This consensus aims to standardize the diagnosis and management of neonatal lactose intolerance (LI). A multidisciplinary working group was established under the auspices of the Preterm Birth International Collaborative (Australasia Branch). The development process included a comprehensive literature review of databases, which identified 61 publications. Recommendations were formulated using the Grading of Recommendations, Assessment, Development and Evaluation system for evidence evaluation. A modified Delphi method was used to achieve consensus. The consensus establishes that neonatal LI presents with nonspecific gastrointestinal symptoms, requiring stool assessment for diagnosis. Screening tests include fecal reducing sugar or urinary galactose tests, and the diagnosis is confirmed through elimination testing while excluding mimics. Treatment prioritizes lactase-supplemented breastfeeding; formula-fed infants receive lactase or reduced-lactose formulas. Probiotics with β-galactosidase activity can be used as an adjuvant therapy, and evidence-based lactase preparations are recommended, particularly for preterm infants. Treatment duration should be individualized, with a general minimum of 2 weeks; however, the routine use of lactose-free formula to prevent LI in preterm infants is not recommended. This consensus provides practical evidence-based guidance to standardize the diagnosis and management of neonatal LI, aiming to reduce variation in practice and improve patient outcomes. It emphasizes the importance of maintaining breastfeeding while incorporating lactase supplementation and highlights the need for further high-quality research, particularly randomized controlled trials, to refine diagnostic criteria and therapeutic strategies for preterm and very preterm infants. The recommendations are intended to support clinicians in making informed decisions while promoting family-centered care. Trial Registration:International Practice Guidelines Registry Platform (PREPARE-2024CN606).

Early-life gut microbial composition and fecal metabolic profiles may be altered in infants born small for gestational age (SGA), but evidence specifically concerning term SGA infants remains limited. To characterize gut microbial composition, predicted microbial functions, and fecal metabolite profiles in term SGA infants and explore their associations with early growth. This prospective nested case–control study included 62 term infants (31 SGA and 31 appropriate for gestational age [AGA]). Fecal samples were collected at 3 months. Untargeted metabolomics was performed in the full cohort, and shotgun metagenomic sequencing was conducted in a nested subset of 20 infants (10 SGA and 10 AGA). Differential taxa, KEGG orthologs (KOs), metabolites, and cross-omics correlations were analyzed. Alpha diversity did not differ significantly between groups, whereas microbial composition and species-level abundance patterns differed between SGA and AGA infants. Functional analysis identified 135 differential KOs, including 36 with higher and 99 with lower abundance in the SGA group. These KOs were mapped to 25 metabolic pathways, including galactose, butanoate, D-amino acid, aromatic amino acid, fructose–mannose, pyruvate, propanoate, and lipid metabolism. Twelve fecal metabolite features differed between groups, including 5′-methylthioadenosine, taurocholate, β-alanine, and 17α-hydroxypregnanolone. Exploratory integration identified connections among microbial species, KOs, and metabolites. Several microbial and metabolite features showed nominal associations with 6-month growth indices, but most cross-omics and metabolite–growth associations did not remain significant after false discovery rate correction. Term SGA infants showed differences in gut microbial composition, predicted metabolic potential, and fecal metabolite profiles at 3 months. Associations with subsequent growth were exploratory and require validation in larger longitudinal studies.

