[Effect of high selenium on insulin signaling pathway PI3K-AKT-mTOR in L02 cells].

OBJECTIVE: To observe the effect of high selenium on insulin signaling pathway PI3K-AKT-mTOR in L02 cells. METHODS: One group of L02 cell was treated with different concentrations of selenomethionine(SeMet, 0.001, 0.0025, 0.005, 0.0075, 0.01, 0.025, 0.05, 0.075 and 0.1µmol/L) for 48 h, then cultured with serum-free medium for 4 h and stimulated with 1 µmol/L insulin for 15 min. The insulin signaling pathway(PI3K-AKT-mTOR) was detected by WB. Another group of L02 cell was treated with the same concentrations of SeMet as above for 48 h. The cell supernatant and lysates were collected for the analysis of SELENOP and GPX1, respectively by WB. RESULTS: The expressions of P-AKT-(Ser-473), P-AKT-(Thr-308), PI3K and mTOR in L02 cells under high-Se were decreased with the increase of SeMet concentration. The expressions of GPX1 and SELENOP were enhanced with the increase of SeMet. CONCLUSION: The insulin signaling pathway, PI3K-AKT-mTOR, was damaged in L02 cell under high-Se stress.

[Study of high selenium interfering with glucose and one-carbon metabolism in hepatocytes in vitro].

OBJECTIVE: To investigate the effects of high selenium environment on the expression of selenoproteins and enzymes related to glucose and one-carbon metabolism in normal human hepatocytes. METHODS: Ten different concentrations of selenomethionine(SeMet, 0, 0.001, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5 and 10 µmol/L) was added into the normal human hepatocyts and incubated for 48 hours. The expressions of selenoprotein(GPX1 and SELENOP1) and metabolic enzymes(PHGDH, SHMT1, MTHFR and MS) were analyzed by Western blot. RESULTS: When the concentration of SeMet was 0-10 µmol/L, the expression trend of selenoprotein(GPX1 and SELENOP1) is similar, which first increases and then decreases. There is a slight difference between the inflection points of GPX1 and SELENOP1, which are respectively 0.5 µmol/L and 0.1 µmol/L. The expression trend of serine de novo synthesis pathway key enzymes(PHGDH) and folate cycle metabolizing enzymes(SHMT1, MTHFR and MS) is similar to that of selenoproteins, which also increases first and then decreases, but the inflection points are different, which are respectively 0.1 µmol/L(PHGDH and SHMT1) and 0.01 µmol/L(MTHFR and MS). CONCLUSION: Under the high selenium environment, the glycolytic bypass-serine de novo synthesis pathway is activated to synthesize endogenous serine due to the insufficient intracellular serine supply, causing abnormal glucose metabolism, which is an important extension to the hypothesis of the molecular mechanism of high selenium causing IR.

[Intervention study to inhibit the glucose metabolic remodeling in hepatocytes induced by high-selenium in vitro].

OBJECTIVE: To observe the effect of exogenous serine or glycine on the synthesis of selenoprotein and endogenous serine and the expression of metabolic enzymes in hepatocytes cultured with high-selenium in vitro and its dose-response relationship. METHODS: The experiment was divided into two parts, namely a inhibition experiment and a dose-response experiment, using L02 cells as the intervention target. In the inhibition experiment, the blank control group, high-Se(SeMet) group, serine intervention group and high-Se+serine intervention group were set up. Both SeMet and serine were given at a level of 0.05 µmol/L, and the blank control group was given the same volumes of saline. In the dose-response experiment, the concentration of SeMet was 0.05 µmol/L, and the intervention concentration gradients of serine or glycine were 0, 0.05, 0.1, 0.5, 1, 5, 10, 50, 100 and 500 µmol/L. The expression of phosphoglycerate dehydrogenase(PHGDH)、serine hydroxymethyltransferase 1(SHMT1)、methylenetetrahydrofolate reductase(MTHFR)、selenoprotein P(SELENOP) and glutathione peroxidase 1(GPX1)was detected by Western Blot(WB). RESULTS: (1)In the inhibition experiment, compared with the blank control group, the expression of selenium proteins(GPX1 and SELENOP) in L02 cells of the other three groups were significantly increased(P<0.05). Compared with the high expression of PHGDH in L02 cells of high-Se group, the expressions of PHGDH, SHMT1 and MTHFR in high-Se + serine group were significantly decreased(P<0.05). (2) In the dose-response experiment, the expression of PHGDH enzyme in L02 cells gradually decreased with the increase of the concentration of exogenous serine or glycine, showing an obvious dose-dependent effect. In contrast, none of the other metabolic enzymes(SHMT1 and MTHFR) showed similar trends in protein expression. CONCLUSION: The upregulated expression of PHGDH, the key enzyme in the de novo synthesis pathway of serine in hepatocytes cultured with high-selenium can be inhibited feedback by exogenous serine or endogenous serine transformed from exogenous glycine directly.

