subcommunities in rhizosphere of cultivated plant is critical for understanding plant–microbe interactions and the formation of functionally important rhizosphere microbiome. Based on rhizodeposit support effects and colonization priority effects, the present study outlined two disparate hypotheses predicting the more deterministic assembly of abundant (the “dominance-determinism” hypothesis) and rare subcommunities (the “rare-determinism” hypothesis), respectively. These hypotheses were tested using bacterial and fungal subcommunities in the rhizosphere of three cultivated Panax species (P. ginseng, P. quinquefolium, and P. notoginseng). In each plant species, the abundant and rare taxa exhibited different patterns of distribution and composition, but similar edaphic factors could explain their phylogenetic community variations. Network analysis indicated that the abundant taxa located in more central positions than the rare taxa, and the negative links mainly exhibited between abundant and rare taxa. Null model showed that stochastic and deterministic processes dominated the assembly of abundant (determinism: 11.97–45.30 %) and rare (determinism: 67.24–96.30 %) subcommunities, respectively. In addition, almost all members of the rhizosphere abundant taxa (> 89 %) were also highly abundant in corresponding bulk soil. Among the three Panax species, the rare taxa exhibited stronger taxonomic overlap and phylogenetic similarity compared to the abundant taxa. These results indicate that the assembly of abundant subcommunities in Panax rhizosphere is highly stochastic and may relate to the priority effects, whereas the rare taxa tend to be the results of plant selection and exhibit potentially evolutionary conservatism.

BackgroundWith the development trend of healthy aging and intelligent integration, escort products have become a new means of healthy aging. Healthy old-age care pays attention to the convenience and informatization of life. To meet the needs, designers often design multiple accompanying product solutions, and it is very important to use reasonable evaluation methods to decide on the optimal solution.PurposesA new comprehensive evaluation method is proposed to reduce the subjectivity and one-sidedness of the selection process of intelligent escort product design solutions, and to make the decision more objective and reasonable. Such decisions can enhance the experience and naturalness of the elderly using intelligent products.MethodsFirst, a large number of user interviews were analyzed using the grounded theory, gradually refine through theoretical coding, and abstracted with the design scheme evaluation index. Second, the idea of game-theoretic weighting is used to optimize a linear combination of subjective and objective weights to determine the final weights of each evaluation indicator. Finally, the evaluation and selection are completed based on the solution ranking determined by the approximate ideal solution ranking method (TOPSIS). It is applied for the selection of the elderly escort robot design, and the usability test is conducted using the PSSUQ to verify the selection results.ResultsA new comprehensive evaluation method can better complete the preferential selection of product design solutions for healthy aging escorts, and reduce the subjectivity and one-sidedness of the evaluation.ConclusionThis method compensates for the reliance on personal experience in the selection of options, and improve the subjectivity of the evaluation index determination process and the deviation of index weighting. Improving the objectivity and scientificity of decision-making reduces the blindness of design and production. It also provides a theoretical reference for the research scholars of healthy aging companion products.

