ABSTRACT
In the nitrogen-limited subtropical gyres, diazotrophic cyanobacteria, including Crocosphaera, provide an essential ecosystem service by converting dinitrogen (N2) gas into ammonia to support primary production in these oligotrophic regimes. Natural gradients of phosphorus (P) and iron (Fe) availability in the low-latitude oceans constrain the biogeography and activity of diazotrophs with important implications for marine biogeochemical cycling. Much remains unknown regarding Crocosphaera's physiological and molecular responses to multiple nutrient limitations. We cultured C. watsonii under Fe, P, and Fe/P (co)-limiting scenarios to link cellular physiology with diel gene expression and observed unique physiological and transcriptional profiles for each treatment. Counterintuitively, reduced growth and N2 fixation resource use efficiencies (RUEs) for Fe or P under P limitation were alleviated under Fe/P co-limitation. Differential gene expression analyses show that Fe/P co-limited cells employ the same responses as single-nutrient limited cells that reduce cellular nutrient requirements and increase responsiveness to environmental change including smaller cell size, protein turnover (Fe-limited), and upregulation of environmental sense-and-respond systems (P-limited). Combined, these mechanisms enhance growth and RUEs in Fe/P co-limited cells. These findings are important to our understanding of nutrient controls on N2 fixation and the implications for primary productivity and microbial dynamics in a changing ocean.
Subject(s)
Cyanobacteria , Phosphorus , Phosphorus/metabolism , Nitrogen/metabolism , Nitrogen Fixation/physiology , Iron/metabolism , Ecosystem , Seawater/microbiology , Cyanobacteria/metabolismABSTRACT
BACKGROUND: Xiaoyaosan (XYS), a classic traditional Chinese medicine (TCM) prescription that contained eight Chinese herbs, has been used for treating depression for thousands of years. Yet, the underlying mechanisms are still unclear, which need to be investigated from various perspectives. Disassembling a prescription is one of the effective approaches to study the effects and the mechanisms of TCM prescriptions. By disassembling the prescription, we can find effective combinations of individual herbs to simplify the scale of a given prescription. Accordingly, herein, XYS was disassembled into Shugan and Jianpi groups. PURPOSE: This study aimed to explore the anti-depressive effects of XYS and its disassembled groups on the digestive system functions and the cecal microbiota of rats. METHODS: XYS was divided into two efficacy groups, i.e., the Shugan (SG) and the Jianpi (JP) groups. A depression model was applied by using the chronic unpredictable mild stress (CUMS) method. Various classic behavioral tests were performed to assess the anti-depressive effects of the XYS, the SG, and the JP. Afterward, the effects of the three groups on the digestive system functions and the cecum microbiota of depression rats were evaluated. On top of this, correlation analyses between behavioral and digestive system function indexes and cecum microbiota were conducted. RESULTS: The XYS, the SG, and the JP had significant callback effects on depressive behaviors and gastrointestinal dysfunctions of CUMS rats. The compositions of the gut bacterial community were variable among the five groups. The community composition of the SG was the most similar to that of NC, followed by the XYS and the JP. At phylum, family, and genus levels, 31 potential microbial biomarkers associated with CUMS were identified. Twenty biomarkers were significantly reversed by the SG while 16 and 11 biomarkers were reversed by the XYS and the JP, respectively. The results of degrees of regulatory effects showed that the SG had the highest efficacy index (EI) than the XYS and the JP. CONCLUSION: Regarding the regulation of cecal microbiota of depression rats, the SG treatment was better than XYS and JP. Therefore, SG could be used individually for the clinical treatment of depression, especially in patients with gastrointestinal and gut microbiota disorders.
