Unilateral radiation pneumonitis in sheep: physiological changes and bronchoalveolar lavage

Radiation pneumonitis is a life-threatening result of therapeutic thoracic irradiation, yet its mechanisms are poorly understood. We studied the effects of unilateral lung irradiation (3,000 rad) in sheep from the immediate response to the later development of radiation pneumonitis. We defined radiation pneumonitis by its diagnostic clinical feature, radiographic infiltration of the irradiated zone with a straight margin corresponding to the radiation port. The immediate response in the few hours after irradiation was characterized by cough, labored respiration, hypoxemia (arterial PO2 decreased 19 Torr), mild pulmonary hypertension (pulmonary arterial pressure increased 20%), and lymphopenia. Hemodynamics and gas exchange returned to normal by day 2 but became abnormal again before or during radiation pneumonitis at 32 +/- 2 days. Respiratory distress, hypoxemia, and pulmonary hypertension recurred during radiation pneumonitis. Bronchoalveolar lavage during radiation pneumonitis contained increased neutrophils (19 +/- 4%, control = 7%), increased protein (0.27 +/- 0.1 g/dl, control = 0.12 +/- 0.03), and severely impaired ability to lower surface tension. Alveolar macrophages from both lungs during unilateral radiation pneumonitis exhibited impaired generation of superoxide after phorbol myristate (only a 30% increase). Normal control alveolar macrophages increased superoxide production after stimulation greater than 400%. We conclude that unilateral lung irradiation in sheep causes a mild immediate response followed by radiation pneumonitis at 1 mo. Unilateral radiation pneumonitis in this model is associated with ipsilateral neutrophilic alveolitis, increased bronchoalveolar lavage protein, and impaired surfactant function, as well as bilateral functional abnormalities of alveolar macrophages.     

Immunocytochemical observation of paraquat-induced alveolitis with special reference to class II MHC antigens.

The expression of class II major histocompatibility complex (MHC) antigens on alveolar epithelial cells and macrophages was investigated immunocytochemically in paraquat-induced alveolitis in the rat lung. Until 2 days after paraquat injection, class II MHC antigens were expressed on the type II alveolar epithelium without any inflammatory cellular infiltration. From the 4th to the 7th day after paraquat injection, class II MHC antigen-positive macrophages increased in the alveolar spaces, whereas the expression on the type II alveolar epithelium became obscure. Over 10 days after the injection, interstitial fibrosis progressed and the intra-alveolar inflammatory infiltrates decreased. Epithelial cells lining the thickened fibrous septa no longer expressed class II MHC antigens. These results suggest that chemical stimuli can induce class II MHC antigen expression on the type II alveolar epithelium in the early stage of cellular injury, followed by inflammatory infiltration and interstitial fibrosis.     

Immunocytochemical observation of paraquat-induced alveolitis with special reference to class II MHC antigens

The expression of class II major histocompatibility complex (MHC) antigens on alveolar epithelial cells and macrophages was investigated immunocytochemically in paraquat-induced alveolitis in the rat lung. Until 2 days after paraquat injection, class II MHC antigens were expressed on the type II alveolar epithelium without any inflammatory cellular infiltration. From the 4th to the 7th day after paraquat injection, class II MHC antigen-positive macrophages increased in the alveolar spaces, whereas the expression on the type II alveolar epithelium became obscure. Over 10 days after the injection, interstitial fibrosis progressed and the intra-alveolar inflammatory infiltrates decreased. Epithelial cells lining the thickened fibrous septa no longer expressed class II MHC antigens. These results suggest that chemical stimuli can induce class II MHC antigen expression on the type II alveolar epithelium in the early stage of cellular injury, followed by inflammatory infiltration and interstitial fibrosis.     

