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BACKGROUND Both the quality of sleep and depression impact short-term memory (STM) while they influence each other. However, the underlying mechanisms are not yet clear. Herein, we aimed to explore the mediating effect of depression between sleep quality and STM in older adults and further test the gender differences in this relationship. METHODS A cluster sampling survey was carried out among 903 residents that were aged 60+ years in a semi-closed island of Xiamen, China, in 2019. The subjects' sleep quality and depression were measured using the Pittsburgh Sleep Quality Index (PSQI) and the 15-item Geriatric Depression Scale (GDS-15), respectively. The three-item recall test was utilized to measure STM function. Mediation analyses were performed using the structural equation model and bias-corrected bootstrap method. Next, the Wald-test was utilized to determine gender differences in the pathway. RESULTS Overall, 355 men and 508 women, with a mean age of 69.55 and 70.97, respectively, had valid data on the main variables. Depression was found to mediate the effect of sleep quality on STM (a*b = -0.076, p

The pharmacokinetics of levofloxacin mesylate in healthy adult giant panda is unknown. In this study, the pharmacokinetics of levofloxacin after intramuscular administration at a dose of 2 mg/kg and oral administration at a dose of 3 mg/kg in healthy adult giant pandas was determined. Levofloxacin concentrations in plasma were determined using liquid chromatography. In the levofloxacin intramuscular administration group, the absorption and elimination half-lives of the drug were determined to be 0.123 (range: 0.02) hr and 5.402 (range: 0.70) hr, respectively. In the levofloxacin oral administration group, the absorption and elimination half-lives were determined to be 0.325 (range: 0.02) hr and 7.143 (range: 0.63) hr, respectively. Furthermore, the blood-drug concentration of levofloxacin was found to be above 1 μg/ml after 8 hr of intramuscular administration and above 0.5 μg/ml after 12 hr of oral administration. In this study, HPLC conditions and pretreatment methods of plasma samples were optimized and a detection method was established. Our results indicated that in healthy adult giant pandas, levofloxacin was rapidly absorbed and slowly eliminated. This study will therefore provide to be a guide in veterinary research and will be helpful in the rational use of levofloxacin in giant panda.

Background The gut microbiome is essential for the host’s health and serves as an essential reservoir of antibiotic resistance genes (ARGs). We investigated the effects of different factors, including the dietary shifts and age, on the functional characteristics of the giant panda’s gut microbiome (GPs) through shotgun metagenome sequencing. We explored the association between gut bacterial genera and ARGs within the gut based on network analysis. Results Fecal samples (n=60) from captive juvenile, adult, and geriatric GPs were processed, and variations were identified in the gut microbiome according to different ages, the abundance of novel ARGs and the biosynthesis of antibiotics. Among 667 ARGs identified, nine from the top ten ARGs had a higher abundance in juveniles. For 102 ARGs against bacteria, a co-occurrence pattern revealed a positive association for predominant ARGs with Streptococcus. A comparative KEGG pathways analysis revealed an abundant biosynthesis of antibiotics among three different groups of GPs, where it was more significantly observed in the juvenile group. A co-occurrence pattern further revealed a positive association for the top ten ARGs, biosynthesis of antibiotics, and metabolic pathways. Conclusion Gut of GPs serve as a reservoir for novel ARGs and biosynthesis of antibiotics. Dietary changes and age may influence the gut microbiome’s functional characteristics; however, it needs further studies to ascertain the study outcomes.

Breeding and welfare problems confront many conservation breeding programs. Stereotypies—repetitive, unvarying, functionless behaviours —are common abnormal behaviours that often arise in suboptimal conditions. While the role of stereotypies in welfare assessment is well studied, few investigations address the relationship between stereotypic behaviour and reproduction. We examined the correlation between stereotypic behaviour and reproductive performance in 101 giant pandas (Ailuropoda melanoleuca). High stereotyping males copulated more and produced more cubs, suggesting that highly sexually motivated males were prone to stereotypy but also had high reproductive competence. Female stereotypies were negatively associated with all reproductive measures closely tied to behavioural competence: high stereotyping females were less likely to copulate, less likely to mother-rear cubs, and—probably a result of poor maternal care—had lower cub survival. However, females that exhibited stereotypies were more likely to produce a cub, suggesting stereotypies are tied to behavioural but not physiological competence. High stereotyping female pandas also displayed strong and consistent bias toward production of female offspring while paternal relationship to sex allocation was the reverse. These results are consistent with stress-mediated sex allocation theory. Our findings raise concern about differential reproductive success among high and low stereotyping pandas, and possible genetic adaptation to captivity.

Although several mammals impregnate their fur with environmental odors, a phenomenon termed scent anointing or rubbing, the functional relevance of this behavior often is unclear. One theory is that scent anointing could be a form of scent matching with environmental odors to signal competitiveness and home range occupation. In this study we presented giant pandas with a range of odors to determine whether scent matching could provide a functional explanation for scent anointing in this species. We found that only a musk-based perfume elicited significantly more scent-anointing and scent-marking behavior than control. Males were also significantly more likely to scent-anoint and scent-mark than females. A preference for anointing, but not scent marking, when presented with peppermint (an insecticide) also was revealed. Our results suggest that giant pandas differentially scent-anoint with foreign odors to signal home range occupation, and possibly to repel ectoparasites. We also highlight how chemical signaling of resource-holding potential is likely to play an important role in determining competitive interactions between adult male giant pandas.

