为了探究NFIB(nuclear factor I/B)基因在膀胱癌中的生物学功能,该研究应用CRISPR-Cas9技术构建NFIB敲除的膀胱癌细胞株,然后通过转录组学技术分析对照细胞NFIB-NC与NFIB敲除细胞株NFIB-KO-Ⅰ和NFIB-KO-Ⅱ之间的差异表达基因,同时对其进行KEGG富集分析,并通过Western blot和qRT-PCR技术对转录组学结果进行验证.结果显示,与对照细胞NFIB-NC相比,NFIB敲除细胞株NFIB-KO-Ⅰ中有134个差异表达基因,其中上调基因62个,下调基因72个;NFIB-KO-Ⅱ中有131个差异表达基因,表达上调和下调的基因分别为50个和81个.KEGG分析结果显示,差异表达基因与PI3K-AKT信号通路密切相关,Western blot结果证实敲除NFIB基因后,膀胱癌细胞中的p-AKT水平显著上调.qRT-PCR结果表明,敲除NFIB后,ITGA4基因表达水平上调,TNC和ANGPT4基因表达水平下调.该研究初步揭示了 NFIB基因在膀胱癌细胞中调控的分子信号通路,也为后续研究NFIB基因介导膀胱癌发生发展的分子机制提供了依据.

Objective:Based on the high-throughput detection technique of multiplex PCR combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, constructing the characteristic SNP profiles of different strains, and establishing a rapid, accurate and highly sensitive method for the diagnosis of bloodstream infection pathogens.Methods:Seven kinds of pathogens such as common Escherichia coli were selected as target. The multiple PCR reaction conditions was optimized, and the characteristic peaks of each target bacteria were detected by MALDI-TOF MS to establish the joint detect system. Common primer pairs and central homo-sequence primer pairs were designed to analyse the formation of primer dimer. Using simulated bacterial infection blood samples with detection system to determine specificity and sensitivity. One hundred and fifty blood samples from suspected bacteremia patients were collected from June to September 2020 in a hospital in Beijing, and the identification results were compared to traditional identification method of clinical application that are using χ 2 test. Results:The cycle threshold (Ct) value of the central homo-sequence primers that were designed were more than 38, with a delay of 6-10 cycles. The joint mass spectrometry detection system could detect seven kinds of bacteria divided into two groups at the same time. The target bacteria can be detected specific product of the peak, and the clinical strains other than the target strains only had primer peaks. All maps had non-specific miscellaneous peaks. The sensitivity of Escherichia coli could reach 50 CFU/ml, and the detection limit of other bacteria was 100 CFU/ml. The detection results of 150 patients showed that 46 cases were positive by traditional method. The positive rate was 30.67% (46/150), including two cases of mixed infection. Forty-eight cases were positive by mass spectrometry, and the positive rate was 32.0% (48/150), including three cases of mixed infections. The negative coincidence rate was 100% (101/101). The comparison of the two methods showed that the P=0.625>0.01, the Kappa=0.938, the sensitivity and specificity was 97.82%(45/46) and 97.11%(101/104), respectively. There was no significant difference between the two methods, and the results of nucleic acid mass spectrometry could also be used in clinic. Conclusions:The established detection system can not only quickly and accurately detect seven common pathogens causing bloodstream infection, and effectively shorten the time needed for traditional culture and identification, but also can detect multiple bacterial mixed infections at the same time to make up for the possibility of missed detection. Besides, the method can also be used to identify other bacteria.

