基于移动终端的小学英语词汇句型整合课实践——“Do you like pears?”教学实录与评析

本文详细记录＂Do you like pears？＂一课的教学过程,授课教师熟练运用信息技术,通过课前问卷调查提升课堂教学的针对性,创设饱含童趣的情境激发学生学习热情并在情境中有效整合新旧知识,通过有梯度的操练强化学生听、说、认、读能力,结合课文创编新情境实现情感升华,分层拓展练习提升学生的语用能力。

Evaluation of the early defoliation trait and identification of resistance genes through a comprehensive transcriptome analysis in pears

Early defoliation,which usually occurs during summer in pear trees,is gradually becoming a major problem that poses a serious threat to the pear industry in southern China.However,there is no system for evaluating the responses of different cultivars to early defoliation,and our knowledge of the potential molecular regulation of the genes underlying this phenomenon is still limited.In this study,we conducted field investigations of 155 pear accessions to assess their resistance or susceptibility to early defoliation.A total of 126 accessions were found to be susceptible to early defoliation,and only 29 accessions were resistant.Among them,19 resistant accessions belong to the sand pear species(Pyrus pyrifolia).To identify the resistance genes related to early defoliation,the healthy and diseased samples of two sand pear accessions,namely,the resistant early defoliation accession‘Whasan’and the susceptible early defoliation accession‘Cuiguan’,were used to perform RNA sequencing.Compared with‘Cuiguan’,a total of 444 genes were uniquely differentially expressed in‘Whasan’.Combined with GO and KEGG enrichment analyses,we found that early defoliation was closely related to the stress response.Furthermore,a weighted gene co-expression network analysis revealed a high correlation of WRKY and ethylene responsive factor(ERF)transcription factors with early defoliation resistance.This study provides useful resistant germplasm resources and new insights into potentially essential genes that respond to early defoliation in pears,which may facilitate a better understanding of the resistance mechanism and molecular breeding of resistant pear cultivars.

The PcHY5 methylation is associated with anthocyanin biosynthesis and transport in ‘Max Red Bartlett' and ‘Bartlett' pears

The red coloring of pear fruits is mainly caused by anthocyanin accumulation. Red sport, represented by the green pear cultivar ‘Bartlett’(BL) and the red-skinned derivative ‘Max Red Bartlett’(MRB), is an ideal material for studying the molecular mechanism of anthocyanin accumulation in pear. Genetic analysis has previously revealed a quantitative trait locus(QTL) associated with red skin color in MRB. However, the key gene in the QTL and the associated regulatory mechanism remain unknown. In the present study, transcriptomic and methylomic analyses were performed using pear skin for comparisons between BL and MRB. These analyses revealed differential PcHY5 DNA methylation levels between the two cultivars;MRB had lower PcHY5 methylation than BL during fruit development, and PcHY5 was more highly expressed in MRB than in BL. These results indicated that PcHY5 is involved in the variations in skin color between BL and MRB. We further used dual luciferase assays to verify that PcHY5 activates the promoters of the anthocyanin biosynthesis and transport genes PcUFGT, PcGST, PcMYB10 and PcMYB114, confirming that PcHY5 not only regulates anthocyanin biosynthesis but also anthocyanin transport. Furthermore, we analyzed a key differentially methylated site between MRB and BL, and found that it was located in an intronic region of PcHY5. The lower methylation levels in this PcHY5 intron in MRB were associated with red fruit color during development, whereas the higher methylation levels at the same site in BL were associated with green fruit color. Based on the differential expression and methylation patterns in PcHY5 and gene functional verification, we hypothesize that PcHY5, which is regulated by methylation levels, affects anthocyanin biosynthesis and transport to cause the variations in skin color between BL and MRB.

