[Determination of 11 nutrients in liquid milk by ultra-performance liquid chromatography-tandem mass spectrometry].

OBJECTIVE: To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method for simultaneous determination of 11 nutritional components(thiamine, riboflavin, nicotinamide, nicotinic acid, pantothenic acid, pyridoxine, pyridoxal, pyridoxamine, biotin, choline, L-carnitine) in liquid milk. METHODS: Milk samples were shaken with 20 mmol/L ammonium formate solution and heated in a water bath at 100 ℃ for 30 min, then incubated with papain and acid phosphatase at 45 ℃ for 16 h, the lower liquid was collected after centrifugation for analysis. UPLC separation was performed on an ACQUITY~(TM) HSS T3(3.0 mm×150 mm, 1.8 µm) column, 2 mmol/L ammonium formate(containing 0.1% formic acid) solution and acetonitrile(containing 0.1% formic acid) were used as mobile phase. Quantitative detection was performed by internal standard method. RESULTS: 11 nutritional components can be effectively separated and detected in 12 min, and the linear correlation coefficients(R~2) were all above 0.995. The limits of detection(LODs) were between 0.05 and 0.50 µg/L, and the limits of quantification(LOQs) were between 0.20 and 1.25 µg/L. The recovery rates of three-level addition were 85.6%-119.3%, and the precision RSDs were between 3.68% and 7.82%(n=6). Based on the detection of 60 liquid milk samples from 5 different animals, it was found that the contents of 11 nutrients in liquid milk from different milk sources were significantly different, but pyridoxine could not be detected. CONCLUSION: The method can quantitatively detect 11 water-soluble nutrients, including free and bound forms, by effective enzymolysis. It is sensitive, reproducible and can meet the needs of quantitative detection.

Rewiring cis-2-butene-1,4-dial mediated urinary metabolomics fingerprints of short-term exposure to furan.

Furan represents one of the dietary-sourced persistent organic pollutants and thermal processing contaminants. Given its widespread occurrence in food and various toxicological effects, accurately assessing furan exposure is essential for informing public health risks. Furan is metabolized to a reactive primary product, cis-2-butene-1,4-dial (BDA) upon absorption. Some of the resulting BDA-derived metabolites have been proposed as potential exposure biomarkers of furan. However, the lack of quantification for recognized and feasible furan biomarkers has hampered the development of internal exposure risk assessment of furan. In this study, we employed reliable non-targeted metabolomics techniques to uncover urinary furan metabolites and elucidate their chemical structures. We characterized 8 reported and 11 new furan metabolites derived from the binding of BDA with glutathione (GSH), biogenic amines, and/or amino acids in the urine of male rats subjected to varying doses of furan. Notably, a mono-GSH-BDA adduct named cyclic GSH-BDA emerged as a highly prospective specific biomarker of furan exposure, as determined by an ultrahigh-performance liquid chromatography-tandem mass spectrometry method. Cyclic GSH-BDA demonstrated a robust mass spectrometry ion response intensity and exhibited evident time- and dose response. Additionally, we conducted a comprehensive profiling of the kinetics of potential furan biomarkers over time to capture the metabolic dynamics of furan in vivo. Most urinary furan metabolites reached peak concentrations at either the first (3 h) or second (6 h) sampling time point and were largely eliminated within 36 h following furan treatment. The present study provides novel insights into furan metabolism and sheds light on the biomonitoring of furan exposure.

Association between parents' concerns about eating and sleeping problems and social-emotional development in Chinese children aged 3 to 6 years.

