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Obesity, caused by an increased number and volume of adipocytes, is a global epidemic that seriously threatens human health. Bone marrow mesenchymal stem cells (BMSCs) can differentiate into adipocytes. All-trans retinoic acid (atRA, the active form of vitamin A) inhibits the adipogenic differentiation of BMSCs through its receptor RARG. The expression level of FRA1 (FOS like 1, AP-1 transcription factor subunit) in atRA-treated BMSCs increased, suggesting that atRA-mediated inhibition of BMSCs adipogenesis involves FRA1. BMSCs were transfected with adenovirus overexpressing Fra1 (ad-fra1) or silenced for Fra1 (si-fra1) and then treated with atRA. BMSCs treated with atRA and treated with ad-fra1 showed decreased mRNA and protein levels of key adipogenic genes (Pparg2, Cebpa) and adipogenesis-associated genes (Cd36, Fabp, Lpl, and Plin); atRA had a stronger inhibitory effect on adipogenesis compared with that in the ad-fra1 group. Adipogenic gene expression in Fra1-silenced BMSCs was significantly upregulated. Compared with that in the atRA group, the si-fra1 + atRA also upregulated adipogenic gene expression. However, compared with si-fra1, si-fra1 + atRA significantly inhibited adipogenic differentiation. Chromatin immunoprecipitation showed that RARG directly regulates Fra1 and FRA1 directly regulates Pparg2 and Cebpa. The results supported the conclusion that atRA inhibits BMSC adipogenesis partially through the RARG-FRA1-PPARG2 or the CEBPA axis or both. Thus, vitamin A might be used to treat obesity and its related diseases.

Our previous studies demonstrated that vitamin A deficiency (VAD) can impair the postnatal cognitive function of rats by damaging the hippocampus. The present study examined the effects of retinoic acid (RA) on apoptosis induced by hypoxic-ischemic damage in vivo and in vitro, and investigated the possible signaling pathway involved in the neuroprotective anti-apoptotic effects of RA. Flow cytometry, immunofluorescence staining and behavioral tests were used to evaluate the neuroprotective and anti-apoptotic effects of RA. The protein and mRNA levels of RARα, PI3K, Akt, Bad, caspase-3, caspase-8, Bcl-2, Bax, and Bid were measured with western blotting and real-time PCR, respectively. We found impairments in learning and spatial memory in VAD group compared with vitamin A normal (VAN) and vitamin A supplemented (VAS) group. Additionally, we showed that hippocampal apoptosis was weaker in the VAN group than that in VAD group. Relative to the VAD group, the VAN group also had increased mRNA and protein levels of RARα and PI3K, and upregulated phosphorylated Akt/Bad levels in vivo. In vitro, excessively low or high RA signaling promoted apoptosis. Furthermore, the effects on apoptosis involved the mitochondrial membrane potential (MMP). These data support the idea that sustained VAD following hypoxic-ischemic brain damage (HIBD) inhibits RARα, which downregulates the PI3K/Akt/Bad and Bcl-2/Bax pathways and upregulates the caspase-8/Bid pathway to influence the MMP, ultimately producing deficits in learning and spatial memory in adolescence. This suggests that clinical interventions for HIBD should include suitable doses of VA. Electronic supplementary material The online version of this article (10.1186/s13041-018-0360-0) contains supplementary material, which is available to authorized users.

Hypoxia-ischemia (HI)-induced perinatal encephalopathy frequently causes chronic neurological morbidities and acute mortality. Bone mesenchymal stem cell (BMSC) transplantation could potentially promote functional and anatomical recovery of ischemic tissue. In vitro hypoxic preconditioning is an effective strategy to improve the survival of BMSCs in ischemic tissue. In this study, cobalt chloride (CoCl2) preconditioned medium from BMSC cultures was injected into the left lateral ventricle of HI rats using a micro-osmotic pump at a flow rate 1.0μl/h for 7 days. The protein levels of HIF-1α and its target genes, vascular endothelial growth factor and erythropoietin, markedly increased after CoCl2 preconditioning in BMSCs. In 7-week-old rats that received CoCl2 preconditioned BMSC medium, results of the Morris water maze test indicated ameliorated spatial working memory function following hypoxia-ischemia damage. Neuronal loss, cellular disorganization, and shrinkage in brain tissue were also ameliorated. Extracellular field excitatory postsynaptic potentials (fEPSPs) in the brain slices of 8-week-old rats were recorded; administration of CoCl2 preconditioned BMSC culture medium induced a progressive increment of baseline and amplitude of the fEPSPs. Immunohistochemical quantification showed that GluR2 protein expression increased. In conclusion, CoCl2 activates HIF-1α signals in BMSCs. CoCl2 preconditioned BMSC culture medium likely effects neuroprotection by inducing long-term potentiation (LTP), which could be associated with GluR2 expression. The paracrine effects of hypoxia preconditioning on BMSCs could have applications in novel cell-based therapeutic strategies for hypoxic and ischemic brain injury.

Background Although clinical vitamin A deficiency (VAD), which is a public health problem developing throughout the world, has been well controlled, marginal vitamin A deficiency (MVAD) is far more prevalent, especially among pregnant women and preschool children in China. Increasing evidence suggests that VAD is involved in the pathogenesis of Alzheimer's disease (AD). However, whether MVAD, beginning early in life, increases the risk of developing AD has yet to be determined. Objective The goal of this study was to investigate the long-term effects of MVAD on the pathogenesis of AD in rats. Method An MVAD model was generated from maternal MVAD rats and maintained with an MVAD diet after weaning. The males were bilaterally injected with aggregated amyloid β (Aβ)1-42 into the CA3 area of the hippocampus, and the AD-associated cognitive and neuropathological phenotypes were examined. Results We found that MVAD feeding significantly aggravated Aβ1-42-induced learning and memory deficits in the Morris water maze test. MVAD did not induce the mRNA expression of retinoic acid receptors (RARs), a disintegrin and metalloprotease 10 (ADAM10) or insulin-degrading enzyme (IDE) in Aβ1-42-injected rats. Moreover, RARα and RARγ mRNA were positively correlated with ADAM10 mRNA, whereas RARβ mRNA was positively correlated with IDE mRNA. Conclusion Our study suggests that MVAD beginning from the embryonic period perturbs the ADassociated genes, resulting in an enhanced risk of developing AD.

