丙泊酚复合瑞芬太尼用于无肌松药气管插管的临床研究

目的:探讨丙泊酚复合瑞芬太尼用于无肌松药气管插管的最佳剂量,评估诱导期应用此种方法的有效性。方法:择期全麻的患者100例,ASAⅠ~Ⅱ级,随机分为三组。麻醉诱导为丙泊酚2 mg/kg,瑞芬太尼为2μg/kg(Ⅰ组)、3μg/kg(Ⅱ组)和4μg/kg(Ⅲ组),待两种药物达到药效高峰时行气管插管。观察诱导期和插管后1 min,3 min的血流动力学的变化情况,评估插管满意度。结果:各组的插管满意度如下:Ⅰ组54.2%(19/35)、Ⅱ组83.3%(25/30)、Ⅲ组97.1%(34/35),随着瑞芬太尼剂量的增加,气管插管满意度增高,气管插管操作时间缩短(P0.05)。在诱导过程中,三组患者的MAP和HR下降幅度均较显著(P0.05)。结论:丙泊酚复合瑞芬太尼用于无肌松药气管插管是可行的,与复合瑞芬太尼2μg/kg或3μg/kg相比,丙泊酚2 mg/kg复合瑞芬太尼4μg/kg能明显提供更好的气管插管条件,且插管操作时间明显缩短。     

大鼠气管插管方法学概述

由于大鼠呼吸频率较快、口腔狭小、声门较高,医学实验中气管内插管操作具有较多困难,多年来很多学者对大鼠气管内插管方法进行了大量研究。本文主要对大鼠气管内插管时动物和气管导管的选择、麻醉方式、插管的体位以及各种插管工具和方法等作一简要的综述。     

靶控输注依托咪脂在老年患者颌面外科手术中的应用效果

目的:研究靶控输注依托咪脂在老年患者颌面外科手术中的应用效果。方法:选择60例美国麻醉医师协会(ASA)分级I～II级行颌面外科手术的患者,并将其随机分为靶控输注丙泊酚组(P组,n=30)和靶控输注依托咪酯组(E组,n=30),记录麻醉诱导前、气管插管前、气管插管后5 min、手术开始时、停药时的有创收缩压、有创舒张压、心率,并记录手术时间、拔管时间、麻醉维持时依托咪酯和丙泊酚的靶控输注浓度。结果:P组气管插管前、气管插管后5 min、手术开始时、停药时血压均低于麻醉诱导前,且低于E组各对应时间点,差异均具有统计学意义(P0.05),P组气管插管前心率显著低于麻醉诱导前和E组(P0.05);E组气管插管前、气管插管后5 min和手术开始时血压均低于麻醉诱导前(P0.05),但各时间点心率无显著变化(P0.05)。P组靶控浓度为(1.95±0.54)μg/m L～(4.48±0.61)μg/mL,E组靶控浓度为(0.20±0.05)μg/mL～(0.51±0.05)μg/mL。两组药物拔管时间、术后恶心呕吐和躁动发生率差异无统计学意义(P0.05),但P组低血压发生率显著高于E组(P0.05)。结论:在颌面外科手术中,靶控输注依托咪脂,能有效减少麻醉诱导和维持中低血压发生,维持血流动力学稳定,对于老年患者而言,是一种比丙泊酚更为安全可靠的静脉麻醉药。     

