Resumo Gardenia jasminoides Ellis é um fitoterápico chinês com valor medicinal e econômico, mas seu mecanismo de resposta ao estresse hídrico permanece obscuro. Neste estudo, o “método de potes duplos” foi usado para simular o estresse hídrico de G. jasminoides Ellis e explorar seu mecanismo de resposta fisiológica e transcriptômica. Não encontramos danos significativos aos lipídios da membrana de G. jasminoides Ellis durante o estresse. POD desempenhou um papel antioxidante vital, o enriquecimento KEGG mostrou que metabólitos secundários, como flavonoides, também podem desempenhar um papel antioxidante e PRO desempenhou um ajuste osmótico significativo. Os hormônios endógenos regulam o crescimento e o desenvolvimento de G. jasminoides Ellis e desempenham um papel na transdução de sinal. Entre eles, o estresse hídrico leve é retardado. Ao mesmo tempo, havia 19631, 23693 e 15045 genes diferencialmente expressos de G. jasminoides Ellis nos dias 5, 10 e 15 sob estresse hídrico. Esses genes foram intimamente associados ao proteassoma, endopeptidase, ribossomo, transdução de sinal MAPK e transdução de sinal de hormônio endógeno, interação planta-patógeno e biossíntese de fenilpropanoides e outras vias fisiológicas e metabólicas, que regulam o turnover e transporte de proteínas, o reforço e adesão das paredes celulares, a indução do fechamento estomático, reações alérgicas, reações de defesa, movimentos foliares, entre outros. Também podem absorver os raios ultravioleta para reduzir a geração de radicais livres de oxigênio, alterar a forma de utilização da energia e ajustar a pressão osmótica das células vegetais.

Abstract Gardenia jasminoides Ellis is a Chinese herbal medicine with medicinal and economic value, but its mechanism of response to waterlogging stress remains unclear. In this study, the “double pots method” was used to simulate the waterlogging stress of Gardenia jasminoides Ellis to explore its physiological and transcriptomic response mechanism. We found no significant damage to Gardenia jasminoides Ellis membrane lipid during stress. POD played a vital antioxidant role, KEGG enrichment showed that secondary metabolites such as flavonoids might also play an antioxidant role, and PRO played a significant osmotic adjustment. Endogenous hormones regulate the Gardenia jasminoides Ellis's growth and development and play a role in signal transduction. Among them, light waterlogging stress is delayed. At the same time, there were 19631, 23693, and 15045 differentially expressed genes on the 5th, 10d, and 15d of Gardenia jasminoides Ellis under waterlogging stress. These genes were closely associated with the proteasome, endopeptidase, ribosome, MAPK signal transduction, and endogenous hormone signal transduction, plant-pathogen interaction and phenylpropanoid biosynthesis and other physiological and metabolic pathways, which regulate the turnover and transportation of protein, the reinforcement and adhesion of cell walls, the induction of stomatal closure, allergic reactions, defense reactions, leaf movements and others. It also can absorb ultraviolet rays to reduce the generation of oxygen free radicals, change the way of energy utilization and adjust the osmotic pressure of plant cells.

Abstract Polygonum capitatum (Polygonaceae) exists in rural areas of Southwest China and some other Asian countries. Its aerial part or whole herbal plant has been used as a Miao ethnomedicine against urinary system diseases, infections, pyelonephritis, diarrhea, and so on. The progress in phytochemical research on P. capitatum over the past decades is summarized. Two hundred and three chemical constituents that have been isolated and identified from this plant are included in this article, the chemical structures of which include aliphatic hydrocarbons, aliphatic alcohols, aliphatic ketones, aromatic aldehydes, aliphatic acids, glycerides, triterpenic acids, sterols, anthraquinones, chromones and their glycosides, flavonoids and their glycosides, phenols, phenolic acids and their esters, lignanoids, flavonolignans, phenolic glycosides, alcoholic glycosides, amino acids, nucleobases, amides, alkaloids, alkyl glycosides, volatile oils, and polysaccharides.