Subject(s)
Antidepressive Agents/pharmacology , Depression/drug therapy , Depression/microbiology , Drugs, Chinese Herbal/pharmacology , Gastrointestinal Microbiome/drug effects , Animals , Behavior, Animal/drug effects , Biomarkers/analysis , Drugs, Chinese Herbal/chemistry , Dysbiosis/drug therapy , Dysbiosis/microbiology , Gastric Emptying/drug effects , Gastrointestinal Microbiome/genetics , Gastrointestinal Microbiome/physiology , Male , Rats, Sprague-Dawley , Stress, Psychological/drug therapy , Stress, Psychological/microbiologyABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: Xiaoyaosan (XYS), a representative and classic prescription in traditional Chinese medicines (TCMs), has been used for thousands of years for treating depression. The anti-depression effect of XYS has been demonstrated both clinically and experimentally. However, it is still unclear that whether XYS could regulate the abnormalities of gut microbiota and metabolites of cecum induced by depression, and in which way. This study aimed to explore the underlying mechanism of the anti-depressant effects of XYS from the perspective of cecal microbiota and metabolites. MATERIALS AND METHODS: Chronic unpredictable mild stress (CUMS)-induced depression-like rats were used as the depression animal model. Various classic behavioral tests were performed to assess the anti-depressant effects of XYS. Additionally, the composition, the richness, and the diversity of the cecum microbiota were assessed by 16S rRNA gene sequencing technology. Besides, the metabolic profiling of cecum samples was analyzed by 1H-NMR metabolomics. Multivariate data analysis was then applied to screen the differential metabolites and to characterize the changes in cecum metabolites. Moreover, a correlation analysis between differential metabolites and crucial microbiota was conducted. RESULTS: XYS significantly improved depressive behaviors and the abnormal diversity of cecum microbiota induced by CUMS. At the phylum level, XYS could significantly increase the abundance of Firmicutes while decrease the abundance of Actinobacteria in depressed rats. XYS significantly regulated the abundances of 9 out of 13 potential microbial biomarkers at the genus level. Cecal metabolomics showed that XYS could also regulate the abnormal levels of alanine, proline, lactate, and valine of depression rats. CONCLUSIONS: This study revealed, for the first time, from the perspectives of microbiota and cecum metabolites, the anti-depression mechanisms of XYS. This study is of significance for not only comprehensively understanding the anti-depression effects and mechanisms of XYS, but also for providing a research approach for revealing the underlying mechanisms of action of TCMs, i.e. to apply a combination of 16S rRNA gene sequencing and metabolomics.
Subject(s)
Antidepressive Agents/pharmacology , Cecum/microbiology , Depression/drug therapy , Drugs, Chinese Herbal/pharmacology , Metabolome/drug effects , Microbiota/drug effects , Amino Acids/metabolism , Animals , Antidepressive Agents/therapeutic use , Behavior, Animal/drug effects , Biomarkers/metabolism , Body Weight/drug effects , Depression/etiology , Disease Models, Animal , Drugs, Chinese Herbal/therapeutic use , Food Preferences/drug effects , Male , Medicine, Chinese Traditional , Microbiota/genetics , Motor Activity/drug effects , Principal Component Analysis , Rats, Sprague-Dawley , Stress, Psychological/complications , Stress, Psychological/microbiologyABSTRACT
Six aerobic Gram-negative bacteria were isolated from seawater in Guangdong Province, P.R. China. Cells were observed to be Gram-negative, aerobic, non-motile and non-spore forming. Growth of the designated type strain 19X3-30T occurred at a temperature range of 14-37 °C (optimum, 28 °C), a pH range of 6.0-8.0 (optimum, pH 7) and up to 7.5% NaCl (optimum, 1.5%; w/v), and was enhanced by CO2 and L-cysteine supplementation. The major polar lipids identified in strain 19X3-30T were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. The principal cellular fatty acids profile showed the presence of anteiso-C15:0, anteiso-C17:0 and C18:0 (> 8% of total fatty acids), and the respiratory quinone was ubiquinone 8 (UQ-8). According to the analysis of 16S rRNA gene sequences, these strains represented a novel species within the family Fastidiosibacteraceae, sharing maximum similarities with Cysteiniphilum litorale DSM 101832T (96.6%) and Cysteiniphilum halobium DSM 103992T (95.3%). Phylogenetic dendrograms based on 16S rRNA gene and protein marker genes from the genomic sequences both indicated that the strains formed a monophyletic lineage closely linked to the genus Cysteiniphilum, which was also supported by the UPGMA dendrogram based on the MALDI-TOF MS profile. The genomic DNA G + C contents of six strains ranged from 38.0% to 38.1%. Based on different taxonomic genomic metrics, phylogeny and phenotypic features, we propose that the strains warrant the assignment to a novel species, for which the name Cysteiniphilum marinum sp. nov. is proposed. The type strain is 19X3-30T (= KCTC 82154T = CGMCC 1.18585T).
