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Understanding genome-wide diversity, inbreeding, and the burden of accumulated deleterious mutations in small and isolated populations is essential for predicting and enhancing population persistence and resilience. However, these effects are rarely studied in limestone karst plants. Here, we re-sequenced the nuclear genomes of 62 individuals of the Begonia masoniana complex (B. liuyanii, B. longgangensis, B. masoniana and B. variegata) and investigated genomic divergence and genetic load for these four species. Our analyses revealed four distinct clusters corresponding to each species within the complex. Notably, there was only limited admixture between B. liuyanii and B. longgangensis occurring in overlapping geographic regions. All species experienced historical bottlenecks during the Pleistocene, which were likely caused by glacial climate fluctuations. We detected an asymmetric historical gene flow between group pairs within this timeframe, highlighting a distinctive pattern of interspecific divergence attributable to karst geographic isolation. We found that isolated populations of B. masoniana have limited gene flow, the smallest recent population size, the highest inbreeding coefficients, and the greatest accumulation of recessive deleterious mutations. These findings underscore the urgency to prioritize conservation efforts for these isolated population. This study is among the first to disentangle the genetic differentiation and specific demographic history of karst Begonia plants at the whole-genome level, shedding light on the potential risks associated with the accumulation of deleterious mutations over generations of inbreeding. Moreover, our findings may facilitate conservation planning by providing critical baseline genetic data and a better understanding of the historical events that have shaped current population structure of rare and endangered karst plants.

The widespread dissemination of the COVID-19 pandemic precipitated a transition from traditional, in-person pedagogy to online methods within higher vocational education. The inherent limitations of online learning have become progressively conspicuous. Given its pivotal role amidst the pandemic, numerous institutions have endeavored to integrate technology with traditional classroom pedagogies. Consequently, blended learning has emerged as a focal point within vocational education, garnering notable attention and interest. The question arises: which learning method proves most effective for vocational skills development? This study endeavored to ascertain the comparative effectiveness of various learning methods for vocational skills training. Specifically, employing a quantitative approach, the study conducted a comprehensive questionnaire survey of graduates of higher vocational institutions in China. Participants have undergone training via exclusively physical, thoroughly online, and blended learning formats, respectively. The objective was to evaluate the efficacy of these three methods as perceived by higher vocational graduates. In addition, the study aimed to analyze the personal and pedagogical factors that influence the effectiveness of these learning methods. The collected data underwent analysis utilizing SPSS software. One-way analysis of variance (ANOVA) tests, correlation analyses, and subsequent least significant difference (LSD) post-hoc analyses were employed to investigate the correlations between demographic variables and factors related to teachers and educational resources across the three learning methods. These findings indicate that blended learning is the most efficient approach, with a notable preference among vocational graduates. Physical learning demonstrated superior effectiveness, particularly in functional skills training. In divergence, online learning ranked the lowest among graduates' preference, efficiency, and efficacy in functional skills training. Among the demographic factors examined, gender and duration of graduation exhibited notable variances across the effectiveness of different learning methods. However, there are no significant differences in the area of origin, economic region, type of higher vocational institution, and majors. These insights provide valuable guidance for higher vocational institutions in selecting the most suitable learning methods for enhancing functional skills training.

Abstract Euphorbia, one of the largest genera of flowering plants, is well-known for containing many biofuel crops. Euphorbia tirucalli, an evergreen succulent mainly native to the Africa continent but cultivated worldwide, is a promising petroleum plant with high tolerance to drought and salt stress. However, the exploration of such an important plant resource is severely hampered by the lack of a reference genome. Here, we present the chromosome-level genome assembly of E. tirucalli using PacBio HiFi sequencing and Hi-C technology. Its genome size was approximately 745.62 Mb, with a contig N50 of 74.16 Mb. A total of 743.63 Mb (99.73%) of the assembled sequences were anchored to 10 chromosomes with a complete BUSCO score of 97.80%. Genome annotation revealed 26,304 protein-coding genes, and 76.37% of the genome was identified as repeat elements. The high-quality genome provides valuable genetic resources that would be useful for unraveling the genetic mechanisms of biofuel synthesis and evolutionary adaptation of E. tirucalli.

