Limonitum Ameliorates Castor Oil-Induced Diarrhoea in Mice by Modulating Gut Microbiota.

Diarrhoea is a common clinical condition; its pathogenesis is strongly associated with gut microbiota dysbiosis. Limonitum is a well-known traditional Chinese medicine that exerts appreciable benefits regarding the amelioration of diarrhoea. However, the mechanism through which Limonitum ameliorates diarrhoea remains unclear. Here, the efficacy and underlying mechanism of Limonitum decoction (LD) regarding diarrhoea were explored from the aspect of gut microbiota. Castor oil (CO) was used to induce diarrhoea in mice, which were then used to evaluate the effects of LD regarding the timing of the first defecation, diarrhoea stool rate, degree of diarrhoea, diarrhoea score, intestinal propulsive rate, and weight of intestinal contents. The concentrations of short-chain fatty acids (SCFAs), including acetic, propionic, isobutyric, butyric and valeric acids, were analysed by gas chromatography-mass spectrometry (GC-MS). The 16S rRNA high-throughput sequencing technology was applied to evaluate changes in the gut microbiota under exposure to LD. LD was found to effectively ameliorate the symptoms of diarrhoea, and the diversity and relative abundance of gut microbiota were restored to normal levels following LD treatment. Additionally, LD significantly restored the observed reductions in SCFAs. These results provide strong evidence that LD can sufficiently ameliorate diarrhoea in mice by regulating their gut microbiota. The findings presented here highlight that Limonitum may constitute a prospective remedy for diarrhoea.

[The relation between cochlear distortion products and frequency tuning characteristics revealed by laser interferometery].

Objective:To explore the relation between distortion products and frequency tuning charateristics in cochlear basilar membrane.Method:Time variances curves of distortion products and frequency tuning curves of basilar membrane response were measured by laser interferometery in situ guinea pig cochlea.Result:Response sensitivity and tuning sharpness of basilar membrane response decreased with distortion product amplitude and become most low and flat when distortion products disappeared.Conclusion:Distortion products in cochlea are related with hearing sensitivity and frequency tuning. They are indicator whether function of cochlear hearing sensitivity enhancing and frequency tuning sharpening is still at work.

Effect of flaxseed on the fatty acid profile of egg yolk and antioxidant status of their neonatal offspring in Huoyan geese.

The aim of this study was to evaluate the effects of geese's maternal diet supplemented with flaxseed on the fatty acid profiles of egg yolks and the antioxidant status of their offspring. A total of 288 female Huoyan geese (42 weeks old) were randomly allotted to four experimental groups in this 56-day experiment and fed on diets containing flaxseed at 0% (control), 5%, 10% and 15%, respectively. There were nine replicate pens per treatment, with eight geese per replicate pen. The concentration of α-linolenic acid (linear, P<0.01), EPA (20:5n-3; linear, P<0.01), DHA (22:6n-3; quadratic, P=0.03) and n-3 polyunsaturated fatty acid (PUFA) (linear, P<0.01) levels in the yolk lipids increased with increasing dietary flaxseed levels. Yolk palmitic acid (16:0, linear, P=0.05), saturated fatty acid (linear, P=0.04) level and total n-6/n-3 ratio (P<0.01) decreased in a linear fashion as dietary flaxseed levels increased. Increasing dietary flaxseed levels linearly decreased (P=0.01) the total cholesterol in egg yolks. After hatching, three 1-day-old gosling were selected randomly from each replicate to determine blood characteristics and liver antioxidant status. Aspartate aminotransferase activity (linear, P=0.03), total triglycerides (linear, P=0.02) and total cholesterol (linear, P=0.05) contents in blood linearly decreased as the levels of flaxseed increased. A linear dose response to maternal dietary flaxseed was detected for the activities of the goslings' liver enzymes catalase (linear, P=0.01), superoxide dismutase (linear, P<0.01) and glutathione peroxidase (linear, P<0.01). The malondialdehyde (quadratic, P=0.03) and alkaline phosphatase content in the livers of goslings decreased as flaxseed supplementation levels increased. In conclusion, the dietary addition of flaxseed up to 15%, in the maternal diet resulted in increased n-3 PUFA levels in egg yolks and improved the antioxidant status of offspring in a dose-dependent manner.

The effective method for investigation meridian tropism theory in rats.

