Mo-Doped Na4Fe3(PO4)2P2O7/C Composites for High-Rate and Long-Life Sodium-Ion Batteries.

Na4Fe3(PO4)2P2O7/C (NFPP) is a promising cathode material for sodium-ion batteries, but its electrochemical performance is heavily impeded by its low electronic conductivity. To address this, pure-phase Mo6+-doped Na4Fe3-xMox(PO4)2P2O7/C (Mox-NFPP, x = 0, 0.05, 0.10, 0.15) with the Pn21a space group is successfully synthesized through spray drying and annealing methods. Density functional theory (DFT) calculations reveal that Mo6+ doping facilitates the transition of electrons from the valence to the conduction band, thus enhancing the intrinsic electron conductivity of Mox-NFPP. With an optimal Mo6+ doping level of x = 0.10, Mo0.10-NFPP exhibits lower charge transfer resistance, higher sodium-ion diffusion coefficients, and superior rate performance. As a result, the Mo0.10-NFPP cathode offers an initial discharge capacity of up to 123.9 mAh g-1 at 0.1 C, nearly reaching its theoretical capacity. Even at a high rate of 10 C, it delivers a high discharge capacity of 86.09 mAh g-1, maintaining 96.18% of its capacity after 500 cycles. This research presents a new and straightforward strategy to enhance the electrochemical performance of NFPP cathode materials for sodium-ion batteries.

The relationship between prognosis and temporal muscle thickness in 102 patients with glioblastoma.

Temporal muscle thickness measured on 3D MRI has recently been linked to prognosis in glioblastoma patients and may serve as an independent prognostic indicator. This single-center study looked at temporal muscle thickness and prognosis in patients with primary glioblastoma. Overall survival was the major study outcome. For a retrospective analysis from 2010 to 2020, clinical data from 102 patients with glioblastoma at the Department of Oncology Radiotherapy of the First Affiliated Hospital of Dalian Medical University were gathered. Fifty-five cases from 2016 to 2020 contained glioblastoma molecular typing data, of which 45 were IDH wild-type glioblastomas and were analysed separately. TMT was measured on enhanced T1-weighted magnetic resonance images in patients with newly diagnosed glioblastoma.Overall patient survival (OS) was calculated by the Kaplan-Meier method and survival curves were plotted using the log-rank-sum test to determine differences between groups, and multifactorial analyses were performed using a Cox proportional-risk model.The median TMT for 102 patients was 6.775 mm (range: 4.95-10.45 mm). Patients were grouped according to median TMT, and the median overall survival (23.0 months) was significantly longer in the TMT > median group than in the TMT median group (P 0.001; Log-rank test). Analysing 45 patients with IDH wild type alone, the median overall survival (12 months) of patients in the TMT > median group was significantly longer than that of patients in the TMT ≤ median group (8 months) (P < 0.001; Log-rank test).TMT can serve as an independent prognostic factor for glioblastoma.

Deciphering the key factors affecting pesticide residue risk in vegetable ecosystem.

Soil contamination, particularly from pesticide residues, presents a significant challenge to the sustainable development of agricultural ecosystems. Identifying the key factors influencing soil pesticide residue risk and implementing effective measures to mitigate their risks at the source are essential. Here, we collected soil samples and conducted a comprehensive survey among local farmers in the Three Gorges Reserve Area, a major agricultural production region in Southwest China. Subsequently, employing a dual analytical approach combining structural equation modeling (SEM) and random forest modeling (RFM), we examined the effects of various factors on pesticide residue accumulation in vegetable ecosystems. Our SEM analysis revealed that soil characteristics (path coefficient 0.85) and cultivation factor (path coefficient 0.84) had the most significant effect on pesticide residue risk, while the farmer factors indirectly influenced pesticide residues by impacting both cultivation factors and soil characteristics. Further exploration using RFM identified the three most influential factors contributing to pesticide residue risk as cation exchange capacity (CEC) (account for 18.84%), cultivation area (account for 14.12%), and clay content (account for 13.01%). Based on these findings, we carried out experimental trials utilizing Integrated Pest Management (IPM) technology, resulting in a significant reduction in soil pesticide residues and notable improvements in crop yields. Therefore, it is recommended that governmental efforts should prioritize enhanced training for vegetable farmers, promotion of eco-friendly plant protection methods, and regulation of agricultural environments to ensure sustainable development.