大气细颗粒物致炎动物模型系统评价

目的 探索大气污染对动物的致病机制,对BALB/c小鼠采用无创性气管滴注PM2.5颗粒悬浮液的方法,构建大气污染致炎动物模型。方法 将150只SPF级BALB/c小鼠随机分成空白对照组、生理盐水组、PM2.5低度组(2.5 mg/kg)、PM2.5中度组(5 mg/kg)和PM2.5高度组(10 mg/kg)共5组,各剂量组气管滴注第3天,第7天、第21天、第35天、第49天,气管滴注操作完成后24 h采取组织样本,采用ELISA、肺组织病理HE染色的方法,来验证无创性气管滴注方法的可行性和致炎模型构建成功与否。结果 本建模方法,成功率高达96%。采用气管滴注法,建模小鼠肺组织炎症评分与气道滴注时间的延长和剂量呈正相关。PM2.5暴露后,肺内有大量淋巴细胞聚集及吞噬颗粒的巨噬细胞浸润,肺泡间隔增宽。各暴露组分别与生理盐水对照组、空白组比较,肺泡灌洗液中炎症因子IL-6、肺组织匀浆中TNF-α水平增高,高剂量组差异最显著。结论 本实验用气管滴注法建立小鼠致炎模型成功,并证明此方法简单、可靠,可广泛用于小鼠呼吸系统重复滴注,有利于进一步研究大气污染及其他致炎机制。     

三种关于建立家兔人工气道方法的评价

目的比较关于家兔三种人工气道（artificial airway,AA）建立方法,为实验中快速建立家兔人工气道选择较好的方法提供参考。方法选用新西兰大耳白兔共30只,随机均分为3组。分别采用气管切开法、经口明视气管插管法、经口盲探气管插管法建立人工气道,比较各组建立人工气道的时间,首次成功率及并发症发生情况。结果经口盲探气管插管法建立人工气道的时间明显短于经口明视气管插管法和气管切开法,首次成功率由高到低依次是气管切开法、经口盲探气管插管法、经口明视气管插管法。经口盲探气管插管组并发症发生率最低,为（10±5）%,气管切开组和经口明视气管插管组并发症发生率分别为（20±10）%,（30±10）%。结论经口盲探气管插管是现阶段建立家兔人工气道时值得优先考虑的方法,简便易行、可操作性强,成功率高。     

大鼠肺纤维化形成中肺内肥大细胞结缔组织生长因子的表达

目的：观察博莱霉素（BLM）诱导肺纤维化形成中肺肥大细胞（MCs）是否表达结缔组织生长因子（CTGF）。方法：32只雄性SD大鼠,随机分为博莱霉素（BLM）组和对照（Control）组（n=16）。BLM组为气管内一次性滴注BLM（5mg／ks）;Control组为气管内滴注与BLM等容量的生理盐水（NS）。各组分别在气管滴注后第14天和第28天处死大鼠,取肺组织样本。用氯胺-T法检测肺组织羟脯氨酸含量以判断肺纤维化程度;用甲苯胺蓝染色显示肺组织切片中的MCs;免疫组化染色显示肺CTGF的表达和分布。结果：①与对照大鼠比,气管内滴注BLM后第28天大鼠的肺羟脯氨酸含量明显增高（P〈0．01）。②与对照大鼠比,气管内滴注BLM后第14天和第28天大鼠肺内MCs数明显增多（均P〈0．01）,肺内CTGF表达上调（均P〈0．01）。③对照大鼠肺内未见CTGF免疫阳性的MCs;而气管内滴注BLM后第14天和第28天大鼠肺内病灶区中有CTGF免疫阳性的MCs。结论：肺纤维化形成中肺MCs表达CTGF,这可能是MCs促进肺纤维化的作用机制之一。     

成年大鼠气管插管方法的改良

目的寻找一种较好的成年大鼠气管插管方法;方法对比明视经口气管内插管法与逆行导管引导插管法的成功率及并发症;结果明视经口气管内插管法的成功率为100%,逆行导管引导插管法的成功率为87%。插管后并发症明显低于逆行导管引导插管法。结论明视经口气管内插管法优于逆行导管引导插管法,值得推广应用。     

