Effects of Continuous Quality Improvement on Complication Rate and Treatment Outcome of Extremely Premature Infants.

Background: Extremely premature infants (EPIs) are those less than 32 weeks of gestational age. Preterm birth is the leading cause of neonatal death and poor prognosis, accounting for 25% of neonatal deaths, with extremely premature births accounting for 50% of all premature deaths. Continuous quality improvement (CQI) improves patient outcomes by changing and optimizing clinical practice including increasing participation of neonatologists in prenatal consultation, maintenance of normal body temperature in preterm infants, early use of pulmonary surfactant, reduction of mechanical ventilation time and intensive breastfeeding to reduce clinically avoidable adverse events. Objective: The risk of death and disability is high for very preterm infants, with a mortality rate of 30-50% and a risk of at least 20-50% for survivors. This study aimed to investigate the effect of CQI on the incidence of complications and treatment outcomes in very preterm infants. Design: This was a retrospective study. Setting: This study was conducted in the Maternal and Child Health Hospital of Hubei Province. Participants: A total of 140 EPIs born in our hospital and transferred to the neonatal intensive care unit between August 1, 2020, and July 31, 2022, were enrolled. The EPIs were divided into two groups: before improvement (n=79, 56.4%) and after improvement (n=61, 43.6%) according to the week of birth, and the gestational age ranged from 26 weeks to 26 weeks 6 days into the 26 weeks group. Interventions: From August 2021, the hospital implemented the CQI method, which included neonatologists' participation in consultations before birth, the care of a professionally trained resuscitation team after birth, and the introduction of transport heating tanks and ventilators during transport. Primary Outcome Measures: (1) Apgar score (2) body weight (3) duration of invasive ventilation (4) length of stay (5) treatment expense (6) incidence of complications and (7) survival rate of EPIs. Results: The application of CQI methods resulted in significant improvements in body weight (1305 g vs 1404 g) and duration of invasive ventilation (4.64 d vs 7.40 d) in EPIs (P = .036 and P = .040), reduced the time of invasive mechanical ventilation decreased significantly, from 7.4 days to 4.64 days (P < .01), increased the median temperature of newborn infants (36.2°C vs 35.7°C) (P = 0), increased the proportion of newborn infants with a temperature greater than 36°C (67.2% vs 35.4%) (P < .001), reduced the incidence of complications in EPIs (32.79% vs 45.57%) (P < .05). Conclusion: The application of the CQI approach significantly increases the body temperature, improves the incidence of complications of EPIs, and is conducive to the survival of EPIs. Our study may provide a clinical reference for management of EPIs.

Metabolism response mechanism in the gill of Oreochromis mossambicus under salinity, alkalinity and saline-alkalinity stresses.

Saline-alkalinity is one of the important ecological parameter that has an impact function on the physiological metabolism, osmoregulation, survival, growth, development and distribution of teleost fish. Oreochromis mossambicus, a species of euryhaline that can withstand a wide variety of salinities, may be used as a research model animal in environmental studies. In order to detect the metabolism responses and mechanisms of different osmotic stresses tolerance in the gills of O. mossambicus, in present study, the metabolic responses of O. mossambicus subjected to salinity (25 g/L, S_S), alkalinity (4 g/L, A_S) and saline-alkalinity stress (salinity: 25 g/L, alkalinity: 4 g/L; SA_S) with the control environment (freshwater, C_S) were investigated by LC-MS/MS-based metabolomics. The metabolism results indicated that numerous metabolites were identified between the stress groups and the control group. In addition, under three osmotic stresses, the amino acid and carbohydrate metabolism, levels of amino acids, osmolytes and energy substances, such as L-lysine, arachidonic acid, docosahexaenoic acids, creatine and taurine, were significantly affected and changed in the metabolism of the gills of O. mossambicus. The metabolism data indicated that signal transduction and regulation pathways, including FoxO signaling pathway, mTOR signaling pathway and prolactin signaling pathway, were enriched in the gill during adaptation to high salinity, alkalinity and saline-alkalinity stress. The results of this study provide more comprehensive and reliable data for the osmotic pressure regulation mechanism and biological response of euryhaline teleost, and provide reliable scientific basis for the breeding and research of high salinity tolerance population, and further promote the development and utilization of saline-alkalinity water resources.

