Maternal dietary deficiencies in folic acid or choline worsen stroke outcomes in adult male and female mouse offspring

Maternal one-carbon metabolism plays an important role in early life programming.There is a well-established connection between the fetal environment and the health status of the offspring.Howeve r,there is a knowledge gap on how maternal nutrition impacts stro ke outcomes in offspring.The aim of our study was to investigate the role of maternal dietary deficiencies in folic acid or choline on stroke outcomes in 3-month-old offspring.Adult female mice were fed a folic acid-deficient diet,choline-deficient diet,or control diet 4 weeks before pregnancy.They we re continued on diets during pregnancy and la ctation.Male and female offspring were weaned onto a control diet and at 2 months of age were subjected to ischemic stroke within the sensorimotor cortex via photothrombotic damage.Mothers maintained on either a folic acid-deficient diet or choline-deficient diet had reduced levels of S-adenosylm ethionine in the liver and S-adenosylhomocysteine in the plasma.After ischemic stro ke,motor function was impaired in 3-month-old offspring from mothers receiving either a folic acid-deficient diet or choline-deficient diet compared to the animals receiving a control diet.In brain tissue,there was no difference in ischemic damage volume.When protein levels were assessed in ischemic brain tissue,there were lower levels of active caspase-3 and hypoxia-inducible factor 1α in males compared to females and betaine levels were reduced in offspring from the mothers receiving a choline-deficient diet.Our results demonstrate that a deficient maternal diet at critical time points in neurodevelopment results in worse stro ke outcomes.This study emphasizes the importance of maternal diet and the impact it can have on offspring health.

Enhancing effect of choline chloride-based deep eutectic solvents with polyols on the aqueous solubility of curcumin–insight from experiment and theoretical calculation

The development of green solvents for enhancing aqueous solubility of drug curcumin remains a challenge. This study explores the enhancing effect of deep eutectic solvents(DESs) on the aqueous solubility of curcumin(CUR) via experiment and theoretical calculation. Choline chloride-based DESs with polyols 1,2-propanediol(1,2-PDO), 1,3-propanediol, ethylene glycol, and glycerol as hydrogen bond donors were prepared and used as co-solvents. The CUR aqueous solubility increased with increasing the DESs content at temperature of 303.15-318.15 K, especially in aqueous ChCl/1,2-PDO(mole ratio 1:4) solutions. The positive apparent molar volume values and reduced density gradient analysis confirmed the existence of strong interactions between CUR and solvent. The van der Waals interactions and hydrogen bonding coexisted in DESs monomer retained the stability of DESs structure after introducing CUR. Moreover,the lower interaction energy of DESs…CUR system than that of the counterpart DESs further proved the strong interaction between CUR and DESs. The lowest interaction energy of ChCl/1,2-PDO…CUR system indicated that this system was the most stable and ChCl/1,2-PDO was promising for CUR dissolution.This work provides efficient solvents for utilizing curcumin, contributing to a deep insight into the interactions between DES and CUR at the molecular level, and the role of DESs on enhancing drugs solubility.

Choline dehydrogenase interacts with SQSTM1 to activate mitophagy and promote coelomocyte survival in Apostichopus japonicus following Vibrio splendidus infection

Previous studies have shown that Vibrio splendidus infection causes mitochondrial damage in Apostichopus japonicus coelomocytes,leading to the production of excessive reactive oxygen species(ROS)and irreversible apoptotic cell death.Emerging evidence suggests that mitochondrial autophagy(mitophagy)is the most effective method for eliminating damaged mitochondria and ROS,with choline dehydrogenase(CHDH)identified as a novel mitophagy receptor that can recognize non-ubiquitin damage signals and microtubule-associated protein 1 light chain 3(LC3)in vertebrates.However,the functional role of CHDH in invertebrates is largely unknown.In this study,we observed a significant increase in the mRNA and protein expression levels of A.japonicus CHDH(AjCHDH)in response to V.splendidus infection and lipopolysaccharide(LPS)challenge,consistent with changes in mitophagy under the same conditions.Notably,AjCHDH was localized to the mitochondria rather than the cytosol following V.splendidus infection.Moreover,AjCHDH knockdown using si RNA transfection significantly reduced mitophagy levels,as observed through transmission electron microscopy and confocal microscopy.Further investigation into the molecular mechanisms underlying CHDH-regulated mitophagy showed that AjCHDH lacked an LC3-interacting region(LIR)for direct binding to LC3 but possessed a FB1 structural domain that binds to SQSTM1.The interaction between AjCHDH and SQSTM1 was further confirmed by immunoprecipitation analysis.Furthermore,laser confocal microscopy indicated that SQSTM1 and LC3 were recruited by AjCHDH in coelomocytes and HEK293T cells.In contrast,AjCHDH interference hindered SQSTM1 and LC3 recruitment to the mitochondria,a critical step in damaged mitochondrial degradation.Thus,AjCHDH interference led to a significant increase in both mitochondrial and intracellular ROS,followed by increased apoptosis and decreased coelomocyte survival.Collectively,these findings indicate that AjCHDH-mediated mitophagy plays a crucial role in coelomocyte survival in A.japonicus following V.splendidus infection.