Pipersarmenoids, new amide alkaloids from Piper sarmentosum.

A chemical investigation of the aerial parts of Piper sarmentosum resulted in the isolation and identification of 14 amide alkaloids, including three new amide alkaloids, pipersarmenoids A - C (1-3), three new natural amide alkaloids, pipersarmenoids D - F (4-6), and 8 known analogues, N-p-coumaroyltyramine (7), piperlotine C (8), piperlotine D (9), pellitorine (10), sarmentine (11), aurantiamide acetate (12), 1-cinnamoyl pyrrolidine (13) and sarmentamide B (14). Their structures were determined by spectroscopic analysis including HRESIMS and 1D and 2D NMR. The cytotoxicity, neuroinflammation-inhibiting and acetylcholinesterase (AChE) inhibitory activities of those compounds were tested. Compounds 1, 2 and 12 inhibited NO production induced by LPS in BV2 cells with IC50 values of 9.36, 12.53 and 10.77 µM, respectively. Moreover, 1, 2, 7 and 11 showed moderate inhibitory activity on AChE with IC50 values ranging from 37.56 to 48.84 µM.

Dynamic monitoring soft tissue healing via visualized Gd-crosslinked double network MRI microspheres.

By integrating magnetic resonance-visible components with scaffold materials, hydrogel microspheres (HMs) become visible under magnetic resonance imaging(MRI), allowing for non-invasive, continuous, and dynamic monitoring of the distribution, degradation, and relationship of the HMs with local tissues. However, when these visualization components are physically blended into the HMs, it reduces their relaxation rate and specificity under MRI, weakening the efficacy of real-time dynamic monitoring. To achieve MRI-guided in vivo monitoring of HMs with tissue repair functionality, we utilized airflow control and photo-crosslinking methods to prepare alginate-gelatin-based dual-network hydrogel microspheres (G-AlgMA HMs) using gadolinium ions (Gd (III)), a paramagnetic MRI contrast agent, as the crosslinker. When the network of G-AlgMA HMs degrades, the cleavage of covalent bonds causes the release of Gd (III), continuously altering the arrangement and movement characteristics of surrounding water molecules. This change in local transverse and longitudinal relaxation times results in variations in MRI signal values, thus enabling MRI-guided in vivo monitoring of the HMs. Additionally, in vivo data show that the degradation and release of polypeptide (K2 (SL)6 K2 (KK)) from G-AlgMA HMs promote local vascular regeneration and soft tissue repair. Overall, G-AlgMA HMs enable non-invasive, dynamic in vivo monitoring of biomaterial degradation and tissue regeneration through MRI, which is significant for understanding material degradation mechanisms, evaluating biocompatibility, and optimizing material design.

Correlation between gout and dry eye disease.

BACKGROUND: Gout is a common form of inflammatory arthritis that can cause a number of serious complications. Complications are common in patients with gout and complicate their management and disease outcome. The recent literature has reported that an increasing number of gout patients are presenting with dry eye symptoms. However, until now, the link between gout and dry eye disease has not been clearly defined. (It is unclear whether the two conditions simply co-exist, whether there are common risk factors, or whether dry eye disease is a complication of gout.) METHODS: A thorough literature search was conducted in the PubMed database to summarize the most recent information on the correlation between gout and dry eye disease and to explore the potential relationship between the pathogenesis of the two. (Objective: Therefore, in this paper, we review the recent literature on the correlation between gout and dry eye disease and explore the potential association between the pathogenesis of both.) RESULTS: Studies in the last five years have shown a correlation between gout and dry eye, i.e., gout is associated with an increased risk of dry eye. The NLRP3-IL-1ß signaling pathway may be a potential mechanism for the combination of gout and dry eye disease; factors such as high blood uric acid and xanthine oxidase activation in gout patients may aggravate the development of dry eye disease; reducing the use of visual display terminals; reducing or abstaining from alcohol consumption; and moderate coffee intake may effectively prevent gout and dry eye disease. CONCLUSIONS: It is an undisputed fact that many gout patients present with dry eye manifestations that seriously affect the quality of life of gout patients, and early detection and treatment of dry eye in gout patients are crucial.

