Effects of Aminomethylphosphonic Acid on the Transcriptome and Metabolome of Red Swamp Crayfish, Procambarus clarkii.

Red swamp crayfish, Procambarus clarkii (P. clarkii), is an important model crustacean organism used in many types of research. However, the effects of different doses of aminomethylphosphonic acid (AMAP) on the transcriptome and metabolites of P. clarkii have not been explored. Thus, this study investigated the molecular and metabolic mechanisms activated at the different exposure dosages of AMAP in P. clarkii to provide new insights into the strategies of P. clarkii in response to the high concentrations of AMAP in the environment. In the present study, the P. clarkii were divided into three groups (control group; low-dosage AMAP exposure; high-dosage AMAP exposure), and hepatopancreatic tissue samples were dependently taken from the three groups. The response mechanisms at the different dosages of AMAP were investigated based on the transcriptome and metabolome data of P. clarkii. Differentially expressed genes and differentially abundant metabolites were identified in the distinct AMAP dosage exposure groups. The genes related to ribosome cell components were significantly up-regulated, suggesting that ribosomes play an essential role in responding to AMAP stress. The metabolite taurine, involved in the taurine and hypotaurine metabolism pathway, was significantly down-regulated. P. clarkii may provide feedback to counteract different dosages of AMAP via the upregulation of ribosome-related genes and multiple metabolic pathways. These key genes and metabolites play an important role in the response to AMAP stress to better prepare for survival in high AMAP concentrations.

Simultaneous removal of phosphate and tetracycline using LaFeO3 functionalised magnetic biochar by obtained ultrasound-assisted sol-gel pyrolysis: Mechanisms and characterisation.

Excessive phosphate and tetracycline (TC) contaminants pose a serious risk to human health and the ecological environment. As such exploring the simultaneous adsorption of phosphate and TC is garnering increasing attention. In this study, an efficient lanthanum ferrate magnetic biochar (FLBC) was synthesised from crab shells using an ultrasound-assisted sol-gel method to study its performance and mechanisms for phosphate and TC adsorption in aqueous solutions in mono/bis systems. According to the Langmuir model, the developed exhibited a maximum adsorption capacity of 65.62 mg/g for phosphate and 234.1 mg/g for TC (pH:7.0 ± 0.1, and 25 °C). Further, it exhibited high resistance to interference and pH suitability. In practical swine wastewater applications, whereby the concentrations of phosphate and TC are 37 and 19.97 mg/L, respectively, the proposed material demonstrated excellent performance. In addition, electrostatic adsorption, chemical precipitation and ligand exchange were noted to be the main mechanisms for phosphate adsorption by FLBC, whereas hydrogen bonding and π-π interaction were the main adsorption mechanisms for TC adsorption. Therefore, this study successfully prepared a novel and efficient adsorbent for phosphate and TC.

The Accumulation of Toxic Elements (Pb, Hg, Cd, As, and Cu) in Red Swamp Crayfish (Procambarus clarkii) in Qianjiang and the Associated Risks to Human Health.

Due to rapidly expanding crayfish consumption worldwide, the food safety of red swamp crayfish (Procambarus clarkii) is of great concern. China is the largest consumer and producer of crayfish globally. As of yet, it is unknown whether the main crayfish production cities in China are within safe levels of toxic heavy metals and metalloids. For 16 consecutive years, Qianjiang city ranked first in China in processing export volumes of red swamp crayfish. This study presents a comprehensive analysis of the enrichment levels and associated health risks of the species in Qianjiang. In our research, samples of four crayfish tissues, including the head, hepatopancreas, gills, and muscles, were collected from 38 sampling sites distributed in Qianjiang to evaluate the concentration levels of five heavy metals (Pb, Hg, Cd, As, and Cu). The concentration levels of all five metals in muscle did not surpass the national standard. Furthermore, eight significant correlations have been found. For further in-depth assess risk of crayfish in Qianjiang, estimated daily intake (EDI), target hazard quotient (THQ), carcinogenic risk (CR), and estimated maximum allowable consumption rates (CRmm) were evaluated in the abdomen muscle and hepatopancreas. The THQ values for each metal were found to be less than 1, while the CR values were below 10-6. Additionally, the CRmm for adults was determined to be 17.2 meals per month. These findings, based on the analysis of five metallic elements included in this study, suggest that the consumption of crayfish abdomen muscle in Qianjiang does not pose any significant health risks. However, it is noteworthy that certain regions exhibit elevated levels of arsenic in the hepatopancreas, surpassing the national standard, thereby rendering them unsuitable for excessive consumption. In general, the findings can be used to provide guidance for safe dietary practices in China.