Objective To examine the association between caffeine citrate administration and necrotising enterocolitis (NEC) in very preterm infants (VPIs).Design A multicentre retrospective cohort study.Setting Level III neonatal intensive care units participating in the Chinese Neonatal Network.Participants The participants comprised neonates with a gestational age of under 32 weeks.Interventions Exposure to caffeine citrate administration after birth.Primary and secondary outcome measures Primary outcome measures were the incidence of NEC (≥stage IIA) and surgical NEC. Secondary outcome measures included severe neonatal morbidities, including severe intraventricular haemorrhages, severe retinopathy of prematurity, late-onset sepsis, bronchopulmonary dysplasia, death, duration of parenteral nutrition and length of neonatal intensive care unit stay.Results A total of 45 624 VPIs were included. Among 36 514 who received at least one dose of caffeine citrate, 2315 (6.3%) developed NEC (≥stage IIA) and 849 (2.3%) experienced surgical NEC. Among 9110 VPIs without caffeine exposure, 544 (6.0%) developed NEC (≥stage IIA) and 263 (2.9%) experienced surgical NEC. Caffeine citrate exposure did not affect the incidence of NEC (≥stage IIA) between groups (adjusted OR (aOR), 1.01; 95% CI 0.83 to 1.24; adjusted absolute risk (aAR), −0.15; 95% CI −1.21 to 0.92); however, its early administration (within 72 hours after birth) was associated with a lower incidence of surgical NEC (aOR, 0.76; 95% CI 0.64 to 0.89; aAR, −0.92; 95% CI −1.55 to −0.29). In subgroup analysis, caffeine citrate administration was associated with a reduction in the incidence of NEC (≥stage IIA) among VPIs receiving invasive ventilation at admission (aOR, 0.8; 95% CI 0.67 to 0.94; aAR, −2.04; 95% CI −3.82 to −0.25) and vasopressors before NEC occurred (aOR, 0.64; 95% CI 0.52 to 0.78; aAR, −5.17; 95% CI −7.94 to −2.39).Conclusions While the incidence of NEC (≥stage IIA) in VPIs has not been affected by caffeine citrate, early administration (within 72 hours after birth) was associated with a reduced incidence of surgical NEC. Furthermore, among VPIs who received invasive ventilation at admission and vasopressors, the administration of caffeine citrate was potentially correlated with a decreased occurrence of NEC (≥stage IIA).

Necrotizing enterocolitis (NEC) remains a leading cause of morbidity and mortality in preterm infants, and effective pharmacologic options are scarce. Studies have demonstrated that human milk peptides can improve NEC, β-casein 65 is a novel peptide derived from preterm human milk, yet its specific function and mechanism remain unclear. Here we evaluated the therapeutic potential role of β-casein 65 in a mouse model of NEC. β-casein 65 improved survival and mitigated abdominal distension and hematochezia in experimental NEC models. Concomitantly, β-casein 65 significantly reduced pro-inflammatory cytokines (TNF-α and IL-6 by approximately 56% and 71%, respectively) and increased intestinal barrier proteins compared with the NEC group. Using 16S rRNA sequencing and untargeted metabolomics, β-casein 65 altered gut microbiota profiling by lowering pathogenic Proteobacteria and promoting beneficial bacteria like Bifidobacterium and Akkermansia. Metabolomics analysis showed notable changes in immune-regulating metabolites, especially enriched in the IgA immune network. This finding not only provides a mechanistic understanding but also highlights β-casein 65 as a promising candidate for the development of preventative therapeutics for this devastating neonatal disease.

OBJECTIVES:To evaluate the clinical utility and translational potential of a remote jaundice monitoring system for home-based screening of neonatal hyperbilirubinemia. METHODS:A prospective self-controlled study was conducted, enrolling 538 newborns with gestational age ≥35 weeks, birth weight ≥2 000 g, and postnatal age ≤14 days at the Women's Hospital of Nanjing Medical University from March to October 2023. Four screening protocols with different predictive indicators were developed based on the Chinese Neonatal Transcutaneous Hourly Bilirubin Nomogram. The effectiveness of the system was evaluated, and the feasibility of using the remote jaundice monitoring system in actual home settings was analyzed. RESULTS:A total of 538 paired transcutaneous bilirubin (TcB) and total serum bilirubin (TSB) measurements showed a strong correlation (r=0.85, P<0.001), with 95.0% (511/538) of samples within the 95% limits of agreement. Using TcB ≥ the 95th percentile as the predictive indicator, the system achieved 100% sensitivity, 46.2% specificity, and an area under the receiver operating characteristic curve of 0.731 (95%CI: 0.682-0.780). This approach could reduce unnecessary hospital visits by 41.4% (221/538). CONCLUSIONS:The system integrates the QBH-801 transcutaneous bilirubinometer, intelligent early warning, and remote guidance services, establishing a closed-loop "hospital-to-home" management model. It demonstrates high safety and feasibility, with significant clinical translational value.