阿司匹林单药抗血小板治疗的缺血性卒中患者复发危险因素分析

目的 探讨使用阿司匹林进行单抗二级预防的缺血性卒中患者复发的危险因素。方法 对200例使用阿司匹林单抗治疗1 a后的轻型和中型缺血性卒中患者进行回顾性的分析,根据患者卒中是否复发分为未复发组和复发组,使用单因素分析和多因素Logistic回归模型分析探讨相关指标是否为复发的高危因素,并利用ROC曲线预测危险因素的临界值。结果 单因素分析显示有统计学意义的危险因素为血小板计数、同型半胱氨酸水平和PEAR1基因rs12041331位点的A等位基因(P <0.05)。多因素分析表明复发的独立危险因素为同型半胱氨酸水平[OR=1.16(1.089~1.240),P <0.001]。采用ROC预测的临界值为8.35μmol/L(灵敏度0.847,特异度0.532)。结论 PEAR1基因rs12041331位点的A等位基因、血小板计数和Hcy水平是导致阿司匹林单抗治疗的缺血性卒中患者复发的危险因素,Hcy水平是导致阿司匹林单抗治疗的缺血性卒中患者复发的独立危险因素,可利用Hcy水平预测阿司匹林单抗治疗的缺血性卒中患者复发的风险。

Genome-wide identification and characterization of the PbrATG family in Pyrus bretschneideri and functional analysis of PbrATG1a in response to Botryosphaeria dothidea

The pear is an economic fruit that is widely planted around the world and is loved by people for its rich nutritional value. Autophagy is a self-protection mechanism in eukaryotes, and its occurrence often accompanied by the degradation of damaged substances in cells and the recycling of nutrients. Autophagy is one of the mechanisms through which plants respond to environmental stress and plays an important role in plant development and stress resistance. Functional studies of autophagy-related genes (ATGs) have been performed on a variety of plant species, but little information is available on the ATG family in pear (Pyrus bretschneideri Rehd). Therefore, we analyzed the evolutionary dynamics and performed a genome-wide characterization of the PbrATG gene family. A total of 28 PbrATG members were identified.Phylogenetic analysis showed that PbrATGs were more closely related to ATGs of European pear and apple. Evolutionary analysis revealed that whole-genome duplication (WGD) and dispersed duplication events were the main driving forces of PbrATG family expansion.Expression analysis of different pear tissues showed that all the genes were expressed in different pear tissues, and different PbrATGs are expressed at different times and in different locations. Moreover, all PbrATGs also responded to different abiotic stresses, especially salt and drought stress, which elicited the highest expression levels. Pear seedlings were subsequently infected with Botryosphaeria dothidea (B.dothidea). The results showed that different PbrATGs had different expression patterns at different infection stages. According to the gene expression data, PbrATG1a was selected as a key autophagy gene for further analysis. Silencing of PbrATG1a reduced the resistance of pear to B. dothidea, which resulted in increased lesions, reactive oxygen species (ROS) contents, antioxidant enzyme activity, and gene expression levels in the silenced pear seedlings after B. dothidea inoculation. In this study, a comprehensive bioinformatic analysis of ATGs was conducted, and the functions of PbrATGs in pear development and in response to stress were elucidated, which laid a foundation for further study of the molecular mechanism of autophagy and a new strategy for pear resistance breeding.

PbrARF4 contributes to calyx shedding of fruitlets in ‘Dangshan Suli’ pear by partly regulating the expression of abscission genes

Fruitlet calyx shedding in pear plants is apparently regulated via numerous pathways that involve both environmental triggers and phytohormones cues such as auxin. In this study, we found at 10 days after full bloom (DAFB) higher levels of indoleacetic acid (IAA) and tryptophan (Trp) in calyx persistence fruitlet (CPF) than calyx shedding fruitlet (CSF) ofDanshan Suli’ pear (Pyrus bretschneideri Rhed.). Consisting with this, the activity of indolealdehyde oxidase (IAAIdO), which promotes IAA synthesis, was remarkably increased, and that of peroxidase(POD), which degrades IAA, dropped markedly in CPF but not in CSF. Further, qRT-PCR results revealed that most of 31 PbrARFs (encoding auxin response factors) in Pyrus bretschneideri were highly expressed in CPF, whereas PbrARF4, PbrARF24 and PbrARF26 were significantly downregulated in CPF vis-a-vis CSF. Phylogenetic analysis revealed that 6 PbrARFs clustered in the group III, where PbrARF4 showed the closest affinity with AtARF1 that promotes organ abscission, indicating a putative role of PbrARF4 in mediating the process of calyx shedding in pear. In fact, the ectopic overexpression of PbrARF4 in Solanum lycopersicum resulted in an earlier-formed and deeper abscission layer (AL) in the transgenic plants, whose calyxes were more prone to wilt at the mature red stage (MR) compared with the control plants (wild-type). More importantly, expression levels of the abscission genes SILS and Sl Cel2 in transgenic plants overexpressing PbrARF4 were significantly upregulated in comparation with the WT, whereas those of Sl BI and Sl TAPG2 were considerably inhibited. Further, PbrJOINTLESS and PbrIDA,the two genes related to calyx shedding in pear, were up-regulated more in CSF than CPF. The findings contribute to a better understanding of PbrARFs involved in fruitlet calyx shedding of pear, which could prove beneficial to improving the quality of pear fruit.