Objective Knee kinematic asymmetries after anterior cruciate ligament reconstruction (ACLR) are correlated with poor clinical outcomes, such as the progression of knee cartilage degenerations or reinjuries. Fast walking in patients with knee conditions may exacerbate knee kinematic asymmetries, but its impact on ACLR patients is uncertain. The aim of this study is to investigate if fast walking induces more knee kinematic asymmetries in unilateral ACLR patients. Methods This cross‐sectional study enrolled 55 patients with unilateral ACLR from January 2020 to July 2022. There were 48 males and seven females with an average age of 30.6 ± 6.4 years. Knee kinematic data were collected at three walking speeds: self‐selected, fast (150% normal), and slow (50% normal). A 3D knee kinematic analysis system measured the data, and self‐reported outcomes assessed comfort levels during walking. We used SPM1D for two‐way repeated ANOVA and posthoc paired t‐tests to analyze kinematic differences in groups. Results In fast walking, ACLR knees exhibited more transverse kinematic asymmetries than intact knees, including greater external rotation angle (1.8°, 38%–43%; gait cycle [GC], p

Introduction: The Maximum Entropy models (MaxEnt) is commonly employed for early detection of disease transmission, particularly effective in assessing the risk zones and intensity of tick-borne disease transmission based on climatic factors.Methods: Utilizing the diversity data of common ticks in Qinghai, along with surveillance statistics tick-borne pathogens of ticks and epidemiological information, we have charted and predicted the prevalence of tick and tick-borne pathogens across Qinghai province.Results and Discussion: The findings indicate that the pivotal environmental factors influencing the distribution of ticks and tick-borne pathogens include land use and land cover (Lucc), elevation (Elev), annual precipitation (Bio12) and annual mean temperature (Bio1). Notably, for Dermacentor nuttalli, the significant environmental factors accounted for 35.2% for Lucc, 24.7% for Elev and 18.6% for Bio12. In the case of Dermacentor silvaru, the pivotal factors contributed 33.2% for Lucc, 23.7% for Elev and 22.6% for Bio1. For Haemaphysalis qinghaiensis, the key environmental variables were 45% for Elev, 30.9% for Lucc and 18.4% for Bio12. Regarding the pathogens, the environmental factors influencing Borrelia burgdorferi contributed 61.5% for Lucc, 13.3% for Elev and 11.9% for Bio1. For Piroplasmida, the contribution was 62.1% for Lucc, 16.7% for Bio1 and 9.9% for Bio12. Rickettsia was influenced by factors accounting for 34.2% for Lucc, 29.7% for Elev and 17.3% for Bio12, while Anaplasma variables contributed 38.2% for Bio1, 26.6% for Lucc and 18.9% for Bio12. The finding indicated that the three predominantly ticks species (D. nuttalli, D. silvaru, and H. qinghaiensis) and the primary tick-borne pathogens (B. burgdorferi, Piroplasmida, Rickettsia, and Anaplasma) are predominantly concentrated in the source regions of the Yellow River, the Yangtze River, the Lancang River, and the eastern part of the Haixi Autonomous Prefecture. Projected under the ssp245 scenario, there is a notable expansion trend in the risk areas for ticks and tick-borne pathogens These findings are consistent with previous epidemiological studies on major ticks and tick-borne diseases in Qinghai conducted by our laboratory, this suggests the feasibility of using the MaxEnt model to predict the distribution of tick-borne disease transmission and compensates for the paucity of research on the Maxent model in the Qinghai Province.

