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Seasonal variation has been shown to influence symbiotic bacterial community composition and structure in amphibians. It is still unknown how the symbiotic bacterial community assembly changes during different seasons, especially for amphibians who are particularly sensitive to environmental change. We found significant differences in the composition and diversity (alpha and beta diversity) of amphibian skin and gut bacteria. Co-occurrences network analysis showed that seasonal variation reduced the microbial network complexity of amphibians from summer to autumn. The normalized stochastic ratio (NST) and phylogenetic bin-based null model analysis (iCAMP) models showed that the same result that stochastic processes was the major factor regulating the symbiotic bacterial community assembly mechanisms of amphibians. From summer to autumn, the symbiotic bacterial community assembly mechanisms declined in the contribution of stochastic processes, while increasing in the contribution of deterministic processes. Dispersal limitation was the dominant microbial assembly mechanism, followed by homogeneous selection, and then heterogeneous selection in the symbiotic bacterial community communities of amphibians between summer and autumn. Furthermore, higher niche width of the symbiotic bacterial community of amphibians was found in summer than autumn. Overall, these results demonstrated that seasonal variation influenced amphibian symbiotic bacterial community between summer and autumn.

The studies of climatic‐niche shifts over evolutionary time accompanied by key morphological innovations have attracted the interest of many researchers recently. We applied ecological niche models (ENMs), ordination method (environment principal component analyses; PCA‐env), combined phylogenetic comparative methods (PCMs), and phylogenetic generalized least squares (PGLS) regression methods to analyze the realized niche dynamics and correspondingly key morphological innovations across clades within Scutiger boulengeri throughout their distributions in Qinghai–Tibet Plateau (QTP) margins of China. Our results show there are six clades in S. boulengeri and obvious niche divergences caused by niche expansion in three clades. Moreover, in our system, niche expansion is more popular than niche unfilling into novel environmental conditions. Annual mean temperature, annual precipitation, and precipitation of driest month may contribute to such a shift. In addition, we identified several key climatic factors and morphological traits that tend to be associated with niche expansion in S. boulengeri clades correspondingly. We found phenotypic plasticity [i.e., length of lower arm and hand (LAHL), hind‐limb length (HLL), and foot length (FL)] and evolutionary changes [i.e., snout–vent length (SVL)] may together contribute to niche expansion toward adapting novel niche, which provides us a potential pattern of how a colonizing toad might seed a novel habitat to begin the process of speciation and finally adaptive radiation. For these reasons, persistent phylogeographic divisions and accompanying divergences in niche occupancy and morphological adaption suggest that for future studies, distinct genetic structure and morphological changes corresponding to each genetic clade should be included in modeling niche evolution dynamics, but not just constructed at the species level., Persistent phylogeographic divisions and accompanying divergences in niche occupancy and morphological adaption suggest that for future studies, distinct genetic structure and morphological changes corresponding to each genetic clade should be included in modeling niche evolution dynamics, but not just constructed at the species

Describing osteological development is of great importance for understanding vertebrate phenotypic variations, form-functional transitions and ecological adaptations. Anurans exhibit dramatic changes in their morphology, habitat preferences, diet and behaviour between the tadpole and frog stages. However, the anatomical details of their cranial and postcranial development have not been extensively studied, especially in Microhylidae. In this work, we studied the microhylid Microhyla fissipes, commonly known as the ornamented pygmy frog, a small-sized frog with fast metamorphosis. Its osteological development was comprehensively described based on 120 cleared and stained specimens, including six tadpoles for each stage between 28 and 45, six juveniles and six adults. Additionally, 22 osteological traits of these specimens involved in food acquisition, respiration, audition and locomotion were selected and measured to reflect the changes in tadpole ecological functions during metamorphosis. Our study provides the first detailed qualitative and quantitative developmental information about these structures. Our results have confirmed that skeletal elements (viz., neopalatines, omosternum, clavicles and procoracoids) absent in adults are not detected during development. Our data reveal that morphologically, radical transformations of the cranial structures related to feeding and breathing are completed within stages 42-45 (72 h), but the relative length and width of these skeletons have changed in earlier stages. The postcranial skeletons correlated with locomotion are well developed before stage 42 and approach the adult morphology at stage 45. Indeed, the relative length of the pectoral girdle and forelimb reaches the adult level at stage 42 and stage 45, respectively, whereas that of the vertebral column, pelvic girdle and hind limbs increases from their appearance until reaching adulthood. Based on published accounts of 19 species from Neobatrachia, Mesobatrachia and Archaeobatrachia, cranial elements are among the first ossified skeletons in most studied species, whereas sphenethmoids, neopalatines, quadratojugals, mentomeckelians, carpals and tarsals tend to ossify after metamorphosis. These results will help us to better understand the ecomorphological transformations of anurans from aquatic to terrestrial life. Meanwhile, detailed morphological and quantitative accounts of the osteological development of Microhyla fissipes will provide a foundation for further study.

