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The dipeptide dl-methionyl-dl-methionine (Met-Met) has extremely low water solubility and better absorption than other methionine sources (such as dl-methionine and l-methionine) available in the market. Therefore, six diets (D1, D2, D3, D4, D5 and D6) containing 0, 0·07, 0·15, 0·21, 0·28 and 0·38 % Met-Met were formulated to investigate the effects of Met-Met in juvenile Nile tilapia, Oreochromis niloticus (17 g initial body weight). The results indicated that percentage weight gain and specific growth rate of fish fed with D2 and D3 diets were higher than those fed with D1, D4–D6 diets. The levels of total essential amino acid in whole body of fish fed with D3 and D4 diets were significantly higher than those fed the D1 diet. Superoxide dismutase activity and malondialdehyde content have no significant difference in fish fed the diet with or without Met-Met supplementation. Majority of reads derived from the fish intestine belonged to members of Fusobacteria, followed by Bacteroidetes and Proteobacteria. Diversity of intestinal microbiota and total antioxidant capacity in fish fed with D3 diet was significantly higher than that of other groups. Based on the growth results, the authors conclude that the optimal level of Met is 0·61 % Met with the addition of 0·15 % Met-Met for grower-phase O. niloticus.

A novel and efficient redox reaction was developed to react 5-amino-1H-pyrazole-4-carbaldehyde with sodium nitrite (NaNO2) in an acidic solution (HCl/MeOH) to generate 5-amino-4-nitrosopyrazole, pyrazole-4-carbaldehyde, or diazenylpyrazole selectively. The results showed that 5-amino-4-nitrosopyrazoles were formed as the major product in the diluted acidic solution (≤2 N HCl in MeOH solution) through redox, formylation, and nitrosation reactions of NaNO2. Intriguingly, pyrazole-4-carbaldehyde was the main product under 6 N HCl in MeOH solution.

Nrf2 is an important component of Nrf2-ARE signaling pathway. However, information about Nrf2 in golden pompano (Trachinotus ovatus) is quiet limited. Thus, the full length of Nrf2 was cloned from T. ovatus in this study. The full-length cDNA sequence was 2376 bp in length and comprised of a 1848 bp ORF preceded by 123 bp 5′UTR, and followed by a 405 bp UTR. The putative amino acid sequence of the Nrf2 contained 615 residues, consisting of a conservative basic region leucin zipper. The N terminal portion of Nrf2 is rich in glutamic and aspartic acids and functioned as an acidic type of activation domain. The C terminus of the protein contained a region with heptad repeats of leucines. The phylogenetic tree showed that Seriola dumerili and T. ovatus were clustered as a branch, and combined with Mastacembelus armatus, Oreochromis niloticus and Takifugu rubripes. This branch of 5 species clustered with other branch of Oncorhynchus mykiss and Salmo salar. Nrf2 gene was predominantly expressed in liver and gut. The relative expression of Nrf2 in liver was significantly higher than that in brain, kidney, spleen and heart. After air-exposure, the expression level of HIF-1 alpha, keap1, Nrf2, HO-1, SOD and GSH-PX were significantly higher in air-exposure group than that in control group. This present study showed that Nrf2 plays a pivotal role in oxidative stress caused by air-exposure in T. ovatus, providing information about response to oxidative stress on the basis of the Nrf2-ARE pathway in T. ovatus.

The present study investigated the potential mechanisms of astaxanthin in the regulation of gastrointestinal immunity and retinal mitochondrial function of golden pompano (Trachinotus ovatus). Triplicate groups of juvenile T. ovatus (mean initial weight: 6.03 ± 0.01 g) were fed one of six diets (D1, D2, D3, D4, D5, and D6) for 8 weeks, with each diet containing various concentrations of astaxanthin (0, 0.0005, 0.001, 0.005, 0.01, or 0.1%, respectively). Growth performance of fish fed the D2–D5 diets was higher than that of fish fed the D1 diet; however, growth performance and survival of fish deteriorated sharply in fish fed the D6 diet. Gut villus in fish fed the D2–D5 diets were significantly longer and wider than that of fish fed the D6 diet. Feeding with D2–D5 diets led to increased abundance of Bacillus, Pseudomonas, Oceanobacillus, Lactococcus, Halomonas, Lactobacillus, and Psychrobacter while abundance of Vibrio and Bacterium decreased. Additionally, feeding with the D6 diet resulted in a sharp decline in Pseudomonas and Lactobacillus abundance and a sharp increase in Vibrio abundance. A low dissolved oxygen environment (DO, 1.08 mg/L) was conducted for 10 h after the rearing trial. No fish mortality was observed for any of the diet treatments. Lysozyme (LZY) activity in fish fed the D6 diet decreased sharply and was significantly lower than that in other groups. ROS production also decreased sharply in fish fed the D6 diet. Moreover, the conjunctiva and sclera in the fish fed the D6 diet were indistinguishable. Suitable dietary astaxanthin supplementation levels (0.005–0.1%) exerting a neuroprotective effect from low dissolved oxygen environments is due to up-regulated expression of anti-apoptotic factors, such as phosphorylated Bcl-2-associated death promoter (pBAD), phosphorylated glycogen synthase kinase-3β (pGSK-3β), Bcl-2 extra large (Bcl-xL), and down-regulated expression of Bcl-2-associated X protein (Bax) pro-apoptotic factor in retinas. Furthermore, suitable dietary astaxanthin levels (0.0005–0.01%) suppressed up-regulation of critical mitochondrial components, such as peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α), mitochondrial transcription factor A (TFAM), and mitochondrial DNA (mtDNA), while excessive astaxanthin supplementation produces the opposite effect. In brief, high-dose astaxanthin arouses and aggravates low dissolved oxygen-induced inflammation, oxidative stress, intestinal disorder, retinal apoptosis, and retinal mitochondrial dysfunction in T. ovatus. Second-degree polynomial regression of WG indicated that the optimum dietary astaxanthin for juvenile T. ovatus is 0.049%.