Influenza virus-induced immune complexes suppress alveolar macrophage phagocytosis

Immune complexes in the lungs are capable of inducing adverse responses. Herein we have detailed the formation of immune complexes in the lungs of influenza virus-infected mice and examined their effect on alveolar macrophage defenses. On days 3, 7, 10, 15, and 30 after aerosol infection with influenza A/PR8/34 virus, the acellular pulmonary lavage fluid was tested for viral antigen, specific viral antibody, and immune complexes by immunoassays. Whereas peak viral antigen (day 3) diminished to undetectable levels by day 10, specific viral antibody remained at a low concentration until day 10, after which it rapidly increased. Immune complex concentrations increased through day 7, peaked at day 10, and gradually returned to the control level by day 30. These data demonstrate that immune complexes of detectable size are induced by influenza virus infection during the interface between antigen excess and antibody excess conditions. Since alveolar macrophages are the pivotal phagocytic defense cells in the lung, the ability of normal alveolar macrophages to ingest opsonized erythrocytes was quantitated in the presence of immune complexes from lavage fluid. Immune complexes from day 10 virus-infected lungs caused a dose-dependent suppression of antibody-mediated phagocytosis to 30% of control values. In contrast, although these immune complexes also markedly decreased the phagocytosis of antibody-coated yeast cells, they did not significantly impair the antibody-independent ingestion of unopsonized yeast cells by macrophages. the suppressive effects of immune complexes on alveolar macrophages may, in part, explain the phagocytic dysfunction that occurs 7 to 10 days after influenza virus pneumonia.     

Protection to glycolysis by a combination of 5-hydroxy-L-tryptophan and 2-aminoethylisothiuronium bromide hydrobromide in lethally irradiated rats.

Rate of glycolysis in vivo at different time intervals following 8 Gy [LD100(30)] whole body gamma radiation (WBGR) was evaluated by estimating liver glycogen, blood sugar, serum lactic dehydrogenase (LDH) and blood lactic acid concentration in adult male Sprague Dawley rats. Within 1 hr of radiation exposure, a significant fall in liver glycogen was observed in rats fed food and water ad libitum. The glycogen content increased after 24 hr and had returned to control level on 7th day after radiation exposure. Blood sugar, serum LDH and blood lactate levels increased significantly as compared to non irradiated controls. Pretreatment with 5-hydroxy-L-tryptophan (5-HTP; 100 mg/kg) + 2-aminoethylisothiuronium bromide hydrobromide (AET; 20 mg/kg) ip 30 min before 8 Gy WBGR, modified these values and restored them to normal level on 7th day post-irradiation.     

Characterization of mononuclear phagocyte subpopulations in the human lung by using monoclonal antibodies: changes in alveolar macrophage phenotype associated with pulmonary sarcoidosis

Current concepts of pulmonary sarcoidosis suggest that the alveolar macrophage plays a central role in the pathogenesis of the disease. To help define the population of alveolar macrophages in sarcoidosis, we compared the surface phenotype of alveolar macrophages from patients with sarcoidosis and from normal individuals by using monoclonal antibodies (63D3, OKM1, M phi P-9, M phi S-1, 61D3, and M phi S-39) that detect surface antigens on cells of monocyte/macrophage lineage. Although almost all blood monocytes expressed surface antigens detected by each of these antibodies, only a minority of normal alveolar macrophages expressed the same surface antigens (p less than 0.05, each comparison). However, in sarcoidosis, the percentage of alveolar macrophages expressing these surface antigens was increased (p less than 0.05, each comparison with normal alveolar macrophages). Several findings supported the conclusion that the increased expression of these monocyte-lineage surface antigens on sarcoid alveolar macrophages resulted from increased recruitment of monocytes to the lung in sarcoidosis and not from abnormal "activation" of alveolar macrophages. First, alveolar macrophages expressing these antigens had an immature morphology. Second, in vitro cultivation of blood monocytes and alveolar macrophages in the presence of immune and inflammatory mediators, including mediators known to be present in the lung in sarcoidosis, did not prevent the loss of expression of monocyte-lineage surface antigens from monocytes or induce reexpression of monocyte-lineage surface antigens on alveolar macrophages. Third, the expression of monocyte-lineage surface antigens was only increased on sarcoid macrophages from patients whose lower respiratory tract contained an increased number of T lymphocytes, cells known to release monocyte chemotactic factor in sarcoidosis. Consistent with the knowledge that corticosteroids usually suppress the alveolitis of active sarcoidosis, when the expression of alveolar macrophage surface antigens was evaluated before and during therapy, the percentage of alveolar macrophages expressing monocyte-lineage surface antigens returned to normal after 1 to 3 mo of therapy.(ABSTRACT TRUNCATED AT 400 WORDS)     

