粉尘螨变应原Der f 36重组蛋白制备及其与Der p 36的交叉反应性研究

目的:制备粉尘螨变应原第36组分(Der f 36)重组蛋白,鉴定其免疫原性并进行生物信息学分析。方法:获得Der f 36核酸序列并进行人工合成,插入pET-28a(+)载体,制备pET-28a(+)-Der f 36质粒,转化至BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 36,SDS-PAGE和Western blot鉴定rDer f 36重组蛋白,IgE-ELISA检测其血清IgE结合率,IgE-ELISA抑制实验检测rDer f 36与rDer p 36的交叉反应性;HNEpC细胞与rDer f 36共孵育24 h后测定细胞因子;生物信息学软件比较Der f 36与Der p 36的理化性质和结构。结果:得到了Der f 36的编码基因,全长690 bp,相对分子质量为25.6 kD;IgE-ELISA鉴定rDer f 36血清IgE结合率为42.1%;IgE-ELISA抑制实验结果显示rDer p 36对rDer f 36的抑制率>50.00%[40.00%(8/20)],平均抑制率为52.98%。HNEpC细胞与rDer f 36共孵育后,IL-6、IL-8、IL-33、IL-25和TSLP表达均高于对照组。生物信息学分析显示Der f 36和Der p 36的序列一致性为77.63%,理化性质较为相似;二级结构分析结果显示二者均含有α螺旋、β转角和无规则卷曲,且无规则卷曲含量最高;C2结构域显示高度重叠(RMSD=0.046)。结论:成功制备了rDer f 36重组蛋白,且具有免疫原性,为粉尘螨变应原单组分诊断及治疗奠定了基础。Der f 36与Der p 36的理化性质、二级结构和三级结构高度相似,决定其具有交叉反应性。

COL1A1突变致Van der Hoeve综合征家系的遗传和表型分析并文献复习

目的对一个常染色体显性遗传的Van der Hoeve综合征家系进行详尽的临床表型分析及基因突变检测,明确该家系的致病基因突变位点及该突变对基因编码的影响。方法对收集到的Van der Hoeve综合征家系进行包括病史、体格检查及辅助检查在内的临床资料的收集及外周血液样本的采集,并对22位家系成员进行外显子组测序以及Sanger测序,利用生物信息学软件分析数据。结果该家系共五代,各代连续发病,且每一代男女均可患病,符合常染色体显性遗传特点。该家系中12例患者均自出生时巩膜即呈蓝色且身材矮小,8例患者有骨折病史,可正常愈合,3例患者考虑有Van der Hoeve综合征所致的听力下降,12例患者的COL1A1基因第17号外显子有一个碱基的缺失(c.1128delT),使第376位后的氨基酸编码改变,在第539位提前结束氨基酸编码,该家系中10例无症状者无此突变。结论该家系患者确定为由COL1A1基因c.1128delT突变导致的Van der Hoeve综合征。

屋尘螨致敏蛋白组分Derp1、Derp2和Derp10检测的临床意义

尘螨是最常见的室内变应原来源之一,诱导人IgE反应最主要的尘螨是屋尘螨和粉尘螨。为了解变应原IgE组分检测的临床诊断意义和作用,该文就Der p1、Der p2和Der p10的研究进展进行综述。

尘螨过敏原Der p 7新的IgE抗原表位鉴定及尘螨交叉反应性的研究

目的:尘螨过敏原Der p 7新的IgE抗原表位鉴定及尘螨交叉反应性的研究。方法:ELISA检测华东地区安徽省合肥市的尘螨过敏患者与尘螨过敏临床相关性较高的重组蛋白Der p 7的IgE结合率;通过ELISA和Dot Blot鉴定屋尘螨过敏原Der p 7的新IgE抗原表位;利用圆二色光谱法检测Der p 7和其突变体的结构及热稳定性;最后用ELISA和ELISA抑制实验鉴定尘螨新的交叉过敏物质。结果:通过ELISA实验检测发现,华东地区的合肥市内尘螨过敏患者体内IgE与Der p 7的结合率为36.2%,与华南地区的广州市(37.4%)和西班牙(34.7%)相近。然而,这一结合率受到地理区域、自然环境和生活方式等多种因素的影响,与华北地区的北京市(19.3%)、乌克兰(22.67%)、意大利(28%)相比明显较低,而与非洲(56%)和日本(超过60%)相比则显著不同。ELISA和Dot Blot鉴定出尘螨重要过敏原Der p 7的两个新IgE抗原结合表位,分别是第36位天冬氨酸和第100位天冬氨酸。用圆二色光谱实验证明了Der p 7的结构稳定性,且突变后结构未发生改变,并检测了其热稳定性特征。最后用ELISA和ELISA抑制实验新发现了两种与尘螨存在交叉过敏的物质(小麦和花生)。结论:基于尘螨过敏原Der p 7与华东地区安徽省合肥市内过敏患者IgE的结合率及新的IgE抗原表位的研究为预防、诊断、治疗过敏性疾病提供理论基础,对开发尘螨过敏原低敏疫苗具有参考价值。发现新的尘螨交叉过敏物质为交叉过敏反应的预防、治疗提供了科学依据。

