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BackgroundAlthough zebrafish are commonly used to study intestinal mucosal immunity, no dedicated procedure for isolating immune cells from zebrafish intestines is currently available. A speedy and simple operating approach for preparing cell suspension from mucosa has been devised to better understanding of intestinal cellular immunity in zebrafish.Methods and resultsThe mucosal villi were separated away from the muscle layer by repeated blows. The complete deprivation of mucosa was done and evidenced by HE and qPCR results. Higher expression of both innate (mpeg1, mpx, and lck) and adaptive immune genes (zap70, blnk, foxp3a, and foxp3b) was revealed compared to cells obtained by typical mesh rubbing. The cytometric results also revealed that the tested operation group had a higher concentration and viability. Further, fluorescent-labelled immune cells from 3mo Tg(lyz:DsRED2), Tg(mpeg1:EGFP), Tg(Rag2:DsRED), and Tg(lck:EGFP), were isolated and evaluated for the proportion, and immune cells’ type could be inferred from the expression of marker genes. The transcriptomic data demonstrated that the intestinal immune cell suspension made using the new technique was enriched in immune-related genes and pathways, including il17a/f, il22, cd59, and zap70, as well as pattern recognition receptor signaling and cytokine-cytokine receptor interaction. In addition, the low expression of DEG for the adherent and close junctions indicated less muscular contamination. Also, lower expression of gel-forming mucus-associated genes in the mucosal cell suspension was consistent with the current less viscous cell suspension. To apply and validate the developed manipulation, enteritis was induced by soybean meal diet, and immune cell suspensions were analyzed by flow cytometry and qPCR. The finding that in enteritis samples, there was inflammatory increase of neutrophils and macrophages, was in line with upregulated cytokines (il8 and il10) and cell markers (mpeg1 and mpx).ConclusionAs a result, the current work created a realistic technique for studying intestinal immune cells in zebrafish. The immune cells acquired may aid in further research and knowledge of intestinal illness at the cellular level.

BackgroundSignificant progress has been made in the investigation of neoadjuvant immune-chemoradiotherapy (NICRT) and neoadjuvant immune-chemotherapy (NICT) on the outcomes of esophageal cancer patients. To summarize the current developments, a systematic review and meta-analysis were conducted to evaluate the efficacy and safety of neoadjuvant immunotherapy combined with chemoradiotherapy or chemotherapy.MethodsA search strategy of prospective studies on esophageal cancer receiving neoadjuvant immunotherapy was predefined to scan PubMed, Embase, Cochrane, and additional major conferences for prospective studies. Efficacy was assessed by pathological complete response (pCR), major pathological response (MPR), and R0 resection rates. Safety was evaluated based on the incidence of grade ≥ 3 treatment-related adverse events (TRAEs), neoadjuvant therapy completion rate, surgical resection rate, and surgical delay rate. Differences between the NICRT and NICT groups were also analyzed.ResultsA total of 38 studies qualified for the analysis. The pooled pCR, MPR, and R0 resection rates were 30, 58, and 99%, respectively. The pCR and MPR in the NICRT vs. NICT group were 38% vs. 28% (p=0.078) and 67% vs. 57% (p=0.181), respectively. The pooled incidence of grade ≥ 3 TRAEs was 24% (NICRT,58%, I2 = 61% vs. NICT,18%, I2 = 79%; pConclusionsNeoadjuvant immunotherapy combined with chemoradiotherapy or chemotherapy is effective and safe in the short term for locally advanced esophageal cancer. However, further randomized trials are needed to confirm which combined model is more favorable.Systematic review registrationhttps://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42021284266, identifier CRD42021284266.

Although China possesses a large number of islands, knowledge of their overall ecological condition is limited on a national scale. Accordingly, this study developed a comprehensive and quantitative island ecosystem assessment framework that incorporates environmental quality, biological structure, and landscape patterns. The framework was applied to 42 typical, representative islands within China’s coastal regions, including different climatic zones, area sizes, populations, and substratum lithologies. Results showed that the value range of the comprehensive index of island ecosystem condition (CIisland) was significant (from 52.33 to 89.53, 71.72 average), and 85.4% of islands scored either “high” or “good,” indicative of their exceptional condition. However, conditions varied among different island components, indicating different challenges. Environmental quality index (Ienv) values ranged from 60.70 to 94.80 (79.72 average), where primary threats were seawater nutrient pollution, petroleum, and marine organism metal contamination. Biological structure index (Ibio) values ranged from 43.50 to 89.40 (66.10 average), where primary threats were vegetation destruction and a reduction in marine biodiversity within intertidal and subtidal zones. Landscape pattern index (Iland) values varied significantly (from 6 to 100, 72.36 average), where a key characteristic of highly developed islands was low natural landscape (vegetation) coverage. Conditions also varied among island climate zones, lithologies, area sizes, and levels of human habitation. Additionally, island ecosystem conditions were influenced by both natural and anthropogenic factors as well as area size, population, and GDP. Although distance to the mainland significantly correlated to (Ibio), and (CIisland), these parameters did not significantly correlate to (Ienv), which might be explained by that environmental quality was generally conditional on large-scale environments. This assessment is intended to provide a holistic perspective on island conditions in China while identifying key challenges and associated driving factors, which will greatly aid in establishing island-specific sustainable management practices.