Shaogan Fuzi Decoction (SGFD), one of the classical prescriptions of Chinese Medicine, has a long history in the treatment of rheumatoid arthritis (RA), but definitive studies on its efficacy and mechanism of action are lacking. This study aims to elucidate the pharmacodynamic role of SGFD against RA and the potential mechanisms based on a combination of network pharmacology and experimental verification. The RA model in rats was induced by intradermal injection of bovine type Ⅱ collagen and incomplete Freund’s adjuvant at the tail root. SGFD was administered once a day by oral gavage for 4 weeks. After SGFD administration, rat’s arthritis index (AI) score and paw swelling decreased to some extent, and synovial inflammation, vascular hyperplasia, and cartilage destruction of the ankle joint were improved. Simultaneously, thymus and spleen index and serum levels of C-reactive protein (CRP) were lowered. Network pharmacology revealed that quercetin, kaempferol, naringenin, formononetin isorhamnetin and licochalcone A were the potentialiy active components, and IL6, TP53, TNF, PTGS2, MAPK3 and IL-1β were potential key targets for SGFD in the treatment of RA. Ingredients-targets molecular docking showed that the components had the high binding activity to these target proteins. The mechanism of SGFD for RA involves various biological functions and is closely correlated with TNF signaling pathway, Osteoclast differentiation, T cell receptor signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, NF-κB signaling pathway, toll-like receptor signaling pathway, and so on. Western blot and ELISA showed that the expression of toll-like receptor 4 (TLR4), nuclear factor kappa-B (NF-κB) p65, phosphorylated c-Jun N-terminal kinase (p-JNK), p-p38, phosphorylated extracellular regulated kinase (p-ERK) and TNF-α was significantly upregulated in the synovium of RA rats, and the levels of serum inflammatory factors were significantly increased. SGFD inhibits the activation of the TLR4/NF-κB/MAPK pathway and the expression/production of pro-inflammatory cytokines. In summary, SGFD could improve the symptoms and inflammatory response in collagen-induced arthritis (CIA) rat model. The mechanism might be related to the regulation of TLR4/MAPKs/NF-κB signaling pathway and the reduction of inflammatory factor release, which partially confirms the results predicted by network pharmacology.

Summary Endophytes may participate in the conversion of metabolites within medicinal plants, influencing the efficacy of host. However, the distribution of endophytes within medicinal plants P. notoginseng and how it contributes to the conversion of saponins are not well understood. Here, we determined the distribution of saponins and endophytes within P. notoginseng compartments and further confirm the saponin conversion by endophytes. We found metabolites showed compartment specificity within P. notoginseng. Potential saponin biomarkers, such as Rb1, Rg1, Re, Rc and Rd, were obtained. Endophytic diversity, composition and co‐occurrence networks also showed compartment specificity, and bacterial alpha diversity values were highest in root compartment, consistently decreased in the stem and leaf compartments, whereas those of fungi showed the opposite trend. Potential bacterial biomarkers, such as Rhizobium, Bacillus, Pseudomonas, Enterobacter, Klebsiella, Pantoea and fungal biomarkers Phoma, Epicoccum, Xylariales, were also obtained. Endophytes related to saponin contents were found by Spearman correlation analysis, and further verification experiments showed that Enterobacter chengduensis could convert ginsenoside Rg1 to F1 at a rate of 13.24%; Trichoderma koningii could convert ginsenoside Rb1 to Rd at a rate of 40.00% and to Rg3 at a rate of 32.31%; Penicillium chermesinum could convert ginsenoside Rb1 to Rd at a rate of 74.24%.

Hemp seeds are rich in metabolites such as protein, lipids and flavonoids, which are beneficial to health and can be used as a nutritional supplement. Few studies have focused on the metabolites of different hemp seed varieties. In the current study, using widely targeted metabolomics based on UHPLC-QQQ-MS/MS, we compared the metabolomes of seeds from seven hemp varieties with different uses. A total of 1,001 metabolites, including 201 flavonoids, 86 alkaloids, and 149 phenolic acids, were identified. Flavonoids, organic acids, alkaloids, lipids, and fatty acids with high nutritional value are important to investigate the differences between hemp accessions. By using weighted gene co-expression network analysis (WGCNA), six modules of closely related metabolites were identified. And, we identified the metabolite characteristics and hub metabolites of each variety. Then, we experimentally determined antioxidant activity of seven varieties and demonstrated that alkaloids, flavonoids, phenolic acids, terpenes, and free fatty acids are responsible for the antioxidant activity of hemp seeds. Our research provides useful information for further investigation of the chemical composition of hemp seeds.

