The NF-κB/Relish Activates miR-308 to Negatively Regulate Imd Pathway Immune Signaling in Drosophila.

The strength and duration of the NF-κB signaling response must be tightly modulated to avoid an inadequate or excessive immune response. Relish, a core NF-κB transcription factor of the Drosophila Imd pathway, can control the expression of antimicrobial peptides, including Dpt and AttA, to defend against Gram-negative bacterial infections, but whether Relish may regulate miRNA expression to participate in the immune response remains unclear. In this study, taking advantage of Drosophila S2 cells and different overexpression/knockout/knockdown flies, we first found that Relish could directly activate the expression of miR-308 to negatively regulate the immune response and promote the survival of Drosophila during Enterobacter cloacae infection. Second, our results demonstrated that Relish-mediated expression of miR-308 could suppress target gene Tab2 to attenuate the Drosophila Imd pathway signal during the middle and late stages of the immune response. Third, we detected the dynamic expression patterns of Dpt, AttA, Relish, miR-308, and Tab2 in wild-type flies after E. coli infection, which further revealed that the feedback regulatory loop of Relish-miR-308-Tab2 plays a crucial role in the immune response and homeostasis maintenance of the Drosophila Imd pathway. Overall, our present study not only illustrates an important mechanism by which this Relish-miR-308-Tab2 regulatory axis can negatively control the Drosophila immune response and participate in homeostasis maintenance but also provides new insights into the dynamic regulation of the NF-κB/miRNA expression network of animal innate immunity.

Sex different effect of antibiotic and probiotic treatment on intestinal microbiota composition in chemically induced liver injury rats.

Differences in the gut microbiota and metabolic processes between males and females may explain differences in the risk of liver injury; however, the sex-specific effects of antibiotics and probiotics on these relationships are not clear. We evaluated differences in the gut microbiota and the risk of liver injury between male and female rats after the oral administration of antibiotics or probiotics followed by a period of diethylnitrosamine treatment to chemically induce liver injuryusing high-throughput sequencing of fecal microbiota combined with histological analyses of liver and colon tissues. Our results suggest that the ratio of gram-positive to gram-negative bacteria in kanamycin-treated rats was significantly higher than that of other groups, and this difference persisted for the duration of the experiment. Antibiotics significantly changed the composition of the gut microbiota of experimental rats. Clindamycin caused more diethylnitrosamine-induced damage to livers of male rats. Probiotics did not influencethe gut microbiota; however, they hadprotective effects against liver injury induced by diethylnitrosamine, especially in female rats. These results strengthen our understanding of sex differences in the indirect effects of antibiotics or probiotics on metabolism and liver injury in hosts via the gut microbiota.

Complete genome analysis of Pseudomonas furukawaii ZS1 isolated from grass carp (Ctenopharyngodon idellus) culture water.

Pseudomonas furukawaii ZS1, isolated from grass carp (Ctenopharyngodon idellus) culture water, exhibits efficient aerobic nitrate reduction without nitrite accumulation; however, the molecular pathway underlying this aerobic nitrate reduction remains unclear. In this study, we constructed a complete genome map of P. furukawaii ZS1 and performed a comparative genomic analysis with a reference strain. The results showed that P. furukawaii ZS1 genome was 6 026 050 bp in size and contained 5427 predicted protein-coding sequences. The genome contained all the necessary genes for the dissimilatory nitrate reduction to ammonia pathway but lacked those for the assimilatory nitrate reduction pathway; additionally, genes that convert ammonia to organic nitrogen were also identified. The presence of putative genes associated with the nitrogen and oxidative phosphorylation pathways implied that ZS1 can perform respiration and nitrate reduction simultaneously under aerobic conditions, so that nitrite is rapidly consumed for detoxication by denitrification. The aim of this study is to indicate the great potential of strain ZS1 for future full-scale applications in aquaculture. This work provided insights at the molecular level on the nitrogen metabolic pathways in Pseudomonas species. The understanding of nitrogen metabolic pathways also provides significant molecular information for further Pseudomonas species modification and development.