Dietary curcumin supplementation relieves hydrogen peroxide-induced testicular injury by antioxidant and anti-apoptotic effects in roosters.

This study was aimed to investigate the effects of dietary curcumin supplementation on the hydrogen peroxide (H2O2)-induced testicular oxidative damage in breeder roosters. Thirty-two 20-week roosters were randomly divided into four groups: (1) basal diet (CON); (2) basal diet with H2O2 challenge (H2O2); (3) basal diet with 200 mg/kg curcumin (CUR); (4) basal diet with 200 mg/kg curcumin and H2O2 challenge (CUR + H2O2). The trial lasted for 8 weeks, H2O2 challenged groups got an intraperitoneal injection of H2O2 at the 50 and 53 days, while the CON and CUR groups received an injection of saline. The results showed that dietary curcumin supplementation significantly decreased abnormal sperm rates in the semen, notably improved seminiferous tubules, increased testis scores, and serum testosterone levels. Curcumin supplementation could also ameliorate the redox damage caused by H2O2, by enhancing the capacities of antioxidant enzymes (CAT, GSH-Px, SOD, and T-AOC), and reducing MDA levels. In addition, curcumin normalized the H2O2-induced negative effects, which included downregulations in spermatogenesis-related genes (STAR, HSD3-ß1, SYCP3, AKT1) and antioxidant genes (HMOX-1, NQO-1), reduced protein expressions of Nrf2, PCNA, and Bcl-2, and increased protein expressions of Caspase 3 and Bax. Moreover, H2O2-induced decreased mRNA expressions of EIF2AK3, Caspase3, and BCL-2 were all reversed by dietary curcumin supplementation. In summary, dietary curcumin supplementation could relieve H2O2-induced oxidative damage and reproduction decline through the Nrf2 signaling pathway and anti-apoptotic effects in roosters.

Dietary folic acid supplementation improves semen quality and spermatogenesis through altering autophagy and histone methylation in the testis of aged broiler breeder roosters.

The aging phenomenon often exerts a significant reduction in the reproduction performance of aged animals. The objective of this project was to investigate the effects of dietary Folic acid (FA) supplementation on the reproductive performance of aged broiler breeder roosters. A total of 16 aged ROSS 308 broiler breeder roosters (50-week-old) were randomly divided into two groups. The treatments were basal diet (CON), a basal diet supplemented with 10 mg/kg Folic acid (FAS) for four weeks. At the end of the experiment, semen quality, histopathological studies, serum concentrations of testosterone and relative mRNA and protein expressions of testes were evaluated. The results showed that dietary FA supplementation dramatically improved semen quality of aged roosters, manifested by increasing semen volume, sperm concentration, sperm motility, and sperm membrane functional integrity. Furthermore, seminiferous tubule epithelial height (SEH) and testis scores were increased by dietary supplementation with FA. Dietary FA also remarkably augmented the transcription level of spermatogenesis-related gene (CREM, PCK2, DDX4, and GDNF). No significant differences were observed in serum concentrations of testosterone between FAS and CON groups. We noted significant upregulation Beclin-1 and ATG5 protein expressions, and the ratio of LC3-Ⅱ/Ⅰ, as well as significant downregulation of p-mTOR protein expressions in testicular tissue of aged roosters with FA supplementation. In addition, dietary FA supplementation significantly increased the protein expression of H3K9me2 and reduced the protein expression of H3K27me2. In summary, dietary FA supplementation improved the testicular autophagy through the mTOR-signaling pathway, and altered histone methylation in the testis. Dietary supplementation with FA can ameliorate semen quality and spermatogenesis of aged roosters.

