Co-exposure to fluoride and sulfur dioxide induces abnormal enamel mineralization in rats via the FGF9-mediated MAPK signaling pathway.

Fluoride (F) and sulfur dioxide (SO2) contamination is recognized as a public health concern worldwide. Our previous research has shown that Co-exposure to F and SO2 can cause abnormal enamel mineralization. Ameloblastin (AMBN) plays a crucial role in the process of enamel mineralization. However, the process by which simultaneous exposure to F and SO2 influences enamel formation by regulating AMBN expression still needs to be understood. This study aimed to establish in vivo and in vitro models of F-SO2 Co-exposure and investigate the relationship between AMBN and abnormal enamel mineralization. By overexpressing/knocking out the Fibroblast Growth Factor 9 (FGF9) gene, we investigated the impact of FGF9-mediated Mitogen-Activated Protein Kinase (MAPK) signaling on AMBN synthesis to elucidate the mechanism underlying the induction of abnormal enamel mineralization by F-SO2 Co-exposure in rats. The results showed that F-SO2 exposure damaged the structure of rat enamel and ameloblasts. When exposed to F or SO2, gradual increases in the protein expression of FGF9 and phosphorylated p38 mitogen-activated protein kinase (p-P38) were observed. Conversely, the protein levels of AMBN, phosphorylated extracellular signal-regulated kinase (p-ERK), and phosphorylated c-Jun N-terminal kinase (p-JNK) were decreased. AMBN expression was significantly correlated with FGF9, p-ERK, and p-JNK expression in ameloblasts. Interestingly, FGF9 overexpression reduced the levels of p-ERK and p-JNK, worsening the inhibitory effect of F-SO2 on AMBN. Conversely, FGF9 knockout increased the phosphorylation of ERK and JNK, partially reversing the F-SO2-induced downregulation of AMBN. Taken together, these findings strongly demonstrate that FGF9 plays a critical role in F-SO2-induced abnormal enamel mineralization by regulating AMBN synthesis through the JNK and ERK pathways.

The role of FGF9-mediated TGF-ß1/Smad signaling in enamel hypoplasia induced by exposure to fluoride and SO2 in rats.

Many geographical areas of the world are polluted by both fluoride and sulfur dioxide (SO2). However, the effects of simultaneous exposure to fluoride and SO2 on teeth are unknown. Fibroblast growth factor-9 (FGF9) and transforming growth factor-ß1 (TGF-ß1) are key signaling molecules in enamel development. The purpose of the study was to explore the effects of co-exposure to fluoride and sulfur dioxide on enamel and to investigate the role and mechanism of FGF9 and TGF-ß1. First, sodium fluoride (NaF) and SO2 derivatives were used to construct rat models and evaluate the enamel development of rats. Then, TGF-ß1 (cytokine) treatment, SIS3 (inhibitor) treatment and FGF9 gene knockdown were used to explore the mechanism of enamel damage in vitro. The results showed that enamel column crystals in the exposed group were characterized by enamel hypoplasia, as indicated by alterations such as disarrangement of enamel column crystals, space widening and breakage. Ameloblasts also showed pathological changes such as ribosome loss, mitochondrial swelling, nuclear fragmentation and chromatin aggregation. The protein expression of FGF9 was higher and the protein expression of AMBN, TGF-ß1 and p-Smad2/3 protein was lower in the groups treated with fluoride and SO2 individually or in combination compared with the control group. Further studies showed that TGF-ß1 significantly upregulated p-Smad2/3 and AMBN protein expression and reduced the inhibitory effects of fluoride and SO2; furthermore, SISI blocked the effect of TGF-ß1. In addition, knockdown of FGF9 upregulated TGF-ß1 protein expression, further activated Smad2/3 phosphorylation, eliminated the inhibitory effects of fluoride and SO2, and increased the protein expression of AMBN. In brief, the study confirms that co-exposure to fluoride and SO2 can result in enamel hypoplasia in rats and indicates that the underlying mechanism may be closely related to the effect of FGF9 on enamel matrix protein secretion through inhibition of the TGF-ß1/Smad signaling pathway.