We consider the Ising model at its critical temperature with external magnetic field $ha^{15/8}$ on the square lattice with lattice spacing $a$. We show that the truncated two-point function in this model decays exponentially with a rate independent of $a$. As a consequence, we show exponential decay in the near-critical scaling limit Euclidean magnetization field. For the lattice model with $a=1$, the mass (inverse correlation length) is of order $h^{8/15}$ as $h\downarrow 0$; for the Euclidean field, it equals exactly $Ch^{8/15}$ for some $C$. Although there has been much progress in the study of critical scaling limits, results on near-critical models are far fewer due to the lack of conformal invariance away from the critical point. Our arguments combine lattice and continuum FK representations, including coupled conformal loop and measure ensembles, showing that such ensembles can be useful even in the study of near-critical scaling limits. Thus we provide the first substantial application of measure ensembles.

Ecologists have generally concluded that species distributions are not random (e.g. aggregate), based on single-species studies that applied single-species-based statistical methods, like the negative binomial model. Although it is common knowledge that some specific species in an ecological community present aggregate distributions, this does not necessarily imply that the entire community presents an aggregate distribution. Studying community-level distributional aggregation patterns requires different statistical methods. Herein, by utilizing a novel conspecific-encounter index derived from a multiple species Markov transition model that accounts for the non-independent sampling of consecutive individuals along line transects, we were able to show that tree assemblages in tropical forest ecosystems can present a strong signal of extensive distributional interspersion. This interesting result is not unexpected, given the fact that neighbouring tree individuals in highly diverse tropical forests are usually of different species, resulting in strong niche packing or interspersed patterns. In contrast, for the amphibian assemblages surveyed from southwestern China and central-south Vietnam, the conspecific-encounter index was found to be consistently high, implying that amphibian communities tend to be highly aggregate in space. Conclusively, using the conspecific-encounter index derived from the Markov non-independent sampling model, we provide a legible definition of community-level distributional aggregation as an interspersed or cluster-like distribution of different species. This definition is not idiosyncratic, as it is coincident with the definition of the contagion index used in landscape ecology. To this end, the model used in this paper establishes a framework explicitly linking community ecology and landscape ecology from a multi-object perspective.

Mountains, representing storehouses of biodiversity, endemism and threatened species, are biodiversity hotspots of great conservation importance. However, increasing evidence indicates that mountain species throughout the world are responding to climate change, past or contemporary, by shifting their geographic distributions and patterns of genetic diversity, potentially affecting their adaptive capacity and increasing risk of extinction. Using the iconic high-elevation frog Nanorana parkeri as indicator, we showed how spatial analyses of climatic stability combined with genetic data allow unravelling amphibian responses to past and future climate changes on 'the roof of the world'-the Qinghai-Tibetan Plateau. We found that areas along the Yarlung Tsangpo Valley were climatically more stable relative to other regions, apparently serving as a large climatic refugium during Quaternary glaciations, but that these areas will likely be affected by future climate change. As populations closer to Quaternary refugia usually had higher genetic diversity, current genetic diversity can be explained in the largest part by distance to historically stable areas, outweighing other historical and contemporary factors. Along with the dynamics of suitable range, a fluctuating habitat fragmentation supported the pattern of historical changes in genetic diversity (Ne ) over time. Our results emphasize strong relationships between amphibian genetic diversity, past range dynamics and where to preserve suitable habitats in the face of future climate changes. More generally, our findings highlighted a central role of refugia during Quaternary climatic fluctuations, and how isolation from refugia may have modulated amphibian genetic diversity across the Qinghai-Tibetan Plateau.