黄芩苷防止肺纤维化大鼠肺内结缔组织生长因子的上调

为探讨黄芩苷(baicalin,Bai)防止肺纤维化的机制,本研究观察了Bai对肺纤维化大鼠肺内结缔组织生长因子(connective tissue growth factor,CTGF)上调的影响。将雄性Sprague-Dawley(SD)大鼠随机分为4组:生理盐水(normal saline,NS)+NS组(气管内滴注NS,随后每天腹腔注射NS一次)、NS+Bai组(气管内滴注NS,随后每天腹腔注射Bai一次)、博莱霉素(bleomycin,BLM)+NS组(气管内滴注BLM,随后每天腹腔注射NS一次)和BLM+Bai组(气管内滴注BLM,随后每天腹腔注射Bai一次)。Bai的剂量分别为每天6、12.5或50mg/kg。各组于气管内一次性滴注BLM或NS后第28天处死动物,取肺组织样本。采用氯胺-T法检测肺组织羟脯氨酸含量(反映肺纤维化程度的指标),用RT-PCR和免疫组织化学方法检测肺CTGF的表达。结果显示,BLM+NS组大鼠肺羟脯氨酸含量、CTGF蛋白及mRNA水平均明显高于NS+NS组大鼠(均P0.01),提示BLM诱导了大鼠的肺纤维化,且纤维化肺内出现CTGF表达的上调。BLM+Bai组大鼠连续28d每天腹腔注射6、12.5或50mg/kgBai后,BLM所致的肺纤维化明显减轻,同时肺CTGF表达的上调也得到明显抑制。以上结果提示,Bai可防止肺纤维化大鼠肺内CTGF表达的上调,这可能是其防止肺纤维化形成的作用机制之一。     

PM2.5暴露对大鼠心脏功能及NLRP3炎性小体表达的影响

目的 探究气道吸入不同浓度PM2.5混悬液是否能激活大鼠心肌组织NLRP3炎性小体引起心脏功能的损伤,为寻找相关药物干预提供参考。方法 SD大鼠随机分为对照组,低(7.5 mg/kg)、中(15 mg/kg)、高(30 mg/kg)剂量染毒组,每组7只。染毒组通过非暴露式气管滴注相应剂量混悬液(1 mL/kg),6 d 1次,持续2个月,对照组大鼠滴注等量生理盐水,滴注期间每日记录大鼠生理状况。末次滴注完成后,禁食12 h, 1%戊巴比妥钠(50 mg/kg)麻醉,以心肌组织病理切片、心肌细胞凋亡状况、心肌谱酶变化反应心脏功能状况;以心肌组织Bcl-2,Bax蛋白;NLRP3、Caspase-1、IL-1β蛋白及mRNA表达,血清中IL-1β、IL-6、TNF-α、IL-18表达水平反应心肌组织NLRP3炎性小体的活化状况。结果 滴注不同剂量PM2.5混悬液后,各染毒组大鼠不同程度毛色变黄、无光泽,活动减少,饮食、体重无明显影响,心肌组织出现不同程度的横纹断裂、细胞减少、排列无规则、细胞核固缩、水肿的现象。与对照组相比,染毒组大鼠心肌组织中CK、LDH、AST水平,Bcl-2、IL-1...     

肺纤维化大鼠模型造模方法的优化

目的 比较气管内滴注、气管内雾化喷入博来霉素(5 mg/kg)两种给药方式,以及比较腹腔注射3%戊巴比妥钠麻醉、异氟烷呼吸麻醉(与氧气混合吸入时,异氟烷浓度为0.5%)两种麻醉方式对肺纤维化大鼠模型的影响,探讨更优的造模方法。方法 选择雄性SPF级SD大鼠50只,随机分为空白对照组、腹腔麻醉气管内滴注组、腹腔麻醉气管内雾化喷入组、呼吸麻醉气管内滴注组和呼吸麻醉气管内雾化喷入组,每组各10只。观察给药后1、3、7、14、21 d,各组大鼠的生存状况及体重变化;给药3周后处死大鼠,取肺称重,计算肺系数;HE染色观察肺组织炎症变化;Masson染色观察肺组织中胶原增殖;Western Blot检测肺组织中转化生长因子-β1(TGF-β1)蛋白表达量;碱水解法检测肺组织中羟脯氨酸(HYP)的含量。结果 与空白对照组比,4种模型组的大鼠精神状态不佳、体重下降、肺指数上升;肺组织损伤明显,炎症水平增加,胶原增殖显著;肺组织中TGF-β1蛋白表达水平升高(P<0.001);发现仅气管内雾化喷入博来霉素组大鼠肺组织中羟脯氨酸的含量增加(P<0.05)。发现气管内滴注的建模方式其模型大鼠肺部...     

