Identification of a novel m6A-related lncRNAs signature and immunotherapeutic drug sensitivity in pancreatic adenocarcinoma.

BACKGROUND: Pancreatic adenocarcinoma (PDAC) ranks as the fourth leading cause for cancer-related deaths worldwide. N6-methyladenosine (m6A) and long non-coding RNAs (lncRNAs) are closely related with poor prognosis and immunotherapeutic effect in PDAC. The aim of this study is to construct and validate a m6A-related lncRNAs signature and assess immunotherapeutic drug sensitivity in PDAC. METHODS: RNA-seq data for 178 cases of PDAC patients and 167 cases of normal pancreatic tissue were obtained from TCGA and GTEx databases, respectively. A set of 21 m6A-related genes were downloaded based on the previous report. Co-expression network was conducted to identify m6A-related lncRNAs in PDAC. Cox analyses and least absolute shrinkage and selection operator (Lasso) regression model were used to construct a risk prognosis model. The relationship between signature genes and immune function was explored by single-sample GSEA (ssGSEA). The tumor immune dysfunction and exclusion (TIDE) score and tumor mutation burden (TMB) were utilized to evaluate the response to immunotherapy. Furthermore, the expression levels of 4 m6A-related lncRNAs on PDAC cell lines were measured by the quantitative real-time PCR (qPCR). The drug sensitivity between the high- and low-risk groups was validated using PDAC cell lines by Cell-Counting Kit 8 (CCK8). RESULTS: The risk prognosis model was successfully constructed based on 4 m6A-related lncRNAs, and PDAC patients were divided into the high- and low-risk groups. The overall survival (OS) of the high-risk groups was more unfavorable compared with the low-risk groups. Receiver operating characteristic (ROC) curves demonstrated that the risk prognosis model reasonably predicted the 2-, 3- and 5-year OS of PDAC patients. qPCR analysis confirmed the decreased expression levels of 4 m6A-related lncRNAs in PDAC cells compared to the normal pancreatic cells. Furthermore, CCK8 assay revealed that Phenformin exhibited higher sensitivity in the high-risk groups, while Pyrimethamine exhibited higher sensitivity in the low-risk groups. CONCLUSION: The prognosis of patients with PDAC were well predicted in the risk prognosis model based on m6A-related lncRNAs, and selected immunotherapy drugs have potential values for the treatment of pancreatic cancer.

Small RNA-big impact: exosomal miRNAs in mitochondrial dysfunction in various diseases.

Mitochondria are multitasking organelles involved in maintaining the cell homoeostasis. Beyond its well-established role in cellular bioenergetics, mitochondria also function as signal organelles to propagate various cellular outcomes. However, mitochondria have a self-destructive arsenal of factors driving the development of diseases caused by mitochondrial dysfunction. Extracellular vesicles (EVs), a heterogeneous group of membranous nano-sized vesicles, are present in a variety of bodily fluids. EVs serve as mediators for intercellular interaction. Exosomes are a class of small EVs (30-100 nm) released by most cells. Exosomes carry various cargo including microRNAs (miRNAs), a class of short noncoding RNAs. Recent studies have closely associated exosomal miRNAs with various human diseases, including diseases caused by mitochondrial dysfunction, which are a group of complex multifactorial diseases and have not been comprehensively described. In this review, we first briefly introduce the characteristics of EVs. Then, we focus on possible mechanisms regarding exosome-mitochondria interaction through integrating signalling networks. Moreover, we summarize recent advances in the knowledge of the role of exosomal miRNAs in various diseases, describing how mitochondria are changed in disease status. Finally, we propose future research directions to provide a novel therapeutic strategy that could slow the disease progress mediated by mitochondrial dysfunction.

The Phase Inversion Mechanism of the pH-Sensitive Reversible Invert Emulsion.