A novel kind of coronavirus has infected more than 1, 600, 000 people, claimed over 100, 000 lives, and spread to 212 countries and territories since December 2019.1 The virus has been named SARS‐CoV‐2 (provisionally termed 2019‐nCoV before2), and the pneumonia caused by it is COVID‐19. Though many scientists and researchers are trying to identify the animals harboring SARS‐CoV‐2, the intermediate host remains under the veil.This article is protected by copyright. All rights reserved.

Protected areas are considered as an essential strategy to halt the decline of biodiversity. Ecological representation in protected areas is crucial for assessment on the progress toward conservation targets. Although China has established a large number of protected areas since the 1950s, ecological representation of protected areas is poorly understood. Here, we performed the complementarity analysis to evaluate ecological representation of protected areas in China. We used a database of the geographical distribution for 10,396 woody plant species, 2,305 fern species, 406 amphibian species, 460 reptile species, 1,364 bird species, and 590 mammal species from 2,376 counties across China. We identified complementary sets of counties for all species or threatened species of plant and vertebrate species using a complementarity algorithm. We evaluated ecological representation of 3,627 protected areas and discerned conservation gaps by comparing the distribution of protected areas with complementary sets. The results show that the spatially representative and complementary sites for biodiversity are poorly covered, and a fairly large proportion of protected areas is not designed to efficiently represent biodiversity at the national scale. Our methodology can serve as a generic framework for assessment on ecological representation of protected areas at the national scale.

Functional traits are increasingly recognized as an integrative approach by ecologists to quantify a key facet of biodiversity. And these traits are primarily expressed as species means in previous studies, based on the assumption that the effects of intraspecific variability can be overridden by interspecific variability when studying functional ecology at the community level. However, given that intraspecific variability could also have important effects on community dynamics and ecosystem functioning, empirical studies are needed to investigate the importance of intraspecific variability in functional traits. In this study, 256 Scutiger boulengeri tadpole individuals from four different populations are used to quantify the functional difference between populations within a species, and the relative contribution of inter‐ and intrapopulation variability in functional traits. Our results demonstrate that these four populations differ significantly in functional attributes (i.e., functional position, functional richness, and low functional overlap), indicating that individuals from different populations within a species should be explicitly accounted for in functional studies. We also find similar relative contribution of inter‐ (~56%) and intrapopulation (~44%) variation to the total variability between individuals, providing evidence that individuals within populations should also be incorporated in functional studies. Overall, our results support the recent claims that intraspecific variability cannot be ignored, as well as the general idea of “individual level” research in functional ecology.

Background Next‐generation sequencing (NGS) has been widely used in both clinics and research. It has become the most powerful tool for diagnosing genetic disorders and investigating disease etiology through the discovery of genetic variants. Variants identified by NGS are stored in variant call format (VCF) files. However, querying and filtering VCF files are extremely difficult for researchers without programming skills. Furthermore, as the mutation data are increasing exponentially, there is an urgent need to develop tools to manage these variant data in a centralized way. Methods The VCF‐Server was developed as a web‐based visualization tool to support the interactive analysis of genetic variant data. It allows researchers and medical geneticists to manage, annotate, filter, query, and export variants in a fast and effective way. Results In this study, we developed the VCF‐Server, a powerful and easily accessible tool for researchers and medical geneticists to perform variant analysis. Users can query VCFs, annotate, and filter variants without knowing programming code. Once the VCF file is uploaded, VCF‐Server allows users to annotate the VCF with commonly used databases or user‐defined variant annotations (including variant blacklist and whitelist). Variant information in the VCF is shown visually via the interactive graphical interface. Users can filter the variants with flexible filtering rules, and the prioritized variants can be exported locally for further analysis. As VCF‐Server adopts a web file system, files in the VCF‐Server can be stored and managed in a centralized way. Moreover, VCF‐Server allows direct web‐based analysis (accessible through either desktop computers or mobile devices) as well as local deployment. Conclusions With an easy‐to‐use graphical interface, VCF‐Server allows researchers with little bioinformatics background to explore and mine mutation data, which may broaden the application of NGS technology in clinics and research. The tool is freely available for use at https://www.diseasegps.org/VCF-Server?lan = eng.

