Horizontal and Vertical Transmission of a Mycovirus Closely Related to the Partitivirus RhsV717 That Confers Hypovirulence in Rhizoctonia solani.

Rhizoctonia solani virus717 (RhsV717) was isolated from the Rhizoctonia solani (R. solani) AG-2 strain Rhs717. This study isolated a virus designated as Rhizoctonia solani partitivirus BS-5 (RsPV-BS5) from the R. solani AG-3 strain BS-5, the causal agent of tobacco target spot disease. The virus was identified as a strain of RhsV717. Transmission electron microscopy (TEM) images showed that RsPV-BS5 had virus particles with a diameter of approximately 40 nm. Importantly, it can be horizontally transmitted through hyphal anastomosis and vertically transmitted via sexual basidiospores. Furthermore, this study demonstrated that RsPV-BS5 infection significantly impedes mycelial growth and induces hypovirulence in tobacco leaves. Thus, RsPV-BS5 presents a promising avenue for biocontrolling tobacco target spot disease. Transcriptome analysis unveiled differential expression of four genes related to cell wall-degrading enzymes between two isogenic strains, 06-2-15V and 06-2-15. These findings shed light on the molecular mechanism through which RsPV-BS5 reduces host pathogenicity.

Complete genome sequence of a novel fusarivirus from Rhizoctonia solani AG-3 PT strain 3P-2-2.

Here, we describe a novel mycovirus, tentatively designated as "Rhizoctonia solani fusarivirus 6" (RsFV6), which was discovered in Rhizoctonia solani AG-3 PT strain 3P-2-2. The virus has a single-stranded positive-sense RNA (+ssRNA) genome of 6141 nucleotides containing two open reading frames (ORFs) and a poly(A) tail. ORF1 encodes a large polypeptide of 1,862 amino acids (aa) with conserved RNA-dependent RNA polymerase (RdRp) and helicase (Hel) domains. ORF2 encodes a putative 167-aa protein of unknown function. BLASTp searches revealed that the ORF1-encoded polypeptide showed the highest sequence similarity (70.67% identity) to that of Rhizoctonia solani fusarivirus 3 (RsFV3), which was isolated from Rhizoctonia solani AG-2-2LP. Multiple sequence alignments and phylogenetic analysis based on RdRp and Hel sequences indicated that RsFV6 could be a novel member of the genus Alphafusarivirus family Fusariviridae.

Occurrence, source, and ecological risk assessment of organochlorine pesticides and polychlorinated biphenyls in the water-sediment system of Hangzhou Bay and East China Sea.

The pollution characteristics, potential sources, and potential ecological risk of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) were investigated in the Hangzhou Bay (HZB) and East China Sea (ECS). Total OCPs concentration ranged from 2.62 to 102.07 ng/L and 4.41 to 75.79 µg/kg in the seawater and sediment samples, with PCBs concentration in the range of 0.40-51.75 ng/L and 0.80-45.54 µg/kg, respectively. The OCPs were positively correlated with nutrients, whereas PCBs presented a negative correlation. The newly imported dichlorodiphenyltrichloroethane (DDT) in HZB is mainly the mixing of technical DDT and dicofol sources. The PCB source composition is more likely related to the mixture of Kanechlor 300, 400, Aroclor 1016, 1242, and Aroclor 1248. Risk assessment results indicate that OCPs posed low risk in seawater. The potential risk of DDTs in the sediments is a cause of concern.

Spatiotemporal occurrence, sources and risk assessment of polycyclic aromatic hydrocarbons in a typical mariculture ecosystem.

