Transcriptome analysis of Fenneropenaeus merguiensis in response to Vibrio proteolyticus infection.

In recent years, due to the destruction of the culture environment and serious ecological pressure, especially in the process of culture, residual bait, faeces and fishery drug abuse will lead to the accumulation of harmful metabolites such as ammonia nitrogen and nitrite, and biological denitrification is the most economical and effective method to remove the single. Therefore, in this study, a nitrite removal strain XA19 was isolated and screened from a shrimp biofloc culture pond. This strain was identified as a clade of Vibrio proteolyticus because the homology between XA19 and V. proteolyticus WDVP was as high as 99.86% by using 16S rDNA gene sequence analysis and NCBI database comparison. Scanning electron microscopy images showed that V. proteolyticus is short-rod-shaped with a curved body and no budding spores, pods and flagella. Antimicrobial susceptibility test proved that V. proteolyticus was resistant to ampicillin, oxacillin, penicillin, vancomycin and clindamycin. In the median lethal concentration 50 (LC50 ) test, at 7-day post-infection (dpi), LC50 of V. proteolyticus for Fenneropenaeus merguiensis was 1.69 × 104 CFU/mL. Transcriptome sequencing analysis was carried out on hepatopancreas of F. merguiensis at 24 and 48 hpi. A total of 176 differentially expressed genes (DEGs) were screened at 24 hpi, including 104 up-regulated DEGs and 72 down-regulated DEGs, and a total of 52 DEGs were screened at 48 hpi, including 32 up-regulated DEGs and 20 down-regulated DEGs. In the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of DEGs, many immune-related signalling pathways were significantly enriched, including Hippo signalling pathway, phagosome, Toll and Imd signalling pathways and Wnt signalling pathway. In addition, some pathways related to Warburg effect were also enriched, including Glycolysis/Gluconeogenesis, Biosynthesis of amino acids, amino sugar and nucleotide sugar metabolism and so on. In this study, the toxicity and drug sensitivity of V. proteolyticus were systematically studied, and the immune response of hepatopancreas of F. merguiensis to V. proteolyticus infection was preliminarily revealed from the molecular level. The results may provide a reference for the prevention and control of V. proteolyticus.

Integrated analysis of intestinal microbiota and metabolomic reveals that decapod iridescent virus 1 (DIV1) infection induces secondary bacterial infection and metabolic reprogramming in Marsupenaeus japonicus.

In recent years, with global warming and increasing marine pollution, some novel marine viruses have become widespread in the aquaculture industry, causing huge losses to the aquaculture industry. Decapod iridescent virus 1 (DIV1) is one of the newly discovered marine viruses that has been reported to be detected in a variety of farmed crustacean and wild populations. Several previous studies have found that DIV1 can induce Warburg effect-related gene expression. In this study, the effects of DIV1 infection on intestinal health of shrimp were further explored from the aspects of histological, enzymatic activities, microorganisms and metabolites using Marsupenaeus japonicus as the object of study. The results showed that obvious injury in the intestinal mucosa was observed after DIV1 infection, the oxidative and antioxidant capacity of the shrimp intestine was unbalanced, the activity of lysozyme was decreased, and the activities of digestive enzymes were disordered, and secondary bacterial infection was caused. Furthermore, the increased abundance of harmful bacteria, such as Photobacterium and Vibrio, may synergized with DIV1 to promote the Warburg effect and induce metabolic reprogramming, thereby providing material and energy for DIV1 replication. This study is the first to report the changes of intestinal microbiota and metabolites of M. japonicus under DIV1 infection, demonstrating that DIV1 can induce secondary bacterial infection and metabolic reprogramming. Several bacteria and metabolites highly associated with DIV1 infection were screened, which may be leveraged for diagnosis of pathogenic infections or incorporated as exogenous metabolites to enhance immune response.

