Split Nano luciferase complementation for probing protein‐protein interactions in plant cells

Deciphering protein‐protein interactions (PPIs) is fundamental for understanding signal transduction pathways in plants. The split firefly luciferase (Fluc) complementation (SLC) assay has been widely used for analyzing PPIs. However, concern has risen about the bulky halves of Fluc interfering with the functions of their fusion partners. Nano luciferase (Nluc) is the smallest substitute for Fluc with improved stability and luminescence. Here, we developed a dual‐use system enabling the detection of PPIs through the Nluc‐based SLC and co‐immunoprecipitation assays. This was realized by coexpression of two proteins under investigation in fusion with the HA‐ or FLAG‐tagged Nluc halves, respectively. We validated the robustness of this system by reproducing multiple previously documented PPIs in protoplasts or Agrobacterium‐transformed plants. We next applied this system to evaluate the homodimerization of Arabidopsis CERK1, a coreceptor of fungal elicitor chitin, and its heterodimerization with other homologs in the absence or presence of chitin. Moreover, split fragments of Nluc were fused to two cytosolic ends of Arabidopsis calcium channels CNGC2 and CNGC4 to help sense the allosteric change induced by the bacterial elicitor flg22. Collectively, these results demonstrate the usefulness of the Nluc‐based SLC assay for probing constitutive or inducible PPIs and protein allostery in plant cells.     

喉罩通气下七氟醚全凭吸入麻醉在小儿先天性心脏病介入手术的临床麻醉效果

目的:探讨喉罩通气下七氟醚全凭吸入麻醉在小儿先天性心脏病介入手术的临床麻醉效果。方法:选取2017年4月～2019年5月期间我院收治的行先天性心脏病介入手术患儿98例,根据随机数字表法将其分为对照组(n=49)和研究组(n=49)。对照组给予氯胺酮诱导,全凭丙泊酚维持,面罩吸氧;研究组给予全凭七氟醚诱导、维持,喉罩通气。比较两组患儿麻醉前(T0)、切皮前(T1)、切皮后1 min(T2)、切皮后30 min(T3)、术后(T4)的血流动力学指标[平均动脉压(MAP)、心率(HR)]及应激反应指标[血糖、皮质醇],记录两组患儿手术时间、麻醉诱导时间、术后苏醒时间等围术期指标情况。记录两组围术期不良反应发生情况。结果:研究组手术时间、麻醉诱导时间、术后苏醒时间均短于对照组(P0.05)。两组T0时间点血糖、MAP、皮质醇、HR比较差异无统计学意义(P0.05);对照组T1～T4时间点MAP、血糖、皮质醇、HR均较T0升高(P0.05);研究组T1～T4时间点血糖、MAP、皮质醇、HR与T0时间点比较无差异(P0.05);研究组T1～T4时间点血糖、MAP、皮质醇、HR低于对照组(P0.05)。两组不良反应发生率比较无差异(P0.05)。结论:小儿先天性心脏病介入手术中应用喉罩通气下七氟醚全凭吸入麻醉,诱导迅速且安全、术后苏醒快、手术时间短,可有效维持血流动力学稳定,减少应激反应。     

