Population Viability Analysis on Chinese Goral Indicates an Extinction Risk for a Local Population in Beijing, China.

The Chinese goral (Naemorhedus griseus) is identified as a vulnerable species on the Red List of China's Biodiversity and listed as a national second-class key protected wild animal in China. It is a representative flagship ungulate in Beijing. Its distribution range is fragmented and small populations are separated by dense infrastructures and tourism sites. Understanding its population status provides a foundation to plan effective conservation strategies. In this study, a population viability analysis was conducted with VORTEX (10.5.6.0) on a Chinese goral population in Beijing Yunmengshan Nature Reserve with the data collected by camera trapping and parameters referenced from other goral populations. The results show that this population will initially increase in the next 20 years and then decrease with a 32% probability of extinction risk. Supplementation with four adults, two females and two males, every 10 years would help minimize the extinction risk of this population. The results highlight the vital limiting factors for Chinese goral, including the initial population size, sex ratio at birth and mortality of infants (especially female infants). To improve the protection efficiency, detailed population parameters should be further acquired through continuous monitoring of this population. A thorough large-scale study should be carried out on other segregated goral populations in Beijing to facilitate the recovery of this endangered species.

Clinical characteristics and prognosis of early-onset cholangiocarcinoma: a population-based study.

BACKGROUND: Little is known about the disease of early-onset cholangiocarcinoma (EOC). The primary objective of this study was to compare EOC with later-onset cholangiocarcinoma (LOC) concerning clinical features and survival prognosis. METHODS: 19325 cholangiocarcinoma patients were extracted from 1975 to 2020 in the SEER database. Cox regression analysis and Kaplan-Meier survival curves were used for the evaluation of cause-specific survival (CSS) and overall survival (OS). To reduce confounding, we compared survival differences between the EOC and LOC groups using propensity score matching (PSM). RESULTS: 4037 cholangiocarcinoma patients were included in the study, of which 274 were EOC and 3763 were LOC. Early-onset patients were more likely to be non-white, and intrahepatic cholangiocarcinoma. At diagnosis, patients had advanced AJCC stage, lymph node metastase and distant metastase. The EOC patients were more likely to receive surgery, radiotherapy, and chemotherapy than later-onset patients. Multifactorial COX analysis indicated that EOC patients had lower mortality risk than later-onset patients, and similar results were obtained after PSM; Kaplan-Meier survival curves corroborated that early-onset patients exhibited better OS than later-onset patients, and this survival advantage persisted after PSM. Further subgroup analysis following matching demonstrated that early-onset patients had better OS than later-onset patients in the surgical subgroup, while there were no statistically significant differences in the radiotherapy and chemotherapy subgroups. CONCLUSION: The EOC patients typically exhibit an intrahepatic presentation and generally experience a more favorable prognosis. Surgery emerged as a critical treatment modality significantly influencing the overall prognosis of EOC.

Seasonal Variation in Gut Microbiota of the Wild Daurian Ground Squirrel (Spermophilus dauricus): Metagenomic Insights into Seasonal Breeding.

The Spermophilus dauricus, the wild Daurian ground squirrel, is known to exhibit seasonal breeding behavior. Although the importance of gut microbiota in animal digestion, metabolism, and immunity is well-established, the correlation between gut microbiota and seasonal breeding in this species remains inadequately explored. In the present study, using metagenomic sequencing technology, the compositions and functions of the gut microbiota of wild Daurian ground squirrels in different breeding seasons were explored. The dominant gut microbial phyla were Firmicutes and Bacteroidetes. The Firmicutes were predominant in the breeding season, whereas Bacteroidetes were predominant in the non-breeding season. At the genus level, Lactobacillus accumulated during the breeding season, whereas Odoribacter and Alistipes increased during the non-breeding season. GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genome) annotations indicated that genes in gut samples were highly associated with metabolic functions. The differential expression gene analysis showed that genes related to the phosphotransferase system, cysteine, and methionine metabolism were highly expressed during the breeding season, whereas the non-breeding season upregulated genes were enriched in starch and sucrose metabolism and bacterial chemotaxis pathways. In conclusion, this study could provide a reference for investigating gut microbiota in seasonal breeding animals and offer new insight into gut microbial function.

Current Genetic Structure Analysis of Leopard Cats Reveals a Weak Disparity Trend in Subpopulations in Beijing, China.