A nutritional feeding experiment was conducted to determine growth performance and antioxidant capacity in T. ovatus. Five diets were formulated (D1, control; D2, Saccharina japonica; D3 Undaria pinnatifida; D4 Gracilaria lemaneiformis; D5, Porphyra haitanensis) to feed fish for 8 weeks. Results showed that final body weight and weight gain in D3 and D5 groups were significantly higher than that in D1 and D4 groups (P 0.05). Feed conversion ratio in D5 group was significantly lower than that in D1 group (P 0.05). Feed intake and survival rate showed no significantly difference among all groups (P

The efficient acid catalytic synthesis of pyrazolo [3,4-d]pyrimidine was developed by treating 1H-pyrazol-5-yl-N,N-dimethylformamidine with various aminating agents including N,O-bis(trimethylsilyl)hydroxylamine (NHSiMe3(OSiMe3)), cyanamide (NH2C N), hydroxylamine (NH2OH), methoxyamine (NH2OMe), hydrazine (NH2NH2), and urea (NH2C(O)NH2) in acidic solution at reflux. Based on the experimental result, cyanamide (NH2C N) and methanesulfonic acid were indicated the best aminating agent and acid mediated solvent. On the other hands, the reactivity tendency was involved the activity of original leaving species grafting on the aminating agents, such as –OH, –OMe, –OSiMe3, –NH2, –OSiMe3, –C(O)NH2, and –C N, in acid catalytic heterocyclic reaction.

Two trials were conducted to determine the effects of dietary Forsythia suspensa extract (FSE) on shrimp, Penaeus monodon, first on growth performance, second on the immune response and immune related gene expression of shrimp. In trial 1, shrimp (mean initial wet weight about 3.02 g) were fed with five diets containing 0% (basal diet), 0.01%, 0.02%, 0.04% and 0.06% FSE in triplicate for 60 days. Growth performance (final body wet weight, FBW; weight gain, WG; biomass gain, BG) of shrimp fed FSE diets were higher (P 0.05). The antioxidant capacity (total antioxidant status, TAS; glutathione peroxidase, GSH-Px) appears in the trend of firstly increasing then decreasing with the increasing of dietary FSE levels. The highest value of TAS and GSH-Px were found in shrimp fed 0.02% FSE diet and were significantly higher than that of shrimp fed the basal and 0.06% FSE diets (P 0.05). In trial 2, a pathogenic strain of Vibrio parahaemolyticus 3HP (VP3HP) injection challenge test was conducted for 6-day after the rearing trial and shrimp survival were also compared among treatments. Survival of shrimp fed diets supplemented with 0.01%–0.02% FSE were higher than that of shrimp fed the basal and 0.06% FSE diets (P 0.05). The molecular gene expression profile of Hsp 70 mRNA was increased while HIF-1α mRNA was decreased when compared to trial 1. In conclusion, results suggested that dietary intake containing FSE could enhance the growth performance and antioxidant capacity of P. monodon and furthermore reduce oxidative stress and immune depression challenged by a pathogenic strain of Vibrio parahaemolyticus stress. Considering the effect of FSE on both growth performance and immune response of P. monodon, the level of FSE supplemented in the diet should be between 0.01% and 0.02%.

Purpose To evaluate the utility of the whole-lesion histogram apparent diffusion coefficient (ADC) for characterizing the heterogeneity of mucinous breast carcinoma (MBC) and to determine which ADC metrics may help to best differentiate subtypes of MBC. Materials and Methods This retrospective study involved 52 MBC patients, including 37 pure MBC (PMBC) and 15 mixed MBC (MMBC). The PMBC patients were subtyped into PMBC-A (20 cases) and PMBC-B (17 cases) groups. All patients underwent preoperative diffusion-weighted imaging (DWI) at 1.5T and the whole-lesion ADC assessments were generated. Histogram-derived ADC parameters were compared between PMBC vs. MMBC and PMBC-A vs. PMBC-B, and receiver operating characteristic (ROC) curve analysis was used to determine optimal histogram parameters for differentiating these groups. Results The PMBC group exhibited significantly higher ADC values for the mean (P = 0.004), 25th (P = 0.004), 50th (P = 0.004), 75th (P = 0.006), and 90th percentiles (P = 0.013) and skewness (P = 0.021) than did the MMBC group. The 25th percentile of ADC values achieved the highest area under the curve (AUC) (0.792), with a cutoff value of 1.345 × 10−3mm2/s, in distinguishing PMBC and MMBC. The PMBC-A group showed significantly higher ADC values for the mean (P = 0.049), 25th (P = 0.015), and 50th (P = 0.026) percentiles and skewness (P = 0.004) than did the PMBC-B group. The 25th percentile of the ADC cutoff value (1.476 × 10−3mm2/s) demonstrated the best AUC (0.837) among the ADC values for distinguishing PMBC-A and PMBC-B. Conclusion Whole-lesion ADC histogram analysis enables comprehensive evaluation of an MBC in its entirety and differentiating subtypes of MBC. Thus, it may be a helpful and supportive tool for conventional MRI. Level of Evidence: 4 Technical Efficacy Stage 2 J. Magn. Reson. Imaging 2017.