尘螨过敏原Der p 2线性表位肽及其硝基化产物与IgE结合能力

许多过敏原可以介导Ⅰ型超敏反应,通过与IgE特异性结合,引起过敏症状.过敏原与细胞表面的特异性IgE结合的部分叫做表位,其与IgE的结合能力可以表征过敏原致敏性的强弱.Der p 2是一种重要的屋尘螨过敏原,其线性表位中含有的酪氨酸可被空气中的NO_(2)和O_(3)硝基化,从而影响线性表位与IgE的结合能力.本实验研究了Der p 2的线性表位及其硝基化产物与IgE的结合能力.研究发现,Der p 2的两条表位多肽可以有效地结合IgE,硝基化表位多肽的IgE结合能力显著高于未硝基化的表位多肽,且不同位点的硝基化对于IgE结合能力的增强程度也不同.结果表明,硝基化能够位点特异性地增强Der p 2的致敏性.

ZnSb/Ti_(3)C_(2)T_(x)MXene van der Waals heterojunction for flexible near-infrared photodetector arrays

Two-dimension(2D)van der Waals heterojunction holds essential promise in achieving high-performance flexible near-infrared(NIR)photodetector.Here,we report the successful fabrication of ZnSb/Ti_(3)C_(2)T_(x)MXene based flexible NIR photodetector array via a facile photolithography technology.The single ZnSb/Ti_(3)C_(2)T_(x)photodetector exhibited a high light-to-dark current ratio of 4.98,fast response/recovery time(2.5/1.3 s)and excellent stability due to the tight connection between 2D ZnSb nanoplates and 2D Ti_(3)C_(2)T_(x)MXene nanoflakes,and the formed 2D van der Waals heterojunction.Thin polyethylene terephthalate(PET)substrate enables the ZnSb/Ti_(3)C_(2)T_(x)photodetector withstand bending such that stable photoelectrical properties with non-obvious change were maintained over 5000 bending cycles.Moreover,the ZnSb/Ti_(3)C_(2)T_(x)photodetectors were integrated into a 26×5 device array,realizing a NIR image sensing application.

Optoelectronic Synapses Based on MXene/Violet Phosphorus van der Waals Heterojunctions for Visual‑Olfactory Crossmodal Perception

The crossmodal interaction of different senses,which is an important basis for learning and memory in the human brain,is highly desired to be mimicked at the device level for developing neuromorphic crossmodal perception,but related researches are scarce.Here,we demonstrate an optoelectronic synapse for vision-olfactory crossmodal perception based on MXene/violet phosphorus(VP)van der Waals heterojunctions.Benefiting from the efficient separation and transport of photogenerated carriers facilitated by conductive MXene,the photoelectric responsivity of VP is dramatically enhanced by 7 orders of magnitude,reaching up to 7.7 A W^(−1).Excited by ultraviolet light,multiple synaptic functions,including excitatory postsynaptic currents,pairedpulse facilitation,short/long-term plasticity and“learning-experience”behavior,were demonstrated with a low power consumption.Furthermore,the proposed optoelectronic synapse exhibits distinct synaptic behaviors in different gas environments,enabling it to simulate the interaction of visual and olfactory information for crossmodal perception.This work demonstrates the great potential of VP in optoelectronics and provides a promising platform for applications such as virtual reality and neurorobotics.

High-Quality van der Waals Epitaxial CsPbBr_(3)Film Grown on Monolayer Graphene Covered TiO_(2)for High-Performance Solar Cells

Two-dimensional materials have been widely used to tune the growth and energy-level alignment of perovskites.However,their incomplete passivation and chaotic usage amounts are not conducive to the preparation of highquality perovskite films.Herein,we succeeded in obtaining higher-quality CsPbBr_(3)films by introducing large-area monolayer graphene as a stable physical overlay on top of TiO_(2)substrates.Benefiting from the inert and atomic smooth graphene surface,the CsPbBr_(3)film grown on top by the van der Waal epitaxy has higher crystallinity,improved(100)orientation,and an average domain size of up to 1.22μm.Meanwhile,a strong downward band bending is observed at the graphene/perovskite interface,improving the electron extraction to the electron transport layers(ETL).As a result,perovskite film grown on graphene has lower photoluminescence(PL)intensity,shorter carrier lifetime,and fewer defects.Finally,a photovoltaic device based on epitaxy CsPbBr_(3)film is fabricated,exhibiting power conversion efficiency(PCE)of up to 10.64%and stability over 2000 h in the air.

