Ecografía pulmonar en la evaluación del paciente crítico / Lung ultrasound in the evaluation of critically ill patients

Introducción: La ecografía es una técnica de imagen no invasiva que permite explorar diferentes órganos de manera inmediata, constituye un instrumento de alto valor diagnóstico al alcance del profesional de la salud, y es utilizada en todas las especialidades médicas. En los últimos años, la evolución tecnológica ha permitido que los aparatos de ecografía sean más pequeños, portátiles, y con una alta resolución, tal es el caso de la ecografía clínica o ecografía a pie de cama. La ecografía del paciente crítico ha cambiado la práctica médica; específicamente la ecografía pulmonar se debe realizar en todos los pacientes con enfermedad pulmonar aguda. Objetivo: Aportar el conocimiento teórico necesario para promover el uso de la ecografía pulmonar en la evaluación del paciente crítico, y contribuir, mediante su aplicación, a la disminución del riesgo de exposición a las radiografías. Métodos: Se efectuó una revisión de la literatura médica actualizada sobre el papel de la ecografía pulmonar en la evaluación del paciente crítico, en el período de julio a diciembre de 2021. Se utilizaron los siguientes motores de búsqueda: SciELO, Medigraphic y Google Académico. Conclusiones: En el contexto actual, la ecografía pulmonar ha adquirido un papel protagónico, pues su uso facilita una evaluación frecuente y no invasiva del paciente crítico con afección pleuropulmonar. Su aplicación garantiza la disminución del riesgo de exposición a las radiografías.

Introduction: ultrasound is a non-invasive imaging technique that allows us to explore different organs immediately; it constitutes an instrument of high diagnostic value within the reach of health professionals and used in all medical specialties. In recent years, technological evolution has allowed ultrasound devices to be smaller, portable and with high resolution, such is the case of clinical ultrasound or bedside ultrasound. Bedside ultrasound in critically ill patients has changed medical practice; specifically, lung ultrasound should be performed in all patients with acute lung disease. Objective: to provide the necessary theoretical knowledge in order to promote the use of lung ultrasound in the evaluation of critically ill patients, as well as to contribute, through its application, to reduce the risk of exposure to radiographs. Methods: a review of the updated medical literature on the role of lung ultrasound in the evaluation of the critically ill patients was performed from July to December 2021. SciELO, Medigraphic and Google Scholar were the search engines used. Conclusions: lung ultrasound has acquired a leading role in the current context, since its use facilitates a non-invasive and common evaluation of the criticall ill patients with pleuropulmonary disease. Its application guarantees the reduction of the risk of exposure to X-rays.

Effects of antenatal dexamethasone and postnatal pulmonary surfactant combined with respiratory support on lung fluid clearance in preterm rabbits / 中华围产医学杂志

Objective:To explore the effects of prenatal dexamethasone (DEX), postnatal pulmonary surfactant (PS) and respiratory support on the lung fluid clearance in premature rabbits at gestational age (GA) of 25-28 d (full term: 31 d) and their relationship with dynamic compliance of respiratory system (Cdyn), pulmonary morphology and other parameters.Methods:In our previous publications, premature rabbits were divided into four groups according to the intervention strategy: control group, PS-only group, DEX-only group and DEX+PS group in which data of several parameters including wet-to-dry lung weight ratio (W/D), Cdyn and volume density of alveoli (Vv) were retrieved and the lung tissue sections were scanned to recalculate the ratio of perivascular sheath to vascular sectional area (S/V) and lung injury scores-edema (LIS-E). W/D, LIS-E, S/V and Vv were adjusted for birth weight (BW) (divided by BW, represented as W/D/BW, LIS-E/BW, S/V/BW and Vv/BW) and mean Cdyn (Cdyn-m) was adopted. Based on the grouping of previous studies, the intervention groups in this study were divided as DEX group and non-DEX group, and PS group and non-PS group to analyze the influence of DEX and PS on the above parameters. Two independent samples t-test, one-way analysis of variance, LSD test, Kruskal-Wallis H test, Mann-Whitney U test and Pearson correlation analysis were used for statistical analysis. Results:A total of 196 newborn rabbits receiving mechanical ventilation after birth were included in this study. (1) Effects of DEX: compared with the non-DEX group, the DEX group showed increased W/D/BW (489±69 vs 421±113, t=－2.09), LIS-E/BW (188±57 vs 138±55, t=－2.61) and Vv/BW (20.1±4.9 vs 14.2±4.7, t=－3.60), but decreased S/V (0.33±0.23 vs 0.51±0.25, t=2.23) and S/V/W/D (0.05±0.03 vs 0.07±0.04, t=2.22) at 25 d of gestation; at 26 d of gestation, W/D/BW (472±76 vs 303±44, t=－8.75), LIS-E/BW (189±63 vs 106±36, t=－5.23), Cdyn-m [(0.16±0.07) vs (0.05±0.03) ml/(kg?cmH 2O), 1 cmH 2O=0.098 kPa; t=－7.29] and Vv/BW increased (22.4±5.0 vs 12.2±3.8, t=－7.46), while S/V (0.23±0.19 vs 0.62±0.38, t=4.10), S/V/BW (15.7±12.4 vs 25.7±17.3, t=2.20), S/V/W/D (0.03±0.03 vs 0.08±0.05, t=3.92) and propensity scores decreased [(12.5±1.2) vs (15.1±1.2) scores, t=7.00]; at 27 d of gestation, Cdyn-m increased [(0.23±0.12) vs (0.16±0.07) ml/(kg?cmH 2O), t=－2.43], but S/V (0.32±0.23 vs 0.57±0.39, t=2.57) and S/V/W/D decreased (0.05±0.04 vs 0.09±0.06, t=2.55); at 28 d of gestation, W/D/BW (270±64 vs 162±33, t=－8.09), LIS-E/BW (72±32 vs 35±20, t=－5.17), S/V (0.90±0.60 vs 0.59±0.48, t=－2.81), S/V/BW (34.0±23.6 vs 15.2±12.7, t=－3.77) and Vv/BW increased (16.9±4.3 vs 9.2±2.9, t=－8.04); the differences were all statistically significant (all P<0.05). (2) Effects of PS: compared with the non-PS group, the PS group had decreased LIS-E/BW at 25, 26 and 27 d of gestation, increased Cdyn-m and Vv/BW at 25 and 27 d of gestation and higher propensity scores at 25 d of gestation (all P<0.05). (3) The correlation between gestational age and each index: gestational age was positively correlated with S/V ( r=0.31, P<0.05), but negatively correlated with W/D/BW and LIS-E/BW ( r=－0.73 and－0.63, both P<0.05). Conclusions:The pharmacological action of prenatal DEX on lung fluid clearance is mainly confined to preterm rabbits at the GA of 28 d which is supported by mechanical ventilation. Prenatal treatment with DEX and/or postnatal PS can improve the early respiratory function in preterm rabbits between GA of 25-27 d, but had no substantial impact on lung fluid clearance. The GA-related lung maturation appears to play a crucial role, in comparison with medications, in lung fluid clearance.