BackgroundBioactive compounds, such as human milk oligosaccharides (HMOs), impact the development of the intestinal microbiome and immune maturation in early life. They have been shown to result in positive benefits, including improved gut health, reduced frequency of infections, and age-appropriate growth when added to infant formula. However, data supporting the added value of including these HMOs in early-stage infant formula is currently lacking among Chinese infants. ObjectiveIn this double-blind randomized controlled trial including a nonrandomized reference breastfed group, we will test the efficacy of ready-to-feed infant formula containing 2 HMOs (2-fucosyllactose and lacto-N-neotetraose) on Bifidobacteria abundance, gut microbiome, gut and immune health, growth, and quality of life. MethodsHealthy infants will be enrolled between 3 and 14 days after birth at 5 sites in China and randomized to either the experimental group (fed ready-to-feed infant formula containing 2 HMOs) or the control group (fed the same ready-to-feed infant formula without the 2 HMOs), using a dynamic allocation algorithm with double blinding. Infants will receive trial formula until age 6 months and will be followed up until age 12 months. The breastfed group will serve as a reference. The primary end point will be Bifidobacteria abundance in fecal samples at 3 months, measured via quantitative polymerase chain reaction. Secondary end points will include fecal microbiome (including taxonomy, diversity, functionality, and metabolites), fecal markers of immune health, gastrointestinal tolerance, stooling patterns, immune competence (overall state of the immune system), sleep quality, growth, quality of life, medication use, and physician-reported adverse events. A 2-sided test at the 5% significance level will be used for statistical testing. ResultsThe study received ethical approval in March 2024 and will be completed by the end of 2026, which will be followed by a publication in a peer-reviewed journal. ConclusionsThe Starter Liquid Infant Formula Trial (STARLIT) will be one of the first to assess the efficacy of these 2 HMOs among Chinese infants on gut and immune health, in addition to clinically relevant outcomes such as quality of life, growth, and adverse events. This study should help to demonstrate that an increase in the growth of beneficial Bifidobacteria in response to intake of 2-fucosyllactose and lacto-N-neotetraose may have a broader impact on overall gut microbiome composition and infant gut and immune health. Trial RegistrationClinicalTrials.gov NCT06361719; https://clinicaltrials.gov/study/NCT06361719 International Registered Report Identifier (IRRID)PRR1-10.2196/66489

Necrotizing enterocolitis (NEC) is one of the most common intestinal emergencies in newborn infants. It is closely related to abnormal activation of intestinal inflammation and impaired barrier function. Studies have shown that breast milk exosomes can effectively improve NEC, but the specific mechanism remains unclear. Human breast milk exosomal cargoes are emerging as new modulators for NEC. To evaluate the function and potential mechanisms of action of peptide MEANP1 both in vivo and in vitro. MEANP1 was found to improve the survival rate, abdominal distension and hematochezia in NEC models. Intestinal histological damage was improved, along with decreased expression of pro-inflammatory factors and increased expression of intestinal barrier protein. In IEC-6 cells, MEANP1 can improve LPS-induced inhibition of proliferation, migration and barrier damage. The pro-inflammatory factors TLR4, TNF-α and IL-1β were inhibited by MEANP1. We verified by RNA-seq technique and rescue experiment that it can inhibit the phosphorylation of NF-κB by inhibiting the phosphorylation of IκBα to protect from NEC. MEANP1 is expected to be a treatment for NEC.

