烟曲霉暴露对支气管哮喘大鼠气道黏蛋白MUC2表达的影响

目的 研究烟曲霉暴露对支气管哮喘(简称哮喘)大鼠气道黏蛋白MUC2表达的影响.方法 70只雄性Wistar大鼠随机数字表法分为7组(每组10只):慢性哮喘(A)组、慢性哮喘+烟曲霉孢子吸入1周(B)组、3周(C)组和5周(D)组、慢性哮喘+生理盐水吸入(E)组、卵清白蛋白(OVA)致敏生理盐水激发(F)组和OVA致敏生理盐水激发+烟曲霉孢子吸入(G)组.测定各组大鼠的基础气道阻力和气道阻力变化率,ELISA法测定支气管肺泡灌洗液(BALF)中IL-13水平,RT-PCR法检测肺组织MUC2 mRNA水平,免疫组织化学染色测定气道上皮细胞MUC2表达强度.结果 D组大鼠基础气道阻力为(1.06±0.28) cm H2O·ml-1·s-1、气道阻力变化率为(85.69±5.68)％、BALF中IL-13为(197±34) mg/L、肺组织MUC2 mRNA/β-actin mRNA为(3.0±0.6)、气道上皮细胞MUC2表达强度(A值)为(871±70)均高于A组[(0.52±0.13) cm H2O· ml-1·s-1、(31.62±3.62)％、(54±9)mg/L、(1.8±0.4)、(202±36)]、B组[(0.54±0.14) cmH2O·ml-1·s-1、(35.90±2.62)％、(96±16) mg/L、(1.9±0.3)、(258±48)]、C组[(0.89±0.22) cm H2O· ml-1·s-1、(60.91±4.26)％、(136±22) mg/L、(2.3±0.5)、(546±54)]和E组[(60.91±4.26) cm H2O· ml-1· s-1、(31.64±3.47)％、(37±8)mg/L、(1.7±0.4)、(188±39)](P＜0.01或P＜0.05);A组高于F组[(0.33±0.08) cm H2O· ml-1·s-1、(16.85±3.62)％、(23±6)mg/L、(0.5±0.2)、(87±18)]和G组[(0.34±0.12) cm H2O· ml-l· s-1、(16.52±3.34)％、(25±6)mg/L、(0.5±0.1)、(88±15)](P＜0.01或P＜0.05);大鼠肺组织MUC2 mRNA/β-actin mRNA和气道上皮细胞MUC2表达强度(A值)均与BALF中IL-13水平正相关(r=0.648,P＜0.05;r=0.676,P＜0.05),与气道阻力变化率正相关(r=0.644,P＜0.05;r=0.584,P＜0.05).结论 慢性烟曲霉暴露可上调哮喘大鼠气道黏蛋白MUC2基因表达,气道黏液分泌增多,黏稠度和黏滞度增加,导致气道反应性增高.     

环孢素A对链脲佐菌素诱导的糖尿病大鼠肾脏基质金属蛋白酶-2和基质金属蛋白酶-9表达的影响

目的 观察免疫抑制剂环孢素A(CsA)对糖尿病大鼠肾脏基质金属蛋白酶(MMP)表达的影响.方法 健康雄性SD大鼠74只,平均体重250 g,采用数字表法随机分为正常对照组(n=10)、单纯糖尿病组(n=8)、胰岛素治疗组(n=9)、造模前低剂量CsA治疗组(n=9;造模前1周皮下注射1μg·g-1·d-1 CsA)、造模前中剂量CsA治疗组(n=8;造模前1周皮下注射4μg·g-1·d-1CsA)、造模前高剂量CsA治疗组(n=8;造模前1周皮下注射8μg·g-1·d-1CsA)、造模后低剂量CsA治疗组(n=8;造模后1周皮下注射1μg·g-1·d-1CsA)、造模后中剂量CsA治疗组(n=7;造模后1周皮下注射4μg·g-1·d-1 CsA)和造模后高剂量CsA治疗组(n=7;造模后1周皮下注射8μg·g-1·d-1 CsA).8周后处死动物,采用免疫组织化学法、逆转录-聚合酶链反应和Western blot检测肾脏MMP-2、MMP-9 mRNA和蛋白表达水平.采用单因素方差分析和直线相关分析进行统计学分析.结果 8周时,单纯糖尿病组24 h尿微量蛋白显著高于正常对照组[分别为(5.80±3.23)、(1.24±0.21)mg/24 h,F=4.229,P＜0.01];各CsA治疗组24 h尿微量蛋白不同程度降低,与正常对照组比较差异无统计学意义.单纯糖尿病组肾小管上皮细胞和肾小球系膜基质MMP-2、MMP-9mRNA(分别为2.22±0.08、2.55±1.10)和蛋白表达(分别为3.1±1.5、2.8±1.0)显著高于正常对照组(MMP-2 mRNA:0.70±0.26,F=6.031;MMP-9 mRNA:0.37±0.24,F=5.193;MMP-2蛋白:1.0±0.0,F=7.532;MMP-9蛋白:1.0±0.0,F=6.100;均P＜0.01).胰岛素治疗对其表达无影响,但CsA干预可下调MMP-2、MMP-9 mRNA和蛋白的异常表达.MMP-2 mRNA与蛋白表达呈显著正相关(r=0.618,P＜0.01),而MMP-9 mRNA与蛋白表达未见相关性(r=0.420,P＞0.05).结论 CsA可减少糖尿病大鼠肾脏MMP-2和MMP-9基因转录和蛋白表达,改善细胞外基质代谢紊乱,可能具有延缓糖尿病肾病发生的作用.     