Background: Parents' parenting beliefs have a major influence on their children's eating and sleeping problems and emotional socialization. However, the relationship between parent's concerns about eating or sleeping problems and social-emotional development is unclear. Methods: We used a convenience sampling method to investigate 997 parents of preschool children aged 3 to 6 in Hangzhou, China, and asked them to complete the "Ages & Stages Questionnaire: Social-Emotional (2nd Edition)" (ASQ: SE-2) and the Survey of Concerns about Children's Eating and Sleeping Problems. To examine the relationship between children's social-emotional development and their parents' concerns about their eating or sleeping problems, binary logistic regression was used. Results: There were 218 children (21.9%) with a suspected social-emotional development delay, and 273 parents (27.4%) were concerned about their children's eating or sleeping problems, which mainly focused on ill-balanced eating, bad eating habits, and difficulty falling asleep. The rate of suspected social-emotional development delay in children with the co-occurrence of eating and sleeping problems (37.8%) was significantly higher than those with only eating problems (29.7%), only sleeping problems (24.4%), and those with no eating or sleeping problems (18.8%) (p < 0.05). A binary logistic regression analysis showed that parents' concerns about the co-occurrence of eating and sleeping problems (OR = 2.52, p = 0.01) and only eating problems (OR = 1.71, p = 0.004) were risk factors for children's social-emotional development. In addition, boys were more likely than girls to have suspected social-emotional development delay (OR = 1.49, p = 0.01). Conclusion: Children whose parents were concerned about only eating or the co-occurrence of eating and sleeping problems were linked to have a higher risk of suspected social-emotional development delay.

Molecular phylogeny and taxonomy of the genus Vernaya (Mammalia: Rodentia: Muridae) with the description of two new species.

The climbing mouse is a rare, small mammal listed as an endangered species on the China species red list. Molecular phylogenetic analyses and the evolutionary history of the genus remain unexplored because of the extreme difficulty in capturing individuals and their narrow distribution. Here, we collected 44 specimens, sequenced one mitochondrial and eight nuclear genes, and integrated morphological approaches to estimate phylogenetic relationships, delimit species boundaries, and explore evolutionary history. Molecular analyses and morphological results supported the validity of these four species. Here, we describe two new species, Vernaya meiguites sp. nov. and Vernaya nushanensis sp. nov., and recognize Vernaya foramena, previously considered a subspecies of Vernaya fulva, as a valid species. The estimated divergence time suggests that the climbing mouse began to diversify during the Pliocene (3.36 Ma).

Adversarial Attack and Defense in Breast Cancer Deep Learning Systems.

Deep-learning-assisted medical diagnosis has brought revolutionary innovations to medicine. Breast cancer is a great threat to women's health, and deep-learning-assisted diagnosis of breast cancer pathology images can save manpower and improve diagnostic accuracy. However, researchers have found that deep learning systems based on natural images are vulnerable to attacks that can lead to errors in recognition and classification, raising security concerns about deep systems based on medical images. We used the adversarial attack algorithm FGSM to reveal that breast cancer deep learning systems are vulnerable to attacks and thus misclassify breast cancer pathology images. To address this problem, we built a deep learning system for breast cancer pathology image recognition with better defense performance. Accurate diagnosis of medical images is related to the health status of patients. Therefore, it is very important and meaningful to improve the security and reliability of medical deep learning systems before they are actually deployed.

Molecular phylogenetics and diversity of the Himalayan shrew ( Soriculus nigrescens Gray, 1842) (Eulipotyphla, Soricidae) in Southwest China.

The Himalayan shrew, Soriculus nigrescens Gray, 1842, belongs to the monotypic genus Soriculus, which is distributed mainly in the Himalayan region. Previous authors have studied its classification based on morphological and molecular data. However, no comprehensive study of the diversity and phylogeny of this species has been performed. In this study, we investigated the molecular phylogeny, genetic diversity, and species divergence of S. nigrescens based on one mitochondrial gene and three nuclear genes. A total of 124 samples from 27 sites in Southwest China were analyzed. Our molecular phylogenetic analyses and species divergence reveal non-monophyly of Soriculus, potentially representing two genera and three clades. Populations from Yunnan (Clade YN) represent the subspecies S. n. minors and should recover the full species status. Populations from Himalayas (Clade A) represent the species S. nigrescens, while populations from southeastern Nyenchen Tanglha Mountains and southern Himalayas (Clade B) represent a new cryptic and unnamed species. Species delimitation analyses and deep genetic distance analysis clearly support the species status of these three evolving clades. The putative new genus and cryptic species should be studied and identified in the future using a more extensive sampling combined with a comprehensive morphological and phylogenetic analysis.