Subject(s)
Phospholipids , Seawater , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids , Gammaproteobacteria , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
Compatibility investigations are vital but also the most difficult aspect of traditional Chinese medicine (TCM) prescriptions. Previous researches have demonstrated that Xiaoyaosan (XYS) is an effective treatment for depression. However, the compatibility rules of XYS and the underlying mechanisms remain unclear. Herein, we combined network pharmacology and serum metabolomics to investigate the scientific connotations and the compatibility effects of XYS. First, the Traditional Chinese Medicine Systems Pharmacology (TCMSP) and the GeneCards databases were applied to identify the chemical components and the putative targets of XYS, and its efficacy groups. We then analyzed the herb-component-target pathways and constructed PPI networks. Potential pathways were determined by gene ontology (GO) and pathway enrichment analyses. Additionally, a chronic unpredictable mild stress (CUMS) model was applied on rats. A proton nuclear magnetic resonance spectrometry (1H NMR) based serum metabolomics was then used to identify potential metabolites and the corresponding pathways that were involved in depression and the potential anti-depression effects of XYS and its efficacy groups. A total of 121 components of XYS and 111 targets were associated with depression. Additionally, we screened 105 targets of 24 components in Shugan (SG) group and 106 targets of 95 components in Jianpi (JP) group. 13 targets were common to all three groups. Protein-protein interaction network analysis showed that Caspase-3 and Nitric oxide synthase-3 were nodes that overlapped with proteins known to be associated with depression. Pathway enrichment analysis further indicated that the putative targets of XYS and its efficacy groups mostly participated in pathways associated with cancer. Also, XYS and its efficacy groups significantly reversed abnormalities of serum metabolites and metabolic pathways, but to different degrees. Overall, different efficacy groups of XYS exhibit synergistic anti-depression effects and contribute to the whole prescription against depression. This study shows that the combination of network pharmacology and metabolomics is an effective approach to demonstrate scientific connotations and compatibility of TCMs from a holistic perspective.
Subject(s)
Antidepressive Agents , Drugs, Chinese Herbal , Animals , Depression/drug therapy , Drugs, Chinese Herbal/pharmacology , Metabolomics , RatsABSTRACT
Philadelphia chromosome-positive (Ph+) is considered as a high risk of acute lymphoblastic leukemia (ALL). Tyrosine kinase inhibitors (TKIs) are tailored drug for Ph+ ALL, but Ik6 is associated with TKI resistance and poor outcome of Ph+ ALL. In the present study, we investigated the potential benefit of combination therapy with imatinib and Huaier extract, a traditional Chinese medicine, in Ik6+ Ph+ ALL. The Ik6+ Ph+ -ALL cell lines Sup-B15 or BV173 were treated with Huaier extract, imatinib or the combination of the two. Analysis of cell proliferation showed that the combined treatment of imatinib and Huaier extract exhibited a greater effect on cell inhibition. Using flow cytometry and Western blot, enhanced effects on the induction of cell apoptosis were observed. The combination of the two drugs also exhibited a significant effect in decreasing the protein and enzymatic activity levels of BCR-ABL. The molecular mechanisms may be involved in BCR-ABL related pathways, including the inactivation of p-AKT, p-STAT5, p-mTOR and p-Lyn. Consistent with the in vitro results, the combination of Huaier extract and imatinib inhibit the growth and infiltration of xenografted tumors. Taken together, our findings show that Huaier extract enhances the anticancer efficacy of imatinib in Ik6+ Ph+ ALL Further, it also provides a potential clinical application in the treatment of refractory Ph+ ALL.
Subject(s)
Complex Mixtures/chemistry , Ikaros Transcription Factor/metabolism , Imatinib Mesylate/therapeutic use , Philadelphia Chromosome , Plant Extracts/therapeutic use , Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Animals , Apoptosis/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Disease Models, Animal , Fusion Proteins, bcr-abl/metabolism , Humans , Imatinib Mesylate/pharmacology , Male , Mice, Inbred BALB C , Plant Extracts/pharmacology , Signal Transduction/drug effects , Trametes , Treatment OutcomeABSTRACT
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.