神经生长因子及其酪氨酸激酶A受体在慢性阻塞性肺疾病大鼠肺泡巨噬细胞中的表达

目的 观察神经生长因子(NGF)及其酪氨酸激酶A(TrkA)受体在COPD大鼠肺泡巨噬细胞的表达变化.方法 40只雄性SD大鼠,随机分为对照组和COPD组,每组20只.采用6个月单纯烟熏法建立COPD大鼠模型,并检测肺功能,HE染色观察肺组织病理变化.从支气管肺泡灌洗液中分离纯化出肺泡巨噬细胞,用酶联免疫吸附法测定肺泡巨噬细胞培养液的上清液中NGF蛋白表达,激光共聚焦法鉴定肺泡巨噬细胞以及半定量检测肺泡巨噬细胞的TrkA蛋白表达.实时荧光定量PCR法检测肺泡巨噬细胞的NGF mRNA及TrkA mRNA表达.两组间比较采用单因素方差分析,组间比较采用t检验.结果 COPD组大鼠肺顺应性为(0.15 ±0.03) ml/cm H2O(1 cm H2O =0.098kPa),每分通气量为(0.045±0.004)L,均显著低于对照组的(0.42±0.05) ml/cm H2O和(0.102±0.010)L,差异均有统计学意义(t值分别为9.46和12.99,均P＜0.01)；气道阻力为(0.038±0.004)cm H20·L-1·S-1,显著高于对照组的(0.016±0.002)cm H2O·L-1·s-1,差异有统计学意义(t=11.55,P＜0.01).肺组织病理显示,COPD组大鼠肺泡壁变薄,肺泡间隔破裂,肺泡腔不规则扩大,部分融合形成肺大疱,肺气肿形成.COPD组NGF蛋白表达量为(3.79±1.52) ng/L,显著高于对照组的(0.94±0.27) ng/L,差异有统计学意义(t=4.13,P ＜0.05).COPD组TrkA蛋白平均荧光强度值(19.5±1.5)显著高于对照组(11.2±1.9),差异有统计学意义(t=7.95,P＜0.05).COPD组NGF和TrkA的mRNA表达量(24.8±6.0和9.0±3.3)显著高于对照组(1.0±0.2和1.0±0.4),差异均有统计学意义(t值分别为8.48和5.16,均P＜0.05).结论 COPD组肺泡巨噬细胞高表达NGF及其TrkA受体,提示NGF及其TrkA受体可能介导肺泡巨噬细胞参与COPD的发病过程.     

爱罗咳喘宁对慢性阻塞性肺疾病大鼠肿瘤坏死因子-α、巨噬细胞炎性蛋白-2、髓过氧化物酶、炎细胞及肺组织病理变化的影响

目的观察爱罗咳喘宁口服液对慢性阻塞性肺疾病(COPD)大鼠模型支气管肺泡灌洗液(BALF)和肺匀浆中肿瘤坏死因子(TNF)-α、巨噬细胞炎性蛋白(MIP)-2、和髓过氧化物酶(MPO)变化及肺组织结构的影响,探讨爱罗咳喘宁方对COPD作用的可能机制。方法采用脂多糖(LPS)加烟雾诱导COPD大鼠模型,大鼠被随机被分为正常组、模型组、爱罗咳喘宁口服液低、中、高剂量组。正常组、模型组灌胃蒸馏水(10 g·kg-1·d-1),爱罗咳喘宁口服液低、中、高剂量组分别灌胃混悬液5,10,20 g·kg-1·d-1,连续7 d。酶联免疫法(ELISA)测定各组大鼠BALF和肺组织匀浆中TNF-α和MIP-2的含量,比色法测定MPO活性。结果与正常组比较,模型组BALF和肺组织匀浆TNF-α、MIP-2和MPO均显著升高(P均<0.05);与模型组相比,爱罗咳喘宁口服液中剂量组TNF-α、MIP-2和MPO均显著降低(P均<0.05)。结论爱罗咳喘宁口服液治疗COPD的作用机制可能与抑制TNF-α、MIP-2和MPO的释放有关。     

