Colour Variation in the Crocodile Lizard (Shinisaurus crocodilurus) and Its Relationship to Individual Quality.

Colour plays a key role in animal social communication including as an indicator of individual quality. Using spectrophotometry, we examined colour variation in the throat and venter of the crocodile lizard (Shinisaurus crocodilurus), an endangered species native to southern China and northern Vietnam. We detected two broad colour variants, individuals with and without red, for each body region and each sex. A cluster analysis of spectral colour measurements (hue, chroma, luminance) revealed discrete throat and ventral morphs when measured in a single snapshot in time. However, photographic evidence revealed that the amount of red relative to body size increased as they got older. Individuals with red were equally likely to be male or female and throat colour was unrelated to ventral colour. Therefore, it is premature to claim that crocodile lizards have discrete colour morphs. We used visual modelling to show that the throat and venter were easily discriminable to a lizard visual system, suggesting they function in social communication. We also asked whether colour variation signalled individual quality. Females with red throats had greater bite force while males with red throats were older. In addition, females with red venters had larger heads. We also detected differences in morphology linked to colour. Females with red throats had slender bodies and longer tails, while individuals lacking red on their throats were stouter and had shorter tails. Finally, throat and ventral colour were unrelated to reproductive output (litter size and mass) in females. Males with greater ventral luminance contrast sired offspring from litters with greater litter mass (including stillborns), while males with greater ventral chromatic contrast sired offspring whose collective live mass (excluding stillborns) was greater. Males with greater luminance contrast also sired more live offspring (excluding stillborns). Collectively, these results suggest that male ventral colour signals individual quality in males. Conservation initiatives should take colour variation into account when planning future captive breeding and release programs for this endangered species.

The Associations of Cerebrospinal Fluid Ferritin with Neurodegeneration and Neuroinflammation Along the Alzheimer's Disease Continuum.

BACKGROUND: Increasing evidence has suggested that iron accumulation plays an important role in the onset and development of Alzheimer's disease (AD). However, the potential mechanism remains unclear. OBJECTIVE: The present study investigated the associations of cerebrospinal fluid (CSF) ferritin, an indicator for brain iron load, with neurodegenerative and inflammatory changes in AD. METHODS: The study involved 302 participants from the Alzheimer's Disease Neuroimaging Initiative (ADNI). They were classified as normal controls (A-T-N-, n = 48), AD continuum (A+TN-, n = 46; A+TN+, n = 166), and suspected non-AD pathology (A-TN+, n = 42), according to the amyloid/tau/neurodegeneration (ATN) system. Group comparisons of CSF ferritin among groups were performed using one-way ANOVA. Linear regression models were used to test the relationships between CSF ferritin and cognitive assessments, and the associations between CSF ferritin and other biomarkers, respectively. RESULTS: We found that CSF ferritin showed significant differences among the ATN groups, with higher concentration in more advanced categories (A+TN+). Furthermore, CSF ferritin level was independently related to cognitive performance (MMSE, ADAS-Cog13, and ADNI-mem). Linear regression analysis indicated positive relationships between CSF ferritin and phosphorylated tau and total tau, rather than Aß42. Significant associations were revealed between CSF ferritin and inflammatory proteins, including TNF-α, TNFR1, TNFR2, ICAM1, VCAM1, TGF-ß1, IL-9, and IP-10, respectively. CONCLUSION: Our results provide new insight into iron dysfunction in AD pathology and highlight elevated brain iron as a possible mechanism of neurodegeneration and neuroinflammation along AD continuum.

Identification of Pseudomonas aeruginosa From the Skin Ulcer Disease of Crocodile Lizards (Shinisaurus crocodilurus) and Probiotics as the Control Measure.

The crocodile lizard (Shinisaurus crocodilurus) is an endangered ancient reptile species. Captive breeding is an important conservation measure for the potential restoration and recovery of their wild populations. However, a skin ulcer disease caused by an unknown pathogen has become a serious threat to captive breeding individuals. In the current study, based on microbial isolation, we identified Pseudomonas aeruginosa as the dominant pathogen in skin ulcer disease. Chinese skinks (Plestiodon chinensis) were used to verify the pathogenicity of P. aeruginosa in skin ulcer disease in vivo. As expected, subcutaneous inoculation of P. aeruginosa induced skin disease in healthy skinks and P. aeruginosa was re-isolated from the induced skin ulcers. Therefore, P. aeruginosa, an opportunistic and ubiquitous pathogen that causes a wide range of infections, appears to be the main pathogen of the skin disease affecting crocodile lizards. In the aquaculture industry, probiotics are widely used in the prevention and control of animal diseases caused by such pathogens. Here, we administered probiotics to the breeding crocodile lizards for 6 months. The three experiment groups treated with different kinds of probiotics showed significance at controlling case incidence. Three of the four groups treated with probiotics showed significant disease prevention (Effective Microorganisms mixed probiotics P = 0.0374; Double-dose Effective Microorganisms, P = 0.0299; Bacillus subtilis, P = 0.0140, T-test), and CFUs in the water of the breeding enclosures were also inhibited after probiotics usage (P < 0.001, T-test). Our study demonstrated the role of Pseudomonas aeruginosa in development of skin ulcer disease of crocodile lizards in a local zoo and offered the probiotic-based method for control measurements, which would be of benefit for the conservation of endangered reptiles.

