[Cloning and functional analysis of flavanone synthase â ¡ from Dendrobium huoshanense].

Flavonoid C-glycosides are a class of natural products that are widely involved in plant defense responses and have diverse pharmacological activities. They are also important active ingredients of Dendrobium huoshanense. Flavanone synthase Ⅱ has been proven to be a key enzyme in the synthesis pathway of flavonoid C-glycosides in plants, and their catalytic product 2-hydroxyflavanone is the precursor compound for the synthesis of various reported flavonoid C-glycosides. In this study, based on the reported amino acid sequence of flavanone synthase Ⅱ, a flavanone synthase Ⅱ gene(DhuFNSⅡ) was screened and verified from the constructed D. huoshanense genome localization database. Functional validation of the enzyme showed that it could in vitro catalyze naringenin and pinocembrin to produce apigenin and chrysin, respectively. The open reading frame(ORF) of DhuFNSⅡ was 1 644 bp in length, encoding 547 amino acids. Subcellular localization showed that the protein was localized on the endoplasmic reticulum. RT-qPCR results showed that DhuFNSⅡ had the highest expression in stems, followed by leaves and roots. The expression levels of DhuFNSⅡ and other target genes in various tissues of D. huoshanense were significantly up-regulated after four kinds of abiotic stresses commonly encountered in the growth process, but the extent of up-regulation varied among treatment groups, with drought and cold stress having more significant effects on gene expression levels. Through the identification and functional analysis of DhuFNSⅡ, this study is expected to contribute to the elucidation of the molecular mechanism of the formation of quality metabolites of D. huoshanense, flavonoid C-glycosides, and provide a reference for its quality formation and scientific cultivation.

Selenium alleviates biological toxicity of thiamethoxam (TMX): Bioaccumulation of TMX, organ damage, and antioxidant response of red swamp crayfish (Procambarus clarkii).

Pesticides are important for agricultural development; however, animals involved in rice-fish farming absorb the pesticides used during the farming process. Thiamethoxam (TMX) is extensively used in agriculture and is gradually occupying the market for traditional pesticides. Therefore, this study aimed to investigate whether selenomethionine (SeMet) could affect the survival rate, bioaccumulation of TMX, serum biochemical parameters, lipid peroxidation, antioxidants in the hepatopancreas, and expression of stress genes after exposure of red swamp crayfish to 10 ppt TMX for 7 days. The results showed that the survival rate significantly increased and the bioaccumulation of TMX significantly decreased with SeMet administration (P < 0.05). Furthermore, severe histological damage to the hepatopancreas of red crayfish was observed after exposure to TMX; however, this damage was alleviated after SeMet administration. SeMet also significantly reduced the TMX-induced changes in serum biochemical parameters, malondialdehyde content, and antioxidant enzyme activity in crayfish hepatopancreas (P < 0.05). Notably, analysis of the expression of 10 stress response genes showed that 0.5 mg/kg SeMet might decrease cell damage in the hepatopancreas. Consequently, our findings suggest that higher levels of TMX in crayfish may cause hepatopancreatic cell toxicity, which can be harmful to human health; however, SeMet could mitigate these effects, providing an understanding of pesticide compounds and food safety.

Bioaccumulation of Cd and comparative transcriptome analysis after the antagonism of Se in the hepatopancreas of estuary mud crab (Scylla paramamosain).

Cadmium (Cd) is a heavy metal contaminant and can be toxic to environment. What's more, Selenium (Se) protects organism as heavy metal antagonist. The present study aimed to investigate whether inorganic (Na2SeO3) or organic (L-SeMc) Se have an effect on the Cd bioaccumulation, antioxidant and immunity of the mud crab (Scylla paramamosain) under Cd exposure. The study showed that the concentration of Cd in hepatopancreas under Cd exposure was higher than the inorganic or organic Se group (P < 0.05), notably, Cd concentration of hepatopancreas in organic Se treatment is less than that in inorganic Se treatment (P < 0.05). Furthermore, this study analyzed 28 gene expression about antioxidant and immune from transcriptome, the result indicated that L-SeMc (organic Se) can reduced intracellular ROS production and oxidative damage. Furthermore, apoptosis was enhanced after Cd exposure, but Se could protect against apoptosis via expression of cathepsin B. Consequently, Organic Se may have a better effect than inorganic Se on reducing Cd toxicity. This study could provide the molecular basis that Se might alleviate Cd toxicity and increases the understanding of the environmental contaminant on crustaceans.

