Chronic stress-induced sleep disorders are characterized by disrupted sleep architecture and a significant reduction in slow-wave sleep (SWS). These disorders represent a major public health challenge and are mechanistically linked to the hyperactivation of the hypothalamic-pituitary-adrenal (HPA) axis. This study employed a translational research paradigm-including zebrafish screening, a chronic unpredictable mild stress (CUMS) mouse model, and an eight-week randomized, placebo-controlled human trial-to evaluate the efficacy of a combination of walnut peptide and theanine (WPT). Treatment with WPT reduced waking activity and duration in pentylenetetrazole (PTZ)-induced zebrafish. In CUMS mice, the combination significantly improved sleep architecture by restoring the duration of SWS and enhanced sleep quality by increasing delta wave power density. Mechanistically, the intervention corrected hypothalamic-pituitary-adrenal (HPA) axis hyperactivity by lowering elevated serum corticosterone (CORT) levels. Furthermore, it modulated central neurotransmitters, notably reversing stress-induced deficits by increasing the levels of the inhibitory neurotransmitter GABA and tryptophan, a key precursor for serotonin and melatonin. In the human trial, the WPT restored sleep duration and improved subjective sleep quality scores (assessed by the Pittsburgh Sleep Quality Index, PSQI). In conclusion, this translational study provides robust evidence that the WPT effectively improves chronic stress-induced sleep disorders in zebrafish and mice, and improves sleep disturbances in adults (PSQI ≥ 7) by regulating HPA axis function and restoring the duration and quality of SWS. This makes it a highly promising nutritional intervention strategy.

Marginal zinc deficiency (MZD) is a common but often overlooked nutritional condition associated with impaired intestinal barrier integrity and immune dysfunction. Milk lactopeptide (LP) have attracted interest as food-derived bioactive ingredients with potential zinc-binding capacity and immunomodulatory activity. In this study, the physicochemical characteristics and predicted Zn2+-binding potential of LP were evaluated, and their effects on gut-immune disturbances in MZD rats were investigated. LP were characterized by molecular weight distribution, amino acid composition, spectroscopy, and in silico peptide screening. An MZD rat model was then established to assess the effects of graded LP intervention on growth performance, immune-related parameters, intestinal morphology, zinc status, barrier-related proteins and genes, and gut microbiota composition. The results showed that LP were rich in low-molecular-weight fractions and contained peptide features consistent with potential Zn2+ interaction. In vivo, LP improved growth retardation, partially restored immune organ development, increased blood zinc levels and apparent zinc absorption, and ameliorated colonic structural damage in a dose-dependent manner. These effects were associated with the modulation of tight junction-associated proteins and genes, as well as changes in ZIP6 expression and NF-κB-related signaling. and alterations in gut microbiota composition. Collectively, these findings suggest that fermented LP may help alleviate MZD-associated gut-immune disturbances and may serve as a promising food-derived ingredient for nutritional intervention.

Acid-precipitated casein-rich fractions from buffalo and cow milk were compared for their ability to generate dipeptidyl peptidase-IV (DPP-IV) inhibitory peptides under identical hydrolysis and fractionation conditions. Among the four proteases, Flavourzyme treatment for 3 h produced the strongest inhibition in both substrates, although it did not yield the highest degree of hydrolysis or BSA-equivalent peptide content. Spectroscopic analyses indicated substantial hydrolysis-induced changes in aromatic residue environments, surface hydrophobicity, and secondary structure. After ultrafiltration, the <3 kDa fractions showed the highest DPP-IV inhibition at the same tested concentration of freeze-dried fraction, and the buffalo-derived fraction showed higher inhibition than the corresponding cow-derived fraction. LC-MS/MS and in silico screening identified eight candidate peptides from the buffalo fraction. Four peptides were synthesized and validated, with QPPQPLPPT and LPNTVP showing the lowest IC50 values of 0.360 mg/mL (369 μM) and 0.440 mg/mL (689 μM), respectively. These findings identify acid-precipitated buffalo milk casein-rich fractions as promising precursors of DPP-IV inhibitory peptides.