Skin Microbiota Was Altered in Crocodile Lizards (Shinisaurus crocodilurus) With Skin Ulcer.

Skin diseases commonly affect reptiles, but their relationships to the closely related skin microbiome are not well-understood. In recent years, both the wild and captive populations of the crocodile lizard, a Class I protected endangered animal in China, have suffered serious skin diseases that hamper the rescue and release projects for their conservation. This study conducted a detailed prevalence investigation of a major dermatosis characterized by foot skin ulcer in crocodile lizards. It should be noticed that skin ulcer has been prevalent in both captive and wild populations. There was positive correlation between skin ulcer and temperature, while no significant relationship between skin ulcer and humidity, sex, and age. We further studied the relationship between skin ulcer and the skin microbiota using meta-taxonomics. Results showed that the skin microbiota of crocodile lizards was significantly different from those of the environmental microbial communities, and that skin microbiota had a significant relationship with skin ulcer despite the impact of environment. Both bacterial and fungal communities on the ulcerated skin were significantly changed, which was characterized by lower community diversity and different dominant microbes. Our findings provide an insight into the relationship between skin microbiota and skin disease in reptile, serving as a reference for dermatological etiology in wildlife conservation.

Subtypes of social withdrawal and mental health trajectories during COVID-19 pandemic.

The outbreak of Coronavirus Disease 2019 (COVID-19) has pervasive implications for the well-being of people, especially for the social withdrawn individuals. The present study examined changes of well-being among people in distinct subgroups of social withdrawal - shyness, unsociability, and social avoidance -in different phases of the COVID-19 pandemic using six-wave longitudinal data in China (N = 222; 54.50% female). Results showed that, in general, well-being sharply decreased from the initial phase to the peak phase of the pandemic, but steadily recovered after the peak phase. People in different withdrawal groups displayed different levels and trajectories of well-being during a period of six months. The current study has implications for developing targeted interventions for vulnerable people in public health crisis.

Correction to: Well­Being Reduces COVID­19 Anxiety: A Three­Wave Longitudinal Study in China.

[This corrects the article DOI: 10.1007/s10902-021-00385-2.].

[Analysis of RUNX2 gene variant in a Chinese patient with cleidocranial dysplasia].

OBJECTIVE: To explore the genetic basis for a Chinese patient featuring cleidocranial dysplasia(CCD). METHODS: Genomic DNA was extracted from peripheral blood samples of the patient and his parents. Whole exome sequencing (WES) was carried out for the patient, and suspected variant was verified by Sanger sequencing. RESULTS: WES has identified a missense c.460G>T (p.Val154Phe) (GRCh37/hg19) variant of the RUNX2 gene. The variant was located in the Runt domain, a highly conserved region (PM1); it was not present in either the Genome Aggregation Database or the 1000 Genomes Project (PM2), and was predicted to have a deleterious effect on the gene product by multiple in silico prediction tools (PP3); the clinical phenotype of the patient was highly consistent with that of cleidocranial dysplasia (PP4). Furthermore, the variant was unreported in medical literature and was absent in both parents (PS2). Based on the American College of Medical Genetics and Genomics guidelines, the c.460 G>T variant of RUNX2 gene was predicted to be pathogenic (PS2+PM1+PM2+PP3+PP4). CONCLUSION: The c.460G>T (p.Val154Phe) variant of the RUNX2 gene probably underlay the clinical phenotype in the patient. Above finding has enabled accurate diagnosis and expanded the spectrum of RUNX2 variants.

Well-Being Reduces COVID-19 Anxiety: A Three-Wave Longitudinal Study in China.

