母乳联合鼠李糖乳杆菌预防早产儿喂养不耐受的疗效观察

目的 观察母乳联合鼠李糖乳杆菌预防早产儿喂养不耐受的临床疗效。方法 选取2016年2月至2017年2月武汉大学人民医院收治的80例生后24 h内入住本院儿科新生儿病房的早产儿,随机分配成2组,其中研究组40例,对照组40例。研究组以新鲜母乳联合鼠李糖乳杆菌早期开奶喂养,对照组以早产儿配方奶早期开奶喂养,比较2组早产儿喂养不耐受症的发生率及其恢复出生体重时间、达全胃肠道喂养时间、体重增长速度和住院时间的差异。结果 研究组患儿喂养不耐受发生率低于对照组,研究组恢复出生体重时间、达全胃肠道喂养时间及住院时间均短于对照组（P<0.05）,研究组患儿体重增加速度快于对照组（P<0.05）。结论 早产儿早期喂养母乳联合鼠李糖乳杆菌能够较早建立全胃肠道营养,减少早产儿喂养不耐受症的发生率,促进患儿体重增长,缩短恢复出生体重时间及达全胃肠道喂养时间,降低住院天数。     

A novel leptin antagonist peptide inhibits breast cancer growth in vitro and in vivo

The role of the obesity cytokine leptin in breast cancer progression has raised interest in interfering with leptin's actions as a valuable therapeutic strategy. Leptin interacts with its receptor through three different binding sites: I–III. Site I is crucial for the formation of an active leptin–leptin receptor complex and in its subsequent activation. Amino acids 39‐42 (Leu‐Asp‐Phe‐Ile‐ LDFI) were shown to contribute to leptin binding site I and their mutations in alanine resulted in muteins acting as typical antagonists. We synthesized a small peptide based on the wild‐type sequence of leptin binding site I (LDFI) and evaluated its efficacy in antagonizing leptin actions in breast cancer using in vitro and in vivo experimental models. The peptide LDFI abolished the leptin‐induced anchorage‐dependent and ‐independent growth as well as the migration of ERα‐positive (MCF‐7) and ‐negative (SKBR3) breast cancer cells. These results were well correlated with a reduction in the phosphorylation levels of leptin downstream effectors, as JAK2/STAT3/AKT/MAPK. Importantly, the peptide LDFI reversed the leptin‐mediated up‐regulation of its gene expression, as an additional mechanism able to enhance the peptide antagonistic activity. The described effects were specific for leptin signalling, since the developed peptide was not able to antagonize the other growth factors' actions on signalling activation, proliferation and migration. Finally, we showed that the LDFI pegylated peptide markedly reduced breast tumour growth in xenograft models. The unmodified peptide LDFI acting as a full leptin antagonist could become an attractive option for breast cancer treatment, especially in obese women.     

味觉刺激后瘦素及其受体表达的变化

目的 :研究经口腔味觉刺激后 ,大鼠血清瘦素水平和脑瘦素受体表达情况。方法 :给大鼠口腔味觉刺激 ,味觉刺激物包括 3mol/L蔗糖 ,5mmol/L糖精钠 ,0 .1mol/LNaCl,0 .0 1mol/LHCl,1mmol/L奎宁H2 SO4和 0 .1mol/L谷氨酸钠 ,采用大鼠瘦素放免试剂盒测定血清瘦素水平。应用免疫组织化学ABC法对大脑切片进行染色 ,一抗为特异性羊抗瘦素受体IgG。 结果 :与对照组 (以蒸馏水代替味觉刺激物 )相比 ,仅甜味组 (包括蔗糖和糖精钠 )血清瘦素水平升高 (P <0 .0 5 )。杏仁核、下丘脑、臂旁核和孤束核等与味觉和摄食明显相关的核团均存在瘦素受体免疫反应 (LR IR)阳性细胞 ,但是阳性细胞数目在味觉刺激组和对照组间无显著性差异。结论 :给大鼠甜味觉刺激后 ,血清瘦素水平升高。并且杏仁核这个在摄食的发动和引导中起重要作用的核团存在LR IR阳性细胞。这些结果提示瘦素可能通过调节味觉感受而影响摄食 ,有必要对瘦素在味觉感受方面的作用进一步研究     

Recent insights into body weight control: from physiology to pathology.