Abstract The structural characterization and bioactivities of a polysaccharide from the stalk residue of Pleurotus eryngii were investigated. Firstly the stalk residue of P. eryngii was collected from the top portion of spent mushroom substrate and processed to yield water-soluble polysaccharide. Subsequently this crude polysaccharide was purified by DEAE Sepharose CL-6B ion exchange chromatography and Sepharose CL-6B size-exclusion chromatography. Then its structural features were investigated by gas chromatography (GC), gel permeation chromatography (GPC), methylation analysis, and Fourier Transform Infrared Spectrum (FT-IR). The results showed that it was heteropolysaccharide and mainly composed of glucose (82.4%). The backbone of P-2a mainly consisted of 1, 3-linked (42.7%) and 1, 6-linked (35.5%) glucose residues. Besides, in vitro antioxidant assay showed that P-2a exerted a high scavenging effects on hydroxyl radical and in vitro antitumor assay showed it had a dose-dependent antiproliferative effect against human gastric MGC-803 cancer cells & human epithelial Hela cancer cells. The findings of this study suggested that the polysaccharide extracted from the stalk residue has the similar structure and bioactivities as that from fruit-body of the mushroom. It could be potentially used as a natural source for the development of health-care food.

ABSTRACT The chemical mechanisms involved in indirect plant-mediated interactions between insects and phytopathogenic fungi on the host plant are poorly understood. Fungus-induced changes in the volatile organic compound (VOC) contents of plants need to be elucidated to address this. Here, changes in VOCs in rose (Rosa chinensis Jacq.) leaves infected with rose powdery mildew (Podosphaera pannosa [Wallr.: Fr.[ de Bary) were studied. VOCs were collected from undamaged live leaves of healthy and infected intact rose plants by dynamic headspace adsorption and identified by GC-MS. VOCs were extracted using n-hexane, and 38 chemicals were found to be produced by P. pannosa. A total of 71 VOCs not produced by P. pannosa were produced to different degrees by infected and healthy plants, and 18 of these were produced only by infected plants. Principal component analysis of chromatographic data gave VOC profiles distinguishing between infected and healthy plants. Hexadecanol, octadecanol, tetradecanol, n-butyl hexadecanoate, and n-butyl stearate dominated the VOCs produced by infected plants. These chemicals can be used as markers for detecting mildew-infected rose plants even 4-7 d after infection before symptoms appear. Clear temporal changes in the concentrations of these five chemicals were found. The results improve our understanding of the chemical mechanisms involved in interactions between insects and phytopathogenic fungi.

Abstract Shallow lowland lakes are critical components of the water cycle, providing an essential service function. However, the impacts of microcystin from phytoplankton communities on benthic macroinvertebrate community diversity and structure have seldom been investigated. During 2008-2012, the impacts of water environmental variables on the diversity of macrobenthic communities, including water temperature, pH, dissolved oxygen, transparency, conductivity, the permanganate index, Chlorophyll a, total phosphorus, total nitrogen, and microcystin-LR (L for leucine and R for arginine), were measured in a typical shallow lowland lake, Ge Lake. The results of the present study demonstrated that there were 31 benthic macroinvertebrate taxa in Ge Lake, including 7 oligochaetes, 7 Mollusca, 14 chironomids, and 3 other taxa. Among the macrobenthic taxa in the benthic community, opportunistic taxa such as Limnodrilus hoffmeisteri are present and can occur at greater densities in disturbed habitats. However, a significant reduction/disappearance of sensitive and clean taxa was observed among the benthic macroinvertebrates. Water temperature, dissolved oxygen, conductivity, ammonia nitrogen, transparency and total phosphorus were the main environmental variables influencing macrobenthic community structure, while water temperature, conductivity, ammonia nitrogen, Chlorophyll a, the permanganate index, total phosphorus and total nitrogen were the main factors that influenced macrobenthic community diversity indices (the numbers of taxa, Shannon’s diversity index, Margalef’s richness index, and Pielou’s evenness index). Our results underscore the severity of the effects of human activity on Ge Lake and strongly suggest that restoring the benthic invertebrate community to previous conditions would require the control and reduction of environmental pollutants and nutrients in Ge Lake.

Due to the special climate conditions in the Qingtongxia region, grapes are high in sugar and low in titratable acidity from the stages of ripening. Therefore, the common methods used for determining the maturity of grapes, which depend on the ratio of sugar and titratable acidity in other regions, are inappropriate in Qingtongxia. This research was done in order to seek for a simple and convenient method of determining the optimal harvest time of grapes, further providing some theoretical basis for improving the quality of wine in Qingtongxia. Phenolic contents and some basic physico-chemical parameters of Merlot and Pinot Noir were evaluated during different ripening stages. The results showed that a different harvest time significantly affects the phenolic contents and physico-chemical parameters of Merlot and Pinot Noir. The total contents of anthocyanins in skins and total contents of phenolic in seeds was screen out as two important indexes to evaluate the maturity of polyphenols, in order to better improve the quality of grape and wine.