Effect of flowing water on the pharmacokinetic properties of norfloxacin in channel catfish (Ictalurus punctatus) after single-dose oral administration.

BACKGROUND: The recirculating aquaculture system (RAS) is a widely used, water-saving and efficient aquaculture model. However, bacterial diseases are common in farmed fish reared at high densities. Although antibiotics effectively treat these diseases, developing efficient methods to increase drug clearance in fish and decrease the concentrations of antibiotic residues in aquatic products is essential. OBJECTIVES: This study evaluates the effect of flowing water in the RAS on norfloxacin (NOR) pharmacokinetics in channel catfish (Ictalurus punctatus). METHODS: Channel catfish were randomly divided into the control group (RAS group) and the experimental group (flow-through aquaculture system group) (120 individuals/group). A NOR dose of 20 mg/kg was then orally administered to the fish. Plasma, muscle, liver and kidney samples were collected up to 168 h after treatment. NOR concentrations were measured using liquid chromatography-mass spectrometry, and pharmacokinetic parameters were calculated using a non-compartmental method. RESULTS: Flowing water had a significant effect on the plasma pharmacokinetics and tissue distribution of NOR, increasing NOR clearance in the kidney, muscle and plasma. The time to maximum concentration of NOR was shorter in the plasma and longer in the kidney and liver. Moreover, flowing water increased the maximum concentration of NOR in the kidney, muscle and plasma and decreased the area under the concentration-time curve from time 0 to the last measurable concentration in the liver and plasma. Flowing water decreased the withdrawal period in muscle from 10 to 6 days. CONCLUSIONS: These results indicate that flowing water can potentially increase NOR clearance in channel catfish.

Oxidation towards enrofloxacin degradation over nanoscale zero-valent copper: mechanism and products.

Enrofloxacin (ENR) is a widely used veterinary fluoroquinolone antibiotic and is frequently detected in water environments. The degradation of ENR was examined utilizing molecular oxygen mediation using nanometer zero-valent copper (nZVC) as the catalyst in this work. The dosage of nZVC, initial pH, and reaction temperature were investigated as contributing factors to ENR degradation. The effects of Cl-, NO3-, SO42-, and humic acid on the degradation of ENR were investigated. The actual effects were evaluated using natural water. The reactive oxygen species (ROS) that participated in the reaction were identified, their generation mechanisms were elucidated, and the effects on ENR degradation were assessed. More emphasis was given to exploring ENR degradation and transformation pathways via analyses of HPLC-TOF-MS. Data showed that at 35 ℃, with an initial pH of 3 and exposed to air, an nZVC dose of 0.5 g·L-1 degraded ENR by 99.51% dramatically. HO• radicals were identified as the dominant ROS, and conversions among Cu0, Cu+, and Cu2+ played crucial roles in the generation of ROS. The destruction mechanism of ENR was speculated based on analyses of HPLC-TOF-MS results as the transformation of the piperazine ring into an oxidized state with a -COOH substitution with HO•, which caused ENR to be mineralized and converted into CO2, H2O, and [Formula: see text]. The ECOSAR program has been used to evaluate the toxicity of ENR and its degradation products, and oxidative degradation of nZVC significantly reduced its toxicity and increased its biodegradability. This research proposes a capable and practical method for removing ENR from water.

Mercury Induced Tissue Damage, Redox Metabolism, Ion Transport, Apoptosis, and Intestinal Microbiota Change in Red Swamp Crayfish (Procambarus clarkii): Application of Multi-Omics Analysis in Risk Assessment of Hg.