The aim of the present study was to investigate the differences of the pathological changes and cognitive function after bilateral common carotid artery occlusion (BCCAO) between Sprague-Dawley (SD) and Wistar rats. Male SD and Wistar rats were randomly divided into 2 groups, respectively: sham operated (S-sham and W-sham) and operated (S-BCCAO and W-BCCAO) groups. The survival rate and the rate of loss of pupillary light reflex (PLR) were observed on day 1, 3, 7, 14 and 28 after the operation, and the light-dark box, Y-maze and odor recognition tests were performed to detect cognitive function on day 28 after the operation. HE and Luxol fast blue staining were used to observe the pathological changes of gray matter (hippocampus), white matter (optical tract), optic nerve, and retina. The results showed that the survival rate of the W-BCCAO group was 62.5%, and PLR loss rate was 100%; whereas the survival rate of the S-BCCAO group was 100%, and PLR loss rate was 58.3%. In the W-BCCAO group, percentages of time spent and distance traveled in the light box were more than those in the W-sham group, but there was no statistical significance between the S-BCCAO and S-sham groups. In the S-BCCAO group, the percentages of time spent and distance traveled in the III arm (labyrinth arm) of the Y-maze were less than those in the S-sham group, but no statistical significance was found between the W-BCCAO group and W-sham group. In the S-BCCAO group, the discrimination ratio of the odor recognition task was less than that in the S-sham group, but no statistical significance could be seen between the W-BCCAO and W-sham groups. Ischemic injury was observed in the CA1 area of the hippocampus in the S-BCCAO group, but no readily visible damage was observed in the W-BCCAO group. Ischemic injury of the visual beam and optic nerve was observed in both the S-BCCAO and W-BCCAO groups. Compared with the corresponding sham groups, the S-BCCAO and W-BCCAO groups showed serious retinal damage with significant thinner retina. The ganglion cell layer (GCL), inner plexiform layer (IPL), and outer plexiform layer (OPL) were thinner in the S-BCCAO group, but no statistical significances were shown in the other layers. All the layers, except the outer nuclear layer (ONL), were significantly thinner in the W-BCCAO group. The results indicate that there are differences of the pathological changes in the hippocampus and visual conduction pathway after BCCAO between SD and Wistar rats, and the degree of learning and memory injury was also different, which suggests that the vascular dementia model of different rat strains should be selected according to research purpose.

The detection of drug-induced anaphylactoid reactions remains a global challenge,still lacking mature and reliable animal models or test methods. Therefore,the purpose of this paper is to explore and establish the test methods and evaluation standards for anaphylactoid reactions that apply to injection drugs. Based on the anaphylactoid reaction symptoms of mice induced by intravenous injection drugs C48/40 and Tween 80,a list of systemic anaphylactoid reaction symptoms in mice was sorted out and an evaluation standard of anaphylactoid reactions symptoms was established by applying symptom intensity coefficient K( that can represent these verity of anaphylactoid reaction symptoms) and its calculation formula Accordingly,histamine,tryptase,and Ig E were selected as blood indicators of anaphylactoid reactions,so that a test method combining symptoms evaluation and blood makers detection was established.This test method could be used to evaluate the characteristics of anaphylactoid reactions: coefficient K,blood histamine levels were highly and positively correlated with C48/80 and Tween 80 dose; The log value of histamine was highly and positively correlated with K; tryptase level may rise,or remain steady,or drop,possibly associated with the characteristics of the tested object and time for blood taking; and Ig E level would drop or remain steady,but it would not rise,which can be clearly distinguished from type I allergic reactions. On this basis,tiohexol,iopromide,paclitaxel,Xuesaitong Injection,Shuanghuanglian Injection and Shengmai Injection were used to investigate the applicability. The testing results showed a high degree of consistency with the actual clinical situation. The results suggest that the method of systemic anaphylaxis test in mice has high sensitivity,specificity and good consistency with clinical practice.It is suggested to be further validated and popularized.