Mass-spring model for elastic wave propagation in multilayered van der Waals metamaterials

Multilayered van der Waals(vdW)materials have attracted increasing interest because of the manipulability of their superior optical,electrical,thermal,and mechanical properties.A mass-spring model(MSM)for elastic wave propagation in multilayered vdW metamaterials is reported in this paper.Molecular dynamics(MD)simulations are adopted to simulate the propagation of elastic waves in multilayered vdW metamaterials.The results show that the graphene/MoS_(2)metamaterials have an elastic wave bandgap in the terahertz range.The MSM for the multilayered vdW metamaterials is proposed,and the numerical simulation results show that this model can well describe the dispersion and transmission characteristics of the multilayered vdW metamaterials.The MSM can predict elastic wave transmission characteristics in multilayered vdW metamaterials stacked with different two-dimensional(2D)materials.The results presented in this paper offer theoretical help for the vibration reduction of multilayered vdW semiconductors.

重组粉尘螨变应原rDer f 27制备及其活性鉴定

目的:制备粉尘螨变应原第27组分(Der f 27)重组蛋白,鉴定其免疫活性。方法:构建pET-28a(+)-Der f 27质粒,转化至E.coli BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 27。IgE-ELISA和IgE-Western blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率,分别将变应性鼻炎患者PBMC、人支气管上皮细胞BESA-2B与rDer f 27共孵育24 h后测定细胞因子表达;生物信息学软件分析Der f 27的理化性质和结构。结果:成功构建pET-28a(+)-Der f 27质粒并转化至BL21(DE3)细胞,经IPTG诱导表达和纯化后,SDS-PAGE和Western blot在约48 kD处见特异性条带;IgE-ELISA和IgEWestern blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率分别为39.5%和45.5%;与对照组相比,变应性鼻炎患者PBMC与rDer f 27共孵育后,IL-6和IL-8表达升高(P<0.05);BESA-2B细胞与rDer f 27共孵育后,IL-10和TGF-β表达降低,IL-17A和IL-23A表达升高(P<0.05);生物信息学结果显示Der f 27属于丝氨酸蛋白酶抑制物家族,具有此家族通用结构及功能,二级结构主要为α螺旋(42.62%)和无规则卷曲(35.60%)。结论:成功制备了具有较好免疫反应性和免疫原性的重组变应原rDer f 27,并揭示其为变应性鼻炎的重要变应原之一。

Rational design of vitamin C/defective carbon van der Waals heterostructure for enhanced activity,durability and storage stability toward oxygen reduction reaction

Metal-free defective carbon materials with abundant active sites have been widely studied as low-cost and efficient oxygen reduction reaction(ORR)electrocatalysts in metal-air batteries.However,the active sites in defective carbon are easily subjected to serious oxidation or hydroxylation during ORR or storage,leading to rapid degradation of activity.Herein,we design a van der Waals heterostructure comprised of vitamin C(VC)and defective carbon(DC)to not only boost the activity but also enhance the durability and storage stability of the DC-VC electrocatalyst.The formation of VC van der Waals between DC and VC is demonstrated to be an effective strategy to protect the defect active sites from oxidation and hydroxylation degradation,thus significantly enhancing the electrochemical durability and storage anti-aging performance.Moreover,the DC-VC van der Waals can reduce the reaction energy barrier to facilitate the ORR.These findings are also confirmed by operando Fourier transform infrared spectroscopy and density functional theory calculations.It is necessary to mention that the preparation of this DC-VC electrocatalyst can be scaled up,and the ORR performance of the largely produced electrocatalyst is demonstrated to be very consistent.Furthermore,the DC-VC-based aluminum-air batteries display very competitive power density with good performance maintenance.