Objective:To compare the prevention practices of necrotizing enterocolitis (NEC) across 17 neonatal intensive care units (NICUs) in China. Methods:A web-based survey was sent to 17 level 3 NICUs in China on September 21, 2023, to evaluate the prevention strategies for NEC. Results:All 17 Neonatal Intensive Care Units (NICUs) responded to the survey. There was significant variation in the initial empirical use of antibiotics for early-onset sepsis, late-onset sepsis, and NEC among different NICUs. Out of the 17 NICUs, only 5 (29.4%) used donor human milk. Additionally, 15 (88.2%) NICUs performed routine echocardiography (Echo) in preterm infants after birth to evaluate cardiac function and/or Patent Ductus Arteriosus (PDA) status. Out of those 15 NICUs, 11 (73.3%) performed Echo within 24 to 72 hours after birth. Furthermore, 8 NICUs (47.1%) did not alter enteral nutrition management during drug treatment for PDA, while 12 NICUs (70.6%) stopped 1 or 2 feeds during red blood cell transfusion. Conclusion:The findings of this survey conducted through questionnaires revealed both differences and similarities in the strategies employed to prevent NEC in 17 NICUs in China.

Objective: To provide an updated review of the clinical profile and outcomes of delivery room chest compression (DR-CC) in China. Method: Retrospective analysis of prospectively collected data from 23 neonatal intensive care units in Jiangsu, China (2019-2021). Antepartum, delivery room, and postpartum variables in DR-CC-receiving and no-DR-CC groups were compared using uni- and multivariate analyses. The main outcome measure was survival without major morbidities at discharge. Results: Among 2120 preterm infants of <32 weeks gestational age, 112 (5.39%) received DR-CCs. Forty-two (37.50%) DR-CC-group infants survived without major morbidities at discharge, compared with 1299 (66.17%) no-DR-CC-group infants. The DR-CC group had a lower adjusted odds ratio (AOR) of survival without major morbidities (0.53 [0.31, 0.89]). In secondary outcomes, infants who received DR-CCs had more in-hospital mortality (AOR:1.95[1.12, 3.40]) and a significant increase in the rate of grade 3/4 intraventricular hemorrhage / periventricular leukomalacia or death (AOR: 2.35[1.40, 3.95]), moderate bronchopulmonary dysplasia or death (AOR: 2.02[1.21, 3.37]), >= stage 3 retinopathy of prematurity or death (AOR: 2.22[1.33, 3.69]), stage 2 necrotizing enterocolitis or death (AOR: 1.83[1.09, 3.07]) and late-onset sepsis or death (AOR:1.66[1.02, 2.70]). In DR-CC-group infants, use of a T-piece resuscitator, noninvasive respiratory support, and higher gestational age significantly influenced survival without morbidities. Conclusion: This multicenter cohort study revealed the clinical landscape of preterm infants (<32 weeks gestational age) receiving DR-CCs, showing lower survival rates without major morbidities compared to those not receiving DR-CCs.

OBJECTIVES:To explore the impact of different treatment attitudes on the survival status of extremely preterm infants (EPIs) and evaluate the mortality and occurrence of severe complications in actively treated infants, as well as their risk factors. METHODS:A retrospective analysis was conducted on perinatal data of EPIs born between January 1, 2016, and December 31, 2023, who were admitted to the neonatal intensive care unit of Nanjing Women and Children's Healthcare Hospital within 24 hours after birth. The analysis focused on the attributable risk of mortality associated with different treatment attitudes in EPIs of varying gestational ages and birth weights. A multivariate logistic regression model was used to analyze the risk factors for mortality and severe complications in the actively treated group. RESULTS:A total of 485 EPIs were included. As gestational age or birth weight increased, the attributable risk of mortality with care withdrawal increased. Active treatment significantly improved the survival status of EPIs born at a gestational age of ≥24 weeks. Multivariate logistic regression analysis indicated that lower gestational age and the need for mechanical ventilation within 72 hours after birth were independent risk factors for mortality or severe complications in EPIs (P<0.05). CONCLUSIONS:Active treatment can significantly extend the survival time of EPIs born at a gestational age of ≥24 weeks. Lower gestational age and the need for mechanical ventilation within 72 hours after birth are closely associated with poor survival outcomes in EPIs.