A feeding experiment was conducted to determine the effects of Schizochytrium sp. on growth performance, antioxidant capacity and non-specific immunity in golden pompano (Trachinotus ovatus).Two diets were formulated with or without Schizochytrium sp. supplemented (D1:0% and D2: 3%) to feed fish for 8 weeks. Results showed that growth performance, feed intake and survival rate increased significantly with Schizochytrium sp. supplemented (P 0.05). Feed coefficient rate (FCR) of golden pompano fed the diet supplemented with Schizochytrium sp. was significantly lower than that of fish fed the control diet (P 0.05). No significant differences were found in antioxidant capacity both in transcriptional level, including nclear factor erythroid-2-related factor-2 (Nrf2), Kelch-like-ECH-associated protein (keap1), catalase (CAT), glutathione peroxidase (GSH-PX) and heme oxygenase 1 (HO-1) and enzyme activity, such as total antioxidant capacity (T-AOC), malondialdehyde (MDA) and superoxide dismutase (SOD) (P 0.05). Gut amylase and lipase were significantly higher in dietary Schizochytrium sp. supplemented treatment than that in control group (P 0.05). The relative peroxisome proliferator-activated receptor-α (PPARα) expression level in liver was significantly higher in Schizochytrium sp supplemented treatment than that in control one (P 0.05). The mRNA expression of myeloid differentiation factor 88 (MyD88), IL-1R-associated kinases 4 (IRAK4), interferon regulating Factor 3 (IRF3), interferon regulating Factor 3(IRF7) and heat shock protein 70 (HSP70) were significantly lower in Schizochytrium sp. supplemented treatment than that in control one (P 0.05). In Schizochytrium sp. supplemented diet, golden pompano had significantly longer villi length than that in control diet (P 0.05); muscle thickness in Schizochytrium sp. supplemented diet was thicker than that in control one (P 0.05) and there were more goblet cells in Schizochytrium sp. treatment (P 0.05). After the rearing trial, an air exposure trial was conducted. Results showed that the air-exposure mortality (AEM) and mRNA expression level of Nrf2, keap1, CAT, GSH-PX and HO-1 showed no significant difference (P 0.05). These results indicated that dietary Schizochytrium sp. improved the growth performance and non-specific immunity of golden pompano while made no difference to antioxidant capacity.

Heme oxygenase-1(HO-1) is a stress-inducible enzyme that mediates antioxidative and cytoprotective effects to maintain cellular redox homeostasis. In the present study, the full sequence of HO-1 was cloned from golden pompano(Trachinotus ovatus) by RT-PCR and RACE-PCR. The full cDNA sequence of HO-1 was 1349 bp in length which comprised of a 726 bp open reading frame (ORF) preceded by 262 bp 5′-untranslated region (UTR), and followed by a 360 bp 3′UTR, encoding 241 amino acid residues. Phylogenetic analysis revealed that HO-1 showed highest similarity to that of Takifugu rubripes. Tissue distribution analysis showed that the expression level of HO-1 was relatively high in heart, liver and spleen. A trial was conducted to investigate the response of Nrf2/HO-1 signaling pathway to oxidative stress induced by copper. The results showed that mRNA expression of NF-E2-related nuclear factor2 (Nrf2), Kelch-like-ECH-associated protein1 (keap1), superoxide dismutase (SOD), catalase (CAT), HO-1, NAD(P)H quinone oxidoreductase 1 (NQO1) and Glutathione peroxidase (GSH-PX) all significantly increased in copper treated group than that in the control group. This work provides new insight into the molecular mechanism underlying the Nrf2/HO-1 pathway in oxidative response in T. ovatus.

This study was conducted to elucidate the effects of dietary mixed probiotics on growth, non-specific immunity, intestinal morphology and microbiota of juvenile pacific white shrimp, Litopenaeus vannamei. Juvenile shrimp (initial body weight 1.21 ± 0.01 g) were fed diets containing graded probiotics (F1: 0 mg/kg probiotics; F2: 1000 mg/kg probiotics; F3: 2000 mg/kg probiotics; F4: 4000 mg/kg compound probiotics; F5: 6000 mg/kg probiotics; F6: 8000 mg/kg probiotics) for 8 weeks. The result of this trial showed that the growth performance (SGR, WG, FBW) of shrimp fed diets containing probiotics (F2∼F6) were significantly higher than that of shrimp fed diet without supplemental probiotics (F1) (P 0.05), and the highest values of the growth performance (SGR, WG, FBW) and lowest FCR were found in shrimp fed the diet containing 2000 mg/kg probiotics. Total antioxidant capacity of shrimp fed diet F2 and F3 were significantly higher than that of shrimp fed the basal diets (P 0.05). Superoxide dismutase in F4 treatment was significantly higher than that of basal treatment (P 0.05). Catalase of shrimp in all probiotics supplemented (F2∼F6) treatments were significantly higher than that of the control one (F1) (P 0.05). Malondialdehyde in F5 groups was significantly lower than that of F1 groups (P 0.05). Alkline phosphatase and acid phosphatase in F3 treatments were significantly higher than those of the basal one (P 0.05). Lysozyme of shrimp fed F2∼F6 were significantly higher than that of shrimp fed F1 diet (P 0.05). The lipase and amylase activities in 2000 mg/kg probiotics groups showed the highest activities and were significantly higher than that of control one (P 0.05). Intestinal villi height in F3∼F6 treatments were significantly higher than that of control one (P 0.05). Alpha diversity indices including observed species, chao1, ACE and shannon indices showed that F2 and F3 groups had higher microbial diversity in their intestines, both richness and evenness. PCA plot showed that there was a clear shift of F2 and F3 groups from the control groups in microbial community structure. The dominant phyla in pacific white shrimp are proteobacteria, bacteroidetes and actinobacteria, the dominant genus were algoriphagus and vibrio. As the probiotics increased, the gemmatimonadetes, acidobacteria, deltaproteobacteria and xanthomonadales firstly increased and then decreased, with the highest content in F2 group, which was no significant difference to F3 group (P 0.05) while significantly higher than other groups (P 0.05). In conclusion, the supplement of mixed species probiotics can promote growth performance, enhance the non-specific immunity, influence the microbiota of the pacific white shrimps and the recommended optimum dosage in diet of Litopenaeus vannamei was 2000 mg/kg.