Maternal folate and vitamin B12 deficiency predict poor pregnancy outcome. To improve pregnancy outcomes in rural area of China, we investigate rural women's folic acid supplementation (FAS) status and the associations between maternal vitamin B status during the first trimester and subsequent adverse pregnancy outcomes. We collected the questionnaire information and drew 5 ml of blood from 309 early pregnant rural women. The birth outcomes were retrieved from medical records after delivery. Out of the total, 257 had taken FAS, including 50 before conception (group A) and 207 during the first trimester (group B). The concentration of plasma folate and the RBC folate supplementation groups were obviously higher than that of no-supplementation group (group N, p

Vitamin A (VA) is an essential micronutrient. Numerous studies have confirmed that VA deficiency (VAD) leads to a decline in learning and memory function. Our previous studies have demonstrated that retinoic acid nuclear receptor α (RARα) in the hippocampus plays a crucial role in learning and memory, but the exact mechanism for this process is unclear. Epigenetic modifications, particularly histone acetylation, are involved in nervous system development, learning and memory function, and the pathogenesis of neurodegenerative diseases. Histone acetyltransferases (HATs), such as CREB-binding protein (CBP), E1A-binding protein p300 (p300), and p300/CBP-associated factor (PCAF), are critical for regulating memory function. The current study uses RARα and CBP as examples to study the connections between the RA signaling pathway and histone acetylation modification and to reveal the epigenetic mechanism in VAD-induced learning and memory impairment. This study examined the expression of RARα, HATs, acetylated histone H3/H4, and memory-related genes (Zif268, cFos, FosB), as well as the interaction of RARα and CBP in the hippocampus of 8-week-old rats. Additionally, the changes shown in vivo were further assessed in primary cultured neurons with the inhibition or overexpression of RARα. We found significantly lower levels of histone acetylation in the VAD rats. Furthermore, this downregulation, which impairs learning and memory, is induced by the dysregulation of CBP-dependent histone acetylation that is mediated by RARα. This work provides a solid theoretical foundation and experimental basis for the importance of ensuring sufficient nutritional VA during pregnancy and early life to prevent impairments of learning and memory in adulthood.

Background: Hypoxia–ischemia (HI)-induced perinatal encephalopathy is a major cause of acute mortality and chronic neurologic morbidities such as cerebral palsy, mental retardation, and epilepsy. As the essential transcription factor for the activation of hypoxia-inducible genes, hypoxia-inducible factor 1 alpha (HIF-1α) plays an important role in the pathophysiological response to the stress of HI brain damage. Whether HIF-1α activation promotes neuroprotection in HI tissues is controversial. Methods: The left common carotid artery of rats aged 7days was ligated under anesthesia. The pups were then exposed to hypoxia in a normobaric chamber filled with 8% oxygen and 92% nitrogen for 2.5h. In the sham control group, the left common carotid artery was exposed but was not ligated or exposed to hypoxia. To assess the time window for effective treatment, the HIF-1α inducer cobalt chloride (CoCl 2 ) was injected subcutaneously 1day before surgery, immediately or 1day after surgery. The brain tissues were harvested from the pups of each groups at 1, 2 and 7days after insult for HIF-1α protein ant its target genes expression and for investigating the injury. Morris water maze tests were performed at postnatal 7weeks. Results: HIF-1α protein levels and its target genes vascular endothelial growth factor, heme oxygenase-1, and insulin-like growth factor 1 were markedly increased after intraperitoneal injection of CoCl 2 (60mg/kg). The target gene inducible nitric oxide synthase exhibited a biphasic time course. HI caused apoptosis and reduced capillary density, which were ameliorated by CoCl 2 . Both preconditioning with CoCl 2 24h before HI and administration of CoCl 2 24h after HI improved long-term reference memory compared with that in vehicle-injected littermate controls. Administration of CoCl 2 immediately after HI did not improve spatial working memory. Conclusions: CoCl 2 activates HIF-1α and protects against brain damage in vivo. The time of administration could be used to manipulate the activity of HIF-1α pathways and promote recovery.

Background: Histone acetylation, which is a chromatin modification of histone tails, can dynamically regulate the expression of various genes in normal development. HDAC2 is a negative regulatory factor of acetylation and closely related to learning and memory. NSE is a nerve marker and vital for maintaining physiological functions in nervous system. Currently, few studies associated with the expression pattern of HDAC2 in postnatal rat hippocampus have been reported. This study aimed to explore the temporal and spatial expression pattern of HDAC2, helping to reveal the expression characteristics of HDAC2 during postnatal neuronal maturation. Materials and Methods: With NSE as a biomarker of neuronal maturation at postnatal days 1, 3, 7 and weeks 2, 4, and 8 (P1D, P3D, P7D, P2W, P4W, P8W), the expression patterns of HDAC2 in rat hippocampus were examined using real-time PCR and western blotting. Additionally, the subcellular distribution of HDAC2 was analysed by immunofluorescence. Results: We found that HDAC2 was highly expressed in the neonatal period and decreased gradually. HDAC2 expression was widely distributed in neurons of hippocampal CA1, CA3 and DG regions and gradually shifted from the nucleus to the cytoplasm during postnatal development. Altogether, the expression of HDAC2 decreased gradually with different subcellular localizations throughout development. Conclusions: The observed results indicate that the expression levels of HDAC2 become lower and with different subcellular localizations in neurons during hippocampal neuronal maturation, suggesting the specific expression characteristics of HDAC2 might play an important role during postnatal learning-memory function and development.