Background: Ticks are ectoparasites that feed on blood and pose a threat to both the livestock industry and public health due to their ability to transmit pathogens through biting. However, the impact of factors such as bloodmeal and geographic regions on the bacterial microbiota of Haemaphysalis qinghaiensis remains poorly understood. Methods: In this study, we used the v3-v4 region of the 16S rRNA gene to sequence the microbiota of Haemaphysalis qinghaiensis from eight groups (HY_M, YS_M, XH_M, LD_M, BM_M, LD_F_F, LD_F, and BM_F_F) in Qinghai Province. Results: Significant differences in bacterial richness were observed between LD_F_F, BM_F_F, and LD_F (P

Abstract Background Citrus is one of the most valuable fruits worldwide and an economic pillar industry in southern China. Nevertheless, it frequently suffers from undesirable environmental stresses during the growth cycle, which severely restricts the growth, development and yield of citrus. In plants, the growth-regulating factor (GRF) family of transcription factors (TF) is extensively distributed and plays an vital part in plant growth and development, hormone response, as well as stress adaptation. However, the systematic identification and functional analysis of GRF TFs in citrus have not been reported. Results Here, a genome-wide identification of GRF TFs was performed in Citrus sinensis, 9 members of CsGRFs were systematically identified and discovered to be scattered throughout 5 chromosomes. Subsequently, physical and chemical properties, phylogenetic relationships, structural characteristics, gene duplication events, collinearity and cis-elements of promoter were elaborately analyzed. In particular, the expression patterns of the CsGRF genes in response to multiple phytohormone and abiotic stress treatments were investigated. Predicated on this result, CsGRF04, which exhibited the most differential expression pattern under multiple phytohormone and abiotic stress treatments was screened out. Virus-induced gene silencing (VIGS) technology was utilized to obtain gene silenced plants for CsGRF04 successfully. After the three stress treatments of high salinity, low temperature and drought, the CsGRF04-VIGS lines showed significantly reduced resistance to high salinity and low temperature stresses, but extremely increased resistance to drought stress. Conclusions Taken together, our findings systematically analyzed the genomic characterization of GRF family in Citrus sinensis, and excavated a CsGRF04 with potential functions under multiple abiotic stresses. Our study lay a foundation for further study on the function of CsGRFs in abiotic stress and hormone signaling response.

BackgroundLong term hypertension seriously promotes target organ damage in the brain and heart, and has increasingly become serious public health problem worldwide. The anti-hypertensive effects of capsaicin has been reported, however, the role and mechanism of capsaicin within the brain on salt-induced hypertension have yet to be elucidated. This study aimed to verify the hypothesis that capsaicin attenuates salt-induced hypertension via the AMPK/Akt/Nrf2 pathway in hypothalamic paraventricular nucleus (PVN).MethodsDahl salt-sensitive (Dahl S) rats were used as animal model for the present study. Rats were randomly divided into four groups based on their dietary regimen (0.3% normal salt diet and 8% high salt diet) and treatment methods (infusion of vehicle or capsaicin in the PVN). Capsaicin was chronically administered in the PVN throughout the animal experiment phase of the study that lasted 6 weeks.ResultsOur results demonstrated that PVN pretreatment with capsaicin can slow down raise of the blood pressure elevation and heart rate (HR) of Dahl S hypertensive rats given high salt diet. Interestingly, the cardiac hypertrophy was significantly improved. Furthermore, PVN pretreatment with capsaicin induced decrease in the expression of mRNA expression of NADPH oxidase-2 (NOX2), inducible nitric oxide synthase (iNOS), NOX4, p-IKKβ and proinflammatory cytokines and increase in number of positive cell level for Nrf2 and HO-1 in the PVN of Dahl S hypertensive rats. Additionally, the protein expressions of phosphatidylinositol 3-kinase (p-PI3K) and phosphorylated protein kinase-B (p-AKT) were decreased, phosphorylated adenosine monophosphate-activated protein kinase (p-AMPK) were increased after the PVN pretreatment with capsaicin.ConclusionCapsaicin pretreatment attenuates salt-sensitive hypertension by alleviating AMPK/Akt/iNOS pathway in the PVN.

