补肺汤对肺纤维化大鼠血清INF-γ IL-4表达水平影响的实验研究

目的：观察补肺汤对肺纤维化大鼠血清IFN-γ、IL-4表达水平的影响,探讨补肺汤对肺纤维化的治疗作用及可能的作用机制。方法：96只SD健康大鼠,随机分为4组（每组各24只）：空白对照组、模型对照组、强的松阳性对照组、补肺汤治疗组。模型对照组和治疗组气管内注射博莱霉素诱导肺纤维化模型,正常对照组在相同条件下给予生理盐水。补肺汤治疗组和强的松阳性对照组分别给予补肺汤流浸膏（2.1g/200g）及强的松片与生理盐水混合制剂（6.3mg/kg）灌胃,其余两组每天给予生理盐水（1mL/100g）灌胃。各组分别于造模后第7、14、28天各处死8只大鼠,取大鼠血清和肺组织。肺组织病理切片HE染色和MASSON染色观察病理变化和肺纤维化分级情况,ELISA法测定血清中INF-γ、IL-4含量的变化。结果：与模型对照组比较,补肺汤治疗组和强的松阳性对照组大鼠各个时期的血清IFN-γ表达明显增高（P〈0.05）;强的松阳性对照组大鼠血清IL-4在各个时期的表达降低明显（P〈0.05）,而补肺汤治疗组在第28天降低明显（P〈0.05）。结论：补肺汤肺纤维化的形成有较好的治疗作用,其机制可能是通过促进血清中IFN-γ的分泌,抑制IL-4的分泌,从而调节了Th1/Th2细胞因子失衡来实现的。     

In vivo MRI of altered proton signal intensity and T2 relaxation in a bleomycin model of pulmonary inflammation and fibrosis

Purpose:

To investigate the ability of proton (¹H) magnetic resonance imaging (MRI) to distinguish between pulmonary inflammation and fibrosis.

Materials and Methods:

Three groups of Sprague‐Dawley rats (n = 5) were instilled intratracheally with bleomycin (2.5 U/kg or 3.5 U/kg) in saline or with saline only. Rats were imaged at 2.0 Tesla using a multi‐slice Carr‐Purcell‐Meilboom‐Gill (CPMG) sequence with 6 ms echo spacing. Signal intensity (S₀) and T₂ were calculated on a pixel‐by‐pixel basis using images collected before dosing and 1, 2, 4, and 7 weeks after. At each time point, data from dosed animals were compared with controls, and bivariate statistical analysis was used to classify image pixels containing abnormal tissue. At week 7, pulmonary function tests were performed, then all rats were killed, left lungs were formalin fixed and tri‐chrome stained for histological analysis of collagen content, and right lungs were used to measure water and hydroxyproline (collagen) content.

Results:

The product S₀×T₂ significantly correlated with water and collagen content in the high‐dose group (P = 0.004 and P = 0.03, respectively). However, S₀ and T₂ of abnormal tissue were correlated for all time points (r = 0.93, P < 0.001), and could not distinguish inflammation from fibrosis.

Conclusion:

MRI can be used to confidently localize pulmonary inflammation and fibrosis, but it lacks specificity. J. Magn. Reson. Imaging 2010;31:1091–1099. © 2010 Wiley‐Liss, Inc.     

Time course of bleomycin-induced lung fibrosis

Intratracheal instillation (IT) of bleomycin is a widely used experimental model for lung fibrosis. In this study we describe the time-course of bleomycin-induced lung fibrosis in mice using computer-assisted morphometry. C57Bl/6J mice were treated with a single IT dose of bleomycin or control saline. Animals were killed 3, 6, 14 and 21 days post-IT. Lung injury was evaluated by analysis of bronchoalveolar lavage (BAL) fluid, hydroxyproline concentration in the lung, routine light microscopic examination resulting in a semiquantitative morphological index (SMI) of lung injury, and quantitative morphological measurements (fibrosis fraction and alveolar wall area fraction) aided by optimas image analysis software. Changes in BAL fluid attributed to bleomycin treatment include increased total cell count (days 14 and 21), and increased percentage of neutrophils (days 3 and 6) followed by a sustained increase in lymphocytes (days 6, 14 and 21). Hydroxyproline levels increased in bleomycin-treated mice on days 14 and 21. Median SMI grades were significantly elevated on days 3, 14 and 21. Computer-assisted morphometry demonstrated a 3-fold increase in fibrosis fraction and a 1.3-fold increase in wall area fraction in bleomycin-treated mice on day 14, with no further increase on day 21. These data also demonstrate that the most suitable time point for assessing lung fibrosis in this model is 14 days after IT instillation of bleomycin, based on the observation that at 14 days the animals developed extensive fibrosis, but had less variability in the fibrotic response and lower mortality than later at 21 days. Computer-assisted morphometry provides objective and quantitative measurements that are a useful tool for the evaluation of bleomycin-induced lung injury.     

