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The purpose of this study was to investigate the relationship between the expression of autophagy-related genes and prognosis in hepatocellular carcinoma (HCC).We selected three autophagy-related genes (ATG3, ATG7, and ATG9A) from gene expression data of liver cancer patients in The Cancer Genome Atlas (TCGA) database by Kaplan-Meier survival analysis, univariate and multivariate Cox regression analysis, and Gene Set Enrichment Analysis (GSEA). Human Protein Atlas (HPA) and Gene Expression Profiling Interactive Analysis (GEPIA) databases were applied to testify the credibility of our results. The expression levels of ATG3, ATG7, and ATG9A were verified by real-time quantitative PCR (RT-qPCR) in normal liver cells (L02) and three HCC cell lines (HepG2, Hep3b, and Li-7). Data analysis results from TCGA showed high ATG3, ATG7, ATG9A expression in HCC tumor tissues. Kaplan-Meier survival analysis showed that the survival rate of the high expression group of ATG3, ATG7, and ATG9A was all significantly lower than the low expression group. GSEA analysis showed that many signaling pathways (such as the regulation of autophagy, glycine serine and threonine metabolism, pathways in cancer, mitogen-activated protein kinase (MAPK) signaling pathway, mammalian target of rapamycin (mTOR) signaling pathway, as well as P53 signaling pathway) were differentially enriched in HCCs with ATG3, ATG7, and ATG9A expression. GEPIA and RT-qPCR also identified that the mRNA expression level of ATG3, ATG7, and ATG9A in normal liver cells were significantly lower than in HCC cells. High protein expression of ATG3, ATG7, and ATG9A was displayed in HCCs from the HPA database.The ATG3, ATG7, ATG9A might be utilized as prognostic biomarkers for liver cancer.

Qinghai-Tibetan Plateau is an important ecological security barrier in China, and alpine ecosystem presents a trend of overall improvement under the influence of climate change and human activities, yet there are localized deteriorations. In order to improve the ecological function of the Tibetan Plateau, large-scale ecological restoration projects (ERPs) have been carried out in Tibetan Plateau over the past 30 years. From following the footstep of national ecological constructions to the implementation of projects specific to the plateau characteristics, the ERP of the plateau can be divided into three stages. Major ERPs focusing on four types of projects, i.e. forest protection and construction, grassland protection and construction, water and soil erosion control and desertification land management, with a total area of 850,000km across the Qinghai-Tibetan Plateau. The positive effect at sampling quadrat scale has been widely verified, and the responses of productivity and species diversity were inconsistent. The positive effect at the regional scale gradually emerges, yet the spatial difference is significant, and the quantification of driving forces is an important prerequisite. The positive and negative effects of existing ERP on biodiversity conservation are revealed, for instance, there is a significant increase of rare wild animals in natural reserves and in artificially planted areas it failed to effectively curb the loss of animal and plant diversity. The long-term effects of various ERPs on the biodiversity should be taken into account so as to optimize the ecological measures and build a sustainable management model after project restoration. By summarizing the achievements and problems of ERP across the Qinghai-Tibetan Plateau in the past 30 years, we proposed to set stepwise ecological restoration and management scheme by different restoration goals and reference modes in three stages, i.e. “Environmental treatment—Ecological restoration projects—Adaptive management”.

The purpose of the present paper is to establish certain fractional integral inclusions having exponential kernels, which are related to the Hermite–Hadamard, Hermite–Hadamard–Fejer, and Pachpatte type inequalities. These results allow us to obtain a new class of inclusions which can be viewed as some substantial generalizations of the previously reported results. Also, the graphical representations for the results are utilized to identify the correctness of the investigated inclusion relations that occur with the change of the parameter $$\alpha $$ .