Over the past several years, new modulators of feeding and body weight have been discovered, and our knowledge of the mechanisms and neurohumoral interactions between anorexigenic and orexigenic compounds has increased dramatically. This review aims to summarize the present knowledge of the role of leptin and several hypothalamic neuropeptides, such as neuropeptide Y (NPY), corticotropin-releasing hormone (CRH) and melanocortins, in the regulation of appetite and body weight. It also presents the progress made in the design of interactions between leptin and hypothalamic peptides in the regulation of feeding. The role of these compounds in the pathogenesis of obesity in animals and humans, and their potential usefulness in the treatment of this disorder, are discussed.     

Effect of leptin gene polymorphism on the breeding value of milk production traits in Iranian Holstein

New molecular techniques focused on genome analysis, open new possibilities for more accurate evaluation of economiclly important traits in farm animals. Milk production traits are typical quantitative characteristics controlled by a number of genes. Mutations in their sequences may alter animal performance as well as their breeding values. In this study, we investigated the effect of Kpn2I restriction fragment length polymorphisms in the leptin gene, on bull breeding values for milk yield, fat, and protein yield, and their percentage. In order to test for an association between the leptin single-nucleotide polymorphism in exon 2 and milk productivity, we genotyped 134 Iranian Holstein bulls. Breeding values for milk-related traits (milk yield, fat, and protein yield and percentage) were estimated using the BLUP based on an animal model. The effect of the genotypes of Kpn2I polymorphism on the breeding values for milk-related traits was examined using least square methods. The T allele frequency was 0.425. Genotypes were distributed according to the Hardy-Weinberg equilibrium. Bulls with TT genotype had higher milk, fat and protein yield compared with TC and CC bulls (P < 0.05). Bulls with CC genotype had higher protein percentage compared with TT and TC bulls (P < 0.05). The association between leptin polymorphism with milk production traits suggests that this marker may be useful for selection based on molecular information.     

成都地区母乳双歧杆菌对婴儿肠道双歧杆菌构建的影响

观察不同喂养方式婴儿肠道双歧杆菌的构建规律,初步探索母乳双歧杆菌与婴儿肠道双歧杆菌之间的相关性。

在四川大学华西第二医院纳入57对母婴志愿者,根据喂养方式分为母乳（BF）组（n = 31）和混合喂养（MF）组（1月龄内添加奶粉时间＞8 d,n = 26）,收集产妇产后0天、第30天的母乳,以及婴儿出生后0天、第7天、第30天的粪便。采用实时荧光定量PCR测定母乳和婴儿粪便中双歧杆菌属、角双歧杆菌（B. angulatum）、链双歧杆菌（B. catnulatum）、齿双歧杆菌（B. dentium）、短双歧杆菌（B. breve）、两歧双歧杆菌（B. bifidum）、长双歧杆菌长亚种（B. longum subsp. longum）、长双歧杆菌婴儿亚种（B. longum subsp. infantis）及青春双歧杆菌（B. adolescentis）的丰度。观察2组婴儿肠道双歧杆菌0―7―30天的构建规律;分析母乳双歧杆菌与婴儿肠道双歧杆菌之间的相关性。

