Reversible cytoplasmic rearrangements precede wall apposition,hypersensitive cell death and defense-related gene activation in potato/Phytophthora infestans interactions

We used video microscopy techniques as a tool for live examination of the dynamic aspects of plant/fungus interactions. Early, dynamic responses of epidermal midrib cells of leaves from a potato cultivar (Solanum tuberosum L. cv. Datura) carrying resistance gene R1 to Phytophthora infestans (race 1: compatible interaction, race 4: incompatible interaction) were monitored. Similar responses were observed in both types of interaction, ranging from no visible reaction of invaded plant cells to hypersensitive cell death. The overall defense response of each individual cell exhibited a highly dynamic behavior that appeared to be tightly coordinated with the growth of the fungus. Initial localized reactions, including major rearrangements within the cytoplasm, occurred directly at the fungal penetration site, where rapid apposition of autofluorescent material and callose took place. If fungal invasion stopped at this stage, the host cell restored its normal cytoplasmic activity and survived. Hypersensitive cell death occurred only when fungal growth had proceeded to the formation of a clearly identifiable haustorium. In such cases, cytoplasm and nucleus conglomerated around the intracellular fungal structure, followed by a sudden collapse of the whole conglomerate and an instantaneous collapse of the fungal haustorium. Only small quantitative differences between the compatible and incompatible interactions of the two fungal races were observed for these early responses of epidermal cells. In the incompatible interaction, a slightly larger number of epidermal cells responded to fungal attack. More pronounced quantitative differences between compatible and incompatible interactions occurred upon fungal invasion of the mesophyll. These differences in the number of responding cells were not reflected at the level of gene expression: the spatial and temporal activation patterns of two defense-related genes, encoding phenylalanine ammonia-lyase and pathogenesis-related protein 1, were similar in both types of interaction.Dedicated to Professor Peter Sitte, Freiburg, Germany, on the occasion of his 65th birthday     

Ultrastructural comparison of incompatible and compatible interactions in the barley powdery mildew disease

Summary In the powdery mildew disease of barley,Erysiphe graminis f. sp.hordei forms an intimate relationship with compatible hosts, in which haustoria form in epidermal cells with no obvious detrimental effects on the host until late in the infection sequence. In incompatible interactions, by contrast, the deposition of papillae and localized host cell death have been correlated with the cessation of growth byE. g. hordei. With the advent of improved, low temperature methods of sample preparation, we felt that it was useful to reevaluate the structural details of interactions between barley andE. g. hordei by transmission electron microscopy. The haustoria that develop in susceptible barley lines appear highly metabolically active based on the occurrrence of abundant endoplasmic reticulum, Golgi-like cisternae, and vesicles. In comparison, haustoria found in the resistant barley line exhibited varying signs of degradation. A striking clearing of the matrix and loss of cristae were typical early changes in the haustorial mitochondria in incompatible interactions. The absence of distinct endoplasmic reticulum and Golgi-like cisternae, the formation of vacuoles, and the occurrence of a distended sheath were characteristic of intermediate stages of haustorial degeneration. At more advanced stages of degeneration, haustoria were dominated by large vacuoles containing membrane fragments. This process of degeneration was not observed in haustoria ofE. g. hordei developing in the susceptible barley line.Abbreviations b endoplasmic reticulum extension, blebbing - er endoplasmic reticulum - f fibrillar material - g Golgi-like structure - h haustorium - hb haustorial body - hcw haustorial cell wall - hcy haustorial cytoplasm - hf haustorial finger - hocw host cell wall - hocy host cytoplasm - 1 lipid-like droplet - m mitochondrion - mt microtubule - mve multivesicular body - n nucleus - p papilla - ph penetration site of an infection peg - pl plasma membrane - s sheath - sm extrahaustorial membrane - v vacuole - ve vesicle     