Mitochondria are the main organelles for mammalian energy metabolism and have been implicated in the regulation of germ cell maintenance and spermatogenesis. However, little is known about the changes in the mitochondria of the testis of seasonal breeders. Here, we characterized the seasonal changes in the mitochondria in the testis of the wild ground squirrels. Increased testicle weight, seminiferous tubule diameter, and sperm count were observed in the wild ground squirrels at the breeding season. RNA-seq analysis of the wild ground squirrel testes revealed that mitochondrial-related genes were expressed differentially between the breeding and nonbreeding seasons. Immunohistochemical staining showed that key mitophagy factors including PINK1, MFN2, and PARKIN were highly expressed in various cell types of testis during the breeding season. In addition, the abundance and enzymatic activities of mitochondrial-localized antioxidative enzymes superoxide dismutase 2 (SOD2) and Catalase were decreased in the testis during the breeding season, suggesting a tightly controlled redox balance at least partially facilitated by mitophagy during the seasonal breeding. Taken together, our study reveals that mitochondrial autophagy and oxidative stress may be implicated in the seasonal reproductive recrudescence of the wild ground squirrels, which deepens our understanding of the mitochondrial regulation of seasonal reproductivity in wildlife and provides new insights into the development of potential therapeutic interventions of male infertility.

The purpose of this study was to explore the variations in the circulating leptin concentrations of the wild ground squirrels in relation to seasonal changes in testicular activities. Hematoxylin-eosin staining showed all types of elongated spermatids and spermatogenic cells existed in the testis in April, while the primary spermatocytes and spermatogonia were most advanced stages of germ cells in June. In addition, the primary spermatocytes, secondary spermatocytes, and spermatogonia were most advanced stages of germ cells in September. The highest circulating leptin concentration was consistent with the maximum body weight results from accumulation of adipose tissue in September. The mRNA expression level of leptin receptor (Ob-R) and STAT3 was lowest in June, raised in September, and remained increased in April. Ob-R and STAT3 were stronger staining in the Leydig cells in July. Moreover, the concentrations of testosterone (T) showed the maximum values in April, the minimum values in June, and significant increases in September. Furthermore, it is worth noting that the levels of T increased with the mRNA levels of Ob-R, STAT3, StAR, and testicular steroidogenic enzymes (3β-HSD, P450c17, and P450scc). Moreover, RNA-seq analyses of testis during the different periods showed that a total of 4209 genes were differentially expressed genes (DEGs); further analysis revealed that DEGs related with the Jak/STAT pathways and reproduction were altered. Taken together, the results suggested that the leptin regulated testicular function through the Jak/STAT pathways and testicular steroidogenic factor expressions.

We utilize the definition of a fractional integral operators, which was presented by Ahmad et al., to investigate a general fractional-type identity with a parameter. We establish certain parameterized fractional integral inequalities based on this identity, and provide two examples to illustrate the obtained results. Also, these results derived in this paper are applied to the estimations of q-digamma function, divergence measures and cumulative distribution function, respectively.

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.

Mitochondria are the main organelles for mammalian energy metabolism and have been implicated in the regulation of germ cell maintenance and spermatogenesis. However, little is known about the changes in the mitochondria of the testis of seasonal breeders. Here, we characterized the seasonal changes in the mitochondria in the testis of the wild ground squirrels. Increased testicle weight, seminiferous tubule diameter, and sperm count were observed in the wild ground squirrels at the breeding season. RNA-seq analysis of the wild ground squirrel testes revealed that mitochondrial-related genes were expressed differentially between the breeding and nonbreeding seasons. Immunohistochemical staining showed that key mitophagy factors including PINK1, MFN2, and PARKIN were highly expressed in various cell types of testis during the breeding season. In addition, the abundance and enzymatic activities of mitochondrial-localized antioxidative enzymes superoxide dismutase 2 (SOD2) and Catalase were decreased in the testis during the breeding season, suggesting a tightly controlled redox balance at least partially facilitated by mitophagy during the seasonal breeding. Taken together, our study reveals that mitochondrial autophagy and oxidative stress may be implicated in the seasonal reproductive recrudescence of the wild ground squirrels, which deepens our understanding of the mitochondrial regulation of seasonal reproductivity in wildlife and provides new insights into the development of potential therapeutic interventions of male infertility.

Simple Summary Gut microbiota is a large number of microbes colonized in the gut tract, and it plays a certain role in regulating the host’s immunity, metabolism, and nervous system. Recent studies have shown that the gut microbiota also has a close relationship with reproduction. The wild ground squirrel (Spermophilus dauricus) is a typical seasonal breeding animal. 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 using 16S rRNA gene sequencing technology. The results show that the taxonomy of gut microbiota was different between the breeding season and non-breeding season. Functional prediction of the gut microbiota indicated that the relative abundance of metabolic pathways was differentially enriched between the breeding season and non-breeding season. This study further revealed the potential relationship between gut microbiota and reproduction and expanded our understanding of the function of gut microbiota. At the same time, it provided a new direction for research on the breeding strategy of seasonal breeding animals. Abstract 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.