This study aimed to investigate the correlation between the Triglycerides × Total Cholesterol × Body Weight Index (TCBI) and bone mineral density (BMD) in children and whether bone-specific alkaline phosphatase (B-ALP), and insulin-like growth factor 1 (IGF-1) mediate the relationship between TCBI and BMD. This study included 411 preschool children from 2 township kindergartens in a rural area of Gansu Province, China, who were surveyed at baseline in September 2021 and followed up in September 2022. The independent variable is the mean value of TCBI at baseline and follow-up. The dependent variable was the raw value of BMD during the follow-up period, measured by dual-energy X-ray absorptiometry. Multiple linear regression was used to analyze the relationship between TCBI and BMD in the nondominant forearm and heel bone. The results showed that TCBI in preschoolers was significantly and positively correlated with BMD (P < 0.001). In the fully adjusted model, for each unit increase in log-transformed TCBI, the BMD of the nondominant forearm and heel bone increased by 0.042 g/cm2 (95

The large-scale and efficient production of high quality milk-derived extracellular vesicles (mEVs) is essential to study their function and properties. The low concentrations of mEVs in milk, coupled with interference from casein, makes the enrichment of EV from large quantities of milk a major challenge. Therefore, the effects of casein removal via enzyme precipitation (EP) and acid precipitation (AP) on mEVs separation were compared in this study. Then, three concentration methods, vacuum heating concentration (VHC), vacuum freeze concentration (VFC) and membrane concentration (MC), were employed to concentrate mEVs-rich whey to improve the efficiency before ultracentrifugation. Subsequently, the morphology, particle size and marker proteins of mEVs were characterized by nanoparticle tracking analysis, transmission electron microscopy and western blotting. Finally, the function of mEVs obtained was verified by RAW264.7 cells. The results indicated that mEVs with intact morphology and higher purity could be obtained quickly through MC concentration combined with ultracentrifugation after casein removal from milk via EP, thereby providing a promising pathway for the large-scale production of milk-derived mEVs.

Stress is a prevalent mental health concern emerging predominantly in late adolescence or early adulthood. Since 2007, the Food and Drug Administration (FDA) has not approved any novel anxiolytic pharmaceuticals, fueling interest in nutritional supplements as alternative therapies for stress management. Building on prior zebrafish research, this study investigates the synergistic effects of Theanine (Th) and Walnut Peptide (WP) on stress mitigation and cognitive enhancement. Utilizing the human brain organoid stress (BO-stress) model, WP+ Th were observed to reduce stress and regulate expression of neurotransmitters, including Gamma-Aminobutyric Acid (GABA), serotonin (5-HT), dopamine (DA), and acetylcholine (Ach), as well as brain-derived neurotrophic factor (BDNF) and serotonin transporter (SERT). Subsequent in vivo study using C57BL/6J mouse-stress model demonstrated that the treatment (Th 85 mg/mL + WP 200 mg/mL), or administrated with vehicles, significantly improved their performance in stress and cognitive assessments, partially normalized neurotransmitter imbalances by modulating SERT and BDNF expression. These findings highlighted the potential of using WP + Th, particularly when delivered with vehicles (eg: powder/yogurt/milk), as a novel combined therapeutic approach for stress management and cognitive enhancement. We performed a correlation analysis between BO-stress model and mouse-stress model, revealing a alignment in the SERT levels. In addition, SERT was highly correlated with other markers in the mouse hippocampus and may represent a key target for modulating the balance between stress and cognition. ### Competing Interest Statement The authors have declared no competing interest.