幽门螺杆菌根除治疗对慢性胃炎患者肠道菌群的影响及其与血清hsCRP水平的相关性

目的探究幽门螺杆菌(H.pylori)根除治疗对慢性胃炎患者肠道菌群的影响及其与血清超敏C反应蛋白(hsCRP)水平的相关性。方法选择2018年5月至2019年5月我院收治的87例慢性胃炎患者为研究对象,所有患者均采用枸橼酸铋钾胶囊+雷贝拉唑钠肠溶胶囊+克拉霉素片+阿莫西林克拉维酸钾片进行幽门螺杆菌根除治疗,评价所有患者治疗后临床疗效、H.pylori根除率,比较所有患者治疗前后临床症状积分,肠道肠球菌、葡萄球菌、肠杆菌、乳杆菌、双歧杆菌数量及血清hsCRP水平,采用Pearson相关分析所有患者治疗后肠道菌群数量与血清hsCRP水平的相关性。结果入选患者临床治疗有效率为95.40%,H.pylori根除率为90.80%。患者治疗后腹胀、嗳气、腹痛、纳差评分显著低于治疗前(均P<0.05)。治疗后患者肠道肠球菌、葡萄球菌数量显著高于治疗前,而肠杆菌、乳杆菌、双歧杆菌数量显著低于治疗前(均P<0.05)。患者治疗后血清hsCRP水平显著低于治疗前(P<0.05)。患者治疗后肠道肠球菌、葡萄球菌、乳杆菌、肠杆菌、双歧杆菌数量与血清hsCRP水平无显著相关性(均P>0.05)。结论H.pylori根除治疗能有效改善慢性胃炎患者临床症状,提高H.pylori根除率,降低血清hsCRP水平,但会造成患者肠道菌群失调,且肠道菌群数量与血清hsCRP水平无显著相关性。     

血清尿酸、超敏C 反应蛋白与老年永久性房颤的关系

目的:探讨血清尿酸(UA)、超敏C反应蛋白(hs-CRP)、左心房内径(LAD)与老年永久性房颤的相关性。方法:选取2012年8月至2015年9月于首都医科大学宣武医院综合科住院的老年永久性房颤患者78例作为房颤组,选取同期住院窦性心律且无房颤病史的老年患者72例作为对照组,记录各组一般资料及超声心动图、尿酸、hs-CRP水平等,进行比较及相关性分析。结果:1与对照组相比,房颤组血清UA及hs CRP水平、左心房内径大小明显增高(P0.05),2以房颤是否发生为因变量,多因素logistic回归分析显示血清尿酸(OR=2.016,P=0.004)、左房内径(OR=4.180,P=0.001)和hs-CRP(OR=2.846,P=0.002)是老年永久性房颤发生的独立危险因素。3房颤组患者血清尿酸与hs-CRP水平呈正相关(r=0.28,P0.05),尿酸与左心房内径大小呈轻度正相关(r=0.12,P0.05)。结论:血清尿酸、hs-CRP、左房内径是老年永久性房颤发生的独立危险因素;尿酸参与的炎症反应和结构重构可能参与老年永久性房颤的发生。     

沙丁胺醇联合福多司坦治疗慢性阻塞性肺疾病稳定期的疗效及对患者血清IL-6、TNF-α、hs-CRP水平的影响

目的:探讨沙丁胺醇联合福多司坦治疗慢性阻塞性肺疾病稳定期的临床疗效及对患者血清白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)、超敏c反应蛋白(hs-CRP)水平的影响。方法:选择2016年1月到2017年1月我院接诊的稳定期慢性阻塞性肺病患者100例作为研究对象,按照随机数表法分为观察组(n=51)和对照组(n=49)。对照组使用沙丁胺醇治疗,观察组采用沙丁胺醇联合福多司坦治疗。比较两组治疗后的疗效、治疗前后血清IL-6、TNF-α、hs-CRP、肺功能的变化及不良反应的发生情况。结果:治疗后,观察组临床疗效总有效率(94.12%)显著高于对照组(75.51%,P0.05)。两组患者治疗后血清IL-6、TNF-α、hs-CRP水平均治疗前均明显下降,且观察组患者血清IL-6、TNF-α、hs-CRP水平均明显低于对照组(P0.05);两组治疗后各第1秒用力呼气容积(FEV1)、用力肺活量(FVC)、最大呼气流量(PEF)较治疗前均显著升高(P0.05),且观察组FEV1、FVC、PEF均明显高于对照组(P0.05);两组患者不良反应总发生率分别19.61%、38.78%,观察组显著低于对照组(P0.05)。结论:沙丁胺醇联合福多司坦治疗慢性阻塞性肺疾病稳定期的临床疗效和安全性均显著优于单用沙丁胺醇治疗,可能与其有效改善患者血清IL-6、TNF-α、hs-CRP水平有关。     