PbGIF1 promoting cell-proliferation in pear fruit is transcriptionally activated by Pb RR1

As a cell proliferation regulator involved in wide biological processes in plants,GRF-INTERACTING FACTOR(GIF)controls different tissues development.However,whether GIF participates in fruit development remains unclear.According to transcriptome data,we identified PbGIF1was highly expressed during fruit development in cytokinins induced parthenocarpy pear.In the present study,the biofunction of PbGIF1 was initially verified.Overexpression of PbGIF1 promoted fruit size of transgenic tomato.The size of flesh fruit was not affected by cell expansion but the cell proliferation was promoted by overexpressing Pb GIF1.The accelerated cell proliferation process was also observed in PbGIF1-overexpressed transgenic pear fruit calli.The transcriptional regulation of cytokinins on PbGIF1 was further confirmed by exogenous CPPU treatments in pear fruitlets.To investigate the underlying mechanism,the cytokinins-responded factor,PbRR1,was further focused on.The results of Yeast-one-hybrid assay suggested that PbRR1 can bind to the promoter sequence of PbGIF1.The transcriptional activation of PbRR1 on PbGIF1 was also confirmed by Dual-Luciferase assays.Taken together,the results showed that cytokinins control pear fruit development via the transcriptional activation of PbGIF1 by PbRR1.

Genomic selection of eight fruit traits in pear

Genomic selection (GS) has the potential to improve selection efficiency and shorten the breeding cycle in fruit tree breeding. In this study,we evaluated the effect of prediction methods, marker density and the training population (TP) size on pear GS for improving its performance and reducing cost. We evaluated GS under two scenarios:(1) five-fold cross-validation in an interspecific pear family;(2) independent validation. Based on the cross-validation scheme, the prediction accuracy (PA) of eight fruit traits varied between 0.33 (fruit core vertical diameter)and 0.65 (stone cell content). Except for single fruit weight, a slightly better prediction accuracy (PA) was observed for the five parametrical methods compared with the two non-parametrical methods. In our TP of 310 individuals, 2 000 single nucleotide polymorphism (SNP) markers were sufficient to make reasonably accurate predictions. PAs for different traits increased by 18.21%-46.98%when the TP size increased from 50to 100, but the increment was smaller (-4.13%-33.91%) when the TP size increased from 200 to 250. For independent validation, the PAs ranged from 0.11 to 0.45 using rrBLUP method. In summary, our results showed that the TP size and SNP numbers had a greater impact on the PA than prediction methods. Furthermore, relatedness among the training and validation sets, and the complexity of traits should be considered when designing a TP to predict the test panel.

E3 ubiquitin ligase PbrATL18 is a positive factor in pear resistance to drought and Colletotrichum fructicola infection