实验性肺纤维化大鼠发病过程中肺组织FIZZ1蛋白和mRNA表达的动态变化

为了探讨FIZZ1（found in inflammatory zone 1）在肺纤维化发病中的作用,应用博莱霉素（5mg／kg体重）气管内注入复制实聆性人鼠肺纤维化模犁,采用HE染色、Masson三联染色、羟脯氨酸含量测定、免疫组织化学染色、原位杂交等方法,观察实验性人鼠肺纤维化的发病过程及其肺组织中FIZZ1蛋白、mRNA表达水平的动态变化。结果显示：（1）实验性人鼠肺纤维化发病过程中,呈现舆型的肺泡炎（7d）、纤维组织增生（14～2ld）及稳定的肺纤维化（28d）等表现;（2）FIZZ1蛋白在正常肺组织表达较弱,存肺纤维化组7d时表达明显增强,14d时较7d时有所减弱,21及28d明显减弱;（3）FIZZ1 mRNA在正常肺组织巾表达较弱,在肺纤维化组7d时表达明显增强,14d时开始减弱,2l及28d明显减弱,但仍强于正常组。上述结果提示,FIZZ1蛋白和mRNA在实验性大鼠肺纤维化发病过程中呈现明显的动态变化,并可能参与了肺纤维化的发生。     

Time course of functional repair of the alveolar epithelium after hyperoxic injury.

The alveolar epithelium is the major barrier to solute and protein flux between the pulmonary vascular bed and the airspaces. Hyperoxic exposure increases epithelial permeability, and during recovery, normal permeability must be regained. To determine the time course for recovery of this function, we exposed hamsters to > 95% O2 for 4.5 days and returned them to room air. After recovery periods of 0.5, 1, 3, 7, and 14 days, alveolar epithelial permeability x surface area (PS) values for [14C]sucrose and fluorescein isothiocyanate-Dextran 20 were measured with isolated perfused lung techniques. Eighty-five percent of the exposed animals survived in room air. Control PS values for sucrose and Dextran 20 were 5.76 x 10(-5) and 0.29 x 10(-5) cm3/s, respectively. After hyperoxia both values were increased by a factor of five. After 0.5 days of recovery, PS remained elevated, but after 1 day they were decreased. Normal PS values were achieved after 3 days for sucrose and 7 days for Dextran 20. During both acute injury and recovery, epithelial selectivity was unchanged and no ultrastructural changes in the alveolar epithelium were observed.     

Changes in spleen histology in response to antigenic stimulation in the snapping turtle,Chelydra serpentine

Proliferative and migratory changes of lymphoid cells in the spleen were observed in turtles stimulated with KLH and maintained at 30°C. Small foci of pyroninophilic lymphoblasts were first seen in the white pulp at about day 5. Progressive enlargement of these centers continued and peaked by days 8-12. By days 15-20 the white pulp returned to a normal (unimmunized) state, while the number of pyroninophilic cells, primarily plasma cells, increased markedly in the red pulp. At days 22-25, the number of plasma cells returned to normal levels and the spleen appeared normal for the remainder of the 60 day observation period. These events suggest that at 30°C,-the turtle is capable of a strong and prompt proliferative response in the white pulp sheaths, followed by migration and differentiation of lymphoblasts into plasma cells, n the red pulp., Observations of pyroninophilic cells in sinuses, venules and veins of the spleen and a concomitant depletion of cells in red pulp, further suggest a migration from the spleen to other sites. Following a second antigenic challenge, at day 60, no significant histological changes were observed at 30°C. Nor were any changes observed following primary or secondary antigenic challenge, in animals maintained at 10°C. These findings are discussed with regard to immunological memory and low temperature immunosuppression in ectothermic vertebrates.     