The coronavirus disease 2019 (COVID-19) pandemic threatens human beings' livelihoods and mental health, which lowers their well-being and gives rise to anxiety. This study examines whether there is a causal relationship (and, if so, in which direction) between people's well-being and COVID-19 anxiety. Two hundred and twenty-two participants (54.50% female, M age = 31.53, SD = 8.17) from 26 provinces of China completed measures of subjective well-being (SWB) and COVID-19 anxiety at three key nodes of the development of COVID-19 in China. The results showed that people's SWB and COVID-19 anxiety fluctuated with the peak (T1), decline (T2), and trough stages (T3) of the COVID-19 pandemic. Meanwhile, the cross-lagged analysis showed that the participants' SWB at T0 (pre-pandemic stage; the base level of SWB) and T1 could significantly predict their COVID-19 anxiety at T1 and T2 respectively. However, SWB at T2 was not associated with the COVID-19 anxiety at T3. Furthermore, COVID-19 anxiety could not predict subsequent SWB from T1 to T3. The current findings contribute to clarifying the causal relationship between well-being and anxiety through the development of epidemics, as well as finding ways to alleviate people's COVID-19 anxiety. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10902-021-00385-2.

Perceived Social Support Protects Lonely People Against COVID-19 Anxiety: A Three-Wave Longitudinal Study in China.

The isolation necessary to prevent the spread of the coronavirus disease 2019 (COVID-19) can give rise to anxiety, especially for lonely people who often feel upset without others' company. Although isolated from others, people can still receive support from others, which might lower their COVID-19 anxiety. To examine the relationship between loneliness, perceived social support, and anxiety, we measured 222 Chinese participants' (54.50% female, M age = 31.53, SD = 8.17) trait loneliness, chronic anxiety before the outbreak, COVID-19 anxiety at the peak and decline stages of COVID-19, and their perceived social support across the three time points. The results showed that people's perceived social support dramatically increased from the pre-pandemic to the peak COVID-19 stage, and remained stable during the decline of COVID-19 stage. In contrast, COVID-19 anxiety decreased from the peak to the decline stage. Further, perceived social support consistently moderated the relationship between loneliness with both chronic anxiety and COVID-19 anxiety. The current study provides initial evidence that perceived social support provides protection for lonely people in daily life as well as during unexpected disasters, which will contribute to finding ways to alleviate lonely people's anxiety during this global health crisis.

Qualitative Classification of Shear Wave Elastography for Differential Diagnosis Between Benign and Metastatic Axillary Lymph Nodes in Breast Cancer.

Purpose: To examine diagnostic performance of qualitative shear wave elastography (SWE) for evaluation of status of axillary lymph nodes (ALN) in comparison with conventional ultrasonograghy (US) and quantitative SWE parameters. Methods: A total of 118 patients were enrolled, who were all scheduled for breast cancer surgery and core needle biopsy. Conventional US and SWE were performed before biopsy. Based on qualitative evaluation of each ALN, the SWE images were classified into four color patterns: Color Pattern 1: homogeneous; Color Pattern 2: filling defect within lymph node (LN); Color Pattern 3: homogeneous within LN with a localized colored area at the margin; and Color Pattern 4: filling defect within LN with a localized colored area at the margin. The diagnostic performances of the three methods were compared. Results: There were 60 metastatic nodes and 61 benign nodes in the 121 ALNs. Benign ALNs were presented as Color Pattern 1 while metastatic ALNs usually were presented as Color Pattern 2 to 4 (p < 0.05). The AUC of qualitative SWE classification was 0.983, higher than that of quantitative SWE parameters and conventional US (p<0.05). The highest diagnostic performance, with AUC of 0.998, could be achieved if both conventional US and qualitative SWE were applied. Conclusion: The qualitative SWE classification of ALNs proposed in our study exhibited better diagnostic performance than quantitative SWE parameters and conventional US, especially for differentiating metastatic ALNs from benign reactive ALNs. More accurate diagnosis could be reached with this new method and unnecessary biopsy might be avoided in the meantime.

Impacts of maternal characteristics and temperature on juvenile survival in the crocodile lizard: Implications for conservation.

Captive breeding is an important conservation measure that may restore and enhance wild populations of rare and endangered species. Multiple anthropogenic hazards have brought the crocodile lizard, Shinisaurus crocodilurus, to the brink of extinction. We initiated a captive breeding program and quantified female reproductive traits, including reproductive timing, litter size, litter mass, and neonate size. To identify the internal and external factors affecting female reproductive function, we then analyzed how maternal age is related to body size, temperature, and female reproductive traits. We found that larger female crocodile lizards produced more offspring than smaller ones, as both litter size and litter mass were positively related to maternal body size. In contrast, neonate size was independent of maternal body size. Maternal reproductive output varied among different age groups. Young and old females had significantly smaller living litter size and mass than middle-aged females. Among captive females, one-third exhibited early parturition in autumn and winter instead of the following spring, a pattern probably associated with higher ambient temperatures in captivity. Although female reproductive output and neonatal body size did not differ between early- and normal-parturition females, offspring from the former group died earlier than those from the latter. Our study highlights the danger of climate change in hastening parturition, a phenomenon that could significantly hamper neonate survival and impede population recruitment.