Assessment of the Effects of Supplemental Rumen Protected B Vitamins and Choline for Periparturient Cows: A Meta-Analysis of 28 Feeding Studies

There are currently no prescribed requirements for B vitamins or choline for dairy cows during the transition period, but many recent studies have shown a variety of benefits from supplying these nutrients to periparturient cows. The purpose of this research effort was to determine the potential benefit of including a rumen protected blend composed of B vitamins (riboflavin, folic acid, vitamin B12) and choline (RPBlend, Jefo Nutrition Inc., St. Hyacinthe, QC, Canada) for dairy cows during the transition period, based on results from 28 on-farm feeding studies (USA-12, Mexico-7, Canada-4, Chile-3, Australia-1, Brazil-1) conducted between 2011 and 2018. All farms participated in monthly herd management record keeping systems and were selected to participate in studies due to their excellent management. Meta-analyses in which risk differences were determined were used to assess the effects of the supplemental RPBlend on health parameters and reproduction. The effect size was used as the determinant of the possible contribution of RPblend on the yields of milk, energy corrected milk (ECM), fat and protein yields during the first four weeks of lactation. Results showed that the inclusion of RPblend reduced (P < 0.05) involuntary culling and mastitis by cows during the first 30 days after calving. There was a tendency (P < 0.10) for reduced retained placenta and metritis. The meta-analyses revealed that the risk of the displaced abomasum and milk fever did not diminish (P > 0.10) with the inclusion of the BPBlend. The incidence rate of subclinical ketosis, determined as blood beta hydroxy butyric acid greater than 1.2 mM was lower (P (than those not supplemented. The proportion of cows confirmed pregnant by 100 days in milk was greater for cows given the BPBlend (P < 0.05). Milk yield and ECM were greater for cows receiving the RPBlend (1.13 and 0.93 kg/cow respectively, P < 0.05). There was no change in fat yield (P > 0.10) while the yield of milk protein was greater (P < 0.05) when the cows received the blend. These results suggest that the inclusion of rumen protected B vitamins and choline can assist cow health, reproduction and production at the start of lactation.

Effect of bilobalide B on cholinergic hippocampal neurons exposed to cholesterol and apolipoprotein E4