Background: The aim of this study was to explore the localization and expression of extracellular signal regulated kinase/phospho-extracellular signal regulated kinases (ERK/pERK) in the colonic tissue of wild ground squirrels (Spermophilus dauricus) in the breeding season and the non-breeding season. Methods and Results: Hematoxylin-eosin staining, immunohistochemistry, real-time quantitative PCR and Western blot were used in this study. The histological results showed that the diameter of the colon lumen in the non-breeding season was larger than that in the breeding season, and the number of glandular cells in the non-breeding season was also more than those in the breeding season. The immunochemical results showed that ERK1/2 was expressed in the cytoplasm of goblet cells and intestinal epithelial cells, while pERK1/2 was expressed in the cytoplasm and nucleus of goblet cells and intestinal epithelial cells. The immunolocalization of ERK1/2 and pERK1/2 expression were more obvious in the non-breeding season, especially in intestinal epithelial cells. The results of real-time quantitative PCR and Western blot showed that the expression of ERK1/2 and pERK1/2 in the breeding season was significantly lower than that in the non-breeding season.Conclusions: The expression of ERK1/2 and pERK1/2 in the colons of the wild ground squirrels had seasonal changes, which had significant increases in the non-breeding season comparing to those in the breeding season. This study revealed the potential role of ERK1/2 in colon to the adaptation of seasonal changes in wild ground squirrel.

3-Methyl-4-Nitrophenol (PNMC) is the main degradation product of organophosphate insecticide fenitrothion and a major component of diesel exhaust particles, which is now becoming a widely spread environmental endocrine disruptor. Previous reports showed PNMC exposure can affect the female reproductive system and ovarian function; however, the mechanism remains unclear. The main purpose of this study is to clarify the mechanism underlying the adverse effects of neonatal PNMC treatment on ovarian functions. The neonatal female mice were exposed to 10 mg/kg PNMC and the ovaries were collected on the 7th day after birth. The changes of follicular composition in mice ovaries were analyzed by histological staining, which showed that the proportion of primordial follicles in the ovaries treated by PNMC decreased, while the proportion of secondary follicles increased. The ovarian function was also investigated by detecting the expressions of steroidogenic enzymes (Star, Cyp11a1, Hsd3b1, Cyp17a1, Cyp19a1), gonadotropin receptors (Fshr and Lhr), androgen receptor (Ar), and estrogen receptors (Esr1 and Esr2) by immunohistochemistry or/and real-time quantitative PCR. The expression of Hsd3b1, Cyp17a1 and Esr2 were increased significantly in the PNMC exposed ovaries. Moreover, the expression patterns of clock genes (Bmal1, Clock, Per1, Per2, Cry1, Cry2 and Nr1d1) were disrupted in the ovaries after PNMC exposure. Furthermore, either the expression of DNA Methyltransferase Dnmt3b, or the methylation ratio of CpG islands in the upstream of Cry1 promoter regions were significantly decreased in PNMC exposed ovaries. Altogether, these results indicate that PNMC exposure affects follicle development and ovarian function by interfering with the epigenetic modification and disrupting the expression of clock genes.

Epidermal growth factor (EGF) is an important autocrine and/or paracrine mediator of steroid hormones to stimulate growth and differentiation in mammals. The aim of this study is to investigate seasonal expressions of estrogen receptor α (ERα), estrogen receptor β (ERβ), EGF, epidermal growth factor receptor (EGFR), phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt) in the scent glands of the muskrats during the breeding and non-breeding seasons. Histologically, three types of cells including the glandular cells, interstitial cells and epithelial cells were identified in the scent glands in both seasons. Immunohistochemical results showed that ERα, ERβ, EGF, EGFR, PI3K and Akt were presented in the different types of cells of the scent glands during the breeding and non-breeding seasons. Transcriptome data of the scent glandular tissues from muskrats in the breeding and non-breeding seasons showed that differential seasonal changes might be related to the estrogen-EGFR signaling pathway. The gene expression levels of ERα, ERβ, EGF, EGFR, PI3K were increased, while the gene expression level of Akt were decreased in the breeding season than those in the non-breeding season. Besides, the concentrations of 17β-estradiol (E2) in the serum and the scent glandular tissues were remarkably higher in the breeding season than those of the non-breeding season. Taken together, our results suggested that EGFR signaling pathway may coordinate with ERs signaling to regulate the seasonal changes of the scent glandular functions.