超声对早期糖尿病肾病的诊断价值及肾动脉血流阻力指数与血清hs-CRP、VEGF的关系分析

目的:研究彩色多普勒超声对早期糖尿病肾病的诊断价值及肾动脉血流阻力指数与血清超敏C反应蛋白(hs-CRP)、血管内皮生长因子(VEGF)的关系。方法:选取从2017年2月～2018年2月兰州大学第二医院收治的早期糖尿病肾病患者50例记为病变组,另取同期于该院进行体检的健康人员50例记为对照组。分别对两组人员进行彩色多普勒超声检查,比较肾血流参数。采用酶联免疫吸附法检测两组人员血清hs-CRP、VEGF水平,并作指标间的相关性分析。结果:病变组肾主动脉、肾锥体两侧叶间动脉、肾段动脉的收缩期峰值速度、舒张期最低速度较对照组降低,病变组肾主动脉、肾锥体两侧叶间动脉、肾段动脉的阻力指数较对照组升高(均P0.05)。病变组血清hs-CRP、VEGF水平较对照组升高(均P0.05)。经Pearson相关性分析显示:早期糖尿病肾病患者肾主动脉、肾锥体两侧叶间动脉、肾段动脉的血流阻力指数与血清hs-CRP、VEGF均呈正相关关系(均P0.05)。结论:彩色多普勒超声应用于早期糖尿病肾病的诊断价值较高,且肾动脉血流阻力指数与血清hs-CRP、VEGF密切相关,临床工作中通过联合检测血清hs-CRP、VEGF,从而有助于早期糖尿病肾病的诊断。     

Implication of a pepper h-type thioredoxin in type I- and II-nonhost resistance to Xanthomonas axonopodis

Thioredoxins (TRXs) are distributed ubiquitously in prokaryotic and eukaryotic organisms. Plants have the most complex forms of TRXs. The functional roles of such TRXs have been studied in abiotic stress but their roles in plant defense responses against biotic stresses have been less well studied. Here, we identified an h-type TRX gene from pepper, CaTRXh1, and characterized its possible effect on Type II nonhost resistance, which entails localized programmed cell death in response to nonhost pathogens. Peptide sequences of CaTRXh1 showed a high degree of similarity with TRXhs from tobacco and Arabidopsis thaliana. Southern blot analyses revealed that CaTRXh1 was present as a single copy in the pepper genome. Intriguingly, leaf infiltration by Xanthomonas axonopodis pv. glycines 8ra, eliciting a visible type II nonhost hypersensitive response (HR), and its type III secretion-system null mutant 8–13, eliciting a type I nonhost non-HR, both induced CaTRXh1 at a level similar to that of pathogenesis-related protein 4, an HR marker gene in pepper. More surprisingly, expression of CaTRXh1 was significantly increased when X. axonopodis pv. vesicatoria race 3 infiltrated the leaf of a pepper cultivar containing a resistance gene, but not with infiltration of a susceptible pepper cultivar. Taken together, our study suggests that the expression of CaTRXh1 has a critical role in HR-mediated active defense responses in pepper. GenBank accession number: EF371503.     

Molecular and biochemical characterization of the<Emphasis Type="Italic"> Capsicum annuum calcium-dependent protein kinase 3</Emphasis> (<Emphasis Type="Italic">CaCDPK3</Emphasis>) gene induced by abiotic and biotic stresses

The isolated full-length Capsicum annuum calcium-dependent protein kinase 3 (CaCDPK3) cDNA clone was selected from the chili pepper expressed sequence tag database (). Phylogenetic analysis based on the deduced amino acid sequence of CaCDPK3 cDNA revealed significant sequence similarity to the winter squash (Cucurbita maxima) CmCPK2 gene (81% identity). Genomic gel blot analysis disclosed that CaCDPK3 belongs to a multigene family in the pepper genome. CaCDPK3 expression was root tissue-specific, as shown by Northern blot data. The gene was rapidly induced in response to various osmotic stress factors and exogenous abscisic acid application in pepper leaves. Moreover, CaCDPK3 RNA expression was induced by an incompatible pathogen and by plant defense-related chemicals such as ethephon, salicylic acid and jasmonic acid. The biochemical properties of CaCDPK3 were investigated using a CaCDPK3 and glutathione S-transferase (GST) fusion protein. The recombinant proteins retained calcium-binding ability, and displayed autophosphorylation activity in vitro in a calcium-dependent manner. Further transient-expression studies showed that CaCDPK3 fused with soluble modified green fluorescent protein (smGFP) localized to the cytosol in chili pepper protoplasts. We propose that CaCDPK3 is implicated in biotic and abiotic stresses in pepper plants.