经鼻滴入博莱霉素致小鼠肺纤维化模型的建立及其病理演变

目的经鼻快速滴入博莱霉素复制小鼠肺纤维化模型,观察肺纤维化模型小鼠的病理形态学改变及其羟脯氨酸含量变化,鉴定肺纤维化模型的成功建立。方法经鼻滴入博莱霉素建立小鼠肺纤维化模型。分别于造模第14天和第28天后,取肺组织行HE染色和天狼猩红染色,取心、肝、脾、肾、脑组织行HE染色,光镜下观察组织病理学变化;造模第28天后用样本碱水解法检测肺组织中羟脯氨酸（HYP）的含量。结果①肺组织病理形态学改变：造模第14天和第28天后模型组小鼠肺泡炎及纤维化程度均明显高于阴性对照组,并且在造模第28天后肺纤维化程度进一步加重;②肺组织中HYP含量：与阴性对照组比较,模型组显著升高（P〈0.05）。结论经鼻滴入博莱霉素可以成功复制小鼠肺纤维化模型,肺纤维化模型小鼠HYP含量升高。     

Maintenance of brain thyroid hormone level during peripheral hypothyroid condition in adult rat

Thyroid hormones are essential for normal functioning of adult mammalian brain. The present investigation deals with the understanding of the time course of thyroid hormone homeostasis in adult rat brain. Animals were rendered hypothyroid by PTU injections (2 mg/100 g bw) for 30 consecutive days. Serum and synaptosomal T3/T4 content, synaptosomal AChE and Na+-K+-ATPase activities were determined on alternate days. While serum T4 level initially increased on the second day compared to control, serum T3 declined in a triphasic pattern; the first phase lasting from the second day to the 6th day, the second phase ended on the 14th day and last phase continued till the 30th day. Cerebro-cortical synaptosomal T3 level increased on the 2nd day from the control, attained a peak on the 4th day, remained stable until the 18th day, and abruptly declined on the 20th day. Synaptosomal T4 content remained negligible or undetected throughout. Synaptosomal membrane Na+-K+-ATPase and AChE activity exhibited an inverse relationship during the experimental regime, being much more prominent on the 2nd, 18th and 20th day coinciding with the variations in brain T3 level. Thus, the study identifies the onset of central homeostasis between the first and second day, its continuation for about 16-18 days and its termination between the 18th and 20th day.     

Experimental Sendai virus infection in laboratory rats. II. Pathology and immunohistochemistry

Intranasal inoculation of 5 to 8 week old specific pathogen-free Sprague-Dawley rats with 5 X 10(3) egg infectious doses of Sendai virus resulted in severe rhinitis, bronchiolitis and alveolitis. The most severe rhinitis occurred on postinoculation (PI) days 4-6, and pneumonia on day 4. Rhinitis and pneumonia persisted to PI day 21, with peribronchial lymphoid infiltration detectable at PI day 42. Immunohistochemical studies showed that Sendai virus antigens were present primarily in columnar epithelial cells of the respiratory mucosa of the nasal cavity and in bronchiolar and alveolar epithelium. Antigen was first detectable at PI day 1, was most prominent at days 3-4 and was undetectable after day 7. More antigen could be seen in the nasal mucosa than in the lung at any stage in the infection. These studies show that Sendai virus by itself is capable of evoking severe, although transient, rhinitis and pneumonia in laboratory rats free of other significant pathogens.     