Transfer and transformation characteristics of Zn and Cd in soil-rotation plant (Brassica napus L and Oryza sativa L) system and its influencing factors.

Rice-rape rotation is a widely practiced cropping system in China. However, changes in soil properties and management could change the bioavailability of Cd, In order to explore the occurrence state, transportation and transformation characteristics of heavy metals Cd and Zn in rice-rape rotation system in Guizhou karst area with high background value of Cd. In the karst rice-rape rotation area, the physical and chemical properties of soil, chemical specifications and activities of Cd and Zn at different soil depths and during various crop growth stages, and the bioaccumulation of Cd and Zn in different tissues of rice and rape were studied by field experiment and laboratory analysis. The bioaccumulation of Cd and Zn and the effects of physical and chemical soil properties on the activities and bioavailabilities of Cd and Zn during rice-rape rotation were explored. The findings revealed that soil particle size, composition, pH, redox potential, soil organic matter, and Cd and Zn contents varied dramatically, especially in deep soils. The physical and chemical properties of the deep and surface soils were significantly related to the bioaccumulation of Cd and Zn. Cd and Zn are activated during crop rotation. Cd was easier to be enriched in rice, while Zn was easier to be enriched in rape. The correlation between Cd and Zn contents in Brassica campestris L and their enrichment abilities were not significant, but that in Oryza sativa L were significant. During rice-rape rotation, the chemical speciations and activities of Cd and Zn changed with the changes of soil properties and waterlogging environment. This study had important basic guiding significance for the evaluation, prevention and control of heavy metal pollution, and improving soil quality in different rotation systems in karst areas, and was conducive to promoting the safe production of rape and rice.

Sodium Fluoride and Sulfur Dioxide Derivatives Induce TGF-ß1-Mediated NBCe1 Downregulation Causing Acid-Base Disorder of LS8 Cells.

The aim of the present work was to assess whether the combination of sodium fluoride (NaF) and sulfur dioxide derivatives (SO2 derivatives) affects the expression of the electrogenic sodium bicarbonate cotransporter NBCe1 (SLC4A4), triggering an acid-base imbalance during enamel development, leading to enamel damage. LS8 cells was taken as the research objects and fluorescent probes, quantitative real-time polymerase chain reaction (qRT-PCR), western blot, and factorial analysis were used to clarify the nature of the fluoro-sulfur interaction and the potential signaling pathway involved in the regulation of NBCe1. The results showed that exposure to fluoride or SO2 derivatives resulted in an acid-base imbalance, and these changes were accompanied by inhibited expression of NBCe1 and TGF-ß1; these effects were more significant after fluoride exposure as compared to exposure to SO2 derivatives. Interestingly, in most cases, the toxic effects during combined exposure were significantly reduced compared to the effects observed with fluoride or sulfur dioxide derivatives alone. The results also indicated that activation of TGF-ß1 signaling significantly upregulated the expression of NBCe1, and this effect was suppressed after the Smad, ERK, and JNK signals were blocked. Furthermore, fluoride and SO2 derivative-dependent NBCe1 regulation was found to require TGF-ß1. In conclusion, this study indicates that the combined effect of fluorine and sulfur on LS8 cells is mainly antagonistic. TGF-ß1 may regulate NBCe1 and may participate in the occurrence of dental fluorosis through the classic TGF-ß1/Smad pathway and the unconventional ERK and JNK pathways.

Determination of cadmium in Chinese pepper and its health implications based on bioaccessibility.

