经纤维支气管肺泡灌洗抗真菌药治疗肺部真菌感染临床分析

目的探讨经纤维支气管镜支气管肺泡灌洗抗真菌药治疗肺部真菌感染的临床疗效。方法选取本院收治的80例肺部真菌感染患者,随机将其分为观察组和对照组,各40例,给予对照组患者常规抗真菌药物及对症治疗,观察组患者则在此基础上加用经纤维支气管镜支气管肺泡灌洗抗真菌药治疗。对两组患者的治疗总有效率、不良反应情况及治疗前后动脉血氧分压进行对比。结果观察组患者的治疗总有效率明显高于对照组(P<0.05),两组患者治疗前动脉血氧分压比较差异无统计学意义(P>0.05),治疗后观察组明显优于对照组(P<0.05),两组患者均未发生严重不良反应。结论在常规治疗的基础上给予肺部真菌感染患者经纤维支气管肺泡灌洗抗真菌药治疗可有效地提高治疗效果,且不良反应少,临床效果显著,值得推广和应用。

己酮可可碱对脓毒症致急性肺损伤核因子-κB和明胶酶的干预作用

目的研究己酮可可碱（pentoxifylline，PTX）对急性肺损伤（acutelunginjury，Au）核因子-κB（nuclearfactor-κB，NF-κB）、明胶酶（gelatinase，MMP-2，MMP-9）的干预作用。方法将112只小鼠随机分为盐水对照组（NS组）、己酮可可碱对照组（PTX组）、手术组（CLP组）和己酮可可碱干预组（C＋P组）。采用盲肠结扎穿孔（cecalligationandpuncture，CLP）法复制脓毒症ALI模型，C＋P组在手术基础上尾静脉注射己酮可可碱40mg／kg干预。各组小鼠在对应时间段处死，观察肺病理改变；免疫组化染色检测肺NF-κB染色强度；逆转录-聚合酶链反应（RT-PCR）检测肺NF-κB、MMP-2、MMP-9的mRNA表达；酶联免疫吸附法（ELISA）测定支气管肺泡灌洗液（bronchoalveolarlavagefluid，BALF）中MMP-2、MMP-9的蛋白含量。结果盲肠结扎穿孑L能够引起Au。CLP组ALI病理表现明显，C＋P组较CLP组明显减轻。C＋P组肺组织NF-κB的染色强度、面积低于CLP组（P〈0．05或P〈0．01）。CLP组肺组织NF-κB、MMP-2、MMP-9的mRNA表达以及BALF中的MMP-2、MMP-9蛋白含量均高于C＋P组（P〈0．05）。结论急性肺损伤时，PTX能够抑制NF-κB活化及MMP-2、MMP-9蛋白的过量表达。抑制炎症反应，减轻肺损伤。

Clinical Features and Prognosis in 21 Patients with Extrinsic Allergic Alveolitis

Objective To assess the spectrum of causes, clinical features, differences between disease phases, and prognosis of extrinsic allergic alveolitis （EAA）. Methods Patients with EAA diagnosed at Peking Union Medical College Hospital from August 1983 to May 2007 were analyzed retrospectively. Their medical records were examined to gather clinical, laboratorial, radiological, and histopathological data. Patients were divided to three phases （acute, subacute, and chronic） according to clinical presentations. Follow-up data regarding treatment response, subsequent radio- logical and pulmonary function studies, and clinical outcomes were collected. Results A total of 21 cases were enrolled. Among them, 11 were subacute, 10 were chronic. The most common exposure was pet birds （6 cases, 28.6%）. The primary abnormality of pulmonary function was restriction and/or reduction in diffusing capacity （12 cases, 63.2%）. The most common findings on high-resolution computed tomography （HRCT） were ground-glass opacities （13 cases, 68.4%） and centri- lobular nodules （8 cases, 42.1%）. Airway obstruction in pulmonary function test, emphysema, hmg cysts, and fibrosis on HRCT were more frequently seen in chronic than in subacute patients, though the differences were not statistically significant. Bronchoalveolar lavage fluid （BALF） showed lymphocytosis. The total＇cell count and the percentage of neutrophils were significantly higher in subacute than in chronic patients （P〈0.05）. Nonnecrotizing granulomas were seen in 8 （47.1%） cases. Improvement or normalization in symptoms, radiography, and pulmonary function test after treatment were seen in all 18 patients with avail- able follow-up data. Five patients recurred. Conclusions The characteristic abnormalities of pulmonary function, findings on HRCT, and pa- thology are essential for all phases of EAA, and the atypical manifestations such as obstruction and fibrosis can also be present frequently, particularly in chronic cases. Differential cell counts of BALF are related to the phase of the disease. The treatment response and prognosis of EAA are good.