The discrimination of expressions in facial movements by infants: A study with point-light displays.

Perceiving facial expressions is an essential ability for infants. Although previous studies indicated that infants could perceive emotion from expressive facial movements, the developmental change of this ability remains largely unknown. To exclusively examine infants' processing of facial movements, we used point-light displays (PLDs) to present emotionally expressive facial movements. Specifically, we used a habituation and visual paired comparison (VPC) paradigm to investigate whether 3-, 6-, and 9-month-olds could discriminate between happy and fear PLDs after being habituated with a happy PLD (happy-habituation condition) or a fear PLD (fear-habituation condition). The 3-month-olds discriminated between the happy and fear PLDs in both the happy- and fear-habituation conditions. The 6- and 9-month-olds showed discrimination only in the happy-habituation condition but not in the fear-habituation condition. These results indicated a developmental change in processing expressive facial movements. Younger infants tended to process low-level motion signals regardless of the depicted emotions, and older infants tended to process expressions, which emerged in familiar facial expressions (e.g., happy). Additional analyses of individual difference and eye movement patterns supported this conclusion. In Experiment 2, we concluded that the findings of Experiment 1 were not due to a spontaneous preference for fear PLDs. Using inverted PLDs, Experiment 3 further suggested that 3-month-olds have already perceived PLDs as face-like stimuli.

Revalidation and expanded description of Mustela aistoodonnivalis (Mustelidae: Carnivora) based on a multigene phylogeny and morphology.

The lacked-teeth pygmy weasel, Mustela aistoodonnivalis Wu & Kao, 1991, was originally described as being from Taibai Mountain and Zhashui county, Shaanxi, China. Subsequently, it was considered a subspecies or synonym of Mustela nivalis. In a faunal survey of northwestern Sichuan, eight specimens of M. aistoodonnivalis were collected. A molecular phylogenetic analysis of one mitochondrial and six nuclear genes clustered the specimens as a distinct clade and not with M. nivalis. Morphologically, the lack of the second lower molar differentiated them from M. nivalis, and genetic distances were typical of discrete species. These analyses confirmed that M. aistoodonnivalis is an independent species in the genus Mustela.

Assessing human exposure to phthalate esters in drinking water migrated from various pipe materials and water filter elements during water treatments and storage.

Plastic water-supply pipes and filter element are frequently used in municipal water supply systems. Leaching of phthalate esters (PAEs) from these pipes and filter elements to drinking water has become a common concern among the public. In this study, the migrations of 16 phthalate esters (PAEs) in seven different kinds of water-supply product materials were investigated. Di-n-butyl phthalate (DBP) had the highest detection frequency of 54.4% in the water leaching samples of various water supply pipes and water filter elements samples, followed by Diisobutyl phthalate (DIBP, 46/90, 51.1%). The maximum detected concentration level for di(2-ethylhexyl) phthalate (DEHP), diethyl phthalate (DEP), and DBP in the leaching experiment was below the regulatory limit values of 8 µg/L, 300 µg/L, and 3 µg/L for each compound in China standards for drinking water quality. The increasing of the water temperature, the lower pH of the water, and the increasing of the leaching time will increase the migration of PAEs from plastic pipes into water. The chronic daily intake of children aged < 1-12 years to PAEs through drinking water was higher than the rest of the population groups. Carcinogenic risks (CR) of DEHP via drinking water were neglectable for most groups of people, while for young children with age of 1-2 years old, the CR is an acceptable risk.

The Threat of Adversarial Attack on a COVID-19 CT Image-Based Deep Learning System.

