Effects of salinity acclimation on histological characteristics and miRNA expression profiles of scales in juvenile rainbow trout (Oncorhynchus mykiss).

BACKGROUND: The scales serve as an ideal model for studying the regulatory mechanism of bone homeostasis in fish. To explore the effect of salinity acclimation on bone metabolism of juvenile rainbow trout (Oncorhynchus mykiss), three sampling time points during salinity acclimation (7D, 14D and 21D) were selected to detect variations in histological characteristics. In the histological analysis, osteoblast marker enzymes alkaline phosphatase (ALP), osteoclast marker tartrate-resistant acid phosphatase (TRAcP) and calcium salt deposit areas (Von Kossa's) were detected. Changes in calcium (Ca), phosphorus (P) and the molar mass ratio of calcium to phosphorus (Ca/P) in the scales were also detected by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). In addition, the global MicroRNA (miRNA) expression profiles during salinity acclimation were examined using Illumina sequencing platform because of their important regulatory roles in teleost biological processes. RESULTS: Twelve independent miRNA libraries were constructed, a total of 664 known and 92 putative novel miRNAs were identified. A total of 290 differentially expressed (DE) miRNAs were found in clusters with significant trends in the cluster analysis, and five types of clustering patterns were obtained; 22,374 DE predicted target genes of the aforementioned 290 DE miRNAs were obtained, 5957 of which clustered in six types of clustering patterns with a significant trend. To better understand the functions of the DE miRNAs, GO and KEGG analysis was performed on the 5957 target genes, as a result, they were significantly enriched in bone metabolism related signaling pathways such as MAPK signaling pathway, Calcium signaling pathway, Wnt signaling pathway, Mineral absorption and NF-kappa B signaling pathway. Six DE miRNAs were randomly selected and their expression were verified by quantitative real-time PCR (qRT-PCR), the expression trends were consistent with the results of transcriptome sequencing. CONCLUSIONS: The DE miRNAs and DE target genes identified in this study might play an important role in regulation of bone metabolism during salinity acclimation, relative genes or pathways could serve as key candidates for further studies to elucidate molecular mechanism of teleost bone metabolism, and help performing salinity acclimation and developing marine culture of salmonid species.

Intestinal microbiota pathogenesis and fecal microbiota transplantation for inflammatory bowel disease.

The intestinal microbiota plays an important role in inflammatory bowel disease (IBD). The pathogenesis of IBD involves inappropriate ongoing activation of the mucosal immune system driven by abnormal intestinal microbiota in genetically predisposed individuals. However, there are still no definitive microbial pathogens linked to the onset of IBD. The composition and function of the intestinal microbiota and their metabolites are indeed disturbed in IBD patients. The special alterations of gut microbiota associated with IBD remain to be evaluated. The microbial interactions and host-microbe immune interactions are still not clarified. Limitations of present probiotic products in IBD are mainly due to modest clinical efficacy, few available strains and no standardized administration. Fecal microbiota transplantation (FMT) may restore intestinal microbial homeostasis, and preliminary data have shown the clinical efficacy of FMT on refractory IBD or IBD combined with Clostridium difficile infection. Additionally, synthetic microbiota transplantation with the defined composition of fecal microbiota is also a promising therapeutic approach for IBD. However, FMT-related barriers, including the mechanism of restoring gut microbiota, standardized donor screening, fecal material preparation and administration, and long-term safety should be resolved. The role of intestinal microbiota and FMT in IBD should be further investigated by metagenomic and metatranscriptomic analyses combined with germ-free/human flora-associated animals and chemostat gut models.

[The effects of early enteral nutrition with addition of probiotics on the prognosis of patients suffering from severe acute pancreatitis].

