The precise pathogenic mechanism in Cu exposure-cause nephrotoxicity remains unclear. This study investigated the underlying molecular mechanism of copper sulfate (CuSO₄)-induced nephrotoxicity. Mice were treated with CuSO₄ at 50, 100, 200 mg/kg/day or co-treated with CuSO₄ (200 mg/kg/day) and 4-phenylbutyric acid (4-PBA, 100 mg/kg/day) for 28 consecutive days. HEK293 cells were treated with CuSO₄ (400 μM) with or without superoxide dismutase, catalase or 4-PBA for 24 h. Results showed that CuSO₄ exposure can cause renal dysfunction and tubular necrosis in the kidney tissues of mice. CuSO₄ exposure up-regulated the activities and mRNA expression of caspases-9 and -3 as well as the expression of glucose-regulated protein 78 (GRP78), GRP94, DNA damage-inducible gene 153 (GADD153/CHOP), caspase-12 mRNAs in the kidney tissues. Furthermore, superoxide dismutase and catalase pre-treatments partly inhibited CuSO₄-induced cytotoxicity by decreasing reactive oxygen species (ROS) production, activities of caspases-9 and -3 and DNA fragmentations in HEK293 cells. 4-PBA co-treatment significantly improved CuSO₄-induced cytotoxicity in HEK293 cells and inhibited CuSO₄ exposure-induced renal dysfunction and pathology damage in the kidney tissues. In conclusion, our results reveal that oxidative stress and endoplasmic reticulum stress contribute to CuSO₄-induced nephrotoxicity. Our study highlights that targeting endoplasmic reticulum and oxidative stress may offer an approach for Cu overload-caused nephrotoxicity.

The gut microbiota has key physiological functions in host adaptation, although little is known about the seasonal changes in the composition and diversity of the gut microbiota in deer. In this study, seasonal variations (grassy and withering season) in the gut microbiota of white-lipped deer (Cervus albirostris), which lives in alpine environments, were explored through 16S rRNA high-throughput sequencing based on sixteen fecal samples collected from Gansu Qilian Mountain National Nature Reserve in China. At the phylum level, Firmicutes, Bacteroidota, and Actinobacteriota dominated the grassy season, while Firmicutes, Proteobacteria, and Actinobacteriota dominated the withering season. At the genus level, Carnobacterium dominated the grassy season, while Arthrobacter and Acinetobacter dominated the withering season. Alpha diversity results (Shannon: P = 0.01, ACE: P = 0.00, Chao1: P = 0.00) indicated that there was a difference in the diversity and richness of the gut microbiota between the two seasons, with higher diversity in the grassy season than in the withering season. Beta diversity results further indicated that there was a significant difference in the community structure between the two seasons (P = 0.001). In summary, the composition, diversity, and community structure of the gut microbiota showed significant seasonal variations, which could be explained by variations in the seasonal food availability, composition, diversity, and nutrition due to phenological alternations. The results of this study indicate that the gut microbiota can adapt to changes in the environment and provide the scientific basis for health assessment of white-lipped deer.

Sexual dimorphism (SD) is a widespread phenomenon in most vertebrate species and is exhibited in a myriad of ways. In amphibians, sexual size dimorphism, in which females are larger than males, is the most common type, and sexual shape dimorphism varies among species. Different selection forces (sexual selection, fecundity selection, and ecological selection) that act differently upon the sexes form the consequence of SD. Thus, studies of SD provide information about the general intersexual divergence of the same species and allow insights into the impact of selective forces on the sexes. In this study, we analyzed morphometric data of the Shangcheng stout salamander,Pachyhynobius shangchengensis, an endemic and poorly known Chinese salamander, to examine sexual dimorphism in size and shape. The morphometric data included 15 characteristics of 68 females and 55 males which were analyzed using univariate and multivariate methods. A significant difference was found between the sexes in terms of both body size (snout-vent length) and some body shapes (e.g., head length and width, tail length and width, distance between limbs, and limb length and width) in this salamander. The longer snout-vent length in males may be attributed to sexual selection, longer and wider head in males may contribute to male-male competition, longer and wider tail in males may be attributed to energy storage and reproductive success, the larger distance between limbs in females is likely due to a fecundity advantage, and longer and more robust limbs in males may be related to reproductive or competitive behaviors. These results demonstrated that sexual dimorphism of different morphological traits is the consequence of different selection forces that act differently upon the sexes.

The Yenyuan stream salamander (Batrachuperus yenyuanensis) has been previously evaluated with regards to phylogeny, population genetics, and hematology, but genomic information is sparse due to the giant genome size of salamanders which contain highly repetitive sequences, thus resulting in the lack of a complete reference genome. This study evaluates the encoding genetic sequences and provides the first transcriptome assembly of Yenyuan stream salamander based on mixed samples from the liver, spermary, muscle and spleen tissues. Using this transcriptome assembly and available encoding sequences from other vertebrates, the gene families, phylogenetic status, and species divergence time were compared or estimated. A total of 13,750 encoding sequences were successfully obtained from the transcriptome assembly of Yenyuan stream salamander, estimated to contain 40.1% of the unigenes represented in tetrapod databases. A total of 88.79% of these genes could be annotated to a biological function by current databases. Through gene family clustering, we found multiple possible isoforms of the Scribble gene—whose function is related to regeneration—based on sequence similarity. Meanwhile, we constructed a robust phylogenetic tree based on 56 single-copy orthologues, which indicates that based on phylogenetic position, the Yenyuan stream salamander presents the closest relationship with the Chinese giant salamander (Andrias davidianus) of the investigated vertebrates. Based on the fossil-calibrated phylogeny, we estimated that the lineage divergence between the ancestral Yenyuan stream salamander and the Chinese giant salamander may have occurred during the Cretaceous period (~78.4 million years ago). In conclusion, this study not only provides a candidate gene that is valuable for exploring the remarkable capacity of regeneration in the future, but also gives an interesting insight into the understanding of Yenyuan stream salamander by this first transcriptome assembly.