Reversible emulsification drilling fluids can achieve conversion between oil-based drilling fluids and water-based drilling fluids at different stages of drilling and completion, combining the advantages of both to achieve the desired drilling and completion effects. The foundation of reversible emulsion drilling fluids lies in reversible emulsions, and the core of a reversible emulsion is the reversible emulsifier. In this study, we prepared a reversible emulsifier, DMOB(N,N-dimethyl-N'-oleic acid-1,4-butanediamine), and investigated the reversible phase inversion process of reversible emulsions, including the changes in the reversible emulsifier (HLB) and its distribution at the oil-water interface (zeta potential). From the perspective of the acid-alkali response mechanism of reversible emulsifiers, we explored the reversible phase inversion mechanism of reversible emulsions and reversible emulsification drilling fluids. It was revealed that the reversible phase inversion of emulsions could be achieved by adjusting the pH of the emulsion system. Then the proportion of ionic surfactants changed in the oil-water interface and subsequently raised/lowered the HLB value of the composite emulsifier at the oil-water interface, leading to reversible phase inversion of the emulsion. The introduction of organic clays into reversible emulsification drilling fluid can affect the reversible conversion performance of the drilling fluids at the oil-water interface. Thus, we also investigated the influence of organic clays on reversible emulsions. It was demonstrated that a dosage of organic clay of ≤2.50 g/100 mL could maintain the reversible phase inversion performance of reversible emulsions. By analyzing the microstructure of the emulsion and the complex oil-water interface, we revealed the mechanism of the influence of organic clay on the reversible emulsion. Organic clay distributed at the oil-water interface not only formed a complex emulsifier with surfactants, but also affected the microstructure of the emulsion, resulting in a difficult acid-induced phase transition, an easy alkali-induced phase transition, and improved overall stability.

Evaluation of composting parameters, technologies and maturity indexes for aerobic manure composting: A meta-analysis.

Aerobic composting is an efficient method to recover nutrients from animal manure. However, there is considerable variability in the management and maturity criteria used across studies, and a systematic meta-analysis focused on compost maturity is currently lacking. This study investigated the optimal range of startup parameters and practical criteria for manure composting maturity, as well as the effectiveness of in situ technologies in enhancing composting maturity. Most maturity indexes were associated with composting GI, making it an ideal tool for evaluating the maturity of manure composts. GI increased with declined final C/N and (Final C/N)/(Initial C/N) (P < 0.01), and therefore a maturity assessment standard for animal manure composting was proposed: a mature compost has a C/N ratio ≤23 and a GI ≥70, while a highly mature compost has a GI ≥90 and preferably (Final C/N)/(Initial C/N) ≤0.8. Meta-analysis demonstrated that C/N ratio regulation, microbial inoculation and adding biochar and magnesium-phosphate salts are effective strategies for improving compost maturity. Specifically, a greater reduction in the C/N ratio during the composting process is beneficial for improving the maturity of compost product. The optimal startup parameters for composting have been determined, recommending an initial C/N ratio of 20-30 and an initial pH of 6.5-8.5. An initial C/N ratio of 26 was identified as the most suitable for promoting compost degradation and microorganism activity. The present results promoted a composting strategy for producing high-quality compost.

Study on the Properties Changes of Reversible Invert Emulsion during the Process from O/W to W/O with Alkali.

The reversible emulsion drilling fluid system combines the advantages of both oil-based and water-based drilling fluids, which can achieve ideal results in different stages of drilling and completion, and the system can be reused to effectively reduce costs. However, the research on reversible emulsions mainly focuses on the development of new reversible emulsifiers, while the specific phase transformation mechanism of reversible emulsion systems is still unclear. In this paper, a stable reversible emulsion was prepared using the reversible emulsifier DMOB as a raw material, and the reversible emulsion performance of the alkali response from the O/W emulsion phase to the W/O emulsion was studied. The microstructure of reversible emulsions was studied by a microscope, a cryogenic transmission electron microscopy, and a laser particle size analyzer. The changes in macroscopic properties of reversible emulsions in the process of alkali conversion were studied by pH, conductivity, demulsification voltage, static stability, viscosity, rheology, and other indicators, and the conversion mechanism of reversible emulsions from O/W emulsion ⟶ bicontinuous structure ⟶ O/W/O emulsion ⟶ W/O emulsion was clarified. The details are as follows: in the first stage, when the amount of NaOH ≤ 0.43 vol.%, the overall particle size of the emulsion decreases first and then increases with the increase in NaOH dosage. In the second stage, when the amount of NaOH was 0.45 vol.%, a double continuous structure was formed inside the emulsion. In the third stage, when the amount of NaOH is 0.48 vol.%, the O/W/O emulsion is formed, and with the increase in stirring time, the internal oil droplets gradually accumulate and are discharged from the water droplets, and finally, the W/O emulsion is formed. In the fourth stage, when the dosage of 0.50 vol.% ≤ NaOH ≤ 5.00 vol.%, the W/O emulsion was formed, and with the increase of NaOH dosage, the structure and compactness between water droplets increased first and then decreased. In the whole process, with the increase in the amount of NaOH solution, the total particle size of the emulsion first decreased and then increased.