As a vital male accessory reproductive gonad, the prostate requires vascular endothelial growth factors for promoting its growth and development. In this study, we investigated the localizations and expressions of vascular endothelial growth factor (VEGF) and its receptors including VEGF-receptor1 (VEFGR1) and VEGF-receptor2 (VEGFR2) in the prostate of the wild ground squirrels during the breeding and the non-breeding seasons. The values of total prostate weight and volume in the breeding season were higher than those in the non-breeding season. Histological observations showed that the exocrine lumens of the prostate expanded in the breeding season and contracted in the non-breeding season. The mRNA expression levels of VEGF and VEGFR2 in the prostate were higher in the breeding season than those in the non-breeding season, but the mRNA expression level of VEGFR1 had no significant change between the breeding and non-breeding seasons. Immunohistochemical results revealed that VEGF, VEGFR1 and VEGFR2 were presented in epithelial and stromal cells during the breeding and non-breeding seasons. In addition, the microvessels of the prostate were widely distributed and the number of microvessels increased obviously in the breeding season, while decreased sharply in the non-breeding season. These results suggested that expression levels of VEGF and VEGFR2 might be correlated with seasonal changes in morphology and functions of the prostate, and VEGF might serve as pivotal regulators to affect seasonal changes in the prostate functions of the wild male ground squirrels via an autocrine/paracrine pathway.

Previous studies found that testosterone was converted to dihydrotestosterone under the catalysis of 5α-reductase in the prostate of the wild ground squirrels. As a result, this study explored further whether testosterone could be converted to estrogen to affect the prostate gland function in wild ground squirrels. Histological observation showed that the area of epithelial cells and the prostatic secretory lumen were enlarged significantly during the breeding period. Transcriptome analysis revealed that the differentially expressed genes in the prostate were concentrated in the estrogen signaling pathway. Immunohistochemical analysis showed that the immunoreactivities of P450arom were detected in the stromal cells during the breeding and non-breeding periods, indicating the possible conversion of androgen into estrogen locally. Moreover, the immunolocalizations of ERα and ERβ were detected mainly in the epithelial or stromal cells. Additionally, qPCR analysis displayed that the mRNA expression level of P450arom in the prostate was significantly higher during the breeding period than that in the non-breeding period. Consistently, the concentration of 17β-estradiol (E

The prostate gland is a male accessory reproductive gland that requires androgenic steroids for maintaining its vitality and function. The aim of the study was to investigate the localization and expressions of the androgen receptor (AR), SRD5A1 and SRD5A2 in the prostate glands of wild male ground squirrels during different reproductive periods by immunohistochemistry and real-time PCR. Gross mass of the prostate gland was higher in the breeding season than that of the non-breeding season. Histological observation showed that exocrine lumens in the prostate gland were enlarged in the breeding season and shrank in the non-breeding season. Immunohistochemical results demonstrated that AR was presented in both epithelial and stromal cells' nuclei during both the breeding and non-breeding seasons. Intriguingly, the positive staining of SRD5A1 and SRD5A2 was only found in the stromal cells. The mRNA expression of the three genes including Ar, Srd5a1 and Srd5a2 in the prostate gland was higher in the breeding season than those of the nonbreeding season. In addition, the circulating concentration of testosterone (T) and the concentration of dihydrotestosterone (DHT) in the prostate gland were also markedly higher in the breeding season than those of the nonbreeding season. These results suggest that expression levels of AR, SRD5A1 and SRD5A2, as well as DHT synthesis, might be correlated with seasonal changes in morphology and function of the prostate gland, indicating that DHT may serve as a pivotal regulator to affect the morphology and function of prostate gland via a autocrine/paracrine pathway in wild male ground squirrels.