Pre-activation of the retinoid signaling pathway by all-trans retinoic acid facilitates neuronal differentiation of mesenchymal stem cells. Using protein/DNA based screening assays, we identified activator protein 2α as an important downstream target of all-trans retinoic acid. Although all-trans retinoic acid treatment significantly increased activator protein 2α transcriptional activity, it did not affect its expression. Inhibition of activator protein 2α with dominant-negative mutants reduced ATRA-induced differentiation of mesenchymal stem cells into neurons and reversed its associated functional recovery of memory impairment in the cell-based treatment of a hypoxic-ischemic brain damage rat model. Dominant-negative mutants of activator protein 2α inhibited the expression of neuronal markers which were induced by retinoic acid receptor β activation. All-trans retinoic acid treatment increased phosphorylation of activator protein 2α and resulted in its nuclear translocation. This was blocked by siRNA-mediated knockdown of retinoic acid receptor β. Furthermore, we found that retinoic acid receptor β directly interacted with activator protein 2α. In summary, the regulation of all-trans retinoic acid on activator protein 2α transcriptional activity was mediated by activation of retinoic acid receptor β and subsequent phosphorylation and nuclear translocation of activator protein 2α. Our results strongly suggest that activator protein 2α transcriptional activity is essential for all-trans retinoic acid-induced neuronal differentiation of mesenchymal stem cells.

The aim of our study was to examine the association of vitamin A status with obesity and the metabolic syndrome (MS) in school-age children in Chongqing, China. A cross-sectional study was conducted of 1,928 children aged 7~11 years from 5 schools in Chongqing, China. Body height, weight, waist circumference (WC) and blood pressure were measured. Blood glucose, lipids and vitamin A were determined. Overall prevalences for overweight, obesity and MS were 10.1%, 6.7% and 3.5%, respectively. There were 274 (14.2%) marginally vitamin A deficient (MVAD) children and 53 (2.8%) vitamin A deficient (VAD) children, respectively. Serum vitamin A in the obese group was significantly lower than in the overweight and normal weight groups (p0.001). Body mass index (BMI), WC, high density lipoprotein cholesterol (HDL-C) and glucose were strongly associated with vitamin A status (p0.05). In a separate model adjusted for age and sex, compared with normal children, participants with obesity had a significantly higher risk of having vitamin A insufficiency (=1.05 μmol/L) (OR: 2.37; 95% CI: 1.59, 3.55) (p0.001), and participants with MS had a 1.99-fold (95% CI: 1.14, 3.47) greater risk of having vitamin A insufficiency (p=0.016), while participants with VAD had significantly higher risk of having MS (OR: 3.82; 95% CI: 1.44, 10.2) (p=0.007). Vitamin A insufficiency among Chongqing urban school-age children was found to be a severe health problem, significantly associated with obesity, hypertriglyceridemia and MS.本研究旨在观察中国重庆地区学龄期儿童维生素A 营养状况与肥胖和代谢综 合征的关系。横断面调查重庆市渝中区5 所小学共1,928 名学龄期儿童。检查 其身高、体重、腰围和血压水平，检测其血清维生素A、血糖和血脂水平。结 果显示，调查对象的超重率为10.1%，肥胖率为6.7%；代谢综合征发生率为 3.5%；维生素A 缺乏（VAD）发生率为2.8%，边缘型维生素A 缺乏 （MVAD）发生率为14.2%。肥胖组儿童血清维生素A 水平显著低于超重组 和正常组。体质指数、腰围、高密度脂蛋白胆固醇和血糖水平与维生素A 营 养状况有关（p0.05）。校正年龄和性别后，肥胖儿童发生维生素A 不足 （=1.05 μmol/L）的风险是正常儿童的2.37 倍（95% CI：1.59，3.55) ， （p0.001），代谢综合征儿童发生维生素A 不足的风险是正常儿童的1.99 倍 （95% CI：1.14，3.47，p=0.016），同时我们发现，VAD 的儿童发生代谢综 合征的风险是正常儿童的3.82 倍（95% CI：1.44，10.2，p=0.007）。重庆市 城区学龄期儿童VA 不足的发生率较高，是一个较严重的健康问题，维生素A 不足与肥胖和代谢综合征有关。.

This study aims to construct the recombinant lentivirus vector containing specific small interfering RNA (siRNA) targeting rat CREB binding protein(CBP)gene and to identify its function of inhibiting the expressions of acetylated histone in primarily cultured hippocampal neurons. Firstly, we constructed four kinds of recombinant lentivirus siCBP. And then we used them to infect the primarily cultured hippocampal neurons, and performed real-time PCR, western blot respectively to detect the expressions of CBP. Afterwards, the most effective lentivirus siCBP was used to infect the primarily cultured hippocampal neurons, and then the HAT activity and protein expressions of acetylated histone Ac-H3, Ac-H4 of the neurons were examined. By using PCR, endonuclease cutting and gene sequencing, we confirmed that the target genes were correctly cloned in lentivirus vector. Besides, CBP mRNA and protein expressions in neurons were found to be with varying degrees of decreases after infections of the four kinds of lentivirus siCBP. Furthermore, the representative and most effective lentivirus GR806 could effectively inhibit the HAT activity and the protein expressions of Ac-H3, Ac-H4 in neurons. It provides the experimental basis for the subsequent application of siCBP to clarify the effects and corresponding molecular mechanism of the CBP-dependent histone acetylation on learning and memory function in hippocampus.

