大鼠吸入贫铀气溶胶后肺组织病理和形态计量学改变

目的:探讨大鼠吸入贫铀(DU)气溶胶后肺组织病理和形态计量学改变,为贫铀武器损伤及其医学防护提供线索。方法:用常规病理技术和形态计量学方法观察了大鼠吸入DU气溶胶后1-14个月肺组织病理和形态计量学改变。结果:吸入DU气溶胶后,部分大鼠发生肺实质内淋巴细胞浸润、严重支气管炎、肺出血、脓肿,且随吸入后时间延长或吸入剂量增高,肺组织病变发生率及病变程度均呈现增高的趋势。结论:大鼠吸入贫铀气溶胶对肺组织有明显的损伤作用。     

贫铀诱发细胞超微结构改变及DMSO的保护作用

目的观察贫铀(DU)对体外培养的人支气管上皮细胞超微结构的影响及二甲基亚砜(DMSO)的保护作用。方法DU作用人支气管上皮细胞(BEAS-2B)24 h后,用荧光染色法检测细胞存活率、坏死率和凋亡率,用透射电子显微镜(TEM)观察细胞超微结构改变。结果荧光显微分析表明,DU作用后,存活细胞数明显减少,凋亡和坏死细胞数显著增高,而DMSO对DU诱发的细胞凋亡和坏死有明显的保护作用。TEM显示,正常细胞和DMSO对照细胞整体形态、核质比例、各种细胞器及细胞骨架均结构清晰;DU处理的细胞,无论细胞内、外有无贫铀颗粒,均见细胞不同程度的凋亡或坏死,正常细胞结构改变或消失,特别是细胞内或外有DU颗粒的细胞,膜性细胞器改变明显,其他细胞器也观察到结构不清或空泡化;DMSO+DU组,即使细胞内外有贫铀分布,各种细胞器结构的改变也明显减轻,观察到有些线粒体、内质网等膜性结构发生膨胀,但细胞整体结构仍较清晰。结论DU可诱发细胞凋亡和坏死,并导致细胞超微结构改变,DMSO对DU所致细胞损伤有明显的保护效果。     

小鼠吸烟急性中毒后肺组织的形态计量学改变

目的:研究小鼠吸烟急性中毒后肺组织的形态计量改变。方法:应用图像分析技术和形态计量学方法,测量小鼠吸烟急性中毒后肺组织的形态计量学改变。结果:吸烟2组（改进型低自由基香烟）小鼠的存活时间是吸烟1组（普通香烟）小鼠的3.7倍。吸烟组与对照组比,小鼠肺泡体积密度、异形指数、等效直径及肺泡间隔厚度明显增大（P<0.05～P<0.01）表面积密度、数密度、形状因子则明显减少（P<0.01）;肺小支气管内腔的形状因子明显增大（P<0.01）,上皮的周长指数显著减少（P<0.01）;小支气管的Reid指数则无明显改变;病理结果显示,吸烟2组病变不如吸烟1组明显。结论:改进型低自由基香烟的毒性明显低于普通香烟。     

氟中毒肾损伤的图像分析研究

目的:观察氟中毒大鼠肾组织损伤的形态定量病理变化特点,探讨过量氟对肾脏的毒性作用。方法:用富钙平衡饲料与低钙偏食饲料分别饲养大鼠,经饮水投氟每升分别含110.5mg、221.0mg、331.5mg氟化钠的高氟水造成水型氟中毒。应用HPIAS—1000电子计算机彩色图像分析系统测试氟中毒大鼠肾组织的鲍曼氏囊腔面积、肾小球周长、血管球周长、肾小管上皮细胞面积、肾小管管腔面积、肾小管管腔面积/肾小管面积之比等。结果:常规食中剂量氟组和高剂量氟组的肾小球球囊面积明显高于常规食对照组(P<0.05),偏食低钙加氟各组肾小球球囊面积也明显高于偏食低钙对照组(P<0.05)。偏食低钙高剂量加氟组肾小管面积较偏食低钙对照组变小(P<0.05),偏食低钙加氟各组较偏食低钙对照组肾小管管腔面积与肾小管上皮细胞比值明显变小(P<0.01)。结论:氟中毒情况下可使肾小球球囊腔增大及其肾小管管腔明显变化,这种损伤性变化可能对肾脏排氟功能有不同程度的影响。     