Effect of lipase incorporation on gelling properties of catfish (Clarias lazera) surimi and its mechanism.

BACKGROUND: Recently, fatty fish have been utilized as a potential approach for the fabrication of surimi products, with the yield of fatty fish surimi being > 10 000 tons in 2019. However, the gelling properties of catfish surimi can be influenced by intermuscular lipid. Lipase could effectively enhance the gel quality of catfish surimi gels, although the chemical forces involved in gel formation and alteration in lipid and protein oxidation status are not well understood. The present study investigated the gelation-enhancing effects of lipase on catfish surimi based on changes in chemical oxidation interactions. RESULTS: The addition of 7.5 g kg-1 lipase significantly increased the hydrophobic interactions and disulfide bond contents, both of which facilitated gel formation, in surimi gels. The 2-thiobarbituric acid reactive substance and carbonyl concentrations demonstrated that lipase promoted lipid and protein oxidations. Furthermore, an appropriate dose of malondialdehyde accelerated protein oxidation, thereby resulting in the covalent cross-linking of proteins. Consequently, the gel strength increased from 55.72 to 127.71 g × cm with lipase contents of up to 7.5 g kg-1 , and strong chemical cross-linking and a compact network were observed via sodium dodecyl sulfate polyacrylamide gel electrophoresis and scanning electron microscopy. However, excessive oxidation led to the degeneration of the gel matrix. A schematic mechanism, mainly based on the chemical changes, is proposed. CONCLUSION: The present study revealed the gelation mechanism of catfish surimi gels with lipase, and suggested that lipase treatments may be an effective approach for improving the textural properties of fatty fish surimi gels. © 2021 Society of Chemical Industry.

Transcriptomic response to three osmotic stresses in gills of hybrid tilapia (Oreochromis mossambicus female × O. urolepis hornorum male).

BACKGROUND: Osmotic stress is a widespread phenomenon in aquatic animal. The ability to cope with salinity stress and alkaline stress is quite important for the survival of aquatic species under natural conditions. Tilapia is an important commercial euryhaline fish species. What's more tilapia is a good experimental material for osmotic stress regulation research, but the molecular regulation mechanism underlying different osmotic pressure of tilapia is still unexplored. RESULTS: To elucidate the osmoregulation strategy behind its hyper salinity, alkalinity and salinity-alkalinity stress of tilapia, the transcriptomes of gills in hybrid tilapia (Oreochromis mossambicus ♀ × O. urolepis hornorum ♂) under salinity stress (S: 25‰), alkalinity stress(A: 4‰) and salinity-alkalinity stress (SA: S: 15‰, A: 4‰) were sequenced using deep-sequencing platform Illumina/HiSeq-2000 and differential expression genes (DEGs) were identified. A total of 1958, 1472 and 1315 upregulated and 1824, 1940 and 1735 downregulated genes (P-value < 0.05) were identified in the salt stress, alkali stress and saline-alkali stress groups, respectively, compared with those in the control group. Furthermore, Kyoto Encyclopedia of Genes and Genomes pathway analyses were conducted in the significant different expression genes. In all significant DEGs, some of the typical genes involved in osmoregulation, including carbonic anhydrase (CA), calcium/calmodulin-dependent protein kinase (CaM kinase) II (CAMK2), aquaporin-1(AQP1), sodium bicarbonate cotransporter (SLC4A4/NBC1), chloride channel 2(CLCN2), sodium/potassium/chloride transporter (SLC12A2 / NKCC1) and other osmoregulation genes were also identified. RNA-seq results were validated with quantitative real-time PCR (qPCR), the 17 random selected genes showed a consistent direction in both RNA-Seq and qPCR analysis, demonstrated that the results of RNA-seq were reliable. CONCLUSIONS: The present results would be helpful to elucidate the osmoregulation mechanism of aquatic animals adapting to saline-alkali challenge. This study provides a global overview of gene expression patterns and pathways that related to osmoregulation in hybrid tilapia, and could contribute to a better understanding of the molecular regulation mechanism in different osmotic stresses.

Selection of reference genes for quantitative real-time PCR normalisation in largemouth bass Micropterus salmoides fed on alternative diets.

