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The mucosa of vertebrates is a particularly complex but dynamic environment in which the host constantly interacts with trillions of commensal microorganisms and pathogens. Although the internal and external mucosal microbiomes with immune defense of mammals have been well investigated, the relationship between mucosal microbes and their host’s immune responses has not been systematically understood in the early vertebrates. In this study, we compared the composition and distribution of mucosal microbiota in common carp ( Cyprinus carpio ), and found that there were significant differences of microbiota between in the internal (gut) and external mucosal (buccal mucosa, gills and skin) tissues. Next, we successfully constructed an infection model with spring viremia of carp virus (SVCV). Specifically, following viral infection, the immune and antiviral related genes showed different up-regulation in all selected mucosal tissues while significant morphological changes were only found in external tissues including buccal mucosa, gills and skin. Using 16S rRNA gene sequence, we revealed that the abundance of Proteobacteria in mucosal tissues including buccal mucosa, gills and gut showed increased trend after viral infection, whereas the abundance of Fusobacteria significantly decreased in gut. In addition, the loss of dominant commensal microorganisms and increased colonization of opportunistic bacteria were discovered in the mucosal surfaces indicating that a secondary bacterial infection might occur in these mucosal tissues after viral infection. Overall, our results firstly point out the distribution of internal and external mucosal microbiota and analyze the changes of mucosal microbiota in common carp after SVCV infection, which may indicated that the potential role of mucosal microbiota in the antiviral process in early vertebrates.

The digestive tract is a unique series of organs that is inhabited by a range of commensal microbes while also exposed to an overwhelming load of dietary antigens. It is widely known that mammals have evolved complex and efficient immune strategies to protect the mucosa of the digestive tract. However, in the early vertebrates, the roles of mucosal immune defense and microbial communities in the different segments of the digestive tract are not well-understood. Here, we constructed a bath infection model with infectious hematopoietic necrosis virus (IHNV) in rainbow trout ( ). Importantly, following viral infection, we found that the IHNV distribution and the reactions of immune-related genes had similar trends that decreased across the digestive tract. Hematoxylin and eosin (H & E) and alcian blue (A & B) staining of the trout digestive tract showed that the pathological changes only occurred in the buccal and pharyngeal mucosal tissues. Moreover, the increased diversity of the microbial community was only detected in the buccal mucosa through 16S rRNA gene sequencing, suggesting that the magnitude of the immune response and microbial community changes are related to the IHNV load and the original microbial diversity. In addition, the loss of digestive tract dominant species and increased colonization of opportunistic bacteria were discovered in the buccal mucosal surface indicating that a secondary bacterial infection occurred in this mucosal tissue.

The gallbladder (GB) microbiota plays critical roles in mammalian metabolism and immune homeostasis, and its relationship with human disease has been extensively studied over the past decade. However, very little is known about the interplay between GB microbiota and the immune functions of teleost fish, the earliest bony vertebrate with a GB. Therefore, this study sought to investigate the composition of the teleost GB microbiota and the potential mechanisms through which it affects mucosal immunity. In our results, we found that the GB mucosa (GM) and bile bacterial community shared a similar microbiological composition with that of the gut mucosa in naïve individuals. IHNV infection induced a profound GB inflammation and disrupted their microbial homeostasis followed by a strong anti-bacterial response. Interestingly, beneficial bacteria from the Lactobacillales order showed a significant increase in the abundance of the bile microbial community, whereas the structure of the Mycoplasmatales order in the gut microbial community was markedly changed. All in all, our study characterized the structure of the GB microbial ecosystem in teleost fish, and the fish GB microbiome shared a high similarity with the gut microbiota. More importantly, our findings offer solid evidence that the teleost GB evolved immune functions to preserve its mucosal microbial homeostasis, suggesting that both the microbiota and mucosal immunity of the GB might have co-evolved in early vertebrates.