The spatiotemporal variations, influencing factors and potential sources, as well as the ecological/health risks of polycyclic aromatic hydrocarbons (PAHs) were systematically investigated in seawater, sediment, and fish from Xiangshan Bay, China, one of the most important and oldest domestic marine aquaculture bases. The average concentrations of ΣPAHs in seawater, sediment and fish were 150 ± 70.0 ng/L, 276 ± 271 µg/kg (dry weight, dw), and 434 ± 151 µg/kg (dw), respectively. Naphthalene, phenanthrene, fluoranthene, benzo(b)fluoranthene and pyrene were the dominant contaminants in all samples. The highest PAH concentrations in the seawater and sediment samples occurred in the inner bay where the mariculture and industry are clustered. Seasonal differences were observed in the seawater samples but not in the sediment samples. Among all 15 fish species, large yellow croaker (Larimichthys crocea) (775 µg/kg (dw)), red drum (Sciaenops ocellatus) (749 µg/kg (dw)), and flathead grey mullet (Mugil cephalus) (637 µg/kg (dw)) had relatively high PAH accumulation concentrations in muscle tissue. According to the molecular diagnostic ratio method, the PAHs in seawater mainly originated from a mixed source of petroleum and combustion, whereas biomass/coal combustion sources were identified for sediment. The results obtained from the risk quotient (for seawater), sediment quality guidelines and toxic equivalence quotients (for seawater and sediment) methods showed that the ecological risks posed by PAHs were generally at a low to moderate level. Potentially toxic effects existed from PAH-contaminated fish consumption, and the resulting potential carcinogenic risk was also slightly higher than the recommended guidelines (10-6).

Probing the contamination characteristics, mobility, and risk assessments of typical plastic additive-phthalate esters from a typical coastal aquaculture area, China.

Contamination characteristics, equilibrium partitioning and risk assessment of phthalate esters (PAEs) were investigated in seawater, sediment and biological samples collected from the Xiangshan Bay area during an annual investigation between January and November 2019. PAE concentrations detected in the mariculture environment in surface seawater, sediment, and biological samples were 172-3365 ng/L, 190-2430 µg/kg (dry weight [dw]), and 820-4926 µg/kg (dw), respectively. The dominant congeners in different media included di-n-butyl phthalate (DnBP), diisobutyl phthalate (DiBP), and di(2-ethylhexyl) phthalate (DEHP). The inner bay and the bay mouth were the gathering area of PAEs and heavily influenced by the mariculture activities, river inputs, and anthropogenic activities. The bioaccumulation of PAEs demonstrated benthic feeding fishes with relatively high trophic levels concentrated high levels of phthalates. The mobility of PAEs in sediment-seawater showed that the transfer tendency of low-molecular weight species was from the sediment to the water, which was in contrast with those of high-molecular weight PAEs. DEHP, DiBP and DnBP had various degrees of ecological risks in the aquatic environment, whereas only the DiBP posed potential risks in sediments. The current assessment of carcinogenic and noncarcinogenic risks posed by fish consumption were within acceptable limits for humans.

Platinum nanozyme-encapsulated poly(amidoamine) dendrimer for voltammetric immunoassay of pro-gastrin-releasing peptide.

An innovative electrochemical immunosensing platform was designed for the sensitive monitoring of lung cancer biomarker (pro-gastrin-releasing peptide; ProGRP) by using platinum nanoparticles encapsulated inside dendrimers (PtDEN) as enzymatic mimics for the signal amplification. PtDEN nanocomposites were prepared through a simple chemical reduction method with the assistance of NaBH4. Thereafter, PtDEN-labeled anti-ProGRP secondary antibody was launched for the detection of target analyte with a sandwich-type assay format on anti-ProGRP capture antibody-modified screen-printed carbon electrode. Accompanying formation of immunocomplex, the labeled PtDENs electrochemically oxidized 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of hydrogen peroxide to produce a well-defined voltammetric signal within the applied potentials. Thanks to the high-efficient catalytic efficiency of platinum nanoparticles and high-loading ability of dendrimer, improved analytical features were acquired with PtDENs relative to platinum nanoparticles alone. Using PtDENs labeling strategy, the properties and factors influencing the analytical performance of electrochemical immunosensor were studied in detail. The strong bioconjugation of antibodies with the PtDENs caused a good repeatability and intermediate precision down to 7.64%. Under optimum conditions, the electrochemical immunosensor exhibited a dynamic linear range of 0.001-10 ng mL-1 ProGRP with a detection limit of 0.86 pg mL-1. Good selectivity and relatively long-term stability (>6 months) were achieved for target ProGRP. Significantly, the acceptable accuracy was gotten for analysis of ProGRP in human serum specimens referring to commercially available human ProGRP enzyme-linked immunosorbent assay (ELISA) method.

Lipin1 Is Involved in the Pathogenesis of Diabetic Encephalopathy through the PKD/Limk/Cofilin Signaling Pathway.

