Rana Chensinensis Research Articles

Cogrowth advantage: Intestinal microbiota analysis of Bufo gargarizans and Rana chensinensis

Intestinal microbiota has profound effects on host health and adaptation to environmental changes. Bufo gargarizans and Rana chensinensis coexist in the same habitat and have been paid much attention to economically because of their medicinal value. To date, no comparison of differences between single and mixed populations has been made. In our study, differences in the structure and function of the intestinal microbial of B. gargarizans and R. chensinensis in environments of single-species and mixed-species growth were investigated by high-throughput sequencing. Our results suggest that the cogrowth of B. gargarizans and R. chensinensis could lead to the decrease of the abundance of pathogenic bacteria (Bosea) and the introduction or increase of beneficial bacteria (Kaistia, Cetobacterium and Erysipelatoclostridium). The Tax4Fun-based functional predictions revealed that the level of pathways involved in the metabolism of R. chensinensis in mixed-species aquaria is greatly up-regulated. This study provides useful information for ecologists, ecosystem policy makers and wildlife conservationists to promote more effective conservation measures.

Effects of natural and mechanical drying processes on unsaturated fatty acids, vitamin E, estradiol, and physicochemical properties of Rana chensinensis ovum

Although Rana chensinensis Ovum (RCO) is enriched with high nutritional value, its quality varies due to the different drying methods used by farmers. This study aims to explore the impact of natural and mechanical drying methods on the nutritional compositions and physicochemical properties of RCO. Five different drying methods were studied, including blast drying (BD), mild breeze light drying (MBLD), natural air drying (NAD), vacuum heat drying (VHD), and vacuum freeze drying (VFD). The quality assessment was based on unsaturated fatty acid (UFA) content, iodine value, estradiol, vitamin E, saponification value, thiobarbituric acid value (TBA), acid value, and antioxidant properties. The results showed that VFD had higher UFA content and lower acid value (139.950 ± 0.397 mg · g−1) and TBA value (0.097 ± 0.003) than others. VFD and NAD had higher iodine values (147.166 ± 1.475 and 146.803 ± 1.209, respectively) than others. There were no significant differences in estradiol, vitamin E, peroxide value and the clearance rates of ABTS+ free radicals and DPPH• free radicals with different drying methods. The findings will provide valuable insights into the industrialization of RCO.Graphical

Antimicrobial peptide 2K4L inhibits the inflammatory response in macrophages and Caenorhabditis elegans and protects against LPS-induced septic shock in mice

2K4L is a rationally designed analog of the short α-helical peptide temporin-1CEc, a natural peptide isolated and purified from the skin secretions of the Chinese brown frog Rana chensinensis by substituting amino acid residues. 2K4L displayed improved and broad-spectrum antibacterial activity than temporin-1CEc in vitro. Here, the antibacterial and anti-inflammatory activities of 2K4L in macrophages, C. elegans and mice were investigated. The results demonstrated that 2K4L could enter THP-1 cells to kill a multidrug-resistant Acinetobacter baumannii strain (MRAB 0227) and a sensitive A. baumannii strain (AB 22933), as well as reduce proinflammatory responses induced by MRAB 0227 by inhibiting NF-κB signaling pathway. Similarly, 2K4L exhibited strong bactericidal activity against A. baumannii uptake into C. elegans, extending the lifespan and healthspan of the nematodes. Meanwhile, 2K4L alleviated the oxidative stress response by inhibiting the expression of core genes in the p38 MAPK/PMK-1 signaling pathway and downregulating the phosphorylation level of p38, thereby protecting the nematodes from damage by A. baumannii. Finally, in an LPS-induced septic model, 2K4L enhanced the survival of septic mice and decreased the production of proinflammatory cytokines by inhibiting the signaling protein expression of the MAPK and NF-κB signaling pathways and protecting LPS-induced septic mice from a lethal inflammatory response. In conclusion, 2K4L ameliorated LPS-induced inflammation both in vitro and in vivo.