Abstract Sophora flavescens are widely used for their pharmacological effects. As its main pharmacological components, alkaloids and flavonoids are distributed in the root tissues wherein molecular mechanisms remain elusive. In this study, metabolite profiles are analyzed using metabolomes to obtain biomarkers detected in different root tissues. These biomarkers include alkaloids, phenylpropanoids, and flavonoids. The high-performance liquid chromatography analysis results indicate the differences in principal component contents. Oxymatrine, sophoridine, and matrine contents are the highest in the phloem, whereas trifolirhizin, maackiain, and kushenol I contents are the highest in the xylem. The transcript expression profiles also show tissue specificity in the roots. A total of 52 and 39 transcripts involved in alkaloid and flavonoid syntheses are found, respectively. Among them, the expression levels of LYSA1, LYSA2, AO2, AO6, PMT1, PMT17, PMT34, and PMT35 transcripts are highly and positively correlated with alkaloids contents. The expression levels of 4CL1, 4CL3, 4CL12, CHI5, CHI7, and CHI9 transcripts are markedly and positively correlated with flavonoids contents. Moreover, the quantitative profiles of alkaloids and flavonoids are provided, and the pivotal genes regulating their distribution in S. flavescens are determined. These results contribute to the existing data for the genetic improvement and target breeding of S. flavescens.

Cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) is a check point protein expressed on the surface of T cells and plays a central role in regulating the immune response. In recent years, CTLA-4 has become a popular target for cancer immunotherapy in which blocking CTLA-4 can restore T-cell function and enhance the immune response against cancer. Currently, there are many CTLA-4 inhibitors in a variety of modalities, including cell therapies, which are being developed in both preclinical and clinical stages to further harness the potential of the target for the treatment of certain types of cancer. In drug discovery research, measuring the level of CTLA-4 in T cells is important for drug discovery and development because it provides key information for quantitative assessment of the pharmacodynamics, efficacy, and safety of the CTLA-4-based therapies. However, to our best knowledge, there is still no report of a sensitive, specific, accurate, and reliable assay for CTLA-4 measurement. In this work, an LC/MS-based method was developed to measure CTLA-4 in human T cells. The assay demonstrated high specificity with an LLOQ of 5 copies of CTLA-4 per cell when using 2.5 million T cells for analysis. As shown in the work, the assay was successfully used to measure CTLA-4 levels in subtype T-cell samples from individual healthy subjects. The assay could be applied in supporting the studies of CTLA-4-based cancer therapies.

Background: Panax quinquefolius and Panax notoginseng are widely used and well known for their pharmacological effects. As main pharmacological components, saponins have different distribution patterns in the root tissues of Panax plants. Methods: In this study, the representative ginsenosides were detected and quantified by desorption electrospray ionization mass spectrometry and high-performance liquid chromatography analysis to demonstrate saponin distribution in the root tissues of P. quinquefolius and P. notoginseng, and saponin metabolite profiles were analyzed by metabolomes to obtain the biomarkers of different root tissues. Finally, the transcriptome analysis was performed to demonstrate the molecular mechanisms of saponin distribution by gene profiles. Results: There was saponin distribution in the root tissues differed between P. quinquefolius and P. notoginseng. Eight-eight and 24 potential biomarkers were detected by metabolome analysis, and a total of 340 and 122 transcripts involved in saponin synthesis that were positively correlated with the saponin contents (R > 0.6, P

Rhizosphere microbiome promotes plant growth; however, the succession of rhizosphere microbial community during the growth stages of perennial medicinal plant Panax notoginseng (P. notoginseng) is still unclear. Here, amplicon sequencing was performed to assess the succession characteristics of rhizosphere microbiomes during developmental stages. Results showed that bacterial and fungal communities were mainly shaped by the development stages. The microbial α-diversities first increased and then decreased with plant growth and the variation in microbial composition was active at the 3-year root growth (3YR) stage. The variation trend of cross-domain co-occurrence network complexity was similar to that of α-diversities. Cross-domain nodes decreased at the 3YR stage and fungal nodes increased at the 3YR stage. This study provided a detailed and systematic survey of rhizosphere microbiomes during the growth stages of P. notoginseng. The findings revealed that the development stages of P. notoginseng drove the temporal dynamics of rhizosphere communities. This study helps in harnessing the power of microbiomes to evaluate herbal medicine growth and provides valuable information to guide the microbial breeding of medical plants.