雾化吸入顺铂对大鼠卵巢功能的损伤情况

目的:了解雾化吸入顺铂对大鼠卵巢功能的损伤情况,同时探讨活性炭口罩是否对顺铂具有防护作用。方法:将60只成年雌性SD大鼠随机分为空白组、活性炭组、普通组及阳性组。空白组仅给予生理盐水雾化吸入,其余各组均给予顺铂2 mg/kg雾化吸入,每天一次,连续2个月。定期检测各组大鼠血清雌激素水平,在实验开始后第15天、30天、60天时取大鼠卵巢组织行病理切片观察。结果:在雾化早期,各组大鼠雌激素水平及卵巢组织病理均无明显变化。雾化60天时,各顺铂雾化组雌激素水平较前明显下降,卵巢组织病理呈现出卵巢体积缩小、卵泡闭锁增多及间质纤维化改变。结论:雾化吸入顺铂对大鼠卵巢功能有一定损害,这种损害随时间延长呈进行性加重。活性炭口罩对雾化顺铂所致卵巢功能损伤防护作用不明显。     

贫铀诱发人支气管上皮细胞恶性转化

背景与目的：实验研究和流行病学调查结果表明,铀可广泛地影响人体健康,但其远期效应,特别是致癌性,还缺乏明确的结论。本文模拟人吸入贫铀（depleted uranium,DU)气溶胶的情形,研究难溶性贫铀诱发人支气管上皮细胞恶性转化及肺癌相关基因表达谱。方法：用难溶性贫铀氧化物（dUO2)作用腺病毒-12/SV40病毒永生化的人支气管上皮细胞（BEAS-2B）,通过观察不同代龄细胞的倍增时间,血清抗性,半固体琼脂克隆形成率及裸鼠成瘤性,鉴定细胞的恶性转化特性;用213个肺癌相关基因的芯片对贫铀诱发的转化BEAS-2B细胞的基因表达谱进行检测。结果：贫铀作用后的第5代BEAS-2B细胞倍增时间明显缩短,血清抗性显著增强;第10代细胞具有锚着独立性生长特性（半固体琼脂克隆形成）;第15代裸鼠体内成瘤。二甲亚砜（DMSO）对贫铀诱发的BEAS-2B细胞恶性转化有明显保护效果。213个肺癌相关基因的芯片检测结果表明,转化细胞中有70多个基因的表达水平发生明显改变,其中10余个基因表达水平明显下降。结论：贫铀在体外具有致癌性。     

氟与砷单独及联合暴露对子代大鼠肾脏毒性损伤与细胞自噬的影响

目的: 评估妊娠期至成年期大鼠慢性氟与砷暴露对肾脏的毒性损伤,并探究潜在的调控机制,为完善氟与砷中毒的发病机制提供科学依据。方法: 试验设置4组即对照组(蒸馏水)、氟暴露组(100 mg/L氟化钠)、砷暴露组(75 mg/L亚砷酸钠)、氟与砷联合暴露组(100 mg/L氟化钠+75 mg/L亚砷酸钠),暴露对象为子代大鼠,从孕鼠妊娠第1天到出生后第21天分别通过母体子宫与母乳的方式暴露于上述氟、砷毒性环境;断乳后,通过饮水染毒至120 d。取子代雄性大鼠新鲜肾脏组织,称取质量后计算肾系数,将肾脏皮质部分通过Masson染色及透射电镜观察肾小球与肾小管的病理改变,并采用免疫荧光技术测定肾小球与肾小管中细胞自噬相关蛋白的表达强度及分布。结果: 与对照组相比,砷暴露组肾系数显著增加(P<0.01);氟、砷单独及联合暴露后肾脏纤维化程度加重,其中氟暴露组肾脏纤维化最明显;此外,所有暴露组中肾小球系膜细胞增生,足细胞内部线粒体空泡化,其中在砷暴露组足细胞中发现典型的自噬小体;相较于对照组,氟、砷单独及联合暴露后肾脏组织中自噬相关蛋白Beclin1、LC3、p62表达水平显著增加(P<0.05),且肾小球与肾小管中自噬相关蛋白的荧光强度与荧光面积也明显增加(P<0.01)。结论: 妊娠期至成年期大鼠氟与砷单独及联合暴露引起肾脏组织中肾小球与肾小管的毒性损伤,并通过增加自噬相关蛋白Beclin1、LC3、p62的表达而调控肾脏细胞自噬的发生。     

