缺氧性肺动脉高压大鼠肺动脉一氧化氮合酶的实验研究

目的：研究大鼠肺动脉中一氧化氮合酶（ｎｉｔｒｉｃ ｏｘｉｄｅｓｙｎｔｈａｓｅ,ＮＯＳ）的含量及分布变化与慢性缺氧性肺动脉高压（ｈｙｐｏｘｉｃｐｕｌｍｏｎａｒｙ ｈｙｐｅｒｔｅｎｓｉｏｎ, ＨＰＨ）发生的关系。方法：采用ｅＮＯＳ多克隆抗体,对慢性ＨＰＨ 大鼠的肺组织进行免疫组织化学分析,观察缺氧组及对照组大鼠各级肺动脉中ｅＮＯＳ含量及定位;同时应用分光光度计测定缺氧组及对照组大鼠血浆中ＮＯ的间接浓度。结果：正常大鼠肺动脉内皮细胞及平滑肌细胞均含有较高的ｅＮＯＳ表达,且各级肺动脉中ｅＮＯＳ含量差异无显著性;缺氧组大鼠各级肺动脉内皮细胞及平滑肌细胞ｅＮＯＳ含量均较对照组显著减少;缺氧组大鼠血浆ＮＯ间接浓度较对照组明显降低。结论：大鼠肺动脉内皮细胞和平滑肌细胞ｅＮＯＳ含量下降及血浆ＮＯ浓度下降参与慢性ＨＰＨ 的形成。     

纤维联接蛋白在乙型病毒性肝炎肝组织内的表达

用兔抗人纤维联接蛋白(FN)抗体经PAP法染色,应用形态计量学方法对45例乙型病毒性肝炎患者肝组织内FN的分布进行研究。结果表明,除慢性持续性肝炎组外,各型肝炎组肝小叶内FN 含量均有所减少,其程度与肝炎时肝细胞变性、坏死程度相关。然而慢性持续性肝炎组、慢性活动性肝炎组及亚急性重型肝炎组门管区内FN 含量与正常肝组比较均有所增多。FN 沉积的增多与该区域内纤维结缔组织的增生相平行。     

纤溶异常在慢性阻塞性肺病早期缺氧大鼠肺纤维化中的作用

目的探讨慢性阻塞性肺病（COPD）早期缺氧时纤溶异常与肺纤维化的关系。方法将70只雄性SD大鼠随机分为无缺氧对照组（10只）、COPD缺氧1个月组（一月组,20只）、COPD缺氧2个月组（二月组,20只）及COPD缺氧3个月组（三月组,20只）。模型建立成功后各组大鼠行肺功能测定,处死后肺组织作病理切片,并以HE染色及免疫组化法检测肺组织I、Ⅲ型胶原的表达情况;采用ELISA法测定大鼠肺泡灌洗液（BALF）及血浆中抗凝血酶Ⅲ（AT-Ⅲ）和纤溶酶原激活物抑制剂（PAI-1）的浓度。结果COPD缺氧各组大鼠均形成明显的肺泡炎和肺间质纤维化,并随着缺氧时间延长逐渐加重。与对照组比较,其它三组大鼠I型胶原的表达均明显增强（P〈005）、一月组和二月组Ⅲ型胶原的表达均明显增强（P〈0．05）、三月组Ⅲ型胶原的表达均明显降低（P〈0．05）;与一月组比较。二月组及三月组I型胶原表达均明显增强（P〈0．05）、三月组Ⅲ胶原表达明显降低（P〈0．05）;与二月组比较,三月组I型胶原的表达明显增强（P〈0．05）、Ⅲ型胶原的表达均明显降低（P〈0．05）;三月组I/Ⅲ胶原表达比值较之其它各组均明显增大（P〈0．05）。模型组大鼠BALF及血浆中AT－Ⅲ浓度均较对照组明显降低（均P〈0．05）,二月组及三月组PAI－1浓度均较对照组明显升高（均P〈0.05）;二月组及三月组BALF中PAI－1浓度均较一月组明显升高（均P〈0．05）;三月组BALF中PAI－1浓度较二月组亦明显升高（P〈0.05）。结论COPD早期缺氧所致纤溶异常在肺纤维化的发病过程中起着一定的作用;缺氧时间越长,AT－Ⅲ产生越少,PAI－1合成增多,进而引起肺泡内纤维蛋白形成的增加和清除障碍,这种失衡促进了肺间质纤维化的发生。     

Decreased lung hyaluronan in a model of ARDS in the rat: Effect of an inhibitor of leukocyte elastase

Background.