Due to the special climate conditions in the Qingtongxia region, grapes are high in sugar and low in titratable acidity from the stages of ripening. Therefore, the common methods used for determining the maturity of grapes, which depend on the ratio of sugar and titratable acidity in other regions, are inappropriate in Qingtongxia. This research was done in order to seek for a simple and convenient method of determining the optimal harvest time of grapes, further providing some theoretical basis for improving the quality of wine in Qingtongxia. Phenolic contents and some basic physico-chemical parameters of Merlot and Pinot Noir were evaluated during different ripening stages. The results showed that a different harvest time significantly affects the phenolic contents and physico-chemical parameters of Merlot and Pinot Noir. The total contents of anthocyanins in skins and total contents of phenolic in seeds was screen out as two important indexes to evaluate the maturity of polyphenols, in order to better improve the quality of grape and wine.

ABSTRACT The topmouth gudgeon, Pseudorasbora parva (Temminck & Schlegel, 1842), is a small cyprinid fish that inhabits the littoral zones of freshwater habitats throughout Asia and Europe. It is regarded as an invasive species in Europe, but in its native range, in China, as food and as an environmental bio-indicator. In this study, the embryonic and larval development of P. parva was investigated for the first time. The eggs of P. parva are transparent, adhesive and elliptical. The mean size of fertilized eggs was (1.63×1.31) ± 0.04 mm. From fertilization to hatching, embryonic development could be divided into eight stages and 34 phases, and the incubation period lasted for 109.25 hours at 24 ± 1 °C. Newly hatched larvae were 4.1 ± 3 mm in length, and the yolk absorption was completed within six days after hatching. The first and second swim bladders formed at the third and ninth day, respectively. The pectoral fin formed before the hatching stage, followed by the caudal, dorsal, anal and ventral fin formation after hatching. About 20 days after hatching, the morphology of the fry was similar to the adult fish. These findings provide a basis for determining the complete ontogeny of P. parva, as well as facilitate the management and utilization of this fish.

This study aimed to investigate the efficacy of minimally invasive tonsil surgery for the treatment of obstructive sleep apnea-hypopnea syndrome (OSAHS) in children. Tonsil ablation or turbinate reduction was performed on 49 pediatric patients with OSAHS by minimally invasive tonsil surgery. In order to evaluate the efficacy of surgery, a comparison was conducted between pre-operation and post-operation data in terms of the symptoms, signs and polysomnography test. Total effectiveness rate of the surgery was 83.7%. Subgroup analysis was also performed based on the severity of their conditions: mild, moderate, and severe groups had an effectiveness rate of 90.0, 88.9, and 66.7%, respectively (Hc=6.665, P<0.05). Postoperatively, the apnea-hypopnea index, the minimum oxygen saturation (SaO2), and corresponding symptoms improved compared to pre-operation conditions (P<0.05). Minimally invasive tonsil surgery was a safe and effective method for treating OSAHS in children.

MicroRNAs (miRNAs) play an important role in drug resistance and modulate the efficiency of chemotherapy. A recent study indicated that miR-340 functions as a tumor suppressor in various types of cancer. However, the role of miR-340 in chemotherapy has not been reported yet. In this study, we found that miR-340 enhanced cisplatin (CDDP)-induced cell death. Induction of miR-340-5p expression decreased the IC50 of CDDP and increased the apoptosis of CDDP-resistant MG-63 and Saos-2 cells. Moreover, miR-340-5p decreased the accumulation of MRP1 and MDR1. We further explored the mechanism underlying the promoting effects of miR-340-5p on CDDP-induced cell death. We identified a potential target of miR-340 in the 3′ untranslated region of lysophosphatidic acid acyltransferase (LPAATβ) using the online program Targetscan (http://www.microrna.org). Luciferase reporter assays showed that miR-340 binds to the 3′UTR of LPAATβ. Enforced expression of miR-340-5p decreased the accumulation of LPAATβ in both MG-63 and Saos-2 cells. Silencing LPAATβ decreased the IC50 of CDDP and increased the apoptosis of CDDP-resistant MG-63 and Saos-2 cells, which is consistent with the effect of miR-340-5p on CDDP-induced cell death. Moreover, induced expression of LPAATβ compromised the effects of miR-340-5p on CDDP-induced cell death and accumulation of MRP1 and MDR1. Taken together, our data indicated that miR-340-5p enhanced the sensitivity to CDDP by targeting LPAATβ.