Effects of organic and inorganic nitrogen sources on the transcriptome of gellan gum biosynthesis by Sphingomonas paucimobilis.

AIM: Investigate the effects of different nitrogen sources on the metabolic characteristics of Sphingomonas paucimobilis during gellan gum (GG) production was helpful for developing optimized conditions that are widely applicable to all GG production processes. METHODS AND RESULTS: We compared the effects of organic nitrogen (ON) and inorganic nitrogen (IN) sources during GG production using transcriptome sequencing. Our results showed that compared with the IN source, the ON source effectively improved the cell number and GG production of S. paucimobilis during fermentation. There were significant differences in gene transcription levels between the ON and IN groups at different fermentation times. CONCLUSIONS: The transcriptional levels of multiple genes in the pathways from α-D-glucose-1P to glyceraldehyde-3P were reduced in the ON group, whereas those of multiple genes in the pathways from glyceraldehyde-3P to acetyl-CoA were significantly enhanced in the ON group after 12 h of fermentation. The transcription levels of multiple genes participating in the citrate cycle and upstream of fatty acid metabolism pathways were significantly enhanced in the ON group after 12 h of fermentation. Except for the transcripts per million (TPMs) of pgm and rfbA genes in ON, which were significantly higher than those in IN at 12 h after fermentation, the TPMs of the majority of genes in ON were significantly lower than those in IN. The transcription levels of genes participating in the transformation of N-acetyl-D-glucosamine (GlcNAc) to UDP-N-acetyl-alpha-D-glucosamine (UDP-GlcNAc) were enhanced in the ON group during the fermentation process.

Presence of TSHR in NK Cells and Action of TSH on NK Cells.

INTRODUCTION: Thyroid-stimulating hormone receptor (TSHR) is widely expressed in human tissues and cells. TSHR is not only involved in thyroid disease but also in the neuroendocrine-immune regulatory network. However, no study has exclusively focused on the expression and function of TSHR in natural killer (NK) cells. METHODS: We studied TSHR expression using reverse transcription PCR to verify TSHR mRNA transcripts in human and mouse NK cells. Human and mouse thyroid and liver tissues as well as peripheral blood mononuclear cells (PBMCs) or spleen lymphoid cells (SLCs) were used as controls. The TSHR protein levels in NK-92 cells were determined by immunofluorescence staining. The function of TSHR in NK cells was investigated by measuring the TSH-stimulated cAMP levels. RESULTS: TSHR mRNA was detected in human and mouse NK cells as well as in NK-92 cells and had the same sequence as that of thyroid-derived, PBMC-derived, and liver-derived mRNA. The TSHR protein was also expressed in the cell membrane of NK-92 cells. Furthermore, the cAMP levels in NK-92 cells were significantly higher after adding 102 mIU/mL of bovine TSH at p < 0.05, which stimulated cAMP production in NK-92 cells. CONCLUSIONS: Our findings confirm that TSHR is present and functional in NK cells and provide key clues for the potential regulatory effects of TSH on TSHR in NK cells in the immune system.

DNL-Net: deformed non-local neural network for blood vessel segmentation.

BACKGROUND: The non-local module has been primarily used in literature to capturing long-range dependencies. However, it suffers from prohibitive computational complexity and lacks the interactions among positions across the channels. METHODS: We present a deformed non-local neural network (DNL-Net) for medical image segmentation, which has two prominent components; deformed non-local module (DNL) and multi-scale feature fusion. The former optimizes the structure of the non-local block (NL), hence, reduces the problem of excessive computation and memory usage, significantly. The latter is derived from the attention mechanisms to fuse the features of different levels and improve the ability to exchange information across channels. In addition, we introduce a residual squeeze and excitation pyramid pooling (RSEP) module that is like spatial pyramid pooling to effectively resample the features at different scales and improve the network receptive field. RESULTS: The proposed method achieved 96.63% and 92.93% for Dice coefficient and mean intersection over union, respectively, on the intracranial blood vessel dataset. Also, DNL-Net attained 86.64%, 96.10%, and 98.37% for sensitivity, accuracy and area under receiver operation characteristic curve, respectively, on the DRIVE dataset. CONCLUSIONS: The overall performance of DNL-Net outperforms other current state-of-the-art vessel segmentation methods, which indicates that the proposed network is more suitable for blood vessel segmentation, and is of great clinical significance.