0―7―30天2组婴儿肠道双歧杆菌构建过程存在差异。母乳双歧杆菌与婴儿肠道双歧杆菌存在相关：0 d时,BF组母乳B. angulatum与婴儿肠道B. longum subsp. longum呈正相关（r = 0.962 3,P = 0.037 7）,母乳B. longum subsp. infantis与婴儿肠道B. longum subsp. longum、B. breve及B. dentium呈正相关（r = 0.998 4,P = 0.036 1;r = 1.000 0,P = 0.000 2;r = 1.000 0,P = 0.002 3）,母乳B. catnulatum与婴儿肠道B. dentium呈正相关（r = 0.997 7,P = 0.043 1）;30 d时,BF组母乳B. adolescentis与婴儿肠道双歧杆菌属呈正相关（r = 0.532 2,P = 0.013 0）,母乳B. longum subsp. longum与婴儿肠道B. breve、B. catnulatum呈负相关（r = ‒0.458 9,P = 0.048 1;r = ‒0.520 7,P = 0.032 1）,母乳B. catnulatum与婴儿肠道B. angulatum呈负相关（r = ‒0.682 8,P = 0.007 1）,母乳双歧杆菌属与婴儿肠道B. longum subsp. longum呈负相关（r = ‒0.714 3,P = 0.046 5）;30 d时,MF组母乳B. bifidum与婴儿肠道B. longum subsp. longum呈正相关（r = 0.579 2,P = 0.030 0）,母乳B. angulatum与婴儿肠道B. catnulatum呈正相关（r = 0.550 1,P = 0.022 1）。

喂养方式会影响婴儿肠道双歧杆菌构建;母乳中存在双歧杆菌且与婴儿肠道双歧杆菌相关,母乳喂养与婴儿肠道双歧杆菌的相关性更密切,尤其0 d时相关性更强;混合喂养可能由于奶粉的添加使母乳双歧杆菌与婴儿肠道双歧杆菌的相关性降低,规律性减弱。

     

Antimicrobial potential of four Lactobacillus strains isolated from breast milk

AIMS: The antimicrobial potential of four lactobacilli (Lactobacillus salivarius CECT5713, Lactobacillus gasseri CECT5714, L. gasseri CECT5715 and Lactobacillus fermentum CECT5716), isolated from fresh human breast milk, was evaluated in this study and compared with Lactobacillus coryniformis CECT5711, a reuterin-producing strain isolated from an artisan goat's cheese. METHODS AND RESULTS: Agar diffusion tests, competitive adhesion assays and mucin expression assays were carried out in order to value the antibacterial properties of the lactobacilli strains. The antibacterial capability of the strains was tested in vivo by using a murine infection model with Salmonella choleraesuis. The results revealed that all the strains studied, displayed antibacterial properties against pathogenic bacteria. However, the antibacterial potential varied among the lactobacilli tested and, in fact, L. salivarius CECT5713 showed not only the best in vitro antibacterial activity, but also the highest protective effect against a Salmonella strain in the murine infection model. CONCLUSION: The four breast-milk lactobacilli, and particularly L. salivarius CECT5713, possess potent antibacterial activities that result in a higher protection against S. choleraesuis CECT4155 in a mouse infection model. SIGNIFICANCE AND IMPACT OF THE STUDY: These results suggest that lactobacilli from breast milk could contribute to an anti-infective protection in neonates and would be excellent candidates for the development of infant probiotic products.     

Role of leptin in the regulation of food intake in fasted mice

Leptin is well acknowledged as an anorexigenic hormone that plays an important role in feeding control. Hypothalamic GABA system plays a significant role in leptin regulation on feeding and metabolism control. However, the pharmacological relationship of leptin and GABA receptor is still obscure. Therefore, we investigated the effect of leptin or combined with baclofen on the food intake in fasted mice. We detected the changes in hypothalamic c‐Fos expression, hypothalamic TH, POMC and GAD67 expression, plasma insulin, POMC and GABA levels to demonstrate the mechanisms. We found that leptin inhibit fasting‐induced increased food intake and activated hypothalamic neurons. The inhibitory effect on food intake induced by leptin in fasted mice can be reversed by pretreatment with baclofen. Baclofen reversed leptin's inhibition on c‐Fos expression of PAMM in fasted mice. Therefore, these results indicate that leptin might inhibit fasting‐triggered activation of PVN neurons via presynaptic GABA synaptic functions which might be partially blocked by pharmacological activating GABA‐B. Our findings identify the role of leptin in the regulation of food intake.     