目的 原核表达、纯化结核分枝杆菌(Mycobacterium tuberculosis,Mtb)早期分泌蛋白MPT64,并初步评估其在结核病(tuberculosis,TB)血清学诊断中的价值.方法 PCR扩增Mtb标准菌株H37Rv mpt64基因,克隆至原核表达载体pET28a(+),构建重组质粒pET28a-mpt64,转化大肠埃希菌BL21(DE3)后,进行重组蛋白的原核诱导表达及纯化.将600 μg纯化的MPT64蛋白经皮下多点免疫新西兰大白兔,隔周免疫1次,共4次,末次免疫2周后,耳缘静脉采血分离血清,ELISA法检测血清效价,Western blot法鉴定抗原特异性.收集44例TB确诊患者(TB组)和36名健康对照人群[HC(heahhy control)组]血清,间接ELISA法检测血清中MPT64抗原的特异性IgG、IgA抗体水平,抗原对照为GroES和Ag85A.结果 重组表达质粒pET28a-mpt64经双酶切鉴定及测序证明构建正确;表达的重组MPT64蛋白主要以包涵体形式存在,部分以可溶性形式存在,经Ni2+-NTA亲和纯化后,纯度可达90％,蛋白浓度约2.0 mg/mL;MPT64兔多克隆抗血清效价达1:2048000,纯化的MPT64蛋白可与其发生反应,在相对分子质量约24000处可见特异性蛋白条带;TB组血清中MPT64抗原特异性IgG和IgA水平均显著高于HC组(P＜0.05),而GroES、Ag85A抗原特异性IgA和IgG水平差异无统计学意义(P＞0.05).结论 重组MPT64蛋白具有较好的血清学诊断效能,可作为TB血清学早期诊断的候选抗原之一.

目的 建立应用血栓弹力图(TEG)及凝血指标综合评估上消化道出血患者凝血功能的输血结局预测模型.方法 从输血(科)管理系统和医院信息系统(HIS)系统收集浙江省人民医院及其淳安分院消化内科2018年6月～2021年6月收治的101名符合临床诊断标准的上消化道出血患者,根据结局是否输血分为输血组(n=56)和未输血组(n=45),以及按照肝硬化与否分为肝硬化组(n=74)和非肝硬化组(n=27),同时收集40名非上消化道出血患者的阴性对照组.对比各组的TEG检测R、K、α、MA参数,凝血功能检测PT、INR、APTT、TT、Fib,血常规检测Hb、Plt、WBC、NEUT％,以及生化检测Alb、SCr、ALT、AST、GGT等指标(值);分析TEG指标与传统凝血功能指标的相关性;采用单因素和多因素分析,筛选输血相关因素建立预测模型.结果 输血组与未输血组比较:TEG的K(min)为3.86±3.12 vs 2.50±1.47,α(°)为54.00±14.08 vs 61.05±10.88,MA(mm)为51.12±13.37 vs 58.26±11.08(P＜0.01);凝血指标检测PT(s)为16.36±7.45 vs 13.44±1.50,Fib(g)为1.59±0.87 vs 2.35±1.09(P＜0.01);血常规检测NEUT％为0.75±0.13 vs 0.66±0.15,Hb(g/L)为68.04±14.49 vs 100.73±22.92(P＜ 0.01);生化检测Alb (g/L)为29.73±6.08 vs 33.73±7.19,SCr(nmol/L)为99.50±53.55 vs 76.25±19.28 (P＜0.01).相关性分析:APTT与R、K值呈正相关,与α、MA值呈负相关;Fib与K值呈负相关,与α、MA值呈正相关;Plt与K值呈负相关,与α[、MA值呈正相关(P＜0.01).单因素分析:将得到8个输血前因素K、MA、PT、Fib、NEUT％、Hb、Alb、SCr做Logistic回归,建立输血预测模型,ROC曲线最佳输血阈值(患者输血预测值)为0.448,灵敏度92.9％,特异度88.9％,AUC0.969.结论 综合上消化道出血患者TEG、凝血功能、血常规和生化等检测指标,建立Logistic回归模型对预测患者输血结局有明显的关联性,有较好的临床实用性.

目的 探讨一例Krabbe病患者的遗传病因,并对家系成员进行基因突变分析及产前诊断.方法 应用全外显子测序技术对Krabbe病患者进行致病突变筛查,结合临床表型,确定候选基因的致病位点,Sanger测序验证夫妻双方GALC基因,孕妇进行绒毛穿刺和羊水穿刺,对GALC基因测序并进行产前诊断.结果 全外显子测序结果显示家系中Krabbe病患者存在GALC基因c.599C＞A (p.Ser200*)和c.461C＞A (p.Pro154His)位点复合杂合变异,其父亲为c.461C＞A(p.Pro154His)位点杂合变异携带者,母亲为c.599C＞A (p.Ser200*)杂合变异携带者.绒毛和羊水检测GALC基因为c.461C＞A (p.Pro154His)位点杂合变异.结论 GALC基因c.599C＞A (p.Ser200*)和c.461C＞A (p.Pro154His)位点复合杂合变异是该家系中Krabbe病患者的发病原因,胎儿绒毛和羊水检测结果与先证者不同,为该家系遗传咨询和产前诊断提供有力证据,有效预防出生缺陷.