This present work describes an effective new method for study traditional Chinese medicine (TCM) on meridian tropism (MT) theory, which plays an essential role in clinical selection of TCM according to syndromes and strengthens the therapeutic effects. The new thread included material basis foundation and its tissue distribution study. Xiheliu, the most popular TCM on heart tropism, was investigated by simple and accurate high performance liquid chromatography (HPLC) method. The analysis of plasma after oral administration the total flavonoid of Xiheliu (TFX) exhibited that tamarixetin and kaempferide had the highest concentration and approximately the highest level within 25 min. The mixture of them could last accelerating the urine excretion more than 7 h after a single dose and could not cause the disorder of ion in rats, which was observed in diuretic activity experiment. In view of the reported biological activities was consistent with the effects of Xiheliu, tamarixetin and kaempferide were likely to be the material basis of it. Tissue distribution study showed that the highest level of analytes was in heart, lung, kidney and liver, and most tissues reached maximum level at 30 min post-dose. Since liver was the most important blood-supply tissue, the result of this experiment was in accordance with the MT record of Xiheliu and confirmed that tamarixetin and kaempferide was the material bases of it on MT. This is the first report for the illumination of material basis and the mechanism of Xiheliu on MT by analysis the record of Xiheliu in Compendium of Materia Medica and experimental study.

Removal of ammonium from rare-earth wastewater using natural brucite as a magnesium source of struvite precipitation.

This paper presents a study regarding ammonium removal from rare-earth wastewater by struvite precipitation with natural brucite mineral as a source of magnesium. Experimental results indicated that a pH ranging from 8.5 to 9.5 was the optimum for the removal of ammonium using the soluble form of brucite as a magnesium source. Additionally, when solid brucite was used as a magnesium source as well as an alkali reagent, the initial ammonium concentration of 4,535 mg/L decreased to 239-317 mg/L after an reaction time of 12 h in wastewater treated with the S/L (solid brucite/liquid wastewater) ratios ranging from 31.2 to 63.2 g/L. Furthermore, as some non-reacted brucite still remained in the precipitates obtained at the end of reaction, the precipitates were subjected to reuse. The reuse results demonstrated that the reuse of the precipitates obtained with 63.2 g/L was feasible, and almost half of the brucite dose could be saved.

Masting in ponderosa pine: comparisons of pollen and seed over space and time.

Many plant species exhibit variable and synchronized reproduction, or masting, but less is known of the spatial scale of synchrony, effects of climate, or differences between patterns of pollen and seed production. We monitored pollen and seed cone production for seven Pinus ponderosa populations (607 trees) separated by up to 28 km and 1,350 m in elevation in Boulder County, Colorado, USA for periods of 4-31 years for a mean per site of 8.7 years for pollen and 12.1 for seed cone production. We also analyzed climate data and a published dataset on 21 years of seed production for an eighth population (Manitou) 100 km away. Individual trees showed high inter-annual variation in reproduction. Synchrony was high within populations, but quickly became asynchronous among populations with a combination of increasing distance and elevational difference. Inter-annual variation in temperature and precipitation had differing influences on seed production for Boulder County and Manitou. We speculate that geographically variable effects of climate on reproduction arise from environmental heterogeneity and population genetic differentiation, which in turn result in localized synchrony. Although individual pines produce pollen and seed, only one-third of the covariation within trees was shared. As compared to seed cones, pollen had lower inter-annual variation at the level of the individual tree and was more synchronous. However, pollen and seed production were similar with respect to inter-annual variation at the population level, spatial scales of synchrony and associations with climate. Our results show that strong masting can occur at a localized scale, and that reproductive patterns can differ between pollen and seed cone production in a hermaphroditic plant.

Complete 1H and 13C assignments of 8-C-beta-D-[2-O-(E)-p-coumaroyl]glucopyranosyl-2-(2-hydroxy)propyl-7-methoxy-5-methylchromone.

1H and 13C NMR spectra of 8-C-beta-D-[2-O-(E)-p-coumaroyl] glucopyranosyl-2-(2-hydroxy)propyl-7-methoxy-5-methylchromone were completely assigned by 2D NMR observations. Especially the 1H assignments of the glucosyl and hydroxyl protons were achieved by utilizing HMQC, HMBC, 1H-1H COSY and DEPT techniques together with a heavy water exchange 1H NMR experiment.

Regulation of rat carboxylesterase expression by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD): a dose-dependent decrease in mRNA levels but a biphasic change in protein levels and activity.

