A proteomic study on gastric impairment in rats caused by microcystin-LR.

Microcystins (MCs) are the most common cyanobacterial toxins. Epidemiological investigation showed that exposure to MCs can cause gastro-intestinal symptoms, gastroenteritis and gastric cancer. MCs can also accumulate in and cause histopathological damage to stomach. However, the exact mechanisms by which MCs cause gastric injury were unclear. In this study, Wistar rats were administrated 50, 75 or 100 µg microcystin-LR (MC-LR)/kg, body mass (bm) via tail vein, and histopathology, response of anti-oxidant system and the proteome of gastric tissues at 24 h after exposure were studied. Bleeding of fore-stomach and gastric corpus, inflammation and necrosis in gastric corpus and exfoliation of mucosal epithelial cells in gastric antrum were observed following acute MC-LR exposure. Compared with controls, activities of superoxide dismutase (SOD) were significantly greater in gastric tissues of exposed rats, while activities of catalase (CAT) were less in rats administrated 50 µg MC-LR/kg, bm, and concentrations of glutathione (GSH) and malondialdehyde (MDA) were greater in rats administrated 75 or 100 µg MC-LR/kg, bm. These results indicated that MC-LR could disrupt the anti-oxidant system and cause oxidative stress. The proteomic results revealed that MC-LR could affect expressions of proteins related to cytoskeleton, immune system, gastric functions, and some signaling pathways, including platelet activation, complement and coagulation cascades, and ferroptosis. Quantitative real-time PCR (qRT-PCR) analysis showed that transcriptions of genes for ferroptosis and gastric function were altered, which confirmed results of proteomics. Overall, this study illustrated that MC-LR could induce gastric dysfunction, and ferroptosis might be involved in MC-LR-induced gastric injury. This study provided novel insights into mechanisms of digestive diseases induced by MCs.

Development a novel nomogram model for predicting significant hepatic histological changes in chronic hepatitis B virus carriers.

A proportion of chronic hepatitis B virus (HBV) carriers with normal alanine transaminase (ALT) present with significant liver histological changes (SLHC). To construct a noninvasive nomogram model to identify SLHC in chronic HBV carriers with different upper limits of normal (ULNs) for ALT. The training cohort consisted of 732 chronic HBV carriers who were stratified into four sets according to different ULNs for ALT: chronic HBV carriers I, II, III, and IV. The external validation cohort comprised 277 chronic HBV carriers. Logistic regression and least absolute shrinkage and selection operator analyses were applied to develop a nomogram model to predict SLHC. A nomogram model-HBGP (based on hepatitis B surface antigen, gamma-glutamyl transpeptidase, and platelet count) demonstrated good performance in diagnosing SLHC with area under the curve (AUCs) of 0.866 (95% confidence interval [CI]: 0.839-0.892) and 0.885 (95% CI: 0.845-0.925) in the training and validation cohorts, respectively. Furthermore, HBGP displayed high diagnostic values for SLHC with AUCs of 0.866 (95% CI: 0.839-0.892), 0.868 (95% CI: 0.838-0.898), 0.865 (95% CI: 0.828-0.901), and 0.853 (95% CI: 0.798-0.908) in chronic HBV carriers I, II, III, and IV, respectively. Additionally, HBGP showed greater ability in predicting SLHC compared with the existing predictors. HBGP has shown high predictive performance for SLHC, and thus may lead to an informed decision on the initiation of antiviral treatment.

In Situ-Formed Fibrin Hydrogel Scaffold Loaded With Human Umbilical Cord Mesenchymal Stem Cells Promotes Skin Wound Healing.