Subject(s)
Apoptosis/drug effects , Hypoxia-Ischemia, Brain/drug therapy , Hypoxia-Ischemia, Brain/pathology , Mitochondria/metabolism , Vitamin A/pharmacology , Vitamin A/therapeutic use , Animals , Caspases/metabolism , Cells, Cultured , Dietary Supplements , Female , Glucose/deficiency , Hippocampus/drug effects , Hippocampus/metabolism , Hippocampus/pathology , Hypoxia-Ischemia, Brain/physiopathology , Learning , Membrane Potential, Mitochondrial/drug effects , Mitochondria/drug effects , Neurons/drug effects , Neurons/metabolism , Neurons/pathology , Oxygen , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats, Sprague-Dawley , Retinoic Acid Receptor alpha/metabolism , Signal Transduction/drug effects , Spatial Memory/drug effects , Tretinoin/pharmacology , Tretinoin/therapeutic use , Vitamin A Deficiency/drug therapy , Vitamin A Deficiency/pathology , bcl-2-Associated X Protein/metabolismABSTRACT
OBJECTIVE: To assess the efficacy and safety of Saw Palmetto Extract Capsules in the treatment of benign prostatic hyperplasia (BPH). METHODS: We conducted a multi-centered open clinical study on 165 BPH patients treated with Saw Palmetto Extract Capsules at a dose of 160 mg qd for 12 weeks. At the baseline and after 6 and 12 weeks of medication, we compared the International Prostate Symptom Scores (IPSS), prostate volume, postvoid residual urine volume, urinary flow rate, quality of life scores (QOL), and adverse events between the two groups of patients. RESULTS: Compared with the baseline, both IPSS and QOL were improved after 6 weeks of medication, and at 12 weeks, significant improvement was found in IPSS, QOL, urinary flow rate, and postvoid residual urine. Mild stomachache occurred in 1 case, which necessitated no treatment. CONCLUSION: Saw Palmetto Extract Capsules were safe and effective for the treatment of BPH.
Subject(s)
Plant Extracts/therapeutic use , Prostatic Hyperplasia/drug therapy , Capsules , Humans , Male , Plant Extracts/adverse effects , Quality of Life , SerenoaABSTRACT
Single-basidiospore isolates (SBIs) were obtained from field isolates of Thanatephorus cucumeris (Rhizoctonia solani) AG-1 IC and AG-2-2 IV. Formation of distinctive tufts, a recognized feature of heterokaryon synthesis, was observed, and isolates derived from hyphal-tipped tuft hyphae were obtained following pairings between various strains. Three distinctive types of tufts were formed: the fibrous type of mating-compatible homokaryon-homokaryon (Hom-Hom) pairings, the sparse type between heterokaryon-homokaryon (Het-Hom) pairings originating from one parent, and the compact type between Het-Hom pairings originating from different parents. Amplified Fragment Length Polymorphism (AFLP) profile of fingerprints of these tuft isolates verified that they were all heterokaryotic. Because of heterokaryotic vigor, the growth and pathogenicity of the majority of tuft isolates increased compared with their contributing SBIs. New somatic compatibility groups (SCGs) that were different from parental field isolates occurred following heterokaryon formation within T. cucumeris. Tuft isolates produced by Hom-Hom and Het-Hom pairings among isolates of different parents yielded no somatic compatibility with the original parent isolates and a high frequency of new SCGs (62-100%). This was in contrast to those produced by Hom-Hom and Het-Hom pairings among isolates with a common parent that yielded only 12-37% new SCGs. The SCG diversity of R. solani in the field may be attributed to new fitter heterokaryons formed between a heterokaryon of one pair of parents and a homokaryon of another parent pair. This mechanism greatly contributes to genetic diversity in the field and accounts for the failure to recover the expected distribution of SCGs from a field population.