The phyllosphere, particularly the leaf surface of plants, harbors a diverse range of microbiomes that play a vital role in the functioning of terrestrial ecosystems. However, our understanding of microbial successions and their impact on functional genes during plant community development is limited. In this study, considering core and satellite microbial taxa, we characterized the phyllosphere microbiome and functional genes in various microhabitats (i.e., leaf litter, moss and plant leaves) across the succession of a plant community in a low-altitude glacier foreland. Our findings indicate that phyllosphere microbiomes and associated ecosystem stability increase during the succession of the plant community. The abundance of core taxa increased with plant community succession and was primarily governed by deterministic processes. In contrast, satellite taxa abundance decreased during plant community succession and was mainly governed by stochastic processes. The abundance of microbial functional genes (such as C, N, and P hydrolysis and fixation) in plant leaves generally increased during the plant community succession. However, in leaf litter and moss leaves, only a subset of functional genes (e.g., C fixation and degradation, and P mineralization) showed a tendency to increase with plant community succession. Ultimately, the community of both core and satellite taxa collaboratively influenced the characteristics of phyllosphere nutrient-cycling genes, leading to the diverse profiles and fluctuating abundance of various functional genes during plant community succession. These findings offer valuable insights into the phyllosphere microbiome and plant–microbe interactions during plant community development, advancing our understanding of the succession and functional significance of the phyllosphere microbial community.

The widespread use of copper and tetracycline as growth promoters in the breeding industry poses a potential threat to environmental health. Nevertheless, to the best of our knowledge, the potential adverse effects of copper and tetracycline on the gut microbiota remain unknown. Herein, mice were fed different concentrations of copper and/or tetracycline for 6 weeks to simulate real life–like exposure in the breeding industry. Following the exposure, antibiotic resistance genes (ARGs), potential pathogens, and other pathogenic factors were analyzed in mouse feces. The co-exposure of copper with tetracycline significantly increased the abundance of ARGs and enriched more potential pathogens in the gut of the co-treated mice. Copper and/or tetracycline exposure increased the abundance of bacteria carrying either ARGs, metal resistance genes, or virulence factors, contributing to the widespread dissemination of potentially harmful genes posing a severe risk to public health. Our study provides insights into the effects of copper and tetracycline exposure on the gut resistome and potential pathogens, and our findings can help reduce the risks associated with antibiotic resistance under the One Health framework.

Abstract Prunus campanulata is an important flowering cherry germplasm of high ornamental value. Given its early-flowering phenotypes, P. campanulata could be used for molecular breeding of ornamental species and fruit crops belonging to the subgenus Cerasus. Here, we report a chromosome-scale assembly of P. campanulata with a genome size of 282.6 Mb and a contig N50 length of 12.04 Mb. The genome contained 24,861 protein-coding genes, of which 24,749 genes (99.5%) were functionally annotated, and 148.20 Mb (52.4%) of the assembled sequences are repetitive sequences. A combination of genomic and population genomic analyses revealed a number of genes under positive selection or accelerated molecular evolution in P. campanulata. Our study provides a reliable genome resource, and lays a solid foundation for genetic improvement of flowering cherry germplasm.

Abstract Background Leeches are an integral component of aquatic biocenosis and can be found in a wide range of ecosystems such as freshwater, saltwater, flowing, and still-water ecosystems. It especially plays an important role in the freshwater benthic community and is an important part of the food web. In this study, a leech species was found in the mantle cavity of wild freshwater mussels in Zigong City, Sichuan Province, China, and its identity was determined through morphological analysis and molecular biological analysis. Results The leech is Hemiclepsis khankiana, a new species of Hemiclepsis that has been discovered in Russia in recent years. Through morphological analysis, the current survey observed that the morphological characteristics of Hemiclepsis khankiana eyespots were significantly different from the first reported description. The first pair of eyespots on the leech were separated and clear, while it had been reduced to unclear shadows in the previous report. The phylogenetic tree based on the COI gene showed that the COI gene sequence obtained in this study was in the same evolutionary branch as Hemiclepsis khankiana (MN295420, MN295421). Genetically, it was most closely related to Hemiclepsis kasmiana (mean COI p-distance = 3.98%). Conclusions The current study reported on the new distribution range of Hemiclepsis khankiana, which was initially discovered in China. This study indicates that the distribution range of the leech species has expanded, laying a foundation for further studies in China.