Food deprivation-induced changes in the level of opioid peptides in the pituitary and brain of rat

Following 1-4 days of food-deprivation (FD) male rats were sacrificed. The pituitary and different regions of brain were analyzed for beta-endorphin-like immunoreactivity (beta-EI), dynorphin (dyn) and methionine-enkephalin (ME) content by RIA. Pituitary beta-EI increased by 16, 28 and 43% on days 2, 3 and 4 of FD. In striatum also, beta-EI increased by 140 and 176% on days 2 and 3 of FD. Dyn level in pituitary was not affected but decreased in hypothalamus by 20% and in striatum by 73% on the 4th day of FD. There was a significant decrease (33-55%) in ME levels in striatum, hippocampus and cortex on 4th day of FD. When food-deprived rats were fed for 24 hr, concentration of most of the opioid peptides returned to basal level. These results suggest that FD in rats affects the opioid peptide levels in a differential manner.     

Effect of exogenous DNA on the content and biosynthesis of endogenous DNA in rat bone marrow]

The influence of exogenous DNA on the content of endogenous DNA and the rate of biosynthesis was studied in rat bone marrow. After the injection of highly-polymerized homologous DNA to intact rats the content of rat DNA per 1 gm of the bone marrow decreased within the first 3 days (the most marked fall occurred in 3 days--by 36%), and returned to the normal by the 6th day. The rate of DNA biosynthesis in rat bone marrow increased in 18 hours (doublled in comparison with control), remained elevated within 6 days (by 58%) and approached the normal level from 1 to 3 days after the DNA injection.     

Damage and recovery of the bone growth mechanism in young rats following 5-fluorouracil acute chemotherapy

Chemotherapy-induced bone growth arrest and osteoporosis are significant problems in paediatric cancer patients, and yet how chemotherapy affects bone growth remains unclear. This study characterised development and resolution of damage caused by acute chemotherapy with antimetabolite 5-fluorouracil (5-FU) in young rats in the growth plate cartilage and metaphyseal bone, two important tissues responsible for bone lengthening. In metaphysis, 5-FU induced apoptosis among osteoblasts and preosteoblasts on days 1-2. In growth plate, chondrocyte apoptosis appeared on days 5-10. Interestingly, Bax was induced prior to apoptosis and Bcl-2 was upregulated during recovery. 5-FU also suppressed cell proliferation on days 1-2. While proliferation returned to normal by day 3 in metaphysis, it recovered partially on day 3, overshot on days 5-7 and normalised by day 10 in growth plate. Histologically, growth plate heights decreased by days 4-5 and returned normal by day 10. In metaphysis, primary spongiosa height was also reduced, mirroring changes in growth plate thickness. In metaphyseal secondary spongiosa, a reduced bone volume was observed on days 7-10 as there were fewer but more separated trabeculae. Starting from day 4, expression of some cartilage/bone matrix proteins and growth factors (TGF-beta1 and IGF-I) was increased. By day 14, cellular activity, histological structure and gene expression had returned normal in both tissues. Therefore, 5-FU chemotherapy affects bone growth directly by inducing apoptosis and inhibiting proliferation at growth plate cartilage and metaphyseal bone; after the acute damage, bone growth mechanism can recover, which is associated with upregulated expression of matrix proteins and growth factors.     

P53抗体联合CYFRA21-1和CEA在非小细胞肺癌中的应用

目的：研究血清P53抗体在非小细胞肺癌临床病理特征之间的关系,并结合血清中的癌胚抗原、角质蛋白21-1以指导对临床上肺癌复发和转移的分析,用来选择合理的治疗方案。方法：正常组30例,肺良性疾病组10例,肺癌组45例,肺癌全组分别于手术前1天、术后10、30、60和90天时抽取清晨空腹静脉血2ml,23例肺癌病例于手术后120天,15例病例于手术后180天抽取清晨空腹静脉血2ml,肺良性疾病组分别于手术前1天、抽取清晨空腹静脉血2ml。正常组清晨空腹采集静脉血2ml。采用酶联免疫吸附法（ELISA）检测血清P53抗体和角质蛋白21-1,采用荧光酶标免疫法检测血清癌胚抗原。结果：血清P53抗体、CYFRA21-1和CEA在正常人组、良性疾病组、肺癌组术前阳性率的比较三种肿瘤标志物阳性率经X2检验,在肺癌组分别与正常人组和良性病例组有显著性差异（P〈0.05）,良性病例组和正常人组之间无显著行差异（P〉0.05）。并与手术后复发与转移相关。结论：联合检测癌胚抗原、角质蛋白21-1及血清P53抗体水平有助于肺部良恶性疾病的诊断;手术前后动态测定肺癌患者血清P53抗体和角质蛋白21-1的变化规律,有助于判断疗效,监测预后和指导肺癌术后的综合治疗。     