An 8 weeks feeding experiment was conducted to evaluate the effects of dietary supplementation with hydrolyzed yeast (HY) (Rhodotorula mucilaginosa) on growth performance, hematological parameters, immune response and antioxidant ability of juvenile Nile tilapia. Five isonitrogenous and isolipidic diets (32% protein and 4% lipid) with different levels (0%, 0.125%, 0.25%, 0.5%, 1%) of HY were formulated. Each diet was randomly assigned to quadruplicate groups of fish (initial body weight 19.1 ± 0.01 g). Results indicated that significantly higher specific growth rate (SGR) and lower feed conversion rate (FCR) were obtained in fish fed 1% HY diet than that of fish fed 0% HY diet (P 0.05). Fish fed 0.25% HY diet showed the lowest value of hepatopancreas somatic indices (HSI) and significantly lower than that of fish fed 0% HY diet (P 0.05). Meanwhile, protein and ash in the whole-body content of fish fed 1% HY diet was significantly higher than that of fish fed 0%-0.5% HY diets. Serum immunological parameters showed that the lysozyme (LZM) activity and Complement C3 content were significantly increased by dietary supplementation of 0.5%-1% HY (P 0.05). However, dietary supplementation with 0.125%-1% HY significantly decreased the activity of myeloperoxidase (MPO) (P 0.05). Antioxidant status in serum and liver was significantly enhanced by dietary supplementation of 0.25%-1% HY through the remarkably improved superoxide dismutase (SOD) activity both in serum and liver, the raised total antioxidative capacity (T-AOC) of serum as well as the notably reduced malondialdehyde (MDA) content in the liver (P 0.05). However, T-AOC in the liver was not significantly influenced among all diet treatments (P 0.05). Villi height and intraepithelial lymphocytes (IEFs) of mid-intestine were significantly higher in fish fed 0.5%-1% HY diets (P 0.05). The challenge test demonstrated the enhanced protection against Streptococcus iniae strain by the obtained higher cumulative survival rate. In conclusion, dietary supplementation of 1% HY could maintain the better growth performance, nutrient composition as well as immune response and antioxidant capacity for juvenile Nile tilapia.

The study was conducted to compare and evaluate effects of four different macro-algaes on growth, immune response, and intestinal microbiota of Litopenaeus vannamei. In the rearing trial 1, shrimp were fed five diets containing four sources of macro-algaes for 8 weeks, named D1 (without macro-algae), D2 (Porphyra haitanensis), D3 (Undaria pinnatifida), D4 (Saccharina japonica), and D5 (Gracilaria lemaneiformis), respectively. Growth performance of shrimp in D5 diet was significantly higher than that of shrimp fed the control and D4 diet (P < 0.05); however, there is no significant difference among D2, D3, and D5 diets (P > 0.05). Apparent digestibility coefficients of dry matter from the D2, D3, and D5 diets were significantly higher than that from the control and D4 diets (P < 0.05). Supplementary macro-algaes enhanced hepatopancreas immunity through positively increasing total antioxidant status (TAS) and prophenoloxidase activity (ProPO), as well as up-regulating the hepatopancreas RNA expression of ProPO and IκBα and down-regulating the expression of transforming growth factor β. Furthermore, dietary macro-algaes modified intestinal microbiota of L. vannamei, boosting the relative abundance of beneficial bacterial such as Bacteroidetes, Firmicutes, and Bacillaceae, and decreasing those detrimental bacterial such as Gammaproteobacteria and Vibrionaceae. In the white spot syndrome virus (WSSV) challenge trial, shrimps were injected for 6-day after the rearing trial. On the fourth day, shrimp death started to occur, and the mortality in D2, D3, and D5 diets was significantly lower than that in control and SJ diets during 4–6 challenged days (P < 0.05). Dietary macro-algaes ameliorated hepatopancreas damage in L. vannamei by increasing TAS and ProPO activities and decreasing SOD activity, inhibiting the lipid peroxidation (malondialdehyde), as well as regulating the immune-related genes expression. Taken together, dietary macro-algaes availably relieved enterohepatic oxidative damage by improving antioxidant ability and immunity and regulated intestinal microbiota in L. vannamei. These results indicated that G. lemaneiformis is the most suitable macro-algae and then followed by U. pinnatifida and P. haitanensis as the feed ingredient for L. vannamei.