Dual-functionalized nanoparticles loaded microbubbles for enhancement of drug uptake.

The application of microbubble (MB)-assisted ultrasound (US) can combine the advantages of real-time imaging and targeted drug delivery. However, the drug loading capacity of MB is limited restricting its application in antitumor procedure. In contrast, nanoparticles (NPs) can carry drugs more efficiently, but adverse side effect induced by unspecific accumulation can not be ignored. Herein, we developed a dual-functionalized NP loaded MB to investigate its potential feasibility for tumor-targeted drug delivery. Firstly, we prepared NPs using heparin as backbone. Targeting ligand folate and cell-penetrating ligand Tat peptide were conjugated to the backbone to deliver paclitaxel (H-F-Tat-P NPs). Subsequently, the dual-functionalized NPs were incorporated with MBs via avidin-biotin linkage to fabricate H-F-Tat-P NPs loaded MBs (NPs-loaded MBs). The combined strategy can take profit of dual functionalities from NPs and sonoporation effect from MBs triggered by US. The prepared NPs have been characterized. The excellent cellular uptake of NPs were qualitative and quantitative analysis by flow cytometry and confocal microscope, the results indicated that it was attributed to not only dual functionalities but also US effect. Foremost, the NPs-loaded MBs combined with US exhibited significant cytotoxicity on both folate receptor (FR) overexpressing and deficiency cells. The combination of dual-functionalized NPs and MBs with US is expected to be a promising strategy for targeted anticancer drug delivery and ultrasound imaging simultaneously.

Diets Alter the Gut Microbiome of Crocodile Lizards.

The crocodile lizard is a critically endangered reptile, and serious diseases have been found in this species in recent years, especially in captive lizards. Whether these diseases are caused by changes in the gut microbiota and the effect of captivity on disease remains to be determined. Here, we examined the relationship between the gut microbiota and diet and disease by comparing the fecal microbiota of wild lizards with those of sick and healthy lizards in captivity. The gut microbiota in wild crocodile lizards was consistently dominated by Proteobacteria (∼56.4%) and Bacteroidetes (∼19.1%). However, the abundance of Firmicutes (∼2.6%) in the intestine of the wild crocodile lizards was distinctly lower than that in other vertebrates. In addition, the wild samples from Guangdong Luokeng Shinisaurus crocodilurus National Nature Reserve also had a high abundance of Deinococcus-Thermus while the wild samples from Guangxi Daguishan Crocodile Lizard National Nature Reserve had a high abundance of Tenericutes. The gut microbial community in loach-fed crocodile lizards was significantly different from the gut microbial community in the earthworm-fed and wild lizards. In addition, significant differences in specific bacteria were detected among groups. Notably, in the gut microbiota, the captive lizards fed earthworms resulted in enrichment of Fusobacterium, and the captive lizards fed loaches had higher abundances of Elizabethkingia, Halomonas, Morganella, and Salmonella, all of which are pathogens or opportunistic pathogens in human or other animals. However, there is no sufficient evidence that the gut microbiota contributes to either disease A or disease B. These results provide a reference for the conservation of endangered crocodile lizards and the first insight into the relationship between disease and the gut microbiota in lizards.

Yin-Cold or Yang-Heat Syndrome Type of Traditional Chinese Medicine Was Associated with the Epidermal Growth Factor Receptor Gene Status in Non-Small Cell Lung Cancer Patients: Confirmation of a TCM Concept.

Traditional Chinese Medicine (TCM) therapies should be tailored according to the different syndrome types. In order to identify the relationship between the TCM Yin-cold (YC) or Yang-heat (YH) syndrome types and the EGFR gene status, we prospectively studied 310 NSCLC patients. TCM YH or YC was diagnosed by three TCM experts. TCM symptoms and signs were entered into a binary cluster analysis. The relationships between the EGFR gene status, YH or YC syndrome types, and classification by cluster analysis were analyzed using the chi-square test and multivariate logistic regression. In the 299 patients who had their EGFR gene tested, 45.24% YC (76/168) and 25.95% YH (34/131) patients had EGFR mutations (p = 0.001). Among the 292 patients entered into the cluster analysis, 132 were classified into group A, with signs and symptoms similar to YC, whereas 160 group B patients were similar to YH. In the 281 patients with EGFR tested, 45.67% group A (58/127) and 28.57% group B patients (44/154) had EGFR mutations (p = 0.003). The EGFR status was independently correlated with TCM syndrome type and classification by cluster analysis on multivariate logistic regression. NSCLC patients with YC were more likely to have EGFR gene mutations.