Phonon resonance modulation in weak van der Waals heterostructures:Controlling thermal transport in graphene-silicon nanoparticle systems

The drive for efficient thermal management has intensified with the miniaturization of electronic devices.This study explores the modulation of phonon transport within graphene by introducing silicon nanoparticles influenced by van der Waals forces.Our approach involves the application of non-equilibrium molecular dynamics to assess thermal conductivity while varying the interaction strength,leading to a noteworthy reduction in thermal conductivity.Furthermore,we observe a distinct attenuation in length-dependent behavior within the graphene-nanoparticles system.Our exploration combines wave packet simulations with phonon transmission calculations,aligning with a comprehensive analysis of the phonon transport regime to unveil the underlying physical mechanisms at play.Lastly,we conduct transient molecular dynamics simulations to investigate interfacial thermal conductance between the nanoparticles and the graphene,revealing an enhanced thermal boundary conductance.This research not only contributes to our understanding of phonon transport but also opens a new degree of freedom for utilizing van der Waals nanoparticle-induced resonance,offering promising avenues for the modulation of thermal properties in advanced materials and enhancing their performance in various technological applications.

Low-Cost and Biodegradable Thermoelectric Devices Based on van der Waals Semiconductors on Paper Substrates

We present a method to fabricate handcrafted thermoelectric devices on standard office paper substrates.The devices are based on thin films of WS_(2),Te,and BP(P-type semiconductors)and TiS_(3)and TiS_(2)(N-type semiconductors),deposited by simply rubbing powder of these materials against paper.The thermoelectric properties of these semiconducting films revealed maximum Seebeck coefficients of(+1.32±0.27)mV K^(-1)and(-0.82±0.15)mV K^(-1)for WS_(2)and TiS_(3),respectively.Additionally,Peltier elements were fabricated by interconnecting the P-and N-type films with graphite electrodes.A thermopower value up to 6.11 mV K^(-1)was obtained when the Peltier element were constructed with three junctions.The findings of this work show proof-of-concept devices to illustrate the potential application of semiconducting van der Waals materials in future thermoelectric power generation as well as temperature sensing for low-cost disposable electronic devices.

空调空气滤网灰尘中尘螨变应原Der f1和Der p1基因的检测

目的应用PCR技术检测空调空气滤网灰尘中尘螨变应原Der f1和Der p1的基因。方法从深圳和北京地区分别收集20台、10台空调过滤网上的灰尘样品,分别提取其生物总基因组DNA作PCR模板,分别设计Der f1和Der p1各两套内外扩增引物分两步进行PCR,通过扩增出目的片段来检测空调空气滤网灰尘中尘螨变应原Der f1和Der p1基因片段,并以深层泥土DNA提取物为阴性对照和以培养的粉尘螨、屋尘螨基因组DNA提取物为阳性对照。PCR产物经电泳后切胶回收目的片段并克隆到T载体上,进行DNA序列分析并与Gen Bank进行同源性比较分析。结果深圳地区20台空调空气滤网上收集的灰尘中均含有粉尘螨和屋尘螨主要变应原Der f1和Der p1基因片段;北京地区10台空调空气滤网上收集的灰尘样品中,只有4份均含有Der f1和Der p1;它们扩增部分的序列与GenBank数据库里相应序列同源性为100%。从阴性对照提取物中未扩增得到目的变应原Der f1和Der p1基因片段,从阳性对照的DNA提取物中分别扩增得到目的变应原Der f1和Der p1基因片段。结论本研究首次应用分子生物学技术证实了空调空气滤网上的灰尘中存在尘螨主要变应原Der f1和Der p1基因,并提示深圳地区空调过滤网灰尘中尘螨可能比北京地区的多,从而为今后防治空调中尘螨引起的变态反应性疾病提供了依据。

粉尘螨过敏患者Der f 1和Der f 2特异性IgE的检测分析

为了解粉尘螨(Dermatophagoides farinae)过敏患者对粉尘螨单组分过敏原Der f 1和Der f 2特异性IgE的水平情况,收集了80例疑似尘螨过敏患者样本,对ImmunoCAP过敏原检测系统和浙江大学迪迅粉尘螨检测试剂盒检测的粉尘螨特异性IgE阳性样本,进一步检测其粉尘螨单组分过敏原Der f 1和Der f 2特异性IgE水平.80例样本中2种方法检测粉尘螨均为阳性的有65例,阳性率为81.3%.65例阳性样本中,Der f 1阳性率为87.7%,Der f 2阳性率为90.8%,二者至少一个为阳性的达96.99%,Der f 2特异性IgE水平显著高于Der f 1.儿童的粉尘螨及其2种单组分过敏原Der f 1和Der f 2特异性IgE水平显著高于成人.综上,Der f 1和Der f 2这2种组分过敏原在我国粉尘螨患者中占据了主导地位,两者特异性IgE水平存在显著差异,后者高于前者,且儿童的敏感性显著高于成人.