The giant panda (GP) was the most endangered species in China, and gut microbiota plays a vital role in host health. To determine the differences of the gut microbiota among the male, female and pregnant GPs, a comparative analysis of gut microbiota in GPs was carried out by 16S rRNA and ITS high-throughput sequencing. In 16S rRNA sequencing, 435 OTUs, 17 phyla and 182 genera were totally detected. Firmicutes (53.6%) was the predominant phylum followed by Proteobacteria (37.8%) and Fusobacteria (7.1%). Escherichia/Shigella (35.9%) was the most prevalent genus followed by Streptococcus (25.9%) and Clostridium (11.1%). In ITS sequencing, 920 OTUs, 6 phyla and 322 genera were also detected. Ascomycota (71.3%) was the predominant phylum followed by Basidiomycota (28.4%) and Zygomycota (0.15%). Purpureocillium (4.4%) was the most prevalent genus followed by Cladosporium (2.5%) and Pezicula (2.4%). Comparative analysis indicated that the male GPs harbor a higher abundance of phylum Firmicutes than female GPs with the contribution from genus Streptococcus. Meanwhile, the female GPs harbor a higher abundance of phylum Proteobacteria than male GPs with the contribution from genus Escherichia/ Shigella. In addition, the shift in bacteria from female to pregnant GPs indicated that phylum Firmicutes increased significantly with the contribution from Clostridium in the gut, which may provide an opportunity to study possible associations with low reproduction of the GPs.

The role of intravenous sodium valproate (iVPA) in acute migraine attack has not been completely established. The aim of this updated review was to evaluate the efficacy and safety of iVPA in patients with acute migraine in the emergency department. We searched the PubMed, Web of Science, and Cochrane Library databases for relevant randomized controlled trials (RCTs). The primary outcome was improvement of headache intensity and headache relief. The need for rescue therapy, recurrence of headache, and number of adverse events was also assessed. Seven double-blinded RCTs involving 682 patients were analyzed. Overall, patients receiving iVPA had less improvement of headache intensity (SMD: -0.39, 95% CI: -0.73 to -0.06, P = .02) and lower rate of headache relief (OR: 0.51, 95% CI: 0.33 to 0.77, P = .002) than those receiving other active comparators. In addition, iVPA increased the odds of rescue therapy compared with other active drugs (OR: 3.76; 95% CI: 1.96 to 7.20, P < .0001). Subgroup analysis showed that iVPA was comparable to dexamethasone, with similar improvement of headache intensity, and recurrence of headache. For migraine without aura, we found no significant difference in headache intensity improvement when iVPA was compared with active comparators (SMD: -0.00, 95% CI: -0.54 to 0.54, P = 1.00). iVPA was inferior to the studied comparators and was comparable to dexamethasone for aborting migraine attack. Based on the available evidence, iVPA may be a reasonable alternative or salvage therapy. In particular, iVPA might be a promising agent for migraine with aura and migraine status.

The gut microbiota diversity of eight panda cubs was assessed during a dietary switch. Gut microbiota diversity of panda cubs significantly decreased after bamboo consumption. Carnivorous species living on a plant-based diet possess low microbial diversity. Mice were fed a bamboo diet but did not display low gut microbiota diversity. Giant pandas have an exclusive diet of bamboo; however, their gut microbiotas are more similar to carnivores than herbivores in terms of bacterial composition and their functional potential. This is inconsistent with observations that typical herbivores possess highly diverse gut microbiotas. It is unclear why the gut bacterial diversity of giant pandas is so low. Herein, the dynamic variations in the gut microbiota of eight giant panda cubs were measured using 16S rRNA gene paired-end sequencing during a dietary switch. Similar data from red panda (an herbivorous carnivore) and carnivorous species were compared with that of giant pandas. In addition, mice were fed a high-bamboo diet (80% bamboo and 20% rat feed) to determine whether a bamboo diet could lower the gut bacterial diversity in a non-carnivorous digestive tract. The diversity of giant panda gut microbiotas decreased significantly after switching from milk and complementary food to bamboo diet. Carnivorous species living on a plant-based diet, including giant and red pandas, possess a lower microbial diversity than other carnivore species. Mouse gut microbiota diversity significantly increased after adding high-fibre bamboo to their diet. Findings suggest that a very restricted diet (bamboo) within a carnivorous digestive system might be critical for shaping a low gut bacterial diversity in giant pandas., The gut microbiota diversity of eight panda cubs was assessed during a dietary switch. Gut microbiota diversity of panda cubs significantly decreased after bamboo consumption. Carnivorous species living on a plant-based diet possess low microbial diversity. Mice were fed a bamboo diet but did not display low gut microbiota diversity.

Aim There is no randomized controlled trial to compare the effectiveness and safety of mechanical thrombectomy (MT) to intravenous thrombolysis in patients with posterior circulation occlusion (PCO). Hence, we firstly performed a meta-analysis to investigate the outcomes of MT in PCO and then compared these outcomes to anterior circulation occlusion (ACO) to provide fundamental data to further studies. Methods We searched the PubMed, EMBASE, and Cochrane Library from dates of inception to June 2019 for relevant studies. Outcomes including functional independence at 90 days, successful recanalization, mortality, symptomatic intracranial hemorrhage (sICH), and futile recanalization were extracted. Results Seven studies involving 474 patients with PCO thrombectomy were analyzed. There was a lower rate of functional independence at 90 days and a higher rate of mortality after thrombectomy in PCO versus ACO (odds ratios (OR) 0.72; 95% confidence interval (CI) 0.57-0.90; OR 2.03; 95% CI 1.30-3.18). Recanalization rates were comparable (OR 1.01; 95% CI 0.62-1.65), but a higher futile recanalization rate was found in basilar artery occlusion (BAO) (OR 1.75; 95% CI 1.30-2.37). There was a lower rate of sICH in MT for patients with PCO versus ACO (OR 0.54; 95% CI 0.29-0.99). Conclusions We found that the outcomes of MT for patients with PCO were poorer than with ACO. On the other hand, MT appears to have lower rates of sICH and to increase successful recanalization. Given the high recanalization rate, MT may serve as an adjunct to standard treatment. The key point to improve outcomes is recognizing reliable factors associated with futile recanalization and optimizing the results of MT. But in view of the different characteristics of posterior circulation stroke and anterior circulation stroke, the results are far from robust.