The health risk for consumers under heavy metal scenarios: Reduce bioaccumulation of Cd in estuary mud crab (Scylla paramamosain) through the antagonism of Se.

Heavy metal pollution has gained increasing attention over past years, and notably, cadmium (Cd) is a non-essential heavy metal that can be toxic to human and wildlife. Furthermore, selenium (Se) is a component of the selenoproteins and influences the toxicity of Cd in different organisms, and protect organisms as a kind of heavy metal antagonist. This study exposed mud crab to 5.0 mg/L Cd for 28 days, and investigated whether different concentrations (0.1, 0.2, 0.3 mg/kg) of selenite (Na2SeO3) or selenomethionine (SeMet) affect the bioaccumulation of Cd, serum biochemical index, antioxidant and stress-response genes of S. paramamosain. The results showed that the Cd concentration in Cd group was significantly higher than the organic or inorganic Se group. Serum biochemical index demonstrated that Se might relieve the damage or dysfunction of hepatopancreas caused by both Cd accumulation and toxicity. Furthermore, Se improved CAT, GPx T-AOC and SOD activity, and decreased MDA concentrations and the lipid peroxidation levels, antagonistic to Cd. Then, this study analyzed the expression of 26 stress-related genes, the results indicated that the inorganic and organic Se might reduce the damage of cell and the toxicity of heavy metals in the hepatopancreas after Cd exposure. Therefore, this study indicated that Se might alleviate Cd toxicity via the different antioxidative mechanisms, and increased the understanding of environmental toxins on estuary crustaceans.

[Acupoint thread-embedding as adjuvant therapy for bronchial asthma in children and its effect on pulmonary function, serum IgA and IgE levels].

OBJECTIVE: To observe the efficacy of acupoint thread-embedding combined with fluticasone propionate aerosol for chronic persistent bronchial asthma of lung-spleen qi deficiency and spleen-kidney yang deficiency in children and its effect on pulmonary function and serum levels of IgA and IgE. METHODS: A total of 120 children with chronic persistent bronchial asthma were randomly divided into an observation group (60 cases, 9 cases dropped off) and a control group (60 cases, 7 cases dropped off). The control group was treated with fluticasone propionate aerosol (125 µg per inhalation), twice a day; based on the control group treatment, the observation group was treated with acupoint thread-embedding at Dingchuan (EX-B 1), Feishu (BL 13), Zusanli (ST 36) and Danzhong (CV 17), once half a month. Both groups were treated for 3 months. The pulmonary function, serum IgA, IgE levels and TCM symptom score were compared between the two groups before and after treatment, and the clinical efficacy was evaluated. RESULTS: After treatment, the large airway function (peak expiratory flow [PEF], forced expiratory volume at the first second [FEV1]) and small airway function (maximal expiratory flow at 25% of the forced capacity [MEF25%], maximal expiratory flow at 50% of the forced capacity [MEF50%], maximal expiratory flow at 75% of the forced capacity [MEF75%] and midexpiratory flow 25%-75% [MEF25%-75%]) were higher than those before treatment (P<0.05), and the pulmonary function in the observation group was higher than that in the control group (P<0.05). After treatment, the serum IgA levels in the two groups were higher than those before treatment (P<0.05), and the serum IgE levels and TCM symptom scores were lower than those before treatment (P<0.05); the serum IgA level in the observation group was higher than that in the control group (P<0.05), and the serum IgE level and TCM symptom score in the observation group were lower than those in the control group (P<0.05). The total effective rate was 94.1% (48/51) in the observation group, which was higher than 88.7% (47/53) in the control group (P<0.05). CONCLUSION: Acupoint thread-embedding combined with fluticasone propionate aerosol could improve the pulmonary function, TCM symptoms and serum IgA and IgE levels in children with chronic persistent bronchial asthma of lung-spleen qi deficiency and spleen-kidney yang deficiency. The curative effect is better than fluticasone propionate aerosol alone.