Dietary genistein supplementation alters mRNA expression profile and alternative splicing signature in the thymus of chicks with lipopolysaccharide challenge.

Genistein is abundant in the soybean products, which exerts prominent effects on immune function. Little information is available about the effect of dietary genistein on thymic transcriptome, especially when suffering from lipopolysaccharide challenge. In this study, 180 one-day-old male broilers were randomly allocated to 3 groups: nonchallenged chicks given a basal diet (CON), and lipopolysaccharide-challenged chicks fed a basal diet (LPS), or lipopolysaccharide-challenged chicks fed a basal diet supplemented with 40 mg/kg genistein (GEN). Lipopolysaccharide injection induced thymocyte apoptosis and inflammatory reactions in the chicks. The results showed dietary genistein significantly reduced the percentage of CD3+ T lymphocytes by 10.04% and CD4+/CD8+ T lymphocyte ratio by 21.88% in the peripheral blood induced by lipopolysaccharide injection (P < 0.05). In addition, genistein significantly reduced the thymus index by 50% and apoptotic index by 12.34% induced by LPS challenge (P < 0.05). Transcriptomic analysis identified 1,926 DEGs (1,014 upregulated and 912 downregulated, P < 0.05) between GEN and LPS groups, which altered the mRNA expression profile and signaling pathways (Toll-like receptor, and NOD-like receptor signaling pathway) in the thymus. Furthermore, 5 splicing (AS) isoforms of the Drosophila Disabled-2 (DAB2) gene were detected, which were significantly upregulated in the GEN group compared with that in the LPS group. In summary, dietary genistein supplementation altered the RNA expression profile and AS signatures in the thymus, and alleviated immune response against lipopolysaccharide challenge.

Dietary mulberry-leaf flavonoids supplementation improves liver lipid metabolism and ovarian function of aged breeder hens.

Mulberry-leaf flavonoids (MF), extracted from mulberry leaves, exert antioxidant and hypolipidemic effects. The purpose of this experimental study was to investigate the effects of dietary MF on the ovarian function and liver lipid metabolism of aged breeder hens. We used 270 (60-weeks-old) Qiling breeder hens randomly assigned in 3 treatments with supplemental dietary MF doses (0, 30, 60 mg/kg). The results showed that dietary MF significantly improved the egg-laying rate, followed by the reduced feed conversion rate (FCR) (p < 0.05). However, there is no obvious difference in hatchability and fertilised eggs hatchability among the three groups (p > 0.05). The level of T-CHO, LDL-C and AKP in serum was reduced, and the HDL-C concentrations were increased by dietary MF (p < 0.05). MF treatment also improved the antioxidant capacity and reduced the apoptotic index of the ovary (p < 0.05). Additionally, dietary MF significantly increased the serum estradiol (E2) levels (p < 0.05) and the transcription level of CYP19A1 and LHR in the ovary (p < 0.05). Dietary MF enhanced fatty acid ß-oxidation in the liver via up-regulating the mRNA expressions of PPARα and CPT-I (p < 0.05). Moreover, the HMF group significantly decreased mRNA expressions of SREBP-1c (p < 0.05) and increased mRNA expressions of ERα, VTG-Ⅱ and ApoB in the liver (p < 0.05). In conclusion, dietary MF could improve the reproduction performance of aged breeder hens through improving ovary function and hepatic lipid metabolism.

The Possible Mechanism of Physiological Adaptation to the Low-Se Diet and Its Health Risk in the Traditional Endemic Areas of Keshan Diseases.