Multilayer monolithic zirconia, which incorporates polychromatic layers that mimic natural tooth gradients, offers enhanced aesthetics and functionality while reducing debonding risks and improving fabrication efficiency. However, uncertainties remain regarding the fracture characteristics of multilayer monolithic zirconia crowns under occlusal loading, whether composed of uniform or combined yttria levels. The current study investigated how variations in yttria levels and thicknesses affected the optical properties and fracture loads of multilayer monolithic zirconia. Samples of multilayer monolithic zirconia in the Vita A1 shade were used, while employing 3Y (SZ) and 4Y + 5Y (AZ) yttria levels. The optical properties, including the color difference (ΔEWS) and translucency parameters (TP00), were measured using a digital colorimeter. The fracture loads were analyzed using a universal testing machine, and fractured surfaces were examined under a stereomicroscope. Statistical analyses assessed the impacts of the yttria levels and sample thicknesses on the optical properties. The ΔEWS values of SZ ranged 3.6 to 4.0, while for AZ, ΔEWS at 0.5 mm was 3.9 and TP00 values decreased with an increased thickness, with AZ generally exhibiting greater translucency than SZ. In the fracture load investigations, SZ (>1600 N) generally exceeded AZ (>1260 N), with fracture loads notably increasing with thickness, particularly for premolars (SZ > 3270 N, AZ > 2257 N). SZ predominantly exhibited partial and complete fractures, whereas AZ showed fewer non-fracture categorizations. Complete fractures began with dense, irregular cracks that extended outward to reveal smooth surfaces, while premolars subjected to higher loads exhibited concentric ripple-like structures. Partial fractures revealed radial textures indicative of areas of stress concentration. In summary, higher yttria levels were correlated with increased translucency, while variations in the fracture loads primarily stemmed from differences in the tooth position or thickness. Overall, multilayer monolithic zirconia incorporating combined yttria levels of 4Y + 5Y (AZ) offered high translucency, precise color matching, and substantial fracture resistance, rendering it highly suitable for aesthetic and functional dental applications.

Background: Puerarin is an isoflavone compound isolated from the roots of a leguminous plant, the wild kudzu. Various functional activities of this compound in multiple diseases have been reported. However, the effect and mechanism of puerarin in improving blood pressure remain non-elucidated. Purpose: The current study was designed to assess the preventive effects of puerarin on the onset and progression of hypertension and to verify the hypothesis that puerarin alleviates blood pressure by inhibiting the ROS/TLR4/NLRP3 inflammasome signaling pathway in the hypothalamic paraventricular nucleus (PVN) of salt-induced prehypertensive rats. Methods: Male Dahl salt-sensitive rats were fed low NaCl salt (3% in drinking water) for the control (NS) group or 8% (HS) to induce prehypertension. Each batch was divided into two group and treated by bilateral PVN microinjection with either artificial cerebrospinal fluid or puerarin through a micro-osmotic pump for 6 weeks. The mean arterial pressure (MAP) was recorded, and samples were collected and analyzed. Results: We concluded that puerarin significantly prevented the elevation of blood pressure and effectively alleviated the increase in heart rate caused by high salt. Norepinephrine (NE) in the plasma of salt-induced prehypertensive rats also decreased upon puerarin chronic infusion. Additionally, analysis of the PVN sample revealed that puerarin pretreatment decreased the positive cells and gene level of TLR4 (Toll-like receptor 4), NLRP3, Caspase-1 p10, NOX2, MyD88, NOX4, and proinflammatory cytokines in the PVN. Puerarin pretreatment also decreased NF-κBp65 activity, inhibited oxidative stress, and alleviated inflammatory responses in the PVN. Conclusion: We conclude that puerarin alleviated blood pressure via inhibition of the ROS/TLR4/NLRP3 inflammasome signaling pathway in the PVN, suggesting the therapeutic potential of puerarin in the prevention of hypertension.