目的 探讨基于特异常规引物(即引物不需折叠成特定二级结构)靶向恒温扩增重复DNA序列的方法,并测试其检测结核分枝杆菌(Mycobacterium tuberculosis,Mtb)的可能性.方法 以合成的重复DNA或以抽提的细菌基因组DNA为模板,用Bst 2.0 WarmStart DNA聚合酶进行恒温扩增.用琼脂糖凝胶电泳检测扩增结果.结果 合成的重复DNA可用其特异常规引物进行恒温扩增.进一步,Mtb H37RvⅡ型分枝杆菌散在重复单位(mycobacterial interspersed repetitive units,MIRUs)可用其特异常规引物对(即ⅡMIRU-F和Ⅱ_MIRU-R)或单引物(即Ⅱ_MIRU-F)进行恒温扩增.相比于非分枝杆菌菌株、非结核分枝杆菌菌株、Mtb复合物菌株和临床分离株,ⅡMIRU-F能够高特异地扩增Mtb菌株.Ⅱ_MIRU-F针对Mtb H37Rv基因组DNA的检测下限较高,且不能特异地区分Mtb阴性痰液样本和Mtb阳性痰液样本.结论 常规引物可恒温扩增重复DNA序列(包括MtbⅡ型MIRUs);MtbⅡ型MIRUs特异引物Ⅱ_MIRU-F不适合用于痰液样本的检测.

该文通过shRNA干扰技术敲低IscU2干扰细胞IscU2的表达,研究了干扰IscU2对非小细胞肺癌(NSCLC)细胞NCI-H520增殖、迁移及侵袭能力的影响.构建了稳定低表达IscU2的非小细胞肺癌细胞系NCI-H520;采用CCK-8和平板克隆实验检测细胞的增殖能力;流式细胞仪检测细胞周期、凋亡、ROS、线粒体膜电位变化情况;Transwell实验检测细胞迁移及侵袭能力;Westernblot检测相关蛋白的表达.结果 表明,干扰IscU2后,非小细胞肺癌细胞的增殖及克隆形成能力降低;细胞周期停滞在G1/G0期,同时伴随有p-AKT和Cyclin D1蛋白含量的下降;细胞晚期凋亡率明显增加,凋亡蛋白Cleaved-caspase3和Cleaved-PARP表达上调;细胞迁移和侵袭能力降低,上皮标志物E-Cadherin表达上调,间质标志物N-Cadherin和Snail表达下调;细胞ROS积累和线粒体膜电位下降.该研究结果表明,干扰IscU2显著抑制非小细胞肺癌的增殖、迁移、侵袭能力和上皮-间质转化,这为非小细胞肺癌的诊断和治疗提供了新的潜在靶点和视角.

YAP(yes-associated protein)是Hippo信号通路中发挥转录共激活作用的蛋白.已知YAP能够参与细胞的多个代谢过程,但YAP是否参与了线粒体功能的调控尚不清楚.该研究发现,无论是化合物抑制YAP功能还是基因敲低YAP表达水平均能够显著提升线粒体呼吸链组装水平,并促进线粒体呼吸能力的上升和膜电位的升高.初步的机制分析表明,YAP基因功能的抑制可正向调控促进线粒体生物能学的相关转录因子Nrf1、RXRα和POLG的表达;负向调控抑制线粒体生物能学的转录因子HIF1-alpha的表达.进一步的生物学功能分析表明,葡萄糖应激或者葡萄糖剥夺下线粒体功能的上升部分依赖于YAP表达量的抑制.综上,该研究发现,YAP可通过调节线粒体功能调控相关转录因子的表达来影响线粒体功能,且葡萄糖应激条件下线粒体功能的维持可通过YAP途径实现.