As one of the most toxic elements, mercury (Hg) is a widespread toxicant in aquatic environments. Crayfish are considered suitable for indicating the impact of heavy metals on aquatic crustaceans. Nevertheless, Hg toxicity on Procambarus clarkii is largely unknown. In this research, the acute Hg-induced alterations of biochemical responses, histopathology, hepatopancreatic transcriptome, and intestinal microbiome of Procambarus clarkii were studied. Firstly, Hg induced significant changes in reactive oxygen species (ROS) and malonaldehyde (MDA) content as well as antioxidant enzyme activity. Secondly, Hg exposure caused structural damage to the hepatopancreas (e.g., vacuolization of the epithelium and dilatation of the lumen) as well as to the intestines (e.g., dysregulation of lamina epithelialises and extension of lamina proprias). Thirdly, after treatment with three different concentrations of Hg, RNA-seq assays of the hepatopancreas revealed a large number of differentially expressed genes (DEGs) linked to a specific function. Among the DEGs, a lot of redox metabolism- (e.g., ACOX3, SMOX, GPX3, GLO1, and P4HA1), ion transport- (e.g., MICU3, MCTP, PYX, STEAP3, and SLC30A2), drug metabolism- (e.g., HSP70, HSP90A, CYP2L1, and CYP9E2), immune response- (e.g., SMAD4, HDAC1, and DUOX), and apoptosis-related genes (e.g., CTSL, CASP7, and BIRC2) were identified, which suggests that Hg exposure may perturb the redox equilibrium, disrupt the ion homeostasis, weaken immune response and ability, and cause apoptosis. Fourthly, bacterial 16S rRNA gene sequencing showed that Hg exposure decreased bacterial diversity and dysregulated intestinal microbiome composition. At the phylum level, there was a marked decrease in Proteobacteria and an increase in Firmicutes after exposure to high levels of Hg. With regards to genus, abundances of Bacteroides, Dysgonomonas, and Arcobacter were markedly dysregulated after Hg exposures. Our findings elucidate the mechanisms involved in Hg-mediated toxicity in aquatic crustaceans at the tissue, cellular, molecular as well as microbial levels.

Extracellular vesicles derived from human umbilical cord mesenchymal stem cells relieves diabetic retinopathy through a microRNA-30c-5p-dependent mechanism.

AIMS: Extracellular vesicle (EV)-transferred microRNAs (miRNAs) are proved to be potentially therapeutic candidates. Here, we attempted to unveil the role of delivery of miR-30c-5p by human umbilical cord mesenchymal stem cells (hUCMSCs)-derived EVs in diabetic retinopathy (DR). METHODS: miR-30c-5p and PLCG1 expression in streptozotocin-induced diabetes mellitus (DM) rats and high glucose (HG)-treated human retinal endothelial cells (HRECs) was quantified, followed by analysis on their interaction. EVs were isolated from hUCMSCs and co-cultured with HRECs. Through gain- and loss-of-function assays, the role of hUCMSCs-derived EV containing miR-30c-5p in DR involving PLCG1 and NF-κB pathway was analyzed in vitro and in vivo. RESULTS: Elevated PLCG1 was found in DM rats and HG-treated HRECs where miR-30c-5p was reduced while increased in hUCMSC-derived EVs. PLCG1 was pinpointed as a target gene of miR-30c-5p, which consequently disrupted the PKC/NF-κB pathway. hUCMSC-derived EVs decreased inflammation reaction by transferring miR-30c-5p in DM rats and HG-treated HRECs. Furthermore, similar changing tendency was observed in HG-treated HRECs induced by overexpressed miR-30c-5p through downregulation of PLCG1 in vivo. CONCLUSION: Overall, our findings underlined delivery of miR-30c-5p by hUCMSC-derived EVs as a novel suppressor in the inflammatory response following DR.

Effects of moderate exercise on hepatic amino acid and fatty acid composition, liver transcriptome, and intestinal microbiota in channel catfish (Ictalurus punctatus).