Background Aspergillus niger has the ability to secrete feruloyl esterase. However, for economically viable industrial applications, it is necessary to increase their catalytic activities and/or protein yields to satisfy the increasing needs for feruloyl esterases. Results The gene AnFaeA that encodes a type A feruloyl esterase was successfully expressed in Pichia pastoris by a two-copy engineered yeast. After a screen in shaker flask, a one-copy strain GSKFA3 having the highest feruloyl esterase activity of 2.4 U/mL was obtained. Then, the pPICZaA-AnFaeA plasmid was transformed into GSKFA3 and the transformants were grown on YPDS plates with antibiotic Zeocin. After cultivation, a two-copy strain GSKZaFA20 with the highest feruloyl esterase activity of 15.49 U/mL was obtained. The expressed protein (recombinant AnFaeA) may be a glycoprotein with an apparent molecular weight of 40 kDa. It displayed the maximum activity at pH 6.0 and 50°C, and was stable at a pH range of 4.0-6.5 and at below 45°C. Its activity was not significantly affected by K+, Ca2 +, Mg2 +, Cu2 +, Zn2 +, Mn2 +, Na+ and EDTA, but activated by Fe2 +. The Km and Vmax toward 4-nitrophenyl ferulate were 5.5 mM and 69.0 U/mg, respectively. Conclusions The two-copy strain GSKZaFA20 showed a 4.4-fold increase in extracellular enzyme activity compared with the one-copy strain GSKFA3. Construction of two-copy strain improved secretion of recombinant AnFaeA in P. pastoris.

There has been concern regarding the use of controversial paradigms for repetitive transcranial magnetic stimulation (rTMS) to manage treatment-resistant depression (TRD). This meta-analysis assessed the efficacy of bilateral rTMS compared with unilateral and sham rTMS in patients with TRD. PubMed, Embase, CENTRAL, PsycINFO, Web of Science, EAGLE and NTIS databases were searched to identify relevant studies, and randomized controlled trials (RCTs) on bilateral rTMS for TRD patients were included. The response was defined as the primary outcome, and remission was the secondary outcome. Ten RCTs that included 634 patients met the eligibility criteria. The risk ratio (RRs) of both the primary and secondary outcomes of bilateral rTMS showed non-significant increases compared to unilateral rTMS (RR=1.01, P=0.93; odds ratio [OR]=0.77, P=0.22). Notably, the RR of the primary bilateral rTMS outcome was significantly increased compared to that for sham rTMS (RR=3.43, P=0.0004). The results of our analysis demonstrated that bilateral rTMS was significantly more effective than sham rTMS but not unilateral rTMS in patients with TRD. Thus, bilateral rTMS may not be a useful paradigm for patients with TRD.

Background Lysozyme plays a crucial role in innate immunity with its well-recognized bacteriolytic activity. In this study, the influence of expression parameters (inoculation volume, culture volume, growth time, induction temperature and time, initial pH and methanol concentration) on human lysozyme (HLZ) production in recombinant P. pastoris SMD1168 was investigated through Plackett-Burman (PB) design and response surface methodology (RSM). Results It was revealed that induction temperature, induction time and culture volume had significant influence (P < 0.01) on HLZ expression level, which were elected for further optimization with three-dimensional response surface designs for enhanced HLZ production. The highest lysozyme activity reached 3301 U/mL under optimized conditions (at 23.5°C for 90 h with culture volume of 48 mL) in shake flask, which increased 2.2 fold compared with that achieved with the standard protocol (Invitrogen). When high-cell-density fermentation of the recombinant Pichia pastoris was performed in a 15 L fermenter under optimized conditions, the extracellular lysozyme activity reached 47,680 U/mL. SDS-PAGE analysis of the product demonstrated that HLZ was produced as a single major protein with a molecular weight of approximately 14.7 kDa, consistent with its expected size. Conclusions The results indicated that the optimized culture conditions using PB design and RSM significantly enhanced the expression level of HLZ, and the Pichia expression system for HLZ production was successful and industrially promising.