The coronavirus disease 2019 (COVID-19) rapidly spread around the world, and resulted in a global pandemic. Applying artificial intelligence to COVID-19 research can produce very exciting results. However, most research has focused on applying AI techniques in the study of COVID-19, but has ignored the security and reliability of AI systems. In this paper, we explore adversarial attacks on a deep learning system based on COVID-19 CT images with the aim of helping to address this problem. Firstly, we built a deep learning system that could identify COVID-19 CT images and non-COVID-19 CT images with an average accuracy of 76.27%. Secondly, we attacked the pretrained model with an adversarial attack algorithm, i.e., FGSM, to cause the COVID-19 deep learning system to misclassify the CT images, and the classification accuracy of non-COVID-19 CT images dropped from 80% to 0%. Finally, in response to this attack, we proposed how a more secure and reliable deep learning model based on COVID-19 medical images could be built. This research is based on a COVID-19 CT image recognition system, which studies the security of a COVID-19 CT image-based deep learning system. We hope to draw more researchers' attention to the security and reliability of medical deep learning systems.

The complete mitochondrial genome of the long-tailed mole: Scaptonyx fusicaudus (Eulipotyphla, Talpidae).

The long-tailed mole (Scaptonyx fusicaudus) belongs to a monotypic genus within the family Talpidae. It is a small semi-fossorial mammal mostly distributed in south-western China. In this study, we obtained the complete mitochondrial genome of S. fusicaudus. The genome is a total of 16,602 bp in length, containing 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNA), 2 ribosomal RNA genes (rRNA) and 2 non-coding regions, with a base composition of 33.51% A, 28.73% T, 23.68% C and 14.08% G. The nucleotide sequence data of 13 protein-coding genes of S. fusicaudus and other 14 insectivora species were used to reconstruct a Bayesian phylogenetic tree. The tree shows that S. fusicaudus belongs to the subfamily Talpinae and is closely related to Urotrichus talpoides.

A Privacy-Preserving Medical Data Sharing Scheme Based on Blockchain.

With the increasing penetration of the Internet of things (IoT) into people's lives, the limitations of traditional medical systems are emerging. First, the typical way of handling sensitive information can easily lead to privacy disclosure. Second, the medical system is relatively isolated. It is difficult for one medical system to share data with another, and the scope of users' activities is limited within the system boundary. To solve these two problems, we propose a new privacy-preserving medical data-sharing scheme by introducing the authorization mechanism and attribute-based encryption (ABE) based on blockchain, which breaks system boundaries and realizes data sharing among several medical institutions. ABE is used to realize scalable access control. In addition, doctors can share their knowledge to diagnose users by introducing many-to-many matching, which means that patients' health data can be represented by multiple keywords and doctors' expertise can be represented by multiple interests. We provide the correctness and security analysis of our scheme and implement a prototype tool on Ethereum. The experimental results show that our scheme solves the contradiction between the privacy preservation of medical data and the necessity of data sharing.

Occurrence and exposure risk assessment of pesticide residues in green tea samples cultivated in Hangzhou area, China.

The concentration of pesticide residues in 105 green tea samples grown in Hangzhou area were investigated. Of the 14 pesticides analysed using gas chromatography-tandem mass spectrometry and the 27 pesticides analysed using liquid chromatography-tandem mass spectrometry, only 18 were detected in the tea samples. The most frequently detected pesticide residues were imidacloprid (35.2%), acetamiprid (26.7%), carbendazim (21.0%), bifenthrin (21.0%), and cyhalothrin (19.1%). Carbofuran was the only pesticide which exceeded in one sample the maximum residue limit. The concentrations of the analytes in tea samples ranged from below the limit of detection (LOD) to 2.64 mg/kg. Their mean concentrations were all below the LOD, except for imidacloprid, acetamiprid, carbendazim, bifenthrin and cyhalothrin. Based on a preliminary long-term exposure assessment, the hazard quotient values of the detected pesticides varied in the range 0.47 × 10-3 - 1.1 × 10-3%, which indicates that these levels did not pose a risk to human health in Hangzhou area.

Addressing the ACGME Competency of Communicating with the Public through the Use of TEDx Talks.