Comparative proteomic analysis of children FSGS FFPE tissues.

BACKGROUND: In children, focal segmental glomerulosclerosis (FSGS) is the main cause of steroid resistant nephrotic syndrome (SRNS). To identify specific candidates and the mechanism of steroid resistance, we examined the formalin-fixed paraffin embedded (FFPE) renal tissue protein profiles via liquid chromatography tandem mass spectrometry (LC-MS/MS). METHODS: Renal biopsies from seven steroid-sensitive (SS) and eleven steroid-resistant (SR) children FSGS patients were obtained. We examined the formalin-fixed paraffin embedded (FFPE) renal tissue protein profiles via liquid chromatography tandem mass spectrometry (LC-MS/MS). Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment and Gene Ontology (GO) analysis, as well as the construction of protein-protein interaction (PPI) network were performed. Two proteins were further valiadated by immunohistochemistry staining in FSGS patients and mice models. RESULTS: In total, we quantified more than 4000 proteins, of which 325 were found to be differentially expressed proteins (DEPs) between the SS and SR group (foldchange ≥2, P<0.05). The results of GO revealed that the most significant up-regulated proteins were primarily related to protein transportation, regulation of the complement activation process and cytolysis. Moreover, clustering analysis showed differences in the pathways (lysosome, terminal pathway of complement) between the two groups. Among these potential candidates, validation analyses for LAMP1 and ACSL4 were conducted. LAMP1 was observed to have a higher expression in glomerulus, while ACSL4 was expressed more in tubular epithelial cells. CONCLUSIONS: In this study, the potential mechanism and candidates related to steroid resistance in children FSGS patients were identified. It could be helpful in identifying potential therapeutic targets and predicting outcomes with these proteomic changes for children FSGS patients.

High prevalence of thyroid hormone autoantibody and low rate of thyroid hormone detection interference.

OBJECTIVE: Thyroid hormone autoantibody (THAb) is a common antibody in autoimmune disease and can interfere with the detection of thyroid hormone (TH). There was no research reporting the prevalence of THAb in Chinese and the rate of THAb interfering with TH detection. METHODS: We collected 114 patients with autoimmune thyroid disease (AITD) (Hashimoto's thyroiditis, 57 cases; Graves' disease, 57 cases), 106 patients with nonthyroid autoimmune diseases (NTAID), and 120 healthy subjects. According to the presence or absence of thyroid antibodies, patients with NTAID were divided into two groups: NTAID-AITD and NTAID groups. Radioimmunoprecipitation technique was used to detect THAb in all subjects. TH was detected on Abbot and Roche platforms in patients with positive THAb. RESULTS: The prevalence of THAb was 22.8% in Hashimoto's thyroiditis and 45.6% in Graves' disease. The prevalence of THAb in AITD group was lower than that in NTAID or NTAID-AITD groups (34.2% vs. 61.5%, p = 0.014; 34.2% vs. 71.3%, p < 0.01). Among total 98 patients with positive THAb, TH levels of 9 patients were falsely elevated (9.18%). CONCLUSION: The prevalence of THAb in AITD patients was lower than that in NTAID patients. Although THAb had a high frequency in various autoimmune diseases, the prevalence of THAb interfering with TH detection was only 9.18%.

Associations Between Cessation of Second-Line Therapies and Relapse Rates of Childhood Refractory Minimal-Change Nephrotic Syndrome: A Single-Center, Retrospective Chart Review.