Background: Seasonal breeding is a normal phenomenon that animals adapt to natural selection and reproduce only in specific seasons. With the gradual popularization of Next-generation sequencing (NGS), large studies have shown that seasonal breeding has been affected by gut microbiota. Consequently, the purpose of this study is to explore the effect of seasonal breeding on the gut microbiota of wild ground squirrel (Spermophilus dauricus). We used 16S rRNA gene sequencing technology to sequence the gut microbiota of the wild ground squirrel in the breeding season and non-breeding season. We also predicted the function of gut microbiota by bioinformatic software.Results: The results showed that the main components of gut microbiota in all samples consisted of Firmicutes (61.8%), Bacteroidetes (32.4%), and Proteobacteria (3.7%). Microbial community composition analyses revealed significant differences between these two groups. At the genus level, Alistipes, Mycoplasma, Anaerotruncus, and Odoribacter were up-regulated in the non-breeding season, while Blautia and Streptococcus spp. were up-regulated in the breeding season. The result of function prediction suggested that the relative abundance of functional categories related to lipid metabolism, carbohydrate metabolism, and nucleotide metabolism was higher in the breeding season. The expression of transcription, energy metabolism, and signal transduction was enriched in the non-breeding season. Conclusions: Overall, the results of this study emphasized the significant effects of seasonal breeding on gut microbiota community composition of the wild ground squirrel and laid a foundation for further study of gut microbiota on seasonal breeding in the future.

The resale right system, first created in France in 1920, has a history of one hundred years and has been confirmed by dozens of national legislations. In general, the introduction of the resale right has become a trend in the legislation of various countries. But the universal establishment of the resale right does not necessarily mean that it has legitimacy. On the issue of whether the introduction of the resale right is justified, the views of supporters and opponents are tit-for-tat, and several major points of controversy have formed. By sorting out the disputes involved in the resale right system, clarifying the core problems of introducing the resale right, and demonstrating its legitimacy, it is reasonable and feasible to introduce the resale right.

Duchenne Muscular Dystrophy (DMD) is caused by mutation of the muscle membrane protein dystrophin and characterized by severe degeneration of myofibers, progressive muscle wasting and loss of mobility, ultimately cardiorespiratory failure and premature death. Here we report that skeletal muscle-specific knockout (KO) of Phosphatase and tensin homolog (Pten) gene in an animal model of DMD (mdx mice) alleviates myofiber degeneration and restores muscle function without increasing tumor incidences. Specifically, Pten KO normalizes myofiber size and prevents muscular atrophy, and improves grip strength and exercise performance of mdx mice. Pten KO also reduces fibrosis and inflammation; and ameliorates muscle pathology in mdx mice. Moreover, we found that Pten KO upregulates extracellular matrix and basement membrane components positively correlated to wound healing, but suppresses negative regulators of wound healing and lipid biosynthesis; and restores the integrity of muscle basement membrane in mdx mice. Importantly, pharmacological inhibition of PTEN similarly ameliorates muscle pathology and improves muscle integrity and function in mdx mice. Our finding provides evidence that PTEN inhibition may represent a potential therapeutic strategy to restore muscle function in DMD.

We use the definition of a new class of fractional integral operators, recently introduced by Ahmad et al. in [J. Comput. Appl. Math. 353:120–129, 2019], to establish a fractional-type integral identity with one parameter. We derive some parameterized integral inequalities for convex mappings based on this identity, and provide two examples to illustrate the investigated results as well. Moreover, we present applications of our findings to special means of real numbers, and error estimations for the quadrature formula in numerical analysis.

The seasonal cycle of growth and regression in the prostate gland of wild ground squirrel provide a unique research model to understand the morphological changes of prostate glands. Our previous studies showed that the local production of dihydrotestosterone could affect the morphology and function of the prostate gland in either an autocrine or paracrine manner. In the present study, we attempted to gain more insight into this process by investigating the expression of key factors implicated in cell proliferation, apoptosis, and the cell cycle, including mechanistic target of rapamycin (mTOR), cyclin-D2, p21, p27 and retinoblastoma 1 (pRB). Morphological and histological observations confirmed that the prostate increased significantly in both size and weight during the breeding season. Positive immunostaining for proliferating cell nuclear antigen (PCNA) was mainly localized to the prostate epithelial cells during the breeding season, which is significantly higher in the prostate gland during the breeding season (2470 ± 81/mm2) than that in the nonbreeding season (324 ± 54/mm2). However, there was no significant difference in the prostate gland when compared between the breeding and nonbreeding seasons, with regards to TUNEL staining. Moreover, cell cycle regulators were mainly localized to the epithelial cells, including mTOR, cyclin-D2, p21, p27 and pRB. the immunostaining of mTOR and cyclin D2 were stronger during the breeding season, whereas the immunostaining of p27 and pRB were stronger during the nonbreeding season. The mRNA expression levels of mTOR, cyclin D2, and PCNA, were higher during the breeding season while those of p27 and p21 were higher during the nonbreeding season. Collectively, this study profiled the distinct expression pattern of key cell cycle regulators throughout the breeding and nonbreeding seasons. Collectively, these factors may play important roles in regulating the seasonal growth and regression of the prostatic epithelium in the wild ground squirrel.