Seasonal changes in vitamin A metabolism-related factors in the oviduct of Chinese brown frog (Rana dybowskii)

The oviduct of the Chinese brown frog (Rana dybowskii) expands during pre-brumation rather than the breeding period, exhibiting a special physiological feature. Vitamin A is essential for the proper growth and development of many organisms, including the reproductive system such as ovary and oviduct. Vitamin A is metabolized into retinoic acid, which is crucial for oviduct formation. This study examined the relationship between oviducal expansion and vitamin A metabolism. We observed a significant increase in the weight and diameter of the oviduct in Rana dybowskii during pre-brumation. Vitamin A and its active metabolite, retinoic acid, notably increased during pre-brumation. The mRNA levels of retinol binding protein 4 (rbp4) and its receptor stra6 gene, involved in vitamin A transport, were elevated during pre-brumation compared to the breeding period. In the vitamin A metabolic pathway, the mRNA expression level of retinoic acid synthase aldh1a2 decreased significantly during pre-brumation, while the mRNA levels of retinoic acid α receptor (rarα) and the retinoic acid catabolic enzyme cyp26a1 increased significantly during pre-brumation, but not during the breeding period. Immunohistochemical results showed that Rbp4, Stra6, Aldh1a2, Rarα, and Cyp26a1 were expressed in ampulla region of the oviduct. Western blot results indicated that Aldh1a2 expression was lower, while Rbp4, Stra6, RARα, and Cyp26a1 were higher during pre-brumation compared to the breeding period. Transcriptome analyses further identified differential genes in the oviduct and found enrichment of differential genes in the vitamin A metabolism pathway, providing evidences for our study. These results suggest that the vitamin A metabolic pathway is more active during pre-brumation compared to the breeding period, and retinoic acid may regulate pre-brumation oviductal expansion through Rarα-mediated autocrine/paracrine modulation.

Plasticity of metamorphic traits of tadpoles of Rana chensinensis (Anura: Ranidae): interactive effects of food level and water exchange

Plasticity of metamorphic traits of tadpoles of Rana chensinensis (Anura: Ranidae): interactive effects of food level and water exchange

Chensinin-1b Alleviates DSS-Induced Inflammatory Bowel Disease by Inducing Macrophage Switching from the M1 to the M2 Phenotype.

Inflammatory bowel disease (IBD) is a chronic relapsing inflammatory disorder with an increasing prevalence worldwide. Macrophage polarization is involved in the pathogenesis of IBD. Repolarization of macrophage has thus emerged as a novel therapeutic approach for managing IBD. Chensinin-1b, derived from the skin of Rana chensinensis, is a derivative of a native antimicrobial peptide (AMP). It shows anti-inflammatory effects in sepsis models and can potentially modulate macrophage polarization. The objective of this research was to study the role of chensinin-1b in macrophage polarization and dextran sulfate sodium (DSS)-induced colitis. RAW264.7 macrophages were polarized to the M1 phenotype using lipopolysaccharide (LPS) and simultaneously administered chensinin-1b at various concentrations. The ability of chenisnin-1b to reorient macrophage polarization was assessed by ELISA, qRT-PCR, and flow cytometry analysis. The addition of chensinin-1b significantly restrained the expression of M1-associated proinflammatory cytokines and surface markers, including TNF-α, IL-6, NO, and CD86, and exaggerated the expression of M2-associated anti-inflammatory cytokines and surface markers, including IL-10, TGF-β1, Arg-1, Fizz1, Chil3, and CD206. Mechanistically, via Western Blotting, we revealed that chensinin-1b induces macrophage polarization from the M1 to the M2 phenotype by inhibiting the phosphorylation of nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK). In mouse models of colitis, intraperitoneal administration of chensinin-1b alleviated symptoms induced by DSS, including weight loss, elevated disease activity index (DAI) scores, colon shortening, colonic tissue damage, and splenomegaly. Consistent with our in vitro data, chensinin-1b induced significant decreases in the expression of M1 phenotype biomarkers and increases in the expression of M2 phenotype biomarkers in the mouse colitis model. Furthermore, chensinin-1b treatment repressesed NF-κB phosphorylation in vivo. Overall, our data showed that chensinin-1b attenuates IBD by repolarizing macrophages from the M1 to the M2 phenotype, suggesting its potential as a therapeutic candidate for IBD.