Background: Most patients (≥85%) with minimal-change nephrotic syndrome (MCNS) respond to corticosteroid treatment. However, about 10% to 20% of patients with MCNS have steroid-resistant nephrotic syndrome and 25% to 43% of patients have steroid-dependent nephrotic syndrome or frequent-relapse steroid-sensitive nephrotic syndrome. Patients with refractory MCNS are treated with various second-line therapies. Objectives: This study aimed to evaluate the associations between the use of various second-line therapies and relapse rates in Chinese patients with childhood refractory MCNS. Methods: In this study, patients with childhood nephrotic syndrome renal biopsy proved to be "minimal change" from a single tertiary-care center between January 2002 and July 2018 were identified. A Total of 56 medical charts of patients treated with 1 of these second-line immunosuppressors: cyclophosphamide (CYC), mycophenolate mofetil (MMF), or tacrolimus (TAC) were reviewed. Patients were divided into CYC (n = 24), MMF (n = 20), and TAC (n = 12) groups according to the second-line therapy administered. Baseline characteristics, immune status, immunocomplex deposition in the renal tissue, and treatment outcomes were analyzed. Results: The ratio of patients with steroid-resistant nephrotic syndrome and steroid-dependent nephrotic syndrome in the CYC, MMF, and TAC groups did not differ significantly (P = 0.721). The immunofluorescence assay did not show any significant differences in immunocomplex deposition identified in renal biopsy specimens among the 3 groups. The rate of steroid-free remission in the TAC group (75%) was higher than that in the MMF (55%) and CYC (25%) groups (P = 0.012). At the last follow-up, two-thirds of children in the TAC group had a relapse following discontinuation of therapy. In the TAC group, patients for whom steroids were withdrawn had significantly higher levels of immunoglobulin G at the onset of nephrotic syndrome than those for whom steroids were continued (P = 0.017). In the MMF group, children with relapse had a significantly higher percentage of CD16+CD56+-positive cells than those without relapse (P = 0.042). The relapse rate after treatment discontinuation was significantly different among the 3 groups (P = 0.035). Notably, the relapse rate after treatment discontinuation in the CYC group was lower than those in the other 2 groups (P = 0.035). Conclusions: In this small population of Chinese patients with childhood refractory MCNS, the relapse rate following TAC therapy was higher than that following MMF or CYC therapy. Different proportions of CD16+CD56+-positive cells might be associated with relapse rates in patients with MCNS receiving MMF treatment. (Curr Ther Res Clin Exp. 2022; 83:XXX-XXX).

Seasonal variation significantly affected bacterioplankton and eukaryoplankton community composition in Xijiang River, China.

Both bacterioplankton and eukaryoplankton communities play important roles in the geochemical cycles and energy flows of river ecosystems. However, whether a seasonal change in bacterioplankton and eukaryoplankton communities is synchronous remains unclear. To test the synchronicity and analyze how physical and chemical environmental factors affect these communities, we compared bacterioplankton and eukaryoplankton communities in surface water samples between March (dry season) and June (rainfall season) considering water environmental factors. Our results showed that there was no significant difference in operational taxonomic unit number, Shannon index, and Chao1 index in bacterioplankton and eukaryoplankton communities between March and June. However, principal component analysis showed that the communities were significantly different between the sampling times and sampling sites. Water temperature (WT), oxidation-reduction potential (ORP), water transparency (SD), NO3-N, and NH3 significantly influenced bacterioplankton communities, and WT, SD, ORP, and NH4-N significantly influenced eukaryoplankton communities in the river. These results implied that compared with the sampling sites, sampling times more significantly affected the bacterioplankton and eukaryoplankton river communities by influencing WT, ORP, SD, and nitrogen forms.

MicroRNA miR-425 inhibits proliferation and migration of colorectal cancer HCT116 cells.