BACKGROUND： Extracts of ginkgo biloba leaves have been reported to improve nerve function and activity in Alzheimer＇s disease, which is associated with reduced secretion of cholinergic neurotransmitter in hippocampal neurons. OBJECTIVE： To validate the protective effect of bilobalide B against in vitro injury of cholinergic neurons of the hippocampus induced by combined cholesterol and apoE4 DESIGN, TIME AND SETTING： This randomized, controlled animal experiment was performed in the Pathology Laboratory, Tianjin University of Traditional Chinese Medicine from July 2003 to July 2006. MATERIALS： Neonatal Wistar rats, 1-day-old, both male and female, and mean body mass of 5 g were selected for this study. Cholesterol and apolipoprotein E4 （apoE4） were purchased from Sigma Company （USA）, bilobalide B was purchased from Tianjin Zhongyi Pharmaceutical Factory, batch number 20050312. METHODS： Hippocampal neurons were divided into three groups： a normal control group （routinely added media）, a model group （exposed to media containing 40 mg/L cholesterol and 30 mg/L apoE4 for 24 hours） and a bilobalide B group （exposed to media containing 160 mg/L bilobalide B for 16 hours, and then with addition of 40 mg/L cholesterol and 30 mg/L apoE4 for an additional 24 hours）. MAIN OUTCOME MEASURES： Levels of acetylcholine （ACh） and activity of acetylcholinesterase （ACHE） and choline acetyltransferase （CHAT） in hippocampal neurons were determined by microdosage hydroxylamine colorimetry, hydroxylamine colorimetry and radiological chemistry, respectively. RESULTS： The ACh level was significantly lower in the model group than that in the normal control group （P 〈 0.01）, while it was markedly higher in the bilobalide B group than in the model group （P 〈 0.05）. Activity of AChE was significantly decreased in the model group compared with the normal control group （P 〈 0.05）. However, there was no significant difference between the model group and the bilobalide B group （P 〉 0.05）. Activity of ChAT was significantly lower in the model group than in the normal control group （P 〈 0.01）, while the activity was significantly higher in the bilobalide B group than in the model group （P 〈 0.05）. CONCLUSION： Bilobalide B can enhance the ACh level of hippocampal neurons damaged by combined cholesterol and apoE4, by promoting the synthesis, but not the degradation, of ACh.

Cloning of cDNA Encoding Choline Monooxygenase from Suaeda liaotungensis and Salt Tolerance of Transgenic Tobacco

Betaine is a very effective osmoprotectant found in many organisms. In high plants, betaine is synthesized by oxidation of choline in two sequential steps: choline-->betaine aldehyde-->betaine. The first step is catalyzed by choline monooxygenase (CMO). In this study, the full-length CMO cDNA (1 820 bp) was cloned from halophyte Suaeda liaotungensis Kitag by RT-PCR and RACE. It included a 123 bp 5' UTR, a 368 bp 3' UTR and a 1 329 bp open reading frame encoding a 442-amino-acid polypeptide with 77%, 72% and 74% sequence identity compared to CMOs from spinach, sugar beet and Atriplex hortensis, respectively. The CMO open reading frame (ORF) was cloned and the plant expression vector pBI121-CMO was constructed. It was transferred into tobacco ( Nicotiana tabacum L. ev. 89) via Agrobacterium mediation. PCR and Southern blotting analysis showed that the CMO gene was integrated into tobacco genome. Transgenic tobacco plants contained higher amount of betaine than that of control plants and were able to survive on MS medium containing 250 mmol/L NaCl. Relative electronic conductivity demonstrated less membrane damage in transgenic plants as in the wild type.

Impairment of cognitive function and reduced hippocampal cholinergic activity in a rat model of chronic intermittent hypoxia

The present study established a rat model of chronic intermittent hypoxia （CIH） to simulate obstructive sleep apnea syndrome. CIH rats were evaluated for cognitive function using the Morris water maze, and neuronal pathology in the hippocampus was observed using hematoxylin-eosin staining. In addition, hippocampal choline acetyl transferase （CHAT） and nicotinic acetylcholine receptor （nAChR） expression was determined by immunohistochemistry. Our results revealed necrotic hippocampal neurons, decreased ChAT and nAChR expression, as well as cognitive impairment in CIH rats. These results suggest that hippocampal neuronal necrosis and decreased cholinerqic activity may be involved in CIH-induced cognitive impairment in rats.

Choline Promotes Growth and Tabtoxin Production in a <i>Pseudomonas syringae</i>Strain

Some Pseudomonas syringae pathovars secrete tabtoxin, a monocyclic β-lactam antibiotic, responsible for chlorosis, the principal halo blight symptom in susceptible plants as oats, rye, barley, wheat and sorghum, among other. Here, we demonstrated that the production of tabtoxin in a P. syringae strain increased at least 150%, when choline, betaine or dimethylglycine were used as nitrogen source, or when choline was added as osmoprotectant in hyperosmolar culture media. Besides, we investigated the induction of phosphorylcholine phosphatase (PchP) activity when choline or its metabolites were used as nitrogen sources. PchP is an enzyme involved in Pseudomonas aeruginosa pathogenesis through its contribution to the breakdown of choline-containing compounds of the host cells. Considering these results and that the success of a pathogenic microorganism depends on its ability to survive and proliferate in its target tissue, we propose that choline is one of the plant signals that contribute to establishment of the infection by tabtoxin-producing strains of P. syringae.