Gene differential expression studies can serve to explore and understand the laws and characteristics of animal life activities, and the difference in gene expression between different animal tissues has been well demonstrated and studied. However, for the world-famous rare and protected species giant panda (Ailuropoda melanoleuca), only the transcriptome of the blood and spleen has been reported separately. Here, in order to explore the transcriptome differences between the different tissues of the giant panda, transcriptome profiles of the heart, liver, spleen, lung, and kidney from five captive giant pandas were constructed with Illumina HiSeq 2500 platform. The comparative analysis of the intertissue gene expression patterns was carried out based on the generated RNA sequencing datasets. Analyses of Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and protein-protein interaction (PPI) network were performed according to the identified differentially expressed genes (DEGs). We generated 194.52 GB clean base data from twenty-five sequencing libraries and identified 18,701 genes, including 3492 novel genes. With corrected p value 2FoldChange| >2, we finally obtained 921, 553, 574, 457, and 638 tissue-specific DEGs in the heart, liver, spleen, lung, and kidney, respectively. In addition, we identified TTN, CAV3, LDB3, TRDN, and ACTN2 in the heart; FGA, AHSG, and SERPINC1 in the liver; CD19, CD79B, and IL21R in the spleen; NKX2-4 and SFTPB in the lung; GC and HRG in the kidney as hub genes in the PPI network. The results of the analyses showed a similar gene expression pattern between the spleen and lung. This study provided for the first time the heart, liver, lung, and kidney’s transcriptome resources of the giant panda, and it provided a valuable resource for further genetic research or other potential research.

This paper, for the first time, reports on a transducer configuration employing weakly coupled resonators demonstrating significant modal overlap. Existing multimode/mode-localized sensors conventionally operate in a regime where modal overlap is negligible; however, theoretically, the sensitivity of such systems significantly increases as the coupling strength between resonators is reduced, moving the system to a regime described by modal overlap. In this work, operation in the modal overlap regime is made possible with consequent benefits in input-referred resolution and stability. Specifically, a coupled resonator microsystem subject to electrostatic stiffness perturbations shows a sensitivity enhancement of 33682 and an input-referred amplitude ratio stability of 28.2ppb, demonstrating a ∼3x improvement in resolution compared to the resonant frequency shift technique.

Compared with the classic frequency-modulated resonant sensors, the recently emerging sensing scheme based on the mode localization phenomenon of the weakly coupled resonators provides the improved sensitivity by more than 2 orders of magnitude using amplitude ratio as the output metric. However, the inherent loci veeringphenomenon of the amplitude ratio over stiffness perturbation curvegenerates strong or weak nonlinearities in different zones, severelylimiting the linear sensing ranges. In this paper, we propose to use the algebraic summation of the amplitude ratios of two vibrational modes for eliminating the nonlinear errors caused by loci veering, extending the linear sensing range to the fullscale. This new output metric was verified as being very effective to suppress the nonlinear errors using two mode-localized stiffness sensors with 2 and 3 degrees of freedom coupled resonators. This is the first theoretically and experimentally demonstrated full-scale linear output metric for mode-localized sensors, which will push forward more applications of the mode-localized sensing scheme.

Reintroductions and translocations have proven to be effective measures for rescuing small, isolated populations of endangered wildlife. However, the reproductive status of released wildlife is hard to obtain. To date, a giant panda named Luxin is the only translocated giant panda that has successfully bred in the wild. Using data collected from the GPS collar attached to her, we analysed her activity and home range during a breeding year and compared these values with those collected during a non-breeding (control) year. Delivery and mating days can be identified by extremely low levels of activity or even by the absence of activity. The activity of a giant panda with an infant was low, but it increased gradually after delivery. The activity rates during both the delayed implantation period and the infant-caring period were significantly lower in the breeding year than those in the control year. In the breeding year, the home ranges during the delayed implantation period and pregnancy were larger than those in the control year, while the home range during the infant-caring period was much smaller than that in the control year. Our results suggest that GPS collars embedded with activity sensors can be used to monitor the breeding status of released female giant pandas. They can provide valuable information for decision making in future release projects, providing only small disturbances to released giant pandas.

The giant panda ( Ailuropoda melanoleuca) is a high-profile threatened species with individuals in captivity worldwide. As a result of advances in captive animal management and veterinary medicine, the ex situ giant panda population is aging, and improved understanding of age-related changes is necessary. Urine and blood samples were collected in April and July 2015 and analyzed for complete blood count, serum biochemistry, and biochemical and microscopic urine analysis for all individuals sampled ( n = 7, 7-16 yr of age) from giant panda housed at the China Research and Conservation Centre for the Giant Panda in Bifengxia, Sichuan Province, China. Hematology and serum biochemistry values were similar to those previously reported for giant panda aged 2-20 yr and to Species360 (formerly International Species Information System) values. Urine was overall dilute (urine specific gravity range: 1.001-1.021), acellular, and acidic (pH range: 6-7). This is the first report of hematologic and serum biochemistry, with associated urinalysis values, in the giant panda aged 7-16 yr.

While the health effects of the colonization of the reproductive tracts of mammals by bacterial communities are widely known, there is a dearth of knowledge specifically in relation to giant panda microbiomes. In order to investigate the vaginal and uterine bacterial diversity of healthy giant pandas, we used high-throughput sequence analysis of portions of the 16S rRNA gene, based on samples taken from the vaginas (GPV group) and uteri (GPU group) of these animals. Results showed that the four most abundant phyla, which contained in excess of 98% of the total sequences, were Proteobacteria (59.2% for GPV and 51.4% for GPU), Firmicutes (34.4% for GPV and 23.3% for GPU), Actinobacteria (5.2% for GPV and 14.0% for GPU) and Bacteroidetes (0.3% for GPV and 10.3% for GPU). At the genus level, Escherichia was most abundant (11.0%) in the GPV, followed by Leuconostoc (8.7%), Pseudomonas (8.0%), Acinetobacter (7.3%), Streptococcus (6.3%) and Lactococcus (6.0%). In relation to the uterine samples, Janthinobacterium had the highest prevalence rate (20.2%), followed by Corynebacterium (13.2%), Streptococcus (19.6%), Psychrobacter (9.3%), Escherichia (7.5%) and Bacteroides (6.2%). Moreover, both Chao1 and abundance-based coverage estimator (ACE) species richness indices, which were operating at the same sequencing depth for each sample, demonstrated that GPV had more species richness than GPU, while Simpson and Shannon indices of diversity indicated that GPV had the higher bacterial diversity. These findings contribute to our understanding of the potential influence abnormal reproductive tract microbial communities have on negative pregnancy outcomes in giant pandas.