The present study was conducted to evaluate the effects of dietary lipid levels on lipid metabolism, growth performance, antioxidant ability and intestinal morphology of rainbow trout (Oncorhynchus mykiss) under cage culture with flowing water. O. mykiss (average initial body weight 17.64 ± 0.05 g) were fed with six diets (L1, L2, L3, L4, L5 and L6) containing six levels of lipid (10.03 %, 12.97 %, 17.22 %, 20.16 %, 23.19 %, 26.06 %) for 8 weeks. Result showed that weight gain (WG), specific growth rate (SGR) and feed intake (FI) of fish were increased with increasing of dietary lipid levels (P < 0.05). Fish fed D6 diet had higher viscerosomatic index (VSI) and mesenteric fat index (MFI) compared with fish fed other diets. The moisture and crude protein contents in whole body and muscle decreased with increasing of dietary lipid levels, but crude lipid content showed the opposite trend. The hepatic superoxide dismutase (SOD) activity was increased with increasing of dietary lipid levels (P < 0.05), while no significant differences were found in hepatic malondialdehyde (MDA) content (P> 0.05). Correspondingly, the mRNA relative expression of SOD in liver was increased with increasing of dietary lipid levels (P < 0.05). Fish fed the diet (L6) containing 26.06 % lipid level showed the optimal intestinal morphology not only in villus amount and height but also in muscular layer thickness. The lipid metabolism related genes expression such as fatty acid synthetase (FAS), lipoprotein lipase (LPL), were significantly affected by dietary lipid levels (P < 0.05). The mRNA relative expression of FAS was increased from L1 to L4 and then decreased with the increasing of dietary lipid levels. The mRNA expression of LPL in fish fed with L5 and L6 diets were significantly higher than other treatments. These results indicated that the suitable lipid requirement of rainbow trout under cage culture with flowing water was not less than 26.06 %.

Carbohydrates are the cheapest source of dietary energy for fish. Utilization of carbohydrates in feed is beneficial to sparing protein consumption as energy source and saving feed cost. Therefore, 8-week feeding experiment was conducted to investigate the effects of starch level on growth performance, antioxidant capacity, serum biochemical parameters, intestinal morphology and intestinal microbiota of Trachinotus ovatus. Six isonitrogenous and isolipidic diets (D1, D2, D3, D4, D5 and D6) containing 0%, 5%, 10%, 15%, 20% and 25% corn starch were formulated, respectively. Each diet was randomly assigned to triplicate groups of fish (initial body weight 11.84 g) fed twice daily to apparent satiation. Results showed that percent weight gain, specific growth ratio, hepatosomatic index, total antioxidation capacity and glutathione content of fish fed with D6 diet were the highest among all experimental treatments. Feed conversion rate was significantly lower in fish fed with D5 and D6 diets, than in fish fed with D1-D4 diets. No significant differences were found in the survival rate, viscerasomatic index, malondialdehyde and serum triglyceride contents among all diet treatments. The superoxide dismutase activity of T. ovatus fed with D3-D6 diets were relatively higher than those fed with D1 and D2 diets. Fish fed with D3 diet showed the highest value of serum total cholesterol. High-density lipoprotein cholesterol / low-density lipoprotein cholesterol ratio of fish fed with D1 and D6 diets were relatively higher than those fed with D2-D5 diets. No significant pathological changes in the foregut were observed among all diet treatments. Dietary starch level lead to alterations in the intestinal microbiota in T. ovatus. D2-D5 dietary treatments increased the abundance of harmful bacteria such as Vibrio, Photobacterium, Mycoplasma. D1 and D6 dietary treatments had similar intestinal microbiota composition. In conclusion, 25% starch level was beneficial to the growth and health of T. ovatus.

The present study was conducted to evaluate a multi-strain probiotic (MP) on growth performance, immune and antioxidant function, response to hypoxia stress and resistance to Aeromonas hydrophila of grass carp (Ctenopharyngodon idella). Based on the viable cell counts of aerobic Bacillus spp., six experimental diets with MP supplemented at 0, 0.34, 1.68, 3.36, 6.72, 10.1 g kg−1 were formulated and 900 juveniles (7.30 ± 0.01 g) were equally distributed into 30 aquaria with respective diet for 60 days. Results showed that fish with 0.34–1.68 g kg−1 MP had better growth and feed utilization. Further, plasma total protein, albumin and high-density lipoprotein were remarkably increased with dietary MP at >1.68 g kg−1. Dietary MP supplementation at 6.72–10.1 g kg−1 strikingly elevated plasma myeloperoxidase activity and complement C3 content. For fish with MP at 1.68 and 6.72–10.1 g kg−1, their liver malondialdehyde and glutathione peroxidase were remarkably declined and promoted. After hypoxia stress, fish with 3.36–6.72 g kg−1 MP showed significantly higher respiratory burst activity. Challenge test by A. hydrophila confirmed the protection effects of MP through the decreased cumulative mortality rates. For intestinal histomorphology and enzymatic analyses, fish with 1.68 g kg−1 MP displayed significantly higher intestinal villi height, goblet cells and alkaline phosphatase activity. In conclusion, dietary MP supplementation at 1.68 g kg−1 could promote growth, intestinal morphology and antioxidant capacity, while enhancing host immunity requires higher dosages of MP. Broken-line analysis of weight gain revealed that 1.34 g kg−1 is the optimum dosage for the growth of grass carp.