微创肺表面活性物质对呼吸窘迫综合征新生儿肝肾功能、氧合功能及呼吸功能的影响

目的:探讨微创肺表面活性物质(pulmonary surfactant,PS)对呼吸窘迫综合征新生儿氧合功能、肝肾功能及呼吸功能的影响。方法:选择2018年3月至2019年3月我院收治的66例新生呼吸窘迫综合征的患儿作为研究对象,并按照随机数字表法分为观察组(n=33)和对照组(n=33)。对照组患儿采取常规的治疗方式,观察组患儿则在对照组治疗基础上应用微创PS治疗。观察比较两组患儿的动脉血气指标、Ⅱ型肺泡细胞表面抗原(KL-6)、巨噬细胞移动抑制因子-1(MIF-1)、高迁移率族蛋白1(HMGB-1)、肝肾功能、氧合指数及呼吸机参数。结果:治疗后,两组的氧分压(PaO_2)、二氧化碳总量(TCO_2)、氧饱和度(SaO_2)均较治疗前显著增高;且观察组以上指标均高于对照组。两组的Ⅱ型肺泡细胞表面抗原(KL-6)、巨噬细胞移动抑制因子-1 (MIF-1)、高迁移率族蛋白1(HMGB-1)均较治疗前显著降低,且观察组的以上指标均低于对照组。两组的谷草转氨酶(AST)、谷丙转氨酶(ALT)、尿素氮(BUN)、肌酐(CRE)水平均较治疗前显著降低,且观察组的以上指标均明显低于对照组。观察两组的呼吸机参数和氧合指数,发现两组的吸入氧浓度(FiO_2)、吸气峰压(PIP)、呼吸末正压(PEEP)、氧合指数(PaO_2/FiO_2,OI)均较治疗前有所改善,且观察组的以上指标均要优于对照组(P0.05)。结论:应用微创PS治疗新生儿呼吸窘迫综合征的效果显著,能明显改善患儿的动脉血气指标、肝肾功能以及氧合功能,减轻炎症反应,并减少机械通气的参数。     

西格列汀通过增加肾组织中SOCS 1和Podocalyxin的表达改善糖尿病肾病大鼠肾功能

目的:探讨西格列汀对糖尿病肾病大鼠肾功能及肾组织中细胞因子信号传导负调控因子1(Suppressors of cytockine signaling,SOCS 1)和足细胞特异蛋白抗体(Podocalyxin)表达的影响。方法:将大鼠随机分为4组:对照组,模型组,西格列汀组和贝那普利组。模型组、西格列汀组和贝那普利组采用腹腔注射链脲佐菌素建立模型,对照组给予腹腔注射等量生理盐水。造模成功后,西格列汀组(n=8)和贝那普利组(n=8)分别灌胃给予7 mg/kg/d的西格列汀和贝那普利。模型组(n=8)和对照组(n=10)均给予等体积的蒸馏水灌胃,连续8周。检测并对比各组大鼠代谢相关指标,肾组织纤维化程度指标,肾组织中炎症因子水平以及Podocalyxin、SOCS 1和结蛋白(Desmin)表达。结果:干预8周后,与对照组对比,模型组空腹血糖、糖化血红蛋白、甘油三酯、总胆固醇、24 h尿蛋白排泄率、肌酐、体重、肾组织转化生长因子-β1(Transforming growth factor,TGF-β1)、白介素(Interleukin,IL)-6、IL-1β、SOCS 1和Desmin水平均显著增加,Ⅳ型胶原蛋白(Collagen-Ⅳ,C-Ⅳ)、纤维连接蛋白(Fibronectin,FN)、层黏连蛋白(Laminin,LN)和Podocalyxin水平显著降低(P0.05);与模型组对比,西格列汀组和厄贝沙坦组空腹血糖、糖化血红蛋白、甘油三酯、总胆固醇、24 h尿蛋白排泄率、肌酐、体重、肾组织TGF-β1、IL-6、IL-1β和Desmin水平均显著降低,Podocalyxin和SOCS 1蛋白表达增加(P0.05)。但厄贝沙坦组与西格列汀组以上各指标对比差异无统计学意义(P0.05)。结论:西格列汀可能通过增加Podocalyxin和SOCS 1蛋白表达,降低肾组织中炎症因子和Desmin蛋白表达,进而改善糖尿病肾病大鼠肾纤维化和肾功能。     