Mesenchymal stem cells (MSCs) can differentiate into neurons in an appropriate cellular environment. Retinoid signaling pathway is required in neural development. However, the effect and mechanism through retinoid signaling regulates neuronal differentiation of MSCs are still poorly understood. Here, we report that all-transretinoic acid (ATRA) pre-induction improved neuronal differentiation of rat MSCs. We found that, when MSCs were exposed to different concentrations of ATRA (0.01–100 lmol ⁄ L) for 24 h and then cultured with modified neuronal induction medium (MNM), 1 lmol ⁄ L ATRA pre-induction significantly improved neuronal differentiation efficiency and neural-cell survival. Compared with MNM alone induced neural-like cells, ATRA ⁄ MNM induced cells expressed higher levels of Nestin, neuron specific enolase (NSE), microtubule-associated protein-2 (MAP-2), but lower levels of CD68, glial fibrillary acidic protein (GFAP), and glial cell line-derived neurotrophic factor(GDNF), also exhibited higher resting membrane potential and intracellular calcium concentration, supporting that ATRA pre-induction promotes maturation and function of derived neurons but not neuroglia cells from MSCs. Endogenous retinoid X receptors (RXR) RXRa and RXRc (and to a lesser extent, RXRb) were weakly expressed in MSCs. But the expression of RARa and RARc was readily detectable, whereas RARb was undetectable. However, at 24 h after ATRA treatment, the expression of RARb, not RARa or RARc, increased significantly. We further found the subnuclear redistribution of RARb in differentiated neurons, suggesting that RARb may function as a major mediator of retinoid signaling during neuronal differentiation from MSCs. ATRA treatment upregulated the expression of Vimentin and Stra13, while it downregulated the expression of Brachyury in MSCs. Thus, our results demonstrate that pre-activation of retinoid signaling by ATRA facilitates neuronal differentiation of MSCs.

ObjectiveTo investigate the efficacy of biscuits fortified with different doses of vitamin A on improving vitamin A deficiency (VAD), anaemia and physical growth of pre-school children.DesignA randomised double-masked population-based field interventional trial with a positive control group.SettingBanan district of Chongqing, China.SubjectsA total of 580 pre-school children aged 3–6 years were randomly recruited into four groups. Children in groups I and II were given biscuits fortified with vitamin A at 30 % of the recommended daily intake (RDA) and 100 % of the RDA once a day for 9 and 3 months, respectively. Children in group III received biscuits containing 20 000 IU of vitamin A once a week for 3 months. Initially, the children in group IV received a 200 000 IU vitamin A capsule just once. At the beginning and end of the study, blood samples were collected to measure Hb, serum retinol, retinol-binding protein and prealbumin, and weight and height were measured.ResultsAll the fortification types significantly decreased the prevalence of VAD and anaemia in each group (P < 0·05). The effect of 9-month intervention on group I was the most efficient (P < 0·0045). After intervention, the Z-scores of height-for-age, weight-for-age and weight-for-height in all groups increased markedly compared with baseline (P < 0·05), but no significant difference was observed among the groups.ConclusionsData indicated that consuming vitamin A-fortified biscuits with daily 100 % RDA for 3 months has the same effect on the improvement of VAD, anaemia and physical growth as did the weekly 20 000 IU and single 200 000 IU administration in pre-school children.

Anemia is a widespread public health problem, which is due to many factors, nutritional or non-nutritional. Iron, vitamin A and growth status were assessed to investigate anemia of preschool children in suburb Chongqing, China. A descriptive, cross-sectional survey was performed on 459 preschool children aged 2 to 7 years randomly chosen from the kindergartens in 6 suburban districts of Chongqing. Weight and height levels, hemoglobin, erythrocyte protoporphyrin, serum retinol, and ferritin concentrations were measured to evaluate the anthropometric and nutritional status. The rates of stunt, underweight, overweight, wasting, obesity, anemia, iron deficiency, vitamin A deficiency (VAD), and marginal VAD were 6.3%, 3.9%, 3.7%, 1.5%, 3.1%, 23.5%, 15.0%, 6.3% and 25.9%, respectively. Serum retinol concentration was significantly lower in children with anemia than in those without anemia (P=0.003), and the retinol concentration was associated with hemoglobin (Pearson’s correlation coefficient, r=0.22, P

To investigate the correlation of the antioxidant vitamins status (vitamins A, E and C) during pregnancy and the intellectual development of early childhood.A total of 150 paired maternal-neonatal subjects were recruited into the present study. The serum concentrations of antioxidant vitamins (vitamins A, E and C) in maternal blood and cord blood after delivery were determined by high performance liquid chromatography and the intellectual development was evaluated by Gesell Development Schedules (GDS) at two-years-old.Children with higher cord serum vitamin E level showed higher scores of motor, adaptive domain and average compared to children with lower cord serum vitamin E level (p0.01 or 0.05), respectively. Cord serum vitamin A level had significant positive correlation with effect on motor DQs (beta=4.227, p0.05), and vitamin E level in cord blood showed a positive relation with motor DQ and average DQ (beta=0.329 and 0.1875, respectively, p0.05) in multiple linear regression model. The language and social DQs were influenced by placental vitamin E transport rate (beta=3.1968 and 3.0194, respectively, p0.05). The placental transport rate of vitamin E also was a protective factor for the prevalence of motor behavior developmental delay [OR: 0.118, 95% confident interval (95% CI), 0.018-0.765, p=0.0251], personal and social behavior developmental delay (OR: 0.052, 95% CI: 0.004-0.610, p=0.0185) and average developmental delay (OR: 0.041, 95% CI: 0.003-0.642, p=0.0229) in logistic multiple regression model.Data suggested that vitamin A, E status and vitamin E transfer rate at delivery had beneficial influence on children's cognitive and behavior development quotients.