Dietary values of macroalgae Porphyra haitanensis in Litopenaeus vannamei under normal rearing and WSSV challenge conditions: Effect on growth, immune response and intestinal microbiota.

Two trials were conducted to determine the effects of dietary macroalgae Porphyra haitanensis on growth, immunity and intestinal microbiota of Litopenaeus vannamei. In trial 1, shrimp (mean initial wet weight about 0.64 g) were fed with seven diets (P0, P1, P2, P3, P4, P5 and P6) containing 0% (basal diet), 1%, 2%, 3%, 4%, 5% and 6% P. haitanensis in triplicate for 60 days. Growth performance (weight gain, WG; specific growth rate, SGR) of shrimp fed the P4 diet were significantly higher than that of shrimp fed P0, P5 and P6 diets (P < 0.05) but without significant differences with shrimp fed P1-P3 diets (P > 0.05). Hepatopancreas phenoloxidase (PO) activity of shrimp fed the P. haitanensis containing diets was significantly higher than that of shrimp fed the basal diet (P0) (P < 0.05). Total haemocyte count (THC) of shrimp fed basal diet (P0) was significantly lower than that of shrimp fed diets containing P. haitanensis. Our results declared that dietary P. haitanensis supplementation increases the abundance of beneficial bacterials such as Nitrosopumilus, Marinobacter or Bifidobacterium and reduces the abundance of harmful bacterias such as Vibrio, and especially pronounced in P4 diet treatment. In trial 2, a WSSV injection challenge test was conducted for 7-day after the rearing trial and shrimp survival was also compared among treatments. A sudden shrimp death was found from the 4th day, and values of survival of shrimp fed the P3-P4 diets were higher than that of shrimp fed other diets during 4-7 days challenge test. The immune response in trial 2 were characterized by higher superoxide dismutase activity (SOD) and PO activities, lower THC and higher HCT compared to levels found in trial 1. In conclusion, suitable dietary P. haitanensis could enhance the growth performance, antioxidant capacity and alter total bacterial numbers or microbial diversity of L. vannamei and furthermore reduce oxidative stress and immune depression challenged by WSSV injection stress, and the level of P. haitanensis supplemented in the diet should be between 2.51% and 3.14%.

Dietary values of Forsythia suspensa extract in Penaeus monodon under normal rearing and Vibrio parahaemolyticus 3HP (VP3HP) challenge conditions: Effect on growth, intestinal barrier function, immune response and immune related gene expression.

