基于hACE2转导建立SARS-CoV-2 spike假病毒感染病证结合的小鼠病毒肺炎模型

目的建立与评价一种安全、经济、有效的病证结合小鼠新冠病毒感染肺炎模型。方法30只KM种小鼠随机分为空白组、对照组及实验1组、实验2组、实验3组,每组各6只。每天把KM小鼠置于模拟寒湿环境中4 h,将表达hACE2的重组腺相关病毒(pAAV-hACE2)采用改良悬挂法气管内给药转导至肺组织,继以新型冠状病毒(SARS-CoV-2)spike假病毒采用相同的气管内给药方式造模。观察各组小鼠一般情况,检测体重变化、肺指数、血清炎性因子及肺部病理变化。结果免疫组化法显示h ACE2在小鼠肺组织中有效表达;在寒湿环境中给予SARS-CoV-2 spike假病毒后,实验组小鼠表现出类似疫毒犯肺证候的体征;且同样喂养条件和时间下,与空白组和对照组小鼠比较,实验组小鼠出现体重下降;实验1、2、3组小鼠出现肺指数显著增大(P<0.05或P<0.01),血清炎性因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)显著升高(P<0.01),肺组织病理改变明显。结论本研究成功建立了一种安全、经济、有效的病证结合小鼠新冠病毒感染肺炎模型。

The wheat sucrose synthase gene TaSus1 is a determinant of grain number per spike

Some haplotypes of the sucrose synthase gene TaSus1 are associated with thousand-grain weight(TGW)in wheat(Triticum aestivum L.).However,no mutations have been identified within the gene to test this association.The effects of TaSus1 on grain number per spike(GNS)also are largely unknown.Our previous genome-wide association study identified TaSus-A1 as a candidate gene controlling fertile spikelet number per spike(FSN).In the present study,we generated two independent mutants for the three TaSus1 homoeologs by CRISPR/Cas9-mediated genome editing.The triple mutants displayed lower FSN,GNS,grain number per spikelet(GNST),and TGW than wild-type plants.In 306 hexaploid wheat accessions,two single-nucleotide polymorphisms in TaSus-A1 contributed differently to GNS.Introgression of the two alleles into a wheat genetic background confirmed their effects.The alleles differed in geographical distribution among the accessions.

The first factor affecting dryland winter wheat grain yield under various mulching measures: Spike number

Water is the key factor limiting dryland wheat grain yield.Mulching affects crop yield and yield components by affecting soil moisture.Further research is needed to determine the relationships between yield components and soil moisture with yield,and to identify the most important factor affecting grain yield under various mulching measures.A long-term 9-yearifeld experiment in the Loess Plateau of Northwest China was carried out with three treatments:no mulch (CK),plastic mulch (M_(P)) and straw mulch (M_(S)).Yield factors and soil moisture were measured,and the relationships between them were explored by correlation analysis,structural equation modeling and significance analysis.The results showed that compared with CK,the average grain yields of M_(P) and M_(S) increased by 13.0and 10.6%,respectively.The average annual grain yield of the M_(P) treatment was 134 kg ha^(–1) higher than the M_(S) treatment.There were no significant differences in yield components among the three treatments (P<0.05).Soil water storage of the M_(S) treatment was greater than the M_(P) treatment,although the differences were not statistically signifiant.Soil water storage during the summer fallow period (SWSSF) and soil water storage before sowing (SWSS) of M_(S) were significantly higher than in CK,which increased by 38.5 and 13.6%,respectively.The relationship between M_(P) and CK was not statistically significant for SWSSF,but the SWSS in M_(P) was significantly higher than in CK.In terms of soil water storage after harvest (SWSH) and water consumption in the growth period(ET),there were no signi?cant differences among the three treatments.Based on the three analysis methods,we found that spike number and ET were positively correlated with grain yield.However,the relative importance of spike number to yield was the greatest in the M_(P )and M_(S) treatments,while that of ET was the greatest in CK.Suifcient SWSSF could indirectly increase spike number and ET in the three treatments.Based on these results,mulch can improve yield and soil water storage.The most important factor affecting the grain yield of dryland wheat was spike number under mulching,and ET with CK.These findings may help us to understand the main factors influencing dryland wheat grain yield under mulching conditions compared to CK.