Mild or marginal vitamin A deficiency (MVAD) is still a serious and widespread public health problem in pregnant women and children in developing countries. This study investigated rat lung maturation from prenatal to adult stage during pregnancy and postnatal MVAD and the recovery after postnatal vitamin A supplementation (VAS). Adult female rats and their offspring were randomized into three groups. 1. Control: the mothers and offspring received a normal diet.2. MVAD: The mothers and offspring received a MVAD diet.3. VAS: the mothers received MVAD diet till parturition, and then received the normal diet. The offspring of the VAS group were given low-dose vitamin A from postnatal day 1 to day 7 and received the normal diet after weaning. The lung development, structure, and collagen and elastic fiber of offspring were monitored by morphometric analysis at age 1 d, 2 and 8 wk, respectively. Lower body weight, lung weight, reduced numbers of alveoli and total alveolar surface area as well as increased alveoli septa thickness was observed in MVAD compared to that in the control animals. Increased collagen deposits and decreasing elastic fiber were found in MVAD rats. However, all of these were significantly improved in VAS-treated animals. These data suggest that the rat lung is sensitive to MVAD during the developing stage. Early postnatal vitamin A supplementation can partially restore the normal lung structure.

Retinoic acid receptors (RARs) are reported to mediate the effects of retinoid acid and participate in the maintenance of normal hippocampal function during embryonic and postnatal stages. RARalpha is the only one that has been reported to be continuously expressed among RARs in the CA1-CA3 areas of the hippocampus, at both the mRNA and the protein level. Here, we show the expression and subcellular localization of RARalpha in granule and pyramidal cells in various regions of the hippocampus during postnatal development of rats. We discovered that the expression level of RARalpha in postnatal hippocampal tissue gradually decreased over time with increasing developmental maturity of the nervous system. Moreover, the subcellular localization of RARalpha expression showed a phenomenon of intracellular translocation during the postnatal development period. This new discovery is inconsistent with a traditional viewpoint according to which RARalpha, as a nuclear transcription factor, is mainly expressed inside nucleus. This phenomenon suggests that RARalpha may have different actions during each stage of hippocampal development.

Preschool children in developing countries are likely to have multiple, concurrent micronutrient deficiencies. This study was designed to evaluate the effectiveness of different combinations of nutritional fortified diet to improve the blood levels of iron, vitamin A and other essential micronutrients in the preschool population of Banan District of Chongqing, China. From December 2005 to June 2006, a total of 226 2-6 y old preschool children were recruited from three nurseries in the area, and they were randomly assigned to three different fortified diet groups for 6 mo. Group I was fortified with vitamin A; groups II and III were fortified with vitamin A plus iron and vitamin A plus iron, thiamine, riboflavin, folic acid, niacinamide, zinc and calcium, respectively. Subjects' weight and height were measured for assessing the children's growth and development. Blood samples were taken at the beginning and the end of the 6-mo study period for measuring serum levels of micronutrients. Group III with the multiple micronutrient fortified diet was the most effective to improve the serum level of retinol from [media (P25, P75): 1.06 (0.89, 1.32)] micromol/L to 1.29 (1.04, 1.39) micromol/L (p0.05) and retinol binding protein from 17.0 (12.6, 25.6) mg/L to 31.6 (24.4, 44.0) mg/L (p0.05) and to mobilize the stored iron in the liver (p0.05). In addition, the three groups' hemoglobin levels were elevated from 117.0 (109.0, 124.1) g/L, 114.0 (109.2, 119.7) g/L and 115.0 (109.5, 122.7) g/L to 125.7 (119.2, 133.1) g/L, 126.5 (122.2, 135.9) g/L and 125.1 (119.8, 131.6) g/L over the 6 mo of intervention period, but there were no difference among the three groups (p0.05). Nevertheless, unexpected results were obtained when comparing the effects on growth status among the different supplement groups. Our study has demonstrated that a multiple micronutrient fortified diet for 6 mo is more effective to improve the levels of hemoglobin, serum retinol, and RBP as well as to facilitate the mobilization of iron storage in preschool children.

This study analyzed the effects of short hairpin RNA targeting interleukin-1 receptor-associated kinase-4 (IRAK-4-shRNA) via portal vein perfusion during different periods on ischemia/reperfusion injury after liver transplantation.Rats were randomly divided into 3 groups: the cold ischemia transfection group (CIT group, n = 18), in which graft livers were perfused with the plasmid of pSIIRAK-4 expressing IRAK-4-shRNA for 4 minutes (0.5 mL/min) via the portal vein during the cold ischemia period; the in vivo transfection group (IVT group, n = 18), in which equivalent volumes (2 mL) of IRAK-4-ShRNA plasmid (pSIIRAK-4) were injected during the operation; and the control group (n = 18), in which the rats received equivalent volumes of blank plasmid. At 0, 60, and 180 minutes after portal vein reperfusion, blood and liver tissues were collected for examination. IRAK-4 expression, nuclear factor kB (NF-kB) activity, tumor necrosis factor α, interleukin (IL)-1β, and IL-6 serum levels, as well as histologic changes, were detected.At 0 minutes after reperfusion, IRAK-4 expression, NF-κB activity, and serum levels of tumor necrosis factor α, IL-1β, and IL-6 showed no significant differences among the 3 groups (P.05). At 60 and 180 minutes after reperfusion, all indices of the IVT and control groups were significantly higher than those of the CIT group (P .01). Meanwhile, all indices of the CIT group showed no significant differences at various time points (P.05). Liver function and histologic changes exhibited less liver injury in the CIT group than in the other groups.IRAK-4 activity was suppressed by IRAK-4-shRNA through portal vein perfusion during the cold ischemia period, and IRAK-4-shRNA effectively prevented ischemia/reperfusion injury after liver transplantation.