The partial cDNA sequences of eight reference genes (actb, tuba1, gapdh58, gapdh59, eef1a1, RNA 18 s, pabpc1, ube2I) were cloned from largemouth bass Micropterus salmoides. The expression levels of these eight genes were compared in the various tissues (eye, spleen, kidney, gill, muscle, brain, liver, heart, gut and gonad) of M. salmoides fed on forage fish. The results showed that the candidate genes exhibited tissue-specific expression to various degrees and the stability ranking order was eef1a1 > tuba1 > RNA 18 s > pabpc1 > ube2I > actb > gapdh58 > gapdh59 among tissue types. Four candidate genes eef1a1, tuba1, RNA 18 s and actb were used to analyse the stability in liver tissues of largemouth bass between the forage-fish group and the formulated-feed group. The candidate genes also showed some changes in expression levels in the livers, while eef1a1 and tuba1 had the most stable expression in livers of fish fed on alternative diets within 10 candidates. So eef1a1 and tuba1 were recommended as optimal reference gene in quantitative real-time PCR analysis to normalise the expression levels of target genes in tissues and lives of the M. salmoides fed on alternative diets. In livers, the expression levels of gck normalised by eef1a1 and tuba1 showed the significant up-regulation in formulated feed group (P < 0.05) than those in forage-fish group. While sex difference has no significant effects on the expression levels of gck in both groups.

Effects of dietary Lactobacillus rhamnosus JCM1136 and Lactococcus lactis subsp. lactis JCM5805 on the growth, intestinal microbiota, morphology, immune response and disease resistance of juvenile Nile tilapia, Oreochromis niloticus.

The present study aimed to evaluate the individual and combined effects of Lactobacillus rhamnosus (LR) JCM1136 and Lactococcus lactis subsp. lactis (LL) JCM5805 on the growth, intestinal microbiota, intestinal morphology, immune response and disease resistance of juvenile Nile tilapia (Oreochromis niloticus). A total of 720 apparently healthy juvenile Nile tilapia (0.20 ± 0.05 g) were randomly divided into four equal groups. Fish were fed with a basal diet (CK) supplemented with JCM1136 (LR), JCM5805 (LL), and JCM1136 + JCM5805 (LR+LL) at 1 × 108 CFU/g basal diet for 6 weeks, followed by a basal diet for 1 week. After 6 weeks of feeding, the LL treatment significantly increased the growth and feed utilization of Nile tilapia when compared with the CK. Light microscopy and transmission electron microscopy images of the midgut revealed that probiotic supplementation significantly increased gut microvilli length and microvilli density compared to CK. The transcript levels of several key immune-related genes in the mid-intestine and liver of fish were analyzed by means of quantitative polymerase chain reaction (qPCR) at the end of the sixth week. The results showed the following: when compared to CK group, fish in LR had significantly increased transcript levels of IFN-γ, lyzc, hsp70 and IL-1ß in the intestine; LL fish showed significantly increased expressions of TNF-α, IFN-γ, lyzc, hsp70 and IL-1ß in the intestine and liver; and intestine lyzc, hsp70 and IL-1ß and liver TNF-α, IFN-γ, hsp70 and IL-1ß were significantly increased in LR+LL fish. Following a 6-week period of being fed probiotics or a control diet, the tilapia were challenged with an intraperitoneal injection of 20 µl of the pathogenic Streptococcus agalactiae (WC1535) (1 × 105 CFU/ml). The survival rates of the probiotic-fed groups were significantly higher than that of the CK group, and the LL group had the highest survival rate. High-throughput sequencing revealed a significantly higher presence of JCM5805 in the guts of LL fish during the period of probiotic application, but this was no longer detected in all LL samples 1 week post cessation of probiotic administration. Cessation of probiotic administration led to disorders of individual gut microbes within the LR and LL groups. Statistical analysis (LEfSe) demonstrated that three phyla, namely, Bacteroidetes, Fusobacteria and Actinobacteria were enriched in the CK group, while the abundance of Proteobacteria was greater in the probiotic-fed fish. At the genus level, Plesiomonas, which includes potential pathogens of fish, were significantly decreased in the probiotic-fed groups. In contrast, a significant increase of Rhizobium and Achromobacter, which can produce a variety of enzymes with cellulolytic and pectolytic activity, were observed in fish fed with probiotics, indicating that dietary probiotics were helpful in the propagation of some probiotic bacteria. Our data revealed that JCM1136 and JCM5805, as a feed additive at 108 CFU/g feed, could improve intestinal morphology, enhance immune status and disease resistance, and affect the gut microbiota of tilapia; thus, these additives could be used as probiotics for juvenile Nile tilapia. JCM5805 was more effective than JCM1136 or the mixture of the two for promoting the growth, enhancing the immune status and disease resistance of tilapia.