Healing of full-thickness skin wounds remains a major challenge. Recently, human umbilical cord mesenchymal stem cells (hUC-MSCs) were shown to possess an extraordinary potential to promote skin repair in clinical settings. However, their low survival rate after transplantation limits their therapeutic efficiency in treating full-thickness skin wounds. Hydrogels are considered an ideal cell transplantation vector owing to their three-dimensional mesh structure, good biosafety, and biodegradation. The objective of this study was to investigate the skin wound healing effect of a fibrin hydrogel scaffold loaded with hUC-MSCs. We found that the fibrin hydrogel had a three-dimensional mesh structure and low cytotoxicity and could prolong the time of cell survival in the peri-wound area. The number of green fluorescent protein (GFP)-labeled hUC-MSCs was higher in the full-thickness skin wound of mice treated with hydrogel-hUC-MSCs than those of mice treated with cell monotherapy. In addition, the combination therapy between the hydrogel and hUC-MSCs speed up wound closure, its wound healing rate was significantly higher than those of phosphate-buffered saline (PBS) therapy, hydrogel monotherapy, and hUC-MSCs monotherapy. Furthermore, the results showed that the combination therapy between hydrogel and hUC-MSCs increased keratin 10 and keratin 14 immunofluorescence staining, and upregulated the relative gene expressions of epidermal growth factor (EGF), transforming growth factor-ß1 (TGF-ß1), and vascular endothelial growth factor A (VEGFA), promoting epithelial regeneration and angiogenesis. In conclusion, the fibrin hydrogel scaffold provides a relatively stable sterile environment for cell adhesion, proliferation, and migration, and prolongs cell survival at the wound site. The hydrogel-hUC-MSCs combination therapy promotes wound closure, re-epithelialization, and neovascularization. It exhibits a remarkable therapeutic effect, being more effective than the monotherapy with hUC-MSCs or hydrogel.

Characterization of complete mitochondrial genome of the common tiger, Danaus genutia (Lepidoptera: Nymphalidae: Danainae) and phylogenetic implications within the subfamily Danainae.

Nymphalidae is the most diverse butterfly family worldwide, with more than 6000 species, whereas the mitogenomic data of nymphalid species, especially the subfamily Danainae, is still lacking for more comprehensive systematic studies. To this contribution, the complete mitogenome of Danaus genutia (Lepidoptera: Nymphalidae: Danainae) was determined via sequencing and annotating. The mitogenome in total consists of 15,255 base pairs (bp), containing 13 protein-coding genes, 22 transfer RNAs, 2 ribosomal RNAs, and a 440-bp noncoding A+T-rich region. Furthermore, phylogeny of the subfamily Danainae was reconstructed applying maximum likelihood and Bayesian inference on the basis of the mitogenomic data sets. Combined with our analysis and previous studies, the genus-level phylogenetic relationships within the subfamily Danainae are ((Tirumala + Danaus) + ((Idea + Euploea) + (Ideopsis + Parantica))). This study offers molecular information and provides a new perspective for phylogenetic research within the subfamily Danainae.

Dual-Function Analysis of Astaxanthin on Golden Pompano (Trachinotus ovatus) and Its Role in the Regulation of Gastrointestinal Immunity and Retinal Mitochondrial Dysfunction Under Hypoxia Conditions.

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-3ß (pGSK-3ß), 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 rhodamine-based dual chemosensor for the naked-eye detection of Hg2+ and enhancement of the fluorescence emission for Fe3.

A novel fluorescent chemosensor based on trimesoyl chloride-rhodamine (TR) was successfully synthesized. Rising chromogenic and fluorogenic spectral enhancements could be observed in trimesoyl chloride-rhodamine (TR) probes when Hg2+ and Fe3+ were added, respectively. TR has shown selectivity for Hg2+ and Fe3+ with high sensitivity due to metal ion complexation induced photophysical "turn-on" signaling responses. The detection limit towards Hg2+ was 2.46 × 10-8 M as determined by the 3σ method. At the same time, fluorogenic spectral enhancements were observed in TR, which exhibits a superior sensitive and selective recognition towards Fe3+ with 4.11 × 10-8 M of the detection limit. The test strips were used for colorimetric and simple detection towards Hg2+, which might finally enable the advancement of the Hg2+ sensor in the field of on-site detection.

A fluorescent supramolecular gel and its application in the ultrasensitive detection of CN- by anion-π interactions.

Supramolecular gels have been widely reported on account of their unique superiority and application prospects. In this work, we constructed a novel supramolecular gel (HD-G) by using hydroxy-naphthaldehyde decorated with naphthalimide in DMSO solution, which exhibited excellent selectivity and ultrasensitive sensing properties toward CN- (the lowest detection limit is 1.82 × 10-10 M). The sensing mechanism of this supramolecular gel takes advantage of π-π stacking interactions and anion-π interactions, which is different from the other familiar methods.