RAD6 gene is involved in heat shock induction of bleomycin resistance in Saccharomyces cerevisiae

Cells react to environmental and endogenous challenges such as high temperature, reactive oxygen species, DNA damage, and nutrient starvation by activating several defense mechanisms known as stress responses. An important feature is the overlap between different stress responses that contributes at least in part to the phenomenon of cross-protection. We previously demonstrated that pretreatment with a heat shock (HS) induces resistance to the lethal and mutagenic effects of the antineoplastic drug Bleomycin (BLM) in wild-type Saccharomyces cerevisiae. At the DNA level, the HS resulted in more efficient repair of BLM-induced DNA damage. In the present study, we have investigated the mechanisms involved in this HS-induced BLM resistance. Since the RAD6 gene is involved in the ubiquitin system and DNA repair, we analyzed the effects of HS on the lethality of BLM in a rad6Delta (ubc2) mutant strain of S. cerevisiae. The rad6Delta mutant was more sensitive to the lethal effects of BLM than wild-type yeast and HS had no effect on the lethality of BLM in the mutant. Analysis of cell proliferation kinetics indicated that the HS-induced cell cycle delay observed in the wild-type yeast was absent in the rad6Delta mutant strain. BLM treatment impaired mutant cell proliferation, and HS had no effect on the delayed cell kinetics of the mutant. In addition, pulsed-field electrophoresis of chromosomes damaged by BLM indicated that there was very little recovery from damage in the mutant after 24 hr of incubation in BLM-free nutrient medium, and that HS had little effect on the recovery. These data indicate that the RAD6 gene is involved in the HS-induced BLM resistance observed in the isogenic wild-type strain.     

环孢素A抑制博来霉素诱导的小鼠肺间质病变

目的:观察环孢素A(CsA)对博来霉素(BLM)所致肺间质病变的治疗作用,并探讨其作用机制。方法:将120只C57BL/6小鼠随机分为5组,模型组、模型对照组、CsA30mg治疗组、CsA50 mg治疗组和治疗对照组,每组24只;模型组及治疗组小鼠均经气管滴入BLM建立肺间质病变模型,模型对照组小鼠则经气管滴入同体积生理盐水。于造模当日起,治疗组经腹腔注射CsA,治疗对照组经腹腔注射等体积生理盐水,模型组及模型对照组不予治疗。在实验第4、7和14天分别抽取小鼠外周血,用细胞计数板计数白细胞总数,以流式细胞术检测CD4+T细胞、CD14+单核细胞和CD19+B细胞的数量比例;同步取支气管肺泡灌洗液(BLAF)细胞计数及细胞涂片Giemsa染色分析;并进行肺组织病理学研究。结果:模型组外周血白细胞总数明显升高,细胞分类显示CD14+单核细胞和CD19+B细胞在造模4 d、7 d、14 d均较模型对照组显著增高。CsA治疗后,CD14+细胞比例在4 d时下降最为明显,显著低于同一时间点模型组;7 d、14 d亦低于同一时间点模型组;治疗组CD19+B细胞比例在7 d、14 d时均显著低于同一时间点模型组,各组间CD4+T细胞比较均未见明显统计学差异;BALF细胞分析显示,模型组4 d、7 d、14 d时细胞总数及分类计数均明显高于同一时间点模型对照组,CsA治疗后均显著减少;肺组织病理学观察可见,在第4～14天,模型组及治疗对照组小鼠肺间质内浸润细胞逐渐增多,其中以淋巴细胞浸润为主,可见少量单核-巨噬细胞浸润,同时见间质内胶原纤维沉积逐渐增多。CsA治疗组小鼠肺间质炎症明显减轻,间质内及小支气管周围胶原纤维沉积减少;免疫组化研究显示,模型组小鼠和治疗对照组肺内组织内浸润的单核-巨噬细胞、中性粒细胞和淋巴细胞均高表达CD147分子,治疗组肺内表达CD147的细胞减少。结论:环孢素A可能通过抑制CD147-CypA的相互作用,抑制炎性细胞向肺内的趋化聚集,减轻肺部炎症、减少胶原沉积。     