Aim Protected areas are an important tool for conserving species. In this study, we assessed the effectiveness of protected areas to conserve amphibian biodiversity in response to future changes in climate and land use. Location China. Methods Range maps and occurrence records of amphibian species in China were analysed separately using ensemble species distribution modelling across three spatial scales to assess scale dependency. Climate velocity and corresponding residence time in protected areas and species’ ranges were calculated, together with a number of other effectiveness indices. Results Predicted declines in amphibian richness, endemism, phylogenetic diversity, phylogenetic endemism and suitable habitat were lower in protected than in unprotected areas, complementary-priority sites or richness hotspots. However, less-disturbed amphibian habitat, calculated from current and future projected land use data, in both protected and unprotected areas were consistently lost over time although this reduction was lower in protected areas. Although residence time of precipitation was longer in protected areas and within species’ ranges in protected areas, resident time of temperature was significantly shorter in both. These results were consistent regardless of data sources and spatial scales. Main conclusions China's current protected areas are predicted to maintain future amphibian distribution and diversity, but are insufficient in preventing the losses of suitable climate and areas of less-disturbed habitat. The top 10% of future conservation gaps for amphibians were identified in China based on performance of effectiveness indices. The two largest gaps prioritized for future protected areas include the southern parts of Tibet and the Hengduan Mountains.

Species of the genus Gynandropaa within the family Dicroglossidae are typical spiny frogs whose taxonomic status has long been in doubt. We used integrative methods, involving morphological and molecular analyses, to elucidate the phylogenetic relationships, and to determine identities and the geographic distribution of each valid species. We obtained DNA sequence data of 5 species of Gynandropaa (complete sequences of the mitochondrial NADH dehydrogenase subunit 2 [ND2] gene, and 890 bp of 12S rRNA and 16S rRNA partial sequences) from 37 localities (including the topotypes of 5 described species) and constructed Bayesian and maximum-likelihood trees to examine the patterns of phylogeography. A total of 28 morphological variables were taken on 624 specimens. Three clades with clear geographic patterns were recognized: clade C (from south-western Sichuan Province and central Yunnan Province), clade E (western Guizhou Province and eastern to central Yunnan Province) and clade W (western to southern Yunnan Province). Integrating morphological characteristics and distribution information, the clades W, E and C represent Gynandropaa yunnanensis, G. phrynoides and G. sichuanensis, respectively. We draw the following conclusions: (i) the taxon G. phrynoides, formerly evaluated as a junior synonym of G. yunnanensis, is revalidated herein at the rank of species; (ii) G. liui is a junior synonym of G. sichuanensis; and (iii) G. yunnanensis is a valid species while G. bourreti is probably a subspecies of G. yunnanensis, with the distribution range from Vietnam to southern Yunnan Province. This study clears up the taxonomic status of Gynandropaa and provides important information for understanding the evolution and conservation of these spiny frogs.

In the present study, comprehensive 2D GC-TOF-MS combined with multivariate data analysis was applied to analyze the differences of the volatile components in crude and processed Atractylodis Macrocephalae Rhizoma (AMR) samples. As a result, 26 compounds that were found in crude AMR samples disappeared in processed AMR samples, and 19 compounds were newly generated and identified in AMR after processing with wheat bran. Meanwhile, principal component analysis demonstrated that there were significant chemical differences between crude and processed AMR samples, and processing procedure caused obvious quantitative and qualitative changes of volatile components in AMR. The established method could be used to explain the chemical differentiation between crude and processed AMR, and to further understand the processing mechanism of herbal medicines.

Background and Purpose Mas oncogene-related gene (Mrg) receptors are exclusively distributed in small-sized neurons in trigeminal and dorsal root ganglia (DRG). We investigated the effects of MrgC receptor activation on inflammatory hyperalgesia and its mechanisms. Experimental Approach A selective MrgC receptor agonist, bovine adrenal medulla peptide 8-22 (BAM8-22) or melanocyte-stimulating hormone (MSH) or the μ-opioid receptor (MOR) antagonist CTAP was administered intrathecally (i.t.) in rats injected with complete Freund's adjuvant (CFA) in one hindpaw. Thermal and mechanical nociceptive responses were assessed. Neurochemicals were measured by immunocytochemistry, Western blot, ELISA and RT-PCR. Key Results CFA injection increased mRNA for MrgC receptors in lumbar DRG. BAM8-22 or MSH, given i.t., generated instant short and delayed long-lasting attenuations of CFA-induced thermal hyperalgesia, but not mechanical allodynia. These effects were associated with decreased up-regulation of neuronal NOS (nNOS), CGRP and c-Fos expression in the spinal dorsal horn and/or DRG. However, i.t. administration of CTAP blocked the induction by BAM8-22 of delayed anti-hyperalgesia and inhibition of nNOS and CGRP expression in DRG. BAM8-22 also increased mRNA for MORs and pro-opiomelanocortin, along with β-endorphin content in the lumbar spinal cord and/or DRG. MrgC receptors and nNOS were co-localized in DRG neurons. Conclusions and Implications Activation of MrgC receptors suppressed up-regulation of pronociceptive mediators and consequently inhibited inflammatory pain, because of the activation of up-regulated MrgC receptors and subsequent endogenous activity at MORs. The uniquely distributed MrgC receptors could be a novel target for relieving inflammatory pain.