The sex- and age-specific association between resting heart rate and hypertension in rural adult residents of Henan province / 中华心血管病杂志

Objective: To investigate the sex- and age-specific association between resting heart rate and hypertension in rural adult residents of Henan province. Methods: At baseline, a total of 20 194 participants were randomly selected from Xin'an County of Henan province between July 2007 and August 2008. After excluding participants with hypertension or without resting heart rate data at baseline, and participants died or without hypertension outcome or diagnosed as gestational hypertension during follow-up between July 2013 and October 2014, 10 212 participants were finally included in this study. Multiple linear regression model was used to examine the association between resting heart rate and change of blood pressure. Logistic regression model was used to estimate the association between resting heart rate and risk of hypertension. Results: There were 2 059 new hypertensive cases (839 male) during the 6 years follow-up. After controlling for potential confounders, per 5 beats/minutes increases in resting heart rate was associated with 0.18 mmHg (1 mmHg=0.133 kPa) (95%CI 0.01-0.36 mmHg, P=0.046) absolute increase in systolic blood pressure and 7% higher risk of developing hypertension in women (95%CI 1.03-1.11, P<0.05). Compared with resting heart rate<70 beats/minutes, the adjusted RRs for 76-82 and>82 beats/minutes groups were 1.39 (95%CI 1.18-1.63, P<0.05) and 1.22 (95%CI 1.02-1.45, P<0.05), respectively. For both age groups, increased resting heart rate was positively associated with risk of hypertension in women(RR=1.05(95% CI 1.01-1.10), P<0.05 (the women those <60 years); RR=1.14(95% CI 1.04-1.25), P<0.05 (the women those≥60 years). However, no significant association was found between resting heart rate and hypertension in male residents. Conclusions: Increased resting heart rate is associated with high risk of hypertension in women who live in rural area, especially in elder women of this cohort.

The correlation between the methylation of PTEN gene and the apoptosis of osteosarcoma cells mediated by SeHA nanoparticles.

The invasive spreading of residual osteosarcoma cells becomes a serious threat to human health, urgently needing new bone regenerative biomaterials for orthopedic therapy. Thus, in this work, selenite-substituted hydroxyapatite (SeHA) nanoparticles were prepared for both inhibiting the recurrence of the tumor and accelerating the regenerative repair of bone defect. Physicochemical characterization showed these synthetic nanoparticles were spherical poly-crystals with the shape of snowflakes. Such structure benefited them to inhibit the cellular viability of osteosarcoma cells by about (58.90 ± 14.37)% during 24 h co-culturing. The expression level of cell growth-related genes such as PTEN, MMP-9, Cyclin D1, Cyclin A2, Annexin A2 and CDC2 decreased. Bisulfite Sequence PCR of PTEN gene exhibited about (22.40 ± 5.39)%, (45.91 ± 6.36)% and (25.90 ± 5.36)% promoter methylation in control, HA and SeHA group. Animal experiment also proved the similar effects. Almost no recurrence were observed in SeHA group. Oppositely, the slowly recurrent growth of the remnant tumor appeared in purely surgical group. The overall survival and toxicity analysis showed that, in the usage dose of 0-0.1 g, the SeHA-0.01 exhibited higher inhibitory recurrence and metastasis potentials, lower renal toxicity and better anti-inflammation function. Immunohistochemistry stain showed the reduced expression of PTEN, MMP-9, Ki-67 and Annexin A2, but slightly increased expression of DNMT1 and BMP-2. Compared the methylation status of PTEN gene in each group, it was confirming that SeHA nanoparticles hardly possessed the de-methylation effect, but the pure HA strikingly increased the methylation level of such gene. It seemed the dopant selenite ions possessed de-methylation effect onto PTEN gene. Therefore, from the viewpoint of inhibiting metastatic potentials, the SeHA-0.01 might be a feasible biomaterial to inhibit the relapse of the tumor post-surgery.

The reduction of XIAP is associated with inflammasome activation in RPE: implications for AMD pathogenesis.