OBJECTIVE: To assess the role of micro ribonucleic acid 425 in colorectal carcinogenesis. METHODS: The experimental study was conducted from December 2016 to July 2017 at the Xinjiang Dingju Medical Laboratory, China, and comprised sphere formation assay, wound healing assay, transwell assay, thiazolyl blue tetrazolium bromide cell proliferation assay, flow cytometric analysis, reverse transcription quantitative polymerase chain reaction and western blotting to analyse the proliferation and invasion capability of HCT116 cells transfected with a micro ribonucleic acid-425 mimic, micro ribonucleic acid-425 inhibitor, micro ribonucleic acid-425 mimic negative control, and micro ribonucleic acid-425 inhibitor negative control. RESULTS: Micro ribonucleic acid-425 expression in HCT116 cells was up regulated after transfection, resulting in inhibition of sphere formation. Over-expression of micro ribonucleic acid-425 inhibited the proliferation of HCT116 cells and induced apoptosis along with inhibition of HCT116 cell migration and invasion. CONCLUSION: Over-expression of micro ribonucleic acid-425 was found to have the potential to inhibit sphere formation as well as migration and invasion of HCT116 cells by inhibiting proliferation and promoting apoptosis.

DjERas plays an important role in planarian regeneration and homeostasis.

The mechanisms of cell turnover including cell proliferation and cell differentiation were complex. Planarians possess amazing regeneration ability and undergo cell turnover throughout life. We identified a homologous gene of ERas by RNAi in Dugesia japonica. Knocking-down DjERas resulted in regeneration and homeostasis defects. Furthermore, we found that the expression of neoblasts and late progeny marker gene decreased in DjERas RNAi planarians. Our studies indicated that down-regulation of DjERas inhibited the proliferation and differentiation of stem cells through the conserved signaling pathway, resulted in the inability of the planarian to regenerate and maintain homeostasis. Our results suggest that DjERas plays a crucial role in the process of cell turnover.

Effect of green tea and mulberry leaf powders on the gut microbiota of chicken.

BACKGROUND: The gut microbiota is closely correlated with host health and is strongly influenced by food composition. Chinese herbs are usually used as natural feed additives in livestock production. Therefore, the present study assessed the influence of diet supplementation with green tea and mulberry leaf powders on the chicken gut microbiota. The gut microbiota compositions were determined using 16S rDNA sequencing. RESULTS: Enhanced relative abundances of Bacteroides, Prevotella, and Megamonas were found in the chicken gut when mulberry leaf powder was added to diet. Conversely, a higher abundance of potentially pathogenic Gallibacterium was found in the chicken gut when the diet was supplemented with green tea powder. These results indicated that green tea powder and mulberry leaf powder can greatly affect the gut microbiota of chickens by changing their compositions. CONCLUSIONS: It is imperative to examine and evaluate the effects of Chinese herbs on animal health before they are introduced as feed additives in animal production.

MicroRNA-709 Mediates Acute Tubular Injury through Effects on Mitochondrial Function.

Mitochondrial dysfunction has important roles in the pathogenesis of AKI, yet therapeutic approaches to improve mitochondrial function remain limited. In this study, we investigated the pathogenic role of microRNA-709 (miR-709) in mediating mitochondrial impairment and tubular cell death in AKI. In a cisplatin-induced AKI mouse model and in biopsy samples of human AKI kidney tissue, miR-709 was significantly upregulated in the proximal tubular cells (PTCs). The expression of miR-709 in the renal PTCs of patients with AKI correlated with the severity of kidney injury. In cultured mouse PTCs, overexpression of miR-709 markedly induced mitochondrial dysfunction and cell apoptosis, and inhibition of miR-709 ameliorated cisplatin-induced mitochondrial dysfunction and cell injury. Further analyses showed that mitochondrial transcriptional factor A (TFAM) is a target gene of miR-709, and genetic restoration of TFAM attenuated mitochondrial dysfunction and cell injury induced by cisplatin or miR-709 overexpression in vitro Moreover, antagonizing miR-709 with an miR-709 antagomir dramatically attenuated cisplatin-induced kidney injury and mitochondrial dysfunction in mice. Collectively, our results suggest that miR-709 has an important role in mediating cisplatin-induced AKI via negative regulation of TFAM and subsequent mitochondrial dysfunction. These findings reveal a pathogenic role of miR-709 in acute tubular injury and suggest a novel target for the treatment of AKI.