BACKGROUND:Stress is a prevalent mental health concern that often emerges in late adolescence or early adulthood. Since 2007, the Food and Drug Administration (FDA) has not approved any novel anxiolytic pharmaceuticals, leading to increased interest in nutritional supplements as alternative therapies for stress management. PURPOSE:Building on our previous study, this work aims to investigate the synergistic effects of Theanine (Th) and Walnut Peptide (WP) on stress mitigation and cognitive enhancement. STUDY DESIGN:This study employed both in vitro and in vivo approaches. The former employed the human brain organoid stress (BO-stress) model, while the latter utilized the C57BL/6J mouse-stress model to evaluate the effect of Th and WP. METHODS:In the BO-stress model, the Th and WP combination was assessed for its effects on stress levels and the expression of neurotransmitters (GABA: Gamma-Aminobutyric Acid; 5-HT: serotonin; DA: dopamine; Ach: acetylcholine), as well as brain-derived neurotrophic factor (BDNF) and serotonin transporter (SERT). In the in vivo study, mice were administered Th (85 mg/ml) + WP (200 mg/ml) or with vehicles (e.g., powder, yogurt, milk), and assessed for stress and cognitive performance. Neurotransmitter levels, SERT and BDNF expression levels were also measured. RESULTS:The BO-stress model demonstrated that Th + WP reduced stress and regulated the expression of neurotransmitters and neurotrophic factors. In the mouse-stress model, the treatment improved stress and cognitive performance, partially normalizing neurotransmitter imbalances by modulating SERT and BDNF expression. We also revealed a correlation between SERT levels in the BO-stress model and those in the hippocampus of the mouse-stress model (r = 0.640). Western blot analysis further demonstrated Th + WP upregulated the expression of BDNF, TrkB (Tyrosine kinase B) and phosphorylation of CREB (cAMP response element-binding protein). CONCLUSION:In this study, we demonstrated that combined treatment with Th and WP modulates the BDNF-TrkB-CREB signaling pathway, resulting in downregulation of SERT expression and elevation of BDNF levels in the BO stress model. These molecular changes were accompanied by improved behavioral performance in mice subjected to stress and cognitive assessments. Our findings highlight the potential of Th + WP as non-pharmacological interventions for stress reduction and cognitive enhancement. Additionally, correlation analysis revealed a strong association in SERT expression between the BO and mouse stress models, further supporting cross-model consistency. SIGNIFICANCE STATEMENT:We established the novel human brain organoid-stress model, and coupled with mouse-stress model, we elucidated the synergistic actions of Th and WP in mitigating stress and augmenting cognitive function. Their effects were demonstrated to through the regulation of SERT, BDNF as well as the BDNF-TrkB-CREB pathway. This study presented a promising non-pharmacological avenue for mental health care. In addition, species differences between humans and mice were also examined through correlation analysis involving brain organoid and mouse models.

The impact of milk on bone health in rural preschoolers is under-researched. This study, through a clinical trial and a meta-analysis, finds that milk supplementation enhances forearm and calcaneus bone acquisition in children, supporting the benefits of daily milk consumption. This study evaluated the impact of dairy supplementation on bone acquisition in children’s limbs through a cluster-randomized controlled trial and a meta-analysis. The trial involved 315 children (4–6 year) from Northwest China, randomized to receive either 390 ml of milk daily (n = 215) or 20–30 g of bread (n = 100) over 12 months. We primarily assessed bone mineral density (BMD) and content (BMC) changes at the limbs, alongside bone-related biomarkers, measured at baseline, the 6th and 12th months. The meta-analysis aggregated BMD or BMC changes in the forearm/legs/calcaneus from published randomized trials involving children aged 3–18 years supplemented with dairy foods (vs. control group). Of 278 completed the trial, intention-to-treat analysis revealed significant increases in BMD (4.05

Anxiety disorders are the most common mental disorders and, without proper treatment, may lead to severe conditions: e.g., somatic disorders or permanent damage to central nervous system. Although there are drugs in clinical trials, this study focuses on exploring the efficacy of nutrients in treating these diseases. We built different zebrafish models and screened several nutrient combinations for their antianxiety, antioxidant, neuro-protecting, and memory-improving activities. Our results showed that the combinations of nutrients (e.g., Walnut Peptides + Theanine at 14.2 + 33.3 μg/ml) have similar or better activities than the positive control drugs. In addition, we discovered that the effects of the nutrients in the above four aspects were universal and highly related. This study is noteworthy as it suggested that nutrients could be healthier and greener drug alternatives and provide similar or better universal treatments for anxiety and related conditions.

