血液PEA检测在鉴别PA感染与定植中的作用

目的：探讨大鼠血液铜绿假单胞菌（pseudomonas aeruginosa,PA）外毒素（paeruginosa exotoxin A,PEA）检测在PA感染与定植鉴别中的价值。方法：8周龄雄性SD大鼠180只,经地塞米松抑制免疫预处理后,随机分为感染组（气管内注入0.4 ml铜绿假单胞菌混悬液）、定植组（经咽部滴入铜绿假单胞菌混悬液0.2 ml）、对照组（气管内注入0.4 ml生理盐水）。各组按处死时间不同,再分为3 D组、7 D组、11 D组,分别于接种后第3、7、11 d随机抽取各组20只大鼠,心脏采血法处死,无菌取出肺组织并抽取血样。左肺组织匀浆行铜绿假单胞菌培养并计数菌落;右肺组织HE染色,镜下观察肺组织结构;收集血液分别以实时荧光定量PCR法检测PEA DNA及ELISA法检测IL-6、IL-8、IFN-γ、TNF-α。结果：定植组、对照组大鼠各时间点肺组织均未见PA菌生长及明显炎症反应,定植组各时间点咽拭子培养见PA菌生长（其菌落计数均＞105 cfu/ml）,感染组大鼠肺组织匀浆各时间点均可见PA菌生长（其菌落计数均＞104 cfu/ml）及明显炎症反应,其中以第3 d最为明显,至第7、11天有所下降。各时间点感染组大鼠血液PEA DNA及IL-6、IL-8、IFN-γ、TNF-α表达均有升高,与定植组及对照组表达差异均有统计学意义（P＜0.05）,各时间点定植组及对照组大鼠血液PEA DNA及IL-6、IL-8、IFN-γ、TNF-α表达差异无统计学意义（P〉0.05）,其中感染组大鼠血液PEA DNA及IL-6、IL-8、IFN-γ、TNF-α表达检测以第3 d最为显著。结论：血液PEA DNA检测对鉴别铜绿假单胞菌感染与定植有一定价值,其表达水平高低与大鼠肺组织PA菌菌落计数及肺组织炎症反应程度正相关,推测PEA DNA可能反映肺部感染程度及其预后。     

CD4+ CD25+调节性T细胞在铜绿假单胞菌慢性肺部感染中的变化及其意义

目的 探讨CD4 CD25 调节性T细胞(CD4 CD25 Treg细胞)在铜绿假单胞菌(PA)慢性肺部感染中的变化及意义.方法 60只雄性SD大鼠随机分为PA感染组和空白对照组.将PA包被的硅胶管置人大鼠一侧主支气管建立慢性肺部感染模型,对照组置入无菌硅胶管.28 d后,荧光激活细胞分类(FACS)检测外周血CD4 CD25 Treg细胞数量,ELISA检测血清白细胞介素-10(IL-10)和转化生长因子-β(TGF-β)水平,RT-PCR检测脾脏Foxp3 mRNA表达,肺组织行HE染色观察组织病理学变化.结果 PA感染组Treg/CD4 T淋巴细胞的百分比明显高于对照组[(19.79±6.45)%比(5.15±0.47)%,P＜0.05].PA感染组血清IL-10和TGF-β水平均显著高于对照组[IL-10:(231.52±54.48)pg/mL比(35.43±23.56)pg/mL,P＜0.05;TGF-β:(121.05±7.98)pg/mL比(36.02±8.94)pg/mL,P＜0.05].PA感染组Foxp3 mRNA相对含量较对照组显著升高[(0.80±0.044)比(0.25±0.054),P＜0.05].HE染色见PA感染组肺组织有大量淋巴细胞聚集,纤维增生明显,脓肿形成.结论 CD4 CD25 Treg细胞在PA慢性肺部感染时数量增多、功能增强,对感染慢性化形成有重要影响.     