Characterization of gut microbiota and short-chain fatty acid in breastfed infants with or without breast milk jaundice

This study aims to investigate the gut microbiota and metabolites in breastfed infants with breast milk jaundice (BMJ) using gut microbiome–metabolomics. Breastfed newborns diagnosed with BMJ and those without BMJ (control group) were enrolled. Faecal samples were collected from the participants and subjected to high-throughput sequencing of the 16s rDNA V3 and V4 regions of the gut flora and metabolomics of short-chain fatty acids (SCFAs). Proteobacteria, Fimicutes and Actinobacteria were the main bacteria at the phylum level. Eshcerichia-Shigella and Enterobacteriacea were the main bacteria at the genus level. The difference between the two groups was compared. Compared to the control group, the amount of Streptococcus was significantly increased while the amount of Enterococcus was significantly decreased in the faeces from infants with BMJ. Functional prediction analysis of 16S found that biosynthesis of penicillin and cephalosporin significantly increased in the BMJ group. Gas chromatography–mass spectrometry detection of SCFAs revealed that levels of acetic acid and propionic acid were significantly lower in the BMJ group than in the control group. The reduced levels of acetic acid and propionic acid may be related to the increase in Streptococcus and decrease in Enterococcus, both of which may contribute to BMJ.     

Immune-related microRNAs are abundant in breast milk exosomes

Breast milk is a complex liquid rich in immunological components that affect the development of the infant's immune system. Exosomes are membranous vesicles of endocytic origin that are found in various body fluids and that can mediate intercellular communication. MicroRNAs (miRNAs), a well-defined group of non-coding small RNAs, are packaged inside exosomes in human breast milk. Here, we identified 602 unique miRNAs originating from 452 miRNA precursors (pre-miRNAs) in human breast milk exosomes using deep sequencing technology. We found that, out of 87 well-characterized immune-related pre-miRNAs, 59 (67.82%) are presented and enriched in breast milk exosomes (P < 10(-16), χ(2) test). In addition, compared with exogenous synthetic miRNAs, these endogenous immune-related miRNAs are more resistant to relatively harsh conditions. It is, therefore, tempting to speculate that these exosomal miRNAs are transferred from the mother's milk to the infant via the digestive tract, and that they play a critical role in the development of the infant immune system.     

Human milk nutrient fortifiers alter the developing gastrointestinal microbiota of very-low-birth-weight infants

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母乳联合妈咪爱对极低出生体重儿炎性细胞因子的影响

目的 探讨母乳联合妈咪爱对极低出生体重儿（ELBWI）炎性细胞因子的影响。方法 采用前瞻性研究法,选择2012年2月至2016年5月在我院诊治的ELBWI 76例,根据随机数字表法分为观察组与对照组各38例,对照组给予母乳喂养,观察组给予母乳联合妈咪爱喂养,两组均给予暖箱保暖、治疗原发病等对症治疗。治疗14 d后,观察两组患儿的预后与炎性细胞因子表达情况。结果 观察组与对照组的喂养不耐受率分别为7.9%和26.3%,观察组明显少于对照组（χ2=3.289,P0.05）。观察组治疗后的血清IL-6与TNF-α含量分别为（16.33±11.61）ng/L和（25.23±8.92）ng/L,均明显低于对照组的（26.35±11.56）ng/L与（33.72±10.36）ng/L（P<0.05）,且都明显低于治疗前（P<0.05）。结论 母乳联合妈咪爱喂养可改善ELBWI早期喂养的耐受性,提高患儿免疫功能,降低炎性细胞因子IL-6与TNF-α的表达,从而对生长发育有一定促进作用。     

Identification and isolation from breast milk of ceruloplasmin-lactoferrin complex