Herein, we explored the effects of Poria cocos extract, protein powder mixture, and their combined intervention on weight loss in high-fat diet (HFD)-induced obese mice. Male C57BL/6J mice were selected and fed a HFD for 8 weeks; obese mice that were successfully modeled were divided into modeling and five intervention groups, and given the corresponding treatment for 10 weeks. Body weight, fat, and muscle tissue, blood glucose, lipids, inflammatory factors, and other glucose and lipid metabolism-related indicators were measured to evaluate the effect of P. cocos and protein powder intervention on weight loss in obese mice. The body weight of the intervention group was reduced compared with the HFD group. Fat content of mice in F3PM group decreased significantly (p < .05). Levels of blood glucose, lipids, adiponectin, leptin, and inflammatory factors, including interleukin-1 beta and tumor necrosis factor- alpha showed improvement. Lipoprotein lipase (lower about 2.97 pg/ml, vs. HFD mice 10.65 mmoL/ml) and sterol regulatory element-binding transcription factor (lower about 1413.63 pg/ml, vs. HFD mice 3915.33 pg/ml) levels in liver tissue were decreased. The respiratory exchange rate (RER) of mice in the HFD and subject intervention groups had no circadian rhythm and was maintained at approximately 0.80. The protein powder mixture (PM) group had the lowest RER (p < .05), the P. cocos extract (FL) and F1PM groups had similar RER to the HFD group (p < .05), and the F2PM group had a higher RER than the HFD group (p < .05). And food intake and energy metabolism returned to circadian rhythm, with an increase in the dose of P. cocos extract, the feeding rhythms of F1PM, F2PM, and F3PM were closer to that of the normal diet (ND) group. Feeding intervention with P. cocos and protein powder improved fat distribution, glucolipid metabolism, and energy metabolism, with the combination of F3PM showing more diverse benefits.

IntroductionAnxiety disorders continue to prevail as the most prevalent cluster of mental disorders following the COVID-19 pandemic, exhibiting substantial detrimental effects on individuals’ overall well-being and functioning. Even after a search spanning over a decade for novel anxiolytic compounds, none have been approved, resulting in the current anxiolytic medications being effective only for a specific subset of patients. Consequently, researchers are investigating everyday nutrients as potential alternatives to conventional medicines. Our prior study analyzed the antianxiety and memory-enhancing properties of the combination of Walnut Peptide (WP) and Casein Peptide (CP) in zebrafish.Methods and ResultsBased on this work, our current research further validates their effects in mice models exhibiting elevated anxiety levels through a combination of gavage oral administration. Our results demonstrated that at 170 + 300 mg human dose, the WP + CP combination significantly improved performances in relevant behavioral assessments related to anxiety and memory. Furthermore, our analysis revealed that the combination restores neurotransmitter dysfunction observed while monitoring Serotonin, gamma-aminobutyric acid (GABA), dopamine (DA), and acetylcholine (ACh) levels. This supplementation also elevated the expression of brain-derived neurotrophic factor mRNA, indicating protective effects against the neurological stresses of anxiety. Additionally, there were strong correlations among behavioral indicators, BDNF (brain-derived neurotrophic factor), and numerous neurotransmitters.ConclusionHence, our findings propose that the WP + CP combination holds promise as a treatment for anxiety disorder. Besides, supplementary applications are feasible when produced as powdered dietary supplements or added to common foods like powder, yogurt, or milk.

Dairy foods are crucial for adequate calcium intake in young children, but scarce data are available on the effects of formula milk on bone acquisition. This cluster-randomized controlled trial investigated the effects of the supplementation of formula milk on bone health in rural children accustomed to a low-calcium diet between September 2021 and September 2022. We recruited 196 healthy children aged 4–6 years from two kindergartens in Huining County, Northwest China. A class-based randomization was used to assign them to receive 60 g of formula milk powder containing 720 mg calcium and 4.5 µg vitamin D or 20–30 g of bread per day for 12 months, respectively. Bone mineral density (BMD) and bone mineral content (BMC) at the left forearm and calcaneus, bone biomarkers, bone-related hormones/growth factors, and body measures were determined at baseline, 6, and 12 months. A total of 174 children completed the trial and were included in the analysis. Compared with the control group, formula milk intervention showed significant extra increments in BMD (3.77% and 6.66%) and BMC (4.55% and 5.76%) at the left forearm at 6th and 12th months post-intervention (all p < 0.001), respectively. Similar trends were observed in BMD (2.83%) and BMC (2.38%) in the left calcaneus at 6 months (p < 0.05). The milk intervention (vs. control) also showed significant changes in the serum concentrations of osteocalcin level (−7.59%, p = 0.012), 25-hydroxy-vitamin-D (+5.54%, p = 0.001), parathyroid hormone concentration (−15.22%, p = 0.003), and insulin-like growth factor 1 (+8.36%, p = 0.014). The percentage increases in height were 0.34%, 0.45%, and 0.42% higher in the milk group than in the control group after 3-, 6-, and 9-month intervention, respectively (p < 0.05). In summary, formula milk supplementation enhances bone acquisition at the left forearm in young Chinese children.