博来霉素A2和B2的HPLC测定

建立了HPLC法测定博来霉素A_2和B_2。采用C_(18)柱,以硫酸钾缓冲液和乙腈为流动相,梯度洗脱,检测波长为254nm。博来霉素A_2和B_2均在0.05～2mg/ml浓度范围内线性关系良好,平均回收率为99.1%和107.2%,RSD为0.96%和1.58%。     

链霉素抗性筛选法在选育博来霉素A2组分高产菌株中的应用

应用链霉素抗性筛选法提高博来霉素产生菌Streptomycin verticillus SIPI 7011产博来霉素A2组分的产量，将经紫外线诱变处理过的博来霉素产生菌孢子涂布在含有链霉素最小抑制浓度（180μg／ml）的培养基平板上，获得了151株链霉素抗性突变株。其中博来霉素A2产量高于出发菌株的有19株，产量阳性效率达到12．6％，并获得一株产抗生素能力为出发菌株约1．25倍的突变株。     

平阳霉素局部注射治疗睑黄疣的临床疗效观察

目的观察平阳霉素局部注射治疗睑黄疣的临床疗效。方法对87例睑黄疣患者在常规消毒后以平阳霉素8mg加2％利多卡因2mL溶解后行皮损内局部注射治疗作为治疗组，并设立二氧化碳激光治疗45例作为对照组。结果治疗组痊愈74例，显效9例，总有效率95．4％；对照组痊愈16例，显效14例，总有效率66．7％；两组比较差异有统计学意义（U=4．02，P〈0．01）。结论平阳霉素治疗睑黄疣疗效较常规治疗方法好。     

Cytotoxic chemotherapy for patients with terminal squamous carcinoma-does it influence survival?

The absence of any good scientific evidence that a single cytotoxic chemotherapy regimen is effective in the palliation of squamous carcinoma of the head and neck is reflected in the diverse manner with which palliative chemotherapy is used in the UK. We conducted a survey of the members of the Association of Head and Neck Oncologists of Great Britain: 70% of the specialists questioned used palliative chemotherapy for head and neck cancer. Bleomycin was cited by 80% of the chemotherapy users, whereas cisplatin was cited only by 35% We also report the results of a controlled trial of palliative chemotherapy in head and neck cancer. Cisplatin prolonged survival and time spent at home but Bleomycin did not. Also there appeared to be a synergistic effect between Cisplatin and Bleomycin     

抗氧化剂对博莱霉素拮抗作用的研究

目的观察抗氧化剂（维生素E、牛黄酸、二甲亚砜和维生素C）针对博莱霉素（Bleomycin,BLM）的拮抗作用.方法在人外周血淋巴细胞培养物中注入一定剂量的BLM或并一定剂量的抗氧化剂,然后观察淋巴细胞的姐妹染色单体交换率（sister chromatid exchanges ,SCE率）的变化.结果维生素E浓度为5 μl/ml时,与28.6 nmol/ml的BLM处理组（对照2）相比,可显著地拮抗BLM的作用（P〈0.05）;而维生素E浓度为10 μl/ml时,可极显著地拮抗BLM的作用（P〈0.01）.当加入牛黄酸的终浓度为100和200 μg/ml时,与28.6 nmol/ml的BLM处理组（对照2）相比,均可显著地拮抗BLM的作用（P〈0.05）.结论一定剂量的维生素E和牛黄酸对BLM存在着拮抗作用.