Selenium is an essential trace element for humans and animals. As with oxygen and sulfur, etc., it belongs to the sixth main group of the periodic table of elements. Therefore, the corresponding amino acids, such as selenocysteine (Sec), serine (Ser), and cysteine (Cys), have similar spatial structure, physical, and chemical properties. In this review, we focus on the neglected but key role of serine in a possible mechanism of the physiological adaptation to Se-deficiency in human beings with an adequate intake of dietary protein: the insertion of Cys in place of Sec during the translation of selenoproteins dependent on the Sec insertion sequence element in the 3'UTR of mRNA at the UGA codon through a novel serine-dependent pathway for the de novo synthesis of the Cys-tRNA[Ser]Sec, similar to Sec-tRNA[Ser]Sec. We also discuss the important roles of serine in the metabolism of selenium directly or indirectly via GSH, and the maintenance of selenium homostasis regulated through the methylation modification of Sec-tRNA[Ser]Sec at the position 34U by SAM. Finally, we propose a hypothesis to explain why Keshan disease has gradually disappeared in China and predict the potential health risk of the human body in the physiological adaptation state of low selenium based on the results of animal experiments.

Dietary mulberry-leaf flavonoids improve the eggshell quality of aged breeder hens.

Eggshell quality is subject to a significant decline in the late laying period, which results in huge economic losses. The purpose of this study was to investigate the effects of dietary mulberry-leaf flavonoids (MF) on the eggshell quality of aged breeder hens. A total of 270 (60-week-old) Qiling breeder hens were randomly assigned to 3 treatments with supplemental dietary MF doses (0, 30, and 60 mg/kg). The results showed that dietary MF improved the eggshell thickness and strength, following the reduced broken egg ratio (P < 0.05). Histological analysis showed that dietary MF increased glandular density and luminal epithelium height in the shell gland (P < 0.05). MF treatment reduced the apoptotic index of the shell gland, following by improved antioxidant capacity (P < 0.05). The protein expression of Caspase 3 was down-regulated, and Nrf2 was up-regulated by dietary MF (P < 0.05). Furthermore, calcium (Ca) content in the serum and shell gland, as well as the activity of Ca2+-ATPase in the shell gland were increased by dietary MF (P < 0.05). Ca transport-related genes (ESRα, ESRß, KCNA1, OPN, CABP-28K and CDH6) in the shell gland were upregulated by dietary MF treatment (P < 0.05). In conclusion, dietary MF could ameliorate the eggshell quality of aged hens by improving antioxidative capability and Ca deposition in the shell gland of uterus.

Dietary hawthorn-leaves flavonoids improves ovarian function and liver lipid metabolism in aged breeder hens.

Hawthorn-leaves flavonoids (HF), extracted from hawthorn leaves, were reported to exert antioxidant, anti-inflammatory and hypolipidemic properties. The aim of our study was to investigate the effects of dietary HF on the reproduction performance and liver lipid metabolism of aged breeder hens. A total of 270 aged Qiling breeder hens (60-wk-old) were randomly divided into 3 treatments: 1) basic corn-soybean diet (CON); 2) basic corn-soybean diet supplemented with 30 mg/kg HF (LHF); 3) basic corn-soybean diet supplemented with 60 mg/kg HF (HHF). The results showed that supplemented HF significantly improved the egg-laying rate and hatching rate of aged breeder hens (P < 0.05). HF treatment reduced the serum TG, T-CHO and L-LDL levels (P < 0.05), and upregulated the mRNA expressions of ESR1, ESR2, VTGⅡ, ApoB, and ApoVI in the liver (P < 0.05). Serum estrogen levels in HF treated groups were elevated compared with the CON group (P < 0.05). In the HHF group, the number of the primordial follicles was higher in comparison with the CON group (P < 0.05). Furthermore, dietary supplementation with HF improved the activity of antioxidant enzymes (T-AOC, GSH-Pχ) (P < 0.05), following with the reversed ovarian apoptosis and morphological damage. In addition, 60 mg/kg dietary HF upregulated the protein expression of PCNA and Nrf2 in the ovary (P < 0.05). In summary, dietary supplementation with HF could improve the reproduction performance through regulating liver lipid metabolism and improving ovarian function in aged breeder hens.

[Study of selenomethionine on the selenoproteins expression in hepatocyte L02 and the synergistic effect of serine].