The study of wildlife activity patterns is an effective approach to understanding fundamental ecological and evolutionary processes. However, traditional statistical approaches used to conduct quantitative analysis have thus far had limited success in revealing underlying mechanisms driving activity patterns. Here, we combine wavelet analysis, a type of frequency‐based time‐series analysis, with high‐resolution activity data from accelerometers embedded in GPS collars to explore the effects of internal states (e.g., pregnancy) and external factors (e.g., seasonal dynamics of resources and weather) on activity patterns of the endangered giant panda (Ailuropoda melanoleuca). Giant pandas exhibited higher frequency cycles during the winter when resources (e.g., water and forage) were relatively poor, as well as during spring, which includes the giant panda's mating season. During the summer and autumn when resources were abundant, pandas exhibited a regular activity pattern with activity peaks every 24 hr. A pregnant individual showed distinct differences in her activity pattern from other giant pandas for several months following parturition. These results indicate that animals adjust activity cycles to adapt to seasonal variation of the resources and unique physiological periods. Wavelet coherency analysis also verified the synchronization of giant panda activity level with air temperature and solar radiation at the 24‐hr band. Our study also shows that wavelet analysis is an effective tool for analyzing high‐resolution activity pattern data and its relationship to internal and external states, an approach that has the potential to inform wildlife conservation and management across species.

Post-thawed sperm quality parameters vary across different species after cryopreservation. To date, the molecular mechanism of sperm cryoinjury, freeze-tolerance and other influential factors are largely unknown. In this study, significantly dysregulated microRNAs (miRNAs) and mRNAs in boar and giant panda sperm with different cryo-resistance capacity were evaluated. From the result of miRNA profile of fresh and frozen-thawed giant panda sperm, a total of 899 mature, novel miRNAs were identified, and 284 miRNAs were found to be significantly dysregulated (195 up-regulated and 89 down-regulated). Combined analysis of miRNA profiling of giant panda sperm and our previously published data on boar sperm, 46, 21 and 4 differentially expressed (DE) mRNAs in boar sperm were believed to be related to apoptosis, glycolysis and oxidative phosphorylation, respectively. Meanwhile, 87, 17 and 7 DE mRNAs in giant panda were associated with apoptosis, glycolysis and oxidative phosphorylation, respectively. Gene ontology (GO) analysis of the targets of DE miRNAs showed that they were mainly distributed on membrane related pathway in giant panda sperm, while cell components and cell processes were tied to the targets of DE miRNAs in boar sperm. Finally, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of DE mRNAs indicated that most of these DE mRNAs were distributed in membrane signal transduction-related pathways in giant panda sperm, while those in boar sperm were mainly distributed in the cytokine-cytokine receptor interaction pathway and inflammatory related pathways. In conclusion, although the different freezing extenders and programs were used, the DE miRNAs and mRNAs involved in apoptosis, energy metabolism, olfactory transduction pathway, inflammatory response and cytokine-cytokine interactions, could be the possible molecular mechanism of sperm cryoinjury and freeze tolerance.

microRNA (miRNA) is a small endogenous noncoding RNA molecule that plays multiple roles in regulating most biological processes. However, for China’s national treasure giant panda, a world-famous rare and protected species, reports of its miRNA have been found only in blood and breast milk. To explore the miRNA expression differences between different giant panda tissues, here, we generated the miRNA profiles of five tissues (heart, liver, spleen, lung, and kidney) from four giant pandas with Illumina Hiseq 2500 platform, and filtered the differentially expressed miRNAs (DEmiRs) in each tissue, predicted the target genes of miRNA from each tissue based on the DEmiRs. Then, the GO and KEGG enrichment analysis were conducted using the target genes predicted from DEmiRs in each tissue. The RNA-seq generated an average of 0.718 GB base per sample. A total of 1,256 known miRNAs and 12 novel miRNAs were identified, and there were 215, 131, 185, 83, and 126 tissue-specific DEmiRs filtered in the heart, liver, spleen, lung, and kidney, respectively, including miR-1b-5p, miR-122-5p, miR-143, miR-126-5p, and miR-10b-5p, respectively. The predicted target genes, including MYL2, LRP5, MIF, CFD, and PEBP1 in the heart, liver, spleen, lung, and kidney, respectively, were closely associated with tissue-specific biological functions. The enrichment analysis results of target genes showed tissue-specific characteristics, such as the significantly enriched GO terms extracellular matrix in the heart and insulin-like growth factor binding in the liver. The miRNA profiles of the heart, liver, spleen, lung, and kidney of giant panda have been reported in this study, it reveals the miRNA expression differences between different tissues of the giant panda, and provides valuable genetic resources for the further related molecular genetic research of the rare and protected species giant panda and other mammals.

In order to study the typical vaginal bacterial flora of giant pandas (Ailuropoda melanoleuca), we took vaginal swabs for the sake of bacterial isolation, from 24 healthy female giant pandas. A total of 203 isolates were identified, representing a total of 17 bacterial species. The most common bacteria isolated were Lactobacillus spp. (54.2%, 13 of 24), followed by Staphylococcus epidermidis (41.7%, 10 of 24) and Escherichia coli (33.3%, 8 of 24). Some opportunistic pathogenic bacteria, such as Peptostreptococcus spp., Klebsiella pneumoniae, and Proteus mirabilis, were also isolated but showed no pathology. Antimicrobial susceptibility testing of aerobic bacterial isolates was performed with disk diffusion method. Of the 152 isolates, resistance was most frequently observed with chloramphenicol (17.8%), followed by tetracycline (14.5%), ciprofloxacin (12.5%), streptomycin (11.8%), and florfenicol (11.8%), while 7.2% were multidrug resistant. This is the first report of the normal vaginal culturable bacterial flora of giant pandas, followed by the antimicrobial susceptibility patterns of the isolates.