Carboxylesterases play an important role in the metabolism of endogenous lipids and foreign compounds; therefore, xenobiotic regulation of carboxylesterase gene expression has both physiological and pharmacological significance. We previously reported that beta-naphthoflavone and 3-methylcholanthrene, two potent inducers for cytochrome P4501A enzymes, had opposing effects on the expression of hydrolase S, a secretory carboxylesterase. Beta-naphthoflavone caused suppression, whereas 3-methylcholanthrene caused induction of the expression of this enzyme. The aim of the present study was to determine the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), another prototypical cytochrome P4501A inducer, on the expression of this and several other rat carboxylesterases (hydrolases A, B, and C) in liver and extrahepatic tissues. Rats received TCDD treatment at nonlethal (

Calpain cleavage promotes talin binding to the beta 3 integrin cytoplasmic domain.

Talin links integrin beta cytoplasmic domains to the actin cytoskeleton and is involved in the clustering and activation of these receptors. To understand how talin recognizes integrin beta cytoplasmic domains, we configured surface plasmon resonance methodology to measure the interaction of talin with the beta3 integrin cytoplasmic domain. Here we report that the N-terminal approximately 47-kDa talin head domain (talin-H) has a 6-fold higher binding affinity than intact talin for the beta3 tail. The affinity difference is mainly due to a difference in k(on). Calpain cleavage of intact talin released talin-H and resulted in a 16-fold increase in apparent K(a) and a 100-fold increase in apparent k(on). The increase in talin binding after cleavage was greater than predicted for stoichiometric liberation of free talin-H. This additional increase in binding was due to cooperative binding of talin-H and talin rod domain to the beta3 tail. Talin resembles ERM (ezrin, radixin, moesin) proteins in possessing an N-terminal FERM (band four-point-one, ezrin, radixin, moesin) domain. These data show that the talin FERM domain, like that in the ERM proteins, is masked in the intact molecule. Furthermore, they suggest that talin cleavage by calpain may contribute to the effects of the protease on the clustering and activation of integrins.

Transcriptional regulation of the human acid alpha-glucosidase gene. Identification of a repressor element and its transcription factors Hes-1 and YY1.

Acid alpha-glucosidase, the product of a housekeeping gene, is a lysosomal enzyme that degrades glycogen. A deficiency of this enzyme is responsible for a recessively inherited myopathy and cardiomyopathy, glycogenesis type II. We have previously demonstrated that the human acid alpha-glucosidase gene expression is regulated by a silencer within intron 1, which is located in the 5'-untranslated region. In this study, we have used deletion analysis, electrophoretic mobility shift assay, and footprint analysis to further localize the silencer to a 25-base pair element. The repressive effect on the TK promoter was about 50% in both orientations in expression plasmid, and two transcriptional factors were identified with antibodies binding specifically to the element. Mutagenesis and functional analyses of the element demonstrated that the mammalian homologue 1 of Drosophila hairy and Enhancer of split (Hes-1) binding to an E box (CACGCG) and global transcription factor-YY1 binding to its core site function as a transcriptional repressor. Furthermore, the overexpression of Hes-1 significantly enhanced the repressive effect of the silencer element. The data should be helpful in understanding the expression and regulation of the human acid alpha-glucosidase gene as well as other lysosomal enzyme genes.

Biosynthesis, molecular structure, and domain architecture of potato suberin: a (13)C NMR study using isotopically labeled precursors.

Although suberin in potato wound periderm is known to be a polyester containing long-chain fatty acids and phenolics embedded within the cell wall, many aspects of its molecular structure and polymer-polymer connectivities remain elusive. The present work combines biosynthetic incorporation of site-specifically (13)C-enriched acetates and phenylalanines with one- and two-dimensional solid-state (13)C NMR spectroscopic methods to monitor the developing suberin polymer. Exogenous acetate is found to be incorporated preferentially at the carboxyl end of the aliphatic carbon chains, suggesting addition during the later elongation steps of fatty acid synthesis. Carboxyl-labeled phenylalanine precursors provide evidence for the concurrent development of phenolic esters and of monolignols typical of lignin. Experiments with ring-labeled phenylalanine precursors demonstrate a predominance of sinapyl and guaiacyl structures among suberin's phenolic moieties. Finally, the analysis of spin-exchange (solid-state NOESY) NMR experiments in ring-labeled suberin indicates distances of no more than 0.5 nm between pairs of phenolic and oxymethine carbons, which are attributed to the aromatic-aliphatic polyester and the cell wall polysaccharide matrix, respectively. These results offer direct and detailed molecular information regarding the insoluble intermediates of suberin biosynthesis, indicate probable covalent linkages between moieties of its polyester and polysaccharide domains, and yield a clearer overall picture of this agriculturally important protective material.

