RESUMO Antecedentes: A acupuntura é um tratamento para a dor neuropática, mas seu mecanismo permanece obscuro. Estudos anteriores mostraram que a analgesia foi induzida em ratos com dor neuropática quando o conteúdo de adenosina da medula espinhal aumentou após a eletroacupuntura (EA); no entanto, o mecanismo por trás desse aumento induzido por eletroacupuntura não foi esclarecido. Objetivo: Este estudo teve como objetivo determinar o papel que a ecto-5'-nucleotidase desempenha na analgesia induzida por EA para dor neuropática. Métodos: Realizamos eletroacupuntura no ponto de acupuntura de Zusanli no sétimo dia após estabelecer um modelo de rato de dor neuropática induzida por lesão de constrição crônica. Observamos o limiar de retirada mecânica e o limiar de dor térmica, e detectamos a expressão de ecto-5'-nucleotidase na medula espinhal usando Western blot. Modelos de ratos com lesão de constrição crônica foram injetados intraperitonealmente com α, β-metilenadenosina 5'-difosfato, um inibidor de ecto-5'-nucleotidase, 30 min antes da eletroacupuntura. O conteúdo de adenosina da medula espinhal foi detectado por meio de cromatografia líquida de alta eficiência. Por último, o agonista do receptor de adenosina A1 N6-ciclopentiladenosina foi injetado por via intratecal no edema lombar dos ratos, e a retirada mecânica e os limiares de dor térmica foram reavaliados. Resultados: Analgesia e aumento da expressão de ecto-5'-nucleotidase e conteúdo de adenosina na medula espinhal foram observados 1 h após a eletroacupuntura. α, β-metilenadenosina 5'-difosfato foi capaz de inibir a sobrerregulação do conteúdo de adenosina e analgesia induzida por eletroacupuntura. Após administração de N6-ciclopentiladenosina, a analgesia induzida por eletroacupuntura foi restaurada. Conclusões: Nossos resultados sugerem que a eletroacupuntura em Zusanli pode produzir analgesia em modelos de ratos com lesão de constrição crônica, possivelmente por meio do aumento da expressão de ecto-5'-nucleotidase induzida por eletroacupuntura, levando ao aumento da expressão de adenosina na medula espinhal.

ABSTRACT Background: Acupuncture is a treatment for neuropathic pain, but its mechanism remains unclear. Previous studies showed that analgesia was induced in rats with neuropathic pain when their spinal cord adenosine content increased after electroacupuncture (EA); however, the mechanism behind this electroacupuncture-induced increase has not been clarified. Objective: This study aimed to determine the role that ecto-5’-nucleotidase plays in EA-induced analgesia for neuropathic pain. Methods: We performed electroacupuncture at the Zusanli acupoint on the seventh day after establishing a rat model of neuropathic pain induced through chronic constriction injuries. We observed the mechanical withdrawal threshold and thermal pain threshold and detected the expression of ecto-5’-nucleotidase in the spinal cord using Western blot. Chronic constriction injury rat models were intraperitoneally injected with α,β-methyleneadenosine 5'-diphosphate, an ecto-5’-nucleotidase inhibitor, 30 min before electroacupuncture. The adenosine content of the spinal cord was detected using high-performance liquid chromatography. Lastly, the adenosine A1 receptor agonist N6-cyclopentyladenosine was intrathecally injected into the lumbar swelling of the rats, and the mechanical withdrawal and thermal pain thresholds were reevaluated. Results: Analgesia and increased ecto-5’-nucleotidase expression and adenosine content in the spinal cord were observed 1 h after electroacupuncture. α,β-methyleneadenosine 5'-diphosphate was able to inhibit upregulation of adenosine content and electroacupuncture-induced analgesia. After administration of N6-cyclopentyladenosine, electroacupuncture-induced analgesia was restored. Conclusions: Our results suggest that electroacupuncture at Zusanli can produce analgesia in chronic constriction injury rat models, possibly via the increased ecto-5’-nucleotidase expression induced through electroacupuncture, thus leading to increased adenosine expression in the spinal cord.

RESUMO OBJETIVO O objetivo deste estudo é realizar uma inferência funcional genética baseada na rede de coexpressão (CEN), expandindo o escopo do princípio de “Culpa por Associação” (GBA - Guilt by Association) para prever as funções genéticas do diabetes mellitus tipo 1 (T1DM). MÉTODOS Primeiro, os dados transcritos do T1DM foram recuperados do repositório de dados genômicos para a análise dos genes diferenciais (DEGs), e foi gerada uma CEN diferencial ponderada. A área sob a curva ROC (AUC) foi escolhida para determinar a métrica de desempenho para cada termo de Ontologia Genética (GO). A análise da expressão diferencial identificou 325 DEGs no T1DM, e a análise de coexpressão gerou uma CEN diferencial com aresta de peso >0,8. RESULTADOS Um total de 282 anotações de GO com DEGs >20 foram mantidas para inferência funcional. Ao calcular a pontuação de multifuncionalidade dos genes, a inferência da função genética foi realizada para identificar as funções genéticas ideais para T1DM com base na lista de classificação genética ideal. Considerando um valor de AUC >0,7, foram identificadas seis funções genéticas ideais para a T1DM, tais como a regulação do processo imunológico e da atividade dos receptores. CONCLUSÕES A inferência funcional genética baseada em CEN, ao expandir o princípio de GBA, previu seis funções genéticas ideais para o T1DM. Os resultados podem ser caminhos potenciais para tratamentos terapêuticos ou preventivos do T1DM.