Change of choline compounds in sodium selenite-induced apoptosis of rats used as quantitative analysis by in vitro 9.4T MR spectroscopy

AIM: To study liver cell apoptosis caused by the toxicity of selenium and observe the alteration of choline compounds using in vitro 9.4T high resolution magnetic resonance spectroscopy. METHODS: Twenty male Wistar rats were randomly divided into two groups. The rats in the treatment group were intraperitoneally injected with sodium selenite and the control group with distilled water. All rats were sacrifi ced and the livers were dissected. 1H-MRS data were collected using in vitro 9.4T high resolution magnetic resonance spectrometer. Spectra were processed using XWINNMR and MestRe-c 4.3. HE and TUNEL staining was employed to detect and confi rm the change of liver cells. RESULTS: Good 1H-MR spectra of perchloric acid extract from liver tissue of rats were obtained. The conventional metabolites were detected and assigned. Concentrations of different ingredient choline compounds in treatment group vs control group were as follows: total choline compounds,5.08 ± 0.97 mmol/L vs 3.81 ± 1.16 mmol/L (P = 0.05); and free choline,1.07 ± 0.23 mmol/L vs 0.65 ± 0.20 mmol/L (P = 0.00). However,there was no statistical signif icance between the two groups. The hepatic sinus and cellular structure of hepatic cells in treatmentgroup were abnormal. Apoptosis of hepatic cells was confi rmed by TUNEL assay. CONCLUSION: High dose selenium compounds can cause the rat liver lesion and induce cell apoptosis in vivo. High resolution 1H-MRS in vitro can detect diversified metabolism. The changing trend for different ingredient of choline compounds is not completely the same at early period of apoptosis.

Isolation of a choline monooxygenase cDNA clone from Amaranthus tricolor and its expressions under stress conditions

Plants synthesize the osmoprotectant glycine betaine (GB) via choline→betaine aldehyde→glycine be- taine[1]. Two enzymes are involved in the pathway choline monooxygenase (CMO) and betaine aldehyde dehydrogenase (BADH). A full length CMO cDNA (1,643bp) was cloned from Amaranthus tricolor. The open reading frame encoded a 442-amino acid polypeptide, which showed 69% identity with CMOs in Spina- cia oleracea L. and Beta vulgaris L. DNA gel blot analysis indicated the presence of one copy of CMO gene in the A. tricolor genome. The expressions of CMO and BADH proteins in A.tricolor leaves significantly increased under salinization, drought and heat stress (42℃), as determined by immunoblot analysis, but did not respond to cold stress (4℃), or exogenous ABA application. The increase of GB content in leaves was parallel to CMO and BADH contents.

Tolerance of nestin^+ cholinergic neurons in the basal forebrain against colchicine-induced cytotoxicity

In the present study we injected colchicine into the lateral ventricle of Sprague-Dawley rats to investigate the effects of colchicine on the number of different-type neurons in the basal forebrain and to search for neurons resistant to injury. After colchicine injection, the number of nestin^＋ cholinergic neurons was decreased at 1 day, but increased at 3 days and peaked at 14-28 days. The quantity of nestincholinergic neurons, parvalbumin-positive neurons and choline acetyl transferase-positive neurons decreased gradually. Our results indicate that nestin^＋ cholinergic neurons possess better tolerance to colchicine-induced neurotoxicity.