The dynamic characterization of guest molecules in the metal–organic frameworks (MOFs) can always provide the insightful and inspiring information to facilitate the synthetic design of MOF materials from the bottom-up design of perspective. Herein, we present a series of atomistic molecular dynamics simulation for investigating the bipyridine dicarboxylate (bpydc) linker rotation effect on guest molecule adsorption with and without considering the transition metal (TM) chelation in MOF-253 materials. The simulated PXRD patterns of the various linker orientations present the challenge of distinguishing these structural varieties by the conventional crystalline spectroscopic measurements. The observed short inter-TM stable structure may subsequently lead to the formation of a binuclear TM catalytic site, and a proposed formic acid generation mechanism from CO2 and H2 is derived based upon the density functional theory calculations for the application of CO2 reduction.

Three new Otacilia species were collected from Jiangxi Provinces, China during a survey of the spider fauna of the region: Otacilia anfu Liu, sp. nov. (♂♀), O. guanshan Liu, sp. nov. (♂♀), and O. mingyueshan Liu, sp. nov. (♂♀). All species are described and illustrated with photographs and SEM micrographs, and their distribution is also mapped.

This paper is concerned with piecewise pseudo almost periodic solutions of a class of interval general BAM neural networks with mixed time-varying delays and impulsive perturbations. By adopting the exponential dichotomy of linear differential equations and the fixed point theory of contraction mapping. The sufficient conditions for the existence of piecewise pseudo almost periodic solutions of the interval general BAM neural networks with mixed time-varying delays and impulsive perturbations are obtained. By adopting differential inequality techniques and mathematical methods of induction, the global exponential stability for the piecewise pseudo almost periodic solutions of the interval general BAM neural networks with mixed time-varying delays and impulsive perturbations is discussed. An example is given to illustrate the effectiveness of the results obtained in the paper.

Background: Magnetoencephalography (MEG) is a non-invasive imaging technique for directly measuring the external magnetic field generated from synchronously activated pyramidal neurons in the brain. The optically pumped magnetometer (OPM) is known for its less expensive, non-cryogenic, movable and user-friendly custom-design provides the potential for a change in functional neuroimaging based on MEG. Methods: An array of OPMs covering the opposite sides of a subject’s head is placed inside a magnetically shielded room (MSR) and responses evoked from the auditory cortices are measured. Results: High signal-to-noise ratio auditory evoked response fields (AEFs) were detected by a wearable OPM-MEG system in a MSR, for which a flexible helmet was specially designed to minimize the sensor-to-head distance, along with a set of bi-planar coils developed for background field and gradient nulling. Neuronal current sources activated in AEF experiments were localized and the auditory cortices showed the highest activities. Performance of the hybrid optically pumped magnetometer-magnetoencephalography/electroencephalography (OPM-MEG/EEG) system was also assessed. Conclusions: The multi-channel OPM-MEG system performs well in a custom built MSR equipped with bi-planar coils and detects human AEFs with a flexible helmet. Moreover, the similarities and differences of auditory evoked potentials (AEPs) and AEFs are discussed, while the operation of OPM-MEG sensors in conjunction with EEG electrodes provides an encouraging combination for the exploration of hybrid OPM-MEG/EEG systems.