Background Ovarian cancer is diagnosed clinically by detecting ovarian cancer-related factors and markers. Here, we report a case of giant panda ovarian tumor metastasis with a combination of clinical and histopathological diagnosis. Case presentation Histopathological studies revealed severe lesions and tumor cells in the ovaries, lungs, spleen, kidneys and perianal tissue. Immunohistochemistry staining showed that the ovarian cancer markers B7-H4, CA125, and HE4 were highly expressed in the lungs, kidneys, spleen, ovaries and perianal tissue. Tumor marker tests detected significantly high levels of AFP in serum. Conclusion Clinical biomarkers combined with histopathology can provide a more accurate diagnosis of ovarian cancer metastasis and identification of ovarian cancer types than either method alone. The giant panda’s death may be due to granulosa cell tumor and theca cell tumor metastasis causing multiple organ dysfunction or even failure.

Sperm cryopreservation and artificial insemination are important methods for giant panda breeding and preservation of extant genetic diversity. Lower conception rates limit the use of artificial insemination with frozen-thawed giant panda sperm, due to the lack of understanding of the cryodamaging or cryoinjuring mechanisms in cryopreservation. Long non-coding RNAs (lncRNAs) are involved in regulating spermatogenesis. However, their roles during cryopreservation remain largely unexplored. Therefore, this study aimed to identify differentially expressed lncRNAs and mRNAs associated with cryodamage or freeze tolerance in frozen-thawed sperm through high throughput sequencing. A total of 61.05 Gb clean reads and 22,774 lncRNA transcripts were obtained. From the sequencing results, 1477 significantly up-regulated and 1,396 significantly down-regulated lncRNA transcripts from fresh and frozen-thawed sperm of giant panda were identified. GO and KEGG showed that the significantly dysregulated lncRNAs and mRNAs were mainly involved in regulating responses to cold stress and apoptosis, such as the integral component of membrane, calcium transport, and various signaling pathways including PI3K-Akt, p53 and cAMP. Our work is the first systematic profiling of lncRNA and mRNA in fresh and frozen-thawed giant panda sperm, and provides valuableinsights into the potential mechanism of cryodamage in sperm.

β-Defensins are small antimicrobial proteins expressed in various organisms and have great potential for improving animal health and selective breeding programs. Giant pandas have a distinctive lineage in Carnivora, and it is unclear whether β-defensin genes have experienced different selective pressures during giant panda evolution. We therefore characterized the giant panda (Ailuropoda melanoleuca) β-defensin gene family through gap filling, TBLASTN, and HMM searches. Among 36 β-defensins identified, gastrointestinal disease may induce the expression of the DEFB1 and DEFB139 genes in the digestive system. Moreover, for DEFB139, a significant positive selection different from that of its homologs was revealed through branch model comparisons. A Pro-to-Arg mutation in the giant panda DEFB139 mature peptide may have enhanced the peptide’s antimicrobial potency by increasing its stability, isoelectric point, surface charge and surface hydrophobicity, and by stabilizing its second β-sheet. Broth microdilution tests showed that the increase in net charge caused by the Pro-to-Arg mutation has enhanced the peptide’s potency against Staphylococcus aureus, although the increase was minor. We expect that additional gene function and expression studies of the giant panda DEFB139 gene could improve the existing conservation strategies for the giant panda.

Documenting the different social and behavioural contexts that vocalisations are produced in remains an important step towards understanding the functional relevance of specific call types in a given species' vocal repertoire. In this study we investigated whether seasonal differences and the presence or absence of male and female conspecifics influence the production of male giant panda vocal signals. To this end, captive male giant pandas were observed during and outside of the breeding season in three social contexts: only male conspecific neighbours, only female conspecific neighbours, and a context with no neighbours. We found that males were more likely to bleat, chirp, honk and moan during the breeding season, and showed a tendency to growl more outside of the reproductive period. The contextual analysis revealed that bleats were more likely to be produced by males when opposite-sexed conspecifics are in close attendance during the breeding season. Conversely, males were more likely to chirp when neighboured by males than females or no neighbours. In addition, males were more likely to honk in the absence of neighbouring conspecifics during the breeding season, raising the possibility that these calls function to signal location and gain the attention of potential mates. Moans were produced more often when male giant pandas had male than female neighbours during the breeding season, which may reflect mild aggression towards these same-sexed rivals, whereas the production of barks and growls did not vary according to season or the sex of conspecific neighbours. Our findings underscore the importance of male giant panda bleats for coordinating reproduction and promoting contact with potential mating partners in this non-gregarious species, and yield fresh insights into the function of male honks that warrant further investigation. They also provide a basis for comparison with free-ranging giant panda vocal behaviour that could potentially inform conservation efforts.