Subject(s)
Genetic Variation , Plant Diseases/microbiology , Rhizoctonia/genetics , Beta vulgaris/microbiology , Brassica/microbiology , DNA, Fungal/genetics , Genes, Mating Type, Fungal , Pinus/microbiology , Poaceae/microbiology , Polymorphism, Restriction Fragment Length , Rhizoctonia/growth & development , Rhizoctonia/isolation & purification , Spores, Fungal/genetics , Spores, Fungal/growth & developmentABSTRACT
This study is to clarify the impact of vitamin A or vitamin A combined with other micronutrients supplementation on anemia and growth in preschoolers. In the present study, a total of 290 preschoolers, aged 36-72 months old were randomly assigned to 3 treatment groups: vitamin A (A group), vitamin A plus zinc (AZ group), and vitamin A combined with additional multiple-micronutrient (AMM group). After 6-month supplementation, the height and height-for-age z-score gains of the AZ group were significantly higher than the other groups; the weight gain of the AMM group was greater than the other groups. Compared with baseline values, the concentrations of hemoglobin, and zinc at the end significantly increased in all 3 groups. The incremental concentrations of hemoglobin in the AMM group were significant higher than in the other two groups. Furthermore, the incremental concentrations of serum retinol in the AMM group, and the increase in serum zinc concentrations in the AZ group were significantly higher, respectively, than in the other groups. These 3 kinds of supplements in the present study are effective in enhancing height gains and are effective in reducing the prevalence of anemia. Supplementation of zinc plus vitamin A is a better way for improving children's height and height-for-age z-score. Vitamin A combined with multiple-micronutrient is more effective in improving the hemoglobin concentrations in preschool children.
Subject(s)
Anemia/prevention & control , Body Height/drug effects , Body Weight/drug effects , Micronutrients/therapeutic use , Vitamin A/therapeutic use , Zinc/therapeutic use , Anemia/blood , Anemia/epidemiology , Biomarkers/blood , Child , Child, Preschool , China/epidemiology , Dietary Supplements , Female , Follow-Up Studies , Hemoglobins/drug effects , Humans , Male , Prevalence , Trace Elements/therapeutic use , Treatment Outcome , Vitamin A/blood , Vitamins/therapeutic use , Zinc/bloodSubject(s)
Acupuncture Therapy , Diarrhea, Infantile/therapy , Child, Preschool , Female , Humans , Infant , MaleABSTRACT
ETHNOPHARMACOLOGICAL RELEVANCE: The flowers of Gentiana veitchiorum has been widely used in decoction form in the traditional medicine of Tibet against tussis, tracheitis, angina for their anti-inflammatory, antimicrobial and alexipharmic properties. AIM OF THE STUDY: The aim of current study was to evaluate the therapeutic effects of Feining, a Chinese herbal formula (national invention patent: ZL200510042636.3) against pulmonary injuries and to clarify the mechanisms involved. MATERIALS AND METHODS: Experimental pulmonary injuries were induced by bleomycin (BLM) in rats with or without subsequent treatment of Feining or prednisone as positive control. The pulmonary injuries were evaluated by histological analysis. Also, the levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH) and hydroxyproline (Hyp) in the lung tissue were determined. To clarify one of the possible active principles responsible for Feining, high performance liquid chromatography-diode array detector-mass spectrometry (HPLC-DAD-MS) method was applied to identify the components of Gentiana veitchiorum, one of major ingredients of Feining. RESULTS: Feining significantly improved lung alveolitis scores and reduced the Hyp content of lungs, which is an index of collagen accumulation. Moreover, Feining played a role against the oxidative damages by decreasing the MDA level, whereas increasing SOD and GSH activity, which correlated with oxidation resistance and scavenging of free radicals. In addition, Feining alleviated inflammatory lung injury by decreasing tumor necrosis factor-α (TNF-α) expression. HPLC-DAD-MS analysis revealed that there was 1.97% gentiopicroside in Gentiana veitchiorum. CONCLUSION: Feining has certain therapeutic effects against pulmonary injuries.