Previous research on swimming exercise in fish has focused on muscle building and quality of flesh. However, the effects of hepatic amino acid and fatty acid composition, liver gene expression profile, and intestinal microbiota are poorly understood. In this study, channel catfish (Ictalurus punctatus) were subjected to a 4-week swimming exercise, and liver transcriptome and intestinal microbiota analyses were performed to broaden our understanding of fish under exercise. When compared to non-exercised channel catfish (N-EXF), exercised channel catfish (EXF) had improved arachidonic acid (C20:4n6; ARA), docosahexaenoic acid (C22:6n3; DHA), aspartic acid (Asp) and glycine (Gly). The liver transcriptome analysis revealed 2912 differentially expressed genes and numerous enriched signaling pathways including those involved in nutrient synthesis, such as biosynthesis of unsaturated fatty acids and amino acids; glucose metabolism, such as glycolysis/gluconeogenesis, insulin signaling, and AMPK signaling pathways; and oxygen transport, such as HIF-1, PI3K-Akt, and MAPK signaling pathways. In addition, bacterial 16S rRNA gene sequencing data revealed that long-term exercise increased bacterial diversity and richness, and changed the intestinal microbial composition in channel catfish. In summary, this study provides insights into hepatic metabolic pathways, candidate genes, and intestinal microbiota underlying the long-term exercised channel catfish.

Comparative analyses of liver transcriptomes reveal the effect of exercise on growth-, glucose metabolism-, and oxygen transport-related genes and signaling pathways in grass carp (Ctenopharyngodon idella).

Grass carp is one of the most common farmed fish and its growth rate has been the focus of various studies. However, the impact of long-term exercise on growth rate of juvenile grass carp has not been clearly established. In this study, a four-month exercise trial and liver transcriptome analysis were performed to investigate changes in growth, liver molecular regulatory network and key genes in grass carp. When compared to the non-exercised grass carp (N-EXF), the exercised grass carp (EXF) showed a significant improvement in growth. Liver transcriptome analysis revealed 1714 significantly up-regulated and 1672 significantly down-regulated genes. These genes were enriched in various signaling pathways. These pathways included: those associated with growth, such as the PI3K-Akt and mTOR signaling pathways; those associated with glucose metabolism, such as glycolysis/gluconeogenesis, insulin and AMPK signaling pathways as well as those associated with oxygen transport, such as HIF-1, PI3K-Akt, PPAR and MAPK signaling pathways. In addition, growth-associated genes, such as ghr, igf1 and igf1r; glucose metabolism-associated genes, such as ins and insr as well as oxygen transport-associated genes, such as vhl, pdha and epo were identified. In conclusion, long-term moderate exercise improved the growth rate of grass carp. Our findings elucidate on changes in the liver molecular regulatory network and functional genes that occur during moderate exercise in fish.

Analysis of triazine herbicides in fish and seafood using a modified QuEChERS method followed by UHPLC-MS/MS.

A widely applicable method was established and validated for the qualitative and quantitative analysis of twenty-six triazine herbicides in multiple fish and seafood using ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). New convenient sample preparation approach based on modified QuEChERs was proposed by adding EMR-lipid to the traditional QuEChERS purification agents. Good separation was obtained after gradient elution through a C18 column. Target compounds were detected with electrospray ionization in positive ion mode and quantified in selective reaction monitoring (SRM) mode. Under this effective method, satisfactory results of linearity, sensitivity, accuracy, precision and matrix effect were achieved. Good linearities of the calibration curves yielded the correlation coefficients higher than 0.996 for all herbicides. The LOQs of 26 triazine herbicides ranged from 0.5 to 1.0 ng g-1. Good accuracy and precision were shown with the recoveries at the concentration of 1.0, 5.0 and 20.0 ng g-1 from 70.1% to 111.7% and the precisions of intra- and inter-day less than 12%. The matrix effects (-21.8-27.6%) were almost negligible. Finally the method was applied to the analysis of 72 crayfish samples from breeding bases of Hubei and Hunan provinces (China).

One step extraction followed by HPLC-ESI-MS/MS for multi-residue analysis of diacylhydrazine insecticides in water, sediment, and aquatic products.

