Deep profiling of potential substrate atlas of porcine epidemic diarrhea virus 3C-like protease.

Coronavirus (CoV) 3C-like protease (3CLpro) is essential for viral replication and is involved in immune escape by proteolyzing host proteins. Deep profiling the 3CLpro substrates in the host proteome extends our understanding of viral pathogenesis and facilitates antiviral drug discovery. Here, 3CLpro from porcine epidemic diarrhea virus (PEDV), an enteropathogenic CoV, was used as a model which to identify the potential 3CLpro cleavage motifs in all porcine proteins. We characterized the selectivity of PEDV 3CLpro at sites P5-P4'. We then compiled the 3CLpro substrate preferences into a position-specific scoring matrix and developed a 3CLpro profiling strategy to delineate the protein substrate landscape of CoV 3CLpro. We identified 1,398 potential targets in the porcine proteome containing at least one putative cleavage site and experimentally validated the reliability of the substrate degradome. The PEDV 3CLpro-targeted pathways are involved in mRNA processing, translation, and key effectors of autophagy and the immune system. We also demonstrated that PEDV 3CLpro suppresses the type 1 interferon (IFN-I) cascade via the proteolysis of multiple signaling adaptors in the retinoic acid-inducible gene I (RIG-I) signaling pathway. Our composite method is reproducible and accurate, with an unprecedented depth of coverage for substrate motifs. The 3CLpro substrate degradome establishes a comprehensive substrate atlas that will accelerate the investigation of CoV pathogenicity and the development of anti-CoV drugs.IMPORTANCECoronaviruses (CoVs) are major pathogens that infect humans and animals. The 3C-like protease (3CLpro) encoded by CoV not only cleaves the CoV polyproteins but also degrades host proteins and is considered an attractive target for the development of anti-CoV drugs. However, the comprehensive characterization of an atlas of CoV 3CLpro substrates is a long-standing challenge. Using porcine epidemic diarrhea virus (PEDV) 3CLpro as a model, we developed a method that accurately predicts the substrates of 3CLpro and comprehensively maps the substrate degradome of PEDV 3CLpro. Interestingly, we found that 3CLpro may simultaneously degrade multiple molecules responsible for a specific function. For instance, it cleaves at least four adaptors in the RIG-I signaling pathway to suppress type 1 interferon production. These findings highlight the complexity of the 3CLpro substrate degradome and provide new insights to facilitate the development of anti-CoV drugs.

Cretaceous fossil reveals a new pattern in mammalian middle ear evolution.

The evolution of the mammalian middle ear is thought to provide an example of 'recapitulation'-the theory that the present embryological development of a species reflects its evolutionary history. Accumulating data from both developmental biology and palaeontology have suggested that the transformation of post-dentary jaw elements into cranial ear bones occurred several times in mammals1,2. In addition, well-preserved fossils have revealed transitional stages in the evolution of the mammalian middle ear1,3,4. But questions remain concerning middle-ear evolution, such as how and why the post-dentary unit became completely detached from the dentary bone in different clades of mammaliaforms. Here we report a definitive mammalian middle ear preserved in an eobaatarid multituberculate mammal, with complete post-dentary elements that are well-preserved and detached from the dentary bones. The specimen reveals the transformation of the surangular jaw bone from an independent element into part of the malleus of the middle ear, and the presence of a restricted contact between the columelliform stapes and the flat incus. We propose that the malleus-incus joint is dichotomic in mammaliaforms, with the two bones connecting in either an abutting or an interlocking arrangement, reflecting the evolutionary divergence of the dentary-squamosal joint4. In our phylogenetic analysis, acquisition of the definitive mammalian middle ear in allotherians such as this specimen was independent of that in monotremes and therians. Our findings suggest that the co-evolution of the primary and secondary jaw joints in allotherians was an evolutionary adaptation allowing feeding with unique palinal (longitudinal and backwards) chewing. Thus, the evolution of the allotherian auditory apparatus was probably triggered by the functional requirements of the feeding apparatus.