复方α-酮酸联合血液透析和血液灌流治疗慢性肾衰竭的疗效及对钙磷代谢的影响

目的:研究复方α-酮酸联合血液透析和血液灌流治疗慢性肾衰竭(CRF)的临床疗效及对钙磷代谢的影响。方法:选择94例CRF患者为研究对象,按照随机数表法将患者分为联合组与对照组各47例。对照组采用血液透析、血液灌流进行治疗,联合组则在对照组基础之上联合使用复方α-酮酸治疗。比较治疗前及治疗6个月后两组患者肾功能指标[血清胱抑素C(Cys C)、血清肌酐(Scr)、血尿素氮(BUN)]及钙磷代谢(血钙、血磷)变化,并分析治疗6个月后两组患者疗效及治疗6个月内药物不良反应发生情况差异。结果:治疗6个月后,联合组治疗总有效率明显高于对照组(P0.05);两组患者Cys C、Scr、BUN及血磷水平均较治疗前显著降低,且联合组明显低于同一时间对照组(P0.05)。两组患者血钙水平均较治疗前显著升高,且联合组明显高于同一时间对照组(P0.05)。治疗6个月内,两组患者药物不良反应总发生率比较差异均无统计学意义(P0.05)。结论:复方α-酮酸联合血液透析和血液灌流治疗CRF的疗效显著,且能够改善肾功能与钙磷代谢,对患者疾病转归有利。     

Molecular phylogeny and species delimitation of Stachyuraceae: Advocating a herbarium specimen‐based phylogenomic approach in resolving species boundaries

Species concept and delimitation are fundamental to taxonomic and evolutionary studies. Both inadequate informative sites in the molecular data and limited taxon sampling have often led to poor phylogenetic resolution and incorrect species delineation. Recently, the whole chloroplast genome sequences from extensive herbarium specimen samples have been shown to be effective to amend the problem. Stachyuraceae are a small family consisting of only one genus Stachyurus of six to 16 species. However, species delimitation in Stachyurus has been highly controversial because of few and generally unstable morphological characters used for classification. In this study, we sampled 69 individuals of seven species (each with at least three individuals) covering the entire taxonomic diversity, geographic range, and morphological variation of Stachyurus from herbarium specimens for genome‐wide plastid gene sequencing to address species delineation in the genus. We obtained high‐quality DNAs from specimens using a recently developed DNA reconstruction technique. We first assembled four whole chloroplast genome sequences. Based on the chloroplast genome and one nuclear ribosomal DNA sequence of Stachyurus, we designed primers for multiplex polymerase chain reaction and high throughput sequencing of 44 plastid loci for species of Stachyurus. Data of these chloroplast DNA and nuclear ribosomal DNA internal transcribed spacer sequences were used for phylogenetic analyses. The phylogenetic results showed that the Japanese species Stachyurus praecox Siebold & Zucc. was sister to the rest in mainland China, which indicated a typical Sino‐Japanese distribution pattern. Based on diagnostic morphological characters, distinct distributional range, and monophyly of each clade, we redefined seven species for Stachyurus following an integrative species concept, and revised the taxonomy of the family based on previous reports and specimens, in particular the type specimens. Furthermore, our divergence time estimation results suggested that Stachyuraceae split from its sister group Crossosomataceae from the New World at ca. 54.29 Mya, but extant species of Stachyuraceae started their diversification only recently at ca. 6.85 Mya. Diversification time of Stachyurus in mainland China was estimated to be ca. 4.45 Mya. This research has provided an example of using the herbarium specimen‐based phylogenomic approach in resolving species boundaries in a taxonomically difficult genus.     