OBJECTIVE: To investigate the curative effect of early enteral nutrition (EN) supplemented with probiotics (bifidobacterium) in patients with severe acute pancreatitis (ASP). METHODS: Seventy SAP cases admitted from January 2005 to October 2012 were randomly assigned into parenteral nutrition (PN) group (n=22), EN group (n=25) and bifidobacterium added EN (P+EN) group (n=23). In P+EN group, patients were given their nutrition the same as that of EN, and also probiotics (bifidobacterium, 4 capsules every 12 hours, given through nasal gastric tube, each capsule weighing 210 mg). The routine treatment including anti-infection and anti-acid agents, and that of inhibition of pancreatic secretion were given, except for the different nutritional interventions in all groups. The blood samples were collected for e same measurements of interleukin-8 (IL-8) and tumor necrosis factor (TNF-α) by enzyme linked immunosorbent assay (ELISA), and for the C-reactive protein (CRP), lactic acid dehydrogenase (LDH), white blood cell (WBC) count, amylase and lipase by biochemistry assay 1 day before intervention of nutrition, and 7 days and 14 days after intervention. Changes in organ function and outcome were also recorded at the same time points. RESULTS: The plasma levels of IL-8, TNF-α, CRP, LDH, WBC count, amylase and lipase were significantly reduced after nutritional intervention compared with their levels on day 1 before intervention in all three groups. The plasma IL-8, TNF-α, CRP, lipase, LDH at 14 days after intervention of nutrition in P+EN group were significantly lower than those in PN group and EN group (IL-8: 21.00 ± 7.07 µg/L vs. 48.00 ± 10.32 µg/L, 32.00 ± 9.30 µg/L; TNF-α: 44.3 ± 10.9 ng/L vs. 132.1 ± 34.1 ng/L, 67.8 ± 22.3 ng/L; CRP: 35.0 ± 12.4 mg/L vs. 103.2 ± 49.2 mg/L, 63.0 ± 29.2 mg/L; lipase: 269 ± 79 U/L vs. 670 ± 145 U/L, 310 ± 78 U/L; LDH: 21.8 ± 10.3 U/L vs. 78.1 ± 37.4 U/L, 37.9 ± 25.1 U/L, P<0.05 or P<0.01). The WBC count in P+EN group was significantly lower than that in PN group (5.9 ± 3.0 × 109/L, 6.3 ± 3.2 × 109/L vs. 9.6 ± 3.0 ×109/L, both P<0.05), but there was no significant difference in amylase between P+EN group and PN group (211 ± 49 U/L, 236 ± 52 U/L vs. 298 ± 71 U/L, P>0.05). The gastrointestinal dysfunction score in P+EN, EN, PN groups 14 days after nutritional intervention was 0.28 ± 0.05, 0.43 ± 0.09, 0.71 ± 0.11, respectively, with statistically significant differences (all P<0.01). Compared with PN and EN groups, the incidence of upper gastrointestinal bleeding (1 vs. 9, 2), infection and abscess (2 vs. 12, 5) was lower (all P<0.01), and hospital day was significantly shortened in P+EN group (10.4 ± 3.9 days vs. 25.8 ± 6.4 days, 13.4 ± 5.2 days, both P<0.01). There was no significant statistical difference in mortality rate among three groups. CONCLUSION: Our results indicated that early EN with addition of probiotics (bifidobacterium) resulted in significant lowering of the level of pro-inflammatory cytokines, earlier restoration of gastrointestinal function, decrease of complications such as infection, and shortening of hospital day in patients with SAP.

[Evaluation of the effect of ilaprazole on intragastric pH in patients with duodenal ulcer].

OBJECTIVE: To evaluate the effect of ilaprazole enteric tablets on intragastric pH in duodenal ulcer patients. METHODS: A randomized, double blind, positive controlled clinical trial was carried out. A total of forty-two patients with duodenal ulcer were randomized into low dose ilaprazole group (5 mg/d), medium dose ilaprazole group (10 mg/d), high dose ilaprazole group (20 mg/d) and omeprazole group (20 mg/d). An ambulatory 24 hour intragastric pH study was performed at the fifth treatment day. Fraction time pH above 3, 4 or 5, median values of 24 hour diurnal pH and 12 hour nocturnal pH, the percentage of patients with total time pH above 3, 4 or 5 at least for 18 hours were evaluated. RESULTS: There were no significant differences of fraction time pH above 3 or 4, median values of 24 hour diurnal pH and 12 hour nocturnal pH and the percentage of patients with total time pH above 3, 4 or 5 at least for 18 hours among all the groups with different doses of ilaprazole and the omeprazole group. The fraction time pH above 5 in medium and high dose ilaprazole groups were (87.96 + or - 12.29)% and (89.86 + or - 15.18)% respectively, which was higher than that in low dose ilaprazole group [(67.17 + or - 30.16)%] and omeprazole group [(76.14 + or - 16.75)%], P < 0.05. CONCLUSION: Ilaprazole has a strong effect on intragastric acid control with a dose dependent trend.