Integrated Analysis of mRNA-Seq and MiRNA-Seq Reveals the Molecular Mechanism of the Intestinal Immune Response in Marsupenaeus japonicus Under Decapod Iridescent Virus 1 Infection.

The intestine is not only an important digestive organ but also an important immune organ for shrimp; it plays a key role in maintaining homeostasis. Decapod iridescent virus 1 (DIV1) is a new type of shrimp-lethal virus that has received extensive attention in recent years. To date, most studies of the shrimp intestinal immune response under viral infections have relied on single omics analyses; there is a lack of systematic multi-omics research. In the current study, intestinal mRNA-seq and microRNA (miRNA)-seq analyses of Marsupenaeus japonicus under DIV1 infection were performed. A total of 1,976 differentially expressed genes (DEGs) and 32 differentially expressed miRNAs (DEMs) were identified. Among them, 21 DEMs were negatively correlated with 194 DEGs from a total of 223 correlations. Functional annotation analysis revealed that M. japonicus can regulate glycosaminoglycan biosynthesis (chondroitin sulfate, dermatan sulfate, and keratan sulfate), vitamin metabolism (retinol metabolism and ascorbate and aldarate metabolism), immune pathway activation (Toll and IMD signaling pathways, Wnt signaling pathway, IL-17 signaling pathway, and Hippo signaling pathway), immunity enzyme activity promotion (triose-phosphate isomerase), antimicrobial peptide (AMP) expression, reactive oxygen species (ROS) production, and cell apoptosis through miRNAs to participate in the host's antiviral immune response, while DIV1 can influence Warburg effect-related pathways (pyruvate metabolism, glycolysis/gluconeogenesis, and citrate cycle), glycosphingolipid biosynthesis-related pathways (glycosphingolipid biosynthesis-globo and isoglobo series and glycosphingolipid biosynthesis-lacto and neolacto series), and the tight junction and adhesion junction of the intestinal mucosal epithelium through the host's miRNAs and mRNA to promote its own invasion and replication. These results indicate that intestinal miRNAs play important roles in the shrimp immune response against DIV1 infection. This study provides a basis for further study of the shrimp intestinal antiviral immune response and for the formulation of effective new strategies for the prevention and treatment of DIV1 infection.

Research into the hemocyte immune response of Fenneropenaeus merguiensis under decapod iridescent virus 1 (DIV1) challenge using transcriptome analysis.

The banana shrimp (Fenneropenaeus merguiensis) is a common cultural species worldwide. With the development of the shrimp farming industry, increasing number of diseases have emerged and cause huge impacts. Decapod iridescent virus 1 (DIV1) is a new virus of the family Iridoviridae isolated in China that causes very high mortality in shrimp. In this study, DIV1 and PBS were injected into two groups of shrimp, and hemocytes were collected for comparative transcriptomic analysis. We confirmed that F. merguiensis was the new host of DIV1 by nested PCR. A total of 100,759 unigenes were assembled from the control group and the DIV1 infected group, with an average length of 733.06 bp and N50 of 1136 bp. Significant hits were found in 21,465 unigenes compared to known sequences in major databases including COG (33.30%), GO (42.17%), KEGG (46.76%), KOG (61.37%), Pfam (66.90%), Swissprot (54.21%) and Nr (93.86%). A total of 1003 differentially expressed genes (DEGs) were identified, including 929 up-regulated genes and 74 down-regulated genes. Several known immune-related genes, including caspase, C-type lectin, Wnt5 and integrin, were among the differentially expressed transcripts. A total of 14,459 simple sequence repeats, including 8128 monomers, 3276 dimers, 1693 trimers, 150 quadmers, 4 pentamers and 16 hexamers, were found in the transcriptomic dataset. Our study is the first comprehensive investigation of the transcriptomic response to DIV1 infection in F. merguiensis. Collectively, these results not only provide valuable information for characterizing the immune mechanisms of the shrimp responses to DIV1 infection, they open new ways for the study of the molecular mechanisms of DIV1 infection in F. merguiensis.