Quantification of choline concentration following liver cell apoptosis using ~1H magnetic resonance spectroscopy

AIM: To evaluate the feasibility of quantifying liver choline concentrations in both normal and apoptotic rabbit livers in vivo, using 1H magnetic resonance spectroscopy (1H-MRS). METHODS: 1H-MRS was performed in 18 rabbits using a 1.5T GE MR system with an eight-channel head/neck receiving coil. Fifteen rabbits were injected with sodium selenite at a dose of 10 μmol/kg to induce the liver cell apoptosis. Point-resolved spectroscopy sequencelocalized spectra were obtained from 10 livers once before and once 24 h after sodium selenite injection in vivo. T1 and T2 relaxation time of water and choline was measured separately in the livers of three healthy rabbits and three selenite-treated rabbits. Hematoxylin and eosin and dUTP-biotin nick end labeling (TUNEL) staining was used to detect and confirm apoptosis. Choline peak areas were measured relative to unsuppressed water using LCModel. Relaxation attenuation was corrected using the average of T1 and T2 relaxation time. The choline concentration was quantified using a formula, which was tested by a phantom with a known concentration. RESULTS: Apoptosis of hepatic cells was confirmed by TUNEL assay. In phantom experiment, the choline concentration (3.01 mmol/L), measured by 1H-MRS, was in good agreement with the actual concentration (3 mmol/L). The average T1 and T2 relaxation time of choline was 612 ± 15 ms and 74 ± 4 ms in the control group and 670 ± 27 ms and 78 ± 5 ms in apoptotic livers in vivo, respectively. Choline was quantified in 10 rabbits, once before and once after the injection with sodium selenite. The choline concentration decreased from 14.5 ± 7.57 mmol/L before sodium selenite injection to 10.8 ± 6.58 mmol/L (mean ± SD, n = 10) after treatment (Z = -2.395, P < 0.05, two-sample paired Wilcoxon test). CONCLUSION: 1H-MRS can be used to quantify liver choline in vivo using unsuppressed water as an internal reference. Decreased liver choline concentrations are found in sodium selenite-treated rabbits undergoing liver cell apoptosis.

3.0 T proton magnetic resonance spectroscopy of the liver: Quantification of choline

AIM: To investigate the normal hepatic magnetic resonance spectroscopy findings choline/lipid2 (Cho/Lip2) associated with age and body mass index (BMI).METHODS: A total of 58 single-voxel proton spectra of the liver were acquired at 3.0 T using the eightchannel phased array abdominal coil as the receiver coil. Consecutive stacks of breath-hold spectra were acquired using the point resolved spectroscopy technique at a short echo time of 30 ms and a repetition time of 1500 ms. The spectra were processed with the SAGE software package. Areas and heights for metabolite resonance were obtained. Student's t test for unpaired data was used for comparisons of shimming, Cho/Lip2, and lipid content. RESULTS: There were significant negative correlations between the Cho/Lip2 peak height ratios and BMI (r=-0.615) and age (r=-0.398) (all P<0.01). Compared with the high-BMI group, the low-BMI group was younger (39.1±13.0 years vs 47.6±8.5 years, t=-2.954,P=0.005); had better water suppression (93.4%±1.4% vs 85.6%±11.6%, t=2.741, P=0.014); had higher Cho/Lip2 peak heights ratio (0.2±0.14 vs 0.05±0.04,t=6.033,P<0.000); and had lower lipid content (0.03±0.08 vs 0.29±0.31, t=-3.309, P=0.004). Compared with the older group, the younger group had better shimming effects (17.1±3.6 Hz vs 22.0±6.8 Hz, t=-2.919, P=0.008); higher Cho/Lip2 peak heights ratios (0.03±0.05vs 0.09±0.12,t=2.4, P=0.020); and lower lipid content (0.05±0.11 vs 0.23±0.32,t=-2.337,P=0.031). Compared with the lowcholine peak group, the high-choline peak group had lower lipid content (0.005±0.002 vs 0.13±0.23, t=-3.796,P<0.000); lower BMI (19.6±2.4vs 23.9±3.0, t=-4.410, P<0.000); and younger age (34.7±10.0 years vs 43.2±12.5 years, t=-2.088, P=0.041). CONCLUSION: Lipid accumulation could result from the increased fat in the body depending on age and BMI. Lipid can mask the resonance signal of choline.