ObjectivesObesity is often associated with glucolipid and/or energy metabolism disorders. Ascophyllum nodosum extract (seaweed extract, SE) and Camellia sinensis-leaf extract (tea extract, TE) have been reported to promote positive metabolic effects through different mechanisms. We investigated the effects of SE and TE on metabolic homeostasis in diet-induced obese mice and discussed their functional characteristics.MethodsMale C57BL/6J mice fed with high-fat diets for 8 weeks were established as obese models and subsequently divided into different intervention groups, followed by SE, TE, and their joint interventions for 10 weeks. Body weight and food intake were monitored. Fasting glucose and oral glucose tolerance tests were interspersed during the experiment. After the intervention, the effects on obesity control were assessed based on body composition, liver pathology section, blood lipids and glucose, respiratory exchange ratio (RER), energy expenditure (EE1, EE2, and EE3), inflammatory factors, lipid anabolism enzymes, and gut flora of the obese mice.ResultsAfter continuous gavage intervention, the mice in the intervention groups exhibited lower body weight (lower ~4.93 g, vs. HFD 38.02 g), peri-testicular fat masses (lower ~0.61 g, vs. HFD 1.92 g), and perirenal fat masses (lower ~0.21 g, vs. HFD mice 0.70 g). All interventions prevented diet-induced increases in plasma levels of glucose, adiponectin, leptin, and the inflammatory factors IL-1β and TNF-α. The RER was modified by the interventions, while the rhythm of the RER was not. Blood lipids (total cholesterol, triglycerides, and LDL) decreased and were associated with lower lipid anabolism enzymes. In addition, the SE and TE interventions altered the structure and abundance of specific flora. Different interventions inhibited the growth of different genera positively associated with obesity (Escherichia–Shigella, Helicobacter, etc.) and promoted the growth of Akkermansia and Bacteroides, thus affecting the chronic inflammatory state.ConclusionSE and TE both have synergistic effects on weight control and glucolipid metabolism regulation by improving insulin sensitivity and reducing lipid synthesis-related enzyme expression, whereas the combination of SE and TE (3:1) has a better effect on regulating energy metabolism and inhibiting chronic inflammation.

Coix seed extract (CSE) and probiotics have been reported to regulate glycolipid metabolism through different modes of action. We tested the effects of CSE, Lactobacillus paracasei K56, and their combination to determine whether they have synergistic effects on glycolipid metabolism of obese mice. We fed male C57BL/6J mice with high-fat diet for 8 weeks to establish an obesity model. The obesity mice were selected and divided into five groups: the model control group and four intervention groups. After 10 weeks of continuous gavage intervention, the mice in the intervention groups exhibited lower body weight (lower about 2.31-4.41 g, vs. HFD 42.25 g, p < 0.01), and epididymal (lower about 0.58-0.92 g, vs. HFD 2.50 g, p < 0.01) and perirenal fat content (lower about 0.24-0.42 g, vs. HFD 0.88 g, p < 0.05); decreased fasting blood glucose, total cholesterol, triglycerides, and VLDL; and increased HLDL, respiratory exchange ratio, energy expenditure, and amount of exercise performed. K56 + CSE-combined intervention groups were more effective in lowering blood glucose, IL-1β, and TNF-α levels than the CSE and K56 alone interventions. The content of fatty acid synthase and SREBP-1c protein in liver tissue was lower. The combination has synergistic effects on weight control, fat reduction, and blood glucose regulation by improving the chronic inflammatory state and reducing the content of lipid synthesis-related enzymes of obese mice, which can hinder chronic disease progression. PRACTICAL APPLICATION: Coix seed extract can be used in obese people to regulate abnormal glucose and lipid metabolism and delay the development of chronic diseases.