Abstract Background Panax notoginseng is a highly valuable medicinal plant. Reduced P. notoginseng yield is a common and serious problem that arises in a continuous cropping system. Variation in the composition and function of soil microbial community is considered the primary cause of yield reduction. Methods This study used shotgun metagenomic sequencing approaches to describe the taxonomic and functional features of P. notoginseng rhizosphere microbiome and screen microbial taxa and functional traits related to yields. Results At the family and genus level, a total of 43 families and 45 genera (relative abundance > 0.1%) were obtained, and the correlation with the yield of P. notoginseng was further analyzed. Nitrosomonadaceae, Xanthomonadaceae, Mycobacterium and Arthrobacter that were enriched in soils with higher yields were positively correlated with P. notoginseng yields, thereby suggesting that they might increase yields. Negative correlation coefficients indicated that Xanthobacteraceae, Caulobacteraceae, Oxalobacteraceae, Chitinophagaceae, Sphingomonas, Hyphomicrobium, Variovorax and Phenylobacterium might be detrimental to P. notoginseng growth. A total of 85 functional traits were significantly (P

Panax notoginseng (Panax notoginseng (Burk.) F.H. Chen), a plant of high medicinal value, is severely affected by root rot during cultivation. Here, we generated a reference genome of P. notoginseng, with a contig N50 size of 241.268 kb, and identified 66 disease-resistance genes (R-genes) as candidate genes for breeding disease-resistant varieties. We then investigated the molecular mechanism underlying the responses of resistant and susceptible P. notoginseng genotypes to Fusarium oxysporum infection at six time points by RNA-seq. Functional analysis of the genes differentially expressed between the two genotypes indicated that genes involved in the defense response biological process like hormone transduction and plant-pathogen interaction are continuously and highly expressed in resistant genotype during infection. Moreover, salicylic acid and jasmonic acid levels gradually increased during infection in the resistant genotype. Coexpression analysis showed that PnWRKY22 acts as a hub gene in the defense response of the resistant genotype. Finally, transiently overexpressing PnWRKY22 increased salicylic acid levels in P. notoginseng leaves. Our findings provide a theoretical basis for studying root rot resistance in P. notoginseng.

Interactions between plants and microbes may promote the growth of plants and regulate the production of secondary metabolites. Hemp (Cannabis sativa) is an annual herb and an important commercial crop. However, the assembly and network of hemp-associated microbiomes inhabiting in soil and plant compartments have not been comprehensively understood. This work investigated the assembly and network of bacterial and fungal communities living in soils (bulk and rhizosphere) and plant compartments (root, stem, leaf, and flower) of four hemp ecotypes cultivated in the same habitat. Microbiome assembly was predominantly shaped by compartment niche. Microbial alpha diversity was the highest in soil, continually decreased from root to flower. Core bacterial genera Pseudomonas, Bacillus, Rhizobium, Planococcus, and Sphingomonas were mostly enriched in aerial endosphere niches; Clitopilus, Plectosphaerella, and Mortierella were enriched in belowground endosphere. Microbial network complexity and connectivity decreased from root to flower. According to source tracking analysis, hemp microbiota primarily originated from soil and were subsequently filtered in different plant compartments. This work provides details on hemp-associated microbiome along the soil–plant continuum and a comprehensive understanding of the origin and transmission mode of endophytes in hemp.