大剂量睾酮对雌性小鼠肾损伤的组织学和形态计量学研究

临床上应用雄激素（睾酮）治疗功能性子宫出血，子宫肌瘤和慢性肾病等，用量大，疗程长，且能引起肾功能障碍。为了阐明肾损伤的病理组织学基础，我们给雌性小鼠以大剂量雄激素，用光镜和电镜观察了肾脏的病理组织学和超微病理学改变，并测定了肾小体、血管球和肾小管的形态计量学参数，结果显示：肾小体和肾小管，特别是滤过膜和近曲小管上皮均有十分明显的细胞和亚细胞水平改变；近曲小管的截面积、等效体积、周长、最大直径、最小直径、等效直径和形状因子等７项参数与正常对照组相比，也有十分显著的差异。因此，在长期大剂量应用雄激素时必须重视其对肾脏的损伤     

大鼠急性缺血性肾损伤的细胞死亡方式

目的探讨大鼠缺血再灌注损伤引起急性肾损伤的细胞死亡方式。方法应用光镜和电子显微镜技术、免疫组织化学染色及免疫印迹技术对缺血60min再灌注24h的大鼠肾脏进行了详细观察和分析。结果光镜可见皮质和髓质外带肾小管出现了①大面积细胞坏死：表现为细胞肿胀,空泡形成,崩解脱落;②坏死性凋亡：表现为细胞肿胀,核固缩;③细胞凋亡：分布于髓质外带肾小管坏死细胞之间,表现为细胞皱缩,核固缩。电镜下坏死细胞肿胀,细胞器也肿胀,崩解消失;凋亡细胞胞质皱缩,核染色质固缩边聚;坏死性凋亡呈坏死样细胞质含有一个凋亡样细胞核。免疫组化染色结果显示：PARP-1、RIPk3和caspase-3阳性细胞出现在缺血再灌注损伤肾组织内,主要分布于肾小管。免疫印迹分析结果表明：与Sham组比较肾缺血再灌注后肾组织PARP-1、RIPk3、caspase-3和TNFRa蛋白表达增强（P〈0．05）。结论大鼠肾缺血60min再灌注24h部分肾小管上皮细胞发生了三种方式死亡,即聚合糖性死亡、坏死性凋亡和凋亡。     

肾细胞癌病理诊断中的形态计量学研究

为探讨细胞形态计量学方法在肾细胞癌病理诊断中的价值。方法　采用细胞形态计量学技术对 10 0例肾细胞癌 (RCC)及 10例正常肾组织标本进行测量分析。分析的参数包括 :核面积 ,核形态系数 ,核质比 ,核比轴 ,核短轴 ,轴率等。结果　本研究发现 :1.RCC具有明显的核异型性 ;2 .颗粒细胞癌核面积、核长轴、核短轴均大于透明细胞癌 ;3.核面积、核长轴与核短轴均与核Fuhrman分级呈高度正相关。结论 :细胞形态计量技术 ,完善了传统的单纯依靠经验进行病理诊断的方法 ,具有临床实用价值。     

Oxygen tension in primary gynaecological tumours: the influence of carbon dioxide concentration.