Molecular characterization, expression and functional analysis of NOD1, NOD2 and NLRC3 in Nile tilapia (Oreochromis niloticus).

The nucleotide-binding oligomerization domain proteins NOD1, NOD2 and NLRC3 are cytoplasmic pattern recognition receptors (PRRs) of the Nod-like receptor (NLR) family. In the present study, the Nile tilapia (Oreochromis niloticus) NOD1 (ntNOD1), NOD2 (ntNOD2) and NLRC3 (ntNLRC3) genes were cloned and characterized. The full-length ntNOD1, ntNOD2 and ntNLRC3 genes were 3924, 3886 and 4574 bp, encoding 941, 986 and 1130 amino acids, respectively. The three Nod-like receptors have a NACHT domain and a C-terminal leucine-rich repeat (LRR) domain. In addition, ntNOD1 and ntNOD2 have a N-terminal CARD domain (ntNOD2 has two). Phylogenetic analysis showed that the three NLRs are highly conserved. Tissue expression analysis of the three receptors revealed that the highest mRNA and protein levels of ntNOD1, ntNOD2 and ntNLRC3 were in the spleen. The expression patterns of NLRs during embryonic development showed that the expression levels of ntNOD2 and ntNLRC3 significantly increased from 2 to 8 days post-fertilization (dpf). The expression levels of ntNOD1 significantly increased from 2 to 6 dpf, decreased at 7 dpf and then increased at 8 dpf. Upon stimulation with an intraperitoneal injection of Streptococcus agalactiae, expression levels of the ntNOD1, ntNOD2 and ntNLRC3 mRNA and protein were clearly altered in the blood, spleen, kidney, intestine and gill. Furthermore, after cotransfection with an NF-κB reporter plasmid, NF-κB activation in ntNOD1-overexpressing 293T cells significantly increased compared with that in control cells, before or after i-EDPA-stimulation. By contrast, compared with control, ntNOD2 and ntNLRC3 had no effect on NF-κB activation in 293T cells, when their potential ligands were not stimulated. However, after MDP-stimulation, ntNOD2 and ntNLRC3 overexpression increased NF-κB activation in 293T cells. NOD1 and NLRC3 were uniformly distributed throughout the cytoplasm in 293T cells, whereas NOD2 was distributed throughout the cytoplasm and nucleus. Our results indicate that the three Nod-like receptors are functionally conserved and may play pivotal roles in defense against pathogens such as Streptococcus agalactiae.

Characterization and expression of Na+/K+-ATPase in gills and kidneys of the Teleost fish Oreochromis mossambicus, Oreochromis urolepis hornorum and their hybrids in response to salinity challenge.

Tilapia (Oreochromis mossambicus, O. urolepis hornorum, their hybrids O. mossambicus♀ × O. hornorum♂ and O. hornorum♀ × O. mossambicus♂) were exposed to a high salinity environment to evaluate their osmoregulatory responses. The plasma osmolality of all the tilapia species were elevated with the salinity challenge. The activities of Na+/K+-ATPase (NKA) in both the gill and kidney showed a similar increased change tendency compared with the control. The distribution of NKA α1 mRNA in all the examined tissues suggested that NKA α1 has a possible housekeeping role for this isoform. The amount of NKA α1 mRNA in the gill and kidney was elevated in the four fishes with similar expression patterns after transfer from freshwater to seawater. The NKAα1 mRNA expression levels in the gill reached their peak level at 24 h after transfer (P < 0.01) compared to the freshwater group, following decreases in the pretreatment level at 48 h (P > 0.05). However, the NKAα1 mRNA expression levels in the kidney were not significantly affected with increasing environmental salinity (P > 0.05). The differences in the responses to saltwater challenge may be associated with differences in saltwater tolerance between the four tilapia. The drastic increase in the plasma osmolality, NKA activities and mRNA expression suggested that the hybrids (O. mossambicus♀ × O. hornorum♂) possess heterosis in salinity responsiveness compared to that of both the parents, indicating a maternal effect on the salinity tolerance of the tilapia hybrids. This study provides a theoretical basis to further study the mechanism of fish osmoregulation response to salinity challenge.