Knowledge about the ecological patterns of abundant and rare bacteria in rhizosphere is still lacking. Through a large-scale sampling across the arable area of a medicinal plant Panax notoginseng, we examined the community assembly of abundant and rare rhizobacteria as well as their putative functions and co-occurrence patterns. At local level, the rare taxa exhibited significantly stronger phylogenetic clustering than the abundant taxa. At metacommunity level, both two subcommunities showed significant biogeographic patterns, but they were assembled by different ecological processes. Null model analysis showed that dispersal limitation (84.68%) governed the assembly of abundant subcommunity, whereas homogeneous selection (90.11%) dominated the dynamics of rare subcommunity. Although the putative functions indicated that the rare taxa potentially enriched more functional traits associated with plant-microbe interactions, the overall abundance profiles of putative pathways in two subcommunities exhibited significant collinearity. Additionally, the topological properties of abundant taxa in the co-occurrence network were significantly higher than those of rare taxa. We propose that stochasticity and determinism dominate the assembly of abundant and rare subcommunities, respectively. The abundant taxa may exhibit stronger environmental adaptability, while the rare taxa are more likely the result of rhizosphere effect. These findings also emphasize the importance of integrating phylogeny in ecological researches on plant-associated microbiota.

Understanding the ecological patterns of rhizosphere microbial communities is critical for propelling sustainable agriculture and managing ecosystem functions by exploiting microorganisms. However, this knowledge is still unclear, especially under host-associated large-scale and regarding the comparison between bacteria and fungi. We examined community assembly processes and community characters including environmental thresholds and co-occurrence patterns across the cultivatable area of Panax notoginseng for bacteria and fungi. Both are vital members of the rhizosphere but differ considerably in their life history and dispersal potentiality. Edaphic factors drove the parallel variations of bacterial and fungal communities. Although bacterial and fungal communities exhibited similar biogeographic patterns, the assembly of fungi was more driven by dispersal limitation than selection compared with bacteria. This finding supported the ‘size-dispersal’ hypothesis. pH and total nitrogen respectively mediated the relative importance of deterministic and stochastic processes in shaping bacterial and fungal communities. In addition, fungal communities exhibited potentially broader environmental thresholds and more modular co-occurrence patterns than bacteria (bacteria: 0.67; fungi: 0.78). These results emphasized the importance of dispersal limitation in structuring rhizosphere microbiota and shaping community features of ecologically distinct microorganisms. This study provides insights into the improved prediction and management of the key functions of rhizosphere microbiota.

Abstract Background Magnolia officinalis Rehd. et Wils, commonly called Houpo, has been used for thousands of years in China as a traditional herbal medicine. The primary processing of Houpo requires sweating treatment, which is a special drying process and is considered to be an essential embodiment of high quality and genuine medicinal materials. The sweating of Houpo leads to peculiar changes in the microbial community structure and the content of main active substances (magnolol, honokiol, syringin and magnoflorine). Variation in the microbial community was considered the cause of the change in content of active substances of Houpo, although the microbial taxa responsible for the improvement of content remain unidentified. Methods In this study, we used MiSeq high-throughput sequencing methods for partial bacterial 16S rRNA and 18S rRNA gene sequences to compare the bacterial and fungal community structures at different timepoints in the process of sweating. The content of the main active substances (magnolol, honokiol, syringin and magnoflorine) were determined by high-performance liquid chromatography analysis to evaluate the effects of sweating. UPLC-Q-Extractive Orbitrap mass spectrometry (UPLC-QE Orbitrap MS) was used to detection of differential metabolites of unsweated Houpo before and after co-culture with core bacterial solutions. Results In this study, the total contents of magnolol (MG) and honokiol (HK) were significantly increased at 4 dp (dp for day PM sample), up to 3.75%, and the contents of syringin (SG) and magnoflorine (MF) were as high as 0.12% and 0.06%, respectively. Bacterial abundance and diversity were higher in the early stage (0 day–2 da; da for day AM sample) than in the later stage (4–5 dp), while fungal abundance was more obvious in the later stage than in the early stage. Positive correlation coefficients revealed that the relative abundance of Enterobacter (P