The contamination of cadmium (Cd) in agro-products causes major concern because of its potential dietary risks. In this study, a total of 647 pepper samples from 21 provinces in China were randomly collected according to the distribution of pepper production. Cd pollution levels in Chinses pepper and its health risks were evaluated based on bioaccessibility, which was measured by the physiologically based extraction test (PBET). The results showed that Cd concentration in all pepper ranged from 0.002 to 1.470 mg/kg, with an average of 0.222 mg/kg and a median of 0.132 mg/kg. The highest daily intake of Cd was observed in the female child group (4.037 × 10-5 mg/kg bw/day), which accounted for 4% of the maximum daily permissible dose - 0.001 mg/kg bw/day. The target hazard quotients of Cd were all lower than 1, indicating low potential non-carcinogenic health risks to residents via the consumption of pepper. Notably, carcinogenic risk values suggested potential adverse health effects to adults, while after considering the bioaccessibility of Cd in pepper (mean of 43.07%), those values had fallen under the acceptable level (1 × 10-4). This may indicate that dietary risk assessment of heavy metals in crops could not be conducted just based on their content; the bioaccessibility of metals is also an important factor for consideration.

Arbuscular mycorrhizal fungi improve the growth and drought tolerance of Cinnamomum migao by enhancing physio-biochemical responses.

Drought is the main limiting factor for plant growth in karst areas with a fragile ecological environment. Cinnamomum migao H.W. Li is an endemic medicinal woody plant present in the karst areas of southwestern China, and it is endangered due to poor drought tolerance. Arbuscular mycorrhizal fungi (AMF) are known to enhance the drought tolerance of plants. However, few studies have examined the contribution of AMF in improving the drought tolerance of C. migao seedlings. Therefore, we conducted a series of experiments to determine whether a single inoculation and coinoculation of AMF (Claroideoglomus lamellosum and Claroideoglomus etunicatum) enhanced the drought tolerance of C. migao. Furthermore, we compared the effects of single inoculation and coinoculation with different inoculum sizes (20, 40, 60, and 100 g; four replicates per treatment) on mycorrhizal colonization rate, plant growth, photosynthetic parameters, antioxidant enzyme activity, and malondialdehyde (MDA) and osmoregulatory substance contents. The results showed that compared with nonmycorrhizal plants, AMF colonization significantly improved plant growing status; net photosynthetic rate; superoxide dismutase, catalase, and peroxidase activities; and soluble sugar, soluble protein, and proline contents. Furthermore, AMF colonization increased relative water content and reduced MDA content in cells. These combined cumulative effects of AMF symbiosis ultimately enhanced the drought tolerance of seedlings and were closely related to the inoculum size. With an increase in inoculum size, the growth rate and drought tolerance of plants first increased and then decreased. The damage caused by drought stress could be reduced by inoculating 40-60 g of AMF, and the effect of coinoculation was significantly better than that of single inoculation at 60 g of AMF, while the effect was opposite at 40 g of AMF. Additionally, the interaction between AMF and inoculum sizes had a significant effect on drought tolerance. In conclusion, the inoculation of the AMF (Cl. lamellosum and Cl. etunicatum) improved photosynthesis, activated antioxidant enzymes, regulated cell osmotic state, and enhanced the drought tolerance of C. migao, enabling its growth in fragile ecological environments.

Three new acenaphthene derivatives from rhizomes of Musa basjoo and their cytotoxic activity.

Three new acenaphthene derivatives cis-3-(4'-methoxyphenyl)-acenaphthene-1, 2-diol (1), trans-(1S, 2S)-3-phenyl-acenaphthene-1, 2-diol (2) and 8-(4-hydroxyphenyl)-2H-acenaphthylen-1-one (3) in company with six known compounds were isolated from the 70% ethanol extract of the rhizomes of Musa basjoo. Those chemical constituents were separated and purified by macroreticular resin, silica gel, Toyopearl HW-40F, SephadexLH-20 and other chromatographic methods, respectively. The chemical structures of new compounds were elucidated by HR-ESI-MS, 1H NMR, 13C NMR, HMQC, HMBC spectrum and specific optical rotations. Compound 4 was isolated for the first time from the genus Musa and compound 7 was firstly assigned the carbon spectrum. Furthermore, the cytotoxic activity of compounds 1-9 against WM9, MDA-MB231, HeLa, K562, DU145 and PC3 was screened with cisplatin as a positive control. Compound 9 showed promising cytotoxic activities with IC50 values of 2.65 ± 0.38 µM against the HeLa cell lines, while compound 8 possessed significant cytotoxicity with IC50 values of 6.51 ± 0.44, 18.54 ± 0.68 and 7.98 ± 1.44 µM against the HeLa, MDA-MB231 and WM9 cell lines, respectively.

