Chitosan and its derivatives such as low molecular weight chitosans (LMWCs) have been reported to exert many biological activities, such as antioxidant and antitumor effects. However, complex and molecular weight dependent effects of chitosan remain controversial and the mechanisms that mediate these complex effects are still poorly defined. This study was carried out to investigate the immunostimulative effect of different molecular weight chitosan in RAW264.7 macrophages. Our data suggested that two LMWCs (molecular weight of 3 kDa and 50 kDa) both possessed immunostimulative activity, which was dependent on dose and, at the higher doses, also on the molecular weight. LMWCs could significantly enhance the the pinocytic activity, and induce the production of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), interferon-γ (IFN-γ), nitric oxide (NO) and inducible nitric oxide synthase (iNOS) in a molecular weight and concentration-dependent manner. LMWCs were further showed to promote the expression of the genes including iNOS, TNF-α. Taken together, our findings suggested that LMWCs elicited significantly immunomodulatory response through up-regulating mRNA expression of proinflammatory cytokines and activated RAW264.7 macrophage in a molecular weight- and concentration-dependent manner.

The experiment was conducted in a sealed, temperature-controlled room. The initial water temperature of the treatment group was 12°C, lowered by 1°C a day to 8°C using a chiller, and maintained at that temperature for 3 days. The water temperature was then increased by 1°C per day to 12°C using a heater. The control group were the fish that kept at 12°C at the beginning of the experiment; T1 represents the group that sampled on the first day when the water temperature reached 8 from 12°C; T2 represents the group that sampled when the fish had been kept at 8°C for 3 days; T3 represents the group sampled when the water temperature was increased to 12 from 8°C. The antioxidant abilities of the liver, muscle, and intestine of large yellow croaker at different temperatures and times were determined, and the expression of HSP70 and HSP90 in the liver as well as the changes in intestinal microbiota were investigated. The results showed that the activities of SOD, CAT, GSH-Px, and MDA content increased significantly when the water temperature decreased from 12 to 8°C, significantly higher than those in the control group. The gene expression levels of HSP70 and HSP90 in the liver of large yellow croaker significantly increased under low temperature stress and were significantly higher than those in the control group. The diversity and abundance of intestinal microbiota of large yellow croaker were altered when the water temperature was maintained at 8°C, although the change was not obvious. Proteobacteria, Firmicutes, and Bacteroidetes were the dominate phyla in both the 12 and 8°C groups, and the temperature variation did not change the functional pathways of the microbiota. In conclusion, the acute temperature variation affected the antioxidant ability and heat shock protein expression of large yellow croaker but did not affect the diversity or abundance of the intestinal microbiota. These results will provide a theoretical basis for studying the mechanisms of large yellow croaker overwintering and adaptation to low temperatures.

The mitochondrial genome plays a crucial role in the balance of energy and heat production in organisms and, thus, may be under natural selection due to its potential role in adaptive divergence and speciation. Here, we studied natural selection on the mitogenome of Acrossocheilus (Cypriniformes: Cyprinidae), a genus of fish that inhabits a broad latitudinal distribution ranging from the tropics and subtropics through temperate regions. Specifically, we used 25 published mitogenome sequences of Acrossocheilus species to investigate phylogenetic relationships in this genus and detected signals of positive selection on 13 protein-coding, mitochondrial genes. We found that relaxed purifying selection and genetic drift were the predominant evolutionary forces acting on the analyzed mitogenomes. However, we also found evidence of diversifying selection on some codons, indicating episodes of positive selection. Additionally, we analyzed the mitogenomic data within an environmental modeling framework and found that the Ka/Ks ratio of ATP6 may correlated with a mean diurnal temperature range (p = 0.0449), while the Ka/Ks ratio of COX2 may correlated with precipitation during the driest month (p = 0.00761). These results suggest that the mitogenomes of Acrossocheilus species may be involved in evolutionary adaptations to different habitats. Based on this, we believe that our study provides a new insight into the role of the mitochondrial genome of Acrossocheilus species in adaptation to different environments. During our study, we also discovered several cases of paraphyly and polyphyly among accessions of species and their putative synonyms. Thus, our study suggests that a careful reassessment of the taxonomy of Acrossocheilus is using high-quality molecular data merited.

PFOA, a newly emerging persistent organic pollutant, is widely present in various environmental media. Previous reports have proved that PFOA exposure can accumulate in the ovary and lead to reproductive toxicity in pregnant mice. However, the potential mechanism of PFOA exposure on fertility remains unclear. In this study, we explore how PFOA compromises fertility in the zebrafish. The data show that PFOA (100 mg/L for 15 days) exposure significantly impaired fertilization and hatching capability. Based on tissue sections, we found that PFOA exposure led to ovarian damage and a decrease in the percentage of mature oocytes. Moreover, through in vitro incubation, we determined that PFOA inhibits oocyte development. We also sequenced the transcriptome of the ovary of female zebrafish and a total of 284 overlapping DEGs were obtained. Functional enrichment analysis showed that 284 overlapping DEGs function mainly in complement and coagulation cascades signaling pathways. In addition, we identified genes that may be associated with immunity, such as LOC108191474 and ZGC:173837. We found that exposure to PFOA can cause an inflammatory response that can lead to ovarian damage and delayed oocyte development.