Effect of apneic oxygen insufflation on phenotypic transformation of alveolar macrophage in non-ventilated lung during one-lung ventilation / 中华麻醉学杂志

Objective:To evaluate the effect of apneic oxygen insufflation (AOI) on phenotypic transformation of alveolar macrophage (AM) in the non-ventilated lung during one-lung ventilation (OLV).Methods:A total of 60 patients of either sex, aged 40-64 yr, weighing 45-85 kg, undergoing elective thoracoscopic lobectomy, were recruited and divided into 2 groups using a computer-generated table of random numbers: test group and control group, with 30 cases in each group.At the beginning of OLV, the non-ventilated lung received 3 L/min of AOI in test group and no AOI in control group.Radial artery blood samples were collected for blood gas analysis before operation, immediately after anesthesia induction, 30 min, 1 h and 2 h after the start of OLV, and oxygenation index (OI) was calculated.The resected normal lung tissues around the lung lobe were excised at 2 h after the start of OLV for microscopic examination of the pathological changes after HE staining, and the lung injury score was assessed.Bronchoalveolar lavage fluid (BALF) was collected at 2 h after the start of OLV, AM was sorted by flow cytometry, and the apoptotic rate was calculated.The levels of intracellular Ca 2+ and reactive oxygen species (ROS, a marker of M1 AM phenotype) in cells were determined.The concentrations of M1 phenotype AM markers inducible nitric oxide synthase (iNOS), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF-α) and of M2 phenotype AM markers arginase 1 (Arg-1) and interleukin 10 (IL-10) in BALF were measured by enzyme-linked immunosorbent assay. Results:Compared with control group, SpO 2, PaO 2 and OI were significantly increased, PaCO 2 and lung injury score were decreased, the survival rate of AM was increased, the apoptotic rate in the early and late stages was decreased, the concentrations of iNOS, IL-6 and TNF-α in BALF were decreased, and the concentrations of Arg-1 and IL-10 in BALF were increased, the level of ROS in AM was decreased, and the level of Ca 2+ in AM was increased in test group ( P<0.05). Conclusions:The mechanism by which implementing AOI in the non-ventilated lung reduces lung injury may be related to promotion of transformation of AM from M1 phenotype to M2 phenotype and inhibition of inflammatory responses during OLV in the patients undergoing thoracoscopic lobectomy.

Effects of dexmedetomidine on alveolar epithelial barrier function in rats with VILI and the role of PKC / 中华麻醉学杂志

Objective:To evaluate the effects of dexmedetomidine on alveolar epithelial barrier function in rats with ventilator-induced lung injury (VILI), and the role of protein kinase C (PKC).Methods:One hundred clean-grade male Sprague-Dawley rats, weighing 270-320 g, aged 4-5 months, were divided into 5 groups ( n=20 each) using a random number table method: control group (group C), VILI group (group V), PKC inhibitor group (group B), dexmedetomidine group (group D), and dexmedetomidine plus PKC agonist group (DP group). The VILI model was developed by mechanical ventilation with a tidal volume of 40 ml/kg for 4 h in anesthetized animals.Group C breathed air autonomously for 4 h without mechanical ventilation.Group V was mechanically ventilated for 4 h. In group B, bisindolvlmaleimide I 0.12 mg/kg was injected intramuscularly 1 h before mechanical ventilation.In D and DP groups, dxmedetomidine 5.0 μg/kg was injected intravenously at 20 min before mechanical ventilation, and dexmedetomidine was intravenously infused at the rate of 5.0 μg·kg -1·h -1 during mechanical ventilation.In group DP, PKC agonist phorbol-12-myristic acid-13-acetate 15 μg/kg was intraperitoneally injected at 30 min before mechanical ventilation.At 4 h of mechanical ventilation, oxygenation index (OI), lung permeability index (LPI) and wet/dry lung weight (W/D) ratio were measured, the pathological changes of lung tissues were observed, and lung injury was assessed and scored.The expression of PKC, occludin and ZO-1 protein was detected by Western blot, and the expression of PKC mRNA, occludin mRNA and ZO-1 mRNA was determined by real-time polymerase chain reaction. Results:Compared with group C, OI was significantly decreased, LPI, W/D ratio and lung injury score were increased, the expression of PKC protein and mRNA was up-regulated, and the expression of occludin and ZO-1 protein and mRNA was down-regulated in V and DP groups ( P<0.05), and no significant change was found in the parameters mentioned above in B and D groups ( P>0.05). Compared with group V, OI was significantly increased, LPI, W/D ratio and lung injury score were decreased, the expression of PKC protein and mRNA was down-regulated, and the expression of occludin and ZO-1 protein and mRNA was up-regulated in B, D and DP groups ( P<0.05). Compared with group D, OI was significantly decreased, LPI, W/D ratio and lung injury score were increased, the expression of PKC protein and mRNA was up-regulated, and the expression of occludin and ZO-1 protein and mRNA was down-regulated in group DP ( P<0.05). Conclusions:Dexmedetomidine can reduce the damage to alveolar epithelial barrier function in rats with VILI, and the mechanism is related to inhibition of PKC activation and up-regulation of the expression of occludin and ZO-1.