Fluorine enrichment of vegetables and soil around an abandoned aluminium plant and its risk to human health.

In the process of electrolytic aluminium production, a large amount of fluoride is deposited into the surrounding environment. The growth of crops within these areas creates a state of high stress in plants that can easily result in excessive fluorine enrichment in agricultural products, which in turn poses a health threat to humans via the food chain. However, it is not clear what the degree of influence is or how long the impacts of fluorides in soil and agricultural products last for. In this study, 122 vegetable species and 36 surface soil samples were collected within 10 km of an aluminium plant that had been abandoned for five years. The single factor index and the geological accumulation index methods were used to evaluate the degree of fluorine pollution in vegetables and soil within the study area. The hazard index and the hazard quotient were used to assess vegetable and soil health risks, respectively. The results showed that the fluorine content of 89.26% of agricultural products exceeded the Chinese Maximum Levels of Contaminants in Foods and was higher than in other regions, indicating that the emissions of fluoride from the aluminium plant still had a significant effect on fluorine accumulation in vegetables. The total fluorine (Ft) content in the surrounding surface soil (average value 1328.32 mg kg-1) was similar to the soil fluorine background in the nearby uncontaminated settlement area, but the soil water-soluble fluorine (Fw) was very high, indicating that the fluorine pollutants emitted from the aluminium plant had gradually been diluted by the high background of the original soil, but the solubility of these pollutants was higher (average 15.00 mg kg-1) and would continue to threaten the safety of agricultural products in this region. Within 1 km of the abandoned aluminium plant, the degree of fluorine enrichment in vegetables, and Ft and Fw in soil were significantly higher than other ranges, indicating that this was the main fluoride deposit area. Based on the HI and HQ evaluation, vegetables and ground dust around the aluminium plant still had high non-carcinogenic risks for children, even though it has been abandoned for more than five years.

Relationships between soil properties and the accumulation of heavy metals in different Brassica campestris L. growth stages in a Karst mountainous area.

The speciation and activity of heavy metals in farmland were changed with the different soil properties and flooded environment, especially in the complex and rainy environment in soil of Guizhou Province. The objective of this study was to explore the concentrations of a variety of heavy metal activity and the speciation of those heavy metals in rhizosphere soil at different growth stages of Brassica campestris L. in a Karst mountainous area. Tessier's five-stage sequential extraction procedure, the potential ecological risk index, a Bayesian network, accumulation factors, translocation factors and a laboratory simulation experiment were applied in this study. The results showed that (1) no heavy metal concentrations (except the Cd concentration) exceeded the limits of the soil environmental quality risk control standards for soil contamination of agricultural land in China (GB15618-2018). (2) The orders of the accumulation factor and translocation factor values were Zn > Cd > Cu > Pb > Cr and Cd > Cu > Zn > Pb > Cr, respectively. The order of the heavy metal contents of different tissues during the whole growth period was roots > leaves > stems. (3) The indoor simulation test exhibited that the dry-wet alternation and flooding can reduce Cd activity in soil. (4) Redox potential (Eh), rather than pH or organic matter, was the main factor impacting the total content and chemical speciation of heavy metals in the soil, based on a dynamic Bayesian network. Based on the results, we suggest that the activity of heavy metals should be improved by using dry-wet alternation, whereas the proportions of ion-exchangeable forms of heavy metals are relatively low in the study area (except for Cd). Several measures may be taken to enhance soil acidity and reduce the Cd activity during Brassica campestris L. cultivation.