MicroRNA-30e targets BNIP3L to protect against aldosterone-induced podocyte apoptosis and mitochondrial dysfunction.

MicroRNAs are essential for the maintenance of podocyte homeostasis. Emerging evidence has demonstrated a protective role of microRNA-30a (miR-30a), a member of the miR-30 family, in podocyte injury. However, the roles of other miR-30 family members in podocyte injury are unclear. The present study was undertaken to investigate the contribution of miR-30e to the pathogenesis of podocyte injury induced by aldosterone (Aldo), as well as the underlying mechanism. After Aldo treatment, miR-30e was reduced in a dose-and time-dependent manner. Notably, overexpression of miR-30e markedly attenuated Aldo-induced apoptosis in podocytes. In agreement with this finding, miR-30e silencing led to significant podocyte apoptosis. Mitochondrial dysfunction (MtD) has been shown to be an early event in Aldo-induced podocyte injury. Here we found that overexpression of miR-30e improved Aldo-induced MtD while miR-30e silencing resulted in MtD. Next, we found that miR-30e could directly target the BCL2/adenovirus E1B-interacting protein 3-like (BNIP3L) gene. Aldo markedly enhanced BNIP3L expression in podocytes, and silencing of BNIP3L largely abolished Aldo-induced MtD and cell apoptosis. On the contrary, overexpression of BNIP3L induced MtD and apoptosis in podocytes. Together, these findings demonstrate that miR-30e protects mitochondria and podocytes from Aldo challenge by targeting BNIP3L.

Down-regulate of Djrfc2 causes tissues hypertrophy during planarian regeneration.

Planarians are an ideal model organism for regeneration research due to their amazing ability to regenerate. DNA replication is crucial for genome stability. Replication factor C (RFC), which is a replication factor C-like complex and plays an important role during DNA replication in eukaryotes, has been reported as a wound response factor during planarian regeneration. However, how RFC controls regeneration in planarians by regulating DNA replication remains to be explained. Here, we used a two-dimensional electrophoresis (2-DE) proteomic approach to identify differentially expressed proteins in intact and regenerated planarians. Approximately 132 protein spots showed differences between intact and regenerative tissues. We selected 21 significantly expressed protein spots and processed them using TOF MS analysis. Finally, we cloned three of these candidate genes (Djhsp70, Djrfc2, Djfaim), focusing on the function of Djrfc2 during regeneration. We found that the distribution of Djrfc2 tends toward the wound site. RNA interference (RNAi) of Djrfc2 increases the number of dividing cells and the expression level of planarian neoblast marker genes, which may result in hyper-proliferation. Our studies use an available approach to directly study the regeneration dynamic at the protein level and provide further evidence to support a function of Djrfc2 in planarian regeneration.

Nearly a decade-long repeatable seasonal diversity patterns of bacterioplankton communities in the eutrophic Lake Donghu (Wuhan, China).

Uncovering which environmental factors govern community diversity patterns and how ecological processes drive community turnover are key questions related to understand the community assembly. However, the ecological mechanisms regulating long-term variations of bacterioplankton communities in lake ecosystems remain poorly understood. Here we present nearly a decade-long study of bacterioplankton communities from the eutrophic Lake Donghu (Wuhan, China) using 16S rRNA gene amplicon sequencing with MiSeq platform. We found strong repeatable seasonal diversity patterns in terms of both common (detected in more than 50% samples) and dominant (relative abundance >1%) bacterial taxa turnover. Moreover, community composition tracked the seasonal temperature gradient, indicating that temperature is a key environmental factor controlling observed diversity patterns. Total phosphorus also contributed significantly to the seasonal shifts in bacterioplankton composition. However, any spatial pattern of bacterioplankton communities across the main lake areas within season was overwhelmed by their temporal variabilities. Phylogenetic analysis further indicated that 75%-82% of community turnover was governed by homogeneous selection due to consistent environmental conditions within seasons, suggesting that the microbial communities in Lake Donghu are mainly controlled by niche-based processes. Therefore, dominant niches available within seasons might be occupied by similar combinations of bacterial taxa with modest dispersal rates throughout different lake areas.