Investigation of seasonal changes in lipid synthesis and metabolism-related genes in the oviduct of Chinese brown frog (Rana dybowskii).

A peculiar physiological characteristic of the Chinese brown frog (Rana dybowskii) is that its oviduct dilates during pre-brumation rather than during the breeding season. This research aimed to examine the expression of genes connected with lipid synthesis and metabolism in the oviduct of R. dybowskii during both the breeding season and pre-brumation. We observed significant changes in the weight and size of the oviduct between the breeding season and pre-brumation. Furthermore, compared to the breeding season, pre-brumation exhibited significantly lower triglyceride content and a marked increase in free fatty acid content. Immunohistochemical results revealed the spatial distribution of triglyceride synthase (Dgat1), triglyceride hydrolase (Lpl and Hsl), fatty acid synthase (Fasn), and fatty acid oxidases (Cpt1a, Acadl, and Hadh) in oviductal glandular cells and epithelial cells during both the breeding season and pre-brumation. While the mRNA levels of triglycerides and free fatty acid synthesis genes (dgat1 and fasn) did not show a significant difference between the breeding season and pre-brumation, the mRNA levels of genes involved in triglycerides and free fatty acid metabolism (lpl, cpt1a, acadl, acox and hadh) were considerably higher during pre-brumation. Furthermore, the R. dybowskii oviduct's transcriptomic and metabolomic data confirmed differential expression of genes and metabolites enriched in lipid metabolism signaling pathways during both the breeding season and pre-brumation. Overall, these results suggest that alterations in lipid synthesis and metabolism during pre-brumation may potentially influence the expanding size of the oviduct, contributing to the successful overwintering of R. dybowskii.

Effects of perchlorate and exogenous T4 exposures on body condition and endochondral ossification of Rana chensinensis tadpoles

Perchlorate, as an endocrine-disrupting chemical (EDC), is largely produced and used in the military, fireworks, fertilizers, and other industries and widely exists in water. Although perchlorate is known to destroy the normal function of thyroid hormones (THs) in amphibians and interfere with their growth and development, the impact of TH levels caused by sodium perchlorate (NaClO4) on endochondral ossification and skeletal development is poorly investigated, and the underlying molecular mechanism has not been clarified. The present study aimed to explore the potential effects of NaClO4 and exogenous thyroxine (T4) on the skeletal development of Rana chensinensis tadpoles and elucidate the related molecular mechanisms. Our results showed that histological changes occurred to the femur and tibia-fibula of tadpoles raised in 250 μg/L NaClO4 and 5 μg/L exogenous T4, and the length of their hindlimbs was significantly reduced. In addition, exogenous T4 exposure significantly interfered with the expression of Dio3, TRβ, MMP9, MMP13, and Runx2, inhibiting the endochondral ossification process. Therefore, we provide robust evidence that the changes in TH levels caused by NaClO4 and exogenous T4 will adversely affect the endochondral ossification and skeletal development of R. chensinensis tadpoles.

Antimicrobial peptide 2K4L disrupts the membrane of multidrug-resistant Acinetobacter baumannii and protects mice against sepsis.