p65 is an important component of NF-κB signaling pathways. However, information about p65 subunit in golden pompano (Trachinotus ovatus) is quiet limit. Thus, the full length of p65 subunit was cloned from T. ovatus in this study. The full-length cDNA sequence was 3162 bp in length and comprised of a 1886 bp open reading frame (ORF) preceded by 153 bp 5′-untranslated region (UTR), and followed by a 1143 bp 3′UTR. The p65 of T. ovatus contains a conserved N-terminal REL-homology domain (RHD) with 273 amino acids. The phylogenetic tree showed that p65 in T. ovatus was clustered in the clade of that in marine fish. mRNA expression of p65 was detected in 12 different tissues. The results showed that expression of p65 in liver was higher than that in other tissues (P 0.05). After 8-weeks feeding, the hepatic expression level of p65 and IL1β in fish fed diet with high fat showed significant difference with that in control diet (P

Two trials were conducted to determine the effects of dietary macroalgae Porphyra haitanensis on growth, immunity and intestinal microbiota of Litopenaeus vannamei. In trial 1, shrimp (mean initial wet weight about 0.64 g) were fed with seven diets (P0, P1, P2, P3, P4, P5 and P6) containing 0% (basal diet), 1%, 2%, 3%, 4%, 5% and 6% P. haitanensis in triplicate for 60 days. Growth performance (weight gain, WG; specific growth rate, SGR) of shrimp fed the P4 diet were significantly higher than that of shrimp fed P0, P5 and P6 diets (P 0.05) but without significant differences with shrimp fed P1-P3 diets (P 0.05). Hepatopancreas phenoloxidase (PO) activity of shrimp fed the P. haitanensis containing diets was significantly higher than that of shrimp fed the basal diet (P0) (P 0.05). Total haemocyte count (THC) of shrimp fed basal diet (P0) was significantly lower than that of shrimp fed diets containing P. haitanensis. Our results declared that dietary P. haitanensis supplementation increases the abundance of beneficial bacterials such as Nitrosopumilus, Marinobacter or Bifidobacterium and reduces the abundance of harmful bacterias such as Vibrio, and especially pronounced in P4 diet treatment. In trial 2, a WSSV injection challenge test was conducted for 7-day after the rearing trial and shrimp survival was also compared among treatments. A sudden shrimp death was found from the 4th day, and values of survival of shrimp fed the P3-P4 diets were higher than that of shrimp fed other diets during 4-7 days challenge test. The immune response in trial 2 were characterized by higher superoxide dismutase activity (SOD) and PO activities, lower THC and higher HCT compared to levels found in trial 1. In conclusion, suitable dietary P. haitanensis could enhance the growth performance, antioxidant capacity and alter total bacterial numbers or microbial diversity of L. vannamei and furthermore reduce oxidative stress and immune depression challenged by WSSV injection stress, and the level of P. haitanensis supplemented in the diet should be between 2.51% and 3.14%.

Two trials were conducted to determine the effects of dietary Forsythia suspensa extract (FSE) on shrimp, Penaeus monodon, first on growth performance, second on the immune response and immune related gene expression of shrimp. In trial 1, shrimp (mean initial wet weight about 3.02 g) were fed with five diets containing 0% (basal diet), 0.01%, 0.02%, 0.04% and 0.06% FSE in triplicate for 60 days. Growth performance (final body wet weight, FBW; weight gain, WG; biomass gain, BG) of shrimp fed FSE diets were higher (P 0.05) than that of shrimp fed the basal diet. The survival among all the diets treatments were above 90% and no significant difference was revealed among them (P 0.05). The antioxidant capacity (total antioxidant status, TAS; glutathione peroxidase, GSH-Px) appears in the trend of firstly increasing then decreasing with the increasing of dietary FSE levels. The highest value of TAS and GSH-Px were found in shrimp fed 0.02% FSE diet and were significantly higher than that of shrimp fed the basal and 0.06% FSE diets (P 0.05). Hepatopancreas malondialdehyde (MDA) of shrimp fed FSE diets were lower (P 0.05) than that of shrimp fed the basal diet. Total haemocyte count of shrimp fed the basal diet was lower (P 0.05) than that of shrimp fed FSE diets. Haemolymph clotting time of shrimp had the opposite trend with the total haemocyte count of shrimp. No significant differences were found in haemolymph biomarkers of intestinal permeability (endotoxin and diamine oxidase) and in molecular gene expression profiles of heat shock protein 70 (Hsp 70) mRNA and hypoxia inducible factor-1α (HIF-1α) mRNA in haemolymph of shrimp among all diet treatments (P 0.05). In trial 2, a pathogenic strain of Vibrio parahaemolyticus 3HP (VP

The objective of this research was to study the effect of astaxanthin (AST) on growth performance and antioxidant capacity in golden pompano (Trachinotus ovatus) both in vivo and in vitro. In the in vivo study, two diets were formulated with or without astaxanthin supplementation (D1 and D2; 0 and 200 mg/kg) to feed fish for 6 weeks. In the in vitro study, cells from hepatopancreas of golden pompano were isolated and four treatments with or without astaxanthin and H2O2 supplementation were applied (control group: without both astaxanthin and H2O2 treated; H2O2 group: just with H2O2 treated; H2O2 + AST group: with both astaxanthin and H2O2 treated; AST group: just with AST treated). Results of the in vivo study showed that weight gain (WG) and special growth rate (SGR) significantly increased with astaxanthin supplemented (P