A multi-residue analysis of six diacylhydrazine insecticides in water, sediment, and aquatic products was established by liquid chromatography triple quadrupole tandem mass spectrometry (LC-MS/MS). The water sample was extracted with acetonitrile by low-temperature enrichment liquid-liquid extraction technology. The sediment and aquatic products were prepared using QuEChERS technique. Method validation showed perfect linearity with correlation coefficients (R) more than 0.9992 for all insecticides, and the matrix effects were nearly negligible (-1.42% to -0.27%) for water, sediment and aquatic products. The recoveries were 80.0-99.7% at three spiked levels (0.02 ng·mL-1, 0.1 ng·mL-1, 0.5 ng·mL-1; 2.0, 10, and 50 ng·g-1) and the precisions (intra-day and inter-day precision) were lower than 5.28%, with the low LODs (3.8 ~ 9.6 pg·mL-1; 0.38-0.96 ng·g-1) and LOQs (12.7 ~ 32.0 pg·mL-1; 1.27-3.20 ng·g-1) for water, sediment, and aquatic products, indicating the good accuracy and precision of the proposed method. The applicability, efficiency, and sensitivity of this method have been proved in the analysis of six diacylhydrazine insecticides in water, sediment, and crucian carp in Rice- crucian carp - integrated planting system.

Determination of polychlorinated biphenyls and organochlorine pesticides in Chinese mitten crabs (Eriocheir sinensis) using modified QuEChERS followed by GC-MS.

Chinese mitten crabs (Eriocheir sinensis) are one of the most popular food items with a high nutritional value. The use of pesticides and environmental pollutants directly influenced improvements in productivity of mitten crabs. However, there is an increasing number of food safety issues raising consumer concerns. It is necessary to determine whether the pollutants that crabs contain are below the maximum residue limits to guarantee food safety. Moreover, there has been very little research on detecting pesticide residues and other pollutants in Chinese mitten crabs due to their complicated matrix effects. In the current study, a Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) method has been modified and validated for the simultaneous analysis of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) in mitten crabs. By using acetonitrile extraction, low temperature (freezing) precipitation and clean-up by mixed sorbents of C18, PSA and Florisil (magnesium silicate), the method was proved to be valid with linearity, precision, recovery, limit of quantification (LOQ), and limit of detection (LOD) with recoveries ranging from 85.9-119.8% and LOQ within 0.1-3.6 µg kg-1. The method was also applied to detect PCBs and OCPs in 48 crab samples from Jiangxi Province, China. The results showed that most of the crab samples contained PCB 118, beta-HCH and p,p'-DDT, and the content of p,p'-DDD in mitten crabs of Jiangxi Province showed sex-specific differences, which might be connected with sex-differential regulation.

High Glucose Induces the Loss of Retinal Pericytes Partly via NLRP3-Caspase-1-GSDMD-Mediated Pyroptosis.

Diabetic retinopathy (DR) is one of the hallmark complications of diabetes and a leading cause of vision loss in adults. Retinal pericyte death seems to be a prominent feature in the onset of DR. Pyroptosis is an inflammatory form of programmed cell death, defined as being caspase-gasdermin-D (GSDMD)-dependent. The NOD-like receptor pyrin 3 (NLRP3) inflammasome plays an important role in mediating GSDMD activation. However, the role and mechanism of pyroptosis in the loss of retinal pericytes during the pathogenesis of DR are still unclear. In the present study, we cultured primary human retinal pericytes (HRPs) in high glucose medium; caspase-3 inhibitor DEVD, caspase-1 inhibitor YVAD, or NLRP3 inhibitor glyburide was used as intervention reagents; GSDMD was overexpressed or suppressed by transfection with an expressing vector or retroviral silencing of GSDMD, respectively. Our data showed that high glucose induced NLRP3-caspase-1-GSDMD activation and pore formation in a dose- and time-dependent manner (p < 0.05) and resulted in the inflammatory cytokines IL-1ß and IL-18 and lactate dehydrogenase (LDH) release from HRPs (p < 0.05), which are all signs of HRP pyroptosis. Overexpression of GSDMD facilitated high glucose-induced pyroptosis (all p < 0.05). However, these effects were blunted by synergistically treating DEVD, YVAD, and silencing GSDMD (p < 0.05). Taken together, our results firstly revealed that high glucose induced the loss of retinal pericytes partly via NLRP3-caspase-1-GSDMD-mediated pyroptosis.

Molecular characterization of p38 MAPK and tissue-specific expression under cadmium stress in red swamp crayfish (Procambarus clarkii).