As2O3对兔Vx2软组织肿瘤VEGF表达和肿瘤细胞凋亡的影响

目的 探讨不同剂量As2O3对Vx2软组织肿瘤细胞凋亡及血管内皮细胞生长因子(VEGF)表达的影响及As2O3抗肿瘤的作用机制.方法 32只大白兔皮下软组织内肿瘤种植2 w后,随机分为实验组和对照组.实验组根据As2O3用药量不同分为1、2、4 mg·kg-1·d-1 3个亚组,每组8只,对照组注射生理盐水,连续给药7 d,停药后继续观察3 w后处死大白兔,采用MRI测量治疗前后肿瘤最大直径,分别于给药前及处死前行肝肾功能检查,检测As2O3对肝肾毒性作用;采用原位末端标记法(TUNEL法)检测肿瘤细胞的凋亡指数及VEGF染色法观测As2O3抑制肿瘤细胞VEGF表达的程度.结果 MRI显示对照组肿瘤随观察时间延长呈持续增大;实验组随着用药剂量增加,肿瘤增长缓慢(与对照组相比P＜0.05);1、2 mg·kg-1·d-1亚组与4 mg·kg-1·d-1亚组组间差异较明显(P＜0.05),1 mg·kg-1·d-1与2 mg·kg-1·d-1亚组组间差异不明显(P＞0.05).给药前及处死前行耳缘静脉取血,测定ALT、AST、BUN、Cr水平,给药前各组间差异不明显(P＞0.05),处死前基本恢复正常,组间差异不显著(P＞0.05).HE染色显示对照组肿瘤周围组织浸润明显,实验组周围组织浸润不明显;TUNEL法对照组肿瘤坏死周围细胞凋亡与治疗组比较差异显著(P＜0.05),1、2 mg·kg-1·d-1与4 mg·kg-1·d-1亚组组间差异较明显(P＜0.05),1 mg·kg-1·d-1与2 mg·kg-1·d-1亚组组间差异不明显(P＞0.05).VEGF染色法显示肿瘤细胞内VEGF表达随药物浓度增加而减少,与对照组相比差异显著(P＜0.05).结论 经腹腔注射As2O3能够控制Vx2软组织肿瘤增长,同时诱导肿瘤细胞凋亡及抑制肿瘤血管生成,并与剂量相关,其对肝肾功能毒性作用不明显.     

爱罗咳喘宁对慢性阻塞性肺疾病大鼠可溶性细胞间黏附分子1、肿瘤坏死因子-α、白细胞介素-6及炎细胞的影响

目的观察爱罗咳喘宁对慢性阻塞性肺疾病(COPD)大鼠模型血浆及支气管肺泡灌洗液(BALF)中可溶性细胞间黏附分子(s ICAM)-1、肿瘤坏死因子(TNF)-α和白细胞介素(IL)-6含量及炎细胞的影响。方法脂多糖(LPS)加烟雾诱导COPD大鼠随机分为正常组、模型组、爱罗咳喘宁低、中、高剂量组。正常组、模型组灌胃蒸馏水(10 ml·kg-1·d-1),爱罗咳喘宁口服液低、中、高剂量组分别灌胃混悬液(5,10,20 g·kg-1·d-1),连续7 d。酶联免疫法(ELISA)测定各组大鼠血浆和BALF中s ICAM-1、TNF-α和IL-6的含量。结果与正常组比较,模型组血浆和BALF中s ICAM-1、TNF-α和IL-6含量均显著升高(P均0.05);与模型组相比,爱罗咳喘宁中剂量组s ICAM-1、TNF-α和IL-6含量均显著降低(均P0.05)。模型组BALF中炎细胞总数和中性粒细胞数均显著高于正常组(P均0.01);爱罗咳喘宁低、中、高剂量组BALF中白细胞计数、中性粒细胞和巨噬细胞数显著低于模型组(P0.05)。结论爱罗咳喘宁口服液治疗COPD的作用机制可能与抑制s ICAM-1、TNF-α和IL-6释放有关。     