OBJECTIVE: To observe the effect of selenomethionine(SeMet)on the selenoproteins expression in hepatocyte L02 and the synergistic effect of serine. METHODS: The L02 cells were cultured and divided into SeMet group and Serine+SeMet group. SeMet dose was set as 0. 001, 0. 01, 0. 1, 1 and 10 µmol/L. Serine and SeMet were mixed according to 2∶1 molar ratio(Serine∶SeMet=2∶1). The L02 cells were cultured for 48 h after SeMet and Serine added. Finally, the cell culture supernatants and homogenates were collected for the selenoprotein P(SEPP)and glutathione peroxidase 1(GPx1)concentrations detection by a double-antibody sandwich enzyme-linked immuno-sorbent assay(ELISA). The expression of SEPP and GPx1 in cell homogenates was detected by western blot(WB). RESULTS: ELISA and WB results showed that GPx1 and SEPP expressions were dose dependent in a low SeMet concentration range, reached their inflection points when SeMet concentration was 1 µmol/L and 0. 1 µmol/L respectively, and began to decrease when SeMet concentration was further increased to 10 µmol/L. WB result showed that serine had a synergistic effect on GPx1 and SEPP expressions as SeMet concentration was 0. 001 µmol/L to 10 µmol/L. CONCLUSION: There was an inflection point in selenium protein synthesis when SeMet acted on normal hepatocyte(L02), the specific mechanism needs to be further explored. Secondly, serine had a synergistic effect on GPx1 and SEPP expressions of SeMet in hepatocyte(L02).

Low-Se Diet Can Affect Sperm Quality and Testicular Glutathione Peroxidase-4 activity in Rats.

This study aimed to observe the influence of selenium (Se) deficiency on sperm quality and selenoprotein expression in rats. Four-week male Wista rats were randomly divided into three groups: Se-A, Se-L, and Se-D (respectively for Se- adequate, low, and deficient group). After 9 weeks, the rats were sacrificed by anesthesia, with the cauda epididymidis quickly fetched for sperm count, motility, and deformity. Meanwhile the blood, liver, brain, heart, and testis were collected for Se and biochemical analysis. It was found that the rats in Se-D had poor growth, while the Se concentrations in blood, liver, and heart for Se-D decreased significantly, compared with Se-A and Se-L (p < 0.01). But no significant difference was observed in testis and brain and also no statistical significance for sperm count. The sperm motility for Se-A (63.07%) was significantly higher than Se-L (53.91%) and Se-D (54.15%). Deformities were observed in both Se-L and Se-D. Both glutathione peroxidases (GPxs) and selenoprotein-P (SEPP1) levels in plasma and tissues of Se-D were significantly lower than those of Se-A and Se-L (p < 0.01). The SEPP1 levels in heart and brain of Se-L were lower than Se-A (p < 0.01). There was no statistical difference for GPx1 between Se-A and Se-L. The GPx4 level in testis of Se-L was lower than Se-A (p < 0.05). However, the SEPP1 in plasma, liver, testis, and the GPx3 level in plasma of Se-L were higher than those of Se-A (p < 0.05 or p < 0.01). Our results show that dietary Se deficiency could reduce GPx4 and SEPP1 expression in testis, which further influence sperm motility and may cause sperm deformity. Selenoprotein expression in some tissues of Se-L was higher than that of Se-A, but sperm quality and GPx4 expression in testis were not improved for Se-L. Low active pseudoselenoproteins might be synthesized in low-Se condition. The underlying mechanism needs to be further investigated.

Low selenium intake is associated with postpartum weight retention in Chinese women and impaired physical development of their offspring.