Changes in energy metabolism and macrophage polarization: Potential mechanisms of arsenic-induced lung injury.

Exposure to arsenic is epidemiologically associated with increased lung disease. In detailing the mechanism by which arsenic exposure leads to disease, studies have emphasized that metabolic reprogramming and immune dysfunction are related to arsenic-induced lung injury. However, the association between the mechanisms listed above is not well understood. Thus, the current study aimed to investigate the interaction of energy metabolism and macrophage polarization, by which arsenic exposure adversely induced lung injury in both in vitro and human studies. First, we confirmed a shift to glycolytic metabolism resulting from mitochondrial dysfunction. This shift was accompanied by an increase in the levels of phosphorylated PDHE1α (S293) and PDK1 and a concomitant marked increase in several key markers of the HIF-1α signaling pathway (HIF-1α, p-PKM2, GLUT1 and HK-2). In addition, utilizing an in vitro model in which lung epithelial cells are cultured with macrophages, we determined that arsenic treatment polarizes macrophages towards the M2 phenotype through lactate. In the human study, the serum lactate and TGF-ß levels were higher in arsenic-exposed subjects than that in reference subjects (t= 4.50, 6.24, both p < 0.05), while FVC and FEV1 were both lower (t= 5.47, 7.59, both p < 0.05). Pearson correlation analyses showed a significant negative correlation between the serum TGF-ß and lactate levels and the lung function parameters (pcorrelation<0.05). In mediation analyses, lactate and TGF-ß significantly mediated 24.3% and 9.0%, respectively, of the association between arsenic and FVC (pmediation<0.05), while lactate and TGF-ß significantly mediated 22.2% and 12.5%, respectively, of the association between arsenic and FEV1 (pmediation<0.05). Together, the results of the in vitro and human studies indicated that there is complex communication between metabolic reprogramming and immune dysfunction, resulting in exacerbated effects in a feedback loop with increased arsenic-induced lung damage.

[Shenxiong Glucose Injection inhibits H_2O_2-induced apoptosis of H9c2 cells by activating PI3K/AKT pathway].

The aim of this paper was to explore the mechanism of Shenxiong Glucose Injection antagonizing apoptosis of H9 c2 cells induced by H_2O_2. H9 c2 cells were pretreated with 1. 7%,3. 4% and 6. 8% Shenxiong Glucose Injection,and then H_2O_2 was introduced to induce apoptosis in vitro. Cell viability was detected by MTS assay,morphological changes of apoptosis were observed by AO/EB fluorescence staining,apoptosis rate was detected by Annexin/PI method,cell expression profile was detected by gene chip technology,the mRNA of PIK3 CA,Bcl-2,Bax,caspase-3 and GAPDH were detected by qRT-PCR,the protein expression levels of PIK3 CA,AKT,P-AKT,Bcl-2,Bax and caspase-3 were detected by Western blot,and the contents of LDH and MDA were detected by kit. The results showed that Shenxiong Glucose Injection of different concentrations significantly increased the viability of H9 c2 cells treated with H_2O_2( P<0. 01),and reversed H_2O_2-induced apoptosis( P< 0. 01). The microarray experiments showed that 138 genes were altered in H9 c2 cells after treatment with Shenxiong Glucose Injection. The differential expression fold of PIK3 CA associated with PI3 K/AKT pathway was 3. 59. The results of qRT-PCR and Western blot showed that Shenxiong Glucose Injection could down-regulate the mRNA and protein expression levels of caspase-3( P<0. 01),up-regulate the mRNA and protein expression level of PIK3 CA and Bcl-2( P<0. 01),and up-regulate the phosphorylation levels of AKT( P<0. 01) in H_2O_2-treated H9 c2 cells. The protective effect of Shenxiong Glucose Injection on H_2O_2 cells injury was significantly inhibited by LY294002,a PI3 K/AKT pathway inhibitor. The results suggested that Shenxiong Glucose Injection may inhibit H_2O_2-induced H9 c2 cells apoptosis by regulating PI3 K/AKT signaling pathway.