In the face of habitat shrinkage and segregation, the survival of wild cats looks bleak. Interpreting their population genetic structure during habitat fragmentation is critical in planning effective management strategies. To reveal the segregation effects of road construction and human settlements on the population genetic structure, we analyzed non-invasive fecal DNA samples from leopard cats (Prionailurus bengalensis) from five nature reserves in mountainous areas around Beijing. We focused on microsatellite markers. A total of 112 individual leopard cats were identified among 601 samples of scat, and moderate population genetic diversity was detected. Microsatellite-marker-based genetic differentiation (Fst) and gene flow (Nm) showed a weak trend toward discrepancies in the Baihuashan and Songshan subpopulations, which indicated habitat fragmentation effects on individual dispersal. Because the segregated subpopulations may suffer a high risk of genetic diversity loss, we suggest that their genetic structure be monitored with more molecular markers to detect any changes, and that female individuals be artificially introduced as needed to maintain the viability of the leopard cat subpopulations in Beijing.

Seasonal expressions of GPR41 and GPR43 in the colon of the wild ground squirrels (Spermophilus dauricus).

G-protein-coupled receptor 41 (GPR41) and G-protein-coupled receptor 43 (GPR43) are important short-chain fatty acids (SCFAs) receptors. Previous studies indicated that GPR41 and GPR43 are involved in the secretion of gastrointestinal peptides, and glucose and lipid metabolism, and are closely related to obesity and type II diabetes, and other diseases. The purpose of the study was to explore the relationship between the GPR41 and GPR43 and seasonal breeding, and provide new prospects for further exploring the nutritional needs of breeding. We identified the localization and expression levels of GPR41 and GPR43 in the colon of the wild ground squirrels (Spermophilus dauricus) both in the breeding season and non-breeding season. The histological results revealed that the lumen diameter of the colon had obvious seasonal changes, and the diameter of the colonic lumen in the non-breeding season was larger than that in the breeding season. Immunohistochemical staining suggested GPR41 and GPR43 have expressed in the simple layer columnar epithelium. In addition, compared with the breeding season, the mRNA and protein expression levels of GPR41 and GPR43 in the colon were higher during the non-breeding season. In general, these results indicated GPR41 and GPR43 might play a certain role in regulating seasonal breeding.

Seasonal expression of extracellular signal regulated kinases in the colon of wild ground squirrels (Spermophilus dauricus).

BACKGROUND: The purpose of the experiment was to explore the localization and seasonal expression of extracellular signal regulated kinase (ERK) in the colonic tissue of wild ground squirrels (Spermophilus dauricus). METHODS AND RESULTS: Hematoxylin-eosin staining, immunohistochemistry, real-time quantitative PCR and Western blotting were used in this experiment. The histological results showed that the diameter of the colon lumen enlarged and the number of glandular cells increased in the non-breeding season. It was found in the immunochemical results that both ERK1/2 and pERK1/2 were expressed in the cytoplasm of goblet cells and intestinal epithelial cells, while pERK1/2 was also expressed in the nucleus of them. The immune localization of both was more obvious in the non-breeding season, especially in intestinal epithelial cells. Real-time quantitative PCR and Western blotting showed that ERK1/2 and pERK1/2 were seasonally highly expressed in the non-breeding season. CONCLUSIONS: The expression of ERK1/2 and pERK1/2 was seasonal changes and had significant increases in the non-breeding season. This study revealed that ERK1/2 had potential roles in the colon to the adaptation of seasonal changes in wild ground squirrels.

Seasonal Changes in the Distinct Taxonomy and Function of the Gut Microbiota in the Wild Ground Squirrel (Spermophilus dauricus).

Seasonal breeding is a normal phenomenon in which animals adapt to natural selection and reproduce only in specific seasons. Large studies have reported that the gut microbiota is closely related to reproduction. The purpose of this study was to explore the distinct taxonomy and function of the gut microbiota in the breeding and non-breeding seasons of the wild ground squirrel (Spermophilus dauricus). The 16S rRNA gene sequencing technology was utilized to sequence the gut microbiota of the wild ground squirrel. PICRUSt analysis was also applied to predict the function of the gut microbiota. The results suggested that the main components of the gut microbiota in all samples were Firmicutes (61.8%), Bacteroidetes (32.4%), and Proteobacteria (3.7%). Microbial community composition analyses revealed significant differences between the breeding and non-breeding seasons. At the genus level, Alistipes, Mycoplasma, Anaerotruncus, and Odoribacter were more abundant in the non-breeding season, while Blautia and Streptococcus were more abundant in the breeding season. The results of a functional prediction suggested that the relative abundance of functional categories that were related to lipid metabolism, carbohydrate metabolism, and nucleotide metabolism increased in the breeding season. The relative abundance of energy metabolism, transcription, and signal transduction increased in the non-breeding season. Overall, this study found differences in the taxonomy and function of the gut microbiota of the wild ground squirrel between the breeding and non-breeding seasons, and laid the foundation for further studies on the relationship between the gut microbiota and seasonal breeding.