Antimicrobial peptides represent a promising therapeutic alternative for the treatment of antibiotic-resistant bacterial infections. 2K4L is a rationally-designed analog of a short peptide temporin-1CEc, a natural peptide isolated and purified from the skin secretions of the Chinese brown frog Rana chensinensis by substituting amino acid residues. 2K4L adopt an α-helical confirm in a membrane-mimetic environment and displayed an improved and broad-spectrum antibacterial activity against sensitive and multidrug-resistant Gram-negative and Gram-positive bacterial strains. Here, the action mechanism of 2K4L on multidrug resistant Acinetobacter baumannii (MRAB) and protection on MRAB-infected mice was investigated. The results demonstrated high bactericidal activity of 2K4L against both a multidrug resistant A. baumannii 0227 strain (MRAB 0227) and a sensitive A. baumannii strain (AB 22934), indicating a potential therapeutic advantage of this peptide. Strong positively-charged residues significantly promoted the electrostatic interaction on 2K4L with lipopolysaccharides (LPS) of the bacterial outer membrane. High hydrophobicity and an α-helical confirm endowed 2K4L remarkably increase the permeability of A. baumannii cytoplasmic membrane by depolarization of membrane potential and disruption of membrane integration, as well as leakage of fluorescein from the liposomes. Additionally, 2K4L at low concentrations inhibited biofilm formation and degraded mature 1-day-old MRAB 0227 biofilms by reducing the expression of biofilm-related genes. In an invasive A. baumannii infection model, 2K4L enhanced the survival of sepsis mice and decreased the production of the proinflammatory cytokines downregulating the phosphorylation level of signaling protein in MAPK and NF-κB signaling pathways, indicating that 2K4L represents a novel therapeutic antibiotic candidate against invasive multidrug-resistant bacterial strain infections.

The expression of cholesterol synthesis and steroidogenic markers in females of the Chinese brown frog (Rana dybowskii) during pre-spawning and pre-hibernation.

The oviduct of the Chinese brown frog (Rana dybowskii) expands in pre-hibernation rather than in pre-spawning, which is one of the physiological phenomena that occur in preparation for hibernation. Steroid hormones are known to regulate oviductal development. Cholesterol synthesis and steroidogenesis may play important role in the expansion of the oviduct before hibernation. In this study, we investigated the expression patterns of the markers that are involved in the de novo steroid synthesis pathway in the oviduct of R. dybowskii during pre-spawning and pre-hibernation. According to histological analysis, the oviduct of R. dybowskii contains epithelial cells, glandular cells, and tubule lumens. During pre-hibernation, oviductal pipe diameter and weight were significantly larger than during pre-spawning. 3-hydroxy-3-methylglutaryl CoA reductase (HMGCR), Low-density lipoprotein receptor (LDLR), steroidogenic acute regulatory protein (StAR), cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc), and steroidogenic factor 1 (SF-1) were detected in epithelial cells in pre-hibernation and glandular cells during pre-spawning. HMGCR, LDLR, StAR, and P450scc protein expression levels were higher in pre-hibernation than during pre-spawning, but the SF-1 protein expression level did not significantly differ. HMGCR, LDLR, StAR, P450scc (CYP11A1), and SF-1 (NR5A1) mRNA expression levels were significantly higher in pre-hibernation compared to pre-spawning. The transcriptome results showed that the steroid synthesis pathway was highly expressed during pre-hibernation. Existing results indicate that the oviduct is able to synthesize steroid hormones using cholesterol, and that steroid hormones may affect the oviductal functions of R. dybowskii.

The plasticity of metamorphic traits in the Chinese brown frog tadpoles fails to obey Richards' hypothesis

According to Richards' hypothesis, algae or cells in the intestinal tract has been considered the cellular inhibitory factor, when they fall off and hide in the faeces, excreted together with the faeces. If the animals feed on these faeces with algae or cells, and bring them into their systems, then the cellular inhibitory factor would play the main responsibility for growth inhibition. Here, we surveyed the effects of different combinations of faeces and food level on growth rates, survivorship, larval age and mass, and SVL at metamorphosis of the Chinese brown frog Rana chensinensis. Our results showed that food level can influence the length of the larval period of Chinese brown frog tadpoles, suggesting that delayed metamorphosis is caused by low food supply, indicative of a function of effective energy. Our data also clearly indicated that tadpoles in the presence of faeces were on average larger in body mass than those in the absence of faeces, which failed to obey Richards' hypothesis. Moreover, our results found evidence that faeces have a positive effect on the growth rate of tadpoles. Thus, there is no evidence for Richards' hypothesis, suggesting that this novel mechanism is selected for where coprophagy is likely to prove profitable, irrespective of the abundance of alternative food. Keywords: Rana chensinensis, Richards' hypothesis, faecal material, mass at metamorphosis, growth rate