Anionic structural effect in liquid–liquid separation of phenol from model oil by choline carboxylate ionic liquids

Phenolic compounds exist in crude oil as pollutants, and their removal is vital important for the refining and further application of oils. In traditional separation approaches, strong acid and strong base have to be used to remove these compounds, which may cause serious environmental problems. In this work, 19 kinds of cholinium ionic liquids have been developed to separate phenol from model oil by liquid–liquid extraction. Structural effect of anions of the ionic liquids in the separation is systematically investigated. It is found that depending on the chemical structure of ionic liquids, phenol can be removed from toluene with single-step removal efficiency from 86 to 99% under optimal conditions. The type of substituent groups and the-CH_2 number between two carboxylates have obvious effect on the removal efficiency, and more hydrophilic ionic liquids have a stronger extraction performance for phenol. Furthermore, thermodynamic,^(13) C NMR,~1 H NMR and density functional theory calculations have been performed to characterize the extraction process and to understand the extraction mechanism. It is shown that the extraction of phenol from oil to ionic liquid is a favorable process, and this process is mainly driven by enthalpy change. The formation of the hydrogen bond between anion of the ionic liquid and-OH of phenol is the main driving force for the extraction of phenol from oil to the ionic liquids.

Electrochemical Behavior of Niobium Electrodeposited 316 Stainless Steel Bipolar Plate for PEMFC in Choline Chloride Based Ionic Liquids

Niobium was electrodeposited on 316 stainless steel bipolar plates of a fuel cell in water and air-stable choline chloride based ionic liquids. The electrochemical corruption property of bipolar plates in simulated PEMFC environment was investigated. It was showed that the plating film was distributed on the surface of 316 stainless steel like isolated islands with height less than 50 nm. The XPS, XRD results showed that a smooth and strong chemical inert film of Nb O and Nb2O5 was formed on the surface of 316 stainless steel. In simulated cathodic condition, the corrosion potential of Nb coated stainless steel was improved by 244 m V, whilst in an anodic condition, it was improved by 105 m V. The current densities for the coated 316 stainless steel were decreased to 2.479 4 μA·cm-2 from 14.810 μA·cm-2 at-0.1 V and to 0.576 μA·cm-2 from 13.417 μA/·cm-2 at 0.6 V, respectively. It was implied that the niobium coating effectively decreased the corrosion rate. The results of the electrochemical tests indicated that the corrosion resistance of stainless steel was greatly improved after coated with niobium.

Synergistic effects of ginseng stem and leaf-extracted ginsenoside and choline on improving learning and memory in rats Association verification experiment in animals with multiple learning and memory disorders

