目的 基于mRNA测序(RNA-seq)技术,采用生物学信息分析软件分析清瘟败毒饮对脓毒症大鼠心肌组织中白细胞介素-17(IL-17)信号转导通路相关基因表达的变化,阐明清瘟败毒饮对脓毒症心肌的保护机制.方法 清洁级健康雄性Wistar大鼠45只,随机分为脓毒症模型组、清瘟败毒饮组和假手术组,每组15只.采用盲肠结扎穿孔术(CLP)制作脓毒症大鼠模型,假手术组仅开腹、关腹与复苏,不进行CLP.清瘟败毒饮组大鼠在制模前2?d给予清瘟败毒饮(组成:生地黄7.5?g,生石膏、水牛角各15?g,黄连2.5?g,栀子、桔梗、黄芩、知母、赤芍、玄参、连翘、甘草、丹皮、竹叶各5?g)胃饲,每日2次,术后继续中药灌胃24?h.3组术毕均予5?mL/kg平衡液肌肉注射.术后24?h取大鼠肌组织提取总RNA,采用RNA-seq技术对mRNA表达量进行检测,并应用京都基因和基因组数据库通路富集方法(KEGG?Pathway)分析清瘟败毒饮对脓毒症大鼠心肌组织IL-17信号通路相关基因表达的影响.结果 RNA-seq检测结果显示,与假手术组相比,脓毒症模型组大鼠心肌组织表达上调的基因有900个,下调的基因有1507个;清瘟败毒饮组大鼠心肌组织表达上调的基因有1288个,下调的基因有1870个.KEGG?Pathway分析结果显示,与假手术组相比,脓毒症模型组大鼠心肌组织IL-17信号通路相关基因表达上调的有10个,下调的有5个;清瘟败毒饮组大鼠心肌组织IL-17信号通路基因表达上调的有11个,下调的有6个.其中脓毒症模型组及清瘟败毒饮组同时上调的基因有7个〔CXC趋化因子配体(CXCL1、CXCL2)、白细胞介素-6(IL-6)、环氧合酶-2(COX-2)、粒细胞集落刺激因子(G-CSF)、基质金属蛋白酶9?(MMP-9)、CCAAT/增强子结合蛋白β(C/EBPβ)〕;同时下调的基因有5个〔转化生长因子激酶1(TAK1)、核转录因子-κB(NF-κB)、泛素特异性蛋白酶25(USP25)、细胞外信号调节激酶(ERK)、天冬氨酸特异性半胱氨酸蛋白酶(CASP)〕;脓毒症模型组上调而清瘟败毒饮组正常表达的基因有2个〔CXCL5、白细胞介素-1β(IL-1β)〕;?脓毒症模型组下调而清瘟败毒饮组正常表达的基因有1个〔热休克蛋白90(HSP90)〕;脓毒症模型组正常表达而清瘟败毒饮组上调的基因有3个〔激活蛋白-1(AP-1)、IκB激酶-β(IKK-β)、CC趋化因子7(CCL7)〕;脓毒症模型组正常表达而清瘟败毒饮组下调的基因有1个(ERK);脓毒症模型组上调而清瘟败毒饮组下调的基因有1个〔IκB激酶-α(IKK-α)〕;脓毒症模型组下调而清瘟败毒饮组上调的基因有1个〔NF-κB抑制因子-α(IκB-α)〕.结论 清瘟败毒饮可通过调节IL-17信号通路相关基因的表达,对脓毒症大鼠心脏起保护作用.