Engineering amino acid residues of pentacyclic triterpene synthases for improving the activity.

Pentacyclic triterpenoids exhibit a wide range of biological activities which have wide applications in the food, cosmetics, and pharmaceutical industries. High-performance chassis strains have been developed for the production of various pentacyclic triterpenoids, e.g., lupane-type and oleanane-type triterpenoids. The production of common pentacyclic triterpenes and their derivatives is limited by the poor activity of typical pentacyclic triterpene synthases (PTSs). However, a general strategy applicable to typical PTSs is still lacking. As typical pentacyclic triterpenes are derived from the baccharenyl cation, engineering the non-active-site residues in the MXXXXR motif might be beneficial for the catalytic efficiencies of typical PTSs by the stabilization of the baccharenyl cation. Here, we develop a general strategy for improving the activity of typical PTSs. As a proof of concept, the activity of three PTSs such as lupeol synthase, ß-amyrin synthase, and α-amyrin synthases was significantly increased up to 7.3-fold by site-directed saturation mutagenesis. This strategy could be applied to improve the activity of various typical PTSs. KEY POINTS: • The strategy could be applied to typical PTSs for improving the activity. • The catalytic activity of typical PTSs was significantly increased.

Stability evaluation of candidate reference genes for real-time qPCR normalization in Rhyzopertha dominica (Coleoptera: Bostrycidae).

Rhyzopertha dominica is a serious stored grain insect pest around the world. Real-time quantitative polymerase chain reaction (RT-qPCR) is a widely used experimental method in molecular biology for detecting the expression of target genes. As appropriate reference genes are essential for normalizing gene expression, the selection of suitable reference genes is the basis of RT-qPCR experiments. In this study, the expression profiles of 7 candidate reference genes of rps3, rps6, rps13, actin, gadph, tubulin, and 18S rRNA were analyzed under 4 different experimental conditions. The expression stability of candidate genes was evaluated using the ΔCt, GeNorm, BestKeeper, NormFinder, and RefFinder methods. The results revealed that different reference genes were suitable for various experiments. Specifically, rps3 and rps6 were appropriate for the developmental stages and all samples: 18S rRNA and rps13 for temperature-related experiments, actin and rps6 for sex-related experiments, and rps6 and gadph for starvation stress. Our results lay essential groundwork for the normalization of RT-qPCR analyses and contribute to genomic and gene functional research of R. dominica.

Hydrogen Ion Capturing Hydrogel Microspheres for Reversing Inflammaging.

Inflammaging is deeply involved in aging-related diseases and can be destructive during aging. The maintenance of pH balance in the extracellular microenvironment can alleviate inflammaging and repair aging-related tissue damage. In this study, the hydrogen ion capturing hydrogel microsphere (GMNP) composed of mineralized transforming growth factor-ß (TGF-ß) and catalase (CAT) nanoparticles is developed via biomimetic mineralization and microfluidic technology for blocking the NLRP3 cascade axis in inflammaging. This GMNP can neutralize the acidic microenvironment by capturing excess hydrogen ions through the calcium carbonate mineralization layer. Then, the subsequent release of encapsulated TGF-ß and CAT can eliminate both endogenous and exogenous stimulus of NLRP3, thus suppressing the excessive activation of inflammaging. In vitro, GMNP can suppress the excessive activation of the TXNIP/NLRP3/IL-1ß cascade axis and enhance extracellular matrix (ECM) synthesis in nucleus pulposus cells. In vivo, GMNP becomes a sustainable and stable niche with microspheres as the core to inhibit inflammaging and promote the regeneration of degenerated intervertebral discs. Therefore, this hydrogen ion-capturing hydrogel microsphere effectively reverses inflammaging by interfering with the excessive activation of NLRP3 in the degenerated tissues.