In-Situ Derived Defective Ru Particles Anchored on Ru-Ni Layered Double Hydroxides for Enhanced Alkaline Hydrogen Evolution.

Developing active, stable, and cost-efficient electrocatalysts to replace platinum for the alkaline hydrogen evolution reaction (HER) is highly desirable yet represents a great challenge. Here, it is reported on a facile one-pot synthesis of RuxNi layered double hydroxides (RuxNi-LDHs) that exhibit remarkable HER activity and stability after an in-situ activation treatment, surpassing most state-of-the-art Ru-based catalysts as well as commercial Ru/C and Pt/C catalysts. The structural and chemical changes triggered by in-situ activation are systematically investigated, and the results clearly show that the pristine, less-active RuxNi-LDHs are transformed into a highly active catalyst characterized by raft-like, defect-rich Ru° particles decorated on the surface of RuxNi-LDHs. Density functional theory (DFT) calculations reveal that the defective Ru sites can effectively optimize the reaction pathway and lower the free energies of the elemental steps involved, leading to enhanced intrinsic activity. This work highlights the importance of the currently understudied strategy of defect engineering in boosting the HER activity of Ru-based catalysts and offers an effective approach involving in-situ electrochemical activation for the development of high-performance alkaline HER catalysts.

Rb-Doped Perovskite Oxides: Surface Enrichment and Structural Reconstruction During the Oxygen Evolution Reaction.

Alkali-metal doped perovskite oxides have emerged as promising materials due to their unique properties and broad applications in various fields, including photovoltaics and catalysis. Understanding the complex interplay between alkali metal doping, structural modifications, and their impact on performance remains a crucial challenge. In this study, this challenge is addressed by investigating the synthesis and properties of Rb-doped perovskite oxides. These results reveal that the doping of Rb into perovskite oxides function as a structural modifier in the as-synthesized samples and during the oxygen evolution reaction (OER) as well. Electron microscopy and first-principles calculations confirm the enrichment of Rb on the surface of the as-synthesized sample. Further investigations into the electrocatalytic reaction revealed that the Rb-doped perovskite underwent drastic restructuring with Rb leaching and formation of strontium oxide.

LEIFR-Net: light estimation for implicit face relight network.

Relighting facial images based on estimated lighting distribution and intensity from image backgrounds and environments can lead to more natural and convincing effects across diverse settings. In this paper, we introduce the Light Estimation for Implicit Face Relight Network (LEIFR-Net), which we believe to be a novel approach that significantly improves upon current methodologies. Initially, we present a method to estimate global illumination from a single image. We then detail our approach for structurally disentangled relighting of faces using pixel-aligned implicit functions. Furthermore, we elaborate on constructing a paired synthetic dataset, which includes environments, maps of lighting distribution, albedo and relighted faces, utilizing a process we refer to as stable diffusion. Our experimental results, evaluated against specific benchmarks, demonstrate the effectiveness of LEIFR-Net in achieving more harmonious alignment of highlights and shadows with environmental lighting, surpassing the performance of other contemporary methods in this domain.

Modeling and optimizing through plenoptic function for the dual lenticular lens-based directional autostereoscopic display system.

We propose an autostereoscopic display system that ensures full resolution for multiple users by directional backlight and eye tracking technology. The steerable beam formed by directional backlight can be regarded as the result of sparsely sampling the light field in space. Therefore, we intuitively propose an optimization algorithm based on the characterization for the state of the steerable beams, which is computed in matrix form using the plenoptic function. This optimization algorithm aims to optimize the exit pupil quality and ultimately enhancing the viewing experience of stereoscopic display. Numerical simulations are conducted and the improvement in exit pupil quality achieved by the optimization scheme is verified. Furthermore, a prototype of the stereoscopic display that employs dual-lenticular lens sheets for the directional backlight has been constructed using the optimized optical parameters. It provides 9 independent exit pupils at the optimal viewing distance of 400 mm, with an exit pupil resolution of 1/30. The field of view is ±16.7°, the viewing distance range is 380 mm to 440 mm. At the optimal viewing distance 400 mm, the average crosstalk of the system is 3%, and the dynamic brightness uniformity across the entire viewing plane reaches 85%. The brightness uniformity of the display at each exit pupil is higher than 88%.