目的:探讨高迁移率族蛋白B1(HMGB1)在结肠癌细胞中的表达和对结肠癌细胞增殖、迁移和侵袭的影响.方法:采用qPCR与Western blot法检测人结肠癌SW620细胞与正常结肠上皮细胞FHC中HMGB1的mRNA和蛋白表达.通过Lipofectamine 3000转染质粒构建稳定下调HMGB1表达(shHMGB1)的SW620细胞,同时设置阴性对照(shNC)细胞和空白对照(blank)细胞;采用CCK-8、平板集落形成实验和Transwell实验检测细胞的增殖、迁移及侵袭能力.采用Western blot检测HMGB1表达下调对p-ERK、ERK、c-Myc、基质金属蛋白酶(MMP)-2/9、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、Bcl-2及Bax的蛋白表达水平的影响.结果:SW620细胞中HMGB1在mRNA及蛋白水平表达均高于正常结肠上皮细胞FHC(P<0.05).经qPCR与Western blot验证,HMGB1低表达细胞构建成功.与blank组和shNC组相比,shHMGB1组细胞增殖、迁移和侵袭能力受到显著抑制(P<0.01).Western blot结果表明,同blank组和shNC组比较,shHMGB1组细胞的MMP-2、MMP-9、N-cadherin、c-Myc、Bcl-2及ERK磷酸化蛋白表达显著下调,Bax蛋白表达显著上调(P<0.05).结论:HMGB1可促进结肠癌SW620细胞的增殖、迁移和侵袭能力,ERK/c-Myc信号通路参与了这一过程.

为构建一种能够高效、同时吸附水中多种重金属离子的大肠杆菌,利用融合蛋白表达技术,首先将大肠杆菌前脂蛋白信号肽Lpp、膜蛋白OmpA的N端部分氨基酸和铁硫簇组装蛋白IscA的编码基因序列进行融合,构建pET-Lpp-OmpA-IscA表达载体,将此载体导入大肠杆菌BL21菌株.在IPTG诱导下,IscA蛋白可表达于细胞膜表面.然后对IscA膜表面表达菌株对重金属的吸附能力进行评估,包括测定最大吸附容量、绘制吸附浓度依赖曲线和时间依赖曲线,以及对菌株清除工业污水中重金属的性能进行初步探索.研究结果表明,与本底对照菌株相比,IscA蛋白在细胞膜表面表达能够使菌株对水中的Cu2+、Ni2+、Cd2+、pb2+、As3+、Co2+、Hg2+这7种重金属的吸附能力提高2～5倍不等,并且在pH为6～8范围内保持其吸附能力基本不变.此菌株能够在30 min内将各种重金属溶液中超标5倍的金属含量降低至最大允许排放浓度以下,并且对吸附的重金属具有不同程度的回收能力和菌株再生能力.此外,该菌株能够同时吸附工业污水中的多种重金属,有效降低各种重金属含量.因此,利用膜表面表达技术对大肠杆菌进行改造,成功提高了大肠杆菌对多种重金属的吸附能力,为利用微生物治理环境重金属污染提供了良好的应用前景.

目的:研究二甲双胍是否通过下调c-Myc增强人乳腺癌MDA-MB-231细胞对他莫昔芬的敏感性.方法:CCK-8法、平板集落形成实验、流式细胞术和Transwell实验检测二甲双胍和他莫替芬单独/联合作用对MDA-MB-231细胞活力、集落形成能力、肿瘤细胞凋亡率及迁移和侵袭能力的影响;Western blot和免疫组化实验检测细胞和肿瘤组织中c-Myc的表达水平;建立SCID小鼠MDA-MB-231细胞皮下移植瘤模型,以研究二甲双胍和他莫昔芬在体内对三阴性乳腺癌的影响.结果:与对照组和单独用药组相比,二甲双胍联合他莫昔芬协同抑制MDA-MB-231细胞的活力和迁移能力,对集落形成、凋亡诱导和侵袭能力表现为相加效应(P<0.01);二甲双胍和他莫昔芬单独或者联合下调c-Myc蛋白的水平(P<0.01).进一步研究表明,二甲双胍和他莫昔芬可能通过下调c-Myc抑制SCID小鼠皮下移植瘤的生长(P<0.01).结论:二甲双胍联合他莫昔芬通过下调c-Myc抑制三阴性乳腺癌MDA-MB-231细胞的增殖、迁移和侵袭以及SCID小鼠的肿瘤生长.二甲双胍联合他莫昔芬有望成为三阴性乳腺癌的一种有效的治疗手段.