Hyaluronan (HA) is a component of the extracellular matrix in lung tissue and is normally present at low concentrations in blood. HA is rapidly cleared from blood by the liver. Increased concentrations of plasma HA have been found in patients with acute respiratory distress syndrome (ARDS). We investigated changes in HA levels in plasma, bronchoalveolar lavage fluid (BALF), and lung, and their relationship to pretreatment with a leukocyte elastase inhibitor in a rat model of ARDS.

Methods.

Rats were randomly assigned to three groups: control, thrombin, and thrombin plus elastase inhibitor. By use of a radiometric assay, HA was measured in lungs, BALF, and plasma. Tissue samples from the lungs were stained for HA and examined microscopically. Liver circulation and cardiac output were monitored using radiolabeled microspheres.

Results.

Infusion of thrombin produced a pronounced increase in wet weight to dry weight ratio, and relative lung water content. This increase was blunted by a leukocyte elastase inhibitor. A decrease in lung HA and increases in both BALF and plasma HA were found. The leukocyte elastase inhibitor counteracted not only the decrease in lung tissue HA, but also the increase in plasma HA. Histologically, there was decreased HA-staining of peribronchial and perivascular areas in the injured rat lung. Decreased liver perfusion was observed after infusion of thrombin.

Conclusions.

The decrease in lung HA may be involved in the development of pulmonary edema in this ARDS model, and leukocyte elastase may be one cause of this decrease. In addition, an elevated plasma HA level may be an indicator of lung injury.     

肺泡腔内纤维连接蛋白、透明质酸积聚于博莱霉素致肺损伤早期动态观察

目的：动态观察博莱霉素（ＢＬＭ）致肺纤维化大鼠支气管肺泡灌洗液（ＢＡＬＦ）中纤维连接蛋白（ＦＮ）、透明质酸（ＨＡ）的变化及其与肺内胶原沉积的关系。方法：ＦＮ和ＨＡ测定采用放射免疫法。结果：ＢＡＬＦ中ＦＮ 在早期（第１～７ 天）显著增高，第７ 天达高峰，１４ 天后逐渐下降，至第２８ 天为０；ＢＡＬＦ中ＨＡ 在第３～７ 天明显增高，后逐渐降至正常；ＢＡＬＦ中ＦＮ、ＨＡ 增加先于肺内胶原合成。结论：ＢＡＬＦ中ＦＮ、ＨＡ增高可作为肺泡炎期的标志物     

慢性间断性缺氧及异烟肼、利福平对小鼠肝脏超微结构的影响

目的 探索慢性间断性缺氧（chronic intermittent hypoxia, CIH）及异烟肼（isoniazid,INH,H）、利福平(rifampicin,RIF,R)对小鼠肝脏超微结构的影响。方法 采用两因素两水平的析因分析,检测小鼠肝脏细胞超微结构并对肝脏细胞线粒体形态进行定量分析。结果 慢性间断性缺氧（chronic intermittent hypoxia, CIH）无导致肝脏细胞超微结构损害明显加重,其主效应无导致肝脏细胞线粒体平均截面积、平均周径增加及面数密度减少（P>0.05）;异烟肼（isoniazid,INH,H）、利福平(rifampicin,RIF,R)导致肝脏细胞超微结构损害明显加重,其主效应导致肝脏细胞线粒体平均截面积、平均周径增加及面数密度减少（P<0.05）;两因素两水平的析因分析发现慢性间断性缺氧对异烟肼、利福平引起的肝脏细胞超微结构的损害存在交互作用（P<0.05）。结论CIH协同加重异烟肼、利福平引起的肝脏细胞超微结构的损害。     

Intermittent hypoxia and isoniazid plus rifampicin affect hepatic ultrastructure in mice

Background  Chronic intermittent hypoxia is the most important pathophysiologic feature of sleep apnea syndrome. The present study aimed to determine whether chronic intermittent hypoxia, which is associated with sleep apnea syndrome, can cause or increase damage to liver cell ultrastructure induced by isoniazid and rifampicin in mice.