Translocation of a fish spike from the pharynx to the thyroid gland:A case report

BACKGROUND A fish spike stuck in the throat is a common ear,nose,and throat(ENT)emergency.However,it is very rare for a fish spike to reach the thyroid tissue through the throat,which is very dangerous and can lead to pharyngeal fistula,cervical abscess,mediastinal abscess,and thyroid abscess.Proper and timely management can help reduce complications,especially in elderly patients.CASE SUMMARY In the case presented here,the causative factor was dentures,but improper management aggravated the condition.In the case presented here,an elderly woman with a history of accidentally swallowing fish bones for 20 d had a sensation of foreign bodies in her throat.Eventually,computed tomography(CT)of the neck showed that the left side of the thyroid gland had a dense shadow in the form of a stripe.CONCLUSION If a fishbone foreign body is not visible during endoscopic examination but the patient has significant symptoms,the surgeon should be aware that the fishbone may be lodged in the thyroid.To avoid a misdiagnosis,ultrasound,CT,and other tests can be used to clarify the diagnosis.T The first step in treating a fish bone in the thyroid gland is to determine the position of the foreign body and the extent of the infection,and to develop a personalized surgical plan for its removal.At the same time,scientific information should be made available to the general public so that people know that if a fish bone is accidentally lodged,they should not force it to be swallowed or be spit out by inducing vomiting,which are incorrect methods and may aggravate the condition or even cause it to migrate outside the cavity,leading to serious complications,as in this reported case.

Spike蛋白介导细胞融合程度定量方法的建立及其在药物筛选中的应用

新型冠状病毒Spike蛋白(S蛋白)与受体ACE2结合介导细胞融合的发生,为进一步探究新冠感染合胞体发生、发展的规律及致病机制,明确关键蛋白靶点,筛选干预合胞体形成的药物,笔者拟构建新冠S蛋白介导的细胞融合程度的定量方法.利用pCDH-CMV-MCS-EF1-Puro、pLV-CMV-MCS-3FLAG-IRES-Bla、S蛋白基因序列、ACE2基因序列、LacZ基因序列、EcoRⅠ限制性内切酶及同源重组酶构建慢病毒穿梭质粒pCDH-S,pCDH-ACE2,pLV-α与pLV-ω,利用该慢病毒质粒分别构建稳定表达S蛋白与ACE2蛋白的293FT和A549细胞系,随后利用pLV-α与pLV-ω慢病毒质粒分别感染上述稳定细胞系,构建出可以定量细胞融合程度的细胞系293FT-S_(1-56),A549-S_(1-56),293FT-ACE2△_(11-41),A549-ACE2△_(11-41).最后采用Western Blot蛋白免疫印迹技术鉴定上述融合定量细胞系中S蛋白与ACE2蛋白的过表达效果,并通过细胞共培养确定融合效果,用β-半乳糖苷酶报告基因检测试剂盒定量细胞融合程度.结果显示,慢病毒载体和稳定细胞系构建成功,时间梯度、转染梯度实验显示该系统具有良好的定量灵敏度,并能用于卡莫司他(Camostat mesylate)和氯硝柳胺(Niclosamide)等药物抗融合效果的定量评价.成功构建的新冠病毒Spike蛋白介导细胞融合程度定量系统,可用于不同S蛋白突变体致融合能力评价和抗融合药物筛选评价,可为进一步的研究奠定技术基础.