OBJECTIVES: Competency-based public communication skills are not systematically taught in most medical curricula, reflecting a gap between medical knowledge and holistic patient care as trainees transition into clinicians. We sought to investigate the efficacy of technology, entertainment, and design (TED) talks in postgraduate year (PGY) training programs. METHODS: The authors organized an official internal TEDx event in which six PGY trainees volunteered as speakers. Two experienced physicians and two administrators also participated as speakers to provide trainees a didactic shadow learning experience. The remaining PGY trainees, along with clerks, physicians, nurses, pharmacists, and administrators, attended the conference. Before the event, speakers undertook individual training sessions and learned the principles of the presentation structure and storytelling mode. At the end of the event, a survey evaluating overall satisfaction in communication skills and professionalism was administered to all of the attendees. RESULTS: Survey participants totaled 104, with a response rate of 97.2%. TEDx talks improved trainees' levels of patient care, communication, and professionalism. Speakers reported the high level of satisfaction with the event (mean 4.96 on a 5-point Likert scale; standard deviation 0.20). Participants agreed that the shadowing experience was useful and that the event encouraged them to pursue interests outside the medical field. CONCLUSIONS: This study highlights that TED is successful in terms of participant satisfaction and training in communication and professionalism. Engaging and training PGY trainees through TED-style events could bridge the gap between acquired knowledge and professional competencies. The authors recommend the implementation of TED-style events in medical training programs.

Evolution and diversification of Mountain voles (Rodentia: Cricetidae).

The systematics of the Cricetid genus Neodon have long been fraught with uncertainty due to sampling issues and a lack of comprehensive datasets. To gain better insights into the phylogeny and evolution of Neodon, we systematically sampled Neodon across the Hengduan and Himalayan Mountains, which cover most of its range in China. Analyses of skulls, teeth, and bacular structures revealed 15 distinct patterns corresponding to 15 species of Neodon. In addition to morphological analyses, we generated a high-quality reference genome for the mountain vole and generated whole-genome sequencing data for 47 samples. Phylogenomic analyses supported the recognition of six new species, revealing a long-term underestimation of Neodon diversity. We further identified positively selected genes potentially related to high-elevation adaptation. Together, our results illuminate how climate change caused the plateau to become the centre of Neodon origin and diversification and how mountain voles have adapted to the hypoxic high-altitude plateau environment.

Phylogenomics reveals the evolution, biogeography, and diversification history of voles in the Hengduan Mountains.

The Hengduan Mountains (HDM) of China are a biodiversity hotspot whose temperate flora and fauna are among the world's richest. However, the origin and evolution of biodiversity in the HDM remain poorly understood, especially in mammals. Given that the HDM shows the highest richness of vole species in the world, we used whole-exome capture sequencing data from the currently most comprehensive sampling of HDM voles to investigate their evolutionary history and diversification patterns. We reconstructed a robust phylogeny and re-estimated divergence times of the HDM voles. We found that all HDM voles could be divided into a western lineage (Volemys, Proedromys, and Neodon) and an eastern lineage (Caryomys and Eothenomys), and the two lineages originated from two migration events from North Eurasia to the HDM approximately 9 Mya. Both vole lineages underwent a significant acceleration of net diversification from 8-5 Mya, which was temporally congruent with the orogeny of the HDM region. We also identified strong intertribal gene flow among the HDM voles and hypothesized that frequent gene flow might have facilitated the speciation burst of the HDM voles. Our study highlights the importance of both environmental and biotic factors in shaping the biodiversity of mammals in mountain ecosystems.

Molecular phylogeny and taxonomy of the genus Nectogale (Mammalia: Eulipotyphla: Soricidae).

The elegant water shrew, Nectogale elegans, is one of the small mammal species most adapted to a semi-aquatic lifestyle. The taxonomy of the genus Nectogale has received little attention due to difficulties in specimen collection. In this study, we sequenced one mitochondrial and eight nuclear genes to infer the phylogenetic relationship of Nectogale. Phylogenetic analyses revealed two large clades within Nectogale. One clade represented N. elegans, and the other was regarded as N. sikhimensis. The split between N. elegans and N. sikhimensis dated back to the early Pleistocene (2.15 million years ago [Ma]), which might be relevant to the Qinghai-Tibet Plateau (QTP) uplift. The morphological comparison showed several distinguishing characters within Nectogale: the shape of the mastoids, the first lower unicuspid (a1), and the second upper molar (M2). Overall, the molecular and the morphological evidences supported that the genus Nectogale consists of two valid species: N. elegans and N. sikhimensis.