Methods  Based on a 2×2 full factorial design consisting of two factors of chronic intermittent hypoxia and isoniazid plus rifampicin, 32 male C57B6J mice were randomized into the control group, the chronic intermittent hypoxia group, the isoniazid plus rifampicin group, and the chronic intermittent hypoxia + isoniazid plus rifampicin group. Twelve weeks after treatment, we examined the ultrastructure of liver cells and quantitatively analyzed mitochondrial morphology in C57B6J mice.

Results  Chronic intermittent hypoxia did not significantly affect the ultrastructure of liver cells. The main effect of chronic intermittent hypoxia did not lead to an increase of mean profile area or mean perimeter of mitochondria, and a decrease of numerical density on area of mitochondria (all P >0.05). Isoniazid plus rifampicin significantly affected liver cell ultrastructure. The main effect of isoniazid plus rifampicin resulted in an increase of mean profile area and mean perimeter of mitochondria, and a decrease of numerical density on area of mitochondria (all P <0.05). Moreover, there was a positive interaction among the chronic intermittent hypoxia and the isoniazid plus rifampicin groups for mean profile area, mean perimeter, and numerical density on area of mitochondria (all P <0.05).

Conclusion  Chronic intermittent hypoxia and isoniazid plus rifampicin treatment lead to synergistic liver cell ultrastructural injury.

     

慢性血液透析患者透析前后血浆同型半胱氨酸的变化

OBJECTIVE: To determine the plasma hemocysteine (Hcy) concentration in chronic hemodialysis patients and controls, and the relation between chronic hemodialysis and the plasma total Hcy level in chronic hemodialysis patients. METHODS: Using HPLC-FLD method, the plasma total Hcy concentration was determined in 11 patients with chronic hemodialysis and 30 healthy volunteers. RESULTS: The plasma total Hcy level in chronic hemodialysis was significantly higher than that in the controls (13.54 +/- 4.49 mumol/L) (P < 0.01). The plasma total Hcy level after the hemodialysis (21.30 +/- 10.81 mumol/L) was lower than before the hemodialysis (29.53 +/- 13.59 mumol/L) (P < 0.01). CONCLUSION: The plasma total Hcy level in chronic hemodialysis patients was higher than that in the controls. Hemodialysis can clean the plasma Hcy in chronic hemodialysis patients, but it can not decrease it to the normal level.     

慢性血液透析患者透析前后血浆同型半胱氨酸的变化

目的 :通过对慢性血液透析患者透析前后血浆同型半胱氨酸的检测 ,了解血液透析 (聚砜膜F7透析器 )对慢性血液透析病人血浆同型半胱氨酸水平的影响。方法 :1 1例慢性血液透析患者 ,分别于透析前后 1 0min采取血样 ,另选 30例正常志愿者为对照组 ,抽取清晨空腹静脉血 ,肝素抗凝 ,于 30min内低温离心 7min(70 0g) ,分离血浆 ,采用高效液相色谱仪检测血浆同型半胱氨酸水平。结果 :①慢性血液透析患者血浆同型半胱氨酸水平透前为 (2 9.5 3± 1 3.5 9) μmol/L及透后为 (2 1 .30± 1 0 .81 ) μmol/L ,均明显高于正常对照组 (1 3.5 4±4 .4 9) μmol/L(P <0 .0 1 ) ;②慢性血液透析患者同型半胱氨酸水平透析后较透析前明显降低 (P <0 .0 1 )。结论 :①慢性肾功能不全血液透析患者的血浆同型半胱氨酸平均水平高于正常人 ;②血液透析 (聚砜膜F7透析器 )可以降低慢性肾功能不全血液透析患者血浆同型半胱氨酸的水平 ,对同型半胱氨酸有清除作用 ,但不能使同型半胱氨酸水平降至正常     