Relationship between HO-1 and PPARγ during alveolar macrophage polarization in a mouse model of endotoxin-induced acute lung injury / 中华麻醉学杂志

Objective:To evaluate the relationship between heme oxygenase-1 (HO-1) and peroxisome proliferator-activated receptor γ (PPARγ) during alveolar macrophage polarization in a mouse model of endotoxin-induced acute lung injury (ALI).Methods:Thirty clean-grade male C57BL/6 mice (24 wide-type mice and 6 HO-1 knockout mice), aged 6-8 weeks, weighing 18-22 g, were studied.Wide-type mice were divided into 4 groups ( n=6 each) using a random number table method: control group (C group), ALI group, ALI+ HO-1 agonist hemin group (ALI+ H group), and ALI+ hemin+ PPARγ antagonist T0070907 group (ALI+ H+ T group).HO-1 knockout mice in which the ALI model was developed served as ALI+ HO-1 -/- group.ALI model was developed by injecting lipopolysaccharide (LPS) 15 mg/kg via the tail vein in anesthetized animals.T0070907 1.5 mg/kg was intraperitoneally injected at 1 h before LPS administration in ALI+ H+ T group, and hemin 50 mg/kg was intraperitoneally injected at 30 min before LPS administration in ALI+ H group and ALI+ H+ T group.Mice were sacrificed at 12 h after LPS administration, and lung tissues were obtained to measure the wet to dry weight ratio (W/D ratio), to observe pathological changes which were scored, and to determine the F4/80+ /CD86+ labeled M1 alveolar macrophages and the F4/80+ /CD206+ labeled M2 alveolar macrophages (by flow cytometry), contents of M1 macrophage-related genes inducible nitric oxide synthase (iNOS) and M2 macrophage-related genes Arginase-1 (Arg-1) (by enzyme-linked immunosorbent assay), and the expression of HO-1 and PPARγ (by Western blot). Results:Compared with C group, the lung injury score, W/D ratio, levels of CD86 and CD206, and contents of iNOS and Arg-1 were significantly increased, and PPARγ expression was up-regulated in the other four groups ( P<0.05), and HO-1 protein expression was up-regulated in ALI, ALI+ H and ALI+ H+ T groups ( P<0.05).Compared with ALI group, the lung injury score, W/D ratio, and levels of CD86 and iNOS were significantly increased, the levels of CD206 and Arg-1 were decreased, and the expression of HO-1 and PPARγ was down-regulated in ALI+ HO-1 -/- group, the lung injury score, W/D ratio and levels of CD86 and iNOS were significantly decreased, the levels of CD206 and Arg-1 were increased, and the expression of HO-1 and PPARγ was up-regulated in ALI+ H group ( P<0.05), and no significant change was found in the parameters mentioned above in ALI+ H+ T group ( P>0.05).Compared with ALI+ H group, the lung injury score, W/D ratio and levels of CD86 and iNOS were significantly increased, the levels of CD206 and Arg-1 were decreased, the expression of PPARγ was down-regulated ( P<0.05), and no significant change was found in the expression of HO-1 in ALI+ H+ T group ( P>0.05). Conclusions:HO-1 can up-regulate the expression of PPARγ, inhibit the polarization of alveolar macrophages toward M1 phenotype and promote the polarization toward M2 phenotype, thus playing an endogenous protective role in endotoxin-induced ALI in mice.

Lung Ultrasound as a Triage Tool in an Emergency Setting during the Covid-19 Outbreak: comparison with CT Findings

Abstract Background Lung ultrasound (LUS) can detect interstitial alveolar changes confined to the subpleural region, like those described in Covid-19. Objetive To evaluate how LUS findings correlate with chest computed tomography (CT) in patients admitted to the emergency department (ED) with suspicion of Covid-19. Methods Cross-sectional study of 20 patients (median age 43 years; interquartile range, 37-63 years; 50% male). All patients underwent LUS and chest CT on the day of ED admission. Each hemithorax was divided into 6 segments with similar landmarks, and equivalent scores (sc) of lesion severity were defined for both methods. The number of affected segments on LUS (LUSseg) was divided into tertiles (0-1, 2-5, and ≥6), and compared with number of affected segments on CT (CTseg), LUSsc, CTsc, and percentage of affected lung parenchyma through visual analysis (CTvis). ANOVA or Kruskal-Wallis test for continuous variables, chi-square test for categorical variables, and receiver operating characteristic (ROC) curve analysis to define optimal cutoff points were performed. P<0.05 was considered statistically significant. Results Median LUSsc, CTsc, CTseg, and CTvis were significantly different between groups. A clear separation between groups was demonstrated; patients with <2 affected segments on LUS were defined as low risk. The ROC curve showed good discriminative power to predict ≥6 affected segments on CT, with an area under the curve (AUC) of 0.97 and 0.98 for >7 LUSsc and >3 LUSseg, respectively. Conclusion LUS findings correlate with chest CT, and can help identify patients with normal lung or minor pulmonary involvement secondary to Covid-19. Int J Cardiovasc Sci. 2020; [online].ahead print, PP.0-0