NMR studies of structure and dynamics in fruit cuticle polyesters.

Cutin and suberin are support polymers involved in waterproofing the leaves and fruits of higher plants, regulating the flow of nutrients among various plant organs, and minimizing the deleterious impact of microbial pathogens. Despite the complexity and intractable nature of these plant biopolyesters, their molecular structure and development are amenable to study by suitable solid-state and solution-state NMR techniques. Interactions of tomato cutin with water were examined by solid-state 2H and 13C NMR, showing that water films enhance rapid segmental motions of the acyl chains and are associated with a fivefold increase in surface elasticity upon cutin hydration. The suberization of wounded potato tissues was studied by solid-state 13C NMR, revealing the likely phenylpropanoid structures that permit dense cross-linking of the suberin structure and their proximity to the cell-wall polysaccharides. Finally, two new approaches were developed to elucidate the molecular structures of these biopolymers: partial depolymerization followed by spectroscopic analysis of the soluble oligomers; and swelling of the intact materials followed by magic-angle spinning (MAS) NMR analysis.

Improvement of early postburn cardiac function by use of Panax notoginseng and immediate total eschar excision in one operation.

Cardiac dysfunction development in the early stage postburn has been an important problem in burn treatment. However, no effective therapies are available for use in clinical practice. In this study, we sought to determine whether early total eschar excision (EEE) in one operation and the traditional Chinese herb Panax notoginseng (PNS) would be helpful in improving early postburn cardiac function. 160 Wistar rats were randomly divided into burn (burn group, n = 50), burn treated with EEE (EEE group, n = 50), burn treated with PNS (PNS group, n = 50) groups and normal controls (n = 10). All rats except the normal control were given a 30% TBSA full skin thickness burn and resuscitated with Ringer's lactate. EEE was performed immediately after the burn group received the first intraperitoneal injection of Ringer's lactate. The wound was covered with homoskin from normal rats. In the PNS group, two doses of PNS (200 mg/kg for each dose) were given intraperitoneally immediately and 4 h postburn. Cardiac contractile function and cardiac troponin T (TnT) were determined at 1, 3, 6, 12 and 24 h postburn. Results showed that cardiac contractile parameters including AOSP, AODP, LVSP and +dp/dt(max) all declined and were still significantly lower than the control values at 24 h postburn. Cardiac TnT was elevated markedly and reached a level 25 times higher than control at 12 h postburn. In EEE and PNS groups, the reduction of cardiac contractile function was limited as compared with that in the burn group. Levels of TnT in both EEE and PNS groups were significantly lower than in the burn group 6 h postburn later. The findings of this study demonstrated that both EEE and PNS were effective in improving early postburn cardiac function.

[Isolation and identification of a saponine from Patrinia scabiosaefolia].

A saponine compound was isolated from the acetone extract of the roots and rhizomes of Patrinia scabiosaefolia Fish. ex Link. Its structure was identified by combination of chemical reaction and spectrum analysis as 3-O-alpha-L-rhamnopyranosyl-(1-2)-beta-D-xylopyranosyl oleanolic acid (giganteaside D), and part of its 13C-NMR data was revised by the 2D-NMR. The compound was found in the Patrinia for the first time.

[Effect of Cladonia alpestris on Trichomonas vaginalis in vitro].

In this paper, an experimental research is reported on the effect of water extract of Cladonia alpestris and S-(-) usnic acid on Trichomonas vaginalis in vitro. The results showed that both the water extract of Cladonia alpestris and S-(-)usnic acid exhibited a strong effect against Trichomonas vaginalis in vitro. As the time of action of the agents was prolonged, the mortality of Trichomonas vaginalis increased. For S-(-) usnic acid, 0.4 mg/ml was the lowest effective concentration against Trichomonas vaginalis in vitro. No remarkable differences were found in the effect of the S-(-)usnic acid and metronidazole at concentrations of 0.4 mg/ml and 0.6 mg/ml.

[Therapeutic effects of anti-rheumatic tablet in ankylosing spondylitis].

188 patients of ankylosing spondylitis were treated with anti-rheumatic tablet compared with 30 patients with Indomethacin as control group. The results showed that spinal antiflexion and finger-ground test of both treated group and control group had improved significantly (P less than 0.05), but lateral curvature movement, thorax expansion test, 20 m walking time, and the levels of IgG, IgA, IgM, C3, ESR, CRP of the treated group had marked difference compared with those treated before (P less than 0.01). It was proved that in the treated group, the marked effective rate was 53.72%, while in the control group was 20.00%. There was significant difference between the two groups in effective rate (P less than 0.001). This revealed that anti-rheumatic tablet is a kind of ideal drug in treating ankylosing spondylitis.