EspalomaCharge: Machine Learning-Enabled Ultrafast Partial Charge Assignment.

Atomic partial charges are crucial parameters in molecular dynamics simulation, dictating the electrostatic contributions to intermolecular energies and thereby the potential energy landscape. Traditionally, the assignment of partial charges has relied on surrogates of ab initio semiempirical quantum chemical methods such as AM1-BCC and is expensive for large systems or large numbers of molecules. We propose a hybrid physical/graph neural network-based approximation to the widely popular AM1-BCC charge model that is orders of magnitude faster while maintaining accuracy comparable to differences in AM1-BCC implementations. Our hybrid approach couples a graph neural network to a streamlined charge equilibration approach in order to predict molecule-specific atomic electronegativity and hardness parameters, followed by analytical determination of optimal charge-equilibrated parameters that preserve total molecular charge. This hybrid approach scales linearly with the number of atoms, enabling for the first time the use of fully consistent charge models for small molecules and biopolymers for the construction of next-generation self-consistent biomolecular force fields. Implemented in the free and open source package EspalomaCharge, this approach provides drop-in replacements for both AmberTools antechamber and the Open Force Field Toolkit charging workflows, in addition to stand-alone charge generation interfaces. Source code is available at https://github.com/choderalab/espaloma-charge.

An in-depth analysis of postoperative insomnia in elderly patients and its implications on rehabilitation.

OBJECTIVES: (1) Assess the prevalence of postoperative insomnia; (2) identify the risk factors for postoperative insomnia before exposure to surgery; (3) explore the impact of postoperative insomnia on rehabilitation. METHODS: A study was conducted with 132 participants aged ≥ 65 undergoing spine interbody fusion. We collected the basic demographic data, Numeric Rating Scales (NRS), Pittsburgh Sleep Quality Index (PSQI), Geriatric Depression Scale (GDS), and Beck Anxiety Inventory (BAI). We measured Quality of Recovery 40 (QoR-40), GDS, BAI, NRS, and PSQI on the first and third nights post-surgery, followed by QoR-40 and NRS assessments two weeks after surgery. RESULTS: The cases of postoperative insomnia on the first and third nights and after two weeks were 81 (61.36%), 72 (54.55%), and 64 (48.48%), respectively, and the type of insomnia was not significantly different (P = 0.138). Sleep efficiency on the first night was 49.96% ± 23.51. On the first night of postoperative insomnia, 54 (66.67%) cases were depression or anxiety, and the PSQI was higher in this group than in the group without anxiety or depression (P < 0.001). PSQI, GDS, and the time of surgery were related factors for postoperative insomnia (PPSQI < 0.001, PGDS = 0.008, and PTime = 0.040). Postoperative rehabilitation showed differences between the insomnia and non-insomnia groups (P < 0.001). CONCLUSIONS: The prevalence of postoperative insomnia in the elderly was high, and postoperative insomnia had a significant correlation with postoperative rehabilitation. Interventions that target risk factors may reduce the prevalence of postoperative insomnia and warrant further research. CLINICAL TRIAL REGISTRATION: Multivariate analysis of postoperative insomnia in elderly patients with spinal surgery and its correlation with postoperative rehabilitation ( https://www.chictr.org.cn/bin/project/edit?pid=170201 ; #ChiCTR2200059827).

Genome-Wide Detection of Copy Number Variations and Their Potential Association with Carcass and Meat Quality Traits in Pingliang Red Cattle.