SUMMARY OBJECTIVE This study aimed to propose a co-expression-network (CEN) based gene functional inference by extending the “Guilt by Association” (GBA) principle to predict candidate gene functions for type 1 diabetes mellitus (T1DM). METHODS Firstly, transcriptome data of T1DM were retrieved from the genomics data repository for differentially expressed gene (DEGs) analysis, and a weighted differential CEN was generated. The area under the receiver operating characteristics curve (AUC) was chosen to determine the performance metric for each Gene Ontology (GO) term. Differential expression analysis identified 325 DEGs in T1DM, and co-expression analysis generated a differential CEN of edge weight > 0.8. RESULTS A total of 282 GO annotations with DEGs > 20 remained for functional inference. By calculating the multifunctionality score of genes, gene function inference was performed to identify the optimal gene functions for T1DM based on the optimal ranking gene list. Considering an AUC > 0.7, six optimal gene functions for T1DM were identified, such as regulation of immune system process and receptor activity. CONCLUSIONS CEN-based gene functional inference by extending the GBA principle predicted 6 optimal gene functions for T1DM. The results may be potential paths for therapeutic or preventive treatments of T1DM.

Abstract 4-Coumaric acid: coenzyme A ligase (4CL) is a key enzyme in the phenylpropanoid metabolic pathway that regulates the biosynthesis of lignin and flavonoids. Therefore, the study of 4CL is important to explore the accumulation and regulation of metabolites. This study investigated the role that the 4CL2 gene from Dryopteris fragrans (Df4CL2) plays in the metabolite synthesis. Changes in gene expression, enzyme activity, and the content of lignin and flavonoids were measured in different tissues of tobacco as model plant that was successfully transferred with Df4CL2. Tobacco plants with Df4CL2 (transgenic tobacco, TT) were successfully obtained via the Agrobacterium-transformation method. This TT tended to be thicker and had an earlier flowering period than wild type tobacco (WT). The expression levels of Df4CL2 were higher in the stem, leaf, and root in TT compared to WT. In addition, compared to WT, TT had higher 4CL enzyme activity and higher lignin and flavonoids contents. This suggests that Df4CL2 is involved in the synthesis of lignin and flavonoids in D. fragrans. This research provides important evidence toward understanding the phenylpropanoid metabolic pathway in ferns.

Abstract The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) technology has recently emerged as a powerful genomic editing tool with great potential for crop breeding. However, commonly used protocols for screening of CRISPR/Cas9-induced mutations are laborious, time-consuming, and costly. In the present study we examined the applicability of high resolution melting (HRM) analysis fast screening CRISPR/Cas9-induced mutations in T0 plants and subsequent genotyping of T1 populations. Comparative analysis demonstrated that HRM analysis could identify mutant T0 plants carrying various types of mutation, including single nucleotide substitutions and short insertion/deletions, with false positive/negative rates of 0-2.78%. Furthermore, T1 plants derived from single T0 plants could be correctly genotyped by HRM analysis using WT parents and T0 plants as controls. We hence recommend the adoption of HRM analysis in CRISPR/Cas9 mediated genetic studies and breeding in rice and other crop species.

Non-alcoholic fatty liver disease (NAFLD) is a common disease associated with metabolic syndrome and can lead to life-threatening complications like hepatic carcinoma and cirrhosis. Exenatide, a glucagon-like peptide-1 (GLP-1) receptor agonist antidiabetic drug, has the capacity to overcome insulin resistance and attenuate hepatic steatosis but the specific underlying mechanism is unclear. This study was designed to investigate the underlying molecular mechanisms of exenatide therapy on NAFLD. We used in vivo and in vitro techniques to investigate the protective effects of exenatide on fatty liver via fat mass and obesity associated gene (FTO) in a high-fat (HF) diet-induced NAFLD animal model and related cell culture model. Exenatide significantly decreased body weight, serum glucose, insulin, insulin resistance, serum free fatty acid, triglyceride, total cholesterol, low-density lipoprotein, aspartate aminotransferase, and alanine aminotransferase levels in HF-induced obese rabbits. Histological analysis showed that exenatide significantly reversed HF-induced lipid accumulation and inflammatory changes accompanied by decreased FTO mRNA and protein expression, which were abrogated by PI3K inhibitor LY294002. This study indicated that pharmacological interventions with GLP-1 may represent a promising therapeutic strategy for NAFLD.