Histological Study of Mammary Lobule on Pregnant, Lactation and Involution Stage in Murrah Buffalo

Present experiment was conducted on sixty Murrah buffalo divided into three groups : pregnant, lactating and involution stage/Dry stage. No distinct lobulation was observed during nonlactating early pregnant stage. The ratio between maximum diameter of lobule in nonlactating mid and late pregnant stage was 4:5. Mammary lobules were oval to polygonal in shape. In lactating stage the approximate ratio between maximum diameter of lobule in colostrum stage and three months of lactation were 2:1. The number of alveoli were maximum during colostrum phase and reduced from colostrum stage to ten months of lactation. Highly significant statistical difference in the diameter of lobules and number of alveoli was noticed during different stages of lactation from colostrum to ten month. The number of alveoli was minimum during nonlactating nonpregnant stage from one to two month

Hemorragia alveolar difusa no inmune en pediatría / Non-inmune mediated diffuse alveolar hemorrage in children / A hemorragia alveolar difusa não imune em pediatria

Resumen: La hemorragia alveolar difusa no inmune es una patología poco frecuente tanto en la población pediátrica como en los adultos. Si bien se conocen factores determinantes de dicha patología, la interacción de éstos a la hora de producirse el sangrado no es del todo comprendida. Para cumplir con la función de intercambio gaseoso la membrana alvéolo-capilar debe tener un espesor bajo, lo que la expone a falla por estrés y eventualmente extravasación de sangre al parénquima pulmonar. La membrana basal, sumado a la disposición de los alvéolos y el intersticio pulmonar, le dan resistencia extraordinaria a dicha membrana. Si ocurre efracción de la membrana alvéolo capilar puede haber pasaje de sangre al alvéolo e intersticio pulmonar, este fenómeno es conocido como falla por estés. En los casos que existe fragilidad de la membrana alvéolo-capilar, ya sea por inmadurez o por situaciones patológicas, existe un riesgo significativo de efracción de la misma. Alteraciones hemodinámicas y de la cascada de la coagulación pueden ser factores determinantes en el desarrollo de hemorragia alveolar. En el presente trabajo se discuten cuatro casos clínicos de hemorragia alveolar difusa no inmune y en base a éstos se desarrolla una hipótesis con el objetivo de explicar la interrelación de los factores que inciden en el desarrollo de dicha entidad.

Summary: Non-immune mediated alveolar hemorrhage is a rare pathology affecting both children and adults. Although the causes of this disease are known, their interaction in case of bleeding is not clearly understood. The present paper discusses 4 clinical cases of non-immune mediated alveolar hemorrhage and a hypothesis is presented with the objective of explaining the interrelation of the factors that may affect the its development. In order to comply with the gas exchange function, the alveolar-capillary membrane must not be too thick, since this might lead to stress failure and to eventual blood extravasation to the lung parenchyma. The basement membrane (type IV) collagen, plus the alveoli arrangement and the pulmonary interstitium, provide extraordinary resistance to such membrane. Whenever the alveolar-capillary membrane is weak, either due to immaturity or to pathological situations, there is a significant risk of rupture. Hemodynamic and coagulation alterations can be determining factors in the development of this pathology, since they affect the development of stress failure or perpetuate the passage of blood to the pulmonary interstitium.

Resumo: A hemorragia alveolar não imune é uma patologia rara tanto nas crianças como nos adultos. Embora os determinantes dessa patologia sejam conhecidos, sua interação no momento de determinar o sangramento não é completamente compreendida. Neste estudo discutimos quatro casos clínicos de hemorragia alveolar não imune e desenvolvimos hipóteses para explicar a inter-relação de fatores que afetam o desenvolvimento da doença. Para que a membrana possa cumprir a sua função de troca de gás alveolar-capilar deve ter baixa espessura, o que a expõe à tensão de ruptura e, eventualmente, o extravasamento de sangue para o parênquima pulmonar. O colagénio tipo IV da membrana basal, adicionado à disposição dos alvéolos e ao interstício pulmonar, confere extraordinária resistência à referida membrana. Nos casos nos que há fragilidade da membrana alvéolo-capilar, seja por imaturidade ou por situações patológicas, existe risco significativo de sua ruptura. Alterações hemodinâmicas e de coagulação podem ser fatores determinantes no desenvolvimento dessa patologia, uma vez que afetam o desenvolvimento de falha de estresse ou perpetuam a passagem do sangue para o interstício pulmonar.

Histogenesis Of Human Lung - Retrospective Fetal Autopsy Study

Histogenesis of human fetal lung was studied in 40 human fetuses under light microscope after sectioning andstaining with hematoxylin & eosin stain, in fetuses with gestational age ranging from 10 weeks to fetuses above30 weeks. Appearance of various levels of bronchi were identified with the changes in the epithelium at differentlevels of bronchi. The appearance of alveolar ducts and few alveoli were recognized after 25 weeks of gestation.Vascularisation of the fetal lung was observed as early as 10 weeks of gestation. Appearance of various levels ofbronchi were identified with the changes in the epithelium at different ages of gestation

Endotoxin-induced endogenous protective mechanism of alveolar type Ⅱ epithelial cells of rats: the relationship with p38MAPK-HO-1-mitochondrial fusion signaling pathway / 中华麻醉学杂志