The aim of this study was to investigate the association between daily Se intake and postpartum weight retention (PPWR) among Chinese lactating women, and the impact of their Se nutritional status on infants' physical development. Se contents in breast milk and plasma collected from 264 lactating Chinese women at the 42nd day postpartum were analysed with inductively coupled plasma MS. Daily Se intake was calculated based on plasma Se concentration. The dietary data of 24-h records on three consecutive days were collected. Infant growth status was evaluated with WHO standards by Z-scores. Linear regression analyses and multinomial logistic regression were conducted to examine the impact of Se disequilibrium (including other factors) on PPWR and growth of infants, respectively. The results indicated that: (1) the daily Se intake of the subjects was negatively associated with their PPWR (B = -0·002, 95 % CI - 0·003, 0·000, P = 0·039); (2) both insufficient Se daily intake (B = -0·001, OR 0·999, 95 % CI 0·998, 1·000, P = 0·014) and low level of Se in milk (B = -0·025, OR 0·975, 95 % CI 0·951, 0·999, P = 0·021) had potential associations with their infants' wasting, and low level of Se in milk (B = -0·159, OR 0·853, 95 % CI 0·743, 0·980, P = 0·024) had a significant association with their infants' overweight. In conclusion, the insufficient Se nutritional status of lactating Chinese women was first found as one possible influencing factor of their PPWR as well as low physical development of their offspring.

Dietary alpha-lipoic acid supplementation improves spermatogenesis and semen quality via antioxidant and anti-apoptotic effects in aged breeder roosters.

The purpose of the present study was to investigate the effects of dietary alpha-lipoic acid (ALA) supplementation on the reproductive performance of aged breeder roosters. Sixteen 50-wk-old ROSS 308 breeder roosters were randomly allocated to two groups: roosters received a basal diet (CON), or a basal diet supplemented with 300 mg/kg of ALA (ALA). The results indicated that dietary ALA supplementation significantly increased sperm concentration, motility, viability, and membrane functional integrity. ALA also dramatically increased seminiferous tubule epithelial height (SEH) and testis scores. The ALA group had a higher serum concentration of testosterone than the CON group. ALA supplementation remarkably increased total antioxidant capacity (T-AOC), the enzyme activities of glutathione peroxidase (GPx), and catalase (CAT) in the testes; following a decrease in malondialdehyde (MDA) levels. In addition, we noted significant upregulation of Nrf2 mRNA and protein expression of and mRNA expression of its Downstream Genes (GPx1, NQO1, and GCLC), as well as significant downregulation of Keap1 mRNA expression in testicular tissue of aged roosters with ALA supplementation. The protein expression of Caspase 3 was downregulated and the protein expression of proliferating cell nuclear antigen (PCNA) was upregulated by ALA supplementation. The mRNA expression of spermatogenesis-related genes (ER1, AKT1, and Cav1) were markedly augmented in the ALA group compared with the CON group. In conclusion, dietary ALA supplementation enhanced the testicular antioxidant capacity through the Nrf2-signaling pathway, exerted anti-apoptotic effects, and improved the reproductive performance of aged roosters.

Dietary Serine and Sulfate-Containing Amino Acids Related to the Nutritional Status of Selenium in Lactating Chinese Women.

Previous studies suggested that serine can promote the synthesis of selenoproteins and the interaction, transformation, and replacement of serine, cysteine, and selenocysteine have been observed in the human body. This study was designed to clarify whether the dietary intakes of serine and sulfate-containing amino acids (SAAs) could directly affect the selenium (Se) nutritional status or the level of milk Se in lactating women. Breast milk and plasma samples were collected from a total of 264 lactating Chinese women when they revisited their local hospital at the 42nd day postpartum to detect the concentration of Se with ICP-MS and the content of selenoprotein P (SEPP1) and the activity of glutathione peroxidase 3 (GPX3) in the plasma by ELISA. The daily Se intake by each subject was calculated based on her own plasma Se concentration. The 24-h dietary record data for 3 consecutive days were collected to calculate their dietary intakes of protein together with each amino acid daily based on the China Food Composition Tables (CFCT). Ordinal polytomous logistic regression was applied to examine the determinants of BMI values for lactating women. For all subjects, the concentration of plasma SEPP1 and milk Se of participants with insufficient Se intake were significantly associated with the dietary intake of serine and 2 SAAs (methionine and cystine), respectively (P < 0.05). No significant correlation was found between each amino acid related to the synthesis of endogenous serine and every biomarker of the Se nutrition status in subjects with an insufficient dietary protein intake (P > 0.05). The ordinal logistic regression analysis showed that dietary protein intake (ordinal OR 1.012, 95% CI = 0.004-0.020, P = 0.002) and plasma SEPP1 (ordinal OR 0.988, 95% CI = - 0.023 to - 0.001, P = 0.036) affected the BMI value together in these lactating women. In conclusion, dietary serine and SAAs were found to directly affect the nutritional status, and both high protein intake and low plasma SEPP1 might be the health risks in these lactating Chinese women.