[Identification and pathological observation of a pathogenic Plesiomonas shigelloides strain isolated from cultured tilapia (Oreochromis niloticus)].

OBJECTIVE: A mass mortality of tilapia broke out in an aquaculture farm in Panyu, Guangdong Province in May, 2013. Affected fish showed blackening of body color, haemorrhageing on surface, scales shedding, fin rotting, and the presence of yellow liver, dark red spleen, enlarged gallbladder and ascitic fluid in the abdominal cavity. The purpose of this research was isolating and identifying the pathogen. METHODS: We isolated a suspicious bacteria strain PYS1 from diseased fish with significant pathological signs. The homology of 16S rRNA gene sequence of strain PYS1 and its morphological, cultural, and physical and chemical characteristics were studied for its identification. Its pathogenicity was investigated by recursive infection experiment and histopathological study. Its effective medicines was screened by antibiotic sensitive test. RESULTS: The results showed that strain PYS1 was Plesiomonas shigelloides clustered with P. shigelloides strains isolated from other fishes in the molecular phylogenetic tree of 16S rRNA gene sequences. Strain PYS1 was multiple drug resistant and only sensitive to a small part of 31 tested antibiotics (e.g., ceftriaxone, cefaclor, cefazolin, etc.). The symptoms of tilapia (O. niloticus) artificially infected with strain PYS1 were similar with natural infected fish. The half lethal dose (LD50) of strain PYS1 to tilapia was 1.425 x 10(8) CFU per fish. Paraffin sections showed intestine, liver, spleen, kidney and heart tissue injury caused by the strain. CONCLUSION: Our study demonstrated that P. shigelloides was the pathogen of cultured tilapia in the aquaculture farm and first reported its bacterial pathogenicity on Nile tilapia.

A left-sided destroyed lung in a 11-year-old girl: A rare sequela after Mycoplasma pneumoniae infection.

Mycoplasma pneumoniae infection is one of the most common causes of community-acquired pneumonia in children, usually experiencing a favorable prognosis.Cases of M. pneumoniae infection resulting in respiratory failure, severe pulmonary or extrapulmonary sequelae, and death are relatively rare.Currently, no cases related to a destroyed lung with Mycoplasma-associated infection have been reported.Therefore, we report a case of a destroyed lung in an 11-year-old girl with Mycoplasma-associated infection.

Major histocompatibility complex class IIA and IIB genes of Nile tilapia Oreochromis niloticus: genomic structure, molecular polymorphism and expression patterns.

Major histocompatibility complex (MHC) is a large genomic region characterized by extremely high polymorphism, and it plays an important role in the immune response of vertebrates. In the present study, we isolated MHC class II genes from Nile tilapia in order to investigate the immune mechanism in tilapia and develop better strategies for disease prevention. Moreover, we cloned the full-length cDNA sequences of MHC IIA and IIB from Nile tilapia by the RACE approach. In addition, the genomic structure, molecular polymorphism and expression patterns of MHC II genes in Nile tilapia were also examined. Compared with that of other teleosts, Nile tilapia MHC class IIA contained four exons and three introns. The deduced amino acid sequence of the MHC IIA molecule shared 25.4-64.5% similarity with those of other teleosts and mammals. Six exons and five introns were identified from Nile tilapia MHC IIB, and the deduced amino acid sequence shared 26.9-74.7% similarity with those of other teleosts and mammals. All the characteristic features of MHC class II chain structure could be identified in the deduced sequences of MHC IIA and IIB molecules, including the leader peptide, α1/ß1 and α2/ß2 domains, connecting peptide and transmembrane and cytoplasmic regions, as well as conserved cysteines and N-glycosylation site. A total of 12 MHC IIA alleles were identified from six individuals. Four alleles originating from a single individual suggested that at least four MHC IIA loci existed. Moreover, 10 MHC IIB alleles were identified, among which four were detected in a single individual, suggesting that at least four MHC IIB loci existed. The expression of MHC IIA and IIB at the mRNA level in 10 types of normal tissues was determined using quantitative real-time PCR analysis. The highest expression level was detected in stomach and gill, whereas the lowest expression was detected in muscle and brain. Furthermore, MHC IIA and IIB were probably two candidate immune molecules involved in the resistance against streptococcosis, because their expression was significantly up-regulated in gill, kidney, intestine and spleen after the intraperitoneal injection of Streptococcus agalactiae.