Objective To evaluate the endotoxin-induced endogenous protective mechanism of alveolar type Ⅱ epithelial cells of rats and the relationship with p38 mitogen-activated protein kinase (p38MAPK)-HO-1-mitochondrial fusion signaling pathway.Methods Rat alveolar type Ⅱ epithelial cells were seeded in 6-well plates at a density of 2× 105 cells/ml and divided into 5 groups (n =15 each) using a random number table method:control group (group C),lipopolysaccharide (LPS) group (group L),LPS plus p38MAPK inhibitor SB203580 group (group LS),LPS plus dimethyl sulfoxide group (group LD),and SB203580 group (group S).Cells were conventionally cultured in group C.The model of endotoxin-challenged alveolar type Ⅱ epithelial cells was established by giving LPS 10 μg/ml in L,LS and LD groups.SB203580 10 μmol and 0.1％ dimethyl sulfoxide 100 μμmol were added at 1 h before giving LPS in group LS and group LD,respectively.SB203580 10 μ mol was added to the culture medium in group S.All the cells were incubated for 24 h.The malonaldehyde (MDA) content and superoxide dismutase (SOD) activity in the culture medium were determined by thiobarbituric acid assay and xanthine oxidase method,respectively.The expression of p38MAPK,phosphorylated p38MAPK (p-p38MAPK),hemeoxygenase-1 (HO-1),mitofusin 1 (Mfn1),Mfn2,and optical atrophy-1 (OPA1) was measured by Western blot.Results Compared with group C,the MDA content was significantly increased,the SOD activity was decreased,and the expression of p-p38MAPK and HO-1 was up-regulated,and the expression of Mfn1,Mfn2 and OPA1 was down-regulated in L,LS and LD groups (P＜0.05).Compared with group L,the MDA content was significantly increased,the SOD activity was decreased,and the expression of pp38MAPK,HO-1,Mfn1,Mfn2 and OPA1 was down-regulated in group LS (P＜0.05),and no significant change was found in the indices mentioned above in group LD (P＞0.05).Conclusion The endotoxin-induced endogenous protective mechanism of alveolar type Ⅱ epithelial cells is related to p38MAPK-HO-1-mitochondrial fusion signaling pathway in rats.

Role of LC3Ⅱ in hyperoxia-induced apoptosis in alveolar typeⅡ epithelial cells / 中华麻醉学杂志

Objective@#To investigate the role of microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ) in hyperoxia-induced apoptosis in alveolar typeⅡ epithelial cells.@*Methods@#A549 cells were seeded in 6-well culture plates at a density of 1×105 cells/ml (0.5 ml/well) and were divided into 3 groups (n=12 each) using a random number table method: control group (group C), 95% oxygen group (group 95% O2) and 95% oxygen plus autophagy inhibitor 3-methyladenine (3-MA) group (group 95% O2+ 3-MA). Cells in group C were cultured in incubators containing 95% air and 5% CO2 at 37 ℃.Cells were cultured in sealed chambers flushed with 95% O2 and 5% CO2 at 37 ℃ in group 95% O2 and group 95% O2+ 3-MA.The cells in each well were pretreated with 3-MA 5 mmol/L before exposure to 95% O2 in group 95% O2+ 3-MA.When the A549 cells were incubated for 24 h, 6 wells in each group were selected to detect the expression of LC3Ⅱ protein and caspase-3 using Western blot, and the left 6 wells in each group were selected to calculate the cell mortality rate by trypan blue dye.@*Results@#Compared with group C, the expression of LC3Ⅱ and caspase-3 was significantly up-regulated, and the cell mortality rate was increased in group 95% O2 and group 95% O2+ 3-MA (P<0.05). Compared with group 95% O2, the expression of LC3Ⅱ was significantly down-regulated, the expression of caspase-3 was up-regulated, and the cell mortality rate was increased in group 95% O2 and group 95% O2+ 3-MA (P<0.05).@*Conclusion@#The up-regulated expression of LC3Ⅱ can inhibit hyperoxia-induced apoptosis in alveolar typeⅡ epithelial cells.

Role of LC3 Ⅱ in hyperoxia-induced apoptosis in alveolar type Ⅱ epithelial cells / 中华麻醉学杂志

Objective To investigate the role of microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ) in hyperoxia-induced apoptosis in alveolar type Ⅱ epithelial cells.Methods A549 cells were seeded in 6-well culture plates at a density of 1 × 105 cells/ml (0.5 ml/well) and were divided into 3 groups (n =12 each) using a random number table method:control group (group C),95％ oxygen group (group 95％ O2) and 95％ oxygen plus autophagy inhibitor 3-methyladenine (3-MA) group (group 95％ O2+3-MA).Cells in group C were cultured in incubators containing 95％ air and 5％ CO2 at 37 ℃.Cells were cultured in sealed chambers flushed with 95％ O2 and 5％ CO2 at 37 ℃ in group 95％ O2 and group 95％ O2+3-MA.The cells in each well were pretreated with 3-MA 5 mmol/L before exposure to 95％ O2 in group 95％ O2+3-MA.When the A549 cells were incubated for 24 h,6 wells in each group were selected to detect the expression of LC3 Ⅱ protein and caspase-3 using Western blot,and the left 6 wells in each group were selected to calculate the cell mortality rate by trypan blue dye.Results Compared with group C,the expression of LC3 Ⅱ and caspase-3 was significantly up-regulated,and the cell mortality rate was increased in group 95％ O2 and group 95％ O2+3-MA (P＜0.05).Compared with group 95％ O2,the expression of LC3 Ⅱ was significantly down-regulated,the expression of caspase-3 was up-regulated,and the cell mortality rate was increased in group 95％ O2 and group 95％ O2+3-MA (P＜0.05).Conclusion The up-regulated expression of LC3 Ⅱ can inhibit hyperoxia-induced apoptosis in alveolar type Ⅱ epithelial cells.