Population Genetic Structure Analysis Reveals Decreased but Moderate Diversity for the Oriental Fire-Bellied Toad Introduced to Beijing after 90 Years of Independent Evolution.

Detailed molecular genetic research on amphibian populations has a significant role in understanding the genetic adaptability to local environments. The oriental fire-bellied toads (Bombina orientalis) were artificially introduced to Beijing from Shandong Province in 1927, and since then, this separated population went through an independent evolution. To explore the differentiation of the introduced population with its original population, this study analyzed the genetic structure of the oriental fire-bellied toad, based on the mitochondrial genome control region and six microsatellite sites. The results showed that the haplotype diversity and nucleotide diversity of the mitochondrial D-loop region partial sequences of the Beijing Botanical Garden population and the Baiwangshan population were lower than those of the Shangdong Kunyushan population. Microsatellite marker analysis also showed that the observed heterozygosity and expected heterozygosity of the Beijing populations were lower than those of the Kunyushan population. The phylogenetic trees and network diagrams of haplotypes indicated that the three populations were not genetically separated. However, the structure analysis showed a genetic differentiation and categorized the sampling individuals into Beijing and Shandong genetic clusters, which indicated a tendency for isolated evolution in the Beijing population. Although the Beijing populations showed a decline in genetic diversity, it was still at a moderate level, which could maintain the survival of the population. Thus, there is no need to reintroduce new individuals from the Kunyushan source population.

A Study of Population Size and Activity Patterns and Their Relationship to the Prey Species of the Eurasian Lynx Using a Camera Trapping Approach.

Revealing the behavioral relationships between predators and their prey is fundamental in understanding the community structure and ecosystem functions of such animals. This study aimed at detecting the population size and activity patterns of Eurasian lynx (Lynx lynx) (along with its prey) by camera trapping monitoring from 2014 to 2017 at the Saihanwula nature reserve in central Inner Mongolia. The total effective trapping days were 29,892 and 20 lynx were identified from 343 trapping photos based on the inner side patterns of their forelimbs. The daily activity rhythms of the lynx overlapped with those of different prey in different seasons. The yearly activity pattern of the lynx was influenced by its main prey's biology. In conclusion, this study reveals that the activity patterns of the top predator matched those of its prey in different time periods. Habitat management strategies promoting the restoration of prey communities would benefit the lynx in maintaining a stable community structure.

The role of the adiponectin system in acute fasting-impaired mouse ovaries.

Adiponectin (ADIPOQ, encoded by Adipoq) is an important white adipose-derived adipokine linked to energy homeostasis and reproductive function. This study aims to reveal the expression and role of the adiponectin system in the ovaries under acute malnutrition. In this study, 48-h food deprivation significantly inhibited ovarian growth by suppressing cell proliferation and inducing cell apoptosis in the ovaries of gonadotrophin-primed immature mice. It was also accompanied by significantly decelerated basic metabolism (glucose, triacylglycerol and cholesterol), varied steroid hormones (follicle-stimulating hormone, luteinizing hormone and estradiol) and vanishment of the peri-ovarian fat. It is noteworthy that after acute fasting, the adiponectin levels in ovaries rather than in blood were significantly elevated. Immunohistochemical study demonstrated that adiponectin and its receptors (ADIPOR1 and ADIPOR2) primarily appeared in ovarian somatic and/or germ cells, and their protein expressions were upregulated in the ovaries from fasted mice. Further in vitro study verified that ADIPOR1/2 agonist obviously inhibited follicle-stimulating hormone-induced oocyte meiotic resumption, while the antagonist significantly enhanced the percentage of oocyte maturation in the absence of follicle-stimulating hormone. Furthermore, the build up of peri-ovarian fat under physiological status in mice showed a positive correlation with both the hypertrophy of adipocytes and growth of ovaries. Taken together, these findings indicate that the upregulation of the adiponectin system disturbs the normal female reproductive function under the malnutrition status, and it may be associated with the loss of peri-ovarian fat depots.