Two trials were conducted to determine the effects of dietary Forsythia suspensa extract (FSE) on shrimp, Penaeus monodon, first on growth performance, second on the immune response and immune related gene expression of shrimp. In trial 1, shrimp (mean initial wet weight about 3.02 g) were fed with five diets containing 0% (basal diet), 0.01%, 0.02%, 0.04% and 0.06% FSE in triplicate for 60 days. Growth performance (final body wet weight, FBW; weight gain, WG; biomass gain, BG) of shrimp fed FSE diets were higher (P < 0.05) than that of shrimp fed the basal diet. The survival among all the diets treatments were above 90% and no significant difference was revealed among them (P > 0.05). The antioxidant capacity (total antioxidant status, TAS; glutathione peroxidase, GSH-Px) appears in the trend of firstly increasing then decreasing with the increasing of dietary FSE levels. The highest value of TAS and GSH-Px were found in shrimp fed 0.02% FSE diet and were significantly higher than that of shrimp fed the basal and 0.06% FSE diets (P < 0.05). Hepatopancreas malondialdehyde (MDA) of shrimp fed FSE diets were lower (P < 0.05) than that of shrimp fed the basal diet. Total haemocyte count of shrimp fed the basal diet was lower (P < 0.05) than that of shrimp fed FSE diets. Haemolymph clotting time of shrimp had the opposite trend with the total haemocyte count of shrimp. No significant differences were found in haemolymph biomarkers of intestinal permeability (endotoxin and diamine oxidase) and in molecular gene expression profiles of heat shock protein 70 (Hsp 70) mRNA and hypoxia inducible factor-1α (HIF-1α) mRNA in haemolymph of shrimp among all diet treatments (P > 0.05). In trial 2, a pathogenic strain of Vibrio parahaemolyticus 3HP (VP3HP) injection challenge test was conducted for 6-day after the rearing trial and shrimp survival were also compared among treatments. Survival of shrimp fed diets supplemented with 0.01%-0.02% FSE were higher than that of shrimp fed the basal and 0.06% FSE diets (P < 0.05). Dietary FSE supplementation produced stronger hepatopancreas antioxidant capacity (TAS, GSH-Px) (P < 0.05) and higher glutathione (GSH) level (P < 0.05), lower superoxide dismutase activity (SOD) (P < 0.05), higher total haemocyte count (P < 0.05), lower haemolymph clotting time (P < 0.05), lower MDA and carbonyl protein concentration (P < 0.05), lower haemolymph biomarkers of intestinal permeability (endotoxin and diamine oxidase) (P < 0.05), generated lower molecular gene expression profiles of HSP 70 mRNA and higher HIF-1α mRNA (P < 0.05) than the basal diet. The immune response were characterized by lower TAS and higher antioxidant enzyme activities (SOD, GSH-Px) and higher oxidative stress level (MDA and carbonyl protein) and higher haemolymph biomarkers of intestinal permeability (endotoxin and diamine oxidase) compared to levels found in trail 1. However, the total haemocyte counts and haemolymph clotting times were not changed in 0.01%-0.02% FSE diets treatments between trial 1 and trial 2 (P > 0.05). The molecular gene expression profile of Hsp 70 mRNA was increased while HIF-1α mRNA was decreased when compared to trial 1. In conclusion, results suggested that dietary intake containing FSE could enhance the growth performance and antioxidant capacity of P. monodon and furthermore reduce oxidative stress and immune depression challenged by a pathogenic strain of Vibrio parahaemolyticus stress. Considering the effect of FSE on both growth performance and immune response of P. monodon, the level of FSE supplemented in the diet should be between 0.01% and 0.02%.

Cardiac electrophysiological and antiarrhythmic effects of N-n-butyl haloperidol iodide.

AIMS: N-n-butyl haloperidol (F(2)), a novel compound of quaternary ammonium salt derivatives of haloperidol, was reported to antagonize myocardial ischemia/reperfusion injuries. The antiarrhythmic potential and electrophysiological effects of F(2) on rat cardiac tissues were investigated. METHODS AND RESULTS: In Langendorff-perfused rat hearts, the ventricular arrhythmias were induced by left anterior descending coronary artery of rat heart ligated for 20 min before the release of the ligature. F(2) provided some inhibitive effects against ischemia- and reperfusion-induced ventricular arrhythmias. In His bundle electrogram and epicardial ECG recordings, the drug produced bradycardia, delayed the conduction through the atrioventricular node and prolonged the Wenckebach cycle length and atrioventricular nodal effective refractory period. In whole-cell patch-clamp study, F(2) primarily inhibited the L-type Ca2+ current (I(Ca,L)) (IC(50) = 0.17 microM) with tonic blocking properties and little use-dependence. And the drug also decreased the Na+ current (IC(50) = 77.5 microM), the transient outward K+ current (IC(50) = 20.4 microM), the steady-state outward K+ current (IC(50) = 56.2 microM) and the inward rectifier K+ current (IC(50) = 127.3 microM). CONCLUSION: F(2) may be a promising drug for the treatment of ischemic heart disease with cardiac arrhythmia.