Lipocalin-2 Test in Distinguishing Acute Lung Injury Cases from Septic Mice Without Acute Lung Injury

Objective To explore whether the amount of lipocalin-2 in the biofluid could reflect the onset of sepsis-induced acute lung injury(ALI) in mice. Methods Lipopolysaccharide(LPS, 10 mg/kg) injection or cecal ligation and puncture(CLP) was performed to induce severe sepsis and ALI in C57 BL/6 male mice randomly divided into 5 groups(n=10 in each group): group A(intraperitoneal LPS injection), group B(intravenous LPS injection via tail vein), group C(CLP with 25% of the cecum ligated), group D(CLP with 75% of the cecum ligated), and the control group(6 sham-operation controls plus 4 saline controls). All the mice received volume resuscitation. Measurements of pulmonary morphological and functional alterations were used to identify the presence of experimental ALI. The expressions of lipocalin-2 and interleukin(IL)-6 in serum, bronchoalveolar lavage fluid(BALF), and lung tissue were quantified at both protein and mRNA levels. The overall abilities of lipocalin-2 and IL-6 tests to diagnose sepsis-induced ALI were evaluated by generating receiver operator characteristic curves(ROC) and computing area under curve(AUC). Results In both group B and group D, most of the "main features" of experimental ALI were reproduced in mice, while group A and group C showed septic syndrome without definite evidence for the presence of ALI. Compared with septic mice without ALI(group A+group C), lipocalin-2 protein expression in septic mice with ALI(group B+group D) was significantly up-regulated in BALF(P<0.01) and in serum(P<0.01), and mRNA expression boosted in lung tissues(all P<0.05). Lipocalin-2 tests performed better than IL-6 tests in recognizing sepsis-induced ALI cases, evidenced by the larger AUC of the former(BALF tests, 0.8800 versus 0.6625; serum tests, 0.8500 versus 0.7000). Using a dual cutoff system to diagnose sepsis-induced ALI, BALF lipocalin-2 test exhibited the highest positive likelihood ratio(13.000) and the lowest negative likelihood ratio(0.077) among the tests of lipocalin-2 and IL-6 in blood and BALF. A statistically significant correlation was found between lipocalin-2 concentration in BALF and that in serum(Spearman r=0.8803,P<0.0001). Conclusions Lipocalin-2 expression is significantly up-regulated in septic ALI mice compared with those without ALI. Lipocalin-2 tests with a dual cutoff system could be an effective tool in distinguishing experimental ALI cases.

Lipopolysaccharide Challenge Induces Long Pentraxin 3 Expression in Mice Independently from Acute Lung Injury

Objective To determine whether the onset of acute lung injury （ALl） induces the up-regulation of pentraxin 3 （PTX3） expression in mice and whether PTX3 concentration in the biofluid can help recognizing sepsis-induced ALI. Methods Wild-type C57BL/6 mice （12-14 weeks old） were randomly divided into 3 groups. Mice in the group 1 （n=12） and group 2 （n=12） were instilled with lipopolysaccharide via intratracheal or intraperitoneal routes, respectively. Mice in the group 3 （n=8） were taken as blank controls. Pulmonary morphological and functional alterations were measured to determine the presence of experimental ALl. PTX3 expression in the lung was quantified at both protein and mRNA levels. PTX3 protein concentration in blood and bronchoalveolar lavage fluid was measured to evaluate its ability to diagnose sepsis-induced ALI by computing area under receiver operator characteristic curve （AUROCC）. Results ALl was commonly confirmed in the group 1 but never in the other groups. PTX3 expression was up-regulated indiscriminately among lipopolysaccharide-challenged mice. PTX3 protein concentration in the biofluid was unable to diagnose sepsis-induced ALl evidenced by its small AUROCC. PTX3 concentration in bronchoalveolar lavage fluid did not correlate with that in serum. Conclusions Lipopolysaccharide challenges induced PTX3 expression in mice regardless of the presence ofALI. PTX3 may act as an indicator of inflammatory response instead of organ injury per se.