Reversible data hiding in encrypted images with multi-prediction and adaptive huffman encoding.

With the rapid development of multimedia technology and the massive accumulation of user data, a huge amount of data is rapidly generated and shared over the network, while the problems of inappropriate data access and abuse persist. Reversible data hiding in encrypted images (RDHEI) is a privacy-preserving method that embeds protected data in an encrypted domain and accurately extracts the embedded data without affecting the original content. However, the amount of embedded data has been one of the major limitations in the performance and application of RDHEI. Currently, the main approaches to improve the capacity of RDHEI are either to increase the overall capacity or to reduce the length of the auxiliary information. In this paper, we propose a novel RDHEI scheme based on multi-prediction and adaptive Huffman encoding. To increase the overall capacity, we propose a multi-prediction, called MED+GAP predictor, to generate the label map data of non-reference pixels prior to image encryption. Then, an adaptive Huffman coding is designed to compress the generated labels in order to reduce the embedding length of the auxiliary information used for the extraction and recovery. Experiments show that the proposed method with MED+GAP predictor and adaptive Huffman coding improves 0.052 bpp, 0.023 bpp, and 0.047 bpp on average over the other state-of-the-art methods on the BOSSBase, BOWS-2, and UCID datasets, respectively, while maintaining security and reversibility.

Loneliness is related to smaller gray matter volumes in ACC and right VLPFC in people with major depression: a UK biobank study.

The anterior cingulate cortex (ACC) and right ventrolateral prefrontal cortex (VLPFC) are thought to have important roles in loneliness (feeling of social isolation/exclusion) experience or regulation and in the pathophysiology of their disturbance in major depressive disorder (MDD). However, the structural abnormalities of these regions and the correlates with loneliness in MDD across the healthy population have not fully been clarified. The study analyzed the link between loneliness and gray matter volumes (GMVs) in the ACC and right VLPFC among 1,005 patients with MDD and 7,247 healthy controls (HCs) using UK Biobank data. Significant reductions in GMV in the right VLPFC were found in MDD males compared to HCs. MDD males also showed a higher association between loneliness and reduced GMVs in the right VLPFC and bilateral ACC than HCs. No such associations were found in MDD females. The findings suggest that loneliness may influence brain structures crucial for emotion experience and regulation, particularly in middle-older aged men with MDD. This highlights the potential adverse effects of loneliness on brain structure in MDD and suggests that social engagement could have a positive impact.

Comparing the Effect of Mechanical Loading on Deep and Superficial Cartilage Using Quantitative UTE MRI.