Non-invasive genetic analysis indicates low population connectivity in vulnerable Chinese gorals: concerns for segregated population management.

Detailed information on the size and genetic structure of wildlife populations is critical for developing effective conservation strategies, especially for those species that have suffered population decline and fragmentation due to anthropogenic activities. In the present study, we used a non-invasive approach combining fecal pellet sampling with mitochondrial DNA and nuclear DNA microsatellite marker analysis to monitor and compare the population structure of the Chinese goral (Naemorhedus griseus) in Beijing and northeast Inner Mongolia in China. Of the 307 fecal samples confirmed to be from N. griseus, 15 individuals (nine females and six males) were found in the Beijing population and 61 individuals (37 females and 24 males) were found in the Inner Mongolian population. Among these 76 individuals, we identified eight haplotypes and 13 nucleotide polymorphic sites from mtDNA and 45 alleles from 10 microsatellite loci. Spatially structured genetic variation and a significant level of genetic differentiation were observed between the two populations. In both populations, the sex ratios were skewed toward females, indicating high reproductive potential, which is crucial for population recovery and conservation of this patchily distributed vulnerable species. We suggest that managing the two populations as evolutionarily significant units with diverse genetic backgrounds could be an effective solution for present population recovery, with the possible relocation of individuals among different groups to help ensure future goral species prosperity.

Seasonal expressions of prolactin, prolactin receptor and STAT5 in the scented glands of the male muskrats (Ondatra zibethicus).

Prolactin (PRL) production in mammals has been demonstrated in extrapituitary gland, which can activate autocrine/paracrine signaling pathways to regulate physiological activity. In the current study, we characterized the gene expression profiles of PRL, prolactin receptor (PRLR) and signal transducers and activators of transcription 5 (STAT5) in the scented glandular tissues of the muskrats, to further elucidate the relationship between PRL and the scented glandular functions of the muskrats. The weight and volume of the scented glands in the breeding season were significantly higher than those of the non-breeding season. Immunohistochemical data showed that PRL, PRLR and STAT5/phospho-STAT5 (pSTAT5) were found in the glandular and epithelial cells of the scented glands in both seasons. Furthermore, we found that PRL, PRLR and STAT5 had higher immunoreactivities in the scented glands during the breeding season when compared to those of the non-breeding season. In parallel, the gene expressions of PRL, PRLR and STAT5 were significantly higher in the scented glands during the breeding season than those of the non-breeding season. The concentrations of PRL in scented glandular tissues and sera were measured by enzyme-linked immunosorbent assay (ELISA), and their levels were both notably higher in the breeding season than those of the non-breeding season. These findings suggested that the scented glands of the muskrats were capable of extrapituitary synthesis of PRL, which might attribute PRL a specific function to an endocrine or autocrine/paracrine mediator.

Seasonal expressions of androgen receptor, P450arom and estrogen receptors in the epididymis of the wild ground squirrel (Citellus dauricus Brandt).

The aim of this study was to investigate the seasonal expressions of androgen receptor (AR), estrogen receptors alpha and beta (ERα and ERß) and aromatase cytochrome P450 (P450arom) in the epididymis of the wild ground squirrel. Histologically, the epididymis was with larger duct diameter and cell population during the breeding season. AR was presented in the peritubular smooth muscle cells and epithelial cells in the whole epididymis with stronger staining in the breeding period. P450arom was intensely localized in epithelial cells and spermatozoa during the breeding season, absent in the non-breeding season and moderately stained in pre-hibernation. During the breeding season, ERα was intensely expressed in epithelial cytoplasm and/or nucleus, whereas in the non-breeding season and pre-hibernation, weaker staining signal was found in nucleus of epithelial cells. ERß was absent in the entire annual cycle by immunohistochemical and Real-time PCR detection. The mRNA levels of AR, P450arom and ERα were higher in the epididymis of the breeding season when compared to those of the non-breeding season and pre-hibernation. Taken together, these results suggest that epididymis of the wild ground squirrel is a primary target for androgen and estrogen, and the expression of P450arom represents that epididymis may be a potential source of estrogen.

Isolation, Purification and Structural Characterization of Two Novel Water-Soluble Polysaccharides from Anredera cordifolia.