The crosstalk between the immune system and microbiota drives an amazingly complex mutualistic symbiosis. In mammals, the upper respiratory tract acts as a gateway for pathogen invasion, and the dynamic interaction between microbiota and mucosal immunity on its surface can effectively prevent disease development. However, the relationship between virus-mediated mucosal immune responses and microbes in lower vertebrates remains uncharacterized. In this study, we successfully constructed an infection model by intraperitoneally injecting common carp ( Cyprinus carpio ) with spring viremia of carp virus (SVCV). In addition to the detection of the SVCV in the nose and pharynx of common carp, we also identified obvious histopathological changes following viral infection. Moreover, numerous immune-related genes were significantly upregulated in the nose and pharynx at the peak of SVCV infection, after which the expression levels decreased to levels similar to those of the control group. Transcriptome sequencing results revealed that pathways associated with bacterial infection in the Toll-like receptor pathway and the Nod-like receptor pathway were activated in addition to the virus-related Rig-I-like receptor pathway after SVCV infection, suggesting that viral infection may be followed by opportunistic bacterial infection in these mucosal tissues. Using 16S rRNA gene sequencing, we further identified an upward trend in pathogenic bacteria on the mucosal surface of the nose and pharynx 4 days after SVCV infection, after which these tissues eventually reached new homeostasis. Taken together, our results suggest that the dynamic interaction between mucosal immunity and microbiota promotes the host to a new ecological state.

In this study, we aimed to characterize the fish community structure and identify the drivers contributing to homogenization/differentiation processes in four tributaries to the Pearl River, Guangxi Province, China, over the past few decades. We sampled 22 sites seasonally from 2013 through 2015, and these sites were selected based on archived records of previous sampling conducted in the 1980s. Jaccard's faunal similarity index, cluster analysis, and canonical correspondence analysis (CCA) were applied to describe the homogenization/differentiation of fish community and illustrate the potential effectors. The number of fish species present in three of the four sampled tributaries declined dramatically over the past 30 years, leading toward a trend of increased fish community homogeneity throughout the watershed. Results from multidimensional scaling and cluster analyses allowed us to divide the study area into two distinct ecoregions. Four species (yellow catfish Pelteobagrus fulvidraco, pond loach Misgurnus anguillicaudatus, Nile tilapia Oreochromis niloticus, and sharpbelly Hemiculter leucisculus) were considered to be indicative fish species contributing more than 5% of the dissimilarity between the two eco‐regions according to the results of similarity percentage procedure. Results from CCA revealed that pH and latitude corresponded with the dominant fish species of each respective tributary. More specifically, CCA results allowed us to classify dominant fish species into three distinct groups. The first group was mainly located in Guijiang characterized by higher latitudes and lower pH values, the second group was widespread in the four tributaries, and the last group was primarily distributed in Yujiang, Youjiang, and Zuojiang characterized by lower latitudes and higher pH values. Spatial differentiation of fish community structure and temporal homogeneity of species composition were attributed to the joint actions of human interventions including construction of dams and introductions of exotic fish species that led to habitat degeneration and fragmentation, and unequal interspecies competitions. The spatial differentiation of fish community structure and temporal homogeneity of species composition were attributed to the joint actions of human intervention of dam construction and exotic fish species introduction.

In our previous studies, we identified miR-16 as being downregulated during activation of hepatic stellate cells (HSCs) by microarray hybridization. However, the roles and related mechanisms of miR-16 in HSCs are not understood. In this study, The miRNA RNAi technique was used to analyze the effects of miR-16 on biological properties of HSCs in vitro. The lentiviral vector encoding miR-16 was constructed and transfected. Furthermore, the expression level of miR-16 was measured by real-time PCR. Cellular growth and proliferation capacity were assayed using the cell counting kit-8 (CCK-8). The apoptosis rate and cell-cycle distribution were measured by flow cytometry. Cell morphological characteristics were identified by phase-contrast microscopy, fluorescence microscopy and electron microscopy. The underlying mechanisms related to the changes in biological properties were assessed. The identity of the recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. Virus titer was 10 8  > ifu/m. Restoring the intracellular miRNAs by miR-16 administration greatly reduced the expression levels of cyclin D1 (CD1). Cell-cycle arrest and typical features of apoptosis were detected in activated HSCs treated with pLV-miR-16. Our results indicate that transduction of miR-16 offers a feasible approach to significantly inhibit HSC proliferation and increase the apoptosis index. Thus, targeted transfer of miR-16 into HSC may be useful for the treatment of hepatic fibrosis.