Sperm-associated antigen 11A (SPAG11A), a protein expressed exclusively in the epididymis, plays a vital and special role in regulating mammalian sperm maturation. The aim of this study was to investigate the seasonal expressions of SPAG11A and androgen receptor (AR) in the epididymis of the wild ground squirrels (Spermophilus dauricus Brandt). Morphologically, the results showed that epididymis length and weight in the breeding season were significantly higher than those of the non-breeding season. Histologically, the results revealed that enlarged lumen diameters, thickened epithelium and abundant sperm in the breeding season while reduced lumen diameters and epithelium with no sperm in the non-breeding season. SPAG11A was intensely expressed in cytoplasm and nucleus of epithelial cells and smooth muscle cells in the breeding season, and weaker staining in the non-breeding season. In the lumen of epididymis, SPAG11A immunostaining in the sperm of the epididymal corpus and cauda was higher than those in the caput during the breeding season. The immunostaining of AR was only presented in nucleus of smooth muscle cells and epithelial cells in the epididymis from the breeding season rather than the non-breeding season. The results of real-time quantitative PCR also showed that the mRNA levels of SPAG11A and AR in the epididymis during the breeding season were significantly higher than those of the non-breeding season. In addition, the circulating testosterone, follicle- stimulating hormone and luteinizing hormone levels in the squirrels were higher in the breeding season compared with those in the non-breeding season. Taken together, these results implied that SPAG11A might be involved in regulating seasonal changes in epididymal functions of the wild ground squirrels.

This study aimed to investigate the impact of TP53 alteration on survival and clinicopathological features of glioma patients with H3K27M mutations. An individual-participant-data (IPD) meta-analysis was performed to investigate the impact of TP53 alteration on survival and clinicopathological features of patients with H3K27M mutations. Three hundred thirty-one individual records from 12 eligible glioma studies involving the H3K27M mutation were finally included in our meta-analysis, and a pooled hazard ratio (HR) of 1.53 (95%CI, 1.10–2.11; P = 0.01) indicated that TP53 alterations were associated with a shorter overall survival. The pooled odds ratios (ORs) indicated that TP53 alterations were significantly associated with the age at diagnosis ≥ 7 years (OR = 1.97, 95%CI = 1.15–3.38, P = 0.01), the status of histone H3.3 mutations (OR = 9.15, 95%CI = 4.18–20.06, P

Oxytocin (OT) can modulate multiple physiological functions via binding to the widely distributed oxytocin receptor (OTR). In this study, we investigated the seasonal expressions of OT, OTR and extracellular signal regulated kinase (ERK1/2) signaling pathway components in the scented gland of muskrat during the breeding and non-breeding seasons. Histologically, glandular cells, interstitial cells and excretory tubules were identified in the breeding season scented glands, whereas epithelial cells were sparse in the non-breeding season. Immunohistochemical results showed that OTR was present in epithelial cells and interstitial cells while OT, pERK1/2, ERK1/2 and c-fos were expressed in epithelial cells and glandular cells. The protein and mRNA expressions of OTR, OT and c-fos were significantly higher in the scented gland in the breeding season than in the non-breeding season. Importantly, the levels of OT in scented glands and serum were measured by hormone assays, and their concentrations were both significantly higher in the breeding season than in the non-breeding season. Moreover, bioinformatics analysis showed that the predicted targets of the differentially expressed microRNAs might include the genes encoding OTR, ERK1/2 and c-fos. These findings suggested that OT may regulate the function of muskrat scented glands by the locally expressed receptors.