Poliangiite microscópica com síndrome pulmão-rim: relato de caso e revisão de literatura / Microscopic polyangiitis with lung-kidney syndrome: case report and literature review

A poliangiite microscópica (PAM) é uma vasculite necrosante sistêmica pauci-imune associada ao anticorpo anticitoplasma de neutrófilos (ANCA) com preferência de pequenos vasos. Relato do caso: Relatamos o caso de uma paciente do sexo feminino, 54 anos, que apresentou quadro de poliartrite migratória em punhos, joelhos e tornozelos associada à rigidez matinal progressiva, com histórico de "rash" malar, fotossensibilidade e alopecia. Progrediu ao longo do ano de 2017 com deterioração da função renal e hemorragia pulmonar, necessitando de cuidados intensivos. A biópsia renal sugeriu padrão compatível com glomerulonefrite pauci-imune e o diagnóstico de poliangiite microscópica foi aventado. Realizou pulsoterapia com metilprednisolona e ciclofosfamida, além de plasmaférese, recebendo alta após estabilização do quadro clínico. Importância do problema: O presente caso ilustra uma complicação incomum e de elevada morbimortalidade da PAM. A negatividade do ANCA dificultou o diagnóstico, sendo necessária a realização de biópsia renal com confirmação diagnóstica. A síndrome pulmão-rim apresenta evolução potencialmente fatal se não instituído precocemente o tratamento. (AU)

Microscopic polyangiitis (MPA) is a pauci-immune systemic necrotizing vasculitis associated with neutrophil anti-cytoplasmic antibody (ANCA) with a preference for small vessels. Case report: We report the case of a 54-year-old woman, who presented migratory polyarthritis in wrists, knees and ankles associated with progressive morning stiffness, with history of malar "rash", photosensitivity and alopecia. It progressed throughout the year of 2017 with deterioration of renal function and pulmonary hemorrhage, requiring intensive care. Renal biopsy suggested a pattern compatible with pauci-immune glomerulonephritis and the diagnosis of microscopic polyangiitis was suggested. She underwent pulse therapy with methylprednisolone and cyclophosphamide, in addition to plasmapheresis, being discharged from hospital after stabilization of the clinical condition. Importance of the issue: The present case reveals an uncommon and high morbimortality complication of MPA. The negativity of the ANCA made diagnosis difficult, and a renal biopsy was necessary to confirm diagnosis. Lung-kidney syndrome is potentially fatal if the treatment is not instituted early. (AU)

Role of protein kinase Cα/heme oxygenase-1 signaling pathway in LPS-caused damage to type Ⅱ alveolar epithelial cells of rats: the relationship with mitochondrial fusion / 中华麻醉学杂志

Objective To evaluate the role of protein kinase Cα (PKCoα)/heme oxygenase-1 (HO-1) signaling pathway in lipopolysaccharide (LPS)-caused damage to type Ⅱ alveolar epithelial cells of rats and the relationship with mitochondrial fusion.Methods Type Ⅱ alveolar epithelial cells were seeded in 96-well plates at a density of 2× 105 cells/ml and divided into 5 groups (n =40 each) using a random number table method:control group (group C),Go6976 group (group G),LPS group (group L),LPS plus PKCα inhibitor Go6976 group (group LG) and LPS plus dimethyl sulfoxide (DMSO) group (group LD).Group LG and group LD were pretreated with 5 μmol/L Go6976 and the equal volume of 0.1％ DMSO,respectively,for 30 min,lipopolysaccharide (LPS) 10 μg/ml was then given to establish the model of type Ⅱ alveolar epithelial cell damage in L,LG and LD groups,Go6976 5 μmol/L was added in group G,and the equal volume of phosphate buffer solution was added in group C.The cells were collected after 24 h of incubation for measurement of malondialdehyde (MDA) and reactive oxygen species (ROS) contents,superoxide dismutase (SOD) activity and expression of PKCα,HO-1,mitochondrial fusion-related proteins 1 and 2 (Mfn1,Mfn2),optic atrophy 1 (OPA1) protein and mRNA (by fluorescent quantitative polymerase chain reaction or Western blot).Results Compared with group C,MDA and ROS contents were significantly increased,the SOD activity was decreased,the expression of PKCα and HO-1 protein and mRNA was up-regulated,and the expression of Mfn1,Mfn2 and OPA1 protein and mRNA was downregulated in L,LG and LD groups (P＜0.05),and no significant change was found in the parameters mentioned above in group G (P＞0.05).Compared with group L,MDA and ROS contents were significantly increased,the SOD activity was decreased,and the expression of PKCα,HO-1,Mfn1,Mfn2 and OPA1 protein and mRNA was down-regulated in group LG (P＜0.05),and no significant change was found in the parameters mentioned above in group LD (P＞0.05).Conclusion Activation of PKCα/HO-1 signaling pathway is the endogenous protective mechanism of LPS-caused damage to type Ⅱ alveolar epithelial cells,which may be related to promoting mitochondrial fusion in rats.

Experimental research on the effect of functional residual capacity on the deposition of inhalable particles in human alveoli region / 生物医学工程学杂志

Research on the deposition of inhalable particles in the alveoli of the lungs is important to the causes, development for common respiratory diseases such as emphysema, and even the optimization of clinical treatment and prevention programs of them. In this paper, an experimental model was established to simulate the deposition of terminal bronchioles and pulmonary acinus particles. The deposition rate of inhalable particles with different particle sizes in the pulmonary acinus was studied under different functional residual capacity. The results showed that the particle diameter was an important factor affecting the deposition of particles in the lung alveoli. Particles with 1 μm diameter had the highest deposition rate. With the functional residual capacity increasing, particulate deposition rate significantly reduced. The results of this study may provide data support and optimization strategy for target inhalation therapy of respiratory diseases such as emphysema and pneumoconiosis. The established model may also provide a feasible experimental model for studying the deposition of inhalable particles in the pulmonary alveoli.