Keeping harmful pollutants out of some crucial tissues as much as possible is a key trait for the organism to survive in adverse conditions, such as in heavy-metal contaminated aquatic environments. In the current study, it was hypothesized that the p38 Mitogen-activated Protein Kinase (MAPK) controls the distribution of cadmium (Cd) in red swamp crayfish (Procambarus clarkii) by regulating the accumulation of Cd in different tissues under Cd-stressed conditions. To test this hypothesis, this study analyzed the p38 MAPK gene and compared the differential expression levels in the heart, antennal gland, gill, hepatopancreas, and muscles. Differences in expression levels of p38 MAPK gene between different tissues were conducted under controlled Cd exposure. This study found that the expression of p38 MAPK is tissue-specific in all tested samples under non-stressed condition. Under Cd-stressed condition and with the prolongation of Cd exposure time, the content of Cd in all examined tissues of P. clarkii has substantially increased compared to the control, although the Cd contents in the heart, antennal gland, and muscle remained relatively lower than those observed in the hepatopancreas. Consequently, the levels of p38 MAPK in the heart, antennal gland, and muscle were higher than the level in the hepatopancreas. These results indicate that p38 MAPK regulates the distribution and accumulation of Cd in different tissues of P. clarkii under Cd-stressed condition. Furthermore, the results suggested that the higher expression of p38 MAPK played a crucial role in keeping Cd out of the tissues in the Cd-rich aquatic environment to maintain the normal physiological function of the red swamp crayfish, a process necessary for survival and growth.

[Determination of carbendazim residues in Procambarus clarkii by high performance liquid chromatography-triple quadrupole mass spectrometry].

A novel method based on high performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-MS/MS) was established for the determination of carbendazim in Procambarus clarkii. The sample was extracted by ethyl acetate under alkaline conditions and centrifuged. The supernatant was concentrated by rotary evaporation, redissolved, and then enriched and purified on a mixed-mode cation exchange solid phase extraction column (MCX). A C18 column was used with acetonitrile and water as mobile phases in a gradient elution. Ionization was performed using an electrospray positive ion source (ESI+), and detection was achieved by MS/MS in multiple reaction monitoring (MRM) mode. The linearity for carbendazim was good in the range of 0.5-50.0 µg/L; the linear equation was y=0.19988x+0.01842; and the correlation coefficient (r2) was 0.9985. The limit of detection (LOD) and limit of quantification (LOQ) were 0.25 µg/kg and 0.50 µg/kg, respectively. At spiked levels of 0.5, 1.0, 5.0, and 50.0 µg/kg, the recoveries ranged from 83.9% to 105.5%, and the relative standard deviation ranged from 1.1% to 3.2%. This method is simple and effective for the determination of carbendazim residues in P. clarkii.

[Determination of chlorpyrifos residues in the paddy field aquatic products by ultra performance liquid chromatography-tandem mass spectrometry].

A method was developed for the determination of chlorpyrifos residues in paddy field aquatic products by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Samples were extracted with acetonitrile and purified by using 0.2 g primary secondary amine (PSA) sorbent and 1.2 g MgSO4. The target compound was separated on a Hypersil GOLD C18 column (100 mm×2.1 mm, 5 µm), and was determined using a heated electrospray ionization (HESI) source in the positive ion selective reaction monitoring (SRM) mode. The analyte was quantified with external standard using the matrix-matched standard calibration curve method. The results showed that good linearities were obtained in the range of 0.5-100.0 µg/L, and the correlation coefficients were greater than 0.999. The spiked recoveries of chlorpyrifos ranged from 86.2% to 103.6% with RSDs of 3.5%-7.6% (n=6). The limits of detection and quantification were 0.25 µg/kg and 0.5 µg/kg, respectively. This method is simple, quick, sensitive, and suitable for the rapid determination and analysis of chlorpyrifos residues in paddy field aquatic products.

Multi-residue method for the determination of organofluorine pesticides in fish tissue by liquid chromatography triple quadrupole tandem mass spectrometry.