Anredera cordifolia, a climber and member of the Basellaceae family, has long been a traditional medicine used for the treatment of hyperglycemia in China. Two water-soluble polysaccharides, ACP1-1 and ACP2-1, were isolated from A. cordifolia seeds by hot water extraction. The two fractions, ACP1-1 and ACP2-1 with molecular weights of 46.78 kDa ± 0.03 and 586.8 kDa ± 0.05, respectively, were purified by chromatography. ACP1-1 contained mannose, glucose, galactose in a molar ratio of 1.08:4.65:1.75, whereas ACP2-1 contained arabinose, ribose, galactose, glucose, mannose in a molar ratio of 0.9:0.4:0.5:1.2:0.9. Based on methylation analysis, ultraviolet and Fourier transform-infrared spectroscopy, and periodate oxidation the main backbone chain of ACP1-1 contained (1→3,6)-galacturonopyranosyl residues interspersed with (1→4)-residues and (1→3)-mannopyranosyl residues. The main backbone chain of ACP2-1 contained (1→3)-galacturonopyranosyl residues interspersed with (1→4)-glucopyranosyl residues.

Alternative splicing affects the targeting sequence of peroxisome proteins in Arabidopsis.

KEY MESSAGE: A systematic analysis of the Arabidopsis genome in combination with localization experiments indicates that alternative splicing affects the peroxisomal targeting sequence of at least 71 genes in Arabidopsis. Peroxisomes are ubiquitous eukaryotic cellular organelles that play a key role in diverse metabolic functions. All peroxisome proteins are encoded by nuclear genes and target to peroxisomes mainly through two types of targeting signals: peroxisomal targeting signal type 1 (PTS1) and PTS2. Alternative splicing (AS) is a process occurring in all eukaryotes by which a single pre-mRNA can generate multiple mRNA variants, often encoding proteins with functional differences. However, the effects of AS on the PTS1 or PTS2 and the targeting of the protein were rarely studied, especially in plants. Here, we systematically analyzed the genome of Arabidopsis, and found that the C-terminal targeting sequence PTS1 of 66 genes and the N-terminal targeting sequence PTS2 of 5 genes are affected by AS. Experimental determination of the targeting of selected protein isoforms further demonstrated that AS at both the 5' and 3' region of a gene can affect the inclusion of PTS2 and PTS1, respectively. This work underscores the importance of AS on the global regulation of peroxisome protein targeting.

Correlation of epigenetic aberrance with STAT3 signaling pathway in gastric carcinogenesis.

BACKGROUND: It has been suggested that STAT3 signaling plays important roles in regulating epigenetic aberrance during tumorigenesis, especially in the expression of certain key epigenetic enzymes such as DNMTs, HDACs, and HMTs. However, there has been no report on the relationship of STAT3 signaling and epigenetic aberrance in gastrocarcinogenesis. AIM: The purpose of this study was to explore the interrelationship of STAT3 signaling pathway and epigenetic aberrance in gastrocarcinogenesis. METHODS: Immunohistochemistry was utilized to examine the protein expressions of pSTAT3, DNMT1, HDAC1, and EZH2 in 153 tissue specimens, including 20 of normal gastric epithelium tissue, 21 of intestinal metaplasia (IM), 24 of dysplasia (DYS), 23 of early gastric cancer (EGC) and 65 of advanced gastric cancer (AGC), and then analyze their possible relationship with clinicopathological factors. RESULTS: We found that the four protein expressions were obviously enhanced following the malignant process of gastric carcinogenesis. Pearson correlation analysis of all the pathological groups showed that expression of pSTAT3 was highly associated with DNMT1, but not with HADC1 and EZH2. However, significant correlations were detected among the expression of DNMT1, HDAC1, and EZH2. Further analysis of each pathological group demonstrated that pSTAT3's expression was dramatically related with DNTM1 in the IM (P = 0.021) and EGC groups (P = 0.013) and correlated with EZH2 in the DYS group (P = 0.020). Furthermore, pSTAT3's expression was associated with T staging (P = 0.015) in the AGC group, whereas DNMT1 was associated with gender (P = 0.021), HDAC1 with Lauren classification (P = 0.007), and EZH2 with T staging (P = 0.003) and lymphatic staging (P = 0.038). CONCLUSIONS: The STAT3 signaling pathway may correlate with epigenetic aberrance during gastrocarcinogenesis.