Role of PI3K/Akt signaling pathway in carbon monoxide-induced up-regulation of mitochondrial fusion proteins in endotoxin-challenged alveolar epithelial cells of rats / 中华麻醉学杂志

Objective To evaluate the role of 1-phosphatidylinositol 3-kinase/serine threonine kinase (PI3K/Akt) signaling pathway in carbon monoxide (CO)-induced up-regulation of mitochondrial fusion proteins in endotoxin-challenged alveolar epithelial cells of rats.Methods The rat alveolar epithelial cells were cultured in a 5％ CO2 cell culture incubator at 37 ℃ with F12K complete medium containing 10％ fetal bovine serum and 1％ green chain double antibody,and divided into 10 groups (n=5 each) according to the random number table method:control group (C group),endotoxin group (L group),lipopolysaccharide (LPS) plus exogenous CO-releasing agent CO-releasing-molecule-2 (CORM-2) group (L+CO group),LPS plus PI3K inhibitor LY294002 group (L+LY group),LPS plus CORM-2 plus LY294002 group (L+ CO+LY group),LPS plus inactive CO releasing agent iCORM-2 group (L+iCO group),LPS plus dimethyl sulfoxide (DMSO) group (L+D group),CORM-2 group (CO group),LY294002 group (LY group) and CORM-2 plus LY294002 group (CO+LY group).LPS 10 μg/ml was added to the culture medium in group L.CORM-2 100 μmol/L was added to the culture medium and 1 h later 10 μg/ml LPS was added in L+CO group.LY294002 25 μmol/L was added to the medium,and 1 h later LPS 10 μg/ml was added in L+LY group.In L+CO+LY group,25 μmol/L LY294002 was added to the culture medium,CORM-2 100 μmol/L was added 1 h later,and then LPS 10 μg/ml was added 1 h later.iCORM 100 μmol/L was added to the culture medium and 1 h later LPS 10 μg/ml was added in L+iCO group.In L+D group,the equal concentration of DMSO was added to the culture medium and 1 h later LPS 1 μg/ml was added.CORM-2 100 μmol/L was added to the culture medium in CO group.LY294002 25 μmol/L was added to culture medium in LY group.LY294002 25 μmol/L was added to the culture medium,and 1 h later CORM-2 100 μmol/L was added in CO+LY group.Cells were harvested after 24 h of incubation for measurement of the malondialdehyde (MDA) content,superoxide dismutase (SOD) activity and expression of heme oxygenase-1 (HO-1),phosphorylated Akt (p-Akt),mitochondrial fusion proteins mitofusion 1 (Mfn1),Mfn2 and optic atrophy 1 (OPA1) by Western blot.Results Compared with group C,the MDA content was significantly increased and SOD activity was decreased in L,L+CO,L+LY,L+CO+LY,L+iCO,L+D groups,and the expression of HO-1,Mfn1,Mfn2,OPA1 protein and p-Akt was significantly up-regulated in group L (P＜0.05).Compared with group L,MDA content was significantly decreased and SOD activity was increased in group L+CO,MDA content was significantly increased and SOD activity was decreased in group L+LY,the expression of HO-1,Mfn1,Mfn2,OPA1 protein and p-Akt was significantly up-regulated in group L+ CO,and the expression of HO-1,Mfn1,Mfn2,OPA1 protein and p-Akt was significantly down-regulated in group L+LY (P＜0.05).Compared with group L+CO,the MDA content was significantly increased,SOD activity was decreased,and the expression of HO-1,Mfn1,Mfn2,OPA1 and p-Akt was down-regulated in group L+CO+LY (P＜0.05).Conclusion The mechanism by which CO up-regulates the expression of mitochondrial fusion proteins in endotoxin-challenged alveolar epithelial cells is related to activating PI3K/Akt signaling pathway in rats.

Role of PI3K/Akt signaling pathway in mitochondrial fission in endotoxin-challenged alveolar type Ⅱ epithelial cells of rats / 中华麻醉学杂志

Objective To evaluate the role of phosphatidylinositol 3-kinase/serine-threonine kinase(PI3K/Akt) signaling pathway in mitochondrial fission in endotoxin-challenged alveolar type Ⅱ epithelial cells of rats.Methods Rat alveolar type Ⅱ epithelial cells CCL-149 were seeded in 6-well plates at a density of 2×105 cells/ml.CCL-149 cells were divided into 6 groups (n =10 each) using a random number table:control group (group C),lipopolysaccharide (LPS) group (group L),LPS plus CO-releasing molecule-2 (CORM-2) group (group L+CO),LPS plus PI3K inhibitor LY294002 group (group L+LY),LPS plus iCORM-2 group (group L+iCO) and LPS plus dimethyl sulfoxide (DMSO) group (group L+D).CCL149 cells were stimulated with 10 μg/ml LPS for 24 h in L,L+CO,L+LY,L+iCO and L+D groups.CORM-2 100 μmol,LY294002 25 μg and iCORM-2 100 μmol were added at 1 h before stimulation with LPS in L+CO,L+LY and LPS+iC0 groups,respectively.In group L+D,0.1％ DMSO 100 μmol was added at 1 h before stimulation with LPS.After the end of incubation,the concentrations of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the culture medium were determined by enzyme-linked immunosorbent assay,and the expression of phosphorylated-Akt (p-Akt),heme oxygenase-1 (HO-1),dynamin-related protein 1 (Drpl) and fissionl (Fisl) was detected by Western blot.Results Compared with group C,IL-6 and TNF-α concentrations in the culture medium were significantly increased,and the expression ofp-Akt,HO-1,Drp1and FIS1 was up-regulated in L,L+CO,L+LY,L+iCO and L+D groups (P＜0.05).Compared with group L,IL-6 and TNF-α concentrations in the culture medium were significantly decreased,the expression of p-Akt and HO-1 was up-regulated,and the expression of Drp1 and Fis1 was down-regulated in group L+CO,and IL-6 and TNF-α concentrations in the culture medium were significantly increased,the expression of p-Akt and HO-l was down-regulated,and the expression of Drpl and Fisl was up-regulated in group L+LY (P＜0.05).Conclusion Activation of PI3K/Akt signaling pathway can inhibit mitochondrial fission in endotoxin-challenged alveolar type Ⅱ epithelial cells of rats.