Subject(s)
Bleomycin/toxicity , Gentiana/chemistry , Lung Diseases/drug therapy , Plant Extracts/therapeutic use , Animals , Chromatography, High Pressure Liquid , Female , Iridoid Glucosides/analysis , Lung Diseases/chemically induced , Male , Rats , Rats, Sprague-Dawley , Spectrometry, Mass, Electrospray IonizationABSTRACT
OBJECTIVE: Improvement of hemoglobin and serum retinol and facilitation of the mobilization of iron storage were achieved with a multiple-micronutrient-fortified diet in preschoolers for 6 mo in a suburb of Chongqing, China. We investigated whether fortification with multiple micronutrients in a diet for preschool children results in changes in children's infectious morbidity compared with diets fortified solely with vitamin A and with vitamin A plus iron. METHODS: From December 2005 to June 2006, 226 2- to 6-y-old preschool children were recruited from three nurseries 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. The secondary functional outcomes, morbidity of diarrhea and respiratory infection, were collected during supplementation. RESULTS: The groups were comparable concerning compliance and loss to follow-up. There was evidence of a lower incidence rate of respiratory-related illnesses, diarrhea-related illness, fewer symptoms of runny nose, cough, and fever, and shorter duration of respiratory-related illnesses and cough for children in group III compared with children in groups I and II. However, there was no significantly or clinically important difference between children in groups I and II. CONCLUSION: The beneficial effects on infectious morbidity over 6 mo, in addition to some biochemical improvements, highlight the potential of this micronutrient-fortified seasoning powder supplied in a diet for preschool children.
Subject(s)
Diarrhea/prevention & control , Dietary Supplements , Food, Fortified , Iron, Dietary/therapeutic use , Micronutrients/therapeutic use , Respiratory Tract Infections/prevention & control , Vitamin A/therapeutic use , Child , Child, Preschool , China/epidemiology , Cough , Diarrhea/epidemiology , Drug Therapy, Combination , Female , Fever , Humans , Incidence , Male , Respiratory Tract Infections/complications , Respiratory Tract Infections/epidemiology , Rhinitis , SpicesABSTRACT
OBJECTIVE: To improve the efficiency of primary culture of hippocampal neurons and obtain highly purified neurons with good in vitro growth and minimal risk of contamination. METHODS: The hippocampal neurons of neonatal Wistar rats were isolated and the single cell suspension was prepared by mechanical trituration and sedimentation in stead of trypsin digestion and filteration. Twenty-four hours after the cell plating, the culture medium was removed and replaced by serum-free DMEM/F12 with B27 supplementation. Half of the culture medium was changed 2-3 times every week. The morphological changes of the neurons were observed under inverted phase-contrast microscope. Immunofluorescence staining for NSE was performed to identify the neurons, and the purity of neurons was calculated. The hippocampal neurons were stained with calcium-sensitive fluorescent dye to monitor the effect of KCl on neuronal excitability by a calcium imaging system. RESULTS AND CONCLUSION: This simplified method is time-saving and cost-effective for primary culture of hippocampal neurons with reduced risk of contamination, and the neurons obtained showed high uniformity, purity and long-term viability.
Subject(s)
Hippocampus/cytology , Neurons/cytology , Neurons/physiology , Primary Cell Culture/methods , Action Potentials , Animals , Animals, Newborn , Culture Media, Serum-Free , Female , Male , Rats , Rats, WistarABSTRACT
OBJECTIVE: To 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. DESIGN: A randomised double-masked population-based field interventional trial with a positive control group. SETTING: Banan district of Chongqing, China. SUBJECTS: A 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. RESULTS: All 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. CONCLUSIONS: Data 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.
Subject(s)
Anemia/diet therapy , Child Development/drug effects , Food, Fortified , Hemoglobins/analysis , Vitamin A Deficiency/diet therapy , Vitamin A/administration & dosage , Anemia/blood , Anemia/epidemiology , Body Height/drug effects , Body Height/physiology , Body Weight/drug effects , Body Weight/physiology , Bread , Child , Child Development/physiology , Child, Preschool , China/epidemiology , Double-Blind Method , Female , Humans , Male , Nutrition Policy , Nutritional Status , Vitamin A/therapeutic use , Vitamin A Deficiency/blood , Vitamin A Deficiency/epidemiologyABSTRACT
BACKGROUND: 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. METHODS: 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. RESULTS: 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<0.01). Children with VAD had a significantly increased risk for anemia (odds ratio, 2.56; 95% confident interval, 1.15-5.70). In all 108 children with anemia, only 42 were related to VAD and 12 related to iron deficiency, suggesting that almost half of the anemia children cannot be explained solely by iron deficiency or VAD. CONCLUSIONS: Vitamin A and iron deficiency are still public health problems in some localities of China. Public health interventions in anemia control should be used to eliminate deficiencies of vitamin A, iron, and other micronutrients by deliberate supplementation. Attention must be paid to such deficiencies in high-risk groups, especially in preschool children.