The Arabidopsis Toxicos en Levadura(ATL)protein is a subfamily of the E3 ubiquitin ligases,which exists widely in plants and is extensively involved in plant growth and development.Although the ATL family has been identified in other species,such as Arabidopsis,Oryza sativa,and grapevine,few reports on pear ATL gene families have been reported.In this study,92 PbrATL genes were identified and analyzed from the Pyrus breschneideri genome.Motif analysis and phylogenetic tree generation divided them into nine subgroups,and chromosome localization analysis showed that the 92 PbrATL genes were distributed in 16 of 17 pear chromosomes.Transcriptome data and quantitative real-time polymerase chain reaction(qRT-PCR)experiments demonstrated that PbrATL18,PbrATL41,and PbrATL88 were involved in both pear drought resistance and Colletotrichum fructicola infection.In addition,Arabidopsis thaliana overexpressing PbrATL18 showed greater resistance to drought stress than the wild type(WT),and PbrATL18-silenced pear seedlings showed greater sensitivity to drought and C.fructicola infection than the controls.PbrATL18 regulated plant resistance by regulating chitinase(CHI),phenylalanine ammonia-lyase(PAL),polyphenol oxidase(PPO),catalase(CAT),peroxidase(POD),and superoxide dismutase(SOD)activities.This study provided a reference for further exploring the functions of the PbrATL gene in drought resistance and C.fructicola infection.

A Light and Simplified Branch Bending Method for Young Pear Trees

Aiming at high cost and low efficiency of conventional branch bending method in the modern intensive planting and labor-saving cultivation mode of young pear trees,this paper provides a new branch bending method with wide source of raw materials,cheap price and simple operation,which is also suitable for the management of low-age branches in the process of high grafting and upgrading of traditional big trees.

急性脑梗死患者发生阿司匹林抵抗相关危险因素分析

目的:探讨急性脑梗死患者发生阿司匹林抵抗与相关危险因素的关系。方法:潍坊益都市中心医院2022年10月至2023年5月急性脑梗死患者共358名,患者于入院当日开始口服阿司匹林肠溶片100 mg/d,服用阿司匹林10 d后,根据血小板聚集实验结果将患者分为阿司匹林抵抗(Aspirin Resistance, AR)组和阿司匹林敏感(Aspirin sensitivity, AS)组,分别就AR组和AS组的血小板内皮聚集受体1 (Platelet-endothelial aggregation receptor 1, PEAR1)基因型的分布情况、高血压、2型糖尿病、性别、年龄、吸烟、饮酒等数据进行收集。结果:与AS组患者比较,AR组患者在性别、高血压、2型糖尿病、吸烟史等方面差异有统计学意义(P 0.05);AR组PEAR1野生型GG基因频率低于AS组,差异有统计学意义(P < 0.05);AR组PEAR1纯合突变型AA基因频率高于AS组,差异有统计学意义(P < 0.05)。结论:急性脑梗死患者发生阿司匹林抵抗与患者PEAR1基因的多态性、高血压、2型糖尿病、性别和吸烟等因素有关。

Optimal storage temperature and 1-MCP treatment combinations for different marketing times of Korla Xiang pears

Maintenance of green color is the primary indicator of quality in the market evaluation of Korla Xiang pears at present and can generally be achieved through early harvesting and decreasing the storage temperature, but the fruit quality was reduced by early harvesting, and the decreasing storage temperature increased the risk of chilling injury. The objectives of this study were to determine the optimal storage parameters for different storage times and to find ways to preserve the green skin color of pears. Specifically, we analyzed the effects of the ethylene inhibitor, 1-methylcyclopropene （1-MCP）, combined with low temperature on quality and maintenance of the green color of Korla Xiang pears during storage. We found that 1-MCP and/or low temperature reduced the loss of green color at 20℃ after being removed from cold storage. In addition, 1-MCP significantly inhibited the decline of titratable acid and ascorbic acid but had no significant effect on fruit firmness and total soluble solids. Low temperature with or without 1-MCP inhibited the release of ethylene, inhibited the decline in the stalk preservation index, inhibited the increase in decay rate and weight loss rate during storage, and inhibited the increase in the core browning index after 225 days of storage. Different storage temperatures had different effects on the quality of Korla Xiang pears. Despite inhibiting ethylene release, a storage temperature of-1.5℃ increased the respiration rate. Storage at -1.5℃ caused core browning eady during storage due to chilling injury, whereas at 2℃ core browning occurred late during storage due to senescence. In late storage, 1-MCP had no significant effect on the maintenance of Korla Xiang pear quality at 2℃. Based on these results, we determined the optimal combinations of low temperature and 1-MCP treatment to maintain pear quality while avoiding chilling injury. For different marketing times, the optimal conditions for storage until New Year＇s Day （a storage duration of 90 days） are 2℃ or 1-MCP combined with 2℃. For storage until the Spring Festival （a storage duration of 150 days）, the optimal conditions are 0℃ or 1-MCP combined with 0℃, and for storage until May （a storage duration of 225 days）, the best conditions are 1-MCP combined with -1.5℃.