Copy number variation (CNV) serves as a significant source of genetic diversity in mammals and exerts substantial effects on various complex traits. Pingliang red cattle, an outstanding indigenous resource in China, possess remarkable breeding value attributed to their tender meat and superior marbling quality. However, the genetic mechanisms influencing carcass and meat quality traits in Pingliang red cattle are not well understood. We generated a comprehensive genome-wide CNV map for Pingliang red cattle using the GGP Bovine 100K SNP chip. A total of 755 copy number variable regions (CNVRs) spanning 81.03 Mb were identified, accounting for approximately 3.24% of the bovine autosomal genome. Among these, we discovered 270 potentially breed-specific CNVRs in Pingliang red cattle, including 143 gains, 73 losses, and 54 mixed events. Functional annotation analysis revealed significant associations between these specific CNVRs and important traits such as carcass and meat quality, reproduction, exterior traits, growth traits, and health traits. Additionally, our network and transcriptome analysis highlighted CACNA2D1, CYLD, UBXN2B, TG, NADK, and ITGA9 as promising candidate genes associated with carcass weight and intramuscular fat deposition. The current study presents a genome-wide CNV map in Pingliang red cattle, highlighting breed-specific CNVRs, and transcriptome findings provide valuable insights into the underlying genetic characteristics of Pingliang red cattle. These results offer potential avenues for enhancing meat quality through a targeted breeding program.

Distilling Accurate Descriptors from Multi-Source Experimental Data for Discovering Highly Active Perovskite OER Catalysts.

Perovskite oxides are promising catalysts for the oxygen evolution reaction, yet the huge chemical space remains largely unexplored due to the lack of effective approaches. Here, we report the distilling of accurate descriptors from multi-source experimental data for accelerated catalyst discovery by using the newly designed method of sign-constrained multi-task learning within the framework of sure independence screening and sparsifying operator that overcomes the challenge of data inconsistency between different sources. While many previous descriptors for the catalytic activity were proposed based on respective small data sets, we obtained a new 2D descriptor (dB, nB) based on 13 experimental data sets collected from different publications. Great universality and predictive accuracy, and the bulk-surface correspondence, of this descriptor have been demonstrated. With this descriptor, hundreds of unreported candidate perovskites with activity greater than the benchmark catalyst Ba0.5Sr0.5Co0.8Fe0.2O3 were identified from a large chemical space. Our experimental validations on five candidates confirmed the three highly active perovskite catalysts SrCo0.6Ni0.4O3, Rb0.1Sr0.9Co0.7Fe0.3O3, and Cs0.1Sr0.9Co0.4Fe0.6O3. This work provides an important new approach in dealing with inconsistent multi-source data for applications in the field of data-driven catalysis and beyond.

Mechanisms of polydatin against spinal cord ischemia-reperfusion injury based on network pharmacology, molecular docking and molecular dynamics simulation.

BACKGROUND: Polydatin has shown considerable pharmacological activities in ischemia-reperfusion injuries of various organs. However, its effects and mechanisms in spinal cord ischemia-reperfusion injury have not been fully established. In this study, the mechanisms of polydatin against spinal cord ischemia-reperfusion injury were investigated via network pharmacology, molecular docking and molecular dynamics simulation. METHODS: Spinal cord ischemia-reperfusion injury-related targets were obtained from the GeneCards database, while polydatin-related action targets were obtained from the CTD and SwissTarget databases. A protein-protein interaction network of potential targets was constructed using the String platform. After selecting the potential key targets, GO functional enrichment and KEGG pathway enrichment analyses were performed via the Metascape database, and a network map of "drug-target-pathway-disease" constructed. The relationships between polydatin and various key targets were assessed via molecular docking. Molecular dynamics simulation was conducted for optimal core protein-compound complexes obtained by molecular docking. RESULTS: Topological analysis of the PPI network revealed 14 core targets. GO functional enrichment analysis revealed that 435 biological processes, 12 cell components and 29 molecular functions were enriched while KEGG pathway enrichment analysis revealed 91 enriched signaling pathways. Molecular docking showed that polydatin had the highest binding affinity for MAPK3, suggesting that MAPK3 is a key target of polydatin against spinal cord ischemia-reperfusion injury. Molecular dynamics simulations revealed good binding abilities between polydatin and MAPK3. CONCLUSIONS: Polydatin exerts its effects on spinal cord ischemia-reperfusion injury through multiple targets and pathways. MAPK3 may be a key target of polydatin in spinal cord ischemia-reperfusion injury.