Baicalin prevents the up-regulation of connective tissue growth factor in fibrotic lungs of rats

To clarify the mechanism underlying the preventive effect of baicalin (Bai) on fibrosis in lung, we investigated the influence of Bai on the up-regulation of connective tissue growth factor (CTGF) in fibrotic lungs. Male Sprague-Dawley (SD) rats were divided into four groups randomly: normal saline (NS)+NS group (a single intratracheal instillation of NS plus i.p. injection of NS), NS+Bai group (intratracheal instillation of NS plus i.p. injection of Bai), bleomycin (BLM)+NS group (intratracheal instillation of BLM plus i.p. injection of NS) and BLM+Bai group (intratracheal instillation of BLM plus i.p. injection of Bai). All the i.p. injections were performed once daily. On day 28 after intratracheal instillation of BLM or NS, the rats were sacrificed for lung tissue sampling. As the index of the severity of pulmonary fibrosis, the content of hydroxyproline in lungs was analyzed by chloramine T method. The expression levels of CTGF mRNA and protein in the lungs were detected by RT-PCR and immunohistochemistry, respectively. The results showed that, compared to the rats in NS+NS group, the rats in BLM+NS group showed increased hydroxyproline content and higher levels of CTGF mRNA and protein expressions (P<0.01), suggesting that BLM had induced fibrosis in lung and up-regulated CTGF expression in the fibrotic lungs. Administration of different dosages of Bai (6, 12.5 and 50 mg/kg per d, for 28 days) into the BLM-treated rats reduced the increased content of hydroxyproline, and ameliorated the up-regulation of CTGF mRNA and protein levels, respectively. These results suggest that Bai could prevent the up-regulation of CTGF expression in fibrotic lungs of rats receiving BLM instillation, which might be one of the mechanisms underlying the preventive effect of Bai on pulmonary fibrosis.     

Development of a new aerosol delivery system for systemic pulmonary delivery in anaesthetized and orotracheal intubated rats.

Over the last decade, the systemic absorption of a broad range of therapeutics after pulmonary application has been demonstrated in animals as well as in humans. The most common method used in the laboratory is the intratracheal instillation of drugs in solution. This method is, however, unsatisfactory, because of discrepancies in particle distribution, clearance, kind of injury and bioavailability between instillation and inhalative application. On the other hand, a precise determination of the amount of drug applied by aerosol, and of the aerosol volume retained within the lungs is rather difficult, and is not possible for use with small animals such as mice or rats. We describe a system which allows the delivery of aerosols directly into the animal's lungs, and calculation of the amount of drug retained in the lungs. Our system was tested in vitro and in vivo and was shown to allow precise and efficient pharmacokinetic and toxicological studies to be carried out.     

A method for intratracheal instillation in the rat.

A method was developed to provide safe, rapid, and accurate intratracheal instillation of radionuclides into the rat lung. Rats were anesthetized with halothane and suspended vertically on an animal support stand with rubber bands attached to the incisor teeth, holding the mouth open. A speculum was inserted into the trachea and the radionuclide was injected from a syringe through tubing inserted through the tracheal speculum into the trachea. The amount of radionuclide retained in the lung varied by less than a factor of 2 within groups of rats observed 4 hr-20 da after employing this method of instillation. When a volume of 2 ml was instilled, the radionuclide distribution between the left and right lung was similar to that observed following incorporation by inhalation. Instillations of less than 2 ml showed unequal distribution of radionuclide between the right and left lung.     