银丹心脑通软胶囊对大鼠急性心肌梗死后Bcl-2、Bax水平的影响

目的探讨银丹心脑通软胶囊对急性心肌梗死(AMI)大鼠Bcl-2、Bax水平的影响。方法结扎冠脉前降支建立AMI模型,将造模成功大鼠随机分为模型组、银丹心脑通大剂量组(1.6 g·kg-1·d-1)、银丹心脑通小剂量组(0.8 g·kg-1·d-1)、阳性药对照卡托普利组(5 mg·kg-1·d-1)各12只,假手术组10只。术后第2天开始灌胃给药,模型组与假手术组给予同等剂量生理盐水,连续4 w。RT-PCR检测非梗死区心肌Bcl-2、Bax mRNA的表达,TUNEL法计算心肌细胞凋亡指数。结果与模型组比较,各给药组的凋亡指数及左室梗死范围明显降低(P<0.05),左心室非梗死区Bax mRNA的表达明显减少(P<0.05),Bcl-2 mRNA的表达明显增加(P<0.05)。结论银丹心脑通软胶囊能够降低AMI后心肌细胞凋亡指数,减少梗死面积,减少左心室非梗死区Bax mRNA的表达,上调Bcl-2 mRNA的表达,提示银丹心脑通软胶囊可通过抑制凋亡通路Bax/Bcl-2改善梗死后心肌细胞凋亡。     

慢性阻塞性肺疾病患者诱导痰中白介素8、白三烯B4、髓过氧化物酶的变化以及噻托溴铵对其的影响

目的 探讨慢性阻塞性肺疾病(COPD)患者中自三烯B4(LTB4)、白介素8(IL-8)、髓过氧化物酶(MPO)的变化及噻托溴铵对其的影响,进一步验证噻托溴铵是否有抗炎作用.方法 选中度至极重度COPD稳定期患者30例为噻托溴铵治疗前组,予吸入噻托溴铵30 d,剂量18μg·d-1,为噻托溴铵治疗后组.并选10例健康查体者为对照组,本组无任何干预措施.三组均进行血气分析、肺功能检查和诱导痰沉渣细胞计数和痰上清液ELISA法测定IL-8、LTB4、MPO.采用独立样本t检验及配对t检验,Pearson及Spearman相关等分析.结果 噻托溴铵组与健康对照组痰细胞分类、IL-8[(777.05±392.43)pg·ml-1vs(311.27±135.93)pg·ml-1]、LTB4[(96.70±29.13)pg·ml-1vs(20.60±9.89)pg·ml-1]、MPO[(23.50±16.77)U·ml-1 vs(10.52±3.55)U·ml-1]指标比较,差异均具有统计学意义.噻托澳铵治疗前后痰细胞分类比较,差异无统计学意义,IL-8[(777.05±392.43)pg·ml-1vs(408.93±339.14)pg·ml-1]、LTB4[(96.70±29.13)pg·ml-1 vs(43.69±16.49)pg·ml-1]差异有统计学意义.相关分析示IL-8与PaO2(r=-0.411)、肺功能指标FEV1%pred(r=-0.440)、FVC%pred(r=-0.440)、FEV1/FVC%(r=-0.417)有相关性.结论 COPD患者诱导痰中炎性细胞及炎性因子与健康对照组比较,差异有统计学意义.噻托溴铵对COPD患者诱导痰中IL-8、LTB4有降低作用.     

多重耐药铜绿假单胞菌耐药状况分析

目的 了解医院铜绿假单胞菌对常用抗菌药物的耐药谱动态变化,为临床合理选用抗菌药物提供依据.方法 应用回顾性调查方法,对本院从2005年1月至2009年6月间临床样本分离的铜绿假单胞菌的药敏试验进行对比统计分析.结果 所分离出的1125株铜绿假单胞菌对常用抗菌药物的耐药率总体呈上升趋势.耐药率最低的是美罗培南（33%）.铜绿假单胞菌对常见抗菌药物的耐药性均有显著性变化.结论 本地区铜绿假单胞菌耐药率有显著性变化,多重耐药现象严重,动态监测其耐药谱变化是防止铜绿假单胞菌的感染率攀升的重要措施.     