DER MORDFALL SKARABUS gyptologie in der Kriminalliteratur des frühen 20. Jahrhunderts-ein auβergewhnliches Beispiel

I. Einleitung 1930 verffentlichte Willard Huntingdon Wright unter dem Pseudonym S.S. van Dine in den USA den Kriminalroman The Scarab Murder Case, der 2001 in DuMont’s Kriminalbibliothek erstmals auch auf deutsch als Der Mordfall Skarabus erschien.1

尘螨1类变应原Derf1和Derp1生物信息学分析

目的:分析尘螨1类变应原的理化特性。方法:Gen Bank检索尘螨1类变应原Der f 1和Der p 1的氨基酸序列,利用生物信息学软件预测分析这两个氨基酸序列的一级结构、疏水性、跨膜区、二级结构及空间结构,并对其可能的功能位点进行预测。结果:尘螨1类变应原Der f 1和Der p 1分别编码321和320个氨基酸,分子量分别为36.4和36.1 ku,理论等电点(p1)分别为5.66和5.65,两蛋白均为亲水性蛋白且可能均无跨膜区,Der f 1的二级结构以不规则卷曲为主(43.93%),而Der p1的二级结构以α螺旋为主(43.75%)。两变应原均由蛋白酶抑制子I 29和木瓜蛋白酶C-端2个结构域组成,且半胱氨酸活性位点、组氨酸活性位点和天冬酰胺活性位点的位置相似。结论:Der f 1和Der p 1的这些特性有助于为螨性过敏性疾病的变应原特异性免疫治疗提供理论支持。

粉尘螨主要变应原基因Der f1和Der f3改组的研究

目的:探索粉尘螨变应原基因Der f1和Der f3改组条件,以期获得粉尘螨变应原融合突变基因。方法:RT-PCR方法扩增粉尘螨变应原Der f 1和Der f3基因,用DnaseⅠ分别酶解Der f 1和Der f3基因10、15、20、25、30 min;酶解相同时间的Der f1和Der f3基因片段两两混合,采用DNAshuffling技术对粉尘螨变应原Der f1和Der f3基因进行重组,琼脂糖凝胶电泳检测重组子。结果:以Der f 2 F和Der f2 R为引物在酶切10、15、20、25、30min均可扩增出清晰条带;以Der f1 F和Der f1 R、Der f1 F和Der f2 R、Der f2 F和Der f1 R为引物在酶切10、15、20、25、30min的模板中均无条带出现;而其他引物组合则在不同酶切时间的模板中均出现条带。结论:通过多种条件的组合,可获得多个粉尘螨变应原基因Der f1和Der f3间的融合基因,为大规模制备高效、低价的哮喘疫苗奠定了基础。

粉尘螨主要变应原基因Der f2和Der f3基因改组的优化

目的优化粉尘螨变应原Derf2和Derf3基因改组条件,以期获得粉尘螨变应原融合突变基因。方法 RT-PCR方法扩增粉尘螨变应原Derf2和Derf3基因,用DnaseⅠ分别酶解Derf2和Derf3基因10、15、20、25、30min;酶解相同时间的Derf2和Derf3基因两两混合,采用DNA shuffling技术对粉尘螨变应原Derf2和Derf3基因进行重组,电泳检测重组子。结果以Derf1F和Derf1 R、Derf2 F和Derf2 R为引物分别在酶切10min、15min、20min、25min均可扩增出清晰条带;以Derf2 F和Der f3 R为引物在酶切10min、15min和30min的模板中亦出现PCR片段。结论通过多种条件的组合,可获得多个粉尘螨变应原Derf2和Derf3基因间的融合基因,为大规模制备高效,低价的哮喘疫苗奠定基础。

屋尘螨Ⅰ类变应原Der p1的体内定位

对过敏性疾病主要过敏原屋尘螨Dermatophagoides pteronyssinusⅠ类变应原(Der p1)进行了抗原定位研究。选用经表达和纯化的Der p1抗原蛋白,按常规方法免疫小鼠,分离血清获抗重组Der p1的抗体(Ⅰ抗),用抗鼠IgG荧光抗体为Ⅱ抗,屋尘螨经石蜡切片,在荧光显微镜下对其特异性变应原的定位进行观察。HE染色显示屋尘螨消化系统占据大部分体腔。组织免疫荧光发现屋尘螨Ⅰ类变应原主要定位于螨的中肠组织及肠内容物,而螨的生殖系统、排泄系统等脏器以及几丁质甲壳均呈阴性反应。据此认为屋尘螨Ⅰ类变应原存在于螨的肠内容物和中肠组织中。