Estimating wheat spike-leaf composite indicator(SLI)dynamics by coupling spectral indices and machine learning

The contribution of spike photosynthesis to grain yield(GY)has been overlooked in the accurate spectral prediction of yield.Thus,it’s essential to construct and estimate a yield-related phenotypic trait considering spike photosynthesis.Based on field and spectral reflectance data from 19 wheat cultivars under two nitrogen fertilization conditions in two years,our objectives were to(i)construct a yield-related phenotypic trait(spike–leaf composite indicator,SLI)accounting for the contribution of the spike to photosynthesis,(ii)develop a novel spectral index(enhanced triangle vegetation index,ETVI3)sensitive to SLI,and(iii)establish and evaluate SLI estimation models by integrating spectral indices and machine learning algorithms.The results showed that SLI was sensitive to nitrogen fertilizer and wheat cultivar variation as well as a better predictor of yield than the leaf area index.ETVI3 maintained a strong correlation with SLI throughout the growth stage,whereas the correlations of other spectral indices with SLI were poor after spike emergence.Integrating spectral indices and machine learning algorithms improved the estimation accuracy of SLI,with the most accurate estimates of SLI showing coefficient of determination,root mean square error(RMSE),and relative RMSE values of 0.71,0.047,and 26.93%,respectively.These results provide new insights into the role of fruiting organs for the accurate spectral prediction of GY.This high-throughput SLI estimation approach can be applied for wheat yield prediction at whole growth stages and may be assisted with agronomical practices and variety selection.

A rare natural variation in TaFT-D2 underlies QTss.cas-3D associated with increased total spikelet number per spike in wheat

Total spikelet number per spike(TSS)is a crucial yield component in wheat.Dissecting and characterizing major stable quantitative trait loci(QTL)associated with TSS can significantly enhance the genetic improvement of yield potential.In a previous study,we identified a stable major QTL for TSS,named QTss.cas-3D.In the present study,we conducted fine mapping of QTss.cas-3D,interval to approximately 6.35 Mb,ranging from 105.03 to 111.38 Mb,based on the IWGSC RefSeq v2.1.Through genome resequencing and gene function annotation,we identified TraesCS3D03G0308000(TaFT-D2)as the candidate gene.Phenotypic evaluation with paired near-isogenic lines revealed that this locus predominantly increases kernel number per spike by enhancing TSS and fertile spikelet number per spike,without significantly affecting thousand-kernel weight or tiller number.The presence of the TaFT-D2 allele in the parent P3228,which is rare in nature populations,highlights its potential value.This study provides a valuable gene resource and functional marker for wheat molecular breeding aimed at improving TSS and establishes a foundation for gene functional analysis of TaFT-D2.

Innovative COVID-19 Screening: RT-LAMP Assay for Spike and NSP1 Proteins

Coronavirus disease(COVID-19)is a serious respiratory disease that spreads through the coronavirus globally.It soon became a pandemic after its appearance in 2019 and demanded new techniques for its identification and detection.Owing to this situation,RT-LAMP appears to be a novel method for the identification of COVID-19 because of its vast applications,including cost-effectiveness and time-saving.This research highlights the use of RT-LAMP,a more sensitive test than RT-PCR,for the assessment of SARS-CoV-2,the severe acute respiratory illness.To identify the spike(S)and NSP1 protein using RT-LAMP,170 total samples of coronavirus-suspected patients were served in this research.Health certifications and bioethical considerations were taken into consideration.After the sample was extracted from the patient's swabs,RNA was isolated,extracted,and purified.The response was then run on the RT-LAMP at the ideal temperature,and the outcomes could be observed with the unaided eye as they changed from pink to yellow.It is a simple method of determining if the test is positive or negative.For this purpose,both RT-LAMP and RT-PCR tests are used during these procedures.Genes linked with COVID-19 testing including S,nspl,and ORF are suited to coronavirus testing;they have 100%specificity and low sensitivity,but S has more specificity and sensitivity than nspl and ORF,respectively.Out of the 95 positive samples,89(93.68%)samples yielded favorable outcomes utilizing RT-LAMP,while 55 negative samples yielded 100%positive results.The present research demonstrates that RT-LAMP is less sensitive yet more selective for coronavirus detection.