BACKGROUND: Age-related macular degeneration (AMD) is a multifactorial chronic disease of the eye. Several candidate pathways have been hypothesized to play a role in AMD pathogenesis. Our work and those of others suggests inflammasome activity as a mechanism associated with retinal pigment epithelial (RPE) cell demise. X-linked inhibitor of apoptosis protein (XIAP), an anti-apoptosis factor, has recently been shown to regulate inflammasome activity in non-ocular cells. The purpose of this study is to characterize XIAP's regulatory role in RPE. METHODS: Protein lysates of eye tissues from rats (vinpocetine- or aurin tricarboxylic acid complex-treated, ATAC, vs naïve) and mice (wild type vs Caspase-4-/-) were utilized to analyze XIAP protein levels. Immunohistochemistry was used to detect NLRP3 levels in the RPE layer. In vitro inflammasome activation on RPE cells was achieved with L-leucyl-L-leucine methyl ester (Leu-Leu-OMe) stimulation. Levels of XIAP mRNA and 18S RNA were quantified by RT-PCR. Cell culture supernatants were tested directly for secreted IL-1ß by ELISA or concentrated for the detection of secreted IL-18 by western blot. Protein lysates from RPE in cell culture were collected for the measurement of cleaved caspase-1 p20, XIAP, and GAPDH. Data are presented as Mean ± SD. p < 0.05 is considered statistically significant. RESULTS: The XIAP protein level was significantly increased when the inflammasome was inhibited at the "activation" step by ATAC, but not the "priming" step, in vivo. Concomitantly, NLRP3 immunoreactivity was lower in the RPE layer of animals fed with ATAC. In mice where caspase-1 cleavage was impaired by the genetic deficiency in caspase-4, the XIAP protein level increased in eye tissues. In RPE cell culture, Leu-Leu-OMe stimulation led to caspase-1 cleavage, cytokine secretion, and XIAP reduction, which can be abolished by Z-YVAD-FMK. When XIAP siRNA was given as a pre-treatment to RPE in vitro, Leu-Leu-OMe induced IL-1ß/IL-18 secretion was enhanced, whereas overexpressing XIAP reduced IL-1ß secretion under inflammasome activation, both compared to controls cells. CONCLUSIONS: Together, these data suggest XIAP-mediated inhibition of inflammasome activity in RPE may provide insights into the biological consequences of inflammasome activation in RPE and reveals the caspase-1/XIAP/IL-1ß/IL-18 axis as a target for broader applications in AMD biology and treatment design.

Effects of dietary mixed probiotics on growth, non-specific immunity, intestinal morphology and microbiota of juvenile pacific white shrimp, Litopenaeus vannamei.

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 ±â€¯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.

Chocolate consumption and risk of cardiovascular diseases: a meta-analysis of prospective studies.

OBJECTIVE: Studies investigating the impact of chocolate consumption on cardiovascular disease (CVD) have reached inconsistent conclusions. As such, a quantitative assessment of the dose-response association between chocolate consumption and incident CVD has not been reported. We performed a systematic review and meta-analysis of studies assessing the risk of CVD with chocolate consumption. METHODS: PubMed and EMBASE databases were searched for articles published up to 6 June 2018. Restricted cubic splines were used to model the dose-response association. RESULTS: Fourteen publications (23 studies including 405 304 participants and 35 093 cases of CVD) were included in the meta-analysis. The summary of relative risk (RR) per 20 g/week increase in chocolate consumption was 0.982 (95% CI 0.972 to 0.992, I2=50.4%, n=18) for CVD (heart failure: 0.995 (0.981 to 1.010, I2=36.3%, n=5); total stroke: 0.956 (0.932 to 0.980, I2=25.5%, n=7); cerebral infarction: 0.952 (0.917 to 0.988, I2=0.0%, n=4); haemorrhagic stroke: 0.931 (0.871 to 0.994, I2=0.0%, n=4); myocardial infarction: 0.981 (0.964 to 0.997, I2=0.0%, n=3); coronary heart disease: 0.986 (0.973 to 0.999, n=1)). A non-linear dose-response (pnon-linearity=0.001) indicated that the most appropriate dose of chocolate consumption for reducing risk of CVD was 45 g/week (RR 0.890;95%CI 0.849 to 0.932). CONCLUSIONS: Chocolate consumption may be associated with reduced risk of CVD at <100 g/week consumption. Higher levels may negate the health benefits and induce adverse effects associated with high sugar consumption.

Assembly Mechanism of Highly Crystalline Selenium-Doped Hydroxyapatite Nanorods via Particle Attachment and Their Effect on the Fate of Stem Cells.