目的 观察瑞芬太尼对脓毒症大鼠能量代谢指标乳酸、丙酮酸、肾上腺素表达的影响,为瑞芬太尼治疗脂多糖( LPS)诱导的脓毒症提供理论依据. 方法 采用雄性清洁级Wistar大鼠54只,随机分为3组,即对照组21只、LPS组18只,瑞芬太尼处理组15只. LPS组于20 min内尾静脉注射LPS 15 mg/kg制作脓毒症模型,对照组应用相同量的生理盐水代替;瑞芬太尼处理组,先注射等量 LPS后再给予瑞芬太尼(0.04 mg/kg)尾静脉泵入40 min.测定各组大鼠造模后6 h的心、肾组织能量代谢指标表达情况,采用生化法检测乳酸、丙酮酸水平,ELISA检测肾上腺素水平. 结果 与对照组比较,LPS组大鼠心、肾组织的乳酸、丙酮酸、肾上腺素水平明显增加( P＜0.05) ,而瑞芬太尼显著改善上述指标的变化. LPS组大鼠心、肾组织的乳酸、丙酮酸水平与肾上腺素水平呈正相关(P＜0.05). 结论 瑞芬太尼可通过调节机体能量代谢,减轻能量代谢紊乱所致的组织缺氧,保护LPS所致的脓毒症组织损伤.

目的观察瑞芬太尼对脓毒症所致急性肺损伤（ALI）大鼠肺湿/干质量比、肺组织病理学改变及炎症因子及丙二醛（MDA）和超氧化物歧化酶（SOD）的影响,为瑞芬太尼治疗脂多糖（LPS）诱导的ALI提供理论依据。方法采用雄性清洁级Wistar大鼠32只,体质量270～320g,随机分为四组:对照组、ALI组、瑞芬太尼处理组和瑞芬太尼对照组。ALI组于20min内尾静脉注射LPS 15mg/kg制作脓毒症ALI模型,对照组用相同量的生理盐水代替;瑞芬太尼对照组和瑞芬太尼处理组,先注射等量生理盐水/LPS,后给予瑞芬太尼（0.04mg/kg）尾静脉泵入40min。测定各组大鼠造模后6h后的肺湿/干比;苏木精-伊红（HE）染色方法对肺组织进行病理观察,ELISA检测肿瘤坏死因子（TNF-α）,白介素-6（IL-6）的水平,并测定SOD活性及MDA的含量。结果与对照组比较,ALI组大鼠肺干湿比、肺组织TNF-α、IL-6水平、MDA含量增加而SOD活性显著降低（P ＜0.05）;而瑞芬太尼显著改善上述指标的变化。肺组织病理学观察发现ALI大鼠肺组织结构损伤严重,可见大量炎性细胞聚集。而瑞芬太尼处理组大鼠的上述病理学改变则明显减轻。结论瑞芬太尼可以通过抑制肺组织炎症因子的释放,减轻氧化应激途径保护LPS所致的急性肺损伤。

目的 探讨重症医学科(ICU)下呼吸道感染临床分离出的肺炎克雷伯杆菌对常用抗菌药物的耐药性,并探讨其对碳青霉烯类抗菌药物耐药的危险因素.方法 采用回顾性分析方法,分析2013-04～2015-04 ICU临床痰液标本中分离的肺炎克雷伯杆菌耐药性及碳青霉烯类耐药肺炎克雷伯杆菌(CRKP)感染的危险因素.结果 ①肺炎克雷伯杆菌对阿米卡星的敏感率最高,为86.81％,对亚胺培南和美罗培南的敏感率分别为62.5％和62.86％.153株肺炎克雷伯杆菌中,共检出CRKP 56株,检出率36.7％.CRKP对阿米卡星的敏感率仍较高,达68.52％,对复方新诺明的敏感率达25％,但对其他抗菌药物的敏感率均小于20％.耐药率严重.②单因素分析发现,肺炎克雷伯杆菌感染前住院天数≥5 d、住ICU≥7 d、机械通气≥7 d、同时存在真菌感染、2周内使用碳青酶烯类抗菌药物、留置尿管、WBC计数＞10.0×109/L或＜4.0×109/L及高血压病是ICU下呼吸道感染中肺炎克雷伯杆菌对碳青霉烯类抗菌药物产生耐药的危险因素(P＜0.05).Logistic回归分析发现,住院天数≥5 d、2周内使用碳青酶烯类抗菌药物是ICU下呼吸道感染中肺炎克雷伯杆菌耐碳青霉烯类抗菌药物的独立危险因素.结论 ICU内肺炎克雷伯杆菌耐药率严重,减少碳青霉烯类抗菌药物的使用,尽量缩短住院时间有利于预防ICU内CRKP所致下呼吸道感染的发生.