Reproductive monitoring for captive breeding in giant pandas is based on behavioural observation and non-invasive hormone analysis. In urine, interpretation of results requires normalisation due to an animal's changing hydration. Correction of urinary concentrations based on creatinine is the gold standard. In this study, a largely unexplored, easy-to-perform normalisation technique, based on urinary specific gravity (USpG), was examined and compared to creatinine. To this extent, six cycles from two female pandas (SB741(1) and SB569 (5)) were monitored through urine analysis for oestrogen, progesterone, ceruloplasmin and 13,14-dihydro-15-keto-PGF2a (PGFM). The Pearson's correlation between creatinine and USpG was high (r = 0.805-0.894; p < 0.01), indicative for a similar performance of both normalisation methods. However, generally lower values were observed during pro-oestrus and primary (progesterone) rise. This could be associated with huge shifts in appetite, monitored by faecal output (kg) with an averaged > 50% decrease during oestrus and > 50% increase during primary progesterone rise. In parallel, respectively highest and lowest creatinine and USpG levels, were measured, with creatinine obviously more affected as a result of linkage with muscle tissue metabolism affected by reproductive hormones. As a consequence, metabolite levels were significantly different between both corrected datasets with significantly higher oestrogen peak levels during oestrus ranging from 2.13-86.93 and 31.61-306.45 ng/mL (USpG correction) versus 2.33-31.20 and 36.36-249.05 ng/mL Cr (creatinine correction) for SB569 and SB741 respectively, and significant lower progesterone levels during primary progesterone rise ranging from 0.35-3.21 and 0.85-6.80 ng/mL (USpG correction) versus 0.52-10.31 and 2.10-272.74 ng/mL Cr (creatinine correction) for SB569 and SB741 respectively. Consequently, USpG correction rendered unbiased profiles, less subject to variation and metabolic artefacts and therefore allowed a more straightforward identification of peak oestrogen and onset of secondary progesterone rise, being potentially advantageous for future studies unravelling key giant panda reproductive events, including (delayed) implantation. The alternative application of USpG as a normalisation factor was further supported by its easy application and environmental and technical robustness.

The giant pandas' (Ailuropoda melanoleuca) reproductive strategy is unique among mammals. Yet there are characteristics of giant panda behaviour we do not fully understand. Probably one of the least understood is the assumption that in captivity virtually all giant panda females rear only one cub when twins are born and abandon the other if given the chance. So far, only two females have raised twins simultaneously, but just with intensive human assistance. This case-study marks the first successful rearing of giant panda twins in captivity entirely by the mother. Using video data for detailed behavioural observations, we provide the first behavioural assessment of a giant panda female raising two cubs simultaneously without direct human assistance or disturbance. We compared the maternal behaviour during the denning period of twin cubs raised in 2016 with two singleton cubs born 2007 and 2010. YANG YANG, the dam, rested less and interacted more with the twins than with the singletons in the first month postpartum and invested a greater part of her daily time budget on rearing the twins. We discuss potential favourable factors for the autonomous twin-rearing of a female giant panda, which could serve as a model for similar efforts elsewhere.

The giant panda Ailuropoda melanoleuca is an endangered species and is a symbol for wildlife conservation. Although efforts have been made to protect this rare and endangered species through breeding and conservative biology, the long-term preservation of giant panda genome resources (gametes, tissues, organs, genomic libraries, etc.) is still a practical option. In this study, the giant panda skeletal muscle-derived cell line was successfully established via primary explants culture and cryopreservation techniques. The population doubling time of giant panda skeletal cells was approximately 33.8 h, and this population maintained a high cell viability before and after cryopreservation (95.6% and 90.7%, respectively). The two skeletal muscle-specific genes SMYD1 and MYF6 were expressed and detected by RT-PCR in the giant panda skeletal muscle-derived cell line. Karyotyping analysis revealed that the frequencies of giant panda skeletal muscle cells showing a chromosome number of 2n=42 ranged from 90.6∼94.2%. Thus, the giant panda skeletal muscle-derived cell line provides a vital resource and material platform for further studies and is likely to be useful for the protection of this rare and endangered species.

Studies on animal space use can reveal insights into how animals interact with one another and their environment. Research on the space use patterns of the endangered giant panda (Ailuropoda melanoleuca) in China has nevertheless lagged behind that of many other species, as a government moratorium prevented telemetry data collection on pandas from 1995 to 2006. We studied 5 giant pandas using GPS telemetry and estimated home ranges, core areas, and space use using model-based approaches. Home range 95% area was 6 km2 for the single male studied and averaged 4.4 ± 1.2 (± SD) km2 for the 4 females. Pandas occupied several small core areas that they revisited after time lags of up to several months. Pandas also displayed significant space use interactions, especially among the male and 2 different females across several weeks during a fall season, a time of year not previously thought to involve extensive inter-panda interaction.

To investigate the associations of serum levels of intercellular adhesion molecule-1 (ICAM1) and the pro-inflammatory cytokine interleukin-6 (IL-6) with migraine and migraine subtypes, and to study their correlation with each other in this condition. We used enzyme-linked immunosorbent assay to measure serum levels of ICAM1 and IL-6 in 103 migraine patients with and without aura, in both attack and pain-free periods, and in 100 healthy control subjects. Serum levels of ICAM1 and IL-6 were significantly higher in migraine patients during attacks than in controls (p < 0.05). Serum ICAM1 levels were significantly higher in migraine with aura (MA) than in migraine without aura (MO), (p < 0.05). Correlation analysis indicated a significant positive correlation between serum levels of ICAM1 and IL-6 (p < 0.05) in migraine patients during attacks. Our results indicate that ICAM1 and IL-6 are involved in the pathogenesis of migraine attacks, possibly via an interactive mechanism.

Fifteen giant pandas (Ailuropoda melanoleuca) from the Chinese Conservation and Research Center for the Giant Panda (CCRCGP) in Bifengxia, Sichuan, China were analyzed for blood lead concentrations (Pb-B) during the 2017 breeding season. Thirteen of the 15 bears showed Pb-B below the method detection limit (MDL) of 3.3 µg/dL. The two remaining bears, although above the MDL, contained very low concentrations of lead of 3.9 and 4.5 µg/dL. All 15 giant pandas in this analysis had Pb-B concentrations that were within normal background concentrations for mammals in uncontaminated environments. For a threatened species, whose native country is plagued by reports of extremely high air pollution, our findings suggest that giant pandas at the CCRCGP are not absorbing lead at concentrations that would adversely affect their health.