Diabetic encephalopathy is a type of central diabetic neuropathy resulting from diabetes mainly manifested as cognitive impairments. However, its underlying pathogenesis and effective treatment strategies remain unclear. In the present study, we investigated the effect of Lipin1, a phosphatidic acid phosphatase enzyme, on the pathogenesis of diabetic encephalopathy. We found that in vitro, Lipin1 exerts protective effects on high glucose-induced reductions of PC12 cell viability, while in vivo, Lipin1 is downregulated within the CA1 hippocampal region in a type I diabetes rat model. Increased levels of Lipin1 within the CA1 region are accompanied with protective effects including amelioration of dendritic spine and synaptic deficiencies, phosphorylation of the synaptic plasticity-related proteins, LIM kinase 1 (p-limk1) and cofilin, as well as increases in the synthesis of diacylglycerol (DAG), and the expression of phosphorylated protein kinase D (p-PKD). These effects are associated with the rescue of cognitive disorders as shown in this rat model of diabetes. In contrast, knockdown of Lipin1 within the CA1 region enhanced neuronal abnormalities and the genesis of cognitive impairment in rats. These results suggest that Lipin1 may exert neuroprotective effects involving the PKD/Limk/Cofilin signaling pathway and may serve as a potential therapeutic target for diabetic encephalopathy.

Cross-linkage urease nanoparticles: a high-efficiency signal-generation tag for portable pH meter-based electrochemical immunoassay of lipocalin-2 protein diagnostics.

An innovative signal-transduction tag based on cross-linked urease nanoparticles (CLENP) was designed for the development of a pH meter-based immunoassay of lipocalin-2 (LCN2). The CLENP was synthesized with a typical desolvation method using ethanol as desolvation agent, followed by functionalization with polyaspartic acid. The carboxylated CLENP were used as the signal-generation tags for the labelling of secondary antibodies via the carbodiimide coupling. Upon target LCN2 introduction, a sandwich-type immune reaction was performed between capture antibody-coated plate and the labeled secondary antibody on the CLENP. The conjugated CLENP in the microplate hydrolyzed urea into ammonia (NH4+) and carbonate (CO32-), resulting in the pH change of solution, which was determined with a handheld pH meter. The pH variation was proportional to target concentration in the sample. By monitoring the pH variation of the urea solution, the level of LCN2 at a concentration as low as 5.2 pg mL-1 was evaluated. The pH meter-based electrochemical immunoassay can be utilized for mass production of miniaturized lab-on-a-chip devices with handheld pH meter, thereby opening new opportunities for protein diagnostics and biosecurity. Graphical abstract An innovative signal-transduction tag based on cross-linked urease nanoparticles was designed for high-efficiency immunoassay of lipocalin-2 with pH meter readout.

Effective removal matrix interferences by a modified QuEChERS based on the molecularly imprinted polymers for determination of 84 polychlorinated biphenyls and organochlorine pesticides in shellfish samples.

A modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) procedure combined with GC-MS/MS detection approach using a dynamic multiple reaction monitoring (DMRM) mode was successfully applied for the simultaneous analysis of 84 polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) in shellfish samples. The novel molecular imprinted polymers (MIPs) were synthesized by precipitation polymerization and characterized by Scanning electron microscopy, Brunauer-Emmett-Teller, Fourier transform infrared spectra and adsorption experiment. The MIPs exhibited good adsorption capability to pigment coextractives in shellfish samples without the loss of analytes compared with other sorbents. Under optimal conditions, spiked experiments in sinonovacula, mussel, and clam at 10.0-100.0 µg/kg concentrations showed excellent recoveries ranging from 70% to 120% for all analytes with the relative standard deviations of <10%. The developed method showed good linearity with the correlation coefficient above 0.9980, and the limits of quantification were in the range of 0.01-9.02 µg/kg. The developed QuEChERS procedure combined with GC-MS/MS was successfully applied to 84 PCBs and OCPs residues detection in shellfish samples.

Silver-functionalized g-C3N4 nanohybrids as signal-transduction tags for electrochemical immunoassay of human carbohydrate antigen 19-9.