大鼠铜绿假单胞菌肺部感染TLR4mRNA的水平变化

目的探讨大鼠铜绿假单胞菌（PA）肺部感染与Toll样受体4（rrLR4）mRNA的关系,为临床治疗PA感染提供新的思路。方法采用气管滴注法给予PA生理盐水菌悬液建立PA肺部感染模型。分为对照组和感染PA24、48、72、96h组,分别取大鼠静脉血及肺泡灌洗液进行白细胞计数及分类计数,应用RT—PCR方法检测肺组织匀浆中TLR4mRNA的水平变化。结果PA感染后,大鼠血液白细胞总数于24h时点下降,随后上升并于48h时点达峰值;中性粒细胞与淋巴细胞的比值呈现先上升后下降的变化趋势;肺泡灌洗液白细胞上升,于48h时点达峰值;48h组血白细胞计数,肺泡灌洗液白细胞计数和肺组织匀浆液中TLR4mRNA水平较其他组高。结论PA诱导的肺部感染引起大鼠肺组织中TLR4mRNA含量的改变,这对深入研究PA肺部感染的致病机制有重要意义,为临床提供新的治疗靶点。     

两种铜绿假单孢菌慢性大鼠肺部感染模型的建立方法探讨

目的 建立两种铜绿假单孢菌慢性大鼠肺部感染模型,并根据细菌学和病理学指标对其进行评价.方法 (1)随机将健康清洁级Sprague-Dawley大鼠300只分为3组:野生型铜绿假单孢菌感染组(PAO1组),变异型铜绿假单孢菌感染组(PAO-JP2组)和对照组.(2)应用锐孔喷射装置,将两株铜绿假单孢菌的藻酸盐混悬液制成两种铜绿假单孢菌藻酸盐包裹体凝胶小珠,通过气管插管法将其接种至大鼠肺部,以无菌生理盐水-藻酸盐包裹体接种组作为空白对照组.在接种后3、7、14、28 d,取每组大鼠肺分别进行肺组织匀浆菌落计数和病理学检查.结果 (1)细菌学指标:感染后3、7、14、28 d,对照组肺组织均未培养出细菌,PAO1组和PAO-JP2组肺组织均分别培养出野生型和变异型铜绿假单孢菌,感染后3、7 d,PAO1组和PAO-JP2组大鼠肺组织匀浆菌落计数均＞105 CFU/g(对数值:3 d:PAO1组19±6,PAO-JP2组17±7;7 d:PAO1组13±4,PAO-JP2组:12±4),感染后14、28 d,上述两组大鼠肺组织菌落计数均＞103 CFU/g,PAO1组明显高于PAO-JP2组(P＜0.05)(对数值:14 d:PAO1组11.3±2.8,PAO-JP2组9.6±3.3;28 d:PAO1组9.1±1.5,PAO-JP2组4.2±3.0).(2)病理学指标:接种后3、7 d,感染动物肺组织均有不同程度的脓肿、实变、水肿、出血表现,镜下可见中性粒细胞聚集在藻酸盐包被体周围,形成脓肿.接种后14、28 d,实变逐渐减少,以局部肺不张、纤维增生、肉芽肿形成为主要表现,而以PAO1组表现得更为明显.结论 使用锐孔喷射装置,制成两种铜绿假单孢菌藻酸盐包裹体凝胶小珠并接种至大鼠肺部,可以成功地建立两种细菌的慢性大鼠肺部感染模型,方法较简便可行.     