Impact of breeding environments on the fatty acid composition of Rana chensinensis ovum from Changbai Mountain based on UPLC-Q/TOF-MS analysis

Rana chensinensis ovum (RCO), similar to fish roe, is a part of the gonads of Rana chensinensis and has become a premium delicacy due to its unique flavor and high nutritional value. The impact of breeding environments on the fatty acid content of RCO from various locations, primarily produced in Changbai Mountain, was explored in this study. RCO samples were collected from nine distinct regions that covered the primary production areas. Environmental factors such as altitude, longitude, latitude, average annual temperature, average annual ground temperature, annual precipitation, sunshine hours, soil top/substrate pH (T/S-pH-H2O), and soil top/substrate organic carbon content (T/S-OC) were evaluated. A UPLC-Q/TOF-MS-based method was used to determine the composition and distribution of fatty acids in RCO samples collected from different locations. Twenty-three fatty acid components, including DHA, EPA, DPA, ALA, C20:2, C20:3, C18:4, LA, C20:1, AA, C7:0, C10:1, C11:1, C16:2, C16:1, C16:0, OA, C18:0 C19:0, C23:1, C24:1, C22:0, and C17:1, were identified in RCO samples. Gray relational analysis (GRA) and Pearson correlation were employed to evaluate the influence of each environmental factor on fatty acid composition. Hierarchical cluster analysis (HCA), principal component analyses (PCA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to classify RCO samples from different origins and analyze their similarities and differences. The results indicated that the habitat of Rana chensinensis can influence their nutritional value, as differences in fatty acid components were observed in RCO samples collected from different environments.

Effects of perchlorate and exogenous T4 on growth, development and tail resorption of Rana chensinensis

Endocrine disruptors have been demonstrated to exert adverse effects on growth and development of amphibians by disrupting hormone levels. Tail resorption, which is one of the most remarkable events during amphibian metamorphosis, is closely associated with thyroid hormones levels. However, limited research has been conducted on the effects of endocrine disruptors on tail resorption in amphibians. This study explored the effects of NaClO4 and T4 on the growth, development and tail resorption during the metamorphosis of Rana Chensinensis. The results demonstrated that exposure to NaClO4 led to an increase in body size and a delay in metamorphosis of R. Chensinensis tadpoles. Histological analysis revealed that both NaClO4 and exogenous T4 exposure resulted in thyroid gland injury, and NaClO4 treatment delayed the degradation of notochord and muscles, thereby delaying tail resorption. Moreover, transcriptome sequencing results showed that apoptosis-related genes (APAF1, BAX and CASP6) and cell component degradation-related genes (MMP9 and MMP13) were highly expressed in the T4 exposure group, and the expression of oxidative stress-related genes (SOD and CAT) was higher in the NaClO4 exposure group. Taken together, both NaClO4 and exogenous T4 affect tail resorption in R. Chensinensis, thereby affecting their adaptation to terrestrial life. The present study will not only provide a reference for future experimental research on the effects of other endocrine disruptors on the growth, development and tail resorption of amphibians but will also provide insights into environmental protection and ecological risk assessment.

The antimicrobial peptide chensinin-1b alleviates the inflammatory response by targeting the TLR4/NF-κB signaling pathway and inhibits Pseudomonas aeruginosa infection and LPS-mediated sepsis