Association of swine influenza H1N1 pandemic virus(SIV-H1N1p) with porcine respiratory disease complex in sows from commercial pig farms in Colombia

Porcine respiratory disease complex （PRDC） is a serious health problem that mainly affects growing and finishing pigs. PRDC is caused by a combination of viral and bacterial agents, such as porcine reproductive and respiratory syndrome virus （PRRSV）, swine influenza virus （SIV）, Mycoplasma hyopneumoniae （Myh）, Actinobacillus pleuropneumoniae （APP）, Pasteurella multocida and Porcine circovirus 2 （PCV2）. To characterize the specific role of swine influenza virus in PRDC presentation in Colombia, 11 farms from three major production regions in Colombia were examined in this study. Nasal swabs, bronchial lavage and lung tissue samples were obtained from animals displaying symptoms compatible with SIV. Isolation of SIV was performed in 9-day embryonated chicken eggs or Madin-Darby Canine Kidney （MDCK） cells. Positive isolates, identified via the hemagglutination inhibition test, were further analyzed using PCR. Overall, 7 of the 11 farms were positive for SIV. Notably, sequencing of the gene encoding the hemagglutinin （HA） protein led to grouping of strains into circulating viruses identified during the human outbreak of 2009, classified as pandemic H1N1-2009. Serum samples from 198 gilts and multiparous sows between 2008 and 2009 were obtained to determine antibody presence of APP, Myh, PCV2 and PRRSV in both SIV-H1Nlp-negative and -positive farms, but higher levels were recorded for SIV- HI Nlp-positive farms. Odds ratio （OR） and P values revealed statistically significant differences （p〈0.05） in PRDC presentation in gilts and multiparous sows of farms positive for SIV-HINlp. Our findings indicate that positive farms have increased risk of PRDC presentation, in particular, PCV2, APP and Myh.

Effects of early bronchoalveolar lavage fluid collected from dogs with smoke inhalation injury on the lungs of rats

Objective: Whether early massive bronchoalveolar lavage can remove the harmful substances from the lungs injured with smoke inhalation remains uncertain. This study was designed to observe the effects of early massive bronchoalveolar lavage fluid (BALF) on the healthy lungs in rats. Methods: Mongrel dogs were inflicted with severe smoke inhalation injury. The injured lungs were lavaged with large amount of normal saline in the first hour after injury and the BALF was collected. The BALF was injected into the healthy lungs of 30 rats (group C) in the dosage of 5 ml/kg. The functions and pathological changes of the lungs were observed 24 h after perfusion with the BALF. The data were compared with those of 23 rats (group B) whose lungs were perfused with the BALF collected from normal dogs and those of 21 rats (group A) whose lungs were perfused with normal saline. Results: The mortality rate 24 h after lung perfusion was higher in group C than in groups A and B. The survivors of group C exhibited fluctuation of respiratory rate (RR), remarkable decrease of PaO 2, significantly higher content of lung water, decrease of total static pulmonary compliance and pulmonary expansion index, and increasse of inflammatory cytokines in the tissues of lungs. Only slight mechanic obstructive effect on the airway was observed in rats of group A and B. The pathological changes of the lungs of the rats in group C were similar to those of the dogs with actual smoke inhalation injury. Conclusion: Our findings indicate that the BALF collected from dogs with acute severe smoke inhalation injury in the early stage after injury injured the normal lungs of rats with the bioactive substances in the BALF. These findings show us that it is a valuable therapeutic procedure to apply massive bronchoalveolar fluid lavage in the early stage after inhalation injury.

Analysis of the T cell receptor Vδ region gene repertoire in bronchoalveolar lavage fluid (BALF) and peripheral blood of asthmatics