FUNDAMENTO: Índices de ondas P são marcadores interessantes para prever recorrências de fibrilação atrial (FA) pós ablação. OBJETIVO: Esse estudo avalia o valor dos índices de onda P para prever recorrências após isolamento da veia pulmonar (IVP) em pacientes com fibrilação atrial paroxística. MÉTODOS: Foram selecionados 198 pacientes (57 ± 8 anos, 150 homens) com FA paroxística sintomática refratária a medicamentos submetidos ao IVP em nosso hospital. Um eletrocardiograma de 12 derivações foi utilizado para medir a duração da onda P na derivação II, a força terminal de P (FTP) na derivação V1, o eixo e a dispersão da onda P. RESULTADOS: No acompanhamento de 9 ± 3 meses, as recorrências ocorreram em 60 (30,3%) pacientes. Os pacientes que apresentaram recorrência de FA tiveram maior duração média de onda P (122,9 ± 10,3 versus 104,3 ± 14,2 ms, p < 0,001), maior dispersão da onda P (40,7 ± 1,7 ms vs 36,6 ± 3,2 ms, p < 0,001). A duração da onda P > 125 ms apresenta 60% de sensibilidade, especificidade de 90%, valor preditivo positivo (VPP) de 72% e valor preditivo negativo (VPN) de 83,7%, enquanto a dispersão da onda P > 40 ms tem 78% de sensibilidade, 67% de especificidade, PPV 51% e VPN de 87,6%. 48/66 (72,7%) dos pacientes com FTP < -0,04 mm/segundo vs 12/132 (9%) com FTP > -0,04 mm/segundo tiveram recorrência de FA (p < 0,001). O eixo da onda P não diferiu entre os dois grupos. Na análise multivariada, os índices da onda P não foram independentes do tamanho do átrio esquerdo e da idade. CONCLUSÕES: A duração da onda P > 125 ms, a dispersão da onda P > 40 ms e FTP em V1 < -0,04 mm/sec são bons preditores clínicos das recorrências de FA pós IVP em pacientes com fibrilação atrial paroxística; contudo, eles não foram independentes do tamanho do átrio esquerdo e da idade.

BACKGROUND: P-wave indices are appealing markers for predicting atrial fibrillation (AF) recurrences post ablation. OBJECTIVE: This study evaluates the value of P wave indices to predict recurrences post pulmonary vein isolation (PVI) in patients with paroxysmal AF. METHODS: We selected 198 patients (57 ± 8 years, 150 males) with symptomatic drug-refractory paroxysmal AF undergoing PVI in our hospital. A 12-lead electrocardiogram was used to measure P wave duration in lead II, P wave terminal force (PWTF) in lead V1, P wave axis and dispersion. RESULTS: During a follow-up of 9 ± 3 months, recurrences occurred in 60 (30.3%) patients. The patients that had AF recurrence had longer mean P wave duration (122.9 ± 10.3 vs 104.3 ± 14.2 ms, p < 0.001), larger P wave dispersion (40.7 ± 1.7 ms vs 36.6 ± 3.2 ms, p < 0.001). P wave duration > 125 ms has 60% sensitivity, 90% specificity, positive predictive value (PPV) of 72% and negative predictive value (NPV) of 83.7%, whereas P wave dispersion > 40 ms has 78% sensitivity, 67% specificity, PPV of 51% and NPV of 87.6% 48/66 (72.7%) patients with PWTF < -0.04 mm/second vs12/132(9%) with PWTF > -0.04 mm/second showed recurrence of AF (p < 0.001). P wave axis was not different between two groups. On multivariate analysis, P wave indices were not independent from left atrial size and age. CONCLUSIONS: P wave duration > 125 ms, P wave dispersion > 40 ms and PWTF in V1 < -0.04 mm/sec are good clinical predictors of AF recurrences post PVI in patients with paroxysmal atrial fibrillation; however they were not independent from left atrial size and age.

English grain aphid (EGA, Sitobion avenae Fabricius) is an important pest in wheat (Triticum aestivum L.). To develop EGA-resistant varieties, introducing the desirable genes from related species is regarded as an efficient avenue. In this study, the F1, F2 and F3 plants derived from the cross of EGA-susceptible wheat-Psathyrostachys huashanica Keng ex Kuo amphiploid (PHW-SA, AABBDDNsNs) and EGA-resistant triticale (Zhongsi 828, AABBRR) were analyzed for EGA resistance. Consequently, PHW-SA was moderately susceptible while Zhongsi 828 and their F1 hybrids were immune, suggesting that the resistance is dominant. All the F2 plants showed high resistance or immunity over two years, indicating that EGA resistance genes are more likely carried by the rye (Secale cereale L.) chromosomes rather than the genomes A or B of Zhongsi 828. In the F3 generation, 25 of 239 lines became susceptible. Giemsa C-banding patterns revealed that these F3 lines had 38-40 chromosomes, including complete rye genome except 5R (and 2R in five lines). Genomic in situ hybridization analysis confirmed this result. During meiosis, all the chromosomes formed bivalents. Six bivalents in 20 lines and five bivalents in five lines were characterized from rye. In contrast, their F2 parental lines had 42 chromosomes (21 bivalents), containing 1R-7R of rye. No P. huashanica chromosomes were detected. Therefore, we propose that the rye chromosome 5R may be related to EGA resistance.