Design, Synthesis and Anticancer Evaluation of Fangchinoline Derivatives.

Twenty fangchinoline derivatives were synthesized from the natural product fangchinoline, and their anticancer activities on human breast cancer MDA-MB-231 cell line, human prostate cancer PC3 cell line, human melanoma WM9 cell line and human leukaemia HEL and K562 cell lines were evaluated. The biological result showed that those derivatives exhibited potent activities on inhibiting cancer cell growth, and the structure-activity relationships were investigated. Among them, compound 4g, which was protected by benzoyl group in 7-phenolic position and nitrified in 14-position, showed impressive inhibition on all 5 cancer cell lines, especially WM9 cell line, with an IC50 value of 1.07 µM. Further mechanistic studies demonstrated that compound 4g may induce cancer cell death by apoptotic means. These research results suggested that compound 4g could be a lead for the further development toward an anticancer agent against human melanoma WM9 in the future.

[Evaluation and cumulative characteristics of heavy metals in soil-Uncaria rhynchophylla system of different functional areas].

Soil and Uncaria rhynchophylla in different functional areas were selected for the study,the content of heavy metals such as As, Cd, Cu, Cr, Pb, and Hg in soil and U. rhynchophylla was discussed, the characteristics of their accumulation in the U.rhynchophylla was analyzed, the contamination levels of heavy metals in soil in different functional areas was evaluated. The results showed that content of Cu, As, Pb and Cr in soil was being cropland>woodland>wasteland, content of Cd was being woodland>cropland>wasteland, content of Hg was being cropland>woodland>wasteland. According to quality standard of soil environment, soil Cd in woodland, cropland and wasteland all exceeded the state-level standards, soil Cd in woodland exceeded the secondary standard, soil Hg in cropland and wasteland all exceeded the state-level standards. According to technical conditions of green food producing area, soil Cd in woodland exceeded the limit value of standard. According to Green Trade Standards of Importing Exporting Medicinal Plants Preparations,the content of heavy metals of U.rhynchophylla in cropland,woodland and wasteland were correspond to the specification. From the single factor pollution index, the soil in woodland was polluted by Cd. From the comprehensive pollution index, the soils in different functional areas were not contaminated by heavy metals. The enrichment coefficient of heavy metals such as As, Cu, Cr, and Pb in hook of U.rhynchophylla was being wasteland>woodland>cropland, the enrichment coefficient of Cu in hook of U. rhynchophylla in wasteland was more than 1. Except Cu, the enrichment coefficient of other heavy metals was low.

[Determination of Six Ingredients in Gardenia jaminoides Fruits with Quantitative Analysis of Muti-components by Single Marker].

OBJECTIVE: To establish a method of quantitative analysis of multi-components, by single marker(QAMS)for simultaneously determining six ingredients in Gardenia jasminoides fruits. METHODS: A multi-wavelength segmentation detection method was used. A methodological mode was found to analysis six ingredients in Gardenia jasminoides fruits by quantitative analysis of QAMS. Taken geniposide as reference to create RCF with gardenia acid, chlorogenic acid, crocin I, crocin II and crocin III. RESULTS: The good reproducibility and acceptable durability of method was validated between two HPLC systems and three columns. 20 batches of Gardenia jaminoides fruits was analysis, and the results showed good linear correlation compared to external standard method (r > 0. 999). CONCLUSION: QAMS can be used as quality evaluation method of multi-component Gardenia jaminoides fruits.

Intracellular Signaling Mechanisms Pharmacological Action of Jasminum amplexicaule Buch.-Ham. (Oleaceae) on Gastrointestinal Secretion.