BACKGROUND： Ginsenoside extracted from the stem and leaf of ginseng （GSL） and choline have both been shown to improve learning and memory functions; however, further studies are needed to understand the synergistic effects of a combination of both. OBJECTIVE： To verify the combined improved synergistic effects of GSL and choline on learning and memory disorders in rats. DESIGN： Control observation. SETTING： Taishan Medical College. MATERIALS： A total of 150 male Kunming mice weighing （204-2） g and 40 healthy male Wistar rats weighing （2204-20） g were provided by the Experimental Animal Department of Jilin University. Animal experimentation received confirmed consent from the local ethic committee. GSL was provided by the Department of Chemistry, Norman Bethune Medical University, and choline was provided by the Third Experiment Factory, Shanghai. METHODS： This study was performed at the Life Science Institute, Taishan Medical College from October 2006 to February 2007. ① Scopolamine-induced learning and memory disorders in rats： Forty rats were randomly divided into control group, model group, combination group （400 mg/kg GSL ＋ 200 mg/kg choline）, GSL （400 mg/kg） group, and choline （200 mg/kg） group, 8 rats/group. Rats were perfused and administrated in the morning, once a day for 14 successive days. Rats in the control group and model group were perfused with 20 mL/kg distilled water and underwent Morris water maze spatial resolution test 1 hour after perfusion on the 10m, 11m, and 12m days after administration. Rats also underwent passive step-down avoidance test 1 hour after reperfusion on the 13m and 14m days after administration. Thirty minutes prior to experimentation, rats in the remaining three groups were intraperitoneally （i.p） injected with 2 mg/kg scopolamine, and rats in the control group were i.p. injected with 2 mL/kg saline. ② Scopolamine-induced learning disorder and memory acquired disorder in mice： Fifty mice were randomly divided into control group, model group, combination group （400 mg/kg GSL ＋200 mg/kg choline）, GSL （400 mg/kg） group, and choline （200 mg/kg） group, with 10 mice/group. Mice were perfused and administrated in the morning, once a day for 9 successive days. Mice in the control group and model group were perfused with 20 mL/kg distilled water and underwent passive step down avoidance test 1 hour after reperfusion on the 8th and 9th day after administration. Twenty minutes prior to training, mice in the remaining three groups were i.p. injected with 2 mg/kg scopolamine, and mice in the control group were i.p. injected with 10 mL/kg saline. ③ Sodium nitrite-induced memory consolidation disorder in mice： Grouping, administration, and testing were the same as mentioned above. After training, mice in the remaining three groups were immediately subcutaneously injected with 120 mg/kg sodium nitrite, and mice in the control group were subcutaneously injected with 20 mL/kg saline. ④ Ethanol-induced memory reconsolidation disorder in mice： Grouping, administration, and testing were the same as mentioned above. At 24 hours after training and 20 minutes before retraining, mice in the remaining four groups were perfused with 10 mL/kg ethanol （0.3 volume fraction）, and mice in the control group were perfused with 10 mL/kg saline. MAIN OUTCOME MEASURES： Synergistic effects of GSL and choline on learning and memory deficits induced by scopolamine, sodium nitrite, and ethanol in experimental animals. RESULTS： All 40 rats and 150 mice were included in the final analysis. ① Synergistic effects of GSL and choline on learning and memory disorders induced by scopolamine in rats： During passive step-down avoidance and Morris water maze spatial resolution tests, the number of error responses and length of maze training in the model group were significantly greater than in the control group （P 〈 0.01）; while the number of error responses and length of maze training in the combination group were significantly less than in the model group, GSL group, and choline group （P 〈 0.05-0.01）. The Q value was 〉 1 after combining administration, which suggests that the combination of GSL and choline had synergistic effects. ② Synergistic effects of GSL and choline on learning disorder and memory-acquired disorder induced by scopolamine in mice： During passive step-down avoidance test, the number of error responses in the model group were significantly greater than in the control group （P 〈 0.01 ）; while the number of error responses in the combination group were significantly less than in the model group, GSL group, and choline group （P 〈 0.05-0.01）. The Q value was 〉 1 after combining administration, which suggests GSL and choline had synergistic effects. ③ Synergistic effects of GSL and choline on memory sodium nitrate-induced consolidation disorder in mice： During passive step down avoidance test, the number of error responses in the model group were significantly less than in the control group （P 〈 0.01 ）; while the number of error responses in the combination group were significantly less than in the model group, GSL group, and choline group （P 〈 0.05-0.01）. The Q value was 〉 1 after combined administration, which suggests GSL and choline had synergistic effects. ④ Synergistic effects of GSL and choline on ethanol-induced memory reconsolidation disorder in mice： During passive step down avoidance test, the number of error responses in the model group were significantly greater than in the control group （P 〈 0.01）; while the number of error responses in the combination group were significantly less than in the model group, GSL group, and choline group （P 〈 0.05-0.01）. The Q value was 〉 1 after combined administration, which suggests GSL and choline had synergistic effects. CONCLUSION： GSL and choline have synergistic effects on learning and memory functions.

Effect of Nao Yikang on choline acetyltransferase and caspase-3 brain expression in a rat model of Alzheimer's disease