A simple and feasible electrochemical immunosensing platform was developed for highly efficient screening of a disease-related protein (human carbohydrate antigen 19-9, CA 19-9 used in this case) using silver-functionalized g-C3N4 nanosheets (Ag/g-C3N4) as signal-transduction tags. Initially, Ag/g-C3N4 nanohybrids were synthesized by combining thermal polymerization of the melamine precursor with the photo-assisted reduction method. Thereafter, the as-synthesized Ag/g-C3N4 nanohybrids were utilized for the labeling of the anti-CA 19-9 detection antibody by using a typical carbodiimide coupling method. The assay was carried out on a capture antibody-modified glassy carbon electrode in a sandwich-type detection mode. The detectable signal mainly derived from the voltammetric characteristics of the immobilized nanosilver particles on the g-C3N4 nanosheets within the applied potentials. Under the optimal conditions, the voltammetric peak currents increased with the increasing amount of target CA 19-9, and exhibited a wide linear range from 5.0 mU mL(-1) to 50 U mL(-1) with a detection limit of 1.2 mU mL(-1). Our strategy also displayed good reproducibility, precision and specificity. The results of the analysis of clinical serum specimens were in good accordance with the results obtained by an enzyme-linked immunosorbent assay (ELISA) method. The newly developed immunosensing system is promising for enzyme-free and cost-effective analysis of low-abundance proteins.

The role of long-term label-retaining cells in the regeneration of adult mouse kidney after ischemia/reperfusion injury.

BACKGROUND: Label-retaining cells (LRCs) have been recognized as rare stem and progenitor-like cells, but their complex biological features in renal repair at the cellular level have never been reported. This study was conducted to evaluate whether LRCs in kidney are indeed renal stem/progenitor cells and to delineate their potential role in kidney regeneration. METHODS: We utilized a long-term pulse chase of 5-bromo-2'-deoxyuridine (BrdU)-labeled cells in C57BL/6J mice to identify renal LRCs. We tracked the precise morphological characteristics and locations of BrdU(+)LRCs by both immunohistochemistry and immunofluorescence. To examine whether these BrdU(+)LRCs contribute to the repair of acute kidney injury, we analyzed biological characteristics of BrdU(+)LRCs in mice after ischemia/reperfusion (I/R) injury. RESULTS: The findings revealed that the nuclei of BrdU(+) LRCs exhibited different morphological characteristics in normal adult kidneys, including nuclei in pairs or scattered, fragmented or intact, strongly or weakly positive. Only 24.3 ± 1.5 % of BrdU(+) LRCs co-expressed with Ki67 and 9.1 ± 1.4 % of BrdU(+) LRCs were positive for TUNEL following renal I/R injury. Interestingly, we found that newly regenerated cells formed a niche-like structure and LRCs in pairs tended to locate in this structure, but the number of those LRCs was very low. We found a few scattered LRCs co-expressed Lotus tetragonolobus agglutinin (LTA) in the early phase of injury, suggesting differentiation of those LRCs in mouse kidney. CONCLUSIONS: Our findings suggest that LRCs are not a simple type of slow-cycling cells in adult kidneys, indicating a limited role of these cells in the regeneration of I/R injured kidney. Thus, LRCs cannot reliably be considered stem/progenitor cells in the regeneration of adult mouse kidney. When researchers use this technique to study the cellular basis of renal repair, these complex features of renal LRCs and the purity of real stem cells among renal LRCs should be considered.

Sensitive electrochemical immunoassay with signal enhancement based on nanogold-encapsulated poly(amidoamine) dendrimer-stimulated hydrogen evolution reaction.

A new electrochemical immunosensor with signal enhancement was designed for sensitive detection of disease-related protein (human carbohydrate antigen 19-9, CA 19-9 used in this case). The assay was carried out on a capture antibody-modified screen-printed carbon electrode with a sandwich-type mode by using detection antibody-functionalized nanogold-encapsulated poly(amidoamine) dendrimer (AuNP-PAAD). The AuNP-PAAD was first synthesized through the in situ reduction method and functionalized with the polyclonal rabbit anti-human CA 19-9 antibody. Upon target CA 19-9 introduction, a sandwiched immunocomplex could be formed between the capture antibody and detection antibody. Accompanying the AuNP-PAAD, the electrocatalytic activity of the carried gold nanoparticles toward the hydrogen evolution reaction (HER) allowed the rapid quantification of the target analyte on the electrode. The amplified electrochemical signal mainly derived from AuNP-catalyzed HER in an acidic medium. Under optimal conditions, the immuno-HER assay displayed a wide dynamic concentration range from 0.01 to 300 U mL(-1) toward target CA 19-9 with a detection limit (LOD) of 6.3 mU mL(-1). The reproducibility, precision, specificity and stability of our strategy were acceptable. Additionally, the system was further validated by assaying 13 human serum specimens, giving well matched results obtained from the commercialized enzyme-linked immunosorbent assay (ELISA) method.