新型“spike 99”香料的GC/MS检验

目的:对缴获的新型"sp ike 99"香料进行主要有机成分检验。方法:采用GC/MS定性方法对缴获"sp ike 99"香料进行定性分析。结果:"sp ike 99"香料中检出JWH-018,JWH-073等合成大麻素成分。结论:含有这些合成大麻素的"sp ike 99"香料样本,应该被认定为是一种新的"策划药"产品。

皮秒脉冲电场调控大鼠运动皮层神经元spike发放的初步研究

近年来,皮秒脉冲电场诱导的生物电效应逐渐引起了国内外研究学者的关注。现有细胞实验已经初步证实,皮秒脉冲电场对神经元细胞及其动作电位有一定影响。然而,目前国内外尚未见皮秒脉冲对神经元spike发放影响的相关研究报道,皮秒脉冲对动物神经元spike发放的调控效果仍为一个未知数。为此,基于可调式皮秒脉冲发生器（最大峰值电压–1 000 V,半高宽550 ps,上升时间150 ps,重复频率0~10 k Hz可调）,以大鼠运动皮层神经元为实验对象,有效开展了皮秒脉冲对大鼠运动皮层电生理活动影响的研究,初步探讨了皮秒脉冲幅值、重复频率及作用时间等对大鼠运动皮层神经元spike发放的影响。实验发现：大鼠运动皮层神经元spike发放的改变与皮秒脉冲幅值、重复频率及作用时间密切相关。当皮秒脉冲电压幅值绝对值大于1 k V时,神经元spike发放开始出现变化。随着皮秒脉冲重复频率的增大,神经元spike发放有先增强后抑制的趋势。随着皮秒脉冲作用时间的增加,神经元spike发放出现增强或抑制效果的频率响应窗口均提前。

太阳spike辐射中的MHD振荡

我们在1981年5月16日所观测到的典型的微波大爆发的spike辐射中,发现存有～1.4—1.6秒的准周期振荡特征。本文依据MHD波理论,对观测进行了分析讨论,本文认为在日冕圈内外传播着的快磁声波(“腊肠”模)调制了源区的磁场以及电子束的投射角分布,从而影响了ECM不稳定性的增长率,因此而产生了spike辐射中的准周期振荡。另外,本文还对一些有关的物理参数作了定量的估算。

严重急性呼吸综合征冠状病毒Spike蛋白诱发小鼠免疫损伤的实验研究

目的研究严重急性呼吸综合征冠状病毒(SARS-CoV)的Spike蛋白(S蛋白)引起免疫病理损伤的分子机制。方法采用信号转导通路发现者基因芯片筛选相关诱导免疫炎症的信号转导通路基因的表达,通过RT-PCR对基因芯片筛选基因进一步验证并观察信号分子阻断剂干预的影响;将SARS-CoV的S蛋白及信号分子阻断剂分别经气管滴注、尾静脉注射的途径感染BALB/c小鼠,通过免疫组织化学观察肺及免疫器官的病理变化。结果S蛋白诱导了与γ干扰素(IFN-γ)类似的JAK-STAT的信号通路相关基因的表达,RT-PCR证实S蛋白诱导了人支气管上皮细胞的γ干扰素诱导蛋白10(IP-10)基因表达,该作用能够被JAK3抑制剂完全阻断。经气管滴注或尾静脉注射S蛋白均可引起小鼠肺损伤和脾脏损伤。JAK3抑制剂能够抑制S蛋白诱导小鼠肺内IP-10表达,减轻病理损伤。结论JAK-STAT信号转导通路的激活可能在SARS-CoV的S蛋白诱导的以IP-10为代表的炎症级联中起重要作用,该发现可为发展抗SARS病毒诱导的免疫损伤治疗提供新的策略。