A Jurassic gliding euharamiyidan mammal with an ear of five auditory bones.

Gliding is a distinctive locomotion type that has been identified in only three mammal species from the Mesozoic era. Here we describe another Jurassic glider that belongs to the euharamiyidan mammals and shows hair details on its gliding membrane that are highly similar to those of extant gliding mammals. This species possesses a five-boned auditory apparatus consisting of the stapes, incus, malleus, ectotympanic and surangular, representing, to our knowledge, the earliest known definitive mammalian middle ear. The surangular has not been previously identified in any mammalian middle ear, and the morphology of each auditory bone differs from those of known mammals and their kin. We conclude that gliding locomotion was probably common in euharamiyidans, which lends support to idea that there was a major adaptive radiation of mammals in the mid-Jurassic period. The acquisition of the auditory bones in euharamiyidans was related to the formation of the dentary-squamosal jaw joint, which allows a posterior chewing movement, and must have evolved independently from the middle ear structures of monotremes and therian mammals.

Nonlinear connection between remnant cholesterol and stroke risk: evidence from the China health and retirement longitudinal study.

OBJECTIVE: The evidence on the relationship between remnant cholesterol (RC) and stroke remains controversial. Therefore, this study aimed to explore the relationship between RC and stroke risk in a Chinese population of middle-aged and elderly individuals. METHODS: The present study included 10067 Chinese subjects of middle-aged and elderly individuals. The connection between RC and incident stroke was investigated using the multivariate Cox proportional hazards regression model, several sensitivity analyses, generalized additive models, and smoothed curve fitting. RESULTS: A total of 1180 participants with stroke were recorded during the follow-up period. The multivariate Cox proportional hazards regression model identified a positive connection between RC and stroke risk (hazard ratio (HR) = 1.087, 95% confidence interval (CI): 1.001-1.180). In addition, the current study discovered a nonlinear connection between RC and incident stroke, and the point of inflection for RC was 1.78 mmol/L. The risk of stroke increased by 25.1% with each unit increase in RC level when RC was < 1.78 mmol/L (HR:1.251, 95%CI: 1.089-1.437, P = 0.0015). The results were not affected by sensitivity tests. CONCLUSION: The current study showed a positive and nonlinear connection between RC and stroke risk in a middle-aged and elderly Chinese population. These findings provided new information to help researchers better understand the relationship between RC levels and incident stroke.

Optimization and adsorption characteristics of beads based on heat-inactivated bacterial biomaterial towards the pesticide Cypermethrin.

AIMS: Beads containing heat-inactivated bacterial biomaterial (BBBs) were prepared for removal of cypermethrin (CPM) and the conditions for this removal were evaluated and optimized via orthogonal experiments. The adsorption characteristics of BBBs and the binding mechanism were then explored. METHODS AND RESULTS: Single-factor and orthogonal experiments were carried out to optimize five factors affecting the production and effectivity of the beads. The adsorption rate of CPM could reach 98% with beads prepared under optimized conditions: equal volumes of Lactobacillus cell debris derived from 1 × 1011 CFU; 2% hydroxypropyl-ß-cyclodextrin and 2.5% activated carbon concentration, were mixed to give mixture TM, and this and SA, was mixed 1:4 with sodium alginate (SA) and beads were prepared using a 26-Gauge needle). The best adsorption conditions were initial CPM concentration of 10 mg l-1, incubation time of 24 h, and rotational speed of 180 rpm. BBBs have a well-formed structure and abundant surface functional groups, such as -COOH, -OH, -NH, -CH, -CO, -C = C. The adsorption process conformed to pseudo-second-order kinetic, and it was also a Freundlich monolayer adsorption, and the calculated maximum adsorption capacity was 9.69 mg g-1 under optimized conditions. CONCLUSIONS: BBBs showed the highest CPM removal capacity and a good tolerance ability.

Optimization and adsorption characteristics of beads based on heat-inactivated bacterial biomaterial towards the pesticide Cypermethrin.