Statement of problem: The relation between cement color and abutment substrate material and the corresponding effect on the color accuracy of high-transparency pre-colored zirconia (HT-Zr) remains unclear. Purpose: This in-vitro study aimed to investigate the difference in color accuracy when the HT-Zr is bonded to different materials-based substrates with differently colored resin cement. Materials and methods: Vita A1 shade HT-Zr with 1 mm thickness was used as the testing sample. The samples were first placed on zirconia (ZR), tooth color resin (CR), and metallic (MT) abutment substrates. Subsequently, four differently colored cements (translucent (TR), bleach, opaque, and A2 shade (A2)) were used for bonding HT-Zr onto the substrate, and the non-bonded group was used as the control group (CG). There were 15 groups in total (n = 10 per group). A digital colorimeter was used to obtain Commission Internationale de l’Eclairage (CIELab) color parameters. The translucency parameter (TP00) of the substrate and sample, as well as color difference (ΔE00) and chroma (C) between the different groups were calculated. Additionally, the ΔE00 and TP00 were compared with the moderately unacceptable match of ΔE00 = 3.6. The statistical analysis was conducted using ANOVA and Tukey HSD post-hoc test (α = 0.05). Results: HT-Zr exhibited high translucency (TP00 = 11.02 ± 0.18), and the mean ΔE00 of the testing samples ranged between 2.18 ± 0.20 and 13.14 ± 0.31. The ZR-CG and MT-A2 groups showed the highest and lowest lightness separately. The CR-CG group exhibited the highest C, and the ΔE00 was lower than that of 3.6. The MT-TR group showed the lowest C and the highest ΔE00. The inter-group comparison revealed that the ΔE00 for different cement is mostly lower than the acceptable color match of 1.0; moreover, the ΔE00 for all the substrates, excluding the CG group, is higher than 3.6. Conclusions: The abutment substrate materials and the cement color should be considered with caution when using HT-Zr, with the effect of abutment substrate materials being more apparent in color accuracy. HT-Zr restorations are not recommended for discolored or bleached abutments but only for natural-colored abutments to achieve the optimal color appearance.

Abstract Background Paeonia veitchii Lynch, a well-known herb from the Qinghai-Tibet Plateau south of the Himalayas, can synthesize specific monoterpene glycosides (PMGs) with multiple pharmacological activities, and its rhizome has become an indispensable ingredient in many clinical drugs. However, little is known about the molecular background of P. veitchii, especially the genes involved in the biosynthetic pathway of PMGs. Results A corrective full-length transcriptome with 30,827 unigenes was generated by combining next-generation sequencing (NGS) and single-molecule real-time sequencing (SMRT) of six tissues (leaf, stem, petal, ovary, phloem and xylem). The enzymes terpene synthase (TPS), cytochrome P450 (CYP), UDP-glycosyltransferase (UGT), and BAHD acyltransferase, which participate in the biosynthesis of PMGs, were systematically characterized, and their functions related to PMG biosynthesis were analysed. With further insight into TPSs, CYPs, UGTs and BAHDs involved in PMG biosynthesis, the weighted gene coexpression network analysis (WGCNA) method was used to identify the relationships between these genes and PMGs. Finally, 8 TPSs, 22 CYPs, 7 UGTs, and 2 BAHD genes were obtained, and these putative genes were very likely to be involved in the biosynthesis of PMGs. In addition, the expression patterns of the putative genes and the accumulation of PMGs in tissues suggested that all tissues are capable of biosynthesizing PMGs and that aerial plant parts could also be used to extract PMGs. Conclusion We generated a large-scale transcriptome database across the major tissues in P. veitchii, providing valuable support for further research investigating P. veitchii and understanding the genetic information of plants from the Qinghai-Tibet Plateau. TPSs, CYPs, UGTs and BAHDs further contribute to a better understanding of the biology and complexity of PMGs in P. veitchii. Our study will help reveal the mechanisms underlying the biosynthesis pathway of these specific monoterpene glycosides and aid in the comprehensive utilization of this multifunctional plant.