急性肺损伤大鼠肺组织内皮素含量的变化及腺苷和黄芪对它的影响

目的：探讨油酸型急性肺损伤（ALI）大鼠内皮素（ET）含量的变化和腺苷、黄芪的保护作用及其机制。方法：观察腺苷、黄芪对ALI大鼠肺组织匀浆ET、支气管肺泡灌洗液（BALF）白蛋白、血清SOD含量及肺湿重、湿/干比、动脉血氧分压（PaO2)变化的影响,并进行组织病理学检查。结果：油酸型ALI大鼠注入腺苷或黄芪能显著降低肺湿重、湿/干比、肺组织匀浆ET和BALF白蛋白含量,提高PaO2和血清SOD水平。结论：腺苷、黄芪能降低油酸型肺损伤的肺水肿和白蛋白渗出,改善缺氧,增强抗氧化,减轻肺损伤程度。     

特发性肺纤维化患者蛋白C系统的研究

目的观察在特发性肺纤维化(IPF)患者支气管肺泡灌洗液(BALF)中凝血酶-抗凝血酶复合物(TAT)和Ⅲ型前胶原(PCⅢ)质量浓度的变化及二者间的相关性。观察IPF患者BALF和血浆中蛋白C(PC)、蛋白S(PS)及血栓调节蛋白(TM)水平的变化,探讨PC系统内各成分质量浓度的变化及其对肺内凝血活性的升高及肺纤维化形成的影响。方法选取正常对照16例,IPF患者16例。IPF患者及8例正常对照行支气管镜检查后取BALF并同时抽血留取血浆,其余8例对照仅留取血浆。采用ELISA法测定BALF及血浆TAT、PC、PS、TM质量浓度及BALF中PCⅢ质量浓度。结果IPF组BALF中PCⅢ和TM质量浓度明显高于对照组(P<0.01,P<0.05),PS质量浓度明显低于对照组(P<0.05)。IPF组血浆中TAT质量浓度明显高于对照组(P<0.01),PC质量浓度明显低于对照组(P<0.05)。IPF组BALF中PCⅢ质量浓度与TAT质量浓度呈线性正相关(r=0.52,P<0.05)。结论IPF患者存在血液的高凝状态。IPF患者肺内凝血活性的升高,与肺内胶原代谢的活动性及肺纤维化的形成相关。IPF患者存在PC及PS质量浓度的下降及TM质量浓度的升高,提示PC途径作用的减弱与凝血活性的升高及肺纤维化的形成具有一定的关系。     

内皮素在低氧性肺动脉高压大鼠肺内的分布及变化研究

为研究低氧性肺动脉高压时大鼠肺内内皮素的分布和变化,以及其与低氧肺动脉高压的关系,用免疫组织化学染色法确定ＥＴ－１在肺内的分布,用族免法测定ＥＴ－１逍度。结果;ＥＴ－１阳性反应部位主要位于内皮细胞,而在内皮与平滑肌连接处浓度更高。缺氧２小时,ＥＴ－１浓度无明显变化,缺氧２４后,ＥＴ－１水平呈升高趋势,并随着缺氧时间的延长而维持在较高水平。     

乳腺良恶性病变中纤维连接蛋白的分布

目的观察乳腺良恶性病变中纤维连接蛋白（ＦＮ）的分布,探讨它与乳腺癌分化、浸润、转移的关系。方法应用免疫组织化学方法,对乳腺良恶性病变纤维连接蛋白进行半定量研究。结果乳腺癌及淋巴结转移细胞ＦＮ呈强阳性表达,而基膜ＦＮ及间质ＦＮ普遍减少甚至缺乏。结论细胞ＦＮ可能是乳腺癌的一个标记物,间质ＦＮ及基膜ＦＮ减少有利于乳腺癌浸润、转移。     