不同分级及转归脓毒症患者血清PCT、D-D、CRP及血小板相关参数检测的临床意义

目的:探讨不同分级及转归脓毒症患者的血清降钙素原(PCT)、D-二聚体(D-D)、C反应蛋白(CRP)及血小板相关参数检测的临床意义。方法:回顾性分析2015年3月至2018年8月期间中国人民解放军西部战区总医院收治的92例脓毒症患者的临床资料,分析不同分级及转归脓毒症患者的血清中PCT、D-D和CRP水平、急性病生理与慢性健康评价系统Ⅱ(APACHEⅡ)评分及血小板相关参数[血小板计数(PLT)、血小板平均容积(MPV)、血小板分布宽度(PDW)、大型血小板比率(P-LCR)],并分析脓毒症患者PCT、D-D、CRP水平以及血小板相关参数与APACHEⅡ评分的相关性。结果:全身炎症反应综合征组、轻度脓毒症组、严重脓毒症组、脓毒性休克组血清PCT、D-D和CRP水平、MPV、PDW、P-LCR、APACHEⅡ评分逐渐升高(P0.05),PLT逐渐降低(P0.05)。存活组患者血清PCT、D-D、CRP水平、MPV、PDW、P-LCR、APACHEⅡ评分均低于死亡组(P0.05),PLT高于死亡组(P0.05)。Pearson相关分析显示,脓毒症患者血清PCT、D-D、CRP、MPV、PDW、P-LCR与APACHE II评分呈正相关(P0.05),PLT与APACHE II评分呈负相关(P0.05)。结论:脓毒症患者血清PCT、D-D、CRP及血小板相关指标可能参与了脓毒症的发展,通过检测其血清PCT、D-D、CRP水平及血小板相关参数可评估脓毒症患者的病情和预后。     

加味星蒌承气汤对急性缺血性脑卒中患者神经功能及血脂、血液流变学的影响

目的:探讨加味星蒌承气汤对急性缺血性脑卒中患者神经功能及血脂、血液流变学的影响。方法:选取2017年8月～2019年6月期间我院收治的急性缺血性脑卒中患者96例,将入选患者根据随机数字表法分为对照组(n=48)和研究组(n=48),对照组患者予以常规西医治疗,研究组患者在对照组基础上联合加味星蒌承气汤治疗,对比两组患者疗效、神经功能及血脂、血液流变学情况,记录两组患者治疗期间不良反应情况。结果:研究组治疗8 d后的临床总有效率为91.67%(44/48),显著高于对照组患者的72.92%(35/48)(P0.05)。两组治疗8 d后加拿大神经功能评分量表(CNS)、美国国立卫生研究所卒中量表(NIHSS)评分均下降(P0.05),且研究组低于对照组(P0.05)。两组治疗8 d后总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG)、全血黏度高切、全血黏度低切、血浆黏度、纤维蛋白原均下降(P0.05),且研究组低于对照组(P0.05);高密度脂蛋白胆固醇(HDL-C)升高(P0.05),且研究组高于对照组(P0.05)。两组不良反应发生率比较无明显差异(P0.05)。结论:急性缺血性脑卒中患者采用加味星蒌承气汤治疗,疗效显著,可有效改善患者神经功能及血脂、血液流变学,且安全性较好。     

Epidemiological,Clinical and Serological Characteristics of Children with Coronavirus Disease 2019 in Wuhan: A Single-centered,Retrospective Study

In December 2019, SARS-CoV-2 was first detected in the samples obtained from three adult patients who suffered from an unknown viral pneumonia in Wuhan (Li et al. 2020). This unknown viral pneumonia is further named as coronavirus disease 2019 (COVID-19) by the World Health Organization. To date, the number of new COVID-19 cases has continued to skyrocket and the impact of SARS-CoV-2 on humans is far greater than any pathogen of this century in both breadth and depth. Previous studies have shown that adults with COVID-19 have symptoms of fever, dry cough, dyspnea, fatigue and lymphocytopenia. Moreover, COVID-19 is more likely to cause death in the elderly, especially those with chronic comorbidities (Huang et al. 2020). In Wuhan, more than 50, 000 COVID-19 cases have been confirmed, including over 780 pediatric patients, and only one child death case (Lu et al. 2020). Although the number of children cases was far fewer than that of adults, COVID-19 might endanger children's health and the information on children remains limited, especially in serological study. In the retrospective study, the investigators analyzed the epidemiological, clinical and serological characteristics of children with COVID-19 in Wuhan in the early stages of the outbreak, which might provide theoretical and practical help in controlling COVID-19 and similar emerging infectious diseases in the future.