The presence of a complex of the copper-containing protein ceruloplasmin (Cp) with lactoferrin (Lf) in breast milk (BM) is shown for the first time. In SDS-free polyacrylamide gel electrophoresis (PAGE), electrophoretic mobility of Cp in BM is lower than that of plasma Cp, coinciding with the mobility of the complex obtained upon mixing purified Cp and Lf. Affinity chromatography of delipidated BM on Cp-Sepharose resulted in retention of Lf. SDS-PAGE of the 0.3 M NaCl eluate revealed a single band with M_r ∼ 78,000 that has the N-terminal amino acid sequence of Lf and reacts with antibodies to that protein. Synthetic peptides R-R-R-R (the N-terminal amino acid stretch 2–5 in Lf) and K-R-Y-K-Q-R-V-K-N-K (the C-terminal stretch 29–38 in PACAP 38) caused efficient elution of Lf from Cp-Sepharose. Cp-Lf complex from delipidated BM is not retained on the resins used for isolation of Cp (AE-agarose) and of Lf (CM-Sephadex). Anionic peptides from Cp-(586–597), (721–734), and (905–914)—provide an efficient elution of Cp from AE-agarose, but do not cause dissociation of Cp-Lf complex. When anti-Lf is added to BM flowed through CM-Sephadex, Cp co-precipitates with Lf. Cp-Lf complex can be isolated from BM by chromatography on CM-Sephadex, ethanol precipitation, and affinity chromatography on AE-agarose, yielding 98% pure complex. The resulting complex Cp-Lf(1: 1) was separated into components by chromatography on heparin-Sepharose. Limited tryptic hydrolysis of Cp obtained from BM and from blood plasma revealed identical proteolytic fragments. Published in Russian in Biokhimiya, 2006, Vol. 71, No. 2, pp. 208–215.     

Heavy metal concentrations in the breast milk of Saudi women

Lead, cadmium, and mercury concentrations were determined in breast milk of Saudi lactating mothers from Riyadh and Al-Ehssa regions in Saudi Arabia who were not occupationally exposed. The mean levels for cadmium, lead, and mercury were 1.732 μg/L, 31.671 μg/L, and 3.100 μg/L, respectively. In contrast to mercury, mothers living in the Al-Ehssa region had significantly higher cadmium and lead concentrations in their breast milk than those in the Riyadh region. The estimated weekly intakes of cadmium, lead, and mercury of breast-fed infants in this study were in some cases higher than the Provisional Tolerance Weekly Intake (PTWI) recommended by FAO/WHO, which pose a threat to their health. This necessitates the urgent need to undertake a comprehensive study to determine the sources of exposure to these heavy metals. Breast-feeding is of great benefical value for the infant’s development; therefore, efforts should be made to prevent its contamination with environmental pollutants.     

Peptidomic analysis reveals multiple protection of human breast milk on infants during different stages

It has been shown that human breast milk (HBM) is an important nutrient for the growth and development of newborns. Currently, peptide drugs provide promising regimes in neonatal disease treatment, especially peptides from HBM that exhibit multiple functions within cells. To explore the potential biological function peptides among the colostrum, transition and mature milk from mother of extremely low birth weight children (the samples were collected from Women's Hospital of Nanjing Medical University from December 2016 to February 2017). A total of 3,182 nonredundant peptides were identified and compared among colostrum, transitional and mature milk using liquid chromatography/mass spectrometry technology, and the numbers and fragments of peptides were various. The isoelectric point and molecular weight analysis of the differentially expressed peptides basically accord with the range of mass spectrometry identification (<3 kDa). Gene Ontology analysis and Pathway analysis, restriction sites analysis, as well as bioinformatics analysis showed that these differentially expressed peptides enriched a variety of biological processes. We identified several putative peptides that might have bioactive effects in diseases and development of newborns, which will inform further functional investigations. Our preliminary research provided a better understanding of the function of peptides during the newborn periods. Furthermore, it laid a foundation for discovering new peptide drugs in neonatal disease treatment.     

穴位艾灸治疗产后乳汁不足的疗效观察

目的观察穴位艾灸治疗产后乳汁不足的效果。方法选择2016年3月~2017年3月在我院产科分娩后乳汁不足产妇100例,随机分为对照组和治疗组各50例。对照组采用产科产后乳汁不足的常规护理,治疗组在对照组的基础上加艾灸膻中穴、乳根穴和少泽穴治疗,并对两组产妇乳汁分泌情况和婴儿的哺乳情况进行比较。结果治疗组乳汁分泌正常有效率为90%,对照组乳汁分泌正常有效率为60%,两组比较差异有显著统计学意义(P0.05)。结论艾灸膻中穴、乳根穴和少泽穴治疗产后乳汁不足的疗效较好,可明显改善产后母乳不足,是治疗产后乳汁不足的有效措施。