BACKGROUND AND PURPOSE: To assess the effect of inhalation of various high oxygen content gases (HOCG) with different carbon dioxide concentrations on the tumour oxygen tension in patients with primary gynaecological malignancies. MATERIALS AND METHODS: Tumour oxygen tension was assessed on two protocols in those patients with locally advanced visible or palpable primary gynaecological malignancies. Patients were assessed initially while breathing room air (R/A). After 4 min of inhaling the first HOCG, a second assessment of the oxygen tension within the tumour was made. After a 10 min rest period while inhaling R/A, the second HOCG was administered for 4 min after which the third set of measurements were obtained. Protocol A involved assessing the tumour oxygen tension in 12 patients while breathing R/A, 100% oxygen (O(2)) and 5% carbogen (95% O(2), 5% CO(2)). For protocol B, tumour oxygen tension assessments of 13 patients while breathing R/A, 2.5% carbogen (97.5% O(2), 2.5% CO(2)), and 5% carbogen. Median pO(2) and percentage of values 相似文献   

Pulmonary availability of isotretinoin in rats after inhalation of a powder aerosol

Repeated oral administration of chemopreventive retinoids such as isotretinoin over extended periods of time is associated with intolerable systemic toxicity. Here isotretinoin was formulated as a powder aerosol, and its delivery to the lungs of rats was studied with the aim to explore the possibility of minimizing adverse effects associated with its oral administration. Rats received isotretinoin orally (0.5, 1 or 10 mg kg(-1)) or by inhalation (theoretical dose approximately 1 or approximately 10 mg kg(-1)) in a nose-only inhalation chamber. Isotretinoin was quantitated by high-pressure liquid chromatography in plasma and lung tissue. The ratios of mean area of concentration-vs-time curve (AUC) values in the lungs over mean AUCs in the plasma for isotretinoin following single or repeated aerosol exposure surpassed those determined for the oral route by factors of between two (single low-dose) and five (single high-dose). Similarly, the equivalent ratios for the maximal peak concentrations in lungs and plasma obtained after aerosol exposure consistently exceeded those seen after oral administration, suggesting that lungs were exposed to higher isotretinoin concentrations after aerosol inhalation than after oral administration of similar doses. Repeated high doses of isotretinoin by inhalation resulted in moderate loss of body weight, but microscopic investigation of ten tissues including lung and oesophagus did not detect any significant aerosol-induced damage. The results suggest that administration of isotretinoin via powder aerosol inhalation is probably superior to its application via the oral route in terms of achieving efficacious drug concentrations in the lungs.     

Pulmonary carcinogenicity of cadmium by inhalation in animals.

Although, the tumorigenic effect of cadmium after subcutaneous and intramuscular injection has been known since 1961, lung tumours after inhalation exposure were reported only more than 20 years later by Takenaka et al. (1983), using cadmium chloride aerosols. Based on these results, aerosols of cadmium sulfate, cadmium oxide and cadmium sulfide pigment were investigated for carcinogenic effects after inhalation exposure in rats, hamsters and mice. All the cadmium compounds tested appeared to have a very similar carcinogenic potency in the rat lung, whereas no exposure-related tumours occurred in hamsters and in mice only cadmium oxide seemed to increase the lung tumour rate. Subsequent analytical investigations of the aqueous suspension of cadmium sulfide used to generate the aerosol showed that it was soluble in water to some extent as a result of photo-oxidation. The exposure atmosphere therefore actually consisted of a mixture of sulfide and sulfate, and the observed lung tumour rate must be attributed to a combined action of the two compounds. For this reason, a clear-cut carcinogenic effect of cadmium sulfide could not be proved by this inhalation study, but because of the high carcinogenic potency of cadmium ions in the rat lung and because cadmium sulfide retained in the lung is bioavailable to a certain extent, aerosols of this compound should be regarded as a probable human carcinogen.     