Extracellular vesicles for ischemia/reperfusion injury-induced acute kidney injury: a systematic review and meta-analysis of data from animal models.

BACKGROUND: Acute kidney injury (AKI) induced by ischemia/reperfusion injury significantly contribute to the burden of end-stage renal disease. Extracellular vesicles (EVs), especially for stem/progenitor cell-derived EVs (stem/progenitor cell-EVs), have emerged as a promising therapy for ischemia/reperfusion injury-induced AKI. However, their regulatory effects remain poorly understood, and their therapeutic efficiency in clinical trials is controversial. Here, we performed this systematic review and meta-analysis to assess the stem/progenitor cell-EV efficacy in treating ischemia/reperfusion injury-induced AKI in preclinical rodent models. METHODS: A literature search was performed in PubMed, Embase, Scopus, and Web of Science to identify controlled studies about the therapeutic efficiency of stem/progenitor cell-EVs on ischemia/reperfusion injury-induced AKI rodent models. The level of SCr, an indicator of renal function, was regarded as the primary outcome. Meta-regression analysis was used to reveal the influential factors of EV therapy. Sensitivity analysis, cumulative meta-analysis, and assessment of publication bias were also performed in our systematic review and meta-analysis. A standardized mean difference (SMD) was used as the common effect size between stem/progenitor cell-EV-treated and control groups, with values of 0.2, 0.5, 0.8, and 1.0 defined as small, medium, large, and very large effect sizes, respectively. RESULTS: A total of 30 studies with 985 ischemia/reperfusion injury-induced AKI rodent models were included. The pooled results showed that EV injection could lead to a remarkable sCr reduction compared with the control group (SMD, - 3.47; 95%CI, - 4.15 to - 2.80; P < 0.001). Meanwhile, the EV treatment group had lower levels of BUN (SMD, - 3.60; 95%CI, - 4.25 to - 2.94; P < 0.001), indexes for tubular and endothelial injury, renal fibrosis (fibrosis score and α-SMA), renal inflammation (TNF-α, IL-1ß, iNOS, and CD68 + macrophages), but higher levels of indexes for tubular proliferation, angiogenesis-related VEGF, and reactive oxygen species. However, our meta-regression analysis did not identify significant associations between sCr level and cell origins of EVs, injection doses, delivery routes, and therapy and outcome measurement time (all P values > 0.05). Significant publication bias was observed (Egger's test, P < 0.001). CONCLUSION: Stem/progenitor cell-EVs are effective in improving renal function in rodent ischemia/reperfusion injury-induced AKI model. These vesicles may help (i) reduce cell apoptosis and stimulate cell proliferation, (ii) ameliorate inflammatory injury and renal fibrosis, (iii) promote angiogenesis, and (iv) inhibit oxidative stress. However, the current systematic review and meta-analysis did not identify significant influential factors associated with treatment effects. More preclinical studies and thoughtfully designed animal studies are needed in the future.

miR-6869-5p Transported by Plasma Extracellular Vesicles Mediates Renal Tubule Injury and Renin-Angiotensin System Activation in Obesity.

Obesity increases the risk of other diseases, including kidney disease. Local renal tubular renin-angiotensin system (RAS) activation may play a role in obesity-associated kidney disease. Extracellular vehicles (EVs) transmit necessary information in obesity and cause remote organ damage, but the mechanism is unclear. The aim of the study was to investigate whether the plasma EVs cargo miR-6869-5p causes RAS activation and renal tubular damage. We isolated plasma EVs from obese and lean subjects and analyzed differentially-expressed miRNAs using RNA-seq. Then, EVs were co-cultured with human proximal renal tubular epithelial cells (PTECs) in vitro. Immunohistochemical pathology was used to assess the degree of RAS activation and tubule injury in vivo. The tubule damage-associated protein and RAS activation components were detected by Western blot. Obesity led to renal tubule injury and RAS activation in humans and mice. Obese-EVs induce RAS activation and renal tubular injury in PTECs. Importantly, miR-6869-5p-treated PTECs caused RAS activation and renal tubular injury, similar to Obese-EVs. Inhibiting miR-6869-5p decreased RAS activation and renal tubular damage. Our findings indicate that plasma Obese-EVs induce renal tubule injury and RAS activation via miR-6869-5p transport. Thus, miR-6869-5p in plasma Obese-EVs could be a therapeutic target for local RAS activation in obesity-associated kidney disease.

