Sex-bias of core intestinal microbiota in different stocks of Chinese mitten crabs (Eriocheir sinensis).

The differences in intestinal microbiota composition are synergistically shaped by internal and external factors of the host. The core microbiota plays a vital role in maintaining intestinal homeostasis. In this study, we conducted 16S rRNA sequencing analysis to investigate the stability of intestinal microbiota and sex-bias of six stocks of Chinese mitten crabs (105 females; and 110 males). The dominant phyla in all six stocks were Proteobacteria, Tenericutes, Bacteroidetes and Firmicutes; however, their relative abundance differed significantly. Twenty-seven core operational taxonomic units (OTUs), corresponding to 18 genera, were screened. Correlation analysis revealed that OTUs of four stocks in the Yangtze River system play important roles in maintaining the stability of intestinal microbiota. Additionally, the core intestinal microbiota was significantly sex-biased, and the top three genera in terms of relative abundance (Acinetobacter, Vibrio, and Candidatus_Hepatoplasma) were significantly dominant in female crabs. Network structure analysis also confirmed gender differences in the association pattern of intestinal microbiota. The intestinal microbiota of male crabs has a higher degree of functional enrichment. This study provided a theoretical basis for further investigating exploring the shaping effect of gender and geographical factors on the intestinal microbiota of Chinese mitten crabs.

Assessing the ecotoxicity of florfenicol exposure at environmental levels: A case study of histology, apoptosis and microbiota in hepatopancreas of Eriocheir sinensis.

The intensification of production practices in the aquaculture industry has led to the indiscriminate use of antibiotics to combat diseases and reduce costs, which has resulted in environmental pollution, posing serious threats to aquaculture sustainability and food safety. However, the toxic effect of florfenicol (FF) exposure on the hepatopancreas of crustaceans remains unclear. Herein, by employing Chinese mitten crab (Eriocheir sinensis) as subjects to investigate the toxic effects on histopathology, oxidative stress, apoptosis and microbiota of hepatopancreas under environment-relevant (0.5 and 5 µg/L), and extreme concentrations (50 µg/L) of FF. Our results revealed that the damage of hepatopancreas tissue structure caused by FF exposure in a dose-and time-dependent manner. Combined with the increased expression of apoptosis-related genes (Caspase 3, Caspase 8, p53, Bax and Bcl-2) at mRNA and protein levels, activation of catalase (CAT) and superoxide dismutase (SOD), and malondialdehyde (MDA) accumulation, FF exposure also induced oxidative stress, and apoptosis in hepatopancreas. Interestingly, 7 days exposure triggered more pronounced toxic effect in crabs than 14 days under environment-relevant FF concentration. Integrated biomarker response version 2 (IBRv2) index indicated that 14 days FF exposure under extreme concentration has serious toxicity effect on crabs. Furthermore, 14 days exposure to FF changed the diversity and composition of hepatopancreas microbiota leading remarkable increase of pathogenic microorganism Spirochaetes following exposure to 50 µg/L of FF. Taken together, our study explained potential mechanism of FF toxicity on hepatopancreas of crustaceans, and provided a reference for the concentration of FF to be used in culture of Chinese mitten crab.

Hypoxia stress induces complicated miRNA responses in the gill of Chinese mitten crab (Eriocheir sinensis).

Hypoxia caused by global climate change and human activities has become a growing concern eliciting serious damages to aquatic animals. microRNAs (miRNAs) as non-coding regulatory RNAs exert vital effects on hypoxia responses. Chinese mitten crab (Eriocheir sinensis) with the habitat on the sediment surface or the pond bottom is susceptible to oxygen deficiency. However, whether miRNAs are involved in the response of the crabs to hypoxia stress remains enigmas. In this study, we conducted the whole transcriptome-based miRNA-mRNA integrated analysis of Chinese mitten crab gill under hypoxic condition for 3 h and 24 h We found that the acute hypoxia induces complex miRNA responses with the extensive influences on their target genes that engaged in various bio-processes, especially those associated with immunity, metabolism and endocrine. The impact of hypoxia on crab miRNAs is severer, as the exposure lasts longer. In response to the dissolved oxygen fluctuation, the HIF-1 signaling is activated by miRNAs to cope with the hypoxia stress through strategies including balancing inflammatory and autophagy involved in immunity, changing metabolism to reducing energy consumption, and enhancing oxygen-carrying and delivering capacities. The miRNAs and their corresponding target genes engaged in hypoxia response were intertwined into an intricate network. Moreover, the top hub molecular, miR-998-y and miR-275-z, discovered from the network might serve as biomarkers for hypoxia response in crabs. Our study provides the first systemic miRNA profile of Chinese mitten crab induced by hypoxia stress, and the identified miRNAs and the interactive network add new insights into the mechanism of hypoxia response in crabs.

Effects of hypoxia and reoxygenation on apoptosis, oxidative stress, immune response and gut microbiota of Chinese mitten crab, Eriocheir sinensis.