Breast milk selenocystine as a biomarker for selenium intake in lactating women at differential geographical deficiency risk in China.

BACKGROUND AND OBJECTIVES: A reliable biomarker for optimal selenium (Se) intake in lactating women is not currently available. METHODS AND STUDY DESIGN: Daily dietary Se intake in lactating women was calculated from a 24-hour meal record survey for over 3 days. Se levels in plasma and breast milk were measured through inductively coupled plasma mass spectrometry. Plasma selenoprotein P 1 levels and glutathione peroxidase 3 activity were measured using an enzyme-linked immunosorbent assay. Ultra-performance liquid chromatography-tandem mass spectrometry was used to analyze proteinaceous Se species in enzymatically digested breast milk. RESULTS: Dietary Se intakes of lactating women from Liangshan, Beijing, and Enshi were 41.6±21.2 ng/d, 51.1±22.6 ng/d, and 615±178 ng/d, respectively (p<0.05). The Se levels in the blood and breast milk were significantly associated with the dietary Se intake (p<0.05). The proteinaceous Se species in breast milk were SeMet and SeCys2. The levels of SeMet in the lactating women from Liangshan, Beijing, and Enshi were 3.31±2.44 ng Se/mL, 7.34±3.70 ng Se/mL, and 8.99±9.64 ng Se/mL, while that of SeCys2 were 13.7±12.0 ng Se/mL, 35.6±20.9 ng Se/mL, and 57.4±13.2 ng Se/mL, respectively. Notably, the concentration of SeCys2, the metabolite of unstable SeCys, reached a saturation platform, whereas no similar phenomenon were found for the total Se SeMet from Secontaining proteins. CONCLUSIONS: SeCys2 in breast milk is a potential biomarker for determining the optimal Se intake in lactating women.

Calculation of an Adequate Intake (AI) Value and Safe Range of Selenium (Se) for Chinese Infants 0-3 Months Old Based on Se Concentration in the Milk of Lactating Chinese Women with Optimal Se Intake.

The required selenium intake for optimal health in Chinese residents was published in 2014. However, the adequate intake (AI) value for Chinese infants 0-3 months old is not established. This study assessed the current selenium nutritional status of 264 lactating Chinese women from three geographical locations with different Se levels (Liangshan in Sichuan province, Enshi in Hubei province, and Xicheng District in Beijing), to screen mothers with optimal Se intake, and to modify the AI value of Se for Chinese infants 0-3 months old. Milk and plasma Se concentrations were determined by inductively coupled plasma mass spectrometry (ICP-MS) and glutathione peroxidase 3 (GPx3), and plasma selenoprotein P (SEPP1) was measured by ELISA. Daily Se intake (Y, µg/day) in lactating Chinese woman was calculated from plasma Se concentrations (X, µg/L) using the formula logY = 1.623 log(X) + 3.433. Plasma Se concentrations in lactating Chinese women were 78.19 ± 25.71, 112.48 ± 24.57, and 183.83 ± 45.81 µg/L from Se-deficient, Se-moderate, and seleniferous areas, respectively. Se intakes calculated from concentrations of plasma Se were 45.6 ± 21.69, 80.03 ± 27.69, and 223.10 ± 50.95 µg/day, respectively. An optimal dietary Se intake is not lower than the recommended nutrient intake (RNI) but not more than the tolerable upper intake level (UL). A range of 78-400 µg Se/day was defined as the optimal daily Se intake for lactating Chinese women. The percentages of mothers within this range in Sichuan, Beijing, and Enshi were 8.11, 45.13, and 6.06%, respectively. Based on milk Se concentrations of mothers with optimal daily Se intake, the adequate Se intake value and a safe range for Chinese infants 0-3 months of age were calculated as 15.29 and 8-35 µg Se/day, respectively. The Se status of Chinese lactating women has improved, particularly in traditionally Se-deficient and Se-toxic regions. A safe range for daily Se intake in Chinese infants may be regarded as a guideline for infant formula.