Risk assessment and ranking of pesticide residues in Chinese pears

The presence of pesticide residues in pears is a serious health concern. This study presents the results from a 2-year investigation （2013-2014） that used gas chromatography, GS/MS and UPLC/MS-MS to measure the levels of 104 pesti- cides in 310 pear samples. In 93.2% of the samples, 43 pesticides were detected, of which the maximum residue levels （MRLs） were exceeded in 2.6% of the samples. Multiple residues （two to eight compounds） were present in 69.7% of the samples; one sample contained nine pesticides and one sample contained 10. Only 6.8% of the samples did not contain residues. To assess the health risks, the pesticide residue data have been combined with daily pear consumption data for children and adult populations. A deterministic model was used to assess the chronic and acute exposures based on the Joint Meeting on Pesticide Residues （JMPR） method. A potential acute risk was demonstrated for children in the case of bifenthrin, which was found to be present at 105.36% of the acute reference dose （ARfD） value. The long- term exposure of the Chinese consumer to pesticide residues through the consumption of raw pears was far below the acceptable daily intake （ADI） criterion. Additionally, the matrix ranking scheme was used to classify risk subgroups of pesticides and pear samples. In general, 95.5% of samples were deemed to be safe and nine pesticides were classified as being of a relatively high risk. The findings indicated that the occurrence of pesticide residues in pears should not be considered a serious public health problem. Nevertheless, a more detailed study is required for vulnerable consumer groups, especially children. Continuous monitoring of pesticides in pears and tighter regulation of pesticide residue standards are recommended.

Effects of 1-methylcyclopropene and modified atmosphere packaging on fruit quality and superficial scald in Yali pears during storage

The Yali pear（Pyrus bretschneideri Rehd.） is susceptible to superficial scald during prolonged cold storage and at shelf life. This study investigated the effects of 1-methylcyclopropene（1-MCP） and modified atmosphere packaging（MAP） on changes of fruit quality and superficial scald during cold storage and at shelf life in Yali pear. Compared with MAP, the combination of MAP and 1-MCP（MAP＋1-MCP） treatment reduced the carbon dioxide and ethylene content inside the packaging bag. The 1-MCP, MAP, and MAP＋1-MCP treatments reduced the superficial scald index, malondialdehyde content, O2^-· production rate and relative conductivity and inhibited the accumulation of α-farnesene and conjugated trienes in the peel. 1-MCP and MAP＋1-MCP treatments maintained a higher phenolic content and enhanced the catalase and superoxide dismutase activities in the fruit, while reduced activities of lipoxygenase and polyphenol oxidase in the peel preceding the onset of superficial scald. Comprehensive analysis indicated that the MAP＋1-MCP treatment is the most effective method tested for improving the quality of Yali pears during cold storage and at shelf life.

Penicillium and patulin distribution in pears contaminated with Penicillium expansum. Determination of patulin in pears by UHPLC-MS/MS

The danger of mycotoxin contamination entering the food supply through post-harvest infection is of perennial concern to food safety experts. To explore the distribution of Penicillium expansum and diffusion of its mycotoxin, patulin, in blue mold-damaged pears, Pyrus bretschneideri Rehd. cv. Yali obtained from markets and orchards in China were artificially inoculated with P. expansum and assayed for patulin accumulation and degree of fungal colonization. The inoculated pears were incubated until the lesions were 5, 10, 20, or 30 mm in diameter. We sampled tissue at a range of distances from the lesion, measured the spread of Penicillium by plate colony-counting methods, and used UHPLC-MS/MS to detect and quantify the patulin concentration. More P. expansum colony-forming units were isolated from pears with a higher degree of decay. Farther from the lesion, the fewer P. expansum colonies were observed, and the lower the patulin content detected. We found a significant difference in the patulin content between samples due to lesion size, and also in tissue sampled 10 mm away from the lesion. In consideration of this finding, to ensure food safety, we recommend that when a blue mold rot lesion on pear is 5, 10, or 20 mm in diameter, 20, 30, and 40 mm beyond the lesion should be removed, respectively. If a lesion surpasses 30 mm in diameter, the whole pear should be thrown away.