组织中氧传输和放射治疗效应影响的初步报道

目的观察放疗时吸入含不同氧浓度的常压气体对放射治疗效价的影响。材料与方法实验肿瘤以Ｔ－７３９小鼠ＬＡ－７９５自发肿瘤在本品系可移植实验肿瘤为观察对象，以再生长延缓为观察指标。正常组织则用评分法对昆明小鼠左右足照射后的皮肤反应程度作为观察指标。对肿瘤及正常组织放射效应的观察均以（１）在空气中照射、（２）先吸入高氧加照射（ＨＯ）、（３）吸入高氧５分钟后再转换为吸低氧（ＬＯ）时同步照射（ＨＯ＋ＬＯ），３个实验组进行相互比较。结果ＨＯ和ＨＯ＋ＬＯ对肿瘤的抑制作用都明显优于空气下照射，两组之间对肿瘤的作用无明显差别，都能达到类似的肿瘤增效作用。ＨＯ明显增加射线对皮肤的损伤，ＨＯ＋ＬＯ虽然也提高了皮肤损伤，但其程度明显比ＨＯ为低。结论放疗时，应用ＨＯ＋ＬＯ放疗方案，在起肿瘤增效和防护正常组织的双重作用方面有潜在可能性。     

The effects of tangential radiotherapy on lung clearance in breast cancer patients.

PURPOSE: The aim of this study was to investigate the effects of tangential radiotherapy (RT) on lung clearance in the patients with breast cancer by using (99m)Tc-DTPA aerosol scintigraphy. MATERIAL AND METHODS: Thirty-three female patients [non-smoker: 20, ex-smoker: 13] performed surgery and systemic chemotherapy for breast carcinoma [47+/-13 years] were included in the study. All patients underwent (99m)Tc-DTPA aerosol scintigraphy prior to RT (pre RT), midway through RT (mid RT) and after RT (post RT). Total dose was 50 Gy in modified radical mastectomy and 60 Gy in lumpectomy (2 Gy/fraction). Posterior dynamic images of lungs were obtained immediately after the inhalation of (99m)Tc-DTPA aerosol. RESULTS: Pulmonary function tests were normal in three measurements for all cases. In the ex-smokers, there was no significant difference among pre RT, mid RT and post RT clearance values in both lungs. Pre RT lung clearance in non-smoker group did not differ from that in ex-smokers. However, the lung clearance for non-smoker group showed significantly increase following RT. CONCLUSION: In this study, we observed that tangential radiotherapy caused an increase in the lung clearance in the cases of non-smokers even in non-irradiated lung, and that the effect of RT on lung clearance was closely depended on smoking habit before RT.     

Distribution of camptothecin after delivery as a liposome aerosol or following intramuscular injection in mice

Purpose: The plant alkaloid camptothecin (CPT) has shown significant antitumor activity against a wide variety of human tumors xenografted in nude mice. In previous studies we have found that administration of dilauroylphosphatidylcholine (DLPC) liposome aerosols containing 9-nitrocamptothecin (9-NC) inhibits the growth of human breast, colon and lung cancer xenografts. The purpose of this study was to analyze the pharmacokinetics and tissue distribution of inhaled CPT formulated in DLPC liposomes. Methods: C57BL/6 mice with subcutaneous Lewis lung carcinoma, Swiss nu/nu mice with human lung carcinoma xenografts and BALB/c mice without tumors were used for pharmacokinetic studies of CPT administered as a liposome aerosol and BALB/c mice were given CPT intramuscularly. Results: After 30 min inhalation of CPT liposome aerosol, drug was deposited in the lungs (310 ng/g) and was followed promptly by the appearance of high concentrations in the liver (192 ng/g) and with lesser amounts appearing in other organs. Drug concentration in the brain was 61 ng/g. After intramuscular injection of CPT dissolved in DMSO, drug was released from the site of injection very slowly and accumulated mainly in the liver (136 ng/g). Only trace amounts appeared in the lungs (2–4 ng/g). These results demonstrate a prompt pulmonary and later systemic distribution of CPT following liposome aerosol administration. Conclusions: The substantial concentrations of CPT in lungs and other organs following inhalation of liposome aerosol suggest the possible benefit of it and of its more active derivative, 9-NC, in the treatment of lung, liver, kidney and brain cancer in humans. Received: 23 September 1998 / Accepted: 11 January 1999     

Aflatoxin B1--DNA adduct formation in rat liver following exposure by aerosol inhalation.