Abstract Background Panax ginseng is widely used as functional food and traditional Chinese medicine. To satisfy the market supply and medication safety, biofertilizers are used as agents to stimulate the growth and production of P. ginseng. Methods In this study, we used high-throughput sequencing and quantitative polymerase chain reaction to analyze microbial community in soils treated with biofertilizers during the development stages of P. ginseng. Ginsenoside content was detected using high-performance liquid chromatography analysis to evaluate the effects of biofertilizer application. Results In this study, the incidence rate of P. ginseng root rot significantly declined by 40.3–47.3% after application of disease-biocontrol biofertilizers. Bacterial diversity showed increasing trends in soils treated with 3.0–4.5 ml kg−1 of disease-biocontrol biofertilizers compared with those in untreated soils. Principal coordinate analysis ordination revealed that bacterial communities were changed by biofertilizers depending on their application concentration. Relative abundance of potentially beneficial bacterial agents, such as Bacillus, Burkholderia, Rhizobium, Streptomyces, and Mycobacterium, significantly increased compared with that in control. Fusarium of low abundance observed in soils treated with biofertilizers compared with that in untreated soils. P. ginseng yield was enhanced by 17.0–19.1%, and ginsenoside (Rg1 and Rb1) contents were improved after biofertilizer application. Conclusions Our results reveal that biofertilizers reduced the incidence rate of root rot, increased bacterial diversity, promoted the relative abundance of potentially beneficial bacterial taxa, decreased the abundance of potentially harmful bacterial agents, and then enhanced the yield and quality of P. ginseng.

In this work, nickel metal supported on different supports (SiO2, Al2O3, ZSM-5) were prepared by spraying nickel nitrate on the supports and calcined at 873 K. Then, they were characterized by XRD, XRF, N2 adsorption–desorption, NH3-TPD, MCH-TPD, H2-TPR, and pyridine-FTIR, and tested as catalysts for the dehydrogenation aromatization and isomerization of methylcyclohexane (MCH) under the conditions of S-Zorb catalytic adsorption desulfurization (T = 673 K, P = 1.5 MPa, WHSV = 5 h−1). The H2-TPR results showed that the interaction of NiO with support decreased in the order of NiO/ZSM-5-Fe

Heavy metal contamination in herbal medicines is a global threat to human beings especially at levels above known threshold concentrations. The concentrations of five heavy metals cadmium (Cd), lead (Pb), arsenic (As), mercury (Hg) and copper (Cu) were investigated using Inductively Coupled Plasma Optical Mass Spectrometry (ICP-MS) with 1773 samples around the world. According to Chinese Pharmacopoeia, 30.51% (541) samples were detected with at least one over-limit metal. The over-limit ratio for Pb was 5.75% (102), Cd at 4.96% (88), As at 4.17% (74), Hg at 3.78% (67), and of Cu, 1.75% (31). For exposure assessment, Pb, Cd, As, and Hg have resulted in higher than acceptable risks in 25 kinds of herbs. The maximal Estimated Daily Intake of Pb in seven herbs, of Cd in five, of Hg in four, and As in three exceeded their corresponding Provisional Tolerable Daily Intakes. In total 25 kinds of herbs present an unacceptable risk as assessed with the Hazard Quotient or Hazard Index. Additionally, the carcinogenic risks were all under acceptable limits. Notably, As posed the highest risk in all indicators including Estimated Daily Intake, Hazard Index, and carcinogenic risks. Therefore further study on enrichment effect of different states of As and special attention to monitoring shall be placed on As related contamination.