Hypoxia causes irreversible damage to aquatic animals. However, few reports have explored the effect of hypoxia stress and reoxygenation on intestinal homeostatic imbalance and consequent hepatopancreas-intestine axis health in crustacean. Herein, 180 Chinese mitten crabs (Eriocheir sinensis) were equally divided into control (DO 7.0 ± 0.2 mg/L) and treatment groups. The treatment group was exposed with continuous hypoxic stress (DO 3.0 ± 0.1 mg/L) for 96 h and then reoxygenated (DO 6.9 ± 0.1 mg/L) for 96 h. The effects on intestines and hepatopancreas of Chinese mitten crab were investigated, and the role of gut microbiota in hypoxia induced damages was explored. Hypoxia impaired intestinal tissue structure, and decreased swelling and the number of goblet cells, which are features that did not significantly improve after reoxygenation. With prolonged hypoxic stress, the activities of antioxidant enzymes (LDH, SOD and CAT) and MDA content in intestine were significantly elevated. Moreover, the level of oxidative stress increased, which led to upregulated apoptosis rate and expression of apoptosis-related genes (Caspase 3, Caspase 8 and BAX). In addition, the expression of immune related genes (MyD88, ALF1, Relish and Crustin) in hepatopancreas and intestine was both significantly induced under hypoxia, which activated the immune defense mechanism of Chinese mitten crab to adapt to the hypoxic environment. Furthermore, diversity and relative abundance of gut microbiota decreased noticeably during hypoxic stress; the number of beneficial bacteria downregulated. Finally, KEGG pathway analysis revealed that nine pathways were significantly enriched in intestinal microorganisms, including autoimmune disease and environmental adaptation. Collectively, these results suggested that hypoxia negatively affected E. sinensis health by triggering oxidative stress, altering the composition of the gut microbiota and inhibiting the immune response.

Analysis of candidate biomarkers and related transcription factors involved in the development and restoration of stress-induced gastric ulcer by transcriptomics.

Stress-induced gastric ulcer is one of the common complications affecting patients after trauma, mainly leading to gastrointestinal bleeding and perforation, and severe cases may be life-threatening. However, the molecular mechanism of stress-induced gastric ulcer remains unclear. In the present study, RNA-sequencing was performed on gastric tissues of normal rats (C), stress-induced gastric ulcer rats (T0), and rats recovered from gastric ulcer for 3 days (T3), and bioinformatics analysis was performed to determine changes in gene expression and biological pathways. The protein-protein interaction (PPI) networks of differentially expressed genes (DEGs) were constructed by STRING and visualized by the Cytoscape software. The associated transcriptional factor (TFs)-gene regulatory network of the hub DEGs was also constructed. Pairwise comparisons obtained 103 (T0_C), 127 (T3_T0), and 13 (T3_C) DEGs, respectively. Gene ontology (GO) enrichment analysis indicated DEGs in T0_C and T3_T0 were significantly enriched in response to oxygen-containing compound, response to organic substance, and response to external stimulus. Pathway analysis suggested that DEGs were enriched in TNF signaling pathway, PPAR signaling pathway, apoptosis, and IL-17 signaling pathway. Seven hub genes (Fos, Jun, Nfkbia, Dusp1, Pim3, Junb, and Fosb) were obtained from the PPI networks of T0_C and T3_T0. Key TFs with close interactions, such as Fos, Jun, Nfkbia, Junb, Egr1, and Fosb, were screened This study used RNA-sequencing and bioinformatics analysis to screen out genes associated with gastric ulcer, which can help reveal the molecular mechanism of gastric ulcer development and restoration, and provide reference for the treatment of human gastric ulcers.

Predictive relevance of PD-L1 expression with pre-existing TILs in gastric cancer.

Expression of programmed cell death receptor ligand 1 (PD-L1) has been shown to be up-regulated in some gastric cancer patients and to correlate with the density of tumour infiltrating lymphocytes (TILs). However, conflicting results have been reported regarding TILs and the expression of PD-L1 as a prognostic marker for gastric cancer. We investigated the correlation of PD-L1 and TILs expression with clinicpathological characteristics in 105 well characterized gastric cancer patients. PD-L1 expression and CD3+ and CD8+ TILs were evaluated by fluorescent multiplex immunohistochemistry (mIHC) analysis. PD-L1 positive staining on tumour cells was observed in 35% cases and 48% cases showed PD-L1 expression on immune cells. Up-regulated PD-L1 expression on tumour cells and immune cells was associated with high density of pre-existing tumour infiltrating CD3+ and CD8+. In additional, more than 70% tumor infiltrating CD3+ cells were CD3+CD8+ cells. More than 60% PD-L1+ immune cells were PD-L1+CD3+CD8+ cells. PD-L1 expression in tumour cells was associated with poor prognosis and high density CD3+ and CD8+ TILs indicated improved overall survival in gastric cancer patients. Increased PD-L1 expression with low density CD3+ and CD8+ TILs had the shortest overall survival. In accordingly, PD-L1 absence with high density CD3+ and CD8+ TILs indicated the best prognosis. Combination of PD-L1 with pre-existing TILs may be more precise than PD-L1 alone for predicting survival in gastric cancer.