There is growing concern that human exposure to respirable grain dust contaminated with aflatoxin B1 (AFB1), a potent hepatocarcinogen, may be a risk factor for a number of human diseases. The objective of this study was to determine if liver DNA adduct formation occurs in rats following either intratracheal injection or nose-only aerosol inhalation exposure to AFB1. Male Fischer 344 rats were exposed by both routes of administration, and in preliminary data using intratracheal instillation, up to 2% of the administered dose became bound to liver DNA. In the nose-only aerosol inhalation experiments, rats were exposed for up to 120 min. Immediately after exposure, four animals were killed at each time point and their livers removed, DNA isolated and purified and analyzed for aflatoxin-DNA adducts by HPLC. A linear dose-response relationship was observed with a correlation coefficient of 0.96 between increasing length of exposure, and the amount of aflatoxin-N7-guanine adducts formed per mg DNA, the mean values and standard errors were 4.2 +/- 0.18, 15.3 +/- 4.3, 21.6 +/- 2.8 and 56.8 +/- 4.6 pmol aflatoxin-DNA adducts per mg DNA for the 20, 40, 60 and 120 min exposures respectively. The amounts of aflatoxin-DNA adducts formed were statistically significantly different (P less than 0.01) among the treated groups. These results indicate that aerosol inhalation is an effective route of exposure to AFB1 in rats that results in genotoxic damage in the liver.     

Gemcitabine aerosol: in vitro antitumor activity and deposition imaging for preclinical safety assessment in baboons

Aim: To characterize gemcitabine aerosol, its in vitro activity against lung cancer cells, its deposition, and tolerance in a non-human primate model. Methods: In vitro cytotoxicity of nebulized gemcitabine against NCI-H460 and A549 lung cancer cells was tested using a growth inhibition assay and compared with non-nebulized gemcitabine. The ^99mTc-DTPA-radiolabeled gemcitabine aerosol was characterized by cascade impaction and the gemcitabine mass/^99mTc activity relationship was established for further quantitative nuclear imaging. Nine weekly inhalations at a target dose of 1 mg/kg body weight of gemcitabine were performed in three baboons using dynamic scintigraphic acquisitions for continuous monitoring of gemcitabine delivery during inhalation. Gemcitabine plasma concentrations were measured during the first inhalation. Results: Growth inhibition assays for both NCI-H460 and A549 cells did not differ between nebulized and non-nebulized gemcitabine. Aerosol characterization showed a particle mass median aerodynamic diameter of 3.7±0.8 μm and a linear relationship between gemcitabine mass (y) and ^99mTc activity (x) (y=0.82x − 10⁻⁵, R ²=0.88). No toxicity was observed after nine weekly inhalations of a mean dose of gemcitabine of 11.1 mg (88% of the target dose) as assessed from scintigraphic data. A dose-dependent peak plasma concentration of gemcitabine (20–74 ng/ml) was observed by the tenth minute of inhalation. Conclusions: We have characterized a gemcitabine aerosol suitable for intrathoracic airway deposition and demonstrated that jet nebulization does not alter the cytotoxic properties of the drug. In a primate model, we have developed a scintigraphic procedure for the monitoring of aerosol deposition, and we have demonstrated the safety of nine weekly aerosol administrations of gemcitabine.Research supports: This study was supported by a grant from the Comité Départemental du Maine-et-Loire de la Ligue Nationale Contre le Cancer.