AIMS: Beads containing heat-inactivated bacterial biomaterial (BBBs) were prepared for removal of cypermethrin (CPM) and the conditions for this removal were evaluated and optimized via single-factor coupled orthogonal experiments based on five factors. The adsorption characteristics of BBBs and the binding mechanism were then explored. METHODS AND RESULTS: Results showed that the adsorption rate of CPM could reach 98% with beads prepared under optimized conditions: equal volumes of Lactobacillus cell debris derived from 1×1011 CFU; 2% hydroxypropyl-ß-cyclodextrin and 2.5% activated carbon concentration, were mixed to give mixture TM, and this and SA, was mixed 1:4 with sodium alginate (SA) and beads were prepared using a 26-Gauge needle). The best adsorption conditions were initial CPM concentration of 10 mg l-1, incubation time of 24 h, and rotational speed of 180 rpm. BBBs have a well-formed structure and abundant surface functional groups, such as -COOH, -OH, -NH, -CH, -CO, -C=C. The adsorption process conformed to pseudo-second-order kinetic, and it was also a Freundlich monolayer adsorption, and the calculated maximum adsorption capacity was 9.69 mg g-1 under optimized conditions. CONCLUSIONS: BBBs showed the highest CPM removal capacity and a good tolerance ability. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results provided a theoretical foundation for developing an adsorbent with heat-inactivated Lactobacillus plantarum (L. plantarum) RS60 for removing CPM in wastewater or drinks.

Remote Sensing of Seawater Temperature and Salinity Profiles by the Brillouin Lidar Based on a Fizeau Interferometer and Multichannel Photomultiplier Tube.

Brillouin spectroscopy is a powerful tool to measure the water temperature and salinity profiles of seawater. Considering the insufficiency of the current spectral measurement methods in real-time, spectral integrity, continuity, and stability, we developed a new lidar system for spectrum measurement on an airborne platform that is based on a Fizeau interferometer and multichannel photomultiplier tube. In this approach, the lidar system uses time-of-flight information to measure the depth and relies on Brillouin spectroscopy as the temperature and salinity indicator. In this study, the system parameters were first optimized and analyzed. Based on the analysis results, the performance of the system in terms of detection depth and accuracy was evaluated. The results showed that this method has strong anti-interference ability, and under a temperature measurement accuracy of 0.5 °C and a salinity measurement accuracy of 1‱, the effective detection depth exceeds 40.51 m. Therefore, the proposed method performs well and will be a good choice for achieving Brillouin lidar application in seawater remote sensing.

Wavelength-multiplexed stereoscopic LC display using scanning backlight.

A stereoscopic display system based on a wavelength-multiplexed and time-multiplexed technique is introduced. The system includes white LEDs, complementary multiband bandpass filters, 30 double-side-lit light guide bars, and a liquid crystal display panel, as well as a pair of complementary multiband bandpass filter glasses. The LEDs are divided into two groups, and each group is covered with a kind of complementary multiband bandpass filter. They can be controlled by the driving circuit and work in the scanning mode in synchronization with the liquid crystal display panel, as the liquid crystal display panel displaying the left and right images frame by frame. The backlight for the left and right images is sampled by complementary multiband bandpass filter1 and complementary multiband bandpass filter2, respectively, and can only pass through the corresponding glass lens, realizing channel separation. A prototype based on this principle was set up and experiments were carried out to evaluate the performance of the system. Double-side-lit backlight with light guide bars ensures the uniformity of the backlight while using fewer LEDs, and therefore reducing power consumption. Brightness through complementary multiband bandpass filter1 and complementary multiband bandpass filter2 is about 50.5 nits and 55.5 nits, respectively. It is demonstrated in our system that the scanning backlight with 30 light guide bars is capable of reducing cross talk to 2.2%. Furthermore, the resolution in the wavelength-multiplexed 3D mode is the same as the physical resolution of the liquid crystal display panel.

A comparative study on auditory and hyoid bones of Jurassic euharamiyidans and contrasting evidence for mammalian middle ear evolution.