Jasminum amplexicaule Buch-Ham. (Oleaceae) has been commonly used in the traditional medicine in dysentery, diarrhoea and bellyache in China. In the present work, the methanol extract of Jasminum amplexicaule (JME) was examined for pharmacology on human colonic epithelial cell line T84 by the short-circuit current technique. The results showed that pretreatment of T84 cells with JME produced a concentration-dependent (0-1000 µg/mL. EC50 = 0.055 mg/ mL) inhibition effect on adrenalin (Adr.)-induced Cl- secretion. The maximal response was observed at 200 µg/mL. It has been demonstrated that JME has a direct effect on the enterocyte. Our results also demonstrated that the JME exerted inhibitory effect on gastrointestinal Cl(-)secretion that effected by acting on basolateral ß-adrenoreceptors. These results suggested that the Chinese traditional medicine of JME can be used for the treatment of acute diarrhea and bellyache.

Preparative separation of alkaloids from Picrasma quassioides (D. Don) Benn. by conventional and pH-zone-refining countercurrent chromatography.

Two high-speed countercurrent chromatography (HSCCC) modes were compared by separation of major alkaloids from crude extract of Picrasma quassioides. The conventional HSCCC separation was performed with a two-phase solvent system composed of petroleum ether-ethyl acetate-methanol-water (5:5:4.5:5.5, v/v/v/v) with 200 mg loading. pH-Zone-refining CCC was performed with two-phase solvent system composed of petroleum ether-ethyl acetate-n-butanol-water (3:2:7:9, v/v/v/v) where triethylamine (10 mM) was added to the upper organic stationary phase and hydrochloric acid (5 mM) was added to the lower aqueous phase with 2 g loading. From 2 g of crude extract, 87 mg of 5-methoxycanthin-6-one (a), 38 mg of 1-methoxy-ß-carboline (b), 134 mg of 1-ethyl-4,8-dimethoxy-ß-carboline (c), 74 mg of 1-ethoxycarbonyl-ß-carboline (d), 56 mg of 1-vinyl-4,8-dimethoxy-ß-carboline (e) and 26 mg of 1-vinyl-4-dimethoxy-ß-carboline (f) were obtained with purities of over 97.0%. The results indicated that pH-zone-refining CCC is an excellent separations tool at the multigram level.

Preparative separation of six rhynchophylla alkaloids from Uncaria macrophylla wall by pH-zone refining counter-current chromatography.

pH-Zone refining counter-current chromatography was successfully applied to the preparative isolation and purification of six alkaloids from the ethanol extracts of Uncaria macrophylla Wall. Because of the low content of alkaloids (about 0.2%, w/w) in U. macrophylla Wall, the target compounds were enriched by pH-zone refining counter-current chromatography using a two-phase solvent system composed of petroleum ether-ethyl acetate-isopropanol-water (2:6:3:9, v/v), adding 10 mM triethylamine in organic stationary phase and 5 mM hydrochloric acid in aqueous mobile phase. Then pH-zone refining counter-current chromatography using the other two-phase solvent system was used for final purification. Six target compounds were finally isolated and purified by following two-phase solvent system composed of methyl tert-butyl ether (MTBE)-acetonitrile-water (4:0.5:5, v/v), adding triethylamine (TEA) (10 mM) to the organic phase and HCl (5 mM) to aqueous mobile phase. The separation of 2.8 g enriched total alkaloids yielded 36 mg hirsutine, 48 mg hirsuteine, 82 mg uncarine C, 73 mg uncarine E, 163 mg rhynchophylline, and 149 mg corynoxeine, all with purities above 96% as verified by HPLC Their structures were identified by electrospray ionization-mass spectrometry (ESI-MS) and 1H-NMR spectroscopy.

[Accumulation, distribution and pollution assessment of heavy metals in surface sediment of Caohai plateau wetland, Guizhou province].