冠心病运动处方的最新研究进展

冠心病是目前中国成人心脏病住院和死亡第一位原因,其发病率和病死率呈逐年上升趋势。随着医疗条件的发展,心脏康复知识的普及,冠心病患者康复意识逐渐提高,人们也越来越重视运动锻炼。     

糖尿病对肾移植预后影响的研究

目的探讨肾移植患者合并糖尿病对预后的影响。方法回顾性分析1988年4月至2008年3月中国人民解放军总医院第二附属医院收治的507例肾移植患者手术前后相关临床资料和随访资料。结果507例共行539例次肾移植术(多次手术29例),其中52例合并糖尿病[其中男30例,年龄(51.4±7.1)岁;女22例,年龄(51.6±4.9)岁],移植前糖尿病16例,移植后糖尿病36例;因糖尿病肾病致肾衰竭性肾移植治疗占糖尿病的30.8%。未合并糖尿病肾移植455例[其中男293例,年龄(38.7±12.3)岁;女126例,年龄(43.5±10.1)岁]。合并糖尿病组的各种并发症患病率均显著高于非糖尿病组。合并糖尿病组病死率17.3%(9/52),同期非糖尿病肾移植组病死率7.7%(35/455),尤以移植前糖尿病病死率最高。结论肾移植合并糖尿病者病死率高、合并症多,需加强对肾移植前后糖尿病的控制。     

Influence of Repeated Measurement on One Occasion,on Successive Days,and on Workdays on Home Blood Pressure Values

Few reports provide clear guidelines on how home blood pressure (HBP) should be measured in practice. In this study, we evaluated the influence of repeated HBP measurements on one occasion, and the difference between the 1st and 2nd day and between workdays and nonworkdays. The subjects (468 male, 232 female; mean age 41 years) were recruited from one company. HBP was measured with a semiautomatic device (Omron HEM-759P). Subjects were instructed to perform triplicate morning (m) and evening (e) measurements on 7 consecutive days. HBP tended to decrease during repeated measurements: systolic blood pressure (SBP) was significantly higher the 1st time than the 2nd time and 3rd time. There was no difference in diastolic blood pressure (DBP) between the 1st and 2nd time, but the value the 3rd time was significantly lower than the 1st and 2nd time. Both mHBP and eHBP on the 1st day were significantly higher than those on the 2nd day. mHBP was higher on the 1st workday than on nonworkdays, but the difference was less than 1 mmHg and there was no significant difference. Since there were significant differences in HBP during repeated measurements and between the 1st and 2nd day, which value to adopt as HBP needs to be discussed. Whether HBP was measured on a workday or a nonworkday seemed to have little influence on the HBP values obtained.     

Effects on the diet on brain neurotransmitters.

The synthesis of neurotransmitters in mammalian brain responds rapidly to changes in precursor availability. Serotonin synthesis depends largely on the brain concentrations of L-tryptophan, its precursor amino aicd. This relationship appears to be physiologic: when brain tryptophan levels vary because of insulin secretion or meal ingestion, corresponding alterations occur in the rate of serotonin formation. The ability of any food to modify brain tryptophan (and serotonin) depends on how its ingestion changes the serum concentration of not only tryptophan, but also several other large neutral amino acids that compete with tryptophan for uptake into the brain. Such precursor-induced changes in brain serotonin appear to be functionally important: animals having a reduced level of brain serotonin (caused by the chronic ingestion of a naturally tryptophan-poor diet, such as corn) demonstrate a heightened sensitivity to painful stimuli; this pain sensitivity can be acutely restored to normal values by a single injection of L-tryptophan, which rapidly elevates brain serotonin. The synthesis of catecholamines (e.g., dopamine, norepinephrine) in the brain also varies with the availability of the precursor amino acid L-tyrosine. Single injections of this amino acid increase brain tyrosine levels and accelerate brain catechol synthesis, while injections of a competing neutral amino acid (e.g., leucine, tryptophan) reduce brain tyrosine and its rate of conversion to dopa. The rate of catecholamine synthesis, however, appears to be influenced less by precursor levels than is serotonin formation: tyrosine hydroxylase, whcih catalyzes the rate-limiting step in catecholamine synthesis, responds strongly to end-product inhibition and to other controls that reflect variations in neuronal activity. The synthesis of acetylcholine in brain responds to substrate (choline) availability much like serotonin synthesis. Short-term alterations in brain choline levels are mirrored by similar changes in brain acetylcholine concentration. Variations in the daily dietary intake of choline also modify brain choline and acetylcholine. The relationship between choline availability and acetylchyoline synthesis has already foudn a cletween choline availability and acetylchyoline synthesis has already found a clinical application: choline has been used successfully in the treatment of tardive dyskinesia, a disorder of the central nervous system thought to reflect a deficiency in cholinergic transmission. These relationships between precursor availability from the periphery and brain neurotransmitter synthesis may ultimately provide the brain with information about peripheral metabolic state.