Effects of an LHRH agonist on endocrine function, LHRH receptor and LH/hCG receptor in the pituitary-gonadal axis of male rats

The effects of an LHRH agonist (LHRHa), [D-Ser (tBu)]6 des-Gly-NH210) ethylamide, on endocrine function and the LHRH and LH/hCG receptors in the pituitary-gonadal axis were examined. The LHRHa was injected at 100 ng/100 g body weight into male rats once a day for 4 weeks and its effects were observed until 2 weeks after the end of treatment. Due to LHRHa treatment, the plasma LH concentration began to increase on day 3, reached a peak on day 7, and then decreased, although it remained above the control level during the treatment. The pituitary LH content decreased on day 1, reached a minimum (about 40% of the control) between days 3 and 7, and then was maintained at 60% of the control level until week 4. In contrast, the pituitary LHRH receptor concentration increased only on day 3, and the association constant (Ka) remained unchanged during the observation period. The testis weight and plasma testosterone concentration began to decrease on day 3, reached the minimum on day 7 and remained at this level until week 4, and their levels were not completely restored to normal 2 weeks after cessation of treatment. The testicular LH/hCG receptor concentration was decreased on day 1, and markedly decreased to 10-15% of the control value between day 7 and week 4, but the Ka value was slightly increased during the treatment. However, these values had completely recovered 2 weeks after the cessation of treatment. The testicular LHRH receptor concentration increased between days 1 and 7, returned to the control level in weeks 2 and 4, and then decreased 2 weeks after cessation of treatment. Its Ka value was reduced in weeks 2 and 4. These data suggest that the inhibitory effect of LHRHa on the gonad in male rats is not due to reduced pituitary LH release, but to changes in the number and Ka values of gonadal receptors for LH/hCG and LHRH.     

Sulphated glycosaminoglycans in regenerating rat liver.

The total weight percentage glycosaminoglycan content of rat liber was found to increase by 50% in the first 30 h after partial hepatectomy. The content returned to near normal by the third day, but then increased again to a second maximum at 5-6 days, only to gradually decline to normal by the ninth day, when regeneration was nearly complete. This biphasic pattern was most marked in the chondroitin sulphate A/C component, with a 6-fold increase by the sixth day. Dermatan sulphate showed the same temporal trend, whereas heparan sulphate remained relatively unaltered. No such changes were detected in the livers of rats subjected to sham operation. The possible molecular mechanisms underlying the apparent link between cellular glycosaminoglycan content and proliferative tendency are discussed.     

Kinetic parameters of androgen receptor complexes and the activities of the glycolysis and oxidative pentose phosphate pathway key enzymes in rat testis cytosol after whole body 60-min exposure to high frequency electromagnetic field (39.5 Ghz)

The effect of electromagnetic field of millimeter range on biochemical indices of testicles of totally exposed rats at various periods of observation (day 1, 10, 30, 90) was studied. The following functional shifts were revealed: the 6-phosphogluconate dehydrogenase activities enforced approximately up to 130% on 30th day in comparison with the controls, but in three months (90th day) the activity returned to normal values: glucose-6-phosphate dehydrogenase an puruvate kinase did not show obvious changes of activities. On the contrary, there were surprising differences in behavior of androgen-receptor system: the number of androgen-receptor sites in cytosol was increased significantly on 1st day after exposure (114%); on 10th day concentration of RA was quite oppositely 1.68-fold reduced in relation to the control; through 1-month after exposure the rising of [RA] (123%) was registered; however, to 3-month term the significant fall (1.42 times) of the RA contents in testicular cytosol persisted.