Vaspin as a Risk Factor of Insulin Resistance in Obstructive Sleep Apnea-Hypopnea Syndrome in an Animal Model.

BACKGROUND: In this study, we aimed to establish a chronic intermittent hypoxia model in rats and explore the possible role of vaspin in insulin sensitivity. METHODS: Healthy male Wistar rats were randomly divided into two groups: normal control group (NC) and chronic intermittent hypoxia group (CIH). The NC group was raised under physiological conditions and the CIH group was kept in the plexiglass chamber between 9 am and 5 pm undergoing intermittent hypoxic challenge for 8 hours/day for 8 weeks. Arterial blood pressure of rats (tail cannulation) was measured before and after the study. Fasting plasma glucose (FPG), total cholesterol (TC), triglycerides (TG), fasting insulin (FINS), vaspin, and leptin levels were measured. Vaspin mRNA expression in visceral adipose tissues was measured with Real Time-PCR. The protein levels of vaspin, Akt and phospho-Akt in visceral tissues were determined by Western-blot. RESULTS: At baseline, all the measurements in the CIH and NC groups were comparable. By the end of the experiment, the blood pressure of the CIH group was significantly higher than the NC group. The levels of FPG, FINS, TG, TC, leptin, and vaspin in the CIH group were significantly higher than in NC group. Plasma vaspin levels were correlated with FINS, HOMA-IR, and TG levels. Vaspin expression in both mRNA and protein levels in visceral adipose tissues of the CIH group were clearly higher than the NC group. Phospho-Akt protein level was decreased in visceral adipose tissues of the CIH group compared to the NC group. CONCLUSIONS: In the chronic intermittent hypoxia rat model, the expression of vaspin in visceral adipose tissues and plasma were increased, which were correlated with insulin resistance.

Exonuclease III-based target recycling for ultrasensitive homogeneous monitoring of HIV DNA using Ag(+)-coordinated hairpin probe.

A new homogeneous electrochemical sensing strategy based on exonuclease III-assisted target recycling amplification was utilized for simple, rapid and highly sensitive detection of human immunodeficiency virus (HIV) DNA on an immobilization-free Ag(I)-assisted hairpin DNA through the cytosine-Ag(+)-cytosine coordination chemistry. The assay involved target-induced strand-displacement reaction accompanying dissociation of the chelated Ag(+) in the hairpins and exonuclease III-triggered target recycling. Initially, the added target DNA hybridized with hairpin DNA to disrupt the Ag(I)-coordinated hairpin probe and releases the coordinated Ag(+) ion. Then, the newly formed DNA double-stranded DNA could be cleaved by exonuclease III, and released target HIV DNA, which retriggered the strand-displacement reaction with the hairpin for target recycling, thereby resulting in formation of numerous free Ag(+) ions in the detection cell. The released Ag(+) ions can be readily captured by the negatively charged electrode, and subsequent anodic-stripping voltammetric detection of the captured Ag(+) ions are conducted to form the anodic current for the production of the electronic signal within the applied potential. Under optimal conditions, the exonuclease III-based sensing system exhibited good electrochemical responses for the detection of HIV DNA at a concentration as low as 23 fM.

Changes of lipin1ß expression in gestational diabetes mellitus.