Abstract Background Bacterial endophytes are widespread inhabitants inside plant tissues that play crucial roles in plant growth and biotransformation. This study aimed to offer information for the exploitation of endophytes by analyzing the bacterial endophytes in different parts of Panax notoginseng. Methods We used high-throughput sequencing methods to analyze the diversity and composition of bacterial endophytes from different parts of P. notoginseng. Results A total of 174,761 classified sequences were obtained from the analysis of 16S ribosomal RNA in different parts of P. notoginseng. Its fibril displayed the highest diversity of bacterial endophytes. Principal coordinate analysis revealed that the compositions of the bacterial endophytes from aboveground parts (flower, leaf, and stem) differed from that of underground parts (root and fibril). The abundances of Conexibacter, Gemmatimonas, Holophaga, Luteolibacter, Methylophilus, Prosthecobacter, and Solirubrobacter were significantly higher in the aboveground parts than in the underground parts, whereas the abundances of Bradyrhizobium, Novosphingobium, Phenylobacterium, Sphingobium, and Steroidobacter were markedly lower in the aboveground parts. Conclusions Our results elucidated the comprehensive diversity and composition profiles of bacterial endophytes in different parts of 3-year-old P. notoginseng. Our data offered pivotal information to clarify the role of endophytes in the production of P. notoginseng and its important metabolites.

Panax notoginseng is famous for its important therapeutic effects. Saponins are bioactive compounds found in different parts and developmental stages of P. notoginseng plants. Thus, it is urgently to study saponins distribution in different parts and growth ages of P. notoginseng plants. In this study, potential biomarkers were found, and their chemical characteristic differences were revealed through metabolomic analysis. High-performance liquid chromatography data indicated the higher content of saponins (i.e., Rg1, Re, Rd, and Rb1) in the underground parts than that in the aerial parts. 20(S)-Protopanaxadiol saponins were mainly distributed in the aerial parts. Additionally, the total saponin content in the 3-year-old P. notoginseng plant (188.0 mg/g) was 1.4-fold higher than that in 2-year-old plant (130.5 mg/g). The transcriptomic analysis indicated the tissue-specific transcription expression of genes, namely, PnFPS, PnSS, PnSE1, PnSE2, and PnDS, which encoded critical synthases in saponin biosyntheses. These genes showed similar expression patterns among the parts of P. notoginseng plants. The expression levels of these genes in the flowers and leaves were 5.2fold higher than that in the roots and fibrils. These results suggested that saponins might be actively synthesized in the aerial parts and transformed to the underground parts. This study provides insights into the chemical and genetic characteristics of P. notoginseng to facilitate the synthesis of its secondary metabolites and a scientific basis for appropriate collection and rational use of this plant. KEY WORDS: Panax notoginseng, Saponin, Growth years, Metabolomic analyses, Gene expression

The cultivation of Panax plants is hindered by replanting problems, which may be caused by plant-driven changes in the soil microbial community. Inoculation with microbial antagonists may efficiently alleviate replanting issues. Through high-throughput sequencing, this study revealed that bacterial diversity decreased, whereas fungal diversity increased, in the rhizosphere soils of adult ginseng plants at the root growth stage under different ages. Few microbial community, such as Luteolibacter, Cytophagaceae, Luteibacter, Sphingomonas, Sphingomonadaceae, and Zygomycota, were observed; the relative abundance of microorganisms, namely, Brevundimonas, Enterobacteriaceae, Pandoraea, Cantharellales, Dendryphion, Fusarium, and Chytridiomycota, increased in the soils of adult ginseng plants compared with those in the soils of 2-year-old seedlings. Bacillus subtilis 50-1, a microbial antagonist against the pathogenic Fusarium oxysporum, was isolated through a dual culture technique. These bacteria acted with a biocontrol efficacy of 67.8%. The ginseng death rate and Fusarium abundance decreased by 63.3% and 46.1%, respectively, after inoculation with B. subtilis 50-1. Data revealed that microecological degradation could result from ginseng-driven changes in rhizospheric microbial communities; these changes are associated with the different ages and developmental stages of ginseng plants. Biocontrol using microbial antagonists alleviated the replanting problem. KEY WORDS: Panax ginseng, Microbial communities, Replanting problem, High-throughput sequencing, Different ages, Bioremediation