慢性支气管炎大鼠肺一氧化氮和丙二醛产生的变化

[目的 ]测定慢性支气管炎大鼠肺一氧化氮 (NO)、丙二醛 (MDA)含量的变化 ,探讨NO和氧化应激在慢性支气管炎发生发展过程中的作用。 [方法 ]气管内滴细菌脂多糖两次并熏烟 2 8d复制慢性支气管炎动物模型 ,用酶标测定仪检测肺组织匀浆中NO(硝酸盐还原酶法 )、MDA含量与支气管肺泡灌洗液中NO质量分数。 [结果 ]慢性支气管炎大鼠肺组织匀浆NO含量降低于正常大鼠 ,A值分别为 0 .0 5 1和 0 .1 2 6 (P <0 .0 5 ) ,MDA质量分数高于正常大鼠 ,A值分别为 0 .0 2 3和0 .0 1 6 (P <0 .0 5 ) ,而大鼠支气管肺泡灌洗液NO质量分数两组间无统计学差别 ,A值分别为 0 .2 2 5和 0 .2 6 8(P >0 .0 5 )。 [结论 ]一氧化氮、氧化应激在熏烟和细菌脂多糖所致慢性支气管炎发生发展过程中有一定的致病作用     

The effect of pollen in enhancing tolerance to hypoxia and promoting adaptation to highlands

Mixed pollen containing four sorts of pollens (Rape, Typhae, Corn, Sunflower) has been proved to have many biology effects. It is capable of increasing body tolerance to acute hypoxia and promoting adaptation to highlands. The experimental study showed that pollen can significantly increase body tolerance to acute hypoxia pollen can also increase the high energy content and normalize the activity of several enzymes which are important to high energy metabolism; regulate the neurotransmitter in 4 parts of the brain and maintain normal activities in the nervous system; increase the secretion of adrenocortical hormone which may favour O2 absorption; increase SOD content in tissues (heart, liver) and hence may prevent super-oxygenation and guard against free radicals, increase PO2 in the brain and arterial blood; decrease oxygen consumption and blood lactic acid concentration; and increase the immunity of animals under normal condition. Pollen has neither acute nor chronic toxicity and causes no allergic reaction. In field study, pollen can also reduce and ameliorate symptoms of acute mountain sickness in human beings.     

血浆冷沉淀物在深度烧伤创面的实验及临床研究

血浆冷沉淀物是将血浆置于 4℃时 ,获取的一种“冷不溶球蛋白” ,即纤维连接蛋白 (FN)。国内外数次的实验证明 ,FN是体内正常存在的大分子糖蛋白 ,具有很强的生物活性 ,它可作为趋化因子 ,起“净化”伤口的作用 ,为创面愈合创造条件 ;FN也有促进细胞生成的活性 ,促进上皮细胞生长 ,有利于皮片扩展 ;FN还在细胞上皮化过程中起“支架”作用。烧伤后体内FN下降 ,对创面愈合有一定影响 ,而深Ⅱ度烧伤创面基底细胞全部损伤 ,仅依靠残留的汗腺、毛囊上皮扩展修复 ,因而创面修复时间长而缓慢。我们通过动物实验与临床应用 ,观察到 :局部外用富含FN的血浆冷沉淀物 ,可以提高烧伤创面局部FN浓度 ,有利于上皮细胞扩展、移行、加速深Ⅱ度创面上皮化愈合过程。     

新生儿肺出血时肺表面活性物质变化

作者等测定15例新生儿肺出血气道吸出物(TA)和13只兔肺出血模型支气管肺泡灌洗液(BALF)中肺表面活性物质(PS)的生化成分和表面活性。结果发现:肺出血时PS总磷酯(TPL)和主要成分卵磷脂(PC)、磷脂酰甘油(PG)明显减少,总蛋白(TPr)显著增高,表面活性减弱。PS活性减弱的原因可能与TPL、PC、PG的减少及蛋白质对PS的抑制有关。作者提出用外源性PS制剂治疗肺出血,可能会有一定的疗效。     