Purpose Because the choice of anesthetic affects the rodent orthotopic liver transplantation (OLT) model, we compared the effects of isoflurane, ketamine, chloral hydrate, and pentobarbital on the OLT model. Basic Procedures OLT was performed using the two-cuff technique. Two hundred male rats were randomly divided into five groups: control, isoflurane, ketamine, chloral hydrate, and pentobarbital groups. Rectal temperatures, respiratory rates, arterial blood values (pH, PaCO2, PaO2, and SatO2), liver function tests and histopathology, recovery times, and anhepatic stage mortality rates were assessed. Main Findings Compared with controls, respiratory rates decreased by 20% in the isoflurane group, and decreased by 40%-50% in the ketamine, chloral hydrate, and pentobarbital groups. The PaO2, SatO2, and pH levels in the ketamine, chloral hydrate, and pentobarbital groups were significantly lower than those in the isoflurane and control groups (P Principal Conclusions Isoflurane should be the preferred anesthetic for rodent OLT surgery due to its minimal respiratory and hepatic physiological effects as well as its low anhepatic phase mortality rate. Secondary to isoflurane, ketamine and chloral hydrate may be administered as donor anesthetics. Pentobarbital use should be avoided entirely in rodent OLT surgery due to its significant hepatotoxic effects.

Objectives Oxidative stress (OS) may play a critical role in cell aging and neurologic disorders that are often seen in Down syndrome (DS) patients. The aim of this study was to determine the antioxidant enzyme level and lipoperoxidation status in blood from DS children. Design and methods In a cross-sectional study, we recruited a total of 36 DS children and 40 healthy controls (HCs). All subjects were free of infection according to the C reactive protein (CRP) value and routine peripheral blood profile. The activities of total superoxide dismutases (SODs), extracellular glutathione peroxidase (GPx3),malondialdehyde (MDA) and nitric oxide synthase (NOS) concentrations in peripheral blood were measured by spectrophotometric methods. The relationship of SOD and GPx3 was analyzed in the two groups. Results The two groups were similar with respect to age, gender and peripheral blood profiles. The total SOD activity was significantly increased, while the GPx3 activity was significantly reduced in the DS group compared to the HCs (p = 0.000, p = 0.033 respectively). The MDA level was higher in DS children (p = 0.013). There was no significant difference in NOS between DS and HCs (p = 0.708). A significant negative correlation between GPx3 and SOD activity was identified in DS (r = − 0.14, p = 0.018) but not in the HC group. Conclusions Abnormal redox metabolism takes place in DS individuals. Reducing GPx3 may be a compensatory mechanism of protection against intracellular OS. Moreover, monitoring of decreases in GPx3 activity may be a useful biomarker for evaluating OS in DS patients.

Background Alcohol is detrimental to early development. Fetal alcohol spectrum disorders (FASD) due to maternal alcohol abuse results in a series of developmental abnormalities including cranial facial dysmorphology, ocular anomalies, congenital heart defects, microcephaly and intellectual disabilities. Previous studies have been shown that ethanol exposure causes neural crest (NC) apoptosis and perturbation of neural crest migration. However, the underlying mechanism remains elusive. In this report we investigated the fetal effect of alcohol on the process of neural crest development in the Xenopus leavis. Results Pre-gastrulation exposure of 2-4% alcohol induces apoptosis in Xenopus embryo whereas 1% alcohol specifically impairs neural crest migration without observing discernible apoptosis. Additionally, 1% alcohol treatment considerably increased the phenotype of small head (43.4% ± 4.4%, total embryo n = 234), and 1.5% and 2.0% dramatically augment the deformation to 81.2% ± 6.5% (n = 205) and 91.6% ± 3.0% (n = 235), respectively (P

Background Non-communicable diseases are leading causes of disease burden in middle income countries. Little evidence exists to determine if the primary healthcare system can effectively manage non-communicable diseases. The purpose of this study was to examine the effectiveness of hypertension and diabetes management by the primary healthcare system. Methods We used individual level data from the 2009 National Basic Public Health Services System to assess the effectiveness of hypertension and diabetes interventions on fasting plasma glucose, and blood pressure. We analyzed the associations between fasting plasma glucose, systolic or diastolic blood pressure and risk factors. The estimated average intervention effect on data balanced with confounding variables was assessed. Results 9543 individuals who had data for fasting plasma glucose, systolic blood pressure and diastolic blood pressure were included in this analysis. This study included 6681 patients with hypertension and 2222 with diabetes. The intervention lowered mean fasting plasma glucose by 0.5 mmol/L (0.4–0.6), lowered mean systolic blood pressure by 3.5 mm Hg (3.2–3.7), and lowered diastolic blood pressure by 2.9 mm Hg (2.7–3.2). Individuals who received medicinal treatment had 1.3 mmHg (0.8 to 1.8, P

Lack of protein and vitamin A influences the growth of student in impoverished mountain areas. The aim of the study was to assess the effects of egg and vitamin A supplementation on hemoglobin, serum retinol and anthropometric indices of 10-18 years old students of a low socioeconomic status. A total number of 288 students from four boarding schools were randomly selected by using cluster sampling method in Chongqing, and they were assigned into supplement group and control group non-randomly. Students in supplement group received a single 200,000 international units vitamin A and 1 egg/day (including weekends) for 6 months. The control group did not receive any supplementation. We measured hemoglobin, serum retinol and height and weight at baseline and after supplementation. The supplementation increased the mean hemoglobin concentration by 7.13 g/L compared with 1.38 g/L in control group (p0.001), the mean serum retinol concentration by 0.31 μmol/L compared with 0.09 μmol/L in the control group (p=0.005), the mean height-for-age z score by 0.05 compared with 0.03 in the control group (p=0.319), the mean weight-for-age z score by 0.05 compared with -0.12 in the control group (p0.001). Our results revealed that egg and vitamin A supplementation is an effective, convenient, and practical method to improve the levels of hemoglobin, serum retinol and prevent the deterioration of growth in terms of weight for primary and middle school students from outlying poverty-stricken areas. Our intervention did not have a beneficial effect on linear growth.