Objective To explore the role of γδT cells in the airway of asthmatics and to identify the forces which induce and maintain the inflammatory processMethods Peripheral blood (PB) and bronchoalveolar lavage fluid (BALF) were obtained from7 asthmatic subjects and 7 nonsmoker control subjects The percentage of γδT cells in the PB and BALF was measured by immunofluorescent staining and flow cytometry The frequency of usage and the clonality of Vδ subfamilies (Vδ 1-Vδ 3) were assessed by RTPCR and gene scanning Results A higher proportion of γδT cell was detected in the BALF of asthmatic subjects (7.8%±4.7%) than that from control subjects (3.3%±3.0%, P=0.04) No selective usage for a particular Vδ subfamily was found, but the relative expression level of Vδ 1 was significantly higher in the asthmatic airway (44%±13%) than in the control (19%±5%, P=0.0002) In asthmatic subjects, the monoclonal or oligoclonal expansion of γδT lymphocytes was predominant in the BALF, especially Vδ 1+ T lymphocytes Conclusions Antigenic specific γδT cells might play an important role in the inducement and maintenance of airway inflammation Persistent antigenic stimulation may be the key factor that maintains chronic airway inflammation in asthma

Alteration of surfactant proteins A and D in bronchoalveolar lavage fluid of pneumocystis carinii pneumonia

Objective To understand the interaction between surfactant proteins and pneumocystis carinii pneumonia (PCP),and the impact of corticosteriods on surfactant proteins.Methods We established rat models of PCP and bacterial pneumonia induced by subcutaneous injection of 25mg cortisone acetate.At 8- 12 wk,the bronchoalveolar lavage fluid(BALF)of rats was collected.Total nucleated cells of BALF were counted and differentiated,and the concentrations of surfactant protein A(SP-A)and surfactant protein D(SP-D)were measured by immunoblotting assay.The rats were divided into three immunosuppressive groups and a normal control group.Group I,normal control(n = 6),consisted of healthy SD rats;group Ⅱ,negative control(n = 6),consisted of rats with cortisone acetate injection for over 8 wk without lung infection;group Ⅲ,bacterial pneumonia(n = 11),rats were injected with cortisone acetate over 8 wk that resulted in bacterial pneumonia without other pathogens isolated;and group Ⅳ,PCP(n = 14),rats with injected cortisone acetate for 8 - 12 wk and developed PCP without other pathogens isolated.Results Our results indicated that the total cell count in BALF in the negative control group was lower than that in the normal control group(P ＜ 0.001).During PCP infection,the total cell count and the percentage of polymorphonuclearcytes(PMNs)in BALF were significantly increased(P ＜ 0.01),but were lower than those in the bacterial pneumonia group.The concentration of SP-A of BALF in PCP(45.1 ± 22.1 μg/ml)was significantly increased in comparison with that in the negative control(16.2 ± 9.9 μg/ml,P ＜ 0.05)and bacterial pneumonia groups(6.2 ± 5.6 μg/ml,P ＜ 0.001).We also found that the relative content of SP-D was significantly higher in PCP(24249 ±4780 grey values)than that in the negative control (13 384 ± 2887 grey values,P ＜ 0.001)and that in bacterial pneumonia(11 989 ± 2750 grey values,P＜0.001).SP-A and SP-D were also higher in the moderate to heavy group of PCP than those seen in the mild group(P ＜ 0.01,P ＜ 0.001).SP-A and SP-D were higher in the negative control group than those in the normal control group,but there was no significant difference between the 2 groups.Conclusion These results suggest that the concentrations of SP-A and SP-D in BALF are increased by pneumocystis carinii specific stimulation,but the alteration is not related to the corticosteriod usage.

Intervention effect and dose-dependent response of Tanreqing injection on airway inflammation in lipopolysaccharide-induced rats

OBJECTIVE: To assess the effect of Tanreqing injection on airway inflammation in rats. METHODS: A rat model of airway inflammation was generated with lipopolysaccharide (LPS). Tanreqing injection was given by intratracheal instillation, and bronchoalveolar lavage fluid (BALF) from the right lung was collected. BALF total cell and neutrophil counts were then determined. In addition, BALF levels of inflammatory cytokines interleukin-1β, cytokine-induced neutrophil chemoattractant-1, and tumor necrosis factor-α were measured using enzyme linked immunosorbent assay.The middle lobe of the right lung was stained with hematoxylin-eosin and histological changes examined. RESULTS: LPS increased airway inflammation, decreased BALF inflammatory cell count, inflammatory cytokine levels, and suppressed leukocyte influx of the lung. The LPS-induced airway inflammation peaked at 24 h, decreased beginning at 48 h, and had decreased markedly by 96 h. CONCLUSION: Tanreqing injection contains anti-inflammatory properties, and inhibits airway inflammation in a dose-dependent manner.