BACKGROUND: Lipin1ß is an adipokine proposed to be associated with insulin resistance (IR). Pregnancy is a physiologic state of progressive IR. The objective of the present study was to investigate the role of lipin1ß in the development of GDM. METHODS: A total of 40 pregnant women (22 normal and 18 with GDM) who delivered healthy infants at full-term (> 37 weeks gestation) were included. The mRNA and protein levels of lipin1ß in adipose tissues were determined by real-time RT-PCR and Western-blot. Plasma glucose, lipids, insulin, and estradiol (E2) levels were measured routinely at fasting state, and HOMA-IR was calculated accordingly. RESULTS: The lipin1ß expression in both mRNA and protein levels in SAT and VAT was lower in GDM patients than controls. Lipin1ß mRNA in VAT was negatively correlated with BMI (r = -0.505, p < 0.05), FINS (r = -0.539, p < 0.05), HOMA-IR (r = -0.574, p < 0.01), TG (r = -0.471, p < 0.05), and E2 (r = -0.564, p < 0.01). Lipin1ß mRNA expression in SAT was similar with VAT. Lipin1ß mRNA was not correlated with body weight gain or blood pressure. These results indicated that the lipin1ß expression in adipose tissues is down-regulated in patients with GDM. CONCLUSIONS: Lipin1ß might play a role in the pathogenesis of insulin resistance in GDM.

Calcium sensing receptor regulating smooth muscle cells proliferation through initiating cystathionine-gamma-lyase/hydrogen sulfide pathway in diabetic rat.

AIMS: Hydrogen sulfide (H2S) inhibits the proliferation of vascular smooth muscle cells (VSMCs). However, how cystathionine-gamma-lyase (CSE), a major enzyme that produces H2S, is regulated remains unknown. Whether calcium-sensing receptor (CaSR) inhibits the proliferation of VSMCs by regulating the endogenous CSE/H2S pathway in diabetic rat has not been previously investigated. METHODS AND RESULTS: The morphological and ultrastructure alterations were tested by transmission electron microscopy, changes in the H2S concentration and the relaxation of the mesenteric secondary artery loop of diabetic rats were determined by Multiskan spectrum microplate spectrophotometer and isometric force transducer. Additionally, the expression levels of CaSR, CSE and Cyclin D1 in the mesenteric arteries of rats were examined by western blotting. The intracellular calcium concentration, the expression of p-CaMK II (phospho-calmodulin kinases II), CSE activity, the concentration of endogenous H2S and the proliferation of cultured VSMCs from rat thoracic aortas were measured by using confocal microscope, western blotting, microplate spectrophotometer, MTT and BrdU, respectively. The VSMC layer thickened, the H2S concentration dropped, the relaxation of the mesenteric secondary artery rings weakened, and the expression of CaSR and CSE decreased whereas the expression of Cyclin D1 increased in diabetic rats compared with the control group. The [Ca(2+)]i of VSMCs increased upon treatment with CaSR agonists (10 µM Calindol and 2.5 mM CaCl2), while it decreased upon administration of calhex231, U73122 and 2-APB. The expression of p-CaMK II and CSE increased upon treatment with CaSR agonists in VSMCs. CSE activity and the endogenous H2S concentration decreased in response to high glucose, while it increased with treatment of CaSR agonists. The proliferation rate increased in response to high glucose, and CaSR agonists or NaHS significantly reversed the proliferation of VSMCs caused by high glucose. CONCLUSIONS: Our results demonstrated that CaSR regulated the endogenous CSE/H2S pathway to inhibit the proliferation of VSMCs in both diabetic and high glucose models.

Hybridization-induced Ag(I) dissociation from an immobilization-free and label-free hairpin DNA: toward a novel electronic monitoring platform.

A novel silver ion (Ag(+))-assisted hairpin DNA through C-Ag(+)-C coordination chemistry was designed for immobilization-free and label-free electrochemical monitoring of human immunodeficiency virus (HIV) DNA on a negatively charged indium-tin oxide electrode, based on hybridization-induced dissociation of silver ions from the hairpin DNA.

Simultaneous analysis of polychlorinated biphenyls and organochlorine pesticides in seawater samples by membrane-assisted solvent extraction combined with gas chromatography-electron capture detector and gas chromatography-tandem mass spectrometry.