Metabolism responses in the intestine of Oreochromis mossambicus exposed to salinity, alkalinity and salt-alkalinity stress using LC-MS/MS-based metabolomics.

Multiple abiotic stresses are imposed on fish as a result of unprecedented changes in temperature and precipitation patterns in recent decades. It is unclear how teleosts respond to severe ambient salinity, alkalinity, and saline-alkalinity in terms of their metabolic and molecular osmoregulation processes. The metabolic reactions in the intestine of Oreochromis mossambicus under salinity (25 g/L, S_C), alkalinity (4 g/L, A_C), and saline-alkalinity (salinity: 25 g/L & alkalinity: 4 g/L, SA_C) stresses were examined in this research utilizing LC-MS/MS-based metabolomics. The findings demonstrated that the three osmotic-stressed groups' metabolic profiles were considerably different from those of the control group. Osmolytes, energy sources, free amino acids, and several intermediate metabolites were all synthetically adjusted as part of the osmoregulation associated with the salinity, alkalinity, and saline-alkalinity stress. Following osmotic stress, osmoregulation-related pathways, including the mTOR signaling pathway, TCA cycle, glycolysis/gluconeogenesis, etc., were also discovered in the intestine of O. mossambicus. Overall, our findings can assist in better comprehending the molecular regulatory mechanism in euryhaline fish under various osmotic pressures and can offer a preliminary profile of osmotic regulation.

Controlled release of curcumin from gelatin hydrogels by the molecular-weight modulation of an oxidized dextran cross-linker.

Controlled drug delivery could minimize side effects while maintaining a high local dose. Herein, a hydrogel carrier was prepared by forming dynamic imine bonds between gelatin and oxidized dextran (ODex) of different molecular weights (Mw = 10, 70, and 150 kDa). The morphology, thermal stability, rheology, mechanical properties, and swelling properties of the hydrogels and the controlled release of curcumin were characterized. When dextran with a higher Mw was used, the ODex contained more aldehyde groups, which led to a higher degree of cross-linking, considerably shorter gel time, decreased hydrogel porosity, and well-controlled release of curcumin. In addition, the cross-linked hydrogels exhibited not only high thermal stability but also excellent mechanical properties. However, because the matrix was hydrophilic, the swelling properties of the hydrogels were not significantly affected by the Mw of ODex. These observations suggest an approach for designing nutrient delivery carriers with improved controlled release.

Mitochondrial DNA and Microsatellite Analyses Showed Panmixia between Temporal Samples in Endangered Anguilla japonica in the Pearl River Basin (China).

The Japanese eel (Anguilla japonica) is a commercially important species in East Asia, the abundance of which has rapidly decreased in recent decades. The fishery resource in the Pearl River basin has mainly deteriorated due to overexploitation and habitat degradation. Knowledge on its genetic status is indispensable for resource management. In this study, we explored the temporal genetic structure of A. japonica on the basis of the concatenated sequences of two mitochondrial fragments (mtDNA) and eight microsatellite markers. A total of nine temporal samples (N = 127) were collected during 2019 and 2021 from Jiangmen City, China, which is located in the Pearl River estuary. mtDNA sequence analysis showed a high level of haplotype diversity, and yielded 124 haplotypes with ranging from 9 to 19 in temporal samples. All microsatellite loci were polymorphic among each of the nine temporal samples, with 150 alleles identified across all samples. Pairwise FST values were low and nonsignificant according to both mtDNA and microsatellite markers. STRUCTURE analysis showed that all temporal samples were not clearly differentiated from each other. The yielded outcomes supported a panmictic pattern in different temporal A. japonica samples. Therefore, our results call for the management of A. japonica as a single unit and joint conservation strategy of the species, since overexploitation in any region will decrease its global resource.

Comparison of the physical stabilities and oxidation of lipids and proteins in natural and polyphenol-modified soybean protein isolate-stabilized emulsions.