Selenium in infant formula milk.

Formula-based animal milk is an alternative source of infant nutrition in many cases when breastfeeding is unacceptable or inaccessible; however, these replacements often have low selenium levels. The composition of infant formula milk should be as close as possible to that of human breast milk, both in content and chemical speciation. Selenium is an essential trace element for infants. Generally, human breast milk is the ideal food to ensure adequate infant Se intake. However, to date, sodium selenite or sodium selenate has been used as selenium supplementation in infant formula milk in most countries. This inorganic Se, which is not a natural component of food, may not be the optimal speciation for Se supplementation in infant formula milk. Advances in speciation in foods, especially in animal milk, suggest that future proposals for selenium speciation in human breast milk can lead to discussions regarding the most favorable methods of selenium supplementation in infant formula milk.

The Synergistic Effect of Serine with Selenocompounds on the Expression of SelP and GPx in HepG2 Cells.

We explored the synergistic effect of serine combined with several selenocompounds or used alone on the expression of selenoprotein P (SelP) and glutathione peroxidase (GPx) in this study. We first compared the SelP and GPx expression difference between HepG2 and Hela cells treated with serine and finally chose HepG2 as experimental cell. In the serine-used-alone experiment, three kinds of selenium nutritional models (low-, adequate-, and high-selenium) were established and serine was 10 times gradient diluted (0.01 to 100 µmol/L). In the combined experiment, the selenocompound doses were set as 0.01, 0.1, and 1 µmol Se/L and serine was set according to its molar ratio with the selenocompounds. We found that SelP and GPx concentrations in the low-, adequate-, and high-selenium models increased following with serine dose. When the concentration of sodium selenite and SeMet was 1 µmol Se/L while MeSeCys was 0.1 and 1 µmol Se/L, SelP concentrations for serine combined with selenocompounds groups were significantly higher than that of selenocompounds used alone. When the concentration of sodium selenite was 0.1 µmol Se/L, SeMet was 0.1 and 1 µmol Se/L while MeSeCys was 0.01 and 1 µmol Se/L, GPx concentrations for serine combined with selenocompounds groups were significantly higher than that of selenocompounds used alone. Our preliminary result indicated the beneficial effect of serine on the expression of SelP and GPx, which suggested that it might be a candidate for combined selenium supplement.

The effects of different levels of calcium supplementation on the bone mineral status of postpartum lactating Chinese women: a 12-month randomised, double-blinded, controlled trial.

Increasing dietary Ca intake may prevent the excessive mobilisation of bone mineral in nursing mothers. We aimed to investigate whether higher Ca intake could positively modulate the bone mineral changes in Chinese postpartum lactating women. The study was a 12-month randomised, double-blinded, parallel group trial conducted over 12 months. A total of 150 postpartum women were randomly selected to receive either 40 g of milk powder containing 300 mg of Ca and 5 µg of vitamin D (Low-Ca group) or same milk powder additionally fortified with 300 mg of Ca (Mid-Ca group) or 600 mg of Ca (High-Ca group). Bone mineral density (BMD) for the whole body, the lumbar spine, the total left hip and its sub-regions was measured using dual-energy X-ray absorptiometry. A total of 102 subjects completed the whole trial. The duration of total lactating time was 7·9 (SD 2·8) months on average. The intention-to-treat analysis yielded the following mean percentage changes in BMD for the whole body, the lumbar spine and the total left hip, respectively: -0·93 (SD 1·97), 2·11 (SD 4·90) and -1·60 (SD 2·65)% for the Low-Ca group; -0·56 (SD 1·89), 2·21 (SD 3·77) and -1·43 (SD 2·30)% for the Mid-Ca group; and -0·44 (SD 1·67), 2·32 (SD 4·66) and -0·95 (SD 4·08)% for the High-Ca group. The differences between the groups were not statistically significant (P: 0·5-0·9). The results of the complete case analysis were similar. In sum, we found no significant differences in the bone mineral changes from baseline to 12 months in postpartum lactating women consuming milk powder fortified with different levels of Ca.