The objective of this paper is to investigate the concentrations and distribution characteristics of heavy metals in surface sediments of different areas in the Caohai plateau wetland. 16 samples of surface sediments were collected and 7 heavy metals were analyzed. Heavy metal pollution in surface sediments of different areas in the Caohai plateau wetland was estimated by the Tomlinson Pollution Load Index (PLI) method. The analyzed results indicated that the average contents of Cd, Hg, As, Pb, Cr, Cu, Zn were 0.985, 0.345, 15.8, 38.9, 38.6, 22.8 and 384 mg x kg(-1), respectively. The heavy metal distributions varied with regional environment changes, the order of average contents of Cd and Hg in different regions was E (the eastern region) > S (the southern region) > N (the northern region), the order of the average content of Pb was N > E > S, and that of Zn was S > E > N. The results also suggested a medium heavy metal pollution level in the surface sediment of the Caohai plateau wetland with the PLI(zone) reaching 1.17. The order of pollution level in surface sediments of different regions was E > S > N. The results showed medium pollution levels in E and Hg which reached the extreme intensity pollution level were also the major polluted elements in surface sediments of the Caohai plateau wetland. And also, results showed medium pollution levels of Cd and Pb in surface sediments of Caohai plateau wetland. Cluster analysis results showed similar pollution sources of Cd, Zn, Pb and Hg, which should be attached great importance in terms of the prevention of the Caohai plateau wetland.

[Characteristic research on soil nutrient of root of Pseudostellaria heterophylla root zone and non-root zone in different planting patterns].

OBJECTIVE: To research the influence of Pseudostellaria heterophylla planting patterns on soil nutrients. METHODS: The root zone and non-root zone soil nutrient status were analyzed, which included the crop rotation, intercropping, relay cropping, 1-year planting, 3-year planting, 6-year planting and 10-year planting. RESULTS: The results showed that there were differences in the soil nutrient of root zone among different planting patterns. The soil nutrient of root zone in seven different planting patterns were all significantly higher than that of non-root zone soil. And the soil nutrients of root zone showed a significant enrichment. The enrichment ratio of total K and Olsen K in 6-year and 10-year planting were both negative value, which indicated that the soil potassium was deficiency. The average pH value of root zone was 0.32 lower than that of non-root zone. There was a significant positive correlation between the organic matter value and the total nitrogen content in root zone and non-root zone (P < 0.01). The soil nutrients were high in crop rotation and intercropping, it would he helpful to improve soil quality and absorption to nutrient. CONCLUSION: The reasonable application of nitrogen fertilizer ,organic fertilizer and potash fertilizer can significantly increase the organic matter content in soil ,and it will help to retain the fertilizer capacity of soil.

Qualitative and quantitative analyses of Epimedium wushanense by high-performance liquid chromatography coupled with diode array detection and electrospray ionization tandem mass spectrometry.

A new HPLC-DAD-ESI-MS(n) method was developed for rapid separation, characterization and quantitation of flavonoids in Epimedium wushanense, a popular Chinese herbal medicine. For qualitative identification, a total of 37 compounds were characterized from the underground and aerial parts of E. wushanense. Among them, 28 compounds were prenylated flavonoids, and 23 were confirmed by comparing with reference standards. For quantitative analysis, 12 major flavonoids including kaempferol glycosides, desmethylicaritin glycosides, and icaritin glycosides were simultaneously determined by HPLC/UV. Samples were separated on a Waters Symmetry C(18) column at 35 °C eluted with a gradient three-component mobile phase of acetonitrile, methanol, and water containing 0.03% v/v formic acid. All the flavonoids showed good linearity (r(2) ≥0.9997). The recoveries varied from 92.6 to 106.1% at three concentration levels. This method was applied to the determination of 20 samples of different geographical sources, harvesting time, and plant parts. Contents of the predominant flavonoid, epimedin C, ranged from 1.4 to 5.1% in aerial parts and 1.0 to 2.8% in underground parts. The methods established in this paper were simple and reliable and could be used for the quality control of E. wushanense.