Isolation and characterization of 20 polymorphic microsatellites loci for Xenocypris davidi based on high-throughput sequencing.

Xenocypris davidi is one of the most economically important freshwater fish in China. However, few molecular markers have been reported for this species, impeding in-depth population genetic, dispersal, and gene flow studies. In the present study, a batch of novel polymorphic microsatellites from the genome of X. davidi were isolated and characterized using high-throughput sequencing. A total of 20 microsatellite markers were isolated. Analysis of 33 individuals revealed an average of 7.35 alleles per locus, ranging from 3 to 18. The observed and expected heterozygosities ranged from 0.3 to 1 and from 0.426 to 0.93, respectively. Only one tested locus significantly deviated from Hardy-Weinberg equilibrium. 18 microsatellite loci were highly polymorphic (PIC > 0.5). These newly isolated microsatellite markers would be useful to study the population genetics and stock management of X. davidi.

Radial P34HB Electrospun Fiber: A Scaffold for Bone Tissue Engineering.

Finding suitable scaffold material is always an enormous challenge in the study of bone tissue engineering. Designing and preparing bone scaffolds with biomimetic properties is also a difficult problem for bone reparation projects. This project intends to fabricate radial bio-plasticpoly-3-hydroxybutyrate-4-hydroxybutyrate (P34HB) electrospun fibers scaffold that mimic structural, compositional and stiffness properties via an electro spinning technique. The surface morphology, hydrophilicity of the radial fibers scaffold were tested, a contact angle meter and a universal material tester. Bone marrow mesenchymal stem cell (BMSC) morphologies on radial P34HB electrospun fibers scaffold were observed after cell culture under fluorescence microscopy. Tests of cell viability on radial P34HB electrospun fibers scaffold were conducted. We further tested the osteogenic differentiation ability of radial fibers scaffold. These results showed that radial P34HB electrospun fibers scaffold have good biosafety, biocompatibility and osteogenic induction. The radial structure of the scaffold also has a strong effect on the induction of bone formation. Moreover, the structure could also improve the bone contact area of the implant and increase the locking and fixation between the implant and the bone. We plan to apply this procedure to animal experiments for bone defect repair in further research.

Acute toxicity of cypermethrin on the juvenile of red claw crayfish Cherax quadricarinatus.

In order to assess the toxicity of Cypermethrin (CYP), the 50% lethal concentration (LC50) of CYP on the juvenile of Cherax quadricarinatus is assessed. Meanwhile, the transcription level and the content in the antioxidant and biotransformation enzymes in hepatopancreas and immune enzymes in the serum of C. quadricarinatus exposed to CYP (0.1, 1, 10 and 100 ng·L-1) for 96 h were analyzed to reveal the CYP toxicity and detoxification mechanism. 24, 48, 72, 96 h LC50 were 1305.14, 424.52, 287.10 and 215.99 ng·L-1, respectively. There was no significant change of the content of enzymes at low concentration (0.16 ng·L-1). The fast increase of SOD and CAT content was observed at early stage (24 h), subsequent decreased at later stage of trail at medium concentration (0.32 and 0.63 ng·L-1). However, high concentration (1.25 ng·L-1) of CYP significantly inhibited SOD and CAT content. There was a significant increase in the level of MDA, PC and the content of GPx, EROD, CarE, GST at medium and high concentration after 72 h and 96 h exposure. The Na+-K+-ATPase, PO, ALK content decreased at medium and high concentration, especially at the 72-h and the 96-h exposure. The transcription was altered similarly to enzyme content, but the transcriptional response was generally more immediate than enzymatic response. Heat shock protein (hsp70) and multidrug resistance-associated protein 2 (abcc2) genes were up-regulated.

De novo assembly and characterization of narrow-ridged finless porpoise renal transcriptome and identification of candidate genes involved in osmoregulation.

During the evolutionary transition from land to water, cetaceans have undergone numerous critical challenges, with osmoregulation being the major one. Two subspecies of the narrow-ridged finless porpoise (Neophocaena asiaeorientalis), the freshwater Yangtze finless porpoise (N. a. asiaeorientalis, NAA) and the marine East Asian finless porpoise (N. a. sunameri, NAS), provide excellent subjects to understand the genetic basis of osmoregulatory divergence between freshwater and marine mammals. The kidney plays an important and well-established role in osmoregulation in marine mammals and thus, herein, we utilized RNA-seq to characterize the renal transcriptome and preliminarily analyze the divergence between the NAA and the NAS. Approximately 48.98 million clean reads from NAS and 49.40 million clean reads from NAA were obtained by RNA-Seq. And 73,449 (NAS) and 68,073 (NAA) unigenes were assembled. Among these annotations, 22,231 (NAS) and 21,849 (NAA) unigenes were annotated against the NCBI nr protein database. The ion channel complex GO term and four pathways were detected as relevant to osmoregulation by GO and KEGG pathway classification of these annotated unigenes. Although the endangered status of the study species prevented analysis of biological replicates, we identified nine differentially expressed genes (DEGs) that may be vital in the osmoregulation of the narrow-ridged finless porpoise and worthwhile for future studies. Of these DEGs, the differential expression and distribution of the aquaporin-2 (AQP2) in the collecting duct were verified using immunohistochemical experiments. Together, this work is the first report of renal transcriptome sequencing in cetaceans, and it will provide a valuable resource for future molecular genetics studies on cetacean osmoregulation.