To evaluate the utility of the whole-lesion histogram apparent diffusion coefficient (ADC) for characterizing the heterogeneity of mucinous breast carcinoma (MBC) and to determine which ADC metrics may help to best differentiate subtypes of MBC.This retrospective study involved 52 MBC patients, including 37 pure MBC (PMBC) and 15 mixed MBC (MMBC). The PMBC patients were subtyped into PMBC-A (20 cases) and PMBC-B (17 cases) groups. All patients underwent preoperative diffusion-weighted imaging (DWI) at 1.5T and the whole-lesion ADC assessments were generated. Histogram-derived ADC parameters were compared between PMBC vs. MMBC and PMBC-A vs. PMBC-B, and receiver operating characteristic (ROC) curve analysis was used to determine optimal histogram parameters for differentiating these groups.The PMBC group exhibited significantly higher ADC values for the mean (P = 0.004), 25Whole-lesion ADC histogram analysis enables comprehensive evaluation of an MBC in its entirety and differentiating subtypes of MBC. Thus, it may be a helpful and supportive tool for conventional MRI.4 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2018;47:391-400.

Tumor local invasion is the first step of metastasis cascade which remains the key obstacle for cancer therapy. Collective cell migration plays a critical role in tumor invading into surrounding tissues. In vitro assays fail to assess collective invasion in a real time manner. Herein we aim to develop a three-dimensional (3D) microfluidic cell invasion model to determine the dynamic process. In this model, collective invasion of breast cancer cells is induced by the concentration gradient of fetal bovine serum. We find that breast cancer cells adopt a collective movement rather than a random manner when the cells invade into extracellular matrix. The leading cells in the collective movement exhibit an increased expression of an Aurora kinase family protein - AURKA compared with the follower cells. Inhibition of AURKA kinase activity by VX680 or AKI603 significantly reduces the phosphorylation of ERK1/2 (Thr202/Tyr204) and collective cohort formation. Together, our study illustrates that AURKA acts as a potential therapeutic target for suppressing the process of tumor collective invasion. The 3D microfluidic cell invasion model is a reliable, measurable and dynamic platform for exploring potential drugs to inhibit tumor collective invasion.

This study is to explore characteristic indexes in evaluation criteria for rat skin anaphylactoid test comparing skin blue spot OD values at the treated position and the control position in the same animal. Common contrast agents, traditional Chinese medicine injections and injections' active pharmaceutical ingredients or excipients in the existing clinical anaphylactoid reaction reports were taken as test drugs in the rat skin anaphylactoid test to define the K value: K2 represents positive anaphylactoid reaction, 1.2 ≤ K ≤ 2 represent doubtable anaphylactoid; K1.2 represents negative anaphylactoid reaction, which were taken as the criteria for evaluating anaphylactoid of tested drugs. The evaluation result and that for classic criteria were compared to study the applicability of K value. According to the comparison, K value, as the evaluation criteria in the rat skin anaphylactoid test, can more truly reflect the actual situation of skin aizen and minimize the error caused by animal individual factors. Compared with positive and negative two-level criteria for blue spot diameter, K value's positive, doubtable and negative three-level criteria are more objective and accurate. Therefore, K value can be used as the evaluation criteria in the rat skin anaphylactoid test.

Nutritional values and apparent digestibility coefficients for dry matter (ADMD), protein (APD), energy (AED) and amino acid (AAAD) of ten feedstuffs were determined for juvenile shrimp, which included Peruvian steam fish meal, Peruvian FAQ fish meal, local fish meal, soybean meal, peanut meal, shrimp head meal, cottonseed meal, rapeseed meal, wheat flour and brewer’s yeast. A reference diet and test diets that consisted of a 70:30 mixture of the reference diet to test feedstuff were used with 0.01% Yttrium oxide as an external marker. The eleven diets were fed to triplicate groups of juvenile Penaeus monodon (mean initial weight 1.13 g ± 0.02 g) for 56 days. After the 56-day trial, the highest and lowest values of weight gain (WG) and specific growth rate (SGR) of shrimp were found in shrimp head meal and cottonseed meal diet treatments, respectively. Feed conversion ratio (FCR) was in the range of 1.25-1.65. Peruvian steam fish meal and shrimp head meal diet treatments had the lowest FCR and cottonseed meal and rapeseed meal diet treatments had the highest FCR. The ADMD of diets is in the range of 63.52% to 80.07%. The values of ADMD in shrimp head meal and wheat flour diets (77.58% and 80.07%) were significantly higher than that in other diets (P

This study aims to evaluate the multidetector computed tomography (CT) imaging features in differentiating exophytic renal angiomyolipoma (AML) from retroperitoneal liposarcoma. We retrospectively enrolled 42 patients with confirmed exophytic renal AML (31 patients) or retroperitoneal liposarcoma (11 patients) during 8 years period to assess: renal parenchymal defect at site of tumor contact, supply from branches of renal artery, tumoral vessel extending through the renal parenchyma, dilated intratumoral vessels, hemorrhage, non-fat-containing intratumoral nodules with postcontrast enhancement, calcification, renal sinus enlargement, anterior displacement of kidneys, and other associated AML. Renal parenchymal defect, renal arterial blood supply, tumoral vessel through the renal parenchyma, dilated intratumoral vessels, intratumoral/perirenal hemorrhage, renal sinus enlargement, and associated AML were seen only or mainly in exophytic renal AML (all P value