反复铜绿假单胞菌感染诱导大鼠慢性阻塞性肺疾病模型探讨

目的 探讨反复铜绿假单胞菌(PA)感染能否诱导大鼠产生慢性阻塞性肺疾病(COPD)的病理生理及病理学改变.方法 将120只Wistar大鼠随机分为铜绿假单胞菌感染组(PA组)和对照组(NS组).PA组通过气管穿刺,多次注入一定剂量铜绿假单胞菌菌液,建立大鼠慢性肺部感染模型,测定大鼠动脉血气,观察肺组织病理改变,测量气管壁厚度和血管壁厚度.结果 自感染第4周开始,PA组大鼠体重较NS组显著减轻(p＜0.05);自第12周开始,PA组大鼠血气PaO2显著低于NS组(0.01 ＜P ＜0.05),而PaCO2明显升高(0.01＜P＜0.05);从第2周开始,PA组大鼠气管壁厚度显著大于NS组(P＜0.05);从第4周开始,PA组大鼠血管壁厚度显著大于NS组(P＜0.05);至第16周,PA组大鼠各级支气管慢性炎症明显,管壁增厚,管腔明显狭窄,有气道重构和肺气肿形成.结论 反复气道感染铜绿假单胞菌可致大鼠产生COPD的病理生理及病理学改变,具有COPD的主要特征,提示该方法可能为COPD大鼠模型的建立提供一条新的途径.     

Flt3L对COPD大鼠铜绿假单胞菌肺炎的防治研究

目的 建立铜绿假单胞菌(PA)肺部感染大鼠慢性阻塞性肺疾病(COPD)模型,研究Flt3L(Fms-like tyrosine kinase-3 ligand )对其炎症反应的防治作用.方法 将50只健康雄性清洁级Wistar大鼠随机分为对照组(NS组)、PA感染对照组(NPA组)、COPD大鼠组(COPD组)、COPD大鼠PA感染组(CPA组)和COPD大鼠PA感染Flt3L处理组(Flt3L组),每组动物各10只.用烟雾暴露法复制大鼠COPD模型,气管内滴入PA溶液复制肺部PA感染模型.COPD组、CPA组、Flt3L组大鼠分别在第1～60天以烟雾暴露法复制COPD模型,Flt3L组于第61～65天连续给予Flt3L(20g/kg)腹腔注射,每天1次,连续5d,第66天同时建立NPA组、CPA组和Flt3L组模型.24h后计算大鼠死亡率,对支气管肺泡灌洗液(BALF)、肺组织进行细菌学、细胞学、病理学等研究.无菌操作下取出大鼠脏器进行细菌培养、菌落计数及细菌鉴定.结果 Flt3L组大鼠存活率(87.5%)显著高于CPA组(50.0%)(P<0.01),Flt3L组大鼠支气管肺泡灌洗液中淋巴细胞数[(0.41～0.05)×109/L]与CPA组[(0.24～0.04)×109/L]比较显著增加(P<0.01),光镜下观察比较发现Flt3L组大鼠肺组织炎症反应较CPA组明显减轻,脏器细菌培养阳性率CPA组(37.5%)显著高于Flt3L组(12.5%)(P<0.01).结论 Flt3L能够增强COPD大鼠对PA肺部感染的抗病力.     

sTREM-1在铜绿假单胞菌感染和定植的表达及意义

目的通过研究可溶性髓样细胞触发受体-1(sTREM-1)在铜绿假单胞菌呼吸系统感染和定植不同状态下表达水平的变化,探讨sTREM-1在铜绿假单胞菌肺炎及呼吸道定植中的意义。方法将90只SD大鼠随机分为肺部感染组、口咽部定植组及对照组,每组按铜绿假单胞菌接种后不同时间又分为3、9、24h亚组,每亚组10只。运用酶联免疫吸附法(ELISA)检测血清sTREM-1浓度;采用逆转录-聚合酶链反应(RT-PCR)测定肺组织TREM-1mRNA表达;病理组织切片观察各组肺部炎症改变,并进行肺组织匀浆菌落计数。结果感染组大鼠肺组织病理改变在9h最为严重,而定植组和对照组的肺泡及肺间质结构基本正常;肺部感染组3、9、24h肺组织TREM-1mRNA的表达和血清中sTREM-1的浓度较定植组及对照组均明显增高(P<0.05),定植组与对照组TREM-1mRNA的表达和sTREM-1浓度的比较,差异无统计学意义(P>0.05)。结论检测sTREM-1有助于鉴别铜绿假单胞菌感染与定植;动态监测sTREM-1的变化对感染性疾病的早期诊断和病情评估具有指导意义。     