Therapeutic Effect of Intravenous Infusion of Perfluorocarbon Emulsion on LPS-Induced Acute Lung Injury in Rats

Acute lung injury (ALI) and its more severe form, acute respiratory distress syndrome (ARDS) are the leading causes of death in critical care. Despite extensive efforts in research and clinical medicine, mortality remains high in these diseases. Perfluorocarbon (PFC), a chemical compound known as liquid ventilation medium, is capable of dissolving large amounts of physiologically important gases (mainly oxygen and carbon dioxide). In this study we aimed to investigate the effect of intravenous infusion of PFC emulsion on lipopolysaccharide (LPS) induced ALI in rats and elucidate its mechanism of action. Forty two Wistar rats were randomly divided into three groups: 6 rats were treated with saline solution by intratracheal instillation (control group), 18 rats were treated with LPS by intratracheal instillation (LPS group) and the other 18 rats received PFC through femoral vein prior to LPS instillation (LPS+PFC group). The rats in the control group were sacrificed 6 hours later after saline instillation. At 2, 4 and 6 hours of exposure to LPS, 6 rats in the LPS group and 6 rats in LPS+PFC group were sacrificed at each time point. By analyzing pulmonary pathology, partial pressure of oxygen in the blood (PaO2) and lung wet-dry weight ratio (W/D) of each rat, we found that intravenous infusion of PFC significantly alleviated acute lung injury induced by LPS. Moreover, we showed that the expression of pulmonary myeloperoxidase (MPO), intercellular adhesion molecule-1 (ICAM-1) of endothelial cells and CD11b of polymorphonuclear neutrophils (PMN) induced by LPS were significantly decreased by PFC treatment in vivo. Our results indicate that intravenous infusion of PFC inhibits the infiltration of PMNs into lung tissue, which has been shown as the core pathogenesis of ALI/ARDS. Thus, our study provides a theoretical foundation for using intravenous infusion of PFC to prevent and treat ALI/ARDS in clinical practice.     

Metabolism of testosterone to 17 beta-hydroxy-5 alpha-androstane-3-one and 5 alpha-androstane-3 alpha, 17 beta-diol in alveolar macrophages from rat lung--II. Effects of intratracheal instillation of 3.4 mumol K2Cr2O7

Activity of the steroid 5 alpha-reductase in pulmonary alveolar macrophages from adult male rats has been investigated in vitro. Intratracheal instillation of 3.4 mumol K2Cr2O7 lowered the enzyme activity within 6 h, and the reduction was significant on the subsequent 2, 4 and 7 days. The activity of this enzyme was significantly decreased only 6 and 24 h after instillation when measured in the 800 g supernatant fraction of whole lung. Instillation of 3.4 mumol K2Cr2O7 increased serum levels of corticosterone. Serum levels of triiodothyronine and thyroxine decreased except for a transient increase 3 h after the K2Cr2O7 instillation. Subcutaneous administration of 200 micrograms dexamethasone/100 g b.wt, 200 micrograms/100 g b.wt of testosterone, 17 beta-hydroxy-5 alpha-androstane-3-one (5 alpha-DHT), dehydroepiandrosterone or corticosterone had no effect on the 5 alpha-reductase activity of the pulmonary alveolar macrophages within 12 h. The combined treatment with dexamethasone s.c. and intratracheal instillation of 3.4 mumol K2Cr2O7 reduced the steroid 5 alpha-reductase activity in the pulmonary alveolar macrophages to about 25% of controls. Measurement of the steroid 5 alpha-reductase activity in pulmonary alveolar macrophages as an index of lung damage when exposed to toxic material is discussed.     