Hydroxyapatite is the main inorganic component of human bone. Synthetic hydroxyapatite and its different modified forms, which have been shown to be biocompatible and osteoconductive, have been widely used in bone tissue engineering. It is still challenging to controllably synthesize hydroxyapatite with a targeted morphology. In this work, we synthesized highly crystalline selenium-doped hydroxyapatite nanorods (SeHAN) via a two-step alcohol thermal method and provided a complete explanation of the synthesis mechanism. Tracing the crystals obtained from the solvated phase to the crystal phase with high-resolution microscopy, the nanorod formation route can be briefly described as follows: as a basic unit, ∼30 nm amorphous apatite initially formed in the first step and partly crystallized in early part of the second step; after a period of alcohol thermal reaction, immature nanorods appeared, which were composed of nanocrystals; finally, immature nanorods transformed into single-crystal nanorods through crystallization by particle attachment. Since few works have focused on the osteogenesis ability of SeHAN, whose antitumor effect has been widely studied, we investigated the influences of SeHAN on rat-bone-marrow-resident mesenchymal stem cells (MSCs). Surprisingly, SeHAN exhibited excellent biocompatibility for MSCs, enhanced their osteogenic differentiation, and inhibited their adipogenic differentiation. This work provides not only a general method to controllably synthesize hydroxyapatite nanorods/SeHAN but also an insight into understanding the hydroxyapatite formation mechanism. The current study also highlights the effects of SeHAN on MSCs that could furnish a significative strategy for manufacturing functional biomaterials for biomedical applications and tissue engineering by enhanced ossification and reduced marrow adiposity.

Dietary values of macroalgae Porphyra haitanensis in Litopenaeus vannamei under normal rearing and WSSV challenge conditions: Effect on growth, immune response and intestinal microbiota.

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%.

[Mechanism of Action of Activated Sludge Properties in Nitrogen Removal by Endogenous Denitrification Through an Intelligent Aeration-controlled A/O Process].

In order to evaluate the mechanism of action of activated sludge properties in nitrogen removal by endogenous denitrification (henceforth EDNR), a new kind of automatic oxygen supply device(AOSD), was applied to the A/O process. The domestication effect of the aeration mode on the activated sludge properties and microbial communities was investigated under the intelligent aeration-controlled A/O process (I-A/O)and the continuous aeration A/O process (C-A/O). The results demonstrated that the effluent NH4+-N and NO2--N components showed obvious accumulation efficiencies and activated sludge generated conspicuous limited bulking in the I-A/O process. Domesticated sludge in the I-A/O process was able to enrich more SCOD to transfer into the polymeric substances as Gly, under a rich exogenous carbon supply state, and stimulated nitrogen removal by endogenous denitrifying under a scarce exogenous carbon supply state. The EDNR rate went up to 0.83 mg·(L·h)-1 in the I-A/O process, which was more than that achieved by the C-A/O process. The microbial communities in the two processes were evaluated by the Illumina HiSeq high-throughput sequencing technology. The results showed that there was no obvious difference in the sludge microbial community diversity between the two processes, but the Candidate division TM7 proliferated in the I-A/O process, and become the abundant taxa to prompt limited filamentous sludge bulking and Gly storage capability enhancement. The oxygen supply mode of AOSD made the activated sludge properties and microbial communities to be screened selectively in the new environment, aerobic heterotrophic bacterial activity to decline, and endogenous denitrifying action to strengthen, which made the I-A/O process implement a kind of dynamic balanced state that limited the DO demand.

Dietary values of Forsythia suspensa extract in Penaeus monodon under normal rearing and Vibrio parahaemolyticus 3HP (VP3HP) challenge conditions: Effect on growth, intestinal barrier function, immune response and immune related gene expression.

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 (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). Dietary FSE supplementation produced stronger hepatopancreas antioxidant capacity (TAS, GSH-Px) (P < 0.05) and higher glutathione (GSH) level (P < 0.05), lower superoxide dismutase activity (SOD) (P < 0.05), higher total haemocyte count (P < 0.05), lower haemolymph clotting time (P < 0.05), lower MDA and carbonyl protein concentration (P < 0.05), lower haemolymph biomarkers of intestinal permeability (endotoxin and diamine oxidase) (P < 0.05), generated lower molecular gene expression profiles of HSP 70 mRNA and higher HIF-1α mRNA (P < 0.05) than the basal diet. The immune response were characterized by lower TAS and higher antioxidant enzyme activities (SOD, GSH-Px) and higher oxidative stress level (MDA and carbonyl protein) and higher haemolymph biomarkers of intestinal permeability (endotoxin and diamine oxidase) compared to levels found in trail 1. However, the total haemocyte counts and haemolymph clotting times were not changed in 0.01%-0.02% FSE diets treatments between trial 1 and trial 2 (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%.

Comparison and Evaluation of Four Species of Macro-Algaes as Dietary Ingredients in Litopenaeus vannamei Under Normal Rearing and WSSV Challenge Conditions: Effect on Growth, Immune Response, and Intestinal Microbiota.