妊娠肝内胆汁淤积症胎儿宫内缺氧的初步研究

目的　探讨妊娠肝内胆汁淤积症胎儿宫内缺氧与促红细胞生成素 (EPO)之间的关系以及胆酸对胎儿的影响。方法　分别测定于临产前行剖宫产术的正常对照组胎儿 2 5例及ICP组胎儿 32例脐静脉血EPO水平 ;放射免疫法分别测定二组孕妇血清、脐血、羊水胆酸水平 ;胎儿娩出时抽取脐动脉血进行血气分析。结果　ICP组妇女EPO水平同正常对照组相比无差异 ,ICP胎儿脐血EPO及羊水EPO水平均明显低于正常对照组胎儿 ,ICP组缺氧胎儿脐血EPO水平显著低于未发生缺氧的胎儿水平 ,ICP母血、脐血及羊水胆酸与脐血、羊水EPO分别成负相关。结论　ICP胎儿EPO水平低下可能是其宫内缺氧的原因之一 ;ICP妇女血清高胆酸水平可能对胎儿体内合成EPO有一定影响 ;检测羊水EPO或许可以作为产前监测ICP胎儿宫内缺氧的一项指标。     

The preliminary research on the relationship between TNF- and egg-induced granuloma and hepatic fibrosis ofSchistosomiasis Japonica

Summary The amount and distribution of tumor necrosis factor-α (TNF-α) in liver at various stages ofSchistosomiasis japonicum- infected mice were immunohistochemically determined by SABC method. The results showed that the amount of TNF-α in liver began to increase at the 8th–12th week after infection, and reached a peak (9 mice reached 2+ grade or higher) 16 weeks after infection, which was higher than the levels 8 weeks after infection (P = 0. 001) and was mainly scattered in and around egg granuloma. At the chronic stage of the infection, the amount of TNF-α in liver decrease not increase with an increased secretion of collagen in liver and development of hepatic fibrosis. On the contrary, FN, LN, type I and II collagen in liver began to rise 8–12 weeks after infection, and were linearly scattered around egg granuloma, reaching a peak (more than 70 % of infected mice reached 3+–4+ grade) at the 20th and 24th week and they became wide, thick arid retiform, and deposited around and in egg granuloma. After administration of recombinant TNF-α to the infected mice, the amount of FN and LN, the size and the inflammatory response of granuloma in liver showed no significant changes as compared with the untreated group at the same stage. However, the amount of type I collagen in liver increased, among which the amount of type I collagen was significantly higher than that in the untreated group at the same stage (P=0. 01), while similar changes were not observed when recombinant TNF-α was administrated to the healthy mice. It is suggested that TNF may play a role in stimulation of fibroblast in liver to proliferate and secrete collagen, but may need to cooperate with some factors. Supported by the grant from the National Science Foundation of China (94) 330. 2400-240     

肺表面活性物质对哮喘大鼠ET-1及ET-1 mRNA表达的影响

目的:观察吸入肺表面活性物质(PS)对哮喘大鼠模型哮喘发作、内皮素-1(ET-1)及支气管肺组织ET-1 mRNA表达的影响,探讨PS 防治哮喘的机制.方法:建立哮喘大鼠模型,氧气驱动雾化吸入PS,观察哮喘发作情况及肺组织病理变化;采用放射免疫法测定其血浆及支气管肺泡灌洗液(BALF)中ET-1含量 ,以分子原位杂交技术检测支气管肺组织内ET-1 mRNA的表达.结果:哮喘组血浆、BALF中ET-1含量及支气管肺组织ET-1 mRNA表达明显增加;治疗组哮喘发作及病理改变减轻,ET-1含量、支气管肺组织内ET-1 mRNA的表达较哮喘组明显减少.结论:ET-1在哮喘发病机制中可能发挥重要作用,外源性PS能减轻哮喘发作,可能与抑制ET-1 合成及释放有关.