Regeneration of functional alveoli by adult human SOX9 airway basal cell transplantation

Irreversible destruction of bronchi and alveoli can lead to multiple incurable lung diseases. Identifying lung stem/progenitor cells with regenerative capacity and utilizing them to reconstruct functional tissue is one of the biggest hopes to reverse the damage and cure such diseases. Here we showed that a rare population of SOX9 basal cells (BCs) located at airway epithelium rugae can regenerate adult human lung. Human SOX9 BCs can be readily isolated by bronchoscopic brushing and indefinitely expanded in feeder-free condition. Expanded human SOX9 BCs can give rise to alveolar and bronchiolar epithelium after being transplanted into injured mouse lung, with air-blood exchange system reconstructed and recipient's lung function improved. Manipulation of lung microenvironment with Pirfenidone to suppress TGF-β signaling could further boost the transplantation efficiency. Moreover, we conducted the first autologous SOX9 BCs transplantation clinical trial in two bronchiectasis patients. Lung tissue repair and pulmonary function enhancement was observed in patients 3-12 months after cell transplantation. Altogether our current work indicated that functional adult human lung structure can be reconstituted by orthotopic transplantation of tissue-specific stem/progenitor cells, which could be translated into a mature regenerative therapeutic strategy in near future.

Hemosiderosis pulmonar idiopática en paciente adulto / Idiopathic pulmonary hemosiderosis in an adult patient

Resumen Se presenta el caso de una paciente de 46 años de edad con cinco años de evolución de episodios intermitentes de hemoptisis cuyo diagnóstico final fue hemosiderosis pulmonar idiopática. Su presentación y características clínicas son comparadas con los otros casos reportados en la literatura.

Abstract The case of a 46-year-old patient with five years of evolution of intermittent episodes of hemoptysis whose final diagnosis was idiopathic pulmonary hemosiderosis is reported. Its presentation and clinical characteristics are compared with the other cases reported in the literature.

Effects of lipoxin A4 on human type Ⅱ alveolar epithelial cell wound repair, proliferation and apoptosis / 中华麻醉学杂志

Objective To evaluate the effects of lipoxin A4 (LXA4) on human type Ⅱ alveolar epithelial cell wound repair,proliferation and apoptosis.Methods Experiment Ⅰ Human type Ⅱ alveolar epithelial cells were inoculated in 24-well plates and divided into 4 groups (n=10 each) using a random number table:control group (group C),1 nmol/L LXA4 group (group L1),10 nmol/L LXA4 group (group L2) and 100 nmol/L LXA4 group (group L3).Cells were cultured in normal culture atmosphere in group C.Cells were incubated with 1,10 and 100 nmol/L LXA4 in L1,L2 and L3 groups,respectively.The scratch wound assay was performed at 36 h of culture or incubation.Cell proliferation was measured at 24 h of culture or incubation.Experiment Ⅱ Human type Ⅱ alveolar epithelial cells were inoculated in 96-well plates and divided into 5 groups using a random number table:control group (group C,n=10),Fas-ligand group (n =10),Fas-ligand+LXA4 group (n =10),Fas-ligand+TNF-α group (n =5) and Fas-ligand+TNF-α+LXA4 group (n=5).Cells were incubated with 100 ng/ml Fas-Ligand,100 ng/ml Fas-Ligand plus 100 nmol/L LXA4,100 ng/ml Fas-Ligand plus 100 ng/ml TNF-α,and 100 ng/ml Fas-Ligand plus 100 ng/ml TNF-α plus 100 nmol/L LXA4 in Fas-ligand,Fas-ligand+LXA4,Fas-ligand+TNF-α,and Fas-ligand +TNF-α+LXA4 groups,respectively.The cell viability was measured at 24 h of culture or incubation.Cell apoptosis was detected using the flow cytometry,and apoptosis rate was calculated in C,Fas-ligand and Fas-ligand+LXA4 groups.Results Experiment Ⅰ Compared with group C,the percentage of cell repair size and percentage of proliferation were significantly increased in L1,L2 and L3 groups (P＜0.05 or 0.01).Compared with group L1,the percentage of cell repair size and percentage of proliferation were significantly increased in group L3 (P＜ 0.01),and no significant change was found in the parameters mentioned above in group L2 (P＞0.05).Experiment Ⅱ Compared with group C,the cell viability was significantly decreased,and the apoptosis rate was increased in group Fas-ligand,the cell viability was significantly decreased in group Fas-ligand+TNF-α (P＜ 0.01),and no significant change was found in the cell viability or apoptosis rate in group Fas-ligand+LXA4 or in the cell viability in group Fas-ligand+TNF-α+LXA4 (P＞0.05).Compared with group Fas-ligand,the cell viability was significantly increased,and the apoptosis rate was decreased in group Fas-ligand+LXA4 (P＜ 0.05).The cell viability was significantly higher in group Fas-ligand +TNF-α + LXA4 than in group Fas-ligand +TNF-α (P ＜ 0.01).Conclusion LXA4 can promote human type Ⅱ alveolar epithelial cell wound repair and proliferation and inhibit the apoptosis.