KCTD5是钾离子通道四聚体结构域(KCTD)蛋白家族的一员.KCTD5可作为Cullin3的接头蛋白发挥生物学功能,但是其与急性髓系白血病(acute myeloid leukemia,AML)的关系尚不清楚.通过对癌症与肿瘤基因图谱(the Cancer of Genome Atlas,TCGA)数据库的分析,发现KCTD5 mRNA表达水平与AML病人生存期呈显著负相关.免疫印迹(Western blot)结果显示AML细胞株中KCTD5蛋白表达量显著高于正常对照细胞.该文通过shRNA干扰技术,敲低U937细胞中KCTD5的表达,研究了KCTD5对人急性单核细胞白血病U937细胞增殖的影响.采用CCK8实验、锥虫蓝拒染实验、瑞氏染色、PI单染及Western blot技术,检测干扰KCTD5对U937细胞生长、活力、细胞形态、细胞周期分布及周期相关蛋白表达的影响.结果 显示,干扰KCTD5的表达可以显著抑制U937细胞的增殖能力,但对细胞活力没有明显影响;形态学检查显示,干扰KCTD5后,U937细胞体积明显增大,染色质凝集及多核细胞比例增加;流式结果显示,干扰KCTD5的表达使G2/M期细胞比例明显增加;Western blot结果显示,干扰KCTD5后,Cdk1Tyr15磷酸化水平下降,而Cyclin B1表达量升高.以上结果表明,干扰KCTD5可以抑制人急性单核细胞白血病U937细胞增殖,其机制可能是由于干扰KCTD5通过使CdklTyr15发生去磷酸化,从而诱导U937细胞发生了G2/M期阻滞.

乳腺癌是一种常见的高度恶性肿瘤,在临床病人的治疗中,寻找有效的抑制肿瘤生长的治疗方法尤为重要.他莫昔芬目前被用于治疗雌激素受体阳性乳腺癌.二甲双胍是一种抗糖尿病药物,据报道可以降低人类癌症发病率,提高乳腺癌患者的生存率.该文主要研究二甲双胍联合他莫昔芬对乳腺癌细胞的协同作用及其机制.采用CCK-8法和平板克隆形成实验检测细胞活力和增殖;流式细胞术检测细胞凋亡;Transwell实验检测细胞迁移、侵袭能力;免疫印迹法检测MAPK信号通路和c-Myc蛋白.结果 显示,他莫昔芬与二甲双胍联合用药对乳腺癌细胞增殖、克隆形成、迁移侵袭及凋亡的作用均优于单独用药,表明二甲双胍可以增强他莫昔芬对肿瘤生长的抑制作用,并能下调c-Myc蛋白的表达.该研究结果显示,二甲双胍能明显提高乳腺癌细胞的抗肿瘤作用,这些影响是通过下调c-Myc蛋白介导的.该发现可能对乳腺癌的治疗有潜在的临床应用价值.

该文研究了低表达高迁移率族蛋白(high mobility group box 1,HMGB1)对结直肠癌细胞迁移侵袭的影响,构建了稳定低表达HMGB1的结直肠癌细胞系SW480和HCT-15.采用Transwell实验检测细胞迁移和侵袭能力,免疫印迹法(Western blot)检测蛋白表达水平.Transwell实验结果表明,shHMGB1组(HMGB1低表达组)的迁移侵袭能力明显强于shNC组(阴性对照组)(**P＜0.01,***P＜0.001).Western blot结果显示,与shNC组相比,shHMGB1组上皮标志物E-Cadherin蛋白的表达水平下降(*P＜0.05,**P＜0.01),间质标志物Snail蛋白的表达水平上升(*P＜0.05).此外shHMGB1组的c-Myc和GSK3B蛋白表达水平明显高于shNC组(*P＜0.05,**P＜0.01,***P＜0.001).上述结果表明,低表达HMGB1能显著促进结直肠癌细胞SW480和HCT-15的迁移和侵袭能力以及上皮-间质化(epithelial-mesenchymal transition,EMT),这为结直肠癌的治疗提供了新的潜在靶点和视角.