BACKGROUND: The biomechanical properties of deep and superficial cartilage may be different, yet in vivo MRI validation is required. PURPOSE: To compare the effect of mechanical loading on deep and superficial cartilage in young healthy adults using ultrashort echo time (UTE)-T2* mapping. STUDY TYPE: Prospective, intervention. SUBJECTS: Thirty-one healthy adults (54.8% females, median age = 23 years). FIELD STRENGTH/SEQUENCE: 3-T, PD-FS, and UTE sequences with four echo times (TEs = 0.1, 0.5, 2.8, and 4.0 msec; 0.6 mm isotropic spatial resolution) of the left knee, acquired before and after loading exercise. ASSESSMENT: Quantitative UTE-T2* maps of the entire knee were generated using UTE images of four TEs. In deep and superficial cartilage of patella, medial and lateral femur, medial and lateral tibia cartilage (PC, MFC, LFC, MTC, and LTC), which were segmented manually, cartilage thickness and T2* values before and after loading were measured, extracted, taken averages of, and compared. Scan-rescan repeatability was evaluated. Body weight and body mass index (BMI) data were collected. Physical activity levels were evaluated using International Physical Activity Questionnaire. STATISTICAL TESTS: Paired sample t-tests, paired Wilcoxon Mann-Whitney tests, Pearson and Spearman correlation analyses, Kruskal-Wallis tests with post-hoc Bonferroni correction. A P-value <0.05 was considered statistically significant. RESULTS: The scan-rescan repeatability was good (RMSA-CV < 10%). After exercise, deep cartilage exhibited no significant differences in cartilage thickness (PPC = 0.576, PMTC = 0.991, PMFC = 0.899, PLTC = 0.861, PLFC = 0.290) and T2* values (PPC = 0.914, PMTC = 0.780, PMFC = 0.754, PLTC = 0.327, PLFC = 0.811), which both significantly decreased in superficial PC, MFC, LFC, and MTC. The T2* values of superficial MTC and deep MFC were moderately correlated with higher body weight (ρ = 0.431) and lower BMI (ρ = -0.499), respectively. DATA CONCLUSION: Deep and superficial cartilage may respond differently to mechanical loading as assessed by UTE-T2*. EVIDENCE LEVEL: 2 TECHNICAL EFFICACY: Stage 3.

Succession of endophytic fungi and rhizosphere soil fungi and their correlation with secondary metabolites in Fagopyrum dibotrys.

Golden buckwheat (Fagopyrum dibotrys, also known as F. acutatum) is a traditional edible herbal medicinal plant with a large number of secondary metabolites and is considered to be a source of therapeutic compounds. Different ecological environments have a significant impact on their compound content and medicinal effects. However, little is known about the interactions between soil physicochemical properties, the rhizosphere, endophytic fungal communities, and secondary metabolites in F. dibotrys. In this study, the rhizosphere soil and endophytic fungal communities of F. dibotrys in five different ecological regions in China were identified based on high-throughput sequencing methods. The correlations between soil physicochemical properties, active components (total saponins, total flavonoids, proanthocyanidin, and epicatechin), and endophytic and rhizosphere soil fungi of F. dibotrys were analyzed. The results showed that soil pH, soil N, OM, and P were significantly correlated with the active components of F. dibotrys. Among them, epicatechin, proanthocyanidin, and total saponins were significantly positively correlated with soil pH, while proanthocyanidin content was significantly positively correlated with STN, SAN, and OM in soil, and total flavone content was significantly positively correlated with P in soil. In soil microbes, Mortierella, Trechispora, Exophiala, Ascomycota_unclassified, Auricularia, Plectosphaerella, Mycena, Fungi_unclassified, Agaricomycetes_unclassified, Coprinellus, and Pseudaleuria were significantly related to key secondary metabolites of F. dibotrys. Diaporthe and Meripilaceae_unclassified were significantly related to key secondary metabolites in the rhizome. This study presents a new opportunity to deeply understand soil-plant-fungal symbioses and secondary metabolites in F. dibotrys, as well as provides a scientific basis for using biological fertilization strategies to improve the quality of F. dibotrys.

The Biological Roles of Puccinia striiformis f. sp. tritici Effectors during Infection of Wheat.

Puccinia striiformis f. sp. tritici (Pst) is the causative agent of wheat stripe rust, which can lead to a significant loss in annual wheat yields. Therefore, there is an urgent need for a deeper comprehension of the basic mechanisms underlying Pst infection. Effectors are known as the agents that plant pathogens deliver into host tissues to promote infection, typically by interfering with plant physiology and biochemistry. Insights into effector activity can significantly aid the development of future strategies to generate disease-resistant crops. However, the functional analysis of Pst effectors is still in its infancy, which hinders our understanding of the molecular mechanisms of the interaction between Pst and wheat. In this review, we summarize the potential roles of validated and proposed Pst effectors during wheat infection, including proteinaceous effectors, non-coding RNAs (sRNA effectors), and secondary metabolites (SMs effectors). Further, we suggest specific countermeasures against Pst pathogenesis and future research directions, which may promote our understanding of Pst effector functions during wheat immunity attempts.