猪传染性胃肠炎病毒spike蛋白亲和肽的原核表达

【目的】原核表达猪传染性胃肠炎病毒(transmissible gastroenteritis virus of swine,TGEV)spike蛋白亲和肽(SQHT),检测其病毒亲和性,为建立TGEV诊断方法奠定理论和物质基础。【方法】人工合成TGEV spike蛋白亲和肽基因,亚克隆后将该基因分别插入至原核表达载体pET-32a和pGEX-6p-1中,构建重组质粒pET-32a-SQHT和pGEX-6p-SQHT,对重组质粒进行BamHⅠ单酶切和BamHⅠ/XhoⅠ双酶切及测序鉴定。将重组质粒分别转化至大肠杆菌Rosetta(DE3),用IPTG进行诱导表达,对其表达产物的生物学活性进行检测。【结果】亚克隆获得了150 bp的TGEV spike蛋白亲和肽基因。成功构建了重组质粒pET-32a-SQHT和pGEX-6p-SQHT,并诱导表达出重组蛋白TRX-SQHT和GST-SQHT,其分子质量分别为25和31 ku。Western blot分析表明,2种重组蛋白与TGEV病毒粒子具有良好的亲和性;Dot-ELISA分析表明,2种重组蛋白与TGEV病毒粒子的最低结合滴度TCID50为5×102 mL-1。特异性试验表明,重组蛋白TRX-SQHT和GST-SQHT均可与TGEV产生特异性结合,而不与猪流行性腹泻病毒(PEDV)和猪轮状病毒(PoRV)结合。【结论】使用原核细胞成功表达了TGEV spike蛋白亲和肽,表达的重组蛋白具有很好的TGEV特异亲和性。

基于SPIKES沟通模式的病情告知在安宁疗护中的应用研究

目的探讨基于SPIKES沟通模式的病情告知在安宁疗护工作中对患者正性、负性情绪及死亡态度的影响。方法选取某三级甲等综合医院2019年2月-12月收治住院的肿瘤临终患者49例为对照组,2020年1月-2020年11月收治住院的肿瘤临终患者46例为观察组。对照组患者予以常规的病情告知,观察组采用基于SPIKES沟通模式的病情告知。比较两组患者的正性、负性情绪及死亡态度。结果实行病情告知措施前两组患者的正性、负性情绪及死亡态度评分比较差异无统计学意义(p>0.05);实行病情告知后观察组患者负性情绪评分明显低于对照组(p<0.05),观察组患者在自然接受、趋近接受2个维度评分高于对照组,死亡恐惧及死亡逃避2个维度评分低于对照组,且差异有统计学意义(p<0.05)。结论基于SPIKES沟通模式的病情告知,可以缓解临终患者的负性情绪,降低其对死亡的恐惧、能自然接纳死亡,对安宁疗护工作的顺利开展起到积极作用。

Cytological and Molecular Identification of Alien Chromatin in Giant Spike Wheat Germplasm

Alien chromosomes of twelve giant spike wheat germplasm lines were identified by C-banding, genomic in situ hybridization (GISH), sequence characterized amplified region (SCAR), and random amplified polymorphic DNA (RAPD). All lines showed a chromosome number of 2n = 42, five of them carried both a pair of wheat-rye (Triticum aestivum-Secale cereal) 1BL/1RS translocation chromosomes and a pair of Agropyron intermedium (Ai) chromosomes, three carried a pair of Ai chromosomes only, three others carried a pair of 1BL/1RS chromosomes only, and one carried neither 1BL/1BS nor Ai chromosome. Further identification revealed that the identical Ai chromosome in these germplasm lines substituted the chromosome 2D of common wheat (T aestivum L.), designated as 2Ai. The genetic implication and further utilization of 2Ai in wheat improvement were also discussed.

SARS病毒Spike蛋白具有中和活性的抗原表位预测和初步鉴定

目的:预测并合成SARS病毒Spike蛋白中具有中和活性的抗原表位,鉴定其在免疫检测和疫苗设计中的作用。方法:使用二级结构以及基因组对比预测方法,预测Spike蛋白中具有较高抗原活性的肽段序列并人工合成。用直接ELISA法检测在SARS确诊阳性患者血清和健康人血清之间相应抗体的检出差别,确定预测肽段是否为SARS的抗原肽段。结果:预测并人工合成了具有抗原活性的GEVFNATKFPSVYAW多肽,在ELISA检测中,可检测出SARS阳性血清中的针对性抗体,阳性符合率为71.4%,在健康人血清中不能检测出对应的抗体,阴性符合率为92%。结论:预测并合成了具有SARS病毒Spike蛋白的中和表位抗原,其特异性高于全病毒抗原,为SARS的诊断及疫苗的设计奠定了基础。