Objective The aim of this study was to investigate the role of invariant natural killer T (iNKT) cells in liver transplant tolerance in rats. Methods Animals were randomly divided into 3 groups. The α-galactosylceramide (α-GalCer) group underwent injection through the caudal vein; the saline group received the same dose of saline and the control group received no treatment. Ten rats in each group were examined for survival the others were humanely killed on the seventh day posttransplantation. Liver tissues were used to assess histopathologic changes. Real-time polymerase chain reaction (PCR) was performed to determine the relative expressions of messanger RNAs of Th1/Th2-related cytokine (mRNAs) in the liver allograft. The serum levels of related cytokines were determined using enzyme-linked immunosorbent assay (ELISA). Results Allograft survival was significantly higher among the α-galactosylceramide α-GalCer group than the saline or control groups. The histopathology showed mild changes in the α-galactosylceramide α-GalCer compared with the other 2 groups. Real-time PCR showed the relative expression of Th1-related cytokine interferen (IFN)-γ mRNA to be significantly lower in the α-galactosylceramide α-GalCer compared with the other 2 groups, while the Th2-related cytokine interleukin (IL)-10 mRNA was much higher. The ELISA results confirmed these differential expressions. Conclusion The iNKT cells may play a pivotal role in liver transplant tolerance due to their regulatory functions on the Th1/Th2 imbalance. iNKT cells should be considered to be significant targets because of their attractive specificity and induction of liver allograft tolerance.

Aims To study the dynamic changes and the immunologic role of indoleamine 2, 3-dioxygenase (IDO) in Kupffer cells (KCs) after rat liver transplantation. Methods Animals were randomly divided into two groups: a rejection group (REJ; LEW to BN) and a tolerance group (TOL; BN to LEW). Liver morphological changes were observed optically with hematoxylin/eosin staining. KCs were isolated from recipients. mRNA and protein expressions of IDO were detected by real-time polymerase chain reaction and Western blotting at 1, 3, 5, and 7 days after transplantation. Results The levels of IDO mRNA and protein in KCs of TOL groups were similar to those in REJ groups at day 1 posttransplantation. However, the expression of IDO mRNA and protein time-dependently increased to much higher levels in the TOL than the in REJ groups at 3, 5, and 7 days posttransplantation ( P Conclusions The IDO level of KCs was closely associated with immune tolerance induction. IDO-mediated immune modulation appears to be an attractive means to assess transplant tolerance induction.

Background Mesenchymal stem cells (MSCs) can be induced to differentiate into neuronal cells under appropriate cellular conditions and transplanted in brain injury and neurodegenerative diseases animal models for neuroregeneration studies. In contrast to the embryonic stem cells (ESCs), MSCs are easily subject to aging and senescence because of their finite ability of self-renewal. MSCs senescence seriously affected theirs application prospects as a promising tool for cell-based regenerative medicine and tissue engineering. In the present study, we established a reversible immortalized mesenchymal stem cells (IMSCs) line by using SSR#69 retrovirus expressing simian virus 40 large T (SV40T) antigen as an alternative to primary MSCs. Methods The retroviral vector SSR#69 expressing simian virus 40 large T (SV40T) antigen was used to construct IMSCs. IMSCs were identified by flow cytometry to detect cell surface makers. To investigate proliferation and differentiation potential of IMSCs, cell growth curve determination and mesodermal trilineage differentiation tests were performed. Neuronal differentiation characteristics of IMSCs were detected in vitro. Before IMSCs transplantation, we excluded its tumorigenicity in nude mice firstly. The Morris water maze tests and shuttle box tests were performed five weeks after HIBD models received cells transplantation therapy. Results In this study, reversible IMSCs were constructed successfully and had the similar morphology and cell surface makers as primary MSCs. IMSCs possessed better ability of proliferation and anti-senescence compared with primary MSCs, while maintained multilineage differentiation capacity. Neural-like cells derived from IMSCs had similar expressions of neural-specific genes, protein expression patterns and resting membrane potential (RMP) compared with their counterparts derived from primary MSCs. There was no bump formation in nude mice subcutaneously injected with IMSCs. IMSCs played same role as primary MSCs to improve learning ability and spatial memory of HIBD rats. Conclusions IMSCs not only retain their features of primary MSCs but also possess the ability of high proliferation and anti-senescence. IMSCs can definitely be induced to differentiate into neuronal cells in vitro and take the place of primary MSCs for cell transplantation therapy without tumorigenesis in vivo. The stable cell line is particularly useful and valuable as an alternative to MSCs in neuronal differentiation and neuroregeneration associated studies.

Mesenchymal stem cells (MSCs) can differentiate into neurons in an appropriate cellular environment. Retinoid signaling pathway is required in neural development. However, the effect and mechanism through retinoid signaling regulates neuronal differentiation of MSCs are still poorly understood. Here, we report that all-trans-retinoic acid (ATRA) pre-induction improved neuronal differentiation of rat MSCs. We found that, when MSCs were exposed to different concentrations of ATRA (0.01-100 micromol/L) for 24 h and then cultured with modified neuronal induction medium (MNM), 1 micromol/L ATRA pre-induction significantly improved neuronal differentiation efficiency and neural-cell survival. Compared with MNM alone induced neural-like cells, ATRA/MNM induced cells expressed higher levels of Nestin, neuron specific enolase (NSE), microtubule-associated protein-2 (MAP-2), but lower levels of CD68, glial fibrillary acidic protein (GFAP), and glial cell line-derived neurotrophic factor(GDNF), also exhibited higher resting membrane potential and intracellular calcium concentration, supporting that ATRA pre-induction promotes maturation and function of derived neurons but not neuroglia cells from MSCs. Endogenous retinoid X receptors (RXR) RXRalpha and RXRgamma (and to a lesser extent, RXRbeta) were weakly expressed in MSCs. But the expression of RARalpha and RARgamma was readily detectable, whereas RARbeta was undetectable. However, at 24 h after ATRA treatment, the expression of RARbeta, not RARalpha or RARgamma, increased significantly. We further found the subnuclear redistribution of RARbeta in differentiated neurons, suggesting that RARbeta may function as a major mediator of retinoid signaling during neuronal differentiation from MSCs. ATRA treatment upregulated the expression of Vimentin and Stra13, while it downregulated the expression of Brachyury in MSCs. Thus, our results demonstrate that pre-activation of retinoid signaling by ATRA facilitates neuronal differentiation of MSCs.