Composition of Gut Microbiota in the Gibel Carp (Carassius auratus gibelio) Varies with Host Development.

To understand how a bacteria-free fish gut ecosystem develops microbiota as the fish ages, we performed a 1-year study on the gut microbiota of hatchling gibel carp (Carassius auratus gibelio). Our results indicate that the gut microbial diversity increases significantly as the fish develop. The gut microbial community composition showed significant shifts corresponding to host age and appeared to shift at two time points despite consistent diet and environmental conditions, suggesting that some features of the gut microbial community may be determined by the host's development. Dietary and environmental changes also seem to cause significant shifts in the fish gut microbial community. This study revealed that the gut microbiota of gibel carp assemble into distinct communities at different times during the host's development and that this process is less affected by the surrounding environment than by the host diet and development. Community phylogenetic analyses based on the net relatedness index further showed that environmental filtering (host selection) deterministically governs the gut microbial community composition. More importantly, the influence of host-associated deterministic filtering tends to weaken significantly over the course of the host's development. However, further studies are needed to assess whether this host development-dependent shift in gut microbiota will still exist under different rearing strategies.

Environmental filtering decreases with fish development for the assembly of gut microbiota.

Gut microbiota typically occupy habitats with definable limits/borders that are comparable to oceanic islands. The gut therefore can be regarded as an 'island' for the assembly of microbial communities within the 'sea' of surrounding environments. This study aims to reveal the ecological mechanisms that govern microbiota in the fish gut 'island' ecosystem. Taxonomic compositions, phylogenetic diversity, and community turnover across host development were analyzed via the high-throughput sequencing of 16S rRNA gene amplicons. The results indicate that the Shannon diversity of gut microbiota in the three examined freshwater fish species all significantly decreased with host development, and the dominant bacterial taxa also changed significantly during host development. Null model and phylogenetic-based mean nearest taxon distance (MNTD) analyses suggest that host gut environmental filtering led to the assembly of microbial communities in the fish gut 'island'. However, the phylogenetic clustering of local communities and deterministic processes that governed community turnover became less distinct as the fish developed. The observed mechanisms that shaped fish gut microbiota seemed to be mainly shaped by the gut environment and by some other selective changes accompanying the host development process. These findings greatly enhance our understanding of stage-specific community assembly patterns in the fish gut ecosystem.

NLRP3 inflammasome mediates albumin-induced renal tubular injury through impaired mitochondrial function.

Proteinuria serves as a direct causative factor of renal tubular cell injury and is highly associated with the progression of chronic kidney disease via uncertain mechanisms. Recently, evidence demonstrated that both NLRP3 inflammasome and mitochondria are involved in the chronic kidney disease progression. The present study was undertaken to examine the role of NLRP3 inflammasome/mitochondria axis in albumin-induced renal tubular injury. In patients with proteinuria, NLRP3 was significantly up-regulated in tubular epithelial cells and was positively correlated with the severity of proteinuria. In agreement with these results, albumin remarkably activated NLRP3 inflammasome in both in vitro renal tubular cells and in vivo kidneys in parallel with significant epithelial cell phenotypic alteration and cell apoptosis. Genetic disruption of NLRP3 inflammasome remarkably attenuated albumin-induced cell apoptosis and phenotypic changes under both in vitro and in vivo conditions. In addition, albumin treatment resulted in a significant mitochondrial abnormality as evidenced by the impaired function and morphology, which was markedly reversed by invalidation of NLRP3/caspase-1 signaling pathway. Interestingly, protection of mitochondria function by Mn(III)tetrakis (4-benzoic acid) porphyrin (MnTBAP) or cyclosporin A (CsA) robustly attenuated albumin-induced injury in mouse proximal tubular cells. Collectively, these findings demonstrated a pathogenic role of NLRP3 inflammasome/caspase-1/mitochondria axis in mediating albumin-induced renal tubular injury. The discovery of this novel axis provides some potential targets for the treatment of proteinuria-associated renal injury.