基于贝叶斯原理的多维Spike Train分类预测模型

神经元集群编码和spike train分析是神经信息处理的关键问题。该文介绍了一种利用高阶多维泊松模型对spike train进行分类预测的方法,并从spike的强度分布、匹配准确性和集成策略上进行了数学论证。最后利用该方法在大鼠U迷宫实验中选取20组作为训练集进行分类测试,实验结果表明,利用该方法得到的分类准确率在97%左右。

An algorithm for automatic identification of multiple developmental stages of rice spikes based on improved Faster R-CNN

Spike development directly affects the yield and quality of rice. We describe an algorithm for automatically identifying multiple developmental stages of rice spikes(AI-MDSRS) that transforms the automatic identification of multiple developmental stages of rice spikes into the detection of rice spikes of diverse maturity levels. The scales vary greatly in different growth and development stages because rice spikes are dense and small, posing challenges for their effective and accurate detection. We describe a rice spike detection model based on an improved faster regions with convolutional neural network(Faster R-CNN).The model incorporates the following optimization strategies: first, Inception_Res Net-v2 replaces VGG16 as a feature extraction network;second, a feature pyramid network(FPN) replaces single-scale feature maps to fuse with region proposal network(RPN);third, region of interest(Ro I) alignment replaces Ro I pooling, and distance-intersection over union(DIo U) is used as a standard for non-maximum suppression(NMS). The performance of the proposed model was compared with that of the original Faster R-CNN and YOLOv4 models. The mean average precision(m AP) of the rice spike detection model was92.47%, a substantial improvement on the original Faster R-CNN model(with 40.96% m AP) and 3.4%higher than that of the YOLOv4 model, experimentally indicating that the model is more accurate and reliable. The identification results of the model for the heading–flowering, milky maturity, and full maturity stages were within two days of the results of manual observation, fully meeting the needs of agricultural activities.

SPIKES沟通模型在年轻乳腺癌患者病情告知中的应用

近年来,年轻乳腺癌患者疾速递增,约占女性罹患肿瘤的26%,成为年轻女性中死亡率增长最快的癌症之一[1]。癌患者得知病情后容易陷入恐惧和悲观中,影响预后[2],如何将坏消息告诉患者,针对癌症患者制定具有人性化和无害化的告知策略和方式,减少对其不利影响,成为医务人员需要思考的重要问题。

Gene and protein expression profiling analysis of young spike development in large spike wheat germplasms

The wheat grain number per spike （GNPS） is a major yield-limiting factor in wheat-breeding programs. Germplasms with a high GNPS are therefore valuable for increasing wheat yield potential. To investigate the molecular characteristics of young spike development in large-spike wheat germplasms with high GNPS, we performed gene and protein expression profiling analysis with three high-GNPS wheat lines （Pubing 3228, Pubing 3504 and 4844-12） and one Iow-GNPS control variety （Fukuho）. The phenotypic data for the spikes in two growth seasons showed that the GNPS of the three large-spike wheat lines were significantly higher than that of the Fukuho control line. The Affymetrix wheat chip and isobaric tags for relative and absolute quantitation-tandam mass spectrometry （iTRAQ-MS/MS） technology were employed for gene and protein expression profiling analyses of young spike development, respectively, at the floret primordia differentiation stage. A total of 598 differentially expressed transcripts （270 up-regulated and 328 down-regulated） and 280 proteins （122 up- regulated and 158 down-regulated） were identified in the three high-GNPS lines compared with the control line. We found that the expression of some floral development-related genes, including Wknoxlb, the AP2 domain protein kinase and the transcription factor HUA2, were up-regulated in the high-GNPS lines. The expression of the SHEPHERD （SHD） gene was up-regulated at both the transcript and protein levels. Overall, these results suggest that multiple regulatory pathways, including the CLAVATA pathway and the meristem-maintaining KNOX protein pathway, take part in the development of the high-GNPS phenotype in our wheat germplasms.