Background/aims Fibrosis occurs in most chronic liver injuries and results from changes in the balance between synthesis and degradation of extracellular matrix (ECM) components. Matrix metalloproteinases (MMPs) and their endogenous inhibitors (TIMPs) are known to regulate the ECM turnover. We investigate the effect of modified synthetic small interfering RNA (siRNA) targeting TIMP‐2 in rat model of liver fibrosis. Methods Rat hepatic fibrosis was induced by CCl4 for 8 weeks. After the 2‐week CCl4 injection period, rats in the three siRNA groups simultaneously received a different dosage (0.05, 0.1 and 0.2 mg·kg−1, respectively) of modified synthetic siRNA targeting TIMP‐2 via the tail vein every 3 days for 6 weeks. The pathological changes in liver tissues were observed by light microscopy and transmission electron microscopy. Portal vein pressure and proliferating cell nuclear antigen were measured. Expression of TIMP‐2, MMP‐2, MT1‐MMP, MMP‐13, hepatocyte growth factor, collagen type I, collagen type III and α‐SMA were evaluated by quantitative real‐time polymerase chain reaction or Western blotting or gelatin zymography. Results Modified synthetic siRNA targeting TIMP‐2 induced a dose‐dependent inhibition of the TIMP‐2 expression in the rat model of liver fibrosis with a similar trend in MMP‐2 and MT1‐MMP, but an increase in MMP‐13. Rats administered siRNA targeting TIMP‐2 showed promotion of ECM degradation, reduction in activated hepatic stellate cells and enhancement of hepatocyte regeneration. Furthermore, portal hypertension was also ameliorated after treatment with siRNA targeting TIMP‐2. Conclusions Knock‐down of TIMP‐2 expression attenuates CCl4‐induced liver fibrosis and is a potential pharmacological target for gene therapy in liver fibrosis. Copyright © 2007 John Wiley & Sons, Ltd.

The polyaniline/Mn 0.8 Zn 0.2 Fe 2 O 4 (PANI/MZF) nanocomposite was prepared by an in situ polymerization method. The samples were characterized by Fourier transform infrared spectrometer, X-ray diffraction, scanning electron microscope and vibrating sample magnetometer. The complex permittivity and complex permeability for the nanocomposites were measured by wave-guide method with vector network analyzer in 2.0–18.0 GHz. The reflection losses ( R L ) of the nanocomposites were investigated according to the wave transmission theory. The results showed the maximum reflection loss of the PANI/MZF nanocomposite was about −20.6 dB at 14.4 GHz with a bandwidth of 5.6 GHz. In conclusion, a wider absorption frequency range could be obtained by adding polyaniline contain in the MZF ferrite. The PANI/MZF nanocomposite is a good microwave shielding and absorbing materials at higher frequency.

Purpose This study aimed to determine whether functional gastrointestinal disorders are more common among adolescents with self-reported poor sleep. Methods Junior middle school and senior high school students ( n  = 1,362) were recruited from schools in Shanghai. Students completed two questionnaires: the questionnaire for irritable bowel syndrome (IBS) in adolescents and the Pittsburgh Sleep Quality Index. Results The prevalence of poor sleep was 34.29% [95% confidence interval (CI) = 31.77–36.81] and there was no significant difference between genders ( P  = 0.991). The tendency towards poor sleep increased with age, with age group yielding a significant effect ( P =  0.001). In junior middle school and senior high school students, the propensity towards poor sleep was 30.10% (95% CI = 27.08–33.12%) and 42.11% (95% CI = 37.67–46.55%), respectively. Among students with poor sleep, the prevalence of IBS was 19.70% (95% CI = 16.09–23.31). After adjusting for age, sex, night pain, and psychological factors, IBS was significantly more common in students with poor sleep (odds ratio = 1.92; 95% CI = 1.07–2.58). Conclusion We conclude that IBS is prevalent in students with poor sleep. Poor sleep was independently associated with IBS among adolescents in Shanghai China.

The skin of vertebrates is the outermost organ of the body and serves as the first line of defense against external aggressions. In contrast to mammalian skin, that of teleost fish lacks keratinization and has evolved to operate as a mucosal surface containing a skin-associated lymphoid tissue (SALT). Thus far, IgT representing the prevalent immunoglobulin (Ig) in SALT have only been reported upon infection with a parasite. However, very little is known about the types of B cells and Igs responding to bacterial infection in the teleost skin mucosa, as well as the inductive or effector role of the SALT in such responses. To address these questions, here we analyzed the immune response of trout skin upon infection with one of the most widespread fish skin bacterial pathogens, Flavobacterium columnare . This pathogen induced strong skin innate immune and inflammatory responses at the initial phases of infection. More critically, we found that the skin mucus of fish having survived the infection contained significant IgT-but not IgM- or IgD-specific titers against the bacteria. Moreover, we d emonstrate the local proliferation and production of IgT+ B-cells and specific IgT titers respectively within the SALT upon bacterial infection. Thus, our findings represent the first demonstration that IgT is the main Ig isotype induced by the skin mucosa upon bacterial infection, and that because of the large surface of the skin, its SALT probably represents a prominent IgT inductive site in fish. ### Competing Interest Statement The authors have declared no competing interest.