铜绿假单胞菌肺部感染时局部和血清IL-8动力学变化

目的：观察铜绿假单胞菌肺部感染前后肺局部及血清ＩＬ－８的动态变化，探讨免疫抑制宿主（ＩＣＨ）肺部感染的炎症反应机制。方法：采用环磷酰胺联合氢化的松复制大鼠免疫缺陷模型（抑制组），并以正常大鼠作对照（对照组），经气管注入铜绿假单胞菌复制肺部感染模型，分别于感染后０，３，６，９和２４ｈ留取支气管肺泡灌洗液（ＢＡＬＦ）及血清作细菌 检查并测定其ＩＬ－８浓度（ＥＬＩＳＡ法）。结果：对照组ＢＡＬＦ中ＩＬ－８     

铜绿假单胞菌密度感应系统对气道黏液高分泌的影响

目的 探讨铜绿假单胞菌(PA)的密度感应系统(QS)在气道黏液高分泌中的作用.方法 60只大鼠随机分为3组(野生型菌株PA01组、QS基因突变型菌株PA01-JP2组和对照组),分别将包被不同亚型PA的硅胶管置人大鼠一侧主支气管,建立慢性肺部感染模型,对照组则用生理盐水浸泡硅胶管.28 d后处死大鼠,取肺组织和支气管肺泡灌洗液(BALF)采用AB-PAS染色、ELISA和RT-PCR等方法检测气道内黏蛋白MUC5AC蛋白及mRNA的表达水平,比较野生型菌株PA01和QS突变型菌株PA01-JP2对大鼠气道黏液分泌的影响.结果 PA01组较PA01-JP2组气道黏膜上皮的杯状细胞数目显著增多(P＜0.05),对照组的杯状细胞数目极少.PA01组MUC5ACmRNA表达量较PAO1-JP2组显著增多(P＜0.05),对照组MUC5AC mRNA仅有少量表达.PA01组BALF中MUC5AC蛋白分泌量明显高于PA01-JP2组(P＜0.05).结论 PA的QS重要基因缺损导致气道黏液分泌的能力显著下降.     

铜绿假单胞菌QS系统缺陷对生物膜形成及大鼠肺部慢性感染的影响

目的研究lasI rhlI基因缺陷对铜绿假单胞菌体外生物膜形成的影响;观察铜绿假单胞菌PAO1野生型及其群体感应系统的lasI rhlI基因缺陷型菌株人工生物膜致病性的差异,了解群感应系统的lasI rhlI基因在铜绿假单胞菌生物膜感染致病过程中的作用。方法采用生理盐水-吸痰管系统进行铜绿假单胞菌体外生物膜的培养,3d后用扫描电子显微镜观察PAO1、PAO1 lasI rhlI形成生物膜的情况。从大鼠支气管内直接予以PA(菌种为铜绿假单胞菌PAO1野生型,PAO1 lasI rhlI基因缺陷型)海藻酸盐微菌粒悬液(1×109CFU/mL)攻击,建立PAO1野生型及QS系统的lasI rhlI基因缺陷型菌株人工生物膜肺部感染动物模型。于感染2周后评估各组大鼠的肺部病理学、细菌学的变化。结果3d后PAO1野生型的生物膜较厚,lasI rhlI基因缺陷型菌株所形成的生物膜结构呈薄膜状,明显薄于PAO1野生型。感染2周后,PAO1野生型组的病理学改变和细菌学改变均显著重于lasI rhlI基因缺陷组(P<0.001)。结论QS系统的lasI和rhlI基因影响铜绿假单胞菌体外形成生物膜的能力,并且在PA肺部感染过程中发挥着重要作用。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and ＞ 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.