IntroductionWhole-genome resequencing technology covers almost all nucleotide variations in the genome, which makes it possible to carry out conservation genomics research on endangered species at the whole-genome level.MethodsIn this study, based on the whole-genome resequencing data of 101 critically endangered Artocarpus nanchuanensis individuals, we evaluated the genetic diversity and population structure, inferred the demographic history and genetic load, predicted the potential distributions in the past, present and future, and classified conservation units to propose targeted suggestions for the conservation of this critically endangered species.ResultsWhole-genome resequencing for A. nanchuanensis generated approximately 2 Tb of data. Based on abundant mutation sites (25,312,571 single nucleotide polymorphisms sites), we revealed that the average genetic diversity (nucleotide diversity, π) of different populations of A. nanchuanensis was relatively low compared with other trees that have been studied. And we also revealed that the NHZ and QJT populations harboured unique genetic backgrounds and were significantly separated from the other five populations. In addition, positive genetic selective signals, significantly enriched in biological processes related to terpene synthesis, were identified in the NHZ population. The analysis of demographic history of A. nanchuanensis revealed the existence of three genetic bottleneck events. Moreover, abundant genetic loads (48.56% protein-coding genes) were identified in Artocarpus nanchuanensis, especially in genes related to early development and immune function of plants. The predication analysis of suitable habitat areas indicated that the past suitable habitat areas shifted from the north to the south due to global temperature decline. However, in the future, the actual distribution area of A. nanchuanensis will still maintain high suitability.DiscussionBased on total analyses, we divided the populations of A. nanchuanensis into four conservation units and proposed a number of practical management suggestions for each conservation unit. Overall, our study provides meaningful guidance for the protection of A. nanchuanensis and important insight into conservation genomics research.

Background and purpose: Testosterone is an essential sex hormone in maintaining masculine characteristics, which is prescribed for male hypogonadism as testosterone replacement treatment (TRT). Herein, we investigated long-standing controversies about the association between TRT and major adverse cardiovascular events (MACEs), based on real world adverse event (AE) reports, registered in the Food and Drug Administration Adverse Event Reporting System (FAERS).Methods: Publicly available FAERS data from 1 January 2004 to 31 December 2022 were retrieved from the Food and Drug Administration (FDA) website. The data mining protocol including the reporting odds ratio (ROR) and the Bayesian confidence propagation neural network (BCPNN) was applied to analyze overreporting caused by risk factors and MACEs, including TRT, morbidities, and ages. The ROR and the BCPNN were also applied to investigate the annually developing trend of pharmacovigilance (PV) signals in the real world, retrospectively.Results: A total of 3,057 cases referring to MACEs, with a median age of 57 years old (yo), were identified from 28,921 cases of testosterone users. MACEs related to PV signals have emerged since 2014, including cardiac death, non-fatal myocardial infarction, and non-fatal stroke. Myocardial infarction (MI) (ROR: 9.46; IC025: 3.08), acute myocardial infarction (AMI) (ROR: 16.20; IC025: 3.72), ischemic cardiomyopathy (ROR: 11.63; IC025: 2.20), and cardiomyopathy (ROR: 5.98; IC025: 1.96) were the most significant signals generated, and weaker signals included cardiac failure acute (ROR: 4.01; IC025: 0.71), cardiac arrest (ROR: 1.88; IC025: 0.56), and ventricular fibrillation (VF) (ROR: 2.38; IC025: 0.38). The time-to-onset (TTO) of MACEs was calculated with a median of 246 days for AMI.Conclusion: For myocardial infarction and cardiomyopathy, TRT statistically tended to increase the risk of MACEs, while for cardiac arrhythmia, cardiac failure, and stroke, TRT demonstrated beneficial effects among the population with morbidities, such as testosterone deficiency (TD), diabetes mellitus (DM), and hypertension. MACEs were rare but led to serious outcomes including significant increase in death and disability. Since 2018, and before 2014, reports referring to TRT associated with MACEs were relatively scarce, which indicated that there might be a considerable number of cases that went unrecorded, due to neglection. Health workers and testosterone users might pay more attention to testosterone-induced MACEs.