OBJECTIVES:To explore the predictive factors for hemodynamically significant patent ductus arteriosus (hsPDA) in preterm infants and to construct a nomogram prediction model for hsPDA occurrence in this population. METHODS:A retrospective analysis was conducted on the clinical data of preterm infants with gestational age <32 weeks diagnosed with patent ductus arteriosus (PDA) who were delivered at Nanjing Women and Children's Healthcare Hospital from January 2020 to December 2022. The subjects were divided into an hsPDA group (52 cases) and a non-hsPDA group (176 cases) based on the presence of hsPDA. Univariate analysis and multivariate logistic regression analysis were performed to screen predictive variables regarding the general information of the infants at birth, maternal pregnancy and delivery conditions, and relevant indicators during hospitalization. A nomogram prediction model for hsPDA occurrence was constructed using R software in preterm infants. Internal validation was performed using the Bootstrap method. Finally, the predictive model was evaluated for calibration, discrimination ability, and clinical utility. RESULTS:Multivariate regression analysis showed that the ratio of the left atrium to aorta diameter (LA/AO), mode of delivery (vaginal), and duration of mechanical ventilation were independent predictive factors for hsPDA in preterm infants (P<0.05). Based on the results of univariate analysis and multivariate logistic regression analysis, variables used to construct the nomogram prediction model for hsPDA risk included: LA/AO ratio, mode of delivery (vaginal), duration of mechanical ventilation, 5-minute Apgar score, and the presence of neonatal respiratory distress syndrome requiring surfactant therapy. The area under the receiver operating characteristic curve for this model was 0.876 (95%CI: 0.824-0.927), and the calibrated curve was close to the ideal reference line, indicating good calibration. The Hosmer-Lemeshow test demonstrated that the model fit well, and the clinical decision curve was above the extreme curves. CONCLUSIONS:The nomogram prediction model, constructed using five variables (LA/AO ratio, vaginal delivery, duration of mechanical ventilation, 5-minute Apgar score, and the presence of neonatal respiratory distress syndrome requiring surfactant therapy), has reference significance for predicting the occurrence of hsPDA in preterm infants and provides valuable guidance for the early clinical identification of hsPDA.

This study investigated the anti-necrotizing enterocolitis (NEC) effects of LFDP1, a lactoferrin-derived peptide previously identified as up-regulated in human milk from mothers of preterm infants. Using both in vivo and in vitro models, we evaluated LFDP1's potential to attenuate NEC development. In NEC rat pups, LFDP1 administration significantly reduced the severity of clinical symptoms and pathological features. At the cellular level, LFDP1 restored viability and enhanced migration and proliferation of IEC6 and FHC cells. Molecular analysis revealed that LFDP1 downregulated key NEC process indicators (TLR4, ACSL4, and LPCAT3) while upregulating protective factors (ZO-1 and GPX4). RNA sequencing of LFDP1-treated FHC cells identified 117 differentially expressed mRNAs, with pathway analysis highlighting the involvement of polyunsaturated fatty acid biosynthesis and fatty acid metabolism. Further investigation showed that LFDP1 suppressed ferroptosis by inhibiting the ACSL4-LPCAT3 signaling axis, a finding corroborated in human NEC intestinal samples. Notably, LFDP1 treatment reduced oxidative stress and modulated the expression of ferroptosis-related proteins in vitro. These findings suggest that LFDP1 alleviates experimental NEC by repressing polyunsaturated fatty acid production and inhibiting ferroptosis via the ACSL4-LPCAT3 pathway. Our study not only elucidates a potential mechanism for LFDP1's protective effects but also contributes to the broader understanding of how breast milk composition may protect newborns from diseases like NEC.