Critical roles of cytokine storm and secondary bacterial infection in acute kidney injury development in COVID-19: A multi-center retrospective cohort study.

Acute kidney injury (AKI) may develop in patients with coronavirus disease 2019 (COVID-19) and is associated with in-hospital death. We investigated the incidence of AKI in 223 hospitalized COVID-19 patients and analyzed the influence factors of AKI. The incidence of cytokine storm syndrome and its correlation with other clinicopathologic variables were also investigated. We retrospectively enrolled adult patients with virologically confirmed COVID-19 who were hospitalized at three hospitals in Wuhan and Guizhou, China between February 13, 2020, and April 8, 2020. We included 124 patients with moderate COVID-19 and 99 with severe COVID-19. AKI was present in 35 (15.7%) patients. The incidence of AKI was 30.3% for severe COVID-19 and 4.0% for moderate COVID-19 (p < 0.001). Furthermore, cytokine storm was found in 30 (13.5%) patients and only found in the severe group. Kidney injury at admission (odds ratio [OR]: 3.132, 95% confidence interval [CI]: 1.150-8.527; p = 0.025), cytokine storm (OR: 4.234, 95% CI: 1.361-13.171; p = 0.013), and acute respiratory distress syndrome (ARDS) (OR: 7.684, 95% CI: 2.622-22.523; p < 0.001) were influence factors of AKI. Seventeen (48.6%) patients who received invasive mechanical ventilation developed AKI, of whom 64.7% (11/17) died. Up to 86.7% of AKI patients with cytokine storms may develop a secondary bacterial infection. The leukocyte counts were significantly higher in AKI patients with cytokine storm than in those without (13.0 × 109/L, interquartile range [IQR] 11.3 vs. 8.3 × 109/L, IQR 7.5, p = 0.005). Approximately 1/6 patients with COVID-19 eventually develop AKI. Kidney injury at admission, cytokine storm and ARDS are influence factors of AKI. Cytokine storm and secondary bacterial infections may be responsible for AKI development in COVID-19 patients.

ParRouting: An Efficient Area Partition-Based Congestion-Aware Routing Algorithm for NoCs.

Routing algorithms is a key factor that determines the performance of NoC (Networks-on-Chip) systems. Regional congestion awareness routing algorithms have shown great potential in improving the performance of NoC. However, it incurs a significant queuing latency when practitioners use existing regional congestion awareness routing algorithms to make routing decisions, thus degrading the performance of NoC. In this paper, we propose an efficient area partition-based congestion-aware routing algorithm, ParRouting, which aims at increasing the throughput and reducing the latency for NoC systems. First, ParRouting partitions the network into two areas (i.e., edge area and central area.) based on node priorities. Then, for the edge area, ParRouting selects the output node based on different priorities for higher throughput; for the central area, ParRouting selects the node in the low congestion direction as the output node for lower queuing latency. Our experimental results indicate that ParRouting achieves a 53.4% reduction in packet average latency over SPLASH -2 ocean application and improves the saturated throughput by up to 38.81% over a synthetic traffic pattern for an NoC system, compared to existing routing algorithms.

Potential role of extracellular vesicles in the pathophysiology of glomerular diseases.

Extracellular vesicles (EVs) are membrane-bound vesicles released by most cells and are found in diverse biological fluids. The release of EVs provides a new mechanism for intercellular communication, allowing cells to transfer their functional cargoes to target cells. Glomerular diseases account for a large proportion of end-stage renal disease (ESRD) worldwide. In recent years, an increasing number of research groups have focused their effort on identifying the functional role of EVs in renal diseases. However, the involvement of EVs in the pathophysiology of glomerular diseases has not been comprehensively described and discussed. In this review, we first briefly introduce the characteristics of EVs. Then, we describe the involvement of EVs in the mechanisms underlying glomerular diseases, including immunological and fibrotic processes. We also discuss what functions EVs derived from different kidney cells have in glomerular diseases and how EVs exert their effects through different signaling pathways. Furthermore, we summarize recent advances in the knowledge of EV involvement in the pathogenesis of various glomerular diseases. Finally, we propose future research directions for identifying better management strategies for glomerular diseases.