Hydroxypyridinones (HOPOs) have been used in the chelation therapy of iron and actinide metals. Their application in metal-based radiopharmaceuticals has also been increasing in recent years. This review article focuses on how multidentate HOPOs can be used in targeted radiometal-based diagnostic and therapeutic radiopharmaceuticals. The general structure of radiometal-based targeted radiopharmaceuticals, a brief description of siderophores, the basic structure and properties of bidentate HOPO, some representative HOPO multidentate chelating agents, radiopharmaceuticals based on HOPO multidentate bifunctional chelators for gallium-68, thorium-227 and zirconium-89, as well as the future prospects of HOPO multidentate bifunctional chelators in other metal-based radiopharmaceuticals are described and discussed in turn. The HOPO metal-based radiopharmaceuticals that have shown good prospects in clinical and preclinical studies are gallium-68, thorium-227 and zirconium-89 radiopharmaceuticals. We expect HOPO multidentate bifunctional chelators to be a very promising platform for building novel targeted radiometal-based diagnostic and therapeutic radiopharmaceuticals.

Abstract Background American ginseng (Panax quinquefolius L.) is renowned worldwide for its eutherapeutic effects. The replantation of American ginseng usually fails due to problems associated with continuous cropping. An imbalance in the microbial community is thought to be responsible for this, but the overall changes in microbial communities under a continuous cropping system are unclear. Methods This study used quantitative polymerase chain reaction combined with high-throughput sequencing methods to confirm changes in a microbial community under continuous cropping of American ginseng. Results Copy numbers of bacteria and fungi significantly declined by 47.7 and 45.5%, respectively, upon American ginseng cropping over 3 years. A total of 66,391 classified sequences were obtained from high-throughput sequencing analyses of 16S and 18S rRNA in six soil samples. A decline in bacterial diversity and an increase in fungal diversity were observed in the continuous cropping soils of American ginseng compared to those of traditional crops. Compared with soils used for traditional crops, the relative abundance of bacterial and fungal groups changed in soils subjected to continuous cropping with American ginseng. Conclusions Our results revealed that the diversity and composition of soil bacterial and fungal communities changed in the continuous cropping of American ginseng compared to those of traditional crops. Those data provided comprehensive insight into microbial communities at the agro-ecosystem scale and contributed to the understanding of micro-ecological environments in the rhizosphere of medicinal plants.

Panax notoginseng is famous for its important therapeutic effects and commonly used worldwide. The active ingredients saponins have distinct contents in different tissues of P. notoginseng, and they may be related to the expression of key genes in the synthesis pathway. In our study, high-performance liquid chromatography results indicated that the contents of protopanaxadiol-(Rb1, Rc, Rb2, and Rd) and protopanaxatriol-type (R1, Rg1, and Re) saponins in below ground tissues were higher than those in above ground tissues. Clustering dendrogram and PCA analysis suggested that the below and above ground tissues were clustered into two separate groups. A total of 482 and 882 unigenes were shared in the below and above ground tissues, respectively. A total of 75 distinct expressions of CYPs transcripts (RPKM ≥ 10) were detected. Of these transcripts, 38 and 37 were highly expressed in the below ground and above ground tissues, respectively. RT-qPCR analysis showed that CYP716A47 gene was abundantly expressed in the above ground tissues, especially in the flower, whose expression was 31.5-fold higher than that in the root. CYP716A53v2 gene was predominantly expressed in the below ground tissues, especially in the rhizome, whose expression was 20.1-fold higher than that in the flower. Pearson’s analysis revealed that the CYP716A47 expression was significantly correlated with the contents of ginsenoside Rc and Rb2. The CYP716A53v2 expression was associated with the saponin contents of protopanaxadiol-type (Rb1 and Rd) and protopanaxatriol-type (R1, Rg1, and Re). Results indicated that the expression patterns of CYP716A47 and CYP716A53v2 were correlated with the distribution of protopanaxadiol-type and protopanaxatriol-type saponins in P. notoginseng. This study identified the pivotal genes regulating saponin distribution and provided valuable information for further research on the mechanisms of saponin synthesis, transportation, and accumulation.