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 ß. 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.

Biomimetic Coprecipitation of Silk Fibrin and Calcium Phosphate: Influence of Selenite Ions.

Large bone defect creates an urgent need for osteogenic biomaterials. However, bone nonunion and infection are choke points in the therapy of this disease. How to recruit the mesenchymal stem cells to defect sites and increase the cell viability are the critical processes. One effective method was the fabrication of biomimetic silk fibrin/selenium-doped hydroxyapatite (SF/HASe) material, which could create a niche for cell proliferation. So, the aim of the present study was to seek a facile route to prepare this biocomposites and investigate the osteogenic capability. Results showed that the biomimetic coprecipitation was a successful route to prepare SF/HASe biocomposites, which presented higher cell proliferation activity and better modulation of the selenite release during incubation in biological medium. Besides, the biocomposites exhibited weird and porous pot morphology. Such features could provide large surface area for the cells and proteins to attach. Silk fibrin, adhered onto the surface of hydroxyapatite (HA) crystals, plays a crucial impact on the release profile of selenite ions. The release behavior of the selenite ions exhibited stably slow release fashion. Therefore, it is feasible to employ SF/HASe biocomposites to repair bone defects and apply into the therapy of osteosarcoma postoperatively.

In Vitro and in Vivo Mechanism of Bone Tumor Inhibition by Selenium-Doped Bone Mineral Nanoparticles.

Biocompatible tissue-borne crystalline nanoparticles releasing anticancer therapeutic inorganic elements are intriguing therapeutics holding the promise for both tissue repair and cancer therapy. However, how the therapeutic inorganic elements released from the lattice of such nanoparticles induce tumor inhibition remains unclear. Here we use selenium-doped hydroxyapatite nanoparticles (Se-HANs), which could potentially fill the bone defect generated from bone tumor removal while killing residual tumor cells, as an example to study the mechanism by which selenium released from the lattice of Se-HANs induces apoptosis of bone cancer cells in vitro and inhibits the growth of bone tumors in vivo. We found that Se-HANs induced apoptosis of tumor cells by an inherent caspase-dependent apoptosis pathway synergistically orchestrated with the generation of reactive oxygen species. Such mechanism was further validated by in vivo animal evaluation in which Se-HANs tremendously induced tumor apoptosis to inhibit tumor growth while reducing systemic toxicity. Our work proposes a feasible paradigm toward the design of tissue-repairing inorganic nanoparticles that bear therapeutic ions in the lattice and can release them in vivo for inhibiting tumor formation.

Changes in serum adipocyte fatty acid-binding protein in women with gestational diabetes mellitus and normal pregnant women during mid- and late pregnancy.

AIMS/INTRODUCTION: To observe the longitudinal changes in serum adipocyte fatty acid-binding protein (AFABP), carbohydrate, and lipid metabolism parameters in women with and without gestational diabetes mellitus (GDM) during mid- and late pregnancy periods, as well as to identify whether there is any association between AFABP and development of GDM. MATERIALS AND METHODS: A total of 40 GDM and 240 normal glucose tolerance participants were enrolled at 24-28 weeks and completed the study. The clinical features, serum AFABP, other adipocytokines (leptin, adiponectin, retinol-binding protein 4), homeostasis model assessment of insulin resistance, and lipid profiles were measured in the second and third trimesters of pregnancy. RESULTS: Compared with the normal glucose tolerance group, the GDM group showed greater levels of AFABP, leptin and retinol-binding protein 4; and a decreased level of adiponectin (P < 0.05 or P < 0.01) during mid- and late pregnancy periods. Prepregnancy body mass index was the independent factor impacting serum AFABP levels in the second (ß = 0.567, P = 0.004) and third trimesters (ß = 0.619, P = 0.001). Furthermore, GDM was independently associated with AFABP concentrations in multiple regression analysis in the second and third trimester (all P < 0.01). Serum AFABP, leptin and retinol-binding protein 4 are risk factors for GDM; adiponectin is a protective factor for GDM (P < 0.05 or P < 0.01). CONCLUSIONS: The GDM group had a higher level of AFABP during mid- and late stages of pregnancy; prepregnancy body mass index and GDM were the independent factors with respect to serum AFABP. AFABP might be closely related to obesity, insulin resistance and leptin resistance in pregnancy, and is a major risk factor for GDM.