Studies of safe maximal daily dietary Se-intake in a seleniferous area in China. Part II: Relation between Se-intake and the manifestation of clinical signs and certain biochemical alterations in blood and urine.

Selenosis occurs in areas of Enshi county because of the high Se content of the food. Morphological changes in finger-nails were used as the main criterion for clinical diagnosis of selenosis. Pathological nails were observed to occur almost only in adults, not at all in young children and very seldom in teenagers. Symptoms of selenosis in susceptible patients were found at or above an Se-intake of 910 micrograms/d, corresponding to a blood Se level of 1.05 mg/L. There was no evidence for an increased susceptibility to dental caries due to high Se consumption, and an increase in Se-intake seems unlikely to reduce the beneficial effects of fluoride on caries. No abnormalities of liver or heart were seen by supersonic B or electrocardiographic examinations. The biochemical investigations showed that with increasing whole blood Se the ratio of plasma Se to erythrocyte Se tended to decrease. As Se-intake increases to over 750 micrograms daily, the ratio decreases to near a minimal level. Reduced glutathione in whole blood decreases within a blood Se range of 1.01 to 2.28 micrograms in the high Se area. The amount of trimethylselenonium ion excreted in urine increased with the increase of urinary Se. Cases with prolonged prothrombin time occurred as blood Se increased to a level above 1 mg/L. The white blood cell count also increased significantly. Quantitative values were obtained only for ratio of plasma-Se to erythrocyte-Se for prothrombin time and for maintenance of nail Symptoms of susceptible patients. The overall results indicated that a daily Se-intake of 750-850 micrograms [corrected] might be the marginal level of safe intake. When other variable factors are also taken into consideration a daily Se-intake of 400 micrograms [corrected] is suggested as the maximum daily safe intake. At this level of Se-intake the corresponding approximate tissue Se levels are: whole blood 0.559 mg/L, plasma 0.327 mg/L, urine excretion 173 micrograms/d, hair 3.60 mg/kg, toe-nails 4.25 mg/kg, and finger-nails 4.70 mg/kg.

Studies of safe maximal daily dietary selenium intake in a seleniferous area in China. I. Selenium intake and tissue selenium levels of the inhabitants.

Studies of marginal safe Se-intake have been carried out in a seleniferous section of China since 1985. Three areas with low, medium and high Se levels were selected for this study. The respective average daily Se-intake (mean +/- SE) was 70.5 +/- 4.8 micrograms, 194.7 +/- 22.9 micrograms and 1438.2 +/- 76.3 micrograms for males, and 62.0 +/- 3.6 micrograms, 198.1 +/- 23.8 micrograms and 1238.5 +/- 64.6 micrograms for females (average body weight: male 55 Kg, female 53 Kg). When the increasing rate of Se-intake was compared with the corresponding tissue-Se levels it was found that the whole blood Se-level reflected more closely the physiological range of Se-intake, while at higher Se-intakes it became less sensitive than the levels in hair, finger-nail and toe-nail, which were comparable to the sensitivity of urine. It is suggested that hair, finger- and toe-nail may all act as excretory organs when excess amounts of Se are ingested. Hair- and blood-Cd are somewhat higher in residents of the high Se area, but whether they have influenced human Se-metabolism at the high level of Se-intake is not yet known. Significant correlations on log-log plots were obtained between levels of daily Se-intake and whole blood r = 0.878), breast milk (r = 0.899) and 24-h-urine (r = 0.859). Highly significant correlations on log-log plots between levels of tissue were also obtained: urine Se--plasma Se (r = 0.968), whole blood Se--hair Se (r = 0.952), fingernail Se--toenail Se (r = 0.919), hair Se--fingernail Se (r = 0.914), hair Se--toenail Se (r = 0.891), whole blood Se--toenail Se (r = 0.849) and whole blood Se--fingernail Se (r = 0.836). The highly significant correlations found between the Se-intake and the tissue-Se level, and also between the Se levels of various tissues, could possibly conveniently be used to convert the known tissue-Se level to the corresponding Se-intake. Taken together with the wide range of Se-intakes and corresponding tissue-Se levels this would provide the necessary conditions for studying the marginal and maximal safe Se-intakes in humans.