Mitochondrial disease was a clinically and genetically heterogeneous group of diseases, thus the diagnosis was very difficult to clinicians. Our objective was to analyze clinical and genetic characteristics of children with mitochondrial disease in China. We tested 141 candidate patients who have been suspected of mitochondrial disorders by using targeted next-generation sequencing (NGS), and summarized the clinical and genetic data of gene confirmed cases from Neurology Department, Beijing Children's Hospital, Capital Medical University from October 2012 to January 2015. In our study, 40 cases of gene confirmed mitochondrial disease including eight kinds of mitochondrial disease, among which Leigh syndrome was identified to be the most common type, followed by mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS). The age-of-onset varies among mitochondrial disease, but early onset was common. All of 40 cases were gene confirmed, among which 25 cases (62.5%) with mitochondrial DNA (mtDNA) mutation, and 15 cases (37.5%) with nuclear DNA (nDNA) mutation. M.3243A>G (n=7) accounts for a large proportion of mtDNA mutation. The nDNA mutations include SURF1 (n=7), PDHA1 (n=2), and NDUFV1, NDUFAF6, SUCLA2, SUCLG1, RRM2B, and C12orf65, respectively.

Objective To analyze the relationship between neutrophil/lymphocyte ratio in peripheral blood and inflammation degree in mice infected by Escherichia coli,and the possible mechanism of action of neutrophil and lymphocyte upon being infected by Escherichia coli.Methods 44 ICR mice were injected with 1 × 109 CFU/mL Escherichia coli to build the models,with 4 mice as the blank controls and 4 as the negative controls.The quantities of white blood cells,neutrophils,lymphocytes and neutrophil/lymphocyte ratio were detected by using SYSMEX2100 at eleven time points.Results The quantity of white blood cells in the model group was lower than that in control group (0.01＜P＜0.05) in 1 to 24 hours after infection,showed no obvious difference from that in control group on the second day,was significantly higher than that in control group from the third day to the seventh day,and then declined on the seventh day.The quantity of neutrophils was statistically higher than that in control group from the 12th hour to 7th day.The neutrophil/lymphocyte ratio in control group was 0.06+ 0.03,not obviously different from the 0.07 ±0.04 in the model group at 1 hour after infection (P＞0.05) but from those at 3 hours and 6 hours after infection (0.01＜ P＜0.05);and the ratios in the model group were significantly different from that in the control group from the 12th hour to the 7th day after infection,which reached the highest value at Day 7 (1.52 + 0.38).Conclusion Compared to quantities of white blood cells and neutrophil,neutrophil/lymphocyte ratio is a more sensitive indicator for Escherichia coli infection in mice,and when used in combination with the former,it can provide more sensitive reference data for the clinical diagnosis of Escherichia coli infection.

Objective To optimize the in utero electroporation protocol for transfering genes into the embryonic murine brain. Methods Critical steps of in utero electroporation were optimized, including choosing isoflurane anesthesia, improving the preparation of micropipettes, reducing the exposure time of embryos during the surgery as well as optimizing electroporation parameters. Forty-two mouse embryos from 6 mothers were subjected to in utero electroporation at gestational day 16. 5 using the optimized protocol. Results The survival rates of mothers and embryos were 100％. The tranfered gene was expressed in 85％ ± 3％ of electroporated embryonic brains. Conclusion Using the optimized in utero electroporation protocol, a satisfying survival rate of embryos and transfection efciency can be achieved.