Oil-in-water emulsions are widely used in the food industry; however, lipids are often easily oxidized, which may adversely affect food quality. Herein, we investigated the effects of alkali treatment, free radical induction, and carbodiimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS)-mediated synthetic methods on the structures and antioxidant properties of soy protein isolate (SPI)-gallic acid (GA) conjugates and the physical stabilities and protein-lipid co-oxidation properties of the resulting emulsions. These three methods are well established; however, their effects on the same protein-phenolic compound system have not been directly compared. Additionally, the co-oxidation of proteins and oils in emulsions remains unexplored. Alkali treatment yielded superior antioxidant properties compared to those obtained using free radicals or EDC/NHS, as this method was more likely to yield CS bonds and resulted in an increased quantity of grafted GA. Spectroscopic analysis showed that alkali treatment promoted GA oxidation and thereby increased GA-protein interactions and the quenching of tryptophan fluorescence. Correspondingly, EDC/NHS-mediated conjugation retained the activity of the hydroxyl groups of GA to the largest extent. Moreover, the grafting of GA improved the physical and oxidative stabilities of the emulsions. In particular, EDC/NHS-mediated conjugation produced an emulsion with optimal oxidative stability owing to its effective inhibition of lipid and protein oxidation. Conversely, the conjugates synthesized via alkali treatment and free radical induction displayed less inhibition of lipid oxidation and promoted protein oxidation. In conclusion, optimized protein-phenolic compound conjugates for use in developing nutritional fortification products with longer shelf lives can be obtained by using appropriate synthetic methods.

Different interactions driving the binding of soy proteins (7S/11S) and flavonoids (quercetin/rutin): Alterations in the conformational and functional properties of soy proteins.

The purpose of this research was to comparatively investigate the interactions between bioactive flavonoids (quercetin and rutin) and two predominant soy proteins (ß-conglycinin and glycinin), and the structural and functional properties of their complexes. The binding affinities of quercetin/rutin toward 7S/11S were structure-dependent, in that rutin had a higher binding affinity than that of quercetin, and 11S exhibited higher affinity toward quercetin/rutin than that of 7S. The interactions in the 7S/11S-quercetin complexes were driven by van der Waals forces and hydrogen-bonding interactions, whereas the 7S/11S-rutin complexes exhibited hydrophobic interactions. Binding to quercetin or rutin altered the secondary structures (decrease in the α-helix and random coil contents and increase in the ß-sheet content), decreased the surface hydrophobicity and thermal stability, and enhanced the antioxidant capacity of 7S and 11S. These findings provide valuable information that can facilitate the design of custom-tailored protein-flavonoid macromolecules.

Effects of different surfactants on the conjugates of soybean protein-polyphenols for the preparation of ß-carotene microcapsules.

In this study, we investigated the spray-drying microencapsulation of ß-carotene in oil co-stabilized by soy protein isolate-epigallocatechin-3-gallate conjugate (SPE) and small molecule surfactants [sodium dodecyl sulfate (SDS), hexadecyl trimethyl ammonium bromide (CTAB), and tea saponin (TS)] of different concentrations [0.1, 0.5, and 1.0% (w/v)], as a prospective approach to stabilize ß-carotene. The results show that different surfactant types and concentrations significantly affect the encapsulation efficiency, water dispersibility, microstructure, and digestion of the microcapsules. Interactions between the surfactants and the SPE at the interface were found to include both synergistic and competitive effects, and they depended on the surfactant type and concentration. Moreover, the addition of SDS and TS before spray drying significantly improved the microencapsulation performance of the microcapsules and the water dispersion behavior of the corresponding spray-dried powders. The highest encapsulation efficiency was achieved for the SPE-0.1TS-encapsulated ß-carotene microcapsules. In contrast, the addition of CTAB was not conducive to microcapsule formation, resulting in poor encapsulation efficiency, water dispersibility, thermal stability, ß-carotene retention rate, and oxidation stability. In vitro gastrointestinal digestion results revealed that the addition of CTAB promotes the release of ß-carotene and improves the bioaccessibility of ß-carotene. In contrast, except for SPE-1.0SDS, the addition of SDS and TS inhibited ß-carotene release and reduced ß-carotene bioaccessibility. This study demonstrated that this novel ß-carotene encapsulation formulation can overcome stability limitations for the development of ß-carotene supplements with a high bioaccessibility.