Aim: Whether the burden of advanced oxidation protein products (AOPP) accumulation, a marker of oxidative stress, is affected by dialysis modality remains unclear. We compared the serum levels of AOPP in patients on haemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) and tested the hypothesis that an accumulation of AOPP was an independent risk factor for cardiovascular disease. Methods: This was a cross-section study. A total of 2095 patients (1539 HD, 556 CAPD) were recruited from the nine largest dialysis centres in China. Persons in medical centres for disease screening were selected as controls. Patients maintained on HD were dialyzed twice or thrice weekly. CAPD patients used lactate-buffered, glucose-containing solutions. The patients' data were abstracted from the medical record. The serum levels of AOPP were determined by spectrophotometric detection. Results: The levels of AOPP were significantly elevated in both HD and CAPD patients compared to healthy controls. Accumulation of AOPP was more significant in HD compared to CAPD population. Meanwhile, AOPP accumulation was associated with the presence of ischaemic heart disease (IHD) only in HD, but not CAPD patients. A higher proportion of IHD was found in the HD population among those with higher levels of AOPP in each category of age and irrespective of the presence or absence of high triglyceride. Multivariate regression analysis indicated that accumulation of AOPP was an independent risk factor for IHD in HD population. Conclusion: Accumulation of AOPP was more significant in HD compared to CAPD patients. The level of AOPP was independently associated with IHD only in HD patients.

Bovine adrenal medulla 22 (BAM22) peptide is one of the cleavage products of proenkephalin A. It binds with high affinity to both opioid receptors and a newly discovered receptor in vitro. This latter receptor was first named sensory neuron-specific receptor and is here named BAM peptide-activated receptor with non-opioid activity (BPAR). BPAR is uniquely distributed in small-diameter DRG neurons, most of which are associated with the IB4 class of nociceptor afferent. The present study examined the effects of intrathecal administration of BAM22 on formalin-induced nocifensive behaviors and tail-withdrawal latency in the rat. Intrathecal (i.t.) administration of BAM22 decreased nocifensive behavior scores, measured as the sum of flinching and lifting/licking, in the first and second phases of the formalin test. This decrease was partially attenuated by systemic injection of naloxone. In the presence of naloxone, i.t. BAM22 produced a dose-dependent suppression of the nocifensive behaviors observed during the formalin test. The ratio of the efficacy of BAM22 (5 nmol) in the presence of naloxone over that in the absence of naloxone was 0.65 for flinching and 0.74 for lifting/licking in the second phase. BAM22 at a dose of 5 nmol increased the tail-withdrawal latency by 193 and 119% of baseline in the absence and presence of naloxone, respectively. Systemic administration of naloxone alone enhanced the nocifensive behaviors in the second, but not in the first phase of the formalin test. Naloxone treatment did not alter the tail-withdrawal latency. These data confirm earlier in vitro data showing that BAM22 has both opioid and non-opioid biological actions. The non-opioid action of BAM22 involves inhibition of acute and persistent nociceptive behaviors at the spinal level, presumably mediated via BPAR. The name suggested for this novel receptor, its potential physiological function and its ligand are discussed. British Journal of Pharmacology (2004) 141, 423–430. doi:10.1038/sj.bjp.0705637

A novel microscopy is developed for evaluating the fracture toughness of thin ceramic sheets using the micro-indentation method. This microscopy overcomes the theoretical limit of measurement resolution associated with optical microscopes. Using a modified optical microscope, the crack lengths produced by micro-indentation on the surface of an as-fired ceramic sheet can now be measured. Experimental results on an 8 mol% yttria-stabilized zirconia ceramic substrate show that the accuracy of the fracture toughness evaluation is satisfactory. This micro-indentation fracture toughness evaluation method is efficient and economical for thin ceramic substrate surfaces, either polished or as-fired.