A Novel Cuticular Protein-like Cpr21L Is Essential for Nymph Survival and Male Fecundity in the Brown Planthopper.

Cuticular proteins (CPs) are a large family and perform a variety of functions. However, the physiological roles of cuticle protein 21-like (Cpr21L) in the brown planthopper (Nilaparvata lugens, BPH), one of the most destructive insect pests of rice, are largely unclear. In this study, Cpr21L was revealed to be expressed in both BPH nymphs and adults, and the mRNA expression level was much higher in male adults than female adults. Spatially, the expression of Cpr21L in the testis was higher than in the ovary. The RNA interference (RNAi) of Cpr21L seriously decreased nymph survival, and no individual survived 8 days post-dsCpr21L injection. The RNAi of Cpr21L in adults also decreased the fertility of males, especially in the dsCpr21L♂ × dsGFP♀ group. The average number of eggs laid by one female in this group significantly decreased by 50.1%, and the eggs' hatchability decreased from 76.5% to 23.8% compared with the control (dsGFP♂ × dsGFP♀). Furthermore, observations under a stereomicroscope showed that the RNAi of Cpr21L severely impaired the development of the testes. Therefore, Cpr21L is essential for the nymphal survival and male fecundity of BPH, thus providing a possible target for pest control.

Metal-organic frameworks doped with metal ions for efficient sterilization: Enhanced photocatalytic activity and photothermal effect.

Photocatalytic and photothermal disinfection is a promising strategy for addressing the challenges of environmental microbial contamination. In this work, we choose a metal-organic framework (MOF), ZIF-8, as an inexpensive and ideal model for metal ion doping, and manipulate the band structure, thermal vibration in molecules, charge distribution, and robustness of the metal-ligand coordination bond of the metal-ion-doped ZIFs for their use in photo-disinfection. The effects of their absorption edge, rate of the photo-induced temperature rise, transient photocurrent response, photo-generated reactive oxygen species (ROS) type, and crystal stability on the photo-disinfection performance are systematically studied by varying the metal ion type (Co2+, Ni2+, or Cu2+) and doping concentration (1-100%). The results show that the efficiency of light harvesting and photogenerated carrier separation is facilitated in all doped ZIFs. The photothermal conversion gradually improves with the increasing concentration of doped Co2+/Cu2+. Remarkably, the photo-generated ROS type changes from the original singlet oxygen (1O2) to multiple ROS (1O2 and •O2-) due to the introduction of Co(II) sites. Consequently, compared with pristine ZIF-8 and other doped ZIFs, Co2+-doped ZIF-8 with a 5% doping concentration shows the highest sterilization efficiency (6.6 log10 CFU mL-1) against Escherichia coli (E. coli) under simulated sunlight within one hour by virtue of the enhanced photothermal effect and the generation of multiple ROS. This work provides insights into the application of metal-ion-doped MOF photocatalysts to the disinfection of environments with pathogenic microorganisms.

In-situelectrochemical fabrication of Ag@AgCl NW-PET film with superior photocatalytic bactericidal activity.

Constructing a composite photocatalyst with efficient charge-transfer pathways is contribute to improving charge separation, which has attracted wide attention owing to its availability in photocatalysis applications. In this work, three-dimensional (3D) silver@silver chloride (Ag@AgCl) network structures are fabricated for photocatalytic inactivation ofEscherichia coli(E. coli) by thein situelectrochemical introducing AgCl shell on the surface of Ag nanowire (NW) networks that are coated on a polyethylene terephthalate (PET) substrate. The obtained Ag@AgCl NW-PET films exhibit good photocatalytic bactericidal activity againstE. coliunder simulated Sunlight irradiation, mainly due to their efficient charge-transport channel constructed by the Ag NWs network. It is worth noting that the content of converted AgCl shell is positively correlated with their photocatalytic bactericidal efficiency. The experimental results also demonstrate that the synergistic contribution of Ag+sustained release, rough surfaces and energy band structure optimization in photocatalytic sterilization. Besides, the prepared Ag@AgCl NW-PET film can be recycled, and the photocatalytic sterilization efficiency can still keep above 99% after three cycles. This work might provide new and more diverse opportunities for the development of excellent charge-transport, recyclable photocatalysts for photocatalytic sterilization.