Acute Beneficial Effects of Sodium Nitroprusside in a Rabbit Model of Massive Pulmonary Embolism Associated with Circulatory Shock.

We established a rabbit model of acute massive pulmonary embolism (PE) with associated circulatory shock using autologous blood clots. Rabbits were randomly assigned to a sham operation group (S group), model group (M group; equal volume of saline intravenously after shock), and sodium nitroprusside group (SNP group; sodium nitroprusside intravenously after shock). SNP treatment significantly decreased mean pulmonary arterial pressure and increased mean arterial pressure and arterial partial pressure of oxygen and resulted in a partial reversal of the acute circulatory failure. The shock-reversal rate was 0% in the M group and 80% in the SNP group. Moreover, pulmonary artery angiography and echocardiography examinations evidenced alleviated PE-induced changes after SNP therapy. 5-Hydroxytryptamine was significantly reduced in both PE and non-PE tissues, thromboxane A2 level was significantly reduced in PE and tended to be lower in non-PE tissues, neutrophil accumulation was significantly reduced in both PE and non-PE tissues after SNP therapy. Our study demonstrated that pulmonary vasospasm in the nonembolic region might be a major pathologic factor leading to reduced left ventricular filling and circulatory shock after massive PE. Reduction of pulmonary vasospasm in the nonembolic area after SNP might serve as a major therapeutic mechanism involved in the observed beneficial effects of SNP in this model.

Mandible reconstruction assisted by preoperative virtual surgical simulation.

OBJECTIVE: In this study, we evaluated the clinical efficacy of mandible reconstruction with preoperative virtual planning, which focused on esthetics and occlusion. STUDY DESIGN: A series of 9 patients were enrolled prospectively to undergo mandibulectomy and simultaneous reconstruction. Preoperative spiral CT scans of the maxillofacial region and the fibula region were performed. Virtual surgery of tumor resection and fibula reconstruction was performed in the Mimics platform. The reconstructed mandible models were fabricated with CAD/CAM technique. The reconstruction plate and the positioning template were accommodated to the stereolithographic model as the surgical template. RESULTS: Surgery was performed accurately according to the templates. All the fibula flaps survived. The appearance and occlusion of the patients were satisfactory. CONCLUSIONS: With preoperative virtual planning, the spatial relationship of the mandible and the fibula graft can be planned individually, which helps achieve optimum appearance and occlusion relationship.

CT virtual sialendoscopy versus conventional sialendoscopy in the visualization of salivary ductal lumen: an in vitro study.

OBJECTIVES/HYPOTHESIS: To evaluate the feasibility of computed tomographic (CT) virtual sialendoscopy and its performance characteristics for the visualization of salivary ductal lumen as compared with conventional sialendoscopy. STUDY DESIGN: An in vitro study. METHODS: Conventional sialendoscopy was performed in 16 submandibular glands in vitro and the surgical findings were recorded. Then a contrast agent was injected into the Wharton's duct via orifice. After CT scanning, the CT data were transferred to an independent workstation and were postprocessed with commercially available software to generate three-dimensional reconstructive and virtual sialendoscopic images. Finally, the plastination of submandibular glands was performed with 20% perchloroethylene ethyl acetate plastics filler to make cast specimens. CT three-dimensional reconstruction of the ductal system was compared with the cast specimen, and the performance of conventional sialendoscopy and CT virtual sialendoscopy for the visualization of salivary ductal lumen was also compared. RESULTS: The CT data acquisition and postprocessing protocol were feasible. CT three-dimensional reconstruction enabled clear visualization of the ductal system from any directions and angles, which showed close resemblance to the cast specimens. And CT virtual sialendoscopy created clear endoluminal views of salivary ducts simulating those obtained with conventional sialendoscopy. CONCLUSIONS: Our initial experience shows that CT virtual sialendoscopy has the potential to be an attractive imaging technique for the visualization of salivary ductal lumen.