>鲍曼不动杆菌（acinetobacter baumannii,AB）是一种经常引起ICU感染暴发流行的革兰氏染色阴性杆菌,它涉及的感染可包含脓毒症、肺部感染、尿路感染、伤口感染、脑膜炎等重症感染 [1] 。近年来,鲍曼不动杆菌的感染率呈逐年上升的趋势,已成为院内感染常见的致病菌之一,而且,最令人担忧的是鲍曼不动杆菌能够聚集多种的耐药机制,产生对现有的所有抗菌药物耐药的菌株,是目前院内感染治疗上的一道难题 [2-4] 。因此,临床医

Objective To investigate gene expressions of GPR40 in βTC-3 cells induced by free fatty acids(FFA) and the intervention effect of pioglitazone(Piog) on those expressions.Methods βTC-3 cells studied in the experiment were subject to different treatments with a blank and three concentrations of FFA (0.25,0.5 and 1.0 mmol/L).Semi-quantitative RT-PCR analysis was used to determine the RNA expression level of GPR40 in βTC-3 cells after 12 hours and 24 hours respectively.Afterwards,βTC-3 cells were preincubated with different concentrations of Piog(0,0.1,1,10 μmol/L)for 6 hours,and 1 mmol/L FFA was then added and the cells were incubated for another 24 hours.The RNA expression level of GPR40 was determined by semi-quantitative RT-PCR at last.Results (1) After the cells were incubated with FFA of different concentrations for 12 hours,there was no statistically significant difference among all the groups.(2) After the 24-hour incubation,no difference was shown between the group with 0.25 mmol/L FFA and the control group.But compared to the control group and the group with 0.25 mmol/L FFA,the expression of GPR40 in βTC-3 cells in both groups with 0.5 mmol/L FFA and that with 1 mmol/L FFA decreased after the 24-hour incubation (P0.05).There was,however,no statistical difference between the group with 0.5 mmol/L FFA and that with 1 mmol/L FFA.(3)Though no statistically significant difference was found between the group with 0.1 μmol/L Piog+1.0 mmol/L FFA and that with 1.0 mmol/L FFA,the RNA expression level of GPR40 in both the group with 1 μmol/L Piog+1.0 mmol/L FFA and that with 10 μmol/L Piog+1.0 mmol/L FFA increased compared to the group with 1 mmol/L FFA (P0.01).Conclusion The expression of GPR40 in βTC-3 cells may be inhibited by long-term administration of FFA of high concentration,while Piog can help to protect the expression of GPR40 against FFA-induced impairment in βTC-3 cell.

Objective To investigate the changes in gene expression spectrum of heart tissue in septic rats by DNA microarrays. Methods Thirty male Wistar rats were randomly divided into sepsis group and control group with 15 rats in each group. Cecal ligation and puncture (CLP) was used to reproduce rat sepsis model, and the success of reproduction was confirmed by examining the heart tissue with transmission electron microscope. Gene expression spectrum was studied with oligonucleotide gene expression profile microarray that contained 22 523 rat cDNA clones to detect the changes in gene expression pattern of rat heart tissue 24 hours after CLP. Genes with fluorescent signal of Cy3/Cy5 of ratio average (RA)>2.0 or RA<0.5 were identified as differential genes, then those that high correlated to sepsis were screened by means of related computer software, and their relationship was analyzed. Results Electron microscopic examination of heart tissue demonstrated that the sepsis model was successfully reproduced. Compared to the controls, gene expression of 418 genes of heart tissue of septic rat were changed 24 hours after CLP, accounting for 1.86%, and among them 200 genes showed up-regulation, 218 genes with down-regulation. Among known functional genes, 84 genes up-regulated and 74 genes down-regulated. They were related with a range of genetic functions, such as acute stress reaction, signal transduction, immune response, energy metabolism related genes etc. Conclusion There are a series of changes in gene expression in heart tissue apparently induced by sepsis rat. DNA microarray technology provides a new tool for rapidly analyzing them.