Excessive inflammatory responses are a major contributor to the high mortality associated with sepsis, a prevalent global complication. Therefore, the potential therapeutic strategy for sepsis involves targeting macrophages and reducing proinflammatory cytokine release. Chensinin-1b, an analog of the natural antimicrobial peptide derived from Rana chensinensis skin secretion, exhibits broad-spectrum antibacterial activity and adopts a random coil conformation in both PBS and membrane solution. By efficiently neutralizing LPS, chensinin-1b holds promise in alleviating LPS-induced inflammatory responses. In this study, we established a mouse septic shock model by exposing mice to multiple-drug-resistant Pseudomonas aeruginosa, as well as an endotoxin-mediated sepsis model induced by LPS. Administering chensinin-1b significantly prolonged the survival of the experimental mice, concurrently mitigating inflammatory responses and reducing organ damage. Additionally, we investigated the anti-inflammatory mechanism of chensinin-1b using a constructed LPS-induced mouse macrophage RAW264.7 inflammatory model. Our findings demonstrated that chensinin-1b effectively mitigated the excessive activation of the TLR4/NF-κB signaling pathway by directly neutralizing extracellular LPS, thus ameliorating the inflammatory response. Moreover, upon blocking the TLR4 signaling pathway, chensinin-1b further reduced the release of proinflammatory cytokines induced by LPS, indicating alternative modes of regulation. Notably, chensinin-1b rapidly entered RAW264.7 cells within 30 min via endocytosis, diffusing into the cytoplasm while retaining its anti-inflammatory properties intracellularly. Although further investigations are warranted to comprehensively elucidate the intracellular anti-inflammatory mechanism of chensinin-1b, our findings substantiate its possession of anti-inflammatory properties both intracellularly and extracellularly. Thus, chensinin-1b emerges as a promising candidate for mitigating excessive inflammatory responses associated with sepsis.

Regulation mechanisms underlying tail resorption in Bufo gargarizans metamorphosis

Anurans have been excellent organisms for studying amphibian metamorphosis. Tail resorption is a remarkable event that occurs during amphibian metamorphosis. Although tail resorption has been previously studied in other anurans like Xenopus laevis and Rana chensinensis, there is no report on Bufo gargarizans. This paper thus explored the mechanism of tail resorption during metamorphosis in Bufo gargarizans tadpoles through some biological research methods. Histological results showed that the tail tissues of tadpoles gradually degraded as metamorphosis progressed. RNA sequencing analysis was performed to examine the expression level and functional enrichment of differentially expressed genes in the tail. In addition, we analyzed the mRNA expression levels of genes related to tail resorption by quantitative real-time polymerase chain reaction. We also speculated on three pathways that participate in the regulation of tail resorption based on the above results. The present study might provide a theoretical basis and novel insights for further research of complex molecular mechanisms of tail resorption in amphibians.

Effects of cadmium and diethylhexyl phthalate on skin microbiota of Rana chinensis tadpoles.

Skin microbiotas play a crucial role in the health, homeostasis, and immune function of amphibians. The contaminants in water could affect the structure and composition of microbial communities. The effects of coexisting pollutants on frogs cannot be adequately explained by a single exposure due to the coexistence of Cd and DEHP in the environment. Following exposure to Cd and/or DEHP, we examined the histological characteristics of Rana chensinensis tadpoles. We also used the 16S rRNA gene sequencing technique to assess the relative abundance of skin microbial communities among tadpoles from each treatment group. Our findings indicate that R. chensinensis' skin experienced some degree of injury due to exposure to Cd and DEHP, which led to the imbalance of their skin microbial community homeostasis and thus interfered with the normal trial status of the host. That may eventually lead to the decline of the amphibian population.

Intestinal response of Rana chensinensis larvae exposed to Cr and Pb, alone and in combination

Although numerous investigations on the adverse impact of Cr and Pb have been performed, studies on intestinal homeostasis in amphibians are limited. Here, single and combined effects of Cr (104 μg/L) and Pb (50 μg/L) on morphological and histological features, bacterial community, digestive enzymes activities, as well as transcriptomic profile of intestines in Rana chensinensis tadpoles were assessed. Significant decrease in the relative intestine length (intestine length/snout-to-vent length, IL/SVL) was observed after exposure to Pb and Cr/Pb mixture. Intestinal histology and digestive enzymes activities were altered in metal treatment groups. In addition, treatment groups showed significantly increased bacterial richness and diversity. Tadpoles in treatment groups were observed to have differential gut bacterial composition from controls, especially for the abundance of phylum Proteobacteria, Firmicutes, Verrucomicrobia, Actinobacteria, and Fusobacteria as well as genus Citrobacter, Anaerotruncus, Akkermansia, and Alpinimonas. Moreover, transcriptomic analysis showed that the transcript expression profiles of GPx and SOD isoforms responded differently to Cr and/or Pb exposure. Besides, transcriptional activation of pro-apoptotic and glycolysis-related genes, such as Bax, Apaf 1, Caspase 3, PK, PGK, TPI, and GPI were detected in all treatment groups but downregulation of Bcl2 in Pb and Cr/Pb mixture groups. Collectively, these results suggested that Cr and Pb exposure at environmental relevant concentration, alone and in combination, could disrupt intestinal homeostasis of R. chensinensis tadpoles.