Subject(s)
Anemia, Iron-Deficiency/epidemiology , Child , Child Development , Child, Preschool , China , Cross-Sectional Studies , Female , Humans , Male , Nutritional Status , Suburban Population/statistics & numerical data , Vitamin A Deficiency/epidemiologyABSTRACT
UNLABELLED: 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.
Subject(s)
Lung/embryology , Lung/growth & development , Pregnancy Complications , Vitamin A Deficiency/complications , Animals , Animals, Newborn/growth & development , Body Weight , Collagen/analysis , Diet , Elastin/analysis , Female , Fetal Nutrition Disorders , Lung/chemistry , Organ Size , Pregnancy , Rats , Rats, Wistar , Vitamin A/administration & dosage , Vitamin A/blood , Vitamin A Deficiency/drug therapyABSTRACT
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 (p<0.05) and retinol binding protein from 17.0 (12.6, 25.6) mg/L to 31.6 (24.4, 44.0) mg/L (p<0.05) and to mobilize the stored iron in the liver (p<0.05). 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 (p>0.05). 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.
Subject(s)
Anemia, Iron-Deficiency/diet therapy , Growth/drug effects , Iron, Dietary/therapeutic use , Micronutrients/therapeutic use , Vitamin A Deficiency/diet therapy , Vitamin A/therapeutic use , Vitamins/therapeutic use , Body Size , Child, Preschool , China , Drug Therapy, Combination , Female , Ferritins/blood , Food, Fortified , Hemoglobins/metabolism , Humans , Iron, Dietary/pharmacology , Male , Micronutrients/pharmacology , Retinol-Binding Proteins/metabolism , Vitamin A/metabolism , Vitamins/pharmacologyABSTRACT
OBJECTIVE: In recent years, some experiments on vitamin A-deprived animals reveal a progressive and ultimately profound impairment of hippocampal CA1 area's long-term potentiation and these losses are fully reversible by dietary vitamin A replenishment in vivo. Our previous study revealed that marginal vitamin A deficiency (MVAD) beginning from embryonic period impairs learning, memory and long-term potentiation (LTP) in young rats. But the losses might not be reversible if the vitamin A supplementation is late, especially when the critical period of hippocampus development is missed. The present study aimed to observe the recovery of learning and memory in vitamin A marginally deficient young rats after early intervention with vitamin A supplementation and begin to study the mechanism. METHODS: Rats were divided into control, MVAD, vitamin A intervention 1 (VAI1) and VAI2 groups in this study. In control group (10 young rats) the dams and pups were fed with normal diet (VA 6500 U/kg). In MVAD group (19 young rats) the dams and pups were fed with MVAD diet (VA 400 U/kg). In VAI1 group (10 young rats) the dams were fed with MVAD diet till day 14 of pregnancy, then were fed with normal diet and the pups were fed with normal diet. In VAI2 group (13 young rats) the dams were fed with MVAD diet till delivery, then were fed with normal diet and the pups were fed with normal diet too. All the young rats were killed at the age of 7 weeks. During the last week of the experiment, the shuttle box active avoidance reaction tests were carried out. At week 7, the hippocampal CA1 LTP was detected by electrophysiological technique. The expression of RAR-alpha, RAR-beta, RXR-beta, RXR-gamma, RC3 and tTG mRNA was detected by using semi-quantified RT-PCR in hippocampus. RESULTS: (1) The times to reach the learning standard in MVAD group (45.6 +/- 12.1) were more than those in control group (17.1 +/- 4.4) (P < 0.01), in both VAI1 group (20.8 +/- 3.1) and VAI2 group (22.1 +/- 4.0) were more than those in group MVAD (P < 0.01), and there were no significant differences among groups VAI1, VAI2 and control (P > 0.05) in active avoidance reaction tests. (2) The changes of field excitatory postsynaptic potentials (fEPSP) slope for MVAD group [(22.9 +/- 9.4)%] and VAI2 group [(39.1 +/- 4.33)%] were less than that of control group [(57.5 +/- 27.3)%], respectively (P < 0.05). No significant difference was found between VAI1 and control group (P > 0.05). (3) The expression of RAR-beta and RXR-beta mRNA decreased by 48.72% and 37.84% respectively (P < 0.05) compared with control, but the expression of RAR-beta mRNA in group VAI1 was higher than that in group MVAD (P = 0.065). The expression of RC3 mRNA in MVAD group was lower than that in control (P = 0.061) and RAR-alpha mRNA in MVAD group was higher than that in control (P = 0.061). The expression of RXR-gamma and tTG mRNA had no significant difference among different groups as determined with semi-quantified RT-PCR in hippocampus. CONCLUSION: Early vitamin A intervention may make the impaired learning and memory behavior due to marginal vitamin A deficiency recover to the normal level in young rats, but lip losses in group VAI2 might not be reversible. Vitamin A may modulate the expression of RC3 mRNA by affecting RAR-alpha, RAR-beta and RXR-beta to influence the LTP, learning and memory.