The physiological role of miRNAs is widely understood to include fine-tuning the post-transcriptional regulation of a wide array of biological processes. Extensive studies have indicated that exosomal miRNAs in the bodily fluids of various organisms can be transferred between living cells for the delivery of gene silencing signals. Here, we illustrated the expression characteristics of exosomal miRNAs in giant panda breast milk during distinct lactation periods and highlighted the enrichment of immune- and development-related endogenous miRNAs in colostral and mature giant panda milk. These miRNAs are stable, even under certain harsh conditions, via the protection of extracellular vesicles. These findings indicate that breast milk may facilitate the dietary intake of maternal miRNAs by infants for the regulation of postnatal development. We also detected exogenous plant miRNAs from the primary food source of the giant panda (bamboo) in the exosomes of giant panda breast milk that were associated with regulatory roles in basic metabolism and neuron development. This result suggested that dietary plant miRNAs are absorbed by host cells and subsequently secreted into bodily fluids as potential cross-kingdom regulators. In conclusion, exosomal miRNAs in giant panda breast milk may be crucial maternal regulators for the development of intrinsic ‘slink’ newborn cubs.

BACKGROUND The aim of this study was to determine the plasma levels of cyclooxygenase-2 (COX-2) and visfatin in different stages and different subtypes of migraine headaches compared to a control group to elucidate the pathological mechanisms involved. MATERIAL AND METHODS We recruited a case-control cohort of 182 adult migraine patients and 80 age-matched and gender-matched healthy controls. The migraine patients were divided into two groups: the headache-attack-period group (Group A, n=77) and the headache-free-period group (Group B, n=105). The two groups were further divided into subgroups according to whether they had aura symptoms. Solid phase double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was used to measure the plasma levels of COX-2 and visfatin. Statistical analysis was performed using SPSS 17.0. RESULTS The plasma levels of COX-2 and visfatin in the headache-attack-period group were significantly higher than in the headache-free-period group and the control group; there were no significant differences between the headache-free group and the control group. There were no significant differences in plasma levels of COX-2 and visfatin between the subgroups: headache-attack-period with aura subgroup and the headache-attack-period without aura sub group. CONCLUSIONS COX-2 and visfatin participated in the pathogenesis of migraine headaches. The presence of aura had no effect on the serum levels of COX-2 and visfatin.

This paper experimentally demonstrates the ambient pressure drift rejection capability in the full measurement range of the mode-localized sensors. Based on a mode-localized resonant stiffness sensor, the experimental results show that the maximum measurement error of the amplitude ratio readout is ∼2.74% whereas that of the frequency readout is ∼21.63% with a pressure range of [2.6, 20] Pa. And the amplitude ratio based sensitivity is averagely ∼1970.3 times higher than the frequency based sensitivity. It can be concluded that the mode-localized sensors can not only amplify the sensitivity but also reject the ambient pressure drift.

To explore the effects of the mutations of ATP6 and ATP8 genes on energy metabolism and genetic structure, we sequenced the ATP6 and ATP8 genes of Ailuropoda melanoleuca. Our results showed that ATP8 is a conserved gene and ATP6 gene is positively selected during the evolution of the giant panda population with a low genetic diversity. Population expansion was observed in the giant panda group. The T179C mutation on Haplotype7 made the production of a potential phosphorylation site. This non-synonymous mutation may occur at the post-translational modification site that have a potential effect on the function of ATP synthase, related to the maintenance of body temperature of pandas at low metabolic rates.

Wild populations of giant pandas (Ailuropoda melanoleuca) have steadily increased in the past 2 decades, but the species' distribution remains highly fragmented. Since 2009, an introduction program has worked to rescue the giant panda population of Liziping National Nature Reserve in southwestern Sichuan Province, China. Using Global Positioning System and activity collar data collected between May 2011 and March 2016, we investigated the post-release behavior of the first 5 pandas introduced to Liziping, 4 of which were bred in captivity. Using a change-point analysis, we tested several models of post-release adjustment to the habitat. We found that it took 3–4 months for captive-bred individuals to exhibit movement patterns characteristic of their long-term behavior. Furthermore, we found that, for these individuals, post-adjustment behavior varied by season, with activity levels peaking between May and July, a period of high resource availability. This also corresponded with a decrease in large movement events, where individuals were less likely to travel long distances quickly during these months. Unlike wild giant pandas in more northerly reserves, the 5 pandas released in Liziping (both captive-bred and translocated) did not exhibit any seasonal migration between elevations. Finally, we found that our study individuals had 2 daily periods of activity, which was comparable to those reported in the literature for wild individuals. Our results suggest that captive-bred giant pandas are able to successfully adjust to the wild and, after a period of adjustment, settle into long-term behavior patterns.

Translocation is believed to be one of the important methods to protect small and isolated populations of giant pandas (Ailuropoda melanoleuca) from local extinction. Dynamic monitoring of the giant pandas' health statuses after release allows timely rescue measures or termination of the release procedure when necessary, thereby reducing the risk of death. However, it is difficult to diagnose the health of a giant panda after translocation by routine veterinary examinations. Based on data collected from GPS tracking collars, we analyzed the behaviours of six giant pandas after release. We compared the behavioural differences between the giant panda Xuexue (XU), which died due to disease, and four healthy giant pandas after release into the wild to identify a new method to assess the health statuses of giant pandas after release. Our results showed that daily activity levels of the healthy giant pandas were higher than XU after released into the wild; the healthy giant pandas were diurnal, but XU tended to be nocturnal. All healthy giant pandas and XU were not active in the crepuscular phase. These results suggest that daily activity levels and circadian rhythms can be used to assess the health status of giant pandas during the translocation project.