Abstract: Background: Tetracentron sinense, distributed mainly in central and southwest China, is a threatened deciduous broad-leaved canopy tree. Presently, little is known about the phenotypic diversity in natural populations of T. sinense, and its fitness and evolutionary potential to the fragmental habitat, which is very important for its conservation. Questions: (1) How is the level of phenotypic diversity in natural populations of T. sinense? (2) Which factors influenced the phenotypic variation? Species study: Tetracentron sinense. Study dates: Between September and October 2014 Methods: 20 phenotypic traits from 11 populations were measured. Nested analysis of variance, multi-comparison, principal components analysis, cluster analysis and correlation analysis were used to study the phenotypic variation and its relation to geo-climate factors. Results: Analysis of nested variance showed significant difference among and within populations, but the phenotypic differentiation coefficient and variance component among populations was higher than that within populations. Variation of phenotypic traits in T. sinense was mainly associated with annual sunshine hours, and showed cline variation along longitude and elevation. 11 provenances of T. sinense were clustered into four groups based on PCA. Conclusions: Phenotypic variation in T. sinense mainly existed among population, and there was moderate level of phenotypic differentiation among populations and low level of phenotypic variation within populations in T. sinense. Autogamy or geitonogamy may be the major factor contributing to the low phenotypic variation and moderate genetic divergence within this species. Suggestions for conservation strategies are provided.

ABSTRACT Gentiana veitchiorum Hemsl., Gentianaceae, a traditional Tibetan medicine, was used for the treatment of liver jaundice with damp-heat pathogen, as well as for headache and chronic pharyngitis. A rapid ultra-performance liquid chromatography, photodiode array detector, quadrupole time-of-flight mass spectrometry method was developed for the fast and accurate identification and quantification of the chemical constituents of G. veitchiorum. In fact, eighteen compounds were detected and identified on the basis of their mass spectra, fragment characteristics and comparison with published data. Especially, the MS fragmentation pathways of iridoid glycosides and flavone C-glycosides were illustrated. Five compounds among them were quantified by UHPLC-PDA, including swertiamarin, gentiopicroside, sweroside, isoorientin, and isovitexin. The proposed method was then validated based on the analyses of linearity, accuracy, precision, and recovery. The overall recoveries for the five analytes ranged from 96.54% to 100.81%, with RSD from 1.05% to 1.82%. In addition, ten batches of G. veitchiorum from different areas were also analyzed. The developed method was rapid and reliable for both identification and quantification of the chemical constituents of G. veitchiorum, especially for simultaneous qualitative and quantitative analysis of iridoid glycosides and flavone C-glycosides.

Sugarcane brown rust disease caused by Puccinia melanocephala is one of the important fungal diseases affecting sugarcane yield around the world. Cultivar resistance is the most appropriate control method for this disease. In this study, 62 introgression lines chosen from the crossing Saccharum officinarum L. cv. Ludashi x Erianthus rockii Yunnan 95-19 were evaluated for brown rust resistance using artificial inoculation. More than 30% of the introgression lines were identified as resistant. Clones Yunye 09-609, Yunye 09-618, Yunye 09-622, Yunye 09-636, Yunye 09-637, Yunye 09-645, Yunye 09-647 and Yunye 09-652 were shown to have high levels of resistance. The resistant lines identified in this study could be used as sources of brown rust resistance in sugarcane breeding programs.

Um eletrodo de carbono vítreo modificado com poli(safranina T) foi utilizado para a determinação simultânea de epinefrina (EP) e ácido úrico (AU) na presença de ácido ascórbico (AA). Correntes eletrocatalíticas aumentadas e potenciais bem separados para EP e AU foram observados. As correntes de pico anódico de EP e AU foram lineares em função das concentrações correspondentes, na faixa de 6,0×10-6-1,0×10-4 mol L-1. Além disso, o eletrodo modificado mostrou boa sensitividade e estabilidade. Resultados satisfatórios foram alcançados para a determinação de EP e AU em soluções de injeção de EP bem como em amostras de urina humana.

A poly(safranine T) modified glassy carbon electrode was used for the simultaneous determination of epinephrine (EP) and uric acid (UA) in the presence of ascorbic acid (AA). Enhanced electrocatalytic currents and well-separated potentials for EP and UA were observed. The anodic peak currents of EP and UA were linear to the corresponding concentrations in the range of 6.0×10-6-1.0×10-4 mol L-1. In addition, the modified electrode showed good sensitivity and stability. Satisfactory results were achieved for the determination of EP and UA in injection solutions of EP and in human urine samples.