Subject(s)
Animal Nutritional Physiological Phenomena , CA1 Region, Hippocampal/metabolism , Learning/drug effects , Memory/drug effects , Rats/growth & development , Vitamin A Deficiency/drug therapy , Vitamin A/therapeutic use , Animals , Long-Term Potentiation/drug effects , Neurogranin/genetics , Neurogranin/metabolism , RNA, Messenger/metabolism , Receptors, Retinoic Acid/genetics , Receptors, Retinoic Acid/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Transglutaminases/genetics , Transglutaminases/metabolismABSTRACT
OBJECTIVE: Previous studies have demonstrated that vitamin A and its active derivatives function as essential competence factors for long-term synaptic plasticity within the adult brain. But little is known if marginal vitamin A deficiency (MVAD) beginning from embryonic period affects the brain development and the ability of learning and memory in young rats. The aim of this study was to identify the effects of MVAD and vitamin A intervention (VAI) on learning, memory and the hippocampal CA1 long-term potentiation (LTP) in young rats. METHODS: Rats were divided into control, MVAD and VAI groups in this study. In control group (10 young rats) the dams and pups were fed with normal diet (VA 6500 IU/kg). In MVAD group (19 young rats) the dams and pups were fed with MVAD diet (VA 400 IU/kg). In VAI group (9 young rats) the dams were fed with MVAD diet and the pups were fed with normal diet from postnatal week 4. All the young rats were killed at the age of 7 weeks. During the last week of the experiment, the shuttle box active avoidance reaction tests were carried out. At week 7, the hippocampal CA1 LTP was detected by electrophysiological technique and relative intensity of fluorescence in cells in hippocampal slices was measured by confocal laser scanning microscopy labeled by fluo-3. RESULTS: (1) The times to reach the learning standard in both VAI group (28.8 +/- 4.1) and MVAD group (45.6 +/- 12.1) were more than control group (17.1 +/- 4.4) (P < 0.01), and that of MVAD group was more than VAI group (P < 0.05) in active avoidance reaction tests. (2) The changes of field excitatory postsynaptic potentials (fEPSP) slope for MVAD group (22.9% +/- 9.4%) and VAI group (29.5% +/- 13.7%) were less than that of control group (57.5% +/- 27.3%), respectively (P < 0.01). No significant difference was found between VAI and MVAD groups (P > 0.05). (3) No significant differences of relative intensity of fluorescence in cells were found among the three groups before the tetanus stimulation. However, the significantly low relative intensity of fluorescence in cells was seen in MVAD (65.1 +/- 17.0) and VAI (85.8 +/- 17.1) groups compared with control group (113.6 +/- 20.5) after the tetanus stimulation (P < 0.01), and that of VAI group was higher than that of MVAD group (P < 0.05). CONCLUSION: MVAD beginning from embryonic period impairs learning, memory and LTP in young rats. But the losses might not be reversible if the vitamin A supplementation is late especially missing the critical period of hippocampus development. According to the experimental data, it is speculated that vitamin A may modulate the influx of calcium ion to influence the LTP and lead to the change of learning and memory.