Effect of hybrid teaching incorporating problem-based learning on student performance in pathophysiology.

OBJECTIVE: To compare the effectiveness of traditional and hybrid teaching strategies in pathophysiology and to conduct a survey of students' opinions about the hybrid teaching strategy. METHODS: A hybrid pathophysiology course was developed by combining traditional lectures, case- or problem-based learning, group discussion and several quizzes. A total of 167 students were assigned to the hybrid teaching group and 118 students assigned to the traditional lecture group. RESULTS: Compared with students who received traditional lectures, no students in the hybrid teaching class failed the final examination. The percentage of students with high scores was significantly higher in the hybrid teaching class. In addition, 73.7% of students in the hybrid teaching class expressed substantial interest in pathophysiology during the course, and 83% of these students felt they had received essential training and acquired the ability to solve clinical case problems. CONCLUSION: The hybrid teaching strategy is an advanced approach that encourages students to actively learn teaching materials and solve practical clinical problems, and that promotes student interest in pathophysiology.

Synthesis of a well-dispersed CaFe2O4/g-C3N4/CNT composite towards the degradation of toxic water pollutants under visible light.

Herein, we fabricated a ternary photocatalyst composed of CaFe2O4, multiwalled carbon nanotubes (CNTs) and graphitic carbon nitride (g-C3N4) via a simple hydrothermal route. CaFe2O4 acted as a photosensitizer medium and the CNT acted as a co-catalyst, which remarkably enhanced the photocatalytic performances of g-C3N4 towards the degradation of hexavalent chromium (Cr(vi)) and the antibiotic tetracycline (TC) under visible light irradiation. To investigate the morphological and topological features of the photocatalyst, field-emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM) analyses were performed. The surface properties and oxidation state of the CaFe2O4/g-C3N4/CNT composite were determined by X-ray photoelectron spectroscopy (XPS). The recombination rate of the charge carriers and the band gap values of the as-synthesized catalysts were analyzed by photoluminescence spectroscopy (PL) and diffused reflectance spectroscopy (UV/Vis DRS) studies, respectively. Besides the degradation reactions, the high hydrogen production rate of 1050 µmol h-1 under visible light using the CaFe2O4/g-C3N4/CNT composite loaded with 5 wt% CNT was observed. The superior photocatalytic performances of the CaFe2O4/g-C3N4/CNT composite can be ascribed to the effective heterojunction formed between g-C3N4 and the CaFe2O4 matrix, in which the CNT act as a conducting bridge in the system, promoting the production of photoinduced charge carriers in the semiconductor system. Finally, the plausible photocatalytic mechanism towards the degradation of pollutants and hydrogen production was discussed carefully.

Correction: Synthesis of a well-dispersed CaFe2O4/g-C3N4/CNT composite towards the degradation of toxic water pollutants under visible light.

[This corrects the article DOI: 10.1039/C9RA05005A.].

Inhibition of neurite outgrowth using commercial myelin associated glycoprotein-Fc in neuro-2a cells.

Myelin-associated glycoprotein (MAG) inhibits the growth of neurites from nerve cells. Extraction and purification of MAG require complex operations; therefore, we attempted to determine whether commercially available MAG-Fc can replace endogenous MAG for research purposes. Immunofluorescence using specific antibodies against MAG, Nogo receptor (NgR) and paired immunoglobulin-like receptor B (PirB) was used to determine whether MAG-Fc can be endocytosed by neuro-2a cells. In addition, neurite outgrowth of neuro-2a cells treated with different doses of MAG-Fc was evaluated. Enzyme linked immunosorbent assays were used to measure RhoA activity. Western blot assays were conducted to assess Rho-associated protein kinase (ROCK) phosphorylation. Neuro-2a cells expressed NgR and PirB, and MAG-Fc could be endocytosed by binding to NgR and PirB. This activated intracellular signaling pathways to increase RhoA activity and ROCK phosphorylation, ultimately inhibiting neurite outgrowth. These findings not only verify that MAG-Fc can inhibit the growth of neural neurites by activating RhoA signaling pathways, similarly to endogenous MAG, but also clearly demonstrate that commercial MAG-Fc is suitable for experimental studies of neurite outgrowth.

Attenuation of TRPV1 by AMG-517 after nerve injury promotes peripheral axonal regeneration in rats.