BACKGROUND:Previous studies have suggested that probiotics may have potential benefits for preterm infants. Their efficacy seems to depend on the particular species or combinations used. METHODS:To further investigate the effects of probiotics in preventing necrotizing enterocolitis (NEC) and other related outcomes in preterm infants, we conducted a network meta-analysis of 51 randomized controlled trials involving 11,661 participants. RESULTS:Our study revealed that most probiotics can effectively reduce the incidence of NEC (at or beyond Bell's stage II). Lactobacillus (RR, 0.59; 95% CI: 0.29, 0.98), the combination of Bifidobacterium and Lactobacillus (RR, 0.47; 95% CI: 0.20, 0.87), and the combination of Bifidobacterium, Lactobacillus, and Streptococcus (RR, 0.17; 95% CI: 0.00, 0.84) were the only treatments that significantly reduced all-cause mortality compared to placebo. Lactobacillus can be effective in reducing the time preterm infants spend in the hospital (MD, -4.23; 95% CI: -7.62, -0.81) and reaching full enteral feeding (MD, -2.15; 95% CI: -3.70, -0.64). CONCLUSIONS:The combination of Bifidobacterium, Lactobacillus, and Enterococcus was the most efficacious in reducing the mortality and incidence of NEC (Bell II or above) in preterm infants. Both prebiotics and Lactobacillus alone were found to be highly effective in reducing the length of hospitalization and the time needed to achieve full enteral feeding. No evidence suggests that probiotics affect sepsis risk. TRIAL REGISTRATION:The study protocol was registered with PROSPERO (CRD42023460231) on March 10, 2023.

Background Human milk fortifier (HMF) composition has been optimized recently. But clinical evidence of its safety and efficacy is limited in Chinese population. The aim of this study was to evaluate effects of a new HMF in growth, nutritional status, feeding intolerance, and major morbidities among very preterm (VPT) or very low birth weight (VLBW) infants in China. Methods VPT/VLBW infants admitted from March 2020 to April 2021 were prospectively included in the experimental (new HMF, nHMF) group, who received a new powdered HMF as a breast milk feeding supplement during hospitalization. Infants in the control group (cHMF) admitted from January 2018 to December 2019, were retrospective included, and matched with nHMF group infants for gestational age and birth weight. They received other kinds of commercially available HMFs. Weight gain velocity, concentrations of nutritional biomarkers, incidence of major morbidities, and measures of feeding intolerance were compared between the two groups. Results Demographic and clinical characteristics of infants in nHMF and cHMF groups were comparable. Weight gain velocity had no significant difference between the nHMF (14.0 ± 3.5 g/kg/d) and the cHMF group (14.2 ± 3.8 g/kg/d; P = 0.46). Incidence of morbidities, including necrotizing enterocolitis, bronchopulmonary dysplasia, retinopathy of prematurity, culture-confirmed sepsis, and feeding intolerance during hospitalization between nHMF and cHMF, were similar (all P -values > 0.05). The time to achieve full enteral feeding [13.5 (10, 21) days] in the nHMF group was significantly shorter than that in the cHMF group [17 (12, 23) days, HR = 0.67, 95%CI: 0.49, 0.92; P = 0.01]. Compared with cHMF group, the decrease of blood urea nitrogen level over time in nHMF group was smaller (β = 0.6, 95%CI:0.1, 1.0; P = 0.01). Conclusions The new HMF can promote growth of preterm infants effectively without increasing the incidence of major morbidity and feeding intolerance. It can be used feasible in Chinese VPT/VLBW infants. Trial registration This study was registered on ClinicalTrials.gov (NCT04283799).

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Human breast milk, the best source of nutrition for newborns, can reduce the incidence of bronchopulmonary dysplasia (BPD). Bioactive peptides are important components of human breast milk. However, their function in BPD are unclear. We screened a novel peptide (named MDABP) in the breast milk of mothers of premature infants, which has beneficial effects in experimental BPD. This study aimed to investigate the role and mechanism of MDABP in vivo and vitro. Newborn Sprague-Dawley (SD) rats were exposed to normoxia (21 % O2) or hyperoxia (85 % O2) and injected with MDABP or PBS from postnatal days 1 to 7 (PN1-PN7). On PN7, the lungs were harvested for histological and biochemical analysis. The results revealed that MDABP improved alveolar simplification and pulmonary vascular retardation, promoted alveolar epithelial cell (AEC) proliferation and inhibited cell apoptosis. In vitro, MDABP, but not scrambled MDABP, promoted cell proliferation and reduced cell injury without obvious toxicities. RNA-sequencing in A549 cells showed a total of 214 genes were significantly differentially expressed between the MDABP + hyperoxia and hyperoxia groups, including 80 upregulated and 134 downregulated genes. KEGG pathway analysis showed that significant differentially expressed genes were related to the ferroptosis signaling pathway. Rescue experiments showed that MDABP reduced the hyperoxia- and ferroptosis-induced damage to AEC by decreasing the levels of Fe2+ and ROS and increasing the levels of GSH and GPX4. The p-value of the above experiment was less than 0.05, which was considered statistically significant. This study provides a new basis for developing treatments for BPD.