A multiresidue method for the determination of organofluorine pesticides in fish tissue samples was developed and optimized. This method is based on a cleanup step of the extracted samples with acetonitrile, and carried out by solid phase extraction (SPE) with aminopropyl (NH2) cartridge, before the identification and quantification of the residues by liquid chromatography triple quadrupole tandem mass spectrometry (LC-MS/MS). The performance characteristics, such as accuracy, precision, linear range, limits of detection (LOD) and quantification (LOQ), for each pesticide were determined. The proposed method allowed high recoveries (80.4-99.2%) of spiked extracted fat samples at 0.5-10 ng g(-1), and very low LODs (between 0.10 and 0.15 ng g(-1)) and LOQs (between 0.3 and 0.5 ng g(-1)) determined in the fish samples. The practicality and high sensitivity of this method have been demonstrated in the determination of residues of OFPs in carp fish fed in water containing 10 ng mL(-1) OFPs.

c-Jun N-terminal kinase 3 expression in the retina of ocular hypertension mice: a possible target to reduce ganglion cell apoptosis.

Glaucoma, a type of optic neuropathy, is characterized by the loss of retinal ganglion cells. It remains controversial whether c-Jun N-terminal kinase (JNK) participates in the apoptosis of retinal ganglion cells in glaucoma. This study sought to explore a possible mechanism of action of JNK signaling pathway in glaucoma-induced retinal optic nerve damage. We established a mouse model of chronic ocular hypertension by reducing the aqueous humor followed by photocoagulation using the laser ignition method. Results showed significant pathological changes in the ocular tissues after the injury. Apoptosis of retinal ganglion cells increased with increased intraocular pressure, as did JNK3 mRNA expression in the retina. These data indicated that the increased expression of JNK3 mRNA was strongly associated with the increase in intraocular pressure in the retina, and correlated positively with the apoptosis of retinal ganglion cells.

Differential response to copper stress in the reproductive resources and allocation of metallophyte Kummerowia stipulacea.

Abundant seed production is a key life history trait for plant to maintain the stability of the whole population in adverse environments such as heavy metal contaminated mine area. In the current studies, we hypothesize that mine (metallicolous) populations of metallophytes have formed specialized reproductive strategies to adapt themselves to the heavy metal contaminated habitats, and differ from normal (non-metallicolous) populations in reproductive allocation. To test this hypothesis, the differences in reproductive resources and reproductive allocation between the copper mine and non-copper mine populations of pseudo-metallophyte Kummerowia stipulacea were comparatively examined under controlled Cu exposure experiments. Compared to non-copper mine population, copper mine population shows an increased seed output and larger reproductive effort under Cu stress. The increase of reproductive allocation in metallicolous population depends on not only seed size but also seed number per plant. The plants of metallicolous population increase allocation to the reproductive organs at the expense of a curtailment of allocation to vegetative traits, resulting in plants with shorter height and fewer branch numbers. There is little evidence displaying effect of root nodule on the reproductive resources and allocation. In addition, plants in metallicolous population reduce the transfer of Cu from roots to aboveground parts. These data suggest that plants of metallicolous population tend to invest more resources to reproductive output and increase their reproductive allocation in the adaptive evolution to Cu-enriched mine soils.

[Effect of electroacupuncture on the level of IL-1beta and TNF-alpha in patients with osteoarthritis].

OBJECTIVE: To study the therapeutic effect of electroacupuncture on patients with osteoarthritis, and to observe the effect of electroacupuncture on the level of IL-1beta and TNFalpha. METHODS: Forty-three subjects received electroacupuncture treatment at Liangqiu (ST 34), Xuehai (SP 10), Yinlingquan (SP 9), etc. The intensity of pain, the ability of the movement of legs, the level of IL-1beta and TNF-alpha in patients were measured before and after treatment. RESULTS: According to VAS, the intensity of pain was (64.22 +/- 14.55) and (32.50 +/- 10.79) before and after treatment, respectively. The score of movement ability of affected legs was (7.38 +/- 2.01) and (9.05 +/- 1.89) before and after treatment, respectively. Thus, the intensity of pain and the ability of movement were significantly improved after treatment (P < 0.01). The level of IL-1beta and TNF-alpha in patients were significantly decreased (both P < 0.05). CONCLUSION: The short term therapeutic effect of electroacupuncture on patients with osteoarthritis is better, it may result from that the electroacupuncture inhibit the activity of IL-1beta and TNF-alpha.