Aims The main objective was to investigate the effects of the transient receptor potential cation channel subfamily V member 1 (TRPV1) on nerve regeneration following sciatic transection injury by functional blockage of TRPV1 using AMG-517, a specific blocker of TRPV1. Methods AMG-517 was injected into the area surrounding ipsilateral lumbar dorsal root ganglia 30 min after unilateral sciatic nerve transection. The number of sciatic axons and the expression of growth-associated protein-43 (GAP-43) and glial fibrillary acidic protein was examined using semithin sections, Western blot, and immunofluorescence analyses. Results Blockage of TRPV1 with AMG-517 markedly promoted axonal regeneration, especially at two weeks after sciatic injury; the number of axons was similar to the uninjured control group. After sciatic nerve transection, expression of glial fibrillary acidic protein was decreased and GAP-43 was increased at the proximal stump. However, the expression of both glial fibrillary acidic protein and GAP-43 increased significantly in AMG-517-treated groups. Conclusions TRPV1 may be an important therapeutic target to promote peripheral nerve regeneration after injury.

The Effect of Botulinum Neurotoxin Serotype a Heavy Chain on the Growth Related Proteins and Neurite Outgrowth after Spinal Cord Injury in Rats.

(1) Background: The botulinum toxin A (BoNT-A) heavy chain (HC) can stimulate the growth of primary motor neurites. (2) Methods: A recombinant BoNT/A HC was injected locally plus interval intrathecal catheter of BoNT/A HC to rats with ipsilateral semi-dissociated lumbar spinal cord injuries (SCIs). First, 2D gel with a silver nitrate stain was applied to detect the general pattern of protein expression. Growth associated protein 43 (GAP-43) and superior cervical ganglion 10 (SCG10) were chosen to represent the altered proteins, based on their molecular weight and pI, and were used to further detect their expression. Meanwhile, the neuronal processes were measured. The measurements of thermal hyperalgesia and grasp power at the ipsilateral hindlimb were used to evaluate spinal sensory and motor function, respectively. (3) Results: The local injection of BoNT/A HC followed by its intrathecal catheter intervally altered the spinal protein expression pattern after an SCI; protein expression was similar to normal levels or displayed a remarkable increase. The changes in the expression and distribution of phosphorylated growth associated protein 43(p-GAP 43) and superior cervical ganglion 10 (SCG 10) indicated that the administration of BoNT/A HC to the SCI significantly amplified the expression of p-GAP43 and SCG10 (p < 0.05). Meanwhile, the positive immunofluorescent staining for both p-GAP43 and SCG10 was mainly present near the rostral aspect of the injury, both in the cytoplasm and the neuronal processes. Moreover, the outgrowth of neurites was stimulated by the BoNT/A HC treatment; this was evident from the increase in neurite length, number of branches and the percentage of cells with neuronal processes. The results from the spinal function tests suggested that the BoNT/A HC did not affect sensation, but had a large role in improving the ipsilateral hindlimb grasp power (p < 0.05). (4) Conclusions: The local injection with the intermittent intrathecal administration of BoNT/A heavy chain to rats with SCI increased the local expression of GAP-43 and SCG 10, which might be affiliated with the regeneration of neuronal processes surrounding the injury, and might also be favorable to the relief of spinal motor dysfunction.

Curcumin alleviates liver oxidative stress in type 1 diabetic rats.

The aim of the present study was to determine the effects of curcumin on antioxidants using a rat model of type 1 diabetes. Seven­week­old male Sprague­Dawley rats were injected with Streptozotocin (STZ) intraperitoneally to induce this model, and then treated with 1.0% curcumin (weight ratio) mixed in their diet for 21 days. The present study included three groups: Control group (NC), diabetic rat model group (DC) and a curcumin treated group (Diab­Cur). The results demonstrated that curcumin treatment markedly decreased the blood glucose levels, plasma malondialdehyde concentration and plasma activity of glutathione peroxidase (GSH­Px) and catalase (CAT); however, it increased the plasma superoxide dismutase (SOD) and insulin levels. Curcumin treatment increased the expression of the CAT, GSH­Px, HO­1 and norvegicus NAD(P)H quinone dehydrogenase 1, and decreased the SOD1 expression, which, led to a diminished oxidative stress status. In addition, curcumin treatment significantly increased the protein expression of Keap1 in the Diab­Cur group when compared with the DC group, decreased cytosolic concentrations of Nrf2 while increasing nuclear accumulation of Nrf2. The results provide evidence that oxidative stress in the STZ­induced diabetic rat model may be attenuated by curcumin via the activation of the Keap1­Nrf2­ARE signaling pathway, as evidenced by a decrease in the blood glucose concentration and an increase in the transcription of several antioxidant genes.