Effects of temperature on metamorphosis and endochondral ossification in Rana chensinensis tadpoles

Temperature is one of the important factors affecting the growth, development, and metamorphosis of amphibians. Endochondral ossification during metamorphosis plays a crucial role in amphibian survival and adaptation on land. In this study, we explored the effects of different temperature treatments on the growth, development, and endochondral ossification of Rana chensinensis tadpoles during metamorphosis. The results showed that high temperature exposure may affect the skeletal development of tadpoles during metamorphosis, such as reduction of bone length and ossification of limbs, thyroid gland damage and change of ossification-related genes expression levels，and ultimately affect the movement and survival of tadpoles in the terrestrial environment. These results provide an experimental reference for further research on the effects of temperature on amphibian growth and development and provide an important theoretical basis for the decline of the amphibian population caused by temperature.

Facile construction of electrospun zein nanofiber loaded with Rana chensinensis skin collagen for wound care after caesarean section surgery

Numerous types of biomedical applications have found success using electrospun nanofibers. However, electrospinning techniques overlook Zein fibers favoring monolithic and coaxial fibers. Uniaxial electrospinning was used to construct Zein-structured nanofibers for wound healing, with the phase separation of electrospun polycaprolactone (PCL)/polyvinyl alcohol (PVA) in solution, providing the basis for the investigation. Successful loading of silver nanoparticles (AgNPs) and Rana chensinensis skin collagen (RCSCs) into Zein nanofibers (NFs) improved their bioactivity and antibacterial activity as wound dressings. Zein nanofibers were investigated for their heat conductivity, wettability, and mechanical characteristics. The cytotoxic effects of fabricated nanofibers were examined by using L929 fibroblast cells. Ag@RCSCs-NFs also increased cell migration and proliferation, and the wound scratch model was significantly reduced in size using an in vitro scratch assay. The antibacterial studies showed that the Ag@RCSCs-NFs have a potent antibacterial effect against the tested microbial pathogens (Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus)). Finally, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of PCL/PVP, RCSCs, and Ag@RCSCs against two different bacteria were determined. These results suggest that wound care using Zein nanofibers loaded with RCSCs and AgNPs during cesarean section surgery has great promise.

Effects of temperature on intestinal microbiota and lipid metabolism in Rana chensinensis tadpoles.

Climate change such as global warming is considered a major threat to amphibians. The guts of amphibians are home to trillions of microbes, which are key regulators of gastrointestinal digestion and play a crucial role in lipid metabolites. The aim of this study was to evaluate the effect of temperature change on intestinal microbiota and lipid metabolism in Rana chensinensis tadpoles. Morphological and intestinal microbiota data of R. chensinensis larvae exposed to different temperatures (15°C, 21°C, and 26°C) were measured. The results show that the warm temperature causes histological damage to the intestinal epithelium. In addition, temperature treatments alter the diversity and composition of gut microbes in R. chensinensis tadpoles. At the phylum level of intestinal microbial community, Campilobacterota was detected only in the warm group. At the genera level, unclassified_f__Enterobacteriaceae was markedly declined in the warm group but was notably enriched in the cold group. For lipid metabolism-related genes, the expression levels of GPR109A, HDAC1, and APOA-I decreased significantly in both warm and cold treatment groups, while the expression levels of CLPS and LIPASE increased significantly. Collectively, these observations demonstrated that warm and cold temperatures may reduce the immune capacity of tadpoles by changing the composition of intestinal microorganisms and the expression of genes related to lipid metabolism, affecting the survival of tadpoles.