To help prevent foodborne enteritis in aquaculture, several feed additives, such as herbal medicine, have been added to fish diets. Predictions of effective herb medicines for treating fish foodborne enteritis from key regulated DEGs (differentially expressed genes) in transcriptomic data can aid in the development of feed additives using the Traditional Chinese Medicine Integrated Database. Seabuckthorn has been assessed as a promising candidate for treating grass carp soybean-induced enteritis (SBMIE). In the present study, the SBMIE zebrafish model was used to assess seabuckthorn’s therapeutic or preventative effects. The results showed that intestinal and hepatic inflammation was reduced when seabuckthorn was added, either pathologically (improved intestinal villi morphology, less oil-drops) or growth-related (body fat deposition). Moreover, seabuckthorn may block the intestinal p53 signaling pathway, while activating the PPAR signaling pathway and fatty acid metabolism in the liver. 16S rRNA gene sequencing results also indicated a significant increase in OTU numbers and skewed overlapping with the fish meal group following the addition of seabuckthorn. Additionally, there were signs of altered gut microbiota taxa composition, particularly for reduced TM7, Sphingomonas, and Shigella, following the addition of seabuckthorn. Hindgut imaging of fluorescent immune cells in SBMIE larvae revealed the immune regulatory mechanisms at the cellular level. Seabuckthorn may significantly inhibit the inflammatory gathering of neutrophils, macrophages, and mature T cells, as well as cellular protrusions’ formation. On the other hand, in larvae, seabuckthorn inhibited the inflammatory aggregation of lck+ T cells but not immature lymphocytes, indicating that it affected intestinal adaptive immunity. Although seabuckthorn did not affect the distribution of intestinal CD4+ cells, the number of hepatic CD4+ cells were reduced in fish from the seabuckthorn supplementation group. Thus, the current data indicate that seabuckthorn may alleviate foodborne gut-liver symptoms by enhancing intestinal mucosal immunity and microbiota while simultaneously inhibiting hepatic adipose disposition, making it a potential additive for preventing fish foodborne gut-liver symptoms.

Understanding diversity patterns and associated driving factors are the critical topics in macroecology and conservation biology. Phytoplankton are highly susceptible to environmental changes in estuaries, particularly eutrophication. This study examined phytoplankton alpha and beta diversity using investigation data in May (springtime), August (summer) and November (autumn) 2009 in China’s Jiulong River estuary, where it was easily polluted because of considerable discharge from a highly dense human population and low self-purification capacity with its limited river basin area, potentially resulting in eutrophication and then influencing phytoplankton diversity. Potential influencing factors were also explored, including dissolved oxygen, salinity, nutrients, nutrient ratios, geographic and hydrologic distance, and so on. The results indicated that Shannon’s index (H’) and Pielou’s index (J) decreased from the estuary’s upper to middle and then increased from middle to lower reaches, Simpson’s (D) observed the opposite trend and species number (S) gradually increased from the estuary’s upper to lower reaches. For beta diversity, all the indices showed a gradual decrease trend from the estuary’s upper to lower reaches, where also, turnover dominated beta diversity for all seasons. It is noteworthy that the significant roles that nutrients and nutrient ratios played in shaping phytoplankton diversity patterns and the nutrient balance were characterized by excess nitrogen (N) and silicon (Si) and limited phosphorus (P), which could potentially cause diatom blooms. Findings also showed that decreasing Si concentrations can help to reduce overall pollution levels as well as the restoration of the estuary’s ecosystem better than just reducing N alone. Accordingly, this study advocates for the protection of the entire estuary system with particular emphasis on its upper reaches. Moreover, greater attention should also be paid to impacts associated with N input and nutrient ratio trade-offs to the prospective watershed management of this estuary. This study provides a practical approach to explore estuarine diversity in a comprehensive way, which can inform effective biodiversity conservation and also be applied to other marine ecosystems to better guide sustainable management and conservation practices.

Foodborne intestinal inflammation is a major health and welfare issue in aquaculture. To prevent enteritis, various additives have been incorporated into the fish diet. Considering anti-inflammatory immune regulation, an effective natural compound could potentially treat or prevent intestinal inflammation. Our previous study has revealed galantamine’s effect on soybean induced enteritis (SBMIE) and has highlighted the possible role of the cholinergic anti-inflammatory pathway in the fish gut. To further activate the intestinal cholinergic related anti-inflammatory function, α7nAchR signaling was considered. In this study, sinomenine, a typical agonist of α7nAChR in mammals, was tested to treat fish foodborne enteritis via its potential anti-inflammation effect using the zebrafish foodborne enteritis model. After sinomenine’s dietary inclusion, results suggested that there was an alleviation of intestinal inflammation at a pathological level. This outcome was demonstrated through the improved morphology of intestinal villi. At a molecular level, SN suppressed inflammatory cytokines’ expression (especially for tnf-α) and upregulated anti-inflammation-related functions (indicated by expression of il-10, il-22, and foxp3a). To systematically understand sinomenine’s intestinal effect on SBMIE, transcriptomic analysis was done on the SBMIE adult fish model. DEGs (sinomenine vs soybean meal groups) were enriched in GO terms related to the negative regulation of lymphocyte/leukocyte activation and alpha-beta T cell proliferation, as well as the regulation of lymphocyte migration. The KEGG pathways for glycolysis and insulin signaling indicated metabolic adjustments of α7nAchR mediated anti-inflammatory effect. To demonstrate the immune cells’ response, in the SBMIE larva model, inflammatory gatherings of neutrophils, macrophages, and lymphocytes caused by soybean meal could be relieved significantly with the inclusion of sinomenine. This was consistent within the sinomenine group as CD4+ or Foxp3+ lymphocytes were found with a higher proportion at the base of mucosal folds, which may suggest the Treg population. Echoing, the sinomenine group’s 16s sequencing result, there were fewer enteritis-related TM7, Sphingomonas and Shigella, but more Cetobacterium, which were related to glucose metabolism. Our findings indicate that sinomenine hydrochloride could be important in the prevention of fish foodborne enteritis at both immune and microbiota levels.