IntroductionBrain development during infancy is crucial for later health and development. Although Milk Fat Globule Membrane (MFGM) has been demonstrated to enhance brain development, further investigation is needed to determine the optimal dose.MethodsIn this study, 80 piglets aged 2 days were randomly assigned to four groups: Control group, MFGM-L (1.74 g MFGM per 100 g diet), MFGM-M (4.64 g MFGM per 100 g diet), and MFGM-H (6.09 g MFGM per 100 g diet). Daily body weight and milk intake of the piglets were recorded until 31 days postnatal. Learning and memory abilities were evaluated using the spatial T-maze test on day 15. MRI analysis was conducted to assess functional and structural changes in brain tissues. Additionally, mRNA and protein expression of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NTF-3) in the hippocampus and prefrontal cortex were evaluated.ResultsThe results indicated that the MFGM supplemented diet significantly improved the accuracy of the piglets in the T-maze test, with the MFGM-L group exhibiting the best performance. MRI showed no volumetric differences in the gray and white matter between the groups. However, the fractional anisotropy in the left and right hippocampus of piglets in the MFGM-L group was significantly higher than in the other three groups. Furthermore, there was a strong correlation between the accuracy of the T-maze test and hippocampal fractional anisotropy.DiscussionThe MFGM supplemented diet also increased the expression of BDNF in the cerebral cortex. However, the changes in BDNF were not consistent with the results of the T-maze test. In conclusion, adding 1.74 g MFGM per 100 g diet can significantly improve neonatal piglets’ learning and memory abilities, potentially by enhancing the connection of white matter fiber bundles in the brain.

目的:测定健康人群摄入3种口服营养补充剂(ONS)的血糖生成指数(GⅠ)和胰岛素指数(Ⅱ).方法:对12名健康成人进行了单盲交叉研究.受试者在每次访视时空腹摄入服用3种受试产品中的1种,2次访视之间有1周的洗脱期.每个受试者都摄入了所有受试产品.3种受试产品分别为两种新开发的ONS(全营养粉1、全营养粉2)以及阳性对照(全营养粉3).受试者每次摄入相当于含碳水化合物50 g的受试产品,测量餐后15、30、45、60、90、120、180、240 min的血糖和胰岛素水平.用Wolever法计算血糖和胰岛素曲线下面积(AUC),得到3种营养粉剂的GⅠ和Ⅱ.结果:以葡萄糖的GⅠ、Ⅱ为参考标准,3种营养粉的GⅠ分别为44、54、29;Ⅱ分别为163、182、119.结论:两种研发的全营养粉中,全营养粉1更适合作为糖尿病患者的口服营养补充选择.

Coix seed extract (CSE) and probiotics have been reported to regulate glycolipid metabolism via different modes of action. We tested the effects of CSE, Bifidobacterium BPL1, and their combination to determine their effects on glycolipid metabolism in obese mice. Male C57BL/6J mice were fed a high-fat diet for 8 weeks to establish an obesity model. Obese mice were selected and divided into four groups: the model control group and three intervention groups. After 10 weeks of continuous gavage intervention, the mice in the intervention groups exhibited lower body weight (lower about 2.31 g, vs. HFD mice 42.23 g) and epididymal (lower about 0.37 g, vs. HFD mice 2.5 g) and perirenal fat content (lower about 0.47 g, vs. HFD mice 0.884 g); decreased fasting blood glucose, total cholesterol, triglycerides, and VLDL; and increased HLDL, respiratory exchange ratio, energy expenditure, and amount of exercise performed. CSE, BPL1 and their combination can effectively control the weight gain in obese mice, reduce fat content, and regulate blood lipids and abnormal blood sugar. These results may be related to reduce the chronic inflammatory states, improve energy metabolism, exercise, relieve insulin sensitivity, and reduce lipid synthesis via the intervention of CSE, BPL1 and their combination. Compared with the single use of CSE alone, the combination of CSE + BPL1 can better exert the regulation function of intestinal flora, and change in the abundance of bacteria that could improve the level of inflammatory factors, such as increasing Bifidobacterium, reducing Lactococcus. Compared with the use of BPL1 alone, the combination of CSE and BPL1 can better regulate pancreatic islet and improve blood sugar. CSE may act directly on body tissues to exert anti-inflammatory effects. BPL1 and CSE + BPL1 may improve the structure and function of the intestinal flora, and reduce tissue inflammation.