The holotypes of euharamiyidan Arboroharamiya allinhopsoni and Arboroharamiya jenkinsi preserve the auditory and hyoid bones, respectively. With additional structures revealed by micro-computerized tomography (CT) and X-ray micro-computed laminography (CL), we provide a detailed description of these minuscule bones. The stapes in the two species of Arboroharamiya are similar in having a strong process for insertion of the stapedius muscle. The incus is similar in having an almond-shaped body and a slim short process, in addition to a robust stapedial process with a short lenticular process preserved in A. allinhopsoni. The plate-like ectotympanic in the two species of Arboroharamiya is similar and comparable to that of Qishou jizantang. The surangular in the two species has a fan-shaped body and a needle-shaped anterior process. The malleus, ectotympanic, and surangular are fully detached from the dentary and should have functioned exclusively for hearing. All the auditory bones of Arboroharamiya display unique features unknown in other mammaliaforms. Moreover, hyoid elements are found in the two species of Arboroharamiya and co-exist with the five auditory bones in the holotype of A. allinhopsoni. The element interpreted as the stylohyal is similar to the bone identified as the ectotympanic in Vilevolodon. We reconstruct the auditory apparatus of Arboroharamiya and compare it with that of Vilevolodon as well as those in extant mammals and basal mammaliaforms. The comparison shows diverse morphological patterns of the auditory region in mammaliaforms. In particular, those of Vilevolodon and Arboroharamiya differ significantly: the former has a mandibular middle ear, whereas the latter possesses a definitive mammalian middle ear. It is puzzling that the two sympatric and dentally similar taxa have such different auditory apparatuses. In light of the available evidence, we argue that the mandibular middle ear reconstructed in Vilevolodon encounters many problems, and the so-called ectotympanic in Vilevolodon may be interpreted as a stylohyal; thus, the dilemma can be resolved.

Three new Jurassic euharamiyidan species reinforce early divergence of mammals.

The phylogeny of Allotheria, including Multituberculata and Haramiyida, remains unsolved and has generated contentious views on the origin and earliest evolution of mammals. Here we report three new species of a new clade, Euharamiyida, based on six well-preserved fossils from the Jurassic period of China. These fossils reveal many craniodental and postcranial features of euharamiyidans and clarify several ambiguous structures that are currently the topic of debate. Our phylogenetic analyses recognize Euharamiyida as the sister group of Multituberculata, and place Allotheria within the Mammalia. The phylogeny suggests that allotherian mammals evolved from a Late Triassic (approximately 208 million years ago) Haramiyavia-like ancestor and diversified into euharamiyidans and multituberculates with a cosmopolitan distribution, implying homologous acquisition of many craniodental and postcranial features in the two groups. Our findings also favour a Late Triassic origin of mammals in Laurasia and two independent detachment events of the middle ear bones during mammalian evolution.

Quantitative analysis of multicomponents by single marker combined with HPLC fingerprint qualitative analyses for comprehensive evaluation of Aurantii Fructus.

The present study aimed to develop a strategy involving quantitative analysis of multicomponents by single marker in combination with high-performance liquid chromatography fingerprint qualitative analysis for performing the quality control of Aurantii Fructus. The content of 12 components (eriocitrin, neoeriocitrin, narirutin, naringin, hesperidin, neohesperidin, meranzin, poncirin, naringenin, nobiletin, tangeretin, and auraptene) in samples was determined using reliable relative correction factors that were obtained using naringin as an internal reference standard. The new method demonstrated good applicability, and no significant differences were observed between the external standard method and the new method as determined by calculating standard method difference. Qualitative evaluation of samples was conducted using similarity analysis, hierarchical cluster analysis, and quality fluctuation analysis. Chromatographic fingerprint data were divided into three groups by similarity and hierarchical cluster analyses, and seven components may have a more significant impact on the quality of Aurantii Fructus in quality fluctuation analysis. Overall, the study suggests that the qualitative and quantitative analyses of multicomponents using quantitative analysis of multicomponents by single marker combined with chromatographic fingerprinting can be considered good quality criteria for performing quality control and providing technical support for the further pharmacological and pharmaceutical research of Aurantii Fructus.