This study examined whether pregnancy-related marginal vitamin A deficiency (MVAD) influences postnatal development of retinoic acid receptors (RARs) and N-methyl-D-aspartate (NMDA) receptor subunit 1 (NR1) in hippocampus of rat pups. Sixteen female rats were randomized equally into control and MVAD groups. Dams and pups were fed with either a normal control diet or one deficient in vitamin A. Eight female pups in each group were killed at 1 day, 2 weeks, 4 weeks and 8 weeks after birth, respectively. Serum retinol levels were monitored. The messenger RNA (mRNA) and protein expressions and subcellular localization of RARα, RARβ and NR1 in postnatal hippocampus were detected. At 1 day, 2 weeks and 8 weeks after birth, serum retinol levels in the MVAD group were significantly lower than those in the control group. Results of Morris water maze test at 7 weeks of age showed that spatial learning and memory in the MVAD group were affected. Vitamin A deficiency resulted in decreased mRNA levels of RARα, RARβ and NR1 (P.05). The protein level of RARα and NR1 in the MVAD group was lower than that of the control group (P.05). There was no significant difference in RARβ between the groups (P.05). A mass of RARα and NR1 colocalized in hippocampal cell cytoplasm on postnatal day 1. Our data suggested that vitamin A deficiency in pregnancy may affect the postnatal expression of RARα and NR1, affecting learning and memory function in the hippocampus and synaptic plasticity of the calcium signaling pathway.

ObjectiveTo assess the impact of urbanisation on the prevalence of the metabolic syndrome in Chinese adults.DesignAs part of a community-based cross-sectional survey conducted in 2002, a sample from rural and urban populations in East China was obtained. The metabolic syndrome is defined by the National Cholesterol Education Program Adult Treatment Panel III criteria (ATP III) and the modified ATP III, which recommended a lower waist circumference cut-off for Asians.SettingField sites in Jiangxi and Anhui provinces and the Jing'an District of Shanghai, China.SubjectsA total of 529 non-pregnant, non-lactating urban and rural adults, aged 20–64 years without diagnosed diabetes.ResultsDwelling in urban areas was associated with higher dietary fat intake and slightly lower total energy intake, and with significantly lower occupational physical activity. Using the ATP III criteria, the prevalence of the metabolic syndrome was significantly higher for urban than rural men (12.7 vs. 1.7%, P P = 0.17). These urban–rural differences were greatly enhanced when the modified ATP III criteria for the syndrome were used, for men (34.3 vs. 2.7%, P P = 0.07). The Asian waist circumference cut-offs (90 and 80 cm for men and women, respectively) had a better combination of sensitivity and specificity in identifying other metabolic disorders, which included high glucose, high blood pressure, high triglycerides and low high-density lipoprotein cholesterol, for this population.ConclusionFor the Chinese population, urban dwelling was associated with higher prevalence of the metabolic syndrome, especially in men.

Linear growth retardation (stunting) is prevalent (10-80%) in developing countries. It takes place between 6 and 18 mo of age and can be characterized by the delayed onset of the childhood phase of growth. The objective of this work was to study the existence of stunting in early life and, when present, its lifelong effects on final height in a normal healthy population in a developed country. A total of 2432 children with longitudinal height data from birth to adulthood were analyzed. The series represents a normal healthy population-based sample of full-term children born around 1974 in Goteborg, Sweden. We found the incidence of stunting to be about 10% as indicated by delayed onset of the childhood phase of growth (>12 mo of age): 14% in boys and 8% in girls. In a multiple linear regression model, final height was described by length at 6 mo of age, age at onset of the childhood phase of growth, and age at peak height velocity during puberty. All three measures contributed significantly (p < 0.05) to final height with a total R2 of 0.33. A final height difference of 5.0 cm could be explained by abnormal onset of the childhood phase of growth in this normal population. The timing of the childhood onset of growth did not correlate with the timing of puberty. For the first time, we can conclude that stunting in early life is not only a problem in developing countries. Its importance on final height achieved is clearly shown, even after controlling for the timing of puberty.

INTRODUCTION: Vitamin A is an essential micronutrient for brain development. Marginal vitamin A deficiency (MVAD) remains a subclinical public health problem in children, but little is known about the mechanism by which it affects brain development beginning from embryonic period and early postnatal period. OBJECTIVE: The objective of this study was to study the effects of MVAD on the hippocampal CA1 long-term potentiation (LTP) in young rats. METHODS: The MVAD group was fed a vitamin A–deficient diet (400 IU/kg vitamin A), and the control group was fed a vitamin A–sufficient diet (6500 IU/kg vitamin A) at 3 weeks before coitus. Serum vitamin A was assessed by high-performance liquid chromatography. Hippocampal CA1 LTP was detected by electrophysiologic technique, and the ultrastructure of synapses was observed by electron microscope. RESULTS: The changes of field excitatory postsynaptic potentials slope (25.4% ± 2.01%) in MVAD rats aged 7 weeks was much lower than that in the control group (57.5% ± 8.6%). The changes of slope of field excitatory postsynaptic potentials induced by MVAD in young rats could be replenished after addition of retinoic acid (RA); however, LTP impairment was observed again after addition of RA antagonist into the solution of the control group. No differences of LTP were found after addition of FeSO4 or ZnSO4. The curvature of the synaptic interface of the MVAD group was less than that of the MVAD group that was supplemented with RA and of the control group. CONCLUSIONS: MVAD during the embryonic and early postnatal period can directly impair the hippocampal CA1 LTP of young rats.