Male muskrats have one pair of scented glands that grow and involute annually. To investigate the annual changes in the scented gland, we measured the expressions of aromatase cytochrome P-450 (P450arom) and estrogen receptors (ERs) in the scented glands. P450arom was expressed in glandular cells and epithelial cells in the scented glands during the breeding season, and only in glandular cells during the nonbreeding season. ERα and ERβ were also detected in different types of cells in the scented gland during the breeding and nonbreeding seasons. Both mRNA and protein levels of P450arom, ERα, and ERβ were higher in the scented glandular tissues during the breeding season than those during the nonbreeding season. In addition, small RNA sequencing showed that the predicted targets of the significantly changed microRNAs might be the genes encoding P450arom and ERs. In conclusion, the seasonal changes in the expression of P450arom and ERs may be involved in the regulation of scented gland functions.

The cytokine interleukin-1 beta (IL-1β) is an evolutionarily conserved molecule that was originally identified in the immune system. Nuclear factor κB (NF-κB) plays a critical role in the activation of immune cells by upregulating the expression of many cytokines. In this study, we investigated the localization and expression level of IL-1β, its functional membrane receptor type I (IL-1R1) and NF-κB in the skin of Rana dybowskii during the breeding period and pre-hibernation. Histologically, the skin of Rana dybowskii consists of epidermis and dermis, and four kinds of cells were identified in the epidermis during the breeding period and pre-hibernation, while the dermis was composed of homogenous gel, mucous glands and granular glands. IL-1β, IL-1R1 and NF-κB were immunolocalized in the epithelial and glandular cells in both periods. Western blotting showed that there was no significant difference in the expression of IL-1β between the breeding period and pre-hibernation, whereas IL-1R1 and NF-κB were significantly higher in the pre-hibernation compared to the breeding period. These results suggested that IL-1β and NF-κB may collectively play important roles in the skin immune system of Rana dybowskii during the breeding period and pre-hibernation.

The prostate gland is a male accessory reproductive gland, whose vitality and function are under tight regulation of different hormones. Prostaglandins E2 (PGE2) is one of the major products generated by the actions of cyclooxygenases (COX) and prostaglandin E synthase (PTGES) on arachidonic acid, and is involved in a number of physiological and pathological processes. In this study, we investigated the seasonal immunolocalizations and expressions of COX-1, COX-2 and PTGES, as well as PGE2 receptors (PTGERs) subtypes 1-4 (EP1, EP2, EP3, EP4) in the prostate of the wild ground squirrel. Histological examination observed enlarged prostatic lumens in the breeding season and significantly shrunken lumens in the nonbreeding season. COX-1, COX-2, PTGES and PTGERs were mainly localized in epithelial and stromal cells in the breeding and nonbreeding seasons. The mRNA expression levels of Cox-1, Cox-2, Ptges, Ptger2 (encoding EP2) and Ptger4 (encoding EP4) were higher in the prostate of the breeding season than in the nonbreeding season. The relative mRNA levels of Cox-1, Cox-2, Ptges, Ptger2 and Ptger4 were positively correlated with prostatic weights. In addition, both the prostatic and plasma concentrations of PGE2 were significantly higher in the breeding season compared to the nonbreeding season. These results suggested that PGE2 synthesis and signaling might play an important autocrine or paracrine role in the regulation of seasonal changes in the prostatic function of the wild ground squirrel.

The growth hormone (GH)/insulin-like growth factor-1 (IGF-1) system plays an important role in regulating the cellular growth and organ development. The present study investigated the seasonal expressions of growth hormone receptor (GHR), IGF-1 and insulin-like growth factor 1 receptor (IGF-1R) in the scented glands of the muskrats. Morphological changes in the scented glands of the muskrats were observed significantly between the breeding and non-breeding seasons. Immunohistochemically, the expressions of GH, GHR, IGF-1 and IGF-1R were found in glandular cells and epithelial cells of the scented glands in both seasons. The protein and mRNA expression levels of GHR, IGF-1 and IGF-1R in the scented glands during the breeding season were noticeably higher than those of the non-breeding season. In parallel, the levels of GH and IGF-1 in the sera and scented glands were remarkably higher during the breeding season. In addition, small RNA sequencing showed that the predicted targets of the significantly changed hsa-miR-5100 and mmu-miR-6937-5p might regulate the expressions of Ghr, Igf-1 or Igf-1r. These results suggested that the morphological changes in the scented glands of the muskrats during the different seasons might be related to the expression levels of GHR, IGF-1 and IGF-1R. Meanwhile, GHR/IGF-1 system might regulate the scented glandular functions via endocrine or autocrine/paracrine manners.