Emulsions co-stabilized by soy protein nanoparticles and tea saponin: Physical stability, rheological properties, oxidative stability, and lipid digestion.

Herein, the effects of the concentration (0.1%-1.0%, w/v) and addition sequence of tea saponin (TS) on the physical stability, oxidative stability, rheological properties, and in vitro digestion of the emulsions stabilized by heat-induced soy protein isolate nanoparticles (SPs) were investigated. The results revealed that the concentration and addition sequence of TS have significant impact on the microstructure, stability, rheological properties, and in vitro digestion of the emulsions. TS was shown to not only fill the interfacial gaps but also adsorb on the particle surfaces, contributing to interfacial wettability. With increasing TS concentration, interfacial tension decay is clearly observed. Further, TS endows the droplets with electrostatic repulsion and steric resistance, preventing their flocculation, coalescence, and oxidation. Finally, in vitro digestion experiments demonstrated that the presence of TS delayed the lipid digestion of the emulsions.

Identification and expression analysis of two growth hormone receptors in zanzibar tilapia (Oreochromis hornorum).

Growth hormone plays important roles in various physiological processes such as growth, metabolism, and reproduction. In this study, two cDNAs encoding growth hormone receptor (GHR) were isolated from the liver of zanzibar tilapia (Oreochromis hornornum). The two cDNAs were 2,831 and 2,044 bp in length and named GHR1 and GHR2, respectively. GHR1 and GHR2 shared 57.4% similarity in nucleotide sequences and 33.5% similarity in deduced amino acid sequences. Consequently, it was presumed that they were two different genes. Conserved regions of GHR1 and GHR2 in zanzibar tilapia were different from those of other vertebrates. For example, conserved box2 regions of GHR1 and GHR2 in zanzibar tilapia were, respectively, WVELM and WVEFT, while it was WVEFI for GHRs in other vertebrates. Similar to other fish species, GHR1 and GHR2 were expressed in brain, gill, liver, muscle, spleen, gonad, stomach, kidney, and pituitary in zanzibar tilapia. The expression levels were the highest in liver. Unlike fathead minnow (Pimephales promelas) and mossambique tilapia (O. mossambicus), the expression levels of GHR1 in most female fish tissues were higher than those in male fish. No significant difference in GHR2 expression was found in all the tissues in male and female of zanzibar tilapia. Under fasting condition, the expressions of GHRs and IGF-II were significantly up-regulated (P < 0.05) in liver, while the expression of IGF-I remained stable. This observation would contribute to understanding the evolution of the GHR family in further investigation of growth regulation of zanzibar tilapia.

[Dynamic expression of CyclinD1 and p21CIP1 during lung development in rats].

OBJECTIVE: CyclinD1 and p21CIP1 are major proteins to regulate lung cell proliferation and involved in lung development and lung injury reparation. This study aimed to explore the expression manners of CyclinD1 and p21CIP1 at canalicular, saccular and alveolar stages during lung development in Sprague-Dawley rats. METHODS: Lung tissues were obtained from fetal rats of 20 and 21 days gestational ages, and neonatal rats at 0, 3, 7, 14 and 21 days (n=6). Lung tissues were used for histopathology and the protein analysis of CyclinD1 and p21CIP1 (immunohistochemistry and Western blot). RESULTS: The strongest expression of CyclinD1 and the weakest expression of p21CIP1 occurred at 20-21 days gestation (canalicular stage). At the canalicular stage, CyclinD1 was mainly expressed in epithelial cells, and the expression of p21CIP1was negative. At the saccular stage, the expression of CyclinD1 decreased significantly and the p21CIP1 expression increased significantly. Positive expression of CyclinD1 and p21CIP1 was found in epithelial cells and interstitial cells. At the alveolar stage, the CyclinD1 expression was the lowest and the p21CIP1 expression was the highest. The positive expression of CyclinD1 was found in interstitial cells and that of p21CIP1 was found in epithelial cells. CONCLUSIONS: The location and quantity of CyclinD1 and p21CIP1 expression are different at various stages during lung development in rats. A strongest CyclinD1 expression found in the canalicular stage may be associated a high lung cell proliferation. A strongest p21CIP1 expression found in the alveolar stage may be associated with alveolar maturity.