Body massage performance investigation by brain activity analysis.

Massage has been widely applied to improve health and reduce stress. However, the performance difference between hands-on treatment and treatment by mechanical devices has been little mentioned. Therefore, the main aim of this paper is to investigate a subject's EEG performance under massage treatment applied by hand and treatment applied by mechanical devices. Massage was applied to four acupoints for three minutes each. The massage acupoint sequence was from left Jian-wai-yu, right Jian-wai-yu, left Zuo-fei-yu, and finally right Zuo-fei-yu. An EEG system of 32 channels was used. Twenty-four volunteers, mainly college students, were enrolled. EEG rhythm powers of each massage sessions were derived. Two-way ANOVA revealed that there were also significant interactions between the massage stage and the massage type on delta (P < 0.01), theta (P < 0.05), and beta rhythms (P < 0.01), and there were significant differences at different stages for the mechanical massage group (F = 5.557, P < 0.01). The mechanical massage group had more significant differences than the hands-on group for stage coherence of around coherence on alpha rhythm. Further rhythm power scalp topography between two massage methods is also investigated.

Performance of the five-point grip pen in three screen-based tasks.

This study examined how user handbreadth, brace shape and brace size of a Five-Point Grip Pen affects the performance of three screen tasks: writing, point-and-clicking, and drawing. Nine braces with three shapes (ball, ball-and-brick, and natural) and three brace sizes (large, medium and small) were tested for performance time, number of errors and subjective preference by 32 subjects (16 males and 16 females). The subjects were selected to fit four groups of handbreadth (below 25%ile, 25%ile-50%ile, 50-75%ile and above 75%ile) of Taiwanese laborers. In general, the subjective ranking results were consistent with the results of performance measures in all three tasks. All measures were affected by brace shape and brace size but differently for each task. The study results can be summarized as follows: (1) The natural shape was the best brace for all tasks. (2) For the writing task, the small brace was best for all four handbreadth groups, the medium brace was adequate only for the handbreadth group of above 75%ile, and the large brace was worst for all groups. (3) For the drawing task, the large and medium braces were suitable for all handbreadth groups, while the small brace was not suitable for the handbreadth group of above 75%ile. (4) For point-and-clicking task, the large size was the best brace.

Performance study on touch-pens size in three screen tasks.

Experiments were aimed at investigating the performance of various sizes of touch-pens. The touch-pens were of three different lengths (80, 110 and 140 mm) and four different diameters (5.5, 8, 11 and 15 mm). Three screen tasks were used for determining a reference of ideal dimensions of a pen-based product. Sixteen subjects used 12 touch-pens to perform pointing-and-clicking, writing and drawing tasks. The subjects were asked to rank their preference after each task in the test. The results showed that the length of touch-pens should be designed to extend beyond the handbreadth when gripped. A touch-pen that facilitates natural pointing-and-clicking, writing and drawing tasks each requires a suitable diameter size. However, these tasks are typically performed together, in particular, the pointing-and-clicking and writing tasks. The recommended diameter for all three combined tasks is 8 mm. This finding indicates that selecting a touch-pen with a diameter of 8 mm and a length of 100 mm or longer is suitable for all pen-based devices.

Design and evaluation approach for increasing stability and performance of touch pens in screen handwriting tasks.

Touch pens allow user to write directly on a computer screen. An ergonomically designed touch pen can improve user performance and reduce musculoskeletal injuries. This study investigates the touch pen use in three sections. First, observations of users' handwriting during three screen tasks: writing, pointing-and-clicking and drawing. Second, examinations of the design and development of touch pens based on observed results and related design theory. Third, this study compares the five-point grip pen (FPGP) with the common touch pen. Study conclusions are as follows: (1) The gesture of gripping a common touch pen for on-screen handwriting is unnatural; that is, users tend to use the wrist, elbow or little finger for support. A user's hand and fingers are in a more natural position when using an FPGP. (2) User performance with the FPGP is superior to that with the common touch pen during three screen tasks. (3) The advantages of the FPGP include: increased hand stability, reduced hand fatigue, enhanced performance, and provided adjustability. (4) This study proposed an FPGP whereby a brace was added to a touch pen to increase the stability during screen-based tasks. The idea of adding a brace can be applied to those tasks requiring higher accuracy or continuous freehand without other limb support tasks, such as wall-painting, surgery operation, and other aiming tasks.