Objective To establish and evaluate a rat model of inhalation lung injury induced by ship smog.Methods A rat model of inhalation lung injury was established by analyzing the composition of ship materials after combustion.Fortytwo healthy male Wistar rats were randomly divided into normal control group and 2,6,12,24,48 and 72h groups (6 each)after inhalation,these rats were killed at each time point,and the changes of arterial blood gas,coagulation function,the lung water content (％) were detected.Macroscopic and microscopic changes in lung tissues were observed to judge the degree of lung injury.Results The main components after combustion of 7 kinds of nonmetal materials on ship included CO,CO2,H2S,NOx and other harmful gases in this study,AIKE in one gas detector was used to monitor O2,CO,CO2 and H2S,and their concentrations remained relatively stable within 15 minutes,and the injury time was 15 minutes.The rats presented with shortness of breath and mouth breathing.Smoke inhalation caused a significant hypoxemia,the concentration of blood COHb reached a peak value 2h and the lung water content (％) did 6h after inhalation (P＜0.05).It is metabolic acidosis in the early stage after inhalation,but metabolic acidosis combined with respiratory acidosis in the later period.Histopathological observation showed diffuse hemorrhage,edema and inflammatory cell infiltration in the lung tissue as manifestations of lung injury,and the injury did not recover at 72h after inhalation,the change of blood coagulation function was not statistically significant.Conclusion A rat model of inhalation lung injury induced by ship smog has been successfully established,and has the advantages of easy replication,stability and reliability,thus can be used to research and treat inhalation lung injury induced by ship smog in naval war environment and other cases.

目的 探讨吡非尼酮(PFD)对舰船烟雾所致吸入性肺损伤大鼠的防护作用.方法 将健康成年雄性Wistar大鼠放入自制发烟装置,吸入舰船烟雾15 min建立吸入性肺损伤大鼠模型.吸入烟雾前30 min,正常对照组大鼠和烟雾吸入损伤组分别ip含5％二甲亚砜的生理盐水2 mL,PFD组ip PFD 300 mg·kg-1(溶于5％二甲亚砜的生理盐水)2 mL.分别于12,24和48 h每组各取6只大鼠,腹主动脉取血,检测动脉血气.双抗体夹心法检测白细胞介素6(IL-6)、IL-17和肿瘤坏死因子α(TNF-α)水平;取右肺前叶,计算肺含水率;留取右肺中叶,检测肺组织中丙二醛(MDA)含量;取右肺下叶经4％甲醛固定后做病理切片,苏木精-伊红染色,光镜观察.结果 与正常对照组比,舰船烟雾吸入损伤暴露后12,24和48 h,烟雾吸入损伤组各时相点O2分压(paO2)降低,CO2分压(paCO2)升高(P<0.05),肺含水率水平升高(P<0.05),血清中IL-6,IL-17和TNF-α水平升高(P<0.05),肺组织匀浆中MDA含量增加(P<0.05).与烟雾吸入损伤组相比,PFD组大鼠动脉血中paO2水平升高,paCO2及肺含水率降低(P<0.05),大鼠血清中IL-6,IL-17和TNF-α水平降低,组织中MDA含量降低(P<0.05).病理观察可见,PFD组较舰船烟雾吸入损伤组肺组织水肿减轻,肺泡腔渗出及炎症细胞浸润减少.结论 PFD可通过抑制IL-6等相关炎症因子的释放减轻舰船烟雾致大鼠吸入性肺损伤.

线粒体病具有高度临床异质性和基因异质性,对于临床医生来说诊断非常困难.本研究分析了中国线粒体病患儿的临床特征和基因特点,为临床医生的诊断提供了一定的参考依据.本文总结了2012年10月~2015年1月就诊北京儿童医院的141例已进行二代测序基因检测的疑似线粒体病患儿,分析基因确诊病例的临床特征以及遗传学特点.本研究共收集了40例基因确诊病例,包括8种不同类型的线粒体病,而Leigh综合征是最常见类型,其次是线粒体脑肌病伴乳酸酸中毒和卒中样发作（MELAS）综合征.不同的线粒体病发病年龄各异,但是普遍临床早发.40例基因确诊病例中, 25例（62.5%）为mtDNA突变, 15例（37.5%）为nDNA突变. m.3243A＞G突变7例,在mtDNA突变中最为常见, nDNA突变包括SURF1 7例, PDHA1 2例, NDUFV1, NDUFAF6, SUCLA2, SUCLG1, RRM2B,以及C12orf65各一例.