A highly efficient and environment-friendly membrane-assisted solvent extraction system combined with gas chromatography-electron capture detector was applied in the simultaneous determination of 17 polychlorinated biphenyls and organochlorine pesticides in seawater samples. Variables affecting extraction efficiency, including extraction solvent used, stirring rate, extraction time, and temperature, were optimized extensively. Under optimal extraction conditions, recoveries between 76.9% and 104.6% in seawater samples were achieved, and relative standard deviation values below 10% were obtained. The limit of detection (signal-to-noise ratio=3) and limit of quantification (signal-to-noise ratio=10) of 17 polychlorinated biphenyls and organochlorine pesticides in seawater ranged from 0.14ngL(-1) to 0.36ngL(-1) and 0.46ngL(-1) to 1.19ngL(-1), respectively. Matrix effects on extraction efficiency were evaluated by comparing with the results obtained using tap water. The extraction effect of developed membrane-assisted solvent extraction method was further demonstrated by gas chromatography-tandem mass spectrometry which can provide structural information of the analytes for more accurate identification, and results identical to those produced by gas chromatography-electron capture detector were obtained. These findings demonstrate the applicability of the developed membrane-assisted solvent extraction determination method for coupling to gas chromatography-electron capture detector or tandem mass spectrometry for determining polychlorinated biphenyls and organochlorine pesticides in seawater samples.

TRB3, up-regulated in kidneys of rats with type1 diabetes, mediates extracellular matrix accumulation in vivo and in vitro.

AIMS: Fibrosis is the final disorder of most chronic kidney disease including diabetic nephropathy (DN), but the mechanisms are not fully understood. The present study aims to determine whether TRB3 participates in fibrogenesis in DN. METHODS: Type1 diabetes was induced in male Wistar rats via intraperitoneal injection of streptozotocin (STZ). The expression of TRB3 and extracellular matrix (ECM) protein collagen I and fibronectin was investigated in kidneys of rats with diabetes and NRK-52E cells (a rat proximal tubular cell line) stimulated with albumin-overload. Rats without diabetes and NRK-52E cells without albumin stimulation served as control. Then gene silencing was used to study whether TRB3 participated in accumulation of collagen I and fibronectin in vivo and in vitro. RESULTS: TRB3 is up-regulated in renal tubules of kidneys of rats with diabetes, especially proximal tubules. Albumin-overload can augments TRB3 expression and increase collagen I and fibronectin secretion in NRK-52E cells. Importantly, silencing of TRB3 alleviates collagen I and fibronectin accumulation in kidneys of rats with diabetes and NRK-52E cells induced by albumin-overload. CONCLUSIONS: TRB3 mediates ECM accumulation in kidneys of rats with STZ-induced type1 diabetes and proximal tubular cells induced by albumin-overload, suggesting a potential target for treatment of DN.

Mesenchymal stem cells transplantation ameliorates glomerular injury in streptozotocin-induced diabetic nephropathy in rats via inhibiting oxidative stress.

AIMS: Mesenchymal stem cells (MSCs) have been demonstrated to be protective in diabetic nephropathy (DN) by reducing albuminuria and attenuating glomerular injury. However, the mechanisms remain unclear. The aim of this study was to explore the effects of MSCs on oxidative stress in DN. MATERIALS/METHODS: Streptozotocin-induced diabetic rats received no treatment or treatment with MSCs (2×10(6), via tail vein) for two continuous weeks. Two other control groups received the antioxidant-probucol or insulin. Eight weeks after treatment, physical, biochemical, renal functional and morphological parameters were measured. Glomerular mesangial cells were cultured for the in vitro experiment. RESULTS: Green fluorescent protein-labeled MSCs were only detected around the glomeruli and near vessels in the kidney. MSCs treatment dramatically reduced blood glucose, urinary albumin excretion, creatinine clearance and renal mass index. The glomerulosclerosis as revealed by periodic acid Schiff staining and expression of collagen I and fibronectin was significantly reduced by MSC treatment. Oxidative stress was also markedly inhibited in the MSCs group. Furthermore, the expression of TGF-ß and membrane localization of GLUT1 were also down-regulated by MSCs. MSCs secreted a significant amount of hepatocyte growth factor (HGF). In vitro, MSC conditioned medium inhibited up-regulation of TGF-ß expression stimulated by high glucose and HGF neutralizing antibody blocked the inhibitory effect of MSC conditioned medium. CONCLUSIONS: MSC treatment reduced urinary albumin excretion and ameliorated glomerulosclerosis. The mechanisms underlying these effects involved reduced blood glucose levels and cellular glucose uptake mediated by GLUT1, thus inhibiting oxidative stress.