Genomics and morphometrics reveal the adaptive evolution of pikas.

Pikas (Lagomorpha: Ochotonidae) are small mouse-like lagomorphs. To investigate their adaptation to different ecological environments during their dispersal from the Qinghai-Xizang (Tibet) Plateau (QTP), we collected 226 pikas and measured 20 morphological characteristics and recorded habitat information. We also sequenced the genome of 81 specimens, representing 27 putative pika species. The genome-wide tree based on 4 090 coding genes identified five subgenera, i.e., Alienauroa, Conothoa, Lagotona, Ochotona, and Pika, consistent with morphometric data. Morphologically, Alienauroa and Ochotona had similar traits, including smaller size and earlier divergence time compared to other pikas. Consistently, the habitats of Alienauroa and Ochotona differed from those of the remaining subgenera. Phylogenetic signal analysis detected 83 genes significantly related to morphological characteristics, including several visual and hearing-related genes. Analysis of shared amino acid substitutions and positively selected genes (PSGs) in Alienauroa and Ochotona identified two genes, i.e., mitochondrial function-related TSFM (p.Q155E) and low-light visual sensitivity-related PROM1 (p.H419Y). Functional experiments demonstrated that TSFM-155E significantly enhanced mitochondrial function compared to TSFM-155Q in other pikas, and PROM1-419Y decreased the modeling of dynamic intracellular chloride efflux upon calcium uptake. Alienauroa and Ochotona individuals mostly inhabit different environments (e.g., subtropical forests) than other pikas, suggesting that a shift from the larger ancestral type and changes in sensory acuity and energy enhancement may have been required in their new environments. This study increases our understanding of the evolutionary history of pikas.

Multi-locus phylogeny and species delimitations of the striped-back shrew group (Eulipotyphla: Soricidae): Implications for cryptic diversity, taxonomy and multiple speciation patterns.

The striped-back shrew group demonstrates remarkable variation in skull and body size, tail length, and brightness of the dorsal stripe; and karyotypic and DNA variation has been reported in recent years. In this study, we investigated the phylogenetic structure of the group, as well as speciation patterns and demographic history in Mountains of Southwestern China and adjacent mountains, including the southern Himalayas, Mts. Bashan, Wushan, and Qinling. We sequenced a total of 462 specimens from 126 localities in the known range of the group, which were sequenced and analyzed based on 6.2 kb of sequence data from two mitochondrial, six nuclear, and two Y chromosome markers. Phylogenetic analyses of the concatenated mtDNA data revealed 14 sympatric and independently evolving lineages within the striped-back shrew group, including Sorex bedfordiae, S. cylindricauda, S. excelsus, S. sinalis and several cryptic species. All concatenated data (ten genes) showed a consistent genetic structure compared to the mtDNA lineages for the group, whereas the nuclear and the Y chromosome data showed a discordant genetic structure compared to the mtDNA lineages for the striped-back shrew group. Species delimitation analyses and deep genetic distance clearly support the species status of the 14 evolving lineages. The divergence time estimation suggested that the striped-back shrew group began to diversify from the middle Pleistocene (2.34 Ma), then flourished at approximately 2.14 Ma, followed by a series of rapid diversifications through the Pleistocene. Our results also revealed multiple mechanisms of speciation in the Mountains of Southwestern China and Adjacent Mountains with complex landscapes and climate. The uplifting of the Qinghai-Tibetan Plateau, Quaternary climate oscillations, riverine barriers, ecological elevation gradients, topographical diversity, and their own low dispersal capacity may have driven the speciation, genetic structure, and phylogeographic patterns of the striped-back shrew group.