Structural and Functional Interactions between Transient Receptor Potential Vanilloid Subfamily 1 and Botulinum Neurotoxin Serotype A.

BACKGROUND: Botulinum neurotoxins are produced by Clostridium botulinum bacteria. There are eight serologically distinct botulinum neurotoxin isoforms (serotypes A-H). Currently, botulinum neurotoxin serotype A (BoNT/A) is commonly used for the treatment of many disorders, such as hyperactive musculoskeletal disorders, dystonia, and pain. However, the effectiveness of BoNT/A for pain alleviation and the mechanisms that mediate the analgesic effects of BoNT/A remain unclear. To define the antinociceptive mechanisms by which BoNT/A functions, the interactions between BoNT/A and the transient receptor potential vanilloid subfamily 1 (TRPV1) were investigated using immunofluorescence, co-immunoprecipitation, and western blot analysis in primary mouse embryonic dorsal root ganglion neuronal cultures. RESULTS: 1) Three-week-old cultured dorsal root ganglion neurons highly expressed transient TRPV1, synaptic vesicle 2A (SV2A) and synaptosomal-associated protein 25 (SNAP-25). SV2A and SNAP-25 are the binding receptor and target protein, respectively, of BoNT/A. 2) TRPV1 colocalized with both BoNT/A and cleaved SNAP-25 when BoNT/A was added to dorsal root ganglia neuronal cultures. 3) After 24 hours of BoNT/A treatment (1 nmol/l), both TRPV1 and BoNT/A positive bands were detected in western blots of immunoprecipitated pellets. 4) Blocking TRPV1 with a specific antibody decreased the cleavage of SNAP-25 by BoNT/A. CONCLUSION: BoNT/A interacts with TRPV1 both structurally and functionally in cultured mouse embryonic dorsal root ganglion neurons. These results suggest that an alternative mechanism is used by BoNT/A to mediate pain relief.

Blockade of transient receptor potential cation channel subfamily V member 1 promotes regeneration after sciatic nerve injury.

The transient receptor potential cation channel subfamily V member 1 (TRPV1) provides the sensation of pain (nociception). However, it remains unknown whether TRPV1 is activated after peripheral nerve injury, or whether activation of TRPV1 affects neural regeneration. In the present study, we established rat models of unilateral sciatic nerve crush injury, with or without pretreatment with AMG517 (300 mg/kg), a TRPV1 antagonist, injected subcutaneously into the ipsilateral paw 60 minutes before injury. At 1 and 2 weeks after injury, we performed immunofluorescence staining of the sciatic nerve at the center of injury, at 0.3 cm proximal and distal to the injury site, and in the dorsal root ganglia. Our results showed that Wallerian degeneration occurred distal to the injury site, and neurite outgrowth and Schwann cell regeneration occurred proximal to the injury. The number of regenerating myelinated and unmyelinated nerve clusters was greater in the AMG517-pretreated rats than in the vehicle-treated group, most notably 2 weeks after injury. TRPV1 expression in the injured sciatic nerve and ipsilateral dorsal root ganglia was markedly greater than on the contralateral side. Pretreatment with AMG517 blocked this effect. These data indicate that TRPV1 is activated or overexpressed after sciatic nerve crush injury, and that blockade of TRPV1 may accelerate regeneration of the injured sciatic nerve.

Colorimetric detection of kanamycin based on analyte-protected silver nanoparticles and aptamer-selective sensing mechanism.

In this work, a novel colorimetric detection method for kanamycin (Kana), a widely used aminoglycoside antibiotic, has been developed using unmodified silver nanoparticles (AgNPs) as sensing probe. The method is designed based on the finding that the analyte (Kana) can protect AgNPs against salt-induced aggregation, and nucleic acid aptamers can decrease the risk of false positives through an aptamer-selective sensing mechanism. By use of the proposed method, selective quantification of Kana can be achieved over the concentration range from 0.05 to 0.6 µg mL(-1) within 20 min. The detection limit is estimated to be 2.6 ng mL(-1), which is much lower than the allowed maximum residue limit. Further studies also demonstrate the applicability of the proposed method in milk samples, revealing that the method may possess enormous potential for practical detection of Kana in the future.