目的 探究低聚半乳糖(galacto-oligosaccharides,GOS)与聚葡萄糖(polydextroses,PD)联用对生长期大鼠骨代谢和骨钙含量的影响.方法 60只刚断乳SD大鼠随机分为低钙组(饲料钙0.15％)、正常组(饲料钙0.5％)、强化组(饲料钙0.375％+维生素D强化)、低比组(强化组饲料+PD 2.5％+GOS 2.5％)和高比组(强化组饲料+PD 4％+GOS 1％)5个组,分别饲喂含钙、维生素D3和益生元含量不同的饲料.观察大鼠一般情况,记录体重,于实验第83 d进行3 d的钙代谢实验,原子吸收法检测饲料、粪便和尿液中的钙含量,计算钙表观吸收率和储备率;喂饲93 d后处死,取心肝脾肾称重计算脏器系数,取血清测定血清钙、磷水平,取股骨测股骨干重,并采用原子吸收法测量股骨钙含量.结果 实验结束时,正常组、低比组和高比组体重均显著高于低钙组;各组的钙表观吸收率和钙储留率显著低于低钙组;与低钙组比较,各组的血清钙和骨钙含量都显著增加;低比组和高比组的股骨重和骨钙含量显著高于强化组,与正常组无显著差异.结论 充足的钙摄入对生长期大鼠的钙代谢及骨骼生长具有重要意义;聚葡萄糖和低聚半乳糖联用可增加生长期大鼠的骨钙含量.

BACKGROUND γ-aminobutyric acid (GABA) is a naturally occurring non-protein amino acid in the nervous system and has a wide range of physiological functions in the body. Walnut peptide (WP) contains high levels of arginine, aspartic acid, and glutamate, and has been shown to improve cognitive deficits and memory impairment in mice, while restoring antioxidant enzyme levels and reducing brain inflammatory mediators. METHODS This study investigated the effects of GABA and WP, either alone or in combination, on sleep disturbances in mice. The pentobarbital-prolonged sleep test, pentobarbital-threshold sleep test, and barbital-induced sleep test were conducted to assess the effects of GABA and WP on sleep quality by gavage for 30 days as follows: GABA (102.25 mg/kg), WP (102.25 mg/kg), GABA (33.95, 102.25, 306.75 mg/kg)/WP (102.25 mg/kg) mixture. Furthermore, neurotransmitter tests were performed using mice brain tissue to investigate the possible mechanisms of GABA and WP on sleep status. RESULTS The results showed that the combined use of GABA and WP significantly increased sleep duration compared with single administration of either WP or GABA. Increasing doses of GABA in mice treated with combined GABA and WP elevated the sleep rate to 50.00%, 64.28%, and 64.28%, respectively, compared to mice treated with GABA alone (35.71%) or mice treated with WP alone (28.57%). In mice that received a combination of GABA and WP orally, the latency time was significantly decreased after 30 days compared to control mice (P<0.05). Additionally, in mice treated with GABA, WP, or the combination of GABA and WP, the concentrations of GABA and acetylcholine (Ach) in the brain were significantly elevated and the concentration of serotonin (5-HT) was decreased compared to untreated mice. CONCLUSIONS These results demonstrated that the combined administration of GABA and WP could prolong the sleep duration, increase sleep rate, and shorten the sleep latency more effectively than the administration of either GABA or WP alone. The mechanisms of action may be related to the regulation of neurotransmitters in the brain tissue by the combination of GABA and WP.