Neonatal necrotizing enterocolitis (NEC) is a severe inflammatory bowel disease that commonly affects premature infants. Breastfeeding has been proven to be one of the most effective methods for preventing NEC. However, the specific role of lipids, the second major nutrient category in human breast milk (HBM), in intestinal development remains unclear. Our preliminary lipidomic analysis of the HBM lipidome revealed that dioleoyl phosphatidylethanolamine (DOPE) is not only abundant but also shows high solubility in lipids, endowing it with significant biological utility. Experimental results confirmed that DOPE significantly reduces the mortality of neonatal rats, ameliorates impairment of intestinal barrier function, and alleviates the expression of intestinal inflammatory factors IL-1β and IL-6. Furthermore, DOPE promotes the migration and proliferation of intestinal epithelial cells, thereby enhancing the integrity of the intestinal barrier function in vitro. The progression of NEC is linked with the onset of ferroptosis. Our cellular-level analysis of lipid peroxide and iron ion concentrations revealed that DOPE significantly reduces the indicators of ferroptosis, while also modulating the expression of pivotal ferroptosis-associated factors, including SLC7A11, GPX4, and ACSL4. Hence, this research on DOPE is expected to provide novel insights into the bioactive lipids present in HBM.

Necrotizing enterocolitis (NEC) is a severe inflammatory gastrointestinal disorder affecting preterm infants. Through fecal metabolomics, we found linoleic acid compounds (LAs) reduced in NEC and negatively correlated with C-reactive protein and NEC Bell stage. Conjugated linoleic acid (CLA) is a special kind of LAs with multiple functions. This study aims to investigate the role and mechanism of CLA in vivo and vitro. Experimental results in neonatal rats demonstrated that CLA significantly reduced mortality, weight loss, improved pathological scores, relieved inflammation and preserved intestinal cell homeostasis. In vitro, CLA promoted cell proliferation and reduced cell injury. RNA-sequencing analysis indicated that PPAR signaling pathway might be critical. Experiments showed that CLA facilitated PPARγ/NFκB signaling pathway by upregulating PPARγ expression, inactivating phosphorylated NFκB, and preventing translocation of NFκB P65. This study underscores the therapeutic potential of CLA and provides new insight and direction for developing NEC interventions.

Breast milk has significant benefits to preterm infants, including reduced risk of infection and inflammatory disorders. Many biomolecules, such as peptides, that exhibit immunomodulation function or antimicrobial activity and protect against infection have been identified in human milk. β-Casein 65, an endogenous peptide cleaved amino acids 204-225 of β-casein, is expressed at high levels in preterm breast milk. Our study demonstrates the antibacterial activity of β-casein 65 against Staphylococcus aureus ( S. aureus ) and Escherichia coli ( E. coli ) through use of an agarose filter paper diffusion and microdilution method. The LIVE/DEAD BacLight bacterial viability assay results also confirmed the antibacterial effects of β-casein 65. Destroyed membrane structures in S. aureus and E. coli after treatment with β-casein 65 were observed by scanning electron microscopy and transmission electron microscopy. A DNA-binding assay showed that β-casein 65 exerts antimicrobial effects independent of DNA-binding activity. Our results revealed the antibacterial activity and mechanism of peptide β-casein 65 sourced from human milk. Further studies should be carried out to determine ways to prevent and treat pathogens in animals and human neonates.