BACKGROUND： The main components of the traditional Chinese medicine compound Nao Yikang have been shown to possibly alleviate neural damage. OBJECTIVE： To observe the effects of Nao Yikang on expression of choline acetyltransferase （CHAT） and caspase-3 in the rat brains of an experimental Alzheimer＇s disease （AD） model, and to investigate the mechanisms of potential neuroprotective effects. DESIGN, TIME AND SETTING： A randomized, controlled experiment was performed at the Department of Pathophysiology, Medical School of Nantong University between November 2006 and December 2007. MATERIALS： The main active components of Nao Yikang were as follows： prepared polygonum multiflorum, Rhizoma anemarrhenae, and Rhizoma acori tatarinowii. Nao Yikang granules were prepared by Nantong Hospital of Traditional Chinese Medicine. Ibotenic acid （IBO） was purchased from Sigma-Aldrich, USA, ChAT goat anti-rat antibody from Chemicon, USA, and cleaved caspase-3 rabbit anti-rat （Asp175） （5A1） antibody from Cell Signaling, USA. METHODS： A total of 60 male, Sprague Dawley rats （2 months old） were randomly assigned to 6 groups： sham-surgery, model, Nao Yikang 1.73, 3.45, 6.90 g/kg per day, and piracetam, with 10 rats in each group. Bilateral infusions of 5 pg IBO into the nucleus basalis of Meynert were performed with Hamilton syringe and stereotaxic apparatus for AD model establishment. For the sham-surgery group, rats received 1 μL saline in the identical stereotaxic position. From the second day, Nao Yikang groups were administrated 1.73, 3.45, and 6.90 g/kg per day Nao Yikang, respectively, while the piracetam group received 0.04 g/mL piracetam, the model group received 0.5% sodium carboxymethyl cellulose, and the sham-surgery group received normal saline. Rats were intragastrically administered 1 mL/100 g daily for 28 consecutive days. MAIN OUTCOME MEASURES： Following treatment of the various solutions for 28 days, Western blot was utilized to observe ChAT expression in the frontal cortex of AD rats, and immunohistochemistry was applied to quantify caspase-3-positive cells in the frontal cortex. RESULTS： ChAT protein expression significantly decreased in the model group （P 〈 0.01）, however caspase-3 expression was significantly elevated （P 〈 0.01） compared with the sham-surgery group. Compared with the model group, ChAT protein expression increased in the Nao Yikang 1.73 g/kg per day, 3.45 g/kg per day, 6.90 g/kg per day groups, and the piracetam group （P 〈 0.05 or P 〈 0.01） and the number of caspase-3-positive cells decreased in the Nao Yikang 3.45 g/kg per day and 6.90 g/kg per day groups （P 〈 0.01）. However, there was no change in the number of caspase-3-positive cells in the 3.45 g/kg per day group. CONCLUSION： The traditional Chinese medicine compound Nao Yikang increased ChAT protein expression and suppressed caspase-3 expression in the frontal cortex in a dose-dependent manner.

Preparation of Sn-β-zeolite via immobilization of Sn/choline chloride complex for glucose-fructose isomerization reaction

Well dispersion of tin species in an isolated form is a quite challenge since tin salts are easily hydrolyzed into（hydr）oxides during aqueous stannation of β-zeolite.In this study,immobilization of tin species on high silica commercial β-zeolite by using SnCl_2/Choline chloride（ChCl） complex followed with calcination provided a convenient way to get well dispersed Sn in β-zeolite in the aqueous condition,which was observed based on electron microscopy images,UV visible spectra and X-ray diffraction pattern.The existence of ChCl facilitated tin species to incorporate into zeolite.（1-2）wt%of Sn loaded β-zeolites exhibited good catalytic activity and high selectivity for glucose-fructose isomerization reaction.

Phosphoryl choline-grafted water-soluble carbon nanotube

Water-soluble property is the precondition of biomedical evaluation and application of carbon nanotube （CNT）. Novel water- soluble CNT was synthesized in this letter by grafting phosphoryl choline （PC） onto multi-wall CNTs. Utilizing FTIR, XPS, TGA and TEM, the title CNTs were characterized and it was found that the target products could facilely dissolve in water. 2007 Tao Zhang. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Solubilities of CO_2, CH_4,H_2,CO and N_2 in choline chloride/urea

Solubilities of CO_2, CH_4, H_2, CO and N_2 in choline chloride/urea(ChCl/Urea) were investigated at temperatures ranging from 308.2 to328.2 K and pressures ranging from 0.6 to 4.6 MPa. The results show that the solubilities of gases increase with increasing pressure and decreasing temperature. The solubility of CO_2 is higher than that of CH_4, H_2, CO and N_2, which indicates that ChCl/Urea may be used as a potential solvent for CO_2 capture from the gas mixture. Solubility of CO_2 in ChCl/Urea was fitted by Non-Random Two-Liquid and Redlich-Kwong(NRTL-RK) model, and solubility of CH_4, H_2, CO or N_2 in ChCl/Urea was fitted by Henry's Law. The standard enthalpy, standard Gibbs energy and standard entropy of gases were calculated. Additionally, the CO_2/CH_4 selectivities in water, dry ChCl/Urea and aqueous ChCl/Urea were further discussed.