The complete chloroplast genome of the medical plant Huperzia crispata from the Huperziaceae family: structure, comparative analysis, and phylogenetic relationships.

BACKGROUND: Huperzia crispata, belonging to the Huperziaceae family, is one of the most essential resources of huperzine A for candidate drugs to treat Alzheimer's diseases. However, there is very limited information about H. crispat, and its taxonomic status and interspecific relationships between Huperzia species are still unclear. To investigate the taxonomic classification of Huperzia species and identify species discrimination markers, the complete chloroplast (cp) genome of H. crispata was sequenced and characterized for the first time. METHODS AND RESULTS: Total genomic DNA was isolated and sequenced using the next-generation Illumina NovaSeq 6000 platform. The data were filtered, assembled and annotated by a series software and web service. The results were as follows: the cp genome of H. crispata was 154,320 bp long with a large single-copy (LSC) region of 104,023 bp, a small single-copy (SSC) region of 19,671 bp, and a pair of inverted repeat (IRa and IRb) regions of 15,313 bp. A total of 131 genes, including 87 protein-coding genes, 36 transfer RNA genes (tRNAs), and eight ribosome RNA genes (rRNAs), were annotated in the cp genome. The contraction and expansion of the inverted repeat (IR) regions were relatively conserved in the Huperzia genus. Codon usage bias analysis showed that the encoding rate at the 3-end of codon A/T (74.34%) was significantly higher than that of C/G (25.66%). A total of 8 hotspot loci with high Pi values (> 0.06) were identified in the four Huperzia species based on nucleic acid diversity analysis. Ka/Ks selective pressure analysis demonstrated that the cemA gene is the most common gene undergoing positive selection among Huperzia. In addition, a total of 261 simple sequence repeats and 179 interspersed repeats were identified in the cp genome. Phylogenetic tree analysis based on the complete protein sequences of 23 related species of H. crispata indicated that H. serrata f. longipetiolata is a sister of H. crispata, suggesting that H. serrata f. longipetiolata and H. crispata are more closely related than H. serrata and H. lucidula. CONCLUSIONS: The results strongly supported that H. crispata was more closely related to H. serrata f. longipetiolata than to H. serrata and H. lucidula within the Huperzia genus. The outcome provided important information for the phylogenetic analysis of the subsequent specific molecular species identification in Huperzia. The present results will provide valuable information for further research into the classification, phylogeny and species identification of Huperzia plants.

Engineering Critical Amino Acid Residues of Lanosterol Synthase to Improve the Production of Triterpenoids in Saccharomyces cerevisiae.

Triterpenoids are a subgroup of terpenoids and have wide applications in the food, cosmetics, and pharmaceutical industries. The heterologous production of various triterpenoids in Saccharomyces cerevisiae, as well as other microbes, has been successfully implemented as these production hosts not only produce the precursor of triterpenoids 2,3-oxidosqualene by the mevalonate pathway but also allow simple expression of plant membrane-anchored enzymes. Nevertheless, 2,3-oxidosqualene is natively converted to lanosterol catalyzed by the endogenous lanosterol synthase (Erg7p), causing low production of recombinant triterpenoids. While simple deletion of ERG7 was not effective, in this study, the critical amino acid residues of Erg7p were engineered to lower this critical enzyme activity. The engineered S. cerevisiae indeed accumulated 2,3-oxidosqualene up to 180 mg/L. Engineering triterpenoid synthesis into the ERG7-modified strain resulted in 7.3- and 3-fold increases in the titers of dammarane-type and lupane-type triterpenoids, respectively. This study presents an efficient inducer-free strategy for lowering Erg7p activity, thereby providing 2,3-oxidosqualene for the enhanced production of various triterpenoids.