Effects of Nitric Oxide on the Quality and Pectin Metabolism of Yali Pears During Cold Storage

The effect of fumigation with 10, 20, and 30 μL L-1 nitric oxide （NO） was investigated to study the effects of NO on the quality of Yali pears during cold storage. The ethylene production, composition of cell walls, and cell-wall-modifying enzyme activities were measured on fruits which were fumigated with NO （20 μL L-1）. The results showed that NO not only reduced the peak value of ethylene production rate, the soluble sugar, soluble solid content, maintained higher firmness, starch, and NO content, but also retarded the degradation of covalent soluble pectin, accumulation of ionic soluble pectin and water soluble pectin. Moreover, NO fumigation decreased the activities of polygalacturonase （PG） and b-galactosidase（b-Gal） and delayed the peak of PG activity of fruits. Therefore, it indicated that NO fumigation could delay the softening and ripening of Yali pears.

Bleeding canker of pears caused by Dickeya fangzhongdai:Symptoms,etiology and biology

Bleeding canker,a devastating disease of pear trees(Pyrus pyrifolia L.),was first reported in the 1970 s in Jiangsu,China and more recently in other provinces in China.Trees infected with bleeding canker pathogen,Dickeya fangzhongdai,develop cankers on the trunks and branches,and a rust-colored mixture of bacterial ooze and tree sap could be seen all over the trunks and branches.In this study,we provided detail descriptions of the symptoms and epidemiology of bleeding canker disease.Based on pathogenic and phenotypic characterizations,we identified the causal agent of bleeding canker of pear as D.fangzhongdai.Dickeya fangzhongdai strains isolated from pear were also pathogenic on Solanum tuberosum,Brassica pekinensis,Lycopersicon esculentum,and Phalaenopsis aphrodite based on artificial inoculation,and the pathogen were more virulent on potato than that of D.solani strain.This study provides new information about this disease and bleeding canker disease of pear.

Effects of 1-Methylcyclopropene on NO Content,NOS Activity,and H_2O_2 Content in Postharvest Suli Pears

Studies were conducted over the effects of 1-methylcycolpropene （1-MCP） treatment on postharvest life of Suli pears （Pyrus bretschneideri Rehd.） stored at room and cold temperatures on nitric oxide （NO）,nitric oxide synthase （NOS） activity,and hydrogen dioxide （H2O2）.Results showed that the 1-MCP treatment had little effect on total soluble solids （TSS） at both room and cold temperatures.1-MCP delayed softening of Suli pear fruits,decreased respiratory rate and H2O2 accumulation,and increased NO and NOS activity at room temperature storage,while the effect of cold temperature storage was relatively inferior.There was a significant positive correlation between NOS activity and NO content.It is concluded that 1-MCP had effects on endogenous NO content and accumulation of H2O2.Similarly,H2O2 acting as a signaling molecule via regulating NO level affects the ripening and senescence of Suli pears.

PEARS课程在儿科急诊护士急救能力培训中的应用

[目的]探讨儿童急症的评估、识别和干预(PEARS)课程对提高儿科急诊护士急救能力的作用。[方法]采用美国心脏协会的PEARS课程对急诊护士进行培训,在培训前对护士进行理论摸底考试,培训后进行理论、技能和仿真案例考核,考核结束后进行课程评价的问卷调查。[结果]培训后护士的理论成绩(88.83分±7.86分)较培训前(76.75分±6.21分)提高(P<0.01);护师组的技能考核和仿真案例考核优秀率高于护士组(P<0.01)。[结论]儿科急诊护士经PEARS课程培训后对急诊患儿的评估、识别和干预能力提高。