The behavioural adaptation (movement and habitat utilization) of translocated organisms to a new environment after their release is crucial in translocation programs because it may affect survival. Therefore, identifying the factors determining habitat selection by the relocated animals is essential to improving the planning and the outcome of such programs. Using the data from three relocated giant pandas (Ailuropoda melanoleuca), we studied the habitat utilization, release-site fidelity, and interaction with resident giant pandas. The results showed that the quality of habitat used by the relocated giant pandas was significantly higher than the average habitat quality of the research region, and was significantly lower than that used by resident giant pandas. This suggested that the released three giant pandas had habitat selection abilities. The three released giant pandas gradually moved away from their release sites and did not exhibit site fidelity. In the first six months, the giant pandas stayed within about 3 km of their release sites, where habitat was good quality but overlapped with the distribution of resident giant pandas. The overlap of location between released and resident giant pandas decreased after six months when the released giant pandas moved away from their release sites.

We investigated the possible association between two single nucleotide polymorphisms of IL10 (-1082A/G and -592C/A) and susceptibility to ischemic stroke. In total, 335 patients with proven ischemic stroke and 335 control subjects were recruited from Xinxiang Central Hospital between March 2013 and May 2015. The IL10 -1082A/G and -529C/A polymorphisms were investigated by polymerase chain reaction-restriction fragment length polymorphism. When compared with the control subjects, patients with ischemic stroke were more likely to be male, have a habit of tobacco smoking, have higher BMI, have hypertension or diabetes mellitus, and have higher levels of TC, LDL-C, HDL-C, and TG. The multivariate logistic regression analyses revealed that the AA genotype of IL10 -1082A/G was significantly associated with development of ischemic stroke in a Chinese population compared with the GG genotype (OR = 1.93, 95%CI = 1.15-3.25). In the dominant model, the association between GA+AA genotype of IL10 -1082A/G and risk of ischemic stroke was also significant compared with the GG genotype, and the adjusted OR (95%CI) for the GA+AA genotype was 1.41 (1.02-1.94). Thus, our study suggests that IL10 gene polymorphisms contribute to the development of ischemic stroke.

This paper reports a high-sensitive MEMS resonant electrometer based on mode localization of two weakly coupled resonators (WCRs). In the electrometer, the small electric charge input will cause a stiffness perturbation to one resonator of the WCRs, leading to a drastic change of the mode shape owing to the mode localization phenomena. By measuring the shift of the amplitude ratio, the small charge fluctuation can be accurately sensed. The experimental results show that with a charge input of 144fC the relative sensitivity based on the amplitude ratio is 663751ppm, which is −2341 times higher than that of the frequency (283.56ppm).

Artificial insemination (AI) is an important component of captive breeding programs for endangered species, such as the giant panda. The panda has been the subject of increasingly successful captive breeding programs involving a compilation of assisted breeding techniques, including AI using cryopreserved spermatozoa. AI implementation is currently hampered by a lack of understanding of the factors that may cause failure. We investigated factors influencing the probability of success of AI for 14 giant panda females housed at the China Center for Research and Conservation of the Giant Panda (CCRCGP) inseminated in a total of 20 instances using cryopreserved spermatozoa from 11 males currently residing in 6 different captive breeding institutions. One of the pandas was the oldest giant panda female to ever successfully conceive from AI (20.5 years old). The success of AI was significantly affected by the timing of AI in relationship to both timing of peak urinary estrogen of the female and percent decline in urinary estrogen between the peak level and the first AI attempt. Our results suggest that the window for successful AI in giant pandas may be narrower than previously suspected, although individual differences in rates of decline in urinary estrogen may reflect some degree of variation in this crucial window across females. Our results are consistent with recent research on pandas and other species that demonstrates the efficacy of cryopreserved spermatozoa for AI and highlights the need for more in-depth analysis of factors related to female physiology that may influence its success.

Surprisingly little is known about how mammal vocal signals are used to achieve behavioural synchrony in the lead up to copulation. The ability to signal short-term fluctuations in arousal levels and behavioural intention is likely to be particularly important for synchronizing mating behaviour in asocial species, which must overcome their natural avoidance and aggressive tendencies to mate. Here, we examined vocal behaviour during breeding encounters in captive giant pandas ( Ailuropoda melanoleuca ) to gain a greater understanding of how close-range vocal signalling mediates reproduction in this asocial, and conservation-dependent species. Our results revealed that the occurrence of different giant panda vocalizations and acoustic variation within these calls is predictive of successful encounters leading to copulation, as opposed to unsuccessful encounters that do not. In addition, key differences were detected between vocalizations produced during and just prior to copulation. These findings illustrate that vocal exchanges are crucial for achieving behavioural synchrony and signalling intention to mate in giant pandas, and could also provide a valuable tool for breeding programmes, helping conservation managers to assess the likelihood of breeding introductions leading to copulation or potentially injurious failure.

Understory vegetation is an important component in forest ecosystems not only because of its contributions to forest structure, function and species composition, but also due to its essential role in supporting wildlife species and ecosystem services. Therefore, understanding the spatio-temporal dynamics of understory vegetation is essential for management and conservation. Nevertheless, detailed information on the distribution of understory vegetation across large spatial extents is usually unavailable, due to the interference of overstory canopy on the remote detection of understory vegetation. While many efforts have been made to overcome this challenge, mapping understory vegetation across large spatial extents is still limited due to a lack of generality of the developed methods and limited availability of required remotely sensed data. In this study, we used understory bamboo in Wolong Nature Reserve, China as a case study to develop and test an effective and practical remote sensing approach for mapping understory vegetation. Using phenology metrics generated from a time series of Moderate Resolution Imaging Spectroradiometer data, we characterized the phenological features of forests with understory bamboo. Using maximum entropy modeling together with these phenology metrics, we successfully mapped the spatial distribution of understory bamboo (kappa: 0.59; AUC: 0.85). In addition, by incorporating elevation information we further mapped the distribution of two individual bamboo species, Bashania faberi and Fargesia robusta (kappa: 0.68 and 0.70; AUC: 0.91 and 0.92, respectively). Due to its generality, flexibility and extensibility, this approach constitutes an improvement to the remote detection of understory vegetation, making it suitable for mapping different understory species in different geographic settings. Both biodiversity conservation and wildlife habitat management may benefit from the detailed information on understory vegetation across large areas through the applications of this approach.