气管内反复滴入脂多糖法建立大鼠慢性阻塞性肺疾病模型

目的评价脂多糖(LPS)诱发大鼠慢性阻塞性肺疾病(COPD)模型的可行性。方法气管内滴入脂多糖或生理盐水,每周1次,共8周。测定大鼠的气道阻力(RL)和肺动态顺应性(Cdyn),计数大鼠支气管肺泡灌洗液(BALF)中白细胞总数及分类,肺组织病理切片行HE和AB-PAS染色,并测定肺组织粘蛋白(MUC5AC)的含量。结果模型组大鼠RL明显升高(87.5%),Cdyn显著下降(16.4%);BALF中白细胞总数及分类中的中性粒细胞、淋巴细胞和单核巨噬细胞数均明显高于对照组;光镜下可见病变呈慢性支气管炎及肺气肿样改变。结论反复气管内滴入LPS可用于制备大鼠COPD模型,其肺功能、BALF细胞学及病理学改变符合人类COPD表现,可以用于实验研究。     

A simple non-surgical method of intrabronchial instillation for the establishment of respiratory infections in the rat.

A technique involving intubation of weanling rats for intrabronchial instillation of infectious material has been developed to produce a respiratory infection model suitable for experimental infection studies. The technique is performed simply with the use of a metal cannula and an intravenous plastic cannula, and dispenses with the need for illumination, laryngoscopy and extensive manipulation of the animal.     

吸入糖皮质激素对大鼠肺纤维化的干预作用及bFGF表达的影响

目的:研究吸入糖皮质激素对大鼠肺纤维化模型的干预作用及可能的机制。方法:雌性Wistar大鼠40只,体重180～250g,按照随机数字表法将大鼠随机分为4组(n=10):①对照组(C组);②模型组(M组);③布地奈德组(B组);④地塞米松组(D组)。M组、B组、D组给大鼠气管内吸入博莱霉素(5mg/kgbw,8mg)复制肺纤维化模型,C组气管内吸入同等剂量的生理盐水作对照,B组于次日给予雾化吸入等效剂量布地奈德,D组于次日腹腔内注射地塞米松。上述各组均于注药后第1、4周各宰杀5只。通过苏木素-伊红染色观察肺泡炎、Masson胶原染色观察肺纤维化、用免疫组化及酶联免疫吸附测定(ELISA)法检测bFGF蛋白在大鼠肺组织,血清及肺泡灌洗液(BALF)的表达。结果:1、4周时M组表现为肺泡炎及肺间质炎症,B组、D组肺泡炎及肺纤维化程度较M组减轻。1、4周时血清、肺组织、BALF中M组的bFGF表达高于C组(P0.01),B、D组低于M组(P0.01)。结论:吸入糖皮质激素可减轻博莱霉素诱导的肺纤维化,其抗纤维化作用的机制与抑制bFGF表达有关。     

Intubation-mediated Intratracheal (IMIT) Instillation: A Noninvasive,Lung-specific Delivery System

Respiratory disease studies typically involve the use of murine models as surrogate systems. However, there are significant physiologic differences between the murine and human respiratory systems, especially in their upper respiratory tracts (URT). In some models, these differences in the murine nasal cavity can have a significant impact on disease progression and presentation in the lower respiratory tract (LRT) when using intranasal instillation techniques, potentially limiting the usefulness of the mouse model to study these diseases. For these reasons, it would be advantageous to develop a technique to instill bacteria directly into the mouse lungs in order to study LRT disease in the absence of involvement of the URT. We have termed this lung specific delivery technique intubation-mediated intratracheal (IMIT) instillation. This noninvasive technique minimizes the potential for instillation into the bloodstream, which can occur during more invasive traditional surgical intratracheal infection approaches, and limits the possibility of incidental digestive tract delivery. IMIT is a two-step process in which mice are first intubated, with an intermediate step to ensure correct catheter placement into the trachea, followed by insertion of a blunt needle into the catheter to mediate direct delivery of bacteria into the lung. This approach facilitates a >98% efficacy of delivery into the lungs with excellent distribution of reagent throughout the lung. Thus, IMIT represents a novel approach to study LRT disease and therapeutic delivery directly into the lung, improving upon the ability to use mice as surrogates to study human respiratory disease. Furthermore, the accuracy and reproducibility of this delivery system also makes it amenable to Good Laboratory Practice Standards (GLPS), as well as delivery of a wide range of reagents which require high efficiency delivery to the lung.