RISK OF SILICONE OIL AS VITREOUS TAMPONADE IN PARS PLANA VITRECTOMY: A Systematic Review and Meta-Analysis.

PURPOSE: The authors examined the differences between silicone oil and other vitreous tamponades or placebo in performing pars plana vitrectomy. METHODS: This review and meta-analysis was conducted in accordance with the PRISMA guidelines. Seven databases and the reference lists of the retrieved randomized controlled trial articles were searched to identify eligible studies. The primary outcomes were the rate of redetachment after endotamponade removal, the rate of reoperation, and poor visual acuity. The secondary outcomes were adverse events and quality of life related to postoperative position. RESULTS: Ten articles (12 trials) were included. There were no significant differences between silicone oil and other agents in most of the primary and second outcomes. Only the risk of hypotony was found to be significantly lower when filling with silicone oil, compared with other agents. No trial reported the quality of life related to postoperative position. CONCLUSION: Based on the available studies, the authors conclude that there is no significant difference in the risk of poor outcomes between pars plana vitrectomy with silicone oil and that with other vitreous tamponades with different surgical histories.

The efficacy and safety of a novel posterior scleral reinforcement device in rabbits.

PURPOSE: To evaluate the efficacy and safety of posterior scleral reinforcement (PSR) device for myopia suppression in rabbits' eyes. METHODS: PSR surgery was performed on the normal 12 8-week-old New Zealand white rabbits' right eyes. To determine efficacy of the device, ophthalmic examination would be taken at pre-operation and post-operation (1 week, 1 month, 3 months, 6 months, and 1 year), such as A-ultrasound, diopter and B-ultrasound. Evaluation of safety were based on the following indicators: intraocular pressure (IOP), slit lamp, fundus photography, fundus fluorescein angiography and pathological examination after surgery. The efficacy and safety of PSR device were evaluated by comparison (treated eyes and contralateral eyes) of pre and post-operation. RESULTS: The novel PSR device could significantly shorten axial length (preoperative axial length: 16.36 ± 0.14 mm, postoperative 1 week, 1 month, 3 months, 6 months and 1 year axial lengths: 15.03 ± 0.28 mm, 15.23 ± 0.32 mm, 15.39 ± 0.31 mm, 15.45 ± 0.22 mm and 15.45 ± 0.22 mm; P=0.00037<0.001) in the treated eyes (right eyes) after surgery. At different postoperative time points, the B-ultrasound images showed that the PSR located in appropriate position and supported the posterior sclera very well. At the same time, IOP of treated eyes kept a relatively stable level (preoperative IOP: 12.56 ± 2.01 mmHg, postoperative IOP: ranging from 11.33 ± 1.23 mmHg to 13.44 ± 2.19 mmHg, P>0.05) post-operation 1 year. During observation period, there was no significant inflammatory reaction and complications such as anterior chamber flare, empyema, endophthalmitis, vitreous hemorrhage, retina detachment and retinal choroid neovascularization by slit lamp, fundus photography and fundus fluorescein angiography. In addition, there were no pathologic changes be found by comparison treated eyes group and contralateral group eyes based on pathological examinations. CONCLUSIONS: In vivo study, effectively and safely, the novel PSR device can inhibit rabbits' axial length elongation during postoperative 1 year. This study demonstrates that this novel PSR could be a potential treatment approach for myopia.

Long-term effects of short hairpin RNA-targeted human telomerase reverse transcriptase on suppression of SGC-7901 cell proliferation by inhibition of telomerase activity.

Human telomerase reverse transcriptase (hTERT) is an attractive target for cancer gene therapy. However, the poor inhibition of telomerase and a time lag between the inhibition and arrest of cell proliferation limits its use in cancer gene therapy. RNA interference (RNAi) has been proposed as a potential technique for the treatment of cancer with a long-term gene silencing response induced by short hairpin RNA (shRNA). To investigate the long-term effects of telomerase inhibition through the down-regulation of the hTERT gene and the potential role of hTERT in cancer gene therapy, we constructed an shRNA-directed hTERT-expressing vector and introduced it into SGC-7901 cells. A population of cells that stably expressed the shRNA was selected by G418 and continuously cultured in a medium with half the antibiotic concentration for 3 months and the anti-proliferation effects of shRNA-targeted hTERT were then detected. The results showed that shRNA-targeted hTERT significantly inhibited cell proliferation and increased cell apoptosis by down-regulating hTERT expression, thus decreasing telomerase activity. These findings suggest that shRNA-targeted hTERT has long-term anti-proliferation effects on SGC-7901 cells, and it is a potential approach in telomerase-based gene therapy.